studies

208A AASLD ABSTRACTS HEPATOLOGY, October, 2015 

1 

Two Combined Genetic Markers Identify Hepatitis C 

Recipients With a Lower Graft Survival From The Earliest 

Post-Transplant Period 

Silvia Martini 1 , Renato Romagnoli 2 , Francesco Tandoi 2 , Dominic 

Dell’Olio 3 , Paola Magistroni 3,4 , Francesca E. Bertinetto 3,4 , Ennia 

Dametto 3,4 , Stefano Mirabella 2 , Giorgia Rizza 2 , Donatella Cocchis 

2 , Antonio Ottobrelli 1 , Mario Rizzetto 1 , Mauro Salizzoni 2 , 

Antonio Amoroso 3,4 ; 1 Gastrohepatology - Molinette Hospital, 

Turin, Italy; 2 Surgery, Liver Transplant Center - Molinette Hospital, 

Turin, Italy; 3 Regional Transplantation Center, Piedmont, Turin, 

Italy; 4 Immunogenetics Laboratory - Molinette Hospital, Turin, Italy 

Background and Aims. Human Leukocyte Antigen (HLA) variants 

and Interleukin 28B (IL28B) gene are associated with 

recurrence and progression of hepatitis C virus (HCV) liver 

disease in the transplant setting. We investigated the impact 

of these immunogenetic factors on graft survival after liver 

transplantation (LT) for HCV-related cirrhosis. Methods. This 

retrospective study consecutively included all the first adult LTs 

performed at a single center from 1999 to 2009. HLA-A/B/ 

DRB1 frequencies were studied in 1,228 adult Caucasian LT 

recipients and their deceased donors. IL28B rs12979860 

polymorphism was determined only in the 449 HCV-positive 

viremic recipients and their HCV-negative donors, as it does 

not affect transplant outcome in non-HCV patients. Graft survival 

was the sole endpoint. Results. Median follow-up was 10 

years. HLA-DRB1*11 phenotypic frequency was significantly 

lower in HCV-positive recipients compared with HCV-negative 

recipients (28.2% vs 43.9%, p

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AASLD ABSTRACTS 209A 

3 

Determinants and impact of early viral kinetics in 

patients with HCV-recurrence after liver transplantation 

treated by sofosbuvir + daclatasvir: results from the 

ANRS CO23 CUPILT study 

Camille Herve 1 , Audrey Coilly 2 , Claire Fougerou-Leurent 3 , Victor 

de Ledinghen 4 , Georges-Philippe Pageaux 5 , Pauline Houssel-Debry 

3 , Sylvie Radenne 6 , Christophe Duvoux 7 , Vincent Di 

Martino 8 , Nassim Kamar 9 , Louis d’Alteroche 10 , Valerie Canva 11 , 

Pascal Lebray 12 , Jérôme Dumortier 13 , Christine Silvain 14 , Camille 

Besch 15 , Danielle Botta-Fridlund 16 , Albert Tran 17 , Helene Montialoux 

18 , Emilie Rossignol 3 , Alexandra Rohel 19 , Jean-Charles 

Duclos-Vallee 2 , Vincent Leroy 1 ; 1 Gastroenterology, CHU Grenoble, 

La Tronche, France; 2 Paul Brousse, Villejuif, France; 3 CHU 

Rennes, Rennes, France; 4 CHU Bordeaux, Pessac, France; 5 CHU 

Bordeaux, Bordeaux, France; 6 Croix Rousse, Lyon, France; 7 Henri 

Mondor, Créteil, France; 8 CHU Besançon, Besançon, France; 

9 CHU Toulouse, Toulouse, France; 10 CHU Tours, Tours, France; 

11 CHRU Lille, Lille, France; 12 La Pitié Salpétrière, Paris, France; 

13 Hôpital Edouard Herriot, Lyon, France; 14 CHU Poitiers, Poitiers, 

France; 15 CHU Strasbourg, Strasbourg, France; 16 Conception 

Hospital, Marseilles, France; 17 CHU Nice, Nice, France; 18 CHU 

Rouen, Rouen, France; 19 ANRS, Paris, France 

Early viral kinetic does not seem to impact the chance of SVR12 

with the use of new combinations of direct HCV antiviral agents. 

However, few data have been reported in HCV-recurrence after 

liver transplantation (LT). It could be hypothesized that immunosuppression 

might slow down HCV RNA decay. The aims 

of our study were to describe in a large cohort of treated LT 

patients the HCV RNA kinetics, to identify the predictive factors 

of slow response and to evaluate its impact on SVR. Methods: 

One hundred and ninety four LT patients prospectively included 

in the multi-centric CUPILT cohort and treated by sofosbuvir + 

daclatasvir ± ribavirin for 12 or 24 weeks were studied. HCV 

RNA was assessed at W0, W2, W4, W8, W12, EOT and 

FU12 by real time quantitative PCR with a lower limit of quantification 

of 15 IU/ml. Results: The characteristics of patients 

were as follows: age: 58.9 ± 8.7 years, baseline HCV RNA: 

6.4 log IU/ml, genotype 1: 79%, genotype 3: 11%, treatment 

experienced: 47%, HIV co-infection: 7%. The delay between LT 

and onset of treatment was 75.8 ± 69.1 months. The majority 

(60%) had severe HCV recurrence (F3, F4 or FCH). Immunosuppression 

was based on tacrolimus in 62%, cyclosporine in 

28% and everolimus in 8% of cases. MMF was administered 

in 57% of patients. Treatment duration was 24 weeks in 85% 

of cases and 12 weeks in 15% of cases. Ribavirin was used 

in 71%) of patients. The percentages of patients with HCV 

RNA ≥ 15 IU/ml and HCV RNA < 15 IU/ml but detectable 

were distributed as follow: W2: 83% and 13%, W4: 47% and 

29%, W8: 13% and 26%, W12 4% and 14%. By multivariate 

analysis, independent factors associated with HCV RNA ≥ 

15 IU/ml at W4 were: use of MMF OR 1.90 (0.99 – 3.66), 

p3 treatments. 

Intention-to-treat 1- and 5-year overall survival was 84% and 

56%, respectively. Among 109 LT recipients, the 1- and 5-year 

post-LT survival was 95% and 80%, respectively, after a median 

post-LT follow-up of 4.3 years. The Kaplan-Meier probabilities 

of treatment failure at 1 and 5 years from first down-staging 

procedure were 25% and 44%, respectively. Treatment failure 

included dropout due to tumor progression (n=56), liver-related 

death without LT (n=12), and HCC recurrence after LT (n=12). 

In multivariable Cox proportional hazards regression analy-


sis, factors predicting treatment failure were pre-treatment AFP 

>1000 ng/mL (HR 3.3, p1000 ng/mL compared 

to 40% for AFP


and Status 1A FHF patients may be over-prioritized according 

to current allocation policies. 

Disclosures: 

W. Ray Kim - Advisory Committees or Review Panels: Bristol Myers Squibb, 

Gilead Sciences, Abbvie, Merck 

Patrick S. Kamath - Advisory Committees or Review Panels: Sequana Medical 

The following authors have nothing to disclose: Alina M. Allen, Julie Heimbach, 

Joseph J. Larson, Terry M. Therneau 

7 

End Stage Liver Disease Patients with MELD Scores 

Over 40 Have Significantly Higher Waitlist Mortality 

and Lower Probability of Receiving Liver Transplantation 

Compared to Status 1A Fulminant Hepatic Failure 

Patients 

Joseph C. Ahn 2 , Taft Bhuket 1 , Sasan Mosadeghi 1 , Catherine T. Frenette 

3 , Benny Liu 1 , Robert J. Wong 1 ; 1 Gastroenterology and Hepatology, 

Alameda Health System - Highland Hospital, Oakland, CA; 

2 School of Medicine, University of California, San Francisco, San 

Francisco, CA; 3 Organ Transplantation, Scripps Green Hospital, 

San Diego, CA 

Background: Current liver transplantation (LT) allocation systems 

prioritize Status 1A patients with fulminant hepatic failure 

(FHF) over end stage liver disease (ESLD) patients of all MELD 

scores. However, ESLD patients with the highest MELD scores 

may have higher waitlist mortality than Status 1A FHF patients, 

and thereby may require LT more urgently. Aim: Evaluate differences 

in LT waitlist mortality and probability of LT between Status1A 

FHF patients and ESLD patients with MELD>30. Methods: 

Using data from United Network for Organ Sharing registry, 

we retrospectively evaluated U.S. adults (age > 18) who were 

listed for LT from January 1, 2003 to December 31, 2013. 

ESLD patients were stratified into 3 groups of MELD scores (31- 

35, 36-40, > 40). Overall waitlist mortality and probability 

of LT were stratified by ESLD vs. Status 1A FHF and evaluated 

with Kaplan Meier curves and multivariate logistic regression 

models. The final multivariate model included adjustments for 

age, sex, race/ethnicity, obesity, hepatic encephalopathy, 

and ascites. Results: From 2003-2013, 15,049 ESLD patients 

with MELD>30 and 3,049 Status 1A FHF patients were listed 

for LT. ESLD patients with MELD 31-35 and MELD 36-40 had 

significant higher 28-day waitlist survival than Status 1A FHF 

patients, whereas MELD >40 patients had similar 28-day survival. 

Compared to Status 1A FHF patients, overall probability 

of LT was similar among ESLD patients with MELD 36-40 and 

MELD>40. On multivariate regression, compared to Status 1A 

FHF, ESLD patients with MELD scores >40 had significantly 

higher 14-day waitlist mortality (OR 1.92; 95% CI 1.56-2.36; 

p40 have significantly higher waitlist mortality compared 

to Status 1A FHF patients. Despite the higher waitlist mortality, 

ESLD patients with MELD >40 had similar probability of receiving 

LT, suggesting that these patients may be under-prioritized 

Disclosures: 

Catherine T. Frenette - Speaking and Teaching: Bayer, Salix, Gilead; Stock 

Shareholder: Gilead 

The following authors have nothing to disclose: Joseph C. Ahn, Taft Bhuket, 

Sasan Mosadeghi, Benny Liu, Robert J. Wong 

8 

Enhancing hepatocyte function using an advanced 

microfluidic system for clinical and pharmaceutical 

applications 

Maria Navarro-Zornoza 1,2 , Xavi Illa 2 , Carmen Peralta 3 , Rosa 

Villa 2 , Jordi Gracia-Sancho 1 ; 1 Barcelona Hepatic Hemodynamic 

Lab, IDIBAPS - Hospital Clinic de Barcelona - CIBEREHD, Barcelona, 

Spain; 2 CNM-CSIC, Barcelona, Spain; 3 IDIBAPS - CIBEREHD, 

Barcelona, Spain 

Background and Aims: Development of an advanced liveron-a-chip 

to support liver function in patients with acute liver 

failure and for in vitro drug discovery is still challenging. We 

hypothesized that representation of the unique architectural 

and dynamic features of the hepatic sinusoid using a superior 

microfluidic bioreactor system for co-culture of primary liver 

sinusoidal endothelial cells (LSECs) and hepatocytes would 

mimic the in vivo hepatic metabolism significantly better than 

using conventional methods for hepatocytes mono-culture or 

co-cultures. Methods: Hepatocytes and LSECs freshly isolated 

from Wistar rats were co-cultured in a bioreactor recently 

developed by our team. LSECs were grown in the upper area 

of the bioreactor on a 1mm pore size membrane with homogeneous 

and continuous shear stress stimulation (3dyn/cm 2 ); 

whereas hepatocytes were plated in the lower area. After 

48-72h of co-culture, hepatocyte function and phenotype 

were characterized as albumin synthesis (enzymatic method), 

urea production (BUN enzymatic method), CYP3A4 (luminescence 

assay) and HNF4a expression (qPCR), and compared 

to hepatocytes co-cultured with LSECs in the microfluidic system 

but under static conditions or in conventional transwells, 

and with hepatocytes cultured alone. Results: Hepatocytes 

co-cultured in the bioreactor exhibited markedly better phenotype 

than those cultured using conventional methods. Indeed, 

hepatocytes from the bioreactor maintained polygonal shape 

differentiation, had higher albumin and urea production (Albumin: 

67-fold higher vs. transwells; 108-fold vs. mono-cultures; 

Urea: 79-fold superior vs. transwells; 108-fold vs. mono-cultures), 

exhibited markedly higher HNF4a expression (17-fold 

vs. transwells; 12-fold vs. mono-cultures), and superior CYP3A4 

activity (28-fold vs. transwells; 17-fold vs. mono-cultures). All 

differences were p


actions condition). Conclusion: The herein presented bioreactor 

allows co-culture of LSECs and hepatocytes, maintaining and 

enhancing hepatocyte function long-term significantly better 

than using monocultures or conventional co-culture methods. 

Our approach provides an overwhelming insight in the importance 

of the hepatic sinusoid in the development of technology 

for liver support in acute liver failure and for drug discovery. 

Disclosures: 

The following authors have nothing to disclose: Maria Navarro-Zornoza, Xavi 

Illa, Carmen Peralta, Rosa Villa, Jordi Gracia-Sancho 

9 

Prediction of waitlist mortality in patients with portopulmonary 

hypertension (POPH): An analysis of the UNOS 

database 

Hilary M. DuBrock 1,2 , David S. Goldberg 4 , Norman L. Sussman 5 , 

Sonja Bartolome 6 , Zakiyah Kadry 7 , Reena Salgia 8 , Andre M. 

De Wolf 9 , David C. Mulligan 10 , Steven M. Kawut 4 , Michael J. 

Krowka 3 , Richard Channick 2 ; 1 Medicine, Beth Israel Deaconess 

Medical Center, Boston, MA; 2 Medicine, Massachusetts General 

Hospital, Boston, MA; 3 Mayo Clinic, Rochester, MN; 4 University 

of Pennsylvania, Philadelphia, PA; 5 Baylor College of Medicine, 

Houston, TX; 6 UT Southwestern, Dallas, TX; 7 Penn State Hershey 

Medical Center, Hershey, PA; 8 Henry Ford Hospital, Detroit, MI; 

9 Northwester University, Chicago, IL; 10 Yale New Haven Hospital, 

New Haven, CT 

Purpose: The current United Network for Organ Sharing 

(UNOS) system grants Model for End-Stage Liver Disease 

(MELD) exception points to patients with POPH and a mean 

pulmonary artery pressure (mPAP) 25 and pulmonary vascular 

resistance (PVR)>240] was built using a competing risk subdistribution 

hazard model and purposeful selection. Significance 

was defined as p16 years of age) were approved for 

a POPH MELD exception. In patients with true POPH (n=190), 

13(6.8%) died on the waitlist, 4(2.1%) died during transplant, 

27(14.2%) were removed for being too sick, 100(52.6%) 

patients underwent deceased donor liver transplant, 2(1.1%) 

underwent living donor transplant, 9(4.7%) were removed for 

other reasons and 35(18.4%) remained active on the waitlist. 

Initial MELDNa was a significant univariate predictor of waitlist 

mortality (HR 1.121, p


1%. Conclusion. In a national cohort of HCC patients undergoing 

LT, the recently enacted UNOS criteria improved the accuracy 

of radiographic staging but did not reduce false-positive 

HCC diagnoses. Overall utilization of HCC MELD exceptions 

did not meaningfully change due to a shift from automatic T2 

to RRB-approved exceptions. 

Disclosures: 

Jesse M. Civan - Consulting: Merck 

The following authors have nothing to disclose: Barry Schlansky, Willscott E. 

Naugler 

11 

Treatment with Optifast Reduces Hepatic Steatosis and 

Safely Increases Candidacy Rates for Live Donor Liver 

Transplantation 

Adam Doyle 1 , Jayne Dillon 1 , Oyedele Adeyi 2 , Sandra E. Fischer 2 , 

Meaghan MacArthur 1 , Max Marquez 1 , Robert Smith 1 , David 

Grant 1 , Mark S. Cattral 1 , Ian McGilvray 1 , Paul Greig 1 , Anand 

Ghanekar 1 , Markus Selzner 1 , Eberhard L. Renner 1 , Les Lilly 1 , Gary 

Levy 1 , Nazia Selzner 1 ; 1 Multi Organ Transplant Program, University 

Health Network, Toronto, ON, Canada; 2 Department of 

Pathology, University Health Network, Toronto, ON, Canada 

Background: As a consequence of the increased prevalence 

of obesity and associated liver steatosis, the volunteer pool for 

live donor liver transplantation is threatened. The Optifast verylow 

calorie diet meal replacement system has demonstrated 

efficacy in the reduction in obesity and hepatic steatosis in 

candidates for bariatric surgery. We studied the ability of 

a 6-8 week course of Optifast in potential donors to reduce 

body weight and fatty liver converting them to suitable live 

liver donors. Methods: Among 447 potential donors evaluated 

in our live donor liver transplant program between January 

2012 to December 2014, 47 potential donors with a BMI 

greater than 30, or evidence of significant steatosis on liver 

biopsy, were administered Optifast for 6-8 weeks during their 

evaluation for live liver donation. Outcomes including changes 

in BMI and degree of steatosis on liver biopsy before and 

after Optifast treatment were studied. Post-surgical outcomes 

for donors and recipients were also recorded, and compared 

to 91 live donors who did not receive Optifast and underwent 

donor hepatectomy within the study period. Results: Median 

BMI of Optifast potential donors (n=47) was 35.0 (IQR 33.1- 

37.3), compared to 25.9 (IQR 23.1-29.1) in all non-Optifast 

potential donors (n=400) [p


13 

A simple educational tool for reducing 30-day hospital 

readmissions in patients with decompensated cirrhosis 

Dennis Kumral, Michael W. Crothers, Stephen H. Caldwell, Zachary 

Henry; University of Virginia, Charlottesville, VA 

Background: Cirrhosis is a chronic disease with acute on 

chronic decompensating events that lead to hospitalization, 

similar to congestive heart failure (CHF) and chronic obstructive 

pulmonary disease (COPD). While patients with CHF and 

COPD have widespread targeted discharge pathways that 

help prevent readmissions, patients with cirrhosis have no 

standardized discharge programs. We developed a quality 

improvement initiative aimed at utilizing a standardized discharge 

pathway for patients with cirrhosis. Our goal was to 

reduce 30-day readmissions to our inpatient hepatology service 

by at least ten percent. Methods: In April 2015 we began a 

quality improvement discharge education program for patients 

with cirrhosis complicated by either fluid overload or hepatic 

encephalopathy. A dedicated hepatology nurse coordinator 

performed a teaching session with patients and their families 

prior to discharge and provided them with a patient-friendly 

cirrhosis management booklet created by the study team. All 

patients were provided with a digital scale and pill organizer 

if they did not already own one. Within 72-hours of discharge, 

a phone call was made to patients to review medications and 

reinforce teaching. Patients were monitored for readmission up 

to 30 days post discharge as the primary quality outcome. A 

control group of cirrhosis patients without a dedicated teaching 

program was chosen from hospital discharges during the same 

time period one year earlier. Thirty-day readmission rates were 

assessed for comparison. Results: A total of 20 patients with 

cirrhosis complicated by fluid overload and/or hepatic encephalopathy 

went through the discharge pathway and reached 

30 days post discharge at the time of this submission. We 

identified 25 control patients from the same time period one 

year earlier. There was no significant difference in age, gender, 

discharge MELD, discharge child pugh score, or length 

of stay between the two groups. The 30 day readmission rate 

in the study group was 25% compared to 62% in the control 

group, p=0.02. This correlates to a relative risk reduction of 

60% with a number needed to teach (NNT) of 2.7 to prevent 

one 30-day readmission. On multivariate logistic regression 

the only significant variable for predicting 30 day readmission 

was participating in the educational discharge pathway which 

was found to be protective, OR 0.22, p=0.02. Conclusions: 

Implementation of a dedicated educational discharge pathway 

with detailed teaching of cirrhosis management and a post-discharge 

follow-up phone call can reduce 30-day readmissions 

for patients with decompensated cirrhosis. 

Disclosures: 

Stephen H. Caldwell - Advisory Committees or Review Panels: Vital Therapy; 

Grant/Research Support: Genfit, Gilead Sciences, Immuron, Hyperion, Immuron, 

NGM 

The following authors have nothing to disclose: Dennis Kumral, Michael W. 

Crothers, Zachary Henry 

14 

Development of a Model to Predict Post-Surgical 

Unplanned Readmissions in Patients with Decompensated 

Cirrhosis 

Monica Schmidt 1 , Paul H. Hayashi 2 , Alfred S. Barritt 2 ; 1 UNC Liver 

Center & Gillings School of Global Public Health, University of 

North Carolina Chapel Hill, Chapel Hill, NC; 2 UNC Liver Center, 

Chapel Hill, NC 

Background:In the U.S., liver cirrhosis is expected to affect more 

than 1 million individuals by 2020 and many will require surgery. 

Prediction of 30-day readmission will be key in designing 

appropriate discharge planning and preventing readmissions. 

We developed a predictive model for 30-day readmission that 

is specific to patients with decompensated liver cirrhosis undergoing 

surgery. Methods:We used the National Surgical Quality 

Improvement Program data (NSQIP) from 2011-2013. Patients 

with cirrhosis were identified. Predictions were obtained for 

the model and the MELD score for comparison. The AUROC, 

cut-point, sensitivity/specificity and decision curve analysis 

are reported. Results: There were 5,879 patients with 739 

readmissions within 30-days (13%). Predictors of 30-day 

readmission were insulin dependent diabetes (OR 3.4 CI 1.6- 

7.3), discharged home (OR 3.4 CI 1.5-8.7), ASA class 4-life 

threatening (OR 4.7 CI 1.8-12.1), and days from surgery to 

discharge (OR 0.94 CI 0.91-0.97). The AUROC for the MELD 

score was 0.50 while our model reached 0.80 (p


15 

Effect of HCV treatment outcome on hospitalization rate: 

Chronic Hepatitis Cohort Study (CHeCS) 

Eyasu H. Teshale 1 , Jian Xing 1 , Anne C. Moorman 1 , Scott D. Holmberg 

1 , Stuart C. Gordon 2 , Loralee B. Rupp 2 , Mei Lu 2 , Philip R. 

Spradling 1 , Joseph A. Boscarino 3 , Connie M. Trinacty 4 , Mark A. 

Schmidt 5 , Fujie Xu 1 ; 1 CDC, Atlanta, GA; 2 Henry Ford Health System, 

Detroit, MI; 3 Geisinger Health System, Danville, PA; 4 Kaiser 

Permanente Hawaii, Honolulu, HI; 5 Kaiser Permanente Northwest, 

Portland, OR 

Background: Chronic hepatitis C virus (HCV) infection is associated 

with high all-cause hospitalization rates. Hospitalization 

rates increase with increasing severity of liver disease. Successful 

treatment of chronic HCV infection results in reduced 

morbidity and mortality. The objective of this study is to determine 

the association between HCV treatment outcome and 

hospitalization rate. Methods: We used data from HCV-infected 

patients seen in four large U.S. healthcare systems from 

2006-2012. We included persons who received HCV treatment 

during this time and who had follow up before and after 

completion of treatment. Treatment outcome was grouped as 

sustained virologic response (SVR) or treatment failure (TF). We 

determined hospitalization rates [per 100 person years (PY)] 

before and after treatment (SVR or TF) and excluded hospitalizations 

during treatment. Results: Between 2006 and 2012, 

1409 persons received HCV therapy; 680 (48.3%) achieved 

SVR and 729 (51.7%) had TF. The overall before treatment 

hospitalization rate was 13.2/100 PY; before treatment hospitalization 

rates for those who achieved SVR and those with 

TF were 10.6/100 PY and 16.0/100 PY, respectively. The 

after treatment hospitalization rate decreased to 5.6/100 PY 

for those who achieved SVR (p


17 

A Prospective Study of a Protocol to Reduce Early Readmission 

after Liver Transplantation 

Mark W. Russo 1 , Amit Mori 1 , David Levi 2 , Ruth Pierce 2 , Siddesh 

Besur 1 , Vincent P. Casingal 2 , Paul A. Schmeltzer 1 , Philippe J. 

Zamor 1 , Andrew Delemos 1 , Lon Eskind 2 ; 1 Hepatology, Carolinas 

Healthcare System, Charlotte, NC; 2 Transplant, Carolinas Medical 

Center, Charlotte, NC 

A Prospective Study of a Protocol to Reduce Early Readmission 

after Liver Transplant Introduction: Hospital readmission 

is an important marker of quality and delivery of care. Studies 

have evaluated risk factors for readmission after liver transplant 

(OLT) but few studies evaluated interventions to reduce 

readmission. Aim: To reduce 30 day readmission rates after 

OLT by implementing a prospective protocol with a multistep 

strategy. Methods: In October 2013 a comprehensive strategy 

was initiated to address early readmission after OLT. Core 

components of the protocol included revising criteria for readmission, 

alternatives to readmission, emphasized processes to 

improve patient teaching and discharge planning and expansion 

of outpatient services. 2014 served as the first full year 

the protocol was implemented. Readmission rates for 2014 

were compared to 2012 and 2013 with Fisher’s exact test and 

ANOVA. Logistic regression was used to control for potential 

confounding variables. Results: From January 1st 2012 thru 

December 31st 2014 167 adult OLTs were performed at our 

center with a mean biologic MELD of 21. The most common 

indications were HCV (35%), NAFLD (18%), HCC (17%), and 

ETOH (13%). The mean age of the study group was 54 y/o, 

63% were male. The mean ICU LOS was 3.4 days and hospital 

LOS 12.6 days. Over the study period 30 day readmission 

rate was 34% and decreased after implementing the readmission 

protocol from 40% in 2012 and 39% in 2013 to 19% 

in 2014, p=0.04. The most common reasons for readmission 

were biliary complications, infection, and rejection. Discharge 

year (before or after implementing protocol) remained significant 

after controlling for age, MELD score, indication, dialysis 

pre and post liver transplant, distance from transplant center, 

length of stay, PRBC transfusion, insurance and weekend discharge 

OR=0.40 [95% CI 017,0.94], p=0.04. The top factors 

identified as reducing readmission were expanding outpatient 

services and alternatives to inpatient readmission. Conclusions: 

Early readmission after liver transplantation can be reduced 

by expanding outpatient services and implementing alternative 

approaches to inpatient readmission. 

Disclosures: 

Mark W. Russo - Grant/Research Support: Merck, Salix; Speaking and Teaching: 

janssen, Gilead, ABBVIE, Salix 

Philippe J. Zamor - Grant/Research Support: AbbVie, Bristol Myers Squibb, 

Gilead, Merck & Co. ; Speaking and Teaching: AbbVie, Bristol Myers Squibb, 

Janssen 

The following authors have nothing to disclose: Amit Mori, David Levi, Ruth 

Pierce, Siddesh Besur, Vincent P. Casingal, Paul A. Schmeltzer, Andrew Delemos, 

Lon Eskind 

18 

Reduced Work Productivity (WP), Absenteeism and 

Presenteeism of Patients Infected with Hepatitis C Virus 

(HCV) are Independently Predicted by Physical Component 

of Patient-Reported Outcomes (PROs) 

Zobair M. Younossi 1,2 , Maria Stepanova 3 , Linda Henry 3 , Issah 

Younossi 3 , Ali A. Weinstein 3 , Fatema Nader 1 , Sharon L. Hunt 3 ; 

1 Center For Liver Disease, Department of Medicine, Inova Fairfax 

Hospital, Falls Church, VA; 2 Betty and Guy Beatty Center for Integrated 

Research, Inova Health System, Falls Church, VA; 3 Center 

for Outcomes Research in Liver Disease, Washington, DC 

Background: Hepatitis C virus (HCV) infection is associated with 

significant impairment of health-related quality of life (HRQOL) 

and other PROs. Additionally, HCV infected patients are known 

to have reduced work productivity (WP), both in terms of presenteeism 

(impairment in work productivity while working) and 

absenteeism (productivity loss due to absence from work). Aim: 

The aim of this study was to identify PROs which are predictive 

of WP in untreated HCV-infected patients. Methods: We 

analyzed data from HCV patients prior to intitaion of anti- 

HCV regimens in a clinical trial setting. All patients had completed 

4 PRO questionnaires (CLDQ-HCV, SF-36, FACIT-F and 

WPAI:SHP). In subjects who reported being employed, WP 

and its absenteeism and presenteeism components were calculated 

using WPAI:SHP instrument. Independent PRO predictors 

of WP, absenteeism and presenteeism were assessed using 

multiple linear regression. Results: Of 4,121 HCV patients who 

completed pre-treatment WPAI:SHP, 2,480 (60.2%) reported 

to be employed, and of those, 2,121 had completed all PRO 

questionnaires before treatment initiation. The study cohort was 

51.1±9.6 years old, 16.7% cirrhotic, 76.1% HCV genotype 1, 

and 62.3% treatment-naïve. Average baseline impairment in 

WP was 11.0%, including 8.3% (75% of WP impairment) of 

presenteeism and 2.7% (25% of WP impairment) of absenteeism. 

Using multivariate regression analysis, WP was predicted 

by the activity/energy domain of CLDQ-HCV (beta=4.55±0.56 

per 1 point, p


19 

Administration of an Ileal Apical Sodium-dependent Bile 

Acid Transporter Inhibitor Protects Against Non-alcoholic 

Fatty Liver Disease in High Fat Diet-fed Mice 

Anuradha Rao 1 , Astrid Kosters 1 , Jamie Mells 1 , Bradley T. Keller 4 , 

Hong Yin 2 , Sophia Banton 2 , Shuzhao Li 2 , Dean P. Jones 2 , Hao 

Wu 3 , Paul Dawson 1 , Saul J. Karpen 1 ; 1 Pediatrics, Emory University, 

Atlanta, GA; 2 Medicine, Emory University, Atlanta, GA; 3 Public 

Health, Emory University, Atlanta, GA; 4 Vasculox, Inc, St. Louis, 

MO 

Non-alcoholic fatty liver disease (NAFLD) is the most common 

chronic liver disease in the Western world. The mechanisms 

underlying NAFLD development and progression are not 

fully elucidated, and safe and effective NAFLD therapies are 

needed. Bile acids (BA) and their receptors have important 

roles in regulating whole body metabolism, including multiple 

hepatic targets coordinating lipid handling. We hypothesized 

that interruption of the enterohepatic BA circulation via 

a non-absorbable Apical Sodium-dependent BA Transporter 

(ASBT) inhibitor (ASBTi; SC435) would modify signaling in 

the gut-liver axis and diminish the development of NAFLD in 

High Fat Diet (HFD) fed mice. Methods: Male C57Bl/6 mice 

(n=12-16/group) were fed for 16 weeks with A) chow, B) HFD 

composed of 45% fat and 0.2% cholesterol, with 4% sucrose 

water (HFD), and C) HFD plus ASBTi (SC435; 60 ppm; HFD/ 

ASBTi). Body weight, food intake, glucose and insulin tolerance, 

liver histology, and liver lipids were measured. Molecular 

mechanisms were examined using RNASeq, metabolomics, 

and real-time PCR analyses. Results: In HFD versus HFD/ASBTi 

mice, administration of the ASBTi increased fecal BA excretion, 

and mRNA expression for colonic Ibabp (7.0±2.0 fold; 

p


Inflammasomes are new molecular platforms that respond to 

microbial products through the synthesis of pro-inflammatory 

cytokines. The Nlrp3 inflammasome activation is known to 

determine Il-1β and Il-18 synthesis and release, thereby modulating 

cell biology and eliciting pro-inflammatory signals in 

different cell types. We sought to verify whether the inflammasome 

is activated in sclerosing cholangitis and which are its 

effects in reactive cholangiocytes. Methods: Expression levels 

of the NLRP3 inflammasome were tested in cholangiocytes of 

normal and cholestatic livers. The effects of Nlrp3 activation 

induced by incubation with LPS/ATP were studied in vitro in 

normal and siRNA Nlrp3 knockdown cholangiocytes. In vivo, 

wild type (WT) and Nlrp3 A350VneoR 

(Nlrp3 -/- ) mice were fed 

with 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC, a model 

of sclerosing cholangitis) for 4 weeks. Results: Expression of 

Nlrp3 and its components is increased in in cholangiocytes 

of mice subjected to DDC and in patients affected by PSC 

compared to normal conditions. LPS/ATP-induced activation 

of Nlrp3 in cholangiocytes stimulates the expression of Il-18 

and Il-6, but not of Il-1β. Nlrp3 activation also significantly 

decreased the expression of Zonulin-1 and E-cadherin but 

had no effect on cholangiocyte proliferation. The effects of 

LPS/ATP-induced activation of Nlrp3 in cholangiocytes were 

neutralized by knockdown of Nlrp3 by siRNA. In vivo, the 

increases in the liver CK-19-positive parenchyma induced by 4 

week DDC in WT animals were reduced in Nlrp3 -/- mice and 

expression of Zonulin-1 tended to be re-established. Conclusions: 

Nlrp3 is expressed in reactive cholangiocytes, both in 

murine models and in PSC. The activation of Nlrp3 leads to 

the synthesis of pro-inflammatory cytokines and influences the 

epithelial integrity of cholangiocytes. These findings suggest 

that microbial products may participate to the development of 

certain cholangiopathies by activating the inflammasome in 

cholangiocytes and altering the barrier function of the biliary 

epithelium. 

Disclosures: 

The following authors have nothing to disclose: Luca Maroni, Laura Agostinelli, 

Stefania Saccomanno, Eleonora Mingarelli, Chiara Rychlicki, Samuele De Minicis, 

Jesus M. Banales, Antonio Benedetti, Gianluca Svegliati Baroni, Marco 

Marzioni 

22 

Taurocholate Activates YAP via Sphingosine-1 Phosphate 

Receptor 2 in Cholangiocytes 

Kesong Peng 1 , Yunzhou Li 2 , Runping Liu 1,2 , Jing Yang 1 , Yongqing 

Wang 3,1 , Guang Liang 4 , Phillip B. Hylemon 1,2 , Huiping Zhou 1,2 ; 

1 Microbiology and Immunology, Virginia Commonwealth University, 

Richmond, VA; 2 McGuire Veterans Affairs Medical Center, 

Richmond, VA; 3 Nanjing Medical University, Nanjing, China; 

4 School of Pharmacy, Wenzhou Medical University, Wenzhou, 

China 

Introduction: The transcription co-activators YAP/TAZ are the 

core components of the Hippo pathway. Recent studies demonstrated 

that activation of several G protein coupled receptors 

(GPCRs), including sphingosine-1-phosphate receptor 2 

(S1PR2), causes dephosphorylation and nuclear translocation 

of YAP. Cholangiocytes are continuously exposed to high concentrations 

of conjugated bile acids. Obstruction of bile flow 

is a potent stimulus for cholangiocyte proliferation. We have 

recently reported that taurocholate (TCA) promotes cholangiocyte 

proliferation via activation of S1PR2, which is the predominant 

S1P receptor in cholangiocytes. Recently, it was reported 

that bile acids promote liver carcinogenesis through activation 

of YAP. However, whether activation of S1PR2 by conjugated 

bile acids induces YAP activation in cholangiocytes has not 

been examined and is the focus of this study Methods: A mouse 

cholangiocyte cell line was used for in vitro studies. Bile duct 

ligation (BDL) mouse models were used to examine the role 

of S1PR2 in YAP activation in vivo. A chemical antagonist of 

S1PR2, JTE-013, was used to inhibit S1PR2 activation. S1P 

was used as positive control. YAP activation was determined 

by Western blot analysis. The mRNA levels of the YAP target 

genes were determined by real-time RT-PCR. Results: Both 

TCA- and S1P-induced cell migration and proliferation were 

inhibited by JTE-013 in cholangiocytes. TCA- and S1P-induced 

activation of YAP was indicated by decreasing of phosphorylated 

YAP in cytosol and increasing of nuclear translocation of 

YAP, which was markedly inhibited by JTE-013. In addition, 

TCA and S1P also significantly up-regulated the expression 

of the down-stream target genes of YAP including: IQGAP1, 

cyclin D1, p21 and CTGF (connective tissue growth factor), 

in cholangiocytes. YAP activation was significantly reduced in 

BDL S1PR2 -/- mice as compared to wild-type mice. Discussion/ 

Conclusion: Elevated conjugated bile acid levels are correlated 

with cholangiocyte proliferation and cholangiocarcinoma by 

unknown mechanism(s). YAP-activation has been linked to 

tumour growth. This study identified TCA/S1PR2/YAP/TAZ as 

a cell signalling pathway involved in TCA-mediated cholangiocyte 

proliferation and possibly transformation. 

Disclosures: 

The following authors have nothing to disclose: Kesong Peng, Yunzhou Li, Runping 

Liu, Jing Yang, Yongqing Wang, Guang Liang, Phillip B. Hylemon, Huiping 

Zhou 

23 

SRT1720 protects against cholestatic liver injury in mice 

by altering hepatic bile acid composition and enhancing 

urinary excretion of bile acids 

Supriya Kulkarni 1 , Carol J. Soroka 1 , Lee R. Hagey 2 , James L. 

Boyer 1 ; 1 Liver Center, Internal Medicine, Digestive Diseases, Yale 

University School of medicine, New Haven, CT; 2 Division of Gastroenterology, 

Department of Medicine, University of California at 

San Diego, La Jolla, CA 

BACKGROUND: Sirtuin1 (SirT1, mammalian homolog of S. Cerevisiae 

enzyme Sir2) is a critical transcriptional and transactivational 

regulator of murine Fxr. Liver specific knockout of Sirt1 

deregulates Fxr activity. Our previous studies demonstrated that 

bile duct ligation (BDL), or cholic acid (CA) feeding decrease 

hepatic Sirt1 expression and that a Sirt1 activator SRT1720 

improves cholestatic liver injury. AIM: To determine the mechanisms 

by which Sirt1 activation may alleviate cholestatic liver 

injury. METHODS: C57Bl/6 mice (n=11-13, male, 8-9 weeks 

old) were fed standard rodent chow or chow supplemented 

with 1% CA for 5 days. Both groups (n=5-6) were administered 

either vehicle or Sirt1 activator, SRT1720 (50mg/kg/ 

day) for duration of the diet. Liver injury (ALT, ALP), BA levels in 

plasma, liver, ileum, urine and the bile acid pool size as well 

as gene and protein expression of BA transporter and synthesis 

genes were determined. Immunoprecipitation assays were 

used to determine Sirt1 acetylation and Fxr activity. RESULTS: 

Sirt1 mRNA, protein and acetylation activity were significantly 

reduced (75%) in CA fed mouse livers. SRT1720 administration 

in CA fed mice reduced plasma ALT (40%) and plasma 

BA (50%) levels while hepatic and total BA pool remained 

unchanged. SRT1720 significantly increased tetrahydroxylated 

BA by 1.96x and decreased di-hydroxylated BA fraction 

to 21% over CA fed mice and increased Cy2b10 (1.75x) 

expression. SRT1720 increased ileal Fgf15 (30x) and hepatic 

Fgfr4 (2X), decreased hepatic Cyp7a1 (99%), 27a1 (75%) 

expression. SRT1720 also increased renal Mrp2 and Mrp4 

expression (2-4.5x) along with increased urinary BA concen-


tration (2x) relative to CA fed mice, thus increasing BA excretion 

in the urine. Hepatic Jnk was activated by CA feeding, 

a finding that that was reversed by SRT1720 administration 

which restored Sirt1 and Fxr mRNA, protein and activity levels 

to normal. CONCLUSION: Sirt1 protein levels and acetylation 

activity are decreased in CA feeding model of cholestasis Sirt1 

activator SRT1720 decreased cholestatic liver injury by several 

mechanisms: SRT1720 increased the hepatic concentration of 

hydrophilic hydroxylated BA likely via increased expression 

of Cyp2b10. In addition, SRT1720 decreased BA synthesis 

via the Fgf15-Fgfr-Cyp7a1 pathway and finally SRT1720 

decreased serum bile acid levels by increasing renal Mrp2 

and Mrp4 expression and urinary BA excretion. These observations 

could be attributed to SRT1720 mediated reversal of Jnk 

activation and hence restoration of Sirt1 and Fxr expression in 

the CA fed mice. 

Disclosures: 

James L. Boyer - Advisory Committees or Review Panels: Pfizer; Consulting: 

abbvie 

The following authors have nothing to disclose: Supriya Kulkarni, Carol J. 

Soroka, Lee R. Hagey 

24 

FXR-β-catenin complex and bile acid homeostasis: therapeutic 

implications in cholestatic liver disease 

Kari Nejak-Bowen, Satdarshan (Paul) S. Monga; University of Pittsburgh, 

Pittsburgh, PA 

Cholestatic liver diseases are characterized by bile acid (BA) 

accumulation in the liver, which can lead to inflammation 

and fibrosis, and eventually result in end-stage liver disease. 

We utilize the bile duct ligation (BDL) model and identify a 

dramatic reduction in liver injury and fibrosis in liver-specific 

β-catenin knockout (KO) mice. This is due to a previously unrecognized 

role of β-catenin in regulation of BA synthesis and 

transport through association with farnesoid X receptor (FXR). 

We confirm this association through immunoprecipitation (IP) 

of FXR and β-catenin from wild-type (WT), β-catenin KO, and 

FXR KO livers and showed association of the two proteins in 

WT but not in either KO. Further, the association with FXR 

occurs at the C-terminal of β-catenin, since IP of β-catenin and 

FXR from HepG2 cells reveals association with both the fulllength 

and N-terminal truncated β-catenin forms. Additionally, 

the β-catenin-FXR pool is unresponsive to BA stimulation as 

demonstrated by IP studies on GW4064-treated Hep3B cells. 

To address the functional relevance of this complex, we show 

that absence of β-catenin allows FXR activation to occur more 

readily, both in vitro and in vivo. In fact, a more pronounced 

FXR activation in KO is the major reason of decreased total 

hepatic BA and injury after BDL. Conversely, when Hep3B cells 

were transfected with S33Y-mutated stable β-catenin, we find 

increased FXR-β-catenin association by IP. This coincides with 

loss of FXR-RXRα association, which normally causes activation 

of SHP and suppresses BA synthesis. Expression of S33Y-βcatenin 

also abrogates the GW4064-induced increase in SHP 

reporter activity seen in Hep3B cells treated with control plasmid, 

confirming the inhibitory role of this association. This was 

validated in HepG2 cells, which show less SHP induction after 

β-catenin silencing than Hep3B cells, as the truncated/stable 

form of β-catenin is still present in these cells after β-catenin suppression. 

The complex interplay between β-catenin, FXR and 

RXRα was further validated when overexpression of RXRα led 

to a switch in association of FXR from β-catenin to RXRα. Thus, 

we have identified a novel FXR-β-catenin complex and propose 

its manipulation may provide a novel therapeutic opportunity 

for alleviating BA-associated hepatic injury during cholestasis. 

Disclosures: 

Satdarshan (Paul) S. Monga - Advisory Committees or Review Panels: Abbvie; 

Grant/Research Support: Dicerna Pharmaceuticals 

The following authors have nothing to disclose: Kari Nejak-Bowen 

25 

HIV and HCV co-infection in hepatocytes and stellate 

cells reveals cooperative transcriptional activation 

between viruses and cell types 

Shadi Salloum 1 , Cynthia Brisac 1 , Rohit Jindal 2 , Shyam Sundhar 

Bale 2 , Nadia Alatrakchi 1 , Annie Kruger 1 , Anna Lidofsky 1 , Martin 

L. Yarmush 2 , Raymond T. Chung 1 ; 1 GI, MGH, Boston, MA; 2 Center 

for Engineering in Medicine, Shriner, Boston, MA 

Background: HIV/HCV co-infection accelerates progression to 

liver fibrosis, a major public health problem. Currently, the 

molecular and cellular mechanisms underlying progression of 

this dynamic disease remain incompletely understood. We previously 

demonstrated, in monoculture experiments, that HCV 

and HIV independently led to the production of profibrogenic 

TGFβ through an increase in reactive oxygen species (ROS) 

and NFkB activation in hepatocytes as well as in hepatic stellate 

cells (HSCs), all of which are associated with the development 

of hepatic fibrosis. Aim: We sought to create a co-culture 

model combining hepatocytes and HSCs to effectively simulate 

HCV and HIV co-infection in vitro. This model will be of 

great utility to evaluate the mechanistic contributions of HIV 

and HCV infection to hepatic fibrogenesis in a multicellular 

context. Methods: We first constructed stable monoclonal 

green fluorescent protein (GFP) reporter cells expressing functional 

antioxidant response element (ARE), NFkB, and SMAD3 

driven reporter transgenes, which enables these transcriptional 

regulators to be studied dynamically in living cells. Then, we 

adapted the reporter cells to create a model of co-infection 

that allows investigation of transcriptional dynamics associated 

with HIV/HCV co-infection. Finally, we used the reporter cells 

to create co-cultures of hepatocytes and stellate cells to further 

investigate the effect of HIV/HCV co-infection. Results: The 

efficiency of reporter cell lines was validated by using positive 

controls which activated 85% to 60% of total cells versus only 

2 to 10% mock activated cells. In In monoculture, HIV and 

HCV induced ARE, and SMAD3 in both cell lines (suggesting 

an increase in cellular oxidative stress and TGF-β production). 

In contrast, NFkB was activated only in Huh7.5.1 but not in 

LX2 cells. Co-infection with HCV and HIV led to a significant 

increase (2-3 fold) in the activation of these reporters in comparison 

to monoinfection. The co-culture model confirmed HCV 

and/or HIV effects, on SMAD, ARE, and NFkB activation. 

Interestingly, the viruses’ effects on these reporters increased 

additively compared to monoculture in Huh7.5.1 cells. Conclusions: 

HIV accentuates the HCV-related profibrogenic program 

in HSCs and hepatocytes through initial ROS induction, NF-kB, 

and TGF-B1 upregulation. Co-culture studies indicated additive 

effects compared with monoculture. Our novel co-culture 

reporter cell model represents an efficient system for studying 

transcriptional responses and has the potential to provide 

important insights into the progression of fibrosis liver disease, 

as well as study of potential therapeutic interventions. 

Disclosures: 

Raymond T. Chung - Grant/Research Support: Gilead, Mass Biologics, Abbvie, 

Merck, BMS 

The following authors have nothing to disclose: Shadi Salloum, Cynthia Brisac, 

Rohit Jindal, Shyam Sundhar Bale, Nadia Alatrakchi, Annie Kruger, Anna 

Lidofsky, Martin L. Yarmush


26 

Elevation of Galectin-9 is associated with the cytotoxicity 

of NK cells in chronic hepatitis 

Akira Nishio, Tomohide Tatsumi, Takatoshi Nawa, Takahiro 

Suda, Atsuo Takigawa, Tadashi Kegasawa, Yoshiki Onishi, Seiichi 

Tawara, Teppei Yoshioka, Satoshi Aono, Minoru Shigekawa, 

Hayato Hikita, Ryotaro Sakamori, Naoki Hiramatsu, Tetsuo Takehara; 

Department of Gastroenterology and Hepatology, Osaka 

University Graduate School of Medicine, Suita, Japan 

Background & Aims: NK cells play an essential role in the 

pathogenesis of chronic hepatitis C (CHC). NK cells are activated 

in CHC and activated NK cells exhibit elevated cytotoxicity 

leading to liver injury. Recently, the activated NK cells 

contribute to exhaustion of virus-specific T cells, which is associated 

with persistent viral infection. However, the mechanism 

of NK cell activation has not been clearly understood. The 

immunoregulatory protein Galectin-9 (Gal-9) is elevated in the 

serum and liver in CHC patients, suggesting that Gal-9 might 

be involved in the pathogenesis of CHC. In this study, we investigated 

the effect of Gal-9 on the activation of NK cells in CHC. 

Methods: Serum Gal-9 levels were analyzed in 39 healthy 

donors, 50 CHC patients and 24 IFN-treated patients achieving 

sustained viral response (SVR) by ELISA. The cytotoxicity of 

NK cells and Gal-9 levels in the supernatant were evaluated by 

flow cytometry and ELISA. Immunofluorescence staining was 

performed in CHC liver tissue. Results: Serum Gal-9 levels in 

CHC patients were significantly higher than those in healthy 

donors, and those in SVR patients were significantly lower 

than those in CHC patients (median, 0-90 percentile, CHC: 

146 pg/ml, 0-508, healthy donors: 0, 0-55.4, SVR: 53.6, 

0-185.6). In CHC patients, serum levels of Gal-9 in elevated 

ALT patients were significantly higher than those in normal ALT 

patients, suggesting that elevated Gal-9 might be associated 

with liver injury. Serum Gal-9 levels were not associated with 

HCV viral loads. In vitro study revealed that addition of recombinant 

Gal-9 resulted in significant increase of the cytotoxicity 

of naïve NK cells in a dose dependent manner, independent of 

Tim-3. Gal-9-activated NK cells showed the cytotoxicity against 

CD4+ or CD8+ T cells, which might contribute to persistent 

infection by depleting T cell. We found that a large amount of 

Gal-9 production was induced in the co-culture of PBMCs with 

JFH-1/Huh7.5.1 cells, but not with uninfected Huh7.5.1 cells. 

A transwell system revealed that cell-cell contact was required 

for Gal-9 production. In vitro depletion study revealed CD14+ 

monocytes were required for Gal-9 in the co-culture, and both 

Gal-9 and CD14 positive cells were observed in immunofluorescence 

staining, indicating that Gal-9 was produced by 

Kupffer cells in the liver. The cytotoxicity of NK cells was significantly 

elevated when co-cultured with JFH-1 cells compared 

with control. Conclusion: Monocytes-derived Gal-9 induces the 

cytotoxicity of NK cells, which might be involved in liver injury 

and persistent HCV infection. These results suggest the role of 

Gal-9 in the pathogenesis of chronic hepatitis C. 

Disclosures: 

Hayato Hikita - Grant/Research Support: Bristol-Myers Squibb 

Tetsuo Takehara - Grant/Research Support: Chugai Pharmaceutical Co., MSD 

K.K., Bristol-Meyer Squibb, Mitsubishi Tanabe Pharma Corparation, Toray Industories 

Inc. ; Speaking and Teaching: MSD K.K., Bristol-Meyer Squibb, Janssen 

Pharmaceutical Companies 

The following authors have nothing to disclose: Akira Nishio, Tomohide Tatsumi, 

Takatoshi Nawa, Takahiro Suda, Atsuo Takigawa, Tadashi Kegasawa, Yoshiki 

Onishi, Seiichi Tawara, Teppei Yoshioka, Satoshi Aono, Minoru Shigekawa, 

Ryotaro Sakamori, Naoki Hiramatsu 

27 

De novo formation and maturation of hepatitis C virus 

replication membranes visualized by pulse-chase imaging 

Hongliang Wang 1 , Andrew W. Tai 1,2 ; 1 Internal Medicine, University 

of Michigan, Ann Arbor, MI; 2 VA Ann Arbor Healthcare 

System, Ann Arbor, MI 

Hepatitis C virus (HCV) replicates on an altered host membrane 

structure called the “membranous web” (MW). Little is known 

regarding temporal events in MW formation and association 

with putative sites of virus assembly at or near lipid droplets 

(LDs). For example, it is not known whether membranous webs 

are stable structures that are continually renewed by newly 

synthesized viral proteins, or whether newly synthesized viral 

proteins are directed to form new MWs de novo. The HCV 

nonstructural protein NS5A is believed to be a component of 

MWs. We have developed pulse-chase fluorescent labeling 

and imaging approaches that allow us to discriminate ‘old’ 

from ‘new’ NS5A-positive membranous structures (NS5A foci). 

With this system, we find that ‘new’ NS5A-positive foci form at 

sites distinct from ‘old’ NS5A foci, supporting a model of de 

novo MW formation instead of renewal of previously formed 

MWs. We also find that the phosphatidylinositol 4-kinase 

PI4KA and oxysterol-binding protein, which are known to be 

required for normal MW morphogenesis, are required to maintain 

the normal morphology of both ‘old’ and ‘new’ foci, and 

that these two proteins appear to play a role in de novo NS5A 

focus formation. Finally, we observe increasing colocalization 

of NS5A-positive foci with HCV core protein and LDs over time. 

Overall, these data support a model in which HCV MWs are 

formed de novo rather than renewed. We hypothesize that 

newly-formed NS5A foci undergo a process of maturation and 

movement towards sites of putative virion assembly at or near 

LDs. This may have implications for our understanding of how 

replication membranes are formed for other positive-sense RNA 

viruses. 

Disclosures: 

The following authors have nothing to disclose: Hongliang Wang, Andrew W. 

Tai 

28 

Rapid normalization of antiviral and pro-inflammatory 

transcriptional signatures in peripheral blood of patients 

and livers of humanized mice treated with DAAs 

Matthew A. Burchill 1 , Lucy Golden-Mason 1 , Megan Wind-Rotolo 2 , 

Michael Gale 3 , Hugo R. Rosen 1 ; 1 GI-Hepatology, University of 

Colorado-Denver, Aurora, CO; 2 Exploratory Clinical and Translational 

Research, Bristol-Myers Squibb, Lawrenceville, NJ; 3 Immunology, 

University of Washington, Seattle, WA 

Chronic HCV infection results in sustained immune activation 

in both the periphery and hepatic tissue. Recently, novel 

direct acting antiviral (DAA) regimens have been shown to 

induce rapid and sustained clearance of HCV. DAAs have 

provided the unique opportunity to determine whether successful 

treatment-induced eradication of viral RNA normalizes 

the dysregulated antiviral innate immune response in patients 

chronically infected with HCV. Methods: We used both a 

prospective cohort of patients and a novel humanized mouse 

model. Results: First, in 35 patients receiving DAAs, we found 

that a regimen of daclatasvir, asunaprevir, and BMS-791325 

induced not only rapid viral clearance, but also a re-setting 

of antiviral responses in the peripheral blood. Specifically, 

we observed that following treatment with DAAs, there was 

a significant reduction in IFITM1 and the chemokines CXCL10


and CXCL11 in the peripheral blood by Affymetrix microarray 

and confirmatory RT-PCR. Next, using a novel humanized 

mouse model (FRG), we assessed the transcriptional changes 

that occur in the hepatic tissue following DAA treatment. After 

establishing a stable chronic HCV infection in the mice, they 

were treated with daclatasvir, asunaprevir, and sofosbuvir that 

induced the clearance of circulating HCV within 48 hours. 

Despite the rapid clearance of circulating viral particles, we 

observed HCV NS3 protein within infected hepatocytes 14 

days after initiation of DAA therapy. The presence of residual 

NS3 protein did not affect the blunting of the expression 

of proinflammatory responses as we observed a significant 

reduction in the chemokines CXCL10 and CXCL11 in the liver 

of treated mice. These data suggested that the transcriptional 

changes that occur in the peripheral blood of patients treated 

with DAAs may partially reflect intrahepatic transcriptional 

changes. However, in livers of DAA-treated humanized mice 

we observed a restoration of mitochondrial anti-viral signaling 

protein (MAVS) and an increase in the protein IFITM1, the 

latter a tight junction protein typically induced only by IFN. 

Conclusions: These findings highlight how viral clearance elicited 

by DAA treatment leads to the rapid restoration of innate 

signaling moieties that are suppressed by HCV infection. Collectively, 

our data demonstrate that the rapid viral clearance 

following treatment with DAAs results in the rebalancing of 

innate antiviral response in both the peripheral blood and the 

liver as well as enhanced antiviral signaling within previously 

infected hepatocytes. 

Disclosures: 

Megan Wind-Rotolo - Employment: Bristol-Myers Squibb; Stock Shareholder: 

Bristol-Myers Squibb 

The following authors have nothing to disclose: Matthew A. Burchill, Lucy Golden-Mason, 

Michael Gale, Hugo R. Rosen 

29 

Activation of Hepatic Stellate Cells Drives the Rapid 

Development of Liver Fibrosis During Acute HCV Infection 

in HIV-infected Men 

Robert D. Gillies 1,2 , Daniel S. Fierer 1 , Marcus Yip 1,2 , Scott L. Friedman 

3 , Andrea D. Branch 3 , M. Isabel Fiel 4 ; 1 Infectious Diseases, 

Mount Sinai School of Medicine, New York, NY; 2 Monash University, 

Melbourne, VIC, Australia; 3 Liver Diseases, Icahn School 

of Medicine at Mount Sinai, New York, NY; 4 Pathology, Icahn 

School of Medicine at Mount Sinai, New York, NY 

Background The international epidemic of sexually-transmitted 

HCV infection among HIV-infected men is characterized by 

rapid development of liver fibrosis of unknown mechanism(s). 

The purpose of this study is to determine whether increased 

fibrogenesis during acute HCV infection is associated with 

activation of hepatic stellate cells (HSC), the principal collagen-producing 

cell in liver injury. Methods Liver biopsies were 

evaluated from HIV-infected men with acute and recent HCV 

infection in New York City. Date of HCV diagnosis was set 

as the date of the first-noted ALT elevation. The presence of 

alpha-smooth muscle actin (ASMA), indicative of activated 

HSC, was determined by counting stained foci in 10 random 

lobular areas per high-powered field (hpf) (40x magnification) 

on slides immunostained with antibodies to ASMA. Fibrosis 

stage (Scheuer, 0-4) was determined by examining Masson 

trichrome-stained slides. Collagen-proportionate area (CPA) 

was determined by computer digital image analysis of Sirius 

red-stained slides. The effect of time to biopsy on these three 

measurements was examined as a dichotomous variable, using 

1 year after HCV diagnosis. The Chi-Square test 

for trend was used to compare Scheuer stage of fibrosis; and 

the Mann-Whitney U Test was used to compare the median 

CPA and median activated HSC/hpf between groups. Results 

Liver biopsies from 20 men were evaluated. Median age at 

HCV diagnosis was 42 years, with a median of 9 years since 

HIV diagnosis; and median CD4 count was 501 cells/uL (all 

had >200). The median number of activated HSC/hpf was 

significantly higher in biopsies obtained 1 year after HCV diagnosis (p = 0.002, Table), indicating 

greater HSC activation early in HCV infection and lower 

HSC activation later in infection. Despite this lower activation 

of HSC in biopsies >1 year after HCV diagnosis, fibrosis 

stage (p = 0.01), and CPA (p = 0.06) were both higher in 

those with biopsy >1 year after HCV infection. Conclusion The 

rapid development of fibrosis that occurs in HIV-infected men 

during the first year after acute HCV infection is associated 

with ASMA-staining fibrogenic myofibroblasts. The number of 

activated HSC was lower after the first year of HCV infection 

while the stage of fibrosis was higher, indicating that once 

fibrogenesis has been set in motion, this process continues, 

although possibly at a slower pace. This rapid development of 

fibrosis can therefore not be considered to be benign. 

Disclosures: 

Daniel S. Fierer - Stock Shareholder: Gilead 

Scott L. Friedman - Advisory Committees or Review Panels: Pfizer Pharmaceutical; 

Consulting: Conatus Pharm, Exalenz, Genfit, Exalenz Biosciences, Eli Lilly PHarmaceuticals, 

Fibrogen, Boehringer Ingelheim, Nitto Corp., Immune Therapeutics, 

Synageva, Roche/Genentech Pharmaceuticals, DeuteRx, Abbvie, Novartis, 

RuiYi, Kinemed, Sanofi Aventis, Takeda Pharmaceuticals, Nimbus Therapeutics, 

Bristol Myers Squibb, Astra Zeneca, Sandhill Medical Devices, Galmed, Northern 

Biologics, Enanta Pharmaceuticals, Regado Bioscience, Raptor Pharmaceuticals, 

Teva Pharmaceuticals, Zafgen Pharmaceuticals, Merck Pharmaceuticals, 

Debio Pharmaceuticals; Grant/Research Support: Galectin Therapeutics, Tobira 

Pharm; Stock Shareholder: Angion Biomedica, Intercept Pharma 

Andrea D. Branch - Grant/Research Support: Gilead, Janssen 

The following authors have nothing to disclose: Robert D. Gillies, Marcus Yip, 

M. Isabel Fiel 

30 

Alcohol increases the production of hepatitis C virus 

(HCV) lipo-viro-particles in primary human hepatocytes 

Veronique Pene 1 , Céline Hernandez 1 , Etienne Blanc 2 , Lynda 

Aoudjehane 3,4 , Béatrice Le Grand 2 , Arnaud Carpentier 1 , Jean- 

François Méritet 1,5 , Filomena Conti 4,6 , Yvon Calmus 4,6 , Hélène 

Rouach 2 , Philippe Podevin 1,7 , Arielle R. Rosenberg 1,5 ; 1 EA 4474 

“Hepatitis C Virology”, Université Paris Descartes, Paris, France; 

2 UMRS 1124 “Toxicology Pharmacology and Cellular Signaling”, 

Université Paris Descartes, Inserm, Paris, France; 3 Human HepCell, 

Hôpital Saint-Antoine, Paris, France; 4 UMRS 938 “CDR Saint-Antoine”, 

Inserm, Paris, France; 5 Service de Virologie, AP-HP, Hôpital 

Cochin, Paris, France; 6 Unité de Transplantation Hépatique, AP-HP, 

Hôpital Pitié-Salpêtrière, Paris, France; 7 Centre de Référence en 

addictologie, AP-HP, Hôpital Pitié-Salpêtrière, Paris, France 

BACKGROUND & AIM: Alcoholism is a major aggravating 

factor of HCV infection, yet the molecular mechanisms of this 

comorbidity remain elusive. Clinical studies found higher viral 

loads in patients with chronic excessive alcohol consumption. 

However, the so-called “viral load” reflects a wide heterogeneity 

of HCV particles, the most infectious ones being those 

of lowest buoyant density due to their association with verylow-density 

lipoproteins (VLDL, the apolipoprotein B-containing 

triglyceride-rich lipoproteins secreted by hepatocytes) in hybrid 

structures termed lipo-viro-particles (LVP). With the aim of investigating 

the impact of alcohol on HCV infectious cycle, we took


advantage of our HCV culture system in primary human adult 

hepatocytes (PHH), which, contrary to the widely used hepatocarcinoma-derived 

Huh-7 sublines, retain the liver metabolism 

of ethanol and secrete authentic VLDL and LVP. METHODS: 

PHH were infected with the HCV strain JFH1 or a chimeric 

virus derived thereof and treated with increasing doses of ethanol 

(0-100 mM) for 2 weeks. Cultures were monitored for 

HCV genome replication (negative strand RT-qPCR), viral load 

(clinically used test), production of infectious virus (titration by 

focus-formation assay), and VLDL secretion (apolipoprotein B 

ELISA). The density of viral particles was assessed by isopycnic 

iodixanol ultracentrifugation. RESULTS: Ethanol exposure of 

HCV-infected PHH caused a time- and dose-dependent increase 

in the viral load, comparable to that reported in clinical studies. 

Most strikingly, it caused an even greater increase in the 

infectious titer but did not significantly affect the viral genome 

replication, thus pointing to an effect on virus morphogenesis. 

This effect was not seen in Huh-7.5.1 cells treated in parallel, 

suggesting that it involves the liver metabolism of ethanol. The 

higher specific infectivity of HCV particles produced by PHH 

in the presence of ethanol correlated with lower mean buoyant 

density, consistent with triglyceride enrichment. Finally, in 

either HCV-infected or naïve PHH, addition of ethanol caused 

a time-dependent increase in VLDL secretion. CONCLUSION: 

This study in the relevant PHH model reveals that ethanol via its 

metabolites increases the production of HCV particles of lowest 

buoyant density and highest infectivity, i.e., LVP, most likely 

due to the impact of ethanol on triglyceride metabolism that 

results in increased VLDL secretion. Drugs targeting this host 

metabolic pathway may be useful in difficult-to-treat alcoholic 

patients, often poorly compliant with therapy and therefore at 

risk for resistance if treated by direct antivirals only. VP & CH: 

equal contribution. 

Disclosures: 

The following authors have nothing to disclose: Veronique Pene, Céline Hernandez, 

Etienne Blanc, Lynda Aoudjehane, Béatrice Le Grand, Arnaud Carpentier, 

Jean-François Méritet, Filomena Conti, Yvon Calmus, Hélène Rouach, Philippe 

Podevin, Arielle R. Rosenberg 

followed by 15 weeks of combined therapy with 250mg REP 

2139-Ca and 180ug Pegasys® and then 33 weeks with 

Pegasys® monotherapy. Viremia (HDV RNA and HBV DNA), 

HBsAg and anti-HBs are followed every two weeks (Robogene 

RT-PCR, Abbott RealTime HBV, Abbott Architect) performed at 

the Institute of Virology, University of Duisburg-Essen (Essen, 

Germany). HDV RNA is validated on separate test platforms 

at the National Genetics Institute (Los Angeles, USA) and the 

Institute of Virology, Technische Universität München (Munich, 

Germany). Results: REP 2139-Ca treatment is well tolerated 

with mild and quickly resolving IV infusion reactions. Serum 

HBsAg is currently reduced 1-6 log in 11/12 patients (5 with 

serum HBsAg < 1 IU / ml) and HDV RNA is currently reduced 

1.5-7 log in 12 /12 patients (undetectable in 6 patients). Anti- 

HBs is detected in 10/12 patients, with 6 patients < 10mIU / 

ml and after combined exposure to Pegasys®, 5 patients have 

substantially increased anti-HBs titers from 50 to > 800 mIU / 

ml. In all patients with pre-treatment HBV DNA < 10 IU / ml, 

de-repression of serum HBV DNA is observed. Conclusions: 

Updated interim data from the REP 301 protocol assessing the 

safety and antiviral efficacy of REP 2139 (first in monotherapy 

and then with add on Pegasys® at week 16) in 12 Caucasian 

patients with chronic HBV / HDV co-infection demonstrated 

substantial reductions in serum HBsAg and HDV RNA as well 

as appearance of anti-HBs. HDV RNA reductions appear stronger 

than HBsAg reductions, suggesting an additional antiviral 

mechanism other than the inhibition of subviral particle assembly 

may affect HDV directly. REP 2139-Ca may become an 

important new therapeutic option for patients with chronic HBV 

/ HDV infection. 

Disclosures: 

Michel Bazinet - Board Membership: REPLICor Inc.; Employment: REPLICor Inc.; 

Management Position: REPLICor Inc.; Patent Held/Filed: REPLICor Inc.; Stock 

Shareholder: REPLICor Inc. 

Andrew Vaillant - Employment: REPLICor; Stock Shareholder: REPLICor 

The following authors have nothing to disclose: Victor Pantea, Valentin 

Cebotarescu, Lilia Cojuhari, Paulina Jimbei, Jeffrey Albrecht, Peter Schmid, Hadi 

Karimzadeh, Michael Roggendorf 

31 

Update on the safety and efficacy of REP 2139 monotherapy 

and subsequent combination therapy with 

pegylated interferon alpha-2a in chronic HBV / HDV 

co-infection in Caucasian patients 

Michel Bazinet 1 , Victor Pantea 2 , Valentin Cebotarescu 2 , Lilia 

Cojuhari 3 , Paulina Jimbei 3 , Jeffrey Albrecht 4 , Peter Schmid 4 , Hadi 

Karimzadeh 5 , Michael Roggendorf 5 , Andrew Vaillant 1 ; 1 Replicor 

Inc., Montreal, QC, Canada; 2 N. Testemitanu State University of 

Medicine and Pharmacy, Chisinau, Moldova (the Republic of); 

3 Toma Ciorba Infectious Clinical Hospital, Chisinau, Moldova 

(the Republic of); 4 National Genetics Institute, Los Angeles, CA; 

5 Institure of Virololgy, Technische Universität München, Munich, 

Germany 

Introduction: HBV / HDV co-infection causes rapid progression 

of liver disease and with no approved therapy, presents a 

significant unmet medical need. Nucleic acid polymers (NAPs) 

block HBV subviral particle assembly and release from infected 

hepatocytes and can eliminate serum HBsAg. As the NAP REP 

2139 was previously been shown to clear serum HBsAg and 

improve the ability of immunotherapy to elicit SVR in Asian 

patients with HBV, its activity in combination with Pegasys® in 

HBV / HDV co-infected Caucasian patients is currently being 

examined. Methods: In a phase II proof of concept trial (REP 

301; NCT02233075), patients with chronic HBV / HDV co-infection 

received once weekly dosing of 500mg REP 2139-Ca 

(calcium chelate complex) by 2h IV infusion for 15 weeks, 

32 

Reductions in cccDNA under NUC and ARC-520 therapy 

in chimpanzees with chronic hepatitis B virus infection 

implicate integrated DNA in maintaining circulating 

HBsAg 

Christine I. Wooddell 1 , Deborah Chavez 2 , Jason E. Goetzmann 

3 , Bernadette Guerra 2 , Ryan M. Peterson 1 , Helen Lee 2 , 

Julia O. Hegge 1 , Robert Gish 4 , Stephen Locarnini 5 , Christopher 

R. Anzalone 1 , Robert E. Lanford 2 , David L. Lewis 1 ; 1 Arrowhead 

Madison, Arrowhead Research Corporation, Madison, WI; 2 Texas 

Biomedical Research Institute, San Antonio, TX; 3 New Iberia 

Research Center, University of Louisiana at Lafayette, New Iberia, 

LA; 4 Department of Medicine, Stanford University Medical Center, 

San Diego, CA; 5 Victorian Infectious Diseases Reference Laboratory, 

Melbourne, VIC, Australia 

Background: RNAi therapeutic ARC-520 designed to target 

all cccDNA-derived transcripts reduces viral antigenemia for 

>1 month after single doses in HBV patients. Here we report 

the effect of multiple ARC-520 doses on hepatic HBV DNA 

and RNA in HBV chimps. Methods: 9 chimps (5 M, 4 F; 9-37 

yrs) received 6-11 monthly injections of ARC-520 concurrent 

with NUC therapy. 5 were HBeAg-positive (HBeAg+), baseline 

DNA 8-9 log 10 

IU/mL serum, and 4 were HBeAg-negative 

(HBeAg-), ≤3 log 10 

IU/mL. Chimps received NUCs for 

8-24 weeks prior to ARC-520 dosing. Liver biopsies from 8 

chimps were taken at baseline, completion of NUC lead-in 

and on study. HBV DNA, +/- plasmid-safe DNase digestion to


enrich cccDNA, was measured by qPCR. Pre-core/core RNA 

(C probe) and total HBV RNA (Total probe) were measured 

by RT-qPCR. The Guide for the Care and Use of Laboratory 

Animals was strictly adhered to. Results: During NUC lead-in, 

total liver HBV DNA decreased 1.1-2.5 log 10 

in HBeAg+ but 

not appreciably in HBeAg- chimps. cccDNA in HBeAg+ chimps 

decreased 0.7 ± 0.6 log 10 

. Following addition of ARC-520 in 

HBeAg+, total liver DNA decreased from baseline by 1.5–2.9 

log 10 

and cccDNA by 1.4 ± 0.7 log 10 

, the degree of reduction 

generally correlating with duration of treatment. Neither 

total HBV DNA nor cccDNA levels changed remarkably in 

HBeAg- during the study, which at baseline had 2-4 orders of 

magnitude less cccDNA than HBeAg+ chimps. HBeAg- chimps 

had 50-fold more DNase-sensitive HBV DNA, possibly indicating 

the majority is integrated DNA rather than cccDNA. 

HBV RNA was not reduced by NUCs, but with addition of 

ARC-520 RNA reductions tracked qHBsAg reductions. In 

HBeAg+, Total probe detected 1-2x as many transcripts as 

the C probe, suggesting similar levels of core/pre-core and 

S transcripts. In HBeAg-, the Total probe detected 37x more 

transcripts than the C probe, supporting a greater proportion of 

HBsAg transcripts being produced from integrated HBV DNA 

in HBeAg- chimps. Integration between DR1 and DR2 would 

result in HBV RNA lacking ARC-520 target sites, consistent 

with greater HBsAg reduction in HBeAg+ (1.7 ± 0.5 log 10 

) 

than HBeAg- chimps (0.7 ± 0.1 log 10 

). Administration of siRNA 

targeted to integrant-produced transcripts resulted in HBsAg 

reductions up to 2.3 log 10 

beyond those produced by ARC-520 

in HBeAg-. Conclusions: 1) ARC-520 reduced total liver DNA 

and cccDNA beyond levels achieved in HBeAg+ with NUCs 

during lead-in; 2) ARC-520 but not NUCs reduced HBV RNA 

and antigens; 3) integrated HBV DNA may be important in 

maintaining HBsAg in chronic HBV, especially in HBeAg-. This 

finding, if confirmed, has important implications for development 

of new HBV therapies. 

Disclosures: 

Christine I. Wooddell - Employment: Arrowhead Research Corporation 

Ryan M. Peterson - Employment: Arrowhead Research Corporation 

Julia O. Hegge - Employment: Arrowhead Research Corp 

Robert Gish - Advisory Committees or Review Panels: Gilead, AbbVie, Arrowhead; 

Consulting: Eiger, Isis, Genentech; Speaking and Teaching: Gilead, Abb- 

Vie; Stock Shareholder: Arrowhead 

Stephen Locarnini - Consulting: Gilead, Arrowhead; Employment: Melbourne 

Health 

Robert E. Lanford - Grant/Research Support: Arrowhead Research 

David L. Lewis - Employment: Arrowhead Research Corporation 

The following authors have nothing to disclose: Deborah Chavez, Jason E. Goetzmann, 

Bernadette Guerra, Helen Lee, Christopher R. Anzalone 

33 

Inhibition of Hepatitis B Virus Replication by the HBV 

Core Inhibitor NVR 3-778 

Angela Lam, Suping Ren, Robert Vogel, Christine Espiritu, Mollie 

Kelly, Vincent Lau, George D. Hartman, Lalo Flores, Klaus Klumpp; 

Novira Therapeutics Inc., Doyleston, PA 

Background: Hepatitis B virus (HBV) chronically infects more 

than 350 million people worldwide, and is a major cause of 

severe liver disease including cirrhosis and hepatocellular carcinoma. 

NVR 3-778 represents a new class of HBV core inhibitors 

and is currently in clinical development for the treatment of 

chronic hepatitis B. We present the preclinical characterization 

of NVR 3-778 and its antiviral properties in cells expressing 

HBV. Methods: HBV core assembly was examined using fluorescence 

quenching and electron microscopy. HBV replication 

inhibition was measured as reduction of secreted HBV DNA in 

HBV producing HepG2.2.15 cells. The production, encapsidation 

and secretion of HBV RNA was determined using Northern 

blot and Quantigene assays. Antiviral activity of NVR 3-778 

against nucleoside resistant HBV variants was determined by 

cell based phenotyping. HBV broad spectrum antiviral activity 

was examined by testing NVR 3-778 across all HBV genotypes 

(A-H). Results: NVR 3-778 inhibited HBV replication in 

HepG2.2.15 cells and could fully block the production of HBV 

DNA.. The compound could effectively induce mis-assembly of 

recombinant HBV core protein and electron microscopy indicated 

the formation of capsid-like particles. In HBV expressing 

cells, NVR 3-778 blocked the encapsidation of HBV pgRNA, 

HBV DNA formation, and the release of HBV DNA and RNA 

containing particles. NVR 3-778 was similarly active against 

wild-type and HBV nucleos(t)ide resistant variants with defined 

amino acid changes within the reverse transcriptase protein: 

rtL180M/M204V, rtN236T, rtA181V, rtA181V/N236T, and 

rtL180M/M204V/N236T. Furthermore, in transfection assays 

NVR 3-778 inhibited HBV replication across representative 

HBV strains from genotypes A to H (EC 50 

values from 0.20 to 

0.58 mM). Conclusions: Preclinical antiviral profiling studies 

showed that the antiviral HBV replication inhibitor NVR 3-778 

could induce mis-assembly of core protein into capsid-like particles 

in vitro. Treatment of HBV producing cells with NVR 3-778 

was associated with inhibition of pgRNA encapsidation. Consequently, 

the downstream processes of HBV DNA production 

and secretion of infectious HBV DNA-containing virions (Dane 

particles) were inhibited. In addition, NVR 3-778 also blocked 

the secretion of HBV RNA-containing particles. NVR 3-778 was 

active against HBV variants resistant to nucleos(t)ide analogs 

and was broadly active across all HBV genotypes. These results 

support the clinical development of NVR 3-778 as a potential 

new therapy for chronic hepatitis B patients alone or in combination 

with nucleoside analogs or other antiviral agents. 

Disclosures: 

Angela Lam - Employment: Novira Therapeutics, Merck & Co 

Christine Espiritu - Employment: Novira Therapeutics 

Mollie Kelly - Independent Contractor: Novira Therapeutics 

George D. Hartman - Management Position: Novira Therapeutics 

Klaus Klumpp - Board Membership: Riboscience LLC; Employment: Novira Therapeutics 

Inc 

The following authors have nothing to disclose: Suping Ren, Robert Vogel, Vincent 

Lau, Lalo Flores 

34 

Dual-gRNAs and gRNA-microRNA (miRNA)-gRNA ternary 

cassette combined CRISPR/Cas9 system and RNAi 

approach promotes the clearance of HBV cccDNA 

Jie Wang, Fengmin Lu; Peking University Health Science Center, 

Beijing, China 

Purpose: To investigate the potential use of CRISPR/Cas9 

approach for eradicating hepatitis B virus (HBV) infection, we 

designed and screened for the most effective gRNAs against 

HBV of genotypes A-D, and developed a novel gRNA-microRNA 

(miRNA)-gRNA ternary cassette. Methods: A total of 

15 gRNAs against HBV of genotypes A-D were designed. 11 

combinations of two above gRNAs (dual-gRNAs) covering the 

different regions of HBV genome were chosen. Based on the 

efficiency of dual-gRNAs in suppressing HBV replication, a 

novel gRNA-miRNA-gRNA ternary cassette driven by a single 

U6 promoter was designed by integrating an anti-HBV primiR31 

mimic between two HBV-specific gRNAs, in which two 

gRNAs could be released through Drosha/DGCR8 processing. 

The HBV replication was examined by the measurement 

of HBV surface antigen (HBsAg) and e antigen (HBeAg) in 

the culture supernatant. The destruction of HBV-expressing vec-


tor was examined by polymerase chain reaction (PCR) and 

sequencing method, and the destruction of cccDNA was examined 

by KCl precipitation, plasmid-safe ATP-dependent DNase 

digestion, rolling circle amplification and quantitative PCR combined 

method. Results: All of gRNAs could significantly reduce 

the HBsAg or HBeAg production in the culture supernatant, 

which was dependent on the region in which gRNA against. 

All of dual-gRNAs could efficiently suppress the HBsAg and 

HBeAg production for HBV of genotypes A-D, and the efficacy 

of dual-gRNAs in suppressing HBsAg and HBeAg production 

was significantly increased when compared to the single gRNA 

used alone. Furthermore, with PCR direct sequencing we confirmed 

that these dual-gRNAs could specifically destroy HBV 

expressing template by removing the fragment between the 

cleavage sites of the two used gRNAs. The gRNA-miR-HBVgRNA 

ternary cassette exerted a synergistic effect in efficiently 

inhibiting HBV replication and destroying HBV-expressing template 

in vitro and in vivo. We determined that the optimal stem 

extended length of pri-miRNA was 38 base pairs in our ternary 

cassette. Most importantly, together with RNAi approach 

the gRNA-miR-HBV-gRNA ternary cassette showed the potent 

activity on the destruction of HBV cccDNA. Conclusions: These 

results suggested that dual-gRNAs guided CRISPR/Cas9 system 

could efficiently destroy HBV expressing templates (genotypes 

A-D). Together with RNAi approach, the gRNA-miR-HBV-gRNA 

ternary cassette showed the potent activity on the destruction 

of HBV cccDNA. Since HBV cccDNA was an obstacle for the 

elimination of chronic HBV infection, thus the CRISPR/Cas9 system 

may be a potential tool for the clearance of HBV cccDNA. 

Disclosures: 

The following authors have nothing to disclose: Jie Wang, Fengmin Lu 

both GSEA and IPA. Strikingly, there was a comparable transcriptional 

response to TLR7-CM treatment in HBV-infected and 

uninfected PHH (p0.80), including a comparable 

induction of IFN-stimulated gene (ISG) signature. Notably, 

APOBEC3B (A3B) mRNA levels were only weakly induced 

by TLR7-CM (mean = 2.6-fold), and A3A expression was not 

significantly stimulated. Consistent with the lack of viral interference 

in the hepatocyte response to TLR7-CM, HBV infection 

did not result in prominent global transcriptional response in 

Mock-CM treated PHH. Knock-down of IFNAR1 mRNA (~90% 

reduction) abrogated the antiviral activity of TLR7-CM, suggesting 

that one or more of the ISGs strongly induced by treatment 

(e.g. MX1, IFI16, IFIT1 and IFITM1) may be HBV restriction 

factors. Conclusion: These data demonstrate that type I IFN is a 

key antiviral cytokine induced by GS-9620 in PBMCs, and that 

HBV does not interfere with IFN-signaling in human hepatocytes. 

In addition, transcriptional profiling identified candidate 

HBV restriction factors, but indicated that APOBEC3A likely 

does not mediate the antiviral activity of TLR7-CM in HBV-infected 

hepatocytes. 

Disclosures: 

Li Li - Employment: Gilead Sciences 

Congrong Niu - Employment: Gilead Science 

Stephane Daffis - Employment: Gilead Sciences 

Hilario Ramos - Employment: Gilead Sciences 

Eduardo Salas - Employment: Gilead Sciences; Stock Shareholder: Gilead Sciences 

Christian Voitenleitner - Employment: Gilead Sciences 

William E. Delaney - Employment: Gilead Sciences; Patent Held/Filed: Gilead 

Sciences; Stock Shareholder: Gilead Sciences 

Simon P. Fletcher - Employment: Gilead Sciences; Stock Shareholder: Gilead 

Sciences 

35 

HBV Does Not Modulate the Transcriptional Response 

to TLR7-Induced Cytokines in Highly Infected Primary 

Human Hepatocytes 

Li Li, Congrong Niu, Stephane Daffis, Hilario Ramos, Eduardo 

Salas, Christian Voitenleitner, William E. Delaney, Simon P. 

Fletcher; Gilead Sciences, Foster City, CA 

Background & Aims: GS-9620, an oral agonist of toll-like 

receptor 7 (TLR7), is in phase 2 trials for the treatment of 

chronic hepatitis B (CHB). We previously demonstrated that 

prolonged exposure to antiviral cytokines directly induced by 

TLR7 activation, such as interferon-α (IFN-α), potently inhibited 

HBV in primary human hepatocytes (PHH) in vitro. Here we 

characterized the transcriptional response of PHH to TLR7-induced 

cytokines in order to identify potential HBV restriction 

factors and to determine whether HBV infection modulates 

signaling of these cytokines in hepatocytes. Methods: PHH 

from two different donors were infected with HBV or mock 

infected, and 13 days later were treated with media from 

healthy human PBMCs stimulated with either GS-9620 (TLR7 

conditioned media; TLR7-CM) or DMSO (control; Mock-CM). 

High level infection (≥75% PHH infected) was confirmed by 

HBV core staining. Whole transcriptome analysis of PHH 

± HBV at 8, 24 and 48 hours post-treatment with TLR7-CM 

and Mock-CM was performed by RNA-Seq. Transcriptional 

responses were characterized by gene set enrichment analysis 

(GSEA) and Ingenuity Pathway Analysis (IPA). Knock-down of 

IFN-α/β receptor 1 (IFNAR1) mRNA was performed by siRNA, 

and included a negative (scramble) control. Results: TLR7-CM 

induced a comparable transcriptional signature in PHH from 

two independent donors. In both donors, TLR7-CM induced the 

greatest transcriptional response at 8 hours post-treatment, with 

IFN signaling pathways being the top induced pathways by 

36 

GalNAc-siRNA conjugate ALN-HBV targets a highly 

conserved, pan-genotypic X-orf viral site and mediates 

profound and durable HBsAg silencing in vitro and in 

vivo 

Laura Sepp-Lorenzino, Andrew G. Sprague, Tara Mayo, Tuyen 

M. Nguyen, Svetlana Shulga Morskaya, Huilei Xu, Stuart Milstein, 

Greg Hinkle, Pia Kasperkovitz, Richard G. Duncan, Natalie Keirstead, 

Brenda Carito, Lauren Moran, Prasoon Chaturvedi, Krishna 

C. Aluri, Husain Attarwala, Renta M. Hutabarat, Ju Liu, Chris Tran, 

Qianfan Wang, Benjamin S. Brigham, Akin Akinc, Klaus B. Charisse, 

Vasant Jadhav, Satya Kuchimanchi, Martin A. Maier, Muthiah 

Manoharan, Rachel Meyers, Tadeusz Wyrzykiewicz, Haroon 

Hashmi, Julie Donovan, Tim Mooney, Daniel Freedman, Tanya 

P. Sengupta, Karin Galil, Eoin Coakley, Patrick Haslett; Alnylam 

Pharmaceuticals, Cambridge, MA 

Background: Despite the prevalence of chronic HBV infection 

(CHB), there are no highly effective therapies allowing sustained 

off-treatment viral control and/or cure of HBV infection. 

An RNAi therapeutic targeting the HBV genome has the potential 

to achieve a “functional cure” by effectively decreasing 

expression of all viral gene products, including tolerogenic viral 

antigens, e.g., HBsAg. Methods and Results: ALN-HBV is a 

hepatocyte-targeted GalNAc-siRNA conjugate with Enhanced 

Stabilization Chemistry (ESC) for subcutaneous (SC) administration. 

It targets a site in the HBV X protein open reading 

frame (ORF), thereby targeting all four HBV RNA transcripts 

for degradation by RNA interference. Bioinformatic analyses 

indicate a >95% perfect sequence homology with a database 

of 3,950 full length HBV viral genomes incorporating genotypes 

A through J, and a low potential for off-target activity 

against host genes. Pan-genotypic activity was confirmed in


vitro, using reporter constructs representative of genotypes A-J. 

In HepG2.2.15 cells, ALN-HBV mediates viral transcript silencing 

with ED 50 

s of 12-19 pM, as determined for amplicons in 

the P and S ORFs. In vivo, in an AAV-HBV mouse model, a 

single 3 mg/kg SC dose of ANL-HBV resulted in mean 1.6 

log 10 

reduction in plasma HBsAg (3.6 log 10 

max reduction) at 

10-15 days post dosing. After 3 weekly doses of 3 mg/kg SC, 

mean HBsAg silencing >2.9 log 10 

was observed, with several 

animals being below the level of quantitation (100 days after the last dose. 

In vitro metabolic stability, and mouse and rat pharmacokinetics 

and liver exposure were consistent with ESC chemistry 

which enables the long-lived pharmacologic response. Exploratory 

toxicology and histopathology were performed in rats 

sacrificed 24 hr after 3 weekly doses of 30 and 100 mg/kg 

SC, and revealed excellent tolerability. Additional pharmacology 

studies are in progress, including combination studies 

with ALN-PDL, an RNAi drug targeting PD-L1 in hepatocytes. 

ALN-PDL is expected to result in augmentation of HBV-specific 

cellular immunity in the liver, thereby improving immunological 

control of HBV infection locally, while avoiding the immunotoxicity 

of systemic immune checkpoint blockade. Conclusion: 

ALN-HBV is a single, hepatocyte-targeted siRNA conjugate 

targeting a highly conserved region in the HBV genome. It 

mediates specific, potent and durable silencing of HBV viral 

transcripts and HBsAg expression. ALN-HBV is being developed 

as finite treatment in combination with standard-of-care 

nucleos(t)ide analogs as a means for inducing a functional cure 

in CHB patients. A CTA filing is planned for late 2015. 

Disclosures: 

Laura Sepp-Lorenzino - Employment: Alnylam 

Andrew G. Sprague - Employment: Alnylam Pharmaceuticals; Stock Shareholder: 

Alnylam Pharmaceuticals 

Tuyen M. Nguyen - Employment: Alnylam Pharmaceutical; Stock Shareholder: 

Alnylam Pharmaceutical 

Svetlana Shulga Morskaya - Employment: Alnylam Pharmaceuticals; Stock Shareholder: 


Stuart Milstein - Employment: Alnylam 

Greg Hinkle - Employment: Alnylam Pharmaceuticals 

Richard G. Duncan - Employment: Alnylam Pharmaceuticals; Stock Shareholder: 


Krishna C. Aluri - Employment: Alnylam Pharmaceuticals 

Husain Attarwala - Employment: Alnylam Pharmaceuticals 

Renta M. Hutabarat - Employment: Alnylam Pharmaceuticals 

Ju Liu - Employment: Alnylam 

Benjamin S. Brigham - Employment: Alnylam Pharmaceuticals 

Akin Akinc - Employment: Alnylam Pharmaceuticals 

Klaus B. Charisse - Employment: Alnylam Pharmaceuticals 

Vasant Jadhav - Employment: Alnylam Pharmaceuticals 

Satya Kuchimanchi - Employment: Alnylam Pharmaceuticals 

Martin A. Maier - Employment: Alnylam Pharmaceuticals; Stock Shareholder: 


Muthiah Manoharan - Employment: Alnylam 

Rachel Meyers - Employment: Alnylam Pharmaceuticals 

Haroon Hashmi - Employment: Alnylam Pharmaceuticals; Stock Shareholder: 


Daniel Freedman - Employment: Alnylam Pharmaceuticals 

Tanya P. Sengupta - Employment: Alnylam Pharmaceuticals 

Karin Galil - Consulting: Alnylam Pharmaceuticals 

Eoin Coakley - Employment: AbbVie; Stock Shareholder: AbbVie 

Patrick Haslett - Employment: Alnylam Pharmaceuticals 

The following authors have nothing to disclose: Tara Mayo, Huilei Xu, Pia 

Kasperkovitz, Natalie Keirstead, Brenda Carito, Lauren Moran, Prasoon 

Chaturvedi, Chris Tran, Qianfan Wang, Tadeusz Wyrzykiewicz, Julie Donovan, 

Tim Mooney 

37 

Safety and efficacy of daclatasvir plus sofosbuvir with 

or without ribavirin for the treatment of chronic HCV 

genotype 3 infection: Interim results of a multicenter 

European compassionate use program 

Tania M. Welzel 1 , Joerg Petersen 2 , Peter Ferenci 3 , Michael 

Gschwantler 4 , Kerstin Herzer 5 , Markus Cornberg 6 , Eckart Schott 7 , 

Thomas Berg 8 , Ulrich Spengler 9 , Ola Weiland 10 , Marc van der 

Valk 11 , Andreas Geier 12 , Jürgen K. Rockstroh 9 , Markus Peck-Radosavljevic 

3 , Yue Zhao 13 , Maria Jesus Jimenez Exposito 14 , Stefan 

Zeuzem 1 ; 1 Universitätsklinikum der Johann Wolfgang Goethe 

Universität, Frankfurt, Germany; 2 IFI Institut für Interdisziplinäre 

Medizin, Hamburg, Germany; 3 Medizinische Universität Wien, 

Vienna, Austria; 4 Wilhelminenspital, Vienna, Austria; 5 Universitätsklinikum 

Essen (AöR), Essen, Germany; 6 Medizinische 

Hochschule Hannover, Hannover, Germany; 7 Charité Universitätmedizin 

Berlin, Berlin, Germany; 8 Universitätsklinikum Leipzig, 

Leipzig, Germany; 9 Universitätsklinikum Bonn, Bonn, Germany; 

10 Karolinska Institutet, Solna, Sweden; 11 Academic Medical Center, 

Amsterdam, Netherlands; 12 Universitätsklinikum Würzburg, 

Würzburg, Germany; 13 Bristol-Myers Squibb, Hopewell, NJ; 

14 Bristol-Myers Squibb, Princeton, NJ 

BACKGROUND: Despite progress in the development of well 

tolerated, all-oral, interferon-free therapies, the treatment of 

HCV genotype (GT) 3 infection remains challenging. Current 

treatment options are limited, and debate continues regarding 

the optimal regimen and duration of treatment, especially for 

treatment-experienced patients with liver cirrhosis. Here we 

report interim results on the combination daclatasvir (DCV) plus 

sofosbuvir (SOF) with or without ribavirin (RBV) for the treatment 

of GT3-infected patients from a European compassionate use 

program (CUP). METHODS: This CUP enrolled adult patients 

with chronic HCV infection who were at a high risk of hepatic 

decompensation or death within 12 months if left untreated, 

and who had no available treatment options. Patients received 

DCV 60mg + SOF 400mg QD, with RBV added at the physician’s 

discretion. Recommended duration of treatment was 

24 weeks. This interim analysis includes 101 GT 3-infected 

patients enrolled in the CUP with available data on March 16, 

2015, of whom 24 patients have available SVR12 data to 

date. RESULTS: Most patients were male (68%), white (90%); 

median age: 54 years (31–75). Fifty-nine (58%) were treatment 

experienced. Most patients had cirrhosis (81%); 46 of them 

(56%) were Child-Pugh class B/C; and 13 (16%) had MELD 

scores ≥15. Seven patients were liver transplant recipients; 15 

were HIV/HCV coinfected. Median baseline HCV RNA was 

5.6 log 10 

IU/mL. Overall, SVR12 rates among GT3-infected 

patients with HCV RNA available data were 92% (22/24). 

One patient meets criteria for relapse. SVR12 rates among 

sub-groups are presented in the Table. Updated SVR12 data 

will be presented. Twenty-two patients experienced at least one 

serious adverse event (AE); 5 patients discontinued therapy due 

to AEs (pneumonia/multi-organ failure, physical deterioration, 

hepatic encephalopathy, syncope, undetermined); 2 patients 

died (multi-organ failure, liver related). CONCLUSION: In this 

preliminary analysis, DCV in combination with SOF±RBV led 

to high SVR12 rates in HCV GT3-infected patients, regardless 

of cirrhosis status, and was well tolerated. The use of RBV did 

not affect efficacy outcomes.


†All liver transplant patients had cirrhosis, ‡Includes 3 liver transplant 

patients 

Disclosures: 

Tania M. Welzel - Advisory Committees or Review Panels: Novartis, Janssen, 

Gilead, Abbvie, Boehringer-Ingelheim+, BMS 

Joerg Petersen - Advisory Committees or Review Panels: Bristol-Myers Squibb, 

Gilead, Novartis, Merck, Bristol-Myers Squibb, Gilead, Novartis, Merck; Grant/ 

Research Support: Roche, GlaxoSmithKline, Roche, GlaxoSmithKline; Speaking 

and Teaching: Abbott, Tibotec, Merck, Abbott, Tibotec, Merck 

Peter Ferenci - Advisory Committees or Review Panels: Idenix, Gilead, MSD, 

Janssen, Salix, AbbVie, BMS; Patent Held/Filed: Madaus Rottapharm; Speaking 

and Teaching: Gilead, Roche 

Michael Gschwantler - Advisory Committees or Review Panels: Janssen, BMS, 

Gilead, AbbVie; Speaking and Teaching: Janssen, BMS, Gilead, AbbVie 

Markus Cornberg - Advisory Committees or Review Panels: Merck (MSD Germamny), 

Roche, Gilead, Novartis, Abbvie, Janssen Cilag, BMS; Grant/Research 

Support: Merck (MSD Germamny), Roche; Speaking and Teaching: Merck (MSD 

Germamny), Roche, Gilead, BMS, Novartis, Falk, Abbvie 

Eckart Schott - Advisory Committees or Review Panels: Gilead, Roche, Bayer, 

BMS, Abbvie; Speaking and Teaching: Gilead, Novartis, Roche, MSD, Bayer, 

Falk, BMS, Janssen, Abbvie 

Thomas Berg - Advisory Committees or Review Panels: Gilead, BMS, Roche, 

Tibotec, Vertex, Jannsen, Novartis, Abbott, Merck, Abbvie; Consulting: Gilead, 

BMS, Roche, Tibotec; Vertex, Janssen; Grant/Research Support: Gilead, BMS, 

Roche, Tibotec; Vertex, Jannssen, Merck/MSD, Boehringer Ingelheim, Novartis, 

Abbvie; Speaking and Teaching: Gilead, BMS, Roche, Tibotec; Vertex, Janssen, 

Merck/MSD, Novartis, Merck, Bayer, Abbvie 

Ola Weiland - Advisory Committees or Review Panels: Merk, BMS, Medivir, Gilead, 

AbbVie; Grant/Research Support; Speaking and Teaching: Merk, Roche, 

BMS, Novartis, Janssen, Medivir, Gilead, AbbVie 

Marc van der Valk - Advisory Committees or Review Panels: gilead, msd, bms, 

abbvie, janssen cilag, viiV, roche 

Andreas Geier - Advisory Committees or Review Panels: AbbVie, Janssen; Speaking 

and Teaching: BMS, Gilead, Sequana, Falk, Novartis 

Jürgen K. Rockstroh - Advisory Committees or Review Panels: Abbvie, BI, BMS, 

Merck, Roche, Tibotec, Abbvie, Bionor, Tobira, ViiV, Gilead, Janssen; Consulting: 

Novartis; Grant/Research Support: Merck; Speaking and Teaching: Abbott, 

BI, BMS, Merck, Roche, Tibotec, Gilead, Janssen, ViiV 

Markus Peck-Radosavljevic - Advisory Committees or Review Panels: Bayer, Gilead, 

Janssen, BMS, AbbVie; Consulting: Bayer, Boehringer-Ingelheim, Jennerex, 

Eli Lilly, AbbVie; Grant/Research Support: Bayer, Roche, Gilead, MSD, AbbVie; 

Speaking and Teaching: Bayer, Roche, Gilead, MSD, Eli Lilly, AbbVie, Bayer 

Maria Jesus Jimenez Exposito - Employment: Bristol-Myers Squibb 

Stefan Zeuzem - Consulting: Abbvie, Bristol-Myers Squibb Co., Gilead, Merck 

& Co., Janssen 

The following authors have nothing to disclose: Kerstin Herzer, Ulrich Spengler, 

Yue Zhao 

38 

Sofosbuvir/GS-5816+GS-9857 for 6 or 8 Weeks in 

Genotype 1 or 3 HCV-infected Patients 

Edward J. Gane 1 , Robert H. Hyland 2 , Yin Yang 2 , Luisa M. Stamm 2 , 

Diana M. Brainard 2 , John G. McHutchison 2 , Catherine A. Stedman 

3 ; 1 Auckland Clinical Studies, Auckland, New Zealand; 2 Gilead 

Sciences, Inc, Foster City, CA; 3 Christchurch Clinical Studies 

Trust, Christchurch, New Zealand 

Background: Sofosbuvir (SOF), GS-5816, and GS-9857 target 

3 distinct viral proteins, NS5B, NS5A, and NS3, respectively. 

The combination of these pangenotypic agents has the potential 

to improve efficacy and may reduce treatment duration 

across different patient populations. We previously reported 

safety and efficacy of SOF/GS-5816 + GS-9857 administered 

for 4 or 6 weeks in patients with HCV genotype 1 (GT1) infection, 

including treatment-naïve (TN) GT1 patients without cirrhosis; 

TN GT1 patients with cirrhosis; and GT1 patients who had 

failed a regimen containing at least 2 direct antiviral agents. 

We now evaluate the safety and efficacy of this regimen 

administered for 6 or 8 weeks in more difficult-to-treat populations, 

including treatment experienced (TE) GT 1 patients with 

cirrhosis who had failed PEG/RBV; TE GT 1 patients who had 

failed protease inhibitor (PI) + PEG/RBV; TN GT 3 patients with 

cirrhosis; and TE GT 3 patients with cirrhosis who had failed 

PEG/RBV. Methods: All patients received once daily SOF/ 

GS-5816 400mg/100mg in a fixed dose combination, plus 

GS-9857 100mg. Treatment duration of each group differed 

according to the patient’s baseline characteristics: 8 weeks 

for TE GT 1 patients who had failed prior PEG/RBV, 8 weeks 

for TE GT 1 patients who had failed PI + PEG/RBV, 6 weeks 

for TN GT 3 patients and 8 weeks for TE GT 3 patients. The 

primary efficacy endpoint was SVR12, defined as HCV RNA 


39 

SVR12 results from the Phase II, open-label IMPACT 

study of simeprevir (SMV) in combination with daclatasvir 

(DCV) and sofosbuvir (SOF) in treatment-naïve and 

-experienced patients with chronic HCV genotype 1/4 

infection and decompensated liver disease 

Eric Lawitz 1 , Fred Poordad 1 , Julio A. Gutierrez 1 , Thomas Kakuda 2 , 

Gaston Picchio 3 , Greet Beets 4 , Ann Vandevoorde 4 , Peter Van 

Remoortere 2 , Bert Jacquemyn 4 , Gemma Quinn 4 , Donghan Luo 2 , 

Sivi Ouwerkerk-Mahadevan 5 , Leen Vijgen 4 , Veerle Van Eygen 4 , 

Maria Beumont 5 ; 1 Texas Liver Institute, University of Texas Health 

Science Center, San Antonio, TX; 2 Janssen Research & Development, 

LLC, Titusville, NJ; 3 Infectious Diseases, Janssen Research 

& Development, San Francisco, CA; 4 Janssen Infectious Diseases 

BVBA, Beerse, Belgium; 5 Janssen Research & Development, 

Beerse, Belgium 

Purpose: The ongoing IMPACT study (NCT02262728) 

assessed SMV+DCV+SOF in HCV genotype (GT)1/4-infected 

patients (pts) with decompensated liver disease. We present 

sustained virologic response 12 weeks (w) after end of treatment 

(SVR12) from an interim analysis (IA). Methods: Treatment-naïve 

or PegIFN±RBV treatment-experienced cirrhotic 

adults with Child-Pugh (CP) score


patients ± cirrhosis ± HIV co-infection. Injection drug use is the 

major risk factor for HCV transmission in many settings; however, 

data on interferon-free treatment outcomes among PWID 

are very limited. C-EDGE COSTAR is a Phase 3 trial evaluating 

efficacy and safety of GZR/EBR among treatment naïve 

HCV GT1/4/6-infected patients ± cirrhosis who are receiving 

opioid agonist therapy (OAT) (methadone or buprenorphine). 

METHODS: This double-blind, placebo-controlled study randomized 

patients 2:1 to an immediate treatment group (ITG) or 

a deferred treatment group (DTG). The ITG received GZR/ 

EBR for 12 weeks; the DTG received placebo for 12 weeks, 

followed by 12 weeks of open label GZR/EBR. Use of non-prescribed 

drugs was monitored by urine drug screening, but 

was not exclusionary to study participation. HCV RNA levels 

were measured using COBAS TaqMan v2.0 (lower limit of 

quantitation 5x 

ULN after normalization while on therapy. SVR12 results will 

be presented. CONCLUSION: Preliminary data indicate that 

GZR/EBR is safe and highly effective in patients with chronic 

HCV GT1, 4, or 6 receiving OAT, and supports enhanced 

efforts to address barriers to HCV treatment access for PWID. 

Disclosures: 

Gregory Dore - Board Membership: Gilead, Merck, Abbvie, Bristol-Myers 

Squibb; Grant/Research Support: Gilead, Merck, Abbvie, Bristol-Myers Squibb; 

Speaking and Teaching: Gilead, Merck, Abbvie, Bristol-Myers Squibb 

Frederick Altice - Grant/Research Support: Gileaad, NIH, NIDA, SAMHSA, 

HRSA; Speaking and Teaching: Merck, Bristol Myers Squibb, Gilead, Rush Practice 

Point Communications 

Alain H. Litwin - Advisory Committees or Review Panels: Merck, Gilead; Grant/ 

Research Support: Merck, Gilead 

Olav Dalgard - Advisory Committees or Review Panels: MSD, Janssen Cilag, 

Medivir, Gilead, Abbvie; Grant/Research Support: MSD, Medivir, Gilead 

Edward J. Gane - Advisory Committees or Review Panels: Novira, AbbVie, Janssen, 

Gilead Sciences, Janssen Cilag, Achillion, Merck, Tekmira; Speaking and 

Teaching: AbbVie, Gilead Sciences, Merck 

Anne Luetkemeyer - Grant/Research Support: Gilead, Abbvie, Pfizer, Bristol 

Myers Squibb, Merck 

Ronald Nahass - Advisory Committees or Review Panels: Gilead, MErck, Janssen, 

BMS; Grant/Research Support: Gilead, Merck, Janssen, BMS; Speaking and 

Teaching: Gilead, Merck, Janssen 

Cheng-Yuan Peng - Advisory Committees or Review Panels: AbbVie, BMS, Gilead, 

MSD, Roche 

Brian Conway - Advisory Committees or Review Panels: Vertex Pharmaceuticals, 

Merck, Boehringer Ingelheim, Jannsen Pharmaceuticals; Grant/Research Support: 

Vertex Pharmaceuticals, Merck, Boehringer Ingelheim, Jannsen Pharmaceuticals, 

AbbVie, Gilead Sciences, Gilead Sciences 

Jason Grebely - Advisory Committees or Review Panels: Merck, Gilead; Grant/ 

Research Support: Merck, Gilead, Abbvie, BMS 

Anita Y. Howe - Employment: Merck Research Laboratory 

Bach-Yen T. Nguyen - Employment: Merck 

Janice Wahl - Employment: Merck & Co, 

Eliav Barr - Employment: Merck 

Michael Robertson - Employment: Merck; Stock Shareholder: Merck 

The following authors have nothing to disclose: Oren Shibolet, Heather L. Platt 

41 

98%–100% SVR4 in HCV Genotype 1 Non-Cirrhotic 

Treatment-Naïve or Pegylated Interferon/Ribavirin Null 

Responders With the Combination of the Next Generation 

NS3/4A Protease Inhibitor ABT-493 and NS5A 

Inhibitor ABT-530 (SURVEYOR-1) 

Fred Poordad 2 , Franco Felizarta 3 , Armen Asatryan 1 , Tarek I. Hassanein 

4 , Humberto I. Aguilar 5 , Jacob P. Lalezari 6 , J. Scott Overcash 

7 , Teresa Ng 1 , Ran Liu 1 , Chih-Wei Lin 1 , Federico J. Mensa 1 , 

Jens Kort 1 ; 1 AbbVie, Inc., North Chicago, IL; 2 Texas Liver Institute, 

University of Texas Health Science Center, San Antonio, TX; 3 Private 

practice, Bakersfield, CA; 4 Southern California GI and Liver 

Centers and Southern California Education and Research Center, 

Coronado, CA; 5 Louisiana Research Center, Shreveport, LA; 

6 Quest Clinical Research, San Francisco, CA; 7 eStudySite, San 

Diego, CA 

Background: The next-generation HCV direct-acting antivirals 

(DAAs) ABT-493, an NS3/4A protease inhibitor identified by 

AbbVie and Enanta, and ABT-530, an NS5A inhibitor, are 

characterized by potent pangenotypic in vitro antiviral activity 

against major HCV genotypes (GTs), including activity against 

key known resistance-associated variants and a high barrier 

to resistance selection. Monotherapy with ABT-493 or ABT- 

530 resulted in a mean 4 log 10 

IU/mL decline from baseline 

in HCV plasma viral load in GT1-infected subjects with and 

without compensated cirrhosis. Purpose: In this phase 2 study, 

treatment with ABT-493 and ABT-530 for 12 weeks is evaluated 

in HCV GT1-infected subjects without cirrhosis. Efficacy 

and safety results are reported here. Methods: Non-cirrhotic 

GT1-infected treatment-naïve (TN) or pegylated interferon/ribavirin 

(pegIFN/RBV) null responder subjects received once-daily 

ABT-493 200 mg + ABT-530 120 or 40 mg for 12 weeks, 

and subsequently were followed for 24 weeks. The primary 

efficacy endpoint was sustained virologic response 12 wks 

after the last dose of study drug (SVR12). Safety was evaluated 

by adverse event (AE) monitoring, laboratory testing, 

and other standard assessments. Results: 79 subjects (male, 

52%; median [range] age, 54.0 [26.0–70.0] years; GT1a, 

81%; GT1b, 19%; TN, 63%; pegIFN/RBV null responders, 

37%; fibrosis >F2, 25%; median [range] HCV RNA log 10 

IU/ 

mL, 6.8 [4.4–7.5]) were enrolled, 40 received ABT-493 200 

mg+ABT-530 120 mg and 39 received ABT-493 200 mg and 

ABT-530 40 mg. SVR 4 weeks after the last dose of study drug 

(SVR4) is available in all 79 subjects. SVR4 was achieved in 

29 of 29 (100%) pegIFN/RBV null responders and 49 of 50 

(98%) TN subjects. One TN subject relapsed at post-treatment 

week 4; further characterization of this relapse and complete 

post-treatment week 12 data (SVR12) will be available. There 

were no treatment-related serious AEs or clinically relevant 

laboratory findings. The most common AEs (reported in >5% 

of subjects) were fatigue, headache, nausea, diarrhea, and 

anxiety. Conclusions: Once-daily 12-week treatment with the 

next-generation HCV DAAs ABT-493 and ABT-530 of GT1 

infection in non-cirrhotic TN and pegIFN/RBV null responders 

resulted in high SVR4 (98%–100%) rates. One treatment 

relapse was observed. Based on these encouraging results, 

the SURVEYOR-1 study with once-daily ABT-493 and ABT-530 

has been expanded to include GT1 subjects with compensated 

cirrhosis and evaluate a shorter, 8 week treatment duration in 

non-cirrhotic subjects.


Disclosures: 

Fred Poordad - Advisory Committees or Review Panels: Abbott/Abbvie, Achillion, 

BMS, Inhibitex, Boeheringer Ingelheim, Pfizer, Genentech, Idenix, Gilead, 

Merck, Vertex, Salix, Janssen, Novartis; Grant/Research Support: Abbvie, 

Anadys, Achillion, BMS, Boehringer Ingelheim, Genentech, Idenix, Gilead, 

Merck, Pharmassett, Vertex, Salix, Tibotec/Janssen, Novartis 

Franco Felizarta - Grant/Research Support: AbbVie, Gilead, Janssen, Merck, 

BMS, Boehringer-Ingelheim, Vertex, Roche; Speaking and Teaching: AbbVie, 

Gilead, Janssen, Merck 

Armen Asatryan - Employment: AbbVie 

Tarek I. Hassanein - Advisory Committees or Review Panels: AbbVie, Bristol-Myers 

Squibb; Grant/Research Support: AbbVie Pharmaceuticals, Obalon, Bristol-Myers 

Squibb, Eiasi Pharmaceuticals, Gilead Sciences, Janssen R&D, Idenix 

Pharmaceuticals, Ikaria Therapeutics, Merck Sharp & Dohme, NGM BioPharmaceuticals, 

Ocera Therapeutics, Salix Pharmaceuticals, Sundise, TaiGen Biotechnology, 

Takeda Pharmaceuticals, Vital Therapies, Tobria; Speaking and 

Teaching: Baxter, Bristol-Myers Squibb, Gilead, Salix, AbbVie 

Humberto I. Aguilar - Speaking and Teaching: abbvie, gilead 

Teresa Ng - Employment: AbbVie; Patent Held/Filed: AbbVie; Stock Shareholder: 

AbbVie 

Ran Liu - Employment: Abbvie 

Chih-Wei Lin - Employment: Abbvie 

Federico J. Mensa - Employment: Abbvie Inc.; Stock Shareholder: Abbvie Inc. 

Jens Kort - Employment: AbbVie Inc.; Stock Shareholder: AbbVie Inc. 

The following authors have nothing to disclose: Jacob P. Lalezari, J. Scott Overcash 

42 

An Integrated Analysis of 402 Compensated Cirrhotic 

Patients With HCV Genotype (GT) 1, 4 or 6 Infection 

Treated With Grazoprevir/Elbasvir 

Ira M. Jacobson 1 , Eric Lawitz 2 , Paul Y. Kwo 3 , Christophe Hezode 4 , 

Cheng-Yuan Peng 5 , Anita Y. Howe 6 , Peggy Hwang 6 , Janice 

Wahl 6 , Michael Robertson 6 , Eliav Barr 6 , Barbara A. Haber 6 ; 

1 Medicine, Mt. Sinai Beth Israel, New York, NY; 2 Texas Liver Institute, 

University of Texas Health Science Center, San Antonio, TX; 

3 Indiana University School of Medicine, Indianapolis, IN; 4 Henri 

Mondor Hospital, Creteil, France; 5 China Medical University Hospital, 

Taichung, Taiwan; 6 Merck & Co., Inc., Kenilworth, NJ 

Purpose: Treatment of cirrhotic HCV-infected patients (pts) often 

requires ribavirin (RBV) and long durations of therapy. In the 

C-EDGE trials, 12-16 weeks’ therapy with grazoprevir 100 

mg/elbasvir 50 mg (GZR/EBR) ± RBV was highly efficacious 

in a diverse population of pts. An analysis of the efficacy/ 

safety of GZR/EBR ± RBV among compensated cirrhotics in 

the phase 2/3 GZR/EBR program was conducted. Methods: 

Treatment-naive (TN) and treatment-experienced (TE) pts with 

compensated cirrhosis received GZR/EBR ±RBV for 12 (TN) or 

12-18 weeks (TE) in 6 studies. The primary end point was HCV 

RNA


could predict the presence of various cancer entities in patients 

with hepatocellular, colorectal and non-small cell lung carcinoma 

(HCC, CRC, NSCLC, resp.), as a tool for cancer screening 

and therapy response monitoring. Methods: MPs were 

isolated from serum of patients with HCC, CRC, NSCLC and 

from healthy donors. MP profiling was done after differential 

ultracentrifugation using FACS analysis for the presence and 

quantity of AnnexinV + /EpCAM + and AnnexinV + /EpCAM + / 

CD147 + taMPs. Results: AnnexinV + /EpCAM + and AnnexinV 

+ /EpCAM + /CD147 + taMPs highly significantly separated 

patients with carcinomas from healthy controls. The calculated 

cut-off values for AnnexinV + /EpCAM + taMPs were 36.6 

/1AnnexinV + MPs in NSCLC, 35.8/1kAnnexinV + MPs in CRC 

and 31.4/1kAnnexinV + taMPs in HCC. Additionally, AnnexinV 

+ /EpCAM + /CD147 + taMPs indicated the presence of cancer 

as well as AnnexinV + /EpCAM + taMPs alone. AnnexinV + / 

EpCAM + taMPs and AnnexinV + /EpCAM + /CD147 + taMPs 

were increased 2.4-fold (p


respectively. The median (IQR) time intervals were 46.5 (62) 

and 47.5 (87) days between MRE and biopsy and between 

ARFI and biopsy, respectively. For diagnosing any fibrosis, 

MRE had an area under ROC curve (AUROC) of 0.799 (95% 

CI, 0.723-0.875), which was significantly (p=0.012) higher 

than ARFI, which had an AUROC of 0.664 (95% CI, 0.568- 

0.760). The AUROCs of MRE for diagnosing ≥ stages 2, 3, 

and 4 fibrosis were 0.885 (95% CI, 0.816-0.953), 0.934 

(95% CI, 0.863-1.000), and 0.882 (95% CI, 0.729-1.000), 

and those for ARFI were 0.848 (95% CI, 0.776-0.921), 0.896 

(95% CI, 0.824-0.968), and 0.862 (95% CI, 0.721-1.000). 

MRE had a higher AUROC than ARFI for discriminating dichotomized 

fibrosis stages at every dichotomization cutpoint, but 

the difference in AUROC was smaller as the dichotomization 

cutpoint increased. Conclusion: In a head-to-head comparison 

and using contemporaneous liver biopsy as the gold standard, 

MRE provides significantly higher diagnostic accuracy than 

ARFI for diagnosing any fibrosis in NAFLD patients. 

Disclosures: 

Claude B. Sirlin - Grant/Research Support: GE, Pfizer, Bayer, Guerbet, Siemens 

Rohit Loomba - Advisory Committees or Review Panels: Galmed Inc, Tobira Inc, 

Arrowhead Research Inc; Consulting: Gilead Inc, Corgenix Inc, Janssen and 

Janssen Inc, Zafgen Inc, Celgene Inc, Alnylam Inc, Inanta Inc, Deutrx Inc; Grant/ 

Research Support: Daiichi Sankyo Inc, AGA, Merck Inc, Promedior Inc, Kinemed 

Inc, Immuron Inc, Adheron Inc 

The following authors have nothing to disclose: Jeffrey Y. Cui, Elhamy Heba, 

Carolyn Hernandez, William Haufe, Jonathan Hooker 

46 

The predictive value of 13 C-Methacetin breath test (MBT) 

for liver transplantation (LT) and death: a seven year 

follow-up study of 206 chronic hepatitis C patients 

Oliver Goetze 2,1 , Monika Breuer 1 , Andreas Geier 2,1 , Michael 

Fried 1 , Achim Weber 4 , Yaron Ilan 3 , Beat Mullhaupt 1 ; 1 Department 

of Gastroenterology & Hepatology, University Hospital Zurich, 

Zurich, Switzerland; 2 Division of Hepatology, University Hospital 

Würzburg, Würzburg, Germany; 3 Department of Medicine, 

Hebrew University-Hadassah Medical Center, Jerusalem, Israel; 

4 Institute of Surgical Pathology, University Hospital Zurich, Zurich, 

Switzerland 

In patients with chronic liver disease, currently used biomarkers 

demonstrate limited mid- to long-term predictive values to 

forecast deterioration of liver function leading to liver transplantation 

(LT) or liver related death. Aim: To determine the 

predictive value of the 13 C-Methacetin Breath Test (MBT) 

(BreathID ® ; Exalenz) in forseeing deterioration of liver function 

leading to LT and/or death in patients with chronic HCV infection. 

Methods: A total of 265 chronic HCV infected patients 

were enrolled, out of which 206 patients (71 females) without 

co-infections and/or liver comorbidities were considered for 

this analysis. The average BMI was 24.8 kg/m 2 (SD 4.0 kg/ 

m 2 ), 183 patients underwent liver biopsy and ISHAK scoring 

and 196 patients underwent MBT. 13 C-Methacetin peak percent 

dose recovery rate (PDR-Peak) was used as determinant 

parameter for this analysis. Patients were followed for up to 

7 years according to standard of care. During the course of 

follow-up some patients underwent antiviral treatment as part of 

routine patient management. Primary analysis was performed 

on all patients up to one of the following: liver-transplantation, 

death or response to antiviral treatment, resulting in 161 

and 158 patients with biopsy and/or MBT results respectively. 

Predictive value was assessed using ROC curve and survival 

analyses. Results: In seven years follow up, PDR-Peak, using 

cut-off of 20%/h, predicted LT and/or death with HR=12.07 

(95%CI 3.48;41.84),p


black tea. The mean ALT noted in 657 individuals within 3 

months of TE was 60.6 U/L. The average BMI was 25.7 kg/m 2 

and weight was 71.7 kg. Average daily alcohol consumption 

was 5 g/day. The average liver stiffness for the 1155 individuals 

was 8.4 kPa. CAP was available from 2013 in 719 individuals 

with average steatosis measures of 214 dB/m. After 

MLR taking into account age, alcohol, smoking and weight, 

individuals with HCV benefited from coffee intake with LSM 

on average 2 kPa less with intake of 2 or more cups of coffee 

daily (p = 0.026). This was not seen in those with HBV or 

NAFLD. In NAFLD a decrease in CAP of 23 dB/m (9%) was 

noted in individuals who drank ≥2 cups of coffee (p = 0.032). 

There was no effect of coffee consumption on CAP in other 

groups. Conclusions: Individuals with self-reported daily coffee 

intake ≥2 cups of coffee/ day had a significant decrease in 

liver stiffness, which was not seen in those with HBV or NAFLD. 

Similar daily coffee intake resulted in significant decrease in 

liver steatosis in NAFLD. These findings are consistent with the 

literature to date and add to the growing body of evidence 

suggesting coffee may be a beneficial supplement in some 

liver diseases. 

Disclosures: 

The following authors have nothing to disclose: Alexander Hodge, Sarah P. Lim, 

Evan Goh, Ophelia H. Wong, Philip Marsh, Virginia Knight, Yong Song 

48 

Serum Lipidomic Profiling for Screening Potential Biomarkers 

of Liver Cirrhosis among Patients with Chronic 

Hepatitis C 

Ana Maria Passos-Castilho 1 , Maria Lucia Ferraz 2 , Valdemir M. 

Carvalho 3 , Karina H. Cardozo 3 , Luciana Kikuchi 4 , Aline Chagas 4 , 

João Renato R. Pinho 4 , Michele S. Gomes-Gouvêa 4 , Fernanda 

Malta 4 , Flair J. Carrilho 4 , Celso Granato 1,3 ; 1 Division of Infectious 

Diseases, Federal University of Sao Paulo, Sao Paulo, Brazil; 

2 Department of Gastroenterology, Federal University of Sao Paulo, 

Sao Paulo, Brazil; 3 Fleury SA Group, Sao Paulo, Brazil; 4 Department 

of Gastroenterology, University of Sao Paulo School of Medicine, 

Sao Paulo, Brazil 

Background: Liver cirrhosis (LC) is the 14th most common cause 

of death in adults worldwide, resulting in 1.03 million deaths 

per year. Chronic hepatitis C (CHC) is one of the main causes 

of LC. Non-invasive markers of fibrosis have been increasingly 

studied as they may represent a more informative, safe and 

accessible diagnostic and/or screening tool for at-risk patients. 

Aim: To explore the serum lipidome profiles of LC to identify 

potential biomarkers. Methods: A total of 182 subjects were 

enrolled from 2011 to 2014. An ultraperformance liquid chromatography–mass 

spectrometry (UPLC-MS) lipidomic method 

was used to characterize serum profiles from LC (n=59), CHC 

(n=65) and healthy subjects (n=58). Patients were diagnosed 

by clinical, laboratory and imaging evidence of LC while 

healthy controls had normal liver function and no infectious diseases. 

Reversed-phased analysis was performed on a Waters 

ACQUITY IClass UPLC system coupled to a Waters Synapt-MS 

hidrid quadrupole-time of flight mass spectrometer operating 

in the positive ion electrospray mode with a mass scan range 

200–1200 m/z for data acquisition in continuous mode. All 

data were processed with Progenesis software (Waters). The 

resulting multivariate data set was analyzed with MetaboAnalyst 

(TMIC) using supervised partial least-squares discriminate 

analysis (PLS-DA). Potential biomarkers were selected according 

to the variable importance in the projection (VIP) and statistical 

significance was evaluated using Mann-Whitney test. The 

receiver operating characteristic (ROC) curve was performed 

to assess the accuracy of selected biomarkers in LC diagnosis. 

To identify the potential biomarkers, the HMDB and Lipid 

Maps databases were queried with the exact mass. Results: 

The UPLC–MS-based serum lipidomic profile detected 51 of 

59 LC cases, performing with a diagnostic accuracy of 87%. 

The 3 potential biomarkers differentiated LC from CHC individually 

with 76 to 88% sensitivity and 72 to 74% specificity. 

They all presented with fold change higher than 2 and p < 

0.0001 in univariate analyses. The combination of the 3 potential 

biomarkers resulted in an area under the curve of 0.95, 

86% sensitivity and 88% specificity. The model was validated 

with 1000 permutation tests (p < 0.001) to prevent overfitting. 

3-Hydroxy-cis-5-tetradecenocylcarnitine, a hydroxy fatty acid 

involved in many metabolic and proinflammatory pathways, 

exhibited a significant increase in LC and a decrease in CHC 

and was proposed as an important indicator of LC. Conclusions: 

UPLC-MS lipid profiling proved to be an efficient and 

convenient tool for diagnosis and screening of LC in an at-risk 

population. 

Disclosures: 

João Renato R. Pinho - Employment: Hospital Israelita Albert Einstein 

The following authors have nothing to disclose: Ana Maria Passos-Castilho, 

Maria Lucia Ferraz, Valdemir M. Carvalho, Karina H. Cardozo, Luciana Kikuchi, 

Aline Chagas, Michele S. Gomes-Gouvêa, Fernanda Malta, Flair J. Carrilho, 

Celso Granato 

49 

Comparison of Liver Transplant-related Survival Benefit 

in Patients With vs Without Hepatocellular Carcinoma in 

the United States. 

George N. Ioannou 1,2 , Kristin Berry 1 ; 1 Veterans Affairs Puget 

Sound Healthcare System, Seattle, WA; 2 University of Washington, 

Seattle, WA 

Background and Aims Patients with T2 hepatocellular carcinoma 

(HCC) can obtain an exception allowing them to undergo 

liver transplantation at much lower actual Model for End Stage 

Liver Disease (MELD) scores than patients without HCC. We 

aimed to compare patients transplanted with and without HCC 

with regards to transplantation-related survival benefit. Methods 

We modeled the post-transplant survival of adult, first-time 

liver transplant recipients with (n=9135) or without (n=25,890) 

HCC from 2002-2013 using Cox proportional hazards regression. 

We modeled waitlist survival of patients listed for transplantation 

with (n=15,605) or without (n=85,229) HCC using 

competing risks analysis and a combined outcome of death 

or liver failure, defined as MELD score ≥30. We used these 

survival models to calculate monthly transition probabilities and 

5-year life expectancies. Survival benefit was calculated as the 

difference between post-transplant and waitlist life expectancy. 

Results Five-year survival benefit increased with actual MELD 

score for both patients with and without HCC from just a few 

months in patients with low MELD scores (6-8) to 4 years in 

patients with the highest MELD scores (36-40). The survival 

benefit of patients with HCC was similar to that of patients 

without HCC who had the same actual MELD score, irrespective 

of tumor burden or serum alpha fetoprotein level. Because 

patients with HCC were transplanted at a lower mean MELD 

score (13.3±6.2) than patients without HCC (21.8±8.0), a 

dramatically lower mean 5-year survival benefit was achieved 

by transplanting patients with HCC (0.12 years/patient) than 

patients without HCC (1.47 years/patient). Conclusions The 

HCC MELD exception policy unintentionally resulted in a dramatic 

reduction in transplant-related survival benefit.


Five-year transplant-related survival benefit as a function of MELD 

score shown separately in patients with or without HCC 

considered, wait time 18 months (HR 1.6, 95% CI 

1.01-2.5, p=0.043) and AFP >400 ng/mL at HCC diagnosis 

(HR 3.0, 95% CI 1.7-5.5, p100 ng/mL at LT 

(HR 1.6, 95% CI 1.04-2.6, p


LT. Therefore, MHE, i) should not to be ignored and ii) deserves 

to be treated in order to reduce the risk of neurological complications 

occurring post-LT. 

Disclosures: 

The following authors have nothing to disclose: Marc-André Clément, Cristina R. 

Bosoi, Mélanie Tremblay, Chantal Bemeur, Christopher F. Rose 

52 

The Functional Basis for Cognitive Improvement after 

Liver Transplant: A Prospective Brain MRI Study 

Vishwadeep Ahluwalia, James Wade, HoChong Gilles, Melanie 

White, Ariel Unser, Edith A. Gavis, Mohammad S. Siddiqui, Velimir 

A. Luketic, Arun J. Sanyal, Puneet Puri, Douglas M. Heuman, 

Scott C. Matherly, R. Todd Stravitz, Richard K. Sterling, Michael 

Fuchs, Jasmohan S. Bajaj; VCU and McGuire VAMC, Richmond, 

VA 

Cirrhotics have varying degrees of cognitive recovery post-liver 

transplant (LT) depending on their prior hepatic encephalopathy(HE) 

status but the mechanism is unclear. Aim: Define the 

functional basis of cognitive change in LT recipients. Method: 

Cirrhotics on the transplant list underwent standard cognitive 

testing & questionnaires regarding depression (Beck Depression, 

BDI), QOL (Sickness impact profile, SIP has psych & 

physical components) pre/6 months post-LT. Cognitive tests 

were number connection A/B (NC), digit symbol (DS) & block 

design (BD) which test for psychomotor speed, set-shifting and 

visuo-motor coordination. We also performed brain functional 

MRI while subjects underwent the inhibitory control test (ICT) 

within the scanner. ICT tests for response inhibition, working 

memory & psychomotor speed. The outcomes are lures (that 

need to be inhibited) & targets (need to be responded to). 

Changes in brain activation on correct lure inhibition & target 

response were compared before & after LT. Results: We 

enrolled 67 cirrhotics of which 11 died prior, 12 still await & 

5 pts were


may help refine post-LT HCC surveillance strategies and identify 

patients who would derive the most benefits from future post-LT 

adjuvant therapies. 

RETREAT Score 

0 points for: AFP


from BM macrophages and Kupffer cells in a dose-dependent 

manner; 5-10nM MCC950 abrogated such IL-1β release. Conclusions: 

These data demonstrate that MCC950, a potent and 

selective NLRP3 inflammasome inhibitor, prevents or reverses 

inflammation, injury and fibrosis in two different murine models 

of NASH. These effects could be explained, at least in part, by 

the effective abrogation of cholesterol crystal-induced NLRP3 

activation and IL-1β release in BM macrophages and Kupffer 

cells. Targeting NLRP3 is a logical new direction in NASH 

pharmacotherapy. 

Disclosures: 

The following authors have nothing to disclose: Auvro R. Mridha, Alexander 

Wree, Avril A. Robertson, Narci C. Teoh, Matthew A. Cooper, Ariel E. Feldstein, 

Geoffrey C. Farrell 

56 

Role of Fatty Acid Desaturase 1 (FADS1) in Nonalcoholic 

Fatty Liver Disease (NAFLD) 

Shaminie Athinarayanan 1 , Yang-Yi Fan 2 , Robert Chapkin 2 , Wanqing 

Liu 1 ; 1 Medicinal Chemistry and Molecular Pharmacology, 

Purdue University, West Lafayette, IN; 2 Center for Translational 

Environmental Health Research, Texas A&M, College Station, TX 

Background: NAFLD and nonalcoholic steatohepatitis (NASH) 

are common chronic liver diseases. Previous studies have 

demonstrated that decreased hepatic and blood levels of 

n-3 polyunsaturated fatty acids (PUFA) are associated with 

NAFLD/NASH. The FADS1 gene encodes delta-5 desaturase 

(D5D), one of the rate-limiting enzymes involved in n-3 PUFA 

metabolism. We hypothesized that FADS1 is involved in the 

pathogenesis of NAFLD/NASH via modulating hepatic lipid 

homeostasis and other pathways. Methods: In order to corroborate 

our hypothesis, we used both the HepG2 cell line 

and a Fads1 knockout (null) mouse model. The FADS1 gene 

was knocked down using siRNA in HepG2 cells. Palmitic acid 

(PA)-induced lipid accumulation was compared between the KD 

cells and the control siRNA group. In addition, RNA-seq based 

hepatic transcriptome data were compared between the null 

(n=4) and wild-type (n=4) mice. Differential gene expression 

data were analyzed using EdgeR from Bioconductor. Detailed 

pathway analyses were performed by combining both a targeted 

approach to examine well-established lipid homeostasis 

pathways as well as a global, unsupervised enrichment analysis 

using Ingenuity Pathway Analysis (IPA). Results: FADS1 

knock down in HepG2 cells significantly increased PA-induced 

intracellular lipid accumulation by 60% (p < 0.001). Liver samples 

from Fads1 null mice exhibited a significantly (p


58 

Hepatocytes Release Ceramide-rich Proinflammatory 

Extracellular Vesicles in an IRE1alpha-dependent manner 

Eiji Kakazu 1,2 , Amy S. Mauer 1 , Harmeet Malhi 1 ; 1 Gastroenterology 

and Hepatology, Mayo Clinic, Rochester, MN; 2 Department of 

Community Medical Supports, Tohoku University, Aobaku, Japan 

Palmitate (PA), a lipotoxic free fatty acid, is implicated in 

hepatocyte apoptosis, macrophage-mediated liver inflammation, 

and activation of the IRE1alpha branch of the endoplasmic 

reticulum (ER) stress response - all key features of 

progressive nonalcoholic steatohepatitis (NASH). Recently, 

extracellular vesicles (EVs) have been implicated in NASH. 

However, the exact pathways of hepatocyte EV generation 

under lipotoxic conditions and their downstream effects on 

macrophages are undefined. Therefore, we hypothesized that 

hepatocytes release proinflammatory PA-induced EVs via an 

ER stress response. To test this hypothesis we employed immortalized 

mouse hepatocytes (IMH) from wild-type (WT) or IRE1a 

knockout (KO) mice, Huh 7 and primary mouse hepatocytes 

(PMH). Cells were treated with 400μM PA or thapsigargin 

(Tg) to induce ER stress. EVs were isolated from cell culture 

supernatants by ultracentrifugation and characterized by immunofluorescence 

and western blotting for EV markers, electron 

microscopy (EM) and by nanoparticle tracking analysis (NTA) 

for morphology and size. Ceramides were measured by mass 

spectrometry. CRISPR/Cas9 technology was used to delete 

XBP1. Results: PA and Tg significantly increased EV release 

in all three hepatocyte cells tested (IMH, Huh7 and PMH). The 

released EVs were 100nm in size, decorated with EV markers 

CD63, TSG101 and LAMP1, and demonstrated characteristic 

cup-shaped morphology on EM. As PA and Tg activate an ER 

stress response, we next tested cells lacking in each of the three 

canonical ER stress sensors, IRE1alpha, ATF6alpha and PERK. 

Both PA and Tg induced a significant EV release in cells lacking 

ATF6 alpha or PERK; however, PA and Tg-induced EV release 

was significantly suppressed in IRE1alpha KO cells. We tested 

the involvement of XBP1 signaling in this phenomenon and 

found a reduction in EV response in cells lacking XBP1. Next 

we measured ceramides in PA-induced EVs as they are implicated 

in EV biogenesis. PA-induced EVs were enriched in C16 

ceramide; this enrichment occurred in an IRE1alpha-dependent 

manner. Inhibition of de novo ceramide synthesis by myriocin 

ameliorated PA-induced EV release, and exogenous C16 ceramide 

resulted in EV release in IRE1alpha KO and WT cells. 

Lastly, PA-stimulated EVs were chemotactic to macrophages. 

Conclusions: PA induces C16 ceramide-enriched EV release 

in an IRE1alpha-dependent manner. PA-induced EVs stimulate 

macrophage chemotaxis and this may be a mechanism for the 

recruitment of macrophages to the liver under lipotoxic conditions. 

We hypothesize that interference with this macrophage 

recruitment response may be a therapeutic avenue in NASH. 

Disclosures: 

The following authors have nothing to disclose: Eiji Kakazu, Amy S. Mauer, 

Harmeet Malhi 

59 

Deficiency of intestinal mucin-2 protects mice from 

diet-induced fatty liver disease and obesity 

Phillipp Hartmann, Caroline T. Seebauer, Magdalena Mazagova, 

Angela Horvath, Lirui Wang, Cristina Llorente-Izquierdo, Katharina 

Brandl, Samuel B. Ho, David A. Brenner, Bernd Schnabl; University 

of California San Diego, La Jolla, CA 

Background: Non-alcoholic fatty liver disease (NAFLD) and 

obesity are characterized by altered gut microbiota, inflammation, 

and gut barrier dysfunction. The aim of this study was to 

investigate the role of mucin-2 (Muc2) as the major component 

of the intestinal mucus layer in the development of fatty liver 

disease and obesity. Methods and Results: We studied experimental 

fatty liver disease and obesity induced by high-fat diet 

(HFD) feeding for 16 weeks in wild-type (WT) littermate and 

mucin-2 knockout mice. Muc2 deficiency protected mice from 

HFD-induced obesity and fatty liver disease as evidenced by a 

significantly reduced body weight and lower hepatic triglyceride 

concentrations, respectively. Furthermore, Muc2 -/- mice displayed 

significantly decreased weights of subcutaneous and 

epididymal white adipose tissue and brown adipose tissue 

relative to WT mice on HFD. There was no difference in food 

intake and fecal triglyceride content between the HFD groups 

that would explain differences in phenotype. HFD-fed Muc2 -/- 

mice exhibited improved glucose tolerance and responsiveness 

to insulin, as evidenced by intraperitoneal glucose tolerance 

and insulin tolerance tests, as well as reduced inflammation 

in white adipose tissue than WT littermates. Furthermore, they 

showed an upregulated expression of key genes involved in 

lipolysis, such as adipose triglyceride lipase (ATGL) and hormone-sensitive 

lipase (HSL), and in fatty acid β-oxidation, e.g. 

palmitoyl acyl-CoA oxidase 1 (ACOX1), in white adipose tissue 

compared with WT mice after 16 weeks of HFD feeding. 

Interleukin-22 (IL-22) is not only crucial for maintaining intestinal 

homeostasis, but administration of IL-22 to obese mice has 

been shown to improve insulin sensitivity, decrease chronic 

inflammation and reduce body weight. Most interestingly, 

Muc2 -/- mice showed increased intestinal gene expression 

of IL-22 and IL-22 target genes Reg3b and Reg3g relative to 

WT mice fed HFD. Plasma levels of IL-22 were significantly 

higher in HFD-fed Muc2 -/- mice as compared with WT mice. 

After bypassing the mucus layer by intraperitoneal injections 

of bacteria-derived flagellin, differences in IL-22 plasma levels 

between HFD-fed WT and Muc2 -/- mice vanished, suggesting 

that the mucosal immune system produces increased amounts 

of IL-22 in the absence of a protective mucus barrier. Conclusion: 

An impaired gut barrier elicits a strong intestinal immune 

response in Muc2 deficient mice, which is characterized by the 

induction of IL-22. Increased systemic IL-22 mediates beneficial 

metabolic and anti-inflammatory effects, and protects against 

obesity and fatty liver disease. 

Disclosures: 

Angela Horvath - Grant/Research Support: Instutut Allergosan 

Samuel B. Ho - Grant/Research Support: Gilead, Genentech; Speaking and 

Teaching: Prime Education, Inc 

The following authors have nothing to disclose: Phillipp Hartmann, Caroline T. 

Seebauer, Magdalena Mazagova, Lirui Wang, Cristina Llorente-Izquierdo, Katharina 

Brandl, David A. Brenner, Bernd Schnabl


60 

FoxO3 increases miR-34a to cause palmitate-induced 

cholangiocytes lipoapoptosis 

Sathish Kumar Natarajan, Cody J. Wehrkamp, Ashley M. Mohr, 

Mary A. Smith, Sizhao Lu, Duygu Dee Harrison-Findik, Justin L. 

Mott; UNIVERSITY OF NEBRASKA MEDICAL CENTER, Omaha, 

NE 

Abstract: Non-alcoholic fatty liver disease (NAFLD) patients 

have elevated plasma saturated free fatty acid (FFA) levels. 

We had recently demonstrated the critical role of FoxO3 and 

its downstream target, PUMA in FFA-induced cholangiocyte 

lipoapoptosis and provided insights for biliary damage in a 

subset of NAFLD patients. Here, we explored whether miR-34a 

have a role in cholangiocyte lipoapoptosis. Methods: Apoptosis 

was assessed by TUNEL, characteristic nuclear morphology 

staining with DAPI, and caspase 3/7 activity assay. miR-34a 

levels were measured using quantitative real time-PCR. FoxO3, 

cleaved PARP, cleaved caspase 3 and miR-34a target expression 

was examined by Western blot. Results: Treatment of cholangiocytes 

with palmitate (PA) showed 7-20 fold increase in 

caspase3/7 activity in normal immortalized cholangiocytes, 

H69 and cell lines derived from cholangiocarcinoma, KMCH, 

Mz-ChA-1 and HuCCT cells. PA treatment also resulted in a dramatic 

increase in the levels of caspase cleaved products such 

as cleaved PARP and cleaved caspase 3 suggesting cholangiocyte 

lipoapoptosis. Interestingly, treatment with PA significantly 

increased levels of miR-34a in cholangiocytes and hepatocyte 

cell line, Huh7. PA-induced increase in miR-34a levels were 

abolished in cholangiocytes transduced with FoxO3 shRNA, 

implicating that miR-34a is a transcriptional target of FoxO3. 

Further, cholangiocytes transfected with anti-miR-34a were protected 

from PA-induced lipoapoptosis suggesting a major role 

for miR-34a. Further, direct and indirect targets of miR34a such 

as Sirt1, receptor tyrosine kinase (cMET), Kruppel like factor 4 

(KLF4), fibroblast growth factor receptor (FGFR1 and 4) were 

all decreased in PA-treated cholangiocytes. In addition, our 

data also shows that cholangiocyte apoptosis and miR-34a are 

dramatically increased in the liver from mice fed with high-fat 

high-sucrose for 3 months compared to control fed animals.In 

conclusion, PA induces the expression of proapoptotic miR-34a 

via FoxO3 and increased miR-34a levels results in targeting 

protein deacetylase Sirt1 and anti-apoptotic proteins like cMET 

and KLF4 resulting in cholangiocyte lipoapoptosis. 

Disclosures: 

The following authors have nothing to disclose: Sathish Kumar Natarajan, Cody 

J. Wehrkamp, Ashley M. Mohr, Mary A. Smith, Sizhao Lu, Duygu Dee Harrison-Findik, 

Justin L. Mott 

61 

Prior Blockade of TNF-α Signaling Promotes Liver 

Regeneration and Offers Strategies for Improving Outcomes 

after Liver Resection Surgery 

Fadi L. Jaber; GASTROENTEROLOGY AND LIVER DISEASES, 

ALBERT EINSTEIN COLLEGE OF MEDICINE, Bronx, NY 

The presumed beneficial role in liver regeneration (LR) of 

pro-inflammatory cytokines, such as TNF-α or IL6, contrasts 

with deleterious effects of these cytokines in other settings, 

e.g., after ischemia/reperfusion, inflammation, etc. Recently, 

cell transplantation-induced liver injury was shown to activate 

multiple chemokines/cytokines/receptors, which were essentially 

normalized after prior blockade of TNF-α by the drug, 

Etanercept (ETN), with superior transplanted cell engraftment 

and proliferation during liver repopulation in retrorsine (Ret)/ 

partial hepatectomy (PH) rat model. To resolve what role TNF-α 

may serve in LR, we hypothesized that administering ETN to 

block TNF-α before PH will allow determination of whether LR 

will be impaired or if it would still proceed. Groups of F344 

rats were established for 2/3 PH alone or ETN followed by 

2/3 PH along with untreated controls. Time-course analysis 

at 6, 32 or 72 h and 7 d after these procedures showed ETN 

was well tolerated and liver/body weight ratios were actually 

superior rather than inferior in ETN-pretreated rats with PH, 

p


experience a fibrotic phase dominated by inflammatory macrophages, 

followed by a phase of fibrosis remodelling characterised 

by restorative macrophages. Gpnmb- mice show a 

higher level of circulating hepatic enzymes in the phase of 

fibrosis remodelling, associated with higher proliferation of 

parenchymal cells and higher levels of inflammatory cytokines 

in the tissue. Moreover those mice show a lower number of 

macrophages and T cells in the phase of fibrosis resolution, 

determined by immunohistochemistry. Flow cytometry analysis 

unveiled a possible skew to a pro-inflammatory phenotype in 

the recovering liver. Key cytokines have been investigated as 

a possible cross-regulator of phagocytosis and tissue proliferation. 

The role of IL6 as a pro-proliferative agent in the liver 

is well known. No information is available on its role as a 

modulator of phagocytosis. Both in vivo and in vitro IL6 shows 

a pro-phagocytic effect possibly through the activation of the 

STAT3 pathway downstream the IL6 receptor. Collectively 

our data suggest that phagocytosis could be the gatekeeper 

between the necessary phase of initial inflammation and the 

following phase of tissue remodelling. 

Disclosures: 

The following authors have nothing to disclose: Lara Campana, Antonella Pellicoro, 

Rebecca Aucott, Stephen Greenhalgh, Katherine L. Hull, Stuart J. Forbes, 

John P. Iredale 

63 

Dynamic shift of microbiota and its relationship with 

hepatic gene expression during liver regeneration 

Hui-Xin Liu 1 , Clarissa S. Rocha 2 , Satya Dandekar 2 , Yu-Jui Yvonne 

Wan 1 ; 1 Medical Pathology and Laboratory Medicine, UC DAVIS, 

Sacramento, CA; 2 Medical Microbiology and Immunology, UC 

DAVIS, Davis, CA 

ABSTRACT: The pathways that regulate hepatocyte proliferation 

and liver regeneration have been extensively studied within 

the liver. However, the signaling contribution specifically from 

the gut microbiota involved in liver regeneration is poorly 

understood. We have temporally profiled the microbiota, bile 

acids (BA), and hepatic transcriptomes in regenerating livers 

obtained from mice 0, 1 hour, 1, 2, 3, 5, 7, and 9 days post 

2/3 partial hepatectomy (Phx) and established their interactive 

relationships. Our data showed that liver resection led 

to rapid changes in the gut microbiota that was reflected in 

increased abundance of Bacteroidetes including S24-7 and 

Rikenellaceae and decreased abundance of Firmicutes including 

Clostridiales, Lachnospiraceae, and Ruminococcaceae. 

These changes persisted throughout the course of liver regeneration. 

Short Time-series Expression Miner analysis of RNA-Sequencing 

data of 6,125 genes, which had greater than 2 folds 

changes at their mRNA levels compared day 0 controls, generated 

six unique patterns: peaked at day 1 (691), 2 (818), 

and 3 (275), dipped at day 2 (175), continuously increasing 

(340) or decreasing (558) during the course of regeneration. 

Correlation analyses revealed close associations between the 

expression of genes involved in BA homeostasis, metabolism, 

and immune function with the altered abundance of Ruminococcacea, 

Lachnospiraceae, and S24-7. Quantification of BA 

demonstrated a rapid increase of free and conjugated BAs 1 

hour after liver resection due to a sudden loss of liver mass. The 

hepatic conjugated BA overload was correlated with increased 

abundance of Bacteroidetes, which mediate BA conjugation 

by expressing 7α-dehydroxylase. The possibility of shifts in the 

microbiota profile as a consequence of changes in BA composition 

following liver resection remains to be determined. Conclusion: 

This is the first report to show that microbiota profile is 

altered during liver regeneration. Moreover, there appears to 

be a close relationship between microbiota composition and 

hepatic gene expression pattern. The data suggested a significant 

role of microbiota in contributing to the liver regeneration. 

Disclosures: 

The following authors have nothing to disclose: Hui-Xin Liu, Clarissa S. Rocha, 

Satya Dandekar, Yu-Jui Yvonne Wan 

64 

Annexin A6 is necessary for liver regeneration and glucose 

homeostasis in mice 

Carles Rentero Alfonso 1,2 , Anna Alvarez-Guaita 1 , Stephen E. 

Moss 3 , Thomas Grewal 4 , Carlos Enrich 1,2 ; 1 Biologia Cel.lular, 

Immunologia i Neurociences, Universitat de Barcelona, Barcelona, 

Spain; 2 Centre de Recerca Biomèdica CELLEX, Institut d’Investigacions 

Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; 

3 Department of Cell Biology, UCL Institute of Ophthalmology, London, 

United Kingdom; 4 Faculty of Pharmacy, University of Sydney, 

Sydney, NSW, Australia 

Annexin A6 (AnxA6) belongs to a conserved family of Ca 2+ - 

and phospholipid-binding proteins which interact with membranes 

upon increase of intracellular Ca 2+ concentration. We 

have previously shown the relevance of AnxA6 in the regulation 

of endocytic and exocytic pathways, intracellular cholesterol 

homeostasis, in the EGFR/PKCα/Ras/MAPK signaling 

pathway and in membrane remodeling. However, although 

AnxA6 is one of the major proteins in the liver (0.25% total 

protein), its function in the physiology of this organ remains 

unknown. After partial hepatectomy (PHx), an acute proliferative 

and metabolic stress, quiescent hepatocytes are triggered 

to progress through the cell cycle, showing a synchronous onset 

of DNA synthesis with a cellular response that involves cell 

activation and tissue remodeling. During this period of liver 

regeneration, the liver has to retain the major physiological 

tasks such as the synthesis and secretion of plasma proteins, 

lipid homeostasis and its metabolic function to ensure viability 

of the organism. The aim of the present study was to investigate 

the in vivo function of AnxA6 during liver regeneration using 

an AnxA6 knock-out (AnxA6 -/- ) mouse model. To this objective, 

2/3 PHx was performed in wild-type C57BL/6 control and 

AnxA6 -/- mice. Histological, electron microscopy, biochemical 

and metabolic studies after PHx were performed. Primary 

hepatocytes were also isolated and cultured to confirm the 

liver-specificity. After PHx, AnxA6 -/- mice exhibit a dramatic 

reduction of survival rate (87.5% at 72h post-PHx). However, 

the exogenous administration of glucose before and during 

PHx restored AnxA6 -/- survival rate after PHx, suggesting a link 

between AnxA6 and hepatic energetic metabolism. A comprehensive 

analysis of glucose metabolism experiments pointed to 

an impairment of liver gluconeogenesis in AnxA6 -/- mice, also 

observed in starving mice. A biochemical approach allowed us 

to elucidate a role for AnxA6 in the intracellular trafficking of 

SLC38A2, the major liver L-Alanine transporter, which is essential 

for gluconeogenic hepatic substrate incorporation during 

liver regeneration and mice starvation. We conclude that 

AnxA6 is a new regulator of hepatic gluconeogenesis essential 

for the trafficking of the alanine transporter SLC38A2 into 

the hepatocyte sinusoidal plasma membrane and subsequent 

alanine uptake, the major gluconeogenic substrate during both 

liver regeneration and fasting. 

Disclosures: 

The following authors have nothing to disclose: Carles Rentero Alfonso, Anna 

Alvarez-Guaita, Stephen E. Moss, Thomas Grewal, Carlos Enrich


65 

Akt-mediated FoxO1 inhibition is required for liver 

regeneration after partial hepatectomy in mice 

Montserrat Pauta 1,3 , Noemi Rotllan 4,5 , Ana Fernandez-Hernando 6 , 

Cedric Langhi 7 , Jordi Ribera 1,2 , Loreto Boix 8,2 , Jordi Bruix 8,2 , 

Wladimiro Jiménez 1,9 , Yajaira Suarez 4,5 , David A. Ford 7 , Angel 

Baldan 7 , Morris J. Birnbaum 10 , Manuel Morales-Ruiz 1,2 , Carlos 

Fernandez-Hernando 4,5 ; 1 Biochemistry and Molecular Genetics, 

Hospital Clinic, Barcelona, Spain; 2 IDIBAPS, CIBERehd, Barcelona, 

Spain; 3 IDIBAPS, Barcelona, Spain; 4 Vascular Biology and Therapeutics 

Program, Yale University School of Medicine, New Haven, 

CT; 5 Integrative Cell Signaling and Neurobiology of Metabolism 

Program, Section of Comparative Medicine and Department of 

Pathology, Yale University School of Medicine, New Haven, CT; 

6 Departments of Medicine and Cell Biology, Leon H. Charney 

Division of Cardiology, New York University School of Medicine, 

New York, NY; 7 Edward A. Doisy Department of Biochemistry & 

Molecular Biology, and Center for Cardiovascular Research, Saint 

Louis University, Saint Louis, MO; 8 Barcelona Clinic Liver Cancer 

(BCLC) Group, Liver Unit, Hospital Clinic of Barcelona, University 

of Barcelona, Barcelona, Spain; 9 Department of Physiological 

Sciences I, IDIBAPS, CIBERehd, University of Barcelona School of 

Medicine, Barcelona, Spain; 10 Institute for Diabetes, Obesity, and 

Metabolism, Perelman School of Medicine, University of Pennsylvania, 

Philadelphia, PA 

Introduction and aim: Understanding the hepatic regenerative 

process has clinical interest since the effectiveness of many 

treatments for chronic liver diseases is conditioned by an efficient 

liver regeneration. Experimental evidence points to the 

need of a temporal coordination between cytokines, growth 

factors and metabolic signaling pathways to enable successful 

liver regeneration. One intracellular mediator that acts as a 

signal integration node for these processes is the serine-threonine 

kinase Akt, being Akt1 and Akt2 the most abundant 

isoforms expressed in the liver. Therefore, the aim of this study 

was to demonstrate whether Akt, globally or through any of 

its isoforms, has a significant role in liver regeneration. Methods: 

Two-thirds partial hepatectomy (PH) was performed in 

eight-old-week male Akt1 -/- , Akt2 -/- , Akt1 loxP/loxP , Akt2 loxP/loxP , 

FoxO1 loxP/loxP and wild-type (WT) mice. In addition, we generated 

the Akt1 loxP/loxP ;Akt2 loxP/loxP ;FoxO1 loxP/loxP mice (TLKO) 

by crossing the Akt1 loxP/loxP ;Akt2 loxP/loxP mice (DLKO) with the 

FoxO1 loxP/loxP mice. The liver specific DLKO and TLKO deficient 

mice were generated by treating the floxed mice with adeno-associated 

virus encoding for the Cre-recombinase driven 

by the Alpha-Fetoprotein promoter. Mice from each group 

were sacrificed at various times post-hepatectomy (0 days, 2d, 

4d and 7d; n = 5-10) for evaluating biochemical and liver 

function parameters. Results: The hepatic deficiency of single 

Akt1 or Akt2 does not influence liver regeneration after PH. 

However, DLKO mice show impaired liver regeneration and 

a significant increase mortality (compared with WT, Akt1 -/- 

and Akt2 -/- mice; P10-fold; 

p50%) by 15min, 

and only approached basal levels by 96h post-PH. TMX-DTG 

mice (with reduced Hh signaling) accumulated 50% fewer 

Gli2+ cells, and 2-3 fold less Gli1 and Gli2 mRNA (p


approaches that inhibit Hh signaling down-regulate YAP, profibrogenic 

YAP targets and aSMA, showing that Hh regulates 

YAP and suggesting that YAP may be a downstream effector of 

pro-EMT Hh signaling. 

Disclosures: 

The following authors have nothing to disclose: Marzena Swiderska-Syn, Wing- 

Kin Syn, Guanhua Xie, Mark Jewell, Richard T. Premont, Gregory A. Michelotti, 

Anna Mae Diehl 

67 

Region-Based Projections of Deceased Liver Donors from 

2015-2025: An Analysis of Proposed UNOS Liver Allocation 

Redistricting 

Neehar D. Parikh 1 , Wesley J. Marrero Colon 2 , Yongcai Xu 2 , Anna 

S. Lok 1 , David W. Hutton 3 , Mariel S. Lavieri 2 ; 1 Division of Gastroenterology, 

University of Michigan, Ann Arbor, MI; 2 Industrial and 

Operations Engineering, University of Michigan, Ann Arbor, MI; 

3 School of Public Health, University of Michigan, Ann Arbor, MI 

Background: To help remedy the geographic disparity in liver 

transplant (LT) allocation the United Network of Organ Sharing 

(UNOS) has recently proposed regional redistricting plans. 

While current geographic inequity will improve, it is unclear to 

what extent demographic trends in the US will impact redistricting. 

We aimed to determine the impact of population demographic 

trends on liver redistricting proposals. Method: We 

performed a secondary analysis of the Organ Procurement 

and Transplantation Network database of all adult donors from 

2000 to 2012 and calculated the total number of donors available 

and transplanted donor livers stratified by age, race, and 

body mass index (BMI) group per year. We used National 

Health and Nutrition Examination Survey and Centers for Disease 

Control and Prevention historical data to stratify the US 

population by age, sex, race, and BMI. We then used US population 

age and race projections provided by the US Census 

Bureau and the Weldon Cooper Center for Public Service and 

made regional projections of available donors from 2015 to 

2025, incorporating the proposed 8- and 4-region redistricting 

plans proposed by UNOS. Given the uncertainty in LT demand, 

we used donors/100,000 population age 18-84 (D/100K) as 

a measure of equity. We calculated the mean and variation 

between regions, by calculating the percentage difference 

between the highest and lowest D/100K per allocation model. 

Results: The overall projected D/100K will decrease from 2.72 

to 2.65 over the next decade. The projected percentage variation 

in 2015 and 2025 in each model is shown in the Table. 

In the current 11-region allocation system, the 2015 range in 

D/100K is 2.45-2.89, and is projected to be 2.42-2.83 in 

2025. In the proposed 8-region allocation system the 2015 

range is 2.49-2.89, and is projected to be 2.42-2.83 in 2025. 

In the 4-region allocation system, the 2015 range is 2.49-2.79 

and is projected to be 2.42-2.71 in 2025. Conclusions: Of 

the current UNOS allocation schemes, the 4-region allocation 

model reduces the geographic variation to the greatest extent. 

This is projected to be maintained as the US population ages 

and undergoes demographic shifts over the next decade. However, 

this should be balanced by consequences of redistricting 

such as increased cold ischemia time and costs associated with 

organ transport. 

68 

Liver transplantation waiting list mortality in PSC 

patients is low as compared to non-PSC patients and 

consistent across laboratory MELD and MELD exception 

candidates: a nationwide study in the Netherlands 

Annemarie C. de Vries 1 , Madelon Tieleman 1 , Bart van Hoek 2 , 

Aad P. van den Berg 3 , Wojciech G. Polak 4 , Jan Ringers 5 , Robert J. 

Porte 6 , Cynthia Konijn 7 , Robert A. de Man 1 , Henk R. van Buuren 1 , 

Bettina E. Hansen 1 , Herold J. Metselaar 1 ; 1 Gastroenterology and 

Hepatology, Erasmus MC University Medical Center, Rotterdam, 

Netherlands; 2 Gastroenterology and Hepatology, Leiden University 

Medical Center, Leiden, Netherlands; 3 Gastroenterology and 

Hepatology, University Medical Center Groningen, Groningen, 

Netherlands; 4 Surgery, Erasmus MC University Medical Center, 

Rotterdam, Netherlands; 5 Surgery, Leiden University Medical 

Center, Leiden, Netherlands; 6 Surgery, University Medical Center 

Groningen, Groningen, Netherlands; 7 Dutch Transplantation Foundation, 

Leiden, Netherlands 

Background: PSC patients with end-stage disease form a heterogeneous 

group due to their varying complications. This 

hinders laboratory MELD score prioritization on the liver transplantation 

(LTx) waiting list, resulting in both lab MELD (LM) 

and MELD exception (ME) PSC candidates. Aim: To assess LTx 

waiting list mortality of LM and ME PSC candidates as compared 

to non-PSC patients. Methods: Patients aged ≥18 years 

who were listed for LTx in the Netherlands after introduction of 

the MELD score prioritization to December 31th 2013 were 

included. Data were recorded until November 2014. Exclusion 

criteria were reLTx, HU status or combined organ transplantation. 

A competing risk analysis was performed. Results: During 

the study period 852 candidates (M 579/ F 273; median age 

54.0 yrs) were listed for LTx. Median lab MELD score at listing 

was not significantly different between PSC patients (n=146) 

and non-PSC patients (n=706) (13.5 vs. 13.0; p=0.51). 

ME points were granted in resp. 22 PSC and 227 non-PSC 

patients (HR 0.34; p


Disclosures: 

Bart van Hoek - Advisory Committees or Review Panels: Janssen-Cilag, Bristol 

Meyers Squib, Gilead, Merck, Abbvie 

Robert A. de Man - Advisory Committees or Review Panels: Norgine; Grant/ 

Research Support: Biotest 

Henk R. van Buuren - Grant/Research Support: Intercept, Zambon Nederland BV 

The following authors have nothing to disclose: Annemarie C. de Vries, Madelon 

Tieleman, Aad P. van den Berg, Wojciech G. Polak, Jan Ringers, Robert J. Porte, 

Cynthia Konijn, Bettina E. Hansen, Herold J. Metselaar 

69 

Liver Transplant Outcomes and Survival Benefit for 

Obese Patients in the United States: Are We Disadvantaging 

the Morbidly Obese? 

Barry Schlansky 1 , Willscott E. Naugler 1 , Susan L. Orloff 2 , C. Kristian 

Enestvedt 2 ; 1 Gastroenterology/Hepatology, Oregon Health & 

Science University, Portland, OR; 2 Abdominal Organ Transplantation, 

Oregon Health & Science University, Portland, OR 

Purpose. Over 85% of U.S. centers adhere to national practice 

guidelines that consider morbid obesity to be a contraindication 

to liver transplantation (LT) based on inferior post-LT 

survival compared to the non-obese. In the present era, LT outcomes 

and survival benefit for obese patients have not been 

well studied. Methods. We evaluated the association of body 

mass index (BMI) with wait list and post-LT outcomes in patients 

wait listed for LT from 2005 to 2014, using the United Network 

for Organ Sharing database. We categorized BMI by the 

World Health Organization classification, with 18.5-29.9 kg/ 

m 2 defining normal weight, 30-34.9 obesity, 35-39.9 severe 

obesity, and ≥40 morbid obesity. We followed patients from 

LT to death or graft loss, from wait listing to death before LT 

or receipt of LT, and from wait listing to death before or after 

LT (intention-to-treat, ITT). We used Cox regression to evaluate 

post-LT and ITT mortality and LT survival benefit, and competing 

risk regression to evaluate wait list mortality versus receipt 

of LT. We also explored temporal trends of these outcomes 

in an expanded cohort from 2002. Results. 3.9% of 80,221 

waitlisted patients and 3.5% of 38,177 transplanted patients 

were morbidly obese. ITT survival was lower for morbidly 

obese compared to lower BMI patients (log-rank p


sis of complications during the donor hospitalization. Baseline 

data (PRE) 2/08-7/10 (29 months) were compared to donors 

receiving the pain protocol (POST) 3/13-4/15 (26 months). 

An external monitor validated the findings. Each center was IRB 

approved. RESULTS: 97 donors received SOC (PRE) and 75 

donors received the pain protocol (POST). Complications due 

to opioids were nausea (82%), hypoxia


73 

Incidence and Impact of Decompensating Events in Primary 

Biliary Cirrhosis - Results of an International Follow 

Up Study of 3030 Patients. 

Maren H. Harms 1 , Willem J. Lammers 1 , Harry L. Janssen 2 , Christophe 

Corpechot 3 , Pietro Invernizzi 4 , Marlyn J. Mayo 5 , Pier Maria 

Battezzati 6 , Annarosa Floreani 7 , Albert Pares 8 , Frederik Nevens 9 , 

Andrew Mason 10 , Kris V. Kowdley 11 , Cyriel Y. Ponsioen 12 , Tony 

Bruns 13 , George N. Dalekos 14 , Douglas Thorburn 15 , Gideon 

Hirschfield 16 , Nicholas F. LaRusso 17 , Ana Lleo 4 , Nora Cazzagon 

7 , Irene Franceschet 7 , Llorenc Caballeria 8 , Kalliopi Zachou 14 , 

Raoul Poupon 3 , Angela C. Cheung 2 , Palak J. Trivedi 16 , Marco 

Carbone 4 , Keith D. Lindor 18 , Henk R. van Buuren 1 , Bettina E. 

Hansen 1 ; 1 Gastroenterology and Hepatology, Erasmus University 

Medical Center, Rotterdam, Netherlands; 2 Liver Clinic, Toronto 

Western & General Hospital, University Health Network, Toronto, 

ON, Canada; 3 Centre de Référence des Maladies Inflammatoires 

des VoiesBiliaires, Hôpital Saint-Antoine, Paris, France; 4 Liver Unit 

and Center for Autoimmune Liver Diseases, Humanitas Clinical 

and Research Center, Rozzano, Italy; 5 Digestive and Liver diseases, 

UT Southwestern Medical Center, Dallas, TX; 6 Department 

of Health Sciences, Università degli Studi di Milano, Milan, Italy; 

7 Department of Surgery, Oncology and Gastroenterology, University 

of Padua, Padua, Italy; 8 Liver Unit, Hospital Clínic, CIBERehd, 

IDIBAPS, University of Barcelona, Barcelona, Spain; 9 Dept of 

Hepatology, University Hospitals Leuven, KULeuven, Leuven, Belgium; 

10 Divison of Gastroenterology and Hepatology, University of 

Alberta, Edmonton, AB, Canada; 11 Liver Care Network, Swedish 

Medical Center, Seattle, WA; 12 Dept of Gastroenterology and 

Hepatology, Academic Medical Center, Amsterdam, Netherlands; 

13 Department of Gastroenterology and Hepatology, University of 

Jena, Jena, Germany; 14 Institute of Internal Medicine and Hepatology, 

University of Thessaly, Larissa, Greece; 15 The Sheila Sherlock 

Liver Centre, The Royal Free Hospital, London, United Kingdom; 

16 NIHR Biomedical Research Unit and Centre for Liver Research, 

University of Birmingham, Birmingham, United Kingdom; 17 Dept 

Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN; 

18 Arizona State University, Phoenix, AZ 

Background Clinical events of decompensation are considered 

indicators of poor prognosis in primary biliary cirrhosis (PBC). 

Few studies have assessed the incidence of decompensating 

events and the related outcome of patients. Methods Long-term 

follow-up (FU) data of ursodeoxycholic acid (UDCA) treated 

patients were derived from 17 North-American and European 

centers. Decompensation was defined as a first event of ascites, 

variceal bleeding, or encephalopathy, whichever came 

first. Patients with events prior to baseline or in first year of FU 

were excluded. Risk factor analysis was performed using Cox 

proportional hazard models. Results Of 3030 UDCA-treated 

PBC patients, 92 patients (3.0%) were excluded. Median FU 

was 8.4 yrs (IQR 4.7-12.9). A decompensating event occurred 

in 275 patients: ascites:167 (61%), variceal bleeding:76 

(27%), encephalopathy:24 (9%), multiple:8 (3%). 1-, 3- and 

5- year transplantation-free survival with or without event was 

59% vs 99%, 35% vs 97% and 19% vs 94% respectively (time 

dependent HR40.6; 95%CI 29.6-55.7). Survival did not significantly 

differ between different types of events. Multivariable 

analysis showed that at time of first event, the following 

factors are predictive of survival: age at time of event (per 

10 yrs) (HR 1.39; 95%CI 1.09-1.63), calendar year of event 

(HR0.97; 95%CI 0.94-0.99), bilirubin>2x upper limit of normal 

(ULN) (HR2.98; 95%CI 1.99-4.45) and normal albumin 

(HR1.68; 95%CI 1.12-2.53). Survival of patients with normal 

albumin and bilirubin 

2xULN (p


LTx. However, prognostic models based on population-based 

cohorts are lacking. Various biomarkers of disease progression 

have been assessed for PSC, and the potential prognostic 

value of serum alkaline phosphatase (ALP) over time has been 

discussed. The aim of this study was to create a prognostic 

model for PSC consisting of disease characteristics and biochemical 

variables, and to validate this model in an external 

cohort. Methods 692 PSC patients were identified in a large, 

population-based PSC cohort from the Netherlands. Disease 

characteristics and biochemical variables during long term 

follow-up (FU) were retrieved from patient records. Clinical 

endpoints were: development of cholangiocarcinoma, LTx, 

or PSC-related death. Biochemical tests were transformed by 

log transformation, missing values were imputed by multiple 

imputation. All variables were assessed as potential predictors 

of survival by univariate analysis. To calculate the prognostic 

index (PI), a Cox proportional hazards model was developed 

and internally validated with bootstrap. The model was 

then validated in an external cohort of 259 PSC patients from 

UK. Results The median FU was 85 months (range 0-468 

months). All disease characteristics and biochemical variables 

were considered for the model. After variable selection by 

LASSO, multivariate Cox models were fitted, and parameters 

estimated from 20 imputation datasets were averaged. The 

following formula was created: PI=1.409*PSC type (0/1) 1 

+ 0.021*Age_PSC diagnosis - 2.420*log_AlbuminxULN 2 + 

2.073*abs(log_TrombocytesxULN-0.5) 2 + 0.469*log_AspartateAminotransferase(AST)xULN 

2 + 0.565*log_ALPxULN 2 + 

0.528*log_TotalBilirubinxULN 2 1: PSC type: Large duct=1; 

Small duct=0 2: xULN= times upper limit of normal A higher 

PI indicated a worse prognosis, corresponding to a shorter 

endpoint-free survival. The PI yielded a c-statistic of 0.715 in 

the development dataset, 0.705 in internal validation (adjusted 

for optimism with 1000 times bootstrap), and 0.683 in the 

external validation dataset. Conclusion A novel prognostic PSC 

model based on PSC type, age at PSC diagnosis, albumin, 

thrombocytes, AST, ALP and bilirubin which shows adequate 

performance in internal and external validation cohorts, allows 

to determine complication-free survival probability for PSC 

patients over time. 

Disclosures: 

Ulrich Beuers - Consulting: Intercept via University of Amsterdam, Novartis via 

University of Amsterdam; Grant/Research Support: Falk, Zambon; Speaking and 

Teaching: Falk Foundation, Gilead, Roche, Shire 

Cyriel Y. Ponsioen - Advisory Committees or Review Panels: Takeda; Consulting: 

AbbVIE; Grant/Research Support: AbbVIE, Schering Plough, Dr. Falk Pharma, 

Tramedico Netherlands, Takeda 

The following authors have nothing to disclose: Elisabeth M. de Vries, Junfeng 

Wang, Kate D. Williamson, Mariska M. Leeflang, Kirsten Boonstra, Roger W. 

Chapman, Ronald Geskus 

75 

Clinical Epidemiology of Primary Biliary Cirrhosis based 

on a Large US Laboratory Database: Incidence and 

Trends in Serum Alkaline Phosphatase 

W. Ray Kim 1 , Tracy J. Mayne 2 , Tonya Marmon 3 , David Shapiro 4 , 

Keith D. Lindor 5 ; 1 Division of Gastroenterology and Hepatology, 

Stanford University School of Medicine, Stanford, CA; 2 Outcomes 

Research, Intercept Pharmaceuticals, New York, NY; 3 Biostatistics, 

Intercept Pharmaceuticals, San diego, CA; 4 Office of the Chief 

Medical Officer, Intercept Pharmaceuticals, San Diego, CA; 5 College 

of Health Solutions, Arizona State University, Tempe, AZ 

Backround/Aims: Large scale epidemiological data on primary 

biliary cirrhosis (PBC) are scarce. Using a nationwide database 

covering approximately 30% of US adults, we estimate 

PBC incidence and describe trends in serum alkaline phosphatase 

(ALP) activities following the diagnosis. Methods: Electronic 

data from a large commercial laboratory were queried 

to extract all AMA and ALP results between Jan 2010 and Dec 

2013. Following AASLD guidelines, we defined probable PBC 

by positive anti-mitochondrial antibodies (AMA) and elevated 

serum ALP. AMA positivity was defined by a titer > 1:20 or M2 

antibody > 25.0 U, and elevated ALP (> upper limit of normal 

(ULN), 120 U/L) by the peak ALP before and up to 30-days 

after the first positive AMA. In subjects with PBC, serum ALP 

was followed for 24 months after the first positive AMA (baseline) 

and the highest ALP was identified per 6-month interval. 

Results: Out of over 576,000 persons receiving AMA testing, 

16,492 unique individuals (2.9%) tested positive. In 5,380 

(33%), serum ALP was elevated at baseline, while another 727 

AMA-positive patients developed elevated ALP subsequent to 

a positive AMA result. Thus, a total of 6,107 met the criteria 

for probable PBC, giving rise to a crude incidence rate of 

5.8 per 100,000 over 4 years. In the figure, of the 5,380 

patients, 47% had ALP > x2ULN, including 25% with ALP > 

3xULN. Following the diagnosis, the proportion of elevated 

ULN decreased, presumably as a result of therapy. By 3-6 

months, 61% of patients had ALP < 1.5xULN. Subsequent 

trends did not show further improvement in ALP – as of 24 

months post diagnosis, 69% had elevated ALP, including 35% 

with ALP > 1.5xULN. Conclusion: This largest epidemiological 

survey for PBC in US to date suggests that annually more than 

5000 Americans meet the diagnosis of probable PBC. Two 

years after the diagnosis, more than a third continue to have 

significantly elevated ALP. 

Disclosures: 



Tracy J. Mayne - Employment: Intercept Pharmaceuticals 

Tonya Marmon - Employment: Intercept Pharmaceuticals, Inc; Stock Shareholder: 

Intercept Pharmaceuticals, Inc 

David Shapiro - Employment: Inttercept Pharmaceuticals; Management Position: 

Intercept Pharmaceuticals; Stock Shareholder: Intercept Pharmaceuticals 

The following authors have nothing to disclose: Keith D. Lindor


76 

Gender and IBD phenotype are independent predictors 

of death or transplantation and of malignancy in Primary 

Sclerosing Cholangitis – a multicenter retrospective 

study of the International PSC Study Group (IPSCSG) 

Tobias J. Weismuller 1 , Bettina E. Hansen 2 , Palak J. Trivedi 3 , Mohamad 

Imam 6 , Henrike Lenzen 5 , Cyriel Y. Ponsioen 6 , Kristian Holm 7 , 

Ulrich Beuers 6 , Annika Bergquist 8 , Daniel Gotthardt 9 , Hanns-Ulrich 

Marschall 10 , Douglas Thorburn 11 , Rinse K. Weersma 12 , Johan 

Fevery 13 , Tobias Mueller 14 , Olivier Chazouillères 15 , Christoph 

Schramm 16 , Konstantinos Lazaridis 4 , Brian D. Juran 4 , Martti A. 

Färkkilä 19 , Stephen P. Pereira 18 , Sven Almer 17,8 , Cynthia Levy 20 , 

Andrew Mason 21 , Christopher L. Bowlus 22 , Annarosa Floreani 23 , 

Emina Halilbasic 24 , Michael Trauner 24 , Kidist K. Yimam 25 , Piotr 

Milkiewicz 26,27 , Dep K. Huynh 28 , Albert Pares 29 , Christine N. 

Manser 30 , George N. Dalekos 31 , Bertus Eksteen 32 , Gabi I. Kirchner 

38 , Christoph Sarrazin 37 , Vincent Zimmer 36 , Luca Fabris 35 , 

Pietro Invernizzi 34 , Felix Braun 33 , Marco Marzioni 39 , Christoph 

P. Berg 40 , Ansgar W. Lohse 16 , Gideon Hirschfield 3 , Christian 

P. Strassburg 1 , Michael P. Manns 5 , Keith D. Lindor 4 , Tom H. 

Karlsen 7 , Kirsten M. Boberg 7 ; 1 Department of Internal Medicine I, 

University of Bonn, Bonn, Germany; 2 Department of Gastroenterology 

and Hepatology, Erasmus University Medical Center, Rotterdam, 

Netherlands; 3 National Institute for Health Research (NIHR) 

Birmingham Liver Biomedical Research Unit (BRU) and Centre for 

Liver Research, University of Birmingham, Birmingham, United 

Kingdom; 4 Department of Gastroenterology and Hepatology, 

Mayo Clinic, Rochester, MN; 5 Department of Gastroenterology, 

Hepatology and Endocrinology, Hannover Medical School, Hannover, 

Germany; 6 Department of Gastroenterology & Hepatology, 

Academic Medical Center, Amsterdam, Netherlands; 7 Norwegian 

PSC Research Center and Section for Gastroenterology, Department 

of Transplantation Medicine, Division of Cancer Medicine, 

Surgery and Transplantation, Oslo University Hospital, Rikshospitalet, 

Oslo, Norway; 8 Center for Digestive Diseases, Division of 

Hepatology, Karolinska University Hospital and Karolinska Institutet, 

Stockholm, Sweden; 9 Dept. of Gastroenterology, Infectious 

Diseases and Intoxications, University Hospital Heidelberg, Heidelberg, 

Germany; 10 Department of Molecular and Clinical Medicine, 

Sahlgrenska Academy, University of Gothenburg, Gothenburg, 

Sweden; 11 The Sheila Sherlock Liver Centre and UCL Institute for 

Liver and Digestive Health, Royal Free Hospital, London, United 

Kingdom; 12 Department of Gastroenterology and Hepatology, University 

of Groningen and University Medical Center Groningen, 

Groningen, Netherlands; 13 Department of Hepatology, University 

Hospital Gasthuisberg, Leuven, Belgium; 14 Department of Internal 

Medicine, Hepatology and Gastroenterology, Charité Universitätsmedizin, 

Berlin, Germany; 15 Service d’Hépatologie, Hôpital Saint 

Antoine, Assistance Publique-Hôpitaux de Paris,Faculté de Médecine 

Pierre et Marie Curie, Paris, France; 16 1st Department of Medicine, 

University Medical Center Hamburg Eppendorf, Hamburg, 

Germany; 17 Gastroenterology & Hepatology, Linköping University, 

Linköping, Sweden; 18 Institute for Liver and Digestive Health, UCL, 

London, United Kingdom; 19 Division of Gastroenterology, Department 

of Medicine, Helsinki University Central Hospital, Helsinki, 

Finland; 20 University of Miami, Miami, FL; 21 Division of Gastroenterology 

and Hepatology, University of Alberta, Edmonton, AB, 

Canada; 22 Division of Gastroenterology and Hepatology, University 

of California Davis, Davis, CA; 23 Department of Surgery, 

Oncology and Gastroenterology, University of Padova, Padova, 

Italy; 24 Division of Gastroenterology and Hepatology, Department 

of Internal Medicine III, Medical University of Vienna, Vienna, 

Austria; 25 Department of Hepatology and Liver Transplantation, 

California Pacific Medical Center, San Francisco, CA; 26 Department 

of Clinical and Molecular Biochemistry, Pomeranian Medical 

University, Szczecin, Poland; 27 Liver and Internal Medicine Unit, 

Department of General, Transplant and Liver Surgery, Medical 

University of Warsaw, Warsaw, Poland; 28 Department of Gastroenterology 

and Hepatology, Royal Adelaide Hospital, Adelaide, 

SA, Australia; 29 Liver Unit, Hospital Clinic, IDIBAPS, CIBERehd, 

University of Barcelona, Barcelona, Spain; 30 Division for Gastroenterology 

and Hepatology, University Hospital Zurich (USZ), Zurich, 

Switzerland; 31 Department of Medicine and Research Laboratory 

of Internal Medicine, School of Medicine, University of Thessaly, 

Larissa, Greece; 32 University of Calgary, Snyder Institute for 

Chronic Diseases, Alberta, AB, Canada; 33 Campus Kiel, UKSH, 

Kiel, Germany; 34 Humanitas Clinical and Research Center, Center 

for Autoimmune Liver Diseases, Rozzano (MI), Italy; 35 Department 

of Molecular Medicine, University of Padua School of Medicine, 

Padua, Italy; 36 Department of Medicine II, Saarland University 

Medical Center, Homburg, Germany; 37 Department of Internal 

Medicine 1, Johann Wolfgang Goethe-University Hospital, Frankfurt, 

Germany; 38 Department of Internal Medicine 1, University 

Hospital of Regensburg, Regensburg, Germany; 39 Department of 

Gastroenterology, Università Politecnica delle Marche, Ancona, 

Italy; 40 Department of Gastroenterology, Hepatology, and Infectiology, 

Medical Clinic, University of Tübingen, Tübingen, Germany 

Primary sclerosing cholangitis (PSC) is a male predominant 

disease bearing a strong association with inflammatory bowel 

disease (IBD). The aims of this study were to address impact 

of gender and IBD phenotype on liver-related outcomes across 

a large, internationally representative multi-centre registry. 

All patients diagnosed with PSC between 1/1/1980 and 

12/31/2010 in 37 institutions from 17 countries were included 

in the study. Data on clinical presentation, survival, liver transplantation 

(LT), IBD, colorectal neoplasia and hepatobiliary 

malignancy (HBM) were collected. Cox regression analyses, 

stratified by region and adjusted for age, year of diagnosis, 

diagnose (PSC/small duct PSC/ with AIH overlap) with IBD 

as a time dependent covariate were used to study the effect of 

gender on the risk of HBM, colorectal neoplasia and the risk 

of LT and death. 7119 patients were included (65.5% male, 

mean age at diagnosis 37.4 years in males and 40.4 years 

in females (p


Disclosures: 

Palak J. Trivedi - Grant/Research Support: Wellcome Trust 







Hanns-Ulrich Marschall - Consulting: Albireo 

Olivier Chazouillères - Consulting: APTALIS, MAYOLY-SPINDLER 

Andrew Mason - Advisory Committees or Review Panels: AbbVie, Novartis, 

Intercept; Grant/Research Support: Abbvie, Gilead, Astellas 

Christopher L. Bowlus - Advisory Committees or Review Panels: Gilead Sciences, 

Inc; Grant/Research Support: Gilead Sciences, Inc, Intercept Pharmaceuticals, 

Bristol Meyers Squibb, Takeda, Lumena, Merck; Speaking and Teaching: Gilead 

Sciences, Inc 

Michael Trauner - Advisory Committees or Review Panels: MSD, Janssen, Gilead, 

Abbvie; Consulting: Phenex; Grant/Research Support: Intercept, Falk Pharma, 

Albireo; Patent Held/Filed: Med Uni Graz (norUDCA); Speaking and Teaching: 

Falk Foundation, Roche, Gilead 

Christoph Sarrazin - Advisory Committees or Review Panels: Bristol-Myers 

Squibb, Janssen, Merck/MSD, Gilead, Roche, Abbvie, Janssen, Merck/MSD; 

Consulting: Merck/MSD, Merck/MSD; Grant/Research Support: Abbott, Roche, 

Merck/MSD, Gilead, Janssen, Abbott, Roche, Merck/MSD, Qiagen; Speaking 

and Teaching: Gilead, Novartis, Abbott, Roche, Merck/MSD, Janssen, Siemens, 

Falk, Abbvie, Bristol-Myers Squibb, Achillion, Abbott, Roche, Merck/MSD, Janssen 

Gideon Hirschfield - Advisory Committees or Review Panels: Intercept Pharma; 

Consulting: Dignity Sciences, GSK, NGM Bio, Lumena, J & J; Grant/Research 

Support: BioTie; Speaking and Teaching: Falk Pharma 

Christian P. Strassburg - Advisory Committees or Review Panels: Novartis, Roche; 

Speaking and Teaching: Novartis, Merz, MSD, Falk Pharma, BMS, Abbvie 

Michael P. Manns - Consulting: Roche, BMS, Gilead, Boehringer Ingelheim, 

Novartis, Idenix, Achillion, GSK, Merck/MSD, Janssen, Medgenics; Grant/ 

Research Support: Merck/MSD, Roche, Gilead, Novartis, Boehringer Ingelheim, 

BMS; Speaking and Teaching: Merck/MSD, Roche, BMS, Gilead, Janssen, GSK, 

Novartis 

The following authors have nothing to disclose: Tobias J. Weismuller, Bettina 

E. Hansen, Mohamad Imam, Henrike Lenzen, Kristian Holm, Annika Bergquist, 

Daniel Gotthardt, Douglas Thorburn, Rinse K. Weersma, Johan Fevery, 

Tobias Mueller, Christoph Schramm, Konstantinos Lazaridis, Brian D. Juran, 

Martti A. Färkkilä, Stephen P. Pereira, Sven Almer, Cynthia Levy, Annarosa 

Floreani, Emina Halilbasic, Kidist K. Yimam, Piotr Milkiewicz, Dep K. Huynh, 

Albert Pares, Christine N. Manser, George N. Dalekos, Bertus Eksteen, Gabi 

I. Kirchner, Vincent Zimmer, Luca Fabris, Pietro Invernizzi, Felix Braun, Marco 

Marzioni, Christoph P. Berg, Ansgar W. Lohse, Keith D. Lindor, Tom H. Karlsen, 

Kirsten M. Boberg 

77 

Ig Repertoire of Autoantigen-Specific B cells in Primary 

Biliary Cirrhosis 

Weici Zhang 1 , Patrick S. Leung 1 , Ying Sun 1,2 , Thomas P. Kenny 1 , 

Guo-Xiang Yang 1 , Xiao-Song He 1 , Christopher L. Bowlus 3 , Ross 

L. Coppel 4 , Ignacio Sanz 6 , Aftab A. Ansari 5 , M. Eric Gershwin 1 ; 

1 Internal Medicine, University of California, Davis, CA; 2 Hepatology, 

Center for Non-Infectious Liver Diseases, Beijing 302 Hospital, 

Beijing, China; 3 Division of Gastroenterology and Hepatology, 

University of California at Davis, Sacramento, CA; 4 Department 

of Microbiology, Monash University, Melbourne, VIC, Australia; 

5 Department of Pathology, Emory University School of Medicine, 

Atlanta, GA; 6 Department of Medicine, Division of Rheumatology 

and Lowance Center for Human Immunology, Emory University, 

Atlanta, GA 

Background: Autoantibodies to the E2 subunit of pyruvate 

dehydrogenase (PDC-E2) are the serological hallmark of PBC. 

Interestingly, a subpopulation of anti-PDC-E2 autoantibodies 

are cross-reactive to chemical xenobiotics including 2-octynoic 

acid (2OA) and 6, 8-bis (acetylthio) octanoic acid (SAc), both 

putative etiological agents of PBC. Recent studies suggest that 

anti-PDC-E2 is derived from de novo activated autoantigen-specific 

B cells, or from B cells previously primed by xenobiotics 

when self-tolerance was originally compromised. Examination 

of antigen specific plasmablasts from PBC demonstrate an 

extraordinary frequency of circulating plasmablasts specific 

for PDC-E2 without significant detection of reactivity to xenobiotic 

antigens; this is in striking contrast to comparable sera 

analysis. We propose that circulating PDC-E2 plasmablasts 

result from positive selection to xenobiotics, i.e. the “original 

sin” was the creation of a neoantigen between a chemical 

xenobiotic and a self peptide. Our aim herein was to identify 

Ig sequences from plasmablasts directed to PDC-E2. Methods: 

1,680 single plasmablast cells (CD3 – CD19 + CD20 lo/– CD27 hi C- 

D38 hi ) were obtained from PBC with high PDC-E2-specific 

plasmablasts (>30%). Thereafter, Ig heavy chain (IgH) and Igk 

or Igl light chain (IgL) gene transcripts were amplified using a 

IgG, IgA and IgM H and L chain gene specific primer cocktail 

and sequenced. A total of 420 functional paired IgH and 

Igk or Igl gene sequences were analyzed. CDR3 length and 

frequency of positively and negatively charged residues in 

various V gene libraries were analyzed. Differential analysis 

was performed comparing IgM versus IgG or IgA gene libraries. 

Results: Our data revealed a decreased diversity of IgH 

and IgL usage with preferential usage of IGHV3-23, IGHV3- 

30, IGHV3-7, IGHV4-39, IGHV3-15, IGHV1-18, IGHV3-74 

and IGKV3-20, IGKV2-28. Moreover, the usage of IGHV3- 

23 and IGHV3-30 increased by two-fold in PBC compared to 

controls. Thirty-two clonal V region sequences in the repertoire 

were identified and shared among PBC patients. Interestingly, 

the unique rearrangements accounted for 0.84% of the total 

sequenced IgH rearrangements and the majority of sequences 

are highly mutated with an average of 17 replacement mutations 

per sequence. Conclusion: We suggest that the original 

breach of tolerance was secondary to molecular mimicry in 

a genetically susceptible host to a neoantigen that includes a 

chemical xenobiotic of PDC-E2; subsequently, through determinant 

spreading and autoantibody maturation, the predominant 

hallmark of PBC arises, autoantibodies directed at the inner 

lipoyl domain of PDC-E2. 

Disclosures: 




Sciences, Inc 

Ignacio Sanz - Advisory Committees or Review Panels: Pfizer; Consulting: Genentech, 

Sanofi; Grant/Research Support: MedImmune 

The following authors have nothing to disclose: Weici Zhang, Patrick S. Leung, 

Ying Sun, Thomas P. Kenny, Guo-Xiang Yang, Xiao-Song He, Ross L. Coppel, 

Aftab A. Ansari, M. Eric Gershwin


78 

Intrahepatic Cholestasis of Pregnancy (ICP) in U.S. Latinas 

and Chileans: Ancestry Analysis, Admixture Mapping, 

and Biochemical Features 

Laura Bull 1,2 , Donglei Hu 2,3 , Sohela Shah 1,2 , Luisa Temple 1,2 , 

Karla Silva 4,5 , Scott Huntsman 2,3 , Jennifer Melgar 1,2 , Mary 

Geiser 1,2 , Ukina Sanford 1,2 , Juan Ortiz 5,6 , Richard Lee 7 , Juan P. 

Kusanovic 4,5 , Elad Ziv 3,2 , Juan Vargas 8,9 ; 1 Medicine, San Francisco 

General Hospital, UCSF, San Francisco, CA; 2 Institute for 

Human Genetics, UCSF, San Francisco, CA; 3 Medicine, UCSF, 

San Francisco, CA; 4 Center for Research and Innovation in Maternal-Fetal 

Medicine, Hospital Dr. Sótero del Río, Puente Alto, Chile; 

5 Obstetrics and Gynecology, Hospital Dr. Sótero del Río, Puente 

Alto, Chile; 6 Obstetrics and Gynecology, Clínica Santa María, 

Santiago, Chile; 7 Obstetrics and Gynecology, Los Angeles County 

+ University of Southern California Medical Center, Los Angeles, 

CA; 8 Obstetrics, Gynecology and Reproductive Sciences, UCSF, 

San Francisco, CA; 9 Radiology, UCSF, San Francisco, CA 

Purpose: In the Americas, women with Indigenous American 

ancestry are at increased risk of intrahepatic cholestasis of 

pregnancy (ICP). We hypothesized that ancestry-related 

genetic factors contribute to this increased risk, and that the 

statistically powerful genetic admixture mapping approach 

could allow mapping of an ICP susceptibility locus. We considered 

that diagnosis, features, and management of ICP might 

differ between U.S. and Chilean clinical sites. Methods: We 

collected patient data and performed biochemical assays and 

SNP genotyping on samples from U.S. Latinas and Chilean 

women, with and without ICP. The study sample included 198 

women with ICP (90 from the U.S. and 108 from Chile) and 

174 pregnant control women (69 from the U.S. and 105 from 

Chile). We compared overall genetic ancestry between cases 

and controls, and performed genome-wide admixture mapping, 

taking country of enrollment into account, to screen for 

ICP susceptibility loci. We characterized features of ICP at the 

U.S. and Chilean clinical sites. Results: Cases had a greater 

proportion of Indigenous American ancestry than did controls 

(p=0.034); when U.S. and Chilean study samples were analyzed 

separately, this difference was apparent in the U.S., but 

not the Chilean, study sample, potentially due to differences in 

population history. Admixture mapping allowed identification 

of one locus for which Native American ancestry was associated 

with increased risk of ICP at a genome-wide level of 

significance (P = 3.1 x 10 -5 , P corrected 

= 0.035). This locus, on 

chromosome 2, has an odds ratio of 4.48 (95% CI: 2.21-9.06) 

for 2 versus zero Indigenous American chromosomes; the 10 

Mb 95% confidence interval does not contain any previously 

identified hereditary ‘cholestasis genes.’ ICP appears biochemically 

more severe in the U.S., than Chilean, study sample. 

Differences in management include 1) ursodeoxycholate was 

prescribed to a greater proportion of U.S. than Chilean patients 

(48% vs 1%); 2) most U.S. cases were delivered by 37 weeks 

gestation (or at time of diagnosis if later), while induction of 

labor is not usually performed until 40 weeks at the Chilean 

site; and 3) cesarean birth is more common in Chilean cases 

than controls, while rates are similar between U.S. cases and 

controls. Conclusions: Our results indicate that genetic factors 

contribute to risk of developing ICP in the Americas, and support 

the utility of clinical and genetic studies of ethnically mixed 

populations for increasing our understanding of ICP. Greater 

characterization of diagnosis, management and outcomes of 

ICP in different countries may improve our understanding of 

ICP, and its potential subtypes. 

Disclosures: 

Sohela Shah - Employment: Qiagen Bioinformatics 

The following authors have nothing to disclose: Laura Bull, Donglei Hu, Luisa Temple, 

Karla Silva, Scott Huntsman, Jennifer Melgar, Mary Geiser, Ukina Sanford, 

Juan Ortiz, Richard Lee, Juan P. Kusanovic, Elad Ziv, Juan Vargas 

79 

The effects of apoptosis inhibition on pulmonary alveolar 

type 2 epithelial cell alterations in experimental 

hepatopulmonary syndrome after common bile duct 

ligation 

Wenli Yang, Bingqian Hu, Michael B. Fallon, Junlan Zhang; The 

university of Texas Health Science Center, Houston, TX 

Introduction: Abnormal oxygenation due to lung ventilation-perfusion 

mismatch drives the development of hepatopulmonary 

syndrome (HPS). In addition to alterations in microvascular 

perfusion, dysfunction in type II alveolar epithelial cells (AT2), 

which play a critical role in maintaining alveolar ventilation 

through surfactant protein (SPs) production, develops in experimental 

HPS. Specifically, AT2 cell apoptosis and decreased 

SP production associated with increased circulating bile acid 

levels and bile acid nuclear receptor (FXRα) activation, reduce 

alveolar airspace and impair ventilation. However, the relative 

effects of FXRα activation on AT2 cell apoptosis and SP 

expression and whether apoptosis inhibition influences AT2 

cells and the development of HPS are unknown. Aim: To evaluate 

the effects of apoptosis inhibition on FXRα activation in 

AT2 cells and in experimental HPS. Methods: A murine AT2 

cell line (MLE-12) was treated with a FXRα agonist (GW4064, 

1-10mM) in the presence or absence of a pan caspase inhibitor 

(Z-VAD-FMK). SD rats underwent common bile duct ligation 

(CBDL) or sham operation and were treated with Z-VAD-FMK 

(1.0mg/kg/day, IP, for 3 weeks). Lung and cell apoptosis was 

determined by TUNEL staining or cleaved caspase-3 levels. 

Co-staining of TUNEL and SP-C was performed to assess lung 

AT2 cell apoptosis. SP levels were assessed by westerns or qRT- 

PCR. Lung morphology (alveolar mean chord length, Lm) and 

gas exchange (PO 2 

) were measured to evaluate HPS. Results: 

GW4064 treatment in MLE-12 cells caused a dose dependent 

increase in cleaved caspase-3 protein expression and reduction 

of SP-C and SP-B mRNAs. Z-VAD-FMK pretreatment attenuated 

GW4064 induced cleaved caspase-3 protein increases 

(76.0±6.1% reduction, p


80 

Statins activate the canonical hedgehog-signaling and 

aggravate non-cirrhotic portal hypertension, but inhibit 

the non-canonical hedgehog signaling and cirrhotic portal 

hypertension. 

Frank E. Uschner 1 , Ganesh Ranabhat 1 , Michaela Granzow 1 , Steve 

S. Choi 2 , Sabine Klein 1 , Robert Schierwagen 1 , Esther Raskopf 1 , 

Sebastian Gautsch 1 , Peter F. Van der Ven 3 , Christian P. Strassburg 

1 , Dieter O. Fürst 3 , Kanishka Hiththetiya 4 , Tilman Sauerbruch 1 , 

Anna Mae Diehl 2 , Jonel Trebicka 1 ; 1 Medical Clinic I, University of 

Bonn, Bonn, Germany; 2 Department of Medicine, Duke University, 

Durham, NC; 3 Department of Molecular Cell Biology, University of 

Bonn, Bonn, Germany; 4 Institute of Pathology, University of Bonn, 

Bonn, Germany 

Background: Portal hypertension is a source of complications in 

patients with liver cirrhosis, as well as in patients with portal vein 

obstruction. In both conditions, increased outflow resistance 

and hyperperfusion of splanchnic vessels trigger angiogenesis. 

Extrahepatic angiogenesis exacerbates hyperperfusion of 

the splanchnic vessels and aggravates portal hypertension. In 

cirrhosis, intrahepatic angiogenesis is involved in progression 

of fibrosis since activated hepatic stellate cells (HSC) produce 

collagen and serve as pericytes. HSC activation and activity 

are promoted by Hedgehog (Hh) pathway, which further regulates 

vascular integrity and angiogenesis in different tissues. 

Statins not only blunt liver fibrosis and decreases portal pressure, 

but also interacts with the Hh. This study investigated the 

differences of angiogenesis in cirrhotic and non-cirrhotic portal 

hypertension with special emphasis on the canonical (Shh/Gli) 

and non-canonical (Shh/RhoA) pathway. Methods: Cirrhotic 

(BCL; CCl 4 

) and non-cirrhotic (partial portal vein ligation/ 

PPVL) rats received either atorvastatin (15mg/kg, 7d) or control 

chow. Invasive hemodynamics were assessed by coloured 

microspheres. Implantation of matrigel (s.c. and i.p.) assessed 

angiogenesis in vivo. In vitro angiogenesis was analyzed in 

primary rat HSC and LX-2 cells, after atorvastatin incubation 

and transfection with RhoA plasmids. Expression was analyzed 

using RT-PCR and Western blot. Results: Atorvastatin decreased 

portal pressure, shunt flow and angiogenesis in cirrhotic rats, 

whereas atorvastatin increased these parameters in PPVL rats. 

Hh was upregulated in experimental and human liver cirrhosis, 

but was blunted by atorvastatin. The transfection of LX-2 cells 

with a RhoA plasmid increased Hh protein levels and inhibition 

of the Hh-pathway reduced profibrotic markers, which clearly 

indicate a non-canonical activation of Hh-pathway in HSC. 

Moreover, atorvastatin blocked this non-canonical Hh-pathway 

RhoA-dependently in activated HSCs. Interestingly, hepatic and 

extrahepatic Hh-pathway was enhanced in PPVL rats, which 

resulted in increased angiogenesis. Discussion: In summary, 

statins caused contrary effects in cirrhotic and non-cirrhotic 

portal hypertension. Atorvastatin inhibited the non-canonical 

Hh-pathway and angiogenesis in cirrhosis. In portal vein 

obstruction, statins enhanced the canonical Hh-pathway and 

aggravated PHT and angiogenesis. In conclusion, the broad 

use of statins with its beneficial effects in liver cirrhosis still 

requires caution in therapy of portal hypertension without cirrhosis. 

Disclosures: 



The following authors have nothing to disclose: Frank E. Uschner, Ganesh 

Ranabhat, Michaela Granzow, Steve S. Choi, Sabine Klein, Robert Schierwagen, 

Esther Raskopf, Sebastian Gautsch, Peter F. Van der Ven, Dieter O. Fürst, 

Kanishka Hiththetiya, Tilman Sauerbruch, Anna Mae Diehl, Jonel Trebicka 

81 

Hepatocyte Tissue Factor Contributes to the Hypercoagulable 

State in a Mouse Model of Chronic Liver Injury 

Pierre-Emmanuel Rautou 1,2 , Kohei Tatsumi 3 , Silvio Antoniak 3 , 

Albert P. Owens III 3 , Erica Sparkenbaugh 3 , Anna K. Kopec 4 , Rafal 

Pawlinski 3 , James P. Luyendyk 4 , Nigel Mackman 3 ; 1 Hepatology, 

Hôpital Beaujon - APHP, Clichy, France; 2 Inserm U970; Paris cardiovascular 

Research center @ HEGP, Paris, France; 3 Department 

of Medicine, Division of Hematology and Oncology, McAllister 

Heart Institute, University of North Carolina at Chapel Hill, Chapel 

Hill, NC; 4 Department of Pathobiology and Diagnostic Investigation, 

Michigan State University, East Lansing, MI 

Background & Aims: Patients with chronic liver disease and cirrhosis 

have a dysregulated coagulation system and are prone 

to thrombosis. The basis for this so-called procoagulant imbalance 

is not completely understood. Tissue factor (TF) is the 

primary initiator of coagulation in vivo. Patients with cirrhosis 

have increased TF activity in white blood cells and circulating 

microparticles. The aim of our study was to determine the role 

of TF and the contribution of different cellular sources of TF to 

the hypercoagulable state in a mouse model of chronic liver 

injury. Methods: We measured levels of TF activity in the liver, 

white blood cells and microparticles, and a marker of activation 

of coagulation [thrombin-antithrombin complexes (TATc)] 

in the plasma of mice subjected to bile duct ligation for 12 

days. We used wild-type mice, mice with a global TF deficiency 

(low TF mice), and mice deficient for TF in either myeloid 

cells (TF flox/flox ,LysM Cre mice) or in hepatocytes (TF flox/flox /Albumin 

Cre ). Data are presented as median (interquartile range). 

Results: Wild-type mice with liver injury had increased levels 

of TATc [9.7 (7.6-10.5) vs. 6.0 (5.7-7.0) ng/mL; p


82 

The association of non-selective beta-blocker use and 

acute kidney injury in patients with decompensated 

cirrhosis admitted into hospital - A Study from the North 

American Consortium for the Study of End Stage Liver 

Diseae (NACSELD). 

Florence Wong 1 , Jacqueline G. O’Leary 2 , K. Rajender Reddy 3 , 

Guadalupe Garcia-Tsao 4 , Scott W. Biggins 5 , Michael B. Fallon 6 , 

Puneeta Tandon 7 , Ram M. Subramanian 8 , Benedict Maliakkal 9 , 

Paul J. Thuluvath 10 , Hugo E. Vargas 11 , Patrick S. Kamath 12 , Leroy 

Thacker 13 , Jasmohan S. Bajaj 14,15 ; 1 Medicine, University of 

Toronto, Toronto, ON, Canada; 2 Medicine, Hepatology, Baylor 

University Medical Center, Dallas, TX; 3 Gastroenterology & Hepatology, 

University of Pennsylvania, Philadelphia, PA; 4 Digestive 

Diseases, Yale University School of Medicine, New Haven, CT; 

5 Gastroenterology, University of Colorado, Denver, CO; 6 Gastroenterology, 

Hepatology & Nutrition, University of Texas Health Science 

Center, Houston, TX; 7 Medicine, Gastroenterology, University 

of Alberta, Edmonton, AB, Canada; 8 Gastroenterology, Emory 

University Medical Centre, Atlanta, GA; 9 Gastroenterology, University 

of Rochester, Rochester, NY; 10 Institute of Digestive Health 

and Liver Disease, Mercy Hospital, Baltimore, MD; 11 Gastroenterology 

& Hepatology, Mayo Clinic, Scottsdale, AZ; 12 Internal 

Medicine, Gastroenterology & Hepatology, Mayo Clinic, Rochester, 

MN; 13 Biostatistics, Virginia Commonwealth University, Richmond, 

VA; 14 Gastroenterology, Hepatology & Nutrition, Virginia 

Commonwealth University, Richmond, VA; 15 Gastroenterology, 

McQuire VA Medical Center, Richmond, VA 

Non-selective beta-blocker (NSBB) use may be detrimental 

in patients with cirrhosis & refractory ascites, although this 

remains controversial. Theoretically, NSBB use in decompensated 

cirrhosis (DC) may decrease renal perfusion and predispose 

patients to renal impairment. Aim: To determine the 

impact of NSBB use on the risk i) for acute kidney injury (AKI) 

in hospitalized patients with DC, and ii) of infection to potentially 

accentuate this risk. Methods: NACSELD is a 16-center 

consortium that prospectively enrols non-elective cirrhotic inpatients. 

The presence of AKI (defined by Angeli et al, Gut 2015) 

at admission or its development during admission was evaluated 

and compared between those taking NSBB (NSBB+) 

vs those not (NSBB-) at admission. Further comparison was 

made between NSBB+ patients ± an infection during admission. 

Results: 981 cirrhotics (57±10 years, men: 64.6%, 

MELD: 19.5±7.3) with mostly alcoholic (43%) and hepatitis 

C (22.4%) cirrhosis were enrolled. Significantly more NSBB+ 

patients had a history of variceal bleeding and diabetes 

(p


of AKI. Peak AKIN stage reached was stage III:II:I in 80(66%): 

31(25%):11(9%) patients, respectively. Presence of any E-OF 

was strongly associated with both development of new AKI 

(p=0.004, OR 6.7, 95% CI 1.86-24.1) and progression of 

AKI (p=0.04, OR 2.28, 95%CI 1.01-5.1). This risk further 

increased with increase in the number of E-OFs (p=0.001, OR 

1.41, 95%CI 1.2-1.7). Further, AKI at admission alone was not 

associated with mortality, but predicted mortality together with 

any E-OF (p150000) and liver stiffness 

(LS) ( 60% and an IQR/ 

Median < 30% were considered. SS was US guided. Results: 

99 patients were included: 63% women; age 59 (± 6.6) years; 

BMI 28 (± 4.8); MELD 9.7 (± 3.4); 80% Child-Pugh A. LS was 

invalid in 2 patients and SS in 19. Platelets was 136.000 (± 

54.000), LS was 24 kPa (± 12) and SS was 53 kPa (± 18). 

Esophageal varices were detected in 54% of patients, 40% 

small and 14% large size. Gastric varices were present in 5% 

of patients, and GEV in 55%. Platelets (120.000 ± 45.000 

vs 155.000 ± 57.000), LS (28.4 ± 13.1 vs 18.4 ± 7.6 kPa) 

and SS (59.5 ± 17.0 vs 43.3 ± 17.9 kPA) were significantly 

different among patients with or without GEV (p 

6 months, n=83); and G4, past HCV infection (positive anti- 

HCV and negative HCV RNA PCR, n=15). The serum samples 

were tested using our HCV-Ags EIA before and after denaturation. 

RESULTS. All 38 patients in G1 tested negative for 

HCV-Ags, indicating 100% specificity of our HCV-Ags EIA. All 

85 patients with acute (G2, n=2) or chronic (G3, n=83) HCV 

infection (genotypes 1-6), and serum HCV RNA ranged from 

1,124 to 14,400,000 IU/mL quantified using PCR, tested positive 

for HCV-Ags with or without serum sample denaturation, 

indicating 100% sensitivity of the HCV-Ags EIA. The mean 

coefficient of variation (CVs) was 9.8% and 6.4%, respectively, 

for intra- precision assay and inter-assay reproducibility. 

In 2 cases with acute HCV infection, HCV-Ags were detectable 

59-65 days before anti-HCV test became positive. To further 

assess sensitivity, serum samples with low HCV RNA levels 

(214-1160 IU/mL) were serially diluted and tested. The lower 

limits of detection of the HCV-Ags assay using denatured or 

undenatured sera were equivalent to serum HCV RNA levels 

of 150 IU/mL and 250 IU/mL, respectively. Using denatured 

serum samples, 6/15 patients in G4 tested positive; however, 

none tested positive for HCV-Ags using undenatured serum 

samples. Additional studies showed that denaturation of these 

serum samples released HCV-Ags sequestered in HCV-ICs in 

these patients that may result in false positivity for active HCV 

infection. CONCLUSIONS. The highly specific and sensitive 

HCV-Ags EIA has a lower limit of detection equivalent to serum


HCV RNA levels of 150-250 IU/mL. Testing undenatured, but 

not denatured serum samples reliably differentiated active from 

past HCV infection and accomplished screening and diagnosis 

of HCV infection in one step. 

Disclosures: 

The following authors have nothing to disclose: Ke-Qin Hu, Wei Cui 

86 

Hepatitis C Virus (HCV) Mortality Patterns in the British 

Columbia Hepatitis Testers Cohort (BC-HTC) 

Mel Krajden 1,2 , Amanda Yu 1 , Darrel Cook 1 , Margot E. Kuo 1 , 

Maria Alvarez 1 , Mark Tyndall 3,2 , Naveed Z. Janjua 1,2 ; 1 Clinical 

Prevention Services, BC Centre for Disease Control, Vancouver, 

BC, Canada; 2 University of British Columbia, Vancouver, BC, Canada; 

3 BC Centre for Disease Control, Vancouver, BC, Canada 

Objective: To describe the population-level impacts of HCV 

acquisition risks vs. chronic infection on mortality for HCV 

negative and positive testers. Methods: The British Columbia 

Hepatitis Testers Cohort (BC-HTC) includes 1,135,947 individuals 

tested for HCV or reported to public health as a HCV 

case from 1990-2013, linked to their corresponding healthcare 

administrative data. Cohort entry was delayed one year 

for each individual and deaths within one year of HCV diagnosis 

were excluded, i.e., one year lagging. We computed 

age-adjusted annual all-cause, liver- and drug-related mortality 

rates/100,000 population and age-, sex- and year-adjusted 

standardized mortality ratios (SMRs) for: 1) seroconverters, 

those with known HCV acquisition timeframes and risk activities; 

2) anti-HCV positive on initial testing, the majority of whom 

were chronically infected and most of whom no longer engage 

in acquisition risk activities; and 3) HCV negatives. Results: 

Of 1,028,227 individuals included in this analysis, 64,876 

(6.3%) were HCV positive. Overall, 16.3% (10,572/64,876) 

of HCV positive vs. 6.4% (61,623/963,351) of HCV negative 

individuals died. For HCV positives, age-adjusted allcause 

mortality increased from 0.11/100,000 in 1993 to 

15.0 in 2005 and stayed in this range thereafter. Liver-related 

mortality increased from 0.03/100,000 in 1993 to 2.5 in 

2005 to 4.7 in 2013. Drug-related mortality increased from 

0.03/100,000 in 1993, peaked at 3.2 in 2005 and declined 

to 1.3 by 2013. All-cause, liver- and drug-related mortality 

rates were consistently higher for males vs. females. All-cause 

mortality for seroconverters was significantly higher than those 

with chronic HCV and HCV negatives (SMRs: 4.8, 3.1 and 

1.0). Liver-related mortality was significantly lower for seroconverters 

than for chronic HCV and was lowest for HCV negatives 

(SMRs: 10.4, 17.0 and 1.4). Similarly, liver cancer mortality 

was significantly lower for seroconverters than for chronic HCV 

and was lowest for HCV negatives (SMRs: 3.8, 17.5 and 1.3). 

However, drug-related mortality was highest for seroconverters 

compared to chronic HCV and HCV negatives (SMRs: 17.0, 

11.4 and 0.9). Conclusions: The BC-HTC enables comprehensive 

assessment of the mortality impacts of HCV infection and 

creates an opportunity to differentiate acquisition risk mortality 

from the impacts of chronic infection. These data illustrate that 

the excess mortality related to HCV acquisition risks is distinct 

from viral sequelae for a significant proportion of the HCV 

infected population. Therefore antiviral treatment on its own 

will not optimize mortality reductions in the affected populations. 

Disclosures: 

Mel Krajden - Grant/Research Support: Roche, Merck, Siemens, Boerhinger 

Ingelheim, Hologic 

The following authors have nothing to disclose: Amanda Yu, Darrel Cook, Margot 

E. Kuo, Maria Alvarez, Mark Tyndall, Naveed Z. Janjua 

87 

The Association of Sustained Virological Response and 

All-cause Mortality After Interferon-based Therapy for 

Chronic Hepatitis C (HCV) in a Large U.S. Community-based 

Health Care Delivery System 

Lisa M. Nyberg 1 , Xia Li 1 , Su-Jau Yang 1 , Kevin Chiang 1 , T. Craig 

Cheetham 1 , Susan Caparosa 1 , Jose R. Pio 1 , Zobair M. Younossi 2 , 

Anders H. Nyberg 1 ; 1 Kaiser Permanente, San Diego, CA; 2 Department 

of Medicine, Inova Fairfax Medical Campus, Fairfax, VA 

Background: Previous studies performed at tertiary care centers 

have shown that sustained virological response (SVR) to interferon-based 

therapy for HCV is associated with a reduction 

of liver-related and all-cause mortality (1-2). The aim of this 

study was to determine if SVR to interferon-based therapy in 

a community-based health care setting is associated with a 

reduction in all-cause mortality and to examine for differences 

in those with and without cirrhosis. Methods: A retrospective 

cohort study at Kaiser Permanente, Southern California (KPSC), 

a large community-based integrated health care system including 

3.5 million members. Inclusion criteria: a diagnosis code 

and/or positive lab test for HCV RNA (index date) 1/1/02- 

12/31/13; age ≥ 18 years at index date, ≥ 12 months continuous 

membership before and after index date. Exclusion 

criteria: HCV diagnosis after 1/1/13 and liver transplant or 

HCC on or before index date. SVR was determined for patients 

treated for HCV with interferon-based therapy and stratified for 

cirrhosis vs non-cirrhosis. Mortality data were obtained from the 

California Department of Public Health Vital Statistics of California 

Report. Results: Total study cohort: 24,968 with chronic 

HCV; of those 10,449 (42%) had cirrhosis. Overall mortality 

during the study period was 18.5% (4,608/24,968). Mortality 

among those with cirrhosis was 33.2% (3,470/10,449) vs 

7.8% (1,138/14,519) among those without cirrhosis. Among 

patients treated for HCV, 45.1% (2348/5203) achieved SVR; 

mortality for those who achieved SVR was 5.4% (127/2348) 

vs 18.9% (540/2855) for those without SVR. Mortality in 

treated patients, with and without cirrhosis, is shown in Table 

1. Conclusions: An approximate 3-fold reduction in all-cause 

mortality is seen in patients with HCV who are treated and 

achieve SVR compared to those without SVR. Cirrhotics who 

are treated and achieve SVR have a > 3-fold reduction in mortality 

compared to the mortality in treated cirrhotics who do not 

achieve SVR. New, more potent, HCV treatment regimens with 

higher SVR have the potential of significantly reducing future 

mortality due to HCV. Further study is ongoing with matching 

for severity of liver disease to more specifically examine the 

effect of SVR on mortality compared to that due to the natural 

history of HCV/cirrhosis. 1. van der Meer, et al. JAMA 2012 

Dec26;308(24):2584-93. 2. Berenguer, et al. Hepatology 

2009 Aug;50(2):407-13 

Mortality in HCV patients treated with interferon-based therapy. 

Cirrhosis: N=2603; No Cirrhosis: N=2600 

Disclosures: 

Lisa M. Nyberg - Grant/Research Support: Merck, Gilead, Abbvie 

T. Craig Cheetham - Grant/Research Support: Gilead, BMS 

Zobair M. Younossi - Advisory Committees or Review Panels: Salix, Janssen, 

Vertex; Consulting: Gilead, Enterome, Coneatus 

The following authors have nothing to disclose: Xia Li, Su-Jau Yang, Kevin Chiang, 

Susan Caparosa, Jose R. Pio, Anders H. Nyberg


88 

Increasing Prevalence of Cirrhosis among US Adults 

with Chronic Hepatitis C Virus Infection: Results from 

NHANES 1988-1994 and 1999-2012 

Prowpanga Udompap, Ajitha Mannalithara, Nae-Yun Heo, Donghee 

Kim, W. Ray Kim; Division of Gastroenterology and Hepatology, 

Stanford University School of Medicine, Stanford, CA 

Background & Aims: Hepatitis C virus(HCV) is a major cause 

of end stage liver disease and hepatocellular carcinoma worldwide. 

While cirrhosis underlies these complications, the prevalence 

of HCV cirrhosis in the US remains unknown. We aim 

to determine the prevalence of advanced fibrosis/cirrhosis and 

to identify factors associated with advanced fibrosis/cirrhosis 

in people with chronic hepatitis C in the US population. Methods: 

Using the National Health and Nutrition Examination Survey(NHANES) 

data, we identified participants with detectable 

serum HCV RNA and assessed liver fibrosis using validated 

surrogate indicators, including APRI and FIB-4 scores. The prevalence 

of cirrhosis was determined for survey participants for 

Era 1 (1988-94), Era 2 (1999-2006) and Era 3 (2007-12). 

Results: Out of 52,644 participants with age ≥20, 736 (1.4%) 

had HCV. Based on APRI score, 6.6% (95%CI:2.2-11.0) of 

US adults with HCV infection in Era 1, 7.6% (95%CI:3.4- 

11.8) in Era 2 and 17.0% (95%CI:8.0-26.0) in Era 3 were 

estimated to have cirrhosis (Ishak stage 5-6). The prevalence 

of advanced fibrosis (Ishak stage 4-6) based on FIB-4 was 

8.6%, 10.1%, and 16.0% for Eras 1, 2, and 3, respectively. 

These data project to 370,500 Americans with cirrhosis and 

347,800 with advanced fibrosis in the most recent era. The 

higher prevalence of cirrhosis (determined by APRI) in the 

recent era was associated with increasing age (OR=1.04, 

95%CI:1.02-1.07), diabetes (OR=2.33, 95%CI:1.01-5.40) 

and obesity (OR=2.96, 95%CI:1.15-7.57). When FIB-4 score 

was used, advanced fibrosis was associated with increasing 

age (OR=1.08, 95%CI:1.05-1.11) and diabetes (OR=3.37, 

95%CI:1.24-9.5). Conclusion: Nearly one in five US adults 

with HCV infection have cirrhosis. The prevalence of cirrhosis 

is rising over time, which is in part attributable to the increasing 

age of the birth cohort with HCV infection. In addition, comorbidities 

such as diabetes and obesity accelerate liver fibrosis. 

These data further underscore the current recommendations 

for HCV screening in asymptomatic individuals and highlights 

the need for systematic assessment for liver fibrosis and comprehensive 

medical management in those with HCV infection. 

Characteristics of participants 

89 

Rectal Shedding of HCV in HCV/HIV Co-infected Men 

Andrew L. Foster 1,2 , Michael Gaisa 1 , Rosanne M. Hijdra 1,3 , Karen 

B. Jacobson 1 , Samuel Turner 1,2 , Tristan Morey 1,2 , Daniel S. Fierer 1 ; 

1 Infectious Diseases, Mount Sinai School of Medicine, New York, 

NY; 2 James Cook University, Cairns, QLD, Australia; 3 Amsterdam 

Medical Center, Amsterdam, Netherlands 

Introduction An epidemic of hepatitis C virus (HCV) infection 

is occurring among HIV-infected men who have sex with men 

(MSM). Epidemiological studies suggest that sexual transmission 

is fueling the epidemic. Blood and semen have been considered 

as potential mechanisms of transmission, but there are 

still transmission circumstances that remain unexplained. We 

hypothesized that HCV may be shed into rectal fluid of HCV/ 

HIV co-infected MSM. Methods Written informed consent was 

obtained from 45 HIV-infected MSM with HCV infection and 

paired blood and rectal fluid specimens were obtained. Rectal 

fluid was collected using a moistened polyester-tipped swab 

that was gently inserted into the rectum. The swab was placed 

into transport medium, vortexed, and the supernatant analysed 

for HCV using COBAS AMPLICOR (Roche Diagnostics), lower 

limit of quantification 43 IU/mL, lower limit of detection 7 IU/ 

mL. The experimentally-determined efficiency of HCV absorption 

to and elution from the swabs was used to calculate the 

rectal HCV viral load (VL). Rectal sexually-transmitted infection 

(STI) and blood syphilis testing were also performed. Results 

Visual inspection of the swabs showed no visible blood on 

any. HCV was detected in 20 (47%) of 43 specimens. Among 

those samples with HCV detected, the median rectal VL was 

2.92 log 10 

IU/mL (IQR 2.92, 4.27). Rectal HCV detection was 

associated with serum HCV VL >5 log 10 

IU/mL (p=0.011), 

and there was a high correlation between the magnitude of 

blood VL and rectal HCV VL (correlation coefficient 0.688, 

p


patients in care with hepatitis C virus infection (HCV) will 

achieve this status in the near future. However, the long term 

prognosis in terms of risk or predictors of developing hepatocellular 

carcinoma (HCC) among patients with SVR remains 

unclear. Methods: A retrospective cohort study using data from 

the national VA HCV Clinical Case Registry in patients with 

HCV diagnosed (positive HCV RNA) between 10/1999 and 

1/2009 who had at-least one year of follow up in the VA. 

HCV treatment (pegylated interferon with or without ribavirin) 

and SVR status (RNA test negative at least 12 weeks after 

the end of treatment) were ascertained. Cox proportional hazards 

models were used to examine the effect of demographic, 

virological and clinical factors on time to HCC among those 

who achieved SVR through end of 2011 while adjusting for 

medical comorbidity. We excluded those with HCC diagnostic 

codes before SVR date. Results: We had 33,005 HCV-infected 

individuals who received HCV treatment during the study timeframe. 

Of these, 10,817 patients achieved SVR and 10765 

had no HCC before SVR. Their mean age was 53.1 (sd, 6.3), 

12.4% were 60 years or older, and 12.9% were Black, 64.4% 

non-Hispanic white, and 3.4% Hispanic. Majority (95.3%) 

were males; and 53.6% had genotype 1, 25.0% genotype 

2, 13.5% genotype 3, 0.7% genotype 4, and 7.2% had no 

genotype test available. A total of 124 patients developed 

HCC during a total follow of 38,394 person year follow up 

(median duration of follow up after SVR was 2.7 years) at 

an incidence rate of 0.32% per year. In the cohort with SVR, 

patients 60 years and older (HR=6.59 c/w younger age), 

Hispanics (HR=2.16, 1.03-4.50 c/w non-Hispanic whites) and 

those with cirrhosis (HR=5.58, 3.21 - 9.68) or diabetes (1.82 

(0.92 - 3.61) were at an increased risk of developing HCC 

compared to their counterparts. Conclusions: The risk of HCC 

after HCV cure remains elevated (0.3% per year). Older age 

at the time of SVR, Hispanic ethnicity, and presence of cirrhosis 

and diabetes are associated with an increased HCC risk 

among those who achieve SVR. 

Disclosures: 

Hashem B. El-Serag - Grant/Research Support: WAKO, Gilead 

The following authors have nothing to disclose: Peter Richardson, Fasiha Kanwal 

91 

Prevalence of Pre-Treatment NS5A Resistance Associated 

Variants in Genotype 1 Patients Across Different 

Regions Using Deep Sequencing and Effect on Treatment 

Outcome with LDV/SOF 

Stefan Zeuzem 2 , Masashi Mizokami 3 , Stephen Pianko 4 , Alessandra 

Mangia 5 , Kwang-Hyub Han 6 , Ross Martin 1 , Evguenia 

S. Svarovskaia 1 , Hadas Dvory-Sobol 1 , Brian Doehle 1 , Phillip S. 

Pang 1 , Steven J. Knox 1 , John G. McHutchison 1 , Diana M. Brainard 

1 , Michael D. Miller 1 , Hongmei Mo 1 , Wan-Long Chuang 7 , 

Ira M. Jacobson 8 , Gregory Dore 9 , Mark S. Sulkowski 10 ; 1 Gilead 

Sciences Inc, Foster City, CA; 2 Department of Internal Medicine, 

J.W. Goethe University Hospital, Frankfurt am Main, Germany; 

3 National Center for Global Health and Medicine, Tokyo, Japan; 

4 Department of Gastroenterology, Monash Medical Centre, Victoria, 

VIC, Australia; 5 Liver Unit, IRCCS-Ospedale Casa Sollievo 

della Sofferenza, San Giovanni Rotondo, Italy; 6 Department of 

Internal Medicine, Yonsei University College of Medicine, Seoul, 

Korea (the Republic of); 7 Internal Medicine, Kaohsiung Medical 

University Hospital, Kaohsiung, Taiwan; 8 Medicine, Mt. Sinai Beth 

Israel, New York, NY; 9 Kirby Institute, UNSW Australia, Sydney, 

NSW, Australia; 10 School of Medicine, Johns Hopkins University, 

Baltimore, MD 

Background: The efficacy of some HCV regimens is influenced 

by the presence of certain NS5A resistance associated variants 

(RAVs). To investigate if the pretreatment prevalence of 

NS5A RAVs in genotypes (GT) 1a and 1b varied by country, 

race or ethnicity, comprehensive analyses were performed 

using deep sequencing of NS5A from more than 5000 patients 

from 17 countries. Methods: NS5A deep sequencing analysis 

with a 1% cut-off was performed. GT1a RAVs were defined 

as K24G/N/R, M28A/G/T/V, Q30H/G/R/E/K, L31any, 

P32L, S38F, H58D, A92K/T, Y93any, and GT1b RAVs were 

L31any, P32L, P58D, A92K, Y93any. Results: Pretreatment 

samples from 5046 patients were analyzed. Prevalence of 

NS5A RAVs was similar between the different regions for both 

GT1a and GT1b HCV infected patients (Table). In addition, no 

significant differences were observed in NS5A RAV prevalence 

between patients with different race or ethnicity. In the subset of 

patients who were treated with LDV/SOF for 12 weeks, SVR12 

rates were similar in GT1b patients with and without pretreatment 

NS5A RAVs across all regions. In GT1a patients with 

pretreatment RAVs, SVR12 rates in North America were 91% 

(75/82) compared to 98% without NS5A RAVs. All seven 

GT1a patients who relapsed had pretreatment NS5A RAVs 

conferring >1000-fold reduced susceptibility to LDV (H58D, 

Y93H/N/F or multiple RAV combinations). However, 18 GT1a 

patients with similar RAVs conferring >1000-fold reduced 

susceptibility achieved SVR12 after LDV/SOF for 12 weeks. 

Conclusions: No significant difference in prevalence of NS5A 

pretreatment RAVs was observed between different regions, 

races or ethnicities. Overall, high SVR12 rates (91-100%) were 

observed in patients with or without NS5A RAVs with LDV/ 

SOF. In GT1a patients, lower SVR rate (72%) was observed 

in patients with pretreatment NS5A RAVs conferring high level 

(>1000-fold) resistance to NS5A inhibitors. 

NA: Too few patients treated with LDV/SOF 

Disclosures: 



Stephen Pianko - Advisory Committees or Review Panels: Roche, Novartis, GIL- 

EAD, Roche, Novartis; Consulting: GILEAD; Speaking and Teaching: JANSSEN 

Alessandra Mangia - Advisory Committees or Review Panels: ROCHE, Janssen, 

MSD, ROCHE, Janssen, MSD, Boheringer ; Consulting: Gilead; Grant/Research 

Support: Shering-Plough, Shering-Plough 

Ross Martin - Employment: Gilead Sciences 

Evguenia S. Svarovskaia - Employment: Gilead Sciences Inc; Stock Shareholder: 

Gilead Sciences Inc 

Hadas Dvory-Sobol - Employment: Gilead Sciences; Stock Shareholder: Gilead 

Sciences 

Brian Doehle - Employment: Gilead Sciences 

Phillip S. Pang - Employment: Gilead Sciences; Stock Shareholder: Gilead Sciences 

Steven J. Knox - Employment: Gilead Sciences 

John G. McHutchison - Employment: Gilead Sciences; Stock Shareholder: Gilead 

Sciences 

Diana M. Brainard - Employment: Gilead Sciences; Stock Shareholder: Gilead 

Sciences 

Michael D. Miller - Employment: Gilead Sciences, Inc.; Stock Shareholder: Gilead 

Sciences, Inc. 

Hongmei Mo - Employment: Gilead Science Inc


Wan-Long Chuang - Advisory Committees or Review Panels: Gilead, Abbvie; 

Speaking and Teaching: BMS, Roche, MSD 

Ira M. Jacobson - Consulting: AbbVie, Achillion, Alnylam, Bristol Myers Squibb, 

Enanta, Gilead, Janssen, Merck; Grant/Research Support: AbbVie, Bristol Myers 

Squibb, Gilead, Janssen, Merck, Tobira; Speaking and Teaching: AbbVie, Bristol 

Myers Squibb, Gilead, Janssen 




Mark S. Sulkowski - Advisory Committees or Review Panels: Merck, AbbVie, 

Janssen, Gilead, BMS; Grant/Research Support: Merck, AbbVie, Janssen, Gilead, 

BMS 

The following authors have nothing to disclose: Masashi Mizokami, Kwang-Hyub 

Han 

92 

Highly Successful Retreatment with Ledipasvir (LDV) and 

Sofosbuvir (SOF) in HCV GT-1 Patients Who Failed Initial 

Short Course Therapy with Combination DAA Regimens 

(NIH SYNERGY Trial) 

Eleanor Wilson 1,2 , Sarah Kattakuzhy 1 , Zayani Sims 2 , Mary 

McLaughlin 3 , Angie Price 1 , Hongmei Mo 4 , Anu Osinusi 4 , Henry 

Masur 2 , Anita Kohli 5 , Shyam Kottilil 1 ; 1 Infectious Disease, Institute 

for Human Virology/University of Maryland School of Medicine, 

Rockville, MD; 2 Critical Care Medicine Department, National Institutes 

of Health, Bethesda, MD; 3 National Institute of Allergy and 

Infectious Diseases, National Institutes of Health, Bethesda, MD; 

4 Gilead Sciences, Foster City, CA; 5 Hepatology, St. Joseph’s Hospital 

and Medical Center, Phoenix, AZ 

Introduction: Directly-acting antivirals (DAAs) dramatically 

improved HCV therapy but retreatment options for patients 

who have failed therapy with combination DAAs have not 

been studied. Our study aim is to determine if HCV genotype-1 

patients who failed short course combination therapy with 3 

or 4 DAAs can be successfully treated with 12 weeks of LDV/ 

SOF. Material and Methods: In this single-center, open-label, 

phase 2a trial, HCV mono-infected persons with early stage 

(F0-F2) liver fibrosis and previous exposure to combination 

DAA therapy only (LDV/SOF with GS-9451 +/- GS-9669) 

were eligible to enroll and receive 12 weeks of LDV/SOF. 

HCV RNA was measured with the Abbott assay, lower level 

of quantitation (LLOQ) 12 IU/ml. The primary endpoint was 

defined as HCV viral load (VL) < LLOQ 12 weeks after end 

of therapy (SVR12). Deep sequencing of NS5B and NS5A 

regions was performed at baseline by Illumina next generation 

sequencing technology. Results: The study enrolled 34 persons, 

and 32 (94%) completed therapy with 12 weeks of LDV/ 

SOF. Two patients withdrew consent after Day 0. Participants 

were predominantly male (82%) and black (85%), median 

age 60.5 years (IQR 57.0 – 63.8) and BMI 26.8 kg/m 2 (IQR 

25.3 – 29.3). Baseline HCV VL was 1.3 x 10^6 IU/mL (IQR 

5.8x10^5 – 3.9x10^6), 73.5% (25/34) were infected with 

HCV genotype 1a, and median Metavir fibrosis stage was 

1. Time from relapse to retreatment was 22 weeks (IQR 18 – 

23). SVR12 rates were 91% (31/34; ITT) and 97% (31/32, 

per protocol). For SVR rates by original treatment group (per 

protocol), see Figure. At baseline, 28/33 patients (85%) had 

resistance-associated variants (RAVs) consistent with >25 fold 

resistance in NS5A. Of all patients completing therapy, 1 

patient with NS5A RAVs relapsed. Conclusions: For the first 

time, we demonstrate a high SVR rate following retreatment 

with DAAs in patients who have previously failed DAA-only 

therapy. 

Disclosures: 

Hongmei Mo - Employment: Gilead Science Inc 

Anu Osinusi - Employment: gilead sciences 

The following authors have nothing to disclose: Eleanor Wilson, Sarah Kattakuzhy, 

Zayani Sims, Mary McLaughlin, Angie Price, Henry Masur, Anita Kohli, 

Shyam Kottilil 

93 

Effectiveness of Ledipasvir/Sofosbuvir in Treatment 

Naïve Genotype 1 Patients Treated in Routine Medical 

Practice 

Lisa I. Backus 1,2 , Pamela S. Belperio 1 , Troy Shahoumian 1 , Timothy 

P. Loomis 1 , Larry A. Mole 2 ; 1 Office of Public Health/Population 

Health, Veterans Affairs Palo Alto Health Care System, Palo Alto, 

CA; 2 Department of Medicine, VA Palo Alto Healthcare System, 

Palo Alto, CA 

Aim: Assess the effectiveness of ledipasvir/sofosbuvir±ribavirin 

(LDV/SOF±RBV) in treatment naïve genotype 1 (GT1) hepatitis 

C virus (HCV)-infected veterans treated in routine medical practice. 

Methods: This observational, intent-to-treat cohort analysis 

used the Veterans Affairs’ Clinical Case Registry to identify all 

treatment naïve GT1 HCV-infected veterans initiating 8 or 12 

weeks of LDV/SOF±RBV by 31 December 2014. Patients were 

excluded for liver transplantation or baseline HCV RNA


ics with baseline HCV RNA


95 

Improving liver function and delisting of patients awaiting 

liver transplantation for HCV cirrhosis: do we ask 

too much to DAA? 

Audrey Coilly 1,2 , Georges-Philippe Pageaux 22 , Pauline Houssel-Debry 

7 , Christophe Duvoux 3 , Sylvie Radenne 6 , Victor de 

Ledinghen 15 , Danielle Botta-Fridlund 11 , Anaïs Vallet-Pichard 13 , 

Rodolphe Anty 9 , Vincent Di Martino 5 , Filomena Conti 10 , Marie Line 

Debette-Gratien 20 , Laurent Alric 16 , Armando Abergel 17 , Camille 

Besch 19 , Helene Montialoux 18 , Pascal Lebray 10 , Sebastien Dharancy 

4 , Francois Durand 12 , Louis d’Alteroche 21 , Florian Charier 8 , 

Olivier Chazouillères 14 , Jérôme Dumortier 24 , Vincent Leroy 23 , Jean- 

Charles Duclos-Vallee 1,2 ; 1 Centre Hepato-Biliaire, AP-HP Hopital 

Paul-Brousse, Villejuif, France; 2 Unit 1193, INSERM, Villejuif, 

France; 3 AP-HP Hopital Henri Mondor, Creteil, France; 4 CHRU 

de Lille, Lille, France; 5 CHU Jean Minjoz, Besançon, France; 6 HCL 

- Hopital Croix Rousse, Lyon, France; 7 CHU de Rennes - Hopital 

Pontchaillou, Renes, France; 8 CHU de Poitiers, Poitiers, France; 

9 CHU de Nice, Nice, France; 10 AP-HP Hopital Pitie-Salpetrière, 

Paris, France; 11 AP-HM Hopital La Conception, Marseille, France; 

12 AP-HP Hopital Beaujon, Clichy, France; 13 AP-HP Hopital Cochin, 

Paris, France; 14 AP-HP Hopital Saint-Antoine, Paris, France; 15 CHU 

de Bordeaux - Hopital Haut-Leveque, Pessac, France; 16 CHU de 

Toulouse - Hopital Purpan, Toulouse, France; 17 CHU de Clermont-Ferrand, 

Clermont-Ferrand, France; 18 CHU de Rouen, Rouen, 

France; 19 CHU de Strasbourg - Hopital Hautepierre, Strasbourg, 

France; 20 CHU de Limoges, Limoges, France; 21 CHU de Tours 

- Hopital Trousseau, Tours, France; 22 CHU de Montpellier - Hopital 

Saint-Eloi, Montpellier, France; 23 CHU de Grenoble - Hopital 

Michallon, Grenoble, France; 24 HCL - Hopital Edouard Herriot, 

Lyon, France 

Background: Combinations of DAA have shown excellent 

results to treat HCV-infection in cirrhotic patients (pts). But some 

issues remain unresolved regarding efficacy in pts awaiting 

liver transplantation (LT) and impact on access to LT. Methods: 

This cohort study enrolled 151 pts registered in 23 centers 

on French LT list (male: 79%, age 56±7 years), treated with 

sofosbuvir ± ribavirin (n=84) ± daclatasvir (n=90) or simeprevir 

(n=9) or ledipasvir (n=17). Meantime between listing and 

starting therapy was 23±57 weeks (wks). All pts were cirrhotic. 

LT indication was HCC in 85(56%) pts. 112(74%) pts were 

treatment experienced including 46% of non-responders, failures 

to 1rst generation protease inhibitors in 32 or sofosbuvir 

in 16 pts. Majority of pts were genotype 1 (56%) and 3 (24%). 

Results: Mean follow-up was 44±19 wks [12-79]. Mean baseline 

MELD score (MELD), platelet count, bilirubin and albumin 

levels were 10±5 [6-32], 89±50 G/L[23-347], 39±42μmol/L 

[5-405], 33±7 g/L [15-47]. 73 pts (48%) were decompensated 

(bilirubin > 50μmol/L in 34(23%) ± ascites in 53(35%) 

± hepatic encephalopathy (HE) in 29(19%)). Indetectability 

of HCV RNA was obtained at 7±4 wks [1-24]. Among 117 

pts, 103(88%) achieved SVR12. Among 73 decompensated 

pts, a complete response (normal bilirubin level, no ascites, 

no HE) was observed in 14(19%) after 12 wks and 31(42%) 

at 12 wks post-treatment. When achieving SVR12, ascites or 

HE persisted in 12/44 (27%) and 7/14 (50%). Variations of 

MELD are shown in Table 1. Among patients with baseline 

MELD score≥20, 83% still an indication of LT at the end of treatment 

(MELD≥15). 45(30%) pts were transplanted. Ten (6%) pts 

were delisted because of liver function improvement. A serious 

adverse event occurred in 24(19%) pts, mainly liver events 

(37%), anemia (25%) and sepsis (25%). Conclusion: Among 

151 cirrhotic pts awaiting LT treated with IFN-free regimens, 

88% achieved SVR12. Only 6% of pts were delisted due to 

liver function improvement. A complete clinical and biological 

response was observed in 42%, raising hopes that more pts 

could be delisted. However, major clinical improvement was 

observed in a minority of patients especially when MELD score 

was ≥20, suggesting that antiviral treatment might be postponed 

after LT in this population. Final results will be provided 

in a larger population. 

Variations of MELD score in 66 pts with decompensated condition 

from baseline to the end of treatment 

Disclosures: 

Audrey Coilly - Consulting: Novartis, Astellas, Janssen, Bristol-Myers-Squibb, 

Merck Sharp & Dohme, Gilead, Roche 

Georges-Philippe Pageaux - Advisory Committees or Review Panels: Roche, 

Roche, Roche, Roche; Board Membership: Astellas, Astellas, Astellas, Astellas 

Pauline Houssel-Debry - Speaking and Teaching: NOVARTIS, ASTELLAS, GILEAD 

Christophe Duvoux - Advisory Committees or Review Panels: Novartis, Roche, 

Novartis, Roche, Novartis, Roche, Novartis, Roche; Speaking and Teaching: 

Astellas, Astellas, Astellas, Astellas 

Victor de Ledinghen - Board Membership: Janssen, Gilead, BMS, Abbvie; Speaking 

and Teaching: AbbVie, Merck, BMS, Gilead 

Danielle Botta-Fridlund - Consulting: GILEAD, ABBVIE, BMS, MSD 

Anaïs Vallet-Pichard - Independent Contractor: Schering Plough, Gilead, BMS, 

Roche 

Vincent Di Martino - Advisory Committees or Review Panels: Gilead, France, 

Abbvie, BMS France; Board Membership: MSD France; Consulting: Gilead, 

France; Speaking and Teaching: Janssen, BMS France, Gilead France 

Laurent Alric - Board Membership: Schering Plough, Schering Plough, Schering 

Plough, Schering Plough; Consulting: MSD; Speaking and Teaching: Roches, 

BMS, Gilead, Roches, BMS, Gilead, Roches, BMS, Gilead, Roches, BMS, Gilead, 

MSD, Abbvie 

Armando Abergel - Consulting: gilead, msd, bms; Speaking and Teaching: abbvie 

Pascal Lebray - Grant/Research Support: Schering-Plough, Schering-Plough, Schering-Plough, 

Schering-Plough; Speaking and Teaching: Gilead, Gilead, Gilead, 

Gilead 

Sebastien Dharancy - Advisory Committees or Review Panels: CHIESI; Board 

Membership: NOVARTIS; Speaking and Teaching: ASTELLAS 

Francois Durand - Advisory Committees or Review Panels: Astellas, Novartis, 

BMS; Speaking and Teaching: Gilead 


Jérôme Dumortier - Board Membership: Novartis, Astellas, Roche; Consulting: 

Novartis; Grant/Research Support: Novartis, Astellas, Roche, MSD, GSK 

Vincent Leroy - Board Membership: Abbvie, BMS, Gilead; Consulting: Janssen, 

MSD; Speaking and Teaching: Abbvie, BMS, Gilead, Janssen, MSD 

The following authors have nothing to disclose: Sylvie Radenne, Rodolphe Anty, 

Filomena Conti, Marie Line Debette-Gratien, Camille Besch, Helene Montialoux, 

Louis d’Alteroche, Florian Charier, Jean-Charles Duclos-Vallee 

96 

Detecting Drug-Induced Liver Injury in Patients with 

Decompensated Chronic Hepatitis C: A Review of the 

SOLAR-1 and SOLAR-2 Studies 

Andrew J. Muir 2 , Michael R. Charlton 5 , Phillip S. Pang 1 , Luisa 

M. Stamm 1 , Diana M. Brainard 1 , John G. McHutchison 1 , Nezam 

H. Afdhal 3 , Paul B. Watkins 4 ; 1 Gilead Sciences, Foster City, CA; 

2 Duke University Medical Center, Durham, NC; 3 Division of Gastroenterology, 

BIDMC, Boston, MA; 4 The Hamner-UNC Institute for 

Drug Safety Sciences, Chapel Hill, NC; 5 Intermountain Medical 

Center, Murray, UT 

Background: The identification of drug-induced liver injury 

(DILI) is challenging in patients with underlying liver disease, 

particularly in the setting of decompensated cirrhosis which has 

recently become a target population for novel therapies. Traditional 

laboratory criteria (e.g., elevation in serum ALT > 5x 

upper limit of normal) may not be appropriate for patients with


advanced liver disease. The aim of this analysis was to develop 

new criteria for identifying DILI associated with direct-acting 

antivirals in end-stage liver disease patients. Methods: The 

SOLAR-1 and SOLAR-2 studies assessed the safety and efficacy 

of ledipasvir/sofosbuvir plus ribavirin (LDV/SOF+RBV) for 12 

or 24 weeks in patients with advanced liver disease pre- or 

post-liver transplant. Laboratory data, including total and direct 

bilirubin and ALT and AST relative to baseline, from patients 

with CPT B and C cirrhosis (N=328) were used to determine 

criteria to screen for possible cases of DILI. Cases of interest, 

including clinical outcome data, were further evaluated for DILI 

by an expert panel. DILI was classified as possible or unlikely, 

where unlikely required the identification of a clear non-drug 

related etiology of the decompensation or laboratory abnormalities. 

Results: Overall, on-treatment elevations above the 

upper limit of normal were common for total bilirubin (90%, 

n=295), ALT (48%, n=157) and AST (90%, n=294). Further 

analysis of total bilirubin identified only 11 of 295 patients in 

whom the direct bilirubin had a treatment-emergent elevation of 

greater than 1 mg/dL. Adjudication of these 11 cases revealed 

1 possible case of DILI. The use of ALT and AST criteria, including 

a cutoff of 5x nadir, did not identify further cases of interest 

beyond those identified by the >1 mg/dL increase in direct 

bilirubin. Median ALT and AST at baseline were 62 IU/ml and 

100 IU/ml, respectively; only 3 subjects reached a maximum 

ALT or AST > 2x baseline. ALT and AST improved (on-treatment 

ALT maximum < baseline) in 91% and 92% of patients, 

respectively. Conclusions: In patients with CPT B or C cirrhosis 

and HCV treated with LDV/SOF+RBV, elevation from baseline 

in direct bilirubin of > 1mg/dL was a clinically sensitive and 

conservative cutoff that identified patients who should be further 

evaluated for the possibility of DILI. No additional possible 

cases of DILI were identified by using ALT or AST criteria. An 

adjudication of all biochemical and clinical events of interest 

(including deaths and liver transplantations) of all patients in 

the SOLAR-1 and SOLAR-2 studies will be presented. 

Disclosures: 

Andrew J. Muir - Advisory Committees or Review Panels: BMS, Gilead, Janssen, 

Merck; Consulting: Theravance; Grant/Research Support: Abbvie, Abbvie, BMS, 

Gilead, Janssen, Merck, Achillion, Lumena 

Michael R. Charlton - Grant/Research Support: GIlead Sciences, Merck, Janssen, 

AbbVie, Novartis 


Luisa M. Stamm - Employment: Gilead Sciences 


Sciences 


Sciences 

Nezam H. Afdhal - Advisory Committees or Review Panels: Trio Helath Care; 

Board Membership: Journal Viral hepatitis; Consulting: Merck, EchoSens, BMS, 

Achillion, GlaxoSmithKline, Springbank, Gilead, AbbVie; Grant/Research Support: 

Gilead; Stock Shareholder: Springbank 

Paul B. Watkins - Consulting: Abbott, Actelion, Boerringer-Ingelheim, Cempra, 

Alecra, Roche, Merck, Reservlogix, Intercept, Janssen, Novartis, Otsuka, Pfizer, 

Sanolfi, Takeda, UCB, Bristol-Myers Squibb, GSK 

the underlying mechanism driving apoptosis remains unclear. 

In this study we investigated the role of autophagy in regulating 

apoptosis in NAFLD. Methods and Results: Administration of 

palmitic acid (PA) to HepG2 cells increased Annexin V-positive/7-AAD-negative 

cells and induced apoptosis from 8 hours, 

which was found to be mediated by activation of IRE1-JNK and 

PERK-CHOP pathways. PA increased the expression level of 

LC3-II and decreased the autophagic flux index, which can be 

measured by monitoring LC3-II turnover using bafilomycin A1, 

from 3 hours. These findings suggest that PA inhibits the autophagic 

process after autophagosome formation. In PA-treated 

HepG2 cells, mTOR signaling, a negative regulator of autophagy, 

was inhibited and the expressions levels of Beclin1, 

Atg7 and Atg5 were unaltered. In contrast, the expression 

level of Rubicon, a negative regulator of autophagosome-lysosome 

fusion, was clearly upregulated by PA treatment from 

2 hours. siRNA-mediated Rubicon knockdown clearly abolished 

PA-induced inhibition of autophagic flux, reduced ER 

stress and ameliorated PA-induced apoptosis. While PA treatment 

did not change the levels of Rubicon mRNA expression, 

a pulse chase assay revealed that Rubicon protein stability 

was enhanced by PA treatment. Consistent with in vitro findings, 

mice on high fat diet (HFD) showed increased Rubicon 

expression from 1 month, autophagy inhibition from 2 months 

and increased apoptosis from 3 months in the liver. Electron 

microscopy revealed that isolation membranes and autophagosomes 

in the liver of mice fed HFD were increased, suggesting 

the inhibition of autophagy after autophagosome maturation. 

Hepatocyte-specific Rubicon knockout (KO) mice on HFD for 4 

months improved autophagy inhibition, ER stress and hepatocyte 

apoptosis compared with wild-type littermates. They also 

showed the significant reduction of liver volume to approximately 

normal level, which was accompanied by significant 

and substantial reduction of triglycerides in their livers. Finally, 

we investigated Rubicon expression levels in human liver samples 

by western blotting under approval of institutional ethics 

committee. Rubicon expression levels in steatotic livers were 

higher than those in non-steatotic livers. Conclusion: Increased 

expression of Rubicon induced by PA and HFD suppresses 

autophagic flux and promotes hepatocyte apoptosis by increasing 

ER stress. Enhancement of autophagy by inhibiting Rubicon 

may provide new approaches for treatment of NAFLD. 

Disclosures: 






The following authors have nothing to disclose: Satoshi Tanaka, Sadatsugu 

Sakane, Yasutoshi Nozaki, Yugo Kai, Tasuku Nakabori, Yoshinobu Saito, 

Ryotaro Sakamori, Tomohide Tatsumi 

97 

Enhanced expression of Rubicon inhibits autophagy and 

promotes apoptosis by increasing endoplasmic reticulum 

stress in nonalcoholic fatty liver disease 

Satoshi Tanaka, Hayato Hikita, Sadatsugu Sakane, Yasutoshi 

Nozaki, Yugo Kai, Tasuku Nakabori, Yoshinobu Saito, Ryotaro 

Sakamori, Tomohide Tatsumi, Tetsuo Takehara; Gastroenterology 

& Hepatology, Osaka University, Suita, Japan 

Background and Aim: Hepatocyte apoptosis is a characteristic 

feature of nonalcoholic fatty liver disease (NAFLD). However,


98 

In vivo reprogramming of myofibroblasts into hepatocytes 

as a therapy for liver fibrosis 

Milad Rezvani 1 , Regina Español-Suñer 1 , Yann Malato 1 , Laure 

Dumont 1 , Andrew A. Grimm 2 , Eike Kienle 3 , Julia Bindmann 1 , Ellen 

Wiedtke 3 , Syed J. Naqvi 1 , S. C. Derderian 4 , Robert F. Schwabe 5 , 

Dirk Grimm 3 , Holger Willenbring 1,6 ; 1 Institute of Regeneration 

Medicine, University of California San Francisco, San Francisco, 

CA; 2 Department of Pediatrics, Division of Gastroenterology, 

Hepatology, and Nutrition, University of California San Francisco, 

San Francisco, CA; 3 Department of Infectious Diseases, Cluster of 

Excellence CellNetworks, BioQuant BQ0030, Heidelberg University 

Hospital, Heidelberg, Germany; 4 4Department of Surgery, 

Division of Pediatric Surgery, University of California San Francisco, 

San Francisco, CA; 5 Department of Medicine,, Columbia 

University, New York, NC; 6 Department of Surgery, Division of 

Transplantation, University of California San Francisco, San Francisco, 

CA 

Repeated hepatocyte loss in chronic liver injury can exceed 

the regenerative capabilities of hepatocytes and lead to liver 

fibrosis, a form of scarring characterized by replacement of 

hepatocytes by collagen produced by myofibroblasts (MFs). 

The structural and molecular changes associated with liver 

fibrosis further impair liver function, leading to liver failure. 

The only cure for advanced liver fibrosis is liver transplantation, 

but donor organs are scarce. As a strategy to replenish 

the hepatocyte mass and limit collagen deposition in the 

chronically injured liver, we developed in vivo reprogramming 

of MFs into hepatocytes by overexpression of hepatic transcription 

factors. To facilitate clinical translation, we delivered 

these genes to MFs using adeno-associated viral (AAV) vectors, 

which are not toxic and do not integrate into the genome. We 

first investigated the feasibility of generating induced hepatocytes 

from primary MFs (MF-iHeps) using AAV vectors expressing 

FOXA1, FOXA2, FOXA3, GATA4, HNF1α and HNF4α in 

vitro. We found that AAV vectors were effective in generating 

expandable MF-iHeps resembling previously reported iHeps 

generated from fibroblasts with integrating retroviral or lentiviral 

vectors. Specifically, MF-iHeps lost most of their original 

cell identity and acquired hepatocyte gene and protein expression 

and functions like low-density lipoprotein uptake, glycogen 

storage and cytochrome P450 activity. Next, we established 

hepatic reprogramming of MFs in vivo. For this we used a 

lineage-tracing mouse model in which MFs and their progeny 

are constitutively labeled. To induce liver fibrosis, we treated 

these mice with carbon tetrachloride. After intravenous injection 

of the AAV vectors we observed the formation of MF-iHeps 

and reduced liver fibrosis. Like primary hepatocytes MF-iHeps 

proliferated in response to liver injury in both CCl4-treated 

and fumarylacetoacetate hydrolase-deficient mice. To assess 

hepatocyte differentiation of MF-iHeps, we isolated them by 

laser-capture microscopy and analyzed their gene expression 

with microarrays. MF-iHeps closely resembled primary hepatocytes, 

with the exception of minimal residual MF identity. We 

confirmed these results using functional assays, including analysis 

of hepatic glucose metabolism, and ascertained stability 

of MF-iHep differentiation in mice followed for 6 months. Our 

results establish repurposing of MFs as a potential new therapy 

for liver fibrosis that not only reduces fibrosis but also increases 

the functional hepatocyte mass. By using AAV vectors, which 

proved effective and safe in liver-directed human gene therapy, 

our strategy lends itself well to clinical translation. 

Disclosures: 

Robert F. Schwabe - Consulting: Merck 

The following authors have nothing to disclose: Milad Rezvani, Regina Español- 

Suñer, Yann Malato, Laure Dumont, Andrew A. Grimm, Eike Kienle, Julia Bindmann, 

Ellen Wiedtke, Syed J. Naqvi, S. C. Derderian, Dirk Grimm, Holger 

Willenbring 

99 

Protective role of miR-122 against acetaminophen toxicity 

is due to suppression of Cyp2e1 and Cyp1a2 

Vivek K. Chowdhary 3 , Huban Kutay 3 , Laura James 2 , William M. 

Lee 1 , Kalpana Ghoshal 3 ; 1 UT Southwestern Medical Center, Dallas, 

TX; 2 University of Arkansas for Medical Science, Little Rock, 

AR; 3 Pathology, Comprehensive Cancer Center, The Ohio State 

University College of Medicine, Columbus, OH 

Approximately 2000 cases of acute liver failure occur annually 

in the United States and acetaminophen (APAP) accounts for 

nearly 50% of cases. miR-122 is the most abundant, conserved 

liver-specific microRNA that maintains metabolic homeostasis 

and functions as a tumor suppressor. Although circulating 

miR-122 is a sensitive biomarker of APAP toxicity in humans 

and rodents, its role in this injury has not been elucidated. To 

investigate whether miR-122 has any protective role against 

APAP toxicity, we gave APAP (500mg/Kg) intraperitoneally 

to the Mir122 fl/fl (WT) and liver-specific miR-122 knockout 

(Mir122 fl/fl ; Alb-Cre) (aka LKO) mice generated in our lab 

(PMID: 22820288), and monitored their survival. Mortality 

rate of LKO mice was significantly higher than that of WT mice 

(P


100 

CX3CR1 is a gatekeeper for intestinal barrier integrity: 

Limiting steatohepatitis by maintaining intestinal 

homeostasis 

Kai M. Schneider 1 , Veerle Bieghs 1 , Felix Heymann 1 , Wei Hu 1 , 

Daniela Dreymueller 2 , Lijun Liao 1 , Mick Frissen 1 , Andreas Ludwig 2 , 

Nikolaus Gassler 3 , Oliver Pabst 4 , Eicke Latz 5,6 , Gernot Sellge 1 , 

John Penders 7 , Frank Tacke 1 , Christian Trautwein 1 ; 1 Department 

of Internal Medicine III, RWTH Aachen University, Aachen, Germany; 

2 Institute of Pharmacology and Toxicology, Medical Faculty, 

University Hospital RWTH Aachen, Aachen, Germany; 3 Institute 

of Pathology, University Hospital RWTH Aachen, Aachen, Germany; 

4 Institute of Molecular Medicine, University Hospital RWTH 

Aachen, Aachen, Germany; 5 Institute of Innate Immunity, University 

Hospital, University of Bonn, Bonn, Germany; 6 Department of 

Medicine, University of Massachusetts Medical School, Worcester, 

MA; 7 Department of Medical Microbiology, School of Nutrition 

and Translational Research in Metabolism, Maastricht University 

Medical Center, Maastricht, Netherlands 

Background: Non-alcoholic fatty liver disease (NAFLD) represents 

the most common liver disease in Western societies 

and is regarded as the hepatic manifestation of the metabolic 

syndrome. The G-Protein-coupled chemokine receptor Cx3cr1 

has been shown to play a central role in many metabolic diseases 

including type-2 diabetes, atherosclerosis and obesity. 

However, the role of Cx3cr1 for NAFLD progression is unclear. 

Recent data indicate that intestinal dysbiosis drives NASH 

development and Cx3cr1 is essential for intestinal homeostasis. 

Based on these findings, we hypothesized that Cx3cr1 

plays a role in regulating the gut-liver axis and therefore has 

implications for NASH progression. Methods: Male wildtype 

(WT) and Cx3cr1 -/- mice were fed either a normal chow diet 

(NCD), high-fat diet (HFD) or methionine-choline-deficient diet 

(MCD) to induce steatohepatitis. For eradicating the intestinal 

microbiota, the drinking water of the mice was supplemented 

with broad-spectrum non-resorbable antibiotics. Cx3cr1-signaling 

was studied using Bone-marrow-derived macrophages 

(BMDMs). Results: On HFD or MCD Cx3cr1 -/- mice showed 

more severe hepatic steatosis and inflammation, as well as systemic 

glucose intolerance compared to WT controls. Mechanistically, 

in-vitro experiments with BMDMs identified Cx3cr1 as 

a regulator of macrophage homeostasis by modulating inflammasome 

activation. Accordingly, Cx3cr1 deficiency in mice 

was associated with significantly altered intestinal microbiota 

composition, which was linked to an impaired intestinal barrier 

including thinner colonic mucus layers, reduction of tight junction 

expression and a decrease in colonic resident phagocytic 

macrophages. Concomitantly, these intestinal changes led to 

an increased translocation of endotoxin and stronger activation 

of pathogen recognition receptors (PRRs) in livers of Cx3cr1 -/- 

mice, thereby triggering an enhanced inflammatory response 

in the liver. Strikingly, depletion of the intestinal microbiota 

by administration of broad-spectrum antibiotics (AB) did not 

only suppress the number of infiltrating macrophages, but also 

promoted a restorative phenotype of liver macrophages. Consequently, 

AB-treated mice demonstrated a marked improvement 

of steatohepatitis and glucose tolerance. Conclusion: Our 

data demonstrate that microbiota-mediated activation of the 

innate immune responses via Cx3cr1 is crucial for controlling 

steatohepatitis progression, thereby recognizing CX3CR1 as 

an essential gatekeeper in this scenario. 

Disclosures: 

Frank Tacke - Advisory Committees or Review Panels: Tobira; Grant/Research 

Support: Novartis, Noxxon; Speaking and Teaching: BMS, Gilead, Falk, MSD, 

Janssen, Abbvie 

Christian Trautwein - Grant/Research Support: BMS, Novartis, BMS, Novartis; 

Speaking and Teaching: Roche, BMS, Roche, BMS 

The following authors have nothing to disclose: Kai M. Schneider, Veerle Bieghs, 

Felix Heymann, Wei Hu, Daniela Dreymueller, Lijun Liao, Mick Frissen, Andreas 

Ludwig, Nikolaus Gassler, Oliver Pabst, Eicke Latz, Gernot Sellge, John Penders 

101 

Clonal analysis of the human liver reveals adaptable 

stem cell dynamics 

Malcolm Alison, Biancastella Cereser, Hemant Kocher, Stuart A. 

McDonald; Centre for Tumour Biology, Barts Cancer Institute, London, 

United Kingdom 

The study of cell lineages through heritable genetic lineage 

tracing is well established in experimental animals. However, 

using such techniques it is still unclear whether the liver conforms 

to a stem cell and lineage system, as seen for example 

in the bone marrow and gut. We have sought to resolve 

this question in the human liver using a variety of investigative 

techniques. The mitochondrial genome is highly prone 

to non-pathogenic mutations resulting in a deficiency in cytochrome 

c oxidase (CCO). Such mutations can be used as markers 

of clonal expansion and understanding the phylogenetics 

of these mutations will help in identifying stem cell derived 

clonal populations and their dynamics. Analysis of many hundreds 

of periportal and hepatic venous regions revealed that 

the majority of CCO-deficient cell patches abutted portal tracts, 

and when adjacent to hepatic veins the CCO-deficient patch 

typically extended from the portal tract. Furthermore, CCO-deficient 

patches were associated with trifurcating terminal portal 

tracts, but not with conducting portal tracts. Secondly we analyzed 

the methylation patterns of CpG islands in the promoter 

regions of non-expressed genes in CCO-deficient cell patches 

with respect to the portal tract-hepatic vein axis. Patterns from 

microdissected areas within CCO-deficient patches became 

diverse a short distance away from portal tracts, suggesting 

that most periportal and centrilobular hepatocytes are of similar 

mitotic age and only hepatocytes in the immediate periportal 

zone revealed patterns that were proportional to the size 

of the clone. Next generation sequencing (NGS) of human 

mtDNA was also employed to determine the clonal dynamics in 

CCO-deficient patches. Some mutations were common (clonal) 

to all cells within a patch, while some were confined (private) 

to cells at different points through the patch. Overall the methylation 

and NGS data suggest that CCO patches developed 

initially as a rapid expansion (perhaps through acute damage) 

which then ceased and allowed cell by cell diversity to accrue. 

Interestingly, the centrilobular regions revealed a higher mutation 

burden, indicative of these being older cells having had 

more time for subclonal expansion of mutations. Our studies 

employing CCO activity, promoter methylation and NGS are 

consistent with the human liver being a lineage system, presumably 

emanating from periportal hepatic progenitor cells, 

the so-called streaming liver. However, this process appears to 

be very slow in normal homeostasis and rapid perhaps during 

periods of acute damage and suggests an adaptable stem cell 

organisation. 

Disclosures: 

The following authors have nothing to disclose: Malcolm Alison, Biancastella 

Cereser, Hemant Kocher, Stuart A. McDonald


102 

MicroRNA-21 is a Potential Link between Non-alcoholic 

Fatty Liver Disease and Hepatocellular Carcinoma via 

Modulation of the HBP1-p53-Srebp1c Pathway 

Heng Wu, Guisheng Song; Medicine, University of Minnesota, 

Minneapolis, MN 

Background: Nonalcoholic fatty liver disease (NAFLD) is a 

major risk factor for hepatocellular carcinoma (HCC). However, 

the mechanistic pathways that link both disorders are 

essentially unknown. Our study was designed to investigate 

the role of microRNA-21 in the pathogenesis of NAFLD and its 

potential involvement in HCC. Methods: Wild-type mice maintained 

on a high fat diet (HFD) received tail-vein injections of 

microRNA-21-ASO (anti-sense oligonucleotide) or miR-21 mismatched 

ASO for 4 or 8 weeks. Livers were collected after that 

time period for lipid content and gene expression analysis. The 

role of microRNA-21 in carcinogenesis was analyzed by softagar 

colony formation, cell cycle analysis, and xenograft tumor 

assay. Results: The expression of microRNA-21 was increased 

in the livers of HFD-treated mice and human HepG2 cells incubated 

with fatty acid. MicroRNA-21 knockdown in those mice 

and HepG2 cells impaired lipid accumulation and growth of 

xenograft tumor. Further studies revealed that Hbp1 was a 

novel target of microRNA-21 and a transcriptional activator of 

p53. It is well-established that p53 is a tumor suppressor and 

an inhibitor of lipogenesis by inhibiting Srebp1c. As expected, 

microRNA-21 knockdown led to increased HBP1 and p53 and 

subsequently reduced lipogenesis and delayed G1/S transition, 

and the additional treatment of HBP1-siRNA antagonized 

the effect of microRNA-21-ASO, suggesting that HBP1 

mediated the inhibitory effects of microRNA-21-ASO on both 

hepatic lipid accumulation and hepatocarcinogenesis. Mechanistically, 

microRNA-21 knockdown induced p53 transcription, 

which subsequently reduced expression of genes controlling 

lipogenesis and cell cycle transition. In contrast, the opposite 

result was observed with overexpression of microRNA-21, 

which prevented p53 transcription. Conclusion: Our findings 

reveal a novel mechanism by which microRNA-21, in part, 

promotes hepatic lipid accumulation and cancer progression 

by interacting with the Hbp1-p53-Srebp1c pathway, and suggest 

the potential therapeutic value of microRNA-21-ASO for 

both disorders. 

Disclosures: 

The following authors have nothing to disclose: Heng Wu, Guisheng Song 

103 

Comparison And Outcomes Of 5% Albumin Vs 0.9% 

Normal Saline Fluid Resuscitation In Cirrhotics Presenting 

With Sepsis Induced Hypotension – A Randomized 

Controlled Trial - Fluid Resuscitation In Septic Shock In 

Cirrhosis (FRISC Protocol) 

Cyriac A. Philips 1 , Ashok K. Choudhury 1 , Amrish Sahney 1 , Rakhi 

Maiwall 1 , Lalita G. Mitra 2 , Shiv K. Sarin 1 ; 1 Hepatology and Transplant 

Medicine, Institute of Liver and Biliary Sciences, New Delhi, 

India; 2 Anesthesia and Critical Care, Institute of Liver and Biliary 

Sciences, New Delhi, India 

Background&Aims:Fluid resuscitation and choice of fluid for 

use in sepsis induced hypotension (SIH)is generally done as per 

Surviving Sepsis Guidelines (SSG). There is however, no data 

on fluid resuscitation protocols in cirrhosis patients. We investigated 

the efficacy, hemodynamic and perfusion effects of 

human albumin (HA) versus 0.9% normal saline(NS)in cirrhotics 

presenting with SIH. Patients&Methods: 308 patients of cirrhosis 

with SIH were randomized to receive either HA(5%,250ml 

bolus over 15 mins,n=154) or NS(30mL/kg over 30 mins);primary 

end-point (PE) being increase in mean arterial pressure 

(MAP)>65 mm Hg at 3 hr; secondary end-points of effects 

on heart rate(HR),lactate [delta lactate (dLAC), lactate clearance 

(cLAC)], urine output (UO) from baseline, at 1, 2 and 

3 hr and survival at 1 wk. Results:154 patients each, in HA 

(males,76%,mean age 49.7 yr) or NS group (79.2%,47 yr);(- 

MAP-53.8±6.2/54.4±6 mmHg,HR–93.6±22.6/93.4±16.8/ 

bpm,LAC–6.94±2.2/6.83±2.6 mmol/L) and severity scores 

(CTP11.8±1.9/12.2±1.9,MELD 32±8.6/30±8.3,SOFA- 

9.99±2.5/10.4±2.8) had comparable baseline parameters. 

MAP>65mmHg at 1hr and sustenance of MAP at 3hr were in 

higher(p


idly. Our objective was to project the number of HCV patients 

needing treatment in 2015 and beyond, and the long-term 

health outcomes under different treatment penetration rates. 

Method: We used our previously published Hepatitis C Disease 

Burden Simulation model (HEP-SIM), which projected the 

changing prevalence of HCV in the United States. The HEP-SIM 

model was validated with NHANES studies and other published 

data. We simulated the current clinical management of 

HCV from 2001 onwards, which included risk-based screening 

until 2013 and addition of birth-cohort screening afterwards. 

We modeled antiviral treatment in different waves starting with 

peginterferon-ribavirin (PEG-RBV) until 2011, followed by the 

launch of boceprevir/telaprevir in 2012, sofosbuvir/simeprevir 

in 2014, and finally oral DAAs in 2015. We also implemented 

changes in insurance status because of the Affordable Care 

Act. We projected the number of patients needing treatment 

in 2015 and beyond under varying treatment capacity/penetration 

scenarios. We also projected patients’ fibrosis score, 

HCV awareness status, and access to insurance. Results: We 

estimated that in 2015, 2 million noninstitutionalized patients 

would be chronically infected and viremic. Among these 

patients, 1.1 million would be potential DAA candidates, i.e., 

aware of their HCV status and insured. At the current annual 

treatment capacity of 200,000 patients, it will take at least 10 

years to treat the majority of the patients. By 2025, the number 

of treatment candidates would decline to fewer than 50,000 

patients. Even in the DAA era, 320,000 patients would die 

because of HCV, 32,000 patients would get liver transplants, 

202,000 would develop decompensated cirrhosis (DC) and 

156,000 would progress to hepatocellular carcinoma (HCC) 

by 2050. Doubling the annual treatment capacity could avoid 

8,000 deaths, 700 transplants, 7,000 DC and 4,000 HCC 

cases by 2050. Conclusions: Though the majority of patients 

aware of their HCV will be treated in the next 10 years, the 

HCV burden would still remain substantial unless aggressive 

screening and treatment policies are implemented. Increasing 

HCV treatment capacity is essential to decrease disease burden 

and improve health outcomes of HCV patients in the US, and 

decrease health resource utilization. 

Disclosures: 

Jagpreet Chhatwal - Consulting: Merck & Co., Inc., Gilead, Complete HEOR 

Solutions; Grant/Research Support: NIH/National Center for Advancing Translational 

Sciences 

The following authors have nothing to disclose: Xiaojie Wang, Fasiha Kanwal, 

Mina Kabiri, Turgay Ayer, Julie M. Donohue, Mark S. Roberts 

105 

An international, phase 2 randomized controlled trial of 

the dual PPAR α-δ agonist GFT505 in adult patients with 

NASH 

Vlad Ratziu 1 , Stephen A. Harrison 2 , Sven M. Francque 3 , Pierre 

Bedossa 4 , Lawrence Serfaty 5 , Manuel Romero-Gomez 6 , Paul 

Cales 7 , Manal F. Abdelmalek 8 , Stephen H. Caldwell 9 , Joost 

Drenth 10 , Quentin M. Anstee 11 , Dean W. Hum 12 , Rémy Hanf 12 , 

Alice Roudot 12 , Sophie Megnien 12 , Bart Staels 13 , Arun J. Sanyal 14 ; 

1 Hepatology, Hopital Pitie Salpetriere, Paris, France; 2 Department 

of Medicine, Gastroenterology & Hepatology Service,, Brooke 

Army Medical Center, Fort Sam Houston, TX; 3 Department of Gastroenterology 

& Hepatology, Antwerp University Hospital, University 

of Antwerp, Antwerp, Belgium; 4 Department of Pathology, 

Hôpital Beaujon, University Paris-Denis Diderot, Clichy, France; 

5 Service d’Hépatologie,, Hôpital Saint-Antoine, APHP, UPMC Paris 

6, Paris, France; 6 Hospital Universitario de Valme, Unit for the 

Clinical Management of Digestive Diseases and CIBERehd, Sevilla, 

Spain; 7 Hepatology Department, University Hospital & LUNAM 

University, Angers, France., Angers, France; 8 Duke University, 

Duke university, Durham, NC; 9 University of Virginia, Gastroenterology 

& hepatology Division, Charlottesville, VA; 10 Radboud 

University Medical Center, Department of Gastroenterology and 

Hepatology, Nijmegen, Netherlands; 11 Institute of Cellular Medicine,, 

Faculty of Medical Sciences, Newcastle University, Newcastle 

upon Tyne, United Kingdom; 12 Genfit SA, Loos, France; 

13 Université Lille 2, INSERM U1011, European Genomic Institute 

for Diabetes (EGID), Institut Pasteur de Lille, Lille, France, Lille, 

France; 14 Virginia Commonwealth University, Richmond, VA 

Peroxisome proliferator-activated receptor α-δ dual agonists, 

such as GFT505, are a promising therapy for NASH as they 

improve hepatic insulin sensitivity, glucose homeostasis, lipid 

metabolism, and inflammation. Methods. In this randomized 

controlled trial (56 European and US centers) 274 patients (pts) 

(full analysis set, FAS) with histologically-defined non-cirrhotic 

NASH received GFT505 80 mg or 120 mg QD vs placebo (PLB) 

for one year. The primary outcome was resolution of NASH 

without worsening of fibrosis. Data were analyzed according 

to baseline severity (histological NAS score) and center effect. 

Biopsies were read by a single pathologist. Results. 237 pts 

had entry and end-of-treatment biopsies (ITT population). While 

the a priori primary endpoint did not meet significance, after 

controlling for baseline severity and center effect, pts in the 

120 mg arm had a 1.94 (CI 1.08-3.48, p=0.027) higher relative 

risk (RR) of achieving the primary end-point compared to 

PLB, while the RR was 1.68 (0.92-3.05, p=0.091) for the 80 

mg arm. Results were similar in the FAS where pts missing the 

second biopsy were counted as failures. In pts with moderate 

activity (NAS 4 or 5) the response rate was 27.5% in the 120 

mg arm vs. 19.5% for the PLB arm. In those with severe activity 

(NAS>5) it was 14.8% vs. 0%, respectively. In the 120 pts 

with NAS>4 from centers that recruited >1 patient/arm, the 

response rate was 29% and 5% in the 120 mg and PLB arms, 

respectively, p=0.01. A >2 point NAS reduction was obtained 

in 48% and 21% of patients respectively, p=0.01. Compared 

to PLB, the 120 mg arm improved ballooning (45% vs. 23%, 

p=0.02), inflammation (55% vs. 33%, p=0.05) and steatosis 

(35.5% vs. 18%, NS). In the 120 mg arm, resolution of NASH, 

resulted in a significant improvement in fibrosis (mean change 

-0.67 vs. +0.09 in non-responders, p


as HbA1c and FFA in diabetic pts, all on top of standard of 

care therapies. Tolerability was excellent without weight gain, 

cardiac events or safety signal. Conclusion. In NASH patients 

120 mg daily of GFT505 induced histological improvement 

and resolution of NASH, significantly more often than PLB. 

The excellent safety and tolerability and the improvement in 

cardiometabolic risk profile makes GFT505 an ideal drug candidate 

to be tested in phase 3 trials 

Disclosures: 

Vlad Ratziu - Advisory Committees or Review Panels: GalMed, Abbott, Genfit, 

Enterome, Gilead; Consulting: Tobira, Intercept, Exalenz, Sanofi-Synthelabo, 

Boehringer-Ingelheim 

Stephen A. Harrison - Advisory Committees or Review Panels: Merck, Nimbus 

Discovery, Fibrogen, RuiYi, CLDF; Consulting: NGM Biopharmaceuticals; Speaking 

and Teaching: Gilead, Abbvie, Janssen, CLDF 

Lawrence Serfaty - Board Membership: BMS, Gilead; Consulting: Merck; Speaking 

and Teaching: Roche, Janssen, Merck, Janssen, BMS, Gilead 

Manuel Romero-Gomez - Advisory Committees or Review Panels: Roche Farma,SA., 

MSD, S.A., Janssen, S.A., Abbott, S.A.; Grant/Research Support: Ferrer, 

S.A. 

Paul Cales - Consulting: BioLiveScale 

Manal F. Abdelmalek - Consulting: Islet Sciences; Grant/Research Support: 

Tobira, Gilead Sciences, NIH/NIDDK, Synageva, Genfit Pharmaceuticals, 

Immuron, Galmed, TaiwanJ Pharma, Intercept, NGM Pharmaceuticals 



NGM 

Quentin M. Anstee - Advisory Committees or Review Panels: Genfit, Intercept, 

Raptor; Grant/Research Support: GSK; Speaking and Teaching: Abbott Laboratories 

Dean W. Hum - Management Position: Genfit 

Rémy Hanf - Management Position: GENFIT 

Alice Roudot - Employment: GENFIT 

Sophie Megnien - Employment: GENFIT 

Bart Staels - Advisory Committees or Review Panels: MSD; Consulting: Genfit 

Arun J. Sanyal - Advisory Committees or Review Panels: Bristol Myers, Gilead, 

Genfit, Abbott, Ikaria, Exhalenz; Consulting: Salix, Immuron, Exhalenz, Nimbus, 

Genentech, Echosens, Takeda, Merck, Enanta, Zafgen, JD Pharma, Islet 

Sciences; Grant/Research Support: Salix, Genentech, Intercept, Ikaria, Takeda, 

GalMed, Novartis, Gilead, Tobira; Independent Contractor: UpToDate, Elsevier 

The following authors have nothing to disclose: Sven M. Francque, Pierre 

Bedossa, Joost Drenth 

106 

NGM282, A Novel Variant of FGF-19, Demonstrates 

Biologic Activity in Primary Biliary Cirrhosis Patients 

with an Incomplete Response to Ursodeoxycholic Acid: 

Results of a Phase 2 Multicenter, Randomized, Double 

Blinded, Placebo Controlled Trial 

Marlyn J. Mayo 3 , Alan J. Wigg 4 , Stuart K. Roberts 5 , Hays 

Arnold 19 , Tarek I. Hassanein 7 , Barbara A. Leggett 8 , John P. Bate 9 , 

Martin Weltman 10 , Elizabeth J. Carey 11 , Andrew J. Muir 12 , Geoff 

McCaughan 14 , Steven J. Bollipo 15 , Stuart C. Gordon 16 , Peter W. 

Angus 13 , Stephen Riordan 17 , Mitchell L. Shiffman 18 , Elisa Young 6 , 

Lei Ling 1 , Jian Luo 1 , Michael Elliott 1 , Stephen Rossi 1 , Alex M. 

DePaoli 1 , Alex J. Thompson 2 ; 1 NGM Biopharmaceuticals, Inc., 

South San Francisco, CA; 2 St. Vincent’s Hospital, Melbourne, VIC, 

Australia; 3 UT Southwestern, Dallas, TX; 4 Flinders Medical Center, 

Adelaide, SA, Australia; 5 Alfred Hospital, Melbourne, VIC, 

Australia; 6 Novotech, Sydney, NSW, Australia; 7 Southern California 

Liver Research Center, Coronado, CA; 8 Royal Brisbane and 

Women’s Hospital, Brisbane, QLD, Australia; 9 Royal Adelaide 

Hospital, Adelaide, SA, Australia; 10 Nepean Hospital, Sydney, 

NSW, Australia; 11 Mayo Clinic - Arizona, Scottsdale, AZ; 12 Duke 

University, Durham, NC; 13 Austin Hospital, Heidelberg, VIC, Australia; 

14 University of Sydney, Sydney, NSW, Australia; 15 John 

Hunter Hospital, New Lambton, NSW, Australia; 16 Henry Ford 

Hospital, Detroit, MI; 17 Prince of Wales Hospital, Sydney, NSW, 

Australia; 18 Liver Institute of Virginia, Richmond, VA; 19 Digestive 

Research Center, San Antonio, TX 

Background: PBC patients with an incomplete biochemical 

response to ursodeoxycholic acid (UDCA) are at increased 

risk for disease progression and with limited treatment options. 

NGM282 is a novel engineered variant of FGF-19 that inhibits 

the CYP7A1-mediated bile acid (BA) synthesis in animals and 

healthy volunteers. The biologic activity was therefore evaluated 

in PBC patients. Methods: 45 subjects with a baseline 

(BL) alkaline phosphatase (ALP) >1.67xULN after 1yr of UDCA 

were randomized to NGM282 0.3 or 3mg vs placebo (PBO) 

as a daily SC injection for 28d. Change from BL ALP was the 

primary endpoint, with key secondary efficacy endpoints of 

liver chemistries and BA synthesis (7a-hydroxy-4-cholesten-3- 

one or C4). Pruritus was measured at all study visits with the 

5D Itch and Visual Analogue Score (VAS). Post-hoc sub-analyses 

evaluated ALP response by BL ALP< or >3xULN. Results: 

All study arms were balanced for typical PBC patient characteristics 

(female=91%, mean age=56y, mean BL ALP=297 

IU/L, mean UDCA dose=15mg/kg/d). Pruritus at BL was 54%, 

64% and 67% of PBO, 0.3mg and 3mg arms, respectively. 

NGM282-treated subjects had a significant, dose-dependent 

reduction in ALP vs PBO (p3xULN (Table 1). Significant reductions were also 

seen in liver chemistries (p90% supporting a potent biologic effect. NGM282 

was safe and well tolerated with no observed safety signals. 

Adverse events occurring >10% of both NGM282 treatment 

arms were diarrhea (PBO=6.7%, 0.3mg=21.4%, 3mg=25%), 

headache (PBO=6.7%, 0.3mg=14.3%, 3mg=25%) and nausea 

(PBO=6.7%, 0.3mg=14.3%, 3 mg=12.5%), the majority 

of which were mild. There was no clinically significant evidence 

of drug-induced pruritus by either VAS or 5D itch. Conclusions: 

These data demonstrate the biologic activity of NGM282 in 

PBC and support a therapeutic potential in other BA-mediated 

diseases. Studies are ongoing of longer duration, increased 

dose and BL predictors of response to identify optimal PBC 

patient populations for potential treatment with NGM282.


Table 1 Mean change in liver and BA Parameters 

Disclosures: 

Marlyn J. Mayo - Grant/Research Support: Intercept, Salix, NGM, Lumena, 

Gilead 

Stuart K. Roberts - Board Membership: AbbVie, Gilead 








Barbara A. Leggett - Advisory Committees or Review Panels: MSD, MSD, MSD, 

MSD; Speaking and Teaching: Roche, Roche, Roche, Roche, Gilead 




Geoff McCaughan - Advisory Committees or Review Panels: Gilead 

Stuart C. Gordon - Advisory Committees or Review Panels: Janssen; Consulting: 

Merck, Gilead, BMS, CVS Caremark, Amgen, AbbVie; Grant/Research Support: 

Merck, Gilead, AbbVie, Intercept Pharmaceuticals, Exalenz Sciences, Inc., BMS 

Peter W. Angus - Advisory Committees or Review Panels: Gilead Sciences, BMS; 

Grant/Research Support: Gilead sciences 

Mitchell L. Shiffman - Advisory Committees or Review Panels: Merck, Gilead, 

Boehringer-Ingelheim, Bristol-Myers-Squibb, Abbvie, Janssen, Acchillion; Consulting: 

Roche/Genentech; Grant/Research Support: Merck, Gilead, Boehringer-Ingelheim, 

Bristol-Myers-Squibb, Abbvie, Beckman-Coulter, Achillion, Lumena, 

Intercept, Novartis, Gen-Probe; Speaking and Teaching: Roche/Genentech, 

Merck, Gilead, Abbvie, Janssen, Bayer 

Lei Ling - Employment: NGM Biopharmaceuticals, Inc. 

Jian Luo - Employment: NGM Biopharmaceuticals 

Michael Elliott - Employment: NGM; Stock Shareholder: NGM 

Stephen Rossi - Employment: NGM Biopharmaceuticals, Inc; Stock Shareholder: 

NGM Biopharmaceuticals, Gilead Sciences 

Alex M. DePaoli - Employment: NGM Biopharmaceuticals 

Alex J. Thompson - Advisory Committees or Review Panels: Gilead, Abbvie, 

BMS, Merck, Spring Bank Pharmaceuticals, Arrowhead, Roche; Grant/Research 

Support: Gilead, Abbvie, BMS, Merck; Speaking and Teaching: Roche, Gilead, 

Abbvie, BMS 

The following authors have nothing to disclose: Alan J. Wigg, Hays Arnold, John 

P. Bate, Martin Weltman, Elizabeth J. Carey, Steven J. Bollipo, Stephen Riordan, 

Elisa Young 

in non-diabetic NASH derived from the PIVENS vitamin E and 

placebo groups and from non-diabetics in the FLINT placebo 

group. Methods: Two efficacy measures from FLINT were 

applied to our pooled data: histologic improvement, defined 

as ≥ 2 point improvement in NAS with no worsening of fibrosis 

or NASH resolution. Safety estimates paralleled those used in 

FLINT and included incidence of cardiac events and changes in 

lipid levels. Logistic regression models were used to summarize 

the odds ratio (OR) effects, confidence limits, and p-values on 

the pooled vitamin E treatment vs no vitamin E treatment efficacy 

estimates; Fisher’s exact test was used to assess cardiac 

events. Results: A total of 250 patients were randomized to 

vitamin E (n=80) or placebo (n=72) in PIVENS or to placebo 

(n=98) in FLINT and had both baseline and end-treatment liver 

biopsies; 53 had diabetes (21%) and 197 were non-diabetic 

(79%); 105 (42%) received vitamin E and 145 (58%) did not 

in the PIVENS or FLINT trials. Vitamin E use was associated 

with histologic improvement in diabetic (OR 4.4, 95% CI 1.1, 

18.0, p=0.04) and non-diabetic patients (OR 3.1, 95% CI 

1.7, 5.8, p


decompensation, reduced risk for hepatocellular carcinoma, 

and reduced liver-related mortality. We propose that cure of 

CHC (defined as SVR of at least 12 weeks) can also lead to 

regression of advanced fibrosis and/or cirrhosis and that Fibroscan 

elastography may be used to confirm reversal of fibrosis. 

METHODS: We conducted a retrospective chart review to 

identify patients treated for CHC with concomitant advanced 

fibrosis or cirrhosis based on clinical features, Fibroscan scores 

and/or biopsy. Advanced fibrosis was defined as FibroScan 

score 11-13.9Kpa and cirrhosis was defined as Fibroscan 

score > 14KPa. This was followed by prospective and retrospective 

Fibroscan and clinical data collection at 6 month intervals, 

up to 14 years. RESULTS: 201 subjects were enrolled, 58 

are currently eligible for analysis with at least 2 measurements 

of fibrosis. Mean age was 60.7 years, 70.7% were male, and 

most were Non-Hispanic (81%) white (91.4%). At baseline 

25.9% had hypertension, 12.1 % had diabetes, 12.1% had 

hyperlipidemia and the average BMI was 27.7 (SD 5). Of 

the 34 subjects who had cirrhosis at baseline, 18 (52.9%) 

demonstrated improvement by Fibroscan, with a median time 

to improvement of 2.8 years (IQR 1.0-3.5). Improvement was 

defined as a change in stage of fibrosis rather than a change in 

Fibroscan score. Of the 17 subjects who had advanced fibrosis, 

16 (66.7%) demonstrated improvement, with a median time of 

2.0 years (IQR 1.2-2.5). Gender, BMI, age, individual medical 

problems and HCV genotype were not found to be associated 

with improvement or worsening of baseline liver disease. ALT, 

albumin and INR changes did not correlate with fibrosis regression. 

However, platelet count change (increased counts in subjects 

who demonstrated improved liver disease) approached 

statistical significance with a p=0.06 in subjects with cirrhosis. 

CONCLUSION: The majority of our subjects, 34/58 (58.6%), 

with advanced fibrosis or cirrhosis demonstrated improvement 

by Fibroscan scores after a median follow-up (after treatment 

of CHC) of 2.5 years (range: 0.5-14 years, IQR: 1- 3.5 years). 

We have yet been not able to establish factors that may predict 

improvement of cirrhosis or advanced fibrosis. Most enrolled 

patients are still awaiting follow-up Fibroscan test results anticipated 

to be completed in 2015. 

Disclosures: 

Catherine T. Frenette - Speaking and Teaching: Bayer, Salix, Gilead; Stock 


Paul J. Pockros - Advisory Committees or Review Panels: Janssen, Merck, BMS, 

Gilead, AbbVie; Consulting: Lumena, Beckman Coulter; Grant/Research Support: 

Intercept, Janssen, BMS, Gilead, Lumena, Beckman Coulter, AbbVie, RMS, 

Merck; Speaking and Teaching: AbbVie, Janssen, Gilead 

The following authors have nothing to disclose: Ana Maria Crissien, William B. 

Minteer, Jason J. Pan 

109 

Circulating extracellular vesicles and their microRNA 

cargos are potential novel biomarkers with a functional 

role of miR-122 in monocyte activation in alcoholic hepatitis 

Banishree Saha, Fatemah Momen-Heravi, Shashi Bala, Karen 

Kodys, Gyongyi Szabo; Medicine, UMASS Med School, Worcester, 

MA 

Purpose: Alcohol and its metabolites induce hepatocyte damage 

and recruitment of inflammatory monocytes/macrophages 

and neutrophils leading to alcoholic hepatitis. Currently there 

is no reliable biomarker of alcoholic hepatitis. Extracellular 

vesicles (EVs) found in the circulation carry mRNA, microRNA 

(miRs) and proteins. EVs can serve as biomarkers and mediate 

intercellular communication. miR-122 is abundantly expressed 

in hepatocytes, not in immune cells and increased levels of circulating 

miR-122 were found in liver injury. We hypothesized 

that EV-associated miRs can serve as biomarkers and modulate 

intercellular signaling between hepatocytes and immune 

cells in alcoholic hepatitis. Methods: EVs were isolated from 

chronic alcohol-fed (5 weeks of Lieber DeCarli diet) or pair-fed 

mice sera and from serum of patients with alcoholic hepatitis. 

EVs were characterized by transmission electron microscopy, 

western blot, nanoparticle tracking analysis system and 

miRNA analysis. Results: The total number of circulating EVs 

was significantly increased in alcohol-fed mice as compared 

to control mice. Exosomes (40-150nm) represented most of 

the EVs (~80%). MicroRNA array of circulating EVs revealed 

a significant increase of 7 inflammatory miRs including: miR- 

192, 122, 30a, 744, 1246, 30b and miR-130a in alcohol-fed 

mice compared to controls. The ROC analyses indicated excellent 

diagnostic value of miR-192, 122, and 30a to identify 

alcohol-induced liver injury. In patients with acute alcoholic 

hepatitis, we found a significant increase in the number of 

circulating EVs compared to normal controls and miR-192 and 

miR-30a were significantly increased in the EVs from alcoholic 

hepatitis patients. Serum miR-122 was increased after alcohol 

binge drinking. In the liver, miR-122 is abundantly expressed 

in hepatocytes and monocytes/macrophages have low levels. 

In vitro experiments revealed that exosomes derived from 

ethanol-treated human hepatocytes were taken up by monocytes 

and transferred mature miR-122 into monocytes. This horizontally 

transferred miR-122 inhibited the hemeoxygenase-1 

expression, a target of miR-122 and sensitized monocytes to 

LPS stimulation to increase production of pro-inflammatory cytokines, 

TNF-α and IL-1β; all of these effects were inhibited by 

exosome-mediated delivery of a miR-122 inhibitor in monocytes. 

Conclusion: Elevated levels of EVs and their miR signature 

could serve as biomarkers of alcoholic hepatitis. This study 

reveals a novel EV-mediated mechanism of alcohol-induced 

communication between hepatocytes and monocytes by transferring 

hepatocyte-derived miR-122 that reprograms monocytes 

promoting inflammation in alcoholic hepatitis. 

Disclosures: 

The following authors have nothing to disclose: Banishree Saha, Fatemah 

Momen-Heravi, Shashi Bala, Karen Kodys, Gyongyi Szabo 

110 

Fat-specific Protein 27/CIDEC Promotes Alcoholic Steatohepatitis 

in Mice and Humans 

Ming-Jiang Xu 1 , Yan Cai 1 , Hua Wang 1 , José T. Altamirano 2 , 

Gemma Odena 3 , Frank J. Gonzalez 4 , Ramon Bataller 3 , Bin Gao 1 ; 

1 NIAAA, NIH, Rockville, MD; 2 Vall d’Hebron Institut de Recerca, 

Barcelona, Spain; 3 University of North Carolina, Chapel Hill, NC; 

4 National Institutes of Health, Bethesda, MD 

Objectives: Alcoholic steatohepatitis (ASH) is the progressive 

form of alcoholic liver disease that leads to cirrhosis and 

hepatocellular carcinoma. There is an urgent need to identify 

molecular drivers in order to develop targeted therapies. 

Here the functions of mouse fat-specific protein 27 (Fsp27)/ 

human cell death activator CIDEC (the human homologue of 

Fsp27) were investigated. Methods and results: We developed 

a mouse model with chronic (8 weeks)-plus-binge ethanol feeding, 

which mimics the drinking patterns of alcoholic hepatitis 

(AH) patients, produced severe ASH and mild fibrosis. Microarray 

analyses revealed that mouse Fsp27/human CIDEC gene 

was increased in this animal model and human ASH samples. 

Fsp27 is expressed at high levels in adipose tissues but at 

very low levels in normal liver. Chronic-plus-binge ethanol feeding 

markedly upregulated hepatic Fsp27 mRNA and protein 

expression. Silencing the Fsp27 gene by shRNA or genetic 

disruption ameliorated chronic-plus-binge ethanol-induced


ASH. Inhibition of peroxisome proliferator-activated receptor 

g (PPARg) or cyclic-AMP-responsive-element binding protein H 

(CREBH) attenuated chronic-plus-binge ethanol-induced elevation 

of Fsp27a and FSP27b mRNA, respectively, and subsequently 

ameliorated liver injury. Overexpression of Fsp27 and 

ethanol exposure synergistically induced mitochondrial reactive 

oxygen species production and hepatocyte injury in vivo and 

in vitro, which is likely owing to the mitochondrial location of 

FSP27 leading to the decreased mitochondrial complex I activity. 

Finally, hepatic expression of CIDEC mRNA was elevated 

and positively correlated with hepatic steatosis, disease severity, 

and mortality in AH patients. Conclusion: FSP27/CIDEC 

gene product promotes ASH in chronic-plus-binge ethanol-fed 

mice and in human AH. Targeting CIDEC gene is likely to be a 

novel therapeutic targets for the treatment of ASH. 

overgrowth was similar to WT mice, alcohol-fed Reg3b and 

Reg3g deficient mice showed a significantly higher number of 

mucosa-associated bacteria in the small intestine as compared 

with their respective WT littermates. This was accompanied 

by a significantly increased number of bacteria translocating 

through intestinal epithelial cells and more positive mesenteric 

lymph node cultures in alcohol-fed Reg3b -/- and Reg3g -/- mice. 

Interestingly, absence of Reg3b or Reg3g did not affect the 

intestinal paracellular barrier function as determined by systemic 

LPS level and fecal albumin. Most importantly, Reg3b 

and Reg3g deficient mice showed more alcoholic liver disease 

as assessed by higher plasma ALT levels, increased hepatic 

triglycerides and inflammation. To corroborate our data, we 

generated transgenic mice overexpressing Reg3g under the 

control of the intestine specific villin promoter. Consistent with 

our results, Reg3g transgenic mice showed significantly less 

mucosa-associated bacteria than WT littermates after alcohol 

feeding. Overexpression of intestinal Reg3g suppressed 

translocation of bacteria through intestinal epithelial cells to 

mesenteric lymph nodes, while paracellular permeability and 

systemic LPS levels were not changed after alcohol feeding. 

Reg3g transgenic mice were protected from alcohol-induced 

liver injury, steatosis and inflammation. Conclusion: Intestinal 

Reg3b and Reg3g prevent bacterial colonization of epithelial 

surfaces, which reduces translocation of viable bacteria and 

prevents alcoholic liver disease. Reg3 proteins are novel and 

promising targets in the treatment of alcohol-induced liver disease. 

Disclosures: 

The following authors have nothing to disclose: Lirui Wang, Peng Chen, Lora V. 

Hooper, Bernd Schnabl 

Disclosures: 

Ramon Bataller - Advisory Committees or Review Panels: Sandhill; Consulting: 

VTI, Oncozyme Pharma 

The following authors have nothing to disclose: Ming-Jiang Xu, Yan Cai, Hua 

Wang, José T. Altamirano, Gemma Odena, Frank J. Gonzalez, Bin Gao 

111 

Antimicrobial proteins Reg3b and Reg3g protect mice 

from alcoholic liver disease by preventing bacterial 

translocation 

Lirui Wang 1 , Peng Chen 1 , Lora V. Hooper 2 , Bernd Schnabl 1 ; 

1 University of California San Diego, La Jolla, CA; 2 Department of 

Immunology, The University of Texas Southwestern Medical Center, 

Dallas, TX 

Background: Antimicrobial proteins are secreted by intestinal 

epithelial cells and Paneth cells. They represent the first 

line of defense against pathogens and maintain homeostasis 

with commensal bacteria. We have previously shown that the 

expression of the C-type lectin regenerating islet derived-3 

(Reg3) is suppressed in the small intestine after chronic alcohol 

use in mice and humans, yet functional consequences of 

lower Reg3 expression on the intestinal microbiota, bacterial 

translocation and alcoholic liver disease are unknown. The 

aim of our study was to investigate the role of Reg3b and 

Reg3g in chronic alcoholic liver disease. Methods and Results: 

A Lieber-DeCarli model was used to induce intestinal dysbiosis, 

bacterial translocation and liver disease in mice. After alcohol 

feeding for 8 weeks, wild-type (WT) littermate mice showed 

bacterial overgrowth of the luminal and the mucosa-associated 

microbiota in the small intestine. While luminal bacterial 

112 

Hepatocytes from mice on intragastric feeding model of 

alcoholic steatohepatitis release extracellular vesicles 

with specific microRNA cargo that modulate hepatic 

stellate cell and macrophage phenotype 

Akiko Eguchi 1 , Jihoon Kim 1 , Lucila Ohno-Machado 1 , Hidekazu 

Tsukamoto 2 , Ariel E. Feldstein 1 ; 1 UCSD, La Jolla, CA; 2 USC, Los 

Angeles, CA 

Liver inflammation and fibrosis are key histological features 

associated with prognosis in patients with alcoholic steatohepatitis 

(ASH). Extracellular vesicles (EVs) are released during cell 

stress or demise, can contain a barcode of the cell of origin 

including specific microRNAs and are growingly recognized 

as key cell-to-cell communicators. Here we tested the hypothesis 

that during ASH development, hepatocyte damage release EVs 

with a microRNA signature that can fuse with hepatic stellate 

cells (HSC) and Kupffer cells (KC) to regulate their phenotype. 

Methods: C57/B6 mice were placed on intragastric feeding 

model of continuous ethanol infusion or control diet for 4 weeks 

to reproduce a physiologically relevant model of ASH. The 

extent of steatosis, inflammation, and fibrosis was assessed 

by histological and molecular analyses on liver specimens. 

Isolated hepatocytes (HC) or KC from ASH or control mice 

were incubated in medium-EV free serum and HC- or KC-derived 

EVs were isolated by ultracentrifugation from culture 

medium. A complete characterization of EVs was performed 

by FACS, electron microscopy, dynamic light scattering. HC-EV 

biological function was investigated in isolated primary HSCs 

or KCs derived from C57/B6 mice by cell morphology or 

qPCR. Comprehensive encapsulated miRNA in HC-EVs were 

assessed via transcriptome using illumina miRNA-seq. Results: 

We observed highly significant differences in the levels of HCor 

KC-EVs between control group and ASH (1.3x10 6 HC-EV/


ml of medium in ASH versus 1.4x10 5 HC-EV/ml of medium 

in control, p


ing significant reduction in certain long- (hexadecanoic and 

heptadecanoic), medium (hexanoic and octanoic), and short 

(butanoic) free fatty acids. Remarkably, the levels of octanoic 

acid (known to have antimicrobial properties) were dramatically 

reduced (~48 fold) in mice fed USF+EtOH compared to 

SF+EtOH fed animals. A decline in certain fecal amino acids 

(e.g. serine and glycine) was observed in USF+EtOH fed animals. 

Conclusions: These data support an important role of 

dietary lipids in ALD pathogenesis, and provide insight into 

mechanisms of ALD development. A diet enriched in USF not 

only enhanced alcohol-induced liver injury, but also caused 

major fecal metagenomic and metabolomic changes that may 

play an etiologic role in observed liver injury. Characterization 

of both microbiota composition and function is an important 

approach to investigate host–microbial interaction. Our data 

suggest that dietary lipids can potentially serve as interventions 

for the prevention/treatment of ALD. 

Disclosures: 

Shirish Barve - Speaking and Teaching: Abbott 

Craig J. McClain - Consulting: Vertex, Gilead, Baxter, Celgene, Nestle, Danisco, 

Abbott, Genentech; Grant/Research Support: Ocera, Merck, Glaxo SmithKline; 

Speaking and Teaching: Roche 

The following authors have nothing to disclose: Irina Kirpich, Wenke Feng, Xinmin 

Yin, Xiaoli Wei, Xiang Zhang 

115 

PEGylated TRAIL treatment ameliorates liver cirrhosis in 

rats by targeting activated hepatic stellate cells 

Ogyi Park 1,2 , Yumin Oh 1,2 , Magdalena Swierczewska 1,2 , Jong 

Sung Park 1,2 , Justin Hanes 2 , Martin Pomper 1 , Bin Gao 3 , Seulki 

Lee 1,2 ; 1 The Russell H. Morgan Department of Radiology and 

Radiological Sciences, Johns Hopkins University, Baltimore, MD; 

2 The Center for Nanomedicine at the Wilmer Eye Institute, Johns 

Hopkins University School of Medicine, Baltimore, MD; 3 Laboratory 

of Liver Diseases, National Institute of Alcohol Abuse and 

Alcoholism, National Institutes of Health, Bethesda, MD 

Background: Liver fibrosis is a common outcome of chronic 

liver disease and leads to liver failure and cancer. Still, no targeted 

anti-fibrotic therapy exists at this time. Activated hepatic 

stellate cells (aHSCs) are the originators of liver fibrosis; therefore, 

eradication of aHSCs is a logical strategy to stop and/ 

or reverse liver fibrosis. However, there are no effective strategies 

to specifically deplete apoptosis-resistant aHSCs during 

fibrosis without systemic toxicity. TRAIL is known to selectively 

induce apoptosis in cancer cells or transformed cells by binding 

to its death receptors (DRs) while sparing normal tissue. 

But because recombinant TRAIL suffers from a short half-life 

and low potency, it failed to show efficacy in cancer clinical 

trials. Here we introduce a longer-acting, more stable form 

of TRAIL that shows striking anti-fibrotic activity in rat fibrosis 

and cirrhosis models with no detected hepatotoxicity. Methods: 

We developed PEGylated TRAIL (TRAIL PEG 

) by stabilizing a 

potent homotrimer TRAIL of iLZ-TRAIL with a 5kDa PEG. In in 

vivo studies, SD rats were induced liver fibrosis and cirrhosis 

by CCl 4 

and treated with 4 mg/kg of TRAIL PEG 

daily for 10 

days for fibrosis and two weeks for cirrhosis. In in vitro studies, 

primary human HSCs were culture-activated and TRAIL PEG 

effects were investigated on quiescent and activated HSCs. 

Primary human hepatocytes were used to determine hepatotoxicity 

of TRAIL PEG 

. Results: Intravenously injected TRAIL PEG 

has a markedly extended half-life in non-human primates and 

no toxicity in primary human hepatocytes. We found DRs are 

upregulated in human cirrhotic livers because of aHSCs. In 

vitro primary human aHSCs, but not quiescent HSCs, become 

sensitive to TRAIL PEG 

-induced apoptosis via DR- and DISC-activated 

caspase-8-dependent mechanisms. Intravenous TRAIL PEG 

directly induced apoptosis of aHSCs and ameliorated CCl 4 

-induced 

fibrosis/cirrhosis in rats by simultaneously down-regulating 

multiple key fibrogenic molecules in vivo. TRAIL PEG 

also 

caused selective toxicity to hepatocellular carcinoma (HCC) 

cells. Conclusions: TRAIL-based therapies could serve as firstin-class 

therapeutics for liver fibrosis/cirrhosis, cirrhosis-associated 

HCC and possibly other fibrotic diseases. Our results not 

only introduce the unique activity that TRAIL-based compounds 

have in liver fibrosis but also warrant clinical translation of 

TRAIL PEG 

for severe liver fibrosis therapy. 

Disclosures: 

The following authors have nothing to disclose: Ogyi Park, Yumin Oh, Magdalena 

Swierczewska, Jong Sung Park, Justin Hanes, Martin Pomper, Bin Gao, 

Seulki Lee 

116 

Statins, ACE inhibitors and ARB use is associated with 

reduced mortality and morbidity in chronic liver diseases 

– a nationwide cohort study in Sweden 

Knut Stokkeland 1,2 , Christine Takami Lageborn 3 , Anders Ekbom 4 , 

Jonas Höijer 5 , Matteo Bottai 5 , Per Stål 6 , Karin Söderberg Löfdal 7 ; 

1 Department of Medicine, Visby Hospital, Visby, Sweden; 2 Karolinska 

Institutet, Department of Medicine, Gastroenterology and 

Hepatology, Stockholm, Sweden; 3 Karolinska Institutet, Stockholm, 

Sweden; 4 Department of Medical Epidemiology and Biostatistics, 

Karolinska Institutet, Stockholm, Sweden; 5 Unit of Biostatistics, 

IMM, Karolinska Institutet, Stockholm, Sweden; 6 Division of Hepatology, 

Karolinska Hospital, Stockholm, Sweden; 7 Division of Clinical 

Pharmacology, Department of Laboratory Medicine, Karolinska 

Institutet, Stockholm, Sweden 

Aim: To explore whether current medication in patients with 

chronic liver diseases affect overall mortality, liver-related mortality 

and liver-related morbidity. Methods: We performed a 

register-based cohort study in Sweden with patients with a 

first-time diagnosis of chronic liver disease between 2005 and 

2012. We studied the use of statins, angiotensin converting 

enzyme inhibitors (ACEi), angiotensin receptor blockers (ARB), 

acetylsalicylic acid (ASA), non-steroidal anti-inflammatory 

drugs (NSAID), selective seretonin re-uptake inhibitors (SSRI) 

and antibiotics. We measured total mortality, liver-specific 

mortality and liver-specific morbidity. We used register data 

from the Patient Register, the Prescribed Drug Register and the 

Death Certificate Register. Liver-specific morbidity and mortality 

included all liver diseases, esophageal varices and liver 

cancer. Results: We analyzed 70 546 patients who had been 

seen in hospital out-patient services or hospitalized with chronic 

liver disease for the first time between 2005 and 2012. We 

found a reduction in mortality risk for statin users (n= 8 688) 

with a hazard ratio range between 0.42 (95% CI: 0.28-0.63) 

for patients with autoimmune hepatitis and 0.91 (0.63-1.32) 

for primary biliary cirrhosis with a similar risk reduction for 

liver-specific mortality and liver-related morbidity. There was 

a significant mortality reduction for patients exposed to ARB 

(n= 6204) between 0.62 (0.53-0.72) in alcoholic liver disease 

and 0.91 (0.64-1.32) in primary biliary cirrhosis and a similar 

reduction for patients exposed to ACEi (n=9 714). There was 

a significant reduction of liver morbidity in patients exposed to 

ASA (n= 9 768) for all chronic liver diseases, even among users 

of NSAID (n= 14 119). An increased risk for all-cause mortality 

was seen for almost all liver diseases in patients exposed to 

SSRI (n= 11 048), with a range between 1.25 (1.16-1.35) and 

1.61 (1.30-1.98), and a reduction in risk of liver-specific morbidity 

for almost all diagnoses. There was a reduction in risk for 

liver morbidity for patients exposed to antibiotics (n= 27 756) 

between 0.55 (0.44-0.70) and 0.77 (0.73-0.80). Conclusion:


Several frequently used drug classes in patients with chronic 

liver disease are associated with reduced all-cause mortality, 

and liver-specific morbidity and mortality. Statin, ACEi and 

ARB use was associated with lower risk of total mortality, liver-specific 

mortality and liver-specific morbidity. ASA use was 

associated with reduced liver-specific mortality, and NSAID, 

SSRI and antibiotics exposure was associated with a reduction 

in risk for liver morbidity. 

Disclosures: 

Anders Ekbom - Advisory Committees or Review Panels: Centocor, Schering-Plough, 

Centocor, Schering-Plough, Centocor, Schering-Plough, Centocor, 

Schering-Plough; Speaking and Teaching: Astra Zeneca, Astra Zeneca, Astra 

Zeneca, Astra Zeneca 

The following authors have nothing to disclose: Knut Stokkeland, Christine 

Takami Lageborn, Jonas Höijer, Matteo Bottai, Per Stål, Karin Söderberg Löfdal 

that becomes significantly more accurate than its 2 composite 

tests, and solves discrepancies between these tests. The 

combined test that associates elastography and a blood test, 

used with its detailed fibrosis classification, should become the 

reference. 

Disclosures: 





Isabelle Fouchard-Hubert - Speaking and Teaching: JANSSEN, BMS, GILEAD, 

ABBVIE 


The following authors have nothing to disclose: Jerome Boursier, Oberti Frederic 

117 

Evaluation of EASL recommendation for the non-invasive 

diagnosis of liver fibrosis in chronic hepatitis C 

Jerome Boursier 1,3 , Victor de Ledinghen 2 , Vincent Leroy 4 , Oberti 

Frederic 1,3 , Isabelle Fouchard-Hubert 1,3 , Paul Cales 1,3 ; 1 Hepato-gastroenterology 

Department, University Hospital, Angers, 

France; 2 Hepatology Department, University Hospital, Bordeaux, 

France; 3 HIFIH Laboratory EA3859, University, Angers, France; 

4 Hepato-Gastroenterology Department, University Hospital, Grenoble, 

France 

Introduction. The EASL has proposed the first recommendations 

about non-invasive testing for the evaluation of liver disease 

severity. For chronic hepatitis C (CHC), it is proposed to associate 

transient elastography (TE) with a blood test (BT), and to 

accept the diagnosis if both are in agreement. We aimed to 

validate this rule and to improve it by using a combined fibrosis 

test that simultaneously associates both BT and TE (BET). Methods. 

679 CHC patients with liver biopsy, TE, and a patented BT 

were included. The primary outcome was severe fibrosis (Metavir 

F≥3, prevalence: 31.9%). Diagnostic cut-off for this target 

was evaluated according to the maximum Youden index or the 

threshold between the 2 classes F2±1 and F3±1 of a detailed 

fibrosis classification including 6 fibrosis classes. Results. 

AUROC was: BT: 0.829, TE: 0.846, BET: 0.882 (p


Hans Huber - Consulting: MiNA Therapeutics; Stock Shareholder: Merck & Co 

Robert Habib - Board Membership: MiNA Therapeutics Ltd; Management Position: 

MiNA Therapeutics Ltd; Stock Shareholder: MiNA Therapeutics Ltd 

Pål Sætrom - Patent Held/Filed: MiNA Therapeutics, Ltd; Stock Shareholder: 

MiNA Therapeutics, Ltd 

Nagy Habib - Board Membership: EMcision Limited, Omnicyte Limited, Apterna 

Limited, MiNA Therapeutics Ltd; Patent Held/Filed: EMcision Limited, Omnicyte 

Limited; Stock Shareholder: EMcision Limited, Omnicyte Limited, Apterna Limited, 

MiNA Therapeutics Ltd 

The following authors have nothing to disclose: Kai-Wen Huang, Anjaneyulu 

Muragundla, Aravindakshan Jayaprakash, Prashant Vadnal, John Rossi 

119 

Fibrosis is not just fibrosis - Extracellular matrix turnover 

is markedly different in two types of viral hepatitis, suggesting 

different fibrotic representations 

Mette J. Nielsen 1,2 , Morten A. Karsdal 1 , Konstantin Kazankov 3 , 

Aleksander Krag 2 , Diana J. Leeming 1 , Jacob George 4 , Henning 

Gronbaek 3 , Detlef Schuppan 5,6 ; 1 Nordic Bioscience A/S, Herlev, 

Denmark; 2 Department of Gastroenterology and Hepatology, 

Odense University Hospital, University of Southern Denmark, 

Odense, Denmark; 3 Department of Medicine V, Aarhus University 

Hospital, Aarhus, Denmark; 4 Storr Liver Unit, Westmead Millennium 

Institute, Westmead Hospital and University of Sydney, 

Sydney, NSW, Australia; 5 Institute of Translational Immunology, 

University Medical Center, Mainz, Germany; 6 Division of Gastroenterology, 

Beth Israel Deaconess Medical Center, Harvard Medical 

School, Boston, MA 

Background and aim: Fibrosis is the result of a dysregulated 

tissue remodelling leading to excessive and abnormal accumulation 

of extracellular matrix (ECM). The developmental pattern 

of fibrosis depends on the underlying aetiology causing the 

fibrosis such as; portal-portal, portal-central, or central-central 

septa, albeit the molecular composition and turnover remain 

to be investigated. In this study we investigated the level of 

ECM remodelling in two seemingly similar types of viral hepatitis, 

namely hepatitis B and C. Methods: In a cross-sectional 

study design, specific protein fragments of matrix metalloprotease 

degraded type I, III, IV and VI collagen (C1M, C3M, 

C4M, C6M) and type III and IV collagen formation (Pro-C3 

and P4NP7S) were assessed in plasma from 403 chronic 

hepatitis C patients and 197 chronic hepatitis B patients by 

specific ELISAs. The grade of inflammation and fibrosis was 

scored according to the Metavir Activity and Fibrosis Scoring 

systems based on liver biopsy. Results: Plasma levels of 

P4NP7S, C3M, C4M, and C6M were significantly elevated 

in HBV compared to HCV (p


121 

Objective Determination of Hepatitis B Phenotype using 

Biochemical and Serological Markers: Immune Tolerant, 

Immune Active, Inactive Carrier and Indeterminant Status 

Adrian M. Di Bisceglie 1 , Manuel Lombardero 2 , Jeffrey Teckman 1 , 

Lewis R. Roberts 3 , Harry L. Janssen 4 , Steven H. Belle 2 , Jay H. Hoofnagle 

5 ; 1 Saint Louis University, St. Louis, MO; 2 University of Pittsburgh, 

Pittsburgh, PA; 3 Mayo Clinic, Rochester, MN; 4 University of 

Toronto, Toronto, ON, Canada; 5 NIH, Bethesda, MD 

Background: HBV infection is frequent world-wide and may 

result in progressive liver disease or HCC. Effective therapies 

are available but treatment is usually reserved for patients (pts) 

with selected biochemical and serological profiles (phenotypes). 

Currently, criteria for defining HBV phenotype are not 

standardized and are based upon expert opinion rather than 

medical evidence. Aim: To determine the distribution of phenotypes 

in a large cohort of North American pts with chronic 

HBV infection using standardized definitions and calculation. 

Methods: Epidemiologic, demographic, biochemical and virologic 

features of pts enrolled into the HBRN Cohort Study at 19 

US and 1 Canadian site were analyzed, assigning pts into 1 

of 4 phenotypes: immune tolerant (IT), chronic hepatitis B with 

(CHB e+) or without HBeAg (CHB e-) or inactive carrier (IC) 

based upon baseline HBV DNA and ALTs. Cut-off HBV DNA 

values used to define phenotypes were 10 4 IU/ml for e-negative 

and 10 5 for e-positive pts. ALT cut-offs were analyzed using 

either fixed values (30 U/L for men, 20 U/L for women) or 

using each local lab ULN. Pts not fitting into a phenotype were 

designated “Indeterminant”. Independently, local investigators 

assigned a phenotype at baseline based on available laboratory 

values and clinical impression. Results: 1398 adults (51% 

male, 71% Asian) had data needed to determine phenotype. 

The table shows the distribution of phenotypes calculated by 

two methods of assessing ALT and physician assessment. Only 

moderate agreement was found between clinician-determined 

and calculated phenotype using fixed ALTs (Kappa 0.39, 95% 

CI 0.36-0.42). Of note, only 13% of pts were designated Indeterminant 

by clinicians compared to 20-40% by computer calculation. 

Using the Fixed ALT groups for comparison, IT was 

most distinct, being generally younger, more frequently Asian 

and female. The most frequent clinical phenotype identified 

was Indeterminant, the majority of whom (83%) had elevated 

ALT values despite low levels of HBV DNA, not apparently 

associated with higher BMI, alcohol consumption or HBV genotype. 

Clinician estimates often differed from the calculated 

phenotype. Conclusions: The clinical significance of HBV phenotypes 

using these standardized definitions requires further 

study based on long term follow up of these patient cohorts but 

there is clearly a need to re-examine categorization of pts with 

chronic HBV infection to more accurately predict their outcomes 

and need for therapy. 

Disclosures: 

Adrian M. Di Bisceglie - Advisory Committees or Review Panels: Gilead, AbbVie, 

Novartis, Bayer, BTG; Grant/Research Support: Gilead, AbbVie 

Jeffrey Teckman - Consulting: Dicerna, Isis Pharmaceuticals, Vertex, Proteostasis, 

Genkyotex, The Alpha-1 Project, Retrophin, RxCelerate, Velgene; Grant/ 

Research Support: Alnylam, Arrowhead, Alpha-1 Foundation, Gilead 

Lewis R. Roberts - Grant/Research Support: Bristol Myers Squibb, ARIAD Pharmaceuticals, 

BTG, Wako Diagnostics, Inova Diagnostics, Gilead Sciences, Five 

Prime Therapeutics 

Harry L. Janssen - Consulting: AbbVie, Bristol Myers Squibb, GSK, Gilead Sciences, 

Innogenetics, Merck, Medtronic, Novartis, Roche, Janssen, Medimmune, 

ISIS Pharmaceuticals, Tekmira; Grant/Research Support: AbbVie, Bristol Myers 

Squibb, Gilead Sciences, Innogenetics, Merck, Medtronic, Novartis, Roche, 

Janssen, Medimmune 

The following authors have nothing to disclose: Manuel Lombardero, Steven H. 

Belle, Jay H. Hoofnagle 

122 

Length of antiviral therapy and cirrhosis are key factors 

in the development of hepatocellular carcinoma among 

U.S. veterans with chronic hepatitis B 

Gina Choi, Marina Serper, David E. Kaplan, Kimberly A. Forde; 

Gastroenterology, University of Pennsylvania, Philadelphia, PA 

Background: The risk of hepatocellular carcinoma (HCC) and 

the effect of anti-viral therapy in U.S. populations with chronic 

hepatitis B (HBV) is poorly defined. Aim: To examine the incidence 

and impact of anti-viral therapy on the development of 

HCC among a national cohort of veterans with chronic HBV. 

Methods: A retrospective cohort study using the VA Corporate 

Data Warehouse was performed between 1999-2013. The 

primary exposure variable was antiviral therapy, defined as 

≥6 months of therapy with oral nucleosides. Additional covariates 

such as demographics and clinical variables such as cirrhosis, 

alcohol abuse, dyslipidemia, hepatitis C (HCV), and 

HIV were obtained. Multivariable Cox proportional hazard 

models were used to evaluate associations between exposures 

and the primary outcome of HCC. Results: A total of 26,704 

veterans had a HBsAg+ result; N=9001 were confirmed to 

have chronic HBV. The median follow-up time was 7.6 years 

(IQR 4.2–10.9). Patients were predominantly male (96%), 

white (45%) and middle aged (M=51, SD 11). A total of 20% 

had HCV, 7% had HIV, and 23% had alcohol abuse. The 

prevalence of dyslipidemia was 52%; 13% had cirrhosis. The 

overall incidence of HCC was 5.6 per 1000 person-years; 

26 per 1000 person-years with cirrhosis and 1.9 per 1000 

person-years without cirrhosis. The median exposure time of 

antiviral therapy was 3.6 years (IQR 1.4-7.1). A total of 38% 

(N=3333) received antiviral therapy for ≥6 months; 23% 

received ≥3 years and 16% received therapy for ≥5 years. 

In multivariable models adjusted for age, race, cirrhosis, alcohol 

abuse, and dyslipidemia, antiviral therapy ≥5 years was 

associated with decreased incidence of HCC (HR 0.77, 95% 

CI 0.60-0.98, p=0.03), whereas ≥4 years of antiviral therapy 

was not protective (HR 1.04, CI 1.03-1.05). Antiviral therapy 

for ≥5years was associated with a decreased incidence of 

HCC among cirrhotics (HR 0.56, CI 0.41-0.78, p


123 

Serum Alanine Aminotransferase (ALT) and Hepatitis 

B Virus (HBV) DNA Flares In Pregnant and Postpartum 

Women With Chronic Hepatitis B (CHB) 

Christine Y. Chang 1 , Natali Aziz 2 , Huy N. Trinh 3 , Mindie H. 

Nguyen 1 ; 1 Division of Gastroenterology and Hepatology, Stanford 

University Medical Center, Palo Alto, CA; 2 Obsterics and Gynecology, 

Stanford University School of Medicine, Stanford, CA; 3 San 

Jose Gastroenterology, San Jose, CA 

Background: During pregnancy, alterations in the immune system 

allow the mother to tolerate the semiallogenic fetus, but 

their effects on CHB activity are poorly understood. Methods: 

To examine flares in HBV DNA (≥1 log increase from baseline) 

and/or ALT (≥2xULN or baseline) during pregnancy and 6 

months postpartum, we performed a retrospective study of 88 

pregnancies in 74 CHB expectant mothers who were not on 

anti-HBV therapy for ≥1 year prior to pregnancy at 2 U.S. 

centers. Results: Most were Asian (93%), with a mean age 

of 32±4, median gravida of 2 (IQR: 1-3), and median parity 

of 0 (IQR: 0-1). Mean log HBV DNA was 2.0±0.8 IU/mL at 

baseline in low HBV DNA mothers (≤2000 IU/mL, n=33), and 

6.1±2.0 in high HBV DNA mothers (>2000 IU/mL, n=55). At 

baseline, low HBV DNA mothers had lower ALT (≤x2ULN: 82% 

vs. 47%, p=0.001) and were more likely to be HBeAg- (90% 

vs. 44%, p


Disclosures: 

The following authors have nothing to disclose: Prabhu P. Gounder, Lisa Bulkow, 

Mary Snowball, Susan Negus, Philip R. Spradling, Brian J. McMahon 

126 

Relationship between hepatocellular carcinoma development 

and serum viral markers in chronic hepatitis B 

patients who achieved undetectable serum HBV DNA 

while on long-term nucleoside analogue therapy 

Ka-Shing Cheung 1 , Wai-Kay Seto 1 , Danny Wong 2 , Ching-Lung 

Lai 1 , Man-Fung Yuen 1 ; 1 Medicine, Queen Mary Hospital, The University 

of Hong Kong, Hong Kong, Hong Kong; 2 Queen Mary 

Hospital, Hong Kong, Hong Kong 

Background: Hepatitis B surface antigen (HBsAg) and hepatitis 

B core-related antigen (HBcrAg) are risk factors for the 

development of hepatocellular carcinoma (HCC). Linearized 

HBsAg (HQ-HBsAg) is a more sensitive assay to measure 

HBsAg level. However, their roles in predicting HCC development 

are unknown when there is profound viral suppression 

by nucleos(t)ide analogues (NA). Methods: Seventy-six chronic 

hepatitis B (CHB) patients who developed HCC in 2007-2014 

despite undetectable serum HBV DNA level after at least oneyear 

NA therapy were recruited. 152 CHB patients without 

HCC were recruited as controls. They were matched by age, 

gender, HBeAg status, cirrhosis status, undetectable serum HBV 

DNA and duration of NA therapy with the 76 HCC patients in 

a 2:1 ratio. Serum HBsAg, HQ-HBsAg and HBcrAg levels were 

analysed and compared between the two groups. Results: A 

statistically significant difference in the HBcrAg level was noted 

between the HCC group and non-HCC groups (10.2 and 1.7 

kU/mL respectively, p=0.005), but was not observed in HBsAg 

or HQ-HBsAg levels. The area under receiver operating characteristic 

curve (AUROC) was 0.61 (95% CI: 0.54-0.69) with a 

negative predictive value (NPV) of 77.0% when a cutoff value 

of HBcrAg level ≥7.8 kU/mL was used. The odds ratio of HCC 

development was 3.27 (95% CI: 1.84-5.80). In the subgroup 

analysis for non-cirrhotic patients, a statistically significant 

difference in the HBcrAg level was noted between the HCC 

group and non-HCC groups (10.2 and 1.0 kU/mL respectively, 

p=0.001). The AUROC was 0.70 (95% CI: 0.58-0.81) with a 

NPV of 80.6% when a cutoff value of HBcrAg level ≥7.9 kU/ 

mL was used. The odds ratio of HCC development was 5.95 

(95% CI: 2.35-15.07). Conclusion: HBcrAg (but not HBsAg or 

HQ-HBsAg) can predict HCC risk in CHB patients who achieve 

undetectable serum HBV DNA while receiving NA therapy. 

Future prospective studies are warranted to further define the 

role of HBcrAg level in this group of patients. 

Disclosures: 

Wai-Kay Seto - Advisory Committees or Review Panels: Gilead Science; Speaking 

and Teaching: Bristol-Myers Squibb 

Ching-Lung Lai - Advisory Committees or Review Panels: Bristol-Myers Squibb, 

Gilead Sciences Inc; Consulting: Bristol-Myers Squibb, Gilead Sciences, Inc; 

Speaking and Teaching: Bristol-Myers Squibb, Gilead Sciences, Inc 

Man-Fung Yuen - Advisory Committees or Review Panels: GlaxoSmithKline, 

Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, 

GlaxoSmithKline, Bristol-Myers Squibb, Pfizer, GlaxoSmithKline, Bristol-Myers 

Squibb, Pfizer; Grant/Research Support: Roche, Bristol-Myers Squibb, 

GlaxoSmithKline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmith- 

Kline, Gilead Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead 

Science, Roche, Bristol-Myers Squibb, GlaxoSmithKline, Gilead Science 

The following authors have nothing to disclose: Ka-Shing Cheung, Danny Wong 

127 

A New Clinically-Based Staging System for Distal Cholangiocarcinoma 

Maria E. Lozada 1 , Jonggi Choi 1 , Roongruedee Chaiteerakij 1,2 , 

Ju Dong Yang 1 , Nasra H. Giama 1 , Steven Alberts 3 , Gregory J. 

Gores 1 , Lewis R. Roberts 1 ; 1 Division of Gastroenterology and 

Hepatology, Mayo Clinic College of Medicine, and Mayo Clinic 

Cancer Center, Rochester, MN; 2 Department of Medicine, Faculty 

of Medicine, Chulalongkorn University and King Chulalongkorn 

Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand; 

3 Division of Medical Oncology, Mayo Clinic College of Medicine, 

and Mayo Clinic Cancer Center, Rochester, MN 

Background: Clinical predictors of survival for each subtype of 

cholangiocarcinoma, intrahepatic, perihilar and distal (dCCA) 

are limited. Currently available staging systems can be used to 

predict prognosis for resectable dCCA but are not relevant to 

patients with unresectable disease. Based on these limitations, 

we aimed to construct a clinical staging system that stratifies 

dCCA patients. Methods: We identified 170 treatment-naïve 

dCCA patients seen at Mayo Clinic, Rochester from January 

1, 2000 through May 31, 2014. Data were retrospectively 

abstracted from the electronic medical record. Overall median 

survival (MS), the primary endpoint, was estimated using the 

Kaplan-Meier method and compared using the log-rank test. 

Cox-proportional hazards analysis was used to identify predictors 

of survival. Performance of the staging system was assessed 

using concordance index. Results: Of the 170 dCCA patients, 

93 (58%) were male, the mean age was 67 years, and 19 

(12%) had primary sclerosing cholangitis. The MS of the entire 

cohort was 17.0 months (95% confidence interval [CI]: 14.0- 

20.5). Baseline ECOG performance status, post-stenting CA 

19-9 and disease extent were significantly associated with survival 

and further incorporated into a 4-tier staging system. A 

total of 143 patients were classified into the new staging system. 

Overall MS for patients in Stage I, Stage II, Stage III and 

Stage IV were 46.8, 14.3, 8.1, and 3.4 months, with hazard 

ratios (95% CI) of 1.0 (ref.), 1.9 (1.0-3.4), 4.1 (2.3-7.0), and 

9.7 (5.1-18.5), respectively. Concordance index for the new 

staging system was 0.73. Conclusion: We have developed a 

new staging system based on non-operative clinical variables 

that classifies dCCA patients into 4 distinct stages with homogeneous 

survival. Given the small cohort, validation to confirm 

these findings is crucial. However, this staging system could 

potentially be a key prognostic tool for better stratification of 

patients enrolled in clinical trials of novel therapies for dCCA. 

Disclosures: 

Gregory J. Gores - Advisory Committees or Review Panels: Conatus 



Prime Therapeutics


The following authors have nothing to disclose: Maria E. Lozada, Jonggi Choi, 

Roongruedee Chaiteerakij, Ju Dong Yang, Nasra H. Giama, Steven Alberts 

128 

Trends in the burden of cirrhosis and hepatocellular carcinoma 

by underlying liver disease in US Veterans from 

2001-2013 

Lauren A. Beste 2 , Steven Leipertz 2 , Pamela Green 2 , Jason A. Dominitz 

2,1 , David Ross 3 , George N. Ioannou 2,1 ; 1 University of Washington, 

Seattle, WA; 2 Veterans Affairs Puget Sound Healthcare 

System, Seattle, WA; 3 Office of Public Health, Department of Veterans 

Affairs, Washington, D.C., DC 

Background & Aims Cirrhosis and hepatocellular carcinoma 

(HCC) are predicted to increase in the U.S. but the accuracy 

of prior forecasts and the contributions from various etiologies 

remain unclear. We aimed to determine the burden of 

cirrhosis and HCC according to underlying etiology from 

2001-2013. Methods We developed a national retrospective 

cohort of Veterans Affairs (VA) patients diagnosed with cirrhosis 

(n=129,998) or HCC (n=21,326) from 2001-2013. 

We used laboratory results, ICD-9 codes, and biometrics to 

identify underlying etiologies. Results In 2013, VA provided 

care to 5,720,614 individuals, of whom 60,553 (1.06%) had 

cirrhosis and 7,670 (0.13%) had HCC. Hepatitis C virus (HCV) 

was present in an increasing proportion of cirrhosis and HCC 

between 2001-2013, reaching 48% of cirrhosis cases and 

deaths and 67% of HCC cases and deaths by 2013. Cirrhosis 

prevalence nearly doubled from 2001-2013 (664 to 1058 

per 100,000 enrollees), driven by HCV and nonalcoholic fatty 

liver disease (NAFLD). Cirrhosis incidence ranged from 159 

to 193 per 100,000 patient-years. Deaths in patients with 

cirrhosis increased from 83 to 126 per 100,000 patient-years, 

largely driven by HCV. HCC incidence increased 2.5-fold from 

17 to 45 per 100,000 patient-years. HCC mortality tripled 

from 13 to 37 per 100,000 patient-years, driven overwhelmingly 

by HCV with much smaller contributions from NAFLD and 

alcoholic liver disease. Conclusions Cirrhosis prevalence and 

mortality and HCC incidence and mortality increased from 

2001-2013 driven by HCV with a smaller contribution from 

NAFLD. If current trends continue, cirrhosis prevalence will 

peak in 2021. Healthcare systems will need to accommodate 

rising numbers of patients with cirrhosis and HCC. 

INCIDENCE OF HCC IN THE VETERANS AFFAIRS HEALTHCARE 

SYSTEM 2002-2012, BY UNDERLYING LIVER DISEASE 

Disclosures: 

Jason A. Dominitz - Employment: Department of Veterans Affairs; Grant/Research 

Support: Gilead Pharmaceuticals 

The following authors have nothing to disclose: Lauren A. Beste, Steven Leipertz, 

Pamela Green, David Ross, George N. Ioannou 

129 

Multi-platform analysis of Telomerase Reverse Transcriptase 

(TERT) gene alterations in Hepatocellular Carcinoma 

(HCC) 

Renumathy Dhanasekaran 1 , Kyle Covington 2 , Jennifer Watt 2 , 

Andrew D. Cherniack 3 , Daniel R. O’Brien 4 , Ju-Seog Lee 5 , Chad 

Creighton 2 , Donna M. Munzy 2 , Lewis R. Roberts 1 , David A. 

Wheeler 2 ; 1 Gastroenterology and Hepatology, Mayo Clinic, Rochester, 

MN; 2 Baylor College of Medicine, Houston, TX; 3 Broad 

Institute of MIT and Harvard, Cambridge, MA; 4 Mayo Clinic, Rochester, 

MN; 5 The University of Texas MD Anderson Cancer Center, 

Houston, TX 

Background: Telomerases are enzymatic protein complexes 

which maintain telomere length and play an important role in 

cancer cell immortalization. Mechanisms of TERT gene activation 

and clinical associations of TERT gene alterations in 

HCC are not completely understood. Methods: We used data 

from TCGA analysis of HCC primary tumor from 192 patients. 

Germline DNA from blood or benign surrounding liver was 

used as reference. Mutational, copy number, mRNAseq and 

viral genomic integration analyses were performed. Results: 

The TERT gene was found to be significantly overexpressed 

in HCC tumor tissue when compared to benign surrounding 

liver (pA (C228T) and 1,295,250 G>A (C250T)) were found. 

The majority of tumors had the C288T mutation (92.1%) and 

six (7.9%) tumors had the C250T mutation. Patients with TERT 

promoter mutation were older (p=0.03) and predominantly 

male (p=0.02). Patients with TERT promoter mutation were 

more likely to have hepatitis C than no mutation (33% vs 10%; 

p=0.02). In contrast, patients with TERT promoter mutation 

were less likely to have hepatitis B than no mutation (14% vs. 

27%; p=0.05). Also, patients with TERT promoter mutation 

had worse survival than patients without the mutation (3 year 

survival 59% vs. 39%; p=0.02). Further, we report here the first 

case of a germline mutation in the TERT promoter in HCC in a 

30 year old male who did not have any risk factors for HCC. 

Copy number analyses showed TERT gene amplification in 

7% of tumor samples; TERT was the fourth most common gene 

found to be amplified in HCC. There was significant correlation 

between TERT gene amplification and TERT mRNA overexpression. 

In the subset of patients with HBV infection, the integration 

of HBV viral genome into TERT promoter region was identified 

in 13.6% (6/44) of patients and this was the most common site 

of recurrent HBV integrations. Conclusion: This is the first comprehensive 

multi-platform analysis of the various mechanisms of 

TERT gene alterations in HCC and their association with patient 

outcomes. TERT promoter mutations were found to be the most 

common somatic mutation in HCC and were associated with 

poor survival. We identify TERT promoter somatic mutations, 

TERT gene amplification and HBV integrations into the TERT 

promoter region as various mechanisms of TERT gene alterations. 

Also, we report here the first case of germline mutation 

of the TERT promoter gene in HCC, possibly resulting in an 

inherited predisposition to HCC.


Disclosures: 

Andrew D. Cherniack - Grant/Research Support: Bayer AG 




The following authors have nothing to disclose: Renumathy Dhanasekaran, Kyle 

Covington, Jennifer Watt, Daniel R. O’Brien, Ju-Seog Lee, Chad Creighton, 

Donna M. Munzy, David A. Wheeler 

130 

Targeting the TGF-β Pathway Increases Patient Survival 

for Hepatocellular Cancer 

Jian Chen 1 , Jiun-Sheng Chen 1 , Wilma Jogunoori 1 , Young Jin Gi 1 , 

Yun Seong Jeong 1 , Xiaoping Su 1 , Asif Rashid 1 , Bibhuti Mishra 2 , 

Jon White 2 , Milind Javle 1 , Marta L. Davila 1 , John R. Stroehlein 1 , 

Xuemei Wang 1 , Jeffrey S. Morris 1 , Rehan Akbani 1 , Lopa Mishra 1 ; 

1 The University of Texas MD Anderson Cancer Center, Houston, 

TX; 2 Institute of Clinical Research, Veterans Affairs Medical Center, 

Washington DC, DC 

Liver cancers that include hepatocellular cancer (HCC) and 

intrahepatic cholangiocarcinoma are among the most challenging 

and lethal malignancies with a degree of complexity seldom 

seen in other cancers. Patients with HCC have a poor survival 

of 9-11 months. However, key driver pathways and identifying 

specific populations responsive to targeted therapeutics have 

remained elusive. Recent clinical studies show that targeting 

TGF-β in patients with high TGF-β level improves survival up 

to 21 months. Therefore, we hypothesized that genomic, transcriptomic 

profiles coupled with mouse mutants and functional 

analyses may identify specific patients with HCC with a high 

response rate to targeting TGF-β and present an attractive new 

treatment strategy. Methods and Results: (1) We analyzed the 

transcriptome of 488 hepatocellular cancers and screened for 

mutations in the TGF-β pathway in 202 HCCs from The Cancer 

Genome Atlas (TCGA). We found aberrant TGF-β superfamily 

in 72% of HCC with mutations in 38%, the greatest number 

being for the Smad adaptor β2SP (6%). (2) Increased levels of 

TGF-β genes were designated as an “activated” signature that 

is associated with hepatic fibrosis. Decreased levels of TGF-β 

genes were defined as an “inactivated” signature, which was 

associated with the loss of TGF-β tumor suppressor function. 

HCCs characterized by the “inactivated” TGF-β signature were 

associated with a significantly poorer survival particularly in 

early stage HCCs, compared to HCCs with the “activated” 

TGF-β signature (p=0.0027). (3) The TGF-β pathway is statistically 

significantly associated with DNA repair genes. (4) 

We performed (80x) whole-genome sequence analysis of eight 

additional HCCs to define the role of TGF-β in their development 

and characterize a potential novel “driver mutations” in 

HCV- and alcohol-associated HCC. (5) Further functional analyses 

were performed in mouse mutants in the TGF-β pathway 

that spontaneously develop HCC, patient derived xenografts, 

and human cell lines. Increased levels of TGF-β associated-E3 

ligases, including KEAP1, and PJA1 were found in 20% of 

HCCs. Targeting these E3 ligases revealed a strong response 

using patient derived xenografts. Conclusions: The molecular 

signatures of the TGF-β pathway we characterize here appear 

to have prognostic significance. The additional association 

with the DNA repair pathway supports new approaches to 

developing biomarkers. TGF-β and TGF-β-associated-E3 ligases 

are activated in distinct groups of liver cancer patients. It may 

be possible to identify subsets of patients, defined by the activation 

or inactivation of TGF-β signaling, who may respond more 

effectively to specific treatments. 

Disclosures: 

The following authors have nothing to disclose: Jian Chen, Jiun-Sheng Chen, 

Wilma Jogunoori, Young Jin Gi, Yun Seong Jeong, Xiaoping Su, Asif Rashid, 

Bibhuti Mishra, Jon White, Milind Javle, Marta L. Davila, John R. Stroehlein, 

Xuemei Wang, Jeffrey S. Morris, Rehan Akbani, Lopa Mishra 

131 

A Study for the Risk Factors of Hepatocellular Carcinoma 

in Cirrhotic Patients with Chronic Hepatitis B 

Yuanqing Zhang 1 , Lijun Peng 1,2 , Yirong Cao 1 , Zhiping Zeng 1 , 

Yujing Wu 1 , Hong Shi 1 , Shiyao Chen 1 , Jiyao Wang 1 , Scott L. 

Friedman 1,3 , John J. Sninsky 1,4 , Jinsheng Guo 1 ; 1 Division of Digestive 

Diseases, Zhong Shan Hospital, Fu Dan University, Shanghai, 

China; 2 Department of Gastroenterology, Linyi People’s Hospital, 

Linyi, China; 3 Division of Liver Diseases,, Mount Sinai Hospital, 

Icahn School of Medicine at Mount Sinai, New York, NY; 4 Celera/Quest 

Diagnostics, Alameda, CA 

Background and Aims: Chronic hepatitis B virus infection and 

cirrhosis are important risk factors for hepatocellular carcinoma 

(HCC), however the HCC risk can vary widely among individuals. 

The aim of this study was to identify the clinical factors 

and host genetic single nucleotide polymorphisms (SNPs) that 

are associated with HCC risk in cirrhotic patients with CHB. 

Methods: Data from 949 Chinese Han patients with chronic 

HBV infection were analyzed by single and multivariate logistic 

regression analysis in cirrhotic patients without (n = 281) and 

with HCC (n = 434) to identify the clinical factors associated 

with HCC risk. In a parallel study, DNA was extracted from 

879 chronic HBV patients including non-cirrhotic HBV carriers 

and CHB patients (n = 234), cirrhotic patients without (n = 

257) and with HCC (n = 388) for genotyping of ten candidate 

SNPs using polymerase chain reaction-ligase detection 

reaction method. The correlations between the candidate SNPs 

and HCC risk were analyzed by chi-square test followed by 

logistic regression analysis. Results: 1. Ineffective antiviral treatment 

(OR=5.2), fatty liver (OR=3.4), family history of HCC 

(OR=2.3), drinking history (OR=2.2), and age≥50 (OR=1.8) 

were independent risk factors for HCC in cirrhotic patients with 

CHB. Longer HBV infection increased the HCC risk whereas 

sustained virologic suppression significantly lowered HCC 

risk compared to those without enough response or untreated 

patients, especially in patients with decompensated cirrhosis. 

Even after achieving viral suppression, 36.5% (62/170) of cirrhotic 

patients with CHB still developed HCC. Fatty liver, family 

history of HCC and HBV infection were significantly associated 

with the HCC development in these patients. 2. After adjusted 

for influencing factors, TLR4 rs11536889 was found to be a 

protective factor (OR for CC vs. GG+GC or GG =0.4 and 

0.6, respectively) whereas SPP1 rs2853744 (OR for TT vs. GG 

and GT+TT vs. GG =1.9 and 3.1, respectively) and AP3S2 

rs2290351 (OR for GA+AA or GA vs. GG =2.5 and 2.9, 

respectively) were risk factors of HCC in cirrhotic patients with 

CHB. In cirrhotic patients with CHB and drinking history, MLEC 

rs7976497 (OR for TT vs. CC=2.8, CT+TT vs. CC=3.6) and 

SOCS3 rs4969168 (OR for GG vs. AA=2.4) were found to be 

risk factors of HCC. Conclusion: Ineffective antiviral treatment, 

fatty liver, family history of HCC, drinking history and age≥50 

are risk factors for HCC. Sustained suppression of HBV does 

not remove the risk of HCC. Specific host genetic factors may 

impact on HCC development in cirrhotic patients with CHB 

including a protective SNP in TLR4, and risk SNPs in SPP1, 

AP3S2, MLEC, and SOCS3.


Disclosures: 











John J. Sninsky - Consulting: Roche Diagnostics; Employment: CareDx 

The following authors have nothing to disclose: Yuanqing Zhang, Lijun Peng, 

Yirong Cao, Zhiping Zeng, Yujing Wu, Hong Shi, Shiyao Chen, Jiyao Wang, 

Jinsheng Guo 

132 

Diabetes, HBV infection and smoking are independent 

risk factors for developing hepatocellular carcinoma 

on non-fibrotic liver in the NoFLIC French multicenter 

case-control study. 

Jean-Frédéric Blanc 1 , Adelaide Doussau 2 , Marie-Quitterie Picat 2 , 

Aline Maillard 2 , Caroline Bouyssou 1 , Charlotte E. Costentin 3 , Olivier 

Rosmorduc 4 , Isabelle Rosa 5 , Luc Letenneur 2 , Karen Leffondre 6 , 

Jessica Zucman-Rossi 6 , Marianne Ziol 7 ; 1 Hepatology and Digestive 

Oncology Unit, CHU bordeaux, Bordeaux, France; 2 ISPED, 

Bordeaux, France; 3 Hepatology, Hopital Henri Mondor, Creteil, 

France; 4 Heptology, Hopital saint-Antoine, Paris, France; 5 Hepatology, 

CHI Créteil, Créteil, France; 6 INSERM U1162, Paris, France; 

7 Pathologie, Hopital Jean Verdier, Bondy, France 

risk factors for developing nfHCC. Conclusion NoFLIC is the 

first case-control study dedicated to the study of factors associated 

with the occurrence of HCC in non-fibrotic liver (F0/F1). 

The first analysis shows a significant association between metabolic 

syndrome, diabetes, HBV infection and smoking with the 

occurrence of HCC in non-fibrotic liver suggesting an important 

role of these factors in liver carcinogenesis. 

Disclosures: 

Jean-Frédéric Blanc - Advisory Committees or Review Panels: Lilly / Imclone, 

Merck Serono, Sanofi; Speaking and Teaching: Bayer, BMS, Roche, Abbvie 

Olivier Rosmorduc - Advisory Committees or Review Panels: Syrtex, IPSEN; 

Speaking and Teaching: Bayer 

Jessica Zucman-Rossi - Advisory Committees or Review Panels: Astellas, Celgene; 

Consulting: Pfizer; Grant/Research Support: Integragen; Speaking and Teaching: 

Bayer, Lilly 

Marianne Ziol - Grant/Research Support: Echosens 

The following authors have nothing to disclose: Adelaide Doussau, Marie-Quitterie 

Picat, Aline Maillard, Caroline Bouyssou, Charlotte E. Costentin, Isabelle 

Rosa, Luc Letenneur, Karen Leffondre 

Introduction The occurrence of HCC in non-fibrotic liver (nfHCC) 

is a rare event (5% of HCC patients) and factors contributing to 

their onset remain poorly understood. The aim of the case-control 

exploratory study NoFLIC was to identify factors involved 

in hepatocarcinogenesis in non-fibrotic liver. Patients and 

methods NoFLIC is a French multicenter case-control study that 

included consecutive patients who developed HCC in a non-fibrotic 

liver, F0 or F1, proven by histological analysis. Controls 

were matched for age and gender, they were recruited in 

people with gastro-intestinal screening and normal endoscopy, 

or in rheumatology departments. Clinical data were collected 

including the parameters of the metabolic syndrome defined 

according to the NCEP-ATP3 criteria, viral serology, quantification 

of alcohol and tobacco use. Data were taken from the 

patient’s medical record, the results of blood test performed for 

this study and a specific epidemiologic survey. Logistic regressions 

adjusted for age, sex and geographical origin, were performed 

to determine the association of each of the risk factors 

with the status of the patient (case / control). A multivariate 

analysis was performed to study the effect of significant risk 

factors. Results A total of 109 nfHCC (cases) and 163 controls 

were included in 12 hospitals between 2010 and 2013. 

Mean age of the nfHCC patients was 64.4 years, with 68.8% 

male. Active HBV infection was detected in 6 cases (OR 11.4, 

95% 1.18-110.84, p=0.035). History of alcohol abuse was 

more frequent in nfHCC (OR 2.13, 95% 1.09-4.16 p=0.026). 

A metabolic syndrome was identified in 28% of the patients 

developing nfHCC (OR 3.3; 95% CI 1.62-6.82 p=0.001). 

Among the components of metabolic syndrome, treated diabetes 

or fasting glucose ≥ 6.1mmol/l was significantly associated 

with nfHCC occurrence (OR 3.08; 95% CI 1.67-5.69 

p=0.0002), while there was no significant association with 

BMI, hypertension and dyslipidemia. Smoking (defined by at 

least 100 smoked cigarettes and at least with a weekly smoking 

habit) was also associated with nfHCC (OR 2.18 adjusted 

for age and sex; 95% CI 1.23-3.83 p = 0.0067). In multivariate 

analysis, HBV infection (p=0.024), metabolic syndrome 

(P=0.0023) and tobacco use (p=0.0062) were independent


133 

Neurodevelopmental Outcomes in Patients with Biliary 

Atresia and Native Liver at Ages 1 and 2 Years: Results 

from ChiLDReN 

Vicky L. Ng 1 , Lisa G. Sorensen 16 , Estella M. Alonso 2 , Emily M. 

Fredericks 3 , Wen Ye 21 , Saul J. Karpen 17 , Benjamin Shneider 18 , 

Jorge A. Bezerra 4 , Jean P. Molleston 5 , Karen F. Murray 6 , Philip 

Rosenthal 7 , Kasper S. Wang 19 , Kathleen M. Loomes 8 , Paula M. 

Hertel 9 , Nanda Kerkar 20,22 , Kathleen B. Schwarz 10 , Yumirle P. 

Turmelle 11 , Barbara A. Haber 12 , Averell H. Sherker 15 , John C. 

Magee 13 , Ronald J. Sokol 14 ; 1 Division of Pediatric Gastroenterology, 

Hepatology and Nutrition, The Hospital for Sick Children, 

University of Toronto, Toronto, ON, Canada; 2 Division of Pediatric 

Gastroenterology, Hepatology and Nutrition, Ann & Robert H. 

Lurie Children’s Hospital, Chicago, IL; 3 Division of Child Behavioral 

Health, University of Michigan and C.S. Mott Children’s 

Hospital, Ann Arbor, MI; 4 Division of Pediatric Gastroenterology, 

Hepatology and Nutrition, Cincinnat Children’s Hospital Medical, 

Cincinnati, OH; 5 Pediatric Gastroenterology, Hepatology 

and Nutrition, Indiana Universtiy School of Medicine/Riley Hospital 

for Children, Indianapolis, IN; 6 Division of Gastroenterology 

and Hepatology, University of Washington Medical Center, 

Seattle Children’s, Seattle, WA; 7 Division of Gastroenterology, 

Hepatology and Nutrition, Department of Pediatrics, University 

of California, San Francisco, San Francisco, CA; 8 Pediatric Gastroenterology, 

Hepatology and Nutrition, Children’s Hospital of 

Philadelphia, Philadelphia, PA; 9 Baylor College of Medicine, 

Houston, TX; 10 Johns Hopkins School of Medicine, Baltimore, MD; 

11 Pediatric Gastroenterology, Hepatology and Nutrition, Washington 

University School of Medicine, St. Louis, MO; 12 Infectious 

Disease, Clinical Research, Merck, North Wales, PA; 13 University 

of Michigan Medical School, Ann Arbor, MI; 14 Section of Pediatric 

Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, 

University of Colorado School of Medicine, Denver, CO; 

15 Liver Diseases Research Branch, National Institute of Diabetes 

and Digestive and Kidney Diseases, National Institutes of Health, 

Bethesda, MD; 16 Ann & Robert H. Lurie Children’s Hospital of 

Chicago, Northwestern University Feinberg School of Medicine, 

Chicago, IL; 17 Pediatric Gastroenterology, Hepatology and Nutrition, 

Emory University School of Medicine/Children’s Healthcare 

of Atlanta, Atlanta, GA; 18 Pediatric Gastroenterology, Hepatology 

and Nutrition, Baylor College of Medicine, Houston, TX; 19 Division 

of Pediatric Surgery, Children’s Hospital Los Angeles, Los Angeles, 

CA; 20 Children’s Hospital of Los Angeles, Los Angeles, CA; 

21 Department of Biostatistics, University of Michigan, Ann Arbor, 

MI; 22 Mount Sinai, New York, NY 

Background: One of the most serious consequences of liver 

disease and malnutrition early in life is neurodevelopmental 

(ND) delay for which prompt identification and intervention 

may improve long-term outcomes. We hypothesized i) biliary 

atresia (BA) patients with their native liver at ages 1 and 2 

yrs will have significant ND delays and ii) that specific demographic 

and clinical variables will predict worse ND outcomes. 

Methods: Infants with BA, birth weight >2 kg, who received 

hepatoportoenterostomy (HPE) and were enrolled between 

2004-2012 in the multi-center, prospective NIDDK-funded 

ChiLDReN study (PROBE-NCT00061828) underwent ND testing 

at ages 12±2 mos (T1) and 24±2mos (T2) using either 

Bayley Scales of Infant Development-2(B2) or Bayley-3(B3), 

based on era. A subset of these infants was also enrolled in a 

blinded randomized placebo-controlled study of corticosteroids 

as adjunctive therapy to HPE (START–NCT00294684). Scores 

(normative mean=100±15) were categorized as ≥100, 85-99, 

70-84,


were included. Clinical and biochemical data were collected 

and compared with subsequent endoscopies to compare the 

efficacy and safety of prophylactic therapy versus non-treatment. 

Results: 273 endoscopies from 70 patients (42M) mean 

age 8.8 +/- 4.7y, were included. Diagnosis was portal vein 

thrombosis, biliary atresia, and other liver diseases, such as 

Wilson’s disease, autoimmune hepatitis, cystic fibrosis and 

congenital hepatic fibrosis in 23 (33%), 25 (35%) and 22 

(32%), respectively. Subjects were in the surveillance program 

for mean time of 4.8 years +/- 2.7 years (range, 9 months-8 

years). Variceal grading was as per UK national guidelines. 

Prophylactic treatment was administered in 177 (65%) endoscopies 

versus 96 (35%). Treatment group showed improvement 

of varices in 127 (72%) endoscopies, no change in 39 

(22%) and worsening of varices in 11 (6%). The non-treatment 

group showed improvement of varices in 16 (16%) endoscopies, 

no change in 38 (40%) and worsening of varices in 42 

(43%). Mean values for serum bilirubin, platelet count (PLT), 

haemoglobin (Hb) and INR at the time of endoscopy in the 

prophylaxis group and non-prophylaxis treatment group were 

18.65mg/dL and 21.11mg/dL [n.v. 3-20], 109.1x10 9 /L and 

103.95x10 9 /L [n.v. 180-400], 118.91g/L and 110.5g/L 

[n.v. 115-155], 1.28 and 1.25 [n.v. 0.9-1.2], respectively. 

No significant difference was found between treatment and 

non-treatment groups in bilirubin (p=0.4), PLT (p=0.4), Hb 

(p=0.38) or INR (p=0.5). Blood products prior to the procedure 

were required in 33 endoscopies (12%), as per protocol. 

Seven (10%) patients suffered a single GI bleed, all but one of 

which were in the no prophylactic treatment group. No other 

complications were recorded in either group. Conclusion: Prophylactic 

treatment of GI varices results in an overall reduction 

of varices and GI bleeds. Children with PTH could benefit from 

surveillance endoscopy programs with prophylactic treatment 

in the form of EVL or EST as it has been shown to be both safe 

and effective. 

Disclosures: 

The following authors have nothing to disclose: Harry Sutton, Mark Davenport, 

Alastair J. Baker, Anil Dhawan, Tassos Grammatikopoulos 

135 

Molecular genetic dissection of 101 neonatal/infantile 

intrahepatic cholestasis using targeted next-generation 

sequencing 

Takao Togawa, Tokio Sugiura, Koichi Ito, Takeshi Endo, Shinji 

Saitoh; Department of Pediatrics and Neonatology, Nagoya City 

University Graduate School of Medical Sciences, Nagoya, Japan 

Neonatal/infantile intrahepatic cholestasis (NIIC) is a heterogeneous 

disorder and caused by mutations in a number of 

genes, making molecular diagnosis challenging. Our aim is 

to clarify the molecular diagnosis for subjects with NIIC using 

next-generation sequencing (NGS) and bioinformatics pipeline. 

We further sought genotype-phenotype correlation to 

help comprehensive understanding of NIIC. Neonatal/infantile 

cholestasis was defined by the level of serum direct bilirubin; 

D.Bil≥1 mg/dL, if T.Bil8.6 kPa to detect METAVIR F3-F4 fibrosis. 

Several studies in children have used different LSM cut-points to 

identify fibrosis. However, validation of these LSM cut-points in 

children has not been reported. METHODS: Patients at Boston 

Children’s Hospital who underwent LSM from October 2006- 

May 2015 were eligible for this prospective two-phase study. 

All patients must have had a liver biopsy within 12 months 

prior to enrollment. The METAVIR system was used to stage 

fibrosis by pathologists blinded to LSM. Patients enrolled from 

October 2006- March 2011 were in the test group used to 

develop cut-points for discriminating F3-F4 and F4 fibrosis. 

Patients enrolled from April 2011-May 2015 were the validation 

group to test the accuracy of these cut-points. Diagnostic 

performance of TE was assessed by sensitivity (Se), specificity 

(Sp), accuracy (Ac) and area under the receiver operating 

characteristic (AUC). RESULTS: 263 subjects were enrolled; 97 

in the test group and 166 in the validation group. Clinical/ 

demographic data of the two groups were similar. Overall, 

the subjects were 46% female, aged 2 mo–24 yrs (median


13y), with indications for biopsy of autoimmune LD (19%), 

viral infection (18%), cholestatic LD (9%), PSC (12%), non-alcoholic 

fatty LD (11%), and post-transplantation (2%). METAVIR 

stages were: F0 (20%), F1 (29%), F2 (20%), F3 (17%), F4 

(14%). Median time from LSM to biopsy was 50 days (IQR 

21-144). Fasting prior to TE was known for 23%. In the test 

group, the cut-point to discriminate F3-F4 was >8.6 kPa with 

Se=79%, Sp=83%, Ac=81% and AUC=0.83. The cut-point for 

F4 was >11.5 kPa (Se=74%, Sp=82%, Ac=80%, AUC=0.81). 

Applied to the validation group, the cut-point of >8.6 kPa to 

detect F3-F4 had Se=67%, Sp=69% and Ac=69%. For subjects 

who fasted prior to TE (N=45), the performance for this 

cut-point improved (Se=92%, Sp=70%, Ac=76%). To detect 

F4, the cut-point of >11.5 kPa had Se=78%, Sp=80% and 

Ac=80%. In fasting subjects (N=45), the accuracy improved 

(Se=86%, Sp=84%, Ac=84%). CONCLUSIONS: This is the first 

study in children and young adults in which LSM cut-points for 

F3-F4 and F4 fibrosis are defined and evaluated in separate 

test and validation samples. This study supports recent findings 

that accuracy of TE improves with fasting. TE can be an important 

tool for identifying advanced fibrosis in children. 

Disclosures: 

Christine K. Lee - Grant/Research Support: Echosens 

Maureen M. Jonas - Advisory Committees or Review Panels: Gilead Sciences; 

Grant/Research Support: Gilead, Bristol Myers Squibb, Roche, Echosens 

The following authors have nothing to disclose: Paul D. Mitchell, Roshan Raza, 

Sarah Harney, Shanna M. Wiggins 

137 

Hospitalizations for Respiratory Syncytial Virus, Influenza, 

and Other Vaccine Preventable Illnesses in Liver 

Transplant Recipients at Freestanding US Children’s 

Hospitals 

Amy G. Feldman 1,4 , Brenda Beaty 2 , Shikha Sundaram 1,4 , Allison 

Kempe 3 ; 1 Pediatric Gastroenterology, Hepatology and Nutrition, 

Children’s Hospital Colorado, Aurora, CO; 2 University of Colorado, 

Aurora, CO; 3 Pediatrics, Children’s Hospital Colorado, 

Aurora, CO; 4 University of Colorado School of Medicine, Aurora, 

CO 

Objectives: Influenza and respiratory syncytial virus (RSV) are 

common infections requiring pediatric hospitalizations with 

annual rates of 0.05% and 0.5% respectively. Rates of these 

same illnesses in the pediatric liver transplant population are 

unknown. The goals of this study were to examine among liver 

transplant recipients at centers participating in the Pediatric 

Health Information System (PHIS) dataset: (1) number of hospitalizations 

for RSV, influenza, and other vaccine preventable 

illnesses (VPIs); (2) morbidity and mortality associated 

with these hospitalizations; and (3) costs associated with these 

hospitalizations. Methods: Retrospective cohort study of subjects 

< 18 years of age who underwent liver transplantation at 

a PHIS center between January 1, 2004, and December 31, 

2013. Subsequent hospitalizations during this time period for 

RSV, influenza, and 10 other VPIs were ascertained using ICD- 

9-CM diagnosis codes. Data were collected on clinical care, 

outcomes, and costs during these hospitalizations. Results: 

2,554 pediatric liver transplant recipients were identified; 

511 (20.0%) of the patients had a total of 799 cases of RSV, 

influenza, or other VPI. Overall, RSV, influenza, and rotavirus 

were the most common infections (7.2%, 6.6%, and 5.8% of 

cohort respectively). The mean time from transplant to infection 

was 1.7 years (SD 1.7 years). Ninety-two patients (3.6%) 

had infection during their transplant hospitalization, 8 of these 

had multiple infections. In the first year post-transplant, 4.0% 

of all patients had a hospitalization for RSV and 3.3% had 

a hospitalization for influenza. The death rate during these 

hospitalizations was 4.9% for RSV and 2.4% for influenza. 

Excluding those infections that occurred during initial transplant 

hospitalization (when all patients receive mechanical ventilation 

and ICU care), rejection occurred in 15.9% of patients 

with RSV and 13.4% of patients with influenza, ventilation in 

9.3% (RSV) and 5.9% (influenza), and ICU level care in 20.9% 

(RSV) and 12.8% (influenza). The median inflation adjusted 

costs for hospitalization were $ 15,934 (RSV) and $7806 

(influenza). Conclusions: VPIs occur in 1 of every 5 pediatric 

liver transplant recipients at a rate of up to 66 times higher than 

in the general pediatric population. RSV and influenza were 

the most common infections and most cases occurred in the 

first two years following transplant, demonstrating the importance 

of ensuring vaccination in all patients prior to liver transplant. 

Further research on clinical utility and cost-effectiveness 

of Palivizumab to prevent RSV in select transplant candidates/ 

recipients is needed. 

Disclosures: 

The following authors have nothing to disclose: Amy G. Feldman, Brenda Beaty, 

Shikha Sundaram, Allison Kempe 

138 

Liver Fibrosis is Present in One-Third of Adults with 

Alpha-1 Antitrypsin Deficiency Without Overt Liver Disease 

Virginia C. Clark 1 , George W. Marek 2 , Tracie L. Kurtz 3 , Chen 

Liu 4 , Farshid Rouhani 3 , Mark Brantly 3 ; 1 Medicine, Division of Gastroenterology, 

Hepatology, and Nutrition, University of Florida, 

Gainesville, FL; 2 Medicine, University of Florida, Gainesville, FL; 

3 Medicine, Pulmonary Critical Care, University of Florida, Gainesville, 

FL; 4 Pathology, University of Florida, Gainesville, FL 

Rationale: Alpha-1 antitrypsin deficiency (AATD) is a risk factor 

for the development of cirrhosis and liver related mortality in 

affected individuals. We hypothesize individuals with AATD 

have subclinical liver injury and fibrosis not detected by routine 

testing. The primary aim of this study is to define the prevalence 

and histologic spectrum of liver disease in adults with 

AATD. Methods: Adults with confirmed AATD Pi*ZZ and other 

rare AATD alleles participated in the IRB approved study after 

informed consent was obtained. A percutaneous liver biopsy 

was performed on eligible subjects. A single liver pathologist 

staged the biopsy for fibrosis (Ishak stage 1-6). The findings of 

AATD in PAS-D globules were scored qualitatively to describe 

the total observed (rare 0-1; moderate 2-3; severe 4-5). Individuals 

with a clinical history of decompensated liver disease were 

excluded. Results: The predominant AATD genotype is Pi*ZZ 

(n=73), and two patients with rare alleles are included Pi*ZM- 

Malton, 

and Pi*null. The mean age was 57 years, range 25-71 

and 61% (n=46) were female. The mean age of AATD diagnosis 

is 45.3 years, range 8 months-68 years. Pulmonary symptoms 

were present at AATD diagnosis in 64% (n=48) and 28% 

(n=21) were diagnosed because of family member. Only 5% 

(n=4) were identified by abnormal liver test. Pulmonary function 

characteristics included a baseline FEV1 1.95L (0.37-5.69L) 

and % predicted FEV1 of 61.5 (14-117). The prevalence of 

clinically significant liver fibrosis defined by Ishak fibrosis score 

≥2 is 34.7% (n=25). Men were more likely to have advanced 

fibrosis (Ishak ≥3, 22.2%, n=6) compared to women (2.2% 

n=1). ALT (22.5 vs 29.3 IU/L, p=0.03) and GGT (26.3 vs 

51.7, p=0.0006) were higher in those with Ishak ≥2, but mean 

values were within normal range. PAS-D globules correlated 

with fibrosis stage (spearman r= 0.5224, p


and risk for liver disease in AATD is needed to direct screening 

and novel therapies toward patients who could benefit most. 

In this cross section of older AATD subjects, over one-third had 

clinically significant liver fibrosis confirmed by biopsy. This is 

much higher than our previously reported prevalence of 7.9% 

using patient self-reported data. We also confirmed our previous 

finding that ALT and GGT for affected subjects fall within 

normal range, which limits clinical utility. The histologic spectrum 

was varied both in fibrosis and qualitative presence of 

AATD globules. Future studies will evaluate clinical risk factors 

and predictors for advanced liver disease. 

Disclosures: 

The following authors have nothing to disclose: Virginia C. Clark, George W. 

Marek, Tracie L. Kurtz, Chen Liu, Farshid Rouhani, Mark Brantly 

139 

Bile Acid Sequestrant Prevents NAFLD and NASH in 

Western Diet Fed Mice Independent of FXR 

Yuhuan Luo 1 , Xiaoxin Wang 1 , David J. Orlicky 2 , Moshe Levi 1 ; 

1 Medicine, Univ Colorado, Aurora, CO; 2 Pathology, University of 

Colorado, Aurora, CO 

Bile acid sequestrants (BAS) are orally administered nonabsorbable 

polymers that decrease serum cholesterol and serum glucose 

in patients and animal models of type 2 diabetes mellitus. 

We evaluated the effect of the BAS sevelamer in mice with diet 

induced obesity and insulin resistance that develop NAFLD and 

NASH. C57BL/6J male mice were fed with 1) control low fat 

(LF) diet or 2) western (WD) diet (high fat, high sucrose, cholesterol) 

for 5 months. The diets contained a) no addition or b) 2% 

sevelamer. Treatment with sevelamer prevented the increases 

in body weight, liver weight, serum triglycerides and serum 

cholesterol. Histopathological analysis revealed diet induced 

NAFLD and NASH mouse model resulted in panlobular macrosteatosis 

with some microvesicular steatosis, and increases in 

peri-portal inflammation and peri-portal and zone 1 sinusoidal 

fibrosis extending into zones 2 & 3 in places. Sevelamer treatment 

significantly decreased steatosis, inflammation and fibrosis 

scores. There was decreased FXR signaling in the intestine 

and the liver (FGF15 and SHP in the intestine as well as SHP 

in the liver). There was also increased liver bile acid synthesis 

(Cyp7a1) and expression of bile acid transporters. In addition, 

there were significant decreases in SREBP-1c, ACC and FAS, 

resulting in decreased neutral lipid accumulation. Sevelamer 

also prevented or reversed WD-induced upregulation of profibrotic 

(COL1A1, α-SMA, TGF-β) and proinflammatory (CCL2, 

IL-1β, and TNF-α) genes in the liver. These effects of sevelamer 

were independent of FXR as in FXR KO mice fed a western diet 

sevelamer had similar beneficial effects in improving NAFLD/ 

NASH as in wild type mice. BAS treatment therefore prevents 

diet induced NAFLD and NASH. This beneficial effect is largely 

independent of FXR but may be mediated through activation of 

the TGR5 signaling pathway. 

Disclosures: 

Moshe Levi - Grant/Research Support: Intercept, Genzyme-Sanofi, Daiichi Sankyo, 

Merck, Novartis 

The following authors have nothing to disclose: Yuhuan Luo, Xiaoxin Wang, 

David J. Orlicky 

140 

Targeting enterohepatic bile acid signaling as a novel 

approach to modulate hepatic autophagic activity in 

maintaining cholesterol homeostasis 

Yifeng Wang, Yifeng Ding, Hong-Min Ni, Wen-Xing Ding, Tiangang 

Li; Pharmacolgy, KUMC, Kansas City, KS 

Liver plays a key role in whole body cholesterol homeostasis. 

In addition, hepatic free cholesterol accumulation results in 

hepatocyte injury in non-alcoholic steatohepatitis (NASH). Lysosomes 

play an important role in maintaining cellular cholesterol 

homeostasis by regulating cholesterol ester hydrolysis that 

affects pathways such as cholesterol uptake, secretion and bile 

acid synthesis. New studies revealed that autophagy mediates 

the cellular transport of cholesterol ester from lipid droplets to 

lysosomes. Goal: this study investigated the role and regulation 

of hepatic autophagy in cholesterol metabolism. Methods: the 

effect of cholesterol accumulation on autophagy flux and lysosome 

function was investigated in hepatocytes and in mice. 

The impact of autophagy impairment on hepatic and plasma 

cholesterol metabolism was studied in liver-specific ATG5 

knockout mice. The effect of cholestyramine on hepatic autophagy 

was studied in mice. Results: Feeding mice a cholesterol-containing 

diet or an atherogenic diet for 6 days increased 

liver LC3-II and p62 levels in response to hepatic cholesterol 

accumulation. Both free cholesterol loading and treatment with 

low density lipoprotein (LDL) increased LC3B-II and p62 levels 

in human hepatocytes and HepG2 cells, however, only free 

cholesterol loading, but not LDL treatment, increased lysosome 

size and decreased cathepsin B activities in HepG2 cells, suggesting 

free cholesterol accumulation can also impair lysosome 

function. Consistently, inhibition of cholesterol esterification 

with an ACAT inhibitor to increase cellular free cholesterol 

levels blocked autophagosome/lysosome fusion and caused 

CASPASE-3 activation in HepG2 cells. In autophagy-defective 

liver-specific ATG5 knockout mice, hepatic cholesterol was 

unchanged, but FPLC analysis showed that plasma LDL-cholesterol 

was significantly elevated. As previous studies suggested 

that autophagy was impaired in NASH, we found that cholestyramine 

feeding increased hepatic autophagy in both lean 

and ob/ob mice, which was possibly due to the removal of 

bile acid-mediated inhibition of autophagosome/lysosome 

fusion. Conclusion: this study suggests that hepatic autophagy 

is required for maintaining whole body cholesterol homeostasis. 

Hepatic free cholesterol accumulation impairs hepatic 

autophagy, which exacerbates hypercholesterolemia and 

hepatocyte-injury in NASH. As we recently showed that bile 

acids inhibit hepatic autophagy, targeting enterohepatic bile 

acid signaling by bile acid binding resin may be an effective 

approach to induce hepatic autophagy and attenuate cholesterol-induced 

liver injury. 

Disclosures: 

The following authors have nothing to disclose: Yifeng Wang, Yifeng Ding, Hong- 

Min Ni, Wen-Xing Ding, Tiangang Li 

141 

Anti-Fibrotic Effect of Autotaxin and LPA1R Antagonists 

in a Rodent Model of NASH. 

Malavi Madireddi 1 , Gretchen Bain 3 , James Swaney 2 , Brian J. 

Murphy 1 , Simeon Taylor 1 ; 1 Metabolic and Fibrotic Diseases, Bristol 

Myers Squibb, West Windsor, NJ; 2 Inception Sciences, San 

Diego, CA; 3 PharmAkea, Inc., San Diego, CA 

Nonalcoholic steatohepatitis (NASH) is a condition where 

increased fat accumulation concomitant with inflammation 

results in liver cell damage. Patients with NASH have elevated


serum lipids and diabetes. Early disease is clinically asymptomatic, 

therefore often presenting only when critical hepatic 

function is compromised. A wound healing response in general 

occurs in injured liver tissue, and the persistence of this 

response may result in liver fibrosis. At this time, there is no 

treatment for NASH, and left untreated leads to hepatocellular 

carcimona and liver cirrhosis. Current management focuses on 

treating underlying cause of injury, as there are no targeted 

therapeutics to interrupt fibrotic response; liver transplant is 

the only option for late-stage cirrhosis. The enzyme Autotaxin 

(ATX) hydrolyzes lysophosphatidylcholine to produce lysophosphatidic 

acid (LPA), a multi-functional bioactive lipid mediator. 

ATX is a major determinant of LPA levels in circulation, and the 

pathophysiological function of ATX is attributed to its ability 

to produce LPA. There is mounting literature supporting serum 

ATX as an indicator of the severity of liver disease. Here we 

investigated two small molecule inhibitors of ATX and LPA1 

receptor in the STAM ® rodent model of liver fibrosis to pharmacologically 

interrogate these targets in NASH. AM063 and 

BMT-053011 are low nanomolar inhibitor of ATX and LPA1 

receptor respectively. The STAM ® mouse model of NASH is 

induced by a single subcutaneous injection of streptozotocin 

two days after birth and by feeding a high fat diet. Starting at 

6 weeks after disease initiation, BMT-053011 was administered 

twice daily, while AM063 and Telmisartan were administered 

once daily. Treatment was continued for the following 3 

weeks after initiation. AM063 showed significant decreases in 

plasma TG, fibrosis area and TNF-a expression levels, with a 

decreasing trend in blood glucose and macrophage activation 

compared to the Vehicle group. Treatment with BMT-053011 

showed significant decreases in liver weight, NAS, the fibrosis 

area and inflammation. The results, together with the decrease 

in ALT and inflammatory gene expression levels, demonstrate 

anti-inflammatory and hepatoprotective effects. Importantly, as 

NAS is one of the clinical endpoints for assessing the activity 

of NASH (Sanyal AJ. et al., Hepatology, 2011; 54:344), the 

observed changes in the treatment groups suggest potential 

translatability of BMT-053011 as an anti-NASH therapeutic 

for humans. 

Disclosures: 

Malavi Madireddi - Employment: Bristol Myers Squibb 

Gretchen Bain - Employment: PharmAkea Therapeutics 

Simeon Taylor - Consulting: Isis Pharmaceuticals, Aegerion, Yabao; Stock Shareholder: 

Bristol-Myers Squibb, Gilead, Celgene 

The following authors have nothing to disclose: James Swaney, Brian J. Murphy 

142 

Treatment with the FXR-TGR5 dual agonist INT-767 

arrests and reverses progression of NASH in mice fed a 

Western diet 

Xiaoxin Wang 1 , Yuhuan Luo 1 , David J. Orlicky 2 , Luciano Adorini 3 , 

Moshe Levi 1 ; 1 Medicine, Univ Colorado, Aurora, CO; 2 Pathology, 

University of Colorado, Aurora, CO; 3 Intercept Pharmaceuticals, 

New York, NY 

Obesity and insulin resistance have become leading causes 

of nonalcoholic fatty liver disease (NAFLD) and nonalcoholic 

steatohepatitis (NASH). In the present study, we determined in 

mice with established NASH if treatment with INT-767, a dual 

agonist of the nuclear receptor farnesoid X receptor (FXR) and 

the G protein coupled receptor TGR5, can decrease disease 

progression. In these studies, C57BL/6Jmice were first fed a 

low fat (LF) or a Western diet (WD: high fat, high sucrose, and 

high cholesterol) for 3 months and then treated orally for an 

additional 2 months with vehicle or INT-767, 30 mg/kg body 

weight/d. Compared to mice fed a LF diet, feeding a WD 

for 5 months resulted in panlobular macrosteatosis with some 

microvesicular steatosis, increased peri-portal inflammation and 

lipogranuloma formation, and peri-portal and zone 1 sinusoidal 

fibrosis extending in places into zones 2 & 3. When mice 

on WD were treated with INT-767, there were marked and significant 

decreases in the percentage of mice showing hepatic 

macrosteatosis (83%), microsteatosis (100%), inflammation 

(100%) and fibrosis (47%) compared to non-treated WD fed 

mice. These histopathological changes were accompanied by 

significant decreases in liver cholesterol content (52%), as well 

as decreases in the lipogenic transcription factor SREBP-1c, 

pro-inflammatory mediator MCP-1, and profibrotic matrix protein 

Col1a1. INT-767 treatment also increased liver expression 

of proteins involved in mitochondrial biogenesis and energy 

expenditure including AMPK, Sirtuin 1, PGC-1α, Sirtuin 3, and 

ERRα. Using FXR knockout mice, the FXR selective agonist INT- 

747 (obeticholic acid), and the TGR5 specific agonist INT-777, 

we further determined that FXR plays a major role in mediating 

the effects of INT-767 in the liver, including the increases in 

PGC-1α and Sirtuin 3. The dual FXR-TGR5 agonist INT-767 is 

therefore able to markedly and significantly arrest and reverse 

progression of liver disease even when treatment is started in 

the presence of obesity, insulin resistance, and NASH. 

Disclosures: 

Luciano Adorini - Employment: Intercept Pharmaceuticals 



The following authors have nothing to disclose: Xiaoxin Wang, Yuhuan Luo, 


143 

DRX-065, the stabilized R-enantiomer of pioglitazone is 

without PPARg agonist activity and exhibits the beneficial 

in vivo pharmacodynamic effects for the treatment 

of NASH 

Sheila H. DeWitt, Vincent Jacques, Lex H. Van der Ploeg; DeuteRx, 

Andover, MA 

Background: Pioglitazone is approved for the treatment of 

type 2 diabetes mellitus (T2DM) and is currently the only drug 

recommended off-label to treat patients with biopsy-proven 

nonalcoholic steatohepatitis (NASH). However, the use of 

pioglitazone for NASH has been limited due the side effect 

of weight gain. Pioglitazone is a mixture of two mirror-image 

compounds (the R- and S-enantiomers) that rapidly interconvert 

in vivo. The pharmacokinetic (PK) and pharmacodynamic 

(PD) properties of the individual enantiomers has never been 

reported in humans. Methods: We replaced the exchangeable 

hydrogen at the chiral center of pioglitazone by deuterium, 

a stable, naturally occurring isotope of hydrogen. Deuterium 

at the chiral center stabilizes the enantiomers and reduces 

their rate of interconversion. Stability studies were completed 

in aqueous buffer and in mouse and human plasma. In vitro 

PPARg agonist activity was determined in a TRAP220 coactivator 

recruitment assay. In vitro mitochondrial respiration activity 

was assessed in intact C2C12 cells. PK and weight gain studies 

of the enantiomers were completed in male C57BL/6 mice. 

Efficacy was evaluated in a db/db mouse model of T2DM, 

where effects on metabolic and inflammatory markers were 

assessed. Results: Preparation of the deuterium-stabilized enantiomers 

of pioglitazone, d-R-pioglitazone (DRX-065) and d-S-pioglitazone 

(d-S-pio) enabled the evaluation of the properties of 

the individual enantiomers. DRX-065 is not a PPARg agonist 

at concentrations up to 100 mM while d-S-pio accounts for 

all of the PPARg agonist activity observed with racemic pioglitazone. 

DRX-065 is exclusively responsible for the effects of


pioglitazone on mitochondrial respiration. In the db/db mouse 

model of T2DM, DRX-065 significantly decreases blood glucose, 

serum triglycerides, and fatty acids, as well as markers of 

inflammation (cytokines, serum amyloid A). At a therapeutically 

effective dose in mice, DRX-065 also does not induce weight 

gain, unlike pioglitazone or d-S-pio. The effects of d-S-pio on 

weight gain are expected based on its strong PPARg agonist 

activity. Conclusion: We demonstrated in preclinical experiments 

that the stabilized deuterated R-enantiomer of pioglitazone, 

DRX-065, has pharmacological properties desirable 

for the treatment of NASH (mitochondrial function modulation, 

non-steroidal anti-inflammatory effects, and glucose lowering 

effects) without the undesired PPARg-related weight gain side 

effects. DRX-065 represents a potentially significant therapeutic 

improvement over pioglitazone, a drug already recommended 

off-label for the treatment of NASH. 

Disclosures: 

The following authors have nothing to disclose: Sheila H. DeWitt, Vincent 

Jacques, Lex H. Van der Ploeg 

144 

Peretinoin, an acyclic retinoid, suppresses steatohepatitis 

and development of tumorigenesis by activated 

Atg16L1-dependent autophagy in mice fed an atherogenic 

high-fat diet. 

Hikari Okada 1 , Masao Honda 1 , Kai Takegoshi 1 , Taro Yamashita 1 , 

Naoto Nagata 2 , Toshinari Takamura 2 , Takuji Tanaka 3 , Shuichi 

Kaneko 1 ; 1 Gastroenterology, Kanazawa University Graduate 

School of Medical Science, Kanazawa, Japan; 2 Department of 

Disease Control and Homeostasis, Kanazawa University Graduate 

School of Medical Sciences, Kanazawa, Japan; 3 Cancer Research 

and Prevention, The Tohkai Cytopathology Institute, Gifu, Japan 

Objective Peretinoin, acyclic retinoid, is under phase III clinical 

trials in Japan to prevent the recurrence and development of 

HCC after surgical removal of the primary tumors. The mechanism 

of peretinoin for preventing HCC remains unclear. Mice 

fed an atherogenic high-fat diet (Ath HFD) developed steatohepatitis 

followed by hepatic fibrosis and HCC progression 

(Hepatology 2007). Here we investigated the suppressive 

effects of peretinoin on steatohepatitis and tumorigenesis in 

Ath HFD mice. Materials and Methods Three groups of 8-weekold 

mice (n=15-20/group) were fed a control diet or Ath HFD 

containing 0.01% or 0.03% peretinoin for 12, 30, and 60 

weeks. Then, 0.01% peretinoin was added to the Ath HFD 

at 40 weeks to examine the reversible effect of peretinoin on 

established fibrosis and steatosis in the liver. The degree of liver 

steatosis, hepatic fibrosis, tumor incidence, and liver weight 

was calculated. Expression of IL6, IL1β TNFα, CEBPα, collagen 

I/IV, pSTAT3, pNFkβ, ATG5, ATG7, ATG16L1, LC3b, 

and Lamp2 was evaluated by immunohistochemical staining, 

real-time PCR, and western blotting. The promoter analysis of 

ATG16L1 was performed by reporter gene assay and ChIP 

assay. Primary hepatocytes were isolated from male C57BL/6 

mice treated Ath HFD for 12 weeks and the effect of peretinoin 

on primary hepatocytes was evaluated in vitro. Results Mice 

fed an Ath HFD developed liver steatosis and liver fibrosis after 

12 and 30 weeks, and developed liver tumors after 60 weeks 

at 70% frequency. Peretinoin markedly reduced steatosis and 

fibrosis at 12 and 30 weeks and reduced tumor incidence to 

approximately 30%. Expression of IL6, IL1β, TNFα, collagen 

I/IV, pSTAT3, and pNFkβ was suppressed to approximately 

60% in mice fed an Ath HFD containing peretinoin compared 

with mice fed only an Ath HFD. We found peretinoin induced 

autophagy that was shown by the increased autophagosome 

formation by electron microscopy. We found increased LC3-II 

and co-localization of autophagosome (LC3-II) and lysosome 

(Lamp2) by immune fluorescent staining. Among various autophagy 

related factors, peretinoin increased the expression of 

Atg16L1 and promoted Atg16L1-dependent autophagy. In primary 

hepatocyte, palmitic acid and IL6 suppressed the expression 

of Atg16L1, while peretinoin rescued the Atg16L1 and 

suppressed pSTAT3 and pNFkβ. We found peretinoin induced 

the expression of CEBPα and CEBPα activated the promoter 

activity of Atg16L1. Conclusion Peretinoin suppresses the development 

of liver steatosis, inflammation, and tumorigenesis by 

activating Atg16L1-dependent autophagy. These results support 

the clinical efficacy of peretinoin for preventing HCC. 

Disclosures: 

Hikari Okada - Employment: Kanazawa University 

Shuichi Kaneko - Grant/Research Support: MDS, Co., Inc, Chugai Pharma., Co., 

Inc, Toray Co., Inc, Daiichi Sankyo., Co., Inc, Dainippon Sumitomo, Co., Inc, 

Ajinomoto Co., Inc, Bristol Myers Squibb., Inc, Pfizer., Co., Inc, Astellas., Inc, 

Takeda., Co., Inc, Otsuka„ÄÄPharmaceutical, Co., Inc, Eizai Co., Inc, Bayer 

Japan, Eli lilly Japan 

The following authors have nothing to disclose: Masao Honda, Kai Takegoshi, 

Taro Yamashita, Naoto Nagata, Toshinari Takamura, Takuji Tanaka 

145 

Effects of norfloxacin therapy on survival in patients 

with Child-Pugh class C cirrhosis: Results of a randomized, 

double-blind, placebo-controlled, multicenter trial 

Richard Moreau 1,2 , Laure Elkrief 2 , Christophe Bureau 3 , Jean- 

Marc Perarnau 4 , Thierry Thevenot 5 , Faouzi Saliba 6 , Isabelle Ollivier-Hourmand 

15 , Alexandre Louvet 7 , Pierre Nahon 8 , Frédéric 

Oberti 9 , Rodolphe Anty 10 , Sophie Hillaire 11 , Blandine Pasquet 12 , 

Violaine Ozenne 13 , Marika Rudler 14 , Marie Angele Robic 3 , Louis 

d’Alteroche 4 , Vincent Di Martino 5 , Pierre-Emmanuel Rautou 2 , Nathalie 

Gault 16 , Didier Lebrec 1,2 ; 1 UMR S1149, Center for Research 

on Inflammation (CRI), Inserm and Paris Diderot University, Clichy, 

France; 2 DHU UNITY, Service d’hépatologie, Hôpital Beaujon, 

APHP, Clichy, France; 3 Service d’hépato-gastroentérologie,, Hôpital 

Purpan CHU Toulouse et Université Paul Sabatier, Toulouse, 

France; 4 Unité d’Hépatologie, Hépatogastroentérologie, CHRU 

Tours, Tours, France; 5 Service d’Hépatologie et de Soins Intensifs 

Digestifs, Hôpital Jean Minjoz, Besançon, France; 6 Centre 

Hépato-Biliaire, Hôpital Paul Brousse, APHP, Villejuif, France; 

7 Service des maladies de l’appareil digestif, Hôpital Huriez, Lille, 

France; 8 Service d’Hépatologie, Hôpital Jean Verdier, APHP, 

Bondy, France; 9 Service d’hépato-gastroentérologie, CHU d’Angers, 

Angers, France; 10 Pôle Digestif and Inserm 1065, CHU 

de Nice, Nice, France; 11 Service de Médecine Interne, Hôpital 

Foch, Suresnes, France; 12 Unité de recherche clinique Paris Nord, 

Hôpital Bichat, APHP, Paris, France; 13 Hépato-gastroentérologie, 

Hôpital Lariboisière, APHP, Paris, France; 14 Hépatologie, GH 

Pitié-Salpêtrière, APHP, Paris, France; 15 Service d’hépato-gastro-entérologie 

et nutrition and Unité 1930, CHU Côte de Nacre, 

Caen, France; 16 Unité de recherche clinique Paris Nord, Hôpital 

Beaujon, APHP, Clichy, France 

Background & Aims: Whether long-term oral antibiotic therapy 

using a fluoroquinolone should be used to improve prognosis in 

patients with advanced cirrhosis is debated. Thus we addressed 

this question by investigating the effects of the administration 

of norfloxacin (a fluoroquinolone) in patients with advanced 

cirrhosis. Methods: We performed a multicenter, double-blind 

trial in which we randomly assigned patients with Child-Pugh 

class C cirrhosis to once-daily 400 mg oral norfloxacin (144 

patients) or placebo (147 patients) for 6 months. Surviving 

patients were followed-up for further 6 months after treatment 

cessation. The primary end point was overall survival at 6 

months; secondary end points included overall survival at 3 

and 12 months, and liver-related complications (bacterial infec-


tion, kidney failure, encephalopathy, variceal hemorrhage). 

Patients were censored at the time of liver transplantation or 

spontaneous bacterial peritonitis. Results: 291 patients were 

enrolled rather than the projected 392 because of slow recruitment 

and termination of funding. Excessive alcohol consumption 

was the cause of cirrhosis in 115 patients (79.9%) of the 

norfloxacin group and 108 patients (73.5%) of the placebo 

group. By 6 months, 19 deaths (13.2%) occurred in the norfloxacin 

group as compared with 27 deaths (18.4%) in the 

placebo group, with a hazard ratio (HR) for death at 6 months 

of 0.69 (95% confidence interval [CI], 0.38 to 1.24; P=0.21). 

The HR for death at 3 months and 12 months were 0.53 (95% 

CI, 0.28 to 1.02; P=0.05) and 0.76 (0.48 to 1.22; P=0.26), 

respectively. There were no significant differences in terms of 

6-month rates of infections (20.8% vs. 22.4%), kidney failure 

(6.9% vs. 7.5%), encephalopathy (16.7% vs. 19.0%), variceal 

hemorrhage (3.5% vs. 4.1%). Post-hoc analysis showed that at 

enrollment, there were 56 patients (39.2%) in the norfloxacin 

group and 63 (42.9%) in the placebo group who had histological 

features of alcoholic hepatitis (P=0.52). In this subgroup 

of patients, the HR for death at 3 months, 6 months and 12 

months were 0.32 (95% CI, 0.12 to 0.86; P=0.01), 0.49 

(95% CI, 0.21 to 1.15; P=0.09) and 0.47 (95% CI, 0.23 to 

0.97; P=0.03), respectively. Conclusions: In this study, 6-month 

oral norfloxacin therapy did not improve overall survival and 

did not affect the development of liver-related complications 

in patients with Child-Pugh class C cirrhosis. In the subgroup 

of patients with alcoholic hepatitis, norfloxacin therapy may 

increase 3-month overall survival suggesting early beneficial 

effects of the antibiotic in this subgroup of patients. 

Disclosures: 

Christophe Bureau - Speaking and Teaching: Gore 

Jean-Marc Perarnau - Consulting: Gore and Associates 

Faouzi Saliba - Advisory Committees or Review Panels: Novartis, Roche, Genzyme, 

Vital therapies; Grant/Research Support: Astellas; Speaking and Teaching: 

Gambro, MSD, Gilead 

Isabelle Ollivier-Hourmand - Speaking and Teaching: gilead, jansen, bayer 




Pierre-Emmanuel Rautou - Speaking and Teaching: Gilead 

The following authors have nothing to disclose: Richard Moreau, Laure Elkrief, 

Thierry Thevenot, Alexandre Louvet, Pierre Nahon, Frédéric Oberti, Rodolphe 

Anty, Sophie Hillaire, Blandine Pasquet, Violaine Ozenne, Marika Rudler, Marie 

Angele Robic, Louis d’Alteroche, Nathalie Gault, Didier Lebrec 

146 

Ischemic hepatitis following acute variceal bleed is 

ameliorated by N- acetyl cysteine (NAC) in cirrhotics: A 

prospective randomized controlled trial 

Avinash Kumar, Ankur Jindal, Rakhi Maiwall, Lovkesh Anand, Amrish 

Sahney, Shiv K. Sarin; Institute Of Liver and Biliary Sciences, 

New Delhi, India 

Background & aims. Ischemic hepatitis (IH) following acute 

variceal bleed (AVB) in cirrhotics carries ominous prognosis. 

Consequent hypotension & hepatic ischemia may result in centrilobular 

necrosis . N-acetylcysteine (NAC), a potent anti-oxidant 

may prevent cell injury & improve tissue oxygen delivery. 

We investigated the efficacy & safety of NAC in the prevention 

of IH & survival. Methods: Of 530 consecutive patients 

admitted with UGI bleed (Aug 2013-Apr 2015), cirrhotics with 

acute variceal bleed (n=214) were randomized in a prospective 

open label study (NCT02015403) to receive either standard 

of care (SOC; iv fluids, terlipressin, endoscopic variceal 

ligation, blood/blood products & antibiotics) plus NAC (150 

mg/kg/hr for 1 h followed by 12.5mg/kg/hr for 4 h, followed 

by 6.25 mg/kg for 67 hrs) [Group A(GrA), n=107) or 

SOC alone [Group B(GrB),n=107]. Patients were monitored 

for presence & severity of IH (increase in AST of ≥5 X ULN & 

bilirubin ≥1.5 X ULN in


patients, 50(4.9%) were found to have definite HP; 4 were 

excluded (2 on L-dopa and 2 on bromocriptine).The patients 

in Gr A and B were comparable in age (54.8± 6.7 vs., 54.3 

±7.4 yr, p=0.7) gender(Males 86%vs. 83%, p= 0.5),etiology 

of cirrhosis, MELD (17.9 ± 3.8vs. 18.2 ± 4.2,p=0.3), 

baseline arterial ammonia(131 ± 7.4vs. 132 ± 7.9mmol/l, 

p=.4), number of patients with recurrent HE: 13/22(59%) 

vs. 14/24(58%), p=0.7,persistent HE: 8/22 (36%) vs. 9/24 

(37.5%), p=0.6, and porto-systemic shunts: 17/21(80) vs. 

19/24(79), p= 0.6. Complete &partial response was seen in 

(none vs. 7 (29%), p


on the underlying mechanisms of action of taurine and further 

long-term evaluation in cirrhosis and portal hypertension are 

warranted. 

Disclosures: 

Mattias Mandorfer - Consulting: Janssen; Speaking and Teaching: AbbVie, Gilead, 

Janssen, Boehringer Ingelheim, Bristol-Myers Squibb, Roche 

Thomas Reiberger - Consulting: Xtuit; Grant/Research Support: Roche, Gilead, 

MSD, Phenex; Speaking and Teaching: Roche, Gilead, MSD 









The following authors have nothing to disclose: Remy Schwarzer, Rafael Paternostro, 

Birgit B. Heinisch, Philipp Schwabl, Arnulf Ferlitsch 

150 

Anticoagulation in patients with cirrhosis and portal 

vein thrombosis is associated with increased portal vein 

recanalization and better prognosis. 

Carlos Noronha Ferreira 1 , Teresa Rodrigues 2 , Ana Júlia Pedro 3 , 

Paula Ferreira 1 , Margarida Sobral Dias 1 , Afonso Gonçalves 4 , 

Leonor Xavier Brito 1 , Fatima Serejo 1 , Rui T. Marinho 1 , Cilénia 

B. Costa 1 , Narcisa Fatela 1 , Helena Cortez-Pinto 1 , Fernando 

Ramalho 1 , Paula Alexandrino 1 , José F. Velosa 1 ; 1 Serviço de Gastrenterologia 

e Hepatologia, Hospital de Santa Maria, Centro 

Hospitalar Lisboa Norte, Lisboa, Portugal; 2 Laboratório de Biomatemática, 

Faculdade de Medicina de Lisboa, Lisboa, Portugal; 

3 Serviço de Medicina 2, Hospital de Santa Maria - Centro Hospitalar 

Lisboa Norte, Lisboa, Portugal; 4 Serviço de Imagiologia, 

Hospital de Santa Maria, Centro Hospitalar Lisboa Norte, Lisboa, 

Portugal 

Introduction: Cirrhosis is recognized as a prothrombotic state. 

A recent study showed that prophylactic anticoagulation prevented 

portal vein thrombosis (PVT) and decreased episodes 

of decompensation of cirrhosis. Aims: To analyze the effect 

of anticoagulation on recanalization of non-tumoral PVT in 

patients with cirrhosis and its effect on prognosis. Methods: 69 

consecutive patients with cirrhosis diagnosed with non-tumoral 

PVT were studied. The clinical features at diagnosis of PVT and 

factors associated with anticoagulation use were studied. Decision 

to start anticoagulation was taken at the discretion of the 

clinician managing the patient. The effect of anticoagulation 

on PVT recanalization and mortality was analyzed. Results: 

The average age was 58.6±11.8 years and 44(64%) were 

males. Severity of cirrhosis: Median(Range) Child–Pugh(CP) 

score: 8(5-15), MELD score:13(6-35). CP class: A-15(22%), 

B-32(46%), C-22(32%). At diagnosis of PVT, 55(80%) were 

symptomatic. Variceal bleeding(VB) in 30(46%) and abdominal 

pain in 19(29%) were the main clinical presentations. 

Anticoagulation (LMWH–9, warfarin–16) was administered in 

25(36%) patients one of whom with cavernoma. Patients with 

VB were less likely to be given anticoagulation (p=0.037). 

There were no differences in age, gender, etiology, severity of 

cirrhosis and extent of PVT in patients receiving, or not, anticoagulation. 

Recanalization of PVT was assessed by at least one 

imaging study in 60 patients and recanalization (Total–13, partial 

– 9) of the portal vein was documented in 22(37%) patients. 

Median (Range) follow–up was 21(0-376) months. At the end 

of follow-up, 29(42%) patients died, of which sixteen deaths 

were related to infectious complications with no deaths due to 

anticoagulation related bleeding. By Cox regression analysis, 

factors associated with mortality at the end of follow-up were: 

Age (HR 1.040, 95% C.I. 1.002–1.078, p=0.037), CP score 

(HR 1.35, 95% C.I. 1.18–1.55, p


Additional resources from government for HCV could aid in the 

elimination of HCV from the United States. 

Cost of HCV in the era of DAAs 

($1.5 billion USD), shielding x-ray components ($13.2 billion 

USD), and reducing greenhouse gas emissions ($413 billion 

USD). Conclusions: The scope of benefit conferred by new 

HCV therapies is large due to the high mortality risk of HCV, 

the curative effect of therapy and the size of the population 

infected. However, at current prices the incremental cost of 

therapy represents a 3478-fold increase in current nationwide 

spending on hepatitis C, more than 11% of all national health 

expenditures, and 11 times the annual NIH budget. Concurrent 

efforts to increase access to treatment as well as reduce drug 

costs should be prioritized. 

Disclosures: 

The following authors have nothing to disclose: Mai T. Pho, Andrew I. Aronsohn, 

Benjamin P. Linas, Daniel C. Johnson, Jake R. Morgan, William V. Padula, David 

O. Meltzer 

Disclosures: 

Jagpreet Chhatwal - Consulting: Merck & Co., Inc., Gilead, Complete HEOR 

Solutions; Grant/Research Support: NIH/National Center for Advancing Translational 

Sciences 

The following authors have nothing to disclose: Xiaojie Wang, Mark S. Roberts, 

Mina Kabiri, Turgay Ayer, Julie M. Donohue, Fasiha Kanwal 

152 

Treating Hepatitis C in the US: measuring impact and 

value in the context of other major health interventions 

Mai T. Pho 1 , Andrew I. Aronsohn 1 , Benjamin P. Linas 2 , Daniel 

C. Johnson 1 , Jake R. Morgan 2 , William V. Padula 1 , David O. 

Meltzer 1 ; 1 Medicine, University of Chicago, Chicago, IL; 2 Boston 

Medical Center, Boston, MA 

Background: Hepatitis C (HCV) results in significant morbidity 

and mortality in the US. All-oral therapies, while curative, are 

costly. We estimate the economic value of providing treatment 

for individuals with HCV relative to other major interventions 

such as decreasing greenhouse gas emissions. Methods: We 

calculate the net monetary benefit (NMB) of therapy provided 

for all individuals estimated to be infected with HCV in the US 

using a value of statistical life (VSL) and value of statistical life 

year (VSLY) framework. A VSL of 5 million US dollars (USD) is 

used, and we calculate the VSLY as the quotient of the VSL and 

the discounted expected remaining life-years for the general 

population. We estimate the total population of HCV based 

on prevalence, demographic and genotype data from the 

National Health and Nutrition Evaluation Survey from 2003- 

2010. The NMB of therapy is the product of VSLY and the 

remaining discounted life-years associated with treatment and 

no treatment of the total population, subtracted by the incremental 

cost. Regimens examined included 12 weeks of sofosbuvir 

(SOF) and ledipasvir (LDV) for genotype (GT)1, 12-16 

weeks of SOF and ribavirin (RBV) for GT2, and 24 weeks of 

SOF and RBV for GT3 infections. Remaining life expectancy 

and costs are projected using the Hepatitis C Virus Cost-Effectiveness 

(HEP-CE) simulation model. Prices for HCV drugs are 

varied from 100% to 55% to reflect offsets due to rebates and 

other discounts. Benefits and costs are reported in the context 

of federally regulated health interventions. Results: Compared 

to no treatment, treating 2.7 million individuals with HCV will 

result in 9.5 million life-years gained at an incremental cost of 

$347 billion USD. The NMB of HCV therapy is $2.2 trillion 

USD. When the price of medications is discounted by 45%, 

the NMB of therapy is $2.4 trillion USD, with an incremental 

cost of therapy of $187 billion USD. The NMB of HCV 

treatment exceeds influenza vaccinations at long-term facilities 

153 

Hepatitis C (HCV) Infection in Baby Boomers Are Independently 

Associated with Mortality and Resource Utilization 

Mehmet Sayiner 2 , Mark Wymer 2 , Pegah Golabi 2 , Joel S. Ford 3 , 

Indie Srishord 2 , Zobair M. Younossi 2,1 ; 1 Center For Liver Disease, 

Department of Medicine, Inova Fairfax Hospital, Falls Church, 

VA; 2 Betty and Guy Beatty Center for Integrated Research, Inova 

Health System, Falls Church, VA; 3 Department of Medicine, Inova 

Health System, Falls Church, VA 

Background and aim: HCV is more common among Baby 

boomers (BB) (born 1946-1965). As this cohort age, they will 

increasingly become Medicare eligible. Our aim was to evaluate 

resource utilization and short term mortality of BB-Medicare 

recipients with HCV. Method: We used inpatient and outpatient 

Medicare databases (2005-2010) with clinical data, 

admission and discharge information, ICD-9 codes, procedure 

codes, and billing information. Medicare patients who were 

considered BB were identified using all HCV-related ICD-9 

codes. Charlson Index was calculated as a measure of comorbidity. 

The outcome measures included resource utilizations 

[payment per case and inpatient length of stay (LOS)] and short 

term mortality (90 days). Results: We included 1,153,862 

BB who received Medicare services from 2005 to 2010. Of 

these, 3.2% (N=37,365) were HCV (+). During this period, 

Medicare-BB in the inpatient setting increased from 39,793 to 

55,235, and their claims increased from 78,924 to 106,232. 

During the study period, both overall mortality [8.94% to 

10.25% (p


category of 45-49 (4.21%, [3.14-5.28]), ESRD (966.31%, 

[954.86-977.88]), disabled status (43.22%, [41.67-44.80]), 

Charlson score (46.78%, [46.31-47.26]), and year study 

(2.72%, (2.58-2.85]) were all independently associated with 

an increase in total payments. Conclusions: Prevalence of HCV 

is higher in Baby boomers Medicare recipients. For the entire 

BB cohort, HCV positivity is independently associated with 

higher mortality and resource utilization. 

Disclosures: 



The following authors have nothing to disclose: Mehmet Sayiner, Mark Wymer, 

Pegah Golabi, Joel S. Ford, Indie Srishord 

154 

Bariatric surgery for the treatment of nonalcoholic steatohepatitis: 

Results of a Markov Model 

Matthew Klebanoff 1,2 , Kathleen E. Corey 2,3 , Lee M. Kaplan 2,3 , 

Raymond T. Chung 2,3 , Chin Hur 2,3 ; 1 Institute for Technology 

Assessment, Massachusetts General Hospital, Boston, MA; 2 Gastrointestinal 

Unit, Massachusetts General Hospital, Boston, MA; 

3 Harvard Medical School, Boston, MA 

Obese individuals are at heightened risk of developing nonalcoholic 

steatohepatitis (NASH), which can give rise to cirrhosis 

and hepatocellular carcinoma. Current treatments for NASH 

are poorly tolerated or have little long-term data to support their 

use. A growing body of evidence suggests that bariatric surgery 

not only leads to weight loss but may also improve NASH. 

Using a Markov model, we determined the gain in life years 

(LYs) and quality-adjusted life years (QALYs) following laparoscopic 

Roux-en-Y gastric bypass (L-RYGB) for patients in various 

weight classes with varying degrees of NASH fibrosis (F0-F3). 

After surgery, a percentage of patients who benefit (69.7% in 

base-case, from Mathurin et al., 2009) enter a ‘NASH remission’ 

state and stop progressing toward cirrhosis, although 

they may later relapse (Fig. 1). The primary analysis does not 

model surgery in F4 patients due to a lack of data in this group. 

The model incorporated life year and quality-of-life decrements 

associated with L-RYGB and complications. Extensive sensitivity 

analyses examined the impact of model input uncertainty on 

results. For all classes of obesity (mild, moderate and severe), 

surgery led to a gain in LYs and QALYs. When we excluded 

benefits of weight loss to explore the potential impact of surgery 

on non-obese NASH patients, surgery reduced life expectancy 

by 0.02 LYs for F0 patients, but increased LYs for F1-F3 

patients. Surgery also decreased quality-adjusted survival for 

F0 and F1 patients by 0.28 and 0.22 QALYs, respectively, 

but increased QALYs for F2 and F3 patients. The number of 

F3 patients needed to treat (NNT) to prevent one liver-related 

death was seven; for F2 patients, the NNT was 17. Conclusions: 

Bariatric surgery is effective as a treatment for patients 

with NASH in all obesity classes. When we focused only on the 

benefit due to NASH remission by eliminating any weight loss 

benefit from the model, bariatric surgery improved life expectancy 

for F1-F3 patients, but increased quality-adjusted survival 

only for patients with more advanced fibrosis (F2 and F3), with 

relatively favorable NNTs. 

Disclosures: 

Kathleen E. Corey - Advisory Committees or Review Panels: Gilead; Speaking 

and Teaching: Synageva 

Lee M. Kaplan - Consulting: Johnson and Johnson, GI Dynamics, Novo Nordisk; 

Grant/Research Support: Ethicon 


Merck, BMS 

The following authors have nothing to disclose: Matthew Klebanoff, Chin Hur 

155 

A Trial-based Model of Liver Transplant and Liver-related 

Death in Patients with Primary Biliary Cirrhosis 

(PBC) 

Marco Carbone 1 , Richard Pencek 2 , Tracy J. Mayne 2 , Tonya Marmon 

2 , George F. Mells 3 , David Shapiro 2 ; 1 Division of Gastroenterology 

and Hepatology, Department of Medicine, Addenbrooke’s 

Hospital, Cambridge, United Kingdom; 2 Intercept Pharmaceuticals, 

Inc., San Diego, CA; 3 Division of Gastroenterology and 

Hepatology, Department of Medicine, University of Cambridge, 

Cambridge, United Kingdom 

Background: The UK-PBC research group derived and validated 

a predictive model for liver transplant or liver-related 

death in PBC patients based on Cox proportional hazards 

regression analysis of the UK-PBC cohort (n=4022). Predictors 

included ALP, bilirubin, ALT, albumin and platelet count. Obeticholic 

acid (OCA) is a selective FXR agonist in development for 

the treatment of PBC. In a recent Phase 3 trial (POISE), OCA 

treatment was associated with significant improvements in ALP, 

bilirubin and transaminases compared to placebo +UDCA. 

Objective: To apply the UK-PBC risk algorithm to the POISE 

trial data to predict effect of OCA on liver transplant and liver-related 

death. Methods: In POISE, 216 PBC patients with 

inadequate response or intolerance to UDCA were randomly 

assigned to: OCA 10 mg, OCA titration (starting at 5 mg and 

adjusted to 10 mg at 6 months based on clinical response) or 

placebo; pre-trial UDCA continued. Treatment effect on liver 

biochemistry was assessed at 12 months. The UK-PBC algorithm 

assessed risk for liver-transplant or liver-related death at 

5, 10 and 15 years based on 12-month change from baseline. 

Results: The demographics of the POISE cohort were: mean age 

56 years, 91% female, 94% white, 93% on UDCA. At baseline, 

the UK-PBC algorithm indicated that placebo patients were 

at slightly higher risk for events compared to both OCA arms. 

At 12 months, predicted risk had significantly improved with 

OCA, primarily due to improved ALP and bilirubin. Risk worsened 

in the placebo arm, primarily due to increased bilirubin. 

Predicted risk at 5, 10 and 15 years is shown below. Based on 

absolute risk reduction at 15 years, the number needed to treat 

with OCA to avoid liver transplant or liver-related death was 

13 (OCA 10 mg) and 11 (OCA titration). Conclusions: The 

UKPBC risk model is a validated prognostic indicator of liver 

transplant-free survival. When applied to patients in POISE,


the UK-PBC algorithm predicts a significant decrease in risk for 

liver transplant and liver-related death in patients treated with 

OCA+UDCA relative to placebo+UDCA. OCA has the potential 

to be an important new treatment option for PBC patients 

unable to achieve adequate response with UDCA. 

Disclosures: 

Richard Pencek - Employment: Intercept Pharmaceuticals; Stock Shareholder: 

Intercept Pharmaceuticals 






The following authors have nothing to disclose: Marco Carbone, George F. Mells 

156 

Costs per Diagnosed Liver Disease Stage Among Individuals 

with Chronic Hepatitis C Virus in the Veterans 

Health Administration 

Jennifer R. Kramer 1 , Peter Richardson 1 , Danielle Liffmann 2 , Hashem 

B. El-Serag 1 , David B. Rein 2 ; 1 Department of Medicine, Michael E. 

DeBakey VA Medical Center and Baylor College of Medicine, 

Houston, TX; 2 NORC at the University of Chicago, Chicago, IL 

Background: The prevalence of hepatitis C virus (HCV) infection 

in the Veteran Health Administration (VHA) is high, but 

previous research has not evaluated the costs of medical care 

for diagnosed patients. Methods: We used the VHA HCV 

Clinical Case Registry and Health Economics Resource Center 

average cost data of inpatient, outpatient, and long-term care 

to estimate the incremental annual cost of diagnosed HCV. 

We compared the total healthcare costs of HCV antibody and 

RNA + patients (cases) to HCV antibody + but HCV RNA - 

patients (controls) identified from FY 1999 to 2009. Cases 

were matched 1:1 to all available controls using a propensity 

score that included demographic and clinical characteristics. 

Patients were required to have an available AST/platelet ratio 

(APRI) value in the index year (year of the earliest RNA test) 

and one subsequent year. In each year, we classified patients 

using ICD-9 codes and lab data (highest APRI in year) into 8 

clinical stages: APRI 2, decompensated 

cirrhosis (DCC), hepatocellular carcinoma (HCC), 

transplant/post-transplant care, and death while infected with 

HCV. Patients with DCC or HCC prior to HCV index date 

were excluded. We estimated the incremental annual costs 

for the 6 years after index date of each stage of HCV using 

a hierarchical longitudinal two-part health expenditure model 

(logistic, generalized linear model assuming a gamma distribution 

and log link) and adjusted for clustering by incorporating 

patient level random effects in both the logistic and gamma 

models allowing these effects to be correlated. Results: There 

were 157,303 patients (140,169 cases/17,134 controls) in 

our source population; 52.8 years old (sd=8.0), 50.6% white, 

25.9% Black, 96.7% male. The analysis set included 17,134 

cases that were propensity matched to the 17,134 controls. 

In 2009, 52.1% had APRI 2, 5.6% DCC, 0.7% HCC, 0.1% transplant, 

and 3.9% died. Using 2015 dollars, on average, HCV RNA 

- patients with an APRI


with the new balloon score. Most showed a stepwise increase 

across the range of the score. Conclusion: The new balloon 

score shows excellent correlation with clinical disease features. 

Replacement of the old balloon score with the new balloon 

score in the NAS would give more weight to ballooning as well 

as increasing the dynamic range of the NAS. 

NASH CRN New Balloon Score 

Disclosures: 

Elizabeth M. Brunt - Consulting: Synageva; Independent Contractor: Rottapharm 

Brent A. Neuschwander-Tetri - Advisory Committees or Review Panels: Nimbus 

Therapeutics, Bristol Myers Squibb, Janssen, Mitsubishi Tanabe, Conatus, 

Scholar Rock 






The following authors have nothing to disclose: David E. Kleiner, Patricia H. Belt, 

Cynthia A. Behling, Ryan M. Gill, Cynthia D. Guy, Mark L. Van Natta 

158 

Overweight in late adolescence is associated with 

decompensated liver disease later in life after 39 years 

of follow-up 

Hannes Hagström 1 , Per Stål 1 , Rolf W. Hultcrantz 1 , Tomas Hemmingsson 

2,3 , Anna Andreasson 4,5 ; 1 Center for Digestive Diseases, 

Unit of Hepatology, Karolinska University Hospital. Department 

of Medicine, Huddinge, Karolinska Institutet, Stockholm, Sweden, 

Stockholm, Sweden; 2 Institute of Environmental Medicine, Karolinska 

Institutet, Stockholm, Sweden; 3 Centre for Social Research 

on Alcohol and Drugs, Stockholm University, Stockholm, Sweden; 

4 Stress Research Institute, Stockholm University, Stockholm, Sweden; 

5 Division of Family Medicine, Karolinska Institutet, Stockholm, 

Sweden 

Background: The prevalence of both obesity and liver diseases 

is increasing globally. Obesity is associated with a worse prognosis 

in different liver diseases. If overweight and obesity in 

late adolescence is an independent predictor of liver disease 

later in life is not studied. We investigated if overweight per se 

predicts development of liver disease and decompensated liver 

disease later in life with a up to 39 year follow-up. Materials 

and methods: Data from 49,321 men (18-21 years) conscribed 

to military service in Sweden between 1969 and 1970 was 

used. Conscription during this time was mandatory and >97% 

of the male population was available for this study. Weight and 

height measured at conscription were used to calculate body 

mass index (BMI). Data was collected from the national patient 

register to identify any diagnosis of liver disease at a Swedish 

hospital from time of conscription until the end of 2009. 

A multivariate logistic regression model was used to estimate 

odds ratios (OR) of decompensated liver disease (hepatorenal 

syndrome [HRS], hepatocellular carcinoma [HCC], ascites, 

varices or hepatic encephalopathy [HE]) for persons with BMI 

over as compared to under 25. The model was adjusted for 

alcohol use, smoking, use of narcotics, social status and blood 

pressure at the time of conscription. Results: Mean BMI at conscription 

was 20.97 kg/m 2 , where 6.6% had a BMI > 25 and 

0.8% had a BMI > 30. During a follow-up period of in mean 

37.9 (+/-4.8 years, range 0-39) years of follow-up, 525 persons 

were diagnosed with liver disease. Of these, 206 persons 

developed decompensated liver disease (18 cases of HRS, 31 

HCC, 79 ascites, 124 varices and 7 HE as first manifestation 

of decompensation, where 43 persons had multiple diagnoses 

at presentation). Mean time to decompensation was 29.8 

years (95% Confidence Interval [CI] 28.8-30.8 years). BMI > 

25 was significantly associated with development of decompensated 

liver disease both in the univariate (OR 2.09, 95%CI 

1.38-3.16, p


than children with Zone 3 steatosis (73% vs 58%, p


olites predictive of fibrosis stage. RESULTS: Age, gender, BMI, 

current/past alcohol use/smoking, total HDL/LDL cholesterol 

or triglycerides did not differ with fibrosis stage. In univariate 

testing, plasma concentrations of alpha-glutamyltyrosine N-acetylcitrulline 

palmitoyl-palmitoyl-glycerophosphocholine, glycocholate 

glutarate, fucose and fumarate associated with fibrosis 

stage (p=0.0007). After controlling for age, gender and DM, 

N-acetylcitrulline, alpha glutamytyrosine, palmitoyl-palmitoyl-glycerophosphocholine, 

lignoceric acid, glycocholate, and 

1-arachidonoyl associated with fibrosis (p=0.0009). Multivariate 

analysis using logistic regression yielded an Area Under 

the Receiver Operating Characteristic (AUROC) of 0.8804 

for I-urobilinogen, malate, glutarate (pentanedioate), taurochenodeoxycholate, 

3-hydroxyoctanoate, fucose, isoleucine, 

palmitoyl-palmitoyl-glycerophosphocholine, N-acetylcitrulline. 

Sparse ordinal regression was used to predict fibrosis stage, 

resulting in a correlation coefficient of 0.3757 (p=5x10 -8 ) 

after leave-one-out cross-validation. The model identified a 

69 metabolite signature predictive of fibrosis stage. CONCLU- 

SIONS: Analysis of serum from NAFLD patients revealed alterations 

in bile acids, steroids hormones, branched-chain amino 

acid catabolism, TCA cycle metabolism, and mitochondrial 

function in stage 3-4 fibrosis. The role of these metabolites as 

potential biomarkers for hepatic fibrosis requires validation. 

Pathways involved in bile acid and/or steroid hormone metabolism, 

branched-chain amino acids, and/or mitochondrial function 

may offer novel targets for anti-fibrotic therapies in NAFLD. 

Disclosures: 

Lauren N. Bell - Employment: Metabolon, Inc. 

Regis Perichon - Employment: Metabolon 




The following authors have nothing to disclose: Ricardo Henao, Nigar Hasanaliyeva, 

Jacob Wulff, Cynthia A. Moylan, James T. Lu, Cynthia D. Guy, Anna Mae 

Diehl 

162 

Beneficial effects of the dual PPAR α-δ agonist, GFT505, 

on hepatic and cardiometabolic markers in adult NASH 

patients. 

Stephen A. Harrison 3 , Arun J. Sanyal 2 , Sven M. Francque 4 , Pierre 




Alice Roudot 13 , Sophie Megnien 13 , Bart Staels 14 , Vlad Ratziu 1 ; 

1 Hepatology, Hopital Pitie Salpetriere, Paris, France; 2 Internal 

Medicine/Division of Gastroenterology, Hepatology and Nutrition, 

Virginia Commonwealth University, Richmond, VA; 3 Gastroenterology, 

Brooke Army Medical Center, Fort Worth, TX; 4 Antwerp 

Univesrity Hospital, Gastroenterology Hepatology, Antwerp, Belgium; 

5 Pathology, Hopital Beaujon, Clichy, France; 6 Hepatology 

Department, Hospital Saint-Antoine, Paris, France; 7 Valme University 

Hospital, Digestive Diseases, Sevilla, Spain; 8 Hepatology 

Department, Centre Hospitalier Universitaire d’ Angers, Angers, 

France; 9 Medicine, Duke University, Durham, NC; 10 Hepatology 

Department, University of Virginia, Charlotesville, VA; 11 Hepatology 

Department, RUNMC, Nijmegen, Netherlands; 12 Institute 

of Cellular Medicine, Newcastle University, Newcastle, United 

Kingdom; 13 Genfit, Loos, France; 14 INSERM U1011, European 

Genomic Institute for Diabetes (EGID), Université Lille 2, Lille, 

France 

Introduction: NASH patients (pts) have a high prevalence of cardiometabolic 

risk factors, hence cardiovascular disease is the 

leading cause of death in this population. Having demonstrated 

the efficacy of GFT505 on the histological reversal of NASH, 

here we evaluated the biochemical and cardiometabolic effects 

of the drug in in this large, international, phase 2 randomized, 

controlled trial of adult pts with biopsy-proven NASH. Methods: 

Pts from the ITT population of the GOLDEN505 trial randomized 

to the GFT505 120 mg (GFT120) and placebo (PBO) 

arms were compared for treatment effects on plasma lipid, 

glycemic, insulin resistance, inflammatory and liver markers. 

Results are expressed as effect size vs. placebo (LS mean±SE). 

Analyses were conducted for different stages of disease severity 

based on the NAFLD Activity Score (NAS) at baseline (NAS 3: 

mild, NAS 4-5: moderate, NAS>5: severe), and fibrosis stage 

(F0-F1 vs. F2-F3 according to Kleiner et al). Results: Compared 

to PBO, GFT120 significantly improved GGT (-29.31±6.36 

U/L, p





The following authors have nothing to disclose: Sven M. Francque, Pierre 

Bedossa, Joost Drenth 

163 

Osteopontin is involved in chronic HBV infection and 

enhances HBV replication and HBsAg secretion. 

Sandra Phillips, Jason D. Coombes, Sameer Mistry, Roger Williams, 

Wing-Kin Syn, Shilpa Chokshi; Institute of Hepatology, 

Foundation for Liver Research, London, United Kingdom 

Background/Aims: Hepatitis B virus (HBV) requires host cellular 

machinery such as cyclophilins to support its ongoing propagation 

(Phillips et al, Gastroenterology 2014). These host proteins 

represent ideal candidates for therapeutic interventions 

as they are generally expected to have a lower frequency of 

drug-resistance and antiviral efficacy across genotypes. We 

have also recently described a role for the host protein Osteopontin 

(OPN), a pro-fibrogenic downstream effector of the 

Hedgehog pathway, in enhancing HCV replication (Choi et al, 

Clinical Sciences 2014). Whilst increased levels of blood OPN 

have similarly been reported in chronic Hepatitis B (CHB), its 

role in viral replication remains unknown. This study aimed 

to evaluate the role of OPN in HBV replication, HBsAg secretion 

and HBV-driven liver injury. Material and Methods: Stably 

(HepG2215), transiently (HUH-7) transfected and infected 

(HepaRG) cell lines, producing full HBV virions and HBsAg 

particles were cultured over 72h in the presence/absence of 

several concentrations of recombinant OPN (recOPN). In addition, 

secreted OPN was neutralized using OPN-specific aptamers. 

Cells and supernatants were harvested at baseline, 24, 48 

and 72 hours. Intracellular OPN mRNA and HBV-DNA levels 

were quantitated by Real-Time qPCR. HBsAg levels were measured 

by ELISA. OPN levels were also measured by ELISA in 

cell culture supernatants and in sera of controls and HBeAg(+) 

CHB patients who were either treatment naive or treated with 

potent antiviral agents. In addition, expression of OPN was 

assessed in explanted livers from healthy and HBV-cirrhotic 

patients. Results: Serum levels and intrahepatic expression of 

OPN were significantly increased in CHB patients compared 

to controls (up to 7 fold; p


165 

The HBx-DLEU2 lncRNA complex regulates transcription 

from cellular genes and the HBV cccDNA 

Francesca Guerrieri 1,2 , Letizia Chiodo 1 , Debora Salerno 1,2 , Safaa 

Jeddari 2 , Giancarlo Ruocco 1 , Massimo Levrero 2,1 ; 1 CLNS@SAPI- 

ENZA, Istituto Italiano di Tecnologia (IIT), Rome, Italy; 2 Dept of 

Internal Medicine, La Sapienza University, Rome, Italy 

Background: HBx regulatory protein is required for HBV 

cccDNA transcription/viral replication and contributes to HBV 

oncogenicity. HBx affects the epigenetic control of HBV viral 

chromatin, by preventing HDACs and PMRT1 recruitment onto 

the cccDNA, as well as of cellular chromatin, by modulating 

the recruitment of chromatin modifying enzymes. We previously 

showed that HBx binds to the regulatory regions of several 

ncRNAs (75 miRNAs and 34 lncRNAs). DLEU2 lncRNA 

overlaps with the autophagy TRIM13 gene in the opposite 

orientation. Upregulation of specific DLEU2 splicing variants 

correlates with the development of solid and onco-hematolocic 

malignancies. Objectives: Aim of this study is to clarify the role 

of HBx in the regulation of the DLEU2/TRIM13 transcriptional 

unit and their functions. Methods: The anti-HBx ChIPSeq was 

performed in mock and HBV-replicating HepG2 cells using an 

Illumina NGS platform. Independent ChIPs were analyzed by 

TaqMan real-time PCR using lncRNA and gene specific primers. 

The nCounter Nanostring technology and real-time RT-PCR 

were used to evaluate lncRNAs and target gene expression, 

respectively. Specific LNA longRNA GapmeRs (Exiqon) were 

used for highly efficient inhibition of DLEU2 lncRNA function. 

Results: ChIPSeq analysis of HBx global chromatin recruitment 

revealed a specific binding to 34 lncRNAs regulatory 

regions. Using a custom Nanostring panel we found that all 

putative lncRNAs targets were modulated in HBV-replicating 

HepG2 cells. Focusing on DLEU2 lncRNA, we demonstrated 

that HBx is able to deregulate both its expression and the overlapping 

gene TRIM13. HBx binding to the DLEU2 promoter 

affects its epigenetic control, changes DLEU2 splicing profile 

and up-regulates the antisense gene TRIM13. Selective degradation 

of DLEU2 RNA resulted in a reduced H4 acetylation 

on the TRIM13 promoter and a ~50% reduction of TRIM13 

expression in HBV replicating HepG2 cells. In silico analysis of 

a model HBx protein tertiary structure and DLEU2 tertiary structure 

suggested a direct interaction. Using a RIP (RNA Immune 

Precipitation) approach we confirmed HBx-DLEU2 interaction 

in vivo. Finally, we found that DLEU2 inactivation inhibits the 

expression of the HBx targets SREBP2 and miR138 as well as 

HBV cccDNA transcription, with a sharp decrease of pgRNA 

levels, thus suggesting a functional relevance of the DLEU2- 

HBx interaction of HBV replication. Conclusion: HBx binds to 

the DLEU2 promoter region to modify its epigenetic regulation 

change the DLEU2 splicing profile. HBx forms a functional complex 

with DLEU2 and regulates HBV replication. 

Disclosures: 

Massimo Levrero - Advisory Committees or Review Panels: BMS, Jansen, Gilead, 

Tekmira, Galapagos, Medimmune; Speaking and Teaching: MSD, Roche 

The following authors have nothing to disclose: Francesca Guerrieri, Letizia Chiodo, 

Debora Salerno, Safaa Jeddari, Giancarlo Ruocco 

166 

Viral Expression and Molecular Profiling of Liver Tissue 

and Microdissected Hepatocytes in Hepatitis D Virus 

(HDV) - Associated Hepatocellular Carcinoma (HCC) 

Patrizia Farci 1 , Ashley B. Tice 1 , Ronald E. Engle 1 , Fausto Zamboni 

2 , Marta Melis 1 , Zhifeng Long 3 , Jaime Rodriguez-Canales 4 , 

Jeffrey Hanson 4 , Michael R. Emmert-Buck 4 , David E. Kleiner 4 , 

Giacomo Diaz 5 ; 1 Laboratory of Infectious Diseases, National Inst 

of Health, Bethesda, MD; 2 Liver Transplantation Center, Brotzu 

Hospital, Cagliari, Italy; 3 Personal Diagnostix, Gaithersburg, MD; 

4 Laboratory of Pathology, National Cancer Institute, Bethesda, 

MD; 5 Department of Biomedical Sciences, University of Cagliari, 

Cagliari, Italy 

Although HCC develops in a high proportion of patients 

with chronic hepatitis D, there are no data on the molecular 

mechanisms of HDV-induced hepatocarcinogenesis. Because 

of the vital dependence of HDV on HBV, the role of HDV in 

promoting HCC is unknown. We used genomic techniques 

to dissect the role of HDV and HBV. Microarray (Affymetrix 

Human U133 Plus2) was performed on 33 specimens of 

whole liver tissue (WLT, tumor vs. non-tumor) and selected 

laser capture-microdissected hepatocytes (LCM, malignant vs. 

non-malignant hepatocytes) obtained at liver transplant (LT) 

from 5 patients with HDV-HCC; 29 WLT specimens from 7 

patients with non-HCC HDV cirrhosis served as controls. A 

parallel analysis was performed on WLT and LCM from 11 

patients with HBV-HCC without HDV. Microarray data were 

analyzed using BRB-Array Tools, with multivariate permutation 

F-tests (FDR


167 

Characterisation of the immune profile in Chronic Hepatitis 

B, with CyTOF, to identify biomarkers of immune 

control following NUC therapy discontinuation 

Upkar S. Gill 1 , Laura Rivino 2 , Nina Le Bert 3 , Kamini Kunasegaran 

2 , Damien Tan 3 , Sarene Koh 3 , Machteld Van Den Berg 2 , Yang 

Cheng 4 , Navjyot K. Hansi 1 , Graham R. Foster 1 , Evan W. Newell 

4 , Antonio Bertoletti 2,3 , Patrick T. Kennedy 1 ; 1 Hepatology Unit, 

Centre for Immunobiology, Blizard Institute, Barts and The London, 

School of Medicine & Dentistry, QMUL, London, United Kingdom; 

2 Program Emerging Viral Diseases, Duke-NUS Graduate Medical 

School, Singapore, Singapore; 3 Infection & Immunity Program, 

Singapore Institute for Clinical Sciences, Agency for Science, Technology 

& Research (A*STAR), Singapore, Singapore; 4 Singapore 

Immunology Network, Singapore Agency for Science, Technology 

& Research (A*STAR), Singapore, Singapore 

INTRODUCTION: The absence/functional exhaustion of 

HBV-specific T cells is the hallmark of chronic HBV infection 

(CHB); conversely a robust functional response of these cells 

is associated with viral control. Current therapies are limited 

in their ability to restore the functional HBV-specific repertoire. 

Consequently, Nucleot(s)ide analogue (NUC) therapy remains 

indefinite in the majority. Clinical parameters alone cannot 

distinguish in which patients NUC therapy can be safely discontinued, 

with durable immune control from those who will 

relapse and develop hepatic flares (HF). We studied a cohort 

of virally suppressed patients on potent NUC therapy prior to 

and after treatment discontinuation to characterise the immune 

profile associated with viral control. PATIENTS & METHODS: 

PBMC were analysed at 4-weekly intervals prior to and after 

NUC discontinuation. The frequency of HBV-specific T cells 

was assessed by IFNγ ELISPOT after 10 day expansion in the 

presence of HBV peptides spanning the entire viral proteome. 

Phenotypic and functional characteristics of T & NK cells were 

studied with an in-depth longitudinal analysis of the expression 

of >35 markers involved in cell activation, differentiation and 

exhaustion, by cytometry time of flight (CyTOF), a novel mass 

cytometry technology. Serum cytokine and chemokine levels 

(IL-1β, IL-6, TNF-α, IL-10, CXCL-8, CXCL-10) were also measured 

using Luminex. RESULTS: Prior to NUC discontinuation, 

patients could be divided into 2 groups based on the presence/absence 

of HBV-specific T cells. Upon NUC discontinuation, 

those patients with detectable frequencies of circulating 

HBV-specific T cells were able to control HBV replication and 

did not demonstrate HF’s. In these patients HBV-specific T cells 

preferentially target HBV polymerase followed by core proteins. 

In patients without viral control who developed HF, we 

noted a significant increase in serum CXCL10 and IL10 levels, 

correlating with ALT. Patients without HF’s were characterised 

by increased frequencies of a CD8+ T cell subset expressing 

CD56, CD107, CCR5, CD127, CD28, 2B4, KLRG1, IFNγ, 

TNFα, MIP1β & IL2. NK cell profiles did not differ depending 

on viral rebound/HF upon NUC discontinuation, but all CHB 

patients expressed significantly higher levels of the exhaustion 

marker, KLRG1 on NK cells, compared with healthy controls. 

CONCLUSIONS: Our data suggest that HBV-polymerase 

specific T cells represent a potential biomarker that can be 

used to predict those patients who will control HBV after NUC 

discontinuation. These results further support the concept that 

HBV-specific T cells are associated with viral control rather than 

contributing to immunopathology. 

Disclosures: 

Graham R. Foster - Advisory Committees or Review Panels: GlaxoSmithKline, 

Novartis, Boehringer Ingelheim, Tibotec, Chughai, Gilead, Janssen, Idenix, 

GlaxoSmithKline, Novartis, Roche, Tibotec, Chughai, Gilead, Merck, Janssen, 

Idenix, BMS; Board Membership: Boehringer Ingelheim; Grant/Research Support: 

Chughai, Roche, Chughai; Speaking and Teaching: Roche, Gilead, Tibotec, 

Merck, BMS, Boehringer Ingelheim, Gilead, Janssen 

Patrick T. Kennedy - Grant/Research Support: Roche, Gilead; Speaking and 

Teaching: BMS, Roche, Gilead 

The following authors have nothing to disclose: Upkar S. Gill, Laura Rivino, Nina 

Le Bert, Kamini Kunasegaran, Damien Tan, Sarene Koh, Machteld Van Den Berg, 

Yang Cheng, Navjyot K. Hansi, Evan W. Newell, Antonio Bertoletti 

168 

Hepatitis B virus X protein stimulates HBV replication by 

regulating transcription factors associated with DNA or 

histone methylation 

Naoki Oishi 1,2 , Xuyang Wang 2 , Kazunori Kawaguchi 1,2 , Masao 

Honda 1,2 , Seishi Murakami 2 , Shuichi Kaneko 1,2 ; 1 Department of 

Gastroenterology, Kanazawa University Hospital, Kanazawa, 

Japan; 2 Department of Disease Control and Homeostasis, 

Kanazawa University Hospital, Kanazawa, Japan 

Background: Hepatitis B virus (HBV), a small enveloped DNA 

virus, chronically infects more than 350 million people worldwide 

and causes liver diseases, from hepatitis to cirrhosis and 

liver cancer. HBx is a multifunctional protein encoded by the 

HBV genome that stimulates HBV replication. Previously, we 

found that HBx has an important role in stimulating HBV transcription 

and replication and that the transcriptional transactivation 

function of HBx may be critical for its augmentation 

effect on HBV replication. However, the molecular mechanism 

of HBx in transcriptional coactivation remains unclear. In this 

study, we provide a new insight into the coactivator mechanism 

and the possibility of a new treatment target for HBV replication 

and HBV-related hepatocarcinogenesis. Methods: We 

used a retroviral vector to introduce wild-type HBx (HBxwt) or 

empty vector (EV) into HepG2 cells. Gene expression profiling 

was performed using Affymetrix GeneChip Human U133A2.0 

ver.2.0 arrays according to the manufacturer’s protocol. Unsupervised 

hierarchical clustering analysis and class comparison 

analysis were performed with BRB-Array Tools software version 

4.2.2. Transcription factor analysis was performed using 

Ingenuity Pathway Analysis (IPA; Ingenuity Systems) to identify 

potential upstream transcription factors (TFs). We used array 

data from 244 chronic hepatitis B patients for analyzing clinical 

features. Results: Unsupervised hierarchical clustering analysis 

of HepG2-HBxwt or HepG2-EV cells revealed that the gene 

expression profiles of these cell lines were significantly different. 

Class comparison analysis between both cell lines identified 

803 HBx-related genes. Six HBx-related activated TFs that 

affected HBV replication were identified by IPA, small interfering 

RNA, and inhibitor analyses. Three of these six TFs control 

HBV replication by modifying DNA or histone methylation. By 

analysis of 244 hepatitis B patients, these three TFs were found 

to be expressed at a significantly highly level in HBeAg(+) 

HBeAb(-) cases than in HBeAg(-)HBeAb(+) cases. Moreover, 

these TFs up-regulated the expression of stemness markers 

(EpCAM, AFP, and SOX9) and epithelial-mesenchymal transition 

markers (ZEB1, ZEB2, and VIM) and were associated with 

a poor prognosis of HCC. Conclusions: HBx upregulated three 

TFs associated with DNA or histone methylation. Moreover, 

HBx stimulates HBV replication and hepatocarcinogenesis by 

modifying DNA or histone methylation. Our study suggests that 

TFs activated by HBx will be an important therapeutic target 

against both virus replication and hepatocarcinogenesis aimed 

at the prevention of HCC.


Disclosures: 






The following authors have nothing to disclose: Naoki Oishi, Xuyang Wang, 

Kazunori Kawaguchi, Masao Honda, Seishi Murakami 

169 

DNA Methylation Markers for Detection of Extrahepatic 

Cholangiocarcinoma: Discovery, Tissue validation, and 

Pilot Testing in Biliary Brush Samples 

Hassan M. Ghoz 1 , Mohammed M. Aboelsoud 5 , Tracy C. Yab 1 , 

Calise K. Berger 1 , William R. Taylor 1 , Xiaoming Cao 1 , Patrick 

H. Foote 1 , Nasra H. Giama 1 , Catherine D. Moser 1 , Douglas W. 

Mahoney 4 , Emily G. Barr Fritcher 2 , Benjamin R. Kipp 3 , Thomas C. 

Smyrk 3 , Lewis R. Roberts 1 , David Ahlquist 1 , John B. Kisiel 1 ; 1 Division 

of Gastroenterology and Hepatology, Mayo Clinic, Rochester, 

MN; 2 Molecular Anatomic Pathology, Mayo Clinic, Rochester, 

MN; 3 Anatomic Pathology, Mayo Clinic, Rochester, MN; 4 Biomedical 

Statistics and Infomatics, Mayo Clinic, Rochester, MN; 

5 Memorial Hospital, Pawtucket, RI 

Background & Aims: Cholangiocarcinoma (CCA) has poor 

prognosis due to late stage presentation. Molecular markers 

may offer improved accuracy for early detection. We have 

identified discriminant DNA methylation markers for intrahepatic 

CCA by next-generation sequencing. We now explore 

this discovery approach further with extrahepatic CCA (eCCA) 

and pilot best markers on brush cytology specimens. Methods: 

Reduced-representation bisulfite sequencing (RRBS) was performed 

on DNA extracted from 18 frozen eCCA tissue samples 

and matched, adjacent benign biliary epithelia or liver parenchyma. 

Differentially methylated regions (DMRs) with at least 

3 CpGs were ranked by area under the receiver operating 

characteristics curve (AUC) & by tumor:normal ratio and then 

technically validated by methylation specific PCR (MSP) on 

DNA from same samples. Best DMRs were selected for biological 

validation on DNA from independent tissues comprising 

15 eCCA cases and 60 controls (6 adjacent bile duct, 18 

adjacent liver, 18 white blood cell samples, 18 normal colon 

epithelia) using MSP. Biologically valid DMRs were then blindly 

assayed on DNA extracted from independent archival biliary 

brushing specimens including 14 perihilar (pCCA) & 4 distal 

(dCCA) cases and 18 matched cytology-negative controls 

(CTRL), 4 of which had primary sclerosing cholangitis (CTRL- 

PSC). Results: From 5.5 million CpGs, 3674 significant DMRs 

were mapped; 46 were selected for technical validation from 

which 18 DMRs had an AUC of 0.75–1.0. In biological validation, 

8 of these showed an AUC >0.75 in eCCA tissues. In 

brushings, methylated EMX1, HOXA1, VSTM2B.764, KC01, 

BMP3, SALL1, PTGDR, and RYR2, showed sensitivities of 100% 

89%, 83%, 78%, 72%, 72%, 72%, and 72%, respectively, 

at 90% specificity. Accuracy of EMX1 on brushings is shown 

(figure). Conclusion: Whole-methylome discovery by next-generation 

DNA sequencing yielded novel, highly-discriminant 

methylation markers for eCCA. Results were validated in independent 

tissues as well as cytology brushings. Further clinical 

evaluation is clearly warranted. 

Disclosures: 

Tracy C. Yab - Patent Held/Filed: Exact Sciences; Stock Shareholder: Exact 

Sciences 

William R. Taylor - Patent Held/Filed: Exact Sciences; Stock Shareholder: Exact 

Sciences 

Douglas W. Mahoney - Patent Held/Filed: Exact Sciences 

Benjamin R. Kipp - Grant/Research Support: Abbott Molecular Inc. 




David Ahlquist - Advisory Committees or Review Panels: exact sciences; Consulting: 

exact sciences; Grant/Research Support: exact sciences; Stock Shareholder: 

exact sciences 

John B. Kisiel - Grant/Research Support: Exact Sciences Corporation 

The following authors have nothing to disclose: Hassan M. Ghoz, Mohammed M. 

Aboelsoud, Calise K. Berger, Xiaoming Cao, Patrick H. Foote, Nasra H. Giama, 

Catherine D. Moser, Emily G. Barr Fritcher, Thomas C. Smyrk 

170 

Circulating Osteopontin and Prediction of Hepatocellular 

Carcinoma Development in a Large European Population 

Talita Duarte-Salles 2 , Sandeep Misra 1 , Magdalena Stepien 2 , 

Mazda Jenab 2 , Pierre Hainaut 2 , Laura Beretta 1 ; 1 Molecular and 

Cellular Oncology, MD Anderson Cancer Center, Houston, TX; 

2 International Agency for Research on Cancer (IARC-WHO), Lyon, 

France 

Background: Prevention and early detection strategies are 

urgently needed to reduce the burden and mortality of hepatocellular 

carcinoma (HCC), the second leading cause of cancer 

death worldwide. We previously identified osteopontin 

(OPN) as a promising marker for the early detection of HCC. 

In this study, we investigated the association between pre-diagnostic 

circulating OPN levels and HCC incidence and OPN 

performance for early detection of HCC in a large population-based 

cohort. Methods: A nested-case control study was 

conducted within the European Prospective Investigation into 

Cancer and Nutrition (EPIC) cohort study, a large longitudinal 

cohort of >520,000 participants from 10 Western European 

countries. During a mean follow-up of 4.8 years, 100 HCC 

cases were identified. Each case was matched to two controls 

by incidence density sampling. OPN levels were measured in 

baseline plasma samples. Hepatitis infection status and other 

relevant biomarkers such as alpha-fetoprotein (AFP) were also 

measured. Conditional logistic regression models were used to 

calculate multivariable odds ratio (OR) and 95% confidence 

intervals (95%CI) for tertiles and continuous OPN levels in relation 

to HCC. Receiver operating characteristics curves (ROC) 

were constructed to determine the discriminatory accuracy of 

OPN alone or in combination with other liver function biomarkers 

in the prediction of HCC. Behavior of OPN in relation to 

time of HCC diagnosis was also evaluated. Results: Circulating 

OPN level was significantly associated with increased HCC 

risk (per 10% increment in OPN level, OR multivariable 

=1.30;


95%CI: 1.14-1.48), also among hepatitis-free participants 

(OR multivariable 

=1.19; 95%CI: 1.05-1.34). Adding liver function 

tests to OPN improved the discrimination of those subjects who 

developed HCC (AUC=0.86) while AFP didn’t improve the 

model. In contrast, within two years of HCC diagnosis, the 

combination of OPN and AFP was best able to predict HCC 

risk (AUC=0.88) while further adjustment for liver function tests 

or chronic hepatitis B/C infection did not improve the model. 

Conclusions: In this study, pre-diagnostic serum OPN concentration 

was associated with higher risk of first incident HCC 

and the association was stronger among cases diagnosed 

during the first two years of follow-up. OPN in combination 

with liver function tests improved the detection of HCC in this 

low-risk population. However, OPN in combination with AFP 

was the best predictive model for detection of HCC within 

two years of follow-up and did not change substantially after 

excluding hepatitis positive participants, or after adjustment for 

biomarkers of liver function. 

Disclosures: 

The following authors have nothing to disclose: Talita Duarte-Salles, Sandeep 

Misra, Magdalena Stepien, Mazda Jenab, Pierre Hainaut, Laura Beretta 

171 

Risk factors of complications in liver adenomatosis. 

Barbara Willandt 1 , Ragna Vanslembrouck 2 , Raymond Aerts 3 , 

Baki Topal 3 , Wim Laleman 1 , David Cassiman 1 , Schalk Van Der 

Merwe 1 , Werner Van Steenbergen 1 , Chris Verslype 1 , Frederik 

Nevens 1 ; 1 Hepatology, UZ Leuven, Leuven, Belgium; 2 Radiology, 

UZ Leuven, Leuven, Belgium; 3 Adominal Surgery, UZ Leuven, Leuven, 

Belgium 

Introduction & aims: Hepatic adenomatosis is a disorder characterized 

by multiple adenomas in an otherwise normal liver, 

originally defined as a distinct entity from a solitary hepatocellular 

adenoma. Due to the rarity of the disorder there is 

lack of consensus about treatment strategies. We explored the 

risk factors of complications and whether the genotype-phenotype 

classification used for solitary hepatic adenomas has the 

same prognostic significance in this disorder. Methods: We 

re-analyzed 35 patients with liver adenomatosis : > 10-20 

nodules (n=23pts) and > 20 (n=12pts); 6 patients with 5-10 

nodules were evaluated separately; 2 pts excluded because 

of glycogen storage disease. Biopsies, taken in the largest 

nodules, were available in 30 patients. Results: The median 

follow-up period was 6 years (range 1-24 years). All patients 

(median age 38 year, range 18-54) were female except one 

and 91% of them had a history of long term use of oral contraceptives 

(median 17 years, range 5-34). BMI was > 25 kg/m 2 

in 61% of patients and steatosis of the surrounding liver tissue 

was present in 60%. Histological subtype analysis showed: 17 

patients (63%) with inflammatory adenomas - in one patient 

associated with β-catenin activated lesions, 9 patients (33%) 

with HNF1-α mutation positive lesions of which 2 patients also 

had a β-catenin activated lesion and in 1 patient only β-catenin 

activated adenomata were identified. Inflammatory adenomas 

were clearly associated with high BMI (> 25 kg/m 2 ) and steatosis 

(13/16 (p


The following authors have nothing to disclose: Barry Schlansky, Willscott E. 

Naugler 

173 

Vitamin K dosing during sorafenib treatment for hepatocellular 

carcinoma markedly prolonged overall survival 

as well as progression free survival probably by 

augmented ischemic tumor cell damage 

Yoshimichi Haruna, Atsuo Inoue; Department of Gastroenterology 

and Hepatology, Osaka General Medical Center, Osaka, Japan 

Backgrounds and Aims. Sorafenib is the only oral anticancer 

agent for advanced hepatocellular carcinoma (HCC). However, 

its anticancer effects are not satisfying. It was reported 

that combination of vitamin K and sorafenib drastically inhibited 

HCC growth in vitro and in animal experiments. In this 

study, we examined advantage of vitamin K dosing during 

sorafenib treatment for HCC, focusing on different manners 

of changes in serum des-γ-carboxy prothrombin (DCP) levels 

during vitamin K combination and sorafenib alone groups. 

Patients and Methods. Fifty-five patients (male/female 42/13, 

age (mean ± SD) 73.9 ± 8.3, the Barcelona Clinic Liver Cancer 

(BCLC) stage B/C 21/34) were retrospectively studied. 

Twenty out of them were orally given vitamin K2 (45mg daily) 

during sorafenib treatment. There were no relevant differences 

between the vitamin K combination and sorafenib alone groups 

in characteristics at baseline. We compared the time to radiologic 

progression and overall survival (OS) in the two groups. 

The radiologic assessment was performed according to modified 

RECIST. Serum DCP levels was tested before and 8 weeks 

after the start of treatment. Results. The disease control rate was 

78.9% vs. 22.9% in sorafenib + vitamin K group and sorafenib 

alone one, respectively (P < 0.001). Median progression free 

survival (PFS) was 8.0 months vs. 2.5 months in sorafenib + 

vitamin K group and sorafenib alone one, respectively (P < 

0.001). Furthermore, vitamin K dosing markedly prolonged OS 

(median survival: 23.0 months vs. 10.0 months, P = 0.002). 

Interestingly, serum DCP levels were increased in partial 

response (PR) + stable disease (SD) cases of sorafenib alone 

group (2.42±0.92 Ç 2.78±0.95 Log mAU/mL, P = 0.051) 

despite suppressed tumor growth. In contrast, those of vitamin 

K-dosing group showed remarkable decline (2.35±0.56 

Ç 1.38±0.20 Log mAU/mL, P = 0.001). Discussion. DCP, 

a tumor marker, is also known as an indicator of ischemic 

status of HCC. The DCP elevation observed in sorafenib alone 

group seems to show ischemic status of tumor cells in deteriorated 

angiogenesis caused by sorafenib. On the other hand, 

there are many reports suggesting that the DCP is autologous 

growth factor augmenting HCC growth and a paracrine factor 

enhancing tumor angiogenesis. Thus, DCP elevation resulting 

from sorafenib treatment could incomplete the antitumor effect. 

In contrast, the suppressed DCP production under pharmacological 

vitamin K dosing could complete antitumor action of 

sorafenib. Conclusion. The nontoxic agent, vitamin K, dosing 

during sorafenib treatment for HCC markedly prolonged OS 

as well as PFS probably by augmented ischemic damage of 

tumor cells. 

Disclosures: 

The following authors have nothing to disclose: Yoshimichi Haruna, Atsuo Inoue 

174 

Outcomes following Radiofrequency Ablation of small 

HCC: Impact of etiology 

Suraj Sharma 2 , Katherine Pratte 1 , Matthew Kowgier 2 , Morris Sherman 

2 ; 1 General Internal Medicine, University of Toronto, Toronto, 

ON, Canada; 2 Toronto Liver Clinic, Toronto Western Hospital, 

Toronto, ON, Canada 

Background: Among patients who have undergone surgical 

resection, etiology of HCC may be an important predictor of 

transplant-free survival (TFS), with some studies showing worse 

survival in HCV, compared to HBV. However, there is a paucity 

of data on the impact of etiology on outcomes in patients 

undergoing RFA. This study aims to characterize a cohort of 

patients undergoing RFA for small HCCs, and determine the 

impact of etiology of HCC on TFS. Methods: Chart review was 

conducted on all patients undergoing RFA at the University 

Health Network between January 1, 2008 and December 31, 

2011. Patients were followed for outcomes until January 1, 

2014. Data collected included etiology of chronic liver disease, 

severity of liver disease and size and location of HCC. 

Complete ablation was defined as the absence of residual disease 

at 3 months post RFA. Recurrence of HCC was defined as 

tumor foci developing at a location of prior RFA treatment – this 

may be early (1 year). Other treatment 

modalities offered to patients were noted. Outcome measures 

included transplant-free survival and tumor recurrence. Results: 

256 patients underwent RFA during the study period. 87% of 

the patients had documented evidence of cirrhosis. 81 patients 

with HBV (79% on treatment), 112 patients with HCV (54% 

treated, 20% SVR), and 63 patients with nonviral etiologies 

were included. 24% of the cohort was female. The mean age 

at diagnosis of HCC was 62.3 (±10.1) years. The mean size 

of the largest lesion at diagnosis was 2.8cm (±1.6cm). All 

patients underwent at least one session of RFA (mean 1.9 treatments). 

The average size of RFA-treated lesions was 2.4cm 

(±0.96cm). 79% of the patients had complete ablation. Recurrence 

occurred in 74 patients (29%), with mean recurrence-free 

interval of 498 days (±383d). 109 (42%) patients died or 

underwent liver transplant during the follow-up period. The 

primary outcome (transplant or death) was reached in 27% 

of HBV patients (median TFS 1.5 years), 53% of HCV patients 

(median TFS 1.7 years) (p=0.0003 HBV vs. HCV), and 40% 

of nonviral patients (median TFS 1.5 years). In a multivariable 

analysis including age, etiology and MELD score, patients 

with HCV had significantly worse survival than patients with 

HBV (HR 2.6, p


175 

Transcriptomic Analysis Of Human Hepatocytes In 

3-Dimensional Cultures With Maintained Perfusion And 

Transport Offers Insights Into The Pharmacotoxicology 

Of Clinically Relevant Concentrations Of Obeticholic 

Acid. 

Arun J. Sanyal 2 , Robert Figler 1 , Brett R. Blackman 1 , Svetlana 

Marukian 1 , Sol Collado 1 , Mark Lawson 1 , Aaron J. Mackey 1 , 

David Manka 1 , Brian R. Wamhoff 1 , Ajit Dash 1 ; 1 HemoShear Therapeutics, 

Charlottesville, VA; 2 Virginia Commonwealth University, 

Richmond, VA 

NASH has emerged as the most common cause of chronic liver 

disease in the Western world. Recently, obeticholic acid (OCA) 

a semi-synthetic bile acid derivative and farnesoid-X-receptor 

(FXR) agonist has been shown to improve insulin sensitivity in 

humans with type 2 diabetes and liver histology in subjects 

with NASH. The precise mechanisms by which these effects 

are mediated are not fully known. AIMS: To use an unbiased 

transcriptomic analysis of hepatocytes exposed to clinically relevant 

doses of OCA to evaluate its effects on pathophysiologically 

relevant pathways. METHODS: We previously described 

a dynamic 3-dimensional system using liver-derived blood flow 

and transport parameters to restore primary human hepatocyte 

biology, allowing for culture at close to physiological insulin/ 

glucose conditions and eliciting drug responses at clinically-relevant 

concentrations. This system represents an advance over 

existing in vitro systems, which require supra-physiological insulin 

concentrations and are associated with loss of polarity and 

transport functions. We applied this system to gain insights into 

the pharmacotoxicological effects of OCA. Primary hepatocytes 

from 5 human donors were exposed to OCA for 48 hours 

at concentrations approximating clinical therapeutic (0.5 μM) 

and supratherapeutic (10 μM) levels. Whole genome transcriptomics 

was performed by RNAseq, and the data analyzed 

using both unbiased algorithms and biased interrogation of 

specific pathways. RESULTS: A dose-dependent suppression 

of Cyp7a and Cyp27a along with upregulation of bile salt 

canalicular export transporters ABCB4 and ABCB11 and basolateral 

transporters OSTA and OSTB were noted as expected. 

Interestingly, FGF19, not known to be highly expressed in 

hepatocytes, was strongly upregulated in our system by OCA 

suggesting an autocrine suppression of FXR expression. A concomitant 

suppression of LXR and HNF4α signaling associated 

with NROB2 activation was also seen. Pleiotropic effects of 

OCA included suppression of TGFβ as well as TNFα signaling 

pathways, and could explain its beneficial effect of reducing 

inflammatory and fibrotic changes in NASH. Consistent with 

the hypercholesterolemia reported with OCA treatment was the 

upregulation of ApoB, the major lipoprotein of LDL. CONCLU- 

SIONS: OCA, in clinically relevant conditions, induces metabolic, 

anti-inflammatory and anti-fibrotic/oncogenic signaling 

in primary hepatocytes. These findings provide a mechanistic 

basis for OCA effects in NASH. 

Disclosures: 






Robert Figler - Employment: HemoShear Therapeutics, LLC 

Brett R. Blackman - Board Membership: HemoShear LLC; Management Position: 

HemoShear LLC; Patent Held/Filed: HemoShear LLC; Stock Shareholder: 

HemoShear LLC 

Svetlana Marukian - Employment: HemoShear LLC 

Mark Lawson - Employment: Hemoshear 

Aaron J. Mackey - Employment: HemoShear, LLC 

David Manka - Employment: HemoShear Therapeutics; Stock Shareholder: 

HemoShear Therapeutics 

Brian R. Wamhoff - Stock Shareholder: HemoShear, LLC 

Ajit Dash - Employment: HemoShear LLC 

The following authors have nothing to disclose: Sol Collado 

176 

Inhibition of NF-kB by deoxycholic acid induces miR- 

21/PDCD4-dependent hepatocelular apoptosis 

Pedro M. Rodrigues 2 , Marta B. Afonso 2 , André L. Simão 2 , Pedro 

M. Borralho 1 , Cecília M. Rodrigues 1 , Rui E. Castro 1 ; 1 iMed. 

ULisboa & Dep. of Biochemistry and Human Biology, Faculty of 

Pharmacy, University of Lisbon, Lisboa, Portugal; 2 iMed.ULisboa, 

Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal 

MicroRNAs (miRNAs/miRs) play a key regulatory role in 

metabolic liver function. In particular, miR-21 deregulated 

expression has been associated with a wide spectrum of liver 

disorders, contributing to disease development and progression. 

We have demonstrated that deoxycholic acid (DCA), a 

cytotoxic bile acid implicated in the pathogenesis of non-alcoholic 

fatty liver disease, inhibits miR-21 expression in hepatocytes. 

Still, the regulatory mechanisms behind miR-21 inhibition 

and its contribution for cell demise are lacking. We aimed to 

unveil the mechanisms underlying modulation of miR-21 by 

DCA and to evaluate their exact contribution to DCA-induced 

cell death. Primary rat hepatocytes were treated with 25-200 

mM DCA for 24 h. Cell death, viability and caspase-3 activity 

were measured by the ApoTox-Glo Triplex Assay. miR-21 

expression was measured by qRT-PCR. Programmed cell death 

4 (PDCD4), a miR-21 pro-apoptotic target, was evaluated by 

immunoblotting and after transfecting cells with a reporter luciferase 

plasmid. NF-kB, IkB and active caspase-2 levels were 

also measured by immunoblotting, while NF-kB activation was 

evaluated using a specific luciferase assay and by analyzing 

NF-kB subcellular localization. Reactive oxygen species (ROS) 

levels were analysed using the fluorescent probe 2’,7’-dichlorodihydrofluorescein 

diacetate. Finally, for functional studies, 

miR-21, PDCD4, caspase-2 and NF-kB were modulated using 

both genetic and pharmacologic approaches, and a well-established 

antioxidant, N-acetyl-L-cysteine (NAC). Our results 

show that DCA inhibits miR-21 expression in a dose-dependent 

manner, while increasing PDCD4 protein levels, with a 

concomitant decrease in cell viability and an increase in cell 

death, caspase-2/-3 activation and ROS production. Either 

miR-21 overexpression or PDCD4 silencing hampered DCA-induced 

cell death. Furthermore, NF-kB activity was decreased in 

a similar pattern to miR-21 expression in DCA-treated hepatocytes. 

In fact, NF-kB inhibition, using a selective chemical inhibitor 

(BAY 11-7085), further decreased miR-21 and increased 

PDCD4 expression levels and apoptosis by DCA. In agreement, 

opposite results were observed in cells overexpressing 

NF-kB or incubated with NAC. In conclusion, NF-kB represents 

a major target of DCA in regulating the miR-21/PDCD4 pathway 

whereas, in turn, oxidative stress and caspase-2 activation 

are two key upstream mechanisms leading to inhibition of 

NF-kB transcriptional activity by DCA. As such, these signaling 

circuits constitute appealing targets for bile acid- and/or 

miR-21-associated liver pathologies. 

Disclosures: 

The following authors have nothing to disclose: Pedro M. Rodrigues, Marta B. 

Afonso, André L. Simão, Pedro M. Borralho, Cecília M. Rodrigues, Rui E. Castro


177 

Critical role of sirtuin 1-mitofusin 2 axis in ischemia/ 

reperfusion injury in aged livers 

Sooyeon Lee, Kristina L. Go, Rebecca Y. U, Joseph A. Flores-Toro, 

Ivan Zendejas, Kevin E. Behrns, Jae-Sung Kim; Surgery, University 

of Florida, Gainesville, FL 

INTRODUCTION: Ischemia/reperfusion (I/R) injury is a fundamental 

obstacle in liver resection and transplantation surgery. 

Aged livers have markedly less reparative capacity after I/R 

than young livers. Impaired autophagy and consequent onset 

of mitochondrial dysfunction contributes to this age-dependent 

increase in I/R injury. Autophagy, a primary catabolic pathway 

in the liver, is strongly modulated by sirtuin 1 (SIRT1), 

a deacetylase boosting cell survival and longevity. The AIM 

of this study is to investigate the role of SIRT1 in I/R injury in 

aged livers. METHODS: Hepatocytes from male C57/BL6 mice 

at 3 (young) and 26 months (aged) of age were subjected to 

simulated I/R. Biochemical, genetic and imaging analysis were 

performed to assess cell death, autophagy flux, and mitochondrial 

function. RESULTS: While 2 h of ischemia has no effects 

on young hepatocytes, this short-term I/R impaired autophagy, 

and rapidly induced the mitochondrial permeability transition 

and necrotic cell death in aged hepatocytes. Immunoblotting 

analysis indicated a near-complete loss of SIRT1 in aged cells 

after I/R, which did not occur in young cells. Interestingly, 

adenoviral overexpression SIRT1 in aged cells failed to mitigate 

I/R injury, suggesting that additional factor(s) is required 

to support cell survival. Immunoprecipitation approaches 

revealed an interaction of SIRT1 with mitofusin 2 (MFN2), a 

mitochondrial outer membrane protein. MFN2 levels in aged 

cells became undetectable after a short-term I/R, while the 

depletion of MFN2 was not observed in young cells. Similar 

to SIRT1, MFN2 overexpression alone did not confer cytoprotection. 

However, co-overexpression of both MFN2 and SIRT1 

protected aged cells against I/R, implying a critical cross-talk 

between SIRT1 and MFN2. To further investigate the importance 

of SIRT1-MFN2 interaction, deletion mutants of MFN2 

were constructed and expressed into HEK293T cells. Consistent 

with the findings in hepatocytes, SIRT1 overexpression in 

wild type cells not only deacetylated MFN2 but also increased 

autophagic flux. However, autophagy stimulated by SIRT1 was 

subdued in the N-terminal deletion mutants (Δ2-92 and Δ262- 

392), suggesting a pivotal role of the N-terminal domain of 

MFN2 in SIRT1-mediated autophagy induction. CONCLUSION: 

Our findings show that the depletion of both SIRT1 and MFN2 

attributes to defective autophagy, mitochondrial dysfunction 

and cell death in aged hepatocytes after I/R. The SIRT1-MFN2 

axis may be an integral factor that governs survivability of 

aged livers after reperfusion. 

Disclosures: 

The following authors have nothing to disclose: Sooyeon Lee, Kristina L. Go, 

Rebecca Y. U, Joseph A. Flores-Toro, Ivan Zendejas, Kevin E. Behrns, Jae-Sung 

Kim 

178 

HMGB1-driven Feedforward Hepatocyte Necroptosis 

Circuit in Lethal Acetaminophen-induced liver injury. 

Charlotte Minsart 1 , Claire Liefferinckx 1 , Sandrine Rorive 2,3 , Arnaud 

Lemmers 4,1 , Eric Quertinmont 1 , Eric Trépo 1,4 , Isabelle Salmon 2,3 , 

Jacques Devière 4,1 , Christophe Moreno 4,1 , Isabelle A. Leclercq 5 , 

Richard Moreau 6,7 , Thierry Gustot 4,1 ; 1 Laboratory of Experimental 

Gastroenterology, Université Libre de Bruxelles, Brussels, Belgium; 

2 Pathology, Erasme Hospital, Brussels, Belgium; 3 DIAPATH- center 

for microscopy and molecular imaging (CMMI), Gosselies, 

Belgium; 4 Gastroenterology and Hepato-Pancreatology, Erasme 

Hospital, Brussels, Belgium; 5 Laboratory of Hepato-Gastroenterology, 

Institut de Recherche Expérimentale et Clinique, Université 

Catholique de Louvain, Brussels, Belgium; 6 INSERM Unité 1149, 

Centre de Recherche sur l’inflammation (CRI), Paris, France; 7 UMR 

S_1149, Université Paris Diderot, Paris, France 

Background & Aims: Release of damage-associated molecular 

patterns, in particular High-mobility group box (HMGB) 

1, contributes to acetaminophen (APAP)-induced liver injury 

but the mechanisms involved are currently incompletely understood. 

The aim of the study is to investigate the contribution of 

HMGB1 in vivo and in vitro at early time points of the APAP-induced 

liver injury and its role in the propagation of necrosis 

process. Methods: APAP hepatotoxicity was induced in vivo 

by intraperitoneal injection in C57Bl/6 mice and in vitro on 

cultured HepaRG cells. HMGB1 was quantified by ELISA or 

immuno-staining. Cell death was determined by MTT, ALT, LDH 

and caspase-3 assays. Glycyrrhizin (GL) and ethyly pyruvate 

(EP) was used to inhibit HMGB1. Liposomal clodronate was 

administrated to mice to deplete Kupffer cells (KC). Expression 

of HMGB1 receptors was assessed by RT-PCR and flow 

cytometry. Dabrafenib and necrostatin-1was used to inhibit 

receptor-interacting protein (RIP)3 and RIP1 respectively. 

Results: In APAP-challenged mice, GL inhibited the HMGB1 

release (decrease of serum levels and increase in hepatocellular 

retention in centrolobular area) with improved survival. 

Depletion of KC in mice exacerbated APAP-induced hepatocyte 

necrosis and HMGB1 release suggesting that HMGB1 did 

not act through KC activation. Addition of APAP on cultured 

HepaRG induced cell necrosis characterized by LDH release 

without caspase-3 activation, and HMGB1 release. Moreover, 

HepaRG were exposed to APAP for 6 hours and the so-conditioned 

medium induced cell death of unexposed HepaRG. 

Inhibition of HMGB1 by GL or EP reduced APAP- and conditioned 

medium-induced HepaRG necrosis and further HMGB1 

release. Exposure of HepaRG and primary human hepatocytes 

to rhHMGB1 resulted in their death, underlining that HMGB1 

acts directly on hepatocytes. HepaRG expressed previously 

described HMGB1 receptors (TLR2, TLR4 and TLR9) at mRNA 

and protein level. Pre-treatment of HepaRG by dabrafenib, a 

specific RIP3 inhibitor, and not by necrostatin-1prevented this 

HMGB1-induced cell death. Conclusion: HMGB1 contributes 

to APAP-induced liver injury through a RIP3-dependent hepatocyte 

necroptosis using a feedforward mechanism. Inhibition 

of HMGB1 at the early phase of APAP-induced liver injury 

improved animal survival by reducing the propagation of this 

regulated hepatocyte necrosis. 

Disclosures: 

Christophe Moreno - Consulting: Abbvie, Janssen, Gilead, BMS; Grant/Research 

Support: Janssen, Gilead, Roche, Astellas 

Isabelle A. Leclercq - Independent Contractor: Genfit 

The following authors have nothing to disclose: Charlotte Minsart, Claire Liefferinckx, 

Sandrine Rorive, Arnaud Lemmers, Eric Quertinmont, Eric Trépo, Isabelle 

Salmon, Jacques Devière, Richard Moreau, Thierry Gustot


179 

Oxaloacetic Acid Protects the Liver From Warm Ischemia/Reperfusion 

Injury 

Grégory Merlen, Benoit Lacoste, Benoît Dupont, Valérie-Ann Raymond, 

Marc Bilodeau; CRCHUM, Montreal, QC, Canada 

Background: Liver ischemia/reperfusion (I/R) is an important 

cause of liver damage early after liver transplantation, post 

liver resection or during hemorrhagic shock. One of the mechanisms 

of cell death occurring in that setting is the disruption of 

mitochondrial activity that lead to alterations in cellular energy 

metabolism. We hypothesized that oxaloacetic acid (OAA), 

which is at the crossroads of gluconeogenesis, amino acid 

metabolism and the citric acid cycle, could refuel the energy 

metabolism during I/R and therefore protect the liver against 

injury. In vitro, we have already demonstrated that OAA was 

the most potent citric acid intermediate in its capacity to protect 

rat hepatocytes from hypoxia. We have also shown that the 

administration of OAA considerably reduces the extent of liver 

injury in the left portal vein ligation model of warm liver ischemia 

in the rat. We here analyze the potential protective effect 

of this compound in a model of hepatic I/R. Methods: Animals 

were subjected to 1 hour of left portal pedicle ligation (common 

bile duct, left hepatic artery and left portal vein) followed 

by reperfusion by unclamping. Animals treated with OAA 

[100mg/kg] received a bolus injection through the ileocolic 

vein 30 minutes before surgery. The extent of liver I/R injury 

was assessed by serum transaminase levels measurements, histological 

signs of tissue damage and liver weight (edema). 

Results: Treatment with OAA before reperfusion significantly 

decreased the AST levels released in blood. After 2 hours of 

reperfusion the AST and ALT levels were respectively decreased 

by 43%±14% (p


Here, we investigated the underlying mechanisms of immune 

tolerance by the gut-liver axis, focusing on interactions between 

gut microbiota and hepatic immune competent cells. METHODS: 

Male C57BL/6 mice were administered an initial and subsequent 

sub-lethal dose of ConA to induce immunological tolerance. 

Liver mononuclear cells were separated 12 h after the 

final ConA injection and analyzed by flow cytometry. Immune 

cell subset cytokine production stimulated with TLR ligands was 

measured and the composition of intestinal bacterial flora was 

evaluated by T-RFLP and comprehensive metagenomics. Intestinal 

permeability was measured by FITC-dextran following 

ConA injection. Mice were treated with antibiotics (ampicillin, 

neomycin, metronidazole, and vancomycin) or fecal microbiota 

transplantation to clarify the role of the gut-liver axis in liver 

tolerance. RESULTS: A single ConA injection induced severe 

liver inflammation but ConA administration 7 days after initial 

injection increased immunosuppresive CD11c + DCs (D7-cDC) 

and regulatory T cells in the liver and promoted immunological 

tolerance. D7-cDC had regulatory characteristics and produced 

IL-10 and TGF-β upon TLR9 ligand stimulation especially 

from damaged hepatocytes. As an initiation of liver tolerance, 

intestinal permeability was significantly increased at 4 h following 

a single ConA injection. Importantly, the composition of 

intestinal bacterial flora changed and the ratio of Clostridium 

subcluster XIV to Bacteroides increased thereafter. Transplantation 

of fecal microbiota derived from mice post-ConA administration, 

but not from untreated mice, to gut sterilized mice 

induced immunosuppressive CD11c + cDCs and regulatory T 

cells in the liver and reduced liver injury by ConA. A similar 

result was observed in germ-free and gnotobiotic mice inoculated 

with fecal microbiota derived from ConA-injected SPF 

mice. CONCLUSIONS: Dysbiosis with increased intestinal permeability 

following ConA administration promotes the migration 

of immunosuppressive cells in the liver in preparation for 

further liver injury. This study identifies a novel homeostasis 

pathway that regulates immune activation and tolerance in the 

liver through the gut-liver axis. 

Disclosures: 

Takanori Kanai - Grant/Research Support: Mitsubishi Tanabe Pharma Corporation 

The following authors have nothing to disclose: Nobuhiro Nakamoto, Hirotoshi 

Ebinuma, Po-sung Chu, Nobuhito Taniki, Takeru Amiya, Akihiro Yamaguchi, 

Shunsuke Shiba, Hidetsugu Saito 

182 

Chronic ethanol feeding suppresses Con A induced T cell 

response and hepatitis in ALDH2-deficient mice 

Yanhang Gao 1,2 , Yong He 1 , Dechun Feng 1 , Zhou Zhou 1 , Bin 

Gao 1 ; 1 NIAAA, National Institutes of Health, Rockville, MD; 

2 Hepatology, The first hospital of Jilin University, Changchun, 

China 

Background and Aims: Aldehyde dehydrogenase 2 (ALDH2) 

is well known about its role on detoxifying aldehydes in ethanol 

metabolism. An ALDH2 inactivating mutation is the most 

common single point mutation in humans, mostly found in East 

Asians, which can cause acetaldehyde accumulation after 

alcohol consumption. However, how acetaldehyde accumulation 

affects T cell response and hepatitis in ALDH2-deficent 

individuals remains unknown. Methods: Wide-type and ALDH2 

knockout mice were subjected to ethanol feeding for 6 weeks, 

followed by injection of a single dose of Concanavalin A (Con 

A). Liver injury and serum cytokine levels were evaluated. 

Results: Con A injection rapidly induced T cell hepatitis, which 

recapitulates the histological and pathological sequelae of T 

cell-mediated hepatitis in viral hepatitis patients. Compared 

with ethanol-fed wild-type mice, ethanol-fed ALDH2 -/- mice had 

lower degree of liver damage post Con A injection, as demonstrated 

by the lower level of serum alanine aminotransferase 

(ALT), the less infiltration of neutrophils, the fewer number of 

activated macrophages, and the smaller area of necrosis in 

the liver. Furthermore, serum levels of several pro-inflammatory 

cytokines including IFN-γ, TNF-α, MCP-1, IL-4, IL-6, IL-10, 

IL12p70, were lower in ethanol-fed ALDH2 knockout mice than 

in wide-type mice post Con A injection. In agree with serum 

cytokine levels, hepatic activation of the IFN-, IL-6, IL-4 downstream 

signaling molecule signal transducer and activator of 

transcription (STAT1, 3, 6) were lower in ALDH2 knockout mice 

than in wide-type mice. Furthermore, ethanol-fed ALDH2 knockout 

mice had much higher levels of plasma corticosterone than 

ethanol-fed wild-type mice. Inhibition of endogenous glucocorticoid 

activity by pretreatment with the glucocorticoid receptor 

antagonist RU-486 restored Con A-mediated liver injury in 

ALDH2-deficeint mice. Conclusions: Chronic ethanol feeding 

results in greater elevation of plasma corticosterone levels in 

ALDH2 knockout mice than in wild-type mice. These elevated 

corticosterone levels inhibit Con A-induced T cell response and 

hepatitis in ALDH2 knockout mice. ALDH2-deficient individuals 

who are excessive drinkers may have reduced T cell response 

and are more susceptible to hepatitis viral infection. 

Disclosures: 

The following authors have nothing to disclose: Yanhang Gao, Yong He, Dechun 

Feng, Zhou Zhou, Bin Gao 

183 

The antifibrotic effect of IL-4Rα signaling depends on 

macrophage subsets prevalent during liver fibrosis progression 

and reversal 

Shih-Yen Weng 1 , Santosh Vijayan 1 , Xiaoyu Wang 1 , Yilang Tang 4 , 

Kornelius Padberg 1 , Yong Ook Kim 1 , Brombacher Frank 5 , Jeff R. 

Crosby 3 , Michael L. McCaleb 3 , Ari Waisman 4 , Ernesto Bockamp 1 , 

Detlef Schuppan 1,2 ; 1 Institute of Translational Immunology, University 

Medicine, Johannes Gutenberg University, Mainz, Germany, 

Mainz, Germany; 2 Beth Israel Deaconess Medical Center, Boston, 

MA; 3 Isis Pharmaceuticals, Carlsbad, CA; 4 Institute for Molecular 

Medicine, Mainz, Germany; 5 Institute of Infectious Disease and 

Molecular Medicine, Cape town, South Africa 

Background and aims: In response to various stimuli, macrophages 

can be functionally divided into M1 (inflammatory) and 

M2 (anti-inflammatory) subsets. Alteration of the M1-M2 ratio 

likely impacts liver fibrosis progression and reversal. IL-4Rα, 

which is activated by IL-4 and IL-13 has been linked to M2 

polarization. However, the functional role of IL-4Rα in liver 

fibrosis progression and reversal remained unclear. Methods: 

IL-4Rα -/- and wild type mice were treated with CCL 4 

via oral 

gavage for 6 weeks. Liver fibrosis reversal was investigated 

after 2 weeks of CCL 4 

withdrawal. Antisense oligonucleotides 

(ASO) were applied intraperitoneally (40mg/kg, 3 times per 

week) during the 6 week s of CCL 4 

treatment or the during 2 

weeks off CCL4. Results: At 6 weeks of CCL 4 

treatment, IL-4Rα -/- 

mice had 25% less fibrosis than their wild type littermates. Flow 

cytometry analysis of the liver of IL-4Rα -/- mice showed less 

inflammation indicated by a reduction of B cells, and CD4 

and CD8 T cells. The proportion of resident M1 macrophages 

was significantly increased and that of proinflammatory monocytic 

Ly6c hi macrophages largely reduced in IL-4Rα -/- livers. 

To further validate IL-4Rα function, mice with cell specific deletion 

of IL-4Rα were generated and subjected to CCL 4 

treatment. 

No overt change of liver fibrosis induction was found 

in T cell-specific knockout mice (IL-4Rα ∆CD4 ). However, after 6 

weeks of CCL4-treatment, fibrosis was significantly attenuated 

in mice with myeloid cell-specific IL-4Rα deletion (IL-4Rα ∆LysM ).


In contrast, IL-4Rα ∆LysM mice displayed retarded fibrosis resolution 

as evidenced by slower clearance of hydroxyproline 

and Sirius-Red stained collagen at 2 weeks off CCL 4 

. This was 

accompanied by a reduction of Ly-6c lo restorative and M2 

macrophages and an increase of M1 macrophages. In a therapeutic 

approach, we applied an ASO targeting IL-4Rα (IL-4Rα 

ASO) to CCL 4 

-treated mice. IL-4Rα ASO strongly attenuated 

fibrosis progression but mitigated fibrosis resolution after CCL 4 

withdrawal. Conclusion: Our studies demonstrate that IL-4Rα 

modulates fibrosis progression and reversal in discordant ways 

through macrophages. During progression, IL-4Rα signaling 

increases inflammation and fibrosis by activating Ly6c hi macrophages; 

during reversal, IL-4Rα potentiates restorative (M2, 

Ly6c lo ) macrophage signaling. Accordingly, IL-4Rα ASO treatment 

attenuates fibrosis progression, but retards reversal. 

Disclosures: 

Jeff R. Crosby - Employment: ISIS Pharmaceuticals 

Michael L. McCaleb - Employment: Isis Pharmaceuticals; Stock Shareholder: Isis 

Pharmaceuticals 

The following authors have nothing to disclose: Shih-Yen Weng, Santosh Vijayan, 

Xiaoyu Wang, Yilang Tang, Kornelius Padberg, Yong Ook Kim, Brombacher 

Frank, Ari Waisman, Ernesto Bockamp, Detlef Schuppan 

184 

Liver sinusoidal endothelial cells induce neutrophil 

extracellular traps in liver sterile inflammation via 

IL-33/ST2 axis 

Hai Huang, Hamza Yazdani, Patricia Loughran, Heth R. Turnquist, 

Allan Tsung; Department of Suegery, University of Pittsburgh Medical 

Center, Pittsburgh, PA 

Both liver sinusoidal endothelial cells (LSECs) and neutrophils 

are involved and interact in liver ischemia/reperfusion (I/R) 

injury. Damaged LSECs, as the major sources of IL-33, play 

important role in neutrophil infiltration during liver I/R. We 

recently found that in response to damage associated molecular 

patterns (DAMPs), infiltrated neutrophils forming neutrophil 

extracellular traps (NETs), exacerbate sterile inflammatory 

injury during liver I/R. We here sought to determine the role 

of IL-33 released from LSECs in NET formation during liver I/R. 

WT or IL-33 KO mice were subjected to a non-lethal warm liver 

I/R model. Recombinant IL-33 (rIL-33) or soluble ST2 (sST2, 

decoy receptor of IL-33) was administered in select groups. 

IL-33 KO mice or sST2-treated WT mice were significantly protected 

from I/R injury, having significantly less serum sALT and 

hepatic necrosis, lower levels of systemic cytokines, abrogation 

of proinflammatory signaling pathways compared to WT mice. 

rIL-33 administered during I/R exacerbated hepatotoxicity and 

systemic inflammation. Although significant neutrophils infiltration 

was found in both IL-33 KO and WT mice, NETs formation 

decreased in IL-33 KO mice during liver I/R compared 

with WT mice, associated with significant less serum level of 

myeloperoxidase (MPO)-DNA complexes and tissue level of 

citrullinated-histone H3 (NET markers). In addition, treatment 

of sST2 reduced NET formation whereas significant increased 

NETs were found in rIL-33 treated mice after liver I/R. In vitro, 

IL-33 released from LSECs that were subjected to hypoxia (1% 

O 2 

) promotes NET formation. Directly using rIL-33 stimulates 

neutrophils confirmed IL-33/ST2-MyD88 signaling pathway in 

NET formation. Using either IL-33 KO LSECs, or co-stimulated 

neutrophil with IL-33 neutralizing antibody or sST2 blocked 

NET formation. Our study demonstrates IL-33 from LSECs initiates 

a feed-forward mechanism to neutrophils inducing NETs 

formation in excessive inflammation and hepatotoxicity during 

liver I/R. 

Disclosures: 

The following authors have nothing to disclose: Hai Huang, Hamza Yazdani, 

Patricia Loughran, Heth R. Turnquist, Allan Tsung 

185 

15-PGDH prevents LPS/GalN-induced acute liver injury 

through inhibiting Kupffer cell activation 

Lu Yao, Chang Han, Tong Wu; pathology and laboratory medicine, 

tulane university, New Orleans, USA Minor Outlying Islands 

The NAD+-dependent 15-hydroxyprostaglandin dehydrogenase 

(15-PGDH) catalyzes the oxidation of the 15(S)-hydroxyl 

group of Prostaglandin E 2 

(PGE 2 

), converting the pro-inflammatory 

PGE 2 

to the anti-inflammatory 15-keto-PGE 2 

(an 

endogenous ligand for peroxisome proliferator-activated receptor-gamma 

[PPAR-γ]). PPAR-γ is a ligand-dependent transcriptional 

factor, which plays an important role in regulation of 

inflammatory cell activation. To evaluate the significance of 

15-PGDH/PPAR-γ cascade in liver inflammation, we generated 

transgenic mice with targeted expression of 15-PGDH in the 

liver (15-PGDH Tg) and the animals were subjected to lipopolysaccharide 

(LPS)/Galactosamine (GalN) induced acute liver 

inflammation and tissue damage. Compared to the wild type 

mice, the 15-PGDH Tg mice showed lower levels of alanine 

aminotransferase (ALT) and aspartate aminotransferase (AST), 

less liver tissue damage, less hepatic apoptosis/necrosis, less 

macrophage activation, and lower inflammatory cytokine production. 

In cultured Kupffer cells, treatment with 15-keto-PGE 2 

or the conditioned medium (CM) from 15-PGDH Tg hepatocyes 

inhibited LPS-induced cytokine production. Both 15-keto-PGE 2 

and the CM from15-PGDH Tg hepatocyes also significantly 

up-regulated the expression of PPAR-γ down-stream genes in 

Kupffer cells. On the other hand, 15-PGDH overexpression 

or 15-keto-PGE 2 

treatment of hepatocytes did not influence 

TNF-α-induced hepatocyte apoptosis. These results suggest 

that the resistance of 15-PGDH Tg mice to LPS/GalN-induced 

liver injury is mediated through 15-keto-PGE 2 

, which activates 

PPAR-γ in Kupffer cells and inhibited inflammatory cytokine 

production. Consistent with this assertion, we observed that the 

PPAR-γ antagonist, GW9662, reversed the effect of 15-keto- 

PGE 2 

in Kupffer cell (in vitro) and restored the susceptibility 

of 15-PGDH Tg mice to LPS/GalN-induced acute liver injury 

(in vivo). Taken together, our findings provide novel evidence 

that hepatic overexpression of 15-PGDH protects against 

LPS/GalN-induced acute liver injury by inhibiting Kupffer cell 

inflammatory response. Thus, 15-keto-PGE 2 

is an endogenous 

PPAR-γ ligand that may be utilized as a pharmacological agent 

to ameliorate liver inflammation and tissue damage.


Disclosures: 

The following authors have nothing to disclose: Lu Yao, Chang Han, Tong Wu 

186 

IL-17 signaling in hepatocellular carcinoma promoted 

by ethanol. 

Hsiao-Yen Ma 2,1 , Jun Xu 1,2 , Mengxi Sun 1,2 , David A. Brenner 2 , 

Tatiana Kisseleva 1 ; 1 Department of Surgery, UC San Diego, La 

Jolla, CA; 2 Department of Medicine, UC San Diego, La Jolla, CA 

Hepatocellular carcinoma (HCC) is a malignant tumor made of 

cells dedifferentiated from mature hepatocytes, which usually 

arises in patients with end stage cirrhosis. Progression of HCC 

is associated with upregulation of inflammation and constitutive 

activation of STAT3. Meanwhile, prolonged alcohol consumption 

has strong immunosuppressive and hepatotoxic effects, 

implying chronic alcohol consumption would promote HCC 

development. Furthermore, ALD patients showed significant 

increase in plasma and hepatic IL-17A expression. Blockage 

of IL-17 signaling significantly reduced alcohol induced steatohepatitis 

and fibrosis in preclinical mouse models. However, 

the role of IL-17 signaling in alcohol promoted HCC remains 

unclear. AIM: To determine the role of IL-17A signaling pathway 

during alcohol promoted HCC development and evaluate 

IL-17A as a therapeutic target for HCC. METHODS: Two different 

models were used to assess the IL-17 signaling in alcohol 

promoted HCC. First, hepatic carcinogen (DEN) induced 

HCC in which WT and IL-17RA null mice (KO) were injected 

with single dose of DEN at 14 days old. Second, steatohepatitis 

associated HCC in which MUP-uPA transgenic mice were 

crossed with WT and IL-17RA KO to generate MUP-uPA|WT 

(MWT) and MUP-uPA|IL-17RA KO (MKO) mice. By 12 weeks 

old, each genotype was assigned to pair-fed group and EtOH 

group to receive Liber-Decarli HFD or alcohol diet for 18 weeks 

respctively. The HCC development in both WT and KO mice 

were compared and Stat3 signal pathway was evaluated. To 

investigate how IL-17A signal regulating HCC associated fibroblasts 

and macrophage, MC-38-GFP cancer cells were injected 

into the spleen of WT and KO recipients for 10 days and the 

tumor size and numbers were compared. RESULTS: Chronic 

administration of the Liber-DeCarli alcohol diet accelerated 

tumorogenesis in both WT/MWT and KO/MKO mice. Blockade 

of IL-17 signaling significantly suppressed tumor size and 

tumor number in both pair-fed and EtOH group. In MUP-uPA 

mice, liver fibrosis was increased after EtOH feeding, which 

was decreased by blockade of IL-17 signaling. Blockade of 

IL-17 signaling showed less inflammation (reduced TNFa, IL1b, 

TGFb1), mediators of metastasis (MMP7, MMP9) and ROS 

production (Nox1, Nox2). As expected, CYP2E1 was elevated 

in EtOH group, but suppressed in KO mice. CONCLUSION: 

Chronic alcohol consumption accelerated tumor development 

which was mediated by IL-17 signaling pathway. The role of 

IL-17 signaling during alcohol promoted HCC may be via regulating 

STAT3 signaling pathway and upregulation of CYP2E1. 

Disclosures: 

The following authors have nothing to disclose: Hsiao-Yen Ma, Jun Xu, Mengxi 

Sun, David A. Brenner, Tatiana Kisseleva 

187 

Deletion of fibrocytes in mice attenuates experimental 

liver fibrosis. 

Jun Xu 1,2 , Min Cong 1,2 , Tae Jun Park 1,2 , David A. Brenner 1 , Tatiana 

Kisseleva 2 ; 1 Department of Medicine, UCSD, San Diego, CA; 

2 Department of Surgery, UC San Diego, La Jolla, CA 

BACKGROUND:Bone marrow (BM) fibrocytes, designated as 

CD45 + and Collagen type I + (Col1a1) cells, are recruited to 

the injured liver, however, their role in liver fibrosis remains 

unclear. AIM: To determine the role of fibrocytes in pathogenesis 

of liver fibrosis. METHODS: 1The contribution of fibrocytes to 

liver fibrosis was studied in BM chimeric mice devoid of fibrocytes 

(DFibrocyte mice), in which fibrocyte death was induced 

by expression of Diphtheria toxin a (DTA) in CD45 + Col1a1 + 

fibrocytes. Specifically, tamoxifen-inducible Col1a1 ER-Cre 

mice were crossed with Rosa26 flox-Stop-flox-YFP reporter mice ± 

Rosa26 flox-Stop-flox-DTA mice, and used as donors for BM transplantation 

into lethally irradiated wt mice to generate wt and 

DFibrocyte mice. 2The role of Col1a1 in regulation of fibrocyte 

function was studied in BM chimeric Col1a1 5’SL-/- -into-wt 

mice, in which mutation of 5’stemloop (5’SL -/- mice) prevented 

proper Col1a1 translation in fibrocytes. 3)All mice were subjected 

to CCl 4 

for 6 w. The therapeutic potential of Serum 

Amyloid P (SAP), a natural inhibitor of fibrocytes, was tested. 

4To translate our findings to humans, the role of fibrocytes 

as prognostic biomarker was evaluated in patients with liver 

cirrhosis. RESULTS: Cell fate mapping revealed that 20% of 

hepatic fibrocytes become a-SMA myofibroblasts, while 80% 

become myeloid cells, which serve as a significant source of 

TGFβ1, IL-6, and IL-1b1 in CCl 4 

treated wt mice. Deletion of 

fibrocytes/progeny in DFibrocyte mice resulted in inhibition 

of CCl 4 

induced liver fibrosis by 50%, as shown by reduced 

Col1a1, a-SMA, and TGFb1 mRNA expression. Interestingly, 

fibrocyte ablation also resulted in suppression of hepatic pro-inflammatory 

macrophages (M1) and promote proliferation of 

anti-inflammatory macrophages (M2), suggesting that fibrocytes 

regulate M1/M2 macrophage polarization. Next, we 

tested if Col1a1 expression affects fibrocyte function. Indeed, 

liver fibrosis was reduced in Col1a1 5’SL-/- into-wt mice by >30%, 

and was associated with impaired proliferation of BM fibrocytes. 

Surprisingly, activation of non-fibrocyte-derived myeloid 

lineages was also reduced, indicating that fibrocytes have an 

important immunoregulatory function. In support, the number 

of circulating fibrocytes in patients correlated with the stage 

of liver fibrosis, and with low levels of serum SAP. Administration 

of SAP significantly ameliorated liver fibrosis in CCl 4 

-wt 

mice. CONCLUSION: Fibrocytes contribute to liver fibrosis indirectly 

by increasing liver inflammation and secreting fibrogenic 

agonists. Targeting fibrocytes with SAP may become a novel 

therapy of liver fibrosis in patients with reduced serum levels 

of SAP. 

Disclosures: 

The following authors have nothing to disclose: Jun Xu, Min Cong, Tae Jun Park, 

David A. Brenner, Tatiana Kisseleva


188 

Splenectomy attenuates advanced liver fibrosis stimulating 

hepatic accumulation of anti-fibrogenic monocytes/ 

macrophages 

Yuji Iimuro 1,2 , Akito Yada 1 , Toshihiro Okada 1 , Naoki Uyama 1 , 

Jiro Fujimoto 1 ; 1 Department of Surgery, Hyogo College of Medicine, 

Hyogo, Japan; 2 Surgery, Nirasaki Municipal Hospital, Nirasaki, 

Japan 

Splenectomy (SP), which is performed in cirrhotic patients with 

hypersplenism, has been reported to improve liver function; 

however the underlying mechanism remains obscure. The AIM 

of the present study was to investigate the mechanism using 

a murine model and human liver tissues. Methods: C57BL/6 

male mice were allowed to drink water including thioacetamide 

(TAA: 300 mg/l) ad libitum for 32 weeks. After SP at 

32 weeks, mice were sacrificed on days 1, 7, and 28, respectively, 

while TAA-administration was continued. Perioperative 

changes in peripheral blood and liver tissues were analyzed. 

In cirrhotic patients with hepatocellular carcinoma (HCC), who 

underwent laparoscopic SP in advance, non-tumorous liver 

tissues obtained from resected specimen were histologically 

analyzed. Results: TAA treatment of mice for 32 weeks reproducibly 

achieved advanced liver fibrosis with splenomegaly, 

thrombocytopenia, and leukocytopenia, well representing 

the compensated liver cirrhosis in humans. After SP, thrombocytopenia 

and leukocytopenia were quickly improved, and 

liver fibrosis was obviously attenuated. Among the leukocytes, 

monocytes/macrophages were selectively increased in peripheral 

blood, as well as in the liver. Meanwhile, progenitor-like 

cells expressing CK-19, EpCAM, or CD-133 appeared along 

the fibrous scar in the liver after TAA treatment, and gradually 

disappeared after SP. Monocytes/macrophages accumulated 

in the liver, most of which were negative for Ly-6C, existed 

adjacent to the hepatic progenitor-like cells. qRT-PCR indicated 

increased canonical Wnt and decreased Notch signals in 

the liver, and Wnt2 protein expression in monocytes/macrophages 

was confirmed. A significant amount of β-catenin accumulated 

in the adjacent progenitor-like cells, and Ki67-positive 

relatively small hepatic cells were significantly increased. Protein 

expression of MMP-9, to which Ly-6G-positive neutrophils 

contributed, was also increased in the liver after SP. In cirrhotic 

patients with HCC, thrombocytopenia and lekocytopenia were 

quickly attenuated after SP, and the percentage of monocytes 

in peripheral blood was selectively increased. In the their liver 

tissues, remarkable accumulation of round- or oval-shaped macrophages/monocytes 

positive for CD163 was detected after 

SP, and they existed closely to the CK19-positive cells, which 

spread out from the ductular reactions, suggesting interaction 

between these cells. Conclusions: The hepatic accumulation of 

monocytes/macrophages, the reduction of fibrosis, and the 

gradual disappearance or expansion of hepatic progenitor-like 

cells, possibly play significant roles in the tissue remodeling 

process in cirrhotic livers after SP. 

Disclosures: 

The following authors have nothing to disclose: Yuji Iimuro, Akito Yada, Toshihiro 

Okada, Naoki Uyama, Jiro Fujimoto 

189 

The role of Mesothelin in the pathogenesis of cholestatic 

liver fibrosis 

Yukinori Koyama 2,1 , Ping Wang 2,1 , David A. Brenner 1 , Tatiana 

Kisseleva 2 ; 1 Medicine, University of Californiam, San Diego, La 

Jolla, CA; 2 Surgery, University of California, San Diego, CA 

Background: There are no curative treatments for patients 

with the cholestatic liver diseases, primary sclerosing cholangitis 

(PSC), and primary biliary cirrhosis (PBC). Although the 

etiology of PSC and PBC remains unclear, activated portal 

fibroblasts (aPFS) have been implicated in pathogenesis of 

cholestatic clinical and experimental liver fibrosis. We have 

identified that mesothelin (Msln) is a specific marker for portal 

fibroblasts. Aim: To examine the role of Msln in cholestatic liver 

fibrosis in mice. Methods: Using Msln as a specific marker of 

aPFs, we investigated the development of BDL-induced liver 

fibrosis in mice devoid of aPFs (Msln DTA mice) that were generated 

by expression of Diphtheria toxin-a (DTA) in Msln + cells 

(by crossing the Msln ER-Cre and Rosa26 flox-Stop-flox-DTA mice). 

Next, the role of Msln in cholestatic liver fibrosis was examined 

in Msln knockout mice (Msln -/- mice). To explore the role Msln 

in TGF-β signaling, we generated transgenic Msln DSmad4 mice 

in which Smad4 was specifically deleted in Msln + aPFs. Wt 

littermates served as controls. All mice were subjected to bile 

duct ligation (BDL), and livers were analysed for development 

of liver fibrosis. aPFs from wt and Msln -/- mice were isolated 

and analysed by RNA-Seq. Finally, to examine if Msln can be 

a new therapeutic target for cholestatic liver fibrosis, BDL wt 

mice were treated with anti-Msln Ab (5 mg, 10 mg / mouse). 

Results: aPF-ablated Msln DTA mice developed 50% less fibrosis 

in response to BDL than wt mice, demonstrating the requirement 

of aPFs for cholestatic liver fibrosis. Msln-/- mice had 

less fibrosis in response to BDL (5 and 17 days), but this effect 

was not observed in carbontetrachloride (CCl 4 

)-treated mice, 

demonstrating a role for Msln in cholestatic but not hepatotoxic 

induced liver fibrosis. Msln -/- aPFs exhibit a defect in migration 

and proliferation by modulating TGF-β1 signaling, as demonstrated 

by reduced number of cells populating the scratch area, 

and decreased expression of Ki-67 by Msln -/- aPFs. Furthermore, 

we found that expression of TGF-β1 target genes, such 

as TGF-β2, PAI-1 and Activin1, were strongly downregulated 

in Msln -/- aPFs compared with wt aPFs, and were associated 

with reduced phosphorylation of Smad2 when stimulated with 

TGF-β1. The Msln DSmad4 mice developed less fibrosis. Administration 

of anti-Msln antibody to BDL-injured mice injury attenuated 

liver fibrosis in a dose dependent manner (compared to 

IgG control). Conclusion: aPFs and in particular the expression 

of Msln in aPFs play important roles in the pathogenesis of 

cholestatic liver fibrosis. Msln might become a novel target for 

treatment of cholestatic liver fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Yukinori Koyama, Ping Wang, 

David A. Brenner, Tatiana Kisseleva


190 

Knockout of the substance P receptor, neurokinin-1, 

reduces liver fibrosis in cholestatic bile duct ligated (BDL) 

mice 

Ying Wan 2 , Nan Wu 3 , Julie Venter 3 , Yuyan Han 3 , Holly A. 

Standeford 4 , Haibo Bai 2 , Shanika Avila 3 , Heather L. Francis 1 , 

Lindsey Kennedy 4 , Micheleine Guerrier 3 , Tina Kyritsi 3 , Tami Annable 

4 , Shannon S. Glaser 1 , Gianfranco Alpini 1 , Fanyin Meng 1 ; 

1 Research, S&W DDRC and Medicine, Central Texas Veterans 

Health Care System and Scott & White, Temple, TX; 2 Operational 

Funds, BaylorScott&White, Temple, TX; 3 Medicine, Texas A&M 

University HSC, Temple, TX; 4 Research, Central Texas Veterans 

Health Care System, Temple, TX 

During the progression of cholestatic liver diseases, proliferating 

cholangiocytes acquire neuroendocrine features and 

secrete sensory neurotransmitters such as α-calcitonin gene 

related peptide (α-CGRP) and substance P (SP). We have previously 

shown that: (i) SP, a proteolytic product of tachykinin 

(Tac1) that is deactivated by membrane metalloendopeptidase 

(MME), stimulates cholangiocarcinoma growth by interacting 

with its receptor, NK1R; and (ii) knockdown of NK1R decreases 

biliary hyperplasia in cholestatic bile duct ligated (BDL) mice. 

Thus, we aim to determine the autocrine/paracrine role of the 

SP/NK1R axis on liver fibrosis in normal and cholestatic mice. 

Methods: In normal and BDL WT and NK1RKO mice, we measured 

serum SP levels and the mRNA expression of Tac1 and 

MME in total liver samples and purified small and large cholangiocytes. 

We measured: (i) liver fibrosis by Sirius Red staining 

in liver sections; and (ii) the mRNA expression of the fibrosis 

genes, α-SMA and fibronectin, in total liver tissues from: (i) 

normal wild-type (WT) mice treated for 1 wk with saline or 

SP (2.5 mmol/Kg BW by IP implanted minipumps); and (ii) 

normal and BDL WT and NK1R KO mice. In vitro, murine 

cholangiocyte lines were treated with BSA (basal) or SP (100 

nM) in the absence/presence of L-733, 060 (25 μM, NK1R 

antagonist) for 6-72 hr before evaluating the expression of the 

aforementioned fibrotic genes. Results: There was an increase 

in SP serum levels in normal WT mice treated with SP and in 

BDL WT compared to normal WT mice. The expression of Tac1 

increased in BDL WT mice, whereas MME mRNA expression 

decreased in BDL WT compared to normal mice; an opposite 

pattern was observed in BDL NK1RKO mice compared to 

BDL WT mice. There was enhanced fibrosis and fibrosis gene 

expression in normal WT mice treated with SP and in BDL WT 

mice compared to normal WT mice. A significant decrease 

in liver fibrosis and the expression of α-SMA and fibronectin 

was observed in BDL NK1RKO mice compared to BDL WT 

mice. Small cholangiocytes isolated from WT BDL mice and 

NK1RKO mouse livers displayed a less fibrotic phenotype 

when compared to large cholangiocytes. In addition, large 

cholangiocytes derived from NK1RKO mice showed a significant 

reduction in fibrotic markers relative to BDL WT mice. 

Treatment of large murine cholangiocytes with SP increased the 

expression of fibrosis genes, whereas L-733, 060 reduced the 

SP-induced expression of α-SMA and fibronectin. Conclusion: 

We propose that modulation of the synthesis of neurotransmitters 

modulated by sensory innervation may be important in the 

modulation of liver fibrosis during the progression of cholestatic 

disorders. 

Disclosures: 

The following authors have nothing to disclose: Ying Wan, Nan Wu, Julie Venter, 

Yuyan Han, Holly A. Standeford, Haibo Bai, Shanika Avila, Heather L. Francis, 

Lindsey Kennedy, Micheleine Guerrier, Tina Kyritsi, Tami Annable, Shannon S. 

Glaser, Gianfranco Alpini, Fanyin Meng 

191 

Loss of Cytoglobin Exacerbates Liver Fibrosis and Cancer 

Development in Steatohepatitis by Activating the 

Oxidative Stress Pathway 

Thuy T. Le 2 , Yoshinari Matsumoto 2 , Tuong Thi Van Thuy 2 , Hoang 

Hai 2 , Katsutoshi Yoshizato 1 , Norifumi Kawada 2 ; 1 Academic Advisor’s 

Office, PhoenixBio Co.Ltd., Hiroshima, Japan; 2 Department 

of Hepatology, Graduate School of Medicine, Osaka City University, 

Osaka, Japan 

Objective: To clarify the role of cytoglobin (CYGB) in the 

development of liver fibrosis and cancer in humans and in a 

mouse model of non-alcoholic steatohepatitis (NASH). Methods: 

CYGB expression was assessed in patients with NASH or 

hepatocellular carcinoma (HCC). Cygb-deficient mice (Cygb / ) 

and primary hepatic stellate cells (HSCs) isolated from Cygb / 

or wild-type (WT) mice were characterized. A mouse NASH 

model was generated in Cygb / and WT mice by feeding a 

CDAA define diet for 8-32 weeks. A subset of mice was treated 

with the antioxidant N-acetylcysteine (NAC). Histopathology 

and gene expression were analyzed. Results: CYGB was 

expressed in HSCs of intact human livers, and its expression 

was reduced in NASH and HCC patients. In Cygb / mice, 67% 

of those aged 1 to 2 years spontaneously exhibited abnormalities 

and cancer development in multiple organs, including 

the liver, lung, lymph nodes and heart, compared with 4.7% 

in WT mice (p


192 

Circulating exosomes from healthy mice attenuate 

hepatic stellate cell activation and are anti-fibrotic in 

vivo 

Li Chen 1 , Ruju Chen 1 , Sherri Kemper 1 , David Brigstock 1,2 ; 1 Clinical 

& Translational Research, Nationwide Childrens Hospital, Columbus, 

OH; 2 Surgery, The Ohio State University, Columbus, OH 

Exosomes are 50-150 nm membranous vesicles that shuttle a 

complex molecular cargo (miRs, mRNAs, proteins) between 

producer and recipient cells resulting in epigenetic regulation 

of cell function. After their release from producer cells, exosomes 

traverse intercellular spaces and may either be taken 

up by neighboring cells or enter body fluids and be dispersed 

systemically. The goal of our work was to determine if hepatic 

stellate cells (HSC) are regulated by circulating exosomes. 

Exosomes were purified from the serum of healthy or fibrotic 

Swiss Webster male mice and tested for their effect on (i) 

activation or fibrogenesis in primary cultures of mouse HSC, 

or (ii) CCl 4 

-induced liver injury in mice. Exosomes (10μg/ml) 

from the serum of healthy mice resulted in decreased CTGF, 

αSMA or collagen α1(I) mRNA after treatment of D9 HSC for 

24hrs (p < 0.01) whereas transcript expression was increased 

or unaffected when the cells were exposed to serum exosomes 

from fibrotic mice. In a 10-day CCl 4 

injury model in 

male BAC reporter transgenic (TG) CTGF-EGFP FX156GSat/ 

Mmucd Swiss Webster mice expressing GFP under the control 

of the CTGF promoter, hepatic GFP or αSMA expression was 

attenuated by i.p. administration every other day (40μg/g) of 

serum exosomes from healthy mice, but not from fibrotic mice 

(p < 0.01). In 5-wk CCl 4 

fibrosis models, i.p. administration of 

serum exosomes (40 μg/g) from healthy mice during the last 2 

wks of CCl 4 

treatment caused a dramatic and dose-dependent 

decrease in hepatic GFP-CTGF production. This was associated 

with decreased expression of CTGF, αSMA or collagen 

mRNA or protein (p < 0.01). The same effects occurred in 

wild type mice. To identify the principal miRs in serum exosomes 

that are differentially expressed between normal versus 

fibrotic mice, miR profiling was performed on serum exosomes 

using a miRnome miR PCR Array. Non-biased ranking of the 

data identified miR-23b, -34c, -106a, -151, -200b, -455, -483 

-532, -653, and -687 as the principal differentially expressed 

miRs and as candidate anti-fibrotics with the ability to attenuate 

expression of CTGF, αSMA or collagen in cultured HSC. These 

studies show that circulating exosomes from healthy individuals 

are instrinsically anti-fibrotic and that this property is attributable, 

at least in part, to specific miR constituents. We propose 

that circulating exosomes offer a novel platform for liver fibrosis 

therapy and that continued interrogation of their complex 

molecular payload will result in the identification of additional 

therapeutic candidates. 

Disclosures: 

David Brigstock - Stock Shareholder: FibroGen 

The following authors have nothing to disclose: Li Chen, Ruju Chen, Sherri Kemper 

193 

Combining high-throughput genome-wide RNAi and 

computational pharmacological screening identifies 

glyburide as a potent therapeutic candidate for liver 

disease due to alpha-1-antitrypsin deficiency 

Yan Wang, Murat C. Cobanoglu, Jie LI, Pamela Hale, Tunda Hidvegi, 

Micheal Ewing, Andrew Chu, Gary A. Silverman, Stephen C. 

Pak, Ivet Bahar, David H. Perlmutter; Departments of Pediatrics and 

Computational Biology, University of Pittsburgh School of Medicine, 


Alpha-1-antitrypsin deficiency (ATD) causes severe liver disease 

by the pathologic accumulation of the misfolded alpha-1-antitrypsin 

Z (ATZ) in hepatocytes. Liver transplantation is the only 

therapy currently available for this liver disease. In this study 

we investigated the possibility of discovering novel therapeutic 

drug candidates by interrogating a C. elegans model of ATD 

with genome-wide RNAi screening and computational (druggene 

interaction) pharmacological strategies. High-content 

genome-wide RNAi screening of our C. elegans model of ATD 

was utilized to identify genes that could modify the proteotoxicity 

of ATZ. Out of more than 16,000 RNAi, the initial screen 

identified 104 genes that are potential positive modifiers of 

ATZ proteotoxicity. These genes were then analyzed as potential 

targets of known drugs using the DrugBank 3.0 computational 

database. One of the drugs identified by this approach 

is glyburide (glibenclamide, GLB), a sulfonylurea drug that has 

been used broadly in clinical medicine as an oral hypoglycemic 

agent. In this study, we show that GLB mediates a marked 

decrease in steady-state levels of misfolded ATZ in a mammalian 

cell line model of ATD and that this effect is dose- and 

time-dependent with an optimal dose of 40 mM. Pulse-chase 

labeling experiments indicate that GLB specifically and selectively 

mediates an increase in the rate of intracellular degradation 

of ATZ. Mechanistic studies suggest that GLB increases 

autophagic flux but does not affect the phosphorylation of 

mTOR substrates and so its effect on the autophagy pathway is 

TOR-independent. As GLB is known to induce cellular calcium 

influx via action on the ATP-sensitive potassium channel, we 

investigated whether GLB could affect the intracellular accumulation 

of ATZ by acting on intracellular calcium signaling 

and found that calcium chelator BAPTA-AM partially blocks the 

GLB-mediated reduction of ATZ levels. Most importantly, GLB 

reduces hepatic ATZ load and hepatic fibrosis when administered 

systemically to the PiZ mouse model of ATD. Preliminary 

studies of GLB analogues suggest that its effects on degradation 

of ATZ can be dissociated from its effects on insulin secretion. 

In conclusion, these results indicate: 1) GLB reduces the hepatic 

proteotoxicity of ATZ accumulation by a calcium-dependent, 

mTOR-independent effect on intracellular degradation; 2) GLB 

(analogues) is a very appealing therapeutic drug candidate for 

ATD because of its wide margin of safety; 3) interrogation of 

a genetically tractable disease model by combining computational 

with high-content automated RNAi screening strategies is 

a valid and powerful drug discovery platform. 

Disclosures: 

The following authors have nothing to disclose: Yan Wang, Murat C. Cobanoglu, 

Jie LI, Pamela Hale, Tunda Hidvegi, Micheal Ewing, Andrew Chu, Gary A. 

Silverman, Stephen C. Pak, Ivet Bahar, David H. Perlmutter


194 

Enteral Obeticholic Acid Prevents Hepatic Cholestasis in 

Total Parenteral Nutrition-Fed Neonatal Pigs 

Douglas Burrin 1 , Yanjun Jiang 1 , Zhengfeng Fang 2 , Barbara Stoll 1 , 

Gregory J. Guthrie 1 , Hongtao Wang 3 , Ignacio R. Ipharraguerre 4,5 , 

Jose J. Pastor 4 ; 1 USDA Children’s Nutrition Research Center, 

Department Pediatrics, Baylor College of Medicine, Houston, 

TX; 2 Animal Nutrition Institute, Sichuan Agricultural University, 

Chengdu, China; 3 Pediatric Gastroenterology, Hepatology, Nutrition, 

Department Pediatrics, Baylor College of Medicine, Houston, 

TX; 4 Lucta SA, Barcelona, Spain; 5 Institute of Human Nutrition and 

Food Science, University of Kiel, Kiel, Germany 

Total parenteral nutrition (TPN) is a vital support for neonatal 

infants with congenital or acquired gastrointestinal (GI) disorders 

and requiring small bowel resection. An adverse outcome 

associated with prolonged TPN use is parenteral nutrition 

associated cholestasis (PNAC). We previously showed that 

enteral chenodeoxycholic acid (CDCA) treatment reduced 

PNAC. We hypothesized that the protective effects of CDCA 

were mediated by modulation of FXR-target genes involved 

in bile acid homeostasis. The aim of the current study was to 

compare the physiological effects of a selective FXR agonist, 

obeticholic acid (OCA) vs CDCA on hepatic bile acid homeostasis 

in TPN-fed piglets. Term, newborn pigs were assigned 

to receive complete TPN (PN), TPN + enteral CDCA (30 mg/ 

kg), or TPN+enteral OCA (0.5, 5, 15 mg/kg) daily for 19 d. 

The daily parenteral lipid was Intralipid given at 10 g/kg. Endpoints 

of PNALD and bile acid homeostasis were measured. 

We found that, compared to PN pigs, treatment with high dose 

of OCA (OCA5 and OCA15), but not CDCA and low dose 

of OCA (OCA0.5), reduced serum PNAC markers including 

bilirubin, gamma-glutamyl transferase (GGT), total bile acid, triglyceride, 

and very-low-density lipoprotein. Compared to PN, 

OCA 5 and 15, but not OCA 0.5 or CDCA, reduced the total 

plasma bile acid concentration by increasing the proportional 

transfer of hepatic bile acid into the gallbladder, suggesting 

increased bile flow. TPN-induced ductopenia as measured by 

percentage of intact bile ducts per portal tract was prevented 

by OCA suggesting preservation of bile ducts. The major bile 

acids in plasma were glyco-conjugated forms of CDCA, hyocholic 

acid and hyodeoxycholic acid. OCA5 and OCA15, but 

not CDCA, suppressed hepatic expression of CYP7A1, while 

CYP27A1 and CYP8B1 mRNA remained unchanged. The bile 

acid detoxification enzyme CYP3A29 mRNA was inhibited by 

both CDCA and OCA treatments. OCA, but not CDCA upregulated 

hepatic mRNA involved in hepatobiliary bile acid and bilirubin 

transport into bile including bile salt export pump (BSEP), 

multidrug resistance protein 1(MDR1), and multidrug resistance 

protein 4 (MRP4). OCA5 and OCA15 induced hepatic and 

ileal FGF19 expression more than CDCA in pigs. We further 

found that OCA5 and OCA15, but not CDCA, inhibited the 

hepatic expression of inflammatory marker interleukin-8. Contrary 

to our previous study, we found that CDCA did not prevent 

PNAC. We suspect that this was due to the higher lipid 

load infused. We conclude that enteral OCA is more effective 

than CDCA in prevention of PNAC via upregulation of FXR-target 

genes involved in preservation of hepatobiliary transporters 

and bile duct function. 

Disclosures: 

The following authors have nothing to disclose: Douglas Burrin, Yanjun Jiang, 

Zhengfeng Fang, Barbara Stoll, Gregory J. Guthrie, Hongtao Wang, Ignacio R. 

Ipharraguerre, Jose J. Pastor 

195 

Biliatresone causes GSH reduction in cholangiocytes, 

leading to cholangiocyte damage, increased monolayer 

permeability, and periductular myofibroblasts: insights 

into the mechanism of biliary atresia 

Orith Waisbourd-Zinman 1 , Hong Koh 2,3 , Pierre-Marrie Lavrut 2,4 , 

Shannon Tsai 2 , John R. Porter 5 , Michael Pack 2 , Rebecca G. 

Wells 2 ; 1 Gastroenterology, Hepatology and Nutrition, Children’s 

Hospital of Philadelpahia, Philadelpahia, PA; 2 Gastroenterology, 

Perelman School of Medicine, University of Pennsylvania, Philadelphia, 

PA; 3 Department of Pediatrcis, Division of Gastroenterology, 

Hepatology and Nutrition, Yonsei University College of Medicine, 

Severance Children’s Hospital, Seoul, Korea (the Republic of); 

4 Pathology, Academic Hospital of Lyon, Lyon, France; 5 Biological 

Sciences, University of the Sciences, Philadelphia, PA 

Background: Biliary atresia (BA) is an acquired fibro-obliterative 

disease of unknown etiology affecting the extrahepatic 

bile ducts of the newborn. We previously reported that the 

novel toxin biliatresone causes selective atresia of the extrahepatic 

biliary tree in zebrafish and microtubule instability and 

lumen obstruction in a cholangiocyte 3D culture model. The 

toxin strongly and spontaneously binds to glutathione (GSH). 

We hypothesized that GSH reduction is responsible for the 

effects of the toxin. Methods: We used several mouse cholangiocyte 

culture systems: 2D culture, 3D spheroid culture, 

and ex vivo culture of neonatal extra-hepatic bile ducts. We 

treated these with biliatresone and compounds that regulate 

GSH, then stained and imaged the cells. We measured 

GSH levels in cholangiocytes treated with the compounds at 

different time points. In order to assess lumen permeability, 

we used a rhodamine efflux assay with live cell imaging of 

cholangiocytes in 3D cultures. Results: Cholangiocytes in 2D 

culture demonstrate a 70% decrease in GSH 1h following 

toxin treatment, with recovery to baseline by 6h. In 3D culture, 

lowering GSH with buthionine sulfoximine (BSO) mimicked 

the effects of biliatresone, with microtubule instability, loss of 

polarity, and lumen obstruction. N-Acetyl-L-cysteine (L-NAC), 

which increases GSH, protected 3D cultures from the effects 

of biliatresone. Neonatal bile ducts cultured ex vivo showed 

patchy disruption of the cholangiocyte monolayer and lumen 

obstruction after treatment with biliatresone or BSO, with 

protection by L-NAC but not the stereoisomer D-NAC. A time 

course study of cholangiocytes in 3D culture treated with biliatresone 

showed that microtubule injury occurred first, within 3 

hours, followed by abnormalities in F-actin, polarity, and tight 

junctions, and finally lumen obstruction. Rhodamine remained 

in the lumens of vehicle-treated cholangiocyte spheroids for up 

to 12 h, but effluxed within 3 h after biliatresone treatment, 

suggesting increased epithelial permeability. In the ex vivo bile 

duct model, staining for a-smooth muscle actin was increased 

in the stroma surrounding the cholangiocyte layer after treatment 

with biliatresone, BSO, or D-NAC plus biliatresone, but 

remained at baseline levels after treatment with L-NAC plus biliatresone. 

Conclusions: Biliatresone rapidly decreases GSH in 

mouse cholangiocyte and bile duct cultures, initially disrupting 

microtubules then compromising cholangiocyte polarity and 

increasing lumen permeability. In vivo, this is a potential cause 

of obstruction and periductular fibrosis and may be important 

to the pathophysiology of BA. 

Disclosures: 

The following authors have nothing to disclose: Orith Waisbourd-Zinman, Hong 

Koh, Pierre-Marrie Lavrut, Shannon Tsai, John R. Porter, Michael Pack, Rebecca 

G. Wells


196 

A functional classification of ABCB4 missense variations 

identified in progressive familial intrahepatic cholestasis 

type 3 

Jean-Louis Delaunay 1 , Anne-Marie Durand-Schneider 1 , Claire Dossier 

1 , Thomas Falguières 1 , Julien Gautherot 1 , Tounsia Aït-Slimane 1 , 

Chantal Housset 1 , Emmanuel Jacquemin 2 , Michèle Maurice 1 ; 

1 UMR_S 938, UPMC & Inserm, Paris Cedex 12, France; 2 Hépatologie 

Pédiatrique, APHP, Hôpital Bicêtre, Kremlin-Bicêtre, France 

Progressive familial intrahepatic cholestasis type 3 (PFIC3) is 

caused by bi-allelic ABCB4 variations. More than 70 % of 

disease-causing ABCB4 variations are missense variations. 

The aims of the study were i) to propose a functional classification 

of ABCB4 missense variations; ii) to test the rescue of 

trafficking-defective mutants by cyclosporins. Methods: Twelve 

single-nucleotide missense variations that were identified in 9 

PFIC3 patients (6 homozygous, 3 composite heterozygous) 

and located in or close to transmembrane domains (F357L, 

S346I, P726L, T775M, Q855L, G954S), first intracellular loop 

(T175A), first nucleotide-binding domain (T424A, N510S, 

I541F, L556R), and linker region (R652G), were reproduced in 

the ABCB4 cDNA. The mutants thus obtained were expressed 

in HepG2 (transiently) and HEK 293 cells (stably), and compared 

with wild type ABCB4, to determine their impact on the 

ABCB4 expression or phosphatidylcholine transport activity, 

measured by a fluorimetric assay. Results: Four mutants were 

either fully (I541F and L556R) or partially (S346I and Q855L) 

retained in the endoplasmic reticulum, in an immature endoglycosidase 

H-sensitive form, of lower molecular weight than 

the mature N-glycosidase F-sensitive form. Phosphatidylcholine 

secretion by these mutants was less than 10 % that of wild 

type, except for Q855L (30 %). Rescue of these trafficking 

defects, i.e. exit from the endoplasmic reticulum and increase 

of the mature form at the bile canaliculi, was obtained by cell 

treatments with cyclosporins A or C, and to a lesser extent 

B, D or H. Four mutations with little or no effect on ABCB4 

expression at the bile canaliculi, caused a significant reduction 

(F357L, T775M and G954S) or absence (P726L) of phosphatidylcholine 

secretion. Two mutants (T424A and N510S) were 

normally processed and expressed at the bile canaliculi but 

their stability, assessed by protein decay, was reduced. We 

found no defect of the T175A mutant, nor of R652G, previously 

described as a polymorphism, which herein co-existed 

with another variant (G954S) on the same allele, in two siblings. 

In patients, the most severe phenotypes appreciated by 

the duration of transplant-free survival, were caused by ABCB4 

variants that were markedly retained in the endoplasmic reticulum, 

and expressed in a homozygous status. In conclusion, 

ABCB4 variations can be classified as follows: nonsense variations 

(I), and on the basis of current findings, missense variations 

that primarily affect the traffic (II), activity (III) or stability 

(IV) of the protein. Among missense variants, those in class II 

cause the most severe clinical phenotypes and can be rescued 

by pharmacological therapies. 

Disclosures: 

The following authors have nothing to disclose: Jean-Louis Delaunay, Anne-Marie 

Durand-Schneider, Claire Dossier, Thomas Falguières, Julien Gautherot, Tounsia 

Aït-Slimane, Chantal Housset, Emmanuel Jacquemin, Michèle Maurice 

197 

Liver-specific deletion of the insulin receptor profoundly 

reduces accumulation and proteotoxicity of misfolded 

α 1 

-antitrypsin Z (ATZ) in a mouse model 

Andrew Chu 1 , Tunda Hidvegi 1 , Souvik Chakraborty 1 , Micheal 

Ewing 1 , Amitava Mukherjee 1 , Patrick Araya 2 , Pamela Hale 1 , 

Christine Dippold 1 , Yan Wang 1 , Jie LI 1 , Evelyn Akpadock 1 , Hou 

Ming Cai 1 , Stephen C. Pak 1 , Donna B. Stolz 2 , Gary A. Silverman 

1 , David H. Perlmutter 1 ; 1 Pediatrics - Gastroenterology, Children’s 

Hospital of Pittsburgh of UPMC; University of Pittsburgh, 

Pittsburgh, PA; 2 University of Pittsburgh, Pittsburgh, PA 

Alpha-1-antitrypsin deficiency (ATD)-associated liver disease 

is an important cause of cirrhosis and hepatocellular carcinoma, 

with liver transplantation currently the only treatment 

for advanced disease. Insulin signaling is a key regulator of 

proteostasis mechanisms, with inhibition of insulin signaling 

linked to increased longevity in animals. The purpose of our 

study was to determine whether abrogation of insulin signaling 

reduces ATZ accumulation and proteotoxicity in a mouse 

model of ATD-associated liver disease. To investigate the effect 

of loss of insulin signaling, we generated double transgenic 

mice (IRZZ) by crossing liver-specific insulin receptor knockout 

mice (LIRKO) to the PiZ mouse model of ATD-associated 

liver disease. Histological analysis of livers from male mice 

at 6 months and 12-14 months demonstrated a 60 percent 

reduction of intracellular ATZ globules by PAS-D staining and 

a 50 percent reduction in hepatic fibrosis by Sirius Red staining 

in IRZZ mice compared with PiZ mice. There was also a 

complete elimination of nodular regeneration in the IRZZ liver. 

This is consistent with results from a C. elegans model of ATD 

in which RNAi corresponding to several intermediates in the 

insulin signaling pathway reduce intracellular ATZ accumulation. 

To determine the mechanism of reduced hepatic ATZ load 

in the IRZZ mice, we carried out pulse-chase radiolabelling on 

isolated hepatocytes and found a significant increase in intracellular 

degradation of ATZ in the IRZZ compared to the PiZ 

hepatocytes. To determine whether specific gene expression 

signatures or signaling pathways were involved in the effect 

of insulin receptor deletion, we used transcriptomic analysis to 

compare hepatic RNA levels in IRZZ to PiZ and control (LIRKO 

and wild-type) mice. Compared to PiZ, the gene expression 

pattern in IRZZ mice showed downregulation of collagen V 

consistent with the marked decrease in Sirius Red staining, 

upregulation of genes associated with hepatocellular proliferation/regeneration 

(prominin 1 and 2), and upregulation of 

genes involved in lysosomal function (lysosomal ATPase, H + 

transporting, V0 subunit D isoform, Atp6v0d2). Furthermore, 

using immunofluorescence for the lysosomal membrane protein 

LAMP2 and transmission electron microscopy, we found a 

marked increase in number of lysosomes in the IRZZ as compared 

to the PiZ liver, suggesting activation of the autophagolysosomal 

system in the IRZZ model. These results indicate that 

the insulin signaling pathway suppresses a key mechanism by 

which the liver attempts to protect itself from misfolded ATZ and 

suggest that activation of the autophagolysosomal system is the 

target of this mechanism. 

Disclosures: 

The following authors have nothing to disclose: Andrew Chu, Tunda Hidvegi, 

Souvik Chakraborty, Micheal Ewing, Amitava Mukherjee, Patrick Araya, Pamela 

Hale, Christine Dippold, Yan Wang, Jie LI, Evelyn Akpadock, Hou Ming Cai, 

Stephen C. Pak, Donna B. Stolz, Gary A. Silverman, David H. Perlmutter


198 

Anti-inflammatory and Antifibrotic Activity of NGM282, 

A Novel Variant of FGF19, in an Mdr2-Deficient Mouse 

Model of Primary Sclerosing Cholangitis 

Lei Ling, Mei Zhou, Hong Yang, Marc Learned, Stephen Rossi, 

Alex M. DePaoli, Hui Tian; NGM Biopharmaceuticals, Inc., South 

San Francisco, CA 

Background and Aims: Primary sclerosing cholangitis (PSC) is 

a chronic inflammatory biliary disease characterized by periductal 

fibrosis and stricture formation. Recent data showed 

decreased expression and intraductular production of FGF19, 

which may worsen pre-existing inflammation and fibrosis. 

NGM282 is an engineered variant of FGF19 that inhibits 

Cyp7a1 but lacks the tumorigenic activity seen with FGF19. 

Mice deficient in phospholipid flippase multidrug resistant 

protein 2 (Mdr2-/-) are a preclinical model for PSC as they 

develop liver histopathology similar to human PSC. We studied 

FGF19 and NGM282 in Mdr2-/- mice to characterize the 

potential biologic activity in PSC. Methods: 12wk old Mdr2-/- 

mice with established hepatobiliary disease received 1 dose of 

adeno-associated virus carrying NGM282, FGF19 or control. 

Serum liver enzymes were analyzed at 4 and 24wks post-dose. 

Liver tissue was collected at 24wks and stained with Hematoxylin 

& Eosin and Sirius Red. Results: Significant decreases in 

alkaline phosphatase (ALP) were seen in male and female mice 

at 4wks with FGF19 (males=349+26 U/L to 107+16 U/L; 

females=565+49 U/L to 98+7 U/L; p


Disclosures: 

Douglas Dieterich - Advisory Committees or Review Panels: Gilead, BMS, Abbvie, 

Janssen, Merck, Achillion 



Aijaz Ahmed - Consulting: Bristol-Myers Squibb, Gilead Sciences Inc., Roche, 

AbbVie, Salix Pharmaceuticals, Janssen pharmaceuticals, Vertex Pharmaceuticals, 

Three Rivers Pharmaceuticals; Grant/Research Support: Gilead Sciences 

Inc. 

The following authors have nothing to disclose: Ryan B. Perumpail, Andy Liu, 

Channa R. Jayasekera, Robert J. Wong 

the 300 mg/kg arm compared to 8 (29%) reinfections in the 

control. Conclusions: The majority of LT recipients have CP B or 

C cirrhosis pre-LT and frequent renal dysfunction early post-LT. 

Despite these medical complexities, preliminary results suggest 

that Civacir is well-tolerated in the peri-transplant setting and 

effective (at the 300 mg/kg dose) in preventing HCV reinfection. 

200 

Prevention of Recurrent Hepatitis C in Liver Transplant 

(LT) Recipients with Civacir ® : Preliminary Results on 

Safety and Efficacy, Including Patients with Renal/Liver 

Dysfunction 

Norah Terrault 1 , Roshan Shrestha 2 , Sanjaya K. Satapathy 3 , Sher 

Linda 4 , Jens Rosenau 5 , Jacqueline G. O’Leary 6 , Thomas D. Schiano 

7 , Lewis W. Teperman 8 , James Spivey 9 , Jeffrey Campsen 10 , 

John M. Vierling 11 , Elizabeth C. Verna 12 , David W. Victor 13 , 

George Therapondos 14 , Kalyan R. Bhamidimarri 15 , Seraphin 

Kuate 16 , Gregory Osgood 16 , Judith Blacklidge 16 , Nicole 

Daelken 16 , Shailesh Chavan 16 ; 1 University of California San Francisco, 

San Francisco, CA; 2 Piedmont Transplant Institute, Atlanta, 

GA; 3 University of Tennessee Health Sciences Center, Memphis, 

TN; 4 University of Southern California, Los Angeles, CA; 5 University 

of Kentucky, Lexington, KY; 6 Baylor University Medical Center, 

Dallas, TX; 7 Mount Sinai Medical Center, New York, NY; 

8 New York University, New York, NY; 9 Emory University, Atlanta, 

GA; 10 University of Utah, Salt Lake City, UT; 11 Baylor College of 

Medicine, Houston, TX; 12 Columbia University, New York, NY; 

13 Houston Methodist Hospital, Houston, TX; 14 Ochsner Clinic 

Foundations, New Orleans, LA; 15 University of Miami, Miami, FL; 

16 Biotest Pharmaceuticals Corporation, Boca Raton, FL 

Background and Aims: Prevention of hepatitis C virus (HCV) 

recurrence post-LT is important as it eliminates the risk of severe 

or progressive disease and reduces the complexity of post-LT 

management. Treatment in the peri-transplant period is especially 

difficult in patients with severe renal impairment, which 

often restricts antiviral therapy (AVT). Safe use of hepatitis B 

immune globulin for the prevention of hepatitis B recurrence 

in post-LT patients is well established with >30 years of experience. 

Civacir ® a hepatitis C immune globulin contains a broad 

variety of antibodies against HCV and may offer a safe prophylactic 

approach in the peri-transplant period. Methods: Study 

988 is an ongoing open-label, randomized phase III clinical 

trial evaluating Civacir 200 or 300 mg/kg vs. standard of care 

(no AVT treatment post-LT) in HCV-infected patients (any genotype) 

awaiting LT. Wait-listed patients receiving pre-LT AVT for 

≤24 weeks and with HCV RNA


patients with end-stage liver disease secondary to chronic HCV 

infection will reach age 60 and require LT. While previous 

studies have evaluated trends in HCV-related LT, none have 

specifically investigated HCV-related LT among the BB cohort 

age > 60. Methods: We utilized the United Network for Organ 

Sharing registry to evaluate the impact of chronic HCV infection 

on long term survival following LT among BB patients age > 60 

from 2003-2012 (MELD era). Overall patient survival following 

LT was analyzed with Kaplan Meier methods and multivariate 

Cox proportional hazards adjusted for age, sex, hepatocellular 

carcinoma, race/ethnicity, year of LT, diabetes mellitus, 

MELD score, albumin, ascites, and hepatic encephalopathy. 

Results: Overall, among patients age > 60, representing 15% 

of patients, survival following LT was significantly lower in the 

HCV BB cohort compared to the non-HCV BB cohort (5-year 

survival: 64.5% vs 77.4%, p


203 

Long-term complete prophylaxis withdrawal in liver-transplanted 

patients for HBV-related cirrhosis is safe 

and frequently associated with spontaneous ANTI-Hbs 

seroconversion 

Ilaria Lenci 1,3 , Daniele Di Paolo 1 , Martina Milana 1 , Francesco 

Santopaolo 1 , Leonardo Baiocchi 1 , Valentina Svicher 2 , Laura Tariciotti 

3 , Giuseppe Tisone 3 , Carlo Federico Perno 2 , Mario Angelico 1 ; 

1 Liver Unit, Tor Vergata University, Rome, Italy; 2 Virology Unit, Tor 

Vergata University, Rome, Italy; 3 Liver Transplant Unit, Tor Vergata 

University, Rome, Italy 

Background and aims. Long-term use of hepatitis-B-virus (HBV) 

specific immunoglobulins (HBIg) and/or nucleoside analogs 

(NA) is recommended to prevent HBV reinfection in HBsAg 

positive liver transplant (LT) recipients. Searching for a more 

rational and tailored approach, we showed (J. Hepatol. 2011; 

55:587-593) that weaning of HBV prophylaxis was feasible 

and safe in a selected group of LT recipients considered at 

“low risk” of HBV recurrence (undetectable HBV-DNA at LT 

and undetectable intra-hepatic total HBV-DNA and ccc-DNA 

5 years after LT), with small percentage of HBV recurrence, in 

the absence of clinically relevant events during the first 2 years 

after HBIg and NA withdrawal. We report here the extended 

follow-up of this selected group of LT recipients, up to 6 years 

(median months 78) after complete prophylaxis withdrawal. 

Methods. The study included 30 LT recipients, all HBeAg negative, 

with HBV genotype D, including 6 with HCV and 7 with 

HDV coinfections, who initially received HBIg and NA (mostly 

lamivudine) for at least 5 years following LT. Sequential HBIg 

and NA withdrawal was performed in two six-month periods 

under strict monitoring of HBV virology, based on monthly 

blood tests and liver biopsies every 6 months for the first two 

years. All patients were then followed with clinical and virological 

test at 3-6 months intervals. Results. Only two patients 

(6.7%) underwent sustained HBV recurrence, with HBsAg and 

HBV-DNA reappearance after 45 and 78 months of prophylaxis 

withdrawal. Both received tenofovir therapy and promptly 

returned HBV-DNA negative and did not experience any clinical 

event. Four patients showed a transient HBsAg positivity (2, 

4, 6 and 45 months after HBIg and NA withdrawal, respectively), 

in one case with transient HBV-DNA detectable, followed 

by later anti-HBs seroconversion. Additional 14 patients 

mounted a measurable anti-HBs titer during the long-term follow 

up, without showing transient HBsAg/HBV-DNA detectability. 

Thus, 28 (93.3%) patients were HBsAg and HBV-DNA negative 

and 18 (60%) were anti-HBs positive at completion of the 

extended follow up. The average lastly detected anti-HBs titer 

was 276 IU/L), with 2 patients showing titers >1000 IU/L. 

Anti-HBs seroconversion occurred after a median of 70 months 

(range: 32-70) after complete prophylaxis withdrawal and was 

unrelated to HCV or HDV coinfections. Conclusion. Complete 

prophylaxis withdrawal is safe in patients liver-transplanted 

for HBV-related disease at low risk of recurrence and in the 

majority of cases is followed by spontaneous anti-HBs seroconversion. 

Disclosures: 

Mario Angelico - Advisory Committees or Review Panels: Gilead, Janssen; 

Grant/Research Support: Roche; Speaking and Teaching: GSK, Roche, Gilead, 

Novartis, BMS, Bayer 

The following authors have nothing to disclose: Ilaria Lenci, Daniele Di Paolo, 

Martina Milana, Francesco Santopaolo, Leonardo Baiocchi, Valentina Svicher, 

Laura Tariciotti, Giuseppe Tisone, Carlo Federico Perno 

204 

Impact of Sofosbuvir-Based Regimens on Renal Function 

in Liver Transplant Recipients: Results of a Multicenter 

Study 

Nabiha Faisal 1 , Eberhard L. Renner 1 , Marc Bilodeau 2 , Bandar 

Aljudaibi 3 , Geri Hirsch 4 , Eric M. Yoshida 5 , Tarana Hussaini 5 , 

Peter Ghali 6 , Stephen E. Congly 7 , Mang M. Ma 8 , Jennifer Leonard 

10 , Curtis Cooper 9 , Kevork M. Peltekian 4 , Nazia Selzner 1 , Les 

Lilly 1 ; 1 Multi organ Transplant, Toronto General Hospital, Toronto, 

ON, Canada; 2 University of Montreal, Montreal, QC, Canada; 

3 London Health Sciences, University of Western Ontario, London, 

ON, Canada; 4 Dalhousie University, Halifax, NF, Canada; 5 University 

of British Columbia, Vancouver, BC, Canada; 6 University of 

McGill, Montreal, QC, Canada; 7 University of Calgary, Calgary, 

AB, Canada; 8 University of Alberta, Edmonton, AB, Canada; 9 University 

of Ottawa, Ottawa, QC, Canada; 10 Memorial University, 

St. John’s, NF, Canada 

BACKGROUND & AIMS: The first nucleotide NS5B polymerase 

inhibitor, sofosbuvir (SOF), is not recommended for patients 

with severe renal impairment due to pharmacokinetic studies 

demonstrating higher serum drug and metabolite levels in such 

patients. However, renal function in cirrhotic and non-cirrhotic 

patients treated with SOF-based regimens is often compromised 

yet poorly assessed. This analysis aims to assess the 

impact of SOF-based regimens on renal function in liver transplant 

(LT) recipients with recurrent HCV, and to assess the role 

of renal function on the efficacy and safety of these regimens. 

METHODS: 165 LT recipients across Canada with HCV recurrence 

were treated with SOF-based regimens from January 

2014 to May 2015. Mean patient age was 58±6.85 years; 

the majority were male, GT1, Caucasian and treatment-experienced. 

One third had aggressive HCV in the graft; 50% had 

F3/4 fibrosis. The majority were on tacrolimus based immunosuppression. 

Median time from LT to treatment was 27 (1-309) 

months. Baseline eGFR was calculated by Modification of Diet 

in Renal Disease formula (MDRD) and patients were stratified 

into 3 groups: eGFR60 ml/min. The primary 

outcome was post treatment changes in renal function from 

baseline. Secondary outcomes included sustained virological 

response at 12 weeks after treatment end (SVR12), anemia 

related adverse events (AEs), need for blood transfusions or 

growth factors, serious AEs, treatment discontinuation due to 

AE, or death. RESULTS: An improvement in post treatment renal 

function was seen in the majority (58%) of patients, mostly in 

the SVR12 group (81% vs. 19%). A decline in renal function 

seen in 22%; more marked in poorest renal function (eGFR 


Marc Bilodeau - Advisory Committees or Review Panels: Verlyx, Bayer, Astellas; 

Consulting: GSK; Grant/Research Support: Merck, Synageva; Speaking and 

Teaching: Merck, Vertex, Abbvie, Aptalis, Roche 

Eric M. Yoshida - Advisory Committees or Review Panels: Hoffman LaRoche, Gilead 

Sciences Inc, Abbvie; Grant/Research Support: Abbvie, Hoffman LaRoche, 

Merck Inc, Vertex Inc, Jannsen Inc, Gilead Sciences Inc, Boeringher Ingleheim 

Inc, Astellas; Speaking and Teaching: Gilead Sciences Inc, Merck Inc 

Curtis Cooper - Advisory Committees or Review Panels: Gilead, Abbvie, MERCK; 

Grant/Research Support: MERCK, Gilead, Abbvie; Speaking and Teaching: 

MERCK, Abbvie, Gilead 

The following authors have nothing to disclose: Nabiha Faisal, Bandar Aljudaibi, 

Geri Hirsch, Tarana Hussaini, Peter Ghali, Stephen E. Congly, Mang M. Ma, 

Jennifer Leonard, Kevork M. Peltekian, Nazia Selzner, Les Lilly 

205 

A Randomized Controlled Trial of Sofosbuvir/GS-5816 

Fixed Dose Combination for 12 Weeks Compared to 

Sofosbuvir with Ribavirin for 12 Weeks in Genotype 2 

HCV Infected Patients: The Phase 3 ASTRAL-2 Study 

Mark S. Sulkowski 1 , Norbert Brau 2 , Eric Lawitz 3 , Mitchell L. Shiffman 

4 , William J. Towner 5 , Peter J. Ruane 6 , Brian Doehle 7 , Jing 

Wang 7 , John McNally 7 , Anu Osinusi 7 , Diana M. Brainard 7 , John 

G. McHutchison 7 , Nancy Reau 8 , Keyur Patel 9 ; 1 Johns Hopkins University, 

Baltimore, MD; 2 James J. Peters VA Medical Center, Bronx, 

NY and Icahn School of Medicine at Mount Sinai, New York, NY; 

3 Texas Liver Institute, University Of Texas Health Science Center, 

San Antonio, TX; 4 Liver Institute of Virginia, Richmond, VA; 5 Kaiser 

Permanente, Los Angeles, CA; 6 Ruane Medical, Los Angeles, CA; 

7 Gilead Sciences, Inc., Foster City, CA; 8 University of Chicago 

Medical Center, Chicago, IL; 9 Duke University School of Medicine, 

Durham, NC 

Introduction: GS-5816 is a pangenotypic HCV NS5A inhibitor 

that has demonstrated high efficacy in genotype 1-6 HCV-infected 

patients when administered for 12 weeks in combination 

with sofosbuvir. The Phase 3 ASTRAL-2 study compared the 

efficacy and safety of treatment with a fixed dose combination 

(FDC) of SOF/GS-5816 for 12 weeks to SOF + RBV for 12 

weeks in treatment-naïve and treatment-experienced genotype 

2 HCV-infected patients, with and without cirrhosis. Methods: 

Enrolled patients were randomized 1:1 to receive either SOF/ 

GS-5816 (400 mg /100 mg daily) FDC for 12 weeks or SOF 

(400mg daily) with RBV (1000-1200mg daily) for 12 weeks. 

Randomization was stratified by prior treatment and cirrhosis 

status. HCV RNA was measured with the CAP/CTM HCV 2.0 

assay with LLOQ =15 IU/mL. The primary endpoint was sustained 

virologic response 12 weeks after treatment (SVR12) 

with a pre-specified non-inferiority margin of 10%. All patients 

have completed treatment and are in follow-up. Results: 266 

patients with genotype 2 HCV infection were randomized and 

treated, 134 with SOF/GS-5816 and 132 with SOF+RBV. 

Overall 59% were male, 88% were white, 38% had IL28B 

CC genotype, 15% had prior treatment failure, and 14% had 

compensated cirrhosis. HCV RNA declined rapidly in both 

treatment groups with > 90% of patients having HCV RNA 

< LLOQ at treatment week 4. The SVR4 rate for the SOF/ 

GS-5816 treated patients is 99% (133/134) and for the 

SOF+RBV treated patients is 96% (127/132). There were no 

patients with relapse in the SOF/GS-5816 group compared 

with 4 patients with relapse in the SOF+RBV treated group 

through posttreatment week 4. Complete SVR12 and resistance 

results will be presented. Two patients discontinued treatment, 

one treated with SOF/GS-5816 discontinued on Day 1 for 

adverse events (anxiety, headache and disturbance in attention) 

and one treated with SOF+RBV did not return for study 

visits after week 10. The most common AEs were fatigue, 

headache, nausea, and insomnia and all were more frequently 

observed in the SOF+RBV treated patients. Two patients in 

each treatment group experienced SAEs, none were considered 

related to study drugs. Hemoglobin declined to < 10g/dL 

in 6 (5%) patients taking SOF+RBV but not in patients taking 

SOF/GS-5816. No other significant laboratory abnormalities 

were observed. Conclusions: Treatment with SOF/GS-5816 

FDC for 12 weeks resulted in high SVR4 rates and was well tolerated 

in treatment-naïve and treatment-experienced genotype 

2 HCV-infected patients with and without cirrhosis. Few (


206 

Daclatasvir plus sofosbuvir with or without ribavirin in 

genotype 3 patients from a large French multicenter 

compassionate use program 

Christophe Hezode 1 , Victor de Ledinghen 2 , Hélène Fontaine 3 , 

Fabien Zoulim 4 , Pascal Lebray 5 , Nathalie Boyer 6 , Dominique 

G. Larrey 7 , Christine Silvain 8 , Danielle Botta-Fridlund 9 , Vincent 

Leroy 10,11 , Marc Bourlière 12 , Louis d’Alteroche 13 , Isabelle 

Fouchard-Hubert 14 , Dominique Guyader 15 , Isabelle Rosa 16 , Eric 

Nguyen-Khac 17 , Vincent Di Martino 18 , Fabrice Carrat 19 , Larysa 

Fedchuk 20 , Raoudha Akremi 20 , Yacia Bennai 20 , Jean-Pierre Bronowicki 

21 ; 1 Department of Hépato-Gastroenterology, CHU Henri 

Mondor, Créteil, France; 2 Liver fibrosis investigational center, Haut- 

Lévêque Hospital, Bordeaux University Hospital, Pessac, France; 

3 Department of Hepatology, Cochin Hospital, APHP, Paris, France; 

4 Department of Hepatology, La Croix-Rousse Hospital, Lyon Civil 

Hospices, Lyon, France; 5 Department of Hepato-Gastroenterology, 

Pitié Salpêtrière Hospital, Paris, France; 6 Department of Hepatology, 

Beaujon Hospital, APHP, Paris, France; 7 Department of 

Hepato-Gastroenterology, Saint-Eloi Hospital, CHU Montpellier, 

Montpellier, France; 8 Department of Hepato-Gastroenterology 

and of nutritional support, epithelial tissues and cytokines laboratory 

EA 4331, CHU Poitiers, Poitiers, France; 9 Department 

of Hepato-Gastroenterology, Conception Hospital, Marseille, 

France; 10 Grenoble University hospital, Hepato-Gastroenterology 

University Clinic, Grenoble, France; 11 Inserm, 823 Unity, Grenoble, 

France; 12 Department of Hepatology and Gastroenterology, 

Saint-Joseph Hospital, Marseille, France; 13 Unit of Hepatology, 

Department of Hepato-Gastroenterology, CHU Trousseau, Tours, 

France; 14 Department of Hepato-Gastroenterology, CHU Angers, 

Angers, France; 15 Department of liver diseases, CHU Rennes, 

Rennes 1 University, Rennes, France; 16 Department of Hepatology 

and Gastroenterology, CHI Créteil, Créteil, France; 17 Department 

of Hepato-Gastroenterology, CHU Amiens Nord, Amiens, France; 

18 Department of Hepatology and of digestive intensive care unit, 

CHRU Jean Minjoz, Besançon, France; 19 Inserm UMR-S 707, 

Pierre-et-Marie-Curie University Paris 6, Paris, France; 20 Bristol-Myers 

Squibb Research and Development, Rueil-Malmaison, France; 

21 INSERM Unité 954, CHU Nancy and Lorraine University, Vandoeuvre-lès-Nancy, 

France 

Background and aims. Treatment options for HCV genotype 3 

(GT3) patients are limited. The combination daclatasvir (DCV) 

and sofosbuvir (SOF) for 12 weeks is associated with high 

SVR 12 

rate in genotype 3 non cirrhotic patients (96%) and a 

lower SVR 12 

rate in cirrhotic (70%). GT3 cirrhotic remain a 

difficult to treat population and may benefit from the addition 

of ribavirin (RBV) or extended treatment duration. This analysis 

reports SVR results from an ongoing multicenter compassionate 

use program (ATU) in France. Methods. Patients with GT3 and 

advanced liver disease from 221 centers have been included 

since March 2014 to October 2014. Transplanted patients 

were excluded from this analysis. All patients received DCV+- 

SOF QD for 12 or 24 weeks, with RBV added at the physician’s 

discretion. We report interim SVR 12 

rates in patients who have 

completed treatment to date and for whom at least one follow-up 

visit was available. Results. 395 patients were enrolled 

with the following characteristics: males (75.5%), median 

age (54.4 years; 27-83), HCV mono-infected (80%), cirrhosis 

(n=322; 81.5%), including Child-Pugh A (85.5%), B (11.5%), 

C (3.0%) and treatment-experienced (n=288; 73.1%). Median 

HCV-RNA was 5.99 (1.20; 7.41) log IU/mL and median platelets 

count was 120.0 x10 9 /L (30-387). 71.6% of patients have 

albumin≥ 35 g/L. RBV was given in 20.5% of patients and 

treatment duration was 24 weeks in 42.0% and 12 weeks in 

17.5%. Data on SVR 12 

is available in 78 patients; SVR 12 

was 

achieved in 90% (70/78). SVR 12 

rates according to treatment 

schedule and fibrosis stage are presented in the table. The 

treatment failures were related to relapse in 6 patients and 

unknown reasons in 2 patients (only D0 and SVR 12 

available). 

No virologic breakthrough was observed. Treatment discontinuations 

were related to: 1 adverse event, 4 deaths not related 

to study drugs, 1 patient’s decision, 1 treatment failure and 4 

for unknown reason. Efficacy and safety data will be updated 

and presented in the 395 patients at the congress. CONCLU- 

SION: DCV+SOF±RBV regimen demonstrated high SVR 12 

rate 

in GT3 patients with advanced liver disease. The optimal treatment 

duration was 24 weeks in cirrhotic patients. The impact of 

RBV will be analyzed in the overall population and presented 

at the meeting. 

Efficacy of DCV+SOF±RBV regimens in GT3 patients 

Disclosures: 

Christophe Hezode - Speaking and Teaching: Roche, BMS, MSD, Janssen, abbvie, 

Gilead 



Hélène Fontaine - Board Membership: Abbvie, Gilead, BMS, Janssen; Independent 

Contractor: gilead, BMS, MSD, Roche, Janssen 

Fabien Zoulim - Advisory Committees or Review Panels: Janssen, Gilead, Novira, 

Abbvie, Tekmyra, Transgene; Consulting: Roche; Grant/Research Support: 

Novartis, Gilead, Scynexis, Roche, Novira, Assemblypharm; Speaking and 

Teaching: Bristol Myers Squibb, Gilead 

Pascal Lebray - Grant/Research Support: Merck, astellas, Biotest, BMS; Speaking 

and Teaching: Janssen, MSD, Gilead 

Nathalie Boyer - Board Membership: MSD, JANSSEN, Gilead, Abbvie; Speaking 

and Teaching: BMS 

Dominique G. Larrey - Advisory Committees or Review Panels: BAYER, SANOFI, 

PFIZER, SERVIER-BIG, AEGERION, MMV, BIAL-QUINTILES, TEVA, ORION, NEG- 

MA-LERADS, ASTELLAS, ASTRAZENECA, DNDI, GSK, J AND J; Board Membership: 

BMS, GILEAD, ABBVIE, BMS, GILEAD, ITREAS, MMS; Grant/Research 

Support: GILEAD, MSD, BMS, ABBVIE, TIBOTEC/JANSSEN, BMS 

Danielle Botta-Fridlund - Consulting: GILEAD, ABBVIE, BMS, MSD 



Marc Bourlière - Advisory Committees or Review Panels: Schering-Plough, 

Bohringer inghelmein, Schering-Plough, Bohringer inghelmein, Transgene; Board 

Membership: Bristol-Myers Squibb, Gilead, Idenix; Consulting: Roche, Novartis, 

Tibotec, Abott, glaxo smith kline, Merck, Bristol-Myers Squibb, Novartis, Tibotec, 

Abott, glaxo smith kline; Speaking and Teaching: Gilead, Roche, Merck, Bristol-Myers 

Squibb 

Isabelle Fouchard-Hubert - Speaking and Teaching: JANSSEN, BMS, GILEAD, 

ABBVIE 

Dominique Guyader - Advisory Committees or Review Panels: ROCHE, GILEAD, 

IRIS, ABBVIE; Board Membership: MERCK; Grant/Research Support: JANSSEN; 

Speaking and Teaching: BMS 

Eric Nguyen-Khac - Speaking and Teaching: Gilead, Abbvie, Janssen, Roche, 

MSD, BMS 




Fabrice Carrat - Advisory Committees or Review Panels: BMS; Grant/Research 

Support: Janssen, Merck, Gilead, BMS, Roche 

Larysa Fedchuk - Employment: Bristol-Myers Squibb 

Raoudha Akremi - Employment: BMS 

Jean-Pierre Bronowicki - Consulting: Merck, Janssen, Boehringer Ingelheim, Gilead, 

BMS, Bayer, Novartis, GSK, Merck, Janssen, Boehringer Ingelheim, Gilead, 

BMS, Bayer, Novartis, GSK, ABBVIE, ABBVIE; Speaking and Teaching: Roche, 

Merck, Janssen, BMS, Bayer, Roche, Merck, Janssen, BMS, Bayer, ABBVIE 

The following authors have nothing to disclose: Christine Silvain, Louis d’Alteroche, 

Isabelle Rosa, Yacia Bennai


207 

Antiviral Therapy for Chronic Hepatitis B (CHB) Reduces 

the Incidence of Hepatocellular Carcinoma (HCC) 

Regardless of Cirrhosis Status: Analysis with Adjustment 

for REACH-B Risk Score 

Derek Lin 2 , Hwai-I Yang 3,4 , Nghia H. Nguyen 12,1 , Joseph K. 

Hoang 1 , Yoona Kim 1 , Vinh D. Vu 1 , An K. Le 1 , Kevin T. Chaung 1,5 , 

Vincent G. Nguyen 1,5 , Huy N. Trinh 6 , Jiayi Li 7 , Jian Q. Zhang 10 , 

Ann W. Hsing 8,9 , Chien-Jen Chen 3,11 , Mindie H. Nguyen 1 ; 1 Division 

of Gastroenterology and Hepatology, Stanford University 

Medical Center, Palo Alto, CA; 2 Department of Medicine, Stanford 

University Medical Center, Palo Alto, CA; 3 Genomics Research 

Center, Academia Sinica, Taipei, Taiwan; 4 Institute of Clinical 

Medicine, National Yang-Ming University, Taipei, Taiwan; 5 Pacific 

Health Foundation, San Jose, CA; 6 San Jose Gastroenterology, 

San Jose, CA; 7 Palo Alto Medical Foundation, Mountain View, 

CA; 8 Cancer Prevention Institute of California, Fremont, CA; 9 Stanford 

School of Medicine, Stanford Cancer Institute, Palo Alto, CA; 

10 Chinese Hospital, San Francisco, CA; 11 Graduate Institute of Epidemiology 

and Preventive Medicine, National Taiwan University, 

Taipei, Taiwan; 12 Department of Medicine, University of California 

San Diego, San Diego, CA 

Objective: Benefits of antiviral therapy on HCC incidence 

remains controversial. We aim to examine the effect of antiviral 

therapy for CHB on HCC incidence after adjustment for 

background risks using a previously validated REACH-B risk 

score. Methods: We examined a cohort of 5,908 CHB patients 

which included primary analysis of 2,255 from a United 

States cohort (973 of these received antiviral therapy) and 

3,653 patients from the previously described community-based 

Taiwanese REVEAL-HBV study (none received antiviral therapy). 

We used Cox proportional hazards models to calculate 

the hazard ratios (HRs) of developing HCC with adjustment 

using the previously validated REACH-B risk score (17-point 

composite score of sex, age, ALT, HBeAg status, and HBV 

DNA). Results: There were a total of 273 incident cases: 12 

of 973 treated subjects and 263 of 4935 untreated subjects. 

There was a significant 77% risk reduction in HCC incidence 

in patients with antiviral treatment compared with those without 

treatment (HR 0.23; 95% CI 0.13-0.43; pULN). Conclusion: In this large and diverse 

cohort of clinic and community participants from two countries, 

antiviral therapy was significantly associated with a reduction 

in HCC risk after adjustment for background risk using a validated 

risk score inclusive of 5 major known predictors of HCC. 

To our knowledge, this is the largest assembled CHB cohort to 

examine the effect of antiviral therapy. 

REACH-B Adjusted HCC Incidence by Treatment 

Disclosures: 

Yoona Kim - Employment: Proteus Digital Health 

Huy N. Trinh - Advisory Committees or Review Panels: BMS, Gilead; Grant/ 

Research Support: BMS, Gilead; Speaking and Teaching: BMS, Gilead; Stock 


Mindie H. Nguyen - Advisory Committees or Review Panels: Bristol-Myers 

Squibb, Bayer AG, Gilead, Novartis, Onyx; Consulting: Gilead Sciences, Inc.; 

Grant/Research Support: Gilead Sciences, Inc., Bristol-Myers Squibb, Novartis 

Pharmaceuticals, Roche Pharma AG, Idenix, Hologic, ISIS 

The following authors have nothing to disclose: Derek Lin, Hwai-I Yang, Nghia H. 

Nguyen, Joseph K. Hoang, Vinh D. Vu, An K. Le, Kevin T. Chaung, Vincent G. 

Nguyen, Jiayi Li, Jian Q. Zhang, Ann W. Hsing, Chien-Jen Chen 

208 

A Single Dose of RG-101, a GalNAc-Conjugated Oligonucleotide 

Targeting miR-122, Results in Undetectable 

HCV RNA Levels in Chronic Hepatitis C Patients at Week 

28 of Follow-up 

Meike H. van der Ree 1 , J. Marleen D. Vree 2 , Femke Stelma 1 , 

Sophie Willemse 1 , Marc van der Valk 1 , Svend Rietdijk 3,1 , Richard 

Molenkamp 4 , Janke Schinkel 4 , Salah Hadi 5 , Marten Harbers 5 , 

Andre van Vliet 5 , Joanna Udo de Haes 5 , Paul Grint 6 , Steven 

Neben 6 , Neil Gibson 6 , Hendrik W. Reesink 1 ; 1 Gastroenterology 

and Hepatology, AMC, Amsterdam, Netherlands; 2 Gastroenterology 

and Hepatology, UMCG, Groningen, Netherlands; 3 Gastroenterology 

and Hepatology, OLVG, Amsterdam, Netherlands; 

4 Medical Microbiology, Clinical Virology Laboratory, AMC, 

Amsterdam, Netherlands; 5 PRA Healthsciences, Zuidlaren, Netherlands; 

6 Regulus Therapeutics, San Diego, USA Minor Outlying 

Islands 

Background: MicroRNA-122 (miR-122) plays a crucial role in 

the hepatitis C virus (HCV) life cycle. Binding of miR-122 to 

the 5’UTR of the HCV genome promotes HCV RNA stability 

and accumulation, protects the HCV genome from degradation 

by cellular exonucleases and prevents induction of an innate 

immune response against the virus. The aim of this study was 

to evaluate the safety and efficacy of a single dose of RG-101, 

a carbohydrate conjugated oligonucleotide that targets miR- 

122 in hepatocytes, in chronic HCV patients infected with 

various HCV genotypes. Methods: In this multicenter phase 

1 study, we included 32 chronic HCV patients with genotype 

1, 3 or 4 infection. The first cohort received a single subcutaneous 

injection of 2 mg/kg RG-101 (n=14) or placebo (n=2), 

the second cohort received a single subcutaneous injection 

of 4 mg/kg (n=14) or placebo (n=2). Both cohorts were followed 

8 weeks after randomization, and patients with >2 log


decrease in HCV RNA level from baseline were included in 

an extended follow-up study in which patients were followed 

until 28 weeks after dosing. Patients were eliminated from the 

extended follow-up study if they had > 1 log increase in HCV 

RNA level from nadir. HCV RNA levels were measured using 

Roche COBAS AmpliPrep/COBAS Taqman HCV v2.0 assay, 

with a reported LLOQ of 15 IU/mL. Results: Thirty-two patients 

infected with HCV genotype 1 (n=16), genotype 3 (n=10) or 

genotype 4 (n=6) were included in the study. Twenty-three 

patients were treatment naïve and 9 patients were virological 

relapsers to prior interferon-based therapy. None of the 

patients had cirrhosis. At baseline, mean HCV RNA levels were 

comparable between RG-101 dosed patients versus placebo 

(6.2 versus 6.4 log 10 IU/mL, p=0.53). The mean viral load 

reduction at week 4 was 4.4 log 10 

IU/mL (range 2.3-5.8) in 

patients dosed with RG-101 (p200,000 IU/mL were randomized 

1:1 to receive either TDF from gestation week 30–32 to 

postpartum week 4 or no treatment, and were followed-up until 

postpartum week 28. All infants received immunoprophylaxis. 

The primary measurement was the MTCT rate, while endpoints 

included TDF safety, maternal HBV DNA reduction at delivery, 

and HBeAg or hepatitis B s antigen loss/seroconversion 

at postpartum week 28. Results: Among the 200 mothers 

enrolled in 5 regions of the country, 180 completed the study. 

At postpartum week 28, the MTCT rate was significantly lower 

in infants from TDF-treated mothers when compared to those 

from non-treated mothers, both on per-protocol analysis (0% 

vs. 6.82%, P = 0.013) and intention-to-treat analysis (5.16% 

vs. 18.0%, P = 0.007). The safety profile was similar between 

groups, with no difference in birth defect rates (2.11% with TDF 

exposure vs. 1.14% without exposure, P = 1.00). HBV DNA 

levels decreased to


Baseline Values 

† When compared respective variables between groups, P > 

0.05 except HBeAg+ at birth (P < 0.001) and detectable HBV 

DNA at birth (P = 0.002). *3 mothers withdrew before the baseline 

visit. 

Disclosures: 

Calvin Q. Pan - Advisory Committees or Review Panels: Gilead; Consulting: 

BMS, Gilead, Abbvie, Janssen ; Grant/Research Support: BMS, Gilead, Merck; 

Speaking and Teaching: BMS, Gilead, Abbvie 

The following authors have nothing to disclose: Zhong-Ping Duan, Erhei Dai, 

Shuqin Zhang, Guo Rong Han, Yuming Wang, Huaihong Zhang, Huaibin Zou, 

Bao Shen Zhu, Wen Jing Zhao, Hong Xiu Jiang 

210 

High Efficacy of Grazoprevir/Elbasvir (GZR/EBR) in HCV 

Genotype 1, 4, and 6-infected Patients With HIV Coinfection: 

SVR24 Data From the Phase 3 C-EDGE Coinfection 

Study 

Jürgen K. Rockstroh 1 , Mark Nelson 2 , Christine Katlama 3 , Jacob P. 

Lalezari 4 , Josep Mallolas 5 , Mark Bloch 6 , Gail Matthews 7 , Michael 

Saag 8 , Philippe J. Zamor 9 , Chloe Orkin 10 , Jacqueline Gress 11 , 

Melissa Shaughnessy 11 , Stephanie Klopfer 11 , Anita Y. Howe 11 , 

Janice Wahl 11 , Bach-Yen T. Nguyen 11 , Eliav Barr 11 , Heather L. 

Platt 11 , Michael Robertson 11 , Mark S. Sulkowski 12 ; 1 Bonn Umiversity, 

Bonn, Germany; 2 Chelsea and Westminster Hospital, London, 

United Kingdom; 3 Université Pierre et Marie Curie, Paris VI and 

Hôpital Pitié-Salpêtrière, Paris, France; 4 Quest Clinical Research, 

San Francisco, CA; 5 Hospital Clinic-University of Barcelona, Barcelona, 

Spain; 6 Holdsworth House Medical Practice, Darlinghurst, 

NSW, Australia; 7 St Vincent’s Hospital, Sydney, NSW, Australia; 

8 University of Alabama at Birmingham, Birmingham, AL; 9 Carolinas 

Medical Center, Charlotte, NC; 10 Royal London Hospital, 

London, United Kingdom; 11 Merck & Co., Inc., Kenilworth, NJ; 

12 Johns Hopkins University School of Medicine, Baltimore, MD 

BACKGROUND: The fixed-dose combination (FDC) of grazoprevir 

(GZR, MK-5172; NS3/4 protease inhibitor) and elbasvir 

(EBR, MK-8742; NS5A inhibitor), an interferon-free, ribavirin-free, 

once-daily tablet has shown robust efficacy and safety 

in diverse populations. C-EDGE Coinfection evaluated GZR/ 

EBR among treatment-naive, HIV/HCV coinfected patients with 

GT1, 4 or 6. METHODS: All patients were on stable antiretroviral 

(ARV) therapy (tenofovir or abacavir, and lamivudine or 

emtricitabine; and either raltegravir, dolutegravir or rilpivirine) 

with CD4 >200 cells/mm 3 and an HIV RNA 500 cells/mm 3 and 

HIV RNA 5× resistance to EBR in vitro (L31M, Y93S). Adverse 

events (AEs) were reported in 72% (157/218) of patients; serious 

AEs occurred in 0.9% (2/218) of patients. Adherence was 

>90% in the total population, including virologic failures. PK 

parameters were similar in patients with, and without SVR24. 

2 patients had transient HIV viremia; both were subsequently 

undetectable without a change in ARV regimen. CONCLUSION: 

A 12-week regimen of GZR/EBR FDC was highly effective with 

a favorable safety profile among HIV/HCV coinfected patients 

with GT1, 4 or 6 infection. SVR24 was high in all patient subgroups, 

including African-Americans and those with cirrhosis. 

Disclosures: 





Mark Nelson - Advisory Committees or Review Panels: Janssen, MSD, BMS, 

ABBVIE, Viiv, Gilead; Consulting: Janssen, MSD, BMS, ABBVIE, Viiv, Gilead; 

Grant/Research Support: Boehringer Ingelheim, Janssen, MSD, BMS, ABBVIE, 

Viiv, Gilead, Roche; Speaking and Teaching: GSK, Janssen, MSD, BMS, Abbott, 

Viiv, Gilead 

Josep Mallolas - Board Membership: Boehringer, Merck, Abbvie, Gilead, Janssen 

Michael Saag - Grant/Research Support: BMS, Gilead, Abbvie, Merck, ViiV, 

Janssen 



Janssen 

Chloe Orkin - Advisory Committees or Review Panels: Viiv, gilead, bms, boehringer 

ingelgeim; Grant/Research Support: johnson and johnson, abbvie 

Jacqueline Gress - Employment: Merck, Merck 

Melissa Shaughnessy - Employment: Merck & Co., Inc.; Stock Shareholder: 

Merck & Co., Inc. 








BMS 

The following authors have nothing to disclose: Christine Katlama, Jacob P. 

Lalezari, Mark Bloch, Gail Matthews, Stephanie Klopfer, Heather L. Platt


211 

Clinical Features and Outcomes of Complementary and 

Alternative Therapy Medicine (CAM)-Induced Acute Liver 

Failure and Injury 

Luke Hillman 1 , Daniel Ganger 1 , Michelle Gottfried 2 , Jorge Rakela 3 , 

Michael L. Schilsky 4 , William M. Lee 5 ; 1 Medicine, Northwestern 

University, Chicago, IL; 2 MUSC, Charleston, SC; 3 Medicine, Mayo 

Clinic, Scottdsdale, AZ; 4 Medicine and Surgery, Yale University, 

New Haven, CT; 5 Internal Medicine, University of Texas Southwestern, 

Dallas, TX 

Background: Increased use of CAM across the United States 

is evidenced by increased expenditures and reported use by 

consumers. While idiosyncratic drug-induced liver injury (DILI) 

from conventional prescription medicines is often severe, DILI 

from CAM is similarly infrequent but causes significant hepatotoxicity. 

AIM: We examined clinical features and outcomes 

among patients with acute liver failure (ALF) and acute liver 

injury (ALI) enrolled in the Acute Liver Failure Study Group 

(ALFSG) database, comparing conventional DILI injury to that 

due to CAMs. Patients and Methods: ALFSG has prospectively 

enrolled 2626 subjects with ALF/ALI since 1998 from U.S. 

academic liver transplant centers through January 8, 2015. 

We compared clinical presentation, course and outcome 

between those with ALF/ALI due to ‘conventional’ DILI versus 

CAMs. Results: We found 251 (10.4%) subjects deemed by 

DILIN criteria to have DILI-ALF, 210 (83.7%) due to conventional 

medicines and 41 (16.3%) from CAM, with a higher % 

of DILI cases due to CAM between 2008-2015, as compared 

to 1998-2007 (18.1% vs. 12.4%, p = 0.047). There was no 

difference in the type of liver injury (hepatocellular, cholestatic, 

or mixed injury) between groups as determined by R score 

(p=0.26). On day 1 of enrollment, conventional medicine DILI 

showed higher serum alkaline phosphatase levels compared 

to the CAM group (mean IU/L, 215.1 vs. 156.2, p=0.005); 

there were no differences between both cohorts in ALT/AST, 

INR, platelets, bilirubin or MELD score or coma grade. Despite 

fewer individual comorbidities were noted in the CAM population 

(2.03 vs. 1.15, p=0.005), the CAM group had a lower 

transplant-free survival at 21 days, though this was not statistically 

significant (24.3% vs. 38.5%, p=0.10). Within the CAM 

group, those surviving without transplantation had significantly 

lower INRs (1.9 vs. 3.8, p


213 

Safety and Tolerability of Ornithine Phenylacetate to 

Lower Ammonia in Acute Liver Failure: Preliminary 

report of the STOP-ALF Trial 

William M. Lee 1 , R. Todd Stravitz 2 , Robert J. Fontana 3 , A. James 

Hanje 4 , Holly Tillman 5 , Kristen M. Clasen 5 , Valerie L. Durkalski 5 , 

Averell H. Sherker 6 , Edward Doo 6 , Brian C. Davis 7 ; 1 Division of 

Digestive and Liver Diseases, University of Texas Southwestern 

Medical Center at Dallas, Dallas, TX; 2 Gastroenterology, Virginia 

Commonwealth University, Richmond, VA; 3 Gastroenterology, University 

of Michigan, Ann Arbor, MI; 4 Gastroenterology, Hepatology 

and Nutrition, The Ohio State University Wexner School of 

Medicine, Columbus, OH; 5 Medical University of South Carolina, 

Charleston, SC; 6 NIDDK, NIH, Bethesda, MD; 7 Internal Medicine, 

UT Southwestern Medical Center at Dallas, Dallas, TX 

Background: No satisfactory method is currently available to 

lower ammonia (NH 3 

) to prevent or treat hepatic encephalopathy 

(HE) and cerebral edema in hospitalized patients with 

acute liver injury (ALI) or acute liver failure (ALF). Ornithine 

phenylacetate (OPA, OCR-002) effectively lowers NH 3 

in animal 

models of ALF by trapping it as phenylacetylglutamine 

(PAGN) that is excreted in urine. STOP-ALF is a phase 2a trial 

to develop data on safety and tolerability of a 5-day infusion 

of OCR-002 in the setting of ALF/ALI and elevated NH 3 

levels. 

Methods: Patients with ALF or ALI and elevated NH 3 

levels, 

>60 mmol/L, were enrolled at 8 US sites participating in the 

Acute Liver Failure Study Group (ALFSG) in two cohorts: normal/minimal 

renal dysfunction (Cr < 1.5, Cohort 1) and acute 

kidney injury (AKI, Cr ≥ 1.5 mg/dL, Cohort 2). Dosing of OCR- 

002 was open label, ascending dose, starting at 3.3 g/24hr 

by IV infusion, to a maximum dose of 10 gm/24 hr after 24 

hr of infusion; for the 1st 3 pts, final dose = 3.3 gm/24hX5 

days, then 6.7 gm/24hr for 3 and then 10 gm/24h for the 

remaining patients in each cohort. Plasma and urine PK and 

plasma NH 3 

levels were recorded, in addition to safety labs, 

EKGs and frequent neurological checks. Results: 30 patients 

have been enrolled, with current analysis based on 22 with PK 

data available; median age 32 yrs, 14/22 female, 16 were 

acetaminophen-related, 17 were considered evaluable, having 

received >72 hrs of OCR-002. The drug was well tolerated 

with only minor AE’s attributed (nausea, headache). PK results 

showed mean phenylacetate (PAA) plasma levels of 29.0 and 

62.5 mg/mL in Cohort 1 at the 2 higher dose levels, and 32.5 

mg/mL for Cohort 2, dose level 2. These levels of PAA are 

below the optimal concentration for NH 3 

removal. Overall survival 

was 73%, with deaths limited to advanced AKI patients 

and those unable to receive liver transplantation; 1 pt. required 

transplantation. NH 3 

levels improved in most patients: median 

baseline NH 3 

=87 mmol/L to 54 mmol/L day 4, 0 hour. This 

ongoing trial is currently completing the final dose level; no 

toxic levels of PAA or PAGN have been reached. Conclusion: 

OCR-002 was well tolerated without any safety signals and 

shows promise as adjunctive therapy in ALF. Higher doses of 

OCR-002 may be needed for optimal ammonia reduction in 

ALF. Future studies to assess safety and efficacy of higher doses 

of OCR-002 will be pursued. 

Disclosures: 

William M. Lee - Consulting: Eli Lilly, Sanofi; Grant/Research Support: Gilead, 

BMS, Vertex, Merck 

Robert J. Fontana - Consulting: GlaxoSmithKline, CLDF; Grant/Research Support: 

Gilead, vertex, BMS, Jansen, Gilead 

A. James Hanje - Consulting: Salix pharmaceutical 

The following authors have nothing to disclose: R. Todd Stravitz, Holly Tillman, 

Kristen M. Clasen, Valerie L. Durkalski, Averell H. Sherker, Edward Doo, Brian 

C. Davis 

214 

Characterization of Cerebral Edema in Acute on Chronic 

Liver Failure 

Radha K. Dhiman 1 , Tarana Gupta 1 , Chirag K. Ahuja 2 , Swastik 

Agrawal 1 , Naveen Kalra 2 , Ajay K. Duseja 1 , Niranjan Khandelwal 

2 , Yogesh K. Chawla 1 ; 1 Hepatology, Postgraduate Institute of 

Medical Education & Research, Chandigarh, India; 2 Radiodiagnosis 

& Imaging, Postgraduate Institute of Medical Education & 

Research, Chandigarh, India 

Background and Aims: The significance and nature of cerebral 

edema in acute-on-chronic liver failure (ACLF) is not well 

studied. This study aimed to characterize cerebral edema in 

ACLF using magnetization transfer ratio (MTR) and diffusion 

tensor imaging (DTI). Methods: Forty consecutive patients with 

cirrhosis and acute decompensation were included. Patients 

were categorized as no-ACLF (n=11), grade 1 (n=10), grade 

2 (n=9) and grade 3 (n=10) ACLF as per CANONIC study criteria 

(Moreau et al. Gastroenterology 2013;144:1426-1437). 

Whole brain conventional magnetic resonance imaging (MRI) 

was performed on a 3.0 Tesla scanner using standard 16/32 

channel head coils. Apart from the conventional sequences 

namely T2 weighted, FLAIR weighted and T1 weighted volumetric 

sequences (MPRAGE/SPGR BRAVO), additional sequences 

performed were magnetization transfer ratio MTR and DTI. 

MRI of brain was performed and tumor necrosis factor α, interleukin 

(IL)-1β, and IL-6 were measured at baseline and after 

7-10 days of admission. Patients were followed up for 90-days 

from admission. Ten age- and sex-matched healthy subjects 

were also included as controls. Results: MTR values were significantly 

lower in patients in both no-ACLF and ACLF groups 

compared to controls. Mean diffusivity (MD) values progressively 

increased from controls to no-ACLF to ACLF grade 1, 2 

and 3 groups in the basal ganglia (P 8 x 10 -9 M 2 /s) in frontal white matter were independent 

predictors of 90-day mortality (Table). Conclusions: Our 

study conclusively demonstrates cerebral edema increases with 

severity of ACLF and is predominantly due to extracellular brain 

water increase. Correlation between MD values and IL-6 levels 

suggests pathogenic role of inflammation. MELD score and 

cerebral edema have prognostic significance in ACLF patients. 

Utility of MELD and mean diffusivity values as predictors of 

90-day mortality 

MELD, model for end-stage liver disease; MD FWM, mean diffusivity 

frontal white matter; PPV, positive predictive value; NPV, 

negative predictive value; DA, diagnostic value, AUROC, area 

under ROC curve 

Disclosures: 

The following authors have nothing to disclose: Radha K. Dhiman, Tarana Gupta, 

Chirag K. Ahuja, Swastik Agrawal, Naveen Kalra, Ajay K. Duseja, Niranjan 

Khandelwal, Yogesh K. Chawla


215 

The transportable machine perfusion Airdrive®, a novel 

approach to safely expand the donor pool for liver 

transplantation 

Philippe Compagnon 1,2 , Ismail Ben Mosbah 2 , Hassen Hentati 1 , 

Eric Levesque 3 , Mara Disabato 1 , Jose L. Cohen 2 , Daniel Azoulay 1 ; 

1 Service de Chirurgie Digestive et Hépatobiliaire -Transplantation 

Hépatique, Hopital Henri Mondor - Assistance Publique-Hopitaux 

de Paris, Université Paris-Est, Creteil, France; 2 IMRB- Inserm U955, 

Hopital Henri Mondor, Assistance Publique-Hopitaux de Paris, Université 

Paris-Est créteil, Créteil, France; 3 Service de Réanimation 

Digestive, Hopital Henri Mondor, Assitance Publique-Hopitaux de 

Paris, Université Paris-Est, Créteil, France 

Background and Aims: Livers derived from donation after 

circulatory death (DCD) suffer warm ischemia (WI) and are 

infrequently used for transplantation; they have the potential, 

however, to considerably expand the donor pool. The 

Airdrive® is the first transportable oxygenated machine perfusion 

(MP) unit suitable for liver preservation. We aimed 

to determine whether the Airdrive® MP would improve the 

quality of these livers using a DCD large animal model. Methods: 

Female large white pigs were used as organ donors and 

recipients. Liver allografts procured from heart beating donors 

and preserved by simple cold storage (SCS) served as controls 

(n=6). In experimental groups, cardiac arrest was induced by 

IV injection of KCL. After 60min WI, livers were perfused in 

situ with HTK and subsequently preserved either by SCS (SCS 

group, n=3) or hypothermic MP (Airdrive® group, n=4) using 

MPS-Belzer solution. After 4hr of preservation, all livers were 

transplanted into recipient pigs. The main judgment criterion 

was animal survival at day 5 Results: All animals that received 

a simple cold stored liver allograft after 60min WI experienced 

PNF and died within 6 hours after transplantation (5-day survival 

= 0%). In contrast, 5-day survival in animals that received 

a MP liver was 100% (4/4, Airdrive® group) as well as in 

controls (6/6). A post-reperfusion syndrome was observed in 

all animals (3/3) of the SCS group but none of the control 

or Airdrive® groups. These phenomena caused a significant 

increase in fluid challenge and catecholamine needs in SCS 

group relative to control and Airdrive® groups. At the end of 

cold preservation, ATP content was significantly higher in the 

Airdrive® group vs. SCS group. After reperfusion, MP livers 

functioned better (albumin production, prothrombin time rates) 

and showed less hepatocellular and endothelial cell injury, in 

agreement with better preserved liver integrity (histology) vs. 

SCS group. MP livers also exhibited higher ATP recovery than 

SCS livers. The protective effect of the Airdrive® device was 

associated with a significant attenuation of oxidative stress 

(lower lipid peroxidation, higher catalase and superoxide 

dismutase activity), and a better endoplasmic reticulum adaptation 

leading to limit mitochondrial damage (cytochrome c, 

caspase 9, GLDH), and apoptosis (caspase 3 and TUNEL) Conclusions: 

This study demonstrates for the first time the efficacy of 

the transportable MP Airdrive® device to enhance donor liver 

viability for transplantation in a clinically relevant DCD model. 

Disclosures: 

The following authors have nothing to disclose: Philippe Compagnon, Ismail Ben 

Mosbah, Hassen Hentati, Eric Levesque, Mara Disabato, Jose L. Cohen, Daniel 

Azoulay 

216 

A Novel Percutaneous Organ Perfusion Stent Improves 

Liver Perfusion in a Porcine Model of Donation after 

Cardiac Death 

Bryan Tillman 1 , Youngjae Chun 2 , Nathan L. Liang 1 , Tara D. Richards 

1 , Anthony J. Demetris 4 , Timothy M. Maul 2 , Amit D. Tevar 3 ; 

1 Division of Vascular Surgery, University of Pittsburgh Medical 

Center, Pittsburgh, PA; 2 Swanson School of Engineering, University 

of Pittsburgh, Pittsburgh, PA; 3 Starzl Transplant Institute, University 

of Pittsburgh Medical Center, Pittsburgh, PA; 4 Department of 

Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA 

Introduction: Donation after cardiac death (DCD) liver allografts 

are frequently discarded due to ischemia during the agonal 

period prior to cardiac death. A dual-chambered Organ Perfusion 

Stent (OPS) would isolate blood flow to the abdominal 

organs to maintain perfusion, while agonal cardiac flow would 

pass through a large central lumen. As a result, this approach 

would isolate perfusion to the liver from the unstable circulation 

of the agonal donor without affecting donor cardiac 

death. Methods: OPS were constructed with a central lumen 

for cardiac flow and an external visceral perfusion chamber. A 

porcine model included three control and six stent animals. The 

OPS was deployed through endovascular sheaths in the aorta 

to include the visceral branches while under cardiac monitoring. 

Venous blood from a separate cannula was cycled through 

an oxygenator and back to the isolated visceral arteries. Next, 

the DCD agonal period was simulated for 60 minutes with a 

target MAP of 40 mm Hg. Results: Angiography of the outer 

chamber confirmed perfusion of the visceral organs, while 

agonal cardiac blood flow passed through the central lumen. 

During stent placement, no change was observed in cardiac 

parameters of HR, RVEDV, CVP, MAP, or SvO2 (P > 0.05) 

(n=6). Cardiac output decreased by an average 11% consistent 

with offloading of visceral perfusion (P=0.01). Stented 

animals revealed an average of 4.8 fold improved oxygen 

delivery compared to controls. Following the agonal period, 

the livers of the controls revealed marked ischemic appearance 

contrasting to the adequately perfused livers of the stent treated 

animals. Conclusions: This novel OPS achieves improved oxygen 

delivery to visceral organs without significantly impacting 

cardiac physiology of the donor. A single deployment step 

allows for collective isolation of all four visceral branches. In 

summary, this novel approach may significantly increase the 

quality of livers available for transplantation. 

Disclosures: 

Timothy M. Maul - Advisory Committees or Review Panels: Mallinckrodt 

The following authors have nothing to disclose: Bryan Tillman, Youngjae Chun, 

Nathan L. Liang, Tara D. Richards, Anthony J. Demetris, Amit D. Tevar


217 

Daclatasvir and Sofosbuvir in Patients with Recurrent 

HCV Following Liver Transplantation and Advanced 

Fibrosis or Cirrhosis: United States Multicenter Treatment 

Protocol 

Paul Y. Kwo 1 , Michael W. Fried 2 , K. Rajender Reddy 3 , Consuelo 

Soldevila-Pico 4 , Saro Khemichian 5 , Jama M. Darling 2 , Andrew 

A. Napoli 6 , Beatrice Anduze-Faris 6 , Robert S. Brown 7 ; 1 Indiana 

University, Indianapolis, IN; 2 University of North Carolina, Chapel 

Hill, NC; 3 Department of Medicine, University of Pennsyslvania, 

Philadelphia, PA; 4 Department of Medicine, University of Florida, 

Gainesville, GA; 5 Keck School of Medicine, University of Southern 

California, Los Angeles, CA; 6 Bristol-Meyers Squibb, Plainsboro, 

NJ; 7 Department of Medicine, Columbia University College of Physicians 

& Surgeons, New York, NY 

Background: In the United States, there are limited treatment 

options for liver transplant recipients with recurrent HCV and 

advanced fibrosis/cirrhosis. In a Phase 3 study, the pangenotypic 

combination of daclatasvir (DCV) and sofosbuvir (SOF) 

with ribavirin demonstrated SVR rates of 94% in liver transplant 

recipients with HCV recurrence. This analysis reports interim 

findings from a U.S. Treatment Use protocol in patients with 

recurrent HCV following liver transplantation and advanced 

fibrosis or cirrhosis. Methods: In this multicenter, open-label, 

expanded access study, DCV was provided for use in combination 

with SOF, with or without RBV at the investigator’s 

discretion, for up to 24 weeks in genotype 1-6 patients with 

F3 fibrosis, cirrhosis, or fibrosing cholestatic hepatitis (FCH) 

following liver transplantation. The HCV TARGET consortium of 

academic and community medical centers and data collection 

methodology were utilized. This interim analysis includes 51 

patients who have enrolled and initiated treatment at 20 sites. 

Results: Of the 51 patients treated, 49 received DCV+SOF 

and 2 received DCV+SOF+RBV. Patients were predominantly 

male (77%), white (82%) and infected with HCV genotype 1 

(73%) or genotype 3 (24%). Fifty-seven percent had failed 

prior treatment with IFN-based regimens (12% included a 

protease inhibitor). Cirrhosis was present in 82% of patients, 

including 57% with evidence of post transplant hepatic decompensation, 

8% had a kidney transplant, and 8% had FCH. At 

baseline, median HCV RNA was 2.9 x 10 6 IU/ml, the mean 

(range) for platelets was 129 (41-239) x10 3 cells/uL, albumin 

3.4 (1.7-6.3) g/dL, total bilirubin 1.5 (0.3-13) mg/dL, INR 1.1 

(1.0-1.5), and creatinine clearance was 83 (40-190) ml/min. 

MELD was ≥10 in 48% of patients with available scores. Of 

21 patients with available data, the end of treatment virologic 

response was 100%. All four patients who reached post-treatment 

week 12 achieved SVR. Adverse events occurring in 

≥10% of patients were fatigue, headache, nausea, and arthralgia. 

Serious adverse events of interest included transplant rejection 

(n=1) and renal failure (n=4). Two patients discontinued 

treatment for adverse events; 1 due to renal failure leading to 

death (unrelated to treatment) and 1 due to death of unknown 

cause. SVR results for 31 patients will be available for presentation. 

Conclusion: DCV+SOF for 24 weeks was well-tolerated 

in this population with severe recurrent post-transplant hepatitis 

C. Interim data indicate effective end of treatment antiviral 

response and SVR in the patients receiving this regimen 

through compassionate use. 

Disclosures: 

Paul Y. Kwo - Advisory Committees or Review Panels: Abbvie, Abbott, Novartis, 

Merck, Gilead, BMS, Janssen; Consulting: BMS; Grant/Research Support: 

Roche, Abbvie, Merck, BMS, Abbott, Idenix, Vital Therapeutics, Gilead, Vertex, 

Merck, Idenix; Speaking and Teaching: Merck, Merck 

Michael W. Fried - Consulting: Merck, Abbvie, Janssen, Bristol Myers Squibb, 

Gilead; Grant/Research Support: Merck, AbbVie, Janssen, Bristol Myers Squibb, 

Gilead; Patent Held/Filed: HCCPlex 

K. Rajender Reddy - Advisory Committees or Review Panels: Merck, Janssen, 

Vertex, Gilead, BMS, Abbvie; Grant/Research Support: Merck, BMS, Ikaria, 

Gilead, Janssen, AbbVie 

Saro Khemichian - Advisory Committees or Review Panels: Gilead, AbbVie, BMS; 

Speaking and Teaching: Gilead, AbbVie, Salix 

Jama M. Darling - Consulting: BristolMyers Squibb; Grant/Research Support: 

BristolMyers Squibb 

Andrew A. Napoli - Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers 

Squibb 

Beatrice Anduze-Faris - Employment: Bristol-Myers Squibb 

Robert S. Brown - Consulting: Gilead, Janssen, Abbvie, Merck, BMS 

The following authors have nothing to disclose: Consuelo Soldevila-Pico 

218 

Rapid Decline In Interferon Stimulated Genes In Prior 

HCV Non-Responders Re-Treated With Direct Acting 

Antivirals 

Hawwa Alao 1 , Yun Ju Kim 1 , Elizabeth C. Wright 4 , Margaret 

Cam 2 , Elenita M. Rivera 1 , Nancy Fryzek 1 , David E. Kleiner 3 , Fang 

Zhang 1 , Edward Doo 1 , Yaron Rotman 1 , Christopher Koh 1 , Theo 

Heller 1 , Jay H. Hoofnagle 1 , T. Jake Liang 1 , Marc G. Ghany 1 ; 

1 Liver Diseases Branch, National Institutes of Health, Bethesda, 

MD; 2 NCI, NIH, Bethesda, MD; 3 LP, NCI, NIH, Bethesda, MD; 

4 Office of the Director, NIDDK, NIH, Bethesda, MD 

BACKGROUND: The development of direct acting antiviral 

(DAA) agents has revolutionized therapy of chronic hepatitis 

C. Prior non-responders are known to have high baseline 

hepatic expression of interferon stimulated genes (ISGs). The 

effect of potent DAAs on ISG expression is unknown. AIM: To 

investigate the effect of an oral DAA regimen on ISG expression 

in prior failures to interferon-based regimens with chronic 

hepatitis C (CHC). METHODS: Subjects with HCV genotype 

1b infection who had failed to respond to peginterferon and 

ribavirin were re-treated with asunaprevir anddaclatasvir for 

24 weeks. Subjects were monitored after therapy for 12 weeks 

to determine SVR12. Patients underwent paired liver biopsies, 

the first within 12 weeks before therapy and the second at 

either week 2 or 4 on therapy. Microarray analysis (Affymetrix 

Human Gene 2.0 ST array, Santa Clara, CA) was performed 

on liver biopsy tissue and on-therapy gene expression was 

compared to baseline. All subjects provided informed consent 

to participate. RESULTS: 11 patients with chronic HCV genotype 

1b infection had paired biopsies available for microarray 

analysis. The 11 patients included 6 women, mean age 

62 years, mean HCV RNA 6.8 log 10 

IU/mL, 4 with cirrhosis. 

The SVR rate was 64%. All 4 failures experienced virological 

breakthrough between weeks 4-12. At the week 2 biopsy, 5 of 

5 subjects had detectable virus (


Disclosures: 

The following authors have nothing to disclose: Hawwa Alao, Yun Ju Kim, Elizabeth 

C. Wright, Margaret Cam, Elenita M. Rivera, Nancy Fryzek, David E. 

Kleiner, Fang Zhang, Edward Doo, Yaron Rotman, Christopher Koh, Theo Heller, 

Jay H. Hoofnagle, T. Jake Liang, Marc G. Ghany 

219 

The Emergence of NS5B Resistant Associated Variant 

S282T after Sofosbuvir-Based Treatment 

Edward J. Gane 2 , Armando Abergel 3 , Sophie Metivier 4 , Ronald 

Nahass 5 , Michael Ryan 6 , Catherine A. Stedman 7 , Evguenia S. 

Svarovskaia 1 , Hongmei Mo 1 , Brian Doehle 1 , Hadas Dvory-Sobol 1 , 

Charlotte Hedskog 1 , Ming Lin 1 , Diana M. Brainard 1 , Phillip S. 

Pang 1 , Jenny C. Yang 1 , John G. McHutchison 1 , Mark S. Sulkowski 

8 , Ziad Younes 9 , Eric Lawitz 10 ; 1 Gilead Sciences, Foster City, 

CA; 2 Auckland Clinical Studies, Auckland, New Zealand; 3 Centre 

Hospitalier Universitaire-Estaing, Clermont-Ferrand, France; 4 Centre 

Hospitalier Universitaire-Purpan, Toulouse, France; 5 ID Care, 

Inc, Hillsborough, NJ; 6 Digestive and Liver Disease Specialists, 

Norfolk, VA; 7 Christchurch Clinical Studies Trust, Christchurch, 

New Zealand; 8 Johns Hopkins University School of Medicine, Baltimore, 

MD; 9 Gastro One, Germantown, TN; 10 University of Texas 

Health Science Center, Texas Liver Institute, San Antonio, TX 

Introduction: Sofosbuvir (SOF), a nucleotide prodrug, and ledipasvir/sofosbuvir 

(LDV/SOF) are approved for the treatment of 

chronic HCV. In vitro HCV replicon results demonstrated that 

the NS5B substitution, S282T, confers a 2- to 18-fold reduced 

susceptibility to SOF in all genotypes while also reducing the 

viral replication capacity by 89% to 99% when compared to 

wild-type. The aim of this analysis is to evaluate the emergence 

and fitness of the S282T variant in virologic failure patients 

administered SOF-based regimens across the SOF and LDV/ 

SOF Phase 2 and Phase 3 programs. Methods: Plasma samples 

collected prior to SOF treatment and at the time of virologic 

failure were evaluated. The HCV NS5B region was amplified 

and deep sequenced (cut-off at 1%). Whole HCV genome 

(core to NS5B) sequencing and replication capacity assays 

were performed for a subset of patients with S282T to investigate 

presence of compensatory substitutions. Results: To date, 

over 12,000 patients have been treated in SOF or LDV/SOF 

Phase 2 and 3 studies. Of these, deep sequencing was available 

at baseline in 6,086 subjects (70% GT1, 8% GT2, 18% 

GT3, and 4% GT4-6) and at virologic failure in 901 subjects. 

SOF regimens administered included monotherapy, SOF+RB- 

V±Peg-IFN, or LDV/SOF±RBV for up to 24 weeks. At baseline, 

no (0/6086) S282T variant was detected. At virologic failure, 

10 of 901 (1%) subjects had S282T detected and 6 of these 

subjects had additional NS5B treatment-emergent variants 

(L159F, E237G, M289L/I, L320V/I, and V321I) detected. In 

the 4 subjects with S282T for whom the entire HCV genome 

was successfully sequenced, no consistent amino acid substitutions 

were observed outside NS5B. Replication capacity testing 

in vitro also did not identify any restoration of the replicative 

defect of S282T. Subjects with S282T received SOF monotherapy 

(n=2), retreatment with LDV/SOF in prior LDV/SOF 

failures (n=3), and LDV/SOF for 8 weeks in genotype 1 (n=1) 

and 12 weeks in genotypes 3, 4, 5, and 6 (n=4). S282T levels 

declined on average by 4-fold within 2 weeks of follow-up 

period confirming low replication fitness of this variant. To 

date, 4 of these patients have been retreated: with SOF+RBV in 

GT2 infected patient (n=1) or with LDV/SOF+RBV in GT1 (n=2) 

and GT3 (n=1) infected patients; all achieved SVR12. Conclusions: 

The emergence of S282T variant was rare in patients 

who fail SOF-based regimens. S282T variants are unfit in vivo 

and rapidly decline over time. Successful retreatment of prior 

SOF-failure patients is possible in the presence of S282T variants 

with SOF+RBV±LDV regimens. 

Disclosures: 





Sophie Metivier - Speaking and Teaching: Roche, BMS, Janssen, Merck, Gilead, 

abbvie 

Ronald Nahass - Advisory Committees or Review Panels: Gilead, MErck, Janssen, 

BMS; Grant/Research Support: Gilead, Merck, Janssen, BMS; Speaking and 

Teaching: Gilead, Merck, Janssen 

Catherine A. Stedman - Advisory Committees or Review Panels: MSD, Abbvie; 

Speaking and Teaching: Gilead, Abbvie 

Evguenia S. Svarovskaia - Employment: Gilead Sciences Inc.; Stock Shareholder: 

Gilead Sciences Inc. 




Sciences 

Charlotte Hedskog - Consulting: Gilead Sciences 

Ming Lin - Employment: Gilead Sciences, Inc. 


Sciences 


Jenny C. Yang - Employment: Gilead Sciences, Inc 


Sciences 



BMS 

Ziad Younes - Consulting: Gilead; Grant/Research Support: BMS, AbbVie, Gilead, 

Vertex, Vertex, Idenix, Merck, Janssen, Tobira; Speaking and Teaching: 

Gilead, AbbVie 

Eric Lawitz - Advisory Committees or Review Panels: AbbVie, Achillion Pharmaceuticals, 

Regulus, Theravance, Enanta, Idenix Pharmaceuticals, Janssen, Merck 

& Co, Novartis, Gilead; Grant/Research Support: AbbVie, Achillion Pharmaceuticals, 

Boehringer Ingelheim, Bristol-Myers Squibb, Gilead Sciences, GlaxoSmith- 

Kline, Idenix Pharmaceuticals, Intercept Pharmaceuticals, Janssen, Merck & Co, 

Novartis, Nitto Denko, Theravance, Salix, Enanta; Speaking and Teaching: Gilead, 

Janssen, AbbVie, Bristol Meyers Squibb 

The following authors have nothing to disclose: Michael Ryan 

220 

Evolution Of Resistance Associated Variants During 

Initial Treatment, Viral Failure, And Re-Treatment With 

Directly Acting Antiviral Therapy (NIH SYNERGY Trial) 

Sarah Kattakuzhy 2,3 , Eleanor Wilson 2,3 , Sreetha Sidharthan 3 , 

Mary McLaughlin 4 , Rachel Silk 2 , Hongmei Mo 5 , Anu Osinusi 5 , 

Anita Kohli 6,1 , Henry Masur 3 , Shyam Kottilil 2,3 ; 1 Laboratory 

of Immunoregulation, NIH, Leidos Biomedical Research Inc., 

Bethesda, MD; 2 Infectious Disease, University of Maryland School 

of Medicine, Institute of Human Virology, Baltimore, MD; 3 Critical 

Care Medicine, National Institutes of Health, Bethesda, MD; 

4 National Institute of Allergy and Infectious Disease, Bethesda, 

MD; 5 Gilead Sciences, Inc., Foster City, CA; 6 Hepatology, Creighton 

University, Phoenix, AZ 

Introduction: The natural history of resistance associated 

variants (RAVs), and their impact on treatment in the era of 

directly-acting antiviral therapy (DAA) remains unclear. Recent 

studies have suggested that NS5A RAVs exhibit long-term persistence 

and may affect response to NS5A inhibitor based 

therapy. The current analysis seeks to outline the evolution 

of RAVs to the major HCV therapeutic target regions (NS3, 

NS5A, and NS5B), evaluate the viral evolution of these mutations 

over time, and assess the impact on initial treatment and 

retreatment outcomes Methods: In this single-center, open-la-


bel, phase 2a trial, 34 HCV mono-infected participants with 

F0-F2 liver fibrosis, with previous viral relapse to 4-6 weeks 

of LDV/SOF + GS-9451 +/- GS-9669, were enrolled and 

re-treated with LDV/SOF for 12 weeks. Deep sequencing of 

NS3, NS5A, and NS5B regions was performed by Illumina 

next generation sequencing technology with a detection limit of 

1% prevalence. Samples were sequenced at three time pointsprior 

to initial therapy, at the time of relapse, and prior to 

retreatment; two-way ANOVA testing was used to evaluate 

significant change in RAVs Results: 33 patient samples were 

sequenced at each timepoint. At baseline, 10 patients (30%), 

9 patients (27%) and 1 patient (3%) had RAVs to NS3, NS5A, 

and NS5B respectively. At a median of 4 weeks post treatment, 

patients experienced viral relapse to short duration therapy. 

At viral relapse, 11 patients (33%) and 2 patients (6.1%) had 

NS3 and NS5B RAVs respectively. The number of patients with 

NS5A RAVs increased significantly to 28 patients (84%), and 

emergent mutations L31M (+30.3%; 95% CI: 7.8-52.7) and 

Q30R (+27.3%; 95% CI: 4.9-49.7) demonstrated a significant 

increase from baseline to viral relapse. After a median interval 

of 22 weeks (IQR 18-23), patients were retreated. At retreatment 

baseline,10 patients (30%), 28 patients (84%), and 3 

patients (9%) had NS3, NS5A, and NS5B RAVs respectively, 

with no significant change in profile of mutants compared to 

the time of viral relapse. There was no significant difference in 

number of RAVs per sample along the three timepoints. In this 

retreatment cohort, 31 patients out of 34 (91% ITT; 96%mITT) 

achieved sustained virologic response Conclusions: In this small 

cohort, there was a moderate baseline rate of NS3 and NS5A 

resistance, and minimal NS5B resistance. While failure to short 

duration therapy was characterized by persistent and emergent 

NS5A mutants, retreatment outcomes with longer duration therapy 

were not affected by the presence of these RAVs 

Disclosures: 



The following authors have nothing to disclose: Sarah Kattakuzhy, Eleanor 

Wilson, Sreetha Sidharthan, Mary McLaughlin, Rachel Silk, Anita Kohli, Henry 

Masur, Shyam Kottilil 

221 

Polymorphisms in irisin-producing FNDC5 are associated 

with the severity of HCV genotype 3-induced steatosis 

Stephanie Bibert 2 , Claudio De Vito 3 , Beat Mullhaupt 4 , David 

Semela 5 , Jean-Francois Dufour 6 , Markus H. Heim 7 , Darius 

Moradpour 8 , Andreas Cerny 9 , Raffaele Malinverni 10 , Pierre-Yves 

Bochud 2 , Francesco Negro 1 ; 1 Gastroenterology, Hepatology and 

Clinical pathology, University Hospitals, Geneva-14, Switzerland; 

2 Service of Infectious Diseases, University Hospital, Lausanne, Switzerland; 

3 Clinical pathology, University Hospital, Geneva, Switzerland; 

4 Gastroenterology and hepatology, University Hospital, 

Zurich, Switzerland; 5 Gastroenterology, Cantonal Hospital, St. 

Gallen, Switzerland; 6 Visceral surgery and medicine, Inselspital, 

Berne, Switzerland; 7 Gastroenterology, Univresity Hospital, Basel, 

Switzerland; 8 Gastroenterology, University Hospital, Lausanne, 

Switzerland; 9 Epatocentro, Lugano, Switzerland; 10 Medicine, 

Pourtalès Hospital, Neuchatel, Switzerland 

Background. Steatosis, commonly observed among chronically 

HCV-infected patients, is associated with an increased risk of 

hepatocellular carcinoma development and cardiovascular morbidity. 

While metabolic steatosis is associated with changes in 

host metabolism including obesity, hyperlipidemia and insulin 

resistance, steatosis in HCV genotype 3 infection is triggered 

by the virus. Hence, both host and viral factors contribute to the 

development of steatosis. To date, only few host genetic polymorphisms 

have been associated with HCV-related steatosis. 

Single nucleotide polymorphisms (SNPs) within the gene coding 

for fibronectin type II domain-containing 5 transmembrane 

receptor (FNDC5) have recently been linked to NAFLD in Caucasian 

patients. Moreover, recent studies showed that irisin, the 

cleaved and secreted form of FNDC5, could prevent hepatic 

steatosis in NAFLD patients. Methods. We analyzed the role 

of 4 FNDC5 gene-tagging polymorphisms as well as PNPLA3 

rs738409, IFNL3 rs12980275 and TM6SF2 rs3794991 in 

548 Caucasian HCV-infected patients with available liver 

biopsy. SNPs were extracted from a genome-wide association 

dataset. Genetic associations with the presence and severity 

of steatosis were assessed by univariate and multivariate logistic 

regression after adjustment for relevant covariates. Results. 

We confirm that PNPLA3 rs738409, IFNL3 rs12980275, and 

TM6SF2 rs3794991 are independent predictors of steatosis 

in 548 patients with HCV genotypes other than 3. In addition, 

we found that carriage of an SNP in FNDC5 increases the 

risk of steatosis only in patients infected with HCV genotype 

3 (odds ratio [OR] = 7.08, 95% confidence interval [CI] = 

1.44-34.82, p = 0.016) and appears to be associated with 

the severity of steatosis (OR = 5.45, 95% CI = 1.46-20.40, p 

= 0.012), although the limited number of patients prevented us 

from drawing firm conclusions. Conclusion. Our data suggest 

that FNDC5 could be specifically involved in the development 

and severity of steatosis in HCV genotype 3 infection. 

Disclosures: 

Beat Mullhaupt - Consulting: MSD, Novartis, MSD, Janssen; Grant/Research 

Support: Bayer, Gillead 

Jean-Francois Dufour - Advisory Committees or Review Panels: Bayer, BMS, Gilead, 

AbbVie, Novartis, Sillagen, Genfit 

Pierre-Yves Bochud - Speaking and Teaching: MSD, Gilead 

Francesco Negro - Advisory Committees or Review Panels: Bristol-Myers Squibb, 

MSD, Gilead, AbbVie; Grant/Research Support: Gilead 

The following authors have nothing to disclose: Stephanie Bibert, Claudio De 

Vito, David Semela, Markus H. Heim, Darius Moradpour, Andreas Cerny, Raffaele 

Malinverni 

222 

Fast and deep early HCV-RNA decay is accompanied by 

fast aminotransferases normalization values in cirrhotic 

HCV-1 patients treated with paritaprevir/r–ombitasvir 

and dasabuvir with ribavirin: a potential role of baseline 

major resistance mutations 

Valeria Cento 1 , Elisabetta Teti 2 , Elisa Biliotti 3 , Francesco Fiore 7 , 

FrancescoPaolo Antonucci 1 , Marianna Aragri 1 , Carlo F. Magni 4 , 

Daniele Di Paolo 5 , Donatella Palazzo 3 , Katia Yu La Rosa 1 , Francesca 

De Luca 1 , Maria Chiara Sorbo 1 , Velia Chiara Di Maio 1 , 

Paolo Casalino 6 , Cesare Sarrecchia 2 , Lara Lambiase 7 , Laura 

Gianserra 7 , Paola Perinelli 3 , Loredana Sarmati 2 , Guido A. Gubertini 

4 , Mario Angelico 5 , Caterina Pasquazzi 7 , Massimo Andreoni 2 , 

Gloria Taliani 3 , Carlo Federico Perno 1 , Francesca Ceccherini-Silberstein 

1 ; 1 Experimental Medicine and Surgery, University of Rome 

“Tor Vergata”, Rome, Italy; 2 Infectious Diseases, Polyclinic “Tor 

Vergata” Foundation, Rome, Italy; 3 Infectious and Tropical diseases, 

“Umberto I” Hospital, Rome, Italy; 4 Division of Infectious 

Disease, Hospital Sacco of Milan, Milan, Italy; 5 Hepatology, Polyclinic 

“Tor Vergata” Foundation, Rome, Italy; 6 Laboratory Medicine, 

Polyclinic “Tor Vergata” Foundation, Rome, Italy; 7 Infectious 

Diseases, Sant’Andrea Hospital, Rome, Italy 

Study purpose. This study aims to define in a real-life setting 

the early kinetics of HCV-RNA decay and hepatic “cell cure” 

during administration of paritaprevir/r-ombitasvir-dasabuvir 

(3D regimen) with ribavirin in cirrhotic patients (pts), and 

to investigate the role of pre-existing resistance associated


variants (RAVs). Methods. 22 HCV-1 (1a/1b/1g=7/14/1) 

Child A/B (N=20/2) cirrhotic pts received 3D plus ribavirn 

for 12/24 weeks. Baseline (BL) presence of NS3-NS5A-NS5B 

RAVs was assessed by direct sequencing. HCV-RNA and aminotransferases 

(AT) were quantified at BL and early time-points 

(4h-6/8h-24h-48h-72h-96h-1w-2w-3w-4w). Ad interim results 

at 4w-treatment are presented. Results. BL median (IQR) HCV- 

RNA was 5.7 (5.3-6.2) logIU/ml and 20 pts had high AT. 

Already at 8h after 3D-administration, a fast HCV-RNA decay 

>1 log IU/ml was observed in 13/15 (87%) pts; at that time, 

12% of pts had AT values already normalized, while another 

57% showed a remarkable drop in their value compared to 

BL. By 24h, the median (IQR) HCV-RNA decay was 2.8 (2.4- 

3.0) logIU/ml; AT normalization occurred in 75% of pts and 

an additional 11% showed a remarkable reduction in their 

values. By 1w, HCV-RNA decay slowed down (median [IQR] 

BL-1w decay= 3.5 [3.0-3.8] logIU/ml). Of great relevance, 

all pts at this time had AT within normal range. At BL, single 

3D-RAVs were detected in 6/18 pts ( NS5A 

Q30R/L31M/ 

A92T; NS5B 

C316N; NS3 

V36L/R155K), and double 3D-RAVs in 

3 (2pts: NS5B 

L159F+C316N; 1pt: NS5B 

C316N+ NS5A 

P58S). By 

8h, the 2 pts with major RAVs ( NS5A 

L31M; NS3 

R155K) were the 

only with 0.6log HCV-RNA decay vs. median (IQR) HCV-RNA 

decay 1.6 (1.4-1.9) logIU/ml in other pts (p=0.03). The same 

trend was maintained at 24h (median [IQR] HCV-RNA decay 

BL-24h=2.3 [2.3-2.4] vs. 2.8 [2.5-3.0] logIU/ml; p=0.12). At 

1w, HCV-RNA still >1000 IU/ml was found only in the 2 pts 

with 2 BL major RAVs; none achieved RVR. RVR achievement 

was positively predicted by early HCV-RNA kinetics at 48h 

and 2w of 3D administration, with a trend for BL HCV-RNA 

(p 48h 

=0.010, p 2w 

=0.003, p BL 

=0.14). Conclusions. In cirrhotic 

HCV-1 pts, 3D treatment leads to a fast and deep HCV-RNA 

decay, along with a remarkably fast normalization of AT 

values, consistent with the hypothesis of a drug-driven rapid 

“cure” of infected cells (HCV has no latent reservoir), rather 

than their elimination. Major BL RAVs may affect the kinetics of 

decay, but not necessarily the virological outcome. This result 

warrants further studies, due to the potential importance that 

rare, but specific major NS3/NS5A/NS5B RAVs may have in 

term of treatment duration and success rate of all-oral regimens. 

Disclosures: 

Mario Angelico - Advisory Committees or Review Panels: Gilead, Janssen; 

Grant/Research Support: Roche; Speaking and Teaching: GSK, Roche, Gilead, 

Novartis, BMS, Bayer 

Gloria Taliani - Speaking and Teaching: ROCHE, Merck, BMS, Gilead, Jannsen, 

Novartis, AbbVie 

Francesca Ceccherini-Silberstein - Consulting: Gilead, ViiV; Grant/Research Support: 

Merck Sharp & Dohme; Speaking and Teaching: Janssen, Abbvie, Roche 

diagnosticis 

The following authors have nothing to disclose: Valeria Cento, Elisabetta Teti, 

Elisa Biliotti, Francesco Fiore, FrancescoPaolo Antonucci, Marianna Aragri, 

Carlo F. Magni, Daniele Di Paolo, Donatella Palazzo, Katia Yu La Rosa, Francesca 

De Luca, Maria Chiara Sorbo, Velia Chiara Di Maio, Paolo Casalino, 

Cesare Sarrecchia, Lara Lambiase, Laura Gianserra, Paola Perinelli, Loredana 

Sarmati, Guido A. Gubertini, Caterina Pasquazzi, Massimo Andreoni, Carlo 

Federico Perno 

223 

Rescue of hepatocytes from lethal mitochondrial damage 

by targeted transplantation of mitochondria 

Nidhi Gupta, David Wu, Catherine H. Wu, George Wu; UCONN 

Health, Farmington, CT 

Background: Mitochondrial defects in hepatocytes can result 

in severe liver dysfunction and death. There is no method to 

repair or replace damaged mitochondria. Hepatocytes have 

cell surface asialoglycoprotein receptors (AsGR) which internalize 

asialoglycoproteins within endosomes. Some bacteria produce 

endosomolytic peptides to escape endosomes. The aim of 

this study was to determine whether hepatocytes with destroyed 

mitochondria could be rescued from cell death by targeting 

functional mitochondria by AsGR-mediated endocytosis. Methods: 

An asialoglycoprotein, asialo-orosomucoid (AsOR), was 

fluorescently labeled and covalently linked to polylysine to create 

a conjugate Fl-AsOR-PL that could bind mitochondria to form 

complexes. Huh 7 AsGR (+) and SK Hep1 AsGR (-) cells were 

treated with a mitochondrial toxin to form Huh 7-mito (-) and 

SK Hep1-mito (-) cells which required supplemental medium for 

survival. An endosomolytic peptide, LLO, was conjugated to 

AsOR to form AsOR-LLO. Complexed mitochondria and AsOR- 

LLO were co-incubated with cells for 2 h without supplemental 

media. Fluorescence, mitochondrial and cell DNA, and mitochondrial 

respiration were measured at various times. Results: 

Fl-AsOR-PL conjugate bound stably to mitochondria. Incubation 

of Fl-AsOR-PL-mitochondria complexes with Huh 7-mito (-), but 

not SK Hep1-mito (-) cells increased fluorescence in Huh7-mito 

(-) cells from 6,400 at 1 h to 11,000 units at 2 h. Co-administration 

of complexed mitochondria with AsOR-LLO conjugate 

in Huh 7-mito (-) cells increased fluorescence from 14,000 at 

1 h to >30,000 units at 2 h (p 9,700-fold over control at 7 d 

(p90% of 

controls by 10 d. Conclusions: This is the first demonstration of 

normalization of mitochondrial respiration, and rescue of mitochondria-damaged 

hepatocytes by targeted uptake of normal 

mitochondria by receptor-mediated endocytosis. 

Disclosures: 

The following authors have nothing to disclose: Nidhi Gupta, David Wu, Catherine 

H. Wu, George Wu 

224 

Predicting In Vivo Hepatotoxicity for a Pair of Related 

Compounds with a Mechanistic Model of Drug-induced 

Livery Injury 

Diane M. Longo 1,2 , Yuching Yang 1,2 , Brett A. Howell 1,2 , Scott Q. 

Siler 1,2 , Paul B. Watkins 1,2 ; 1 The Hamner-UNC Institute for Drug 

Safety Sciences, Research Triangle Park, NC; 2 DILIsym® Services, 

Research Triangle Park, NC 

Background: Tolcapone and entacapone are catechol-O-methyltransferase 

(COMT) inhibitors developed for the treatment 

of Parkinson’s disease. While clinical studies have revealed 

the potential of tolcapone to cause hepatotoxicity, no toxicity 

issues have been reported with entacapone. In vitro assays 

have shown that both compounds are capable of uncoupling 

the mitochondria proton gradient in a dose-dependent manner. 

In addition, both drugs cause modest inhibition of the bile salt 

export pump (BSEP). Aim: To integrate pharmacokinetic data 

and in vitro toxicity data via the use of a multi-scale, mechanistic 

model to simulate the in vivo response in humans to tolcapone 

and to entacapone. Methods: DILIsym ® , a predictive, mechanistic, 

mathematical model of drug-induced liver injury (DILI), was


used to simulate liver drug concentrations and DILI responses 

after administration of tolcapone (200mg, 3 times per day) 

or entacapone (200mg, 8 times per day) in a virtual human 

population sample (SimPops) of 229 simulated individuals. 

The DILIsym ® model consists of various smaller submodels that 

are mathematically integrated to simulate an organism-level 

response. The current work utilized several components of 

DILIsym®, including a physiologically-based pharmacokinetic 

(PBPK) submodel representing drug distribution, submodels of 

mitochondrial dysfunction and toxicity, bile acid physiology 

and pathophysiology, hepatocyte life cycle, and liver injury 

biomarker submodels. The DILIsym ® model is developed and 

maintained through the DILI-sim Initiative, which is a public-private 

partnership involving scientists in academia, industry, and 

the FDA. Results: In the simulated population of humans, tolcapone 

administration resulted in hepatocellular ATP reductions 

and increases in serum alanine transaminase >3x the upper 

limit of normal in 2% of the population. In contrast, entacapone 

administration did not elicit any hepatotoxicity in the 

simulated human population. This difference in hepatotoxic 

potential between tolcapone and entacapone is consistent with 

clinical observations. Conclusions: Our results demonstrate the 

utility of applying mechanistic modeling to distinguish hepatotoxic 

drugs from safer alternatives. This study also illustrates 

the capability of DILIsym ® to combine clinical data, in vitro 

data, predicted liver compound exposure, and inter-patient 

differences to provide an account of how compound exposure, 

biological variability, and multiple hepatotoxicity mechanisms 

may come together to result in DILI. 

Disclosures: 




The following authors have nothing to disclose: Diane M. Longo, Yuching Yang, 

Brett A. Howell, Scott Q. Siler 

225 

HLA-A*33:01 is strongly associated with drug-induced 

liver injury (DILI) due to terbinafine and several other 

unrelated compounds 

Guruprasad P. Aithal 1 , Paola Nicoletti 2 , Einar Bjornsson 3 , M. I. 

Lucena 23,4 , Raul J. Andrade 23,4 , Jane Grove 1 , C. Stephens 23 , Par 

Hallberg 5 , Anke H. Maitland-van der Zee 28 , Jennifer H. Martin 6,7 , 

Ingolf Cascorbi 8 , John F. Dillon 9 , Tarja Laitinen 10 , Dominique 

G. Larrey 11 , Mariam Molokhia 12 , Gerd A. Kullak-Ublick 13 , Luisa 

Ibanez 14 , Munir Pirmohamed 15 , Shengying Qin 16 , Ashley Sawle 2 , 

Fernando Bessone 21 , Nelia Hernández 22 , Andrew Stolz 24 , Naga 

P. Chalasani 25 , Jose Serrano 26 , Huiman X. Barnhart 27 , Robert J. 

Fontana 17 , Paul Watkins 18 , Thomas J. Urban 19 , Ann K. Daly 20 ; 

1 NIHR Nottingham Digestive Diseases Biomedical Research Unit, 

Nottingham University Hospitals NHS Trust, Nottingham, United 

Kingdom; 2 Columbia University, New York, NY; 3 Landspitali University 

Hospital, Reykjavik, Iceland; 4 CIBERehd, Madrid, Spain; 

5 Uppsala University, Uppsala, Sweden; 6 University of Queensland, 

Brisbane, QLD, Australia; 7 Princess Alexandra Hospital, Brisbane, 

QLD, Australia; 8 University Hospital Schleswig-Holstein, Kiel, Germany; 

9 Ninewells Hospital and medical school, Dundee, United 

Kingdom; 10 Helsinki University Central Hospital, Helsinki, Finland; 

11 Hôpital Saint Eloi, Montpellier, France; 12 Kings College London, 

London, United Kingdom; 13 University of Zurich, Zurich, Switzerland; 

14 Hospital Universitari Vall d’Hebron, Barcelona, Spain; 

15 University of Liverpool, Liverpool, United Kingdom; 16 Shanghai 

Jiao Tong university, Shanghai, China; 17 University of Michigan, 

Ann Arbor, MI; 18 Hamner-UNC institute for Drug Safety Sciences, 

Durham, NC; 19 UNC Eshelman School of Pharmacy, Chapel 

Hill, NC; 20 Newcastle University, Newcastle, United Kingdom; 

21 Facultad de ciencias Medicas, Universidad nacional de Rosario, 

Rosario, Argentina; 22 Universidad de la Republica, Mentevideo, 

Uruguay; 23 University of Malaga, Malaga, Spain; 24 University of 

Southern California, Los Angeles, CA; 25 Indiana University, Indianapolis, 

IN; 26 National Institute of Diabetes and Digestive and 

Kidney Diseases, Bethesda, MD; 27 Duke Clinical Research Institute, 

Durham, NC; 28 Utrecht University, Utrecht, Netherlands 

Background: Recent genome wide association (GWA) studies 

have revealed a number of HLA alleles as risk factors for DILI 

from specific drugs. However, untill now, the only genomewide 

significant signal seen in large DILI cohorts containing 

cases related to a range of different drugs have been HLA 

signals due to DILI from amoxicillin-clavulanate and flucloxacillin. 

Aim: To perform a GWA analysis of DILI cases enrolled by 

iDILIC and DILIN investigators that were not related to amoxicillin-clavulanate 

or flucloxacillin. Methods: Principal component 

analysis showed that 878 Caucasian cases (mean age 53 

years, 338 males; 540 females) clustered in Italians, Spanish 

and Northern Europeans groups could be matched with 

10605 population based controls. Cases and controls were 

genotyped at different times using three platforms (Illumina 1M, 

HCE and OMX). To compare the different cohorts, the genotyped 

datasets were phased and imputed. For each individual, 

HLA alleles were also predicted. After applying quality checks, 

5,429,170 variants and 217 HLA alleles were tested for association. 

We set the genomewide significance p-value threshold 

to 5*10 -8 . Results: An asociation was demonstrated between a 

single nucleotide polymorphism (SNP) that tags HLA-A*33:01 

and DILI treated as a single phenotype with an odds ratio 

(OR) of 2.65 [95%CI 1.87-3.76] (p=3.9x10 -8 ). This association 

was consistent across Italian, Spanish and Northern Europeans 

populations. When subdivided by DILI phenotype, the 

genomewide significant signal was seen only for a cholestatic/ 

mixed DILI group and not for hepatocellular DILI. Further analysis 

showed that HLA-A*33:01 was a strong risk factor for 

DILI due to the anti-fungal agent terbinafine (n=14, OR 40.53


[95%CI 12.51-288.9] (p=6.7x10 -10 )). Associations were also 

found between the risk allele and DILI from fenofibrate (n=7, 

OR 58.7[95%CI 12.31-279.8](p=3.2x10-7)) and ticlopidine 

(n=5, OR 163.1[95%CI 16.2-1642.0] (p=0.00002)). Suggestive 

signals were found also for sertraline (n=5), enalapril 

(n=4), methyldopa (n=4) and erythromycin (n=10). We confirmed 

by sequence-based typing 99% of the predictions for 34 

of 46 samples whose DILI was due to those seven drugs. HLA 

typing of a limited set of additional cases (n=13) showed 4 

further HLA-A*33 positive cases relating to terbinafine and sertraline. 

Conclusions: The novel genome-wide significant association 

of HLA-A*33:01 with all cause DILI coupled with its 

strong association with liver injury from terbinafine and other 

unrelated compounds confirms an important role for adaptive 

immunity in DILI pathogenesis and widens the range of drugs 

associated with idiosyncratic DILI for which HLA is a risk factor. 

Disclosures: 






Mariam Molokhia - Grant/Research Support: SAEC 

Fernando Bessone - Advisory Committees or Review Panels: Schering Plough, 

Gilead, Glaxo, MSD, Janssen; Speaking and Teaching: Bristol Myers Squibb, 

Janssen, Bayer, Gilead, Abbvie 

Naga P. Chalasani - Consulting: Abbvie, Lilly, Celgene, Tobira, NuSirt, Takeda, 

Merck/Anthem, Salix; Grant/Research Support: Intercept, Gilead, Galectin 



The following authors have nothing to disclose: Guruprasad P. Aithal, Paola 

Nicoletti, Einar Bjornsson, M. I. Lucena, Raul J. Andrade, Jane Grove, C. Stephens, 

Par Hallberg, Anke H. Maitland-van der Zee, Jennifer H. Martin, Ingolf 

Cascorbi, John F. Dillon, Tarja Laitinen, Gerd A. Kullak-Ublick, Luisa Ibanez, 

Munir Pirmohamed, Shengying Qin, Ashley Sawle, Nelia Hernández, Andrew 

Stolz, Jose Serrano, Huiman X. Barnhart, Paul Watkins, Thomas J. Urban, Ann 

K. Daly 

226 

Sodium 4-phenylbutyric acid protects against mice from 

acetaminophen-induced liver injury through attenuation 

of endoplasmic reticulum stress. 

Hiromi Kusama, Kazuyoshi Kon, Kenichi Ikejima, Akira Uchiyama, 

Kumiko Arai, Shunhei Yamashina, Sumio Watanabe; Gastroenterology, 

Juntendo University School of Medicine, Tokyo, Japan 

Background: Acetaminophen (AAP) overdose consequently 

elicits massive necrosis of hepatocytes, resulting in acute liver 

failure. Recent lines of reports have suggested that endoplasmic 

reticulum (ER) stress is contributed to AAP hepatotoxicity; however, 

therapeutic targeting toward ER stress has not been well 

evaluated. Therefore, here we investigate the effect of 4-phenylbutyric 

acid (PBA), a chemical chaperone, on AAP-induced 

liver injury in mice. Methods: Male, 8 week-old C57Bl/6 mice 

were exposed to AAP (450 mg/kg BW, i.p.), and serially sacrificed 

up to 12 h later. Some mice were repeatedly injected 

with PBA (120 mg/kg BW, i.p.) every 3 h starting at 0.5 h 

after AAP challenge, since PBA demonstrates relatively rapid 

decay in vivo. Liver pathology was evaluated by H-E staining, 

and serum aminotransferases levels were determined. 

Oxidative stress was assessed by immunohistological staining 

for 4-hydroxynonenal (HNE). Total glutathione (GSH) content 

in the liver was measured colorimetrically. Apoptotic cell 

death was visualized by TdT-mediated dUTP nick end labeling 

(TUNEL). Phosphorylation of c-jun N—terminal kinase (p-JNK/ 

JNK) and cleavage of activating transcription factor (ATF) 6 

were detected by Western blotting. Results: A single injection 

of AAP caused massive necrosis of hepatocytes predominantly 

in pericentral area in 12 h as expected; however, subsequent 

repeated injections of PBA dramatically ameliorated these 

pathological changes in the liver. Indeed, the ALT levels were 

elevated to 11200 ± 455 IU/L at 6 h after AAP, whereas the 

values only reached 4510 ± 434 IU/L in mice treated with 

repeated PBA (p


showed enhanced mitochondrial translocation of JNK accompanied 

by an increase in the release of mitochondrial enzymes, 

such as apoptosis-inducing factor and endonuclease G, into 

the cytosol, which is indicative of increased mitochondrial dysfunction 

and subsequent nuclear DNA fragmentation. Finally, 

in vitro experiments showed that Gab1-deficient hepatocytes 

were more susceptible to APAP-induced mitochondrial dysfunction 

and cell death, suggesting that hepatocyte Gab1 is a 

direct target of APAP-induced hepatotoxicity. Conclusion: Our 

current data demonstrate that hepatocyte Gab1 plays a critical 

role in controlling the balance between hepatocyte death and 

compensatory hepatocyte proliferation during APAP-induced 

liver injury. Thus, hepatocyte Gab1 would be a potential therapeutic 

target for APAP-induced liver injury. 

Disclosures: 





The following authors have nothing to disclose: Yuichi Yoshida, Kunimaro Furuta, 

Satoshi Ogura, Tomohide Kurahashi, Mayumi Egawa, Shinichi Kiso, Yoshihiro 

Kamada 

228 

Lipotoxicity Accounts for Liver Injury due to Etomoxir: 

Application of Systems Toxicology Modeling of a Patient 

Population 

Scott Q. Siler 1,2 , Yuching Yang 2,1 , Diane M. Longo 2,1 , Brett A. 

Howell 2,1 , Paul Watkins 2 ; 1 DILI-sim, Castro Valley, CA; 2 Institute 

for Drug Safety Sciences, The Hamner Institutes-University of North 

Carolina, Research Triangle Park, NC 

Background: Etomoxir reduces fatty acid oxidation by inhibiting 

mitochondrial carnitine palmitoyltransferase 1 (CPT1). 

Etomoxir was originally developed for treatment of type 2 diabetes 

and congestive heart failure (CHF). In the “Etomoxir for 

the Recovery of Glucose Oxidation” (ERGO) clinical trial for 

CHF, elevations in serum ALT > 5 X ULN were observed in 

4 patients receiving active treatment, causing termination of 

the trial. It has remained unclear whether etomoxir hepatotoxicity 

resulted from direct mitochondrial toxicity or lipotoxicity 

due to the accumulation of lipids. Aim: To identify the 

most likely mechanistic determinant of etomoxir hepatotoxicity 

via simulations with DILIsym®. Methods: DILIsym®, a predictive, 

mechanistic, mathematical model of drug-induced liver 

injury (DILI), was used to simulate liver drug concentrations 

and DILI responses after administration of etomoxir (80 mg 

q.d.) in a virtual human population sample (SimPops) of 229 

simulated individuals. Quantitative fatty acid inhibition effects 

were based on in vitro data from the literature (Agius 1991). 

The DILIsym® model consists of various sub-models that are 

mathematically integrated to simulate patient response. The 

current work utilized several components of DILIsym®, including 

a physiologically-based pharmacokinetic (PBPK) sub-model 

representing drug distribution, sub-models of mitochondrial 

dysfunction and lipotoxicity, hepatocyte life cycle, and liver 

injury biomarkers. SimPops has been created by varying 

key parameters consistent with experimental data. DILIsym® 

is developed and maintained through the DILI-sim Initiative, 

a public-private partnership involving scientists in academia, 

industry, and the FDA. Results: In simulations where the model 

parameters excluded contributions of lipotoxicity to liver injury, 

no simulated patients were predicted to experience serum ALT 

increases. However, simulations including lipotoxicity showed 

an incidence of DILI that was similar to that seen in the ERGO 

trial, with 2-3% of simulated patients predicted to have ALT >5x 

ULN. The predicted ALT increases did not occur until after 4-6 

weeks of dosing, as was observed in the ERGO trial. Conclusions: 

The DILIsym® simulation results suggest that hepatotoxicity 

observed in the ERGO trial resulted from accumulation of 

lipids leading to lipotoxicity and not direct mitochondrial toxicity. 

This study illustrates the capability of DILIsym® to combine 

clinical data, in vitro data, predicted liver compound exposure, 

and inter-patient differences to provide insight into underlying 

mechanisms causing DILI. 

Disclosures: 

The following authors have nothing to disclose: Scott Q. Siler, Yuching Yang, 

Diane M. Longo, Brett A. Howell, Paul Watkins 

229 

HCV Core Gene Mutations with Heightened Liver Cancer 

Risk: Impact on Cellular Gene Expression 

Ahmed M. Elshamy 1 , Francis J. Eng 1 , Erin H. Doyle 1 , Arielle L. Klepper 

1 , Xiaochen Sun 1 , Angelo Sangiovanni 2 , Massimo Iavarone 2 , 

Massimo Colombo 2 , Robert E. Schwartz 3 , Yujin Hoshida 4 , Andrea 

D. Branch 1 ; 1 Liver Diseases, Icahn School of Medicine at Mount 

Sinai, New York, NY; 2 M. & A. Migliavacca Center for Liver Disease 

and 1st Division of Gastroenterology, Fondazione IRCCS 

Cà Granda Ospedale Maggiore Policlinico, University of Milan, 

Milan, Italy; 3 Department of Medicine, Weill Cornell Medical College, 

Department of Physiology, Biophysics, and Systems Biology, 

Weill Cornell Medical College, New York, NY; 4 Liver Cancer 

Program, Tisch Cancer Institute, Division of Liver Diseases, Department 

of Medicine, Icahn School of Medicine at Mount Sinai, New 

York, NY 

Background and Aims: Patients infected by genotype-1b hepatitis 

C virus (HCV) with Q 70 and/or M 91 core gene mutations 

have an almost five-fold increased risk of developing hepatocellular 

carcinoma (HCC) and an increased susceptibility to 

insulin resistance. The elevated HCC risk persists despite a sustained 

virological response to treatment. These compelling clinical 

data prompted us to seek laboratory data corroborating 

the oncogenic effects of the Q 70 /M 91 mutations. Methods: The 

HCC-associated virus (Q 70 /M 91 ) and the control virus (R 70 / 

L 91 ) were studied in Huh-7 cells. Infected cells were cultured 

in media containing adult human serum to promote differentiation. 

Quantitative RT-PCR and genome-wide transcriptome 

profiling (HumanHT12 beadarray, Illumina) were used for RNA 

analysis. Modulated molecular pathways were determined by 

Gene Set Enrichment Analysis. For clinical validation, transcriptomes 

of infected cells were compared to those of liver 

biopsies of patients with early-stage HCV-related cirrhosis who 

were followed prospectively for up to 23 yr to monitor for 

HCC and other clinical outcomes (n=216). Results: The Huh-7 

cells cultured in human serum acquired several features of differentiated 

hepatocytes, including a cobble-stone morphology 

and significantly enhanced expression of albumin and other 

hepatocyte-specific genes. Their expression of albumin was 

similar to that of cultured primary human hepatocytes and ~ 

20-fold higher than that of Huh-7 cells maintained in media 

containing fetal bovine serum. Surprisingly, although the two 

viruses were virtually identical in virological fitness, they had 

markedly different effects on cellular gene expression. The highrisk 

virus (Q 70 /M 91 ) enhanced expression of pathways associated 

with cancer and type II diabetes, while the control virus 

(R 70 /L 91 ) enhanced pathways associated with oxidative phosphorylation. 

Of special clinical relevance, the transcriptome of 

cells replicating the high-risk virus correlated significantly with 

an HCC-associated transcriptome in patients (Bonferroni-corrected 

P=0.03), but not with any other outcome-related transcriptome. 

The transcriptome of cells replicating the control 

virus did not correlate with any outcome-related transcriptome.


Conclusions: These results provide direct experimental evidence 

of sequence-specific oncogenic effects of HCV, recapitulating 

epidemiologic data and emphasizing the link between HCV 

sequences and clinical outcomes. These findings justify the 

assessment of HCV mutations as prognostic indicators. This 

simple cell-based system may be useful for investigating mechanisms 

of hepatocarcinogenesis and potential interventions 

(DA031095, DK090317). 

Disclosures: 

Massimo Colombo - Advisory Committees or Review Panels: BRISTOL-MEY- 

ERS-SQUIBB, SCHERING-PLOUGH, ROCHE, GILEAD, BRISTOL-MEYERS-SQUIBB, 

SCHERING-PLOUGH, ROCHE, GILEAD, Janssen Cilag, Achillion; Grant/ 

Research Support: BRISTOL-MEYERS-SQUIBB, ROCHE, GILEAD, BRISTOL-MEY- 

ERS-SQUIBB, ROCHE, GILEAD; Speaking and Teaching: Glaxo Smith-Kline, 

BRISTOL-MEYERS-SQUIBB, SCHERING-PLOUGH, ROCHE, NOVARTIS, GILEAD, 

VERTEX, Glaxo Smith-Kline, BRISTOL-MEYERS-SQUIBB, SCHERING-PLOUGH, 

ROCHE, NOVARTIS, GILEAD, VERTEX, Sanofi 


The following authors have nothing to disclose: Ahmed M. Elshamy, Francis J. 

Eng, Erin H. Doyle, Arielle L. Klepper, Xiaochen Sun, Angelo Sangiovanni, Massimo 

Iavarone, Robert E. Schwartz, Yujin Hoshida 

230 

Voluntary exercise opposes accelerated hepatocarcinogenesis 

in obese, diabetic mice by improving metabolic 

regulation and fatty liver disease, not via AMPK/ 

mTORC1 

Evi Arfianti 1 , Vanessa Barn 1 , W. G. Haigh 2 , Matthew M. Yeh 3 , 

George N. Ioannou 2 , Narci C. Teoh 1 , Geoffrey C. Farrell 1 ; 1 Liver 

Research Group, Australian National University (ANU) Medical 

School at The Canberra Hospital, Canberra, ACT, Australia; 2 Division 

of Gastroenterology, University of Washington, Seattle, WA; 

3 Division of Pathology, University of Washington, Seattle, WA 

Background Obesity and diabetes are independent risk factors 

for HCC associated with non-alcoholic fatty liver disease 

(NAFLD), HCV and other disorders. Diethylnitrosamine 

(DEN)-induced hepatocarcinogenesis is enhanced in obese, 

diabetic Alms1 mutant (foz/foz) mice which develop NASH. 

Such accelerated hepatocarcinogenesis is associated with 

hyperinsulinemia, hyperglycemia, and Akt/mTORC1 activation, 

but rapamycin fails to prevent HCC. We investigated 

whether exercise sufficient to slow weight gain reduces growth 

of dysplastic hepatocytes and HCC development in mice 

genetically predisposed to obesity and diabetes. Methods 

Male foz/foz and wild-type (Wt) littermates were injected with 

DEN (10mg/kg i.p.) or vehicle (saline) at age 12 days. From 

4wks, they were randomly assigned to cages provided with an 

exercise wheel (EW) until age 12 or 24 wks, or without EW. 

Dysplastic hepatocytes were identified by GST-pi immunohistochemistry 

(IHC), protein expression by immunoblotting, gene 

expression by real-time PCR, glucose tolerance by i.p. glucose 

tolerance test, hepatic lipid content using HPLC. Results: foz/ 

foz and Wt mice provided with in-cage exercise wheels ran 

an average of 4 km/day. In association with such physical 

activity, foz/foz weight gain slowed to that of Wt until 12 

wks; weight gain then accelerated but body weight remained 

lower than non-exercising mice. At 12wks, there was a significant 

reduction in GST-pi positive preneoplastic hepatocytes in 

exercising vs sedentary foz/foz mice. At 24 wks, more obese 

foz/foz mice had HCCs than lean Wt littermates (67% vs. 0, 

P6 tumors; 41.7%/41.7%/16.7% vs 

9.1%/18.1%/72.7%, p


sion rate, and malignant macroscopic classification. Conclusion: 

CTGF is up-regulated through Ras/Mek/Erk pathway in 

HCC. CTGF promotes the progression of HCC and related with 

its malignant characteristics. CTGF could be a new therapeutic 

target against HCC. 

Disclosures: 






The following authors have nothing to disclose: Yuki Makino, Takahiro Kodama, 

Minoru Shigekawa, Yugo Kai, Yasutoshi Nozaki, Tasuku Nakabori, Yoshinobu 

Saito, Satoshi Tanaka, Ryotaro Sakamori, Naoki Hiramatsu, Tomohide Tatsumi 

232 

p62 is a Key Regulator for Initiation and Development 

of Hepatocellular carcinoma 

Atsushi Umemura 1,2 , Yoshito Itoh 1 , Maria T. Diaz-Meco 3 , Jorge 

Moscat 3 , Michael Karin 2 ; 1 Department of Molecular Gastroenterology 

and Hepatology, Graduate School of Medical Science, 

Kyoto Prefectural University of Medicine, Kyoto, Japan; 2 Laboratory 

of Gene Regulation and Signal Transduction, Department of 

Pharmacology, University of California San Diego, San Diego, 

CA; 3 Sanford-Burnham Medical Research Institute, San Diego, CA 

p62, a scaffold protein interacting with various 

signaling pathways, accumulates and forms aggregates 

frequently observed in chronic liver diseases including NASH 

(non-alcoholic steatohepatitis), ASH (alcoholic steatohepatitis), 

and their progressed conditions, liver cirrhosis and hepatocellular 

carcinoma (HCC). p62 has also been reported as a 

critical oncoprotein involved in tumorigenesis and the development 

of many types of cancer including lung, breast, and 

kidney, whereas the tumorigenic role of p62 in the liver still 

remains unknown. It is reported that loss of p62 suppresses 

adenoma development in the autophagy knockout liver, importantly, 

however, this liver does not develop HCC. 

To elucidate the impact of p62 on HCC initiation and development, 

we decided to use two different animal models. First off, 

we injected a chemical carcinogen diethylnitrosamine (DEN), 

which induces multiple HCCs 7 months later, into liver-specific 

p62 knockout mice (p62KO) and their control mice (CON) 

to evaluate HCC development. We then established liver-specific 

TSC1 (tuberous sclerosis complex1)/p62 double-knockout 

mice (DKO). It is reported that liver-specific TSC1 knockout 

mice (TSC1KO) develop multiple HCCs spontaneously around 

10-12 months old. This means that DKO mice are useful in 

analyzing the role of p62 in hepatocarcinogenesis . CON mice developed multiple HCCs 7 months 

after DEN injection, but p62KO mice showed much lower incidence 

and smaller size of DEN-induced HCCs. Loss of TSC1 in 

the liver induced multiple HCCs as expected, but surprisingly, 

DKO mice did not developed HCCs nor even small tumors. In 

addition, liver injury and fibrosis observed in TSC1KO mice 

were attenuated in DKO livers. Among a number of signaling 

pathways, which p62 may be involved, Nrf2 was clearly activated 

in the livers of both DEN-injected CON and TSC1KO 

mice, and this activation was diminished in DEN-injected 

p62KO and DKO mice, respectively. Here we 

show that p62 plays important roles in HCC initiation as well 

as its development using two different HCC models. The Nrf2 

signaling pathway seems to be the most crucial in both models. 

Furthermore, additional p62 ablation ameliorated liver injury 

and fibrosis in TSC1KO mice. p62 is a well-known regulator of 

Nrf2 via its interaction with keap1. Since Nrf2 activation was 

suppressed by deletion of p62, the p62/keap1/Nrf2 signaling 

axis may profoundly contribute to hepatocarcinogenesis. In 

conclusion, p62 has a strong oncogenic function in the liver 

and is a promising target for HCC therapy. 

Disclosures: 

Yoshito Itoh - Grant/Research Support: MSD KK, Bristol-Meyers Squibb, Dainippon 

Sumitomo Pharm. Co., Ltd., Otsuka Pharmaceutical Co., Chugai Pharm 

Co., Ltd, Mitsubish iTanabe Pharm. Co.,Ltd., Daiichi Sankyo Pharm. Co.,Ltd., 

Takeda Pharm. Co.,Ltd., AstraZeneca K.K.:, Eisai Co.,Pharm.Ltd, FUJIFILM Medical 

Co.,Ltd., Gelaed Sciences Co., GlaxoSmithKline 

The following authors have nothing to disclose: Atsushi Umemura, Maria T. Diaz- 

Meco, Jorge Moscat, Michael Karin 

233 

Sitagliptin, a Dipeptidyl Peptidase 4 inhibitor, Suppressed 

Tumor Progression with Down-regulation of Nrf 

Nuclear Expression in a Mouse Model of Non-alcoholic 

Steatohepatitis-related Hepatocellular Carcinoma 

Takumi Kawaguchi, Eitaro Taniguchi, Minoru Itou, Tetsuharu Oriishi, 

Takuji Torimura; Division of Gastroenterology, Department of 

Medicine, Kurume University School of Medicine, Kurume, Japan 

Background and Aims: Sitagliptin is an anti-diabetic agent classified 

as a dipeptidyl peptidase 4 (DPP4) inhibitor. Although 

sitagliptin is known to improve non-alcoholic steatohepatitis 

(NASH), the effects of sitagliptin on NASH-related hepatocellular 

carcinoma (HCC) remains unclear. In addition, it is still 

unclear whether sitagliptin affects Nrf, a transcription factor 

regulating tumor proliferation via reprogramming of glucose 

metabolism. The aims of this study are to investigate effects of 

sitagliptin on HCC progression and nuclear expression of Nrf 

in a mouse model of NASH-related HCC. Material and Methods: 

A mouse model of NASH-related HCC without genetic 

modification (STAM mice) was employed in this study. Eightweek 

old STAM mice were orally administrated with either 

sitagliptin (30 mg/kg/day: DPP4 group; n=7) or distilled 

water (CON group; n=7) for 10 weeks. Then, the incidence, 

number, and volume of HCC were evaluated by contrast-enhanced 

computed tomography and histopathological examination. 

Nuclear expression of Nrf in HCC and non-tumor tissues 

were evaluated by immunostaining and quantified by Image 

J (U. S. National Institutes of Health, Bethesda, MD). Results: 

All mice survived until the end of this study. No significant 

difference was seen in body weight and the amount of food 

intake between the DPP4 and CON groups during the course 

of this study. While, liver-to-body weight ratio was significantly 

lower in the DPP4 group than in the CON group (7.3±1.2% vs. 

11.0±1.8%, P


Disclosures: 

The following authors have nothing to disclose: Takumi Kawaguchi, Eitaro Taniguchi, 

Minoru Itou, Tetsuharu Oriishi, Takuji Torimura 

234 

Two-step forward genetic screens of transposon mutagenesis 

and pooled shRNA library identify novel tumor 

suppressor genes and potential therapeutic target in 

HCC 

Takahiro Kodama, Nancy Jenkins, Neal Copeland; Cancer Biology, 

Houston Methodist Research Institute, Houston, TX 

Background: The use of emerging sequencing and genomics 

technologies has recently identified many mutated and/or differentially 

expressed genes in HCC. However, it is still difficult 

to identify true driver genes for HCC because of the large number 

of passenger mutations and other gene-regulatory mechanism 

such as epigenetic regulation, SNP and non-coding RNA. 

To overcome this difficulty, we performed two-step forward 

genetic screens and aimed to discover novel tumor suppressor 

genes (TSGs) in HCC. Methods: A transposon mutagenesis 

screen was performed using mice that harbor the conditional 

SB system (loxP-STOP-loxP(LSL)-SB11 transposase and a T2/ 

Onc2 or T2/Onc3 concatemer) and Alb-cre with or without 

sensitizing mutation of HBs-Ag transgene (Liver-SB/HBV or Liver-SB/noHBV). 

To validate candidate TSGs in a high-throughput 

manner, we subsequently performed in vivo small-scale 

pooled shRNA library screen using mouse immortalized liver 

progenitor cells (iLPCs). Results: We collected 334 and 146 

tumors from the Liver-SB/HBV and Liver-SB/noHBV mice, 

respectively, and identified 3523 and 2177 candidate cancer 

genes, respectively. List of 1917 candidate genes common 

in two transposon screening were narrowed down into 250 

genes that had non-synonymous mutation or mRNA downregulation 

reported in human HCC. Fifteen lentiviral pooled shRNA 

libraries (50 shRNAs/pool) targeting these 250 genes were 

introduced into iLPCs individually and assessed for their effects 

on subcutaneous tumor growth of transduced iLPCs in Nude 

mice. Eleven pooled libraries showed significant increase in 

tumor formation rate of iLPCs compared to negative control 

shRNA (16.7% - 32.1% vs 4.8%, P


236 

Clinical and Metabolic Effects Associated With Weight 

Loss and Obeticholic Acid In Nonalcoholic Steatohepatitis 

(NASH) 

Bilal Hameed 1 , Norah Terrault 1 , Ryan M. Gill 1 , Rohit Loomba 2 , 

Naga P. Chalasani 3 , Jay H. Hoofnagle 4 , Mark L. Van Natta 5 ; 

1 UCSF (University of California San Francisco), San Francisco, CA; 

2 University of California San Diego, San Diego, CA; 3 Indiana Univeristy, 

Indianapolis, IN; 4 NIDDK, Bethesda, MD; 5 John Hopkins 

University, Baltimore, MD 

Background/Aims: In a 72-week, randomized controlled trial 

(FLINT), obeticholic acid (OCA) was superior to placebo in 

improving serum ALT levels and liver histology in 283 patients 

with NASH. OCA therapy also reduced weight (mean net loss 

= 2.3 kg). Because weight loss can improve hepatic histology, 

we have assessed the role of weight loss and OCA treatment 

in improving clinical and metabolic features of NASH. Methods: 

This secondary analysis was limited to 200 patients who 

underwent a baseline and end-of-treatment liver biopsy. Liver 

histology was graded using the standardized NASH CRN Scoring 

System for NASH activity. Weight loss was defined as ≥ 2 

kg loss by the end of treatment. Results: Weight loss occurred 

in 43 of 102 (42%) OCA vs 29 of 98 (30%) placebo-recipients 

(p=0.08). NAS improved more with OCA in patients 

who lost vs did not lose weight (-2.4 vs -1.2, p





Inc. 

The following authors have nothing to disclose: Kellie Young, Maria Aguilar, Taft 

Bhuket, Benny Liu, Robert J. Wong 

238 

Application of transient elastography and FibroMeter in 

patients with nonalcoholic fatty liver disease 

Vincent W. Wong 1 , Grace LH Wong 1 , Sally Shu 1 , Angel ML 

Chim 1 , Anthony W. Chan 2 , Paul C. Choi 2 , Henry Lik-Yuen Chan 1 ; 

1 Department of Medicine and Therapeutics, The Chinese University 

of Hong Kong, Hong Kong, China; 2 Department of Anatomical 

and Cellular Pathology, The Chinese University of Hong Kong, 

Hong Kong, China 

Background: Nonalcoholic fatty liver disease (NAFLD) has 

emerged as a significant cause of cirrhosis and hepatocellular 

carcinoma, but the majority of patients have relatively minor 

disease. Hence, initial non-invasive assessments are preferred. 

Liver stiffness measurement (LSM) by transient elastography 

may fail in obese NAFLD patients. We therefore evaluated 

the use of LSM and serum-based FibroMeter tests in NAFLD 

patients. Methods: Consecutive liver biopsies for NAFLD were 

evaluated using the Kleiner’s system. LSM and blood tests were 

performed one day prior to liver biopsy. LSM was considered 

reliable if 10 valid acquisitions were obtained and the interquartile 

range-to-median ratio of LSM was ≤0.3. Cutoff values 

for LSM (7.9 and 9.6 kPa for ≥F3 disease) and FibroMeter-NA- 

FLD (0.62 for ≥F3 disease) were based on prior publications. 

Results: 293 examinations were analyzed (age 50 years [SD 

10], 54% males, body mass index 27.7 kg/m 2 [SD 4.2]). 

The distribution of fibrosis stages was 38% F0, 30% F1, 9% 

F2, 11% F3 and 12% F4. FibroMeter score was valid in all 

patients. Among 233 patients undergoing LSM, 224 (96%) 

had 10 valid acquisitions and 203 (87%) had reliable measurements. 

In the overall population, the area under the receiver-operating 

characteristics curve (AUROC) for FibroMeter was 

0.72 (95% CI 0.66-0.78) for ≥F2, 0.73 (95% CI 0.67-0.80) 

for ≥F3 and 0.79 (95% CI 0.72-0.87) for F4. At the cutoff of 

0.62, the sensitivity, specificity, positive and negative predictive 

values of FibroMeter for ≥F3 disease were 25%, 91%, 

46% and 81%, respectively. In patients with reliable LSM, the 

corresponding AUROC was 0.83 (95% CI 0.77-0.89), 0.91 

(95% CI 0.87-0.95) and 0.91 (95% CI 0.86-0.97), respectively. 

The LSM cutoff of 7.9 kPa had 93% sensitivity and 98% 

negative predictive value in excluding ≥F3 disease. On the 

other hand, the cutoff of 9.6 kPa had 86% specificity and 

only 60% positive predictive value in ruling in ≥F3 disease. If 

FibroMeter was performed in 113 patients with unreliable LSM 

or LSM ≥7.9 kPa, 61 of 98 (62%) patients with a score


Norah Terrault - Advisory Committees or Review Panels: Eisai, Biotest; Consulting: 

BMS, Merck, Achillion; Grant/Research Support: Eisai, Biotest, Vertex, 

Gilead, AbbVie, Novartis, Merck 




Kris V. Kowdley - Advisory Committees or Review Panels: Achillion, BMS, Evidera, 

Gilead, Merck, Novartis, Trio Health, Abbvie; Grant/Research Support: 

Evidera, Gilead, Immuron, Intercept, Tobira; Speaking and Teaching: Abbvie, 

Gilead 



Scholar Rock 

Reshma Shringarpure - Employment: Intercept 






The following authors have nothing to disclose: Arthur J. McCullough, Saswati 

Hazra, Xiaohong Yan, Leigh MacConell 

240 

Omega-3 fatty acids improve proteomic and lipidomic 

markers of endoplasmic reticulum (ER) stress and mitochondrial 

dysfunction in a randomized controlled trial in 

subjects with Nonalcoholic Steatohepatitis 

Livia Rodrigues 1 , Claudia P. Oliveira 1 , José Tadeu Stefano 1 , 

Monize A. Nogueira 1 , Ismael D. Silva 2 , Edson G. Lo Turco 3 , 

Venancio Avancini F. Alves 4 , Flair J. Carrilho 1 , Puneet Puri 5 , Dan 

Waitzberg 1 ; 1 Gastroenterology, University of São Paulo School of 

Medicine, São Paulo, Brazil; 2 Gynecology, Universidade Federal 

de São Paulo, São Paulo, Brazil; 3 Surgery/Urology, Universidade 

Federal de São Paulo, São Paulo, Brazil; 4 Patology, University of 

São Paulo School of Medicine, São Paulo, Brazil; 5 Gastroenterology, 

Hepatology and Nutrition, Virginia Commonwealth University-VCU, 

Richmond, VA 

Background/Aims: There is no effective FDA approved treatment 

for nonalcoholic steatohepatitis (NASH). Increased 

n-6/n-3 polyunsaturated fatty acid (PUFA) ratio can induce 

endoplasmic reticulum (ER) stress and mitochondrial dysfunction 

that characterize NASH. We hypothesized that n-3 PUFA supplementation 

would improve these abnormalities. We aimed to 

define the hepatic proteomic and plasma lipidomic profiles following 

n-3 PUFA therapy in subjects with NASH and relate it to 

markers of ER stress and mitochondrial dysfunction. Methods: 

A 6-month double blind randomized controlled trial in subjects 

with biopsy proven NASH was conducted with n-3 PUFA (945 

mg [α linolenic acid/ 64%, eicosapentaenoic acid (EPA)/16% 

and docosahexaenoic acid (DHA)/21%]), 3 capsules/day and 

matched placebo. A 6-month follow up liver biopsy was performed 

per IRB protocol. Hepatic proteomics from formalin-fixed 

paraffin embedded liver tissue and plasma lipidomics were 

performed at baseline and 6-months in the n-3 PUFA group 

using mass spectrometry. The proteins and lipids were matched 

to UniProt and LIPID MAPS database respectively. Cytoscape 

software was used to analyze functional pathways. Results: 

Age and gender matched 60 NASH subjects (n=32, n-3 PUFA 

treatment group; n=28, placebo group) completed the study. A 

6-month n-3 PUFA therapy significantly (all p 2x baseline and > 5x 

ULN, occurring within the first 24 weeks of treatment were 

evaluated in context of subsequent clinical outcomes through 

Week 48. Results: Overall, 29/172, 25/170, 3/174 and 

25/163 patients, respectively, from Arms A-D met criteria for 

ALT flare; of those, 18, 16, 2, and 14 patients were baseline 

HBeAg-positive. A greater proportion of patients in Arms 

A, B and D who experienced ALT flares achieved subsequent 

HBeAg loss, HBsAg decline ≥ 1log 10 

IU/ml, or HBsAg loss 

compared to those who did not experience an ALT flare (table).


Comparing two combination therapies, Arm A achieved higher 

rates of HBsAg loss (P=0.016) following flares than Arm B. 

This is partly driven by HBeAg-negative patients in Arm A who 

attained disproportionately increasing rates of HBsAg loss and 

HBsAg decline ≥ 1log 10 

IU/ml which was not observed in 

Arm B. Conclusion: Treatment-associated ALT flares, especially 

with TDF+PEG combination therapy, are associated with clinical 

endpoints related to HBV immune control. ALT flares were 

higher in the TDF+PEG x48w group and associated with the 

highest HBsAg loss. 

Table: Proportion of Patients who Achieved Clinical Endpoints by 

ALT flare status (N=679) 

*Includes subjects with baseline HBeAg-positive only. 

Disclosures: 

Henry Lik-Yuen Chan - Advisory Committees or Review Panels: Gilead, MSD, 

Bristol-Myers Squibb, Roche, Novartis Pharmaceutical, Abbvie; Speaking and 

Teaching: Echosens 







Phillip Dinh - Employment: Gilead Sciences; Stock Shareholder: Gilead Sciences 

Eduardo B. Martins - Employment: Gilead Sciences, Inc.; Stock Shareholder: 

Gilead Sciences, Inc. 

Leland J. Yee - Employment: Gilead Science 

Prista Charuworn - Stock Shareholder: Gilead Sciences 

Mani Subramanian - Employment: Gilead Sciences 






Seng Gee Lim - Advisory Committees or Review Panels: Bristol-Myers Squibb, 

Novartis Pharmaceuticals, Merck Sharp and Dohme Pharmaceuticals, Gilead 

Pharmaceuticals, Roche Pharmaceuticals; Speaking and Teaching: Bristol-Myers 

Squibb, Novartis Pharmaceuticals, Merck Sharp and Dohme Pharmaceuticals, 

Gilead Pharmaceuticals 

Scott Fung - Advisory Committees or Review Panels: Gilead Sciences, Merck, 

Vertex, Roche; Speaking and Teaching: BMS, Gilead, AbbVie, Janssen, Merck 







Maria Buti - Advisory Committees or Review Panels: Gilead, Janssen, MSD; 

Grant/Research Support: Gilead, Janssen; Speaking and Teaching: Gilead, 

Janssen, BMS 

Giovanni B. Gaeta - Advisory Committees or Review Panels: Janssen, Merck, 

Abbvie, Roche; Speaking and Teaching: BMS, Gilead 

George V. Papatheodoridis - Advisory Committees or Review Panels: Merck 

Sharp & Dohme, Novartis, Abbvie, Boerhinger Ingelheim, Bristol-Meyer Squibb, 

Gilead, Roche, Janssen, GlaxoSmith Kleine; Grant/Research Support: Roche, 

Gilead, Bristol-Meyer Squibb, Abbvie, Janssen; Speaking and Teaching: Merck 

Sharp & Dohme, Bristol-Meyer Squibb, Gilead, Roche, Janssen, Abbvie 

Robert Flisiak - Advisory Committees or Review Panels: Gilead, Bristol Myers 

Squibb, Janssen, Merck, Roche, Abbvie; Consulting: Janssen, Bristol Myers 

Squibb, Gilead, Merck, Abbvie; Grant/Research Support: Roche; Speaking and 

Teaching: Janssen, Roche, Bristol Myers Squibb, Gilead, Merck, Abbvie 

Patrick Marcellin - Consulting: Roche, Gilead, BMS, Vertex, Novartis, Janssen, 

MSD, Abbvie, Alios BioPharma, Idenix, Akron; Grant/Research Support: Roche, 

Gilead, BMS, Novartis, Janssen, MSD, Alios BioPharma; Speaking and Teaching: 

Roche, Gilead, BMS, Vertex, Novartis, Janssen, MSD, Boehringer, Pfizer, 

Abbvie 

The following authors have nothing to disclose: Sang Hoon Ahn, Fehmi Tabak, 

Rajiv Mehta, Kyungpil Kim, Aric Josun Hui 

242 

Nucleos(t)ide Analogues against Hepatitis B Virus 

Reduces the Risk of Various Major Malignancies: a 

Nationwide Cohort Study Based on the Health Insurance 

Review and Assessment Service Database. 

Donghyeon Lee 1 , Jeong-Hoon Lee 1 , Yuri Cho 1 , Jeong-Ju Yoo 1 , 

Minjong Lee 1 , Young Youn Cho 1 , Hyeki Cho 1 , Hongkeun Ahn 1 , 

June Sung Lee 2 , Eun Ju Cho 1 , Su Jong Yu 1 , Yoon Jun Kim 1 , Jung- 

Hwan Yoon 1 ; 1 Seoul national university hospital, Seoul, Korea (the 

Republic of); 2 Inje University, Ilsan Paik Hospital, Ilsan, Korea (the 

Republic of) 

Background: Preceding epidemiology studies reported that 

chronic hepatitis B virus (HBV) infection is a risk factor for various 

malignancies including hepatocellular carcinoma (HCC), 

cholangiocarcinoma, non-Hodgkin lymphoma, and stomach 

cancer. These associations have been partially explained by 

the HBV-inducing immune tolerance to malignant cells. It has 

been well proven that antiviral treatment in chronic hepatitis 

B (CHB) patients reduces the risk of HCC. However, it is still 

unclear whether nucleos(t)ide analogues (NAs) against HBV 

could reduce the risk of various malignancies except for HCC. 

In this study, we evaluated the association of anti-HBV treatment 

with NAs and the risk of various cancers in CHB patients. 

Methods: The National Claims Database from Health Insurance 

Review and Assessment Service (HIRA) was used. From 

this database, we included CHB patients who received anti- 

HBV NAs (i.e., lamivudine, telvibudine, clevudine, adefovir, 

entecavir, and tenofovir) for > 30 days between 2009 and 

2013 (the NA group) or no antiviral treatment (the control 

group). Date of data cutoff was December 31, 2014. The 

hazard ratio (HR) for NAs was calculated by Cox regression 

model. Results: A total of 299,740 patients with CHB were 

included and 39,900 patients (13.3%) were involved in the 

NA group. During the study period 14,186 patients developed 

any kind of malignancy. In multivariate analysis, the NA 

group showed significantly lower risk of developing new malignancies 

(vs the control group; HR=0.511, 95% confidence 

interval=0.466-0.561, P


243 

Biochemical, Virologic, and Serologic Response Following 

Discontinuation of Long Term TDF Therapy 

Maria Buti 1 , David K. Wong 2 , Edward J. Gane 3 , Robert Flisiak 4 , 

Michael P. Manns 5 , Kelly D. Kaita 6 , Lanjia Lin 7 , Kathryn M. Kitrinos 

7 , John F. Flaherty 7 , Anuj Gaggar 7 , Patrick Marcellin 8 , Harry 

L. Janssen 9 ; 1 Hospital General Universitari Vall d’Hebron and 

Ciberehd, Barcelona, Spain; 2 University of Toronto, Toronto, ON, 

Canada; 3 Auckland City Hospital, Auckland, New Zealand; 4 Medical 

University of Bialystok, Bialystok, Poland; 5 Medical School of 

Hannover, Hannover, Germany; 6 University of Manitoba, Winnipeg, 

MB, Canada; 7 Gilead Sciences, Inc., Foster City, CA; 8 Hôpital 

Beaujon, Clichy, France; 9 ErasmusMC, Rotterdam, Netherlands 

Introduction: Treatment of HBV with tenofovir disoproxil fumarate 

(TDF) is associated with durable viral suppression, liver 

fibrosis improvement, and no documented resistance in a 

cohort of patients followed for up to 8 years. We characterized 

the responses following treatment discontinuation. Methods: 

Patients from 2 phase 3 studies of TDF in HBeAg-negative and 

HBeAg-positive patients were followed for up to 24 weeks after 

discontinuing TDF. Clinical and laboratory parameters, and 

pre-specified requirements to restart therapy were monitored 

during the treatment-free follow-up (TFFU) period. Results: To 

date, 123 patients entered TFFU with mean duration of 12 

weeks and 71 patients completing 24 weeks of TFFU. 98 (80%) 

patients were HBeAg negative at the time of TDF withdrawal, 

including 21 that seroconverted in the parent study. Serious 

adverse events (AEs) were reported in 10 subjects (8%), 7 

of which involved ALT elevation, and Grade 3-4 AEs were 

observed in 12% of subjects. 89% of patients had increase in 

HBV DNA to >69 IU/mL and 28% had Grade 3-4 elevations in 

ALT after cessation of TDF. At last TFFU visit, 44 patients (36%) 

had normal ALT, 59 (48%) maintained HBV DNA


were younger (49.9 vs. 537; P90%. 

Conclusions: The results demonstrate that serum HBV RNA levels 

could be used early during PegIFNα-2a therapy, and that 

they appear superior to conventional biomarkers for predicting 

non-response in HBeAg-positive patients. 

Biomarker Cutoff Performance Characteristics at Treatment Week 

12 

Disclosures: 

Florian van Bömmel - Advisory Committees or Review Panels: Gilead Sciences 

Inc., Roche Pharmaceutics; Grant/Research Support: Biopredictive; Speaking 

and Teaching: Bristol-Myers Squibb, Janssen, Roche Pharmaceutics, Gilead Sciences 

Inc., Abbvie 

Hua He - Employment: Roche 

Cynthia Wat - Employment: Roche Products Ltd 

Vedran Pavlovic - Employment: Roche Products Ltd; Stock Shareholder: Roche 

Products Ltd 

Lei Yang - Employment: Roche (China) Holding Ltd. 







The following authors have nothing to disclose: Alena van Bömmel, Alexander 

Krauel, Danilo Deichsel, Stephan Boehm 

246 

The Predictive Values of Hepatocellular Carcinoma Risk 

Scores are not Affected from the Antiviral Therapy in 

Patients with Chronic Hepatitis B Infection 

Ulus S. Akarca, Fulya Gunsar, Ilker Turan, Galip Ersoz, Zeki 

Karasu, Omer Ozutemiz; Division of Gastroenterology, Ege University 

Faculty of Medicine, Izmir, Turkey 

Background and Aims: Hepatocellular carcinoma (HCC) 

remains the major complication of chronic hepatitis B (CHB). 

Several scoring systems including CU-HCC, GAG-HCC, and 

REACH-B have been developed to assess the risk for development 

of HCC. In addition to the patients who do not receive 

antiviral treatment, the predictive value of these scores for HCC 

was found to be good in patients on treatment. In the present 

study, we aimed to assess the impact of antiviral therapy for the 

predictive values of HCC risk scores in Caucasian CHB patients 

and compare the predictive values in patients on treatment and 

without treatment. Methods: We investigated the predictive 

values of CU-HCC, GAG-HCC, and REACH-B scores in Caucasian 

HBsAg-positive patients who had baseline labs including 

HBV DNA at presentation. Patients who had (1) HCC at presentation; 

(2) HCV, HDV and HIV coinfection; (3) missing data 

for the calculation of the scores were excluded. Results: We 

analyzed 2331 patients (M/F:1370/961, mean age 42±13). 

In this study group, 623 patients were inactive HBV carrier, 

1454 patients were CHB, 163 patients had cirrhosis and 91 

patients had HBsAg seroconversion during follow-up. HCC 

developed in 37 patients during median 4.9 year follow-up.


Three hundred forty-one patients were on lamivudine, 60 were 

on adefovir, 188 were on entecavir, 467 were on tenofovir, 

and 32 were on combination therapies. ROC analysis for the 

predictive values of HCC scores showed that AUROC values 

were 0.804 for CU-HCC, 0.809 for REACH-B and 0.882 for 

GAG-HCC. These values were similar in patients without therapy 

and with entecavir and tenofovir therapies as in the literature. 

The most predictive GAG-HCC score had sensitivity of 

97.3% and negative predictive value of 99.9%. Conclusion: 

In this large cohort of Caucasian HBV patients, all of these 

scores had good predictive values for HCC development both 

in patients on treatment and in patients who did not receive 

antiviral treatment. 

Disclosures: 

Ulus S. Akarca - Advisory Committees or Review Panels: GILEAD, BMS, MSD, 

AbbVie 

The following authors have nothing to disclose: Fulya Gunsar, Ilker Turan, Galip 

Ersoz, Zeki Karasu, Omer Ozutemiz 

247 

High Rates of SVR in Treatment-Experienced Patients 

with Genotype 1 HCV Infection and Cirrhosis After 

Treatment with Ledipasvir/Sofosbuvir and Vedroprevir 

with or Without Ribavirin for 8 weeks 

Eric Lawitz 1 , Fred Poordad 1 , Robert H. Hyland 2 , Lin Liu 2 , Hadas S. 

Dvory 2 , Phillip S. Pang 2 , Diana M. Brainard 2 , Julio A. Gutierrez 1 ; 

1 Texas Liver Institute, San Antonio, TX; 2 Gilead Sciences, Inc, Foster 

City, CA 

Background: Ledipasvir/sofosbuvir (LDV/SOF) fixed dose combination 

± ribavirin (RBV) for 12 weeks in patients with HCV 

genotype 1 and cirrhosis is safe and effective. We previously 

showed that LDV/SOF plus the non-nucleotide NS5B inhibitor 

GS-9669 for 8 weeks resulted in 82-91% SVR12 in patients 

with cirrhosis and genotype 1 HCV infection. In the current 

study, we evaluated the safety and efficacy of 8 weeks of LDV/ 

SOF plus the HCV NS3 protease inhibitor vedroprevir (VDV, 

GS-9451) ± RBV in genotype 1 HCV-infected patients with 

cirrhosis. Methods: Patients, previously treated with pegylated 

interferon+RBV, with compensated cirrhosis and chronic HCV 

genotype 1 infection were randomized equally to receive LDV/ 

SOF+VDV or LDV/SOF+VDV+RBV for 8 weeks. Deep sequencing 

was performed for all patients who didn’t achieved SVR12. 

Results: 46 patients were randomized and treated. The majority 

were male (65%), Caucasian (96%), and had HCV genotype 

1a (70%), and were IL28B non-CC (87%). The median 

HCV RNA was 6.0 log 10 

IU/ml. Rates of on-treatment Week 4 

viral suppression, SVR12, and relapse are shown in the table. 

NS5A RAVs were detected in all 3 subjects who relapsed. 

LDV/SOF+VDV±RBV was safe and well tolerated. There were 

no SAEs. One patient was lost to follow up having completed 

only 4 weeks of therapy. There were no discontinuations due 

to adverse events. Adverse events were generally mild, and 

Grade 3/4 laboratory abnormalities were infrequent. The most 

frequent adverse events were headache, rash, and sinusitis. 

Conclusions: Ledipasvir/sofosbuvir +VDV for 8 weeks achieved 

high SVR rates in genotype 1 HCV-infected treatment-experienced 

patients with cirrhosis irrespective of the addition of RBV. 

Both regimens were safe and well tolerated. 

Disclosures: 













Robert H. Hyland - Employment: Gilead Sciences, Inc; Stock Shareholder: Gilead 

Sciences, Inc 

Lin Liu - Employment: Gilead Sciences, Inc. 



Sciences 

The following authors have nothing to disclose: Hadas S. Dvory, Julio A. Gutierrez 

248 

High SVR4 Rates Achieved With the Next Generation 

NS3/4A Protease Inhibitor ABT-493 and NS5A Inhibitor 

ABT-530 in Non-Cirrhotic Treatment-Naïve and Treatment-Experienced 

Patients With HCV Genotype 3 Infection 

(SURVEYOR-2) 

Paul Y. Kwo 1 , Michael Bennett 2,3 , Stanley Wang 4 , Hugo E. 

Vargas 5 , David L. Wyles 6 , J. Scott Overcash 7 , Peter J. Ruane 8 , 

Benedict Maliakkal 9 , Asma Siddique 10 , Bal R. Bhandari 11 , Fred 

Poordad 12 , Ran Liu 4 , Chih-Wei Lin 4 , Teresa Ng 4 , Federico J. 

Mensa 4 , Jens Kort 4 ; 1 Division of Gastroenterology, Indiana University 

Medical Center, Indianapolis, IN; 2 San Diego Digestive 

Disease Consultants, Inc., San Diego, CA; 3 Medical Associates 

Research Group, Inc., San Diego, CA; 4 AbbVie Inc., North Chicago, 

IL; 5 Mayo Clinic, Phoenix, AZ; 6 University of California San 

Diego, La Jolla, CA; 7 eStudySite, San Diego, CA; 8 Ruane Medical 

& Liver Health Institute, Los Angeles, CA; 9 University of Rochester 

Medical Center, Rochester, NY; 10 Virginia Mason Hospital and 

Medical Center, Seattle, WA; 11 Gastroenterology & Nutritional 

Medical Services, Bastrop, LA; 12 Texas Liver Institute, University of 

Texas Health Science Center, San Antonio, TX 

Background: The next generation HCV direct acting antivirals 



characterized by potent pan-genotypic in vitro antiviral activity 

and a high barrier to resistance selection with retention 

of activity against key known resistance-associated variants. 

ABT-493 and ABT-530 each provided a mean 4 log 10 

IU/mL 

decline from baseline in HCV plasma viral load after 3 day 

monotherapy in genotype (GT) 1-infected subjects and have 

comparable in vitro antiviral potency against GT3a. Purpose: 

To evaluate the efficacy and safety of ABT-493 and ABT-530 

with or without ribavirin (RBV) in non-cirrhotic GT3-infected 

treatment-naïve (TN) and pegylated interferon/RBV (pegIFN/ 

RBV) treatment experienced (TE) subjects Methods: Subjects 

received 12 weeks of ABT-493 300 mg+ABT-530 120 mg 

(Arm D), ABT-493 200 mg+ABT-530 120 mg (Arm E), ABT- 

493 200 mg+ABT-530 120 mg+RBV (Arm F), or ABT-493 200 

mg+ABT-530 40 mg (Arm G). DAAs were dosed once daily; 

weight-based RBV (1000 or 1200 mg) was dosed twice daily. 

The primary efficacy endpoint is sustained virologic response 

12 weeks after the last dose of study drug (SVR12); we present 

post-treatment week 4 data (SVR4) here. Safety was evaluated 

by monitoring adverse events (AEs), laboratory tests, and vital 

signs. Results: 120 GT3-infected subjects were treated in Arms


D (n=30), E (n=29), F (n=31), or G (n=30). Subjects were 

male, 56%; median age, 52.0 years; GT3a, 98%; TN, 92%; 

TE, 8%; fibrosis >F2, 15%; median baseline HCV RNA log 10 

IU/mL, 6.7. There has been 1 virologic failure thus far in each 

treatment arm (n=4), 3 of which were in pegIFN/RBV TE subjects. 

One subject in Arm G was lost to follow-up at the week 

2 visit. The SVR4 rate was 96% (27/28), 96% (27/28), 97% 

(29/30) and 93% (27/29) in Arms D, E, F, and G, respectively, 

among subjects with available post-treatment Week 4 

data. AEs were mostly mild, with most common DAA-related 

AEs being fatigue, nausea, and headache. There were no serious 

DAA-related AEs; 1 subject discontinued due to DAA- and 

RBV-related AEs of abdominal pain and heat sensation. Typical 

reductions in hemoglobin were observed in the RBV containing 

arm (Arm F). Conclusions: ABT-493+ABT-530 treatment for 

12 weeks with or without RBV in TN or TE noncirrhotic HCV 

GT3-infected subjects was well tolerated. Promising SVR4 rates 

of 93%–96% were achieved without RBV. SVR12 data and 

further details regarding virologic failures will be available 

at the time of the meeting. These encouraging results warrant 

further study of this once-daily ABT-493/ABT-530 regimen in 

GT3 patients, including patients with cirrhosis. 

Disclosures: 





Stanley Wang - Employment: AbbVie 

David L. Wyles - Advisory Committees or Review Panels: Bristol Myers Squibb, 

Janssen, Merck; Grant/Research Support: Gilead, Merck, Bristol Myers Squibb, 

AbbVie, Tacere 

Peter J. Ruane - Advisory Committees or Review Panels: BMS; Consulting: Gilead, 

Abbvie, Janssen; Grant/Research Support: BMS, Gilead, Merck, Abbvie, Idenix, 

Idenix, Janssen, Viiv; Speaking and Teaching: Gilead, Merck, Abbvie, Abbvie, 

Janssen; Stock Shareholder: Gilead, Gilead 

Benedict Maliakkal - Speaking and Teaching: Valeant Pharma (Salix), Gilead, 

AbbVie, Bristol Myers Squibb 









AbbVie 



The following authors have nothing to disclose: Michael Bennett, Hugo E. Vargas, 

J. Scott Overcash, Asma Siddique, Bal R. Bhandari 

249 

Sofosbuvir/GS-5816 Fixed Dose Combination for 12 

Weeks Compared to Sofosbuvir with Ribavirin for 24 

Weeks in Genotype 3 HCV Infected Patients: The Randomized 

Controlled Phase 3 ASTRAL-3 Study 

Alessandra Mangia 1 , Stuart K. Roberts 2 , Stephen Pianko 3 , Alex 

J. Thompson 4 , Curtis Cooper 5 , Brian Conway 6 , Marc Bourlière 

7 , Tarik Asselah 8 , Thomas Berg 9 , Stefan Zeuzem 10 , William 

M. Rosenberg 11 , Kosh Agarwal 12 , Edward J. Gane 13 , Catherine 

A. Stedman 14 , Francesco Mazzotta 15 , Tram T. Tran 16 , Stuart 

C. Gordon 17 , Evguenia S. Svarovskaia 18 , Lingling Han 18 , 

John McNally 18 , Anu Osinusi 18 , Diana M. Brainard 18 , John G. 

McHutchison 18 , Nezam H. Afdhal 19 , Graham R. Foster 20 ; 1 Casa 

Sollievo della Sofferenza Hospital, San Giovanni Rotondo, Italy; 

2 Alfred Health and Monash University, Melbourne, VIC, Australia; 

3 Monash Health and Monash University, Melbourne, VIC, Australia; 

4 St Vincent’s Hospital, Melbourne, VIC, Australia; 5 Ottawa 

General Hospital, Ottawa, ON, Canada; 6 Vancouver Infectious 

Diseases Centre, Vancouver, BC, Canada; 7 Hôpital Saint Joseph, 

Marseilles, France; 8 University Paris Diderot, INSERM U773, Paris, 

France; 9 University Hospital Leipzig, Leipzig, Germany; 10 Johann 

Wolfgang Goethe University, Frankfurt, Germany; 11 University 

College London, London, United Kingdom; 12 Kings College 

Hospital, London, United Kingdom; 13 Auckland Clinical Studies, 

Auckland, New Zealand; 14 Christchurch Clinical Studies Trust and 

University of Otago, Christchurch, New Zealand; 15 Santa Maria 

Annunziata Hospital, Florence, Italy; 16 Cedars-Sinai Medical Center, 

Los Angeles, CA; 17 Henry Ford Health System, Detroit, MI; 

18 Gilead Sciences, Inc., Foster City, CA; 19 Beth Israel Deaconess 

Medical Center, Boston, MA; 20 Queen Mary University of London, 

London, United Kingdom 

Introduction: GS-5816 is a pangenotypic HCV NS5A inhibitor. 

In Phase 2 studies, the combination of sofosbuvir (SOF) and 

GS-5816 for 12 weeks demonstrated high efficacy in patients 

with genotype 3 HCV. This Phase 3 study compared treatment 

with a fixed dose combination (FDC) of SOF/GS-5816 for 

12 weeks to standard of care, SOF+RBV for 24 weeks, in 

patients with genotype 3 HCV. Methods: Patients at 75 sites 

in North America, Europe, Australia and New Zealand were 

randomized 1:1 to received SOF/GS-5816 (400 mg /100 

mg daily) FDC for 12 weeks or SOF (400mg daily) with RBV 

(1000-1200mg daily) for 24 weeks. HCV RNA was measured 

with the CAP/CTM HCV 2.0 assay with LLOQ =15 IU/mL The 

primary endpoint was sustained virologic response 12 weeks 

after treatment (SVR12) with a pre-specified non-inferiority margin 

of 10%. Results: Of 552 patients randomized and treated, 

62% were male, 89% were white, 39% had IL28B CC genotype, 

26% had prior treatment failure, and 30% had cirrhosis. 

Eight patients discontinued treatment due to adverse events, 

all of which were in the SOF +RBV treatment group. The most 

common AEs (>10% in either treatment group) are presented 

below. Five patients in the SOF/GS-5816 treatment group 

and 8 patients in the SOF+RBV treatment group experienced 

SAEs. One SAE, acute generalized exanthematous pustulosis, 

in a SOF+RBV treated patient was assessed as related to study 

drugs by the investigator. Hemoglobin decline and total bilirubin 

increases were more commonly observed in the group 

treated with SOF +RBV consistent with RBV–induced hemolysis. 

No other significant lab abnormalities were observed. HCV 

RNA declined rapidly in both treatment groups with 92% and 

88% of patients achieving HCV RNA < LLOQ after 4 weeks 

of treatment in the SOF/GS-5816 and SOF+RBV treatment 

groups, respectively. SVR12 data for all patients will be presented. 

Conclusions: The once daily, all-oral, single tablet regimen 

of SOF/GS-5816 was well tolerated in treatment-naïve 

and treatment-experienced genotype 3 HCV-infected patients


with and without cirrhosis. There were no discontinuations due 

to adverse events and a lower incidence of fatigue, insomnia 

and irritability in patients treated with SOF/GS-5816 for 

12 weeks compared to patients treated with SOF+RBV for 24 

weeks. 











The following authors have nothing to disclose: Francesco Mazzotta, Lingling 

Han 

Disclosures: 

Alessandra Mangia - Advisory Committees or Review Panels: ROCHE, Janssen, 

MSD, ROCHE, Janssen, MSD, Boheringer ; Consulting: Gilead; Grant/Research 

Support: Shering-Plough, Shering-Plough 


Stephen Pianko - Advisory Committees or Review Panels: Roche, Novartis, GIL- 

EAD, Roche, Novartis; Consulting: GILEAD; Speaking and Teaching: JANSSEN 




Abbvie, BMS 

Curtis Cooper - Advisory Committees or Review Panels: Gilead, Abbvie, MERCK; 

Grant/Research Support: MERCK, Gilead, Abbvie; Speaking and Teaching: 

MERCK, Abbvie, Gilead 










Squibb 

Tarik Asselah - Advisory Committees or Review Panels: AbbVie, Merck, Gilead, 

BMS, Roche, Janssen 









William M. Rosenberg - Advisory Committees or Review Panels: Janssen, Merk, 

Gilead, Merk, Gilead, GSK; Board Membership: iQur Limited, iQur Limited; 

Consulting: siemens; Speaking and Teaching: siemens, Roche 

Kosh Agarwal - Advisory Committees or Review Panels: Gilead, BMS, Novartis, 

Janssen, AbbVie, Gilead; Consulting: MSD, Janssen; Grant/Research Support: 

Roche, Gilead, BMS, BMS; Speaking and Teaching: Astellas 




Catherine A. Stedman - Advisory Committees or Review Panels: MSD, Abbvie; 


Tram T. Tran - Advisory Committees or Review Panels: Gilead, Bristol Myers 

Squibb; Consulting: Gilead, AbbVie, Janssen; Grant/Research Support: Bristol 

Myers Squibb; Speaking and Teaching: Gilead 






John McNally - Employment: Gilead Sciences, Inc; Stock Shareholder: Gilead 

Sciences, Inc 



Sciences 


Sciences 

250 

High SVR4 Rates Achieved With the Next Generation 

NS3/4A Protease Inhibitor ABT-493 and NS5A Inhibitor 

ABT-530 in Non-Cirrhotic Treatment-Naïve and Treatment-Experienced 

Patients With HCV Genotype 2 Infection 

(SURVEYOR-2) 

David L. Wyles 2 , Mark S. Sulkowski 3 , Stanley Wang 1 , Michael 

Bennett 4 , Hugo E. Vargas 5 , J. Scott Overcash 6 , Benedict Maliakkal 

7 , Asma Siddique 8 , Bal R. Bhandari 9 , Fred Poordad 10 , Sandra 

S. Lovell 1 , Chih-Wei Lin 1 , Teresa Ng 1 , Federico J. Mensa 1 , Jens 

Kort 1 ; 1 AbbVie, Inc., North Chicago, IL; 2 University of California 

San Diego, La Jolla, CA; 3 Johns Hopkins University School of Medicine, 

Baltimore, MD; 4 San Diego Digestive Disease Consultants, 

Inc., and Medical Associates Research Group, Inc., San Diego, 

CA; 5 Mayo Clinic, Pheonix, AZ; 6 eStudySite, San Diego, CA; 

7 University of Rochester Medical Center, Rochester, NY; 8 Virginia 

Mason Hospital and Medical Center, Seattle, WA; 9 Gastroenterology 

& Nutritional Medical Services, Bastrop, LA; 10 Texas Liver Institute, 

University of Texas Health Science Center, San Antonio, TX 

Background: The next-generation HCV direct acting antivirals 



characterized by potent pan-genotypic in vitro antiviral activity 

and a high barrier to resistance selection with retention of 

activity against key resistance-associated variants. ABT-493 

and ABT-530 each provided a mean 4 log 10 

IU/mL decline 

from baseline in HCV plasma viral load after 3 day monotherapy 

in genotype (GT) 1-infected subjects and have comparable 

in vitro antiviral potency against GT2. Purpose: To 

evaluate the efficacy and safety of ABT-493 and ABT-530 

with or without ribavirin (RBV) in non-cirrhotic GT2-infected 

treatment-naïve (TN) and pegylated interferon/RBV (pegIFN/ 

RBV) treatment experienced (TE) subjects Methods: Subjects 

received 12 weeks of ABT-493 300 mg+ABT-530 120 mg 

(Arm A), ABT-493 200 mg+ABT-530 120 mg (Arm B), or ABT- 

493 200 mg+ABT-530 120 mg+RBV (Arm C). DAAs were 

dosed once daily; weight-based RBV (1000 or 1200 mg) was 

dosed twice daily. Subjects were then followed for 24 weeks. 

The primary efficacy endpoint is sustained virologic response 

12 weeks after the last dose of study drug (SVR12); we present 

post-treatment week 4 data (SVR4) here. Safety was evaluated 

by monitoring adverse events (AEs), laboratory tests, 

and vital signs. Demographics and safety are presented on 

all randomized subjects. Efficacy is presented on all subjects 

confirmed to have GT2 infection. Results: 75 subjects were 

treated in Arms A–C (n=25 each); 74 had GT2, and 1 subject 

initially randomized to Arm B was determined to have GT3a 

infection. Subjects were male, 63%; median (range) age, 57.0 

(20.0–69.0) years; GT2b, 81%; TN, 88%; TE, 12%; F0–F2, 

87%; F3, 13%; median (range) baseline HCV RNA log 10 

IU/ 

mL, 7.1 (4.7–7.8). No subjects have experienced virologic 

failure. One subject in Arm A prematurely discontinued study 

drugs and is lost to follow-up. The SVR4 rates (ITT analysis) are 

96% (24/25), 100% (24/24), and 100% (25/25) in Arms


A, B, and C, respectively. Most AEs were mild, with the most 

common DAA-related AEs being fatigue, nausea, headache, 

and diarrhea. There were no serious DAA-related AEs. Typical 

reductions in hemoglobin were observed in the RBV-containing 

arm. Conclusions: ABT-493+ABT-530 with or without RBV for 

12 weeks was highly effective and well tolerated, achieving 

SVR4 rates of 96%–100% and no virologic failure to-date; 

SVR12 data will be available at the time of the meeting. These 

data warrant further study of this once daily RBV-free ABT-493/ 

ABT-530 combination in GT2 infection, including patients with 

cirrhosis. 

Disclosures: 






BMS 









Sandra S. Lovell - Employment: AbbVie 



AbbVie 



The following authors have nothing to disclose: Michael Bennett, Hugo E. Vargas, 

J. Scott Overcash, Asma Siddique, Bal R. Bhandari 

251 

High Efficacy of Grazoprevir and Elbasvir With or Without 

Ribavirin in 103 Treatment-Naive and Experienced 

Patients With HCV Genotype 4 Infection: A Pooled Analysis 

Tarik Asselah 1 , Hendrik W. Reesink 2 , Jan Gerstoft 3 , Victor de Ledinghen 

4 , Paul J. Pockros 5 , Michael Robertson 6 , Peggy Hwang 6 , 

Bach-Yen T. Nguyen 6 , Janice Wahl 6 , Eliav Barr 6 , Rohit Talwani 6 , 

Lawrence Serfaty 7 ; 1 Hôpital Beaujon, Clichy, France; 2 Academical 

Medical Center, Amsterdam, Netherlands; 3 Rigshospitalet, University 

of Copenhagen, Copenhagen, Denmark; 4 CHU Bordeaux, 

Pessac, France; 5 Scripps Clinic, La Jolla, CA; 6 Merck & Co., Inc., 

Kenilworth, NJ; 7 Saint-Antoine Hospital, Paris, France 

Background: Genotype (GT) 4 accounts for around 15% of the 

170 million HCV infections worldwide, with high prevalence 

in Middle East and sub-Saharan Africa, but also increasing in 

Europe. Data on the efficacy of direct acting antiviral regimens 

in the GT4 population are limited. We evaluated the efficacy 

of the protease inhibitor grazoprevir 100 mg (GZR) and NS5A 

inhibitor elbasvir 50 mg (EBR), with and without (±) ribavirin 

(RBV) in GT4 infected patients enrolled in the GZR/EBR 

phase 2/3 clinical program. Methods: Two pooled datasets 

were evaluated: (1) treatment-naive (TN) patients who received 

12 weeks of GZR/EBR ± RBV and (2) treatment-experienced 

(TE) patients, defined as those who had previously failed 

peginterferon + RBV (PR) ± first-generation protease inhibitor 

who received 12, 16, or 18 weeks of GZR/EBR ± RBV. TE 

patients were further classified as failures due to prior relapse 

or prior on-treatment virologic failure (OTF) (eg, null or partial 

response; virologic breakthrough). Efficacy was defined as sustained 

virologic response (HCV RNA < assay-specified lower 

limit of quantification) 12 weeks after end of therapy (SVR12) 

in the full analysis set (all patients who received at least 1 dose 

of study drug). Results: Data from a total of 103 HCV-GT4 (47 

GT4a, 47 GT4d, 9 GT4-other)–infected patients, 66 TN and 

37 TE, including 17 cirrhotic patients, were analyzed. Overall 

64/66 (97%) TN (including 6/6 cirrhotic patients) and 32/37 

(86%) TE GT4 patients achieved SVR12. Among TE patients, 

9/9 prior relapsers and 23/28 (82%) prior OTFs achieved 

SVR12. Among TE cirrhotic patients, 13/17 (77%) including 

3/3 prior relapsers and 4/4 treated with GZR/EBR+RBV for 

16 weeks achieved SVR12. For prior OTFs, longer durations 

and use of RBV increased the response rate. Additional stratification 

of treatment responses according to regimen is included 

in Table 1. Conclusion: A 12-week regimen of GZR/EBR was 

highly effective among 103 GT4-infected TN and prior relapse 

patients. A regimen of GZR/EBR + RBV for 16 weeks may 

result in improved efficacy among prior OTF patients, although 

sample size was small. Further studies among subpopulations 

are needed. 

Table 1 

a Other: Non-SVR, but did not meet virologic failure criteria 

Disclosures: 



Hendrik W. Reesink - Consulting: Abbvie, Alnylam, BMS, Gilead, Janssen-Cilag, 

Merck, PRA-International, Regulus, Replicor, Roche, R-Pharm; Grant/Research 

Support: AbbVie, BMS, Boehringer Ingelheim, Gilead, Janssen-Cilag, Merck, 

PRA-International, Regulus, Replicor, Roche 








Peggy Hwang - Employment: Merck, Merck 




Rohit Talwani - Employment: Merck 

Lawrence Serfaty - Advisory Committees or Review Panels: MSD, Janssen, Roche, 

Gilead, BMS, Abbvie; Speaking and Teaching: Aptalis 

The following authors have nothing to disclose: Jan Gerstoft


252 

Daclatasvir in combination with sofosbuvir with or without 

ribavirin is safe and efficacious in liver transplant 

recipients with HCV recurrence: Interim results of a multicenter 

compassionate use program 

Kerstin Herzer 1 , Tania M. Welzel 2 , Peter Ferenci 3 , Joerg Petersen 4 , 

Michael Gschwantler 5 , Markus Cornberg 6 , Thomas Berg 7 , Ulrich 

Spengler 8 , Ola Weiland 9 , Marc van der Valk 10 , Hartwig Klinker 11 , 

Jürgen K. Rockstroh 8 , Eckart Schott 12 , Markus Peck-Radosavljevic 3 , 

Yue Zhao 13 , Maria Jesus Jimenez Exposito 14 , Stefan Zeuzem 2 ; 

1 Universitätsklinikum Essen (AöR), Essen, Germany; 2 Universitätsklinikum 

der Johann Wolfgang Goethe Universität, Frankfurt, Germany; 

3 Medizinische Universität Wien, Vienna, Austria; 4 IFI Institut 

für Interdisziplinäre Medizin, Hamburg, Germany; 5 Wilhelminenspital, 

Vienna, Austria; 6 Medizinische Hochschule Hannover, Hannover, 

Germany; 7 Universitätsklinikum Leipzig, Leipzig, Germany; 

8 Universitätsklinikum Bonn, Bonn, Germany; 9 Karolinska Institutet, 

Solna, Sweden; 10 Academic Medical Center, University of Amsterdam,, 

Amsterdam, Netherlands; 11 Universitätsklinikum Würzburg, 

Würzburg, Germany; 12 Charité Universitätmedizin Berlin, Berlin, 

Germany; 13 Global Biostastistics, Bristol-Myers Squibb, Hopewell, 

NJ; 14 Research and Development, Bristol-Myers Squibb, Princeton, 

NJ 

BACKGROUND: Interferon (IFN)-based therapy for HCV postliver 

transplant (LT) has been associated with suboptimal 

response and poor tolerability. New IFN-free regimens have 

the potential to improve response rates and safety. The all-oral, 

pangenotypic regimen of daclatasvir (DCV) and sofosbuvir 

(SOF) with ribavirin (RBV) was well tolerated and achieved 

high SVR rates in patients with post-LT recurrence or advanced 

cirrhosis in the ALLY-1 study. We report here a detailed subanalysis 

of LT recipients with HCV recurrence from a compassionate 

use program (CUP) of DCV+SOF±RBV. METHODS: This 

multicenter CUP, opened in 5 European countries, enrolled 

adult patients with chronic HCV infection who are at a high 

risk of hepatic decompensation or death within 12 months if 

left untreated, and who had no available treatment options. 

Patients received DCV 60mg + SOF 400mg once daily for 

24 weeks, with RBV added at the physician’s discretion. This 

interim analysis includes 87 post-LT patients enrolled in the CUP 

with available data on March 16, 2015, of whom 38 patients 

have SVR12 data available to date. RESULTS: Most patients 

were male (69%), white (93%), with a median age of 58 years 

(39–75). Mean (SD) baseline HCV RNA 5.8 (1.7) log 10 

IU/ 

mL. Most patients were infected with HCV GT1 (87%) (1a, 

n=28; 1b, n=37; 1 unknown, n=11) or GT3 (8%). Median 

time since LT to treatment initiation was 3.3 years. Most frequent 

immunosuppressant includes tacrolimus (n=47) and cyclosporine 

(n=11). Cirrhosis was present in 35 (40%) patients; 

among cirrhotic patients, 13 (37%) were Child-Pugh class B/C, 

and 8 (23%) had MELD scores ≥15. Twenty-six (30%) patients 

received RBV as part of the regimen. Baseline characteristics 

and results for each treatment group are described in the Table. 

Overall, of the 38 patients with available HCV RNA values at 

posttreatment week 12 to date, all achieved virologic response 

(SVR12 100%). Updated SVR12 data will be presented. Serious 

AEs occurred in 18 patients (21%), 6 patients discontinued 

treatment due to AEs and 3 patients died. CONCLUSION: In 

this preliminary analysis of patients in a real-world setting, 

DCV+SOF±RBV achieved high rates of SVR12 and was well 

tolerated in this difficult-to-treat post-liver transplant patient population. 

The use of ribavirin did not affect efficacy outcomes. 

Disclosures: 































Eckart Schott - Advisory Committees or Review Panels: Gilead, Roche, Bayer, 

BMS, Abbvie; Speaking and Teaching: Gilead, Novartis, Roche, MSD, Bayer, 

Falk, BMS, Janssen, Abbvie 








The following authors have nothing to disclose: Kerstin Herzer, Ulrich Spengler, 

Hartwig Klinker, Yue Zhao 

253 

Molecular heterogeneity in multinodular Hepatocellular 

Carcinoma 

Daniela Sia 1,2 , Andrew Harrington 1 , Zhongyang Zhang 3 , Genis 

Camprecios 1 , Sara Toffanin 1 , Agrin Moeini 1,2 , M. Isabel Fiel 1 , 

Ke Hao 3 , Monica Higuera 2 , Oriana Miltiadous 1 , Laia Cabellos 2 , 

Helena Cornella 2 , Yujin Hoshida 1 , Sander S. Florman 1 , Myron E. 

Schwartz 1 , Josep M. Llovet 1,2 ; 1 Liver Cancer Program, Dept of Liver 

Diseases, Icahn School of Medicine at Mount Sinai, New York, 

NY; 2 Liver Cancer Translational Research Laboratory, IDIBAPS, 

Hospital Clinic, Barcelona, Spain; 3 Icahn Institute for Multiscale 

Biology, Icahn School of Medicine at Mount Sinai, New York, NY 

Introduction: Whether multinodular hepatocellular carcinomas 

(HCCs) result from intrahepatic metastasis (IM or clonal tumors)


or de novo cancers (synchronic tumors) has direct implications 

in treatment decision-making. We aimed to assess the genomic 

heterogeneity of multifocal HCC using single-nucleotide polymorphism 

(SNP), gene expression and mutations analyses. 

Methods: Among 544 HCCs consecutively transplanted at 

Mount Sinai Hospital, 18 patients with 2-3 non-satellite nodules/patient 

were selected for this study. Formalin-fixed tissue 

blocks and clinico-pathological parameters were collected for 

each tumor. SNP array and gene expression profiling was generated. 

Clonality was defined by measuring SNP profiles similarity 

between two nodules. Prediction of previously published 

signatures was performed using Nearest Template Prediction. 

Mutational status of TERT promoter, ARID1A, CTNNB1, TP53, 

AXIN1-2 genes was checked using TruSeqAmplicon. Results: 

A total of 42 tumors have been analyzed. Most patients were 

male (17/18, 95%) with HCV (10/18, 56%) or HBV infection 

(6/18, 33%). Median tumor size was 3 cm (range 1.5- 

6.5), satellites and vascular invasion were present in 3 (17%) 

and 11 patients (61%), respectively. CNV profiles predicted 

clonal tumors in 38% (6/16) and non-clonality in 63% of cases 

(10/16), while the remaining 2 cases were not informative. 

Clonal tumors were significantly associated with HCV infection 

(5/6 vs 3/10, p=0.01), whereas all HBV-induced HCC were 

synchronic tumors (0/6 vs 6/10, p=0.03). Clonal tumors were 

significantly associated with presence of satellites and early 

recurrence. Gene expression-based unsupervised clustering 

revealed that each clonal tumor showed genetic proximity to its 

paired tumor and clustered around the same node (6/6,100%) 

as opposed to non-clonal (2/9, 22%). When exploring molecular 

subclasses, while half of clonal tumors retained the molecular 

fingerprint, the other half switched to more aggressive 

subclasses. Conversely, all non-clonal tumors belonged to distinct 

molecular subclasses. In order to explore if clonal tumors 

retain the same driver trunk mutations, we tested five common 

driver mutations in all HCC nodules. Identical trunk mutations 

were identified only in clonal tumors. Conclusions: Among 

multinodular HCC tumors, 40% of cases were classified as 

clonal tumors (true IM) and the remaining 60% as non-clonal 

synchronic tumors. Clonal tumors were significantly associated 

with HCV infection, satellites and recurrence. Despite molecular 

subclasses prediction revealed heterogeneous profile in 

both clonal and non-clonal tumors, genetic proximity and trunk 

driver mutations were similar in clonal tumors. 

Disclosures: 

Josep M. Llovet - Consulting: Bayer Pharmaceuticals, Bristol Myer Squibb, Boehringer-Ingelheim, 

Eli Lilly Pharmaceuticals, Celsion, Biocompatibles, Novartis, 

GlaxoSmithKline, Blueprint Medicines; Grant/Research Support: Bayer Pharmaceuticals, 

Bristol Myers Squibb, Boehringer-Ingelheim 

The following authors have nothing to disclose: Daniela Sia, Andrew Harrington, 

Zhongyang Zhang, Genis Camprecios, Sara Toffanin, Agrin Moeini, M. Isabel 

Fiel, Ke Hao, Monica Higuera, Oriana Miltiadous, Laia Cabellos, Helena Cornella, 

Yujin Hoshida, Sander S. Florman, Myron E. Schwartz 

254 

Mixed hepatocellular-cholangiocarcinoma tumors: 

stem-cell subtype and cholangiolocellular carcinoma are 

unique molecular entities 

Agrin Moeini 1,2 , Daniela Sia 2,1 , Zhongyang Zhang 3 , Genis 

Camprecios 2 , M. Isabel Fiel 2 , Oriana Miltiadous 2 , Xiaochen 

Sun 2 , Ke Hao 3 , Yujin Hoshida 2 , Swan N. Thung 2 , Augusto Villanueva 

2 , Myron E. Schwartz 2 , Josep M. Llovet 2,1 ; 1 Liver Cancer 

Translational Research Laboratory, Liver Unit, Institut d’Investigacions 

Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clínic, 

CIBERehd, Universitat de Barcelona, Barcelona, Spain; 2 Mount 

Sinai Liver Cancer Program, (Divisions of Liver Diseases, Department 

of Medicine, Department of Pathology, Recanati Miller 

Transplantation Institute), Tisch Cancer Institute, Icahn School 

of Medicine at Mount Sinai, New York, NY; 3 Icahn Institute for 

Genomics and Multiscale Biology, Icahn School of Medicine at 

Mount Sinai, New York, NY 

Purpose: Provide a comprehensive analysis of the molecular 

alterations of mixed hepatocellular-cholangiocarcinoma (HCCiCCA) 

tumors, a rare type of primary liver cancer, by integrating 

histological and genomic data. Methods: 19 patients 

with mixed HCC-iCCA were classified based on histological 

features and immunohistochemistry for specific hepatocyte 

(HEP1, GPC3), biliary (CK7, CK19) and stem-cell markers 

(EpCAM, NCAM, SALL4). Whole-genome gene-expression 

and DNA copy number alterations (CNA) analysis were performed 

(Illumina). Prevalent oncogenic mutations in HCC (TERT 

promoter, TP53, CTNNB1) or iCCA (FGFR2-fusions, IDH1/2, 

KRAS, BRAF) were screened. Results: Following WHO criteria, 

HCC-iCCA samples were classified as classical (n=4), stem-cell 

feature (n=9) and cholangiolocellular carcinoma (CLC) (n=6). 

From the pathological standpoint, classical HCC-iCCA were 

mainly EpCAM positive (3/4, 75%) with expression of hepatocyte 

and biliary markers in the HCC-like and iCCA-like components, 

respectively. Stem-cell subtypes were characterized 

by SALL4 positive staining (7/9 vs 0/10 others, p


Disclosures: 

Josep M. Llovet - Consulting: Bayer Pharmaceuticals, Bristol Myer Squibb, Boehringer-Ingelheim, 

Eli Lilly Pharmaceuticals, Celsion, Biocompatibles, Novartis, 

GlaxoSmithKline, Blueprint Medicines; Grant/Research Support: Bayer Pharmaceuticals, 

Bristol Myers Squibb, Boehringer-Ingelheim 

The following authors have nothing to disclose: Agrin Moeini, Daniela Sia, 

Zhongyang Zhang, Genis Camprecios, M. Isabel Fiel, Oriana Miltiadous, Xiaochen 

Sun, Ke Hao, Yujin Hoshida, Swan N. Thung, Augusto Villanueva, Myron 

E. Schwartz 

255 

Loss of adherens junctions proteins disrupts blood 

bile barrier to cause progressive familial intrahepatic 

cholestasis (PFIC)-like phenotype 

Kari Nejak-Bowen, Lili Zhou, An Jiang, Minakshi Poddar, Satdarshan 

(Paul) S. Monga; University of Pittsburgh, Pittsburgh, PA 

Tight junction (TJ) proteins are traditionally thought to be the 

chief regulators of blood-bile barrier in the liver. In fact, loss 

of TJP2 has been shown to be associated with a subset of 

PFIC cases. PFIC is traditionally linked to loss of function mutations 

in various bile acid transporters such as BSEP, MDR3 and 

ATP8B1, although the molecular basis of a notable subset of 

these cases remains elusive. Further, the role of adherens junctions 

(AJ) in regulating blood-bile barrier or in PFIC pathogenesis 

remains unknown. AJ regulate cell-cell adhesion by linking 

E-cadherin of neighboring cells to respective actin cytoskeleton 

through β-catenin and α-catenin. Intriguingly, we showed that 

conditional loss of β-catenin in hepatocytes was well compensated 

by upregulation of γ-catenin, a desmosomal protein that 

maintained the E-cadherin-actin cytoskeleton link. To characterize 

this redundancy in catenin proteins, we generated 

liver-specific β-catenin-γ-catenin double knockout (DKO) mice 

using cre-lox. DKO show failure to thrive and most animals 

die by 1 month. Kaplan Meier curve shows significantly lower 

survival as compared to littermate controls (Con) (p


257 

Hepatic XBP1 deletion promotes endoplasmic reticulum 

stress-induced liver injury and apoptosis 

Shantel Olivares, Anne Henkel; Medicine, Northwestern University, 

Chicago, IL 

Endoplasmic reticulum (ER) stress triggers activation of the 

unfolded protein response (UPR), a highly conserved signaling 

cascade that functions to alleviate stress and promote cell survival. 

If, however, the cell is unable to restore homeostasis, the 

UPR activates pathways that promote apoptotic cell death. The 

molecular mechanisms governing this critical transition from restoration 

of homeostasis to initiation of apoptosis remain poorly 

understood. We aim to determine the role of hepatic XBP1, a 

key regulator of the UPR, in controlling cell fate in response 

to ER stress. Liver-specific XBP1 knockout mice (XBP1 LKO ) and 

XBP1 fl/fl littermate controls were subjected to varying levels and 

duration of pharmacologic ER stress. XBP1 LKO mice and XBP1 fl/ 

fl 

controls showed robust and equal induction of the UPR acutely 

as evidenced by equivalent hepatic expression of Grp78/BIP, 

CHOP, and ATF4 six hours after exposure to pharmacologic 

ER stress. By 72 hours, XBP1 fl/fl control mice showed complete 

resolution of UPR activation and no liver injury suggesting complete 

restoration of homeostasis. Conversely, XBP1 LKO mice 

showed ongoing UPR activation at 3 and 7 days after induction 

of ER stress associated with progressive liver injury histologically. 

Consistent with enhanced hepatic injury, plasma ALT 

levels were markedly elevated in XBP1 LKO mice at days 3 and 7 

(283±41 vs 49±12 IU/L at day 3 and 217±3 vs 47±11 IU/L at 

day 7 vs XBP1 fl/fl controls, p


259 

Midodrine and Tolvaptan in Patients with Cirrhosis and 

Refractory or Recurrent Ascites : A Randomized Pilot 

Study 

Virendra Singh 1 , Nitish Rai 2 , Baljinder Singh 3 , Rajesh Vijayvergiya 

4 , Navneet Sharma 2 , Ashish Bhalla 2 ; 1 Hepatology, 

Postgraduate Institute of Medical Education and Research, Chandigarh, 

India; 2 Internal Medicine, Postgraduate Institute of Medical 

Education and Research, Chandigarh, India; 3 Nuclear Medicine, 

Postgraduate Institute of Medical Education and Research, Chandigarh, 

India; 4 Cardiology, Postgraduate Institute of Medical Education 

and Research, Chandigarh, India 

Background: Splanchnic arterial vasodilatation and subsequent 

activation of antinatriuretic and vasoconstrictive mechanisms 

resulting in sodium and water retention play an important role 

in cirrhotic ascites. Midodrine and tolvaptan have been used 

in these patients, separately. However, there are no reports 

on the use of combination of midodrine and tolvaptan in the 

control of ascites. The aim of this study was to evaluate the 

effects of midodrine,tolvaptan and their combination on systemic 

hemodynamics, renal function and control of ascites in 

patients with cirrhosis and refractory or recurrent ascites. Methods: 

Fifty cirrhotic patients with refractory or recurrent ascites 

were prospectively studied after long-term administration of 

midodrine (7.5 mg three times a day; n=13) or tolvaptan (15 

mg twice a day; n=12) or both (n=13) plus standard medical 

therapy (SMT) or SMT alone (n=12) in a randomized controlled 

trial (NCT02173288) at a tertiary center. Results: A 

significant increase in urinary volume (midodrine:1077±274 

vs 1440±270 ml/day, p=0.004; tolvaptan:1150±344 vs 

2250±688 ml/day, p=0.001; combination:1130±233 

vs 3110±1170 ml/day, p=0.0001 at 1 and 3 months, 

p


Peter C. Hayes - Advisory Committees or Review Panels: Roche, MSD, Jannsen, 

Gilead, ?ONO, Norgine; Grant/Research Support: Novartis; Speaking and 

Teaching: Roche, MSD, Pfiser, Gore, Falk, Ferring 

The following authors have nothing to disclose: Neil J. Lachlan, Scott I. Semple, 

Dilip Patel, David Carr, John P. Iredale, Jonathan A. Fallowfield 

261 

Ascites neutrophil dysfunction in patients with decompensated 

cirrhosis is linked to ascites protein content 

Cornelius Engelmann 1 , Christina Becker 2 , Andreas Boldt 2 , Sandra 

Krohn 1 , Michael Bartels 3 , Thomas Berg 1 , Ulrich Sack 2 ; 1 Section 

of Hepatology; Department of Internal Medicine, Neurology, Dermatology, 

University Hospital Leipzig, Leipzig, Germany; 2 Institute 

of Clinical Immunology, University of Leipzig, Leipzig, Germany; 

3 Department of Visceral, Vascular, Thoracic and Transplant Surgery, 

University Hospital Leipzig, Leipzig, Germany 

Background: Spontaneous bacterial peritonitis is a major and 

life threatening complication in patients with decompensated 

cirrhosis. Immune paralysis and neutrophil dysfuntion has been 

regarded as a main driver for the increased risk of bacterial 

infections in these patients. It remains unknown whether this 

feature can be extrapolated to immune cells in the peritoneal 

cavity. We therefore evaluated the cell function of neutrophils 

(PMN) in ascites fluid. Material and Methods: In total 71 blood 

and ascites samples of mainly male patients (n=48) with cirrhosis 

(n=63) and without cirrhosis (malignant n=6, cardiac n=1, 

Budd-Chiari syndrome n=1) were collected between August 

2014 and May 2015. PMNs were identified by flow-cytometry. 

Phagocytic activity (PA) as well as oxidative burst activity 

(OBA) of both blood and ascites PMNs were evaluated after 

in vitro stimulation with E.coli cells. Fluorescence signal was 

measured by flow cytometry and PA and OBA was expressed 

as percentage of viable PMNs. Function tests of ascites PMNs 

were repeated in human albumin and autologous blood 

plasma. Results: As compared to their blood counterparts, ascites 

PMNs showed a significantly impaired PA and OBA with a 

much broader activity range (median blood PA (n=70) 98.1% 

(86.8-99.8) vs. median ascites PA (n=68) 52.95% (0.4-97.3), 

p


263 

TIPS with PTFE-covered stent improves liver transplant 

free survival in patients with cirrhosis and recurrent 

ascites : results of a multicentre randomized trial 

Christophe Bureau 1,2 , Dominique Thabut 3 , Frederic Oberti 4 , 

Sebastien Dharancy 5 , Pauline Cabarrou 1 , Nicolas Carbonell 3 , 

Antoine Bouvier 4 , Philippe Mathurin 5 , Philippe Otal 1 , Jean-Marie 

Peron 1,2 , Jean-Pierre Vinel 1,2 ; 1 Service d’Hépatogastroenterologie, 

CHU Toulouse, Toulouse, France; 2 Université Paul Sabatier, 

Toulouse, France; 3 APHP, Paris, France; 4 CHU Angers, Angers, 

France; 5 CHRU Lille, Lille, France 

Introduction Transjugular intrahepatic portosystemic shunt 

(TIPS) proved to be effective in the treatment of refractory ascites. 

However, its impact on survival remains controversial. The 

high rate of shunt dysfunction with the use of uncovered stents 

could attenuate the beneficial effects of TIPS. It has been shown 

that the use of PTFE covered stents decreases the risk of shunt 

dysfunction and improves clinical outcome. The main objective 

was to compare the liver transplantation free survival of 

cirrhotic patients and recurrent ascites between those treated 

by covered TIPS and those treated by large volume paracentesis 

+ albumin infusion (P+A). Methods Between august 2005 

and november 2012, all cirrhotic patients with at least two 

large volume paracentesis were considered for inclusion. The 

main end point was 1-year liver transplantation free survival. 

Secondary end points were the rates of: ascites recurrence 

and treatment failure, overt hepatic encephalopathy and portal 

hypertension (PHT) related complications; and the number of 

days in hospitalization over a 1-year period after inclusion. 

Results 62 patients were included (29 in TIPS group and 33 in 

P+A group). The baseline characteristics of the patients were 

similar in both groups. The median follow up of the whole 

cohort was 365 days. One patient in each group was lost to 

follow up. Actuarial rates of transplantation free survival was 

better in TIPS group compared to that observed in the P+A 

group: 93 % [IC 95 % 82%-100 %] and 52 % [IC 95 % 34%- 

60 %], respectively (p=0.003). During the 1-year of follow 

up, the total number of paracentesis was 32 (1.0 ± 1.6 per 

patient) in TIPS group and 320 in P+A (10.1 ± 7.0 per patient). 

Total albumin infusion was 2061 g in TIPS group compared to 

17727 g in P+A group. In P+A group, 15 patients were treated 

by TIPS during follow up, 14 because of the need of more than 

6 large volume paracentesis within less than 3 months and 1 

because of associated recurrent variceal bleeding. The 1-year 

probability of remaining free of encephalopathy was 65 % 

in both groups. The rates of PHT related bleeding and hernia 

related complications were significantly higher in P+A group. 

The number of days in hospitalizations was doubled in P+A 

group compared to TIPS group (17 ± 28 vs 35 ± 40 p = 0.04). 

Conclusion: the use of covered TIPS in selected patients with 

recurrent ascites improved transplant free survival as compared 

to patients treated by repeated large volume paracentesis + 

albumin infusion. 

Disclosures: 

Christophe Bureau - Grant/Research Support: Gore; Speaking and Teaching: 

Gore 

Frederic Oberti - Stock Shareholder: Bio Live Scale 

Sebastien Dharancy - Advisory Committees or Review Panels: CHIESI; Board 

Membership: NOVARTIS; Speaking and Teaching: ASTELLAS 

Philippe Mathurin - Board Membership: MSD, Janssen-Cilag, BMS, Gilead, 

Abvie, Oncozyme; Consulting: Roche, Bayer 

Jean-Marie Peron - Board Membership: BAYER; Consulting: BMS, GILEAD, BOS- 

TON SCIENTIFIC 

Jean-Pierre Vinel - Grant/Research Support: Roche, Gore, LFB 

The following authors have nothing to disclose: Dominique Thabut, Pauline 

Cabarrou, Nicolas Carbonell, Antoine Bouvier, Philippe Otal 

264 

Diagnostic accuracy of the Periscreen TM reagent strip 

in diagnosis of spontaneous bacterial peritonitis in cirrhotic 

patients (PerDRISLA study) 

Thierry Thevenot 1 , Charline Briot 1 , Vincent Macé 2,22 , Hortensia 

Lison 3 , Laure Elkrief 4 , Alexandra Heurgué-Berlot 5 , Christophe 

Bureau 6 , Caroline Jezequel 7 , Ghassan Riachi 8 , Alexandre Louvet 

9 , Isabelle Ollivier-Hourmand 10 , Hélène Labadie 11 , Nicolas 

Carbonell 12 , Karim Aziz 13 , Denis Grasset 14 , Rodolphe Anty 15 , 

Eric Nguyen-Khac 16 , Mehdi Kaasis 17 , Thomas Mouillot 18 , Arnaud 

Pauwels 19 , Florence Tanné 20 , Si Nafa Si Ahmed 21 , Jean Paul 

Cervoni 1 , Jean françois D. Cadranel 3 , Matthieu Schnee 2 ; 1 Hepatogastroenterology, 

CHU Minjoz, Besançon, France; 2 Hepatogastroenterology, 

CHD-Vendée, La Roche sur Yon, France; 

3 Hepatogastroenterology, Höpital Laennec, Creil, France; 4 Hepatology, 

CHU- Beaujon, Clichy, France; 5 Hepatogastroenterology, 

CHU Reims, Reims, France; 6 Hepatogastroenterology, CHU Purpan, 

Toulouse, France; 7 Hepatogastroenterology, CHU Pontchaillou, 

Rennes, France; 8 Hepatogastroenterology, CHU Charles 

Nicolle, Rouen, France; 9 Hepatology, CHU Claude Huriez, Lille, 

France; 10 Hepatogastroenterology, CHU Caen, Caen, France; 

11 Hepatogastroenterology, CH de Saint-Denis, Saint-Denis, 

France; 12 Hepatogastroenterology, Hôpital Saint-Antoine, Paris, 

France; 13 Hepatogastroenterology, CH de Saint-Brieuc, Saint- 

Brieuc, France; 14 Hepatogastroenterology, CH Bretagne-Atlantique, 

Vannes, France; 15 Hepatogastroenterology, CHU Archet, 

Nice, France; 16 Hepatogastroenterology, CHU Amiens, Amiens, 

France; 17 Hepatogastroenterology, CH de Cholet, Cholet, France; 

18 Hepatogastroenterology, Hôpital du Bocage, Dijon, France; 

19 Hepatogastroenterology, CH de Gonesse, Gonesse, France; 

20 Hepatogastroenterology, CHU La-Cavale-Blanche, Brest, France; 

21 Hepatogastroenterology, CHR d’Orléans, Orléans, France; 

22 Hepatogastroenterology, CHU de Nantes, Nantes, France 

Background & Aim: The diagnostic accuracy of spontaneous 

bacterial peritonitis (SBP) using reagent strips is currently disappointing 

(1) but an experimental work using the Periscreen 

TM strip was more optimistic (2). We aim to assess the 

performance of the Periscreen TM strip for the diagnosis of SBP. 

Methods: This study involved all consecutive cirrhotic patients 

with ascites between June 2014 and March 2015 among 

22 French centers. Ascitic fluid was tested independently by 

two investigators using Periscreen TM at 3 minutes as reported 

(2). The strip has 4 colorimetric graduations (negative, trace, 

small or large). The diagnosis of SBP was based on neutrophils 

>250/mm 3 , regardless the culture of ascites. We excluded 

bloody, chylous or bilious ascitic fluid, or concurrent imipenem 

treatment. Results: The characteristics of the 387 cirrhotic 

patients included were: mean age 61.9±10.4 years, 75.4% 

males, 73.1% alcohol, mean Child-Pugh 9.6 (range: 6-15). A 

total of 1190 paracenteses were analyzed: 468 (39.3%) were 

collected in patients hospitalized for an acute decompensation 

of cirrhosis and 722 (60.7%) in outpatients. For inter-investigator 

agreement, the Kappa value was 0.79 (IC95%: 0.76- 

0.83). A total of 72 (5.8%) samples had SBP, 58 in inpatient 

(11.5%) and 14 (1.9%) in outpatient setting (P


(64.9-83.4), PPV 34.3% (19.1-52.2) and VPN 97.2% (90.2- 

99.7). In this study, the prevalence of SIRS was 13.2%. In 

5 samples, neutrophil count was above 250/mm 3 (385 to 

13020/ mm 3 ) but the strip was considered as negative. When 

setting “trace” as the first positive result, Se and NPV were both 

of 100% for outpatients. Conclusions: This study shows, for the 

first time, in a large sample size of cirrhotic patients that the 

Periscreen TM strip is a robust screening tool for the detection of 

SBP. More importantly, this new leukocyte esterase strip has 

an excellent accuracy to exclude SBP in the outpatient setting. 

(1) Nousbaum JB, et al. Hepatology 2007;45:1275-1281. (2) 

Mendler MH, J Hepatol 2010;53:477-483. 

Disclosures: 

Alexandra Heurgué-Berlot - Consulting: Abbvie; Speaking and Teaching: Gilead 


Gore 

Isabelle Ollivier-Hourmand - Speaking and Teaching: gilead, jansen, bayer 


MSD, BMS 

Si Nafa Si Ahmed - Board Membership: La Roche Hoffmann; Grant/Research 

Support: Janssen; Speaking and Teaching: BMS, Gilead, Abbvie 

The following authors have nothing to disclose: Thierry Thevenot, Charline Briot, 

Vincent Macé, Hortensia Lison, Laure Elkrief, Caroline Jezequel, Ghassan Riachi, 

Alexandre Louvet, Hélène Labadie, Nicolas Carbonell, Karim Aziz, Denis Grasset, 

Rodolphe Anty, Mehdi Kaasis, Thomas Mouillot, Arnaud Pauwels, Florence 

Tanné, Jean Paul Cervoni, Jean françois D. Cadranel, Matthieu Schnee 

265 

Deleterious effect of beta-blockers in cirrhotic patients 

with refractory ascites: potential role of myocardial dysfunction 

Valerio Giannelli 1,2 , Olivier Roux 2 , Pierre-Emmanuel Rautou 2 , 

Emmanuel Weiss 2 , Richard Moreau 2 , Didier Lebrec 2 , Francois 

Durand 2 , claire francoz 2 ; 1 gastroenterology, university of rome 

“sapienza”, Rome, Italy; 2 Service d’Hépatologie, Hôpital Beaujon, 

Assistance Publique-Hôpitaux de Paris, Clichy, France; INSERM 

U1149,CRI, Paris, France 

Introduction: Recent studies have suggested that non selective 

beta-blockers (NSBB) could be deleterious in cirrhotic patients 

with refractory ascites (RA) by increasing transplant-free mortality. 

It has been proposed that NSBB may increase the risk 

of paracentesis-induced circulatory dysfunction but the mechanisms 

involved are still unclear. The aim of this study was to 

explore the impact of NSBB on systemic and splanchnic hemodynamics 

in cirrhotic patients with RA evaluated for liver transplantation 

(LT). Patients and methods: 583 cirrhotic patients 

evaluated for a first LT in a single center between 1997 and 

2013 were studied. There were 74% males, mean age was 

52 years. The cause of cirrhosis was alcohol in 43%, HCV 

in 24% and HBV in 11%. 29% of patients had hepatocellular 

carcinoma. 196 patients (34%) had RA, and among them, 51 

% received NSBB. Physiological MELD score was comparable 

in RA and no—RA groups (17 vs 16, ns) as well as the proportion 

of patients receiving NSBB (51 vs 51 %, ns). In addition 

to standard preLT workup, all patients had measurement of 

both splanchnic and systemic hemodynamics by right heart 

catheterization with assessment of cardiac work (Left and Right 

Ventricular Stroke Work Index, LVSWI and RVSW). Results: 

Waiting list mortality was higher in RA compared to no-RA 

group, (62 vs 38 % at 1 year, p=0.001). RA patients had significantly 

lower mean arterial pressure (MAP) lower heart rate 

(HR) and higher hepatic venous pressure gradient (HVPG) than 

no-RA patients. Cardiac index (CI) as well as systemic vascular 

resistance (SVRI) were similarcomparable in both groups. By 

contrast, Left and Right Ventricular Stroke Work Index (LVSWI 

and RVSWI) were significantly lower in RA patients compared 

to no-RA. In both RA and no-RA patients, NSBB resulted in a 

significant decrease in HR, MAP, and CI. In the no-RA group, 

NSBB did not affect LVSWI (57+15 vs 60+21 g.m/m 2 , ns). By 

contrast, in the RA group, NSBB were associated with a significant 

decrease in LVSWI (48+15 vs 56+15, p


48%, and 33%, respectively. In multivariate analysis, AKI 

stage1B or greater (HR=2.32; p


269 

Hepatiq Measure Of Hepatic Function: Threshold Function 

For Ascites And Death In Patients With Chronic Liver 

Disease (CLD) 

John C. Hoefs 1,2 , Lien Tran 2 , Dipu Ghosh 3 ; 1 Medicine, Univ of 

California, Orange, CA; 2 Medicine, Liver Specialtly Center, Irvine, 

CA; 3 HEPATIQ_LLC, Irvine, CA 

INTRODUCTION:Functional quantitative tests have been 

largely ignored in the non-invasive staging of CLD. The perfused 

hepatic mass (PHM) is a precise measure of liver function 

correlating with the functional hepatic mass (r2 = .905)(AmJ- 

Gastro;92:2054) and adverse clinical outcomes (ACO). PHM 

= 95 (HALT-C 

2012;Hepat;55:1019). HEPATIQ is an automated measure 

of PHM to make quantitative image analysis of SPECT images 

easier. METHODS: Sequential SPECT scans from 384 patients 

were processed by HEPATIQ for PHM to assess the relationship 

to ascites and death. There were normals (N) 29 and patients 

with CLD: HBV 61, HCV 138, NASH 49, PBC/ACAH/PSC 

34, ALD 13, abnormal AST/ALT44, post liver transplant (LT) 

6, and miscellaneous 16. Any ascites that required treatment 

was recorded. Patients with recovery from prior ACO from 

CLD associated with ascites (R-ACO: requiring no treatment > 

2 yrs) or before liver transplant (LT) and prior variceal bleeding 

without ascites (VB) were included along with treatable ascites 

(RxA), Refractory ascites (RfA), and death (D). SCAN: Patients 

were fed prior to IV injection of 5-6 mCi 99Tc sulfur colloid with 

subsequent SPECT /planar images and processed by HEPA- 

TIQ. RESULTS: 306 patients never had ACO, 5 had a variceal 

bleed with no ascites (VB) and 28 patients had current ascites: 

responsive ascites RxA 10, refractory ascites (RfA) 12, and 

death 6. PHM in N was 104.3+/-2.8 and 306 patients never 

having ACO 102.3+/-4.6. In 6 LT patients PHM 99.2+/-4.5 

and R-ACO 91.9+/-6.4. PHM was 77.7+/-6.9 with RxA; RfA 

64.2+/-11.6 and 6 deaths 49.9+/-16.1 (< 60 for 4 liver failure, 

2 HCC). For PHM threshold for ACO PHM < 95, RxA 85, 

RfA 78, and death < 60 (see figure) CONCLUSIONS: 1. PHM 

using HEPATIQ is a precise measure of CLD severity correlating 

with clinical outcomes regardless of CLD cause. 2. Hepatic 

function by PHM thresholds for RxA, RfA, and death are relatively 

precise, 3. PHM with HEPATIQ is a hepatic function test, 

valuable for non-invasive staging and monitoring of CLD. 

Disclosures: 

John C. Hoefs - Management Position: HEPATIQ_LLC; Speaking and Teaching: 

Gilead, Jannsen, Abbvie 

Dipu Ghosh - Management Position: HEPATIQ LLC 

The following authors have nothing to disclose: Lien Tran 

270 

The Trigger Matters – Outcome of Specifically-Triggered 

AKI versus HRS in Patients with Cirrhosis and Ascites 

Theresa Bucsics, Philipp Schwabl, Mattias Mandorfer, Simona 

Bota, Bernhard Scheiner, Wolfgang Sieghart, Arnulf Ferlitsch, 

Michael Trauner, Markus Peck-Radosavljevic, Thomas Reiberger; 

Div. of Gastroenterology and Hepatology, Department of Internal 

Medicine III, Medical University of Vienna, Vienna, Austria 

Background: Hepatorenal syndrome (HRS) is considered a 

severe and often lethal type of acute kidney injury (AKI) in 

patients with cirrhosis and ascites. Recently, the International 

Club of Ascites (ICA) proposed new criteria for HRS, loosening 

some of the stricter criteria in order to enhance diagnostic 

sensibility. However, HRS diagnosis still requires the exclusion 

of certain triggers of AKI to ensure “hepatic” etiology of AKI. 

Aims: To assess survival of patients with HRS according to the 

new ICA-HRS guidelines as compared with patients with “specifically-triggered” 

AKI. Methods: A cohort of 497 consecutive 

patients with cirrhosis and ascites was longitudinally screened 

for renal dysfunction. AKI and HRS were diagnosed according 

to new ICA criteria. Specific triggers for severe (grade 2/3) AKI 

episodes that are considered exclusion criteria for HRS were 

recorded (sAKI). Transplant-free survival (TFS) of HRS and sAKI 

was compared using log-rank test. Results: Among all patients, 

64 cases of HRS and 124 cases of sAKI were recorded. No 

differences were found regarding patient characteristics or TFS: 

Median TFS was 24 days in HRS (IQR 4-405 days) vs. 16 days 

(3-217) in sAKI (p=0.364). sAKI with manifest hypovolemia as 

trigger was associated with the shortest TFS [n=70; 8 (3-40) 

days], followed by preceding surgery or intervention [n=6; 

TFS: 32 (3-109) days] and infections as triggers [n=30; TFS: 

44 (2-187) days]. Conversely, patients with acute-on-chronic 

renal failure [n=7; TFS: 250 (14-end of follow-up) days] and 

nephrotoxic trigger [n=11; TFS: 217 (7-387) days] showed a 

favorable TFS. Conclusion: Patients with HRS according to the 

new ICA guidelines and sAKI showed similar characteristics 

and outcomes. However, since the specific cause of sAKI significantly 

influences prognosis, identifying the trigger of sAKI 

represents an important clinical priority in cirrhotic patients 

with ascites. 

Disclosures: 



Simona Bota - Speaking and Teaching: Janssen Pharmaceutica, Bristol-Myers 

Squibb 

Wolfgang Sieghart - Grant/Research Support: Bayer Schering Pharma, Bayer 

Schering Pharma, Bayer Schering Pharma, Bayer Schering Pharma; Speaking 

and Teaching: Bayer Schering Pharma, Bayer Schering Pharma, Bayer Schering 

Pharma, Bayer Schering Pharma 











The following authors have nothing to disclose: Theresa Bucsics, Philipp Schwabl, 

Bernhard Scheiner, Arnulf Ferlitsch


271 

Chronic Rifaximin Therapy Decreases the Risk and 

Severity of Hepatorenal Syndrome in Cirrhosis 

Tien S. Dong, Andrew I. Aronsohn, Nancy Reau, K. Gautham 

Reddy, Helen S. Te; Medicine, University of Chicago, Chicago, IL 

BACKGROUND: The intestinal bacterial flora plays an important 

and complex role in patients with cirrhosis. Studies in alcohol-related 

cirrhotic patients showed that rifaximin reduced the 

frequency of acute kidney injury (AKI) and hepatorenal syndrome 

(HRS). AIM: To determine the effect of long-term rifaximin 

use on the renal function of cirrhotic patients. METHODS: 

A retrospective chart review was performed to identify cirrhotic 

patients who had at least two visits in the Liver Clinic from 

1/1/11 to 12/31/14. The study group consisted of patients 

who were on rifaximin for ≥90 days, who were then matched 

by age, gender, and baseline MELD score to a control group. 

Patients with a diagnosis of hepatocellular carcinoma and 

renal replacement therapy (RRT) at baseline were excluded. 

Demographic and laboratory data, concurrent midodrine use, 

incidence of AKI, HRS, need for RRT, infections, frequency of 

hospitalizations and length of stay (LOS) were collected. AKI 

was defined as an increase in serum Cr (SCr) by >0.3 mg/ 

dl within 48 hours or an increase in SCr >1.5 folds above 

baseline (International Society of Nephrology criteria). Data 

was censored at the last follow-up, initiation of RRT, death, or 

liver transplant, and analyzed using logistic regression, oneway 

ANOVA, and Pearson’s chi-squared test with the Stata 

software. RESULTS: 88 rifaximin cases were identified and 

matched to 88 control cases. Age, gender, race, duration of 

follow up, baseline MELD score, SCr, and GFR, and etiology 

of cirrhosis were not significantly different between the two 

groups. There were more patients on chronic midodrine (>90 

days use) in the rifaximin group as compared to control (17 % 

vs 4.5%, p= 0.008). There was no significant difference in the 

frequency of infections, deaths, liver transplants, hospitalizations 

or mean LOS between the two groups. Despite a similar 

frequency of AKI between the two groups, there was a trend 

towards less likelihood for HRS as a cause for AKI while controlling 

for midodrine use (OR 0.272 [0.067-1.086], p=0.06). 

In addition, the rifaximin group also had a lower likelihood of 

having at least one HRS episode (OR 0.257 [0.0696-0.950], 

p=0.04) and lower likelihood of requiring RRT (OR 0.257 

[0.067-0.950], p=0.04). The number needed to treat to prevent 

one episode of RRT is 10. CONCLUSIONS: Chronic use 

of rifaximin is associated with a decreased risk of HRS and a 

decrease likelihood of requiring RRT for AKI in a general population 

of cirrhotic patients. This finding should be validated in 

a prospective study in a larger patient population. 

Disclosures: 

Nancy Reau - Advisory Committees or Review Panels: Jannsen, Merck, AbbVie, 

Intercept, Salix, BMS, Jannsen; Grant/Research Support: Merck, Gilead, BMS, 

AbbVie, Jannsen, BI 

K. Gautham Reddy - Advisory Committees or Review Panels: AASLD Transplant 

Hepatology Pilot Steering Committee, ACG Training Committee, Program Director’s 

Caucus Steering Committee, Gilead, Inercept, Bristol-Myers Squibb; Grant/ 

Research Support: Intercept, Ocera, Merck, Lumena 

Helen S. Te - Advisory Committees or Review Panels: BMS; Grant/Research 

Support: Abbvie, Conatus, BMS 

The following authors have nothing to disclose: Tien S. Dong, Andrew I. Aronsohn 

272 

Ascites Neutrophil Gelatinase-Associated Lipocalin 

(NGAL) Identifies Spontaneous Bacterial Peritonitis and 

Predicts Mortality in Hospitalized Patients with Cirrhosis 

Elizabeth C. Verna 1 , Giuseppe Cullaro 1 , Grace Kim 1 , Mona Gossmann 

1 , Thresiamma Lukose 1 , Connie Kim 2 , Sofia Nigar 3 , Marcus 

Pereira 1 , Robert S. Brown 1 ; 1 Columbia University Medical Center, 

New York, NY; 2 Stony Brook University School of Medicine, Stony 

Brook, NY; 3 The Brooklyn Hospital Center, Brooklyn, NY 

Background: Neutrophil gelatinase-associated lipocalin 

(NGAL) levels in the urine strongly predict acute kidney injury 

(AKI) and mortality in patients with cirrhosis. Ascites NGAL 

(aNGAL) levels may identify peritonitis in patients on peritoneal 

dialysis but have not been tested in cirrhosis. We hypothesized 

that aNGAL level is a marker of spontaneous bacterial 

peritonitis (SBP) and mortality risk in hospitalized patients with 

cirrhosis. Methods: Hospitalized patients with cirrhosis and 

ascites undergoing diagnostic paracentesis were prospectively 

enrolled and followed until death or discharge. Exclusion criteria 

included secondary peritonitis, history of transplantation 

and active colitis. NGAL was measured in the ascites, serum 

(sNGAL) and urine (uNGAL) by ELISA (Bioporto, Denmark). 

aNGAL level was evaluated as a marker of SBP (defined as 

ascites absolute neutrophil count >250) and predictor of inpatient 

mortality. Results: 79 patients were enrolled in this ongoing 

study. The median age was 58.4, 59% male gender, 53% 

non-Hispanic white, 21% Hispanic, and 11% black. 44% had 

HCV as their primary liver disease, 35% alcohol and 10% 

NAFLD. The median MELD was 20, 26% had AKI, defined as 

an increase of 0.3 mg/dL from baseline, and median (range) 

follow up was 11 (3-374) days. 16 patients (20%) had SBP. 

Baseline characteristics were similar between subjects with and 

without SBP. Median (IQR) aNGAL was significantly higher 

in patients with SBP compared to those without SBP [297.0 

(365.0) v. 155.0 (162.5) ng/mL, p= 0.008]. In ROC analysis, 

aNGAL had an AUC of 0.72 (95% CI 0.59-0.86). Median 

sNGAL (454.5 v. 409.7 ng/mL, p=0.41) and uNGAL (133.6 

v. 99.3 ng/mL, p=0.07) were similar between groups. aNGAL 

correlated relatively well with sNGAL (r=0.74) and less so with 

uNGAL (r=0.40). 8 (10%) patients died in the hospital. aNGAL 

(OR 1.02 per 10 units, p=0.02), MELD (1.13, p=0.02), SBP 

(OR 4.91, p=0.04) and bacteremia (OR 6.2, p=0.02) were 

predictive of in hospital mortality. In the final multivariable 

model, aNGAL (OR 1.03 per 10 units, p=0.01) remained 

predictive of in hospital mortality, controlling for SBP (OR 9.05, 

p=0.03) and AKI (8.82, p=0.04). Conclusions: aNGAL level 

may be a biomarker of peritonitis in hospitalized patients with 

ascites, and importantly, a predictor of short term in hospital 

mortality even controlling for SBP and AKI. Larger studies are 

needed to validate these findings. 

Disclosures: 

Elizabeth C. Verna - Advisory Committees or Review Panels: Gilead; Grant/ 

Research Support: Salix, Merck 


The following authors have nothing to disclose: Giuseppe Cullaro, Grace Kim, 

Mona Gossmann, Thresiamma Lukose, Connie Kim, Sofia Nigar, Marcus Pereira


273 

Use of non-selective beta blockers in cirrhotic patients 

with ascites is associated with increased risk of acute 

kidney injury at hospital admission 

Juliana R. de Carvalho, Rodrigo Luz, Henrique Sergio M. Coelho, 

Cristiane Villela-Nogueira, Renata M. Perez; Hepatology Service, 

Federal University of Rio de Janeiro, Rio de Janeiro, Brazil 

Introduction Use of non-selective beta blockers (NSBB) in 

patients with advanced cirrhosis has recently become an issue 

of debate, with concerns about their safety and possible association 

with complications such as acute kidney injury (AKI). 

Objective The aim of this study was to evaluate the association 

between use of NSBB and the presence of AKI in the first 48 

hours of hospital admission in cirrhotic patients with ascites. 

Methods Hospitalizations of cirrhotic patients with ascites in 

an university hospital were retrospectively reviewed. AKI at 

hospital admission was defined as a difference of at least 0.3 

mg/dL in two serum creatinine values obtained in the first 48 

hours of admission. Association between use of NSBB and AKI 

was evaluated by logistic regression analysis. Results This study 

included 328 hospitalizations of cirrhotic patients with ascites, 

with a mean age of 59 (SD: ±12.7) years. Men represented 

59% of cases. At admission, 133 patients (41%) were classified 

as Child B and 195 (59%) as Child C. History of previous 

gastrointestinal bleeding episode was present in 144 patients 

(44%) and 200 (61%) were on NSBB. AKI at hospital admission 

occurred in 196 patients (60%), being present in 66% of 

patients on use of NSBB and 50% of those not on use of the 

drug (p=0.004). In Child B patients, AKI occurred in 61% of 

those on NSBB and 51% of those without the drug (p=0.24). 

In Child C patients, a significant association between NSBB 

use and AKI was observed (69% vs 49%; p = 0.006). The 

variables age, gender, previous episode of gastrointestinal 

bleeding, previous episode of spontaneous bacterial peritonitis, 

history of diabetes mellitus, history of arterial hypertension, 

use of diuretics, use of NSBB, Child-Pugh score at admission 

and serum sodium at admission were included in a logistic 

regression model. In the final model, variables independently 

associated to AKI in the first 48 hours of admission were: age 

(OR: 1.03; 95% CI: 1.00 – 1.05; p=0.004), serum sodium at 

admission (OR: 0.95; 95% CI: 0.92 – 0.99; p=0.014) and 

use of NSBB (OR: 1.91; 95% CI: 1.20 – 3.06; p=0.007). Conclusion 

In Chid C cirrhotic patients with ascites, use of NSBB 

is associated with increased risk of AKI at hospital admission. 

Use of these drugs in this population should be reassessed. 

Disclosures: 

The following authors have nothing to disclose: Juliana R. de Carvalho, Rodrigo 

Luz, Henrique Sergio M. Coelho, Cristiane Villela-Nogueira, Renata M. Perez 

274 

Slow, continuous low-dose albumin and furosemide 

infusion (SCLAFI) mobilizes large ascites safely in 

decompensated liver cirrhosis 

Gaurav Pande; gastroenterology, Sanjay gandhi post graduate 

institute of medical sciences, Lucknow, India 

Background: Graded increase of oral diuretics has been the 

standard therapy for mobilizing large ascites in decompensated 

liver cirrhosis. Large volume paracentesis (LVP) with or without 

albumin infusions has been the commonest rescue therapy. 

Both are sought with complications. Aim: To study the efficacy 

and safety of low-dose, continuous, infusionof furosemide with 

albumin, administered according to a response-guided protocol 

in patients with cirrhosis and large ascites. Methods: All cirrhotic 

patients with large ascites on maximally tolerated dirutics 

and creatinine


SC and UO criteria. We defined oliguria as transient if patient 

had UO stage 1 and persistent if patient had UO stage 2 or 

3. Of the 3458 patients with CLD included in the study, 2997 

(86.7%) had cirrhosis. AKI occurred in 2854 (82.5%) patients. 

Hospital mortality for patient with AKI vs no AKI was 21.2% vs 

5% (p


(


failure of baseline mean arterial pressure to increase by 10 

mm Hg. Central venous pressure was checked every four hours 

with a goal of 4-10 mm Hg. Additional intravenous albumin 

(25 g) was given if goals were not met and furosemide (120 

mg) was administered when goals were exceeded. Progressive 

improvement in serum Cr led to weaning of NE. Patients were 

considered treatment failures if serum Cr continued to increase 

on protocol or renal replacement therapy was initiated. Results: 

A total of 42 patients were included. Prior to NE infusion, the 

median creatinine was 3.1 mg/dL and the median MELD score 

was 33 points. The median albumin dose was 93.5 g prior to 

NE therapy and the median duration of therapy was 6.5 days. 

New cardiac ischemia or arrhythmia developed in 4/42 (9%) 

of treated patients resulting in cessation of NE therapy in 2 of 

those patients. Improvement in renal function without need for 

renal replacement therapy was seen in 20/42 (48%) patients. 

The median Cr in responders went from 3.45 to 2.4 mg/dL. Of 

the responders, 15/20 (75%) were discharged from the hospital 

(N=8) or successfully bridged to liver transplantation (N=7), 

while 5/20 (25%) died or were transitioned to hospice for 

reasons not related to worsening renal function. Conclusions: 

NE infusion plus intravenous albumin appears to be a relatively 

safe and effective therapy for type 1 HRS. In this analysis, we 

were able to improve or stabilize renal function to the point of 

liver transplantation or discharge from the hospital in 48% of 

patients meeting strict criteria for type 1 HRS, a disorder historically 

associated with a median survival of about 2 weeks. 

Disclosures: 

The following authors have nothing to disclose: Crit T. Richardson, Michael K. 

Porayko 

280 

Clinical Utilization of Serum Albumin to Ascitic Fluid 

Albumin Gradient and Ascitic Fluid Protein Concentration 

in Pediatric Ascites 

Saachi Nangia 1 , Thammasin Ingviya 3 , Pavis Laengvejkal 2 , Ann 

O. Scheimann 1 , Wikrom Karnsakul 1 , Hejab Imteyaz 4 , Alexandra 

Vasilescu 4 ; 1 Pediatrics, Johns Hopkins University School of Medicine, 

Baltimore, MD; 2 Neurology, Texas Tech University Health 

Sciences Center, Lubbock, TX; 3 Environmental Health Sciences, 

Bloomberg School of Public Health, Baltimore, MD; 4 Pediatric Gastroenterology, 

Ft Myers, FL 

Background: Abdominal paracentesis and ascitic fluid (AF) 

analysis is performed in children with ascites to rule out infection 

or cancer. Serum albumin to ascitic fluid albumin gradient 

(SAAG) is frequently used in adults to distinguish portal 

hypertension (PHT) when SAAG is ≥1.1g/dL. High AF protein 

(>2.5 g/dL) is reported in cases with congestive heart failure 

or congestive hepatopathy (CH), hepatic vein outflow obstruction 

(HVOO) while low AF protein ( 2.5 

g/dL; uniquely presenting with intrahepatic mass and reactive 

mesothelial cells on AF cytopathology. Of 7 HVOO&CH 

cases only a BCS case with hepatic GVHD and liver fibrosis 

had AF protein < 2.5 g/dL. An idiopathic case with SAAG of 

0.6 and AF protein 2.3 g/dL led to suspect cirrhosis (APRI > 

1). None of 21 cases had definite SBP. Conclusion: Our data 

showed consistency with existing literature that cases with PHT 

either from cirrhosis, HVOO, or CH had SAAG ≥1.1 g/dL 

except for one case with peritoneal dialysis. SAAG < 1.1 g/dL 

was observed in cases with NS, serositis, PLE and peritoneal 

metastasis. AF protein has its value to reconfirm PHT types and 

potential causes of ascites along with SAAG and APRI values. 

Disclosures: 

The following authors have nothing to disclose: Saachi Nangia, Thammasin 

Ingviya, Pavis Laengvejkal, Ann O. Scheimann, Wikrom Karnsakul, Hejab 

Imteyaz, Alexandra Vasilescu 

281 

Multicenter retrospective analysis of the Model for Endstage 

Liver Disease (MELD) and proton pump inhibitors 

to predict in the first episode of spontaneous bacterial 

peritonitis 

Sakkarin Chirapongsathorn 1,2 , Rashid Khan 3 , Ashwani K. Singal 4 , 

Patrick S. Kamath 1 ; 1 Gastroenterology and Hepatology, Mayo 

Clinic, Rochester, Rochester, MN; 2 Gastroenterology, Phramongkutklao 

Hospital and College of Medicine, Royal Thai Army, Bangkok, 

Thailand; 3 Gastroenterology and Hepatology, The University 

of Texas Medical Branch (UTMB) in Galveston, Galveston, TX; 

4 Gastroenterology and Hepatology, University of Alabama, Birmingham, 

AL 

Background: Spontaneous bacterial peritonitis (SBP) is a common 

infection in patients with cirrhosis. The Model for Endstage 

Liver Disease (MELD) is a prospectively developed and 

validated chronic liver disease severity scoring system to predict 

survival. At present, there are no data on its predictive ability 

on the first episode of SBP occurrence. Recent studies show 

controversial results regarding an increased risk of developing 

SBP in cirrhotic patients on proton pump inhibitors (PPI). The 

aim of the study was to investigate the relationship between 

MELD score and PPI therapy and the development of the first 

episode of SBP in cirrhotic patients in a multicenter study. Methods: 

A retrospective case-control study was performed. Seven 

hundred fifteen patients with a diagnosis of cirrhosis were identified 

from 3 tertiary referral centers. All medical records were 

manually reviewed by investigators to confirm the diagnosis 

of cirrhosis and SBP. Cirrhotic patients with a first episode 

of SBP (n=247 case) were matched to cirrhotics with ascites 

but without SBP (n=468 control). Every case was matched to 

2 controls for age, gender and race within each institution. 

Baseline MELD score was calculated within a month prior 

to SBP diagnosis for cases and within a month prior to any 

decompensation for controls. The time point for PPI usage was 

90 days before SBP diagnosis. Results: Proportion of patients 

with SBP increased with increasing MELD scores. For each unit 

increase in MELD, the odds ratio (OR; 95% CI) for development 

of SBP was 1.18 (1.14-1.21). Among patients with MELD score


greater than 10, 20 and 30, OR for development of SBP was 

4.62 (2.86-7.82), 7.9 (5.18-12.27) and 11.03 (4.92-28.1) 

respectively. MELD score cut-off level ≥ 17 had an area under 

the receiver operating characteristic curves (AUROC) 0.76 with 

sensitivity 63% and specificity 79%. PPI use was independently 

associated with development of SBP (OR 2.87, CI 1.78-4.69). 

There was an interaction between MELD score and PPI use, but 

MELD score was a stronger risk. Conclusion: MELD score and 

PPI therapy are associated with development of the first episode 

of SBP. Prospective studies are needed to validate these 

data and to determine the optimal MELD score above which 

patients would benefit from primary prophylaxis of SBP. 

18 and 24 months. Overall the included studies had a low risk 

of bias, except for two observational studies in which the risk 

was judged as moderate. Conclusions: Based on moderate 

quality evidence, the use of NSBBs was not associated with a 

significant increase in the risk of all-cause mortality in cirrhotic 

patients with ascites, including refractory ascites. 

Subgroup analyses of 12 studies included in meta-analysis 

Multivariate Logistic regression analyses of predictors the first 

episode of SBP 

Disclosures: 


The following authors have nothing to disclose: Sakkarin Chirapongsathorn, 

Rashid Khan, Ashwani K. Singal 

282 

Effect of Beta-Blockers on Survival in Patients with Cirrhosis 

and Ascites: A Systematic Review and Meta-analysis 

Sakkarin Chirapongsathorn 1,2 , Nelson Valentin 1 , Fares Alahdab 

3,4 , Chayakrit Krittanawong 5 , Patricia J. Erwin 6 , Mohammad 

H. Murad 3,7 , Patrick S. Kamath 1 ; 1 Gastroenterology and Hepatology, 

Mayo Clinic, Rochester, Rochester, MN; 2 Gastroenterology, 

Phramongkutklao Hospital and College of Medicine, Royal Thai 

Army, Bangkok, Thailand; 3 Robert D and Patricia E Kern Center 

for the Science of Health Care Delivery, Mayo Clinic, Rochester, 

Rochester, MN; 4 Knowledge and Evaluation Research Unit, Mayo 

Clinic, Rochester, Rochester, MN; 5 Cardiovascular Disease, Mayo 

Clinic, Rochester, Rochester, MN; 6 Mayo Clinic Libraries, Mayo 

Clinic, Rochester, Rochester, MN; 7 Preventive, Occupational and 

Aerospace Medicine, Mayo Clinic, Rochester, Rochester, MN 

Backgrounds: Recent studies have suggested that non-selective 

beta-blockers (NSBBs) are associated with negative impact 

on survival in patients with cirrhosis and refractory ascites. 

However, other studies showed contradictory findings and suggested 

that NSBBs are beneficial in these patients. We performed 

a systematic review and meta-analysis to evaluate the 

effect of NSBBs on all-cause mortality. Methods: We conducted 

a comprehensive search of MEDLINE, Embase, Web of Science 

and Scopus databases through January 2015 and manually 

reviewed the literature. Attempts were made to contact 

the corresponding authors of the relevant studies for additional 

information when needed. Trial-specific risk ratios (RRs) were 

calculated and pooled using random-effect model meta-analysis. 

Between-study heterogeneity was assessed using the I 2 

statistic. The quality assessment of included studies and publication 

bias were assessed. Result: We included 4 randomized 

control trials and 8 observational studies in which 1,206 deaths 

were reported in 2,486 patients with ascites. NSBBs users did 

not have an increase in all-cause mortality compared to NSBBs 

nonusers with overall ascites (RR, 0.94; 95% CI, 0.6-1.46; P 

= 0.77); non-refractory ascites (RR, 0.97; 95% CI, 0.45-2.09) 

or refractory ascites (RR, 0.86; 95% CI: 0.47-1.57; P = 0.63). 

Results were similar in RCTs and observational studies. Use of 

NSBBs was not associated with increased mortality at 6, 12, 

Disclosures: 



Nelson Valentin, Fares Alahdab, Chayakrit Krittanawong, Patricia J. Erwin, 

Mohammad H. Murad 

283 

Increasing incidence and cost, but decreasing mortality 

in patients with hepatorenal syndrome: a study of the 

National Inpatient Sample 2005-2011 

Albert Do 1 , Ghideon Ezaz 2 ; 1 Internal Medicine, Yale-New Haven 

Hospital, New Haven, CT; 2 Internal Medicine, Beth Israel Deaconess 


Background: Hepatorenal syndrome (HRS) is significant in 

patients with cirrhosis, carrying a high morbidity and mortality 

risk. However, its burden on inpatient hospital care presently 

is unclear. We report the incidence, costs, and mortality 

trends of HRS in the American medical inpatient setting. Materials 

and Methods: The Nationwide Inpatient Sample contains 

discharge-level inpatient hospitalizations comprising 20% of 

non-government hospitals in the United States. We determined 

incidence rates, mortality rates, hospital length of stay, and 

hospital costs (adjusted to 2011 dollar terms to adjust for inflation) 

from 2005 to 2011 for patients with HRS (ICD-9 code 

572.4). Results: 153,057 cases of HRS were identified during 

2005-2011, comprising a cohort with mean age 57.6 years, 

primarily males (64.1%) of white race (55.9%), with public 

insurance payers (58.1%), and hospitalized at a teaching hospital 

(54.9%). Table 1 summarizes HRS-associated trends in 

incidence, mortality and cost, showing increased disease incidence. 

Overall hospital mortality was 34.4%, with decreasing 

mortality (43.8% in 2005 to 31.2% in 2011). $16.3 billion 

was spent in this time, with an annual expenditure increase 

from $1.4 billion to $3.5 billion. However, hospital length 

of stay did not change during this time period (range 10.1 to 

11.9 days). 5,268 patients (3.4% of all patients) received a 

liver transplant during their hospital stay. Conclusions: From 

2005 to 2011, the incidence and costs of HRS have increased.


However, associated mortality has decreased and hospital 

length-of-stay has not changed. Further studies are needed to 

assess the reasons for decreased mortality, as well as measures 

to decrease costs associated with hospital stay. 

Table 1. Incidence, mortality, and costs of patients with hepatorenal 

syndrome, NIS sample 2005-2011 

had the highest bilirubin (27mg/dl) compared to ATN alone 

(18 mg/dl) p


is similar to previous reports. Guidelines recommend a paracentesis 

to evaluate for SBP in patients admitted with ascites, 

but do not address evaluation for SBP in outpatients. In outpatients, 

the need for routine ascitic fluid analysis in asymptomatic 

patients is an area of discussion based on a reported low 

prevalence of SBP. This study suggests that similar to inpatients, 

all asymptomatic outpatients undergoing paracentesis should 

routinely have ascitic fluid analyzed to assess for SBP. 

Disclosures: 

Maria Sjogren - Speaking and Teaching: Gilead, Bristol Myers 

The following authors have nothing to disclose: Dustin Albert, Manish B. Singla, 

Frank Cheng, Dawn Torres 

286 

Is Midodrine, Octreotide, and Albumin still worthwhile 

therapy for Hepatorenal Syndrome Type 1 (HRS1)? 

Katherine Hahn 1 , Amen Javaid 1 , Pooja Singhal 2 , Franca B. Barton 

3 , James H. Lewis 2 ; 1 Internal Medicine, Georgetown University 

Hospital, Washington, DC; 2 Georgetown University, Washington, 

DC; 3 The EMMES Corporation, Rockville, MD 

Introduction: HRS1 is a fatal complication of advanced cirrhosis, 

with mortality rates as high as 80% within 2 weeks 

of diagnosis. Current treatment options in the United States 

include vasoconstrictors (e.g.: midodrine [M], octreotide [O], 

and vasopressin), often combined with albumin [A], transjugular 

intrahepatic portosystemic shunt (TIPS), renal replacement 

therapy (RRT), and liver transplantation (LT). This retrospective 

study analyzes the outcomes of 137 HRS1 patients treated 

with MOA as primary therapy or as a bridge to LT. Methods: A 

retrospective cohort study evaluated HRS1 patients treated with 

MOA at Georgetown University Hospital over a 6 year period 

(Jan 2009 – Dec 2014). The demographics, underlying liver 

disease, creatinine levels, MELD scores, RRT requirements, and 

LT rates of these MOA patients were collected. The primary end 

point was survival to hospital discharge. Secondary outcomes 

included survival after RRT and survival with/without LT. The 

effects of patient characteristics on survival were tested with a 

Cox proportional hazards model. The relationship between LT 

and survival was analyzed using logistic regression. Results: 

From 2009-2014, a total of 238 patients were treated with 

MOA. Of these, 101 patients did not meet the diagnostic 

criteria for HRS1 and were excluded. Of the remaining 

137 patients, 70 (51%) had SBP, 73 (52.9%) received RRT, 

but only 34 (24.8%) met criteria to undergo LT. The overall 

survival to discharge was 33%. Among those who survived, 

57.1% had received LT and 42.8% had not. Factors showing 

a significant effect on survival were LT (p=0.015), MELD score 


288 

Acute kidney injury in cirrhotic patients undergoing contrast-enhanced 

computed tomography 

Roberto Filomia 1 , Sergio Maimone 1 , Carlo Saitta 1 , Luca Visconti 5 , 

Simona Caloggero 4 , Antonio Bottari 4 , Alessia Pitrone 4 , Angela 

Alibrandi 3 , Irene Cacciola 1 , Gaia Caccamo 1 , Domenica Vadalà 1 , 

Carmine G. Gambino 1 , Maria Stella Franzè 1 , Giovanni Raimondo 

1 , Giovanni Squadrito 1,2 ; 1 Clinical and Molecular Hepatology, 

University Messina, Messina, Italy; 2 Human Pathology, 

University of Messina, Messina, Italy; 3 Department of Economical, 

Business and Environmental Sciences and Quantitative Methods, 

Univeristy of Messina, Messina, Italy; 4 Department of Biomedical 

Sciences and Morphological and Functional Imaging, University of 

Messina, Messina, Italy; 5 Internal Medicine, Unit Of Nephrology, 

Messina, Italy 

Introduction: Contrast-induced acute kidney injury (CI-AKI) 

following intravenous contrast medium administration is one 

of the most common causes of hospital-acquired acute kidney 

damage. Aim: To investigate the incidence and possible 

predisposing factors of CI-AKI in cirrhotic patients undergoing 

contrast-enhanced computed tomography (CECT). Patients 

and Methods: We analysed 393 consecutively hospitalized 

cirrhotic patients none of whom had active bacterial infection, 

glomerular filtration rate (GFR)


pressure (mPAP)


was observed between the two groups of patients as far as 

demographic, clinical, hemodynamic, and laboratory characteristics. 

No treatment was provided for Grade 1 ascites in 

the study. During the follow up twenty-two patients developed 

Grade 2 or 3 ascites, 14 in Group A (14,7 %) and 8 in Group 

B (25.0%, p = N.S.). In only 4 out of 14 patients of Group A, 

Grade 1 ascites was detected before they developed Grade 2 

or 3 ascites. The mean time for the development of Grade 2 

or 3 ascites was shorter in Group B than in Group A, but the 

difference was not statistically significant (17,00 ± 11,06 versus 

32,29 ± 27,59 months, p = N.S.). In nineteen patients in 

Group B (59.3%), Grade 1 ascites disappeared during the follow 

up. At univariate analysis, the finding of Grade 1 ascites, 

MELD, spleen diameter and the ratio platelets/spleen diameter 

were found as potential predictors of Grade 2 or 3 (Table). At 

multivariate analysis only the ratio platelets/spleen diameter 

was found to be the only independent predictor of Grade 2 or 

3 ascites (HR = 0,997, 95%, CI : 0,994; 0,999, p = 0,0018 

per el/mm 3 /mm of increase). In conclusion, the presence of 

Grade 1 ascites is neither a precursor nor an independent 

predictor of Grade 2 or 3 ascites in patients with cirrhosis. 

Thus, Grade 1 ascites does not require any treatment, including 

sodium dietary restriction. 

Disclosures: 

Paolo Angeli - Advisory Committees or Review Panels: Sequana Medical 

The following authors have nothing to disclose: Marta Tonon, Salvatore Piano, 

Matjaz Grbec, Chiara Pilutti, Silvano Fasolato, Antonietta Romano, Sara Montagnese, 

Filippo Morando, Massimo Bolognesi, Marialuisa Stanco, Giancarlo 

Bombonato, Silvia Rosi, Elisabetta Gola, Antonietta Sticca 

293 

Validation of Ascites-specific Patient Reported Outcome 

Questionnaires for Patients with Cirrhosis 

Patrick S. Kamath 3 , Myrte Neijenhuis 3 , Tom J. Gevers 2 , Thomas 

Atwell 1 , Joost Drenth 2 , Tim Gunneson 3 ; 1 Division of Gastroenterology, 

Hepatology & Internal Medicine, Mayo Clinic College of 

Medicine, Rochester, MN; 2 Gastroenterlogy and Hepatology, 

Radboud University Nijmegen Medical Centre, Nijmeten, Netherlands; 

3 Division of Gastroenterology and Hepatology, Mayo 

Clinic, Rochester, MN 

Background and aim: Since no treatment improves long-term 

survival, treatment of ascites should focus on symptom relief 

and prevention of complications. No patient reported outcomes 

have been used as endpoints in clinical trials in cirrhotic ascites. 

We aimed to identify a sensitive tool to measure symptoms 

in cirrhotic ascites. Methods: We identified 12 ascites-specific 

symptoms (fullness, lack of appetite, early satiety, nausea, 

abdominal pain, back pain, dyspnea, limited mobility, fatigue, 

insomnia, dissatisfaction with size of abdomen and sexual intimacy 

problems) based on literature, patient interviews (n=8) 

and clinician survey (n=8) to develop an ascites-specific questionnaire 

(Ascites-Q). We administered Ascites-Q; Japanese 

Ascites Symptom Inventory-7 (ASI-7, developed for cirrhotic 

ascites); FACIT-ascites index (developed for malignant ascites); 

and quality of life (QoL) VAS-scale in cirrhotic patients undergoing 

large volume paracentesis (target sample size n=90). 

Scores were transformed to a 0-100 scale with higher scores 

indicating more symptoms. To identify ascites-specific symptoms, 

we compared using the Mann Whitney U test scores 

of patients with refractory ascites prior to paracentesis with 

scores in cirrhotic patients with no ascites recruited at the Mayo 

Clinic between January - May 2015. Correlations of QoL VASscale 

and ascites questionnaires were calculated (Spearman). 

Responsiveness of questionnaires to detect changes in symptoms 

was assessed by comparing scores at baseline and 7 

days post paracentesis (paired T-test). Results: We included 32 

patients with refractory ascites requiring large-volume paracentesis 

(59% male, mean age 61 yr, MELD-score 15 and median 

paracentesis volume 3600 mL; and 61 patients without refractory 

ascites (controls) (76% male, mean age 57 yr, MELD-score 

11). Patients with refractory ascites scored higher than controls 

on all symptoms of Ascites-Q and ASI-7, except for Ascites-Q 

symptoms back pain (p=0.096) and fatigue (p=0.228). There 

was no difference between patients and controls in 6/13 FAC- 

IT-ascites index symptoms (insomnia, limited mobility, nausea, 

vomiting, polyuria and emotional distress). QoL correlated with 

Ascites-Q and ASI-7 (r=.497, p=0.008 and 0.471, p=0.013), 

but not with FACIT-ascites index (r=0.332, p=0.090). Scores 

of Ascites-Q, (-11.53 ± 18.874, p=0.013), ASI-7 (-35.46, ± 

29.22, p


2 BA, 2 HVOO and 1 CH and HE diagnosed in 10 BA and 

7 HVOO. HRS and HE therefore were not included in logistic 

regression analysis as a result of samll sample size. Conclusion: 

BA group was younger and had higher propability of developing 

significant ascites and splenomegaly. Variceal bleeding 

was a complication in BA group presenting in a younger age 

range. Longitudinal study is needed for more detailed analysis 

to understand the natural history of children with PHT from 

diffrernet groups. - 

Disclosures: 

The following authors have nothing to disclose: Thammasin Ingviya, Ann O. 

Scheimann, Wikrom Karnsakul 

295 

CA-125 significance in cirrhosis and its correlation with 

disease severity 

Raja G. Reddy Edula, Yucai Wang, Serban A. Moroianu, Vivek 

A. Lingiah, Phoenix Fung, Maliha Ahmad, Lizza Bojito-Marrero, 

Nneoma O. Okoronkwo, Nikolaos Pyrsopoulos; Medicine, Rutgers 

New Jersey Medical School, Newark, NJ 

Background: Serum cancer antigen 125(CA125) is a tumor 

marker traditionally used in the diagnosis and monitoring of 

ovarian cancer. However it is reported to be elevated in a 

variety of non-neoplastic conditions like cirrhosis and congestive 

heart failure. Its prevalence, degree of elevation, correlation 

with severity of disease based on objective markers like 

MELD score, Child’s-Pugh classification and degree of ascites 

in cirrhosis is unknown. Aim: To evaluate the prevalence 

and significance of elevated CA125 in patients with cirrhosis 

and its correlation with objective markers of disease severity. 

Methods: This is a prospective study of 172 adult patients with 

cirrhosis due to any etiology after obtaining CA125 serum 

analysis at the time of enrollment. Demographics, etiology of 

cirrhosis, MELD score, Child’s-Pugh classification, degree of 

ascites, serum CA125 and other variables were collected. Statistical 

analysis including descriptive statistics, linear regression, 

one-way ANOVA and Student’s t-test was performed. 

Results: Etiology of cirrhosis is outlined in Table-1. Elevated 

CA125 levels were noted in 147(85%) of the study population 

with a mean value of 379. No difference was observed in 

elevated mean CA125 between male(411) and female(324) 

patients(P=0.207). Patients with higher MELD had higher 

CA125 level (P= 0.001). Subjects without ascites (N=35) had 

a mean CA125 level of 36, mild ascites(N=29): 218 and moderate 

ascites(N=108): 534(P


297 

Impact of SNP rs2187668 in HLA-DQA1 on clinical 

presentation and treatment response in patients with 

type-1 autoimmune hepatitis 

Albert Stättermayer 1 , Michael Eder 1 , Sandra Beinhardt 1 , Karin 

Kozbial 1 , Clarissa Freissmuth 1 , Friedrich Wrba 2 , Michael Trauner 1 , 

Peter Ferenci 1 , Harald Hofer 1 ; 1 Gastroenterology & Hepatology, 

Medical University of Vienna, Vienna, Austria; 2 Clinical Pathology, 

Medical University of Vienna, Vienna, Austria 

Background/aim: Autoimmune hepatitis (AIH) is a chronic 

inflammatory liver disease with limited understanding of etiopathogenesis. 

Recently, a genome-wide association study 

(GWAS) identified a single nucleotide polymorphism (SNP, 

rs2187668) in the HLA-DQA1 gene, which is strongly associated 

with AIH type-1. We evaluated the impact of this SNP on 

clinical presentation and treatment response in patients with 

type-1 AIH. Methods: One-hundred and seven patients with 

type-1 AIH (female: 83 [77.6%], mean age: 43.2 [CI95%: 

39.9-46.6] years), who reached a probable or definite score 

according to the simplified or revised scoring system were evaluated. 

SNP rs2187668 was investigated by real-time-PCR in 

all patients with AIH and in a control group of 97 healthy 

subjects. Results: Sixty-one (57.0%) patients with AIH had 

rs2187668 genotype GG, 39 (36.4%) were heterozygous 

and 7 (6.5%) had genotype AA. Minor allele frequency (MAF) 

of the risk allele (A) was 24.8% and was significantly higher 

than in the control group (MAF: 7.2%; p150 vs.


minimal and discordant, the aims of our study were to: 1) 

investigate HPSE expression in HSCs during their activation; 

2) evaluate HPSE activity in the plasma of patients with liver 

fibrosis. Methods. HSCs were isolated from rat liver and cultured 

on plastic to induce activation. HPSE expression, together 

with markers of HSCs transdifferentiation, were evaluated by 

real-time PCR at different days after isolation. HPSE protein 

amount was evaluated by immunofluorescence (IF). Utilizing 

an ELISA method, HPSE activity was measured in the plasma 

from 15 healthy subjects and 45 patients diagnosed with autoimmune 

chronic liver diseases (15 AIH, 17 PBC, 13 PSC); 

HPSE activity was correlated with fibrosis staging assessed by 

elastography. Results. After 15 days on plastic, HSCs resulted 

activated with the increase of alpha-SMA, fibronectin and TGFbeta 

expression with respect to 7 days cultured cells. Upon activation, 

HPSE mRNA was significantly 1.94-fold overexpressed 

in 15 days vs 7 days cultured cells. HPSE increase was confirmed 

by IF. HPSE plasma activity was significantly higher in 

patients with mild, significant and severe fibrosis compared 

to the control group (0.32±0.04, 0.25±0.03, 0.21±0.03 

respectively vs 0.06±0.01, p


(0.05 ng/mL -0.16 ng/mL), (p=0.003 and p=0.02, respectively)). 

Following in vitro activation, PD-1 was up-regulated 

3.3 fold (p=0.0019) on T cells from AIH patients compared 

with HC (1.5 fold). Conclusion: Present study supports an 

increased PD-1 mediated immune-regulation in AIH patients. 

Furthermore our results support the hypothesis about the presence 

of exhausted T cells in autoimmune diseases. 

Disclosures: 

Henning Gronbaek - Grant/Research Support: Novartis, Ipsen 

The following authors have nothing to disclose: Kristian Aarslev, Anders Dige, 

Stinne Greisen, Martin Kreutzfeltd, Niels Jessen, Hendrik V. Vilstrup, Bent Deleuran 

302 

Autoimmune hepatitis type 1 in Dutch elderly versus 

younger patients 

Martine A. Baven-Pronk 1 , Joanne J. van Silfhout 1 , Aad P. van 

den Berg 2 , Henk R. van Buuren 3 , Carin M. Van Nieuwkerk 4 , Bart 

van Hoek 1 ; 1 Gastroenterology and Hepatology, Leiden University 

Medical Center, Leiden, Netherlands; 2 Gastroenterology and 

Hepatology, University Medical Center Groningen, Groningen, 

Netherlands; 3 Gastroenterology and Hepatology, Erasmus Medical 

Center, Rotterdam, Netherlands; 4 Gastroenterology and Hepatology, 

Free University Medical Center, Amsterdam, Netherlands 

Background and Aims: Studies on elderly patients with autoimmune 

hepatitis (AIH) are limited, of small sample size and 

conflicting when it comes to mode of presentation, treatment 

and treatment outcome. The aim of this study was to investigate 

differences in baseline characteristics, laboratory features, 

histology, concomitant autoimmune disease, treatment, 

treatment adverse effects and outcome in elderly patients with 

AIH type one (age > 60 years at presentation) compared to 

younger patients with AIH type one (age < 60 years at presentation) 

and compare them to existing data to provide more 

insight into AIH in the elderly patient. Patients and Methods: 

All patients from 4 academic centres with probable or definite 

AIH type I according to the International AIH Group criteria 

were included. As much data as possible was retrospectively 

retrieved by chart review. Primary endpoints were defined as 

remission, liver transplantation or liver related death. Secondary 

endpoints were defined as differences in biochemistry and 

serology, symptoms, mode of presentation, concurrent autoimmune 

diseases, initial and maintenance treatment regimens, 

number of switches of therapy, adverse effects of treatment, 

achievement of remission, episodes of loss of remission, number 

of relapses, cirrhosis at presentation and progression of cirrhosis. 

Results: A total of 359 patients were included, 286 (80%) 

younger patients, 73 (20%) elderly patients. The young group 

presented with significantly higher serum alanine aminotransaminase 

(p


304 

Carcinogenesis in autoimmune hepatitis type 1 patients 

with a long-term follow-up in Japan 

Teruko Arinaga-Hino 1 , Tatsuya Ide 1 , Ichiro Miyajima 1 , Kei Ogata 1 , 

Reiichiro Kuwahara 1 , Keisuke Amano 1 , Kazunori Noguchi 2 , Kunihide 

Ishii 3 , Kunitaka Fukuizumi 4 , Koji Yonemoto 5 , Takuji Torimura 

1 ; 1 Division of Gastroenterology, Department of Medicine, 

Kurume Univ.School of Medicine, Kurume, Japan; 2 Omuta City 

Hospital, Omuta, Japan; 3 Asakura Medical Association Hospital, 

Asakura, Japan; 4 National Kyushu Medical Center, Fukuoka, 

Japan; 5 The Biostatistics Center, Kurume University School of Medicine, 

Kurume, Japan 

Backgrounds/Aims: The risk of carcinogenesis in autoimmune 

diseases is high and has been attributed to immunological 

abnormalities, the use of immunosuppressive agents, and/or 

chronic inflammation. The aim of the present study was to determine 

the incidence and risk of carcinogenesis in patients with 

autoimmune hepatitis (AIH) type 1 in a large-scale population 

with a long-term follow-up in Japan. Methods: Two hundred 

and fifty-six patients diagnosed with AIH between Apr. 1978 

and Mar. 2014 were enrolled (F/M = 226:30; mean age 

at diagnosis AIH, 57.7 yrs). They had no malignancy before 

or within 1 yr of being diagnosed with AIH. A person-year 

calculation was performed for AIH patients, and the numbers 

of expected events were clarified using data from “A Study 

of 28 Population-based Cancer Registries for the Monitoring 

of Cancer Incidence in Japan (MCIJ) Project (2010)” in order 

to determine the standard incident rate (SIR) of each type of 

malignancy. Biochemical data regarding carcinogenesis and 

its background factors were also examined using a proportional 

hazards model, adopting the Kaplan-Meier method. The 

present study was approved by the proper Ethics Committee. 

Results: The mean observation period was 8.2 yrs (The person 

years at risk were 2,107; Females; 1,878, Males; 229). 

Twenty-seven patients developed malignancy (F:M = 24:3). 

The mean observation period of carcinogenesis from AIH diagnosis 

was 10.3 yrs. The number of patients with hepatobiliary 

cancer (Ca) was 11 (hepatocellular carcinoma; 10, cholangiocarcinoma; 

1), gastric Ca; 3, pancreatic Ca; 2, oral/pharyngeal 

Ca; 2, colorectal Ca; 2, breast Ca; 1, lung Ca; 1, acute 

myeloid leukemia; 1, malignant lymphoma 1, and malignant 

melanoma; 1. The overall SIR for malignancies in AIH was 

significantly high at 2.04 (95% CI 1.34-2.96), and was particularly 

high among females at 2.49 (95% CI 1.60-3.71). SIR for 

hepatobiliary Ca was 14.14 (95% CI 7.05-25.30), and was 

markedly high for females at 21.83 (95% CI 10.45-40.16). 

SIR for oral/pharyngeal Ca was significantly high for females 

at 14.61 (95% CI 1.64-52.77). No significant differences 

were observed in SIR among the other types of malignancies 

examined. The risk factors for overall carcinogenesis at the 

diagnosis of AIH were identified as low levels of alanine aminotransferase 

(p=0.0053), a low platelet count (p=0.0087), 

and liver cirrhosis (p=0.0067). No significant differences were 

observed among the treatment regimens, such as prednisolone, 

azathioprine, and ursodeoxycholic acid. Conclusion: The 

risk of malignancies was generally high among AIH patients. 

The risk of carcinogenesis needs to be carefully considered in 

patients with AIH. 

Disclosures: 

The following authors have nothing to disclose: Teruko Arinaga-Hino, Tatsuya 

Ide, Ichiro Miyajima, Kei Ogata, Reiichiro Kuwahara, Keisuke Amano, Kazunori 

Noguchi, Kunihide Ishii, Kunitaka Fukuizumi, Koji Yonemoto, Takuji Torimura 

305 

Immunity in HLA-DR4 expressing healthy and autoimmune 

hepatitis induced NOD mice 

Muhammed Yuksel 1,2 , Xiaoyan Xiao 1,3 , Kathie Béland 4 , Pascal 

Lapierre 4 , Fernando Alvarez 4 , Isabelle Colle 2 , Yun Ma 5 , Li Wen 1 ; 

1 Endocrinology, Yale university, New haven, CT; 2 Internal medicine, 

Ghent university hospital, Ghent, Belgium; 3 Department of 

Nephrology, Shangdong University, Shangdong,., China; 4 Division 

of Gastroenterology, Hepatology and Nutrition, Sainte-Justine 

University Hospital, Montreal, QC, Canada; 5 Institute of Liver studies, 

Kings College London, London, United Kingdom 

Autoimmune hepatitis (AIH) is a chronic progressive inflammatory 

liver disease, characterized by interface hepatitis, positivity 

for autoantibodies and hyper-gammaglobulinemia. There are 

two types of AIH, type-1 (AIH-1) being characterized by seropositivity 

for anti-smooth muscle and/or anti-nuclear (SMA/ 

ANA), and type-2 (AIH-2) by positivity for anti-liver kidney microsomal 

type 1 (anti-LKM1) and/or anti-liver cytosol type 1 

(anti-LC1) autoantibodies. Cytochrome P4502D6 (CYP2D6) 

has been defined as the antigenic target of anti-LKM1 and 

formiminotransferase cyclodeaminase (FTCD) as the target 

for anti-LC1. The development of AIH is linked to the Human 

Leukocyte Antigen (HLA) alleles, -DR3 for AIH-1 and -DR7 for 

AIH-2. However, the role of HLA-DR4 has been controversial, 

being the second disease predisposition gene in adults with 

AIH-1 and protective gene in pediatric patients. In this study, 

we investigated the role of HLA-DR4 transgene, expressed on 

non-obese diabetic (NOD) mice (DR4), healthy (baseline) mice 

and also after the AIH induction by immunization with the DNA 

coding for human CYP2D6/FTCD fusion protein. Immunization 

of wild type (WT) NOD and DR4 mice leads to a sustained 

mild elevation of alanine aminotransferase (ALT), development 

of anti-LKM1/anti-LC1, and chronic immune cell infiltration, 

mild parenchymal fibrosis and some plasma cell infiltration. 

Although these parameters were more slightly pronounced in 

DR4 mice than in WT control mice, the pathogenic role of 

HLA-DR4 remains uncertain. However, livers from DR4 mice 

contained less regulatory T cell (Tregs) which had decreased 

PD-1 expression and a dysfunction to suppress alloreaction. 

We also showed that, T cells and antigen presenting cells, 

such as macrophages and dendritic cells, were already activated 

in non-immunized naïve (healthy) DR4 mice compared to 

naïve WT NOD. These results show that HLA-DR4 molecule is 

involved in the spontaneous activation of the immune system, 

facilitating the induction of AIH. 

Disclosures: 

Isabelle Colle - Advisory Committees or Review Panels: Janssen; Grant/Research 

Support: Bayer; Patent Held/Filed: Trombogenics; Speaking and Teaching: BMS 

The following authors have nothing to disclose: Muhammed Yuksel, Xiaoyan 

Xiao, Kathie Béland, Pascal Lapierre, Fernando Alvarez, Yun Ma, Li Wen 

306 

Autoimmune Hepatitis Disproportionately Affects People 

of Color 

Michele M. Tana 1,2 , Edward W. Holt 3 , Robert J. Wong 4 , Justin L. 

Sewell 1 , Mandana Khalili 1 , Jacquelyn J. Maher 1,2 ; 1 Department of 

Medicine, Division of Gastroenterology, University of California, 

San Francisco, San Francisco, CA; 2 UCSF Liver Center, University 

of California, San Francisco, San Francisco, CA; 3 Department of 

Transplantation, Division of Hepatology, California Pacific Medical 

Center, San Francisco, CA; 4 Department of Gastroenterology 

and Hepatology, Alameda Health System, Oakland, CA 

Autoimmune hepatitis (AIH) is an uncommon disease that 

affects patients of all ethnicities, and there have been reports 

that its natural history varies in patients of different ethnic back-


grounds. There is insufficient evidence about the prevalence 

of AIH among people of color.We sought to determine the 

racial/ethnic distribution of AIH cases in an ethnically diverse, 

urban, county hospital system. We performed an observational 

cohort study. A consecutive sample of patients with a diagnosis 

of AIH or AIH-PBC overlap syndrome per AASLD guidelines 

from September 2013 to April 2015 was included in 

the study. Demographic, clinical, laboratory, radiographic, 

and histologic data were collected. The characteristics of the 

cohort were compared with the published data on the patient 

population served by the integrated hospital system using the 

chi-squared test. The characteristics of different ethnic groups 

were compared using univariate and multivariate analysis. 23 

patients with AIH (6 had overlap syndrome) were seen in the 

hepatology clinic over the 20-month period. AIH occurred in 

significantly fewer white patients than would be expected if the 

prevalence were equal among ethnic groups, given the demographics 

of this integrated hospital system (Table, p=0.03). 

The median age at diagnosis was 49 years (range 18-91) and 

18 (78%) were female. At diagnosis, the median total bilirubin 

was 3.3 mg/dL, ALT was 483 U/L, IgG was 1995 mg/dL, INR 

was 1.2, albumin was 3.8 g/dL, and platelet count was 177 

k/uL. 19 patients had complete liver biopsy results, and ten 

(53%) had fibrosis stage ≥2, with four (21%) having advanced 

(stage 3-4) fibrosis. When stratified by Latino ethnicity, Latinos 

did not differ significantly from non-Latinos in terms of age at 

diagnosis, baseline labs, or severity of liver fibrosis. A two-predictor 

model that adjusted for age found no significant association 

between Latino ethnicity and fibrosis stage. AIH was 

significantly more prevalent among people of color (especially 

Latinos) suggesting that AIH may not affect all ethnic groups 

equally. Although the disease characteristics did not appear 

to differ by ethnicity, this disparity may be related to genetic, 

socioeconomic, environmental, or comorbid medical factors. 

Disclosures: 

Mandana Khalili - Consulting: Gilead Sciences Inc; Grant/Research Support: 

Gilead Sciences INc, Bristal Myer Squibb 

Jacquelyn J. Maher - Consulting: Durect 

The following authors have nothing to disclose: Michele M. Tana, Edward W. 

Holt, Robert J. Wong, Justin L. Sewell 

307 

Outcomes and mortality of de novo autoimmune hepatitis 

after liver transplantation 

Aishwarya Kuchipudi 1 , Ryan Morton 3 , Sarah Y. Chan 3 , Dilip 

Moonka 2 , Kimberly Ann Brown 2 , Syed M. Jafri 2 ; 1 Internal Medicine, 

Henry Ford Hospital, Detroit, MI; 2 Gastroenterology, Henry 

Ford Hospital, Detroit, MI; 3 Wayne State University School of Medicine, 

Detroit, MI 

Aim: To evaluate the incidence and outcomes of de novo Autoimmune 

Hepatitis (AIH) in patients with history of Orthotopic 

Liver Transplantation (OLT). Methods: We performed a retrospective 

chart review of 1343 OLT patients from 2000-2015. 

We identified patients with histopathological evidence of de 

novo AIH. Patients with biopsy features consistent with AIH 

pre-OLT and those with Hepatitis C were excluded from analysis. 

For each patient with de novo AIH, we identified 2 controls 

wherever possible, matched for etiology of liver disease 

and date of transplant (+/- 18 months). We then compared 

the two groups on factors including intra-operative or post-operative 

complications, immunosuppression levels at specific 

time points, development of fibrosis/cirrhosis, progression 

to re-transplant and mortality at 1, 3 & 5 years respectively. 

Results: Of 1343 patients who received an OLT, 6 developed 

de novo AIH. The etiology of initial liver failure was alcohol in 

3 of these patients; and steatohepatitis, cryptogenic hepatocellular 

carcinoma, primary sclerosing cholangitis in one patient 

each. We identified a total of 11 matched controls for these 

patients. Mean donor age was 45 years in the de novo group 

and 40 years in the control group (p=0.62). 2 patients in the 

de novo group and 4 in the control group developed biliary 

stricture post-operatively. One of the control patients developed 

post-op cholestasis. Mean Tacrolimus levels at week 1; week 

12, 6 months and 1 year were not statistically different. Mean 

Tacrolimus levels at week 4 was 10 in the de novo AIH group 

and 5.41 in the control group (p= 0.02). At 1 year, levels 

were 8.53 in the de novo group and 7.0 in the control group 

(p=0.61). The average time to development of de novo AIH 

was 1096 days (range 31-2822 days). Mean total prednisone 

used was 2697 mg in the de novo group and 1711 mg in the 

control group (p=0.60). Three de novo patients were noted 

to have fibrosis at the time of diagnosis (2 with severe fibrosis 

and 1 with mild fibrosis) and 1 control patient developed 

mild fibrosis (p=0.09). 1 de novo patient developed cirrhosis 

(788 days), none of the controls did. None of the patients 

received a re-transplantation. Survival at 1 year was 100% in 

the de novo group and 82% in the control group. Survival at 3 

years was 80% and 82% respectively. Survival at 5 years was 

40% and 82% respectively (p=0.34). Two deaths in the control 

group and one death in the de novo group were attributed to 

infection. Conclusion. De novo AIH is relatively rare post-OLT. 

It might be associated with a higher potential for significant 

fibrosis and increased mortality. 

Disclosures: 

Dilip Moonka - Advisory Committees or Review Panels: Gilead; Grant/Research 

Support: Bristol-Myers Squibb; Speaking and Teaching: AbbVie, Gilead 

Kimberly Ann Brown - Advisory Committees or Review Panels: CLDF, Merck, 

BMS, ABBVIE, Gilead, Gilead, Janssen, Salix; Consulting: Blue Cross Transplant 

Centers; Grant/Research Support: CLDF, Gilead, Exalenz, CDC, BMS, Hyperion, 

Merck; Speaking and Teaching: ABBVIE, Gilead, CLDF 

The following authors have nothing to disclose: Aishwarya Kuchipudi, Ryan 

Morton, Sarah Y. Chan, Syed M. Jafri


308 

Tacrolimus or Mycophenylate mofetil as a second-line 

therapeutic options in patients with Autoimmune Hepatitis: 

An international multicentre observational study. 

Cumali Efe 5 , Hannes Hagström 3 , Rahima A. Bhanji 2 , Niklas F. 

Müller 1 , Qixia Wang 4 , Tugrul Purnak 5 , Luigi Muratori 6 , Mårten 

Werner 7 , Hanns-Ulrich Marschall 8 , Paolo Muratori 6 , Fulya Gunsar 

9 , Daniel Klintman 10 , Albert Parés 11 , Alexandra Heurgué-Berlot 

12 , Thomas D. Schiano 13 , Xiong Ma 4 , Aldo J. Montano-Loza 2 , 

Thomas Berg 1 , Ersan Ozaslan 14 , Eric M. Yoshida 15 , Staffan Wahlin 

3 ; 1 Universitätsklinikum Sektion Hepatologie, Klinik für Gastroenterologie 

und Rheumatologie., Leipzig,, Germany; 2 University of 

Alberta Division of Gastroenterology and Liver Unit., Alberta, AB, 

Canada; 3 Gastroenterology and Hepatology, Karolinska Institutet, 

Karolinska University Hospital., Stockholm, Sweden; 4 Department 

of Hepatology, Shanghai University of Traditional Chinese Medicine, 

Longhua Hospital, Shanghai,, China; 5 Department of Gastroenterology, 

Hacettepe University,, Ankara, Turkey; 6 Department of 

Clinical Medicine, Alma Mater Studiorum, University of Bologna,, 

Bologna, Italy; 7 Departments of Medicine, Sections for Hepatology 

and Gastroenterology, University Hospital of Skåne,, Malmö, 

Sweden; 8 Department of Molecular and Clinical Medicine, Sahlgrenska 

Academy, University of Gothenburg,, Gothenburg, Sweden; 

9 Department of Gastroenterology, Ege University, Bornova,, 

Izmir, Turkey; 10 Department of Molecular and Clinical Medicine, 

Lund University of Hospital,, Lund, Sweden; 11 Liver Unit, Hospital 

Clínic University of Barcelona C/ Villarroel 170 08036, Barcelona, 

Spain; 12 Department of Hepato-Gastroenterology, CHU 

Reims,, Reims, France; 13 Division of Liver Diseases, The Mount 

Sinai Medical Center, New York,, NY; 14 Department of Gastroenterology, 

Numune Research and Education Hospital,, Ankara, Turkey; 

15 Division of Gastroenterology University of British Columbia 

and Vancouver General Hospital., Vancouver, BC, Canada 

Background: Autoimmune hepatitis (AIH) is a well-recognized 

immune mediated liver disease. Eighty percent (80 %) of 

patients respond to standard therapy, including corticosteroids 

alone or combination with azathioprine (AZA). However, this 

therapy is not tolerated or insufficient to control disease activity 

in 20 % of patients. Second-line treatment with Tacrolimus (Tac) 

or Mycophenylate mofetil (MMF) has been described in only 

small case series. The aim of this study was to evaluate the outcome 

of Tac and/or MMF therapy in patients with AIH who are 

resistant or intolerant to standard immunosuppression. Patients 

and methods: This multi-centre retrospective chart review study 

evaluated data of AIH, including overlap with PBC or PSC 

patients, treated with Tac and/or MMF. Initial treatment was 

corticosteroids ±AZA. The patients were categorized as complete 

responders to initial therapy (Group-I), partial responders 

(Group-II) and lack of responders (Group-III). The IAIHG criteria 

were used for the diagnosis of AIH. Results: 163 patients (121 

female), mean age of 37 (9-76) years were included. Patients 

were treated with Tac (n=71) and MMF (n=92) a median of 

16 months (1-252 months) after standard immunosuppression. 

MMF and Tac resulted in complete or partial response in all 

of group-I patients. In group-II and group-III, Tac induced complete 

or partial response in 75% (36/45) vs 50 % (23/46) of 

MMF patients (p=0.003). Median follow up duration was 42.5 

(1-2430) months after MMF and/or Tac therapy. Five and 10 

years follow up rates were 37 % (34/92) and 19.6 % (18/92) 

for MMF patients and 51% (36/71) and 14% (10/71) for Tac 

patients. Combination of Tac and MMF was given to only 6 

patients, with 4 responding to therapy. Eleven patients were 

eventually transplanted. MMF and/or Tac were discontinued 

due to side effects in 7 patients including, 4 Tac and 3 MMF. 

Conclusions: MMF and Tac are generally effective and well tolerated 

in standard treatment resistant or intolerant AIH patients. 

There is no significant differences in outcomes between MMF 

and Tac treatments in AIH patients who responded to conventional 

therapy. However, Tac may be superior in AIH patients 

who showed incomplete or non-response to standard immunosupression. 

Disclosures: 









The following authors have nothing to disclose: Cumali Efe, Hannes Hagström, 

Rahima A. Bhanji, Niklas F. Müller, Qixia Wang, Tugrul Purnak, Luigi Muratori, 

Mårten Werner, Paolo Muratori, Fulya Gunsar, Daniel Klintman, Albert Parés, 

Thomas D. Schiano, Xiong Ma, Aldo J. Montano-Loza, Ersan Ozaslan, Eric M. 

Yoshida, Staffan Wahlin 

309 

Killer cell immunoglobulin-like receptors and their HLA 

class I ligands in autoimmune hepatitis: a possible 

pathogenetic role ? 

Simona Onali 2 , Roberto Littera 1 , Carlo Carcassi 1 , Luca Secci 2 , 

Sara Lai 1 , Rita Porcella 1 , Sara Cappellini 2 , Claudia Salustro 2 , 

Cinzia Balestrieri 3 , Giancarlo Serra 3 , Maria Conti 3 , Teresa 

Zolfino 4 , Francesco Figorilli 2 , Michele Casale 2 , Stefania Casu 2 , 

Maria Cristina Pasetto 2 , Laura Matta 2 , Rosetta Scioscia 2 , Lucia 

Barca 2 , Luchino Chessa 2,3 ; 1 Medical Genetics, Department of 

Medical Sciences “M.Aresu”, University of Cagliari, Cagliari, 

Italy; 2 Center for the Study of Liver Diseases, Department of Medical 

Sciences “M.Aresu”, University of Cagliari, Cagliari, Italy; 

3 Department of Internal Medicine, Azienda Ospedaliero-Universitaria, 

Cagliari, Cagliari, Italy; 4 S.S.D. Coordinamento Trapianti di 

fegato, Azienda Ospedaliera Brotzu, Cagliari, Italy 

Background: Natural killer (NK) cells are widely distributed in 

hepatic tissue, but their role in the pathogenesis of autoimmune 

hepatitis (AIH) has not been fully estabilished yet. Killer cell 

immunoglobulin receptors (KIRs) represent the major family of 

cell surface receptors that inhibit and activate NK cells. Aim: 

To assess whether different type of KIRs could be involved in 

pathogenesis of AIH. Methods: We analyzed all consecutive 

Sardinian outpatients with type I AIH, referred to the Liver Unit 

of University of Cagliari (Italy) between January and October 

2014. AIH was diagnosed according to the International Autoimmune 

Hepatitis Group Revised Original Scoring System and 

AASLD guidelines. Patients with type II AIH, overlap syndrome 

and other chronic liver diseases were excluded. The immunogenetic 

characteristics of AIH patients were compared to those 

of 221 healthy individuals extracted from the Sardinian bone 

marrow donor registry. High resolution (4 digits) typing of HLA- 

A, B, C and DR loci and 15 KIRs gene loci was performed in 

both patients and controls. Subjects were divided into 2 groups 

according to homozygosity for KIR haplotype A (KIR genotype 

AA), heterozygosity or homozygosity for KIR haplotype B (KIR 

genotype Bx). The study was approved by local ethics committee 

and informed consent was obtained from all participants. 

Results. 114 subjects were included: median age 55 years 

(range 17-80), 87.7% females, median duration of disease 6 

years (range 0.6-23). The frequency of activating KIR gene, 

KIR2DS1, was significantly higher in AIH patients compared 

to controls (57% vs 43%, p=0.03, HR=1.7, 95% CI=1.0-2.7), 

as well as KIR2DL1+/KIR2DS1+/HLA-C2+ (47.3% vs 35.7%, 

p=0.04, HR=1.6, 95% CI=1.0-2.6). When we considered the 

age of onset of the autoimmune hepatitis, we found that activating 

KIR gene KIR2DS1 was more frequent in patients diag-


nosed at younger age. Conclusions: Based on our findings, the 

activating KIR gene KIR2DS1 is highly expressed in patients 

with autoimmune hepatitis, suggesting its potential involvement 

in pathogenesis of type I AIH. However further studies are 

needed to fully understand the role of KIR2DS1 as marker for 

AIH. 

Disclosures: 

Teresa Zolfino - Board Membership: Abvie; Grant/Research Support: Bayer; 

Speaking and Teaching: Bristol 

Luchino Chessa - Board Membership: Abbvie; Speaking and Teaching: BMS, 

Jannsen 

The following authors have nothing to disclose: Simona Onali, Roberto Littera, 

Carlo Carcassi, Luca Secci, Sara Lai, Rita Porcella, Sara Cappellini, Claudia 

Salustro, Cinzia Balestrieri, Giancarlo Serra, Maria Conti, Francesco Figorilli, 

Michele Casale, Stefania Casu, Maria Cristina Pasetto, Laura Matta, Rosetta 

Scioscia, Lucia Barca 

310 

Possible involvement of activating follicular helper T 

cells in autoimmune hepatitis 

Naruhiro Kimura, Satoshi Yamagiwa, Hiroki Honda, Toru Setsu, 

Kentaro Tominaga, Hiroteru Kamimura, Masaaki Takamura, 

Shuji Terai; gastroenterology and hepatology, Niigata university, 

Niigata, Japan 

BACKGROUND: There is an increasing interest in the role of 

follicular helper T (Tfh) cells in autoimmunity from the perspective 

of both their role in breaking tolerance and their effects on 

disease progression. Dysregulated Tfh cells have been shown 

to be responsible for the induction of fatal autoimmune hepatitis 

(AIH) in mice that are unable to produce natural regulatory T 

cells after neonatal thymectomy and that are genetically devoid 

of the programmed cell death 1 (PD-1)-mediated signaling. 

However, the involvement of Tfh cells in the immune pathogenesis 

of AIH has not been fully characterized. AIM: To evaluate 

the numbers of different subsets of circulating Tfh and B cells 

and their potential role in the pathogenesis of AIH. METHODS: 

Heparinized peripheral blood was collected from 18 patients 

with AIH, 22 patients with chronic hepatitis B (CHB) and 21 

healthy volunteers (HC). Mononuclear cells were separated 

using the Ficoll gradient, and various surface markers were 

then investigated using flow cytometry. Cytokine production 

was investigated using peripheral blood Tfh cells after stimulation 

with phorbol myristate acetate (PMA) and ionomycin. We 

also investigated the distribution of CXCR5 + CD4 + cells in the 

liver using immunohistochemical staining. RESULTS: CXCR5 + 

CD4 + Tfh cells comprised 8.3% (median) (range 2.7-18.3), 

7.8% (4.1-13.5), and 8.1% (3.1-13.5) of the total T cells in the 

blood of patients with AIH, CHB, and HC, respectively. The Tfh 

cells were then classified into several subsets according to their 

expression of PD-1, inducible co-stimulator (ICOS) and CXCR3. 

We found a significant increase in the proportion of activated 

ICOS + Tfh cells in the blood (19.4%) and livers (24.3%) of 

patients with AIH compared with the proportion in the blood 

of the same patients after prednisolone (PSL) treatment (1.6%). 

The frequency of ICOS + Tfh cells was significantly decreased 

in the PSL-treated patients (median 22.6%, range 15.2-34.5) 

compared with the patients who were not treated with PSL 

(median 35.7%, range 26.0-56.2), although the levels of alanine 

aminotransferase (ALT) were decreased after treatments 

in both groups (17 ± 30.2 IU/l in PSL-treated and 15 ± 14.7 

IU/l in non-PSL-treated patients). CXCR5 + cells were detected 

among infiltrated lymphocytes in the portal area of the liver 

of patients with AIH. Furthermore, we confirmed that Tfh cells 

secreted IL-21 and IL-17 and expressed PD-1 after stimulation 

with PMA and ionomycin. CONCLUSIONS: Previous studies 

have reported that an increase in Tfh cells is associated with 

the activity of autoimmune diseases. Our results suggest that 

ICOS + Tfh cells may be associated with the disease activity of 

AIH. 

Disclosures: 

Shuji Terai - Speaking and Teaching: Otsuka Pharma. 

The following authors have nothing to disclose: Naruhiro Kimura, Satoshi 

Yamagiwa, Hiroki Honda, Toru Setsu, Kentaro Tominaga, Hiroteru Kamimura, 

Masaaki Takamura 

311 

Methotrexate therapy for Autoimmune Hepatitis 

James Haridy 1 , Amanda J. Nicoll 1,2 , Siddharth Sood 1 ; 1 Department 

of Gastroenterology and Hepatology, Royal Melbourne 

Hospital, Melbourne, VIC, Australia; 2 Department of Gastroenterology, 

Eastern Health, Melbourne, VIC, Australia 

Background: Autoimmune hepatitis (AIH) is characterised by 

typically diverse phenotypical manifestation and response to 

treatment. Corticosteroids and azathioprine are standard firstline 

therapies. There are limited options in patients who display 

poor response or intolerance to these medications. Methotrexate 

(MTX) is readily available, often employed in other autoimmune 

conditions, and has therefore been postulated as a 

possible second-line therapy. Evidence of efficacy has been 

limited to a small number of case-reports, despite more widespread 

anecdotal use. Aim: To assess the response to MTX in 

AIH. Method: A retrospective case-series was conducted using 

subjects located through the gastroenterology liver database of 

a tertiary referral centre. Subjects with a diagnosis of AIH were 

identified and diagnosis confirmed using the International Autoimmune 

Hepatitis Group criteria. Medical records were examined 

to determine medication history. Patients were included 

if they had prior treatment exposure to methotrexate (MTX) for 

their AIH. Baseline and follow-up data was collected to determine 

response to MTX over the initial 12-months of therapy. Follow-up 

was collected up to 3-years following initiation of MTX 

if available. Results: 11 patients (age 24 – 72 years; mean, 

56 years) were identified with confirmed AIH and MTX use. 

10/11 (90.9%) were female and 4/11 (36.4%) had biopsy 

or imaging evidence of cirrhosis prior to initiation of MTX. The 

most common indication for MTX was incomplete response 

to thiopurines (5/11, 45.5%). The median dose of prednisolone 

prior to initiating MTX was 8.0mg (range 0-15mg). 5/11 

(45.5%) of subjects ceased MTX within twelve months due to 

an adverse event, most commonly deterioration in liver function 

(range 1-12 months). 5/11 (45.5%) achieved or maintained 

complete remission at 12-months on MTX. These patients experienced 

a median decrease in prednisolone dosage of 5.3 mg, 

with a median dose of 3 mg (range 0 – 5mg) at 12-months. 

1/11 (9%) achieved a partial response at 12-months but 

later achieved complete remission on MTX monotherapy by 

36-months. 1/6 (16.7%) patients continuing MTX at 12-months 

were steroid free. Conclusion: Prior use of MTX in AIH has been 

limited to three individual case reports, however it is mentioned 

in guidelines as a possible alternative second-line therapy. In 

our case series, we show that MTX is safe, and that over 50% 

of patients are able to be maintained on it with good effect on 

liver function and minimal steroid use. MTX may have a role in 

treatment of AIH in a subset of patients who are refractory or 

intolerant of first-line therapies. 

Disclosures: 

Amanda J. Nicoll - Grant/Research Support: MSD; Speaking and Teaching: 

Bayer 

Siddharth Sood - Grant/Research Support: Qiagen USA 

The following authors have nothing to disclose: James Haridy


312 

Features and patterns of concurrent extrahepatic autoimmune 

diseases in patients with Autoimmune Hepatitis 

and Overlap/Variant Syndromes: Implications for diagnosis 

and management 

Guan Wee Wong, Andrew D. Yeoman, Michael A. Heneghan; 

Institute of Liver Studies, King’s College Hospital, London, London, 


Aim: The presence of concurrent extrahepatic autoimmune disease 

(CEAID) represents an important diagnostic criterion in 

the International Autoimmune Hepatitis Group (IAIHG) scoring 

system. We aim to describe the clinical patterns of CEAID in 

patients with AIH and Overlap Syndromes (OS). Methods: The 

medical records and prospectively accrued electronic database 

of 300 patients with definite AIH as defined by the revised 

IAIHG criteria were reviewed. We also interrogated the medical 

records of 28 patients defined as OS, 10 AIH/PBC and 

18 AIH/PSC. Patients with Autoimmune Sclerosing Cholangitis 

(AISC) were excluded. Results: A total of 34 CEAID diagnosis 

were identified in our 328 patients. Among them, 81 patients 

(24.7%) had one, 19 (5.8%) had two, 2 (0.6%) had three 

diagnosed CEAIDs. In total, 88/300 (29%) of AIH patients 

had CEAID. Autoimmune thyroid disease (AITD) was identified 

in 36 patients (12%) with hypothyroidism being more common 

(21/300, 7%) followed by symptomatic goitre (8/300, 2.7%) 

and hyperthyroidism (5/300, 1.6%). Prevalence of a-priori diabetes 

(non-steroid induced) was found in 13 patients (4.3%), 

with 12 patients (92%) being Type 1. Inflammatory bowel disease 

(IBD) was present in 8 patients (2.7%) with ulcerative colitis 

(UC) representing the underlying IBD diagnosis. Connective 

tissue disease (CTD) was identified in 28 patients (9%), with 

rheumatoid arthritis reported in 15 patients (5%) and other 

rarer CTDs including non-specific arthritis (5 patients, 1.6%), 

SLE/ Lupus and Sjogren’s syndrome (3 patients each, 1%), Raynaud’s 

and antiphospholipid syndrome (1 patient each, 0.3%). 

Autoimmune skin lesions were identified in 13 patients (4.3%) 

ranging from vitiligo (4 patients, 1.3%), alopecia (3 patients, 

1%), leucocytoclastic vasculitis (2 patients, 0.6%), to erythema 

nodosum, lichen planus and erythema multiforme (1 patient 

each, 0.3%). Miscellaneous conditions (n=13) were reported 

including idiopathic thrombocytopenia purpura, multiple sclerosis 

and myasthenia gravis. For OS, the pattern of AIH/PBC 

associated CEAID differed from pure AIH, with Sjogren’s syndrome 

being more representative (30%). Similarly, UC was 

more commonly found in patients with AIH/PSC overlap (33%). 

Moreover, positive family history of CEAID was identified in 

52/300 (17.3%) patients with AIH. Of these, 23/52 (44 %) 

had CEAID in their personal history. Conclusions: CEAIDs are 

common in AIH and OS. ATID, Sjogren’s syndrome and UC 

are the most common CEAIDs identified in our patients with 

AIH, AIH/PBC and AIH/PSC respectively. Early recognition of 

these features may be the first clue leading to subsequent early 

diagnosis and effective management of AIH and OS. 

Disclosures: 

The following authors have nothing to disclose: Guan Wee Wong, Andrew D. 

Yeoman, Michael A. Heneghan 

313 

The Need for Improved Liver Literacy in the US Population 

Tracy J. Mayne, Herbert Swanson; Intercept Pharmaceuticals, San 

Diego, CA 

Objective: To assess the level of awareness, knowledge, attitudes 

and behaviors surrounding liver health and liver disease 

in the general US population. Methods: We conducted online 

surveys from 1/6/15 – 1/12/15 in the GfK Knowledge Panel, 

a stratified probability sample representative of the population 

of US households. To correct for technology & income bias, 

households without a computer/internet were provided hardware/internet 

access to participate. We selected 511 respondents 

using random probability address-based sampling. Final 

data were weighted by age, region, race/ethnicity, education, 

and income according to the 3/14 US Census Current 

Population Survey. Margin of error was ±3.4%. Results: Most 

respondents do not perceive themselves to be at risk for liver 

disease. They do not think about or discuss it with friends, family 

or their physician. Almost half had some belief that a person 

can live without a liver. Most participants were unaware that 

liver function testing is part of routine bloodwork, and few 

discussed liver test results with their physician. These findings 

do not reflect general lack of health awareness or knowledge: 

91% of participants were aware of tests and values for blood 

pressure (91%), blood sugar (81%), cholesterol (79%), and 

BMI (69%), and respondents were much more likely to discuss 

these with their physician. Respondents reported greater likelihood 

of thinking and worrying about other diseases (weight, 

heart, breast, mental, prostate, colon and kidney) than liver. 

Patients reported more stigma associated with liver cirrhosis 

than with kidney disease, heart disease, cancer (colon, breast, 

prostate, or lung), diabetes, or reproductive health problems. 

Conclusions: In a representative US sample, awareness and 

concern about liver disease ranked low compared to other 

diseases. Participants report low levels of interaction with physicians 

about liver disease compared to other diseases, and 

were much less familiar with measures of liver health than (for 

example) cardiovascular. Massive public health campaigns in 

cardiovascular health have raised awareness and increased 

monitoring, screening & treatment. With increasing non-viral 

liver diseases, such as non-alcoholic fatty liver and steatohepatitis, 

these data point to a need for broad public health educational 

campaigns in liver disease. 

Disclosures: 


Herbert Swanson - Management Position: Intercept 

314 

Longitudinal alteration in health-related quality of 

life and its impact on clinical course in patients with 

advanced hepatocellular carcinoma who received 

sorafenib treatment 

Masako Shomura 1 , Tatehiro Kagawa 2 , Koichi Shiraishi 2 , Shunji 

Hirose 2 , Yoshitaka Arase 2 , Kota Tsuruya 2 , Sachiko Takahira 3 , 

Haruka Okabe 1 , Tetsuya Mine 2 ; 1 Department of Nursing, Tokai 

University School of Health Sciences, Isehara city, Japan; 2 Department 

of Gastoroenterology, Tokai University School of Medicine, 

Isehara, Japan; 3 Department of Nursing, University of Nagasaki, 

Nagasaki, Japan 

Purpose: This study aims to clarify longitudinal alteration in 

health related quality of life (HRQOL) in patients with advanced 

hepatocellular carcinoma (HCC) receiving sorafenib until 

death or treatment discontinuation, and to seek the HRQOL


domains associated with prognosis. Methods: We assessed 

HRQOL using SF-36 v2 TM every three months and calculated 

eight norm-based domain scores defined 50 points as national 

standards in consecutive patients with advanced HCC who 

received sorafenib. We prospectively analyzed the relationship 

among HRQOL, baseline characteristics, overall survival 

(OS) and treatment duration by Cox’s proportional hazards 

model. All subjects were given educational instructions and 

telephone follow-up. Results: Fifty-four patients participated in 

the study, comprising 42 males (78%) with median age of 71 

years, 23 (43%) with hepatitis C virus infection, 33 (61%) with 

Child-Pugh score 5, and 30 (56%) with TNM stage IV. Median 

OS and treatment duration were 9 and 5 months, respectively. 

Forty patients (74%) died. Thirteen patients receiving sorafenib 

over one year maintained all domain scores above 40 without 

significant decline throughout the treatment period. In contrast, 

physical functioning (PF), role physical, and vitality scores 

declined continuously and significantly, one year before death 

in 40 deceased patients. Child-Pugh score 5 and PF score 

^40 at baseline were significantly associated with longer OS 

by multivariate analysis. Social functioning (SF) score ^40 

was the only significant predictor for longer treatment duration 

(Figure). Conclusions: HRQOL was not impaired significantly 

in patients who received sorafenib treatment over one year. 

PF score ^40 and SF score ^40 at baseline was significantly 

associated with longer OS and longer treatment duration, 

respectively. Thus, HRQOL could be a valuable marker to predict 

the clinical course of HCC patients receiving sorafenib. 

Disclosures: 

The following authors have nothing to disclose: Masako Shomura, Tatehiro 

Kagawa, Koichi Shiraishi, Shunji Hirose, Yoshitaka Arase, Kota Tsuruya, Sachiko 

Takahira, Haruka Okabe, Tetsuya Mine 

315 

Interferon (IFN), Ribavirin (RBV), Duration of Treatment 

and Baseline Patient-Reported Outcomes (PROs) Predict 

Adherence to the New Regimens for Treatment of 

Chronic Hepatitis C (CH-C) 

Zobair M. Younossi 1,2 , Maria Stepanova 3 , Linda Henry 3 , Fatema 

Nader 1 , Sharon L. Hunt 3 ; 1 Center For Liver Disease, Department 

of Medicine, Inova Fairfax Hospital, Falls Church, VA; 2 Betty and 

Guy Beatty Center for Integrated Research, Inova Health System, 

Falls Church, VA; 3 Center for Outcomes Research in Liver Disease, 

Washington, DC 

BACKGROUND AND AIM: Patients’ experience (PROs) and 

treatment regimens may affect treatment adherence. Our aim 

was to assess the impact of clinical and PRO factors on adherence 

to different anti-HCV regimens. METHODS: PRO data (SF- 

36, CLDQ-HCV, FACIT-F, WPAI:SHP) from 13 multinational 

clinical trials of anti-HCV treatment were available. Treatment 

non-adherence was defined as


316 

TNF-alpha is the Most Consistent Predictor of Impairment 

of Mental Health-Related Patient-Reported Outcomes 

(PROs) in Patients with Chronic Hepatitis C (CH-C) 

Treated with Ledipasvir (LDV) and Sofosbuvir (SOF) with 

or without Ribavirin (RBV) 

Zobair M. Younossi 1,2 , Maria Stepanova 2 , James M. Estep 2 , Azza 

Karrar 2 , Bibiana Oe 2 , Elzafir Elsheikh 2 , Siddharth Hariharan 2 , 

Kazi Ahmed 2 , Patricia Tran 2 , Fatema Nader 2 , Linda Henry 2 , Ali 

A. Weinstein 2 , Naomi Lynn Gerber 1,2 ; 1 Center For Liver Disease, 

Department of Medicine, Inova Fairfax Hospital, Falls Church, 



Background and aim: HCV infection has been associated with 

changes in brain metabolites that may result directly from the 

virus or mediated by exposure to proinflammatory cytokines 

in the central nervous system. Our aim was to evaluate the 

association of cytokines and mental health PROs in HCV+ 

patients treated with LDV/SOF±RBV. Methods: We included 

100 CH-C GT1 patients (age 53.1±9.6, 57% male, 86% 

white, BMI of 28.5±5.6) treated with LDV/SOF (N=50) or 

LDV/SOF+RBV (N=50). All patients completed several validated 

PRO instruments. Mental health PROs were measured 

by the Role Emotional (RE), Mental Health (MH) and Mental 

Component Summary (MCS) of SF-36, Emotional Well-being 

(EWB) of FACIT-F, and Emotional domain (EMM) of CLDQ- 

HCV. Frozen serum was analyzed for interleukins (IL) 1, 1ra, 

8, 10, tumor necrosis factor-α (TNF-α), interferon-γ, monocyte 

chemotactic protein 1/chemokine ligand 2, granulocyte colony 

stimulating factor, vaso-endothelial and platelet-derived 

growth factors using Bio-Plex, ELISA, and enzymatic assays 

baseline, treatment week 12 and 4 weeks post-treatment. Relationships 

between fatigue and clinical and laboratory data 

were assessed by multiple regression analyses. Results: Patients 

treated with LDV/SOF+RBV had more significant anemia (10% 

vs 0%; p=0.02), more psychiatric (26% vs 10%; p=0.04) and 

skin (22% vs 6%, p=0.02) adverse events during treatment. 

After adjustment for pre-treatment history of anxiety, the most 

consistent independent biomarker predictor of lower mental 

health PROs during treatment was the level of serum TNF-α 

(p


318 

Uncertainty and Hepatitis C: A Correlational Study 

Examining Mishel’s Uncertainty in Illness Theory 

Humberto Reinoso; Nursing, Mercer University, Atlanta, GA 

Background: Hepatitis C is the most common blood-borne infection 

worldwide. It is estimated that some 5.3 million Americans 

are living with hepatitis C. Some studies predict up to 

7.1 million individuals affected including high-risk populations 

(Chak, Talal, Sherman, Schiff, & Saab, 2011). Individuals born 

between 1945 and 1965, those from the baby boomers generation, 

account for approximately three-fourths of all hepatitis C 

cases in the U.S. (Centers for Disease Control and Prevention 

[CDC], 2012). Purpose: The purpose of this study was to examine 

Mishel’s Uncertainty in Illness Theory among individuals 

with hepatitis C. The study focused on the relationship a set 

of variables prescribed by the model had on individual’s perception 

of uncertainty. Methods: A cross-sectional design was 

used to study the relationship between uncertainty and a set 

of variables prescribed by Michel’s Uncertainty in Illness Theory 

(i.e. educational attainment, healthcare authority figures, 

social network, years since diagnosis, treatment attempts, and 

treatment experience). Results: Data were collected over an 

8-week period from a convenience sample of participants born 

between 1945 and 1965, who self-identified as having hepatitis 

C (n = 146). The relationship between perceived uncertainty 

and a set of variables prescribed by the theory was investigated 

using Pearson product-moment correlation coefficients. 

Preliminary analyses were preformed to ensure no violation of 

the assumptions of normality, linearity, and homoscedasticity. 

Educational attainment (r = .2, p = < .05) as well as treatment 

experience (r = .27, p = < .05) shared a positive relationship 

with the individual’s perception of uncertainty. Social network 

(r = -.51, p = < .01) shared a strong negative or inverse relationship 

with participant’s perception of uncertainty. Furthermore, 

there was a strong positive relationship between the 

perception of uncertainty experienced by those individuals with 

hepatitis C and information provided by healthcare authority 

figures (r = .73, p = < .01). Conclusion: The results of this study 

suggest that the perception of uncertainty experienced by those 

individuals with hepatitis C may be influenced positively or 

negatively by information provided by their healthcare authority 

figures as well as their social network. Further research is 

recommended, in the form of a multiple regression analysis, of 

those variables identified of having strong relationships with an 

individual’s perception of uncertainty in order to identify their 

predictive ability. 

Disclosures: 

The following authors have nothing to disclose: Humberto Reinoso 

319 

Small Anti-Warburg Molecules Kill Hepatocarcinoma 

Cells by Opening Voltage Dependent Anion Channels 

and Promoting Mitochondrial Oxidative Stress 

David N. DeHart 1 , Monika Gooz 1 , John J. Lemasters 1,2 , Eduardo 

N. Maldonado 1 ; 1 Drug Discovery & Biomedical Sciences, Medical 

University of South Carolina, Charleston, SC; 2 Biochemistry & 

Molecular Biology, Medical University of South Carolina, Charleston, 

SC 

BACKGROUND: Enhanced aerobic glycolysis and suppressed 

mitochondrial metabolism characterize the Warburg metabolism 

of tumors. Flux of metabolites across mitochondrial outer 

membranes occurs through voltage dependent anion channels 

(VDAC). Dimeric α,β-tubulin closes VDAC in planar lipid 

bilayers (PNAS 105:18746) and limits ingress of respiratory 

substrates and ATP through VDAC that support mitochondrial 

membrane potential (ΔΨ) formation in cancer cells (Cancer 

Res. 70:10192). The small molecule erastin opens VDAC by 

antagonizing the inhibitory effect of tubulin on VDAC (JBC 

288:11920). Here, we hypothesized that small molecules that 

antagonize VDAC-tubulin interaction increase mitochondrial 

formation of reactive oxygen species (ROS), decrease glycolysis, 

and activate c-jun N-terminal kinase (JNK), leading to 

mitochondrial dysfunction, cell death and tumor growth inhibition 

in hepatocarcinoma (HCC). Our AIM was to evaluate 

the effects of VDAC-tubulin antagonists on mitochondrial ΔΨ, 

ROS and lactate generation, JNK activation and cell killing 

in HepG2 and Huh7 HCC cells and in a xenograft model 

of Huh7 cells. METHODS: Confocal/multiphoton fluorescence 

microscopy assessed ΔΨ (tetramethylrhodamine methylester) 

and ROS (chloromethyldichlorofluorescein [cmDCF], MitoSOX 

Red). JNK was assessed by Western blotting and cell killing 

by propidium iodide fluorometry. Subcutaneous xenografts of 

Huh7 cells were developed in nude mice. RESULTS: Erastin 

and small molecules X1 and X2 identified in a high-throughput 

screen increased ΔΨ and prevented mitochondrial depolarization 

caused by cytosolic high free tubulin. Increased ΔΨ 

was followed by mitochondrial depolarization occurring 1-2 

h after X1 and X2 and 3-4 h after erastin. Lactate generation 

after X1 decreased by 60%. ROS increased 30 min after X1 

and 60 min after X2 and erastin, as assessed with cmDCF 

and MitoSOX. The mitochondrially targeted antioxidant MitoQ 

blocked this ROS increase. Additionally, erastin caused JNK 

phosphorylation within 1 h. X1 and X2-dependent cell killing 

of HCC cells was blocked by the antioxidant N-acetylcysteine. 

By contrast, X1 and X2 caused


of CHOP and downstream target ATG5, an E3-like enzyme 

required for lipidation of LC3b, through short hairpin RNA 

repressed induction of autophagy and enhanced cell viability 

following saturated FFA exposure. Conversely, addition of 

chloroquine or bafilomycin A1, inhibitors of lysosome activity, 

further enhanced LC3b and P62 accumulation in combination 

with saturated FFA-induced cell death. Analysis of 

ultrastructural features and immunocytochemistry revealed that 

saturated FFA exposure perturbed proper fusion of autophagosomes 

with lysosomes. Together, these results suggest that 

exposure of hepatocytes to saturated FFA induce the UPR, and 

this stress response pathway enhances initiation of autophagy. 

Furthermore, saturated FFA block the flux of autophagy, which 

contributes to hepatoxicity. Co-treatment of saturated FFA with 

caspase inhibitor ZVAD-FMK, along with imaged localization 

of cleaved caspase 3, indicated that apoptosis was not playing 

a major role in hepatocyte cell death in this model system. 

However, treatment with saturated FFA or other inhibitors of 

autophagic flux led to an increase in proteasome activity in a 

CHOP dependent manner. It has been suggested that regulation 

of autophagy and the ubiquitin-proteasome pathway are 

closely linked, and often aberrant in diseases. This supports 

the idea that saturated FFA induces proteasome activity in a 

CHOP-dependent manner following inhibition of autophagic 

flux. To understand the relationship between the UPR and inhibition 

of autophagic flux in NASH patients, we assayed liver 

biopsy samples and found up-regulation of CHOP coincident 

with accumulation of LC3b and P62 in NASH patients. While 

previous work has suggested an association between the UPR 

and autophagy, our study provides a mechanistic understanding 

of the roles the UPR and CHOP play in regulating saturated 

FFA induced hepatotoxicity at the cellular level. 

Disclosures: 

The following authors have nothing to disclose: Jeffrey A. Willy, Howard C. 

Masuoka, Ronald C. Wek 

321 

Hepatocyte surface localization of rat oatp1a4 requires 

cotrafficking in vesicles that also contain oatp1a1 and 

PDZK1: a new paradigm for subcellular trafficking 

of oatps that do not have a PDZ consensus binding 

domain 

Wen-Jun Wang, John W. Murray, Allan W. Wolkoff; Liver 

Research Center, Albert Einstein College of Medicine, Bronx, NY 

Organic anion transport proteins (oatp) 1a1 and 1a4 mediate 

uptake of endogenous and xenobiotic compounds by hepatocytes. 

In previous studies we showed that oatp1a1 and other 

oatps have a PDZ consensus binding sequence at their C-termini. 

Binding of oatp1a1 to PDZK1 is required for its microtubule-based 

vesicular trafficking to the plasma membrane via 

selective recruitment of specific motor proteins. Rat oatp1a4 

also traffics to the plasma membrane although it, like several 

other oatps, lacks a PDZ consensus binding sequence. In the 

present study, we tested the hypothesis that oatp1a4 traffics 

together with oatp1a1 in common intracellular vesicles, utilizing 

PDZK1 association with oatp1a1 for motor recruitment and 

plasma membrane targeting. Methods: Oatp1a1 and oatp1a4 

were immunolocalized in endocytic vesicles prepared from rat 

as well as wild type and PDZK1 knockout mouse livers. Immunofluorescence 

co-localization of proteins in vesicles was quantified. 

Plasma membrane distribution of oatp1a1 and oatp1a4 

was determined in HEK 293 cells (do not express endogenous 

PDZK1) transiently transfected with oatp1a1, oatp1a4, 

or oatp1a1 plus oatp1a4. Each of these transfections was performed 

with and without cotransfection of PDZK1. Results: 74% 

of vesicles prepared from rat liver that were associated with 

oatp1a4 (n=12,044) were also associated with oatp1a1. 51% 

of oatp1a1 associated vesicles (n=3608) were also associated 

with PDZK1. Similarly, 80% and 64%, respectively, of Oatp1a4-associated 

vesicles that were prepared from wild-type 

and PDZK1 knockout mouse liver were also associated with 

Oatp1a1 (n=5059 and 3415, respectively). Single transfection 

of HEK 293 cells with rat oatp1a1 or oatp1a4 revealed 

predominant cytoplasmic localization with little on the plasma 

membrane. Cotransfection of these cells with PDZK1 revealed 

that oatp1a1 localized to the cell surface while oatp1a4 

remained cytoplasmic. Double transfection with both transporters 

in the absence of PDZK1 revealed that they localized to the 

cytoplasm. In contrast, both oatp1a1 and oatp1a4 localized to 

the cell surface following triple transfection with PDZK1. Conclusions: 

(1) Interaction of rat oatp1a1 with PDZK1 is necessary 

for its trafficking to the cell surface. (2) Although rat oatp1a4 

does not interact with PDZK1, it can co-traffic (“hitchhike”) to 

the cell surface with oatp1a1 by virtue of their colocalization 

in intracellular vesicles containing PDZK1. (3) This provides a 

new paradigm to explain how oatps that lack a PDZ consensus 

binding domain can traffic to the cell surface and suggests 

heterooligomerization of oatps, a mechanism that is currently 

under study. 

Disclosures: 

Allan W. Wolkoff - Consulting: Synageva; Grant/Research Support: Merck 

The following authors have nothing to disclose: Wen-Jun Wang, John W. Murray 

322 

Tissue-Specific Subcellular Localization of Thioesterase 

Superfamily Member 1 (Them1) in the Pathogenesis of 

Nonalcoholic Fatty Liver Disease (NAFLD) 

Yue Li 1 , Susan J. Hagen 2 , David E. Cohen 1 ; 1 Medicine, Brigham 

and Women’s Hospital, Boston, MA; 2 Surgery, Beth Israel Deaconess 


Background: Altered regulation of lipid and glucose homeostasis, 

most often in the setting of insulin resistance and obesity, 

is central to the pathogenesis of NAFLD. Brown adipose tissue 

(BAT) is rich in mitochondria and mediates non-shivering 

thermogenesis in mice. BAT also plays important roles in promoting 

energy expenditure and its suboptimal function may 

contribute to the pathogenesis of obesity in humans. Them1 

is a long chain fatty acyl-CoA thioesterase that is enriched 

in BAT and is upregulated in mice in response to cold stress. 

Them1 -/- mice exhibit marked increases in energy expenditure, 

as well as resistance to diet-induced obesity and NAFLD. 

Under conditions of sustained overnutrition, Them1 appears 

to play a central role in the development of NAFLD both by 

limiting energy expenditure in BAT and by directly promoting 

steatosis in the liver, where it is also upregulated in response 

to high fat feeding. Notwithstanding these striking phenotypes 

and recent reports describing the enzymatic properties of the 

protein, very little is known about the cell biology of Them1. 

Aims: The objective of this study was to elucidate the subcellular 

localization of Them1 and its determinants in BAT and 

liver. Methods: Them1 -/- and wild type mice were subjected 

to cold stress by housing at 4 °C for 24 h or to overnutrition 

by high fat feeding for 16 w. Them1 in liver and BAT was 

visualized by immunofluorescence and confocal microscopy. 

Putative kinases of Them1 phosphorylation sites in BAT identified 

by phosphoproteomics (Cell 2010;143:1174) were predicted 

by sequence analysis (Nat Methods 2014;11:603). 

Them1-EGFP and a deletion mutant were studied using stably 

transfected 3T3-L1 cells, which had been differentiated into 

adipocytes using standard techniques. Results: In BAT of mice


subjected to cold stress, Them1 was strongly localized to lipid 

droplets. By contrast, Them1 in hepatocytes was visualized in 

cytosol, and this was not influenced by high fat feeding. PKCβ 

was the predicted kinase for phosphorylated sites (S15, S18, 

S25) near the N-terminus of Them1. Activation of PKCβ using 

phorbol 12-myristate 13-acetate led to the rapid redistribution 

of Them1-EGFP in a pattern consistent with lipid droplet association. 

The subcellular localization of Them1-EGFP was profoundly 

disrupted by the deletion of a short peptide fragment 

containing the phosphorylation sites. Conclusions: Our data 

demonstrate that cellular localization of Them1 is tissue specific 

and is regulated at least in part by phosphorylation. The tissue 

specific control of Them1 localization may play a key role in 

the pathogenesis of NAFLD. 

Disclosures: 

David E. Cohen - Advisory Committees or Review Panels: Merck, Aegerion, 

Genzyme; Consulting: Intercept 

The following authors have nothing to disclose: Yue Li, Susan J. Hagen 

323 

Longitudinal monitoring of hepatic endoplasmic reticulum 

calcium changes in rats 

Emily Wires, Mark Henderson, Kathleen Trychta, Brandon K. Harvey; 

National Institute on Drug Abuse, Baltimore, MD 

Obesity is a major health concern in developed countries; an 

estimated 1.5 billion individuals are considered overweight 

or obese worldwide. In stressed conditions, such as chronic 

obesity, hepatocytes accumulate excess long chain fatty acids, 

leading to the development of nonalcoholic fatty liver disease 

(NAFLD), representing a spectrum of liver disorders. Moreover, 

obese patients typically exhibit metabolic disorders, cardiovascular 

disease and obesity-related mortality. While the exact 

mechanisms of obesity-associated liver diseases are poorly 

understood, disruptions to endoplasmic reticulum (ER) homeostasis 

have been implicated in disease pathogenesis. The ER 

contains the highest level of intracellular calcium, an estimated 

1,000-10,000 fold greater than cytosolic levels. Perturbations 

to ER calcium levels are associated with a variety of pathologies, 

and have been predicted as a contributing factor to 

the development and progression of obesity-associated disorders. 

Recently, our lab developed a novel secreted ER calcium 

monitoring protein (SERCaMP), to longitudinally monitor ER 

calcium levels in vivo by measuring small volumes of blood; 

a notable enhancement from calcium-specific fluorescent dyes 

that become easily saturated in high levels of calcium and 

are unable to be utilized for in vivo studies. In the present 

study, rats expressing SERCaMP appended to Gaussia luciferase 

(GLuc-SERCaMP) in the liver were fed highly palatable 

foods, colloquially referred to as a cafeteria diet (e.g. cookies, 

pepperoni, potato chips), to mimic dietary intake observed 

in obese individuals. Additional groups included ad libitum 

access to chow and restricted access. Rats exposed to cafeteria 

diet items and ad libitum chow showed significant weight gain 

when compared to restricted rats. Blood samples indicated 

increased levels of circulating GLuc-SERCaMP in both cafeteria 

diet and ad libitum chow, suggesting that a dietary intake, particularly 

over eating, alters ER calcium homeostasis in the liver. 

The half-life of GLuc-SERCaMP is estimated to be less than 10 

minutes; eliminating the possible confound of sensor accumulation 

in the blood. Our study is the first to use a secreted ER calcium 

probe to demonstrate that dietary intake alters ER calcium 

homeostasis in the liver. Furthermore, parallel work has been 

done to monitor hepatic ER stress in order to establish a temporal 

profile between ER calcium dysregualtion and ER stress. 

Disclosures: 

The following authors have nothing to disclose: Emily Wires, Mark Henderson, 

Kathleen Trychta, Brandon K. Harvey 

324 

Mouse CD11b + Kupffer cells recruited from bone marrow 

accelerate liver regeneration after partial hepatectomy 

Hiroyuki Nakashima 1 , Kiyoshi Nishiyama 2 , Masahiro Nakashima 1 , 

Manabu Kinoshita 1 , Shuhji Seki 1 ; 1 Immunology and Microbiology, 

National Defense Medical College, Saitama, Japan; 2 Surgery, 

National Defense Medical College, Saitama, Japan 

Background and Aims: Innate immune cells are known to 

contribute to the liver regeneration after partial hepatectomy 

(PHx). Although TNF and Fas/FasL are the vital components 

for hepatocyte regeneration, the role of Kupffer cells is still in 

controversy. Liver F4/80 + Kupffer cells are classified into two 

subsets; resident radio-resistant CD68 + cells with phagocytic 

and bactericidal activity, and recruited radio-sensitive CD11b +- 

cells with cytokine-producing capacity. The aim of this study is 

to investigate the role of these Kupffer cells in the liver regeneration 

after PHx in mice. Material and Methods: C57BL/6 mice 

were subjected to PHx (70%) or Sham operation. The proportion 

of Kupffer cell subsets in the remnant liver was examined 

by flow cytometry. To examine the role of CD11b + Kupffer cells, 

mice were depleted of these cells before PHx by non-lethal 5 Gy 

irradiation with or without bone marrow transplantation (BMT). 

To investigate the mechanism of macrophage (Mφ) accumulation 

into liver, mice were injected with CCR2 (MCP-1 receptor) 

antagonist and liver regeneration was evaluated. Results: The 

proportion of CD11b + Kupffer cells was greatly increased at 

three days after PHx when the hepatocytes vigorously proliferate, 

whereas the proportion of CD68 + Kupffer cells did not 

significantly change after PHx. Serum TNF levels peaked one 

day after PHx, and intracellular TNF staining revealed that 

CD11b + Kupffer cells are the main source of TNF. Additionally, 

the CD11b + Kupffer cells expressed surface FasL at three days 

after PHx. Serum MCP-1 levels (produced by CD68 + resident 

Kupffer cells) peaked at twelve hours after PHx. Irradiation eliminated 

the CD11b + Kupffer cells for approximately two weeks 

in the liver, while CD68 + Kupffer cells, NK cells and NKT cells 

remained, and hepatocyte regeneration was retarded. However, 

BMT partially restored CD11b + Kupffer cells and recovered 

the liver regeneration. Furthermore, CCR2 antagonist 

treatment decreased the CD11b + Kupffer cells and significantly 

retarded liver regeneration. Conclusion: The CD11b + Kupffer 

cells recruited from bone marrow via the MCP-1/CCR2 and 

TNF and FasL produced by these cells play a pivotal role in the 

liver regeneration after PHx. The possibility is raised that BMT 

can be applied to patients with hepatocellular carcinoma after 

partial hepatectomy for the acceleration of liver regeneration 

and prevention of hepatic failure. 

Disclosures: 

The following authors have nothing to disclose: Hiroyuki Nakashima, Kiyoshi 

Nishiyama, Masahiro Nakashima, Manabu Kinoshita, Shuhji Seki


325 

CD45+ fraction in murine adipose tissue-derived stromal 

cells is therapeutical to Concanavalin A-induced 

murine acute hepatitis model. 

Akihiro Seki 1 , Yoshio Sakai 2 , Masatoshi Yamato 1,2 , Kosuke 

Ishida 1 , Hisashi Takabatake 1 , Alessandro Nasti 1 , Masao Honda 2 , 

Shuichi Kaneko 1,2 ; 1 Disease control and homeostasis, Kanazawa 

university, Kanazawa, Japan; 2 Department of Gastroenterology, 

Kanazawa university hospital, Kanazawa, Japan 

Adipose tissue-derived stromal cells (ADSCs) are expected to 

be useful in regeneration therapy for liver disease because of 

their pluripotency and immunomodulatory capability. However, 

ADSCs are composed of heterogeneous cell fractions, such as 

mesenchymal stem cells, and the regenerative or therapeutic 

effects of all ADSC cell fractions have not been fully elucidated, 

particularly for liver diseases. In this study, we assessed 

whether the substantial ADSC CD45+ leukocyte-lineage fraction 

ameliorates the hepatic inflammatory disease condition 

induced by injecting mice with Concanavalin A (ConA). [Materials 

and Methods] ADSCs were obtained from inguinal subcutaneous 

adipose tissue of C57Bl/6 male mice by collagenase 

digestion. C57Bl/6 female mice were injected intravenously 

with 200 mg ConA to generate the murine hepatitis model. 

The CD45+ fraction, which was about 15% of all ADSCs, was 

isolated with a cell sorter (FACSAriaTMII), and 5 × 10 4 CD45+ 

cells, 1 × 10 6 ADSCs, or PBS were injected intravenously into 

C57BL/6 mice 3 hours after the ConA injection (n = 5 each). 

Sera and liver tissues were obtained from the ConA-injected 

mice after 16 hours to measure serum alanine aminotransferase 

(ALT) and lactate dehydrogenase activities (LDH), and to conduct 

immunohistochemical and gene expression analyses using 

the real-time polymerase chain reaction. CD45+ cells (1 × 10 4 

cells) were also co-cultured with 1 × 10 5 splenocytes stimulated 

with ConA for 2 hours, followed by a gene expression analysis. 

[Results] Injecting 2 × 10 4 CD45+ cells into the ConA-stimulated 

hepatitis mice improved liver congestion compared with 

that of the PBS control. Correspondingly, injecting CD45+ cells 

into ConA-stimulated hepatitis mice suppressed serum ALT and 

LDH activities (P = 0.02 each) but increased hepatic alpha 

fetoprotein and albumin RNA expression (P = 0.01 and 0.02). 

These therapeutic effects of the CD45+ cells injection on the 

ConA-stimulated hepatitis mice were similar to those produced 

after ADSCs were injected. The immunohistochemical analysis 

showed that administrating CD45+ cells ameliorated the 

numbers of CD4+, CD11b+, and Gr-1+ cells infiltrating the 

livers of the ConA-stimulated hepatitis mice. Enhanced interferon-gamma, 

tumor necrosis factor-alpha, and chemokine (C-C 

motif) ligand 3 gene expression by ConA-stimulated splenocytes 

was suppressed when they were co-cultured with CD45+ 

cells in vivo. Thus, the ADSC CD45+ fraction therapeutically 

suppressed ConA-induced mouse hepatitis. [Conclusion] The 

CD45+ leukocyte-lineage fraction in ADSCs, which is uniquely 

immunosuppressive and beneficial, may have important therapeutic 

effects in an acute hepatitis mouse model. 

Disclosures: 






The following authors have nothing to disclose: Akihiro Seki, Yoshio Sakai, 

Masatoshi Yamato, Kosuke Ishida, Hisashi Takabatake, Alessandro Nasti, 

Masao Honda 

326 

Cross-talk between autophagy and the transcription 

factor KLF2 determines endothelial cell phenotype, liver 

microvascular function and hepatic damage after acute 

liver injury. 

Sergi Guixé-Muntet 1 , Fernanda C. de Mesquita 1,3 , Sergi Vila 1 , 

Carmen Peralta 2 , Juan Carlos Garcia-Pagan 1 , Jaime Bosch 1 , Jordi 

Gracia-Sancho 1 ; 1 Barcelona Hepatic Hemodynamic Lab, Hospital 

Clinic de Barcelona - IDIBAPS - CIBEREHD, Barcelona, Spain; 

2 IDIBAPS - CIBEREHD, Barcelona, Spain; 3 Laboratório de Biofísica 

Celular e Inflamação, PUCRS, Porto Alegre-RS, Brazil 

Background & aims: The transcription factor Kruppel-like Factor 

2 (KLF2) can be induced by different drugs, including simvastatin, 

and its expression confers a vasoprotective phenotype to 

the endothelium. Considering recent data suggesting activation 

of autophagy by statins, we aimed at: 1) characterizing the 

molecular relationship between autophagy and KLF2 in the 

endothelium, 2) assessing this relationship in acute liver injury 

due to cold ischemia/warm reperfusion (I/R), and 3) studying 

the effects of modulating KLF2-autophagy in in vitro and ex 

vivo models of acute liver injury. Methods: 1) In vitro: Autophagy 

and KLF2 were modulated and characterized in vascular 

endothelial (HUVEC) and sinusoidal endothelial cells (LSEC) 

cultured in standard conditions or under I/R. KLF2 was induced 

by simvastatin and resveratrol or genetically regulated with 

siRNA or adenovirus in the presence or absence of autophagy 

modulators. Cell viability was evaluated by double-staining 

with acridine orange and propidium iodide, and by trypan 

blue exclusion. Autophagic flux was assessed by western blot 

of LC3B, immunofluorescence of accumulation of autophagosomes, 

and colocalization of autophagosomes & lysosomes. 2) 

Ex vivo: Wistar rats received a) vehicle, b) simvastatin (1mg/ 

kg, i.v.) or c) chloroquine (inhibitor of autophagy; 60mg/kg, 

i.p.) + simvastatin. Livers were explanted, cold stored in Wisconsin 

solution (16h) and warm-reperfused (2h). Microvascular 

function was assessed by liver vasorelaxation to acetylcholine. 

Results: A positive feed-back between autophagy and KLF2 

was observed: KLF2-inducers simvastatin and resveratrol, but 

not the autophagy activator Rapamycin, caused endothelial 

KLF2 overexpression through an autophagy-Rac1-rab7-dependent 

mechanism. In turn, KLF2 induction promoted further 

activation of autophagy. Cold ischemia blunted autophagic 

flux in LSEC. Upon reperfusion, LSEC cold stored in Celsior 

solution showed proper reactivation of autophagy. However, 

LSEC stored in Wisconsin failed to reactivate autophagy, most 

probably due to impairment in autophagosome and lysosome 

fusion. Simvastatin re-activated autophagy by up-regulating 

rab7 (restoring autolysosome formation), which resulted in 

increased KLF2 levels, improved cell viability (in vitro), and 

ameliorated hepatic damage and microvascular function (ex 

vivo). Conclusions: We herein describe for the first time the 

complex link between autophagy and KLF2 modulating the 

phenotype and survival of the endothelium. These results help 

understanding the underlying molecular mechanisms of the 

protection conferred by KLF2-inducers, such as simvastatin, in 

hepatic and extra-hepatic vascular disorders. 

Disclosures: 

Juan Carlos Garcia-Pagan - Consulting: Novartis; Grant/Research Support: 

GORE 

Jaime Bosch - Consulting: Falk, Gilead Science, Intercept Therapeutics, Conatus 

Pharmaceuticals, Exalenz, Almirall, Chiasma 

The following authors have nothing to disclose: Sergi Guixé-Muntet, Fernanda C. 

de Mesquita, Sergi Vila, Carmen Peralta, Jordi Gracia-Sancho


327 

Histone 3 lysine 9 trimethylation (H3K9me3) and 

inflammasome as novel senescent associated markers 

of drug induced premature senescence in hepatocellular 

carcinoma cell lines 

Bijoya Sen 1 , Tarique Anwar 2 , Chhagan Bihari 3 , Archana Rastogi 3 , 

Nirupma Trehanpati 1 , Sukriti Sukriti 1 , Shvetank Sharma 1 , Shiv K. 

Sarin 4 , Gayatri Ramakrishna 1 ; 1 Dept of Research, Institut of Liver 

and Biliary Scienc, Delhi, India; 2 Center for DNA fingerprinting 

and diagnostics, Hyderabad, India; 3 department of pathology, 

institute of liver and biliary sciences, New Delhi, India; 4 institute of 

liver and biliary sciences, New Delhi, India 

Abstract Chemotherapeutic drugs can induce premature 

senescence in cancerous cells. The p53-p21 and pRb-p16 axis 

are the most well studied mechanisms involved in induction 

of senescence. However, these pathways are also associated 

with cell cycle arrest conditions of quiescence, differentiation 

and also cell death. Purpose of this study was to evaluate 

novel markers specific to premature senescence. Hepatocellular 

carcinoma cell lines (Huh7, Hep3B and HepG2) when 

treated with low dose of doxorubicin, a well known chemotherapeutic 

agent, showed permanent cell cycle arrest in G2/M 

phase. Compared to proliferating cells, the senescent cells 

showed enlarged cytomorphology, increased expression of 

cyclin dependent kinase (p21) and positivity for SA-β-galactosidase, 

all indicative of premature senescence. To analyze 

a novel structural marker associated with senescence, transmission 

electron microscopy (TEM) was done which showed 

accumulation of autophagic vacuoles, condensed chromatin 

and prominent mitochondrial-endoplasmic reticulum (ER) 

coupling which were conspicuosly absent in non-senescent 

hepatocytes. The changes in chromatin was further evaluated 

by immunofluorescence microscopy which showed presence of 

heterochromatic foci only in the senescent cells. Hetrochromatin 

formation in turn involves histone modification and senescent 

cells showed a significant increase in inactive chromatin mark 

viz., H3K9 trimethylation compared to non-senescent cells. Further, 

microarray based gene expression analysis was done to 

identify novel pathways uniquely associated with senescence 

which showed a differential increase in cytokine-chemokine 

response. This in turn was supported by the fact that the secretome 

of senescent cell was highly effective in closure of wound 

as assessed by the wound healing assay, unlike the control 

cell secretome. The active senescent secretome was associated 

with increased transcript level of PyCard/Asc, and IL-1β indicative 

of inflammasome activation. In conclusion we have now 

identified inflammasomes and H3K9 trimethylation as specific 

senescence associated markers. This project was funded by 

Dept of Biotechnology, India. 

328 

Netrin-1 Promotes Liver Cancer Cells Collective Invasion 

in a 3D Cell Culture Model 

Ping Han 1 , Yu Fu 2 , Jingmei Liu 1 , Dongxiao Li 1 , Yunwu Wang 1 , 

Dean Tian 1 , Wei Yan 1 ; 1 Gastroenterology, Tongji Hospital, Tongji 

Medical College, Huazhong University of Science and Technology,Wuhan,China, 

Wuhan, China; 2 Gastroenterology, Union Hospital, 

Tongji Medical College, Huazhong University of Science and 

Technology,Wuhan,China, Wuhan, China 

Collective invasion is the new understanding of tumor metastasis, 

while the mechanism needs to be further studied. Our 

previous studies showed that Netrin-1 promoted the invasion 

ability of liver cancer cells(LCCs). Immunohistochemical studies 

showed that LCCs invaded surrounding tissue as collective 

invasion. The expression of Netrin-1 in these LCCs was 

increased. In this study, we performed a 3D cell culture model 

to further confirm the role of Netrin-1 in LCCs collective invasion. 

We designed a 3D cell culture model, after centrifuge, 

5×10 6 cells were resuspended by 120μl 10% sucrose solutions, 

and quickly mixed with equal volume of 0.5% hydrogel 

solution( BeaverNano ), the cells were then seeded in a Class 

Bottom Cell Culture Dish(NEST), and allowed to grow for six 

days. Then, cells were observed under a confocal microscopy. 

The number and distance of collective invasion in LCCs were 

evaluated after enhanced or silenced expression of Netrin-1. 

The N-cadherin shRNAs were used to verify the role of N-cadherin-based 

junctions in Netrin-1 promoted collective invasion. 

We found the number and distance of invaded SK-Hep1 cells, 

a high metastasis potential cell line, were significantly more 

than Huh7 cells with low metastasis potential, the normal liver 

cell line LO2 cells nearly had no cell invaded. Meanwhile, 

the invaded cells had significant more expression of Netrin-1. 

Overexpression of Netrin-1 increased collective invasion in the 

3D cell culture model and elevated N-cadherin expression in 

Huh7 cells, while stable knockdown of Netrin-1 remarkably 

inhibited collective invasion and decreased N-cadherin expression 

in SK-Hep1 cells. Interestingly, knockdown N-cadherin in 

Huh7 cells significantly diminished Netrin-1 promoted liver cancer 

cell collective invasion. These results suggest that Netrin-1 

enhances N-cadherin junctions to promote LCCs collective invasion, 

subsequently may increase LCCs metastasis.(This study 

is supported by the National Natural Science Foundation of 

China (81472311), the Fundamental Research Funds for the 

Central Universities (2014ZHYX020,2014TS077), the Project 

sponsored by SRF for ROCS, SEM (2014-1685) and Hubei 

Province health and family planning scientic research project 

(WJ2015Q006)) 

Collective invasion of SK-Hep1 cells in 3D cell culture 

Disclosures: 

The following authors have nothing to disclose: Ping Han, Yu Fu, Jingmei Liu, 

Dongxiao Li, Yunwu Wang, Dean Tian, Wei Yan 

Disclosures: 

The following authors have nothing to disclose: Bijoya Sen, Tarique Anwar, 

Chhagan Bihari, Archana Rastogi, Nirupma Trehanpati, Sukriti Sukriti, Shvetank 

Sharma, Shiv K. Sarin, Gayatri Ramakrishna


329 

Interaction with collagen matrix modulates macrophage 

expression of MMPs in part by controlling cell shape. 

Toshihiko Matsumoto 1 , Stephen Jenkins 3 , Koichi Fujisawa 2 , Taro 

Takami 2 , Naoki Yamamoto 2 , John P. Iredale 3 , Isao Sakaida 2 ; 

1 Department of Gastroenterology & Hepatology, Department of 

Oncology and Laboratory Medicine, Yamaguchi University Graduate 

School of Medicine, Ube, Japan; 2 Department of Gastroenterology 

& Hepatology, Yamaguchi University Graduate School of 

Medicine, Ube, Japan; 3 The Queen’s Medical Research Institute, 

University of Edinburgh, Edinburgh, United Kingdom 

Background and Objectives: Macrophages have a pivotal 

role in resolution of liver fibrosis, during which, pro-fibrotic 

macrophages undergo a switch in phenotype and up-regulate 

expression of MMP9, MMP12 and MMP13, which promote 

myofibroblast apoptosis and degradation of the scar extracellular 

matrix (ECM). However, the signals that regulate this balance 

of opposing functions remain to be fully determined. It was 

recently reported that cell shape had an important role in regulating 

macrophage function. We hypothesized that alterations 

in cell shape associated with changes in ECM architecture may 

provide cues to modulate the phenotype and MMPs expression 

of macrophages in fibrotic liver. We therefore examined the 

effects of collagen on macrophage shape and MMP expression 

in vitro, and investigated the underlying mechanisms linking 

these processes with the aim of developing future anti-fibrotic 

therapies. Method: Mouse bone marrow derived macrophages 

were treated with GM-CSF (GM-BMDM) to model inflammatory 

macrophages and subsequently cultured on a monolayer of 

collagen I, plastic alone or BSA control and the shape, size 

and MMP expression determined. To clarify the relationship 

between effects on cell shape and MMPs expression, actin 

polymerization of GM-BMDM cultured on Collagen I or plastic 

was assessed by phalloidin staining. GM-BMDM were then 

treated with Cytochalasin D and Rho family GTPase inhibitors, 

such as NSC23766 and ML141, to inhibit actin polymerization 

and determine the effect on MMP expression. Result: Culture 

on collagen I inhibited expression of MMP9, 12 and 13 

by GM-BMDM (p


331 

Genetic Induction of Metabolically Functional, Polarized 

Cultures of Proliferating Human Hepatocytes 

Gahl Levy 1 , Stefan Heinz 3 , Ella H. Sklan 2 , Oren Shibolet 2 , Joris 

Braspenning 3 , Yaakov Nahmias 1 ; 1 Bioengineering, The Hebrew 

University of Jerusalem, Jerusalem, Israel; 2 Tel Aviv University, Tel 

Aviv, Israel; 3 Medicyte, Heidelberg, Germany 

Primary hepatocytes are responsible for the modification, clearance, 

and transformational toxicity of most xenobiotics. Regretfully, 

like other primary cells, hepatocytes do not proliferate 

in vitro, while their transformation causes loss of metabolic 

function. Here, we report a method to conditionally induce 

proliferation in primary human hepatocytes by genetic modification 

(upcyte ® process). Proliferating human hepatocytes can 

undergo up to 40 population doublings, producing 10 13 to 

10 16 cells, and differentiate when contact-inhibited into metabolically 

functional, polarized cells with functional bile canaliculi. 

The cells lack fetal and tumor markers and show high levels 

of CYP3A4 expression and function. Differentiated cells show 

gene expression, CYP450 activity, induction, and TC 50 

toxicity 

profile comparable to primary human hepatocytes. Importantly, 

proliferating hepatocytes can be readily infected with Hepatitis 

C Virus (HCV) showing replication and infectivity profile comparable 

to Huh7.5.1 cells. Our results offer a promising new 

technology to expand human hepatocytes from varying genetic 

backgrounds for scientific research, clinical applications, and 

pharmaceutical development. 

Disclosures: 

Oren Shibolet - Advisory Committees or Review Panels: MSD, Abbvie, Novartis; 

Grant/Research Support: MSD 

The following authors have nothing to disclose: Gahl Levy, Stefan Heinz, Ella H. 

Sklan, Joris Braspenning, Yaakov Nahmias 

332 

Hepatocyte Isolation from Liver Fine Needle Aspiration 

and Core Needle Biopsy 

Andrew H. Talal 1,2 , Rosemary Furlage 3 , Yvonne J. Woolwine-Cunningham 

1 , Jun Li 4 , Robin Difrancesco 4,5 , Gene D. Morse 4,5 , Georg 

Lauer 6 , Jun Qu 4 , Paul K. Wallace 3 ; 1 Center for Clinical Care and 

Research in Liver Disease, SUNY-Buffalo, Buffalo, NY; 2 Division of 

Gastroenterology, Hepatology and Nutrition, Department of Medicine, 

SUNY-Buffalo, Buffalo, NY; 3 Department of Flow & Image 

Cytometry, Roswell Park Cancer Institute, Buffalo, NY; 4 School of 

Pharmacy and Pharmaceutical Sciences, SUNY-Buffalo, Buffalo, 

NY; 5 NYS Center of Excellence in Bioinformatics and Life Sciences, 

University at Buffalo, Buffalo, NY; 6 Gastrointestinal Unit, Massachusetts 

General Hospital, Boston, MA 

Background: Although the liver is the major site of drug metabolism, 

little is known about the determinants of intrahepatic drug 

concentration. We sought to develop techniques for hepatocyte 

isolation for measurement of intrahepatic drug concentration. 

Methods: Thirteen Sprague Dawley rats had liver specimens 

collected via FNA (21G needle), core needle biopsy (CNB) 

(16G needle), and surgical resection. Blood was collected by 

cardiac puncture at the same time. The optimized procedure 

initially requires the addition of chilled washing solution to 

the sample and centrifugation at 150G for 3 min at 4 o C. Subsequently, 

the sample is incubated with collagenase at 37 o C 

while rocking for 30 minutes, pushed through a 100m filter, 

centrifuged and washed twice more. A sample aliquot is subsequently 

counted (Countess, Invitrogen) and analyzed (Fortessa, 

Becton Dickinson). On one animal, proteins were initially 

extracted with a Polytron sonication procedure, separated using 

a nano-LC system coupled to an Orbitrap Fusion Mass Spect on 

a 100-cm-long column. Results: Average length and weight of 

the CNB samples was 2.0 cm and 10 mg, respectively, while 

FNA samples ranged from 3-12 mg. The total number of cells 

isolated, the percentage of hepatocytes in the sample, and the 

number of sorted hepatocytes is shown (Table). Combining 

three FNA samples together increased cell yield to 222,308 

with 46% hepatocytes resulting in 89,358 sorted hepatocytes. 

Combining five FNA samples together resulted in a very similar 

number of sorted hepatocytes. Samples weighing 8.0 and 

20.6 mgs yielding 427.2 and 2670.6 mg, respectively, were 

used for proteomic analysis. A total of 879 and 1299 proteins 

were isolated from the FNA and CNB samples, respectively, of 

which those involved in oxidative reduction and responses to 

organic substances had the highest percentages (FNA: 34%, 

CNB: 28%). Mitochondrion and cytosol protein analytes were 

the most frequently identified cellular components (FNA: 60%; 

CNB: 52%). Conclusions: One FNA pass results in sufficient 

hepatocyte yield that should be suitable for measurement of 

intrahepatic drug concentration and for proteomic analysis. 

Combining three FNA passes into the same sample increases 

the cell yield proportionally. Isolation of nonparenchymal cells 

from the same sample should also be feasible. 

Cell Yields 

SD, standard deviation; Hep, hepatocytes 

Disclosures: 

Andrew H. Talal - Advisory Committees or Review Panels: Merck and Co, Abbvie, 

Gilead; Board Membership: Pfizer; Grant/Research Support: Merck and Co, 

Gilead, Abbott Molecular, Abbvie 

The following authors have nothing to disclose: Rosemary Furlage, Yvonne J. 

Woolwine-Cunningham, Jun Li, Robin Difrancesco, Gene D. Morse, Georg Lauer, 

Jun Qu, Paul K. Wallace 

333 

Characterization of Microtubule Motor Proteins in the 

Endocytic and Autophagic Compartments During Nutritional 

Stress and Aging 

Eloy Bejarano-Fernandez, John W. Murray, Xintao Wang, Ana 

Maria Cuervo, Allan W. Wolkoff; Albert Einstein College of Medicine, 

Bronx, NY 

Purpose: These studies were designed to survey and functionally 

analyze microtubule motor proteins present on autophagic 

and endocytic organelles from liver in response to nutritional 

stress and aging. Methods: Organelles were purified from livers 

of 3-month and 22-month old mice; motor proteins and 

organelle markers were analyzed by western blot. Primary 

fibroblasts generated from the same animals were subject to fluorescent 

microscope imaging and automated analyses. Results: 

A detailed analysis of motor proteins present on intracellular 

organelles revealed a high association of dynein with autophagosomes 

(Enrichment Versus Homogenate (EVH): 2.39 ± 0.28) 

compared to endosomes (EVH 1.01 ± 0.37), and lysosomes 

(EVH 0.60 ± 0.06). Kif5b (kinesin-1) showed more equal distribution 

between organelles (EVH: 0.57 ± 0.30, 0.27 ± 0.04, 

0.22 ± 0.09 for endosomes, autophagosomes and lysosomes, 

respectively). Kif3A (kinesin-2) was abundant on endosomes 

(EVH: 1.61 ± 1.57) and autophagosomes (EVH: 0.56 ± 0.15) 

but not lysosomes (EVH: 0.14 ± 0.03). Since dynein was relatively 

low in liver lysosomes, minus end directed kinesins were 

examined. Among these, KifC3 was strongly enriched in lysosomes 

(EVH: 12.08 ± 4.74) as compared to autophagosomes 

(EVH: 0.56 ± 0.35) and endosomes (EVH: 1.12 ± 0.50). Aged 

mice were found to exhibit an 84.5% decrease in the level


of dynein in autophagosomes (p=0.002). Interestingly, the 

age-dependent differential recruitment is lost under acute starvation. 

A similar behavior was also found for the KifC3 motor 

in lysosomes. Live cell vesicle tracking revealed differences in 

intracellular motility that correlated with these findings. The 

motility of autophagosomes was reduced by 16% in cells from 

aged mice (p=0.03, p>0.05). In addition, a dynein inhibitor 

(EHNA) inhibited autophagosome motility (-24.6%, p=0.02) 

exclusively in young mice. Upon serum starvation, no differences 

were found in autophagosomal motility pointing out the 

recruitment of motors during nutrient deprivation. In agreement 

with these results, in vitro assays showed that on average 22% 

of the purified organelles moved along microtubules whereas 

30% of autophagolysosomes from starved mice were motile 

(p=0.04). Conclusions: Our analysis provides for first time 

quantitative biochemical and imaging evidence for the impact 

of aging in the association of motors to degradative compartments 

under basal and nutritional stress-induced autophagy. 

Disclosures: 


The following authors have nothing to disclose: Eloy Bejarano-Fernandez, John 

W. Murray, Xintao Wang, Ana Maria Cuervo 

334 

Acute alcohol binge causes skeletal muscle mitochondrial 

intermediate metabolite deficiency 

Allawy Allawy 2,3 , Gangarao Davuluri 2 , Avinash Kumar 2 , Bin Gao 4 , 

Ming-Jiang Xu 4 , Megan R. McMullen 2 , Rebecca L. McCullough 2 , 

Srinivasan Dasarathy 1,2 ; 1 Department Of Gastroenterology and 

Hepatology, Cleveland Clinic, Cleveland, OH; 2 Department Of 

Pathobiology, Cleveland Clinic, Cleveland, OH; 3 Department Of 

Internal medicine, Cleveland Clinic, Cleveland, OH; 4 Laboratory 

of Liver Diseases. NIAAA, NIH, Bethesda, MD 

Introduction Binge drinking is believed to result in more severe 

tissue injury than chronic ethanol exposure. There are no data 

on the comparing binge drinking and chronic ethanol exposure 

on skeletal muscle mitochondrial function. We used multiple 

models of in vivo and an in vitro model of skeletal muscle 

ethanol exposure to determine its impact on tricarboxylic acid 

cycle intermediates. Materials and Methods. Murine C2C12 

myotubes were exposed to 100mM ethanol for 6 h and 24 h 

and lysates used to quantify TCA cycle intermediates. In vivo 

studies were performed in mice exposed to different patterns of 

ethanol exposure and included chronic alcohol feeding for 25 

or 56 days, ethanol feeding for 10 days followed by a single 

binge, twice weekly ethanol feeding for 8 weeks, pair fed 

or maltose fed controls. Gastrocnemius muscle was harvested 

and TCA cycle intermediates extracted using ethyl acetate, 

tertbutyldimethylsilyl derivatives generated and quantified by 

gas chromatography-mass spectrometry using protocols established 

in our laboratory. All experiments were performed in 

3-4 mice in each group and 6 independent experiments in 

myotubes. All data are expressed as mean±SD of percentage 

change after normalization against the concentrations in the 

appropriate controls. Results. In the chronic ethanol exposure 

for either 25 or 56 days, no significant changes were observed 

in the skeletal muscle concentration of citrate, αketoglutarate, 

succinate, fumarate or malate. Interestingly, compared to pair 

fed controls (100%), in the single binge model, concentration 

of citrate was 49.7±8.9% (p


hepatic injury, increase macrophage recruitment, and increase 

hepatic expression of CXCL1 following IRI. The dramatic reduction 

in tissue injury by MSC-EV support the therapeutic use of 

MSC-EV to reduce hepatic IRI following hepatic surgery or in 

hepatic transplantation. 

Disclosures: 

The following authors have nothing to disclose: Hiroaki Haga, David D. Lee, 

Sarah Nix, Irene K. Yan, Tushar Patel 

336 

Liver Fabrication Using Decellularized Whole-Organ 

Scaffold for Transplantable Graft in large animal model 

Hiroshi Yagi, Kenta Inomata, kazuki tajima, Taizo Hibi, Yuta Abe, 

Minoru Kitago, Masahiro Shinoda, Osamu Itano, Yuko Kitagawa; 

Surgery, Keio University School of Medicine, Shinjuku-ku, Japan 

Background: Whole-organ scaffold generated by tissue decellularization 

technology can be one of the solutions to fabricate 

transplantable organ graft, which contains functional cell 

types including progenitor cells. However, the feasibility of this 

technology for human-scale application, and the biological 

alterations of the scaffold after implantation are still unclear. 

Methods: To determine if the decellularization methodology 

could be applied in large animal model and it could be sufficiently 

recellularized with primary cells as well as human iPS 

cell derived hepatic progenitor cells, the decellularized scaffold 

was first generated using SDS, TrytonX-100 and CHAPS. 

Next, endothelial cells and primary porcine hepatocytes were 

infused into the scaffold under the monitoring of intravascular 

pressure. Finally, the fabricated liver was implanted into the 

porcine body and evaluated by angiography after vascular 

anastomoses. Histological study was performed to evaluate cell 

infiltration, degree of coagulation and adhesion of the scaffold. 

In addition, the liver scaffold containing human iPS cell derived 

liver progenitor cells was transplanted into porcine body under 

immune-suppression. Results: The endothelial cells could cover 

most of the vessel lumen of the scaffold and the primary hepatocytes 

were well distributed into the parenchymal space. The 

fabricated liver could be successfully transplanted into porcine 

body connecting with portal vein and IVC. The graft was well 

perfused and preserved in the porcine abdominal cavity without 

bleeding or absorption. Interestingly, histological analysis 

of transplanted scaffold with human iPS derived progenitor 

cells revealed liver specific architecture with ALB positive cells 

in parenchymal space. Conclusions: We could scale-up and 

optimize the liver fabrication system to apply not only primary 

cells but also human iPS derived cells into clinically feasible 

model. 

Disclosures: 

The following authors have nothing to disclose: Hiroshi Yagi, Kenta Inomata, 

kazuki tajima, Taizo Hibi, Yuta Abe, Minoru Kitago, Masahiro Shinoda, Osamu 

Itano, Yuko Kitagawa 

337 

Three-dimensional bioactive human liver acellular scaffold 

with preserved architecture and biomechanical 

properties 

Giuseppe Mazza 1 , Walid Al-Akkad 1 , Lisa Longato 1 , Andrea 

Telese 1 , Andrew R. Hall 1 , Luca Urbani 2 , Benjamin Robinson 3 , 

Giusi Marrone 1 , Oliver Willacy 1 , Luca Frenguelli 1 , Marco Curti 1 , 

Massimo Malago 1 , Tu Vinh Luong 1 , Kevin Moore 1 , Armando E. 

Del Rio Hernandez 3 , Paolo De Coppi 2 , Krista Rombouts 1 , Massimo 

Pinzani 1 ; 1 Institute for Liver and Digestive Helath, UCL, London, 

United Kingdom; 2 Institute for Child Health, UCL, London, United 

Kingdom; 3 Imperial College, London, United Kingdom 

Background: Human tissue engineering combines cells and 

extracellular matrix (ECM) saffolds for the development 

of 3D-structure in order to regenerate tissues/organs and 

to recapitulate disease in vitro. The key challenge in tissue 

engineering is the development of biomaterials that mimic 

the complexity of human tissue. Techniques for tissue decellularization 

have been introduced in order to obtain natural 

acellular scaffold. Optimal scaffolds should be characterised 

by preserved ECM integrity, bioactivity and three-dimensional 

organisation. Aim: The aim of this study was to develop a rapid 

protocol for the decellularisation of small samples of healthy 

human liver and demonstrate preservation of ECM composition, 

3D-architecture, bioactivity and biomechanical properties. 

In addition, repopulation with cultured human liver cell lines, 

namely hepatoblastoma (HepG2), hepatic stellate (LX2) cells, 

and human umbilical vein endothelial cells (HUVEC). Methods: 

Decellularisation of healthy human liver cubes (i.e. 125mm 3 ) 

was completed within 3 hours of agitation in solutions with 

detergents and enzymes. The decellularisation efficiency was 

determined by immunohistochemistry for ECM components and 

residual DNA, scanning electron and second harmonic generation 

microscopy, proteomic, raman spectroscopy, atomic force 

microscopy and chorioallantoic membrane assay. RESULTS: 

This innovative protocol resulted in liver cube scaffolds with 

a preserved 3D structure and ECM composition, while DNA 

and cellular residues were successfully removed. Tissue stiffness 

was preserved in the decellularized tissues (2KPa) and 

the acellular liver maintained the capability to induce neo-angiogenesis 

after 7 days. Interestingly, HUVEC repopulated the 

decellularised vessels within the liver scaffold expressing functional 

marker such as FVIII. Acellular human liver repopulated 

with LX2 and HepG2 cells showed remarkable difference in 

gene and protein expression when compared with 2D-system. 

COL1A1 gene was less expressed in LX2 cells when cultured 

in 3D while TGFB1 and LOX were increased. Notably, albumin 

mRNA expression was strongly upregulated in HepG2 

cells after 14 days in 3D-culture. CONCLUSION: This is the first 

report describing a protocol for fast human liver tissue cubes 

decellularization. The decellularization protocol maintained the 

natural 3D structure and ECM composition and organisation of 

human liver tissue. This is a key advance in the development 

of 3D technologies for the study of liver disease as well as for 

the development of 3D-carrier for hepatocyte transplantation. 

Disclosures: 

Kevin Moore - Advisory Committees or Review Panels: Servier 

Massimo Pinzani - Advisory Committees or Review Panels: Intercept Pharmaceutical, 

Silence Therapeutic, Abbot; Consulting: UCB; Speaking and Teaching: 

Gilead, BMS


The following authors have nothing to disclose: Giuseppe Mazza, Walid Al-Akkad, 

Lisa Longato, Andrea Telese, Andrew R. Hall, Luca Urbani, Benjamin Robinson, 

Giusi Marrone, Oliver Willacy, Luca Frenguelli, Marco Curti, Massimo 

Malago, Tu Vinh Luong, Armando E. Del Rio Hernandez, Paolo De Coppi, Krista 

Rombouts 

338 

Promotion of liver transplant tolerance by hepatic plasmacytoid 

dendritic cells 

Osamu Yoshida 2,1 , Shoko Kimura 2 , Benjamin M. Matta 2 , Yusuke 

Imai 1 , Masanori Abe 1 , Yoichi Hiasa 1 , Angus W. Thomson 2,3 ; 

1 Department of Gastroenterology and Metabology, Ehime University 

Graduate School of Medicine, Ehime, Japan; 2 Starzl 

Translantation Institute, University of Pittsburgh School of Medicine, 

Pittsburgh, PA; 3 Department of Immunology, University of 

Pittsburgh School of Medicine, Pittsburgh, PA 

Background: Plasmacytoid dendritic cells (pDC) are an 

important source of interferon-alpha and serve as a first line 

of defense against viral infections. On the other hand, pDC, 

especially liver pDC, display tolerogenic properties and play 

an important role in induction of oral tolerance. The liver is 

both an immune and tolerogenic organ that is accepted without 

immunosuppressive therapy when transplanted across MHC 

barriers in mice. We have reported previously that myeloid 

DC can promote the induction of transplant tolerance in mouse 

models. Here we hypothesized that liver pDC might play an 

important role in immune regulation and the promotion of liver 

transplant tolerance. Methods: Liver pDC were depleted from 

10-12 week-old male C57BL/6 (B6; H2 b ) mice by intravenous 

injection of mPDCA (120G8) mAb. Profound pDC depletion 

was confirmed by flow cytometry. Untreated or pDC-depleted 

B6 livers were transplanted orthotopically into normal 10-12 

week-old male C3H (H2 k ) mice. Graft rejection was assessed 

by recipient survival and serum ALT levels and histology on day 

4 after transplantation. We also assessed the impact of pDC 

on alloimmunity using a delayed-type hypersensitivity model. 

Normal C3H mice were sensitized with normal or pDC-depleted 

donor (B6) splenocytes (10 7 , sc) 2 weeks before ex 

vivo re-stimulation of host T cells with donor antigen. T cell 

proliferation and cytokine production were assessed by CFSE- 

MLR and ELISA, respectively. Results: More than 90% of pDC 

were depleted in both liver and spleen. Whereas mice transplanted 

with normal livers (n= 6) survived >100 days without 

any immunosuppression, 83% (n=5/6) mice that received 

pDC-depleted livers died within 55 days (MST: untreated: 

>100d vs pDC-depleted 25d; p


340 

Novel Vasodilator and Anti-inflammatory Drug-based 

Strategy for Restoring Ischemia/ reperfusion-Exposed 

Donor Liver to Healthy State 

Thirunavukkarasu Chinnasamy 1,2 , Fadi L. Jaber 1 , Sanjeev Gupta 3 ; 

1 Department of Medicine, Albert Einstein College of Medicine, 

Bronx, NY, USA, Bronx, NY; 2 Department of Biochemistry and 

Molecular Biology, Pondicherry University, Puducherry, India; 

3 Departments of Medicine and Pathology, Marion Bessin Liver 

Research Center, Diabetes Center, Cancer Center, Ruth L. and 

David S. Gottesman Institute for Stem Cell and Regenerative Medicine 

Research, Albert Einstein College of Medicine, Bronx, NY 

Maintaining donor organs in healthy state is critical for isolating 

viable cells and for liver transplantation. A major cause of 

donor organ deterioration is ischemia/reperfusion (IR) injury 

due to complex events, e.g., vasoconstriction or inflammation 

with release of ROS, cytokines/chemokines/receptors, 

etc. We hypothesized that donor organ preconditioning with 

early control of vasoconstriction and inflammation by NO and 

TNF-α blockade as master regulators of interest will be effective. 

Therefore, we used NO donor, nitroglycerin (NG), and 

TNF-α blocker, Etanercept (ETN), before nonlethal hepatic IR in 

F344 rats with clamping of portal and hepatic blood flow for 

15 min and reperfusion for 24h. In drug-untreated controls, IR 

produced much oxidative stress, including hepatic GGT expression, 

nitrotyrosine, 8-oxo-dG DNA adducts and liver necrosis, 

as expected. In hepatocytes isolated from donors with IR, cell 

viability and attachment in dishes was inferior to control donor 

cells without IR. However, cultured cells from IR donors better 

withstood secondary H 2 

O 2 

or CCl 4 

injury. By contrast, preconditioning 

with ETN 12-18h before I/R and/or NG starting 

5min before IR and continuing during and after IR for 20 min 

lessened hepatic injury. ETN+NG combination was most effective 

versus either drug alone in restoring IR liver to near-normal 

without necrosis or GGT and nitrotyrosine expression. Hepatocytes 

from ETN+NG-preconditioned rats with IR were more 

viable, attached better in dishes, and were also more sensitive 

to H 2 

O 2 

and CCl 4 

injury, which was similar to healthy control 

hepatocytes. FACS analysis showed that polyploid hepatocytes 

were depleted in cells from IR rats but not in cells from healthy 

control and ETN+NG preconditioned donors, again indicating 

less oxidative insult. Next, to further examine potential of 

isolated donor cells, we performed cell transplantation assays 

in DPPIV- rats. Hepatocytes from healthy donor rats engrafted 

and were normally distributed in periportal areas in zones 

1/2 of liver lobules. However, hepatocytes from IR donor rats 

engrafted less and were mainly in portal vein radicles, suggesting 

differences in their cell adhesion and matrix-interacting 

properties. By contrast, engraftment and intrahepatic distribution 

of transplanted hepatocytes isolated from ETN+NG preconditioned 

livers was similar to cells from healthy control rats. 

Conclusions: The donor liver was successfully preconditioned 

and restored to normal health with a simple regimen of NO-donor 

and TNF-α blockade. This drug approach will be readily 

translated to clinical setting for improving outcomes in liver 

transplantation and use of donor livers for cell therapy. 

Disclosures: 

The following authors have nothing to disclose: Thirunavukkarasu Chinnasamy, 

Fadi L. Jaber, Sanjeev Gupta 

341 

Identification of microRNAs Associated with Allograft 

Tolerance 

Matthew J. Vitalone 1 , Liang Wei 2 , Karine Piard-Ruster 1 , Carlos 

O. Esquivel 1 , Olivia M. Martinez 1 , Sheri M. Krams 1 ; 1 Surgery, 

Stanford University School of Medicine, Stanford, CA; 2 Sichuan 

Academy of Medical Sciences and Sichuan Provincial People’s 

Hospital, Chengdu, China 

Introduction: Although the liver is less immunogenic than 

other solid organs, liver transplant recipients receive lifelong 

immunosuppression. The aim of this study was to profile the 

expression of microRNAs (miRNAs) associated with the induction 

and maintenance of tolerance to an allograft. Methods: 

Previous studies showed that donor-specific tolerance can be 

induced in recipients of rat orthotopic liver transplants (OLT) 

after post-transplant total lymphoid irradiation (TLI). To identify 

miRNAs associated with tolerance we profiled liver grafts from 

syngeneic (DAàDA) and allogeneic OLT recipients (DAàLewis) 

that received post-transplant TLI, allograft recipients that were 

not treated post-transplant, and normal DA livers. Untreated 

allograft recipients reject their grafts within 10 days whereas 

TLI treated recipients have long-term graft survival (>100 days), 

thus miRNAs were examined at two time points, seven days 

(induction of tolerance) and 100 days (established tolerance) 

post-transplant Results: A supervised principal component analysis 

(PCA) using the top 15 differentially altered miRNA was 

able to robustly model the study groups with as little as three 

principal components describing 88.5% of the variance of 

the data. Indeed, three miRNA, miR142-3p, miR142-5p and 

miR181a were determined to be key miRNA associated with 

tolerance. Further, analysis of the full complement of miRNAs, 

by PCA, revealed that both the established tolerance and the 

syngeneic groups occupied the same relative three-dimensional 

space close to the normal liver group, indicating similar miRNA 

profiles. Conclusions: A small group of miRNA can distinguish 

graft status post-transplant. Further, our findings support a 

model whereby tolerant liver allografts express a profile consistent 

with that of healthy livers. 

Disclosures: 

The following authors have nothing to disclose: Matthew J. Vitalone, Liang Wei, 

Karine Piard-Ruster, Carlos O. Esquivel, Olivia M. Martinez, Sheri M. Krams 

342 

Previous ischemic preconditioning in experimental 

model of steatotic liver trasplantation with brain death 

Mónica B. Jiménez-Castro 1 , Mariana Mendes-Braz 1 , Jordi 

Gracia-Sancho 3 , Maria Eugenia Cornide-Petronio 1 , Araní 

Casillas-Ramírez 2 , Juan Rodes 1 , Carmen Peralta 1 ; 1 Institut D’Investigacions 

Biomèdiques August Pi i Sunyer, Barcelona, Spain; 2 Hospital 

Regional de Alta Especialidad de Ciudad Victoria, Ciudad 

Victoria, Mexico; 3 Barcelona Hepatic Hemodynamic Laboratory, 

IDIBAPS, CIBEREHD, Institut D’Investigacions Biomèdiques August 

Pi i Sunyer, Barcelona, Spain 

Background & Aims: Liver transplantation has evolved to 

become a standard therapy for certain end-stage liver diseases. 

Nowadays, high percent of organs come from donors who 

have suffered brain trauma-brain dead donors, which may also 

show hepatic steatosis, being both characteristics risk factors 

in liver transplantation. Nevertheless brain dead reduces the 

tolerance of liver grafts to the preservation/reperfusion injury 

and reduces graft survival. Ischemic preconditioning shows 

benefits when applied in the liver transplantation from nonbrain 

dead patients like hepatectomies, whereas it has been 

less promising in the transplantation from brain dead patients.


This study examined how brain death affects preconditioned 

steatotic liver grafts undergoing transplantation. Methods: Steatotic 

grafts from non-brain dead and brain dead-donors were 

cold stored for 6 hours and then transplanted. After 4 hours of 

reperfusion, hepatic damage was analyzed. Two strategies, 

acetylcholine pre-treatment and ischemic preconditioning or 

the combination of them were evaluated and their underlying 

mechanisms were characterized. Results: The presence 

of brain dead exacerbated hepatic damage in steatotic liver 

since increased transaminase levels and damage score in 

comparison with the liver transplantation group. Acetylcholine 

treatment reduced hepatic damage after liver transplantation 

in brain dead donors, through PKC, increased antioxidants 

and reduced lipid peroxidation, nitrotyrosines and neutrophil 

accumulation,. On the other hand, preconditioning benefits in 

non-brain dead donors were associated with nitric oxide and 

acetylcholine generation. In brain dead donors, preconditioning 

generated nitric oxide but did not promote acetylcholine 

up-regulation, and this resulted in inflammation and damage. 

Conclusions: We suggest that the combination of acetylcholine 

treatment and preconditioning conferred stronger protection 

against damage, oxidative stress and neutrophil accumulation 

than acetylcholine treatment alone. These superior beneficial 

effects were due to a selective preconditioning-mediated 

generation of nitric oxide and regulation of PPAR and TLR4 

pathways, which were not observed when acetylcholine was 

administered alone. We herein propose that the time frame 

between the declaration of brain dead and organ retrieval 

provides an important window for cytoprotective intervention, 

which may counteract the detrimental effects of brain dead. 

Consequently, our findings show that the combination of acetylcholine 

and preconditioning as a feasible and a protective 

strategy to reduce the adverse effects of brain dead and to 

improve the quality of liver grafts. 

Disclosures: 

The following authors have nothing to disclose: Mónica B. Jiménez-Castro, Mariana 

Mendes-Braz, Jordi Gracia-Sancho, Maria Eugenia Cornide-Petronio, Araní 

Casillas-Ramírez, Juan Rodes, Carmen Peralta 

343 

WITHDRAWN 

344 

Survival After Intra-Arterial Treatment for Hepatocellular 

Carcinoma: Impact of Liver Dysfunction and Development 

of a Predictive Statistical Model: an International 

Collaborative Project 

Imam Waked 1 , Sarah Berhane 2 , Stephen L. Chan 3 , Asmaa 

Gomaa 4 , Mercedes Iñarrairaegui 5 , Zahraa Alkhatib 4 , Takashi 

Kumada 6 , Paul B. Lai 7 , Tim Meyer 8 , Frankie Mo 3 , Daniel Palmer 2 , 

Bruno Sangro 5 , Nicholas Stern 9 , Toshifumi Tada 6 , Hidenori 

Toyoda 6 , Arndt Vogel 10 , Winnie Yeo 3 , Phillip Johnson 2 ; 1 Hepatology, 

National Liver Institute, Menoufiya, Egypt; 2 Molecular 

and Clinical Cancer Medicine;, University of Liverpool, Liverpool, 

United Kingdom; 3 Sir Y. K. Pao Centre for Cancer, Chinese University 

of Hong Kong, Hong Kong, China; 4 Oncology, National 

Liver Institute, Shebeen El Kom, Egypt; 5 Centro de Investigacion 

Biomedica en Red de Enfermedades Hepaticas y Digestivas, Clinica 

Universidad de Navarra, Pamplona, Spain; 6 Ogaki Municipal 

Hospital, Ogaki, Japan; 7 Surgery, The Chinese University of Hong 

Kong, Hong Komg, China; 8 Oncology, University College of London 

Cancer Institute, London, United Kingdom; 9 Gastroenterology, 

Aintree University Hospital, Liverpool, United Kingdom; 10 Hannover 

Medical School, Hannover, Germany 

Background: TransArterial Chemo-Embolisation (TACE) is recommended 

for patients with BCLC intermediate stage HCC 

(stage B) particularly in patients with excellent underlying liver 

function and minimal symptoms. A recently developed measure 

of liver function (the Albumin-Bilirubin (ALBI) grade) now permits 

detailed assessment of the impact of liver function on survival 

(1). Methods: We accrued patient level data from 2454 

patients undergoing TA(C)E – China (n= 242), Japan (n=655), 

Europe (UK, Germany and Spain combined, n=559 ) and 

Egypt (n=998). Overall, 59 % had Child-Pugh grade ‘A’ and 

17%, portal vein invasion. Liver function was graded according 

to their ALBI grade. A statistical model for survival after 

TA(C)E was built on a randomly selected half of a combined 

group of 1214 Japanese and European patients, n=602 (323 

Japanese and 279 European) and then validated the on the 

remaining 612 patients (332 Japanese and 280 European). 

The TACE model was also validated on an independent cohort 

from Egypt (n=998). Results: Our TACE model accurately predicted 

survival in the second half validation set (actual median 

survival = 20.4 months, predicted = 20.7 months) as well as 

in the Egyptian cohort (actual median survival = 18 months, 

predicted = 20 months). One year survival was 65.7 and 

71.3% for actual and predicted respectively). Classification of 

patients according their ALBI grade resulted in clear, non-overlapping 

survival curves in all cohorts (figure) Conclusion: ALBI 

categorised patients receiving TACE into three clear prognostic 

groups. Our TACE model accurately predicted survival on the 

basis of baseline clinical and laboratory features. References 

1. Johnson PJ et al., J Clin Oncol. JCO.2014.57.9151


Multivariate Cox proportional hazards model analysis for risk of 

overall mortality 

Disclosures: 

The following authors have nothing to disclose: Teng-Yu Lee, Yi-Ju Chen, Jaw- 

Town Lin, Hsiu-Jon Ho, Ming-Shiang Wu, Chun-Ying Wu 

Disclosures: 

Imam Waked - Advisory Committees or Review Panels: Janssen; Speaking and 

Teaching: Hoffman L Roche, Merck, BMS, Gilead, AbbVie 

Bruno Sangro - Speaking and Teaching: Sirtex Medical Europe, Bayer Schering 

Pharma 

Nicholas Stern - Speaking and Teaching: Bayer Pharmaceuticals 

The following authors have nothing to disclose: Sarah Berhane, Stephen L. Chan, 

Asmaa Gomaa, Mercedes Iñarrairaegui, Zahraa Alkhatib, Takashi Kumada, 

Paul B. Lai, Tim Meyer, Frankie Mo, Daniel Palmer, Toshifumi Tada, Hidenori 

Toyoda, Arndt Vogel, Winnie Yeo, Phillip Johnson 

345 

Nucleos(t)ide analogue therapy in relation to survival 

after transarterial chemoembolization for HBV-related 

hepatocellular carcinoma 

Teng-Yu Lee 1 , Yi-Ju Chen 1 , Jaw-Town Lin 2 , Hsiu-Jon Ho 2 , Ming-Shiang 

Wu 3 , Chun-Ying Wu 1 ; 1 Taichung Veterans General Hospital, 

Taichung, Taiwan; 2 Fu Jen Catholic University, New Taipei City, 

Taiwan; 3 National Taiwan University Hospital, Taipei, Taiwan 

Transarterial chemoembolization (TACE) is the most widely 

used primary treatment for unresectable hepatocellular carcinoma 

(HCC), but patient survival is unsatisfactory. We aimed 

to investigate the association between nucleos(t)ide analogue 

(NA) therapy and patient survival in hepatitis B virus (HBV)-related 

HCC following TACE. Using the Taiwan National Health 

Insurance Research Database between October 1, 2003 and 

December 31, 2011, we screened 57,566 patients with newly 

diagnosed HCC. Excluding patients with advanced HCC, we 

identified 5,182 patients who received TACE for HBV-related 

HCC (869 used NAs 90 days and 4,313 never used NA 

after TACE). Patients in the NA-treated cohort were randomly 

matched 1:4 with patients in the untreated cohort by age, sex, 

cirrhosis, coexisting diseases and the time period between 

TACE and initiation of NA therapy. Finally, 582 patients were 

recruited in the NA-treated group and 2,328 in the untreated 

group. Cumulative incidences of and hazard ratios (HRs) for 

patient mortality were analyzed. The mortality rates of the 

NA-treated group were significantly lower than those of the 

untreated group (1-year: 43.5%, 95% confidence interval [CI]: 

39.3-47.7% vs. 56.8%, 95%CI: 54.7-58.9%; 2-year: 58.4%, 

95%CI: 53.9-62.9% vs. 74.5%, 95%CI: 52.3-76.6%; 5-year: 

78.4%, 95%CI: 73.2-83.6% vs. 88.6%, 95%CI: 86.3-90.8%; 

P< 0.001). In multivariate regression analysis, NA therapy was 

independently associated with a decreased mortality risk (HR 

0.66, 95%CI: 0.59-0.75; P< 0.001). Multivariate stratified 

analyses verified the association of NA therapy and decreased 

mortality in all patient subgroups. Conclusion: NA therapy was 

associated with a decreased mortality among patients with 

HBV-related HCC following TACE. 

346 

Risk assessment of hepatocellular carcinoma in chronic 

hepatitis B patients with long-term antiviral therapy 

Kyu sik Chung, Do Young Kim, Beom Kyung Kim, Seung Up Kim, 

Jun Yong Park, Hye Jin Ku, Kwang-Hyub Han, Sang Hoon Ahn; 

Department of Internal Medicine, Yonsei University College of 

Medicine, Seoul, Korea (the Republic of) 

Background: In the era of potent antiviral therapy, the prognostic 

significance of a biological gradient of serum HBV-DNA levels 

measured at given time points has substantially diminished, 

another predictive factors, determining the long-term prognosis 

such as hepatocellular carcinoma (HCC) development, are 

required Methods: The aim of this study was to evaluate the 

incidence and risk factor of HCC occurrence in a large group 

of treatment-naïve chronic hepatitis B (CHB) patients, treated 

with entecavir (ETV) or tenofovir (TDF). Additionally, predictive 

accuracy of published HBV-HCC risk scores (CU-HCC 

score, GAG-HCC score, REACH-B score and LSM-HCC score) 

were evaluated. Results: A total of 762 CHB patients, who 

were treated with ETV (n=546) or TDF (n=216), were consecutively 

enrolled in this study. The median age (486 males and 

276females) was 51 years and 261 (34.3%) patients have 

liver cirrhosis. During the median follow-up of 45.8 months, 

HCC developed in 39 patients (5.1%) and the 1-, 3-, 5-, and 

7-year cumulative incidences of HCC were 1.6%, 4.0%, 7.2% 

and 9.5%, respectively. In multivariate analysis, old age (hazard 

ratio [HR] 1.091; 95% confidence interval [CI], 1.040- 

1.144;P0.05). In terms of HCC 

development at 3-/5-years, the AUROC of CU-HCC score were 

0.851/0.846, LSM-HCC score, 0.772/0.763, GAG-HCC 

score, 0.719/0.739, and REACH-B score, 0.710/0.691, indicating 

that liver fibrotic burden need to be incorporated properly 

into HBV-HCC prediction models to improve predictive 

performance. Conclusions: In the era of antiviral therapy, individual 

risks of HBV-HCC should be assessed effectively based 

on age and the fibrotic burden, not serum HBV-DNA level or 

virological response to antiviral therapy. 

Disclosures: 

The following authors have nothing to disclose: Kyu sik Chung, Do Young Kim, 

Beom Kyung Kim, Seung Up Kim, Jun Yong Park, Hye Jin Ku, Kwang-Hyub Han, 

Sang Hoon Ahn


347 

Mutations of p53, ARID1A and KRAS mutations and 

their association with “subtypes with stem-cell feature” 

in combined hepatocellular-cholangiocellular carcinoma 

Motoko Sasaki 1 , Yasunori Sato 1 , Yasuni Nakanuma 2,1 ; 1 Human 

Pathology, Kanazawa University Graduate School of Medicine, 

Kanazawa, Japan; 2 Shizuoka Cancer Center, Shizuoka, Japan 

Backgrounds/Aims. Combined hepatocellular-cholangiocarcinoma 

(cHC-CC) is composed of hepatocellular carcinoma 

(HCC), cholangiocarcinoma (CC) and diverse components with 

intermediate features between HCC and CC and is characterized 

by poor prognosis. Subtypes with stem cell features (SC 

subtype) including typical subtype (TS), intermediate cell subtype 

(INT) and cholangiolocellular type (CLC) were proposed 

in the WHO classification 2010. We have previously reported 

that each SC subtype may have different clinicopathological 

significance in cHC-CC. That is, the proportion of INT was significantly 

correlated to histological grading of coexistent HCC 

and tumor size, whereas the proportion of CLC was inversely 

correlated to histological grading of coexistent HCC and 

tumor size. In this study, we examined mutation status of p53, 

ARID1A and KRAS and their association with clinicopathological 

features, especially with SC subtypes, in cHC-CC. Methods. 

Fifty-three patients with cHC-CC (14 women and 39 men, age 

ranged 36-83 yrs, mean±SD, 65± 9.5yrs) were retrieved from 

our pathological files (1996-2014). The background diseases 

were hepatitis B (n=9), hepatitis C (21), chronic alcoholism or 

nonalcoholic fatty liver disease (NAFLD) (8) and cryptogenic 

(15). Mutations of p53, ARID1A and KRAS were also evaluated 

using immunohistochemistry (p53, ARID1A) and direct 

sequencing (KRAS). The prevalence of each component (HCC, 

TS, INT, CLC and CC) was histologically assessed with assistance 

of mucin staining and immunohistochemical staining 

for CK7, CK19, EMA, EpCAM, NCAM, AFP and HepPar1. 

Results: Mutations of p53, ARID1A and KRAS were detected 

in 24 cases (45%), 7 (13%) and 4 of 39 (10%) respectively. 

Mutations of both p53 and ARID1A were detected in 3 and 

mutations of both p53 and KRAS were detected in 2 cHC- 

CCs. SC subtypes were observed in all cHC-CCs in various 

amount and combination. The prevalence of each SC subtype 

in cHC-CC was as follows; TS, 8 (15%); INT, 33 (62%); and 

CLC, 35 (66%). ARID1A-mutated cHC-CCs were significantly 

smaller size (p


The following authors have nothing to disclose: Suraj Sharma, Matthew Kowgier, 

Bettina E. Hansen, Korosh Khalili, Willem Pieter Brouwer, Gideon Hirschfield 

The following authors have nothing to disclose: Yee-Kit Tse, Terry C. Yip 

349 

Oral nucleos(t)ide analogues (NAs) improve survival 

of patients with hepatocellular carcinoma after tumor 

resection but not after other treatment modalities – a 

study of 2,198 patients with chronic hepatitis B 

Grace LH Wong, Yee-Kit Tse, Vincent W. Wong, Terry C. Yip, 

Henry Lik-Yuen Chan; Institute of Digestive Disease, The Chinese 

University of Hong Kong, Shatin, Hong Kong 

Background: Oral nucleos(t)ide analogues (NAs) effectively 

suppress hepatitis B virus (HBV) and reduce tumor recurrence 

and death in patients with HBV-related hepatocellular carcinoma 

(HCC). We aimed to investigate the effect of NAs on 

the clinical outcomes after different HCC treatments (tumor 

resection, local ablative therapy [LAT], transartierial chemoembolization 

[TACE]) for HCC in these patients. Methods: We 

conducted a territory-wide cohort study using the database 

from Hospital Authority, which provides medical services at 

both in-patient and out-patient settings for 70-80% of the Hong 

Kong citizens. We identified chronic hepatitis B (CHB) patients 

with HCC by International Classification of Diseases, Ninth 

Revision, Clinical Modification (ICD-9-CM) diagnosis codes, 

diagnosed between 2000 and 2012. HCC treatments, NA 

use and laboratory parameters were retrieved and studied. 

The primary outcome was HCC recurrence and death. A 

3-month landmark analysis was used to evaluate the relative 

risk of primary outcome in patients with or without NA treatment. 

Cox proportional hazards models were used to estimate 

adjusted HRs with 95% CIs of HCC recurrence (taking death 

into account as a competing risk) and mortality associated 

with post-treatment use of NA. Results: 2,198 CHB patients 

(1,230 NA-untreated and 968 NA-treated) with HCC received 

at least one type of HCC treatment were included in the analysis. 

Tumor resection, LAT, TACE, resection-LAT and resection/ 

LAT-TACE were the HCC treatments in 810 (36.9%), 1,015 

(46.2%), 222 (10.1%), 73 (3.3%) and 78 (3.5%) patients. 

At a median follow-up of 2.8 years (IQR 1.4 – 4.9 years)”, 

tumor recurrence and death occurred in 451 (36.7%) and 578 

(47.0%) of NA-untreated subjects; and 216 (22.3%) and 301 

(31.1%) of NA-treated subjects. NA therapy reduced the risk of 

overall HCC recurrence (adjusted sub-hazard ratio [SHR] 0.62, 

95% confidence interval [CI] 0.49–0.80; P 60 

years old). Results: Surveillance starts to be effective above 45 

years of age (gaining >100 quality-adjusted life days) though 

it is still relatively cost-ineffective with an incremental cost-effectiveness 

ratio (ICER) of $88,160/QALY. As the starting age 

of surveillance increases, the ICER for surveillance decreases. 

Beyond the age of 58, the ICER for surveillance is consistently 

lower than the standard willingness-to-pay threshold of 

$50,000/QALY. Conclusions: Due to the marked decrease in 

HCC incidence, universal screening in patients with cirrhosis 

post-SVR is very unlikely to be cost-effective; however, by targeting 

surveillance to those over 55 years of age (figure), HCC 

surveillance would represent better value-for-money. Data to 

identify high-risk populations may allow for further tailoring of 

recommendations according to a patient’s risk profile.


Disclosures: 

Adriaan J. van der Meer - Consulting: Gilead; Speaking and Teaching: MSD, 

Gilead 






Jordan J. Feld - Advisory Committees or Review Panels: Merck, Janssen, Gilead, 

AbbVie, Theravance, Bristol Meiers Squibb; Grant/Research Support: AbbVie, 

Boehringer Ingelheim, Janssen, Gilead, Merck 

The following authors have nothing to disclose: Hooman Farhang Zangneh, William 

W. Wong, Beate Sander, Chaim M. Bell, Khalid Mumtaz, Matt Kowgier, 

Sean P. Cleary, Kelvin K. Chan 

351 

Whole blood coagulation tests are not equally able to 

detect haemostatic prothrombotic alterations in patients 

with liver cirrhosis and hepatocellular carcinoma (HCC): 

rotational thromboelastometry versus thrombingeneration 

test. 

Alberto Zanetto 1 , Alberto Ferrarese 1 , Alessandro Vitale 2 , Umberto 

Cillo 2 , Mariangela Fadin 3 , Sabrina Gavasso 3 , Claudia M. Radu 3 , 

Luca Spiezia 3 , Fabio Farinati 1 , Patrizia Burra 1 , Paolo Simioni 3 , 

Marco Senzolo 1 ; 1 Surgery, Oncology and Gastroenterology, 

University of Padua, Gastroenterology, Padova, Italy; 2 Surgery, 

Oncology and Gastroenterology, University of Padua, Liver Transplantation 

Unit, Padova, Italy; 3 Department of Medicine, Padua 

University Hospital, V Chair of Internal Medicine, Padova, Italy 

Background and aim: neoplasms are considered as one of the 

main important cause of acquired thrombophilia but studies 

exploring coagulation imbalance induced by HCC in liver cirrhosis 

are lacking. The aim of this study study was to evaluate the 

thrombophilic role of HCC in patients with cirrhosis. Methods: 

cirrhotic patients with and without HCC were enrolled in the 

study and underwent rotational thromboelastometry (ROTEM), 

platelet count, determination of levels of pro and anticoagulation 

factors and thrombin generation test (TG), with and without 

trombomodulin (TM). Results: 76 cirrhotics, of whom 41 with 

HCC, were included. Volume of active HCC was >5 cm 3 in 

18 patients. Levels of pro and anticoagulation factors were 

similar in patients with and without HCC, but fibrinogen was 

increased in HCC patients with tumor volume >5cm 3 compared 

to those with 5cm 3 compared to those with


that mediates doxorubicin sensitivity through a process that 

involves its acetylation and S574 phosphorylation. SIRT6 can 

deacetylate FOXO3 preventing apoptosis, and TACE-resistant 

tumors have higher SIRT6 levels. Manipulation of FOXO3 modifications 

may thus prove useful in enhancing the chemotherapy 

sensitivity of HCC. 

Disclosures: 

Steven A. Weinman - Consulting: Cardax, Inc. 

The following authors have nothing to disclose: Josiah Cox, Zhuan Li, Anusha 

Vittal, Maura O’Neil, Brian Bridges, Sean Kumer, Timothy Schmitt 

353 

Long non-coding RNA integrator complex subunit 6 

pseudogene 1 (INTS6P1), a tumor suppressor and prognostic 

factor in hepatocellular carcinoma, induces apoptosis 

via intrinsic mitochondrial pathway 

Minqiang Lu 1 , Ka Yin LUI 1 , Haoran Peng 1,2 , Rongdang Fu 1 , Huan 

Chen 1 , Chunhui Qiu 1 ; 1 Hepatic Surgery, The Third Affiliated Hospital 

of Sun Yat-sen University, Guangzhou, China; 2 Division of 

Gastroenterology and Hepatology, The Johns Hopkins Hospital, 

Baltimore, MD 

Purpose: Long non-coding RNAs (lncRNAs) have been proved 

its close association with many diseases especially with tumor. 

We aimed to determine whether the lncRNA integrator complex 

subunit 6 pseudogene 1 (INTS6P1) could be as a tumor 

suppressor and its mechanism in hepatocellular carcinoma 

(HCC). Materials and methods: Firstly, the expression level of 

INTS6P1 mRNA were measured in a cohort of 60 HCC tissues 

and adjacent normal liver tissues by using the quantitative 

real time-polymerase chain reaction (qRT-PCR); Secondly, the 

functional studies of INTS6P1, include growth curves, migration 

assays and cell death, were detected in HCC cell lines. 

Finally, the mechanism experiment was investigated for tumor 

suppressive roles of INTS6P1. Results: We found that lncRNA 

INTS6P1 was remarkably down-regulated in HCC (71.4%), 

and its expression was significantly correlated with pathology 

grade (p


355 

Prognostic assessment of patients with intermediate 

stage, untreated hepatocellular carcinoma 

Edoardo G. Giannini 1 , Alessandro Moscatelli 1 , Gaia Pellegatta 1 , 

Fabio Farinati 2 , Francesca Ciccarese 3 , Fabio Piscaglia 4 , Gian 

Ludovico Rapaccini 5 , Mariella Di Marco 6 , Eugenio Caturelli 7 , 

Marco Zoli 8 , Franco Borzio 9 , Giuseppe Cabibbo 10 , Martina M. 

Felder 11 , Rodolfo Sacco 12 , Filomena Morisco 13 , Gabriele Missale 

14 , Francesco G. Foschi 15 , Antonio Gasbarrini 16 , Gianluca 

Svegliati Baroni 17 , Roberto Virdone 18 , Maria Chiaramonte 19 , 

Franco Trevisani 20 ; 1 Dipartimento di Medicina Interna, Unità di 

Gastroenterologia, IRCCS-Azienda Ospedaliera Universitaria San 

Martino-IST, Università di Genova, Genova, Italy; 2 Dipartimento di 

Scienze Chirurgiche e Gastroenterologiche, Unità di Gastroenterologia, 

Università di Padova, Padova, Italy; 3 Divisione di Chirurgia, 

Policlinico San Marco, Zingonia, Italy; 4 Dipartimento di Scienze 

Mediche e Chirurgiche, Unità di Medicina, Alma Mater Studiorum 

– Università di Bologna, Bologna, Italy; 5 Unità di Medicina Interna 

e Gastroenterologia, Complesso Integrato Columbus, Università 

Cattolica di Roma, Roma, Italy; 6 Divisione di Medicina, Azienda 

Ospedaliera Bolognini, Seriate, Italy; 7 Unità Operativa di Gastroenterologia, 

Ospedale Belcolle, Viterbo, Italy; 8 Dipartimento 

di Scienze Mediche e Chirurgiche, Unità di Medicina Interna, 

Alma Mater Studiorum – Università di Bologna, Bologna, Italy; 

9 Dipartimento di Medicina, Unità di Radiologia, Ospedale Fatebenefratelli, 

Milano, Italy; 10 Dipartimento Biomedico di Medicina 

Interna e Specialistica, Unità di Medicina Gastroenterologia, 

Università di Palermo, Palermo, Italy; 11 Ospedale Regionale 

di Bolzano, Unità di Gastroenterologia, Bolzano, Italy; 12 Unità 

Operativa Gastroenterologia e Malattie del Ricambio, Azienda 

Ospedaliero-Universitaria Pisana, Pisa, Italy; 13 Dipartimento di 

Medicina Clinica e Chirurgia, Unità di Gastroenterologia, Università 

di Napoli “Federico II”, Napoli, Italy; 14 Unità di Malattie Infettive 

ed Epatologia, Azienda Ospedaliero-Universitaria di Parma, 

Parma, Italy; 15 Dipartimento di Medicina Interna, Ospedale per 

gli Infermi di Faenza, Faenza, Italy; 16 Unità di Medicina Interna 

e Gastroenterologia, Policlinico Gemelli, Università Cattolica di 

Roma, Roma, Italy; 17 Clinica di Gastroenterologia, Università 

Politecnica delle Marche, Ancona, Italy; 18 Dipartimento Biomedico 

di Medicina Interna e Specialistica, Unità di Medicina Interna 

2, Azienda Ospedaliera Ospedali Riuniti Villa Sofia-Cervello, Palermo, 

Italy; 19 Unità di Gastroenterologia, Ospedale Sacro Cuore 

Don Calabria, Negrar, Italy; 20 Dipartimento di Scienze Mediche 

Chirurgiche, Unità di Semeiotica Medica, Alma Mater Studiorum 

– Università di Bologna, Bologna, Italy 

Background: The Barcelona Clinic Liver Cancer (BCLC) intermediate 

stage (BCLC B) includes a heterogeneous population 

of patients with hepatocellular carcinoma (HCC), who often 

display different survival rates. Due to these reasons, a panel 

of experts recently proposed a sub-classification of the intermediate 

stage HCC in order to facilitate prognostic assessment 

and treatment decisions in clinical practice. Aim: In this study 

our aim was to assess the prognostic capability of the re-classification 

of the intermediate stage HCC (BCLC B) in a large 

cohort of patients with untreated HCC managed by the Italian 

Liver Cancer (ITA.LI.CA) Group. Methods: We assessed the 

prognosis of 269 untreated patients with HCC observed in the 

period 1987-2012, and who were classified according to the 

proposed sub-classification of the BCLC B stage from stage B1 

to stage B4 (Semin Liver Dis 2012;32:348-59). Survival was 

defined as the time, expressed in months, that elapsed from 

the date of HCC diagnosis and the date of death, or of the 

most recent follow-up information. Results: According to the 

proposed sub-classification of the intermediate stage HCC, 65 

patients were classified B1 (24.2%), 105 patients B2 (39.0%), 

22 patients B3 (8.2%), and 77 patients B4 (28.6%). Overall 

median survival was 13 months, and progressively decreased 

from stage B1 (25 months) through stages B2 (16 months) 

and B3 (9 months), to stage B4 (5 months, P


B3, C), alpha-fetoprotein, performance status and Child–Pugh 

score in predicting survival and guiding treatment allocation. 

An ITA.LI.CA prognostic score and treatment scheme were then 

constructed by integration of clinical judgments, and then compared 

with other systems (BCLC, HKLC, MESIAH, CLIP, JIS). 

Findings. Median survival was well stratified by the main ITA. 

LI.CA stages (p


(r=0.68). exRNA aligned read counts were lower in cancer 

than in normal bile samples. Read counts between cancer and 

normal bile exRNA samples had a wide range of correlation (r: 

0.64 to 0.95). A set of 14 mature (sense and antisense) miRNA 

were differentially expressed > 3SD fold change in expression 

(based on reads per million) in bile from CCA compared with 

normal bile. Other differentially expressed exRNA identified 

included tRNA and piRNA. Summary and Conclusions: Biliary 

exRNA sequencing provides a novel strategy for biomarker 

discovery. exRNA isolation protocols have a major impact on 

yield and quality of exRNA for downstream gene expression 

studies. Differentially expressed candidate exRNA biomarkers 

have been identified that may be useful for the diagnosis of 

CCA. 

Disclosures: 

The following authors have nothing to disclose: Irene K. Yan, Waseem David, 

Swathi Mohankumar, Sarah Nix, Hiroaki Haga, Yan W. Asmann, Tushar Patel 

359 

Risk Differences of Hepatocellular Carcinoma (HCC) 

Among Patients with Chronic Hepatitis C (CHC) of Different 

Viral Genotypes: a Historical Cohort Study in the 

United States 

Mingjuan Jin 2,1 , Changqing Zhao 1,7 , Joseph K. Hoang 1 , Nghia 

H. Nguyen 1,3 , An K. Le 1 , Christine Y. Chang 1 , Richard H. Le 1 , 

Alina Kutsenko 4 , Lee Ann Yasukawa 5 , Jian Q. Zhang 6 , Susan C. 

Weber 5 , Mindie H. Nguyen 1 ; 1 Division of Gastroenterology and 

Hepatology, Stanford University Medical Center, Palo Alto, CA; 

2 Department of Epidemiology and Biostatistics, School of Public 

Health, Zhejiang University, Hangzhou, China; 3 Department of 

Medicine, University of California, San Diego, San Diego, CA; 

4 Department of Medicine, Stanford University Medical Center, 

Palo Alto, CA; 5 Center for Clinical Informatics, Stanford University 

School of Medicine, Palo Alto, CA; 6 Chinese Hospital, San Francisco, 

CA; 7 Department of Cirrhosis, Institute of Liver Disease, Shuguang 

Hospital, Shanghai University of T.C.M., Shanghai, China 

Purpose: Whether there exists differences in the risk for HCC 

among patients with CHC of different genotypes is still controversial, 

which were explored in the present study in a historical 

cohort from 2 ethnically diverse US centers. Methods: 

The subject inclusion was as follows: CHC patients aged 20 

years or older, presented at study centers between 1/1/1999 

and 12/31/2013, followed up for at least 1 year, having 

HCV genotype information, and no HCC at baseline. The HCC 

cumulative incidence and incidence density were calculated. 

Cox proportional hazards models were used to testify the associations 

of HCV genotypes with HCC. Results: A total of 1990 

eligible CHC patients were included, among whom 1174 

(59.0%) were female and 1594 (80.1%) were ≤ 60 years old 

with mean age of 53.20±9.63 years. The ethnicity distribution 

was 7.9% for Asian, 40.6% for White, 18.3% for Hispanc and 

Black, and 33.2% for others and unknown. The frequencies of 

genotypes 1a/untyped, 1b, 2, 3, and others (genotype 4 and 

6) were 51.4%, 18.6%, 13.0%, 13.1%, and 3.9%, respectively. 

Overall, the 1-year, 3-year and 5-year HCC cumulative 

incidence were 448.53, 1341.88, and 2354.31 per 10,000 

persons, respectively, with the incidence density of 297.37 

per 10, 000 person-years in this cohort. Compared to patients 

with the most prevalent genotype 1a/untyped, the age-, gender- 

and ethnicity-adjusted hazard ratios (95%CIs) were 0.88 

(0.64-1.22), 0.58 (0.37-0.93), 1.42 (0.99-2.04), and 1.89 

(1.14-3.14) for those with genotype 1b, 2, 3, and 4 and 6, 

respectively. Conclusions: The results suggest that HCV genotype 

4 and 6 are associated with the highest risk of HCC, and 

genotype 2 has the lowest risk, and there were no significant 

differences between genotype 1b and 1a/untyped . Preventive 

measures such as antiviral therapy and HCC surveillance 

efforts should especially target the higher risk groups. 

Disclosures: 





The following authors have nothing to disclose: Mingjuan Jin, Changqing Zhao, 

Joseph K. Hoang, Nghia H. Nguyen, An K. Le, Christine Y. Chang, Richard H. 

Le, Alina Kutsenko, Lee Ann Yasukawa, Jian Q. Zhang, Susan C. Weber 

360 

Surveillance for Hepatocellular Carcinoma (HCC) Using 

Ultrasonography Alone is Associated with Improved 

Survival: Results of Inverse Probability of Treatment 

Weighting (IPTW)-Based Survival Analysis 

Piyanat Chattieng 2 , Jonggi Choi 1 , Nopavut Geratikornsupuk 2 , 

Kessarin Thanapirom 2 , Sombat Treeprasertsuk 2 , Piyawat Komolmit 

2 , Roongruedee Chaiteerakij 2,1 ; 1 Mayo Clinic, Rochester, MN; 

2 Medicine, Chulalongkorn University, Bangkok, Thailand 

Background/Aim: The current American Association for the 

Study of Liver Disease guideline recommend HCC surveillance 

program using ultrasound modality alone. The impact of the surveillance 

program on survival remains unclear due to unavoidable 

bias and confounders in observational studies. We aimed 

to determine the effect of HCC surveillance on survival using 

an IPTW analysis controlling for inherent bias and confounders 

existing in observational studies. Methods: We conducted a 

retrospective cohort study of 446 patients with cirrhosis and/ 

or chronic hepatitis B infection who were diagnosed with HCC 

between 2007 and 2013 in a major referral center in Bangkok, 

Thailand. Surveillance was defined as having repeated 

abdominal ultrasonography within 1 year prior to HCC diagnosis. 

Survivals of patients with and without surveillance were 

estimated using the Kaplan-Meier method with lead-time bias 

adjustment and compared using the log-rank test. Hazard ratio 

(HR) and 95% confidence interval (CI) of surveillance was 

computed using conventional Cox and weighted Cox proportional 

hazards analysis with IPTW adjustment. Results: Of the 

446 HCC patients, 79% were male with a mean age of 58.1 

years. Hepatitis B (54%), hepatitis C (20%), alcohol (11%) and 

non-alcoholic fatty liver disease (9%) were the most common 

underlying chronic liver diseases. A hundred and three (23%) 

were diagnosed with HCC through surveillance. The surveillance 

group had a higher proportion of patients diagnosed 

with the Barcelona-Clinic Liver Cancer (BCLC) stage A (82% vs. 

34%, P


P=0.005. The estimated effect of surveillance remained similar 

using IPTW-adjusted Cox analysis, with HR (95% CI) of 

0.57 (0.43-0.76), P


363 

Prediction of Recurrence after Radiofrequency Ablation 

of Hepatocellular Carcinoma using Liver Stiffness Measurement 

(FibroScan) 

Yu Rim Lee 1 , Soo Young Park 1 , Se Young Jang 1 , Su Hyun Lee 1 , 

Sun Kyung Jang 1 , Won Young Tak 1 , Young Oh Kweon 1 , Jung Gil 

Park 2 , Seung Up Kim 3 ; 1 Kyunopook National University Hospital, 

Daegu, Korea (the Republic of); 2 Internal Medicine, Gastroenterology 

and Hepatology, Gumi, Korea (the Republic of); 3 Internal 

Medicine, Gastroenterology, Seoul, Korea (the Republic of) 

Background. Hepatocellular carcinoma(HCC) is a third leading 

cause of cancer related death and incidence is increasing 

every year. As active surveillance programs develop, detection 

of early stage HCC are increasing. RFA as well as hepatic 

resection and liver transplantation have been considered 

standard treatments of HCC. However, long-term prognosis 

is unsatisfactory due to HCC recurrence associated with liver 

fibrosis. The purpose of this study is to predict recurrence after 

radiofrequency ablation of hepatocellular carcinoma using 

liver stiffness measurement(LSM), which is known related to be 

effective in evaluating the liver fibrosis noninvasively. Methods. 

A total of 131 patients who performed LSM before RFA 

between November, 2008 and December, 2014(training set) 

and 55 between November, 2006 and December, 2012(validation 

set) was enrolled. Multiple clinical variables including 

LSM were analysed for association with recurrence. The result 

was applied to validation set. Results. Baseline characteristics 

including alfa-fetoprotein, INR, Child-Pugh Class(A/B), underlying 

liver disease(B/C/nonB,nonC/Alcohol) and recurrence 

rate of training set and validation set were different(p 12.3 kPa had shorter disease free survival 

in training set. In validation set, using a cutoff value of LSM 

12.3 kPa, recurrence rate were also significantly greater at 

LSM values > 12.3 kPa patients(p


365 

When to perform surgical resection or radiofrequency 

ablation for early hepatocellular carcinoma? A nomogram-guided 

treatment strategy 

Po-Hong Liu 1,2 , Chia-Yang Hsu 2,3 , Yi-Hsiang Huang 1,4 , Chien- 

Wei Su 1,2 , Han-Chieh Lin 1,2 , Teh-Ia Huo 1,5 ; 1 Department of Medicine, 

Taipei Veterans General Hospital, Taipei, Taiwan; 2 Faculty 

of Medicine, National Yang-Ming University, Taipei, Taiwan; 

3 Department of Internal Medicine, University of Nevada School 

of Medicine, Reno, NV; 4 Institute of Clinical Medicine, National 

Yang-Ming University, Taipei, Taiwan; 5 institute of Pharmacology, 

National Yang-Ming University, Taipei, Taiwan 

Background&Aims: Radiofrequency ablation (RFA) is indicated 

for early hepatocellular carcinoma (HCC). The comparative 

efficacy between RFA and surgical resection (SR) is inconclusive. 

We aimd to develop a prognostic nomogram for recurrence-free 

survival (RFS) after RFA and to evaluate its ability 

in improving treatment algorithm. Methods: We enrolled 836 

patients with BCLC very early/early HCC receiving SR or RFA. 

Nomogram was constructed with Cox proportional hazards 

model. RFA patients were stratified into low-risk and high-risk 

groups. The RFS and overall survival (OS) of two risk groups 

were compared with SR patients by propensity score matching 

analysis. Results: A prognostic nomogram was built based 

on number and size of tumor, platelet count, albumin level, 

and model for end-stage liver disease score with a c-index of 

0.69. SR provided better RFS and OS compared with high-risk 

(nomogram score ≥9.8) RFA patients in the propensity model. 

The 5-year RFS rates were 36% vs. 11%, while 5-year OS rates 

were 74% vs. 60% for SR and high-risk RFA group, respectively 

(both p


367 

Meta-Analysis: Proportions of Hepatocellular Carcinoma 

(HCC) Diagnosed by Screening, Surveillance, or Without 

Symptoms is Dismally Low Across Different World 

Regions and Underlying Liver Diseases 

Michelle Q. Jin 1 , Richard H. Le 1 , Mingjuan Jin 1,2 , Changqing 

Zhao 1,3 , Michael H. Le 4 , Vincent L. Chen 1 , Grace Wong 5 , Vincent 

W. Wong 5 , Young-Suk Lim 6 , Wan-Long Chuang 7 , Ming-Lung Yu 7 , 

Mindie H. Nguyen 1 ; 1 Division of Gastroenterology and Hepatology, 

Stanford University Medical Center, Palo Alto, CA; 2 Department 

of Epidemiology and Biostatistics, School of Public Health, 

Zhejiang University, Hangzhou, China; 3 Department of Cirrhosis, 

Institute of Liver Disease, Shuguang Hospital, Shanghai University 

of T.C.M, Shanghai, China; 4 University of California, Santa Cruz, 

Santa Cruz, CA; 5 Division of Gastroenterology and Hepatology, 

Department of Medicine and Therapeutics, The Chinese University 

of Hong Kong, Hong Kong, China; 6 Department of Gastroenterology, 

Asan Liver Center, Asan Medical Center, University of Ulsan 

College of Medicine, Seoul, Korea (the Republic of); 7 Department 

of Internal Medicine, Kaohsiung Medical University Hospital, 

Kaohsiung Medical University, Kaohsiung, Taiwan 

Background: Guidelines for HCC screening and surveillance 

are available worldwide, but poor adherence to guidelines 

may result in symptomatic diagnosis with late-stage disease. 

Our goal is to estimate the proportions of HCC patients diagnosed 

via screening, surveillance or at asymptomatic stage 

(asymptomatic/screening HCC). Methods: PubMed search 

with MEDLINE filter for “HCC,” “Screening/Surveillance” and 

associated terms was performed in March 2015. Abstracts 

from recent international liver meetings were also reviewed. 

Two investigators identified original studies published in or 

after 2000 that reported proportions of asymptomatic/screening 

HCC with ≥100 HCC patients (except for subanalysis). 

We performed analysis using a random-effects model. Results: 

A total of 1356 studies were initially identified and full paper 

review was performed for 128 studies, yielding 28 eligible 

studies with 15,244 HCC patients. The overall pooled proportion 

of asymptomatic/screening HCC patients was 34% 

(95% CI 27-40%), and it was significantly lower in Asia-Pacific 

(23%, 95% CI 16-31%) compared to Europe (39%, 95% CI 

28-50%) (p


compared to those with score


needle-aspiration biopsy specimens of 44 HCC patients (20 

in the HBV group, 19 in the HCV group and 5 in the NASH 

group) were subjected to real-time PCR to measure expression 

levels of TAA mRNAs in tumor tissues. Results IFN-g ELISpot 

assay revealed that immune responses to the p53-derived epitope 

(positive rates: 20.7% in the HBV group, 5.0% in the HCV 

group and 0% in the NASH group), and to the MRP3-derived 

epitope (positive rates: 20.7% in the HBV group, 2.5% in the 

HCV group and 5.6% in the NASH group) were significantly 

induced in the HBV group than in other groups (p


median survival (MS) significantly improved from 1990-2011 

in localized, regional and distant metastases with CDS. In the 

65-75 age population MS improved only in CDS localized 

disease; 7 months (1990-2000) to 9.8 months (2001-2011). In 

the 75+ age population, the MS from 1990-2000 and 2001- 

2011 did not show significant change regardless of extent of 

disease; localized CDS patients was 6.6 months (1990-2000) 

compared to 6 months (2001-2011). Conclusion: Over the last 

two decades, there has been an increased incidence in patients 

receiving CDS likely due to advancements in hepatic resection. 

The overall survival in the elderly population with CDS (75+) 

though remains poor throughout the last two decades regardless 

of the extent of the disease. Advanced age seems to be a 

poor prognostic factor even with CDS, suggesting future study 

in this age population. 

Disclosures: 

The following authors have nothing to disclose: George Cholankeril, Ponnandai 

Somasundar 

373 

Patients with alcohol compared to HCV-related hepatocellular 

carcinoma (HCC) have a reduced survival. 

Results of a Nationwide study. 

Charlotte E. Costentin 1 , Nathalie GOUTTE 2 , Philippe Sogni 3 , 

Bruno Falissard 4 , Noelle Bendersky 5 , Olivier Farges 2 ; 1 Hepatology, 

Hôpital Henri Mondor, Creteil, France; 2 Hepatology – Hepato-biliary 

surgery, Beaujon Hospital, Clichy, France; 3 Hepatology, 

Cochin Hospital, Paris, France; 4 biostatistique, Paris-Sud University, 

INSERM, Paris, France; 5 Medical Informatics, Beaujon Hospital, 

Clichy, France 

Introduction: Data on alcohol-related HCC are scarce. The 

aim of this study was to describe the incidence, management 

and prognosis of alcohol compared to HCV-related HCC, at a 

national level. Patients and Methods: French healthcare databases 

were screened to identify all adult patients who were 

hospitalized between 2007 and 2012 with a diagnosis of 

HCC. Incident cases of HCC were selected by defining the 

entry point as a first admission in 2009. Demographic data, 

associated conditions, underlying liver disease and etiology, 

as well as the type (teaching, general, private, charitable), 

location, and annual HCC-caseload of the hospitals where 

patients were first managed (1st tertile 45), were retrieved. Treatments were divided into 

curative (liver transplantation, resection, radiofrequency) or 

not. Survival of incident-cases was computed from the time of 

diagnosis and adjusted for potential confounding variables. 

In order to compare two homogeneous populations of alcohol 

and HCV-related HCC, patients with unknown etiology of 

liver disease, mixed etiologies or HBV were excluded, Results: 

Between 2009 and 2012, the study population included 

13,942 (83.4%) incident-cases of alcohol and 2,774 (16.6%) 

HCV-related HCC. The incidence of alcohol- and HCV-related 

HCC varied up to 2 fold from one mainland region to another, 

and in some regions, alcohol accounted for up to 96% of HCC. 

Alcohol- compared to HCV-related HCC occurred more often 

in men (89.2% vs 63.7% p


ing program were achieved as all of the patients completed 

the follow-up till dead. Forty patients were identified as HCC 

during the 10 years follow-up. The totally 1-, 3-, 5-, and 10-year 

cumulative HCC incidence rate were 1.6% (n=6), 4.6%(n=18), 

7.1%(n=27) and 10.5% (n=40) respectively. Among 40 HCC 

patients, thirty-eight (95%) were identified as asymptomatic 

subclinical early HCC with tumor size less than 3.0cm, and 

without portal vein infiltration or metastasis. Thirteen patients 

had the chance of surgical operation, and 12 (92.3%) patients 

survived with no recurrence. Twenty-five patients received 

radiofrequency ablation or other palliative therapy because of 

liver function reserve limitation. Though totally sixteen patients 

died, only 3 (18.8%) patients died of HCC(two were late stage 

HCC with metastasis, one was liver failure after operation), 

other 13(81.2%) patients died of concomitant liver decompensation 

complications. The median survival time is 3 years after 

identification of HCC. The totally 1-, 3-, and 5-year survival 

rate of HCC patients were 97.5%, 91.2%, 67.7% respectively. 

Conclusion: This 10 years regular integrated follow-up experience 

indicates that HCC surveillance using ultrasonography 

at six month intervals really performs good with early HCC 

detection rate of 95%. 

Disclosures: 

Jidong Jia - Consulting: BMS, MSD, Roche 

The following authors have nothing to disclose: Xiaoning Wu, Xiaoming Wang, 

yameng sun, Yuanyuan Kong, Yu Wang, Xinyan Zhao, Qianyi Wang, Weijia 

Duan, Hong You, Xiaojuan Ou 

375 

Integrator complex subunit 6 (INTS6) is a prognostic 

marker for hepatocellular carcinoma 

Minqiang Lu 1 , Ka Yin LUI 1 , Rongdang Fu 1 , Haoran Peng 1,2 , Huan 

Chen 1 ; 1 Hepatic Surgery, The Third Affiliated Hospital of Sun Yatsen 

University, Guangzhou, China; 2 Division of Gastroenterology 

and Hepatology, The Johns Hopkins Hospital, Baltimore, MD 

Purpose: Integrator complex subunit 6 (INTS6), previously 

known as the gene encoding deleted in cancer cells 1 (DICE1) 

was found to play a tumor suppressive role in certain types of 

solid tumors just like prostate cancer and cervical cancer. However, 

the clinical characteristic of INTS6 hasn’t been reported 

by far. In this study, we wanted to investigate the expression of 

INTS6 in hepatocellular carcinoma (HCC) and evaluated the 

clinical characteristic and prognosis in HCC patients. Material 

and methods: (1) Firstly, the expression level of INTS6 mRNA 

were measured in a cohort of 50 HCC tissues and adjacent 

normal liver tissues by using the qRT-PCR; (2) Secondly, the 

expression level of INTS6 protein were detected in 20 paired 

HCC and corresponding adjacent normal tissue by using western 

blot; (3) At last, Immunohistochemistry was performed on 

70 archived paraffin-embedded HCC samples. Results: qRT- 

PCR and western blot analyses showed the INTS6 mRNA and 

protein expression was significantly down-regulated in tumor 

tissues compared to adjacent normal liver tissues (p


min ratio and NLR demonstrated a prognostic value in HCC 

patients treated with sorafenib in a western collective. Inflammation 

based scores have the potential of clinical utility to identify 

patients who are likely not to derive significant benefit from 

a systemic therapy. References J Budczies, F Klauschen, BV 

Sinn, B Gyrffy, WD Schmitt, S Darb-Esfahani, C Denkert. Cutoff 

Finder: a comprehensive and straightforward Web application 

enabling rapid biomarker cutoff optimization.. PLoS One 7, 

e51862 (). 

Disclosures: 

Martin F. Sprinzl - Grant/Research Support: GILEAD 

The following authors have nothing to disclose: Arndt J. Weinmann, Sandra 

Koch, Peter R. Galle, Marcus Wörns 

377 

Development of a model predicting survival of patients 

with recurrent or progressive hepatocellular carcinoma: 

MOREPROH score 

Sang Il Choi 1 , Ami Yu 2 , Bo Hyun Kim 1 , Eun Jeong Ko 1 , Sun Seob 

Park 1 , Byung Ho Nam 2,3 , Joong-Won Park 1,3 ; 1 Center for Liver 

Cancer, National Cancer Center, Korea, Goyang-si, Korea (the 

Republic of); 2 Biometric Research Branch, National Cancer Center, 

Korea, Goyang-si, Korea (the Republic of); 3 Department of 

Cancer Control and Policy, Graduate School of Cancer Science 

and Policy, National Cancer Center, Korea, Goyang-si, Korea (the 

Republic of) 

Introduction: Most prognostic models developed for hepatocellular 

carcinoma (HCC) are based on data collected at the time 

of initial diagnosis. However, HCC frequently recur or progress 

after the initial treatment and the patient’s tumor burden, 

underlying liver function, and performance status are certain to 

change over time, affecting the prognosis of the patient. Therefore, 

we aimed to develop a model to predict the survival of 

patients at the time of disease recurrence or progression after 

the initial treatment. Methods: 1,972 patients in the cohort that 

consist of patients who were newly diagnosed as HCC at the 

National Cancer Center, Korea between January 2004 and 

December 2009, were followed till May 2014, and 1,301 

patients who had disease recurrence or progression after the 

initial treatment were identified. They were randomly assigned 

into the development cohort (75%, n = 976) and the validation 

cohort (25%, n = 325). Variables that significantly influence the 

survival of patients were sorted out and a survival prediction 

model for recurrent or progressive HCC patients was formulated 

using the multivariate Cox proportional hazards model. 

Performance of the model was evaluated using the c-statistics 

for discrimination ability and Hosmer-Lemeshow χ2 statistics 

for calibration ability on the both development cohort and validation 

cohort. Results: According to the multivariate analysis, 

age, serum albumin level, Model For End-Stage Liver Disease 

(MELD) score, tumor factors (such as number of nodules, size of 

the largest nodule, vascular invasion, and extrahepatic metastasis), 

serum alpha-fetoprotein (AFP) level, presence of ascites, 

initial treatment modality, and the best response following the 

initial treatment were independent prognostic factors for overall 

survival. Using these variables, survival prediction model was 

developed: MOREPROH score (Model predicting survival of 

patients with Recurrent or Progressive HCC) MOREPROH score 

was tested on the development cohort and the predefined validation 

cohort. The respective c-statistics and χ2 statistics of 

this novel model for the validation cohort were 0.836 (95% 

confidence interval [CI] 0.806-0.865) and 6.339(p = 0.706); 

0.808 (95% CI 0.781-0.834) and 4.408(p = 0.883); and 

0.803 (95% CI 0.777-0.829) and 10.960(p = 0.278) for 1-, 

3-, and 5-year survival, respectively. Conclusion: A new model, 

MOREPROH to predict survival for patients with recurrent or 

progressive HCC using only objective variables was developed 

and validated for the first time. This model may be useful for 

planning of subsequent treatment strategy. 

Disclosures: 

The following authors have nothing to disclose: Sang Il Choi, Ami Yu, Bo Hyun 

Kim, Eun Jeong Ko, Sun Seob Park, Byung Ho Nam, Joong-Won Park 

378 

Undetected asymptomatic alcoholic cirrhosis underlies 

the diagnosis of hepatocellular carcinoma (HCC) out of 

surveillance programs in Spain. Analysis of 720 cases 

in 74 centers. 

Carlos Rodriguez-Lope 1 , Maria Elisa Reig 2 , Ana Matilla 3 , Maria 

Teresa Ferrer 4 , Eva Dueñas 5 , Beatriz Minguez 6 , Javier F. Castroagudin 

7 , Inmaculada Ortiz 8 , Sonia Pascual 9 , Jose Luis Lledo 10 , 

Adolfo Gallego 11 , Juan I. Arenas 12 , Carles Aracil 13 , Montserrat 

Forne 14 , Carolina Muñoz 15 , Fernando Pons 16 , Margarita 

Sala 17 , Mercedes Iñarrairaegui 18 , Marta Martin-llahi 19 , Victoria 

Andreu 20 , Carmen Garre 21 , Paloma Rendon 22 , Javier Fuentes 24 , 

Javier Crespo 1 , Manuel Rodríguez 23 , Jordi Bruix 2 , Maria Varela 23 ; 

1 Liver Unit, Hospital Universitario Marqués de Valdecilla. IDIVAL, 

Santander, Spain; 2 Liver Unit. BCLC group., Hospital Clinic de 

Barcelona, Barcelona, Spain; 3 Gastroenterology and Hepatology, 

H.G.U. Gregorio Marañon, Madrid, Spain; 4 Gastroenterology 

and hepatology, H. Virgen del Rocío, Sevilla, Spain; 5 Gastroenterology 

and Hepatology, H. de Bellvitge, Hospitalet de Llobregat, 

Spain; 6 Liver Unit, H. Vall d’Hebrón, Barcelona, Spain; 7 H. Clínico 

Universitario de Santiago, Santiago de Compostela, Spain; 8 H. 

Universitario Doctor Peset, Valencia, Spain; 9 H.G.U. de Alicante, 

Alicante, Spain; 10 H. U. Ramón y Cajal, Madrid, Spain; 11 H. de la 

Santa Creu i Sant Pau, Barcelona, Spain; 12 H. Donostia, Donostia, 

Spain; 13 H. U. Arnau de Vilanova, Lleida, Spain; 14 H. U. Mutua 

de Terrassa, Terrassa, Spain; 15 H. U. 12 de Octubre, Madrid, 

Spain; 16 H. U. Puerta de Hierro, Madrid, Spain; 17 H. U. Germans 

Trias i Pujol, Badalona, Spain; 18 Clinica Universitaria de Navarra, 

Pamplona, Spain; 19 H. Moises Broggi, Sant Joan Despi, Spain; 

20 H. de Viladecans, Barcelona, Spain; 21 H. U. Virgen de la Arrixaca, 

Murcia, Spain; 22 H. Puerta del Mar, Cadiz, Spain; 23 Liver 

Unit, H.U. Central de Asturias, Oviedo, Spain; 24 H.U. Miguel Servet, 

Zaragoza, Spain 

Introduction. Between Oct’08-Jan’09 we performed a survey 

of HCC with 705 cases in 62 centers in Spain[Med Clin 

(Barc).2010;134(13):569-7].In that study 53% of patients 

were diagnosed out of surveillance with more advanced HCC 

and less radical therapies applied. We have performed a new 

study to analyze the reasons of not performing surveillance. 

Methods. We invited previous participants extending the invitation 

to any center in Spain via Spanish Association for the 

Study of the Liver website. Investigators prospectively registered 

demographic, clinical and tumor characteristics, first treatment 

and eligibility for liver transplantation (OLT). Results. Between 

Oct’14-Jan’15, 74 centers registered 720 new cases of primary 

liver cancer: HCC (n=686), intrahepatic cholangiocarcinoma 

(ICC) (n=29), mixed HCC-ICC (n=2), others (n=3). 

HCC characteristics were: men 82%; age 67 years; cirrhosis 

87%. Main etiologies: alcohol 35%, HCV 30%, HCV + alcohol 

15%, NAFLD 6%; Child-Pugh 6 (5-13); number of nodules 1 

(1-20); size of the largest 3cm (1–24); vascular invasion 18%; 

extrahepatic disease 8%. BCLC stages: 0: 11%, A: 43%, B: 

20%, C: 16% and D: 11%. Main treatments: percutaneous 

ablation (22%), TACE (23%), resection (11%) and sorafenib 

(11%). 10% (n=67) of patients were evaluated for OLT. 53% 

(n=316) were diagnosed outside surveillance: 269 (76%) by 

chance or at the same time that their liver disease; 63 (18%)


were not adherent to surveillance; and 21 (6%) had chronic 

liver disease without a clear cirrhosis. Patients diagnosed out 

of surveillance had more advanced BCLC (p


381 

Prediction of Hepatocellular Carcinoma Risk in Patients 

with Cirrhosis: Validation of ADRESS–HCC Model 

Ju Dong Yang, Hager Ahmed Mohammed, Jonggi Choi, Gregory 

J. Gores, Lewis R. Roberts, W. Ray Kim; Division of Gastroenterology 

and Hepatology, Mayo Clinic College of Medicine, Rochester, 

MN 

Background/Aims: ADRESS-HCC is a model to predict the risk 

of hepatocellular carcinoma (HCC) in patients with cirrhosis 

[Cancer 2014;3485]. It incorporates age, diabetes, race, 

etiology and severity of cirrhosis, and sex. In this study, we 

validate the model in an independent cohort of patients. Methods: 

Of all cirrhotic patients seen at Mayo Clinic Rochester 

between January 2006 and December 2011, those who had 

liver images (US, CT, or MRI) at baseline and at follow up more 

than 6 months from baseline were included. The ADRESS risk 

score was calculated by the following: score= [age]*0.0532+ 

[1 for diabetes]*0.2135 + [1 for nonwhite/Hispanic]*0.2058 

+ [1 for metabolic/alcohol etiology]*0.3509 + [1 for viral 

etiology]*1.246 + [1 for male]*0.5114 + [Child-Turcotte-Pugh 

(CTP) score]*0.1170. Observed incidence of HCC was compared 

to the model’s prediction. Results: A total of 739 cirrhotic 

patients met the eligibility criteria. The mean age was 57.4 

years, with 59% men, 90% non-Hispanic White and 34% diabetic. 

The etiology of liver disease was alcoholic in 32%, viral 

in 23% and metabolic in 23%. The mean CTP score was 8. 

After a median follow up of 38 months, 69 (9%) patients developed 

HCC. The median ADRESS score was 5.01 (interquartile 

range: 4.44-5.46). Figure divides patients into 3 groups by 

the ADRESS score quartiles: the top (high risk) quartile (hazard 

ratio [HR]=6.8; 95%CI=2.9-20.2) and the middle two quartiles 

(HR=3.2; 95%CI=1.4-9.4) had a significantly higher incidence 

of HCC compare to the lowest risk quartile. When the surveillance 

threshold (ADRESS=4.67 with predicted annual incidence=1.5%) 

was applied, 0.97% of patients developed HCC 

in the first 18 months. Conclusions: Based on an independent 

cohort of cirrhotic patients, we validate ADRESS-HCC model as 

a useful tool for predicting the risk of HCC among patients with 

cirrhosis and identifying candidates for surveillance. 

Disclosures: 



The following authors have nothing to disclose: Beatriz Minguez, Ayse Aytaman, 

Meritxell Ventura-Cots, Maaz B. Badshah, Caitlin C. Citti, Heather L. Platt, Ting-Yi 

Chen, Luciana Kikuchi, Sonja Marcus, Michael Yin, Judith Aberg, Myron E. 

Schwartz, Norbert Bräu 

Disclosures: 







The following authors have nothing to disclose: Ju Dong Yang, Hager Ahmed 

Mohammed, Jonggi Choi 

382 

The life time frequency and interval of treatments define 

the prognosis of hepatocellular carcinoma after radical 

radiofrequency ablation therapy 

Takamasa Ohki, Yuki Hayata, Satoshi Kawamura, Daisaku Ito, 

Koki Kawanishi, Kentaro Kojima, Michiharu Seki, Nobuo Toda, 

Kazumi Tagawa; Gastroenterology, Mitsui Memorial Hospital, 

Tokyo, Japan 

Backgrounds and Aims: Hepatocellular carcinoma (HCC) 

frequently recurs even after curative radiofrequency ablation 

(RFA) therapy and the recurrence rate is approximately 50% 

within 3 years. Thus, the treatment strategy after HCC recurrence 

is very important to achieve a long term survival. We 

analyzed the life time frequency and interval of treatments to 

evaluate their impacts on overall survival (OS). Patients and 

methods: In this retrospective study, we enrolled 305 naïve 

HCC patients who were curatively treated with RFA between 

January 1 2000 and December 31 2010. The last follow-up 

date was December 31 2014. The rate of life time frequency of 

treatments per year (LFT rate) was calculated as total numbers 

of treatments divided by observational year. The total numbers 

of treatments were defined as sum of frequency of RFA and 

trans-catheter chemo-embolization or infusion (TACE or TAI) 

times. Patients were stratified into four groups to compare the 

OS according to LFT rate: group A; LFT rate < 0.5 N = 53, 

group B; 0.5 ≤ LFT rate < 1.0 N = 69, group C; 1.0 ≤ LFT rate 

< 2.0 N = 103, and group D; 2.0 ≤ LFT rate N = 80. Results: 

During the mean follow-up of 4.7 years, the life time frequency 

of treatments was 5.2 times on average. The frequency of each 

treatment modality was as follows: RFA 2.8 and TACE or TAI 

2.3 times. One-hundred fifty-eight patients died during the follow-up 

period, showing cumulative survival rates at 3, 5 and


10 years of 100%, 96.0%, 81.2% in group A, 95.3%, 77.6% 

and 45.5% in group B, 70.1%, 41.7% and 8.7% in group C, 

and 50.4%, 22.2% and 0% in group D, respectively (P < 0.01 

by the log-rank test). Adjusting for significant factors in univariate 

analysis, multivariate analysis indicated that frequency of 

receiving RFA (HR: 0.60 pre 1 time, P < 0.01), frequency of 

receiving TACE or TAI (HR: 0.88 per 1 time, P < 0.01) and 

LFT rate over 1.0 (HR: 29.4, P < 0.01) as independent factors 

which affected on OS. First recurrence, tumor sizes, and tumor 

numbers were not significant risk factors in the univariate analysis. 

Conclusion: Appropriate treatment at the time of each HCC 

recurrence remarkably prolonged OS even if HCC patients had 

multiple relapse. Repeat RFA and TACE or TAI were promising 

strategy to achieve a long term survival. However, LFT rate 

over 1.0 which indicated high rate of HCC recurrence was an 

independent factor of poor prognosis suggesting the malignant 

potential of HCC. 

Disclosures: 

Takamasa Ohki - Speaking and Teaching: Otsuka Pharmaceutical Co. Ltd. 

The following authors have nothing to disclose: Yuki Hayata, Satoshi Kawamura, 

Daisaku Ito, Koki Kawanishi, Kentaro Kojima, Michiharu Seki, Nobuo Toda, 

Kazumi Tagawa 

383 

Efficacy and safety of miriplatin versus epirubicin in 

transcatheter arterial chemoembolization therapy for 

advanced hepatocellular carcinoma: a randomized controlled 

trial 

Tatehiro Kagawa 1 , Shunji Hirose 1 , Yoshitaka Arase 1 , Kota 

Tsuruya 1 , Kazuya Anzai 1 , Sei-ichiro Kojima 1 , Koichi Shiraishi 1 , 

Masako Shomura 3 , Jun Koizumi 2 , Tetsuya Mine 1 ; 1 Gastroenterology, 

Tokai University, Isehara, Japan; 2 Radiology, Tokai University 

School of Medicine, Isehara, Japan; 3 Nursing, Tokai University 

School of Medicine, Isehara, Japan 

Purpose: Miriplatin, a hydrophobic platinum-based anticancer 

agent, is used for transcatheter arterial chemoembolization 

(TACE), but its efficacy and safety are still unclear. To clarify 

the efficacy and safety of TACE with miriplatin in the treatment 

of patients with advanced hepatocellular carcinoma (HCC), 

we performed a randomized trial to compare miriplatin with 

epirubicin. Patients and Methods: We randomly assigned 

patients with advanced HCC to receive TACE with miriplatin 

or TACE with epirubicin after stratification by the tumor 

diameter (3 cm). Enrollment criteria were (1) presence of HCC 

with contrast-enhancement in the arterial phase of dynamic CT 

scan, (2) no indication of surgical resection or local therapy 

including radiofrequency ablation and percutaneous ethanol 

injection, (3) no previous treatment for the target HCC, (4) no 

extrahepatic metastasis, (5) no tumor thrombus in the portal 

or hepatic veins, (6) Child-Pugh class A or B, (7) ECOG PS 

score of 2 or less, and (8) serum bilirubin < 3 mg/dl, creatinine 

< 1.5 mg/dl, white blood cell count ≥ 3000/mm 3 , 

platelet count ≥ 50000/mm 3 , and hemoglobin concentration 

≥ 9.5 g/dl. Dynamic CT was performed within 3 months after 

TACE and the efficacy was blindly evaluated by two radiologists 

according to the modified Response Evaluation Criteria in 

Solid Tumors (mRECIST). Overall survival and recurrence-free 

survival were compared by log-rank test. The treatment modality 

for the recurrence was decided by the primary doctors. 

This study was approved by the institutional ethics committee 

(UMIN000003987) and written informed consent was 

obtained from each patient. Results: 23 and 24 patients were 

allocated to the miriplatin (M) group and the epirubicin (E) 

group, respectively. No significant difference was observed in 

age gender, etiology, tumor size, Child-Pugh sore, and clinical 

stage between two groups. The rate of complete response in 

the M group was significantly lower than in the E group (33% 

vs 79%, P < 0.01). Recurrence-free survival was significantly 

shorter in the M group than in the E group (P < 0.05); 1-yr 

recurrence-free survival rate was 18% in the M group and 33% 

in the E group. OS was not significantly different between two 

groups; 1-yr and 3-yr OS were 95% and 58%, respectively, in 

the M group and 88% and 79%, respectively, in the E group. 

As adverse effects grade 2 fever was observed with similar 

frequency; 19% in the M group and 32% in the E group. Conclusions: 

This randomized trial demonstrated that the efficacy 

of the miriplatin-TACE was inferior to the conventional epirubicin-TACE, 

although miriplatin was as safe as epirubicin. 

Disclosures: 

The following authors have nothing to disclose: Tatehiro Kagawa, Shunji Hirose, 

Yoshitaka Arase, Kota Tsuruya, Kazuya Anzai, Sei-ichiro Kojima, Koichi Shiraishi, 

Masako Shomura, Jun Koizumi, Tetsuya Mine 

384 

Identification of candidate extracellular non-coding RNA 

biomarkers for Hepatocellular Carcinoma 

Irene K. Yan, Tushar Patel; Mayo Clinic, Jacksonville, FL 

Hepatocellular carcinoma (HCC) has a poor prognosis. Early 

detection may improve outcomes by detecting the cancer at a 

stage when curative procedures may be possible. Our overall 

goals are to evaluate the utility of extracellular non-coding 

RNA as biomarkers for the early detection of HCC. Towards 

this goal, we sought to identify non-coding extracellular RNA 

(exRNA) that are associated with HCC cells, and to develop 

assays for their detection. METHODS: A systematic profiling 

approach evaluated non-coding RNA that are (1) associated 

with malignant HCC cells and (2) expressed within extracellular 

vesicles (EV) released from these cells. qPCR, Nanostring 

and droplet digital PCR were used for the detection of 

exRNA from culture supernatants or from serum of patients with 

HCC. Profiling was performed on RNA from cells and exRNA 

released by these cells for non-malignant (HH and THLE-2), and 

malignant HCC cells (Hep3B, HepG2, PLC/PRF/5, SNU-182, 

SNU-398). MicroRNA expression was assessed using qPCR 

panels from Exiqon to screen 752 miRNAs, and expression of 

90 selected lncRNAs was assessed using lncRNA qPCR assays. 

The expression of 800 miRNAs and 33 lncRNAs was analyzed 

in selected samples using Nanostring. Assay precision 

and reproducibility were individually determined for selected 

exRNA candidate biomarkers. RESULTS: HCC cells released 

a larger amount of extracellular RNA compared to normal 

hepatocytes (average of 1.23% vs. 0.55% of total donor cell 

RNA). Compared to exRNA from normal hepatocytes, qPCR 

analyses identified 14 lncRNAs and 33 miRNAs that were 

increased in exRNA from HCC cells. In addition, Nanostring 

analysis identified three miRNAs (miR-25, miR-302d, and miR- 

612) in exRNA preparations from all HCC cells, and from 

serum obtained from patients with HCC. These three microR- 

NAs were also detected using droplet digital PCR but not using 

qPCR. Droplet digital PCR assays for long non-coding RNA 

GAS5 and H19 in exRNA had a CV of 10.6% and 7.1% for 

precision, and a CV of 11% and 3% for reproducibility respectively. 

Digital PCR assays and nanostring were more sensitive 

than qPCR assays for the detection of low abundance exRNA 

in serum. SUMMARY and CONCLUSIONS: Non-coding RNAs 

that are selectively enriched within exRNA from HCC cells can 

be detected in the circulation. Using a novel biomarker discovery 

platform based on exRNA release, we have identified 

several candidate microRNA and long non-coding RNA biomarkers 

for HCC. Sensitive assay platforms have been devel-


oped for use in studies to validate the utility of these exRNA as 

diagnostic markers for HCC. 

Disclosures: 

The following authors have nothing to disclose: Irene K. Yan, Tushar Patel 

385 

The radiofrequency ablation is eligible treatment for 

the case of hepatocellular carcinoma more than 3 cm 

in diameter with Child-Pugh class A or des-gamma-carboxy 

prothrombin level less than 200 mAU/mL: Analysis 

of 1815 cases in a single center for fourteen-year 

experience 

Takashi Tanaka, Akira Anan, Daisuke Morihara, Kazuhide 

Takata, Keiji Yokoyama, Yasuaki Takeyama, Makoto Irie, Satoshi 

Shakado, Tetsuro Sohda, Shotaro Sakisaka; Fukuoka university 

school of medicine, Hepatology, Fukuoka, Japan 

Background and Aims: Most of the hepatocellular carcinoma 

(HCC) more than 3 cm in diameter is categorized into intermediate 

stage of Barcelona Clinic Liver Cancer (BCLC) staging 

system, and first line treatment in the intermediate stage is recommended 

transcatheter arterial chemoembolization (TACE). 

Although general indication of radiofrequency ablation (RFA) 

treatment for HCC is defined as tumor size less than 3 cm in 

diameter, some previous trials described RFA for HCC more 

than 3 cm in diameter. The aim of this study was to analyze the 

long term outcome and prognostic factors in the patients who 

had received RFA treatment for HCC more than 3 cm in diameter. 

Methods: A total of 1815 cases with liver tumors, from 

April 2000 to September 2014 were received RFA treatment 

in Fukuoka University Hospital. Among them, 94 cases were 

enrolled in this study, which was defined as patients with single 

or multinodular HCC more than 3 cm in diameter. Overall survival 

and recurrence rates were analyzed by the Kaplan-Meier 

method and risk factors on prognosis was assessed by multivariate 

Cox regression hazard model. Results: The median age of 

the study population (men 62 and women 32) was 71 (range: 

48-88) years old. Patient’s Child-Pugh class was in A (n=72) 

and B (n=22) and 77 patients related with viral hepatitis (HBV: 

n=4, HCV: n=73). The median tumor size was 3.5 cm in diameter 

(range: 3.1-6.2 cm), and the number of tumor was single: 

n=51, double: n=20, triple: n=6, more than four: n=7. We 

declined “up to 6” group who have the number of the tumor 

plus maximal size (cm) less than six, 66 patients were within 

up to 6 group. The median serum level of alpha-fetoprotein 

(AFP) and des-gamma-carboxy prothrombin (DCP) level was 35 

(range: 2-3061) and 99 (range: 8-12502) respectively. Cumulative 

overall survival rates at 1-, 3-, 5-, 10-years were 93.4%, 

62.7%, 38.4%, and 12.7%, respectively. Cumulative disease 

free survival rate at 1-, 3-, 5-, 10-years were 51.4%, 19.4%, 

15.3%, and 4.6%, respectively. Multivariate analysis revealed 

that factors associated with survival were tumor recurrence free, 

(HR=3.428; 95%CI 1.407-11.421, p=0.0044), Child-Pugh 

class A (HR=2.136; 95%CI 1.233-3.633, p=0.0084), and 

DCP levels less than 200 mAU/mL (HR=1.957; 95%CI 1.198- 

3.159, p=0.0078). Multivariate analysis disclosed that without 

up to 6 (HR=1.714; 95%CI 1.066-2.708, p=0.027) predicted 

higher incidence of developing recurrence. Conclusion: From 

our over 1800 RFA experiences in fourteen years, we revealed 

that radiofrequency ablation is eligible treatment for the case 

of hepatocellular carcinoma more than 3 cm in diameter with 

Child-Pugh class A or DCP level less than 200 mAU/ml. 

Disclosures: 

The following authors have nothing to disclose: Takashi Tanaka, Akira Anan, Daisuke 

Morihara, Kazuhide Takata, Keiji Yokoyama, Yasuaki Takeyama, Makoto 

Irie, Satoshi Shakado, Tetsuro Sohda, Shotaro Sakisaka 

386 

Racial Differences in the Presentation and the Incidence 

of Hepatocellular Carcinoma (HCC) in a Large Cohort of 

Patients with Cirrhosis in the United States 

Long H. Nguyen 1 , Changqing Zhao 2,5 , Nghia H. Nguyen 3,2 , 

Joseph K. Hoang 2 , Michael D. Nguyen 2 , Richard H. Le 2 , Peter T. 

Nguyen 2,6 , Derek Lin 1 , Vinh D. Vu 2 , Lee Ann Yasukawa 4 , Susan C. 

Weber 4 , Mindie H. Nguyen 2 ; 1 Department of Medicine, Stanford 

University Medical Center, Palo Alto, CA; 2 Division of Gastroenterology 

and Hepatology, Stanford University Medical Center, Palo 

Alto, CA; 3 Department of Medicine, University of California, San 

Diego, San Diego, CA; 4 Center for Clinical Informatics, Stanford 

University School of Medicine, Palo Alto, CA; 5 Department of Cirrhosis, 

Institute of Liver Disease, Shuguang Hospital, Shanghai 

University of T.C.M., Shanghai, China; 6 School of Medicine, University 

of Texas Medical Branch, Galveston, Galveston, TX 

Purpose: Cirrhosis regardless of etiology is a significant HCC 

risk factor. The purpose of our study is to clarify the largely 

unknown role of ethnicity in the development of HCC in patients 

with cirrhosis. Methods: We conducted a retrospective cohort 

study of 3208 consecutive white (n=1,692), Hispanic (n=772), 

Asian (n=614), and black (n=130) patients with cirrhosis seen 

at a single university tertiary care center from 2005-2010. Our 

primary endpoint was HCC diagnosis by histology or non-invasive 

criteria. Results: Among ethnic groups, patients were 

similarly aged (54-59 years), mostly male (61-64%), and with 

similar baseline MELD scores (17-23), while black patients had 

much shorter duration of follow up (22 vs. 31-40 months). At 

presentation, the highest HCC prevalence was seen in Asian 

(11%) then Hispanic (5%), white (4%), and black patients (4%, 

p


patients were less likely to meet Milan criteria for liver transplantation 

(LT) compared to HCV and nonviral patients: 34%, 

50%, 39%, P < 0.0001). HBV patients were more likely than 

HCV and non-viral patients to undergo resection (26%, 9%, 

and 13%, respectively; P


older (mean ages 60 ± 13 vs. 55 ± 10, p


RFA vs. 52.5% in HR (p=0.677). On 3cm < size ≤ 5cm, cumulative 

DFS were 29.6% in TACE and RFA vs. 40.7% in HR 

(p=0.513). Most common complication on both group was 

elevation of AST/ALT (grade 2), most severe complication were 

happened 3 cases of HR group. One is severely elevated bilirubin 

without encephalopathy; the others were death due to 

renal failure or sepsis. In TACE with RFA group, liver segmental 

infarction were happened in 2 cases. Conclusions: Combined 

TACE and RFA could be alternative choice of initial treatment 

in early HCC patients. 

Disclosures: 

Kwan Soo Byun - Grant/Research Support: Gilead, BMS 

The following authors have nothing to disclose: Young Kul Jung, Sang Jun Suh, 

Hyung Joon Yim, Oh Sang Kwon, Yun Soo Kim, Duck Joo Choi, Ju Hyun Kim, Ji 

Hoon Kim, Jong Eun Yeon, Yeon Seok Seo, Soon Ho Um, Ho Sang Ryu 

392 

Myeloid-derived suppressor cells correlate with patient 

outcomes in hepatic arterial infusion chemotherapy for 


Eishiro Mizukoshi, Tatsuya Yamashita, Kuniaki Arai, Takeshi Terashima, 

Masaaki Kitahara, Hidetoshi Nakagawa, Noriho Iida, 

Kazumi Fushimi, Shuichi Kaneko; Kanazawa Univ, Kanazawa, 

Japan 

Background: The modulation of tumor-associated host immune 

responses after cancer treatments has recently been reported. 

For example, hepatocellular carcinoma (HCC) treatments with 

radiofrequency ablation or transarterial chemoembolization 

were found to increase the frequency of tumor-associated antigen 

(TAA)-specific T cells due to the creation of an antigen 

source by the destruction of tumor cells. In addition, recent 

studies demonstrated that beyond their direct cytotoxic effects 

on cancer cells, several conventional chemotherapeutic drugs 

promoted the elimination or inactivation of regulatory T cells 

(Tregs) or myeloid-derived suppressor cells (MDSCs), resulting 

in enhanced host antitumor immunity. Hepatic arterial infusion 

chemotherapy (HAIC) has been employed as an alternative 

therapy to sorafenib for the patients with advanced HCC in 

Japan and other Asian countries. In the present study, we 

performed a comparative analysis of various immune cell 

responses including TAA-specific T cells, Tregs and MDSCs in 

advanced HCC patients treated with HAIC, and analyzed the 

relationship between these immune cell responses and patient 

prognosis. Methods: Thirty-six HLA-A24-positive patients who 

had clinically diagnosed advanced HCC were examined in the 

present study. After being diagnosed, all patients were treated 

with HAIC using interferon (IFN)/ 5-fluorouracil (FU) or IFN/ 

FU + cisplatin. Interferon gamma ELISPOT assays were performed 

to examine the frequency of TAA-specific T cells using 

11 peptides derived from 5 TAAs. The frequencies of Tregs and 

MDSCs were examined by multicolor fluorescence-activated 

cell sorting analysis and analyzed the association with clinical 

factors and prognosis of patients. Results: The frequency of 

Tregs was significantly decreased after HAIC but no significant 

differences were observed in the frequency of TAA-specific T 

cells and MDSCs. An analysis of prognostic factors for overall 

survival identified diameter of the tumor (


tic thrombosis had better survival when treated with anticoagulant 

(log-rank test p=0.001). Conclusion: Portal vein thrombosis 

in HCC patients is non-malignant in half of the patients, calling 

for anticoagulant therapy. The prognosis of patients with neoplastic 

and non neoplastic thrombosis remain poor. 

Disclosures: 









The following authors have nothing to disclose: Angelo Sangiovanni, Cristina 

Bertelli, Sara Vavassori, Giuseppina Pisano, Massimo Iavarone, Silvia Fargion, 

Anna Ludovica Fracanzani 

394 

Preclinical characterization of a novel first in class 

GNS561: autophagy inhibitor therapeutic candidate for 

treatment of hepatocellular carcinoma 

Firas Bassissi 1 , Sonia Brun 1 , Jerome Courcambeck 1 , Gregory 

Nicolas 1 , Clarisse Dubray 1 , Antoine Beret 1 , Paul Castellani 2 , 

Xavier Adhoute 2 , Marc Bourlière 2 , Jean-Luc Raoul 3 , Philippe Halfon 

1 ; 1 Genoscience Pharma, Marseille, France; 2 Hepatology, 

Hopital Saint-Joseph, Marseille, France; 3 Oncology, Institut Paoli 

Calmettes, Marseille, France 

Background: In spite of successful approval and wide application 

of sorafenib, the prognosis for patients with advanced 

hepatocellular carcinoma (HCC) remains poor. Fortunately, 

there have been renewed and continued interests and active 

research in developing other molecularly targeted agents in 

HCC during the past few years. While there is early evidence 

of antitumor activity of several agents in phase I/II studies, 

phase III efforts with a few targeted agents have failed, highlighting 

the challenges in new drug development in HCC. For 

this reason, development of innovative drugs with original 

mechanism of action could improve treatment of HCC patients 

Material and methods: We screened a library of autophagy 

inhibitor compounds and identified a new drug GNS561 as 

compound that kills tumor cells in a panel of HCC cell lines 

and primary tumor. Drug tolerance and plasma and liver pharmacokinetic 

were evaluated after single and repeated dosing 

in mice and rat. Results: GNS561 demonstrates autophagy 

inhibition and apoptosis induction activities related to lysosome 

disruption. GNS 561 shows potent anti-proliferation activity 

when assayed against a panel of human tumor cell lines, notably 

against HCC cell lines (Mean EC50 2mM). In addition 

GNS561 demonstrates potent activity against a panel of HCC 

patient tumors even in those with sorafenib resistance (Mean 

EC50 3mM vs 11mM for sorafenib). GNS561 is highly selectively 

trapped in the liver, via uptake hepatocyte transporters. 

Plasma and liver PK in mice and rats after single and repeated 

doses confirm this selectivity (exposure ratio liver/plasma 

about 20 in mice) with a good oral bioavailability. Tolerance 

of single and repeated doses is excellent in both rats and mice 

Conclusions: Our results provide a rationale for testing autophagy 

flux disruption as a novel therapeutic strategy for HCC. 

GNS561 is a liver selective drug which offers great promise 

for HCC treatment of non-responder or nontolerant patients to 

sorafenib. This compound is selected as a drug candidate for 

future investigation in HCC clinical trials. 

Disclosures: 

Xavier Adhoute - Speaking and Teaching: bayer 






Squibb 

Jean-Luc Raoul - Advisory Committees or Review Panels: Bayer SP, BMS, BTG, 

Arqule; Speaking and Teaching: Bayer SP 

The following authors have nothing to disclose: Firas Bassissi, Sonia Brun, Jerome 

Courcambeck, Gregory Nicolas, Clarisse Dubray, Antoine Beret, Paul Castellani, 

Philippe Halfon 

395 

Survival of Patients with Advanced Hepatocellular 

Carcinoma Treated with Sorafenib: An Analysis of 

SEER-Medicare Database 

Neehar D. Parikh 1 , Vincent D. Marshall 2 , Hari Nathan 3 , Rajesh 

Balkrishnan 2 , Vahakn Shahinian 4 ; 1 Division of Gastroenterology, 

University of Michigan, Ann Arbor, MI; 2 College of Pharmacy, 

University of Michigan, Ann Arbor, MI; 3 Department of Surgery, 

University of Michigan, Ann Arbor, MI; 4 Kidney Epidemiology and 

Cost Center, University of Michigan, Ann Arbor, MI 

Sorafenib, a multikinase inhibitor, is the only chemotherapeutic 

approved for use in the US for the treatment of advanced HCC. 

We aimed to examine the outcomes of elderly patients with 

advanced HCC in the US. We performed a secondary analysis 

of continuously enrolled Medicare beneficiaries with HCC diagnoses 

from 2007-2009, based on SEER diagnosis codes. Our 

primary aim was to determine survival in patients treated with 

sorafenib and predictors of survival. We compared advanced 

stage patients with HCC (stage III/IV) who received sorafenib 

within 6 months of diagnosis to advanced stage patients with 

HCC who received no therapy (control.) We calculated the 

Charlson comorbidity index 12 months prior to diagnosis and 

an aggregate variable for decompensated cirrhosis (presence 

of esophageal varices or variceal banding, ascites or paracentesis, 

hepatic encephalopathy or use of neomycin, lactulose, 

or rifaximin.) We performed univariate (Kaplan Meier with 

log rank test) and multivariate (Cox regression) analyses for 

predictors of survival. The baseline characteristics of the patient 

are shown in the Table. Survivial was singificantly better in the 

sorafenib treated group (p


Disclosures: 

Vahakn Shahinian - Consulting: Amgen, Inc 

The following authors have nothing to disclose: Neehar D. Parikh, Vincent D. 

Marshall, Hari Nathan, Rajesh Balkrishnan 

396 

The Casein kinase II (CK2) inhibitor CX-4945 has 

an additive effect with gemcitabine and cisplatin on 

cholangiocarcinoma (CCA) cell lines mediated in part 

through PI3K/Akt inhibition 

Kais Zakharia 1 , Katsuyuki Miyabe 1 , Catherine D. Moser 1 , Sean 

O’Brien 3 , Mitesh J. Borad 2 , Lewis R. Roberts 1 ; 1 Division of Gastroenterology 

and Hepatology, Mayo Clinic College of Medicine, 

and Mayo Clinic Cancer Center, Rochester, MN; 2 Division of 

Hematology, Division of Oncology, Mayo Clinic, Scottsdale, AZ; 

3 Senhwa Biosciences, San Diego, CA 

Background: There is a need to develop effective therapies for 

CCA due to suboptimal outcomes with the current standard 

gemcitabine (Gem) and cisplatin (Cis) therapy. Casein kinase 

II (CK2) is a serine/threonine selective protein kinase that has 

been implicated in cancer progression, making it an attractive 

target for anticancer treatment. CX-4945, a first in class, orally 

available and highly selective inhibitor of CK2 has shown antitumor 

activity in other human cancers. In this study we investigated 

the anti-neoplastic effect of CX-4945 on 3 different 

CCA cell lines. Aims: To determine 1) whether CX-4945 has 

an anti-proliferative effect on CCA cells; 2) if CX-4945 induces 

apoptosis in CCA cells; 3) whether the anti-proliferative action 

of CX-4945 occurs through effects on PI3K/Akt signaling; and 

4) whether the effect of CX-4945 enhances that of Gem or Cis 

in vitro. Methods: CCK8 and colony formation assays were 

used to assess cell viability in cultures of HuCCT1 (intrahepatic 

CCA), EGI-1 (extrahepatic CCA), and a new primary patient 

derived CCA cell line LIV27 after treatment with CX-4945 

alone or in combination with Gem or Cis. The Caspase 3/7 

assay was used to assess the effect of CX-4945 on CCA apoptosis. 

The half maximal inhibitory concentrations (IC50) were 

calculated using the Chou-Talalay technique. Western immunoblotting 

was used to assess the effect of CX-4945 on Akt 

pathway signaling in CCA cells. Results: CX-4945 significantly 

decreased cell viability of HuCCT1, EGI-1 and LIV27 in a timeand 

dose-dependent manner (IC50 of 7.3, 9.5, and 9.4 μM 

respectively at 72 hours). CX-4945 inhibited colony formation 

in HuCCT1 and EGI-1 (IC50 of 3.8 and 1.6 μM respectively). 

CX-4945 did not significantly increase Caspase 3/7 activity 

in CCA cells until higher doses (15 and 20 μM) were used. 

CX-4945 inhibited Akt phosphorylation (at Serine473 and 

Serine129) and PTEN phosphorylation, suggesting that it acts 

through the PI3K/Akt pathway. Addition of CX-4945 to Cis 

or Gem decreased viability of the HuCCT1, EGI-1 and LIV27 

cells by 15-40% (depending on the dose and the cell line) 

with a combination index (CI) of approximately 1, indicating 

an additive effect. Addition of CX-4945 to both Gem and Cis 

together (1/4 IC50s) decreased viability of HuCCT1 cells by 

10-30%, also with a CI around 1. Conclusion: CX-4945 has an 

anti-proliferative effect on CCA cell lines. Only higher doses of 

CX-4945 increased Caspase 3/7 activity. The anti-proliferative 

effect of CX-4945 correlates with effects on PI3K/Akt signaling. 

CX-4945 enhanced the anti-proliferative effect of both Cis and 

Gem in vitro. An animal study is being performed to confirm 

these findings in vivo. 

Disclosures: 

Sean O’Brien - Employment: Senhwa Biosciences 




The following authors have nothing to disclose: Kais Zakharia, Katsuyuki Miyabe, 

Catherine D. Moser, Mitesh J. Borad 

397 

Aspirin Use is Associated with Reduced Risk of Cholangiocarcinoma 

Jonggi Choi 1 , Hassan M. Ghoz 1 , Thoetchai Peeraphatdit 1,2 , Esha 

Baichoo 1,3 , Benyam D. Addissie 1 , William S. Harmsen 4 , Terry M. 

Therneau 4 , Janet E. Olson 4,5 , Roongruedee Chaiteerakij 1,6 , Lewis 

R. Roberts 1 ; 1 Division of Gastroenterology and Hepatology, Mayo 

Clinic College of Medicine, and Mayo Clinic Cancer Center, Rochester, 

MN; 2 Department of Internal Medicine, University of Minnesota, 

Minneapolis, MN; 3 Department of Medicine, Mount Sinai 

Roosevelt Hospital, New York, NY; 4 Department of Biomedical 

Statistics and Informatics, Mayo Clinic, Rochester, MN; 5 Division 

of Epidemiology, Department of Health Sciences Research, Mayo 

Clinic, Rochester, MN; 6 Department of Medicine, Faculty of Medicine, 

Chulalongkorn University and King Chulalongkorn Memorial 

Hospital, Thai Red Cross Society, Bangkok, Thailand 

Background & Aims: It is unclear whether aspirin reduces cholangiocarcinoma 

(CCA) risk. Previous studies on risk factors 

for CCA combined all the three CCA subtypes (intrahepatic, 

perihilar, and distal) together or combined perihilar and distal 

CCA as extrahepatic CCA. Whether these factors conferred 

the risk of all 3 subtypes to a similar magnitude is unknown. 

We aimed to determine the associations between aspirin use 

and other risk factors for each CCA subtype individually. Methods: 

We enrolled 2,395 CCA cases (1,169 intrahepatic, 995 

perihilar and 231 distal) seen at Mayo Clinic, Rochester, MN 

between 2000 and 2014. Controls selected from the Mayo 

Clinic Biobank were matched 2:1 with cases by age, sex, 

race, and residential area (n=4,769). Associations between 

aspirin use and other factors and CCA risk were determined 

using logistic regression analysis. Results: There were 591 

(24.7%) CCA cases and 2,129 (44.6%) controls who took 

aspirin. Of aspirin users, 560 (94.8%) of cases and 2008 

(94.3%) of controls were daily users; 384 (65.0%) of cases 

and 1,697 (79.7%) of controls took low-dose aspirin (81-162 

mg/day). Aspirin use had a significant inverse association 

for all CCA subtypes with adjusted odds ratios (AOR) (95% 

confidence interval, 95% CI) of 0.35 (0.29-0.42), 0.34 (0.27- 

0.42), and 0.29 (0.19-0.44), for intrahepatic, perihilar and 

distal CCA, respectively, p


Disclosures: 




The following authors have nothing to disclose: Jonggi Choi, Hassan M. Ghoz, 

Thoetchai Peeraphatdit, Esha Baichoo, Benyam D. Addissie, William S. Harmsen, 

Terry M. Therneau, Janet E. Olson, Roongruedee Chaiteerakij 

398 

The autophagy marker LC3 is significantly associated 

with overall survival in human hepatocellular carcinoma 

underwent resection 

Chih-Wen Lin 1,3 , Jhy-Shrian Huang 1 , Chia-Yen Dai 2,3 , Jee-Fu 

Huang 2,3 , Wan-Long Chuang 2,3 , Ming-Lung Yu 2,3 ; 1 Division of 

Gastroenterology and Hepatology, Department of Medicine, E-DA 

Hospital/ I-SHOU University, Kaohsiung, Taiwan; 2 Hepatobiliary 

Division, Department of Internal Medicine, Kaohsiung Medical 

University Hospital, Kaohsiung Medical University,, Kaohsiung, 

Taiwan; 3 Graduate Institute of Medicine, College of Medicine, 

Kaohsiung Medical University, Kaohsiung, Taiwan 

BACKGROUND: Hepatocellular carcinoma (HCC) is one of 

the most common malignancies, with an increasing incidence 

and is the third leading cause of cancer-related mortality in 

the world. Most HCC patients are diagnosed at an advanced 

stage and have very poor prognosis and low survival. Despite 

the improvements of therapeutic modalities in HCC, the rate of 

5-years tumor-related death has remained as high as at 30-50 

%. Therefore, identification of prognostic factors to develop 

newer targeted therapy is urgently needed to improve HCC 

patient survival. Autophagy is a process through which longlived 

proteins and damaged organelles are conveyed to the 

lysosome for removal by degradation and recycling. Some 

studies have shown that autophagy plays a key role in the 

progression and development of cancer. But, the role of autophagy 

in the prognosis and metastasis of human HCC is not 

well unknown. AIMS: The study aims to explore the expression 

of markers of autophagy genes using immunohistochemistry 

(IHC) in human HCC tissues. We also investigate the autophagy 

markers associated with clinicopathological characteristics 

and prognosis. METHODS: We retrospectively analyzed 

200 patients diagnosed with HCC by histology after surgical 

resection at the Chunghua Christian Hospital, Chunghua, and 

Kaohsiung Medical University Hospital, Kaohsiung, Taiwan, 

from 2009 to 2014. The demographic data, recurrence, and 

survival were collected until December 2014. The expression 

of autophagy-related markers (LC3 and p62) were analyzed 

by IHC staining using HCC tumor tissues and non-tumor tissues. 

RESULTS: Two hundreds HCC patients were collected. 

The average age is 62.8 years old and the rate of male is 

74%. The rate of HBV, HCV, HBV+HCV, and Non-HBVHCV is 

48%, 28%, 4%, and 20%, respectively. Median survival was 

26.3 months (range 3-42 months). The positive rate of LC3 was 

significantly higher in HCC tumor tissues than non-tumor tissues 

(85.5 vs. 50.0%, P < 0.001). In HCC, The overall survival was 

significantly correlated with cirrhosis background, TMN stage, 

BCLC stage, Child-Pugh class, and LC3 tumor part staining in 

univariate Cox regression analysis. Furthermore, the overall 

survival was significantly correlated with cirrhosis background 

(P = 0.01), TMN stage, BCLC stage (P = 0.49), and LC3 tumor 

part staining (P = 0.015) in multivariate Cox regression analysis. 

The strong positive of LC3 staining is significantly associated 

with increasing the 3-year survival rates by Kaplan-Meier 

survival analysis (P=0.046). CONCLUSIONS: Our results show 

that the expression of autophagy marker, LC3, might be a 

strong prognostic factor of overall survival in HCC patients 

underwent liver resection. 

Disclosures: 



The following authors have nothing to disclose: Chih-Wen Lin, Jhy-Shrian Huang, 

Chia-Yen Dai, Jee-Fu Huang, Ming-Lung Yu 

399 

Conversion therapy with hepatic arterial infusion 

chemotherapy and hepatic resection prolongs overall 

survival of patients with advanced hepatocellular carcinoma 

involving vascular invasion 

Hiroaki Nagamatsu 1,2 , Takuji Torimura 2 ; 1 Department of Gastroenterology 

and Hepatology, Yame general hospital, Yame, Japan; 

2 Division of Gastroenterology, Kurume University School of Medicine, 

Kurume, Japan 

[Objectives] Sorafenib is recognized as a standard treatment 

for advanced unresectable hepatocellular carcinoma (HCC) 

with vascular invasion worldwide. However, the therapeutic 

efficacy is marginal. We retrospectively evaluated the therapeutic 

effectiveness of conversion therapy with hepatic arterial 

infusion chemotherapy (HAIC) . [Subjects] From January 2004 

to December 2014, among 270 patients who received HAIC 

with cisplatin (CDDP) and 5-fluorouracil (5-FU) at our institution 

and affiliated Kurume university hospital, 189 patients with 

unresectable HCC involving vascular invasion without extrahepatic 

metastasis were enrolled in the study (mean age, 68.9 

years; Child-Pugh class A/B/C, 114/66/9 cases; portal vein 

tumor thrombosis in 2nd branch/1st branch/trunk, 80/66/43 

cases ). [Methods] All 189 patients received HAIC via the 

reservoir system. After chemolipiodolization, the patients were 

treated with CDDP 50 mg followed by 5-FU 1,250 mg via a 

balloon pump for 5 days (NFP therapy 1) ). For patients who 

achieved partial response (PR) after NFP and in whom conversion 

therapy was feasible, hepatectomy (Hr), trancecatheter 

arterial chemoembolization (TACE), or radiotherapy (RT) was 

performed to achieve cancer-free outcome. Tumor response 

by NFP was evaluated with m-RECIST. Cumulative overall survival 

(OS) after NFP was estimated using Kaplan-Meier survival 

curves, and multivariate analysis was performed to identify 

prognostic factors affecting OS using Cox proportional hazards 

models. For factors with significant difference in the multivariate 

analysis, OS was compared using log-rank test. [Results] 136 

patients (72%) responded to NFP therapy (CR, 17; PR,119). 

41 patients who showed PR received conversion therapy,( 

NFP +Hr, 18 patients; NFP +TACE, 18 patients, NFP +RT, 5 

patients). 58 patients (31%) achieved cancer-free outcome. 

Median OS (MST) after HAIC in all patients was 18 months. 

Significant factors for OS included cancer-free outcome (hazard 

ratio, 0.184; P=0.001) and addition of Hr (hazard ratio, 

0.105; P=0.003). MST in patients who responded was 29 

months, and in patients who achieved cancer-free outcome, it 

was extended to 57 months.Three - and five-year survival rates 

were 78% and 21% in patients treated with NFP only, 87% 

and 71% in NFP+Hr, 47% and 16% in NFP+TACE, and 67% 

and 33% in NFP+RT patients (P=0.028), respectively. [Conclusions] 

NFP therapy demonstrated favorable tumor response in 

patients with advanced hepatocellular carcinoma with vascular 

invasion. Addition of conversion therapy, specifically, Hr, targeting 

cancer-free outcome made possible long-term survival. 

Disclosures: 

The following authors have nothing to disclose: Hiroaki Nagamatsu, Takuji Torimura


400 

Model to predict recurrence after living donor liver 

transplantation for hepatocellular carcinoma beyond 

the Milan criteria 

Yuri Cho 5 , Jeong-Hoon Lee 5 , Eun Ju Cho 5 , Donghyeon Lee 5 , 

Jeong-Ju Yoo 5 , Minjong Lee 5 , Young Youn Cho 5 , Su Jong Yu 5 , 

Nam-Joon Yi 4 , Kwang-Woong Lee 4 , Seong Hoon Kim 1 , Jong Man 

Kim 2 , Jae Won Joh 2 , June Sung Lee 3 , Yoon Jun Kim 5 , Kyung-Suk 

Suh 4 , Jung-Hwan Yoon 5 ; 1 Center for Liver Cancer, National Cancer 

Center, Goyang-Si, Korea (the Republic of); 2 Department of 

Surgery, Samsung Medical Center, Seoul, Korea (the Republic of); 

3 Department of Internal Medicine, Inje University, Ilsan Paik Hospital, 

Goyang-Si, Korea (the Republic of); 4 Department of Surgery, 

Seoul National University College of Medicine, Seoul, Korea (the 

Republic of); 5 Department of Internal Medicine and Liver Research 

Institute, Seoul National University College of Medicine, Seoul, 

Korea (the Republic of) 

Background: Some subgroups of hepatocellular carcinoma 

(HCC) exceeding the Milan criteria (MC) experience substantial 

benefit from living donor liver transplantation (LDLT). This 

study aimed to develop and validate a model to predict tumor 

recurrence after LDLT (MoRAL) for HCC beyond the MC. Methods: 

This multicenter study included a total of 566 consecutive 

patients who underwent LDLT in Korea: the beyond-MC cohort 

(n=205, the derivation [n=92] and validation [n=113] sets) 

and the within-MC cohort (n=361). Results: Using multivariate 

Cox proportional hazard model, we derived the MoRAL score 

(11 x √protein induced by vitamin K absence-II + 2 x √alpha-fetoprotein), 

which provided a good discriminant function on 

time-to-recurrence (c-index=0.88). C-index was maintained similarly 

on both internal and external validations (mean 0.87 and 

0.84, respectively). At cut-off of 314.8 (75th percentile value), 

a low MoRAL score (≤314.8) was associated with significantly 

longer recurrence-free (vs. >314.8, hazard ratio [HR]=5.29, 

p


right upper quadrant pain (24%), and hepatomegaly (24%). 

Routine screening detected carcinoma in 9/25 patients; 8/9 

of those screened cases were caught at an early stage with a 

single tumor. Prior to diagnosis with HCC, the median interval 

since prior imaging was 12 (1-42) months. Patients without 

prior or recent (within 12 months) liver imaging were more 

likely to present with multiple liver masses or metastatic disease 

(91%). Moreover, 88% of patients deceased before 1 year 

after diagnosis had no prior or recent screening. Conclusion: 

Patients with Fontan circulation are at increased risk to develop 

late hepatocellular cancer and should be screened according 

to guidelines. Early diagnosis of hepatocellular carcinoma may 

allow for improved treatment strategies and prolonged survival. 

Presenting Stages of Hepatocellular Carcinoma 

for Epi-TACE group and 869 days for Cis-TACE group (HR: 

1.18, 95% CI:0.57-2.44, p = 0.658). In subgroup analysis for 

the primary treatment, response rate was significantly better in 

Epi-TACE group than in Cis-TACE group (64.3% versus 8.3%, 

respectively, p = 0.002), though they were comparable in 

subgroup analysis for the non-primary treatment (16.7% versus 

26.7%, respectively, p = 0.531). Conclusions This prospective 

randomized controlled study demonstrated that Epi-TACE might 

be more effective than Cis-TACE. Epirubicin might be used 

for the primary TACE, and cisplatin might be alternatives as 

non-primary TACE. 

Disclosures: 

Etsuro Hatano - Speaking and Teaching: Bayer 

The following authors have nothing to disclose: Yosuke Kasai, Kohta Iguchi, 

Rinpei Imamine, Takahiro Nishio, Masayuki Okuno, Satoru Seo, Kojiro Taura, 

Kentaro Yasuchika, Toshiya Shibata, Shinji Uemoto 

Disclosures: 

Daniel Ganger - Advisory Committees or Review Panels: Bristol Myers, Abbvie; 

Grant/Research Support: Merck, Ocera; Speaking and Teaching: Gilead 

The following authors have nothing to disclose: Laura J. Dickmeyer, Barbara J. 

Deal, Andrew Defreitas 

402 

A randomized controlled study of effectiveness between 

transcatheter arterial chemoembolization with epirubicin 

versus cisplatin for multiple hepatocellular carcinoma 

Yosuke Kasai 1 , Etsuro Hatano 1 , Kohta Iguchi 1 , Rinpei Imamine 2 , 

Takahiro Nishio 1 , Masayuki Okuno 1 , Satoru Seo 1 , Kojiro Taura 1 , 

Kentaro Yasuchika 1 , Toshiya Shibata 2 , Shinji Uemoto 1 ; 1 Department 

of Surgery, Graduate School of Medicine, Kyoto University, 

Kyoto, Japan; 2 Department of Diagnostic Imaging and Nuclear 

Medicine, Graduate School of Medicine, Kyoto University, Kyoto, 

Japan 

Background and Aim In transcatheter arterial chemoembolization 

(TACE) for hepatocellular carcinoma (HCC), it has not been 

clarified which chemotherapy agent is an effective combined 

drug with lipiodol. We previously showed that lipiodol-TACE 

combined with cisplatin (Cis-TACE) provided a higher response 

rate than lipiodol-TACE combined with epirubicin (Epi-TACE) 

for multiple HCC in a retrospective cohort study (Hepatol Res. 

2011). We conducted a randomized controlled study comparing 

the effectiveness of Epi-TACE and Cis-TACE. Method 

Sixty patients with multiple HCC which were not indicated for 

curative liver resection or percutaneous ablation therapy were 

randomly assigned to Epi-TACE or Cis-TACE group, stratified 

by the primary or non-primary treatment and the presence or 

absence of the history of liver resection. Finally, 26 patients for 

Epi-TACE group and 27 patients for Cis-TACE group met the 

eligibility criteria and were analyzed. The primary endpoint 

was response rate, and the secondary endpoints were progression 

free survival (PFS) and overall survival (OS). Results 

Patient characteristics were almost comparable between the 

two groups, except for larger tumor size in Epi-TACE group. 

The response rates were 42.3% for Epi-TACE group and 

18.5% for Cis-TACE group (p = 0.057). When adjusted by 

tumor size, the odds ratio for Epi-TACE group against Cis-TACE 

group was 3.47 [95% confidence interval (CI): 0.98-13.6, p = 

0.054]. The median PFS times were 224.5 days for Epi-TACE 

group and 165 days for Cis-TACE group [hazard ratio (HR) for 

Epi-TACE group against Cis-TACE group: 0.83, 95% CI: 0.47- 

1.45, p = 0.503], and the median OS times were 1128 days 

403 

Overexpression of periostin is related to poor prognosis 

of hepatocellular carcinoma 

Se Young Jang 1 , Soo Young Park 1 , Won Young Tak 1 , Young Oh 

Kweon 1 , Keun Hur 2 , Gyeonghwa Kim 2 , Yong-Hun Choi 2 , Jung Gil 

Park 3 , Yu Rim Lee 1 , Sun Kyung Jang 1 , Su Hyun Lee 1 ; 1 Kyungpook 

National University Hospital, Daegu, Korea (the Republic of); 2 Biochemistry 

and Cell Biology, Kyungpook National University School 

of Medicine, Daegu, Korea (the Republic of); 3 Gastroenterology 

and Hepatology, CHA University, Gumi CHA medical center, 

Gumi, Korea (the Republic of) 

Background: Periostin is an extracellular matrix protein and 

known to be related to the metastatic potential and prognosis 

of human cancers in recent studies. However, few studies 

showed the expression level of periostin and its association 

with prognosis of hepatocellular carcinoma (HCC). We analyzed 

periostin overexpression and its implication for prognosis 

of HCC. Methods: A total of 149 patients who underwent 

surgical resection in Kyungpook National University Hospital 

between 2006 and 2010 were selected and retrospectively 

reviewed. All tissue specimens were formalin-fixed and paraffin-embedded 

for immunohistochemistry staining. A tissue 

microarray (TMA) containing tissue from HCC (n=149), adjacent 

non-tumor tissues of HCC. Results: Expression of periostin 

was associated with microvascular invasion (OR=3.383; 

p=0.001), multiple tumors (OR=2.631; p=0.020), and higher 

modified UICC stage (OR=2.945, p=0.004). The patients with 

positive periostin expression have significantly shorter overall 

survival (p=0.021) and recurrence-free survival (p=0.032). 

Cumulative overall survival was significantly different between 

periostin negative and positive groups when microvascular 

invasion was absent (p=0.018). Conclusion: We found that 

high periostin expression on hepatocellular carcinoma tissues is 

correlated with poor prognosis. Especially when microvascular 

invasion is absent, periostin can be a better prognostic marker 

for overall survival.


Cumulative overall survival rates of periostin positive and negative 

gruops according to the existence of microvascular invasion. 

Disclosures: 

Won Young Tak - Advisory Committees or Review Panels: Gilead Korea; Grant/ 

Research Support: SAMIL Pharma; Speaking and Teaching: Bayer Korea 

The following authors have nothing to disclose: Se Young Jang, Soo Young Park, 

Young Oh Kweon, Keun Hur, Gyeonghwa Kim, Yong-Hun Choi, Jung Gil Park, 

Yu Rim Lee, Sun Kyung Jang, Su Hyun Lee 

404 

Impact of screening on Hepatocellular Carcinoma survival 

in patients with Chronic Hepatitis B - A regional 

population based study 

Rasham Mittal, Bechien U. Wu; Kaiser Permanente Los Angeles 

Medical Center, Los Angeles, CA 

Background: The impact of screening for hepatocellular carcinoma 

(HCC) in patients with chronic hepatitis B (CHB) remains 

controversial. Aim: To determine if screening is associated with 

improved survival among CHB patients with HCC in a diverse 

regional U.S.population. Methods: Retrospective cohort study 

(2006-2013) from an integrated health care system in Southern 

California. All CHB (age ≥18 years) patients identified 

using ICD codes and confirmed by serology. HCC patients 

were identified using ICD code and cross referenced through 

prospective internal cancer registry. Patients were considered 

to have undergone screening if they had USG, CT scan or MRI 

abdomen with contrast imaging at 6-18 months prior to HCC 

diagnosis. Patients were excluded if they did not have active 

Kaiser Membership for at least 18 months prior to HCC diagnosis. 

Survival curves estimated using Kaplan-Meyer analysis. 

Cox proportional hazard regression analysis was performedt o 

adjust for baseline characteristics. Results: From a total of 9811 

CHB patients, we identified 185 HCC cases [median age at 

HCC diagnosis 61 years, IQR 53, 68 years; 147 (79.4%) 

male; 136 (73.5%) Asians; 123 (66.5%) liver cirrhosis]. Out 

of 185, 110 patients had screening prior to HCC while 75 presented 

with HCC without prior screening. The median observed 

survival was higher in screening versus unscreened group (5.9 

years versus 3.2 years; P=0.036). In regression analysis, HCC 

screening (HR 0.60; 95%CI, 0.37-0.96; P=0.03) were associated 

with improved survival after adjusting for baseline characteristics 

such as age (HR 0.97; 95%CI, 0.95-0.99; P=0.259), 

female gender (HR 0.40; 95%CI, 0.20-0.85; P=0.012) Charlson 

co-morbidity score (HR 1.14; 95%CI, 1.04-1.24; P=0.003) 

and liver cirrhosis (HR 1.14; 95%CI, 0.66-1.98; P=0.623). 

Conclusion: HCC surveillance was associated with improved 

survival in CHB patients in a large regional U.S. population. 

Disclosures: 

The following authors have nothing to disclose: Rasham Mittal, Bechien U. Wu 

405 

Influence of hepatitis B virus DNA elevation on recurrence 

of hepatocellular carcinoma after surgical resection 

and the preventive role of antiviral therapy 

Yang J. Yoo, Ji Hoon Kim, Seong Hee Kang, Young-Sun Lee, Tae 

Suk Kim, Sang Jun Suh, Young Kul Jung, Yeon Seok Seo, Hyung 

Joon Yim, Jong Eun Yeon, Kwan Soo Byun; Department of Internal 

Medicine, Korea University College of Medicine, Seoul, Korea (the 

Republic of) 

Aims Surgical resection is the treatment of choice for early 

stage hepatocellular carcinoma (HCC). Previous studies 

revealed that reactivation of hepatitis B virus is associated with 

the recurrence of hepatitis B virus (HBV) related HCC after surgical 

resection. We aimed to investigate the influence of HBV 

DNA elevation on HCC recurrence and the preventive role of 

antiviral therapy. Methods One hundred eleven patients who 

had BCLC stage 0 or A and received surgical resection as primary 

therapy were enrolled. HBV DNA elevation was defined 

as reactivation (increase >1log 10 

IU/ml or re-emergence of 

HBV DNA) in patients without preoperative antiviral therapy or 

virologic breakthrough in patients with preoperative antiviral 

therapy (n=62). Results Mean age was 54.1±9.4 years and 

male consisted 76.6% (n=85). All patients belonged in Child 

class A. BCLC 0 consisted 29.7% (n=33) and 30.6% (n=34) 

had Hepatitis B envelope antigen positivity and 42.3% (n= 

47) had HBV DNA level ≥ 2000 IU/mL at the time of surgery. 

Overall 1 year, 3 year, 5 year recurrence was 17.3%, 36.4%, 

40%. In multivariate analysis for risk factors of recurrence, 

multiple tumor, HBV DNA elevation, ALT ≥ 30 IU/L, were independently 

associated with HCC recurrence. In subgroup analysis, 

patients with preoperative antiviral therapy showed similar 

HCC recurrence rate with patients who start antiviral therapy 

after resection and significantly higher recurrence rate than 

patients with no antivirals during follow up. In multivariate analysis 

for risk factors of HBV DNA elevation, Age < 50 years, 

no preoperative antiviral therapy, HBV DNA ≥ 2000 remained 

as risk factors. To rule out the antiviral effect, we investigated 

risk factors for HCC recurrence in each group with or without 

preoperative antiviral therapy. In patients with preoperative 

antiviral therapy, multiple tumor was the only risk factor for 

HCC recurrence. In patients without preoperative antiviral therapy, 

male sex and HBV DNA elevation were independent risk 

factors for HCC recurrence. Risk factors for HBV DNA elevation 

were further analyzed in patients without preoperative antiviral


therapy. Age < 50 years and HBV DNA ≥ 2000 IU/mL were 

independent risk factors for HBV DNA elevation. Conclusion 

HBV DNA elevation after resection increases the risk of HCC 

recurrence irrespective of preoperative antiviral therapy. However, 

HBV DNA elevation is an independent risk factor for 

recurrence in patients without preoperative antiviral therapy. 

Therefore, perioperative antiviral therapy should be considered 

to prevent HBV DNA elevation and recurrence, especially in 

patients with Age < 50 years and/or HBV DNA ≥ 2000 IU/ 

mL. 

Disclosures: 


The following authors have nothing to disclose: Yang J. Yoo, Ji Hoon Kim, Seong 

Hee Kang, Young-Sun Lee, Tae Suk Kim, Sang Jun Suh, Young Kul Jung, Yeon 

Seok Seo, Hyung Joon Yim, Jong Eun Yeon 

survival (P = 0.024). Conclusions: Gene alterations in TERT 

promoter, TP53, CTNNB1, and HBV integration were closely 

associated with HCC development, and mutations in TERT promoter 

are related to poor prognosis. These results are useful for 

understanding the underlying mechanism of hepatocarcinogenesis, 

diagnosis, and predicting outcomes of patients with HCC. 

Disclosures: 

Sei Kakinuma - Grant/Research Support: The Japanese Society of Gastroenterology, 

MSD Co., Ltd. 

Mamoru Watanabe - Grant/Research Support: MSD Co., Ltd. 

The following authors have nothing to disclose: Fukiko Kawai-Kitahata, Yasuhiro 

Asahina, Shinji Tanaka, Miyako Murakawa, Sayuri Nitta, Takako Watanabe, 

Satoshi Otani, Fumio Goto, Hiroko Nagata, Shun Kaneko, Seishin Azuma, 

Yasuhiro Itsui, Mina Nakagawa, Minoru Tanabe, Shinya Maekawa, Nobuyuki 

Enomoto 

406 

Comprehensive analyses of mutations and hepatitis B 

virus integration in hepatocellular carcinoma with clinicopathological 

features 

Fukiko Kawai-Kitahata 1 , Yasuhiro Asahina 1 , Shinji Tanaka 1 , Sei 

Kakinuma 1 , Miyako Murakawa 1 , Sayuri Nitta 1 , Takako Watanabe 

1 , Satoshi Otani 1 , Fumio Goto 1 , Hiroko Nagata 1 , Shun 

Kaneko 1 , Seishin Azuma 1 , Yasuhiro Itsui 1 , Mina Nakagawa 1 , 

Minoru Tanabe 1 , Shinya Maekawa 2 , Nobuyuki Enomoto 2 , 

Mamoru Watanabe 1 ; 1 Tokyo Medical and Dental University, 

Tokyo, Japan; 2 University of Yamanashi, Yamanashi, Japan 

Background & Aims: Genetic alterations in specific genes 

are critical events in carcinogenesis and hepatocellular carcinoma 

(HCC) progression. However, the genetic alterations 

responsible for HCC development, progression, and survival 

are unclear. Methods: The extracted DNA from 104 patients 

was amplified by multiplexed PCR targeting 54 cancer-related 

genes containing 2820 mutational hotspots. The mutations 

were analyzed using a semiconductor-based DNA sequencer. 

Nucleotide variants that were detected in tumors and absent 

in corresponding non-tumor tissue were determined as somatic 

mutations. We also investigated TERT promoter mutation at 

2 hot spots through direct sequencing. HBV oligonucleotide 

probes were designed primarily within the conserved regions 

of different HBV strains including genotypes Aa, Ae, Ba, Bj, 

and C. After incubation with custom capture oligos, hybridized 

fragments were sequenced on a MiSeq instrument generating 

300 bp paired-end reads. Results: We found recurrent 

mutations in several genes such as TERT (65%), TP53 (38%), 

CTNNB1 (30%), AXIN1 (2%), PTEN (2%), and CDKN2A 

(2%). TERT promoter mutations were associated with older 

age (P = 0.005), presence of hepatitis C virus (HCV) infection 

(P = 0.003), and absence of hepatitis B virus (HBV) infection 

(P < 0.0001). In patients with hepatitis B surface antigen 

(HBsAg) positive, a total of 78 HBV integration breakpoints 

were detected in 25/27 (92.6%) patients. Most (89%) of HBV 

integrant was the HBx region. HBV integration was frequently 

found in particular genes including TERT (n = 9), KMT2B (MLL4; 

n = 2), and MYO7A (n = 2). Seven (77.8%) of TERT locus 

breakpoints were paired with HBx gene, all at the 3’-end of 

HBx gene. HBV integration into TERT locus was mutually exclusive 

to TERT promoter mutations. The HBV integration number 

was significantly associated with HCC at younger age (Spearman’s 

rank order correlation, r s 

= −0.5342, P < 0.01). TP53 

mutations were associated with HBV infection (P = 0.0001) 

and absence of HCV infection (P = 0.002). CTNNB1 mutations 

were associated with absence of HBV infection (P = 0.010). 

Moreover, TERT promoter mutation was significantly associated 

with shorter disease-free survival (P = 0.005) and poor overall 

407 

Treatment not needed for hypovascular tumors associated 

with hepatocellular carcinoma 

Yutaka Midorikawa, Tadatoshi Takayama; Nihon Univ., Tokyo, 

Japan 

Aim: Hypovascular tumors associated with hepatocellular 

carcinoma (HCC) can be diagnosed, but it remains unknown 

whether such lesions should be treated immediately after diagnosis. 

This study aimed to clarify whether treating hypovascular 

liver nodules contributes to the prolongation of patient survival, 

and sought to directly calculate a lead time from the diagnosis 

of hypovascular liver nodules to their transformation into 

hypervascular HCC. Methods: One hundred and four patients 

with hypovascular liver nodules smaller than 3 cm underwent 

liver resection immediately after diagnosis (Group 1), while 

80 patients with hypovascular liver nodules were treated after 

appearance of vascularization or other hypervascular HCC 

lesions on imaging studies (Group 2). To avoid lead-time bias 

for tumor vascularization, the survival rates of patients after 

diagnosis of hypovascular liver nodules as well as the survival 

rates after liver resection in the two groups were compared. 

Results: After a median follow-up period of 3.3 years (range, 

0.6–11.2), the median overall survival rates “after liver resection” 

were 9.7 years (95% confidence interval [CI], 6.7–11.9) 

and 8.5 years (4.8–12.1; P = 0.017), respectively, and recurrence-free 

survivals were 3.8 years (95% CI, 2.7–6.3) and 1.9 

years (1.5–2.9; P


Disclosures: 

The following authors have nothing to disclose: Yutaka Midorikawa, Tadatoshi 

Takayama 

408 

Elevated Serum Wisteria floribunda agglutinin-positive 

human Mac-2 binding protein predict all-cause mortality 

in hepatitis C patients with hepatocellular carcinoma 

Shigemune Bekki, Kazumi Yamasaki, Yuuki Sonoda, Yuki Kugiyama, 

Satoru Hashimoto, Akira Saeki, Shinya Nagaoka, Seigo 

Abiru, Atsumasa Komori, Hiroshi Yatsuhashi; National Organization 

Hospital Nagasaki Medical Center, Omura, Japan 

Background & Aims: Serum glycosylated Wisteria floribunda 

agglutinin-positive Mac-2 binding protein (WFA(+)-M2BP) 

has been recently reported as a noninvasive and useful surrogate 

marker for the degree of liver fibrosis and for the risk of 

hepatocellular carcinoma (HCC). We investigated the utility 

of WFA(+)-M2BP to predict all-cause mortality in the patients 

with hepatitis C virus (HCV)-related hepatocellular carcinoma 

(HCC). Patients & Methods: A total of 466 HCV-related HCC 

patients who had been admitted to our hospital from 1999 

to 2013 were enrolled. We evaluated association between 

mortality and clinicolaboratory variables, included age, sex, 

asparate and alanine aminotransferase levels (AST and ALT), 

bilirubin, albumin, prothrombin activity, platelet count, Child- 

Pugh grade, alpha-fetoprotein (AFP), des-γ-carboxy prothrombin 

(DCP), TNM stage, and WFA(+)-M2BP. The average 

observation period was 3.7 years. The association between 

WFA(+)-M2BP and mortality was evaluated using the Cox proportional 

hazards regression analysis. Results: The baseline 

characteristics of the 466 patients were 295 (63.2%) male with 

a median age of 70.0 years. The median value of WFA(+)- 

M2BP was 4.6 (range, 0.2-18.36) cutoff index (COI). Serum 

WFA(+)-M2BP levels significantly correlated with bilirubin 

(r=0.26, p


410 

Significant Suppression of Local Tumor Recurrence in 

Treatment for Hepatocellular Carcinoma with Peritumoral 

Vessels by Radiofrequency Ablation with Percutaneous 

Ethanol Injection: Propensity Score Matching 

Analysis 

Kazuhide Takata 1 , Akira Anan 1 , Takashi Tanaka 1 , Keiji 

Yokoyama 1 , Daisuke Morihara 1 , Yasuaki Takeyama 1 , Makoto 

Irie 1 , Satoshi Shakado 1 , Kaoru Iwata 2 , Tetsuro Sohda 1 , Shotaro 

Sakisaka 1 ; 1 Gastroenterology and Medicine, Fukuoka University 

Faculty of Medicine, Fukuoka, Japan; 2 Internal medicine, 

Meotoiwa Hospital, Fukuoka, Japan 

Background and Aims: Radiofrequency ablation (RFA) is a 

major modality for the treatment of early stage hepatocellular 

carcinoma (HCC). However, serious problems of RFA remain 

in local recurrence of HCC with peritumoral vessels, probably 

due to heat-sink effect. Therefore, we have undergone percutaneous 

ethanol injection (PEI) to the peritumoral vessels side of 

the tumor to complete tumor necrosis by ethanol, with RFA. The 

aim of the study was to evaluate the effect PEI combination RFA 

for HCC with peritumoral vessels. Methods: From Jun 2001 to 

Mar 2015, 371 patients, who diagnosed newly HCC (total 

525 nodules), 3 cm or less and within 3 nodules, were treated 

RFA as an initial treatment in our hospital. Among 190 nodules 

with peritumoral vessels located less than 5 mm from the tumor, 

patients in group of RFA monotherapy (61 nodules) or group 

of PEI combination (61 nodules) were 1 : 1 matched according 

to their propensity scores. Ethanol was injected inside the 

tumor close to the peritumoral vessels in the combination therapy 

group. Cumulative incidences of HCC local recurrence 

were analyzed. Results: The overall median follow-up period 

was 42 months (range, 3-134). The cumulative HCC local 

recurrence rates were 7.2%, 18.7%, and 24.1% at 1, 3, 5 

years, respectively, in the PEI combination group, and 15.1%, 

34.9%, 46.9% in the RFA monotherapy group; showing significantly 

lower recurrence rates in PEI combination group (P < 

0.05). In Cox regression analysis, presence of PEI combination 

was significantly associated with the risk reduction of HCC 

local recurrence (hazard ratio, 0.44; 95% CI: 0.21, 0.88; P < 

0.05). Conclusions: RFA combined with PEI reduced the risk of 

a local recurrence rate by half, compared with RFA alone, in 

early stage HCC with peritumoral vessels. Further prospective 

studies based on cumulative local recurrence rate may be helpful 

in improving the quality of RFA for HCC. 

Disclosures: 

The following authors have nothing to disclose: Kazuhide Takata, Akira Anan, 

Takashi Tanaka, Keiji Yokoyama, Daisuke Morihara, Yasuaki Takeyama, Makoto 

Irie, Satoshi Shakado, Kaoru Iwata, Tetsuro Sohda, Shotaro Sakisaka 

411 

Comparison of balloon-occluded Transarterial chemoembolization 

(TACE) with conventional TACE for treatment 


Takuro Niinomi, Masatoshi Ishigami, Yoji Ishizu, Teiji Kuzuya, 

Takashi Honda, Kazuhiko Hayashi, Yoshiki Hirooka, Hidemi Goto; 

Gastroenterology and Hepatology, Nagoya Univercity Graduate 

School of Medicine, Nagoya, Japan 

AIM: Transarterial chemoembolization (TACE) has been widely 

performed as a treatment for hepatocellular carcinoma (HCC) 

in patients who don’t have the indication for curative treatment. 

Selecting the anti-cancer agents, treatment methods and equipment 

for the best therapeutic effect is becoming more important 

for performing better quality of TACE. In recent years, balloon-occluded 

TACE (B-TACE) by using microballoon catheters 

with small diameters has been widely applied, primarily in 

Japan. The purpose of this study was to compare the early 

therapeutic efficacy and lipiodol deposition of B-TACE with 

those of conventional TACE (C-TACE). Patients and Methods: 

From April 2013 to November 2014, 200 consecutive patients 

with unresectable HCC were treated with TACE at our institution. 

The patients were divided into two groups based on the 

usage of microballoon catheter. Treatment efficacy and lipiodol 

deposition were retrospectively compared between two 

groups. Of these patients, 20 patients with 51 nodules were 

treated with B-TACE and 39 patients with 99 nodules were 

treated with conventional TACE (C-TACE). Mean lesion density 

and lipiodol emulsion concentration ratio of HCC to embolized 

liver parenchyma (LECHL ratio) in Hounsfield units (HU) were 

evaluated on non-enhanced CT imaging immediately after 

TACE. The treatment effect of TACE was evaluated by dynamic 

CT 1 to 3 months after TACE, and was evaluated based on 

the Response Evaluation Criteria in Cancer Study Group of 

Japan (RECICL2009). Results: The median lesion density on CT 

immediately after TACE was higher in B-TACE group than in 

C-TACE group (B-TACE 533 HU; range 121-1117 HU versus 

C-TACE 422 HU; range 136-1322 HU, P=0.057, Mann-Whitney 

U test). LECHL ratio on CT immediately after TACE was 

remarkably higher in B-TACE group than in C-TACE group 

(B-TACE 3.69; range 1.29-9.7 versus C-TACE 2.82; range 

0.96-12.83, P=0.017, Mann-Whitney U test). The treatment 

effect on the target nodule classified as TE4, TE3, TE2 and 

TE1 were 68%, 16%, 16% and 0% in the B-TACE group, and 

58%, 9%, 28% and 5% in the C-TACE group respectively. 

Response rate (TE4+TE3) was significantly higher in B-TACE 

group than in C-TACE group (84% versus 67%, P=0.0216, 

Fisher’s exact test). Conclusion: Mean lesion density was higher 

in B-TACE group than in C-TACE group. Notably, LECHL ratio 

which imply selective deposition of lipiodol, were significantly 

higher in B-TACE group than in C-TACE group. These results 

suggested that compared with C-TACE, B-TACE significantly 

improved lipiodol deposition on HCC nodule during TACE 

procedure, and could obtain better treatment efficacy. 

Disclosures: 

Hidemi Goto - Grant/Research Support: MSD, Roche, Bayer, Bristol-Myers, Eisai, 

Ajinomoto, Otsuka, Astra, Tanabe, Takeda 

The following authors have nothing to disclose: Takuro Niinomi, Masatoshi 

Ishigami, Yoji Ishizu, Teiji Kuzuya, Takashi Honda, Kazuhiko Hayashi, Yoshiki 

Hirooka 

412 

Hepatic Decompensation After Transarterial Chemoembolization 

For Hepatocellular Carcinoma 

Dempsey Hughes, Fredric D. Gordon, Keith E. Stuart, FW Nugent, 

Sebastian Flake, Mary Ann Simpson, Amir A. Qamar; Transplantation, 

Lahey Hospital and Medical Center, Burlington, MA 

Background:Hepatic decompensation(HD)is a concern in 

patients with cirrhosis who undergo transarterial chemoembolization(TACE)for 

hepatocellular carcinoma.Identifying predictors 

associated with treatment-related HD would allow better 

risk assessment risk associated with TACE.The aim of this study 

is to assess HD after TACE and to identify any pre-treatment 

factors related to HD. Methods:A retrospective cohort study 

of 184 patients with cirrhosis and HCC who underwent TACE 

at our institution from 2009-14 was analyzed.TACE-related 

HD was defined as:1)new onset variceal hemorrhage(VH), 

ascites or hepatic encephalopathy(HE), 2)treatment change 

for known history of VH,ascites (increased diuretic or paracentesis 

requirement), HE(increase or addition of lactulose or 

rifaximin), or spontaneous bacterial peritonitis within 90 days 

of first TACE.TACE was performed using standard protocol.


Univariate analysis compared baseline factors.Baseline factors 

independently associated with HD after TACE were identified 

with Cox regression. Results:51 patients(28%)experienced an 

HD event within 90 days of TACE.Median time to decompensation 

event after first TACE was 52 days.On univariate analysis, 

alkaline phosphatase(ALP), total bilirubin(TB), albumin, 

INR, MELD, being within Milan or UCSF criteria, history of 

HD event within 1 year of TACE significantly differed between 

decompensated and compensated groups(TABLE).Cox regression 

model identified a decompensation event within 1 year 

prior to TACE(HR 2.19,p=0.02), ALP(HR 1.003,p=0.007)and 

being outside UCSF criteria(HR 0.49,p=0.02)as independently 

associated with an HD event after TACE.TB, albumin, INR and 

MELD were not. Conclusion:For patients with cirrhosis being 

considered for TACE, absence of decompensating event within 

1 year of TACE, HCC within UCSF criteria and low ALP are 

associated with low risk of TACE-related HD. 

Univariate Analysis 

Disclosures: 

The following authors have nothing to disclose: Dempsey Hughes, Fredric D. 

Gordon, Keith E. Stuart, FW Nugent, Sebastian Flake, Mary Ann Simpson, Amir 

A. Qamar 

between pathological and radiological findings regarding 

the number of nodules was found in 103/184(56%): 78 

were underestimated and 25 overestimated by radiology. 

10/40(25%) patients who were within Milan criteria, resulted 

outside according to histology due to discrepancy in number 

of nodules. Considering the diameter of the biggest nodule on 

explant, discordance was observed in 158/181(87%): 93 

were underestimated and 65 overestimated. 20 of 40(50%) 

within Milan criteria on imaging, exceeded these due to 

discrepancy in biggest nodule diameter. HCC recurrence 

occurred in 29/184(16%) patients in a median of 37m (4-157) 

post-LT. In Cox regression, pre-LT factors associated with HCC 

recurrence were aFP>100 IU/ml (p3cm 

(p3cm but different 

aFP levels (22% in aFP100, p=0.003). 

The same pre-LT factors were also significantly associated with 

HCC-related mortality post-LT. Conclusion: In HCC transplanted 

patients there was a marked discrepancy between radiological 

findings and pathological description of the explant. The best 

predictors of HCC recurrence were pre-LT aFP and diameter 

of the biggest nodule. Combined together could allow us to 

include in the current transplant criteria patients with biggest 

nodule diameter>3cm if aFP


on explant (>3 tumors, largest tumor >5cm, vascular invasion, 

metastases, or poor differentiation). NASH patients were less 

likely to undergo locoregional therapy (LRT) pre-LT (86 vs 92%; 

p


was 53 +/-0.6 days. By multivariate analysis, factors related 

to all causes of death were patients’ age >60 years (OR=1.2, 

95%CI; 1.1-1.3), length of hospital stay of >7 days (OR=1.1, 

95%CI; 1.02-1.2), male (OR=1.3, 95%CI; 1.2-1.4), living 

in Northern part of Thailand (OR=1.5, 95%CI; 1.3-1.8) and 

presence of complications during admission (OR=1.3, 95%CI; 

1.1-1.5. CONCLUSIONS: The disease burden of patients with 

cholangiocarcinoma in Thailand is significant with 0.65% of 

in-patients with GI. diseases and high mortality rate of 14%. 

Disclosures: 

The following authors have nothing to disclose: Sombat Treeprasertsuk, Kittiyod 

Poovorawan, Ngamphol Soonthornworasiri, Roongruedee Chaiteerakij, Pisaln 

Mairiang, Kookwan Sawadpanich, Varocha Mahachai, Kamthorn Phaosawasdi 

417 

Short-term preoperative treatment with sorafenib for 

patients with resectable hepatocellular carcinoma (HCC): 

results of the BIOSHARE neoadjuvant phase II study 

from GERCOR IRC 

Mohamed Bouattour 1 , Laetitia Fartoux 2 , Olivier Rosmorduc 2 , Eric 

Vibert 3 , Charlotte E. Costentin 4 , Olivier Soubrane 5 , Olivier Scatton 

6 , Maxime Ronot 7 , Laurent Castera 1 , Muriel Garnier 8 , Armand 

De Gramont 8 , Jacques Belghiti 5 , Valerie Paradis 9 , Annemilaï 

Tijeras-Raballand 10 , Alexandra Hadengue 11 , David Brusquant 11 , 

Benoist Chibaudel 11 , Eric Raymond 8 , Sandrine Faivre 8 ; 1 Hepatology, 

Beaujon University Hospital, Clichy, France; 2 Hepatology, 

Saint-Antoine University Hospital, Paris, France; 3 Hepato-Biliary 

Surgery, Paul-Brousse University Hospital, Villejuif,, France; 4 Hepatology, 

Henri-Mondor University Hospital, Creteil, France; 5 Hepato-Biliary 

Surgery, Beaujon University Hospital, Clichy, France; 

6 Hepato-Biliary Suregry, Saint-Antoine University Hospital, Paris, 

France; 7 Radiology, Beaujon University Hospital, Clichy, France; 

8 Medical Oncology, Centre Hospitalo-Univesitaire Vaudois 

(CHUV), Lauzanne, Switzerland; 9 Pathology, Beaujon University 

Hospital, Clichy, France; 10 AREC FILIA RESEARCH,, Paris, France; 

11 GERCOR, Paris, France 

Background. Neoadjuvant therapy offers the opportunity of 

investigating new agents in early stage and allows direct 

access to tumor biology. This open-label, multicenter, phase 

II study aimed at investigating the activity of sorafenib including 

radiological, pathological and molecular changes in tumor 

from patients with operable HCC. Patients and methods. A preoperative 

treatment consisting of 400 mg bid orally sorafenib 

was administered for 4 consecutive weeks followed by surgery 

1 week after sorafenib discontinuation. Primary endpoints 

were antitumor activity and histological changes between 

paired biopsies and plasma biomarkers, from baseline and 

post sorafenib treatment. Secondary endpoints were safety 

profile, R0 surgery and post-surgical complications. Residual 

disease characteristics were analyzed on surgical specimens. 

Results. Among 30 patients enrolled, 28 were evaluable for 

safety and 25 patients (21 men; median age: 61.5 years) 

were evaluable for the primary endpoint. All evaluable patients 

were Child Pugh A, 14 (56%) with chronic liver disease, 9 

(36%) with cirrhosis and 2 patient (8%) with no underlying 

liver disease. The baseline median tumor size was 37 mm 

(17-220 mm) and 21 patients (84%) had a single lesion. The 

median duration and dosing of sorafenib for evaluable patients 

were 28 days (21-35d) and 793 mg/day (477 – 843 mg/ 

day) respectively. Overall, the safety profile of preoperative 

sorafenib was good. According to RECIST criteria, all patients 

showed stable disease after 4 weeks of sorafenib. Among 14 

patients evaluated according to mRECIST and Choi criteria, 

objective responses were observed for 4 (29%) and 7 (50%) 

patients respectively. All evaluable patients went on liver resection; 

median hospitalization duration was 8 days (4-59d) and 

no unexpected postoperative complication was observed. R0 

tumor resection was achieved in 22 patients (88%). Surgical 

specimen showed macrovascular and microvascular invasion 

as well as satellite nodules in 12%, 48%, and 36% respectively. 

Intratumor necrosis was observed in 13 patients of 20 

evaluable patients. Biomarkers from pre- and post-treatment 

tissue (Vimentin, CK19, E-Cadherin, N-Cadherin, CK7, MIB1, 

CD31, VEGF, CD133, CA9, p-S6, c-MET, CXCR4) and plasma 

(VEGF-C, Ang2, PlGF, c-KIT, SDF-1, HGF, TGFb1) are currently 

assessed and will be presented during the meeting. Conclusion. 

Preoperative BIOSHARE trial showed appropriate safety 

regarding both tolerance to sorafenib and surgical procedures. 

Short-term neoadjuvant treatment with sorafenib showed promising 

efficacy in terms of tumor response (mRECIST and Choi) 

and will allow detailed evaluation of molecular changes in 

patient tumor tissues. 

Disclosures: 

Mohamed Bouattour - Advisory Committees or Review Panels: Bayer Schering 

Pharma; Speaking and Teaching: Bayer Schering Pharma 

Olivier Rosmorduc - Advisory Committees or Review Panels: Syrtex, IPSEN; 

Speaking and Teaching: Bayer 

Laurent Castera - Speaking and Teaching: Gilead, BMS, Janssen, Echosens, 

Abbvie, Biopredictive 

The following authors have nothing to disclose: Laetitia Fartoux, Eric Vibert, 

Charlotte E. Costentin, Olivier Soubrane, Olivier Scatton, Maxime Ronot, Muriel 

Garnier, Armand De Gramont, Jacques Belghiti, Valerie Paradis, Annemilaï 

Tijeras-Raballand, Alexandra Hadengue, David Brusquant, Benoist Chibaudel, 

Eric Raymond, Sandrine Faivre 

418 

Analytic Morphomics Predicts Survival in Patients with 

Hepatocellular Carcinoma Treated with Transarterial 

Chemoembolization 

Neehar D. Parikh 1,3 , Amit G. Singal 2 , Peng Zhang 3 , Venkat Krishnamurthy 

3 , Pranab Barman 1 , Akbar K. Waljee 1,4 , Grace L. Su 1,3 ; 

1 Division of Gastroenterology, University of Michigan, Ann Arbor, 

MI; 2 Division of Digestive and Liver Diseases, Parkland Health and 

Hospital System, Dallas, TX; 3 VA Ann Arbor Healthcare System, 

Ann Arbor, MI; 4 Veterans Affairs Center for Clinical Management 

Research, VA Ann Arbor Healthcare System, Ann Arbor, MI 

The prognosis of patients with hepatocellular carcinoma (HCC) 

undergoing transarterial chemoembolization (TACE) is often 

uncertain. The existing HCC staging criteria are limited in their 

ability to predict survival after therapy. Analytic morphomics 

has been shown to predict patient outcomes in cirrhosis and 

other disease states. We aimed to determine the ability of 

analytic morphomics to predict outcome after TACE for HCC. 

We included patients from a single center (Ann Arbor VA Medical 

Center) who had TACE as the primary treatment for their 

HCC and who had a CT chest or abdomen prior to therapy. 

Analytic morphomic measurements were taken at the bottom of


the 11th thoracic vertebral level. We performed univariate and 

conditional inference tree analysis to identify the morphomic 

characteristics predictive of survival. Our primary outcome was 

actuarial survival at one year. We performed validation with 

patients who underwent TACE at two other centers (University 

of Michigan [n=85] and University of Texas Southwestern 

[n=72].) In total 76 patients (74 male; 2 female) were identified 

from the derivation cohort. Baseline characteristics, liver 

function and tumor stage are shown in the Table. Median survival 

was 564 days (95% CI: 416-713) from performance of 

the CT scan. Conditional inference tree analysis revealed that 

interstitial density (ITHU) was the only morphomic factor predictive 

of survival. Patients with ITHU above 0.709 had a 1-year 

survival of 51% (95% CI 43-59%) and those below 0.709 had 

a 1-year survival of 87% (95% CI: 81-93%) (p=0.001). External 

validation in two diverse external cohorts, showed that the 

ITHU cutoff was also predictive of one year survival in patients 

undergoing TACE at each site. In an exploratory analysis of the 

derivation cohort, we found that patients who had decompensation 

after TACE had a significantly higher ITHU than those 

who did not (p3 and FBG≥2.68g/L 

were independent risk factors of recurrence of HCC after LT. 

A scoring model was built to predict the risk of tumor recurrence, 

with a sensitivity of 68.3% and a specificity of 87.5%. 

Conclusion: Pretransplant elevated plasma fibrinogen level significantly 

increases the risk for tumor recurrence in patients 

after liver transplantation for HCC. The scoring model we built 

based on fibrinogen level and tumor number strongly correlates 

with tumor recurrence and may aid in the selection of patients 

that would most benefit from transplantation for HCC. 

Disclosures: 

The following authors have nothing to disclose: Guo-Ying Wang, Yang Yang, 

Gui-Hua Chen 

420 

Racial Disparity in Hepatocellular Carcinoma Patients 

Diagnosed in an Urban Medical Center 

Kirthi Lilley, Paul H. Naylor, Omar Sadiq, Sarah Stern, Sindhuri 

Benjaram, Karthik Ravindran, Samran Haider, Ryan Morton, Ravi 

Anand, Anupama Devara, Karan Mathur, Philip A. Philip, Murray 

N. Ehrinpreis, Milton G. Mutchnick; Internal Medicine, Wayne 

State University School of Medicine, Detroit, MI 

Background: Hepatocellular carcinoma (HCC) is a significant 

liver disease outcome and based on a previous report from 

our group, occurs in 23% of untreated African Americans (AA) 

with chronic hepatitis C (CHC) who were followed for an average 

of 8 years in an urban GI clinic. The aim of this study was 

to evaluate racial disparity with respect to the role of CHC and 

the development of HCC. Tumor size at first diagnosis and the 

role of surveillance prior to diagnosis of HCC were the primary 

foci of the study. Methods: The electronic medical records of 

the largest health care provider in Southeast Michigan and 

its associated multi-specialty group were used to identify all 

patients with HCC between 2012 and 2014. Using ICD-9 

codes 115.0 (hepatocellular carcinoma) and 115.2 (malignant 

neoplasm of liver, NOS), a patient pool of 281 potential 

HCC patients (67% AA) was identified. From the pool, 125 

patients were confirmed to have HCC by imaging (US /CT/ 

MRI) and/or biopsy. The remaining patients either had metastastic 

tumors or no confirmation of a HCC diagnosis. Race was 

determined by self-reporting, based primarily on skin color and 

was analyzed as AA (black) or Non-AA (non-black; Caucasian, 

Hispanic, Asian and other) Results: The 125 confirmed 

HCC patients were primarily AA (76%) and male (80%). The 

average age at diagnosis of HCC was 62 years (range for 

95% was 61-64 years). HCV was a dominant risk factor for 

the development of HCC in AA but not in non-AA (89% vs 

45%; p


the majority of patients were not diagnosed in a surveillance 

protocol. The majority of patients undergoing surveillance also 

had non-curable tumors at diagnosis. The optimal method for 

preventing HCC in AA remains effective treatment with the new 

anti-viral therapies which, unlike previous therapies are highly 

effective in AA. 

Disclosures: 

Milton G. Mutchnick - Advisory Committees or Review Panels: BMS; Grant/ 

Research Support: Janssen, Gilead; Speaking and Teaching: Janssen, Gilead, 

Abbvie, CLDF, Simply Speaking 

The following authors have nothing to disclose: Kirthi Lilley, Paul H. Naylor, 

Omar Sadiq, Sarah Stern, Sindhuri Benjaram, Karthik Ravindran, Samran 

Haider, Ryan Morton, Ravi Anand, Anupama Devara, Karan Mathur, Philip A. 

Philip, Murray N. Ehrinpreis 

421 

In patients with HCC and macrovascular invasion 

treated with sorafenib, AFP value≤1000 UI/L at baseline 

and vascular invasion not including 1 st order 

branches or main trunk (Vp1/2) are associated with 

prolonged survival. 

Charlotte E. Costentin 1 , Françoise Roudot-Thoraval 2 , Thomas 

Decaens 3 , Nathalie Ganne-Carrié 4 , Bernard Paule 5 , Eric Vibert 5 , 

Mélanie Chiaradia 6 , Christian Letoublon 7 , Julien Calderaro 8 , Giuliana 

Amaddeo 1 , Alexis Laurent 9 , Ivan Bricault 10 , Olivier Seror 11 , 

Didier Samuel 5 , Ariane Mallat 1 , Christophe Duvoux 1 ; 1 Hepatology, 

Hôpital Henri Mondor, Creteil, France; 2 Public Health, Henri 

Mondor Hospital, Creteil, France; 3 Hepato-gastroenterology, CHU 

Grenoble, Grenoble, France; 4 Hospital, Jean Verdier, Bondy, 

France; 5 Hepatology-hepatobiliary surgery, Paul Brousse Hospital, 

Villejuif, France; 6 Radiology, Henri Mondor Hospital, Creteil, 

France; 7 Surgery, CHU Grenoble, Grenoble, France; 8 Pathology, 

Henri mondor Hospital, Creteil, France; 9 Surgery, Henri Mondor 

Hospital, Creteil, France; 10 Radiology, CHU Grenoble, Grenoble, 

France; 11 Radiology, Jean Verdier Hospital, Bondy, France 

Introduction: According to EASL-OERTC guidelines, sorafenib 

is the standard of care for advanced hepatocellular carcinoma 

(HCC) BCLC-C although in the SHARP trial, macrovascular 

invasion (MVI) was associated with a poor prognosis. However, 

survival had been assessed for patients with MVI with 

or without extrahepatic spread (EHS) (8 months) but not in 

the group of patients with MVI but no EHS. The aim of this 

study was to describe outcome and predictors of survival 

in patients with HCC and MVI but no extra-hepatic spread 

treated with sorafenib. Methods: 67 patients with HCC and 

MVI but no EHS treated with sorafenib were identified in the 

database of 4 French centers (Bondy, Créteil, Grenoble,Villejuif) 

and retrospectively studied. Results: Patients (pts) were 

mostly males (88.1%), mean age 65.5±9.6 with underlying 

cirrhosis (89.4%). Etiology of liver disease was alcohol ±NASH 

(38.8%), viral hepatitis (35.8%), NASH without alcohol 

(14.9%), other (10.4%). Child-Pugh score was A and B in 59 

(86.4%) and 8 (13.6%) pts, respectively. HCC was uni-nodular 

in 17 pts (25.4%), infiltrative in 32 pts (47.8%), bilobar 

in 22 pts (33.3%). Median size of the larger nodule was 70 

[50-100] mm. MVI distribution was: Vp1+2 (invasion of or 

distal to the 2 nd order branches of the portal vein (PV)) in 8 

pts (12.0%), Vp3 (invasion of 1 st order branches of the PV) 

in 20 pts (30%), Vp4 (invasion of the main trunk and/or contra-lateral 

PV branch) in 32 pts (47.5%) and hepatic vein±Vp 

in 7 pts (10.5%). Median AFP level at baseline was 408 [32- 

2695] ng/ml. Median follow up was 6.7 [4.0-15.1] months. 

Median treatment duration on sorafenib was 6.4 months, and 

20 (30%) pts had additional therapies during follow up, either 

during sorafenib therapy or as second line (including TACE, 

systemic therapy, radiotherapy or radioembolization). Median 

overall survival was 12.6 months (IC95%: 8.4-16.7). In univariate 

analysis, AFP>1000 UI/L and 1 st order branches/troncular 

portal vascular invasion (Vp3-Vp4) were negatively associated 

with survival (p=0.049 and 0.031 respectively). In multivariate 

analysis, both parameters were independently associated 

with death: AFP>1000 (HR 2.188 [1.09-4.41], p=0.029), 

Vp3-Vp4 (HR 2.874[1.19-6.94], p=0.019). Median overall 

survival in pts with these 2 prognostic factors was 8.4 [4.2- 

12.5] months vs 16.9 [11.6-22.2] months in pts with one or 

none of these prognostic factors (p 1000 

UI/L and Vp3-Vp4 MVI are independent predictors of reduced 

survival. Long-term survival (median 16 months) was observed 

in pts with one or none of these 2 prognostic factors. 

Disclosures: 

Françoise Roudot-Thoraval - Advisory Committees or Review Panels: Roche; Consulting: 

LFB biomedicaments; Speaking and Teaching: gilead, Janssen, BMS, 

Roche 

Nathalie Ganne-Carrié - Advisory Committees or Review Panels: Roche; Speaking 

and Teaching: BMS, Gilead, Bayer 

Olivier Seror - Board Membership: Bayer Healthcare; Consulting: Olympus; 

Speaking and Teaching: Angiodynamics 

Didier Samuel - Consulting: Astellas, MSD, BMS, Roche, Novartis, Gilead, LFB, 

Janssen-Cilag, Biotest, Abbvie 




The following authors have nothing to disclose: Charlotte E. Costentin, Thomas 

Decaens, Bernard Paule, Eric Vibert, Mélanie Chiaradia, Christian Letoublon, 

Julien Calderaro, Giuliana Amaddeo, Alexis Laurent, Ivan Bricault, Ariane Mallat 

422 

Lamivudine vs. Entecavir for the newly diagnosed hepatitis 

B-virus related Hepatocellular carcinoma 

jung hee kim, Dong Hyun Sinn, Kyunga Kim, Geum-Youn Gwak, 

Yong-Han Paik, Moon Seok Choi, Joon Hyeok Lee, Kwang Cheol 

Koh, Seung Woon Paik; samsung medical center, Seoul, Korea 

(the Republic of) 

Antiviral therapy is a key element in the management of hepatitis 

B virus (HBV)-related hepatocellelular carcinoma (HCC) 

patients. However, little is known whether potent drug, entecavir, 

is more effective than a less potent drug, lamivudine, 

in reducing the risk of death, hepatic decompensation and 

recurrence in HBV-related HCC. A retrospective cohort of 451 

newly-diagnosed, HBV-related HCC patients without antiviral 

therapy at diagnosis, and started antiviral therapy with either 

entecavir (n = 249) or lamivudine (n=202) were enrolled. 

Overall survival, new-onset hepatic decompensation (ascites, 

variceal bleeding, hepatic encephalopathy), and recurrence 

after curative therapy were compared. Baseline HBV DNA levels 

were not diffference between two groups but rescue therapy 

was more frequent in lamvudine group.(45% vs. 6.4%, 

p


vious hepatic decompensation, and was also independent risk 

factor for recurrence after curative therapy [HR (95% CI): 1.84 

(1.25-2.72), p = 0.002] among 117 patients who received 

curative therapy. The overall survival, decompensation-free 

survival, and recurrence-free survival was better in entecavir 

treated patients than lamivudine treated patients . The survival 

benefit of choosing high potent drug over less potent drug will 

be higher when patient survival is long enough. Low-potent 

drug may be a valuable option for those with low expected 

survival, especially in resource-limited setting, but high-potent 

drug should be the preferred choice in the rest of HBV-related 

HCC patients. 

Disclosures: 

The following authors have nothing to disclose: jung hee kim, Dong Hyun Sinn, 

Kyunga Kim, Geum-Youn Gwak, Yong-Han Paik, Moon Seok Choi, Joon Hyeok 

Lee, Kwang Cheol Koh, Seung Woon Paik 

423 

Long term outcome after resection of intermediate 

hepatocellular carcinoma: comparison to transarterial 

chemoembolization 

DAEGEON AHN, Dong Hyun Sinn, Geum-Youn Gwak, Moon 

Seok Choi, Joon Hyeok Lee, Kwang Cheol Koh, Seung Woon 

Paik, Byung Chul Yoo; Gastroenterology, Samsung Medical Center, 

Seoul, Korea (the Republic of) 

Background and Aims: Transarterial chemoembolization 

(TACE) is the first-line recommended therapy for patients with 

intermediate hepatocellular carcinoma (HCC) in the BCLC staging 

system. However, in clinical practice, some patients receive 

hepatectomy. We compared long-term outcome in these 

patients. Methods: A total of 277 patients with intermediate 

stage HCC who were treated with either hepatectomy or TACE 

were analyzed. Results: The median survival was significantly 

better for resection than TACE (5.3 vs. 2.6 years, p = 0.002), 

however, when adjusted for age, Child-Pugh Class, maximal 

tumor size, tumor number, uni- or bi-lobal involvement, and AFP 

levels, there was no significant survival difference by treatment 

modality [Hazard ratio (HR): 1.42, 95% CI (0.91-2.22), p = 

0.11]. In sub-group analysis, resection showed better survival 

than TACE in Child-Pugh score 5, maximal tumor size ≤ 5cm 

or > 10cm, and tumor number ≤ 3, however, there was no 

survival difference in patients with Child-Pugh score ≥ 6, tumor 

size between 5-10cm, and tumor number ≥ 4. When patients 

were sub-grouped according to the Child-Pugh Class, tumor 

number and size, resection (n = 26) provided survival benefit 

over TACE (n = 58) for those with Child-Pugh class 5, less than 

3 tumors, and tumor size ≤ 5cm or > 10cm [adjusted HR: 0.41 

(95% CI: 0.20-0.84), p = 0.015], but not the rest of patients 

[adjusted HR for resection (n = 26) vs. TACE (n = 167): 0.73 

(95% CI: 0.44-1.21), p = 0.22]. Conclusion: Resection can 

be first-line treatment option for intermediate stage HCC when 

certain criteria are met. In this study, child-Pugh score 5, tumor 

number ≤ 3, and tumor size ≤ 5 or > 10 cm were the criteria. 

Disclosures: 

The following authors have nothing to disclose: DAEGEON AHN, Dong Hyun 

Sinn, Geum-Youn Gwak, Moon Seok Choi, Joon Hyeok Lee, Kwang Cheol Koh, 

Seung Woon Paik, Byung Chul Yoo 

424 

TBI 302: A first-in-class therapy for the treatment of 

advanced stage liver cancer 

Steve Brookes 1 , Gary Levy 1,2 , Jin Seog Seo 1 , Murray J. Cutler 1 , 

Yvonne Frater 1 , Gord Adamson 1 , David Bell 1 ; 1 Drug Development, 

Therapure Biopharma Inc., Mississauga, ON, Canada; 2 Multi 

Orgon Transplant Program, University of Toronto Transplant Institute, 


Hepatocellular carcinoma (HCC) is the second most common 

cause of death from cancer worldwide; few treatment options 

are available, especially for advanced stage disease. Current 

standard of care provides only a modest improvement in overall 

survival and is associated with poor quality of life. Therapure 

has developed a novel drug delivery platform based upon 

the attachment of therapeutic drugs to hemoglobin (Hb) as a 

means of targeting the liver. TBI 302 is a hemoglobin-drug conjugate 

(HDC) designed to deliver the anticancer drug floxuridine 

(FUdR) to the liver to improve the effectiveness of treatment 

for HCC. Systemic toxicity associated with free FUdR restricts its 

use to locoregional administration (0.2-0.5 mg/kg/d) via continuous 

hepatic arterial infusion (HAI). Although HAI of FUdR 

can reduce hepatic tumor burden, toxicity from HAI FUdR and 

complications associated with direct hepatic infusion pumps 

can be significant. HDC technology uses Hb as an innovative 

drug carrier that exploits the natural pathway for hemoglobin 

clearance predominantly through the liver to provide selective 

drug targeting while preserving FUdR activity following standard 

intravenous (IV) infusion. To establish a safe starting dose 

for a first-in-human Phase 1 trial, a GLP-compliant repeat dose 

preclinical safety study of TBI 302 in cynomolgus monkeys was 

conducted. TBI 302 administered by 1-hour IV infusion once 

per week for 8 weeks at doses of 2, 5, and 10.5 mg/kg (0.08, 

0.19, and 0.4 mg/kg FUdR, respectively) was well tolerated 

at all dose levels. Increasing doses of TBI 302 resulted in proportional 

increases in area under the concentration-time curve 

(AUC) and maximum concentration (Cmax) of total plasma 

FUdR. No clinical signs or biochemical toxicity was associated 

with IV infusion of TBI 302. The no-observed-adverse-effect 

level (NOAEL) of TBI 302 was determined to be the highest 

dose level of 10.5 mg/kg (0.4 mg/kg FUdR). TBI 302 is projected 

to have a half-life of approximately 5 hours in humans. A 

Phase I safety study of TBI 302 as second-line therapy in HCC 

has been approved by the US FDA. The primary objective is 

to determine safety and tolerability of TBI 302. The secondary 

objectives are to determine TBI 302 pharmacokinetics and 

effects on tumor burden. Therapure’s HDC platform represents 

a new class of therapy that offers liver-specific targeting for 

a wide range of hepatic diseases, while potentially reducing 

extra-hepatic toxicity of drugs. 

Disclosures: 

Jin Seog Seo - Employment: Therapure Biopharma Inc 

Murray J. Cutler - Employment: Therapure Biopharma 

Yvonne Frater - Management Position: Therapure 

Gord Adamson - Employment: Therapure Biopharma Inc. 

David Bell - Employment: Therapure Biopharma Inc. 

The following authors have nothing to disclose: Steve Brookes, Gary Levy


425 

Staging Hepatocellular Carcinoma in Non-Cirrhotic 

Patients 

Philippe Kolly 1,2 , Helen Reeves 3,4 , Marina Knöpfli 2 , Jean-Francois 

Dufour 1,2 ; 1 Hepatology, Department of Clinical Research, University 

of Bern, Bern, Switzerland; 2 University Clinic for Visceral 

Surgery and Medicine, Inselspital Bern, Bern, Switzerland; 3 The 

Northern Institute for Cancer Research, Newcastle University, 

Newcastle-upon-Tyne, United Kingdom; 4 The Liver Unit, Freeman 

Hospital, Newcastle-upon-Tyne, United Kingdom 

To design a staging system for HCC in non-cirrhotic patients, 

we used a pooled set of 864 patients combining 2 prospective 

European cohorts. Among them, 221 had a non-cirrhotic 

liver. Cirrhosis was excluded based on clinical presentation 

and imaging. The staging and treatment algorithm was constructed 

by using Cox regression, regression tree analysis and 

evidenced based literature. We created the Non-Cirrhotic Liver 

Cancer (NCLC) staging system. Based on the ECOG PS, tumor 

burden (size and number of nodules), presence of vascular 

invasion and metastasis, this staging system proposes 4 stages. 

Each stage is associated with treatment options, separated in 

first and second intention treatments (Fig.). We assessed the 

discriminatory power of the staging system by Log-Rank (p 

< 0.001). The prognostic ability of the staging system was 

assessed with Harrell’s concordance index (0.729). HCC 

developing in non-cirrhotic patients represents 10-40% of 

HCC. With the growing incidence of NAFLD-associated HCC, 

this proportion will rise. The BCLC staging system, which has 

been endorsed by AASLD and EASL, has not been developed 

for non-cirrhotic patients. The statistical analysis revealed that 

the performance status and the tumor burden are discriminating 

parameters to classify non-cirrhotic patients with HCC in 4 

different stages with different prognostics and treatment allocation. 

A staging system for HCC developing in non-cirrhotic 

patients has become essential. The NCLC has been designed 

specifically for this purpose and will need to be further developed. 

Figure legend: NCLC staging and treatment algorithm. 

This algorithm stages HCC in non-cirrhotic patients according 

to ECOG PS, tumor burden and the existence of vascular invasion 

or metastasis. It suggests 4 stages and the corresponding 

treatments, separated in first and second intention. 

426 

Clinical significance of microRNA-31 expression in 

human hepatocellular carcinoma 

Keun Hur 1 , Se Young Jang 2 , Soo Young Park 2 , Gyeonghwa Kim 1 , 

Yong-Hun Choi 1 , Yu Rim Lee 2 , Su Hyun Lee 2 , Sun Kyung Jang 2 , 

Won Young Tak 2 , Young Oh Kweon 2 ; 1 Department of Biochemistry 

and Cell Biology, Kyungpook National University School of 

Medicine, Daegu, Korea (the Republic of); 2 Department of Internal 

Medicine, Kyungpook National University Hospital, Daegu, Korea 


Background & aims Hepatocellular carcinoma (HCC) is one 

of the most leading cause of cancer-related death worldwide. 

It is therefore important to understand the mechanistic roles of 

biomolecules involved in HCC development. MicroRNAs (miR- 

NAs) are small noncoding RNAs that act as an important regulator 

of gene expression in various tumor development as well 

as metabolic diseases. This study aim to investigate the role 

of miR-31 in HCC and non-alcoholic steatohepatitis (NASH). 

Methods We analyzed clinical specimens from HCC, matched 

normal liver, and NASH tissues. MicroRNAs expression levels 

were evaluated by quantitative real-time PCR (qRT-PCR) and 

miR-31 expression was normalized relative to RNU6B expression. 

In addition, we determined the clinical significance of 

the miR-31 expression in matching tissue and serum samples 

from HCC patients. Results MiR-31 expression was significantly 

upregulated in HCC and NASH tissues compared with normal 

liver tissues (p=0.0015). In HCC patients, lower level of 

miR-31 (p


Pearson chi squared tests we examined the rates of false-positive 

HCC by UNOS region and transplant center. Results: 

Among 3,852 transplant recipients with T2MELD exceptions 

and available explant data, 262 (6.8%) had no evidence of 

HCC identified on explant pathology. Rates of false-positive 

HCC were higher for patients who underwent prior HCC treatment, 

10.1% vs 6.5%, p = 0.014. Rates of false-positive HCC 

varied by UNOS region: 2.1% false-positive rate in region 6 

versus 12.0%% in region 7 (p = 0.001). There was significant 

among-center variability (Figure) in false-positive rates among 

those centers who transplanted ≥30 recipients with T2MELD 

exceptions during the study period: ranging from 0% in four 

centers to 46.0%at one center, p90%, even in patients with advanced 

hepatic fibrosis. On the other hand, hepatocellular carcinoma 

(HCC) develops in up to 4% of patients after successful treatment 

of continuous HCV infection using interferon therapy. It is 

important to evaluate hepatic pathology after HCV eradication. 

The aim of the present study was to characterize microRNA 

(miRNA) expression in the liver tissue of patients who achieved 

an SVR. Patients and Methods: 71 tissue samples from patients 

who underwent HCC resection were evaluated in the present 

study: 61 HCCs from patients with continuously infected HCV 

(HCV-HCC) and 10 from patients who had achieved SVR (SVR-


HCC). We also included non-tumorous tissues (SVR-NT) from 

four patients with SVR-HCC and liver tissue (SVR-CH) from four 

patients with SVR but without HCC. Total RNA was extracted 

from the liver samples. The miRNA expression patterns were 

analyzed using microarray. Using leave-one-out cross-validation, 

we picked up several miRNAs to classify samples between 

SVR-HCC and HCV-HCC and between SVR-NT and SVR-CH. In 

addition, target gene expression was quantified after miRNA 

overexpression in HEK293 cells. Results: We were able to 

discriminate between SVR-HCC and HCV-HCC with 75.36% 

accuracy using the expression pattern of six specific miRNAs 

(let-7a, let-7f, miR-21, miR-122, miR-720, and miR-923). The 

expression levels of 37 miRNAs were significantly lower in 

HCV-HCC than in SVR-HCC, whereas the expression of 25 

miRNAs was significantly higher in HCV-HCC than in SVR- 

HCC (P < 1.0e-05). The expression of thrombospondin 1 was 

regulated in an opposing manner by miR-30a-3p in SVR-HCC 

and HCV-HCC. In non-tumorous tissues, the expression pattern 

of seven miRNAs distinguished between SVR-CH and SVR-NT 

with 87.50% accuracy. Conclusions: Comprehensive miRNA 

expression analyses can not only differentiate between SVR- 

HCC and HCV-HCC, but can also predict hepatocarcinogenesis 

after achieving an SVR. 

Determining SVR-HCC using 6 miRNAs (let-7a, let-7f, miR-21, 

miR-122, miR-720, and miR-923) expression pattern in LOOCV 

analysis. 

The column and row showed prediction and result, respectively. 

For example in 5 years observation, 47 examples of SVR-HCC 

expected as HCV-HCC, and it were 3 examples in HCV-HCC. 

The prediction accuracy between HCV-HCC and SVR-HCC-5 

years was 75.36%. 

Disclosures: 

Norifumi Kawada - Grant/Research Support: BMS, Chugai, Kowa; Speaking 

and Teaching: MSD, Janssen 

The following authors have nothing to disclose: Akihiro Tamori, Yoshiki 

Murakami, Shoji Kubo, Saori Itami, Sawako K. Uchida, Hiroyasu Morikawa, 

Masaru Enomoto, Shigekazu Takemura, Toshihito Tanahashi, Y-h Taguchi 

430 

Exploration of cancer-prone gut microbiota in chronic 

hepatitis C patients 

Noriho Iida, Eishiro Mizukoshi, Shuichi Kaneko; Kanazawa Univ, 

Kanazawa, Japan 

Purpose: Recently it has been recognized from animal studies 

that dysbiosis of gut microbiota is one of the causes of liver 

carcinogenesis. However, character of gut microbiota prone 

to liver carcinogenesis in human patients remains unclear. In 

this study we aimed to characterize gut microbiota of chronic 

hepatitis C patients with hepatocellular cancers (HCC) and 

determine whether the microbiota derived from HCC patients 

actually cause liver carcinogenesis in mice or not. Methods: 

Feces were collected from 11 chronic hepatitis C patients with 

primary HCC, 8 chronic hepatitis C patients without HCC 

(CLD), and 6 healthy donors (HD). After DNA was extracted 

from the fecal samples, libraries were generated with Illumina 

Nextera DNA kits to do whole genome shotgun sequencing. 

The sequencing was performed with Illumina Miseq. After quality 

control, removal of human genome sequences and PCR 

replicative sequences, the sequences were analyzed with 

MetaPhLAN or HUMAnN, to know taxonomic profile or presence 

of genes, respectively. To find significantly differential 

bacteria species or genes, linear discriminative analyses (LDA) 

were done. To test causality of the gut microbiota derived from 

HCC patients on liver carcinogenesis, fecal suspension made 

from the patients or HD was transplanted to C57BL6 mice in 

which gut bacteria were depleted by oral antibiotic cocktail. 

The human feces-transplanted mice were injected with diethylnitrosamine 

(DEN) and CCl4 to induce liver tumors. Results: 

Total 57,259,209 sequences were analyzed on MetaPhLAN 

and HUMAnN. Alpha-Diversity in the sample (Simpson’s index) 

was not significantly different among the groups. HCC group 

was not differentially clustered from HD group. However, in 

the comparison of HCC vs HD on LDA analysis, 8 taxa were 

significantly increased and 2 taxonomy decreased in HCC 

(P


non-HCC liver tissue in relation to RNU6B. Results: Considering 

all 45 HCC patients, lower levels of hsa-miR- 125a-5p were 

observed in HCC tissue than in non-HCC liver tissues (M+SD 

4.84+3.69 vs. 8.27+4.5 A.U., p=0.0002). This difference 

was highly significant to statistical analysis in the 35 HCV-patients, 

4.21+ 3.47 vs. 7.79+4.75 AU, p=0.0009 and lower 

and not statistically significant significance in the 5 HBV-patients, 

7.38+2.08, vs. 10.7+2.4AU, and. in the 5 patients 

with NASH-related cirrhosis, 6.72+5.24 vs. 9.2+3.99AU. In 

addition, the level of miR-125a-5p in the HCC tissue of the 35 

HCV-related cirrhosis was lower than that observed in the 10 

non-HCV patients (4.21+3.47 vs. 7.05+3.77 AU, p=0.023). 

Besides, the levels of has-miR-125a-5p in HCC tissue were 

moderately lower in the 41 patients with Child-Pugh score A 

(4.67+3.47 AU) than in the 4 with score B (7.05+3.77 AU, 

p=0.7), such as in the 40 patients with BCLC class A as compared 

with the 5 with class B or C (4.66+3.78 vs. 6.31+2.79 

AU, p=0.21). Conclusions: These data suggest an oncosuppressor 

effect of microRNA hsa-miR-125a-5p on HCV-related 

HCC and possibly even in non-HCV HCC,, an observation 

deserving further investigation 

Disclosures: 

The following authors have nothing to disclose: Nicola Coppola, Giorgio de 

Stefano, Nicola Mosca, Valentina Iodice, Carmine Minichini, Filomena Castiello, 

Nunzia Farella, Mario Starace, Nicoletta Potenza, Giulia Liorre, Lorenzo Onorato, 

Evangelista Sagnelli, Aniello Russo 

432 

Surgical resection versus radiofrequency ablation plus 

drug-eluting beads transcatheter arterial chemoembolization 

for the treatment of solitary large hepatocellular 

carcinoma. A single center experience 

Antonio Saviano 1 , Roberto Iezzi 2 , Felice Giuliante 3 , Lucia Salvatore 

1 , Caterina Mele 3 , Alessandro Posa 2 , Mariachiara Campanale 

1 , Emanuele Rinninella 1 , Valentina Cesario 1 , Federico 

Barbaro 1 , Marco Biolato 1 , Maria Assunta Zocco 1 , Laura Riccardi 1 , 

Brigida E. Annicchiarico 1 , Massimo Siciliano 1 , Anna Maria De 

Gaetano 2 , Antonio Grieco 1 , Gian Ludovico Rapaccini 1 , Antonio 

Gasbarrini 1 , Lorenzo Bonomo 2 , Maurizio Pompili 1 ; 1 Department 

of Internal Medicine, Catholic University, Rome, Italy; 2 Department 

of Bioimaging and Radiological Sciences, Institute of Radiology, 

Catholic University, Rome, Italy; 3 Hepatobiliary Surgery Unit, 

Department of Surgery, Catholic University, Rome, Italy 

Background and aims: Aim of this study was to compare 

liver resection (RES) and single-step combined therapy with 

radiofrequency ablation and drug-eluting beads transarterial 

chemoembolization (RFA+TACE) in patients with single hepatocellular 

carcinoma (HCC) larger than 3 cm and compensated 

cirrhosis. The primary end-points were overall survival (OS) 

and tumour recurrence (TR) rates. Methods: This retrospective 

study involved 56 Child-Pugh A cirrhotic patients (25 in RES 

group and 33 in RFA +TACE group) with single HCC observed 

between 2010 and 2014 in our Center. All the patients did 

not show macroscopic vascular invasion and/or extrahepatic 

tumor spreading before treatment. In all cases the treatment 

choice was made after multidisciplinary evaluation. RFA and 

TACE were performed using a single step procedure (RFA 

during balloon artery occlusion of the tumour feeding artery 

followed by TACE) . The cumulative OS and TR rates were analysed 

using the Kaplan Meyer method. Results: The median age 

and follow up period of the whole population were, respectively, 

70 yrs and 16.5 months. The median HCC size was 

4,5 cm in RES group (range 3-7.4 cm) and 4 cm in RFA+TACE 

group (range 3-6.8 cm) (p 0.150). The tumours larger than 

5 cm were 40.0% in RES group and 21.9% in RFA group (p 

0.138). Demographic, clinical and laboratory parameters did 

not differ significantly between groups but RFA+TACE patients 

showed clinically evident portal hypertension in a higher rate 

of cases (32.0% in RES group, 63.6% in RFA+TACE group, p 

0.017).Treatment-related complications did not differ significantly 

between groups. OS rates at 1-3 years were 91.8% 

- 75.2% in RES group and 86.3% - 61.6% in RFA+TACE group 

(p 0.463). Overall TR rates at 1-3 years were significantly 

lower in resected patients (RES group: 32.7% - 42.3%; RFA+- 

TACE group 43% - 74.7%, p 0.039). Both 3-year local tumour 

progression (RES group 31.1%, RFA+TACE group 74.6%, p 

0.004) and distant intrahepatic recurrence (RES group 34.8%, 

RFA+TACE group 77.0%, p 0.043) occurred significantly more 

frequently in RFA+TACE group. Considering only patients with 

HCC ≤ 5 cm, the OS rates at 1-3 years were 100.0% - 84.6% 

in RES group and 87.1% - 62.3% in RFA+TACE group (p 

0.218). Furthermore, TR rates at 1-3 years were significantly 

lower in RES group than in RFA+TACE group (20.6% - 33.8% 

versus 45.3% - 65.8%, p 0.049). Conclusions: RES appears 

to be the choice treatment in compensated cirrhotic patients 

with single HCC larger than 3 cm. One step combined RFA+- 

TACE treatment is an effective alternative therapeutic option in 

patients who are not candidates to RES after multidisciplinary 

evaluation. 

Disclosures: 

The following authors have nothing to disclose: Antonio Saviano, Roberto Iezzi, 

Felice Giuliante, Lucia Salvatore, Caterina Mele, Alessandro Posa, Mariachiara 

Campanale, Emanuele Rinninella, Valentina Cesario, Federico Barbaro, 

Marco Biolato, Maria Assunta Zocco, Laura Riccardi, Brigida E. Annicchiarico, 

Massimo Siciliano, Anna Maria De Gaetano, Antonio Grieco, Gian Ludovico 

Rapaccini, Antonio Gasbarrini, Lorenzo Bonomo, Maurizio Pompili 

433 

Efficacy of Radiotherapy in addition to Chemoembolization 

and Hepatic Arterial Infusion Chemotherapy versus 

Sorafenib in Advanced Hepatocellular Carcinoma with 

Portal Vein Thrombosis 

Sun Young Yim 1 , Soon Ho Um 1 , Tae Hyung Kim 1 , Jem Ma Ahn 1 , 

Beom Jin Park 2 , Yeon Seok Seo 1 , Ho Sang Ryu 1 ; 1 Department of 

Internal Medicine, Korea University College of Medicine, Seoul, 

Korea (the Republic of); 2 Radiology, Korea University College of 


Background and aim: Sorafenib is the only standard treatment 

for advanced hepatocellular carcinoma with (HCC). However, 

its efficacy is not satisfactory and other treatment options are 

required. This study investigated the efficacy of chemoembolization 

and hepatic arterial infusion chemotherapy (HAIC) 

with or without radiotherapy versus sorafenib alone. Methods: 

This single-center retrospective study involved 105 patients 

of advanced HCC with portal vein tumor thrombosis (PVT). 

Enrolled patients had either child-pugh (CP) class A or B liver 

cirrhosis whom were classified into 3 groups: 1) Sorafenib 

alone, n=20; 2) Chemoembolization and HAIC, n=26; 3) 

Chemoembolization and HAIC with radiotherapy, n=59. 

Sorafenib was initiated with 400mg twice daily and HAIC 

was based on cisplatin and 5-fluorouracil regimen, performed 

every 4 weeks. Response of PVT was determined 3 months 

after completion of treatment and was regarded as responsive 

when there is at least partial response. Overall survival (OS) 

was analyzed among the treatment groups and factors associated 

with mortality were evaluated using multivariate analysis. 

Result: The median radiation dose, sorafenib treatment duration 

and chemoembolization sessions were 50 Gy, 40 days, 

and 4 sessions, respectively. Proportion of patients according 

to the degree of PVT (main/both vs. first order vs. segmental 

PV) and bilobar tumor mass involvement did not differ among 

the three groups of treatment. However, PVT response rate


was significantly higher in group 3 (13.5% vs. 6.7% vs. 55%, 

P=0.014) with lower incidence of solid organ metastasis (60% 

vs. 23.1% vs. 18.6%, P=0.001) and CP class B (70%, 50%, 

25.4%%, P=0.001) compared to group 2 and 1. In univariate 

cox analysis, treatment modalities, presence of either lymph 

node or other organ metastasis, CP class B, and decrease in 

AFP levels were associated with survival. However, multivariate 

analysis revealed that treatment modalities (group 1 vs. 

2, HR, 0.244; 95% CI, 0.06-0.999, P=0.05, group 1 vs. 3, 

HR, 0.121; 95% CI 0.028-0.51, P=0.004 and group 2 vs. 

3, HR 0.495; 95% CI, 0.25-0.98, P=0.044) and decrease in 

serum AFP level within 2 months of treatment (HR, 1.813; 95% 

CI, 1.204-2.72; P=0.004) were the only independent factors 

associated with survival. Median OS was significantly higher 

in patient treated with radiotherapy (group3, 11.1 months) 

than group 2 (3.6 months, log rank P


Disclosures: 

The following authors have nothing to disclose: Andrea Casadei Gardini, Giorgia 

Marisi, Paola Ulivi, Luca Faloppi, Emanuela Scarpi, Mario Scartozzi, Francesco 

G. Foschi, Massimo Iavarone, Gianfranco Lauletta, Jody Corbelli, Elena 

Tenti, Floriana Facchetti, Cristina Della Corte, Stefano Tamberi, Oriana Nanni, 

Stefanu Cascinu, Dino Amadori, Giovanni Luca Frassineti 

other liver diseases (4%). There was no significant difference in 

frequency of miR-26a loss in patients with advanced versus less 

advanced HCC regardless of treatment (79% vs 74% in resection 

cases and 50% vs 60% in transplanted cases). Conclusions: 

Loss of miR-26a in the tumor tissue relative to surrounding 

liver tissue is a common phenomenon in HCC regardless the 

etiology or prior liver directed therapy but it was not associated 

with an aggressive tumor phenotype. More studies are needed 

to understand the biological significance of this observation. 

Disclosures: 



The following authors have nothing to disclose: Sarah C. Nabinger, Samer Gawrieh, 

Maaz B. Badshah, Sandra K. Althouse, Smitha Marri, Sangbin Lee, Smiti S. 

Sahu, Susan Perkins, Romil Saxena, Janaiah Kota 

436 

MicroRNA-26 expression is significantly lower in hepatocellular 

carcinoma relative to surrounding liver tissue 

in humans with chronic liver disease 

Sarah C. Nabinger 1 , Samer Gawrieh 3 , Maaz B. Badshah 3 , Sandra 

K. Althouse 2 , Smitha Marri 3 , Sangbin Lee 1 , Smiti S. Sahu 1 , 

Susan Perkins 2 , Romil Saxena 4 , Naga P. Chalasani 3 , Janaiah 

Kota 1 ; 1 Medical and Molecular Genetics, Indiana University 

School of Medicine, Indianapolis, IN; 2 Biostatistics, Indiana University 

School of Medicine, Indianapolis, IN; 3 of Gastroenterology/ 

Hepatology, Indiana University School of Medicine, Indianapolis, 

IN; 4 Pathology and Laboratory Medicine, Indiana University 

School of Medicine, Indianapolis, IN 

Background & Aim: Hepatocellular Carcinoma (HCC) is the 

third leading cause of cancer-related deaths worldwide. We 

have previously shown that loss of microRNA-26a (miR-26) 

is common in murine HCC and systemic delivery of miR-26a 

suppressed liver tumor development in a mouse model of HCC. 

In this study, we investigated if miR-26 loss occurs with HCC 

in a large cohort of patients with underlying liver disease and 

assessed its correlation with advanced tumor features. Methods: 

We identified 136 paired tumor and normal adjacent 

liver tissues from HCC patients undergoing tumor resection 

(n=87) or liver transplantation (n=49) at Indiana University 

Medical Center. Total RNA was isolated from formalin fixed 

paraffin embedded tumor and normal adjacent liver samples, 

and subjected to quantitative PCR analysis to estimate miR-26a 

levels in these patients. The prevalence of miR-26 loss, defined 

by a relative fold change


438 

Sorafenib Therapy in HIV-infected Patients with Hepatocellular 

Carcinoma (HCC) 

Luciana Kikuchi 1 , Caitlin C. Citti 2,3 , Ting-Yi Chen 4 , Heather L. 

Platt 3 , David E. Kaplan 5 , Meritxell Ventura-Cots 6 , Mamta K. Jain 4 , 

Eugenia Vispo 7 , Jorge Daruich 8 , Marina Nunez 9 , Pablo Barreiro 7 , 

Beatriz Minguez 6 , Ayse Aytaman 10 , David Rimland 11 , Norbert 

Bräu 3,2 ; 1 Gastroenterology, Universidade de São Paulo, São 

Paulo, Brazil; 2 Infectious Diseases and Liver Diseases, Icahn School 

of Medicine at Mount Sinai, New York, NY; 3 James J. Peters VA 

Medical Center, Bronx, NY; 4 University of Texas Southwestern 

Medical Center, Dallas, TX; 5 University of Pennsylvania, Philadelphia, 

PA; 6 Hospital Universitario Vall d’Hebron, Barcelona, Spain; 

7 Hospital Carlos III, Madrid, Spain; 8 Universidad de Buenos Aires, 

Buenos Aires, Argentina; 9 Wake Forest University, Winston-Salem, 

NC; 10 VA New York Harbor HCS, Brooklyn, NY; 11 Atlanta VA 

Medical Center, Atlanta, GA 

BACKGROUND: Sorafenib is the only systemic chemotherapy 

for HCC shown to decrease all-cause mortality. Little is known 

about its effect in HIV-infected patients with HCC. METHODS: 

HIV-infected patients with HCC who received sorafenib (with or 

without TACE) were retrospectively identified from 1992-2013 

in 38 centers in 8 countries. They were compared to HIV-infected 

patients from the same cohort who received no effective 

HCC therapy. RESULTS: Compared to 132 untreated patients, 

the 22 patients receiving sorafenib were of similar age (51 vs. 

53 years), but tended to have less frequently alcohol abuse 

(18% vs. 38%, p=0.079), were diagnosed through HCC 

screening more often (59 vs. 30%, p=0.009), and had lower 

mean Child-Pugh scores (6.6 vs. 7.8, p=0.001). Sorafenib 

patients had lower mean HIV viral loads (1.7 vs. 2.7 log10 

copies/ml, p


dictors of RFS in overall patients include cirrhosis (hazard ratio 

(HR)=2.337, p=0.006) and AFP level >20 ng/mL (HR=1.896, 

p=0.026). The median RFS in NUC secondary prevention failure 

and tertiary prevention groups were 54 and 36 months, 

respectively (p=0.467). The 1-, 2- and 5-year RFS rates were 

85.3%, 72.7%, 46.1% in the NUC secondary prevention failure 

group, and 76.4%, 61.3%, 46.6% in tertiary prevention 

group, respectively. Baseline viral loads, HBsAg levels and 

virological response were not associated with RFS. In the subgroup 

patients without cirrhosis, NUC secondary prevention 

failure group had a trend of better RFS (p=0.057), whereas 

the RFS was comparable between the two group in patients 

with cirrhosis. Conclusions Continue NUC treatment has the 

potential to reduce the risk of HCC recurrence in non-cirrhotic 

patients despite secondary prevention had failed. Closely monitoring 

is recommended for HBV-HCC patients with cirrhosis 

after curative surgical resection even under NUC treatment. 

Disclosures: 

The following authors have nothing to disclose: I-Cheng Lee, Yi-Hsiang Huang, 

Gar-Yang Chau, Teh-Ia Huo, Chien-Wei Su, Han-Chieh Lin 

440 

A Pilot Safety Study of Nature Killer Cells From Sibship 

to Treat the Recurrence of Hepatocellular Carcinoma 

After Liver Transplantation 

Guo-Ying Wang, Yang Yang, Gui-Hua Chen; Liver Transplantation 

Center, the Third Affiliated Hospital, Sun Yat-sen University, Organ 

Transplantation Research Institute, Guangzhou, China 

Objective: Nature Killer (NK) Cells have been thought to play 

a pivotal role in innate immunity. However, the safety and efficacy 

of NK cells for the treatment of hepatocellular carcinoma 

(HCC) recurrence after liver transplantation (LT) is unknown. In 

this study, we investigated whether the injection of activated 

NK cells (CD3 - CD56 + cells) from the sibship with the same 

blood type is safe for the treatment of HCC recurrence after LT. 

Methods: Six patients with HCC recurrence after LT were eligible 

and enrolled in this study. The patients received injections 

of 5 × 10 9 NK cells from the sibship with the same blood type 

4 times at a frequency of once every two weeks. Lymphocytes 

were extracted from the peripheral blood mononuclear cells 

and cultured with IL-2 and other cytokines for 2 weeks. The 

purity of lymphocytes was assessed by flow cytometric analysis, 

and only the CD3 - CD56 + cells greater than 70% were 

used. The adverse effects, laboratory tests, overall survival 

were assessed. Results: No serious adverse effects, laboratory 

abnormalities were identified as related to the treatment of 

NK cells. Fix patients were alive in the follow-up period of 26 

months while one died of liver failure due to tumor progression 

after 2 months of NK cells infusion. There was no graft-versushost 

disease in all 6 patients during follow-up. Conclusion: 

In conclusion, we have demonstrated for the first time to our 

knowledge that NK cells from sibship with the same blood type 

can be used safely as adoptive immunotherapy for the treatment 

of HCC recurrence after liver transplantation. 

Disclosures: 

The following authors have nothing to disclose: Guo-Ying Wang, Yang Yang, 

Gui-Hua Chen 

441 

Serological markers of extracelllular matrix remodelling 

for the diagnosis of HCC and the evaluation of tumour 

pathogenesis 

Roslyn Vongsuvanh 1 , Jacob George 1 , David van der Poorten 1 , 

Morten A. Karsdal 2 , Diana J. Leeming 2 ; 1 Storr Liver Unit, Westmead 

Millenium Institute, Westmead Hospital, Westmead, NSW, 

Australia; 2 Nordic Bioscience, Fibrosis Biology and Biomarkers, 

Herlev, Denmark 

Liver fibrosis is associated with HCC, however, whether markers 

of matrix turnover may be exploited for HCC evaluation, 

remains poorly understood. Fragments of the extracellular 

matrix (ECM) are released systemically during the process of 

hepatic fibrosis. In this study, we assessed protein fingerprint 

markers of the ECM in serum of patients with HCC compared 

to cirrhosis, chronic liver disease and healthy controls. We 

determined the diagnostic performance of these markers for 

HCC in order to gain insight into the mechanisms linking matrix 

turnover to HCC diagnosis. Methods Plasma from 86 HCC 

patients, age and sex-matched to 86 cirrhotics, 86 hepatitis B 

(HBV) non-cirrhotics and 86 healthy controls, were collected in 

a cross-sectional study. ECM markers of matrix metaloproteinases 

(MMP)-2, 7 or -9, derived collagen turnover of type I, III, 

IV, VI (C1M, C3M, C4M2, C6M) and elastin (ELM7), as well 

as formation of type III and IV collagen (Pro-C3, P4NP 7S) were 

measured by ELISA. Results Mean plasma Pro-C3 levels were 

significantly higher in HCC (33.12 ng/ml) than in cirrhotics 

(22.2 ng/ml), HBV non-cirrhotics (12.18 ng/ml) and healthy 

controls (9.03 ng/ml) (p


442 

Hepatocellular Carcinoma Surveillance Is Associated 

With Improved Survival And Can Be Targeted To High 

Risk Populations 

Thai Hong 1 , Paul Gow 2,10 , Michael A. Fink 3,11 , Anouk T. Dev 4 , 

Stuart K. Roberts 5 , Amanda J. Nicoll 6,7 , John Lubel 6,12 , Ian Kronborg 

8 , Niranjan J. Arachchi 8 , Marno C. Ryan 1,10 , William W. 

Kemp 5 , Virginia H. Knight 4 , Helen Farrugia 9 , Vicky Thursfield 9 , 

Paul V. Desmond 1,10 , Alex J. Thompson 1,10 , Sally Bell 1 ; 1 Gastroenterology, 

St Vincent’s Hospital, Melbourne, Melbourne, VIC, 

Australia; 2 Gastroenterology, Austin Hospital, Melbourne, VIC, 

Australia; 3 Surgery, Austin Hospital, Melbourne, VIC, Australia; 

4 Gastroenterology & Hepatology, Monash Medical Centre, Melbourne, 

VIC, Australia; 5 Gastroenterology & Hepatology, The 

Alfred Hospital, Melbourne, VIC, Australia; 6 Gastroenterology & 

Hepatology, Eastern Health, Melbourne, VIC, Australia; 7 Gastroenterology 

& Hepatology, The Royal Melbourne Hospital, Melbourne, 

VIC, Australia; 8 Gastroenterology & Hepatology, Western 

Hospital, Melbourne, VIC, Australia; 9 Victorian Cancer Registry, 

Cancer Council Victoria, Melbourne, VIC, Australia; 10 Medicine, 

The University of Melbourne, Melbourne, VIC, Australia; 11 Surgery, 

The University of Melbourne, Melbourne, VIC, Australia; 

12 Eastern Health Clinical School, Monash University, Melbourne, 

VIC, Australia 

Background: Hepatocellular carcinoma (HCC) incidence is heterogeneously 

distributed worldwide and is rising rapidly in 

many developed countries where the complete range of therapeutic 

options is available. Local epidemiology is important 

to accurately characterise the disease and allow strategies to 

effectively manage the problem. We performed the first population-based 

study in Australia to capture a clinical cohort and 

study the effect of aetiology, ethnicity and advanced therapeutic 

options on survival outcomes. Method: Incident cases of 

HCC (defined by AASLD diagnostic criteria or histology) were 

prospectively identified over a 12-month period (1 July, 2012 

to 30 June, 2013) from the population of Melbourne, Australia. 

Cases were captured from multiple sources including admissions 

and outpatient attendances to any of Melbourne’s seven 

tertiary hospitals, gastroenterology department, radiology, 

pathology and pharmacy databases as well as those registered 

by the Victorian Cancer Registry. Patients were followed up 

for at least 24 months with the primary endpoint being overall 

survival. Results: There were 272 incident cases identified. The 

major risk factors for liver disease were hepatitis C virus (HCV) 

infection (41%), alcohol (39%), and hepatitis B virus (HBV) 

infection (22%). Patients born overseas had up to 20 times the 

incidence rate of Australian-born patients. Regional variations 

of incidence within the study area correlated with percentage 

of overseas-born patients and HBV seroprevalence in those 

regions. Overall survival at 12 months was 57%. On multivariate 

analysis, the independent predictors of survival were age, 

the absence of cirrhosis, MELD score, AFP, tumour size, Barcelona 

Clinic Liver Clinic (BCLC) stage and being involved in an 

HCC surveillance program. Patients undergoing surveillance 

had significantly smaller tumour size (p


444 

Hepatic function and tumour burden are predictive 

of outcome in patients with hepatocellular carcinoma 

treated with transarterial chemoembolisation 

Martha Kirstein 1,2 , Nora Schweitzer 1,2 , Nazli Ay 1,2 , Christina 

Boeck 1,2 , Bernhard Meyer 1,3 , Thomas Rodt 1,3 , Wacker Frank 1,3 , 

Michael P. Manns 1,2 , Arndt Vogel 1,2 ; 1 Medical School Hannover, 

Hannover, Germany; 2 Gastroenterology, Hepatology, Endocrinology, 

Hannover, Germany; 3 Radiology, Hannover, Germany 

Background: Hepatocellular carcinoma (HCC) is one of the 

most lethal and prevalent cancers worldwide. Transarterial 

chemoembolization (TACE) is commonly used to act locally in 

the intermediate disease stage. Early randomized trials and 

more recent reviews and meta-analyses reported improved survival 

rates of patients with unresectable lesions managed with 

TACE so that TACE has been accepted as the standard treatment 

for intermediate stage disease. Methods: In this study, 

we characterized 487 HCC patients from Hannover Medical 

School, Germany, treated with TACE and evaluated the 

asscociation of their characteristics with survival. Results: 487 

HCC patients were treated at least once with TACE between 

2000 and 2013. Most patients (91.4%) were treated with conventional 

TACE using doxorubicin, ciplatin and/or mitomycin 

and lipiodol either alone or in combination with degradable 

starch microspheres (DSM, Spherex). Only minorities received 

transarterial embolisation (3.5%), transarterial chemoperfusion 

(3.1%) or TACE with drug-eluting beads (1.6%) as first transarterial 

treatment. 387 patients (79.3%) received TACE as first 

HCC-specific therapy. The mean number of received TACEs 

for all patients was 2.24±1.95(1-17) with a mean interval of 

140±257(33-2376) days between the first and second TACE. 

19.7%, 11.3% and 9% of the patients were diagnosed with 

one, two and three HCC nodules, respectively, whereas the 

remaining patients were diagnosed with more than three nodules. 

The mean diameter of the largest lesion was 59±36mm 

(4-226). 28.9% of the patients had at least one lesion ≥70mm. 

Hepatic function was well preserved in most patients with a 

mean Child-Pugh-Score A (6 points) and ALBI grade 2. After 

repeated TACEs there was no significant decrease of liver 

function observed as assessed by both classification systems. 

However, values of cholinesterase (CHE) were significantly 

decreased indicating CHE as a sensitive marker for patients 

treated with TACE (p


Overall survival after the diagnosis of HCC according to the HCV 

genotype. 

Disclosures: 

Raoel Maan - Consulting: AbbVie 


Gilead 




Heiner Wedemeyer - Advisory Committees or Review Panels: Transgene, MSD, 

Roche, Gilead, Abbott, BMS, Falk, Abbvie, Novartis, GSK, Roche Diagnostics; 

Grant/Research Support: MSD, Novartis, Gilead, Roche, Abbott, Roche Diagnostics; 

Speaking and Teaching: BMS, MSD, Novartis, ITF, Abbvie, Gilead 



Andres Duarte-Rojo - Advisory Committees or Review Panels: Gilead Sciences; 

Grant/Research Support: Vital Therapies; Speaking and Teaching: Roche 





Novartis 








Bart J. Veldt - Board Membership: GSK, Janssen Therapeutics 

Robert J. de Knegt - Advisory Committees or Review Panels: Roche, Norgine, 

Janssen Cilag, AbbVie; Grant/Research Support: Roche, Janssen Cilag, BMS, 

AbbVie; Speaking and Teaching: Gilead, Roche, Janssen Cilag, AbbVie 

The following authors have nothing to disclose: Frank Lammert, Bettina E. Hansen 

446 

Validation of the Metroticket calculator in liver transplant 

recipients that had a prior chemoembolization for 


Hyung-Don Kim 1 , Ju Hyun Shim 2 , Seungbong Han 3 , Mi-Jung Jun 2 , 

Yeonjung Ha 2 , Jihyun An 2 , Danbi Lee 2 , Kang Mo Kim 2 , Young- 

Suk Lim 2 , Han Chu Lee 2 , Young-Hwa Chung 2 , Yung Sang Lee 2 , 

Dong Jin Suh 4 ; 1 Department of Internal Medicine, Asan Medical 

Center, University of Ulsan College of Medicine, Seoul, Korea 

(the Republic of); 2 Department of Gastroenterology, Asan Medical 

Center, University of Ulsan College of Medicine, Seoul, Korea (the 

Republic of); 3 Department of Applied statistics, Gachon University, 

Gyeonggi-do, Korea (the Republic of); 4 Department of Internal 

Medicine, Vievis Namuh Hospital, Seoul, Korea (the Republic of) 

Background & Aims: The Metroticket calculator provides a continuum 

of survival probabilities for transplanted hepatocellular 

carcinoma (HCC) patients on the basis of tumor number, largest 

tumor diameter, and presence of microvascular invasion 

(MVI). This method had not yet been validated however in HCC 

patients with a previous history of transarterial chemoembolization 

(TACE). We performed this validation in our current study 

in liver transplant (LT) recipients. Methods: We enrolled 142 

patients with arterial enhancing HCC(s) on dynamic images 

treated with TACE between 1997 and 2011 at Asan Medical 

Center who were subsequent LT recipients. Tumor parameters 

measured by the enhancement radiological method pre-LT 

or by explant pathology post-LT were incorporated into the 

Metroticket calculator. The calculator was validated in terms 

of calibration and discrimination capacities for the entire study 

population and for subgroups undergoing living donor-related 

LT and infected with hepatitis B virus. Results: Study subjects 

received a median of 2 TACE sessions (range, 1-18) prior to 

LT, and most of these patients received a living donor graft 

(97.2%) and had HBV infection (89.4%). Through images and 

explants, 121 (85.2%) and 102 (71.8%) of our study subjects 

met the Milan criteria, respectively. Seventeen patients 

(12%) had no pathologic viable nodule, and 15 (10.6%) cases 

showed microvascular invasion. The mean 3-year and 5-year 

survival rates predicted by the radiological model for all 142 

patients were 76.4% and 70.1%, respectively, and fell perfectly 

within the 95% confidence intervals (CI) of the observed 

estimates (72.8-86.2% and 68.6-83.2%, respectively). In the 

pathological model incorporating microvascular invasion, the 

anticipated 5 year survival rate of 120 patients with viable 

nodules on explants was 69.5%, which also was inside the 

95% CI of the actuarial rates (67.9-83.5%). The c-index as a 

measure of discriminatory power was 0.61 and 0.62, respectively, 

for the 3- and 5-year predictions in the radiological 

model, and 0.72 for the 5-year prediction in the pathological 

model. All corresponding findings were comparable for 

subgroups of living donor-LT recipients and hepatitis B virus 

infected patients. Conclusion: A Metroticket calculation based 

on explant data can accurately predict post-LT survival in HCC 

patients with prior TACE. Estimate-based predictions before LT 

using imaging may have poorer discriminatory power compared 

to calibration. 

Disclosures: 

Young-Suk Lim - Advisory Committees or Review Panels: Bayer Healthcare, Gilead 

Sciences; Grant/Research Support: Bayer Healthcare, BMS, Gilead Sciences, 

Novartis 

Han Chu Lee - Grant/Research Support: Medigen Biotechnology Co., Novartis, 

Roche, Bayer HealthCare, Bristol-Myers Squibb, INC research, Boehringer Ingelheim, 

Taiho Pharmaceutical Co., Yuhan Co. 

The following authors have nothing to disclose: Hyung-Don Kim, Ju Hyun Shim, 

Seungbong Han, Mi-Jung Jun, Yeonjung Ha, Jihyun An, Danbi Lee, Kang Mo 

Kim, Young-Hwa Chung, Yung Sang Lee, Dong Jin Suh 

447 

Retreatment with TACE: Transition of Model to Estimate 

Survival in Ambulatory Hepatocellular carcinoma 

patients helps decision 

Young Youn Cho 1 , Su Jong Yu 1 , June Sung Lee 2 , Jeong-Ju Yoo 1 , 

Minjong Lee 1 , Donghyeon Lee 1 , Yuri Cho 1 , Eun Ju Cho 1 , Jeong- 

Hoon Lee 1 , Yoon Jun Kim 1 , Jung-Hwan Yoon 1 ; 1 Department of 

Internal Medicine and Liver Research Institute, Seoul National 

University College of Medicine, Seoul, Korea (the Republic of); 

2 Department of Internal Medicine, Ilsan Paik Hospital, Ilsan, Korea 


Background/Aims: Transarterial chemoembolization (TACE) is 

a major therapeutic modality for patients with unresectable 

hepatocellular carcinoma (HCC), but prognosis is variable. 

Model to Estimate Survival in Ambulatory Hepatocellular carcinoma 

patients (MESIAH) was recently developed and validated 

as a survival model to predict prognosis of patients with 

HCC. We aimed to develop a novel index using MESIAH for 

predicting who can benefit from repeated TACE. Methods:


From 2005 to 2008, 783 patients initially treated with TACE 

in Seoul National University Hospital (Seoul, Korea) were 

included in this study. Among them, 356 patients received 

second TACE session (TACE-2) and 196 patients received 

third TACE session (TACE-3) for recurred HCC. We assessed 

MESIAH score at baseline and prior to each TACE session, and 

designated patients into MESIAH transition group if MESIAH 

score increased before each TACE session. Cox regression and 

survival analysis were performed. Results: The mean age was 

56.3 ± 10.3 and hepatitis B virus were present in 76.0%. There 

were 43 (12.1%) patients in the pre-TACE-2 MESIAH transition 

group. There were no significant differences of Model for Endstage 

Liver Disease score (8.27 vs 8.89, P=0.28) and baseline 

MESIAH score (5.14 vs 5.02, P=0.41) between MESIAH transition 

and non-transition groups, respectively. MESIAH transition 

group showed shorter overall survival than the MESIAH 

non-transition group (6.0 versus 16.0 months, respectively; 

hazard ratio, 1.73; 95% confidence interval, 1.20-2.41). 

Similar results were observed when the definition of MESIAH 

transition was applied before TACE-3. Conclusions: MESIAH 

transition is a simple method which can be implicated in practice. 

MESIAH transition identifies patients who are unsuitable 

for retreatment with TACE and external validation is warranted. 

Disclosures: 

Yoon Jun Kim - Grant/Research Support: Bristol-Myers Squibb, Roche, JW Creagene, 

Bukwang Pharmaceuticals, Handok Pharmaceuticals, Hanmi Pharmaceuticals, 

Yuhan Pharmaceuticals; Speaking and Teaching: Bayer HealthCare 

Pharmaceuticals, Gilead Science, MSD Korea, Yuhan Pharmaceuticals, Samil 

Pharmaceuticals, CJ Pharmaceuticals, Bukwang Pharmaceuticals, Handok Pharmaceuticals 

The following authors have nothing to disclose: Young Youn Cho, Su Jong Yu, 

June Sung Lee, Jeong-Ju Yoo, Minjong Lee, Donghyeon Lee, Yuri Cho, Eun Ju 

Cho, Jeong-Hoon Lee, Jung-Hwan Yoon 

448 

Survival of Patients with Advanced Hepatocellular Carcinoma 

on Sorafenib: Retrospective Analysis from a 

Single Asian Center 

Yeonjung Ha, Danbi Lee, Ju Hyun Shim, Young-Suk Lim, Han Chu 

Lee, Young-Hwa Chung, Yung Sang Lee, Kang Mo Kim; Asan 

Medical Center, Seoul, Korea (the Republic of) 

Background: Sorafenib is the current standard of care for 

patients with advanced hepatocellular carcinoma (HCC); 

however actual treatment pattern varies from region to region 

as well as among physicians. Therefore, we retrospectively 

investigated the practice pattern and compared the effect of 

sorafenib on survival and progression with other treatment 

strategies. Methods: We conducted a single center retrospective 

study involving patients of Barcelona Clinic Liver Cancer 

C stage HCC on sorafenib with or without other treatments. 

Baseline characteristics, pre and intratreatment variables, and 

features regarding sorafenib administration were evaluated. 

The primary outcome measure was the overall survival (OS), 

and time-to-progression (TTP) was also calculated according 

to each treatment strategy. Results: A total of 658 patients 

(mean age 54.5 years, 83.3% male) were analyzed, with 293 

initially treated with sorafenib, 236 treated with transarterial 

chemoembolization (TACE) at first then switched to sorafenib, 

and 129 who received a combination therapy of TACE and 

sorafenib. The baseline characteristics of patients at the time 

of initial sorafenib administration were comparable across the 

three groups, Tumor-Node-Metastasis stage was IV in 96.3% 

and liver function was Child-Pugh A in 77.1%. The indication 

for sorafenib administration was mostly distant metastasis 

in all groups, 88.5% in the initial treatment group, 89.7% 

in the switched group and 80.6% in the combination group 

(p=0.43). The average dosage of sorafenib was similar across 

the groups (mean, 661.6 mg, p=0.18), whereas the treatment 

duration was significantly shorter in the switched group. 

Patients were withdrawn from sorafenib mostly because of disease 

progression, while 12.8% stopped the medication due 

to adverse reactions, most commonly hand-foot skin reaction 

(43.0%). The median OS was 11.8 months in sorafenib alone 

as compared with 13.5 months in the switched group and 

16.2 months in the combination group (p=0.13). In a subgroup 

analysis of patients with portal vein invasion but not with distant 

metastasis, combination therapy was associated with better OS 

(p=0.001) and longer TTP (p=0.020). In a multivariate model, 

only combined treatment strategy was significantly associated 

with longer OS (odds ratio [OR], 0.33, p=0.007) and TTP (OR, 

0.40; p=0.014). Conclusion: Neither switched therapy (TACE 

to sorafenib) nor combined approach was associated with a 

longer OS than sorafenib alone in patients with BCLC C HCC. 

However, in a subset of patients with portal vein invasion but 

not with distant metastasis, combined treatment with TACE and 

sorafenib showed better survival. 

Disclosures: 



Novartis 

Han Chu Lee - Grant/Research Support: Medigen Biotechnology Co., Novartis, 

Roche, Bayer HealthCare, Bristol-Myers Squibb, INC research, Boehringer Ingelheim, 

Taiho Pharmaceutical Co., Yuhan Co. 

The following authors have nothing to disclose: Yeonjung Ha, Danbi Lee, Ju Hyun 

Shim, Young-Hwa Chung, Yung Sang Lee, Kang Mo Kim 

449 

In patients with hepatocellular carcinoma (HCC) and 

macrovascular invasion (MVI) amenable to surgical 

resection, a survival similar to that observed on 

sorafenib can be achieved 

Charlotte E. Costentin 1 , Eric Vibert 2 , Françoise Roudot-Thoraval 3 , 

Thomas Decaens 4 , Nathalie Ganne-Carrié 5 , Bernard Paule 2 , 

Christian Letoublon 6 , Mélanie Chiaradia 7 , Julien Calderaro 8 , 

Rene Adam 2 , Ivan Bricault 9 , Giuliana Amaddeo 1 , Daniel Cherqui 

2 , Olivier Seror 10 , Didier Samuel 2 , Ariane Mallat 1 , Christophe 

Duvoux 1 , Alexis Laurent 11 ; 1 Hepatology, Hôpital Henri Mondor, 

Creteil, France; 2 Hepatobiliary surgery, Paul Brousse Hospital, 

Villejuif, France; 3 Public Health, Henri Mondor Hospital, Creteil, 

France; 4 hepato-gastroenterology, CHU Grenoble, Grenoble, 

France; 5 Hepatology, Jean Verdier Hospital, Bondy, France; 6 Surgery, 

CHU Grenoble, Grenoble, France; 7 Radiology, Henri Mondor 

Hospital, Creteil, France; 8 Pathology, Henri Mondor Hospital, 

Creteil, France; 9 Radiology, CHU Grenoble, Grenoble, France; 

10 Radiology, Jean Verdier Hospital, Bondy, France; 11 Surgery, 

Henri Mondor Hospital, Creteil, France 

Introduction: A median survival of 8 months has been reported 

in the SHARP trial in HCC+MVI treated with Sorafenib. In 

contrast, surgical retrospective studies report overall survivals 

ranging from 12 to 20 month in this setting. The aim of this 

study was to describe characteristics and overall survival in 

HCC+MVI patients treated with surgery or sorafenib as the 

first treatment step. Methods: 142 patients with HCC+MVI, 

without extra-hepatic spread, treated either with surgical resection 

(group 1; n=75) or sorafenib (group 2; n=67) in 4 French 

centers as first-line treatment after diagnosis of MVI were retrospectively 

reviewed. Results: Compared to patients in group 

2, patients in group 1 were younger (58.3±11.7 vs 65.5±9.6 

years, p


7.5% of the patients (n=5) in group 2 died within 3 months 

post-sorafenib initiation (p=0.196). In group 1, 29 patients 

(45%) had additional therapy either adjuvant or for recurrence 

(including TACE, chemotherapy (CT), sorafenib, thermo-ablation, 

additional resection) and one was transplanted. In group 

2, 20 (30%) patients had additional therapy either in addition 

to sorafenib or as second line (including TACE, CT, radiotherapy 

or radioembolization). Median overall survival (OS) was 

12.0 months (IC95%:5.5-18.5) in group 1 and 12.9 months 

(IC95%: 8.3-17.5) in group 2 (p=0.61). Taking into account 

the imbalance in baseline characteristics, a propensity analysis 

was performed, identifying 57 patients with similar characteristics 

with a median OS of 7.2 months (IC95%: 3.7-10.6) in the 

“resection” group and 12.6 months (IC95%: 7.1-18.0) in the 

“sorafenib” group (p=0.89). In multivariate analysis, prognostic 

factors associated with survival in the whole population (resection 

+ sorafenib) were PT time 50 mm HR=1.69, p=0.046), MVI involving 

1st order branch or main trunk (HR=1.948, p=0.042) 

and additionnal therapy (HR=0.500, p=0.024). Median OS 

in the resection+additional therapy group of patients (n=29) 

was 25.2 [8.8-41.6] months and 24.3 [15.9-32.7] months 

in the sorafenib+additional therapy group (n=20), (p=0.82). 

Conclusion: In patients with HCC+ MVI amenable to surgical 

resection, a survival similar to that observed on standard of 

care, sorafenib, can be achieved. Palliative surgery may be 

a valuable alternative in this setting. A multimodal treatment 

strategy was associated with prolonged survival. 

Disclosures: 



Roche 

Nathalie Ganne-Carrié - Advisory Committees or Review Panels: Roche; Speaking 

and Teaching: BMS, Gilead, Bayer 

Olivier Seror - Board Membership: Bayer Healthcare; Consulting: Olympus; 

Speaking and Teaching: Angiodynamics 






The following authors have nothing to disclose: Charlotte E. Costentin, Eric Vibert, 

Thomas Decaens, Bernard Paule, Christian Letoublon, Mélanie Chiaradia, 

Julien Calderaro, Rene Adam, Ivan Bricault, Giuliana Amaddeo, Daniel Cherqui, 

Ariane Mallat, Alexis Laurent 

450 

Prediction of liver decompensation and survival after 

TACE for hepatocellular carcinoma. Is there a role for 

non-invasive markers of fibrosis? 

Jean-Baptiste Hiriart 1 , François Franques 1 , Julien Vergniol 1 , Christophe 

Cassinotto 2 , Faiza Chermak 1 , Bruno Lapuyade 2 , Juliette 

Foucher 1 , Wassil Merrouche 1 , Jean-Frédéric Blanc 3 , Victor de 

Ledinghen 1 ; 1 Hepatology, Hôpital Haut-Lévêque, Pessac, France; 

2 Diagnostic and Interventional Imaging, Hôpital Haut-Lévêque, Pessac, 

France; 3 Hepatology, Hôpital Saint-André, Bordeaux, France 

Introduction: Non-invasive markers of liver fibrosis are useful to 

predict survival and liver-related events in patients with chronic 

liver disease. The aim of this study was to investigate the performance 

of these markers to predict hepatic decompensation and 

death after transarterial chemoembolization (TACE) for hepatocellular 

carcinoma (HCC). Patients and methods: From 2008 

to 2012, all consecutive patients undergoing a first TACE for 

HCC with a FibroScan and serum biomarkers (Fibrotest, APRI, 

Hepascore, FIB-4, AST/ALT ratio, NAFLD score) performed at 

baseline were included. Liver decompensation was defined as 

the occurrence of at least one of the following events within 6 

weeks after TACE: ascites, increase of bilirubin levels above 50 

mmol/L, acute kidney injury. Child-Pugh score and MELD score 

were calculated the day before TACE and one month after. 

Incident cases of death were prospectively collected. Results: 

103 patients were included (mean age 66 years, 85% men). 

In univariate analysis, factors associated with decompensation 

were albumin level 200 ng/ml (OR=2.51, CI95% 1.24-5.08, p=0.01) 

and albumin


for 70.9% and 12.6% of the total HCC cases (1292 patients) 

respectively. Out of the total cohort enrolled from 1980 to 

2005, the ratio of cryptogenic to CHB patients was 1:6.7; 

while from 2006 to 2015, the ratio of cryptogenic to CHB 

patients has increased significantly to 1:3.9. Both groups were 

comparable with regards to gender, Chinese ethnicity, smoking 

status, Child’s score, stage of HCC, portal vein invasion, 

AFP and platelet count. Relative to CHB group, cryptogenic 

HCC patients were older (68 versus 60 years old; p


The following authors have nothing to disclose: Atsuo Takigawa, Ryotaro Sakamori, 

Tomohide Tatsumi, Takayuki Yakushijin, Tadashi Kegasawa, Yuki Makino, 

Seiichi Tawara, Yoshinobu Saito, Yoshiki Onishi, Satoshi Tanaka, Kunimaro 

Furuta, Minoru Shigekawa, Naoki Hiramatsu 

Disclosures: 

The following authors have nothing to disclose: Li Huang, Kunsong Zhang, Wei 

Chen, Mengjun Hou, Yuyan Han, Fanyin Meng, Sharon DeMorrow, Xiaoyu Yin, 

Jiaming Lai, Bingyan Tan, Lijian Liang 

454 

Dysregulated peripheral and local endocannabinoid 

system promote tumor pathogenesis in biliary tract cancers 

Li Huang 1 , Kunsong Zhang 1 , Wei Chen 1 , Mengjun Hou 2 , Yuyan 

Han 3 , Fanyin Meng 4 , Sharon DeMorrow 5 , Xiaoyu Yin 1 , Jiaming 

Lai 1 , Bingyan Tan 2 , Lijian Liang 1 ; 1 Department of Pancreatobiliary 

Surgery, The First Affiliated Hospital, Sun Yat-sen University, 

Guangzhou, China; 2 Department of Nutrition, School of Public 

Health, Sun Yat-Sen University, Guangzhou, China; 3 Department 

of Medicine, Baylor and Scott & White Digestive Disease Research 

Center, Texas A&M HSC College of Medicine and Scott & White 

Hospital, Temple, TX; 4 Department of Medicine; Department of 

Research, Scott & White Digestive Disease Research Center; Texas 

A&M Health Science Center College of Medicine; Central Texas 

Veterans Health Care System, Temple, TX; 5 Department of Internal 

Medicine, Texas A&M Health Science Center College of Medicine; 

Baylor Scott & White Health Digestive Disease Research Center; 

Central Texas Veterans Health Care System, Temple, TX 

Background: The endocannabinoid system (ECS) is dysregulated 

in various liver diseases. Biliary tract cancers (BTCs) 

encompass gallbladder carcinoma, intrahepatic, perihilar and 

distal cholangiocarcinoma. Previously we have shown that the 

two major endocannabinoids (ECs), anandamide (AEA) and 

2-arachidonoyl glycerol (2-AG) exert opposing effects on proliferation 

of BTCs cell lines. However, regulation and clinical 

significance of ECS in BTCs remain inconclusive. Method: Levels 

of AEA and 2-AG in preoperative peripheral plasma and 

bile in 22 BTCs patients and 8 patients with benign biliary 

diseases, and paired resection samples in 15 BTCs patients 

were quantitated by gas chromatography/electron ionization 

(EI)-mass spectrometry (GC/MS). Expression and activity of 

the enzymes involved in ECs biosynthesis (phospholipase D 

for AEA and diacylglycerol lipases (DAGLs, including DAGLα 

and DAGLβ) for 2-AG) and ECs degradation (fatty acid amide 

hydrolase-1 (FAAH-1) for AEA and monoaclyglycerol lipase 

(MAGL) for 2-AG) were also analyzed. Levels of ECs were correlated 

with patients’ clinicopathologic features. Results: Level 

of AEA was lower in plasma and bile (median [interquartile 

range]: 1.804 pmol/mL [0.502 to 5.004 pmol/mL] vs 10.150 

pmol/mL [2.684 to 17.491 pmol/mL]) in BTCs than in benign 

biliary diseases, though the former didn’t reach significance. 

AEA was significantly lower in BTCs cancer nest than in adjacent 

normal tissue (22.01 fmol/mg [16.55 to 53.61 fmol/mg] 

vs 58.68 fmol/mg [25.36 to 68.97 fmol/mg]). Level of 2-AG 

was upregulated in plasma (180.0 pmol/mL [140.7 to 283.8 

pmol/mL] vs 42.3 pmol/mL [25.6 to 148.9 pmol/mL]) and 

bile in BTCs, though the latter didn’t reach significance. 2-AG 

was significantly higher in BTCs cancer nest than in adjacent 

normal tissue (1.967 pmol/mg [1.439 to 5.433 pmol/mg] vs 

1.101 pmol/mg [0.358 to 1.473 pmol/mg]). Upregulated 

expressions and activities of DAGLs, FAAH-1 and MAGL were 

demonstrated in BTCs. Lower cancer nest level of AEA, higher 

plasma and cancer nest level of 2-AG were correlated with 

more advanced tumor stage in BTCs. Conclusion: ECS is dysregulated 

in BTCs and it may foster a tumor promoting microenvironment. 

Modulation of ECS could be regards as a promising 

therapeutic strategy for BTCs. 

455 

Prospective cohort study for evaluating clinical effects 

and safety of intra-arterial infusion therapy of Cisplatin 

suspension in lipiodol combined with 5-fluorouracil and 

sorafenib for advanced hepatocellular carcinoma with 

macroscopic vascular invasion, without extra-hepatic 

spread 

Niizeki Takashi, Masahito Nakano, Takuji Torimura; Gastroenterology, 

Kurume University, school of medicine, Kurume, Japan 

Purpose Sorafenib is the standard first line treatment for patients 

with BCLC stage C hepatocellular carcinoma (HCC). However 

sorafenib monotherapy confers less than three months of 

actual survival benefit in both Western and Asian populations. 

Hepatic arterial infusion chemotherapy (HAIC) is recognized 

as a useful therapeutic option for advanced HCC in Japan, and 

response to HAIC is known as an important prognostic factor. 

While sorafenib has been proven to improve the prognosis in 

HCC patients with macroscopic vascular invasion (MVI), it is 

still unknown which is better, Sorafenib or HAIC. The aim of 

this multicenter none- randomized prospective cohort study was 

to investigate the efficacy and safety of HAIC compared with 

Sorafenib in patients with advanced HCC with MVI, without 

extra-hepatic spread (EHS) and Child-Pugh class A disease. 

Method: This study was conducted from April 2009 to March 

2014, total 64 HCC patients were registered. 44 were treated 

with HAIC, 20 were treated with Sorafenib. HAIC regimen 

comprised a combination of 50 mg fine powder formulation of 

Cisplatin in 5-10 ml lipiodol and contimuous infusion of 5-fluorouracil 

(1,500 mg/5 days). The primary efficacy endpoint 

was progression-free survival (PFS), while the secondary endpoints 

were Median survival time (MST), tumor response rate, 

and safety. PFS and MST were estimated by Kaplan- Meier 

method. Therapeutic effect was evaluated according to RECIST. 

Results: There were no statistica differences in clinical factors 

between two groups. PFS in HAIC and Sorafenib was 9.3 

months and 4.1 months respectively. (P = 0.002, 95% CI, 

0.221- 0.713) CR or PR rate in HAIC and Sorafenib was 71% 

and 10%.(P 0.001, 95% CI, 4.32- 106.09) MST in HAIC and 

Sorafenib was 30.0 months and 13.0 months respectively. (P 

= 0.016, 95% CI, 0.219- 0.854) Severe adverse events were 

observed in 5 patients. In Sorafenib group, 2 had hepatic 

failure, in HAIC group, 2 had hepatic failure, 1 had pseudo 

aneurysm. In HAIC group, treatment- related mortality was not 

observed. By multivariate analysis, independent predictor of 

survival were therapeutic effect (CR or PR, P= 0.009, 95% CI, 

0.220- 0.752), Child- Pugh score (score 5, P= 0.022, 95% 

CI, 0.191- 0.752), grade of portal vein invasion (trunk, P= 

0.002, 95% CI, 0.118- 0.614), and independent predictor of 

therapeutic effect was therapeutic regimen (HAIC, P 0.0001). 

Conclusion: In HAIC group, PFS was 9.3 months, MST was 

30.0 months and response rate was 74%, and these results 

were superior to Sorafenib. In conclusion, this regimen should 

be the first choice for patients with advanced HCC with MVI, 

without EHS and Child Pugh A disease. 

Disclosures: 

The following authors have nothing to disclose: Niizeki Takashi, Masahito 

Nakano, Takuji Torimura


456 

Impact of Hepatic p62 Overexpression on Survival of 

Patients with Hepatocellular Carcinoma 

Yasuji Komorizono 1 , Masaki Kitazono 2 , Sadao Tanaka 3 , Kazuhisa 

Nakashima 1 , Toshihiko Shibatou 1 , Katsumi Sako 1 ; 1 Hepatology, 

Nanpuh Hospital, Kagoshima, Japan; 2 Digestive Surgery, Nanpuh 

Hospital, Kagoshima, Japan; 3 Pathology, Nanpuh Hospital, 

Kagoshima, Japan 

Purpose: p62 is an adaptor or a scaffolding substrate between 

the ubiquitin–proteasome and the autophagy–lysosome pathways. 

p62 overexpression has been demonstrated to play a 

critical role in tumorigenesis of various malignancies, including 

hepatocellular carcinoma (HCC); however, the association 

between p62 overexpression and survival in HCC is currently 

unknown. Our purpose was to clarify whether overexpression 

of p62 impacts survival of patients with HCC. Methods: 

Between May 2003 and July 2013, we retrospectively identified 

78 patients with primary HCC (51 men and 27 women 

aged 34 to 81 years; mean, 68.4 years) who had undergone 

surgical resection and enrolled them in this study. We performed 

immunohistochemical analyses of their p62 expression 

and correlated the findings with annotated clinicopathologic 

data. Fifty-four patients (69%) were positive for hepatitis virus C 

antibody and 11 (14%) for hepatitis B antigen; the remaining 

13 patients (17%) were negative for both markers. We performed 

immunohistochemical analyses with antibody against 

p62 on surgically resected specimens and compared the overall 

survival of the p62-positive and -negative groups using the 

Kaplan–Meier method. We assessed differences between these 

two groups using the log-rank test. A Cox proportional hazards 

model was used to evaluate the interactions between the baseline 

characteristics and the impact of p62 overexpression on 

overall survival. Results:We detected p62 overexpression in 54 

patients (69%). Overall, 38 deaths (49%) occurred during follow-up 

(26 in the p62-positive and 12 in the -negative group). 

In the baseline characteristics, there was significant difference 

in the histological differentiation (poorly versus well and moderately) 

between the p62 positive group and the p62 negative 

group (p = 0.047). The overall survival rates at 5, 7, and 10 

years were 56%, 45%, and 23 % in the p62 positive group 

and 86%,58%, and 46% in the p62 negative group, respectively. 

The log-rank test revealed that the overall survival was 

significantly shorter in the p62 positive group than in the p62 

negative group (p = 0.046). Multivariate analyses showed that 

only p62 overexpression (HR, 2.29; 95% CI, 1.03–5.12; P = 

0.043) and main tumor size (HR, 1.17; 95% CI, 1.0–1.35; p 

= 0.044) were independently associated with overall survival. 

Conclusion: The results of the current study indicated that overexpression 

of p62 was closely associated with overall survival 

in patients with HCC. P62 could be used as a prognostic biomarker 

in patients with HCC. 

Disclosures: 

The following authors have nothing to disclose: Yasuji Komorizono, Masaki Kitazono, 

Sadao Tanaka, Kazuhisa Nakashima, Toshihiko Shibatou, Katsumi Sako 

457 

Improved Survival of Hepatocellular Carcinoma (HCC) 

in HIV/Hepatitis B Virus (HBV)-Coinfected Patients Who 

Are Diagnosed Through HCC Screening 

Maaz B. Badshah 1 , Meritxell Ventura-Cots 2 , Caitlin C. Citti 3 , Luciana 

Kikuchi 4 , Sonja Marcus 5 , Beatriz Minguez 6 , Ting-Yi Chen 7 , 

Maria D. Hernandez 8 , Judith Aberg 3 , Myron E. Schwartz 9 , Douglas 

Dieterich 3 , Norbert Bräu 5,3 ; 1 Indiana University School of 

Medicine, Indianapolis, IN; 2 Hospital Universitari Vall ‘Hebron, 

Barcelona, Spain; 3 Dept. of Medicine, Icahn School of Medicine 

at Mount Sinai, New York, NY; 4 Universidade de São Paulo, São 

Paulo-SP, Brazil; 5 James J. Peters VA Medical Center, Bronx, NY; 

6 Hospital Universitari Vall d’Hebron, Barcelona, Spain; 7 University 

of Texas Southwestern Medical Center, Dallas, TX; 8 University of 

Miami School of Medicine, Miami, FL; 9 Dept. of Surgery, Icahn 

School of Medicine at Mount Sinai, New York, NY 

BACKGROUND: Current recommendations for HCC screening 

in patients chronically infected with the hepatitis B virus 

(HBV) are based on a randomized controlled trial from China. 

However, no data are available on the effectiveness of HCC 

screening in HIV/HBV-coinfected patients. METHODS: HIV/ 

HBV-coinfected patients with HCC were retrospectively identified 

from 1992-2013 in 38 centers in 8 countries. Patients 

were considered screened if they presented with an abnormal 

alpha-fetoprotein level or imaging study, and not screened 

if they presented with symptoms. RESULTS: Among 74 HIV/ 

HBV-coinfected patients with HCC, 40 (60%) were screened. 

Compared to 34 unscreened patients, screened patients had 

similar mean age (49.4 vs. 50.7 years). However, screened 

patients had a lower mean Child-Pugh score (5.8 vs. 7.4, 

p=0.002), had a smaller median tumor size (3.6 vs. 8.7 cm, 

p=0.001), had a lower median alpha-fetoprotein level (107 

vs. 1,422 ng/ml, p=0.010), and more commonly met Milan 

criteria for liver transplantation (44% vs. 7%, p=0.001). They 

presented less frequently with portal vein thrombosis (23% vs. 

47%, p=0.031) and with advanced Barcelona-Clinic-Liver-Cancer 

stages C+D (36% vs. 78%, p=0.001), and more often 

received effective HCC therapy (80% vs. 27%, p< 0.001). With 

adjustment for lead time of 10.2 months, screened patients had 

a longer median survival (89 vs. 3.7 months, p=0.010, log 

rank). The actuarial 1-year survival rate was 70% in screened 

patients and 27% in unscreened patients. In multi-variable Cox 

proportional hazard analysis, screening was an independent 

predictor of survival (hazard ratio for death, 0.42; 95% confidence 

interval, 0.19 – 0.93; p=0.033), together with effective 

HCC therapy and Child-Pugh score. CONCLUSIONS: In 

HIV/HBV-coinfected patients with HCC, a diagnosis through 

screening was associated with earlier HCC stages, more HCC 

therapy, and independently predicted better survival.


5.34, p


respectively. All searches and data extraction were performed 

independently by 2 authors. Included studies were published in 

2000 or after and had ≥100 patients undergoing surveillance 

as defined above. Analysis was via random-effects models. 

Results: A total of 14 studies with 15,429 (12,828 CHB; 2,601 

cirrhosis) met inclusion criteria. Overall adherence was 61% 

(95% CI 56-67%). Cirrhosis patients had significantly higher 

adherence than CHB patients, 65% (95% CI 61-69%) vs. 58% 

(95% CI 49-67%), p


461 

Etiological difference in body composition of patients 

with hepatocellular carcinoma 

Ryosuke Tateishi 1 , Naoto Fujiwara 1 , Hayato Nakagawa 1 , Yotaro 

Kudo 1 , Ryo Nakagomi 1 , Mayuko Kondo 1 , Takuma Nakatsuka 1 , 

Tatsuya Minami 1 , Masaya Sato 1 , Koji Uchino 1 , Kenichiro Enooku 1 , 

Yuji Kondo 1 , Yoshinari Asaoka 1 , Yasuo Tanaka 1 , Kyoji Moriya 1 , 

Shuichiro Shiina 2 , Kazuhiko Koike 1 ; 1 Department of Gastroenterology, 

The University of Tokyo Hospital, Tokyo, Japan; 2 Department 

of Gastroenterology, Juntendo University, Tokyo, Japan 

Background: We have reported that sarcopenia, intramuscular 

fat deposition, and visceral adiposity indicate poor prognosis 

of patients with hepatocellular carcinoma (HCC). To clarify the 

etiological difference in body composition of HCC patients, 

we compared visceral adipose tissue index (VATI), subcutaneous 

adipose tissue index (SATI), and visceral to subcutaneous 

adipose tissue ratio (VSR) among etiological groups in the 

cohort. Methods: We enrolled naïve HCC patients diagnosed 

from January 2004 to December 2009 in our department. 

Patients were divided into 3 groups according to chronic viral 

infection: hepatitis C virus (HCV), hepatitis B virus (HBV) and 

non-viral etiology (NBNC). Those who were infected with both 

hepatitis viruses were excluded. To minimize the tumor effect 

on body composition, those with tumor larger than 5 cm were 

also excluded. We assumed that visceral fat accumulation 

increases inversely proportion to impact of virus related-risk of 

HCC development (i.e., HCV>HBV>NBNC). Each component 

of body composition was compared between two groups using 

HCV group as a control. Results: Baseline characteristics of 

enrolled patients were as follows: HCV group (N = 534, M/F 

= 311/223, median age = 71), HBV group (N = 84, M/F 

= 67/17, age = 61), and NBNC group (N = 126, M/F = 

97/29, age = 71). Median (interquartile range [IQR]) tumor 

size was 2.3 (1.8-2.9) cm. Number of tumor nodule was 1 in 

404, 2-3 in 248, and more than 3 in 92 patients. Child-Pugh 

class was A in 574, B in 164, C in 6. Significant difference 

was observed in VATI between HBV and HCV groups in males 

(P = 0.02) as well as between NBNC and HCV groups in 

males (P 0.05). 

11 patients were treated with CE + sorafenib. Advanced BCLC 

stage disease, cirrhosis, and increasing MELD were statistically 

significant predictors of decreased survival in univariate models. 

In multivariate models the only statistically significant predictors 

of improved survival included hepatitis B etiology and 

treatment with resection or ablation (p


463 

Progranulin Autoantibodies and Genetic Variation in 

Patients with Cholangiocarcinoma 

Vincent Zimmer 1 , Lorenz Thurner 2 , Monica Acalovschi 3 , Michael 

Pfreundschuh 2 , Frank Lammert 1 ; 1 Department for Medicine II, Saarland 

University Medical Center, Homburg, Germany; 2 Department 

of Medicine I, Saarland University Medical Center, Homburg, Germany; 

3 Department of Medicine III, University Iuliu Hatieganu, 

Cluj-Napoca, Romania 

Background: Recently, the novel growth factor and TNF signalling 

adaptor progranulin (PGRN) has been implicated in 

the inflammation-driven pathogenesis of cholangiocarcinoma 

(CCA).(1,2) Neutralizing PGRN autoantibodies have been 

described in autoimmune diseases, including inflammatory 

bowel disease (IBD).(3,4) Therefore, our aim now was to 

assess PGRN autoantibody incidence and potential effects of 

the common PGRN gene variant rs5848 (known to be associated 

with reduced PGRN serum concentrations) on susceptibility 

and PGRN autoantibody formation in a large European 

CCA population.(5) Patients & Methods: Overall, 221 individuals 

with CCA (males n = 131, age 66 ± 11 years) originating 

from Germany (n = 164) and Romania (n = 57) and 295 

CCA-free control individuals were genotyped. The common 

non-synonymous single nucleotide polymorphism (SNP) rs5846 

was genotyped by PCR-based assays with 5’-nuclease and 

fluorescence detection (TaqMan). PGRN autoantibodies were 

determined by ELISA.(4) Results: rs5848 genotype frequencies 

in the control group were consistent with the Hardy-Weinberg 

equilibrium (HWE), with a call rate > 99 % indicating robust 

genotyping. The association tests did not provide evidence 

for genetic CCA risk modulation by the PGRN variant (Odds 

ratio = 0.92; 95 % confidence interval = 0.71-1.19; P>0.05). 

Seven of 61 patients (11.5%) tested positive for PGRN autoantibodies, 

with no significant sex differences (3/32 males, 4/29 

females). Considering rs5848 variation in the subgroup with 

available PGRN autoantibody status, no significant differences 

in allele and genotype distribution were observed. Conclusions: 

Our preliminary data indicate PGRN autoantibodies as a novel 

potential tumor antigen in a defined subset of CCA patients. 

Extension of the current study in larger sample sizes and delineation 

of potential implications in terms of CCA subtypes and 

prognostic relevance of PGRN autoimmunity is warranted. References: 

1. Frampton et al. Gut 2012 2. Frampton et al. Am 

J Physiol Gastrointest Liver Physiol 2012 3. Thurner et al. J 

Autoimmun 2013 4. Thurner et al. Dig Dis Sci 2014 5. Hsiung 

et al. J Neurol Sci 2011 

Disclosures: 

The following authors have nothing to disclose: Vincent Zimmer, Lorenz Thurner, 

Monica Acalovschi, Michael Pfreundschuh, Frank Lammert 

464 

The effect of additional transarterial chemoembolization 

after combined chemoradiation therapy for locally 

advanced hepatocellular carcinoma with portal vein 

thrombosis 

Ja Kyung Kim, Jung Il Lee, Jun Won Kim, Ik Jae Lee, Seung-Moon 

Joo, Kwang-Hun Lee, Eun-Suk Cho, Tae Joo Jeon, Jae Keun Kim, 

Nomi Park, Ah-Young Kang, Kwan Sik Lee; Yonsei University College 

of Medicine, Seoul, Korea (the Republic of) 

Backgrounds: Combined chemoradiation therapy (CCRT) followed 

by intraarterial chemotherapy (IAC) has been reported 

to be beneficial for locally advanced hepatocellular carcinoma 

(HCC) with portal vein thrombosis (PVT). However, hepatic 

arterial chemoembolization (TACE) was generally not accepted 

for this situation. The aim of this study is to evaluate the role of 

adding TACE instead of IAC after CCRT. Methods: Thirty-seven 

patients were treated by CCRT as the initial treatment for HCC 

with PVT between 2009 and 2014. IAC through hepatic arterial 

chemoport and intermittent TACE (Group A) followed in 

18 patients and only IAC were repeated in 19 patients (Group 

B). We excluded the patients who had extra-hepatic metastasis, 

active peptic ulcer, and inadequate hepato-renal function. 

For CCRT, infusion of 5-FU (500 mg/day) via chemoport 

was given during the first and last five days of external radiation 

therapy (RTx) for 5 weeks. For IAC, infusion of cisplatin 

(70~100mg/m2 for one day) with 5-FU (750~1000mg for 

3 days) was repeated monthly. Results: Mean age was 55 

year; male to female was 32 to 5. Underlying liver diseases 

were hepatitis B, hepatitis C and non-B/C in 26, 2 and 10 

patients, respectively. Mean follow-up duration was 14 months 

(3-65 months). The objective response (CR+PR) rates in tumor 

size after 1 month after CCRT were 45.9%. The mean level 

of αFP after 1 month after CCRT were decreased in 81% of 

patients. During follow up, complete response was noted in 

3 patients. One patient underwent resection for cure. Median 

overall survival duration was 12.4 months. Overall survival of 

group A was significantly better than that of group B (mean 

42.5 vs. 12.2 months, p=0.001). Conclusion: Adding intermittent 

transarterial chemoembolization contributed survival gain 

following combined chemoradiation therapy for advanced 

hepatocellular carcinoma with portal vein thrombosis. Further 

prospective randomized study comparing TACE with TAC after 

CCRT is warranted. 

Disclosures: 

The following authors have nothing to disclose: Ja Kyung Kim, Jung Il Lee, Jun 

Won Kim, Ik Jae Lee, Seung-Moon Joo, Kwang-Hun Lee, Eun-Suk Cho, Tae Joo 

Jeon, Jae Keun Kim, Nomi Park, Ah-Young Kang, Kwan Sik Lee 

465 

Clinicopathologic analysis of patients with HBV-positive, 

HCV-positive and non-B non-C hepatocellular carcinomas 

Jun Akiba 1 , Osamu Nakashima 2 , Yoshiki Naito 1 , Hironori 

Kusano 1 , Reiichiro Kondo 1 , Hirohisa Yano 1 ; 1 Department of 

Pathology, Kurume University School of Medicine, Kurume, Japan; 

2 Department of Clinical Laboratory Medicine, Kurume University 

Hospital, Kurume, Japan 

[aim] The chronic infection of hepatitis B virus (HBV) and hepatitis 

C virus (HCV) is strongly associated with HCC. However, 

recent developments of anti-viral treatments for HBV and HCV 

contribute to reduce virus-associated HCC. On the other hand, 

HCCs, which are negative for both hepatitis B surface antigen 

(HBs-Ag) and anti-hepatitis C antibody (HCV-Ab), are increasing 

in number in developed nations. The aim of this study is 

to clarify the clinicopathologic features of surgically resected 

HCCs with different etiologies. [Materials and methods] Surgically 

resected 1430 cases (1598 nodules) at our hospital from 

1991 to 2013 were enrolled in this study. All cases did not 

have any previous treatments for HCC. HBV-related HCC (HBV- 

HCC) and HCV-related HCC (HCV-HCC) were defined as positive 

for serum HBs-Ag and serum HCV-Ab, respectively. Non 

HBV and non HCV-related HCC (NBNC-HCC) was defined 

as negative for both HBs-Ag and HCV-Ab. The cases which 

were positive for both HBs-Ag and HCV-Ab were excluded. 

Clinicopathologic features were compared. [Results] Out of 

1430, 253, 949 and 228 cases were HBV-HCC, HCV-HCC 

and NBNC-HCC, respectively. Patient age of HBV-HCC (55.8) 

was significantly younger than those of HCV-HCC (67.6) and 

NBNC-HCC (68.5). Tumor size was the smallest in HCV-HCC


(31.5 mm), followed by HBV-HCC (40.0 mm) and NBNC-HCC 

(46.4 mm). Serum AFP and DCP were significantly higher in 

HBV-HCC than in HCV-HCC. Macroscopically, the percentage 

of HCC with indistinct margin was significantly higher 

in HCV-HCC (10.7%) compared with HBV-HCC (3.2%) and 

NBNC-HCC (3.6%). On the other hand, HBV-HCC (20.7%) significantly 

more frequently showed confluent multinodular type 

compared with HCV-HCC (11.2%) and NBNC-HCC (8.7%). 

The number of well-differentiated HCC and poorly differentiated 

HCC were significantly higher in HCV-HCC (11.1%) and 

in HBV-HCC (22.1%), respectively. Portal vein invasion was 

significantly more frequent in HBV-HCC (67.4%) compared 

with HCV-HCC (46.1%) and NBNC-HCC (56.5%). Patients 

with NBNC-HCC significantly had higher rate of a history of 

diabetes and alcohol intake. Out of 228 NBNC-HCC patients, 

21.3% showed steatohepatitis in the background liver. About 

half of them (11.2%) had a history of alcohol intake. The 

residual cases (10.1%) were defined as non-alcoholic steatohepatitis. 

Also, 19.7% of NBNC-HCC showed positivity of at 

least one of serum HBV-related markers except HBs-Ag. [conclusions] 

HBV-HCC, HCV-HCC and NBNC-HCC had different 

clinicopathologic features. HBV-HCC showed aggressive biologic 

features. NBNC-HCC was closely associated with not 

only diabetes and alcohol intake but also HBV infection. 

Disclosures: 

The following authors have nothing to disclose: Jun Akiba, Osamu Nakashima, 

Yoshiki Naito, Hironori Kusano, Reiichiro Kondo, Hirohisa Yano 

466 

The presence of histological abnormalities even after the 

elimination of hepatitis C virus 

Nobuhiro Aizawa 1 , Kyohei Kishino 1 , Yoshihiro Shimono 1 , Chikage 

Nakano 1 , Kunihiro Hasegawa 1 , Ryo Takata 1 , Kazunori 

You 1 , Akio Ishii 1 , Tomoyuki Takashima 1 , Yoshiyuki Sakai 1 , Naoto 

Ikeda 1 , Takashi Nishimura 1 , Hiroki Nishikawa 1 , Yoshinori Iwata 1 , 

Hirayuki Enomoto 1 , Hiroko Iijima 1 , Seikan Hai 2 , Jiro Fujimoto 2 , 

Shuhei Nishiguchi 1 ; 1 Division of Hepatobiliary and Pancreatic Diseases, 

Department of Internal Medicine, Hyogo collage of medicine, 

Nishinomiya, Japan; 2 Division of surgery, Hyogo college of 

medicine, Nishinomiya, Japan 

Background: Despite the improved histological findings in the 

liver after hepatitis C virus (HCV) eradication, hepatocellular 

carcinoma (HCC) occurs in some patients with a sustained 

virological response (SVR) to interferon therapy. However, the 

mechanism of carcinogenesis in virus-eliminated liver tissues 

remains unclear. The aim of this study is to clarify the characteristics 

of SVR-related hepatocarcinogenesis. Subjects and 

Methods: A total of 43 patients with SVR were enrolled. Fifteen 

patients had HCC (Group A), and the remaining 28 patients 

did not (Group B). We pathologically examined the non-cancerous 

hepatic tissues of Group A patients, and compared those 

tissues with the liver tissues of Group B patients by electron 

microscopy (EM). We evaluated the degree of the dilatation of 

vesicular endoplasmic reticulum on a scale of 0 to 3. We also 

evaluated the mitochondrial alterations on a scale of 0 to 10 

according to the following points: (1) Dense granules, (2) Paracrystalline 

inclusions, (3) Vacuole formation, (4) Irregularity 

of mitochondrial shapes, and (5) Lack of mitochondrial membrane. 

This study was conducted in accordance with the Helsinki 

declaration and written informed consents were obtained 

from all patients before the study. Results: Regarding dilatation 

of vesicular endoplasmic reticulum, elevated scores (> 2 points) 

were found in 14.3% (4/28) of the Group B patients and 

in 92.9% (13/15) of the Group A patients. Regarding the 

morphological mitochondrial alterations, elevated scores (> 

4 points) were found in 3.6% (1/28) of the Group B patients 

and in 80.0% (12/15) of the Group A patients. Scores of 

the alterations regarding the mitchondria and the vesicular 

endoplasmic reticulum were higher in the HCC Group than in 

the non-HCC Group. The morphological abnormalities were 

not associated with the histological findings, including activity 

grade and fibrosis stage. Conclusion: The EM findings 

revealed the presence of histological abnormalities even after 

the elimination of HCV. In particular, remarkable morphological 

abnormalities were observed in non-cancerous hepatic 

tissues of HCC patients. Their histological alterations after SVR 

should be correlated with hepatocarcinogenesis. 

Disclosures: 

The following authors have nothing to disclose: Nobuhiro Aizawa, Kyohei 

Kishino, Yoshihiro Shimono, Chikage Nakano, Kunihiro Hasegawa, Ryo Takata, 

Kazunori You, Akio Ishii, Tomoyuki Takashima, Yoshiyuki Sakai, Naoto Ikeda, 

Takashi Nishimura, Hiroki Nishikawa, Yoshinori Iwata, Hirayuki Enomoto, 

Hiroko Iijima, Seikan Hai, Jiro Fujimoto, Shuhei Nishiguchi 

467 

Prognostic Score predicting overall survival of Patients 

with Intermediate Stage of Hepatocellular Carcinoma 

after Transarterial Chemoembolization 

Saharat Jarupongprapa, Supot Nimanong, Siwaporn Chainuvati, 

Phunchai Charatcharoenwitthaya, Tawesak Tanwandee, Watcharasak 

Chotiyaputta; Gastroenterology, Internal Medicine, Siriraj 

Hospital, Mahidol University, Bangkok, Thailand 

Background: Transarterial chemoembolization (TACE) is the 

standard of care for patients (pts) with intermediate stage 

of hepatocellular carcinoma (HCC). Multiple TACE sessions 

are common in treatment of HCC which can result in shorten 

survival. Several baseline characteristics and dynamic tumor 

response after the first TACE were studied to select the patients 

who are not beneficial form repeated TACE. Objectives: To 

determine factors associated with overall survival (OS) after first 

TACE and to develop a prognostic score to identify candidates 

who have benefit form repeated TACE. Methods: A retrospective 

study between 2007 and 2012, all HCC pts underwent 

at least two consecutive TACE within 90 days. Pts with Child- 

Pugh (CTP) C, Barcelona Clinic Liver Cancer (BCLC) stage C 

and ruptured HCC at presentation were excluded. Baseline 

characteristics, liver function and dynamic tumor response after 

first TACE were analyzed on the median OS. Results: 216 

pts were included in this study. 75% were male, mean age 

was 59.2 years, and 74.5% were viral associated HCC. At 

baseline, mean tumor size was 7.9 cm, 62.5% was unilobar, 

and 60.6% was multifocal HCC. The median OS was 17.6 

months (15.3-19.9). 89.8% of pts (n=194) died during study 

period. On multivariate analysis, five parameters were significant 

factors to predict OS including baseline CTP B [HR1.77, 

p=0.04], BCLC stage B [HR1.95, p=0.001], increased CTP 

score after TACE [+1 (HR2.29), +2 (HR11.74), p 200 IU/L with < 50% reduction after TACE 

[HR2.07, p=0.003], and radiologic response (stable or progressive 

disease on mRECIST) [HR1.56, p=0.01]. A prognostic 

score using five parameters was developed and divided into 

two groups (


and high major complications. Retreatment with TACE should 

be avoided. 

Figure 1. Overall survival analysis according to SMART 

Disclosures: 

Tawesak Tanwandee - Grant/Research Support: MSD, BMS, Fibrogen, Biotron, 

Celcion 

The following authors have nothing to disclose: Saharat Jarupongprapa, Supot 

Nimanong, Siwaporn Chainuvati, Phunchai Charatcharoenwitthaya, Watcharasak 

Chotiyaputta 

468 

Combination of modified Response Evaluation Criteria 

in Solid Tumors and hand-foot-skin reaction: a new 

prognostic evaluation for HCC patients treated with 

sorafenib and transarterial chemoembolization 

Wenjun Wang 1 , Yan Zhao 1 , Wei Bai 1 , Lei Liu 1 , Man Yang 1 , Hongwei 

Cai 3 , Lei Zhang 1 , Zhanxin Yin 1 , Zhuoli Zhang 1 , Daiming 

Fan 1 , Jielai Xia 2 , Guohong Han 1 ; 1 Xijing Hospital of Digestive 

Diseases, Xijing Hospital, Fourth Military Medical University, Xi’an, 

China; 2 Department of Medical Statistics, Fourth Military Medical 

University, Xi’an, China; 3 Information Center, School of Stomatology, 

Fourth Military Medical University, Xi’an, China 

The aim of this study was to combine the modified Response 

Evaluation Criteria in Solid Tumors (mRECIST) and hand-footskin 

reaction (HFSR) to establish a new prognostic evaluation 

for hepatocellular carcinoma (HCC) patients undergoing 

sorafenib and transarterial chemoembolization (TACE). 176 

consecutive intermediate-advanced HCC patients treated with 

combination therapy were enrolled. The therapeutic responses 

were assessed by mRECIST and HFSR criteria at 1, 2 and 3 

months, respectively. Uni/multivariate Cox regressions were 

used to investigate the earliest time when treatment responses 

could be accurately assessed. Then according to the mRECIST 

and HFSR assessed at that time, SMART (Sorafenib with Modified 

RECIST Assessment plus hand-foot-skin Reaction in TACE) 

prognostic evaluation was developed: SMART A, responders 

on both assessments; SMART B, responders on either of assessment 

and SMART C, non-responders on both assessments. 

Moreover, we explore the prognostic value of SMART for 

predicting overall survival(OS) in comparison with mRECIST 

and HFSR about likelihood ratio, Akaike information criterion 

(AIC) and C-index respectively. The earliest time at which the 

responses of mRECIST and HFSR correlated with the survival 

was 2 months after therapy. The SMART stratified patients with 

three different prognosis; the SMART A had the longest median 

OS, followed by SMART B and SMART C (30.5, 17.4, and 

8.3 months, respectively; P


6% and 1% patients, respectively. In explant analysis, tumor 

was uninodular in 45% and moderately differentiated in the 

majority of cases (66%). Median HCC size was 28mm. Vascular 

invasion and satellite nodules were observed in 24.5% 

and 25% of patients, respectively. In 818 patients that survived 

beyond the immediate post-transplant period, mean follow-up 

was 27,7 months (±23,8), an overall survival was 70% in 5 

years. Recurrence occurred in 8/818 (8%) cases, at a mean 

time of 15 months (1.5-76m). Sites of recurrence were 40% in 

liver, extrahepatic in 46% and both hepatic and extrahepatic 

in 14%. Vascular invasion and alpha-fetoprotein (AFP) level 

before liver transplantation were risk factors for tumor recurrence. 

The presence of HCC recurrence was directly related 

to poor survival. Female gender, need for re-transplantation, 

vascular invasion and explant outside the Milan criteria were 

also predictors of poor survival. Conclusion: Liver transplantation 

for hepatocellular carcinoma in Brazil was associated with 

an overall survival of 70% in 5 years. HCC recurrence ocurred 

in 8% of patients. The presence of vascular invasion and AFP 

before liver transplantation were associated with increased 

risk of tumor recurrence. Female, vascular invasion, explant 

outside the Milan criteria and HCC recurrence were related to 

poor survival. 

Disclosures: 

The following authors have nothing to disclose: Aline Chagas, Luciana Kikuchi, 

Guilherme Felga, Angelo A. Mattos, Renato F. Silva, Rita de Cássia M. da Silva, 

Fernanda Branco, Marcio D. Almeida, Ilka F. Boin, José H. Garcia, Luiz C. D’Albuquerque, 

Flair J. Carrilho 

470 

MESIAH score is an effective subclassification tool in 

Barcelona Clinic Liver Cancer stage B: comparison with 

other proposed methods 

Jeong-Ju Yoo 1 , Su Jong Yu 1 , Eun Ju Cho 1 , Jeong-Hoon Lee 1 , June 

Sung Lee 2 , Yoon Jun Kim 1 , Jung-Hwan Yoon 1 ; 1 Department of 

Internal Medicine and Liver Research Institute, Seoul National 


2 Department of Internal Medicine, Ilsan Paik Hospital, Inje University 

College of Medicine, Goyang, Korea (the Republic of) 

Background: The intermediate stage of hepatocellular carcinoma 

(HCC) is a highly heterogeneous population, therefore 

many models to predict survival of patients has been proposed. 

The aim of this study is to evaluate the prognostic performance 

in intermediate HCC among four models; Model to Estimate 

Survival in Ambulatory Patient (MESIAH), original Barcelona 

Clinic Liver Cancer (BCLC) B subclassification, modified model 

A and modified model B. Methods: From January 2005 to 

December 2006, a total of 1184 HCC patients who initially 

treated with TACE were enrolled. Among them, 235 (19.8%) 

patients were classified as BCLC stage B. Four subclassification 

systems were tested; MESIAH, original BCLC B subclassification 

(B1, B2, B3, B4), modified model A (B1, B2, B3+B4), and 

modified model B (B1, B2+B3, B4). The predictive accuracies 

of the four systems were compared based on c-statistic (c-index) 

together with its confidence interval (CI). Results: Patients in 

the cohort had a median age of 57 years and 80.9% were 

men. 81.7% had a Child–Pugh class A and Hepatitis B virus 

infection was present in 71.1% and the median overall survival 

was 57.8 months. The MESIAH score had a highest degree 

of discrimination, with a C-statistic of 0.617 [95% confidence 

interval (CI), 0.564–0.670, hazard ratio 2.06 (1.72-2.48), 

p


*all from bilirubin increase 

**AFP reduction from baseline of ≥20% AFP or ≥50% AFP in 

patients with baseline value ≥20ng/mL 

Disclosures: 

Robin K. Kelley - Advisory Committees or Review Panels: Acceleron Inc.; Consulting: 

Eli Lilly & Co., Acceleron Inc.; Grant/Research Support: Exelixis, Inc., 

Celgene Corp., Eli Lilly & Co., Regeneron Pharmaceuticals, Tekmira Pharmaceuticals, 

Novartis, Sanofi 




The following authors have nothing to disclose: Varun Saxena, Neil Mehta, K. 

Pallav Kolli, Robert Kerlan, Francis Y. Yao, Albert Chang 

two groups (P = 0.666). Multivariate analysis identified ICG 

R and solitary tumor as independent factors associated with 

disease-free survival. Conclusion: The two treatment modalities 

attained similar survival benefits in the management of initial 

recurrent HCC after curative treatment. In addition to host and 

tumor factors, time from primary HCC development to recurrence 

was a prognostic factor for recurrence of HCC. 

Disclosures: 

Kazuaki Chayama - Consulting: AbbVie; Grant/Research Support: Ajinomoto, 

Astellas, Asuka, Bayer, Daiichi Sankyo, Dainippon Sumitomo, Eisai, Janssen, 

Kowa, Mitsubishi Tanabe, MSD, Eli Lily, Nippon Kayaku, Nippon Shinyaku, 

Otsuka, Roche, Takeda, Toray, Torii, Tsumura, Zeria; Speaking and Teaching: 

Eisai, Ajinomoto, AbbVie, Abott, Astellas, AstraZeneca, Asuka, Bayer, BMS, 

Chugai, Daiichi Sankyo, Dainippon Sumitomo, J&J, Jimro, Miyarisan, MSD, 

Nihon Kayaku, Olympus 

The following authors have nothing to disclose: Takayuki Fukuhara, Hiroshi 

Aikata, Fumi Shinohara, Norihito Nakano, Yuki Nakamura, Masahiro Hatooka, 

Kei Morio, Tomoki Kobayashi, Yuuko Nagaoki, Tomokazu Kawaoka, Masataka 

Tsuge, Akira Hiramatsu, Michio Imamura, Yoshiiku Kawakami 

472 

Survival analysis of second hepatectomy versus radiofrequency 

ablation for initial recurrent small hepatocellular 

carcinoma after curative treatment 

Takayuki Fukuhara, Hiroshi Aikata, Fumi Shinohara, Norihito 

Nakano, Yuki Nakamura, Masahiro Hatooka, Kei Morio, Tomoki 

Kobayashi, Yuuko Nagaoki, Tomokazu Kawaoka, Masataka 

Tsuge, Akira Hiramatsu, Michio Imamura, Yoshiiku Kawakami, 

Kazuaki Chayama; Hiroshima university, Hiroshima, Japan 

Background and Aim: Japanese and American clinical practice 

guidelines recommend hepatectomy and radiofrequency 

ablation (RFA) as curative treatments for small hepatocellular 

carcinoma (HCC; ≤ 3 nodules and each ≤ 3 cm in diameter). 

However, the algorithms recommended in the above-mentioned 

guidelines are for primary HCC, not recurrent HCC. The treatment 

algorithm for recurrent HCC is not yet reach consensus. 

The aim of this retrospective study is to compare the efficacy 

of hepatectomy and RFA as the treatment of initial recurrence 

after curative treatment. Methods: The following inclusion criteria 

were used: 1) initial recurrent HCC after curative treatment, 

2) ≤ 3 nodules and each ≤ 3 cm in diameter, 3) no vascular 

invasion and no extrahepatic metastasis, and 4) Child-Pugh 

class A or B. From January 2001 to December 2014, one 

hundred and ninety two patients who met the inclusion criteria 

were evaluated. Patients were divided into 2 groups: second 

hepatectomy group (n = 120) and RFA group (n = 72). To overcome 

bias due to the different distribution of covariates for the 

two groups, a one-to-one match was created using propensity 

score analysis. After matching, we compared overall survival 

(OS), disease-free survival (DFS) between these groups. Results: 

1) After one-to-one matching, there was no significant difference 

in clinical backgrounds between second hepatectomy 

group (n = 62) and RFA group (n = 62). 2) The rate of OS at 

3, 5 and 10 years was 77.1%, 59.0% and 38.9% in the second 

hepatectomy group and 85.3%, 58.6% and 35.1% in the 

RFA group, respectively. There was no significant difference in 

OS between these two groups (P = 0.986). Multivariate analysis 

identified several host and tumor factors (age


Disclosures: 

Armando Carvalho - Advisory Committees or Review Panels: Gilead Sciences, 

Bristol-Myers Squibb, Roche, Janssen, Bayer, AbbVie; Grant/Research Support: 

Boehringer-Ingelheim, AbbVie 

The following authors have nothing to disclose: Adélia Simão, Miriam Santos, 

Raquel S. Silva, Pedro Correia, Lurdes Correia, Pedro Henriques Abreu, Alberto 

Cardoso, José N. Costa 

474 

Long-term Survival Analysis of Re-resection Versus 

Re-radiofrequency Ablation for Intrahepatic Recurrence 

After Initial Hepatectomy or Radiofrequency Ablation 

for Hepatocellular Carcinoma: A 10-year Observational 

Study 

Minjong Lee 1 , Sohee Oh 2 , YOUNG CHANG 1 , Joon Yeul Nam 1 , 

Young Youn Cho 1 , Jeong-Ju Yoo 1 , Yuri Cho 1 , Donghyeon Lee 1 , 

Eun Ju Cho 1 , Jeong-Hoon Lee 1 , Su Jong Yu 1 , Nam-Joon Yi 3 , Jeong 

Min Lee 4 , Kwang-Woong Lee 3 , Jung-Hwan Yoon 1 , Kyung-Suk 

Suh 3 , Yoon Jun Kim 1 ; 1 Department of Internal Medicine and Liver 

Research Institute, Seoul National University College of Medicine, 

Seoul, Korea (the Republic of); 2 Department of Biostatistics, Seoul 

Metropolitan Government Seoul National University Boramae 

Medical Center, Seoul, Korea (the Republic of); 3 Department of 

Surgery, Seoul National University College of Medicine, Seoul, 

Korea (the Republic of); 4 Department of Radiology, Seoul National 

University College of Medicine, Seoul, Korea (the Republic of) 

Background/Aim: Survival outcomes of resection (OP) and 

radiofrequency ablation (RFA) were not significantly different 

in HCC patients with the Barcelona Clinic Liver Cancer (BCLC) 

stage 0/A. However, in patients with recurrent HCC, survival 

rates of patients treated with re-OP have not been compared 

with those with re-RFA. We aimed to compare survival of patient 

treated with re-OP to those with re-RFA in patients with BCLC 

0/A. Methods: This retrospective study included 225 patients 

with recurred BCLC 0/A HCC in Child-Pugh class A/B. According 

to initial treatment strategies, patients were divided into two 

groups; 148 patients treated with RFA; 77 patients with OP. 

For treating recurrent HCC, patients were also divided into two 

treatments: 181 patients treated with RFA; 44 patients with OP. 

Survival rates comparing each group according to treatment 

strategies were analyzed after matching of propensity scores 

after matching of propensity scores according to established 

liver cirrhosis or HCC risk factors. Results: The median tumor 

size 2.4/1.5 cm; single HCC 80.9/84.9% in patients with 

initial/recurrent HCC, respectively. The 5-year overall survival 

rates were 85.3%, 89.5%, 93.0%, and 95.0% in patients 

treated with RFA-RFA, OP-RFA, RFA-OP, and OP-OP for initial 

and recurrent HCC, respectively. Patients treated with re-OP 

for initial and recurrent HCCs showed significantly lower mortality 

than those with re-RFA; hazard ratio (HR)=0.143 (95% 

confidence interval[CI], 0.021–0.991, P=0.04). Patients who 

received resection at least for initial or recurrent HCC showed 

lower mortality than those who only treated with re-RFA; 

HR=0.414 (95% CI, 0.187–0.913, P=0.029). Conclusions: In 

patients with initial and recurrent BCLC stage 0/A, and early 

cirrhosis, who recurred HCC with BCLC stage 0/A, re-hepatectomy 

improves the patient survival compared to those treated 

with re-RFA treatment. Treatments including resection for initial 

or recurrent HCC also improves the survival when compared 

to RFA only treatment. Therefore, for selected patients, re-hepatectomy 

should be considered as the first treatment option for 

initial and recurrent HCC. 

Disclosures: 

Jeong Min Lee - Advisory Committees or Review Panels: Bayer healthcare 

Kwang-Woong Lee - Grant/Research Support: ChongGeunDang, Astellas, 

GreenCross 






The following authors have nothing to disclose: Minjong Lee, Sohee Oh, YOUNG 

CHANG, Joon Yeul Nam, Young Youn Cho, Jeong-Ju Yoo, Yuri Cho, Donghyeon 

Lee, Eun Ju Cho, Jeong-Hoon Lee, Su Jong Yu, Nam-Joon Yi, Jung-Hwan Yoon, 

Kyung-Suk Suh 

475 

Simple serological tests and Transient Elastography as 

screening tools for clinically significant portal hypertension 

in resectable hepatocellular carcinoma 

Simona Bota, Florian Hucke, Mattias Mandorfer, Remy Schwarzer, 

Philipp Schwabl, Thomas Reiberger, Arnulf Ferlitsch, Michael 

Trauner, Wolfgang Sieghart, Markus Peck-Radosavljevic; Gastroenterology 

and Hepatology, Medical University of Vienna, Vienna, 

Austria 

Background:Hepatocellular carcinoma (HCC) resection is recommended 

by the EASL Guideline as first-line treatment option 

for patients with solitary HCC and very well-preserved liver 

function, defined as normal bilirubin with either hepatic venous 

pressure gradient (HVPG)≤10 mmHg.However HVPG measurements 

are available only in highly specialized Hepatology centers. 

Aim:to assess the diagnostic value of serological scores 

(initially developed for fibrosis evaluation) and transient elastrography 

(TE) as screening tool for prediction of HVPG>10 

mmHg in patients with potentially resectable HCC. Methods: 

Our study included 46 consecutive patients with chronic liver 

disease and potentially resectable (Child-Pugh A,normal bilirubin, 

platelet count≥100000cells/mm 3 and no esophageal 

varices) evaluated in our Hemodynamic Labor.We performed 

HVPG measurements, serological tests and TE in a subgroup 

of patients.Simple serological scores were calculated:Fibrosis 

Index (FI),Forns score and Lok score. According to the newly 

proposed TE quality criteria, ten valid measurements with a 

median liver stiffness value ≥7.1kPa and IQR/median>30% 

were considered poorly reliable and not included for analysis. 

TE cases without 10 valid measurements were also excluded. 

Results:The etiology of liver disease was : alcholic -41.3%, 

viral -39.1%, others etiologies – 19.6%. The presence of 

HVPG>10mmHg was diagnosed in 10/46 patients (21.7%). 

Liver stiffness measurements by TE were available in 67.3%of 

patients. Reliable liver stiffness measurements were obtained 

in 87% of patients. Transient Elastography had a very good 

performace (93.7%) to exclude the presence of CSPH, while 

from the serological tests Fibrosis Index (FI) had the best performance 

(NPV 84.7%)(Table) Conclusions:TE is a very good 

tool to exclude the presence of HVPG>10mmHg in patients 

with potentially resectable HCC.Simple serological tests were 

less accurate and could correctly exclude CSPH in 80-85% of 

cases, but were feasible in all patients.


Disclosures: 


Squibb 

















The following authors have nothing to disclose: Florian Hucke, Remy Schwarzer, 

Philipp Schwabl, Arnulf Ferlitsch 

476 

Pro-angiogenic Tie2-expressing monocytes/TEMs as a 

biomarker of the response to sorafenib in patients with 

advanced hepatocellular carcinoma 

Hirotaka Shoji 1,2 , Sachiyo Yoshio 1 , Yohei Mano 1 , Masaya Sugiyama 

1 , Yoshihiko Aoki 1 , Norio Itokawa 3 , Masanori Atsukawa 3 , 

Yosuke Osawa 4 , Kiminori Kimura 4 , Akinobu Taketomi 2 , Masashi 

Mizokami 1 , Tatsuya Kanto 1 ; 1 The Research Center for Hepatitis & 

Immunology, National Center for Global Health and Medicine, 

Ichikawa, Japan; 2 Department of Gastroenterological Surgery 

1, Hokkaido University Graduate School of Medicine, Sapporo, 

Japan; 3 Division of Gastroenterology, Department of Internal Medicine, 

Nippon Medical School Chiba Hokusoh Hospital, Inzai, 

Japan; 4 Division of Hepatology, Tokyo Metropolitan Cancer and 

Infectious Diseases Center Komagome Hospital, Tokyo, Japan 

Background Sorafenib is a multi-kinase inhibitor that has been 

used for the patients with advanced stages of HCC. The survival 

benefit of sorafenib has been clinically proven, however, 

majority of patients are forced to discontinue the therapy due to 

severe adverse effects. It is anticipated to discover the biomarkers 

for the prediction of sorafenib response, in order to establish 

the tailored therapy by selecting patients promising for the 

outcome. We reported that Tie2-expressing monocytes (TEMs) 

were increased in patients with HCC, the frequency of which 

was correlated with the degree of angiogenesis in the liver. 

We sought to examine the possibility of TEMs as the biomarker 

for the drug response in patients with HCC. Furthermore, in 

order to discover more feasible markers reflecting the response 

to sorafenib, we explore the factors involving in the development 

of Tie2 expression on myeloid lineage cells. Method We 

enrolled 25 patients with advanced HCC treated with sorafenib 

in this study (13 Child-Pugh A and 12 CP-B). The anti-tumor 

effect of sorafenib was evaluated after a month and thereafter 

by dynamic CT and MRI with the criteria of modified RECIST. 

We analyzed the frequency of TEMs (CD14+CD16+TIE2+ 

cells) in the periphery of the patients and its correlation with 

clinical and radiological response to sorafenib. We examined 

41 serum factors by multiplexed analysis in 71 HCC patients 

and healthy donors. Finally, we examined the impact of such 

factors on the differentiation of TEMs from monocytes in vitro. 

Results Among the subjects, 9 patients tolerated the continuation 

of sorafenib therapy. They were categorized into the 

responder group (one partial response and 4 stable disease) 

and non-responder group (4 progressive disease) judged by 

the modified RECIST. In the responders, the pre-treatment frequency 

of TEMs tended to be higher than those in the non-responders 

(6.1±2.2% vs. 3.5±0.9%, P=0.06). Furthermore, the 

frequency of TEMs in the responders was dropped from 6.1% 

to 3.5% after one month. In contrast, TEMs in the non-responders 

did not change after the therapy. In patients with high TEMs 

frequency (>2.75% cut-off), the levels of M-CSF, HGF, Osteopontin, 

and Follistatin were significantly higher than those in 

patients with lesser TEMs (p


m 2 for the men and 77.9 ± 44.3 cm 2 /m 2 for the women. The 

median survival times for the men and women were 12.4 and 

20.1 months, respectively. In the multivariate Cox regression 

analysis, only L3-SMI < 43 cm 2 /m 2 (hazard ratio [HR], 5.17; 

95% confidence interval [CI], 1.81–14.79; p = 0.002) and 

Child-Pugh score (HR, 1.95; 95% CI, 1.19–3.21; p = 0.008) 

were independently associated with mortality in the men only. 

Conclusions: Skeletal muscle depletion was a poor prognostic 

factor in the treatment with sorafenib for advanced HCC. Prevention 

of skeletal muscle depletion may be important for the 

improvement of the prognosis in advanced HCC. 

Disclosures: 

The following authors have nothing to disclose: Takashi Hoshino, Atsushi 

Naganuma, Yuhei Suzuki, Daisuke Uehara, Takahiro Miyoshi, Ken Sato, Satoru 

Kakizaki, Masanobu Yamada, Hitoshi Takagi 

478 

The role of cytokines and adhesion molecules in hepatocellular 

carcinoma to predict survival 

Filiz Akyuz 1 , Raim Iliaz 1 , Umit Akyuz 2 , Derya Duranyildiz 1 , Murat 

Serilmez 1 , Cetin Karaca 1 , Kadir Demir 1 , Fatih Besisik 1 , Sabahattin 

Kaymakoglu 1 ; 1 Istanbul University, Istanbul, Turkey; 2 Fatih Sultan 

Mehmet Educational and research center, Istanbul, Turkey 

Introduction: Hepatocellular carcinoma (HCC) is one of the 

most common cancers worldwide. The prognosis is related to 

stage of tumor and predictors of prognosis may determine the 

management of HCC. Some cytokines and growth factors are 

increased in HCC and these may be related with prognosis of 

the cancer. We aimed to investigate the effects of interleukin 

(IL)-32, IL-1 beta, IL- 18, vascular cellular adhesion molecule-1 

(VCAM-1) and epithelial cell adhesion molecule (EpCAM) on 

the prognosis of patients with HCC. Material and Methods: Fifty 

HCC patients and 19 healthy volunteers as a control group 

were enrolled in this prospective study. All demographic and 

clinical data were analyzed. Serum samples were obtained 

on first admition before any adjuvant and metastatic treatment 

was given. Serum IL-32, IL-1 beta, IL-18, VCAM-1 and EpCAM 

were determined by using ELISA kits. Results: All patients had 

cirrhosis and Child-Pugh stage were as follows: %70 Child 

A, 28% Child B and 2% Child C (78% HBV, 2% HCV, others 

cryptogenic). Fifty-eight percent of HCC patients were died 

in median 7.3 months. The mean age of the patients was 

60.2±9.4 years and 53.4±8.2 years for the controls. Mean 

serum level of IL-32 was higher in patients with HCC than controls 

(65.1 vs 13.7 pg/mL) (p0.05). Conclusion: 

IL-32, IL-18, VCAM-1 and EpCAM were significantly higher in 

HCC patients. Although, cytokines can be a diagnostic marker 

for HCC, they had no prognostic value in HCC patients. On 

the other hand EpCAM can be use to the determine prognosis 

of HCC and the management of treatment. 

Disclosures: 

The following authors have nothing to disclose: Filiz Akyuz, Raim Iliaz, Umit 

Akyuz, Derya Duranyildiz, Murat Serilmez, Cetin Karaca, Kadir Demir, Fatih 

Besisik, Sabahattin Kaymakoglu 

479 

Reappraisal of portal vein tumor thrombosis as a prognostic 

factor for patients with hepatocellular carcinoma: 

tumor factor matters 

YOUNG CHANG 1 , Minjong Lee 1 , Joon Yeul Nam 1 , Hongkeun 

Ahn 1 , Hyeki Cho 1 , Yuri Cho 1 , Jeong-Ju Yoo 1 , Donghyeon Lee 1 , 

Young Youn Cho 1 , Eun Ju Cho 1 , Jeong-Hoon Lee 1 , Yoon Jun Kim 1 , 

June Sung Lee 2 , Jung-Hwan Yoon 1 , Su Jong Yu 1 ; 1 Department 

of Internal Medicine and Liver Research Institute, Seoul National 


2 Department of Internal Medicine, Ilsan Paik Hospital, Inje University 

College of Medicine, Goyang, Korea (the Republic of) 

Background/Aim: The presence of portal vein tumor thrombosis 

(PVTT) is a poor prognostic factor for patients with hepatocellular 

carcinomas (HCC). HCC patients with PVTT are considered 

advanced stage regardless of the tumor factor and only 

sorafenib is recommended for those patients by current guidelines. 

This study was aimed to assess whether HCC patients 

with PVTT could have better prognosis with proper treatment if 

the tumor factors were favorable. Methods: This single-center 

retrospective study involved 1,184 patients diagnosed HCC 

from January 2005 to December 2006, before sorafenib 

era. Overall survival (OS) was estimated by the Kaplan-Meier 

method and Cox proportional hazard model. To control the 

different distributions of covariates of the two groups with or 

without PVTT, matched patients were selected by propensity 

score analysis. Results: Among the patients with PVTT, the proportion 

of diffuse infiltrative type was significantly higher than 

nodular type (P


480 

Value of PIVKA II and AFP serum level kinetics for treatment 

monitoring in patients with hepatocellular carcinoma 

(HCC): a French pilot study 

Audrey Payance 1 , Mohamed Bouattour 1 , Mohamed Achahboun 2 , 

Miguel Albuquerque 2 , Laurent Castera 1 , Pierre Bedossa 2 , Olivier 

Soubrane 3 , Francois Durand 1 , Valerie Paradis 2 ; 1 HEPATOLOGY, 

BEAUJON HOSPITAL, Clichy, France; 2 Pathology, BEAUJON 

HOSPITAL, Clichy, France; 3 HPB Surgery and Liver Transplan, 

BEAUJON HOSPITAL, Clichy, France 

Background and Aims: In Asia, prothrombin induced by vitamin 

K absence-II (PIVKA-II) is largely used as a diagnostic and 

prognostic biomarker for hepatocellular carcinoma (HCC). 

We already confirmed the diagnostic value of PIVKA-II for 

European patients with early HCC. Herein, our aim was to 

study the dynamics of PIVKA-II serum level in the follow-up of 

European patients with HCC, according to treatment monitoring. 

Material and Methods: Patients with histologically proven 

HCC diagnosed in our center from January 2013 to August 

2014 were included. For each patient, serum PIVKA-II and 

afetoprotein (AFP) levels were assessed with an enzyme-linked 

immunosorbent assay at two different times. We defined two 

groups of patients: 1) “treated patients” group with first serum 

sample obtained within two months before treatment (D0) and 

the second within 2-3 months after treatment initiation (M2-3), 

2) “non-treated patients” as a control group, with two serum 

samples, at diagnosis (T1) and 2-3 months subsequently (T2). 

Baseline serum levels of AFP and PIVKA-II and dynamics during 

follow-up were compared between the two groups. Results are 

presented as median (interquartile range). Results: A total of 

32 patients with HCC developed on chronic liver disease were 

included (men, 85 %; median age, 59 years; BCLC stage: A 

47% - B 28%- C 19% - D 6%). Aetiologies of underlying liver 

disease were alcohol consumption, viral hepatitis B, viral hepatitis 

C, non alcoholic steato-hepatitis and genetic hemochromatosis 

in 14 (44%) ; 8 (25%), 13 (41%), 4 (12%), 2 (6%) of 

patients respectively. In 23 patients, HCC treatment was performed 

including chemoembolization (n=14), radiofrequency 

ablation (n=3), surgical resection (n=1), sorafenib (n=4), and 

radioembolisation (n=1). For all patients, median baseline AFP 

and PIVKA II serum levels were 11.6 ng/ml (4.8 - 139.8 ng/ 

ml) and 416 mAU/ml (100.5 - 5894.25 mAU/ml), respectively. 

In the “treated patients” group, PIVKA II serum levels 

decreased statistically significantly from D0 to M2-3 (D0: 811 

mAU/ml (100.5-11559.75) vs M2-3: 292 mAU/ml (46.25 

- 7391); p = 0.031)). However, no significant decrease was 

observed regarding AFP serum levels between D0 and M2-3 

(D0: 18.9 ng/ml (4.8-392) vs M2-3: 16.3 ng/ml (4.5-103); 

p=0.831). Finally, no changes of AFP and PIVKA II serum levels 

from baseline T1 to T2 were observed in the control group. 

Conclusions: Significant decrease of PIVKA II serum levels following 

treatment of HCC was observed. These results suggest 

that this biomarker, in contrast to AFP, could be useful for monitoring 

treatment in patients with HCC. Further investigations are 

needed to validate these results. 

Disclosures: 

Mohamed Bouattour - Advisory Committees or Review Panels: Bayer Schering 

Pharma; Speaking and Teaching: Bayer Schering Pharma 

Laurent Castera - Speaking and Teaching: Gilead, BMS, Janssen, Echosens, 

Abbvie, Biopredictive 



The following authors have nothing to disclose: Audrey Payance, Mohamed 

Achahboun, Miguel Albuquerque, Pierre Bedossa, Olivier Soubrane, Valerie 

Paradis 

481 

Liver-directed (LD) Therapy for Intermediate Stage 

Hepatocellular Carcinoma: Survival Across Treatment 

Strategies 

Alvaro E. Castillo 1 , Willscott E. Naugler 2 , Kenneth J. Kolbeck 3 , 

Khashayar Farsad 3 , Susan L. Orloff 1 , Kevin Billingsley 4 , C. Kristian 

Enestvedt 1 ; 1 Surgery, Oregon Health and Science University, 

Portland, OR; 2 Internal Medicine, Oregon Health and Science 

University, Portland, OR; 3 Dotter Interventional Institute, Oregon 

Health and Science University, Portland, OR; 4 Division of Surgical 

Oncology, Oregon Health and Science University, Portland, OR 

Treatment strategies favor LD options for BCLC intermediate 

stage HCC. Evaluate the outcomes of different multidisciplinary 

treatment strategies in BCLC B patients. A retrospective cohort 

analysis of HCC patients referred to our institutional Multidisciplinary 

Liver Tumor Board. All with Barcelona Clinic Liver 

Cancer (BCLC) stage B who underwent either trans-arterial 

chemoembolization (TACE), radiation therapy (XRT)/TACE, 

Yttrium-90 (Y-90) alone and Y-90/TACE were selected. Resection, 

ablation or transplantation was excluded. Univariate analysis 

was used to compare demographics, tumor characteristics, 

and treatment modalities. Multivariate analysis of clinical factors 

associated with survival was performed using Cox proportional 

hazards modeling. Survival was compared using the 

Kaplan Meier log-rank method. Eighty eight BCLC B patients 

were included. Comparison by treatment groups showed 

no difference in severity of disease or demographics except 

for bilirubin which was higher for the TACE and TACE/XRT 

groups when compared to Y-90 and Y-90/TACE (p=0.024). 

Those who received Y-90 had larger tumors (median 6.5 cm, 

p=


The following authors have nothing to disclose: Alvaro E. Castillo, Willscott 

E. Naugler, Kenneth J. Kolbeck, Susan L. Orloff, Kevin Billingsley, C. Kristian 

Enestvedt 

482 

Analyses of progressive disease patients with advanced 

hepatocellular carcinoma treated with sorafenib: prognosis, 

treatment, and progression pattern 

Keiko Yamane, Takahide Nakazawa, Wasaburo Koizumi; Gastroenterology, 

Kitasato University School of Medicine, Sagamihara, 

Japan 

Purpose: Progressive disease (PD) is highly observed in patients 

with advanced hepatocellular carcinoma (HCC) treated with 

sorafenib. Here, we examined PD patients about progression 

pattern, treatment for PD, and the prognosis. Methods: PD 

patients radiologically assessed using RECIST 1.1. were retrospectively 

examined. Overall survival (OS), Time to progression 

(TTP), post-progression survival (PPS), progression pattern, 

and prognostic factors were analyzed. Progression pattern was 

classified as follows (i-v): (i) intrahepatic increase in tumor size 

(ii) extrahepatic increase in tumor size (iii) new intrahepatic 

lesion (iv) new extrahepatic lesion (v) new intra/extrahepatic 

lesion. Result: One-hundred and eight patients were treated 

with sorafenib, and 102 were radiologically assessed. PD was 

observed in 56 patients (median age 69 year old, hepatitis 

C virus 50%, Child-Pugh [C-P] A 98%, and BCLC-C 61%). 

Twenty-four patients of PD (43%) had invasion to intrahepatic 

major vessels. The median OS, TTP, and PPS were 7, 1.9, 

and 5 months, respectively. Progression pattern was (i) in 12 

patients, (ii) in 1, (iii) in 30, (iv) in 6, (v) in 7. The median OS 

by progression pattern was 7.5 months in (i), 15 in (ii), 7.1 in 

(iii), 6.9 in (iv), and 2.6 in (v). The pattern of new intra/extrahepatic 

lesion showed significantly poor prognosis. C-P score 

of 5 point (P = 0.004, OR 0.402, 95% CI 0.214-0.753) and 

invasion to intrahepatic major vessels (P = 0.017 OR 2.119, 

95% CI: 1.142-3.932), at the beginning of sorafenib treatment 

were independent prognositic factors of OS by a Cox proportion 

hazard model. While, C-P A (P = 0.036, OR 0.450, 95% 

CI 0.213-0.947), invasion to vessels (P = 0.008 OR 2.605, 

95% CI: 1.278-5.306) and new intra/extrahepatic lesion (P = 

0.004, OR 4.517, 95% CI: 1.618-12.608) were independent 

factors of PPS by a Cox model. Second line treatment for PD 

determined the good prognosis. Median PPS was 6.2 months 

in PD patients with 2nd line treatment, while median PPS of 2.5 

months without any treatment. C-P A of hepatic function was 

statistically observed in PD patients with 2nd line treatment by 

Logistic-regression analysis (P = 0.046, OR 4.317, 95% CI 

0.215-18.183). Conclusion: Hepatic function and progression 

pattern were important factors to determine the prognosis in PD 

patients. Second line treatment based on these findings should 

be considered. 

Disclosures: 

The following authors have nothing to disclose: Keiko Yamane, Takahide Nakazawa, 

Wasaburo Koizumi 

483 

Impact of Viral Hepatitis Etiology on Survival Outcomes 

in Patients with Hepatocellular Carcinoma: A Large Multicentre 

Study 

Sara Mgaieth 1 , William W. Kemp 1 , Paul Gow 2 , Michael A. Fink 3 , 

Virginia H. Knight 4 , Anouk T. Dev 4 , Amanda J. Nicoll 5 , John 

Lubel 5 , Sally Bell 6 , Thai Hong 6 , Marno C. Ryan 6 , Siddharth Sood 7 , 

Stuart K. Roberts 1 ; 1 Gastroenterology, The Alfred Hospital, Melbourne, 

VIC, Australia; 2 Liver Transplant Unit, The Austin Hospital, 

Melbourne, VIC, Australia; 3 Department of Surgery, The Austin 

Hospital, Melbourne, VIC, Australia; 4 Gastroenterology, Monash 

Medical Center, Melbourne, VIC, Australia; 5 Gastroenterology, 

Eastern Health, Melbourne, VIC, Australia; 6 Gastroenterology, St 

Vincent’s Hospital, Melbourne, VIC, Australia; 7 Gastroenterology, 

The Royal Melbourne Hospital, Melbourne, VIC, Australia 

BACKGROUND: Hepatocellular carcinoma is the fifth most common 

cancer worldwide in men, and the second leading cause of 

death. The incidence of HCC is increasing, with HBV and HCV 

infections accounting for more than 50% of cases. Although 

HBV and HCV are known risk factors for HCC it is unclear if 

there is an overall survival difference between patients with 

viral and non-viral hepatitis related HCC. AIM: The aim of the 

study is to compare the overall survival and recurrence free survival 

of patients with viral and non-viral hepatitis related HCC 

and between those with viral hepatitis B and C related HCC. 

METHODS: A multi-centre study was conducted on HCC cases 

from 6 large tertiary hospitals from 1994-2014. Patient demographics, 

severity of liver disease, tumour characteristics, and 

patient outcomes were retrieved from the HCC database and 

computer records. Survival outcomes were compared between 

the three cohorts of HBV, HCV and non-viral related HCC and 

predictors of survival were determined using Cox proportional 

hazards regression. RESULTS: There were a total of 1551 cases 

of HCC between 1994-2014. 17% (265) had HBV, 35% 

(541) had HCV, 30 (3%) co-infected. There was a total of 50% 

(776) with viral hepatitis related HCC, and 43% (661) with 

non-viral hepatitis related HCC, and 7%(114) unknown. On 

univariate analysis, patients with viral hepatitis related HCC 

had better overall survival compared to those with non-viral 

hepatitis related HCC (P


The following authors have nothing to disclose: Sara Mgaieth, William W. 

Kemp, Paul Gow, Michael A. Fink, Virginia H. Knight, Anouk T. Dev, John Lubel, 

Thai Hong, Marno C. Ryan 

484 

Stereotactic body radiation therapy for recurrent hepatocellular 

carcinoma as a local salvage treatment 

Baek Gyu Jun 1 , Sae Hwan Lee 1 , Hong Soo Kim 1 , Sang Gyune 

Kim 3 , Young Seok Kim 3 , Boo Sung Kim 3 , Soung Won Jeong 4 , 

Jae Young Jang 4 , Young Don Kim 2 , Gab Jin Cheon 2 ; 1 Department 

of Internal Medicine, Soonchunhyang University College of 

Medicine Cheonan Hospital, Cheonan-si, Korea (the Republic of); 

2 Internal Medicine, University of Ulsan College of Medicine, Gangneung, 

Korea (the Republic of); 3 Department of Internal Medicine, 

Soonchunhyang University College of Medicine Bucheon Hospital, 

Bucheon, Korea (the Republic of); 4 Department of Internal Medicine, 

Soonchunhyang University College of Medicine Seoul Hospita, 


BACKGROUND: The aim of this study was to evaluate the 

efficacy and toxicity of stereotactic body radiation therapy 

(SBRT) as a local salvage treatment for small, recurrent hepatocellular 

carcinoma(HCC). METHODS: Between March 2011 

and February 2014, 32 patients with recurrent HCC(diameter 


486 

Clinical Features and Treatment Modalities of Long-Term 

Survivors with Advanced Hepatocellular Carcinoma: A 

Single Institution Cohort Study 

Kyung Hee Kim 1 , Joong-Won Park 1,2 , Bo Hyun Kim 1 , Chang-Min 

Kim 1 , Byung Ho Nam 2 ; 1 Center for Liver Cancer, National cancer 

center, Go yang si, Korea (the Democratic People’s Republic of); 

2 Department of Cancer Control and Policy, Graduate School of 

Cancer Science and Policy, National Cancer Center, Go yang si, 

Korea (the Democratic People’s Republic of) 

Background and Aim: The prognosis of patients with advanced 

hepatocellular carcinoma (HCC) is poor despite systemic or 

local treatments. We analyzed the characteristics of long-term 

survivors (LTS) with advanced HCC to determine the prognostic 

and treatment factors responsible for long-term survival. Methods: 

Between January 2001 and December 2010, 543 patients 

were first diagnosed as having modified International Union 

Against Cancer (mUICC) stage IV HCC, with Child-Pugh (CP) 

class A liver function at the National Cancer Center, Korea. 

The authors retrospectively reviewed all medical records and 

radiographs. Among the patients, we compared clinical features 

and treatment modalities of the LTS group (survival > 24 

months; n = 66) with those of the control (C) group (survival < 6 

months; n = 333). Results: For 543 patients, the median overall 

survival (OS) was 4.6 months (95% CI: 4.2-5.0): 37.7 months 

(95% CI: 34.4-41.0) in the LTS group and 3.1 months (95% 

CI: 2.9-3.3) in the C group. Compared to the C group, LTS 

group patients were significantly more likely to be older and 

to have non-hepatitis B virus etiology, lower tumor numbers, 

smaller tumor size, nodular-type tumors, absence of portal vein 

invasion, and better liver function. Ill-defined tumor type, portal 

vein invasion and CP A6 proved to be independent prognostic 

variables (p


Patients characteristics 

fatty liver disease. Overall, the percentage of non-cirrhotic, 

non-HBV HCC was 2.93%, non-cirrhotic, non-HBV HCC with 

hepatitis C was 1.66%, and non-cirrhotic, non-HBV HCC with 

fatty liver disease was 6.6%. Conclusions Non-HBV HCC can 

arise in the setting of chronic hepatitis C and fatty liver disease 

without advanced fibrosis or cirrhosis. Our prevalence rates 

may underestimate the true prevalence owing to the exclusion 

of patients without adequate histology available. 

Disclosures: 

Steven-Huy B. Han - Grant/Research Support: Bristol Myers Squibb, Gilead 

The following authors have nothing to disclose: Vivian Ng, Jennifer Phan, Alan 

J. Sheinbaum 

Disclosures: 

Rene Adam - Grant/Research Support: Merck Serono, Roche, Sanofi aventis, 

astellas, novartis, Merck Serono, Roche, Sanofi aventis, astellas, novartis, Merck 

Serono, Roche, Sanofi aventis, astellas, novartis, Merck Serono, Roche, Sanofi 

aventis, astellas, novartis 



The following authors have nothing to disclose: Eleonora De Martin, Michel 

Rayar, Damien Bergeat, Luis Filipe Abreu de Carvalho, Maximiliano Gelli, Denis 

X. Castaing, Daniel Cherqui, Antonio Sa-Cunha, Emmanuel Boleslawski, Karim 

Boudjema, Eric Vibert 

488 

Non-Cirrhotic Hepatocellular Carcinoma in the Absence 

of Hepatitis B 

Vivian Ng 1 , Jennifer Phan 1 , Alan J. Sheinbaum 2 , Steven-Huy B. 

Han 1 ; 1 Pfleger Liver Institute, University of California, Los Angeles, 

Los Angeles, CA; 2 Gastroenterology, VA West Los Angeles Healthcare 

Center, Los Angeles, CA 

Purpose Hepatocellular carcinoma (HCC) accounts for approximately 

90% of primary hepatic malignancies worldwide. 

In hepatitis B virus (HBV) infection, HCC can develop in the 

absence of cirrhosis. However, it is thought that in chronic 

hepatitis C virus infection and fatty liver disease, development 

of cirrhosis is necessary prior to formation of HCC. To date, 

there have only been a small number of case reports showing 

that HCC can develop in patients with chronic hepatitis C and 

in fatty liver disease without evidence of underlying cirrhosis. 

We sought to evaluate whether or not non-cirrhotic HCC exists 

and assess underlying risk factors contributing to its development. 

Methods We performed a single-center, retrospective 

chart review study at a large academic tertiary care hospital. 

All patients seen in both the liver cancer clinic and orthotopic 

liver transplantation (OLT) clinic for liver cancer in the last 10 

years were evaluated. We determined how many patients 

had non-cirrhotic HCC based on available pathology reports, 

such as explanted liver pathology, liver resection pathology, 

and liver biopsy pathology. Patients with hepatitis B, non- 

HCC tumors, and patients without both tumor and non-tumor 

liver histology available were excluded. Demographic information, 

pertinent lab values, and co-morbid conditions were 

recorded. Results 1925 unique patients were identified. 1382 

patients were excluded from the study, including 303 patients 

with hepatitis B, 510 patients with non-HCC tumors, and 569 

patients without available tumor and non-tumor histology. Of 

the remaining patients, 491 patients underwent (OLT), 70 

underwent local ablative therapy, 42 underwent resection of 

the HCC, and 17 underwent systemic therapy. All 491 OLT 

patients had underlying cirrhosis. Of the 134 patients who 

did not undergo (OLT), 35 patients (26.1%) were identified as 

having non-cirrhotic HCC. 13 patients (37.1%) had an underlying 

diagnosis of hepatitis C, and 6 patients (17.1%) had 

489 

Albi score predicts survival in patients with BCLC 0/A 

stage Hepatocellular Carcinoma (HCC) independently of 

Child Pugh (CP) score and treatment allocation 

Simona Onali 1 , Aileen Marshall 1 , Dinesh Sharma 1 , Pam O’Donoghue 

1 , Emily Dannhorn 1 , Phillip Johnson 2 , James O’Beirne 1 ; 

1 Sheila Sherlock Liver Centre, Royal Free Hospital, London, United 

Kingdom; 2 Department of Molecular and Clinical Cancer Medicine, 

University of Liverpool, Liverpool, United Kingdom 

Introduction: BCLC 0/A stage HCC patients are considered 

candidates for curative treatments. Within the BCLC classification, 

liver function is assessed by CP score despite this score 

not being derived from patients with HCC. ALBI is a novel 

evidence based score derived from a large cohort of HCC 

patients and has been shown to define different prognostic 

groups even within CP A. 1 We studied the performance of ALBI 

score in a well characterised cohort of patients with BCLC 0/A 

stage HCC to determine if ALBI score impacted on prognosis. 

Methods: Consecutive patients with BCLC 0/A HCC, evaluated 

between 2010-2014 at the Royal Free Hospital were included. 

All patients had a performance score of 0 and were undergoing 

evaluation for potential resection. Patients were considered 

resectable if they had HVPG≤10mmHg or HVPG≥10mmHg 

with good liver function as assessed by ICG clearance. ALBI 

score was calculated at the time of HVPG and ICG clearance 

measurement and divided into 3 grades as per published cutoffs. 

Kaplan Meier and Cox regression analysis were used to 

assess the impact of ALBI on survival. Results: 67 patients were 

included: 64 (95.5%) were CP A, 3 (4.5%) were CP B. 60 

(89%) had uninodular tumour and 7 (10.4%) multifocal (≤3 

tumours ≤30mm). 46 (69%) underwent resection, 4 (6%) RFA, 

9 (14%) TAE, 3 (5%) OLT, 3 (5%) either no treatment or undergoing 

evaluation. Median follow up was 9 months (13-54). 

10 (15%) patients died during follow up. Cause of death was 

HCC-related n= 4, liver failure n=4, non-liver related n=2. In 

Kaplan Meier analysis CP score was not associated with difference 

in survival. Cox regression analysis revealed ALBI score as 

the only significant factor related to survival and was independent 

of CP score and treatment allocation (p=0.002, OR=15, 

95%CI=3-80). Of patients with ALBI grade 1, 44 (83%) were 

considered suitable and underwent resection vs only 2 (14%) 

with ALBI 2. One-year survival was 88% in patients with ALBI 

grade 1 and 43% in patients with ALBI grade 2 (p=0.002). 

Conclusion: In patients with BCLC 0/A stage HCC ALBI score 

clearly defines patients with different prognosis regardless 

of Child Pugh score and treatment allocation. ALBI grade 2 

patients are very rarely suitable candidates for resection. ALBI 

score is more informative than Child Pugh score in patients with 

early stage HCC and may be useful in determining treatment 

allocation References: 1 . Johnson PJ et al. Assessment of Liver 

Function in Patients With Hepatocellular Carcinoma: A New


Evidence-Based Approach-The ALBI Grade. J Clin Oncol. 2015 

Feb 20;33(6):550-8 

Disclosures: 

The following authors have nothing to disclose: Simona Onali, Aileen Marshall, 

Dinesh Sharma, Pam O’Donoghue, Emily Dannhorn, Phillip Johnson, James 

O’Beirne 

490 

Albi score predicts survival independently of hepatic 

venous pressure gradient (HVPG) and indocyanine 

green (ICG) clearance in HCC patients undergoing resection 

Aileen Marshall 1 , Simona Onali 1 , Dinesh Sharma 1 , Pam O’Donoghue 

1 , Emily Dannhorn 1 , Phillip Johnson 2 , James O’Beirne 1 ; 

1 Sheila Sherlock Liver Centre, Royal Free Hospital, London, United 

Kingdom; 2 Department of Molecular and Clinical Cancer Medicine, 

University of Liverpool, Liverpool, United Kingdom 

Introduction: A new evidence based model, the Albumin-Bilirubin 

(ALBI) grade † , has been recently proposed for assessing 

liver function in patients with hepatocellular carcinoma 

(HCC). We aimed to evaluate the impact of ALBI in predicting 

decompensation and survival after liver resection (LR) for 

HCC. Method: Consecutive patients undergoing resection for 

HCC between 2011-2014 at the Royal Free Hospital were 

evaluated. Demographic, clinical data and histopathological 

features of resected tumour were collected. Patients underwent 

HVPG and ICG clearance measurement to evaluate potential 

resectability. ALBI score was calculated pre and post-operatively. 

Patients were divided into 3 categories (ALBI 1,2,3) 

according to published ALBI score cut-offs. Cox regression was 

used to identify predictors of decompensation and survival post 

LR. Results: 48 patients were included with a median post LR 

follow up 17 months (1-54): male 42(87.5%), mean age 63 

years (28-83). Median HVPG 6 mmHg(2-15), median ICG PDR 

18.2 (6.1-29.4), median ICG R15 6.5 (1.2-19.2). All patients 

had a Child Pugh score A5. 46 (96%) had a pre-LR ALBI grade 

1 and 2 (4%) had an ALBI grade 2. Clinically significant portal 

hypertension (HVPG ≥10mmHg) was found in 11 (23%) 

patients. Eight (19%) patients had an ICG PDR 15. Thirty (62,5%) patients underwent 

an anatomical resection, while 18 (37,5%) had a wedge 

resection. Post-LR decompensation was observed in 5(10%) 

patients: ascites n=4, encephalopathy n=1 at a median of 10 

days post-LR. 12(25%) patients had HCC recurrence after a 

mean time of 11 months (5-28). 7 (15%) died after a mean 

follow up of 15 months (1-40), 3 of them due to tumour recurrence. 

Patients with 1 HCC


Inherited severe unconjugated hyperbilirubinemia is known as 

Crigler-Najjar syndrome (CN). CN is characteristically caused 

by deficiency of UDP-glucuronosyl transferase 1A1 (UGT1A1), 

encoded by the UGT1A1 gene. This enzyme is responsible 

for glucuronidation of bilirubin needed to excrete this toxic 

compound via the bile. The severity of inherited unconjugated 

hyperbilirubinemia depends on the residual UGT1A1 activity 

and can be described as a spectrum from complete absence 

(CN type 1) or some activity (CN type 2) to 30% of residual 

activity as seen in the benign Gilbert syndrome. In CN 

patients with residual UGT1A1 activity a reduction of serum 

bilirubin levels can be achieved by phenobarbital administration 

that results in transcriptional induction of UGT1A1 via 

activation of the Constitutive Androstane Receptor (CAR). We 

analyzed the UGT1A1 promoter sequence of a patient with 

severe unconjugated hyperbilirubinemia suspected for CN type 

2, but unresponsive to phenobarbital treatment and without 

clarifying mutation in the UGT1A1 coding region. Besides an 

extra TA repeat in the TATA box, we disclosed a homozygous 

three nucleotide (3nt) insertion on position -85 to -83 of the 

proximal promoter, corresponding with the HNF1α binding 

site. We generated UGT1A1 promoters with all different mutations 

and cloned them upstream of a luciferase reporter gene 

to perform functional studies. The insertion of this 3nt in the 

HNF1α binding site resulted in a >95% reduction of promoter 

activity, independent of the extra TA-repeat in the TATA-box. In 

addition, this insertion rendered the promoter unresponsive to 

the induction by phenobarbital via CAR and rifampicin via the 

Pregnane X Receptor (PXR), thereby explaining the intermediate 

CN phenotype of this patient. In conclusion, disruption of the 

HNF1α binding site in the promoter of a liver specific gene 

such as UGT1A1 is for the first time demonstrated to have a 

major impact on gene expression in man and may result in 

severe liver disorder. 

Disclosures: 







The following authors have nothing to disclose: Remco van Dijk, Isabel 

Mayayo-Peralta, Sem J. Aronson, Anja A. Kattentidt-Mouravieva, Vincent van 

der Mark, Nevin Oruç, Piter J. Bosma 

493 

MicroRNA-122 (miR-122) Regulates Polyploidy in the 

Liver 

Shu-hao Hsu 1 , Evan Delgado 1 , P. A. Otero 1 , Kolin Meehan 1 , Justin 

B. Moroney 1 , Kun-Yu Teng 2 , Kalpana Ghoshal 2 , Andrew W. 

Duncan 1 ; 1 Department of Pathology, McGowan Institute for Regenerative 

Medicine, University of Pittsburgh, Pittsburgh, PA; 2 Department 

of Pathology, Comprehensive Cancer Center, The Ohio State 

University, Columbus, OH 

A defining feature of the mammalian liver is polyploidy, a 

numerical change in the entire complement of chromosomes. 

Hepatocytes are either mononucleate or binucleate, and 

ploidy is determined by the number of nuclei/cell, as well as 

the ploidy of each nucleus. The first step of polyploidization 

involves cell division with failed cytokinesis. Although polyploidy 

is common, affecting ~90% of hepatocytes in mice and 

50% in humans, the specialized role played by polyploid cells 

in liver homeostasis and disease remains poorly understood. 

The goal of this study was to identify novel signals that regulate 

polyploidization, and we focused on microRNAs (miR- 

NAs). First, to test whether miRNAs could regulate hepatic 

polyploidy we examined livers from Dicer1 knockout mice, 

which are devoid of mature miRNAs. Loss of miRNAs resulted 

in a 3-fold reduction in binucleate hepatocytes, indicating that 

miRNAs could indeed regulate polyploidization. Secondly, we 

surveyed age-dependent expression of >500 miRNAs (NanoString) 

in wild type mice and identified a subset of miRNAs, 

including miR-122, differentially expressed at 2-3 weeks, a 

period when extensive polyploidization occurs. Thirdly, we 

examined Mir122 knockout mice and observed profound, lifelong 

depletion of polyploid hepatocytes, proving that miR-122 

is required for complete hepatic polyploidization. Next, we 

identified direct targets of miR-122, Cux1, Iqgap1, Mapre1, 

Nedd4l and Slc25a34, that regulate cytokinesis. Inhibition of 

each target induced cytokinesis failure and promoted hepatic 

binucleation. Finally, to validate the relevance of these targets 

to liver disease, we examined expression in a subset of human 

hepatocellular carcinomas (HCC) with reduced miR-122. Consistent 

with the mouse data, target expression was inversely 

proportional to miR-122. In summary, our data suggest a novel 

regulatory role for miR-122 in liver polyploidization. Moreover, 

differential expression of miR-122 targets in HCC provides new 

insights into miR-122-mediated tumorigenesis. These studies 

will serve as the foundation for future work investigating miR- 

122 and polyploidy in lipid metabolism, viral infection, tumorigenesis 

and regeneration. 

Disclosures: 

The following authors have nothing to disclose: Shu-hao Hsu, Evan Delgado, P. 

A. Otero, Kolin Meehan, Justin B. Moroney, Kun-Yu Teng, Kalpana Ghoshal, 

Andrew W. Duncan 

494 

Overexpression Screening to Discover Gene-Gene Interactions 

that Drive Hepatocellular Carcinoma 

Monica Teta-Bissett 1 , Kirk J. Wangensteen 1,2 , Klaus H. Kaestner 1 ; 

1 Genetics, University of Pennsylvania, Philadelphia, PA; 2 Gastroenterology, 

University of Pennsylvania, Philadelphia, PA 

Hepatocellular carcinoma (HCC) is the third leading cause 

of cancer death worldwide, and one of the top ten in the 

United States according to the Global Cancer Statistics and 

the US CDC, respectively. Sorafenib, a small molecule targeting 

receptor tyrosine kinases, is the only drug approved for 

treatment of advanced HCC in the United States. In order to 

determine the relative contribution of HCC candidate genes in 

HCC formation and growth, we developed an overexpression 

model to screen multiple gene combinations simultaneously 

during liver tumor formation. A pooled plasmid library of 34 

constructs, each designed to express the FAH gene plus a different 

cancer-related gene of interest, was delivered into the 

liver of Fah-null mice by hydrodynamic tail vein injection. The 

constructs allowed for constitutive expression of FAH, rescuing 

the tyrosinemia of Fah-null mice, and creating repopulation 

nodules overexpressing random combinations of the genes 

within the plasmids. The mouse livers were riddled with tumors 

at four and six months following library injection. More than 

20 unique tumors were dissected and the tumor phenotypes 

were characterized using hematoxylin and eosin staining and 

immunohistochemistry for Osteopontin (Opn) on serial sections. 

DNA was extracted from unique tumors and, by high-throughput 

sequencing of unique 5-nucleotide barcodes attached to 

each of the expression plasmids, the profiles of the exogenous 

DNA constructs was compiled. We found that c-myc 

was linked to all of the tumors that were tested, indicating 

that it was a consistent driver of tumor growth in this system. 

TGFα, Foxa3, Akt, Met and Nr1h3 were among other factors 

detected to drive tumor growth. Interestingly, the combination


of four genes, c-myc, AKT, MET and Nr1h3, correlated with a 

subset of Opn-positive HCC tumors. This innovative paradigm 

allows for prospective testing of millions of combinations of 

overexpressed factors to determine the interaction of known 

and suspected liver oncogenes and tumor suppressors in a 

systematic fashion. 

Disclosures: 

The following authors have nothing to disclose: Monica Teta-Bissett, Kirk J. Wangensteen, 

Klaus H. Kaestner 

495 

Peretinoin, an Acyclic Retinoid, Inhibits Hepatocarcinogenesis 

through Suppression of Sphingosine Kinase 1 

Expression In vitro and In vivo 

Masaya Funaki 1 , Tetsuro Shimakami 1 , Tsuguhito Ota 2 , Masao 

Honda 1 , Takayoshi Shirasaki 1 , Shuichi Kaneko 1 ; 1 Disease Control 

and Homeostasis, Kanazawa university, Kanazawa, Japan; 

2 Brain/Liver Interface Medicine Research Center, Kanazawa University, 


[Objective] A sphingolipid, sphingosine-1-phospate (S1P) is 

a potent bioactive lipid which can regulate carcinogenesis 

and cancer progression. Both sphingosine kinase 1 (SPHK1) 

and SPHK2 are the essential kinases that produce S1P. Therefore, 

SPHK can be a therapeutic target by crucially regulating 

sphingolipid metabolism in several kinds of cancer. Peretinoin 

(NIK-333), an acyclic retinoid, was reported to inhibit the post 

therapeutic recurrence of hepatocellular carcinoma (HCC) in 

patients with chronic hepatitis C. However, the mechanism of 

its inhibitory effects against recurrent HCC remains unclear. 

We hypothesized that peretinoin could prevent hepatocarcinogenesis 

by modifying a SPHK-S1P axis. In the present study, 

we assessed the effect of peretinoin on SPHK activation and 

development of liver cancer in vivo and in vitro. [Method] We 

examined the effect of peretinoin on the expression and the 

enzymatic activity of SPHK1 in a human hepatoma cell line, 

Huh-7 cells. Next, using a liver cancer mice model with atherogenic 

high fat (AHF) diet-induced NASH we administrated 

an AHF diet with and without peretinoin (0.03%) for 48 weeks 

and examined the effect of peretinoin on hepatocarcinogenesis 

and expression of SPHK1. Finally, we clarified the effect of 

SPHK1 on hepatocarcinogenesis induced by Diethylnitrosoamine 

(DEN) using SPHK1 knockout mice. [Results] [In vitro] The 

treatment with peretinoin (10 to 40 μM) reduced the mRNA 

and protein expression of SPHK1 in Huh-7 cells in a time- and 

dose- dependent manner. However, peretinoin did not change 

the expression of SPHK2 in Huh-7 cells. Furthermore, peretinoin 

markedly reduced the enzymatic activity of SPHK1 assessed by 

in vitro 32P labeled SPHK activity assay. Next, we performed 

reporter gene assays by using constructs containing the SPHK1 

promoter region. Peretinoin reduced the promoter activity of 

SPHK1, and overexpression of SP1 restored promoter activity. 

In addition, three deletion constructs with and without SP1 

binding sites were created. The deletion constructs without SP1 

binding sites abolished the promoter activity. Interestingly, we 

previously reported that peretinoin suppresses the expression of 

SP1. Collectively, peretinoin reduced the expression of SPHK1 

mRNA via SP1. [In vivo] At week 48 week of treatment, peretinoin 

down-regulated SPHK1 mRNA expression and prevented 

AHF diet-induced hepatocarcinogenesis. The SPHK1 knockout 

mice developed significantly less numbers of HCC induced by 

DEN than did wild-type mice. [Conclusion] Our data indicate 

that peretinoin prevents hepatocarcinogenesis at least partly 

through reduction of expression and activation of SPHK1. 

Disclosures: 






The following authors have nothing to disclose: Masaya Funaki, Tetsuro Shimakami, 

Tsuguhito Ota, Masao Honda, Takayoshi Shirasaki 

496 

Therapeutic overexpression of miR-122 protects mice 

from chronic alcoholic liver injury through regulation of 

hypoxia-inducible factor-1α 

Abhishek Satishchandran 1 , Aditya Ambade 2 , Banishree Saha 2 , 

Benedek Gyongyosi 2 , Patrick Lowe 2 , Nicita Mehta 2 , James V. Zatsiorsky 

2 , Arvin Iracheta-Vellve 2 , Jia Li 3,4 , Donna Catalano 2 , Karen 

Kodys 2 , Li Zhong 3,4 , Jun Xie 3,5 , Shashi Bala 2 , Guangping Gao 3,5 , 

Gyongyi Szabo 2 ; 1 University of Massachusetts Medical School, 

Worcester, MA; 2 Medicine, UMass Medical School, Worcester, 

MA; 3 Gene Therapy Center, UMass Medical School, Worcester, 

MA; 4 Pediatrics, UMass Medical School, Worcester, MA; 5 Microbiology 

and Physiology Systems, UMass Medical School, Worcester, 

MA 

Introduction: miR-122 regulates essential pathways in alcoholic 

liver disease (ALD). We have observed that chronic alcohol 

administration reduces miR-122 levels in murine hepatocytes. 

Given its essential role in hepatic homeostasis, we hypothesized 

that loss of miR-122 contributes to ALD and can be a therapeutic 

target. In this study, our goals were to asses: first, the 

role of miR-122 in the pathogenesis of ALD and the therapeutic 

potential of miR-122 restoration; second, the role of Hypoxia 

Inducible Factor 1-α (HIF-1α), a putative miR-122 target; and 

third, the mechanisms by which alcohol downregulates miR- 

122. Methods: Wild-type 8-week-old, C57Bl/6 or hepatocyte 

specific HIF-1α knockout (hepHIFKO) mice were injected intravenously 

with hepatocyte-tropic AAV (AAV8) containing antimiR-122 

TuD (TuD), or scrambled (Scr) vector. After 14 days, 

mice were started on a Lieber-DeCarli (LDC) alcohol diet (Et) or 

calorie-matched control diet (PF) for 4 weeks. Some WT mice 

were treated with AAV8 pri-miR-122 (OX) on day 7 of the 

LDC diet. Results: Anti-miR-122 TuD treatment alone resulted in 

significant increases in liver injury (ALT), steatosis, inflammation 

(TNFα, IL1-β, MCP-1), and fibrosis (Acta2, pro-col1α) in PF 

mice compared to Scr-treated controls. The co-administration of 

miR-122 TuD and alcohol resulted in a synergistic effect, further 

increasing liver injury, inflammation and fibrosis. Restoration of 

miR-122 in hepatocytes of alcohol-fed mice by treatment with 

scAAV8 OX resulted in a significant improvement in serum ALT, 

inflammation and fibrosis suggesting a protective role for miR- 

122 in ALD. The hepatic expression and DNA-binding activity 

of HIF-1α, a miR-122 target, was increased in TuD+PF mice 

equivalent to that of alcohol feeding alone. HIF1-α activity was 

the highest in TuD+Et mice compared to all other groups. Hep- 

HIF deficiency protected mice from liver damage, inflammation 

and fibrosis induced either by alcohol or miR-122 knockdown. 

We discovered that alcohol specifically inhibits expression 

of the precursor miR-122 transcript (pri-miR-122) at the transcriptional 

level. Furthermore, grainyhead-like 2 (GRHL2), an 

inhibitor of miR-122 transcription, was significantly increased 

in hepatocytes of alcohol fed mice compared to controls. Conclusions: 

Our findings dissect a novel mechanistic regulatory 

axis of miR-122 and demonstrate the therapeutic potential of 

miR-122 restoration in ALD. miR-122 inhibition alone mimics 

and together with alcohol augments the steatosis, inflammation 

and early fibrosis seen in ALD through its downstream target,


HIF-1α. Alcohol inhibits miR-122 via transcriptional repression 

mediated by GRHL2 upregulation. 

Disclosures: 

Guangping Gao - Stock Shareholder: VoyagerTherapeutics 

The following authors have nothing to disclose: Abhishek Satishchandran, Aditya 

Ambade, Banishree Saha, Benedek Gyongyosi, Patrick Lowe, Nicita Mehta, 

James V. Zatsiorsky, Arvin Iracheta-Vellve, Jia Li, Donna Catalano, Karen Kodys, 

Li Zhong, Jun Xie, Shashi Bala, Gyongyi Szabo 

497 

Niclosamide ethanolamine inhibits growth of patient-derived 

hepatocellular carcinoma xenografts 

Bin Chen 1 , Wei Wei 2 , Mei-Sze Chua 2 , Atul Butte 1 , Samuel K. So 2 ; 

1 Institute for Computational Health Sciences, UCSF, San Francisco, 

CA; 2 Asian Liver Center, Stanford University, Stanford, CA 

Background and Aim Hepatocellular carcinoma (HCC) is a fatal 

malignancy with a dismal prognosis. Sorafenib is the only drug 

approved for the treatment of advance HCC patients in the last 

decade; it is associated with only a modest response rate. The 

recent advances in genomics and transcriptomics have led to 

large volumes of molecular data for HCC, providing an unprecedented 

opportunity to translate these data into more effective 

therapeutics. To address the unmet clinical need in HCC, we 

leveraged the public large datasets on disease and drug gene 

expression signatures to identify drug candidates that may 

have yet unidentified anti-tumor efficacy in HCC. Materials and 

Methods We created a HCC gene expression signature by 

analyzing 250 patient samples from The Cancer Genome Atlas 

and validated it using 1624 patient samples from five independent 

cohorts. We further identified drug candidates that 

can reverse the HCC disease gene expression, from a library 

consisting of over 1000 FDA-approved drugs. We identified 

niclosamide and other antihelminthics as potential drug candidates 

for the treatment of HCC, and evaluated niclosamide and 

niclosamide ethanolamine (NEN, a water soluble niclosamide 

salt), for their in vitro and in vivo anti-tumor efficacy against 

HCC cells and patient-derived HCC xenografts (PDHX). Results 

In silico analysis indicated niclosamide could reverse the gene 

expression of HCC patients regardless of their etiology and 

tumor grade (FDR


cedure. A novel surgical approach, ALPPS (Associating Liver 

Partition and Portal Vein Ligation for Staged Hepatectomy), 

which combines PVL with parenchymal transition, may be used 

for unresectable disease due to its ability to markedly accelerate 

liver regeneration via unknown mechanisms. We hypothesize 

that transection is sufficient to activate plasma proteins 

necessary for accelerated liver regeneration, and that portal 

vein ligation (PVL) is necessary to stimulate liver growth. Aim 

Using a novel mouse model, we aim at studying the molecular 

mechanisms underlying the accelerated liver growth associated 

with ALPPS. Methods Liver tissue was collected at early time 

points after ALPPS (30min, 1h, 4h, 8h, 12h) and analysed by 

RNA deep sequencing. Plasma samples were collected 30min 

after ALPPS Step 1 for proteomics analysis by protein enrichment, 

albumin depletion, and mass spectrometry. Each protein 

fraction is confirmed for activity by inducing accelerated regeneration 

upon injection into mice treated with PVL only. Results 

Injection of plasma, derived from mice, subjected to ALPPS, 

into PVL alone mice is sufficient to accelerate liver regeneration 

as observed following ALPPS surgery. Likewise, a comparable 

response is induced in PVL alone mice by plasma from 

mice subjected to transection only. Plasma proteins appear 

to chiefly account for the accelerated liver growth. Transcript 

analysis shows that differential gene expression occurs 4 hours 

post ALPPS. The accelerated liver growth is characterized by 

a hyperplastic reaction reflected in an increased number of 

hepatocytes entering the cell cycle as early as 8 hours after 

surgery. Conclusions ALPPS is conducive to accelerated liver 

regeneration through proteins circulating in the plasma as early 

as 30 minutes after the transection step. ALPPS-specific gene 

expression changes are observed in liver at early times after 

surgery and are consistent with the induction of regenerative 

pathways. Identification of plasma proteins stimulating liver 

growth and associated regenerative pathways may have a 

high therapeutic value in expanding the surgical cure of liver 

disease previously deemed unresectable. 

Disclosures: 

The following authors have nothing to disclose: Magda Langiewicz, Andrea 

Schlegel, Bostjan Humar, Rolf Graf, Pierre-Alain Clavien 

500 

Identifying novel therapeutic targets in HCC through 

an integrated transcriptomics and pharmacogenomics 

approach 

Bin Chen 1 , Li Ma 2 , Mei-Sze Chua 2 , Atul Butte 1 , Samuel K. So 2 ; 

1 Institute for Computational Health Sciences, UCSF, San Francisco, 

CA; 2 Asian Liver Center, Stanford University, Stanford, CA 

Background Large-scale gene expression profiling of disease 

systems perturbed by chemical agents in preclinical models 

allows the understanding of underlying cellular responses 

induced by the chemicals. Such information may guide the discovery 

of new therapeutic drugs and targets in specific disease 

systems. Using hepatocellular carcinoma (HCC) as our disease 

model, we hypothesized that genes whose expressions are specifically 

reversed by anti-cancer drugs may be potentially novel 

therapeutic targets in HCC. Methods and Materials We compiled 

data on IC 50 

s of anti-cancer drugs from ChEMBL (a chemical 

database of bioactive molecules with drug-like properties), 

and drug-induced gene expression signatures from LINCS (a 

database containing perturbation profiles of 1,000 genes in 

HCC cell lines) and from CMap (a database containing whole 

genome perturbation profiles in all cancer cell types). Additionally, 

we computed gene expression signatures of HCC and 

non-tumor liver samples from TCGA. We compared the HCC 

gene expression signature with the drug signatures, and computed 

the ability of each drug to reverse the HCC signature. 

Each drug was associated with a Reversing Gene Expression 

Score (RGES), and the correlation between the IC 50 

s and RGES 

of 36 drugs was examined. We further identified genes that 

are specifically reversed by anti-cancer drugs with IC 50 

s


tion in HCC cell lines and enhanced the nuclear translocation 

of TFEB-GFP and p62 clearance in all HCC cell lines. Western 

blot analysis showed impaired expression of lysosomal 

hydrolases cathepsin D in all HCC cell lines except HepG2 

cells. Using real-time RT-PCR assay, we found that mRNA levels 

of number of lysosomal genes (TFEB, cathepsin D) were significantly 

reduced in all HCC cell lines compared to primary 

human hepatocytes. Furthermore, we investigated whether 

autophagy inhibitor (hydroxychloroquine) or inducer (Torin 1) 

could be used as a therapeutic strategy for HCC. Interestingly, 

long-term culture of HCC cells with autophagy inhibitor leads 

to complete inhibition of HCC growth by MTT and cell colony 

assay. Conclusion: Our results show that increased expression 

of lysosomal efflux protein (p62) in HCC cell lines is due to 

defects in mTOR-TFEB signaling. Our results also show that the 

autophagy inhibitor hydroxychloroquine can be a potential 

agent to inhibit HCC growth. 

Disclosures: 

The following authors have nothing to disclose: Rajesh Panigrahi, Partha K. Chandra, 

Pauline Ferraris, Fatma Aboulnasr, Srinivas Chava, Tong Wu, Srikanta Dash 

502 

Chimeric hepatitis E virus-like particles as a tool of oral 

vaccination against hepatitis A virus 

Eun Byul Lee 1 , Jung-Hee Kim 1 , Jung Eun Choi 1 , Seung Kew 

Yoon 1,2 ; 1 The Catholic University Liver Research Center & WHO 

Collaborating Center of Viral Hepatitis, Seoul, Korea (the Republic 

of); 2 College of Medicine, The Catholic University of Korea, Seoul, 


Background: Virus-like particles (VLPs) has been shown the possibility 

of new tools for oral vaccination because VLPs can be 

protected against harsh environment of the human digestive 

tract. Recent studies have demonstrated that hepatitis E virus 

(HEV)-LPs can induce both mucosal and systemic immunity after 

oral administration. Currently, commercial vaccine against 

hepatitis A virus (HAV) are in the intravenous form, but those 

for oral administration are yet to be developed. In this study, 

we investigated whether HEV-LPs containing HAV antigens 

(HEV-LP-HAVag) could be useful delivery tool for oral vaccination 

against HAV. Methods: To produce HEV-LPs from mammalian 

cells, Huh7 cells were infected with the recombinant 

baculovirus containing CMV promoter derived-Nt-ORF2 gene 

(Bac-Nt-ORF2). Nt-ORF2 expression in Huh7 cells was confirmed 

by western blot analysis, and HEV-LPs produced from 

Huh7 cells were purified by sucrose gradient centrifugation. 

The morphology of purified HEV-LPs was observed by electron 

microscopy (EM). Expression vectors of HAV antigens (VP1, 

VP3, VP1-P2A)-derived from HM175 strain were constructed 

by insertion of gene encoding each HAV antigen into pcDNA3 

vector in frame. To establish HEV-LPs packing system carrying 

HAV antigen expression vectors, HEV-LPs were disassembled 

using biochemical buffer containing DTT and low concentration 

of CaCl 2, 

and then HAV antigen expression vectors were 

packed into the re-assembled HEV-LPs by increasing concentration 

of CaCl 2. 

Delivery of HAV antigen expression vectors and 

their expression were confirmed in Huh7 cells infected HEV- 

LP-HAVag by western blot analysis. Results: Nt-ORF2 was successfully 

expressed in Huh7 cells infected with Bac-Nt-ORF2. 

Moreover, Nt-ORF2 expressed in Huh7 cells formed HEV-LPs 

of 25 nm in diameter through their self-assembly property. Furthermore, 

HEV-LP-HAVags were efficiently build up by in vitro 

disassembly/reassembly systems and, their morphology was 

well preserved compared to that of empty HEV-LPs. By infection 

with HEV-LP-HAVags, HAV antigens expression vectors were 

delivered to Huh7 cells, and subsequently 3 kinds of HAV antigens 

were efficiently expressed in the cells. Conclusion: We 

established an antigen-encapsulation system with HEV-LPs and 

successfully delivered the antigen expression vectors using the 

system. This might be useful tool for development of vaccine as 

well as vesicles for gene therapy. 

Disclosures: 

The following authors have nothing to disclose: Eun Byul Lee, Jung-Hee Kim, Jung 

Eun Choi, Seung Kew Yoon 

503 

High-throughput sequencing to define the miRNA cargo 

of LSEC-derived, anti-viral exosomes 

Michael Kriss 1 , Michael Edwards 3 , Lucy Golden-Mason 1 , Colin 

Larson 4 , Katrina Diener 4 , Hugo R. Rosen 1,2 ; 1 Division of Gastroenterology 

& Hepatology, University of Colorado Anschutz Medical 

Campus, Aurora, CO; 2 Denver VA Medical Center, Denver, 

CO; 3 Division of Pulmonary Sciences and Critical Care Medicine, 

University of Colorado Anschutz Medical Campus, Aurora, CO; 

4 Genomics and Microarray Core, University of Colorado Anschutz 

Medical Campus, Aurora, CO 

Aim: To characterize the miRNA content of LSEC-derived exosomes 

to identify novel anti-viral mediators. Methods: Exosomes 

were isolated from supernatants of an immortalized LSEC cell 

line cultured for 48h ± rhIFNα (100ng/ml) or ± IFNλ1/2/3 

(100ng/ml each) using ExoQuick-TC (SBI). Total RNA was isolated 

using SeraMir RNA extraction kit (SBI). Small RNA library 

was generated using Illumina TruSeq Small RNA sample prep 

kit (1μg input RNA) and validated on Agilent Bioanalyzer. 

Sequencing was performed using Illmina HiSEQ 2500 Platform 

using V4 Chemistry Single Read 1x50. The sequenced reads 

were aligned with TopHat2 against both the human mirBASE 

and hg38 databases separately, and quantitated using Partek 

Genomics Suite v6.6. The average amount of reads produced 

from the miRNA and genome alignments were approximately 

3.3 and 9.2 million reads respectively per sample. Results: 

Exosomes derived from LSECs stimulated with either rhIFNα 

or IFNλ1/2/3 suppressed HCV replication in vitro and led 

to induction of ISGs in HCV-infected hepatocytes, consistent 

with prior reports. Electrophoresis of isolated RNA revealed 

absence of ribosomal RNA (18s and 28s) compared to RNA 

isolated from derivative cells. High throughput sequencing was 

performed with a base call accuracy of 99.96% (Phred Q score 

33.813±0.257). miRNA fold changes relative to mock were 

calculated based on read counts for all targets with greater 

than 100 total read counts. The top ten fold changes for each 

treatment group are included in table 1. miR-3168 was the 

only miRNA uniquely induced in the IFNλ-treatment group. 

miR-122 was present in all samples with modest induction in 

both IFNλ (1.31-fold) and IFNα (1.75-fold) treatment groups. 

Additional alignment with hg38 database revealed presence 

of miR-138 in only the IFNλ and IFNα treated groups. Conclusions: 

LSEC-derived exosomes differentially regulate miRNA in 

the setting of type I and type III interferons. This may contribute 

to the anti-viral effect of these exosomes and provide novel 

insights into therapeutic mechanisms moving forward.


Disclosures: 

The following authors have nothing to disclose: Michael Kriss, Michael Edwards, 

Lucy Golden-Mason, Colin Larson, Katrina Diener, Hugo R. Rosen 

504 

The Roles of Kras Isoforms in Liver Cancer 

Hyuk Moon 1,2 , Sook In Chung 1,2 , Kyung Joo Cho 1,2 , Hye Lim Ju 1 , 

Dayoung Kim 1 , Do Young Kim 1,3 , Sang Hoon Ahn 1,3 , Kwang- 

Hyub Han 1,3 , Weonsang S. Ro 1,3 ; 1 Liver Center, Yonsei University 

College of Medicine, Seoul, Korea (the Republic of); 2 Brain Korea 

21 PLUS Project for Medical Science College of Medicine, Yonsei 

University, Seoul, Korea (the Republic of); 3 Department of Internal 

Medicine, Yonsei University College of Medicine, Seoul, Korea 


Background/Aims: Activating mutation in Kras is frequently 

observed in human carcinogenesis. Alternative splicing of Kras 

transcripts gives rise to two isoforms, Kras4A and Kras4B. In 

this study, we compared the hepatocarcinogenic potentials of 

the KRAS splice variants carrying the same activating mutation. 

Methods: Transgenic liver cancer mouse models were 

developed using a hydrodynamic injection method and the 

Sleeping Beauty Transposon System. Transposons encoding 

Kras4AG12V or Kras4BG12V were mixed with transposons 

expressing short hairpin RNA downregulating p53 (shp53) and 

plasmids encoding the Sleeping Beauty transposases and then 

hydrodynamically delivered to the liver of 6-week-old C57BL/6 

mice. Livers were harvested and analyzed at 4 weeks post-hydrodynamic 

injection. Results: Co-expression of KRas4BG12V 

and shp53 resulted in massive abdominal enlargement within 

4 weeks after injection. Numerous nodular lesions emerged 

from the liver parenchyma and occupied most of the liver 

surface. In contrast, mice transfected with KRas4AG12V and 

shp53 had much smaller and fewer nodules in the liver. The 

ratio of liver weight/body weight in the Kras4BG12V group 

was significantly higher than that in KRas4AG12V group (p < 

0.001). Survival analysis also showed a significantly shorter 

life span for Kras4BG12V mice compared to Kras4AG12V 

mice (p


50% cell viability varies significantly among HCC cell lines. 

We have isolated HCC cell lines that were totally resistant 

to sorafenib and continuously grow in growth media supplemented 

with sorafenib. The mechanism of sorafenib resistance 

relates to the impaired uptake due to the reduced expression of 

organic cation transporter-1 and 3 (OCT1& OCT3). The uptake 

of doxorubicin is not altered in HCC cell lines. We found that 

OCT1 mRNA and protein expression is reduced in all HCC 

cell lines compared to primary human hepatocytes. Stable 

expression of full-length OCT1 in the resistant cell line induced 

cellular cytotoxicity to sorafenib and overcome the resistance. 

Conversely, we showed that inhibiting OCT1 expression by 

small molecule inhibitor (quinine hydrochloride) in a sensitive 

cell line reduced sorafenib uptake and induces resistance. 

The expression of OCT1 and OCT3 is significantly reduced 

in human hepatocellular carcinoma tissues as compared to the 

surrounding non-tumorous hepatocytes in the cirrhotic nodules. 

Conclusions: Our results indicate that there are intrinsic differences 

among HCC cell clones that affect sorafenib sensitivity. 

Expression of OCT1 and OCT3 down regulation in human 

samples is associated with sorafenib sensitivity. We propose 

that a detailed understanding of the sorafenib sensitivity and 

resistance mechanisms should allow for novel treatment options 

to improve sorafenib chemotherapy response in patients with 

liver cancer. 

Disclosures: 

The following authors have nothing to disclose: Srinivas Chava, Partha K. Chandra, 

Rajesh Panigrahi, Pauline Ferraris, Fatma Aboulnasr, James Liu, Jose J. 

Marin, Swan N. Thung, Tong Wu, Srikanta Dash 

507 

Stabilin-mediated endocytosis of antisense oligonucleotides 

for modulation of liver gene expression 

Colton M. Miller 1 , Aaron J. Donner 2 , Emma E. Blank 1 , Punit Seth 2 , 

Edward N. Harris 1 ; 1 Biochemistry, University of Nebraska, Lincoln, 

NE; 2 ISIS Pharmaceuticals, Inc., Carlsbad, CA 

Introduction: Antisense oligonucleotides (ASOs) are short 

(5000-7400 Da) chemically-modified oligonucleotides that 

bind to cellular RNAs and modulate their intermediary metabolism 

to produce a pharmacological effect. DNA-based ASOs 

down-regulate gene-expression by promoting RNase H mediated 

degradation of the targeted mRNA. There are currently 

over 40 ASOs in clinical development targeting genes that are 

involved with metabolic diseases, cancer and inherited genetic 

diseases. An important difference between natural DNA and 

the ASO is that the phosphodiester (PO) backbone of DNA has 

been partially or fully substituted with a phosphorothioate (PS) 

backbone. The replacement of the negatively charged oxygen 

for the negatively charged sulfur atom covalently bound to 

phosphorus renders the short polymer both high stability in 

biological fluids and a propensity to adhere to cell surfaces. 

Some ASOs are further modified by 2’-O-methoxyethyl (MOE) 

or 2’-O-Methyl RNA nucleotides to further enhance metabolic 

stability and RNA-binding affinity. Injected ASOs accumulate 

in the liver due to the scavenging function of the organ. Previous 

work has shown that the liver sinusoidal endothelial cells 

(SECs) and Kupffer cells (KC) internalize a much greater proportion 

of ASO than hepatocytes. We have discovered that the 

non-DNA binding Stabilin-2/HARE receptor specifically binds 

to and internalizes several classes of ASO as this scavenger 

receptor is enriched in these liver cells. Methods: In our system, 

we used stable cell lines expressing recombinant human Stabilin-1 

and -2 to assess internalization rates and wildtype CD-1 

mice to determine 125 I-ASOs in organ distribution and uptake 

in hepatic cells. Results: Cell lines expressing either Stabilin-1 

or Stabilin-2 or the parental (non-Stabilin) empty vector (EV) cell 

line were incubated with 125 I-ASO that was fully modified with 

PS at 0°C (binding) and 37°C (endocytosis) and determined 

that the amount of ASO bound did not differ, though the Stabilin-2 

cells internalized 3-fold more than the Stabilin-1 cells and 

7-fold more than the EV cells. Twenty percent of the retro-orbitally 

injected ASO accumulated in liver with an amount of 48%, 

30%, and 22% in SECs, KCs and hepatocytes, respectively. 

Despite progress in the use of therapeutic oligonucleotides, the 

pathways by which negatively charged ASOs enter cells and 

modulate gene expression remain poorly understood. Conclusion: 

In this context, our work provides fundamental insights 

into cell-surface proteins which mediate ASO uptake into cells 

and provide opportunities to leverage these findings to further 

enhance the therapeutic properties of oligonucleotide drugs. 

Disclosures: 

Aaron J. Donner - Employment: Isis Pharmaceuticals; Stock Shareholder: Isis 


Punit Seth - Employment: Isis Pharmaceuticals 

The following authors have nothing to disclose: Colton M. Miller, Emma E. Blank, 

Edward N. Harris 

508 

Shifting gut microbiota and bile acid profiles in retinoic 

acid-primed mice that exhibit accelerated liver regeneration 

Hui-Xin Liu, Ying Hu, Lili Sheng, Yu-Jui Yvonne Wan; Medical 

Pathology and Laboratory Medicine, UC DAVIS, Sacramento, CA 

Abstract Similar to bile acid (BA), all-trans retinoic acid (RA) 

facilitates 2/3 partial hepatectomy (PHx)-induced liver regeneration. 

RA can activate BA receptor farnesoid x receptor (FXR) to 

regulate BA signaling because retinoid x receptor α (RXRα) is 

a permissive partner of FXR. Our published data also showed 

that RA regulates hepatic lipid homeostasis by shifting hepatic 

BA profile. Since the gut microbiota plays a pivotal role in regulating 

BA homeostasis and BA composition, we examined the 

effect of RA in altering gut microbial and BA composition and 

their relationship with increased hepatocyte proliferation in 

response to liver resection. C57BL/6 mice were primed with a 

single dose of RA (25 μg/g) followed by PHx. RA-primed mice 

exhibited accelerated hepatocyte proliferation as indicated by 

higher number of Ki67-positive cells and earlier induction of 

cell cycle genes compared to untreated mice. Firmicutes and 

Bacteroidetes phyla, which affect the efficiency of host energy 

and are linked with adiposity in both mice and human, dominated 

the gut microbial community (>85%) in both control and 

RA-primed mice after PHx. RA reduced the ratio of Firmicutes 

to Bacteroidetes, which is associated with a lean phenotype. 

Consistently, RA-primed mice lacked transient lipid accumulation, 

an energy source for proliferating hepatocytes, normally 

found in regenerating mouse livers. This finding suggested that 

RA-primed mice might have increased lipid metabolism. In 

addition, RA altered BA signaling in the regenerating livers. 

RA activated the FXR-SHP-FGF15 pathway, reduced CYP7A1 

and CYP8B1 expression to inhibit BA synthesis, and increased 

BA transport through Oatp2, Asbt, and Ibabp induction in the 

ileum. These data suggested enhanced enterohepatic recirculation 

of BAs from RA-priming. RA treatment also shifted the 

BA profile by increasing the hydrophilic/hydrophobic ratio. 

The ratio of taurine-conjugated cholic acid (CA) to free CA 

and taurine-conjugated-β-murine CA to free β-murine CA were 

elevated in RA-treated regenerating livers suggesting increased 

solubility and lipid emulsification capability. Accordingly, fibroblast 

growth factor 21 (FGF21) and Sirtuin1 (SIRT1), master 

metabolic regulators, and their downstream targets AMPK and


ERK1/2 were more robustly activated in RA-primed regenerating 

livers than controls. Conclusion: Priming mice with RA 

resulted in a lean microbiota composition and hydrophilic bile 

acid profile, which are associated with facilitated metabolism 

and enhanced cell proliferation. 

Disclosures: 

The following authors have nothing to disclose: Hui-Xin Liu, Ying Hu, Lili Sheng, 

Yu-Jui Yvonne Wan 

509 

Pre-operative plasma glypican-3 levels detected by a 

novel ELISA system predict the risk of post-operative 

recurrence in patients with stage I hepatocellular carcinoma 

Kazuya Ofuji 1,2 , Keigo Saito 1 , Yasunari Nakamoto 2 , Tetsuya 

Nakatsura 1 ; 1 Division of Cancer Immunotherapy, Exploratory 

Oncology Research and Clinical Trial Center, National Cancer 

Center, Kashiwa, Japan; 2 Second Department of Internal Medicine, 

University of Fukui, Fukui, Japan 

Background: Prognosis of post-operative hepatocellular carcinoma 

(HCC) patients remains unsatisfactory due to the high 

risk of recurrence. Glypican-3 (GPC3) is specifically overexpressed 

in HCC and has been recognized as a novel prognostic 

factor after curative resection of HCC. Recent studies 

have shown the usefulness of GPC3 as a biomarker for the 

diagnosis of early HCC. However, the diagnostic value of 

GPC3 as a predictive marker for recurrence of postoperative 

early-stage HCC remains unclear. Here, this study was undertaken 

to evaluate the detection of plasma GPC3 using a novel 

sandwich enzyme-linked immunosorbent assay (ELISA) system 

in early-stage HCC patients after surgical resection. Methods: 

Plasma samples were collected from 25 patients with stage 

I HCC who underwent for surgical resection between 2008 

and 2010. The ELISA system for detection of plasma GPC3 

levels was established, by use of a newly-generated anti-GPC3 

mouse monoclonal capture antibody and a biotinylated detection 

antibody and recognized with streptavidin-conjugated 

horseradish peroxidase. GPC3 expression of surgical specimens 

was analyzed by immunohistochemical staining. Results: 

HCC recurrence was observed in 14 cases after surgical resection 

(51.8%) during follow-up period (median, 738 days). 

In the immunohistochemical analysis, GPC3 positive rate of 

resected specimen was 53% (13/23). There was a tendency 

towards a shorter recurrence-free survival (RFS) in GPC3 positive 

cases than in GPC3 negative cases (p 0.233). In the ELISA 

system, the mean ± standard deviation (SD) of plasma GPC3 

concentrations was 110.12 ± 37.70 ng/ml in control subjects, 

and cut-off value was determined to be 132 ng/ml (mean + 

1.5 SD). In stage I HCC, the sensitivity and specificity were 

40.7% and 92%, respectively. GPC3-positive rate at the time 

of recurrence was 64.3%. In the recurrence group, pre-operative 

plasma GPC3 levels were significantly higher than in the 

non-recurrence group (median, 191.7 ng/ml vs 29.7 ng/ml, 

p = 0.029). There was a tendency of shorter RFS in the pre-operative 

plasma GPC3-positive patients group compared with 

the negative group (median RFS, 769 days vs. not reached, 

p = 0.147). However, there was no significant difference in 

overall survival. The recurrence rate of patients with post-operative 

plasma GPC3 positive was significantly higher than 

GPC3 negative patients (p = 0.012). Conclusions: We have 

established a novel ELISA system for detecting plasma GPC3 

levels and evaluated the utility of GPC3 as a diagnostic marker 

of HCC. This study suggests pre-operative plasma GPC3 levels 

may help in identification of stage I HCC patients at high risk 

of post-operative recurrence. 

Disclosures: 

Tetsuya Nakatsura - Consulting: Ono Pharmaceutical co.Ltd.; Grant/Research 

Support: Ono Pharmaceutical co.Ltd., MEDINET co.Ltd., Sysmex co. Ltd. 

The following authors have nothing to disclose: Kazuya Ofuji, Keigo Saito, Yasunari 

Nakamoto 

510 

Evaluation of Glyoxylate and Hydroxyproline Metabolic 

Pathways Through the Use of Dicer-Substrate Small 

Interfering RNA 

Nicole Avitahl-Curtis 1 , Benjamin Holmes 1 , Luciano Apponi 2 , Rohan 

Diwanji 1 , Marita S. Larsson Cohen 2 , Chaitali Dutta 1 , Natalie W. 

Pursell 1 , Dongyu Chen 2 , Xue Shui 2 , Purva Pandya 2 , Utsav H. 

Saxena 2 , Martin Koser 2 , Abrams Marc 2 , Weimin Wang 2 , Hank 

Dudek 2 , Chengjung Lai 1 , Bob D. Brown 2 ; 1 Translational Biology, 

Dicerna Pharmaceuticals, Inc., Cambridge, MA; 2 Dicerna Pharmaceuticals, 

Inc., Cambridge, MA 

Primary Hyperoxaluria (PH) results from genetic mutations 

which affect glyoxylate metabolism, causing overproduction 

of oxalate. Three types of Primary Hyperoxaluria have been 

described to date, and the genetic lesions underlying disease 

have been identified. PH1, PH2 and PH3 are caused by mutations 

in the genes, AGXT, GRHPR and HOGA1, respectively, 

which encode the metabolic enzymes alanine: glyoxylate 

aminotransferase (AGT), glyoxylate reductase (GRHPR), and 

4-hydroxy-2-oxoglutarate adolase (HOGA). The roles of AGT 

and GRHPR in glyoxylate metabolism have been explored utilizing 

mouse strains harboring a germline disruption of the 

Agxt or Grhpr gene. Agxt- and Grhpr-deficient mice develop 

hyperoxaluria, consistent with the proposed roles of AGT and 

GRHPR in glyoxylate metabolism. Dicer-substrate siRNAs (DsiR- 

NAs) are potent and specific RNAi triggers that inhibit the 

expression of disease-relevant targets at the mRNA level, and 

are currently in clinical development. In this work, we describe 

the use of DsiRNAs to silence the expression of Agxt and Grhpr 

in mice to generate PH1 and PH2 animal models that phenotypically 

mimic Agxt- and Grhpr-deficient mice. DsiRNA-induced 

PH disease models were further explored for their utility 

to identify effective therapeutics. This approach was also used 

to investigate the dysregulation of the glyoxylate and hydroxyproline 

metabolic pathways occurring in each type of primary 

hyperoxaluria. Finally, we describe the utility of siRNA as an 

efficient and powerful method for generating models of liver 

disease in rodents and, potentially, other species. 

Disclosures: 

Nicole Avitahl-Curtis - Employment: Dicerna Pharmaceuticals, Inc. 

Benjamin Holmes - Employment: Dicerna Pharmaceuticals; Stock Shareholder: 

Dicerna Pharmaceuticals 

Luciano Apponi - Employment: Dicerna Pharmaceuticals 

Rohan Diwanji - Employment: Dicerna Pharmceuticals; Stock Shareholder: 

Dicerna Pharmceuticals 

Chaitali Dutta - Employment: Dicerna Pharmaceuticals 

Utsav H. Saxena - Employment: Dicerna Pharmaceuticals 

Martin Koser - Employment: Dicerna Pharmaceuticals 

Abrams Marc - Employment: Dicerna Pharmaceuticals 

Hank Dudek - Employment: Dicerna 

Chengjung Lai - Employment: Dicerna Pharmaceuticals 

Bob D. Brown - Employment: Dicerna Pharmaceuticals 

The following authors have nothing to disclose: Marita S. Larsson Cohen, Natalie 

W. Pursell, Dongyu Chen, Xue Shui, Purva Pandya, Weimin Wang


511 

Cell-Type-Specific Expression Profile of Repopulating 

Hepatocytes by Translating Ribosome Affinity Purification 

(TRAP) 

Kirk J. Wangensteen 1,2 , Amber W. Wang 2 , Monica Teta-Bissett 2 , 

Klaus H. Kaestner 2 ; 1 Gastroenterology, University of Pennsylvania, 

Philadelphia, PA; 2 Genetics, University of Pennsylvania, Philadelphia, 

PA 

Understanding the genetic regulation of liver regeneration 

could help in the development of new treatments for liver disease. 

Much of the research on liver regeneration has focused 

on the partial hepatectomy model in rodents; however, the lack 

of cell necrosis and immune response calls for the development 

of novel paradigms that recapitulate human liver diseases. Fah - 

/- 

mice, a model of hereditary tyrosinemia, are suitable for the 

study of liver repopulation after injury because repopulating 

hepatocytes can be genetically traced. We first injected Fah - 

/- 

mice with plasmids that co-express Fah and GFP, and found 

that we could isolate GFP-positive hepatocytes by fluorescence 

activated cell sorting, but met challenges in isolating pure populations 

of repopulating hepatocytes in the setting of injury. We 

then employed a different approach, the Translating Ribosome 

Affinity Purification (TRAP) system, to isolate mRNA specifically 

from repopulating hepatocytes. We injected Fah -/- mice with 

a plasmid that co-expresses Fah and a fusion of GFP and the 

ribosomal subunit L10a, and then induced liver injury for 1 or 

4 weeks. Liver tissue harvested from these mice was immediately 

lysed and polysomes were extracted. Anti-GFP antibodies 

were then used to affinity-purify the mRNAs bound to the GFP- 

L10a fusion protein expressed within repopulating hepatocytes 

(Figure). No RNA was obtained from wildtype control liver, 

as expected. We performed high throughput mRNA sequencing 

(RNA-seq) to identify differentially expressed genes during 

repopulation as compared to quiescent hepatocytes. Gene 

ontology analysis showed that the genes were enriched in metabolic 

pathways and in c-Myc-responsive genes. In conclusion, 

this represents the first analysis of the differential expression 

of repopulating hepatocytes in an injury model. Intriguingly, 

liver repopulation involves differential regulation of metabolic 

genes, likely to maintain metabolic homeostasis in the setting 

of widespread liver injury. 

Bioanalyzer tracings of affinity-purified RNA from mice treated 

with (Green and Red), or without (Blue) the TRAP vector, illustrating 

the specificity of the system. 

512 

In vivo distribution of exosome packaged extracellular 

miRNA-155 to the liver, hepatocytes and Kupffer cells 

and its functional effects 

Shashi Bala 1 , Timea Csak 1 , Fatemah Momen-Heravi 1 , Dora Lippai 

1 , Karen Kodys 1 , Donna Catalano 1 , Abhishek Satishchandran 1 , 

Victor Ambros 2 , Gyongyi Szabo 1 ; 1 Medicine, UMass Medical 

School, Worcester, MA; 2 Molecular Medicine, UMass Medical 

School, Worcester, MA 

Purpose: Circulating miRNAs are gaining increasing interest 

as new biomarkers of diseases and means of intercellular communication. 

Previously, we showed induction of miR-155 in the 

plasma and found that miR-155 was associated with extracellular 

vesicles (EVs). However, little is known about the half-life 

and biodistribution of EVs associated miRNAs in vivo. Here, we 

established an exosome-based miR-155 mimic delivery system 

to study the biodistribution and half-life of EV associated miR- 

155 using a miR-155 knock out (KO) mouse model. Methods: 

Female wild type (WT) (C57/BL6J) or miR-155 KO mice were 

used. Exosomes were isolated from murine B cells using filtration 

method and characterized by electron microscopy, Nanosight 

and Western blot analyses. Electroporation was used 

to introduce miR-155 mimic into exosomes. Purified miR-155 

or control mimic loaded exosomes were administered intravenously 

to miR-155 KO mice. Mice were perfused to eliminate 

blood contamination. The Mann-Whitney test was employed 

for statistical analysis. Results: Our results indicate that administration 

of exosomes loaded with miR-155 mimic resulted in a 

rapid accumulation of miR-155 in the plasma of recipient miR- 

155 KO mice with subsequent distribution in the liver, adipose 

tissue, lung, muscle and kidney (highest to lowest, respectively). 

At the cellular level, miR-155 was detected in hepatocytes and 

liver mononuclear cells of recipient miR-155 KO mice in vivo, 

suggesting successful uptake of exosomal miR-155 by various 

cell populations of the liver. Functionally, exosomes-mediated 

restoration of miR-155 in Kupffer cells isolated from miR-155 

KO mice resulted in the induction of pro-inflammatory cytokines 

(MIP2 and MCP1) after LPS challenge in an in vitro co-culture 

system. To mimic the half-life and distribution of miRNA naturally 

present in the plasma, we administered (iv) WT plasma 

(donor) to miR-155 KO (recipient) mice. WT plasma was generated 

from CpG+LPS treated WT mice and was enriched in miR- 

155. Our results indicate detection of donor miR-155 in plasma 

of recipient miR-155 KO mice and distribution to the liver and 

adipose tissue. Induction of pro-inflammatory cytokines was 

found in recipient miR-155 KO mice suggesting some biological 

activity. In summary, our results demonstrate tissue biodistribution 

and biologic function of EV-associated miR-155. 

Conclusion: Exosome packaged miR-155 is readily distributed 

to the liver in a cell–specific manner resulting in KC activation. 

These observations will aid in developing new exosome-mediated 

miRNA delivery and investigating biological function of 

other extracellular miRNAs in vivo. 

Disclosures: 

The following authors have nothing to disclose: Shashi Bala, Timea Csak, 

Fatemah Momen-Heravi, Dora Lippai, Karen Kodys, Donna Catalano, Abhishek 

Satishchandran, Victor Ambros, Gyongyi Szabo 

Disclosures: 

The following authors have nothing to disclose: Kirk J. Wangensteen, Amber W. 

Wang, Monica Teta-Bissett, Klaus H. Kaestner


513 

Integrated proteomics analyses identified a 

phosphoprotein relevant to apoptosis resistance in 

human hepatocellular carcinoma 

Hideaki Naoe 1 , Yohmei Hoshida 1 , Seiichiro Yokote 1 , Satomi 

Fujie 1 , Takehisa Watanabe 1 , Katsuya Nagaoka 1 , Taiji Yamazoe 

1 , Hiroko Setoyama 1 , Motohiko Tanaka 1 , Jiro Fujimoto 3 , Norie 

Araki 2 , Yutaka Sasaki 1 ; 1 gastroenterology and hepatology, Kumamoto 

University, Kumamoto, Japan; 2 Tumor Genetics and Biology, 

Kumamoto University, Kumamoto, Japan; 3 Surgery, Hyogo College 

of Medicine, Nishinomiya, Japan 

Aim: This study was aimed to identify the key molecules which 

account for apoptosis resistance of hepatocellular carcinoma 

(HCC) through integrated comprehensive analyses not only 

of gene and protein expression but also of post-translational 

modification which strictly regulate protein function and biology. 

Materials & Methods: Human HCC cells were stimulated 

with or without H 2 

O 2 

, an apoptosis-inducing stimulation. Protein 

and mRNA samples were extracted from these cells, and 

subjected to two-dimensional fluorescence differential gel electrophoresis 

(2-D DIGE) and cDNA microarray, respectively. 

After these analyses, we integrated obtained data to identify 

differentially expressed genes and proteins. Proteins that 

showed a significant change in phosphorylation after H 2 

O 2 

stimulation were further characterized by mass spectrometry. 

To block the phosphorylation of the target protein, we replaced 

a major phosphorylation site threonine by an alanine and stably 

expressed the wild-type protein and its TA mutant in HCC 

cells. MiRNA extracted in the same manner described above 

were subjected to miRNA array analysis. Obtained data were 

reprocessed for network analysis focused on the target protein. 

Finally, expression and phosphorylation of the protein was 

determined in surgically resected human HCCs and in adjacent 

liver tissues. Results: Broad range of around 30 proteins, 

including several classes of cytoskeletal proteins or molecular 

chaperons exhibited a significant phosphorylation change in 

response to apoptosis stimulation. Based on pathway analysis, 

we focused on one of the most prominently changed protein, 

nucleophosmin (NPM). Sensitivity to apoptosis stimulation varied 

among the cell lines, and decrease in NPM phosphorylation 

was more prominent in apoptosis-sensitive cell line than in 

apoptosis-refractory cell line. Knockdown of NPM using NPM 

specific siRNA in HCC cells enhanced cell death when treated 

with H 2 

O 2 

. Functional disruption of NPM phosphorylation in 

HCC cells reduced their apoptosis resistance. Network analysis 

indicated that miRNA X is associated with NPM phosphorylation 

via proteins such as PTEN and Cdk. Expression and 

phosphorylation of NPM were significantly enhanced in human 

HCC tissues compared to the adjacent liver tissues (n=23). 

Recurrence-free survival after surgical resection is significantly 

longer in HCC with lower expression of NPM phosphorylation 

group (n=14) than higher expression group (n=9). Conclusion: 

Our integrated proteomics strategy demonstrates that apoptosis 

resistance of human HCCs is, in part, accounted for by phosphorylation 

of NPM and downstream signaling, which may be 

a novel therapeutic target for human HCC. 

Disclosures: 

Yutaka Sasaki - Grant/Research Support: Chugai Pharmaceutical Co. JAPAN, 

MSD Co. JAPAN 

The following authors have nothing to disclose: Hideaki Naoe, Yohmei Hoshida, 

Seiichiro Yokote, Satomi Fujie, Takehisa Watanabe, Katsuya Nagaoka, Taiji 

Yamazoe, Hiroko Setoyama, Motohiko Tanaka, Jiro Fujimoto, Norie Araki 

514 

YAP Promotes Cyclin D1 Expression in Cholangiocytes 

and Hepatocytes 

Quy P. Nguyen, Ying Wan, Fanyin Meng, Haibo Bai; BaylorScott&White 

Hospital, Temple, TX 

Background: YAP is a critical regulator of bile duct specification, 

cholangiocyte and hepatocyte proliferation and survival. 

Elevated YAP activity stimulated bile duct development, cholangiocyte 

and hepatocyte proliferation and survival. In agreement, 

Yap deficiency results in bile duct paucity and reduced 

cholangiocyte/hepatocyte proliferation and liver survival. 

However, how YAP regulates these events is less understood. 

We aim to identify YAP’s downstream target in the liver. Methods: 

Immunohistochemistry of Cyclin D1 was performed in liver 

sections. Bile ducts and hepatocytes were isolated from H&E 

stained frozen liver sections with laser capture microscope. 

mRNA levels of Cyclin D1 were measured by real-time PCR. 

Results: In both developing livers and adult livers, Cyclin D1 is 

highly expressed in cholangiocytes. When knocking out Yap 

in the developing liver with Alb-Cre;Yap flox/flox mice, Cyclin 

D1-positive biliary cells on ductal plates are significantly less 

than WT controls. When increasing YAP activity through ablating 

upstream negative regulator NF2 with Alb-Cre;Nf2 flox/flox 

mice, the number of Cyclin D1-positive cells on ductal plates 

increases dramatically. In adult liver, when inducing YAP 

overexpression in hepatocytes of Yap transgenic mice, there 

is a compensatory loss of YAP expression in cholangiocytes. 

Hepatocyte YAP overexpression induces Cyclin D1 expression 

in hepatocytes. The loss of YAP expression in cholangiocytes 

results in corresponding loss of Cyclin D1 expression in cholangiocytes. 

mRNA analysis of bile ducts and hepatocytes isolated 

from Yap Tg mice with laser capture microscope revealed that 

YAP did not regulate Cyclin D1 mRNA transcription. Conclusions: 

Because Cyclin D1 is known to promote proliferation 

and survival, YAP may regulate hepatocyte and cholangiocyte 

proliferation and survival through promoting Cyclin D1 activity. 

Cyclin D1 inhibitor represent another treatment strategy for 

YAP overexpression liver cancer. 

Disclosures: 

The following authors have nothing to disclose: Quy P. Nguyen, Ying Wan, 

Fanyin Meng, Haibo Bai 

515 

Therapeutic Potential in Toxic Liver Injury of Exosomes 

Isolated from Healthy or Diseased Donors Depends on 

their Content with Capacity to Beneficially Regulate Cellular 

Gene Expression 

Yogeshwar Sharma 1 , Tatyana Tchaikovskaya 1 , Preeti Viswanathan 

2 , Charles E. Rogler 1,3 , David B. Rhee 4 , Pilib Ó Broin 4 , Wilber 

Quispe 4 , Alexander Y. Maslov 4 , Aaron Golden 4 , Sanjeev 

Gupta 1,5 ; 1 Medicine, Albert Einstein College of Medicine, Bronx, 

NY; 2 Division of Pediatric Gastroenterology, Children’s Hospital 

at Montefiore Medical Center, Bronx, NY; 3 Department of Immunology 

& Microbiology, Albert Einstein College of Medicine and 

Montefiore Medical Center, Bronx, NY; 4 Department of Computational 

Sciences, Albert Einstein College of Medicine and Montefiore 

Medical Center, Bronx, NY; 5 Department of Pathology, 

Albert Einstein College of Medicine and Montefiore Medical Center, 

Bronx, NY 

The biological role and therapeutic potential of exosomes 

recently gathered much interest. Exosomes contain a variety 

of materials, including cellular proteins, mRNA and microRNA 

species, lipids, small biomolecules, etc. When isolated exosomes 

are injected into subjects, these are efficiently targeted to


liver with incorporation in hepatocytes. To elucidate the ability 

of exosomes to alter pathophysiological events and processes 

in recipients and to determine donor-dependent potential of 

exosomes, we studied healthy C57BL/6 mice and mice given 

single LD50 dose of acetaminophen (APAP) with liver injury 

characterized by abnormal liver tests and hepatic necrosis. 

Exosomes were isolated from mouse sera and medium from 

hepatocytes in cell culture. Also, exosomes were isolated from 

human sera. Administration of exosomes with reporters showed 

these were efficiently incorporated in cultured hepatocytes and 

intact mouse liver. In mouse or human hepatocyte cell culture 

assays with MTT for APAP cytotoxicity, exosomes from healthy 

mouse or human sera and culture medium from healthy hepatocytes, 

but not APAP-treated cells or mice, were cytoprotective. 

Administration of healthy exosomes after APAP in C57BL/6 

mice decreased mortality significantly, p


of HSC. We are now carrying out the molecular analyses of 

CYGB promoter activation stimulated by phosphorylated JNK. 

Disclosures: 



Gilead, BMS 



The following authors have nothing to disclose: Misako Sato, Matsubara, Tsutomu 

Matsubara, Atsuko Daikoku, Krista Rombouts, Kazuo Ikeda 

518 

Impaired skeletal muscle mitochondrial respiration 

during hyperammonemia of cirrhosis contributes to sarcopenia 

Julie H. Rennison 2 , Avinash Kumar 2 , Gangarao Davuluri 2 , Allawy 

Allawy 2,4 , Rafaella Nascimento e Silva 2 , Dharmvir Singh 2 , David 

R. Van Wagoner 5 , Hoppel Charles 3 , Srinivasan Dasarathy 1 ; 

1 Department Of Gastroenterology and Hepatology, Cleveland 

Clinic, Cleveland, OH; 2 Pathobiology, Cleveland Clinic, Cleveland, 

OH; 3 Pharmacology and Medicine, Case Western Reserve 

University, Cleveland, OH; 4 Department Of Internal Medicne, 

Cleveland Clinic, Cleveland, OH; 5 Molecular Cardiology, Cleveland 

Clinic, Cleveland, OH 

Hyperammonemia is a consistent abnormality in cirrhosis. 

Skeletal muscle metabolic disposal of ammonia in cirrhosis 

has been shown to contribute to sarcopenia or loss of skeletal 

muscle mass. Ammonia disposal occurs in muscle mitochondria 

and increased fragmentation and impaired ATP content occur 

in the muscle during hyperammonemia of cirrhosis. We tested 

the hypothesis that mitochondrial oxidation is decreased during 

hyperammonemia and identified the specific sites on the electron 

transport chain complexes (ETCC) that contribute to the 

impaired mitochondrial respiration. We evaluated the impact 

of hyperammonemia (AmAc) on mitochondrial respiration 

in differentiated C2C12 murine myotubes by high-resolution 

respirometry in the basal state in intact cells and in response 

to specific substrates for the ETCC (pyruvate for complex I, 

succinate for complex II, TMPD for complex III) and inhibitors 

of each of the ETCC (rotenone for complex I, oligomycin for 

ATP synthetase, FCCP for uncoupling oxidative phosphorylation, 

antimycin A for Complex III). Control and AmAc treated 

cells were were assessed in separate chambers simultaneously. 

Hyperammonemia decreased basal respiration in intact cells 

by ~35% (Control, 67.0±14.1; 24hr AmAc 42.3±7.9 pmol 

O 2 

s -1 10 -6 ) (p


way, thereby also significantly decreasing the expression of the 

FXR-inducible target genes OSTα/β and OATP1B3. We show 

a new miRNA-dependent mechanism of FXR regulation, which 

could affect the expression of FXR target genes as shown here 

for colon and liver carcinoma derived cell lines and tissue. 

Disclosures: 

The following authors have nothing to disclose: Regina Krattinger, Jessica 

Mwinyi, Gerd A. Kullak-Ublick, Helgi B. Schiöth, Adrian Boström 

520 

Chenodeoxycholic acid induced changes in the expression 

of microRNAs and genes involved in lipid, bile acid 

and drug metabolism in human hepatocytes 

Gerd A. Kullak-Ublick 2 , Regina Krattinger 2 , Serene M. Lee 3 , 

Wolfgang E. Thasler 3 , Jessica Mwinyi 1,2 ; 1 Division of Functional 

Pharmacology, Department of Neuroscience, Uppsala University, 

Uppsala, Sweden; 2 Department of Clinical Pharmacology and Toxicology, 

University Hospital Zurich, Zurich, Switzerland; 3 Department 

of General, Visceral, Transplantation, Vascular and Thoracic 

Surgery, Grosshadern Hospital, Ludwig Maximilians University, 

Munich, Munich, Germany 

Background: Bile acids (BAs) are important for the absorption 

of dietary lipids, lipid soluble vitamins and xenobiotics. As 

ligands for different transcription factors, e.g. the farnesoid X 

receptor, they can act as gastrointestinal signaling hormones 

that influence lipid, glucose, and energy homeostasis. Both 

hepatic disease states as well as drugs can lead to cholestasis, 

which is characterized by elevated intrahepatic BA levels. 

Here, we studied to which extent BAs modulate the mRNAome 

and miRNAome in human hepatocytes. Methods: Five batches 

of primary human hepatocytes were treated with 50 μmol/L 

chenodeoxycholic acid (CDCA) for 24 or 48 hours. Total RNA 

was extracted, size fractionated and subjected to Next Generation 

Sequencing to generate comprehensive mRNA and 

miRNA profiles. Results: Expression of 738 mRNAs and 52 

miRNAs was significantly decreased, whereas 1566 mRNAs 

and 29 miRNAs were significantly increased in CDCA treated 

hepatocytes. Several mRNAs from within gene clusters that 

control both BA and lipid homeostasis (FGF(R), APO and FABP 

family members, HMGCS2) and drug metabolism (CYP, UGT 

and SULT family members) showed a modulated expression in 

response to CDCA. Furthermore, CDCA was able to modulate 

the expression of several microRNAs such as e.g. miR-149, 

-505, -885 and -452. Of note, miR-34a showed an inverse 

expression pattern in relation to a distinct cluster of mRNAs 

consisting of ABCG5 and ABCG8, SLCO1B1, SLC22A7 

and FGF19. Conclusions: CDCA alters intracellular levels of 

regulatory miRNAs such as miR-34a, thereby influencing the 

expression of genes that are regulated by these miRNAs. Gene 

clusters thus controlled by CDCA are involved in bile acid 

homeostasis. 

Disclosures: 

The following authors have nothing to disclose: Gerd A. Kullak-Ublick, Regina 

Krattinger, Serene M. Lee, Wolfgang E. Thasler, Jessica Mwinyi 

521 

A Regional Multidisciplinary Liver Tumor Board 

improves Access to Hepatocellular Carcinoma Treatment 

for Patients Geographically Distant from Tertiary Medical 

Center 

E. M. Egert 1 , Rena D. Johnson 2 , Micah M. Watts 3 , Jordan Booty 3 , 

Nevena Damjanov 3 , Rosemary Mohr 3 , Jenifer Griffiths 4 , Major 

Lee 3 , Haroon A. Shaikh 4 , Richard C. Hawley 4 , Martha S. Ghosh 5 , 

Nicole S. Mungiole 3 , David E. Kaplan 3 ; 1 Coatesville VA Medical 

Center, Cochranville, PA; 2 Wilmington VA Medical Center, Wilmington, 

DE; 3 Philadelphia VA Medical Center, Philadelphia, PA; 

4 Lebanon VA Medical Center, Lebanon, PA; 5 Wilkes-Barre VA 

Medical Center, Wilkes-Barre, PA 

Introduction: Optimal care for hepatocellular carcinoma (HCC) 

requires multiple disciplines with expertise not always available 

at non-tertiary medical centers. Distance from tertiary centers 

has previously been shown to negatively impact access to liver-related 

services such as transplantation in the VA (Goldberg 

DS, et al. JAMA 2014). The goal of the VISN4 HCC Initiative 

was to improve access to HCC care for patients diagnosed 

at distant hospitals. Methods: A regional liver cancer team 

was established in eastern Pennsylvania and Delaware with 

clinicians, radiologists and coordinators from 4 spokes centers 

(Coatesville, Lebanon, Wilkes-Barre and Wilmington) and one 

tertiary hub (Philadelphia). Telehealth infrastructure was created 

to foster face-to-face participation of referring providers 

with the central multidisciplinary liver tumor board (LTB) for 

weekly meetings starting July 2014. Telehealth infrastructure 

was also used clinically to facilitate patient-to-specialist consultation 

from remote sites. To assess the impact of this program, 

we collected the date of first radiologic or serologic (abnormal 

AFP) abnormality, the first cross-sectional contrast enhanced 

imaging (CT/MRI), the first consultation order to the tertiary 

center, the first cancer plan, and the first cancer-related intervention 

for patients diagnosed with HCC in all 5 centers in 

the 12 months preceding and 10 months following the first 

meeting of the LTB. Date differences for the pre- and post-initiation 

time frames were compared using Wilcoxon Rank Sum 

tests stratified by Spoke/Hub status. Results: 124 patients 

with HCC were diagnosed: 61 pre-LTB (spokes 37, hub 28) 

and 63 post-LTB (spokes 26, hub 37). Median age was 64; 

55% were white, 44% were black, and 1% other. The number 

of cases diagnosed by liver biopsy as opposed to imaging 

declined from 15/61 (24%) to 4/63 (6%) (p=0.006). Time 

between the first abnormality and evaluation by a specialist/ 

LTB declined from 44d (18-75) to 21d (11-35) (p=0.0018) 

primarily for cases at spokes (55d to 23d, p=0.0004). For 

patients diagnosed at a spoke facility, time from first abnormality 

to first treatment declined from 79d (62-124) to 54d (34-83) 

(p=0.025). The number of resections performed increased from 

0 pre-LTB to 6 post-LTB (p=0.015).Conclusion: The creation of 

a LCTB improved access to HCC care, reduced unnecessary 

biopsies, shortened time from identification to treatment primarily 

due to improvement from spokes sites. These data mimic 

HCC care access improvements achieved with a similar program 

in VISN3 suggesting that this model warrants exploration 

for further implementation with in the VA system and possibly 

in private settings. 

Disclosures: 

Nevena Damjanov - Grant/Research Support: ArQule; Speaking and Teaching: 

SIRTex, Bayer, Amgen 

David E. Kaplan - Grant/Research Support: Bayer Healthcare Inc, Gilead, Inovio 


The following authors have nothing to disclose: E. M. Egert, Rena D. Johnson, 

Micah M. Watts, Jordan Booty, Rosemary Mohr, Jenifer Griffiths, Major Lee, 

Haroon A. Shaikh, Richard C. Hawley, Martha S. Ghosh, Nicole S. Mungiole


522 

Diminished Health-Related Quality of Life is an Independent 

Predictor of Hospitalization in Patients with 

Cirrhosis 

Sujit V. Janardhan 1 , Jason Morris 1 , David G. Beiser 2 , Nancy 

Reau 1 ; 1 Medicine, Section of Gastroenterology, Hepatology and 

Nutrition, Center for Liver Diseases, University of Chicago Medicine, 

Chicago, IL; 2 Medicine, Section of Emergency Medicine, 

University of Chicago Medicine, Chicago, IL 

INTRODUCTION: Decreased health-related quality of life (QOL) 

has been shown to be predictive of adverse clinical outcomes 

in many disease states. Patients with cirrhosis have diminished 

QOL that worsens with disease progression. However, it is not 

known whether diminished QOL is an independent predictor 

of clinical outcomes, such as hospitalization. HYPOTHESIS: We 

hypothesized that decreased QOL is an independent predictor 

of hospitalization in patients with cirrhosis. METHODS: We performed 

a retrospective cohort analysis of patients with cirrhosis 

who were enrolled in the chronic liver disease (CLD) patient 

database at the University of Chicago between 11/2012 and 

11/2014. QOL scores (as measured by patient responses to 

the Chronic Liver Disease Questionnaire) and clinical data (e.g. 

compensated vs. decompensated cirrhosis) were collected at 

the time of enrollment into the database. Patients were assessed 

for the number and length of hospitalizations occurring from 

the time of enrollment until the end of available follow up. QOL 

scores were analyzed for both overall score and QOL quartile 

(first quartile = highest 25% of HR-QOL scores). Univariate 

(ANOVA) and multivariate (linear regression) analysis was 

performed to find independent predictors of hospitalization. 

RESULTS: 100 patients with cirrhosis were identified from a 

cohort of 579 patients enrolled in the CLD database including 

67 patients with compensated cirrhosis (Childs-Turcotte-Pugh 

(CTP) class A) and 33 patients with decompensated cirrhosis 

(CTP B = 31, 2 CTP C = 2). The average length of follow-up 

was 19.61 +/- 8.82 months. 40 patients had at least one hospitalization 

during the follow-up period, with an average length 

of stay of 8.95 +/- 8.66 days (range 1-37 days). In univariate 

analysis, the average number of days spent hospitalized varied 

significantly between patients with compensated cirrhosis vs. 

decompensated cirrhosis (2.07 +/- 3.93 vs. 6.64 +/- 10.26 

days, respectively; p


524 

Did improvement of access to healthcare change the 

clinical features and long-term outcomes of hepatocellular 

carcinoma in a suburban area in Japan? 

Teru Kumagi 2,1 , Takahide Uehara 2 , Masaki Ohmoto 2 , Atsushi 

Hiraoka 2 , Miyake Teruki 2,1 , Kazuhiro Tange 2 , Akiko Toshimori 2 , 

Norio Horiike 2 , Morikazu Onji 2 ; 1 Gastroenterology and Metabology, 

Ehime University Graduate School of Medicine, To-on, Japan; 

2 Saiseikai Imabari Hospital, Imabari, Japan 

Background and aim: Access to healthcare is an important 

factor that may affect the management of any disease. The 

aim of this study is to identify the change of clinical features at 

diagnosis and long-term outcomes of HCC, focusing on access 

to healthcare in a suburban area in Japan. Methods: Chart 

review was conducted for patients with HCC diagnosed at 

a single center. Collected data (1983–2011) was analyzed 

including the following data: demographics (age at diagnosis, 

sex, residential postcode), Child-Pugh (CP) score, clinical stage 

of HCC (TNM classification) at diagnosis, initial treatments 

(resection, ablation, TACE, palliative care) and outcomes. 

Patients were divided into 2 groups according to their residential 

postcodes for mainland and island, in which they needed 

to take ships to visit our hospital until the bridges opened in 

Apr 1999. Thus the era was divided into 2 groups based 

on the changes in the access pattern (former era, before Apr 

1999; latter era, after Apr 1999). Local ethic board approved 

the study but written consent form was waived due to the retrospective 

manner. Results: Data analyses were conducted in 

371 HCC patients (male 78% and 21% residing on island). 

Mean age was 64.4+/-9.4. Distribution of CP scores was as 

follows: A, 67%; B, 20%; and C, 13%. Distribution of clinical 

stages of HCC was as follows: I, 14%; II, 35%; III, 31%; IVa, 

18%; and IVb, 1.4%. Patients in the latter era (n=108) were 

diagnosed at earlier stages (Stages I+II) than that in the former 

era (n=263) (P=0.046). During the median observation period 

of 814 days, the latter era had a significantly longer survival 

rate than that of the former era (P


alcohol-attributable liver disease in USA is unknown. Therefore, 

the aim of this study is to investigate the epidemiological 

trends of alcohol liver disease in USA. Methods: The UNOS 

liver transplant registry and National Vital Statistics Systems 

(NVSS) was assessed for individuals with alcohol liver disease 

(ALD). Using the UNOS database, we identified all adult (> 

18 years) candidates during the 10-year study period (2004 

– 2013) who were (i) added to liver transplant waiting list 

and (ii) candidates who received liver transplant (LT). Using 

the NVSS database, we identified all adults (> 18 years) with 

a cause of death attributed to alcohol liver disease using the 

ICD code (K70). Data extracted from both (UNOS and NVSS) 

databases included patient demographics (i.e. age, gender 

and ethnicity), and from the UNOS database (MELD score and 

UNOS regions). The age groups in the UNOS database are 

categorized as (i) 18 – 34 years (ii) 35 – 49 years (iii) 50 – 64 

years and (iv) 65 + years and in the NVSS database (i) 18 – 

24 years (ii) 25 – 44 years (iii) 45 – 54 (iii) 55 – 64 years 

and (iv) 65 + years. Results: During the study period 15,608 

candidates with ALD were added to the waiting list (WL) and 

6,502 received liver transplant. In the NVSS mortality dataset, 

there were 150,613 with a primary cause of death attributed 

to ALD during the study period. In the latter dataset, among all 

individuals with a diagnosis of cirrhosis (ALD and non-ALD), 

there was a significant increase in mortality for the proportion 

with ALD from 46% (2004) to 50% (2013). However, age-specific 

analysis showed that the increase in mortality rate was 

only significant in the age group < 55 years. Regarding transplantation 

trends there was an increase in both the addition of 

candidates with ALD to the WL and increase in liver transplant 

for candidates with ALD, but only in those in the age group 

35 – 49 years. In this age group, there was an increase in WL 

from 14.2% (2004) to 28.4% (2013) and increase in LT from 

11% (2004) to 25.3% (2013). There was no change in either 

WLR or LT in the other age groups. Conclusion: The burden of 

ALD in USA in the last decade is increasing. ALD is becoming 

a major public health crisis in the younger age group, and it is 

now the most common indication for both liver transplant and 

wait list addition in the age group 35 -49 years. 

Disclosures: 

Edson S. Franco - Grant/Research Support: Biospheres Medical 

Angel Alsina - Speaking and Teaching: Bayer, Novartis 

The following authors have nothing to disclose: Nyingi M. Kemmer, Thure Caire 

527 

Evaluation of DAA (direct-acting antiviral) access across 

the US: the interplay between Hepatitis C (HCV) patient 

and insurance type 

Jason Katz, Samantha Fernando, Antoinette Wilson, James 

Deemer, Marijke Frantsen; Ipsos Healthcare, New York, NY 

BACKGROUND In the last 18 months, highly effective DAAs 

have become available for HCV treatment, but the cost of these 

drugs has made access challenging. In this paper we examine 

treatment decision-making across payers by patient type. 

METHOD Ipsos Healthcare’s HCV Therapy Monitor, running 

since 2005 in the USA, reports on >150 physicians per quarter, 

previously annually, across the USA. Physicians provide 

patient demographic, disease and treatment data on treated 

and untreated HCV patients seen within each study period. 

Here, new DAAs include all sofosbuvir- and dasabuvir-containing 

regimens. RESULTS Following the launch of sofosbuvir 

in December ‘13, HCV treatment rates increased from 12% 

to 23% from Q1’14 to Q1‘15. During Q1‘15, rates of treatment 

with new DAAs differed across insurance types; 25% 

of all privately insured patients were on new DAA regimens, 

versus 21% and 16% of Medicare and Medicaid patients, 

respectively. All untreated patients who had payer restrictions 

as a treatment barrier, regardless of payer type, rose from 

5% to 22% from Q1’14 to Q1’15. In Q1’15, physicians’ top 

barriers to prescribing new DAAs were restricted to certain 

patient type (24%) and insurance coverage (19%). Publically 

insured patients are more likely to have advanced fibrosis 

(F3-F4) (46%) than privately insured patients (29%). Amongst 

more advanced patients across all payer types, F3 patients 

rather than F4 patients are most likely to be treated with a 

new DAA; this is also observed throughout ‘14. This trend is 

more pronounced at the Medicaid group where half of currently 

untreated F4 patients are substance abusers and 92% 

suffer from another condition. Of the whole Q1’15 untreated 

population, 33% are substance abusers and 77% suffer from 

another concomitant disorder. The population treated with a 

new DAA includes 16% substance abusers and 66% suffering 

from comorbidity. CONCLUSION Treatment of HCV patients 

with a new DAA is heavily influenced by fibrosis, payer type, 

comorbidities, and substance abuse status. Publically insured 

patients are more likely to have advanced fibrosis than the 

privately insured but the latter are more likely to be on a new 

DAA. Untreated patients are more likely to be substance abusers 

and suffer from comorbidities. These results reflect the adjudications 

made by US payers, many deciding to treat patients 

with advanced fibrosis who are free of conditions that could 

mitigate treatment outcomes. This also highlights that a large 

proportion of the population suffering from advanced fibrosis 

currently face access barriers to new DAAs because of their 

patient profile and insurance coverage. 

Disclosures: 

Jason Katz - Consulting: Abbvie, Gilead, Bristol Myers Squibb 

The following authors have nothing to disclose: Samantha Fernando, Antoinette 

Wilson, James Deemer, Marijke Frantsen 

528 

Geriatrics and cirrhosis: changing epidemiology of 

chronic liver disease among the elderly, 2004-2013 

Michael Hagan, Maria A. Kouznetsova, Sumeet K. Asrani; Baylor 

University Medical Center, Dallas, TX 

Introduction Chronic liver disease (CLD) morbidity has 

increased over the last decade, with a marked increase in 

admissions among the elderly. Methods: We examined all CLD 

related encounters (2004-13) in the elderly (>65 years) in the 

Baylor Health Care System serving Dallas-Fort Worth in rural, 

urban and community settings (8 hospitals, catchment 7 million, 

>130,000 annual admissions). Results: There were 26,816 

CLD related admissions (2004-2013); 23% of the admissions 

were in the elderly. Elderly patients had a higher prevalence of 

NAFLD/cryptogenic cirrhosis (44%) vs. 31% (45-54) and 41% 

(55-64) and primary liver malignancy (6.6%) vs. 3.3% (45-54) 

and 4.5% (55-64). Comorbidities were high: CAD (25%), CHF 

(22%), COPD (13%), DM (44%). Infection was higher in the 

elderly (33%) vs. 26% (45-54) and 25% (55-64). Disposition 

to home was lowest (51%) and inpatient mortality/hospice was 

highest in the elderly (20%) vs. 11% (45-54) and 14% (55-64). 

From 2004-05 to 2012-13, the largest increase in admissions 

were ages 55-64 (126% increase) and 65+ (83% increase). 

In the elderly, admissions increased three fold (5.5 to 14.4%). 

NAFLD/cryptogenic liver disease increased (32% to 50%) and 

viral hepatitis decreased (17% to 9.5%). Comorbid conditions 

increased (p


26% to 19%. Paracentesis and thoracentesis increased, 12% 

to 20%. Conclusion: Elderly subjects with CLD present with 

more comorbidities, more complicated liver disease, higher 

prevalence of NAFLD rather than viral hepatitis, higher procedure 

utilization and substantial inpatient mortality. The epidemiology 

of CLD in the elderly is changing and warrants chronic 

disease management models to mitigate the burden expected 

to be borne by Medicare. 

Disclosures: 

The following authors have nothing to disclose: Michael Hagan, Maria A. Kouznetsova, 

Sumeet K. Asrani 

529 

Communication, sequence and system modeling of GI/ 

Hepatology outpatient and inpatient work flows for targeting 

clinical decision support tools 

Anne Miller 3,4 , Jejo D. Koola 3,4 , Michael E. Matheny 3,4 , Julie 

Ducom 1 , Jason M. Slagle 4 , Erik J. Groessl 1,5 , Sterling M. Dubin 1,2 , 

Freneka F. Minter 3,4 , Jennifer H. Garvin 6,7 , Matthew B. Weinger 4 , 

Samuel B. Ho 1,5 ; 1 VA San Diego Healthcare System, San Diego, 

CA; 2 Medicine, University of California, San Diego, San Diego, 

CA; 3 VA Tennessee Valley Healthcare System, Nashville, TN; 

4 Vanderbilt University, Nashville, TN; 5 University of California, 

San Diego, San Diego, CA; 6 VA Salt Lake City Healthcare System, 

Salt Lake City, UT; 7 University of Utah, Salt Lake City, UT 

Objectives: Increasing numbers of cirrhotic patients have high 

re-admission and mortality rates. Interventions that improve 

care coordination with well-targeted clinical decision support 

(CDS) can improve outcomes. We analyzed work processes 

in outpatient and inpatient settings to identify opportunities for 

CDS placement. Methods: Ethnographic observations, guided 

by user-centered design principles, occurred at two VA medical 

centers. Within 24 hours, pairs of experienced observers met 

to interpret their notes. The debriefings elucidated the structure 

and meaning of work as practiced resulting in: 1) Affinity diagrams 

to aggregate interpreted themes, insights & intervention 

design ideas; 2) Communication models to elucidate information 

flows across roles; 3) Sequence models to define work 

in goal-oriented terms, and 4) Artifact analyses to elucidate 

cognitive & problem solving structures. Results: At Site A, 26 

physicians, 3 nurses and 1 clerk were observed in: 1) gastrointestinal 

(GI) outpatient clinics (31%); 2) inpatient medical 

rounds (69%). At Site B, 14 physicians were observed in: 1) 

GI outpatient (14%) & Primary Care clinics (29%); 2) Emergency 

Department (21%); 3) inpatient medical rounds (29%) 

and 4) inpatient GI consult rounds (7%). 2/3 of participants 

were trainees & 1/3 were attending physicians. We categorized 

168 and 147 notes, respectively, during debriefings 

into 3 major themes each with several sub-themes. Team and 

Care Coordination addressed work allocation among local 

and distributed providers. Alerting and Reviewing addressed 

information access and retrieval so as to confer meaning to 

patients’ situations. Executing patients’ plans-of-care addressed 

the implementation of clinical goals & tasks. We identified 

resident physicians as the primary vertical and horizontal communication 

and coordination hubs. Using artifact analyses we 

found that residents’ paper patient printouts most effectively 

supported their clinical work. Using sequence analysis we identified 

patient assessment & planning as critical resident tasks 

to be targeted for CDS interventions (e.g., automated dashboards, 

data visualization, diagnostic and therapeutic checklists). 

Conclusions: The success of quality improvement depends 

on how well interventions are integrated into routine work. 

This study identified 4 intervention design goals to enhance 

cirrhosis care: 1) interventions should assume distributed work 

environs; 2) target residents; 3) integrate CDS with clinical 

assessment & planning processes rather than global point of 

order entry; and 4) be provided in both electronic & hardcopy 

forms. The design/integration of specific CDS tools is ongoing. 

Disclosures: 

Samuel B. Ho - Grant/Research Support: Genentech, Gilead; Speaking and 


The following authors have nothing to disclose: Anne Miller, Jejo D. Koola, 

Michael E. Matheny, Julie Ducom, Jason M. Slagle, Erik J. Groessl, Sterling M. 

Dubin, Freneka F. Minter, Jennifer H. Garvin, Matthew B. Weinger 

530 

Reduction of 30 day Readmission Rate in Patients with 

End Stage Liver Disease 

Simona Rossi 1 , Manisha Verma 1 , Catherine Reynolds 2 , Dee Morrison 

2 , June Smith 2 , Cindy Mcglone 1 , Eyob L. Feyssa 1 , Victor J. 

Navarro 1 ; 1 Medicine, Albert Einstein Medical Center Philadelphia, 

Philadelphia, PA; 2 Nursing, Albert Einstein Medical Center Philadelphia, 


BACKGROUND: Patients with end stage liver disease (ESLD) are 

a unique population prone to high hospital readmission rates, 

as evidenced by a 37% thirty day readmission rate (Volk M, 

2012) compared to a 20% readmission rate in the Medicare 

population of patients excluding those with liver disease. AIM: 

We tested an intervention designed specifically for patients with 

ESLD, intended to reduce 30 day readmission rates. METHODS: 

Using tools described in the evidence based literature for non- 

ELSD as a starting point, a liver specific discharge process was 

developed comprising 1) patient and family centered nurse 

administered medication training for adherence and side effect 

monitoring accompanied by performance feedback to assure 

understanding, 2) salt and free water restriction counseling, 

and 3) 48 hour post-discharge scripted telephone interviews 

for symptom and medication side effect monitoring, and follow 

up adherence. This process was implemented in June 2014. 

Nurses were educated to perform these interventions. Positive 

findings (e.g.; new symptoms) reported during the phone interview 

were managed with pre-determined responses, including 

electronic alerts to a physician for immediate intervention. 

Mean paired differences were compared for 30 day readmission 

rates during the six month period prior to implementation 

of the process, following a standard discharge process, and for 

the six months after implementation of the new discharge process. 

As this was a quality improvement exercise, all patients 

admitted to the Inpatient Liver Unit of the Einstein Medical Center, 

Philadelphia, were included. Results: The mean (SD) 30 

day hospital readmission rate for the six month period prior 

to implementation was 30.18(6.76), compared with 21.83 

(8.3) following implementation (p=0.19). Conclusion: In the 

ESLD population, strategies to reduce hospital readmissions


are needed. Our process demonstrated the possibility that a 

reduction in readmission rates could be achieved with a multifaceted 

process that incorporates nurse directed education with 

performance feedback and telephonic symptom assessment, 

coupled with rapid provider communications. A larger study 

will be required to confirm these findings. 

Disclosures: 

Simona Rossi - Consulting: BMS; Speaking and Teaching: gilead 

Eyob L. Feyssa - Advisory Committees or Review Panels: Gilead; Grant/Research 

Support: J&J, BMS, Abbot, Salix, Conatus, Cumberland, Merck; Speaking and 

Teaching: Gilead, BMS, Abbvie 

The following authors have nothing to disclose: Manisha Verma, Catherine Reynolds, 

Dee Morrison, June Smith, Cindy Mcglone, Victor J. Navarro 

531 

Thirty-day Readmission Following Liver Transplant in 

Medicare Beneficiaries – Incidence, Etiology, Outcomes, 

and Cost 

Lorna M. Dove 1 , Mary Jo Braid-Forbes 2 , Kevin F. Forbes 2 , Daniel 

J. Lerner 3 ; 1 Internal Medicine, Columbia University College of 

Physicians and Surgeons, New York, NY; 2 Braide-Forbes Health 

Research, Silver Spring, MD; 3 Health Sciences West, Scarsdale, 

NY 

BACKGROUND: Readmission of Medicare beneficiaries (MBs) 

within 30 days of hospitalization (30dRA) is considered a 

quality indicator, and it increases the total amount of annual 

Medicare spending. In 2015 the Centers for Medicare and 

Medicaid Services (CMS) can reduce reimbursements by up to 

3% for hospitals with high 30dRA rates for specific conditions. 

More than one-third of liver transplant (LT) recipients in the US 

are MBs, but little is known about 30dRA following LT in this 

group. PURPOSE: The purpose of this study is to define the 

incidence, etiology, outcomes, and cost of 30dRA following 

LT in adult MBs. METHODS: A total of 1,359 adult Medicare 

fee-for-service beneficiaries who underwent LT and survived 

to discharge in 2011 were identified and used to define the 

incidence of 30dRA following LT, and the principal diagnoses, 

outcomes, patient characteristics, and inpatient costs associated 

with those 30dRA. RESULTS: The incidence of 30dRA 

following LT in MBs was 37%. This is substantially higher than 

published 30dRA rates for MBs following acute myocardial 

infarction (AMI), heart failure (HF), pneumonia, and coronary 

artery bypass grafting (CABG). The most common principal 

diagnoses for 30dRAs were complications of a device implant 

or graft (21.5%) or medical or surgical care (12.9%), fluid and 

electrolyte disorders (5.4%), septicemia (4.4%), and acute/ 

unspecified renal failure (4.0%). The most common procedures 

at readmission were transfusion of blood products (11.7%), 

non-cardiac vascular catheterization (8.4%), and non-operating 

room GI procedures (8.2%). Factors associated with 

increased risk of readmission after risk adjustment were being 

Medicaid eligible (dual eligible) (OR 1.64, 95% CI [1.26- 

2.13]), and having renal failure at the time of transplantation 

(OR 1.44 95%CI [1.06-1.94]). The association of dual-eligibility 

with 30dRA is important because more than one-third 

of the MBs who underwent LT were dual-eligible, significantly 

more than in the general Medicare population. The mean cost 

of a 30dRA was $21,054. One year after hospitalization 

for LT, patients who had a 30dRA had significantly reduced 

survival (11.7% vs. 3.7%; P=0.01), and their total inpatient 

care costs were 34% higher, compared to patients without a 

30dRA ($252,985 vs. $189,352). CONCLUSIONS: In MBs, 

the 30dRA rate following LT was 37%, higher than reported 

for AMI, HF, pneumonia, or CABG. Complications were the 

most common cause for 30dRA. Patients with a 30dRA had 

decreased survival and increased inpatient costs at one year. 

Future research should focus on strategies to reduce complications 

from LT, and reduce 30dRA in patients at increased risk. 

Disclosures: 

The following authors have nothing to disclose: Lorna M. Dove, Mary Jo Braid- 

Forbes, Kevin F. Forbes, Daniel J. Lerner 

532 

Epidemiology and survival in patients with cirrhosis in a 

large commercial insurer database 

Steven J. Scaglione 1,3 , Leanne N. Metcalfe 2 , Stephanie Kliethermes 

3 , Rebecca Tsang 1 , Vik Goyal 4 , Allyce Caines 4 , Shaham 

Mumtaz 4 , Amy Luke 3 , Scott Cotler 1 ; 1 Hepatology, Loyola University 

Medical Center, Maywood, IL; 2 Enterprise Clinical Analytics, 

Health Care Services Corporation, Chicago, IL; 3 Preventive Medicine, 

Loyola University Medical Center, Maywood, IL; 4 Internal 

Medicine, Loyola University Medical Center, Maywood, IL 

Background: There are limited data regarding the epidemiology 

of cirrhosis in the general population (Scaglione, Volk, 

2014). We analyzed a large commercial insurance provider 

database to investigate the prevalence of cirrhosis, differences 

in survival in compensated versus decompensated cirrhosis, 

and differences in survival between NASH and HCV-related 

cirrhosis with and without diabetes. Methods: Retrospective 

matched case-control analysis estimating the prevalence in 

clinical-demographic, risk factors and survival associated with 

cirrhosis from Sept 2008-Aug 2013 by Health Care Services 

Corporation (HCSC), a commercial insurer providing healthcare 

coverage in 5 states. Cirrhosis was defined as ICD-9 code 

571.2, 571.5, or 571.6. The index date was the date of the 

first cirrhosis ICD-9-code occurring in the database after the 

first 6 months of continuous enrollment. Decompensated cirrhosis 

was defined as ICD-9 571.2, 571.5, or 571.6 AND a 

2nd diagnostic code for decompensation (any ICD-9: 070.44, 

070.71, 348.3x, 456.0, 456.1, 456.2x, 572.2, 572.3, 

572.4, 782.4, 789.59). Compensated cirrhosis is defined 

as those with ICD-9 571.2, 571.5, 571.6 and lacking ICD-9 

for decompensation as above. HCV Cirrhosis was defined by 

diagnostic codes for chronic HCV (ICD-9: 070.54 or 070.70) 

on at least two occasions a month or more apart. NAFLD 

was defined as by ICD-9 571.8 at any time in the absence 

of codes for other causes of liver disease. Results: During the 

study period, there were 18,202 persons with cirrhosis among 

3,478,093 enrollees, yielding a prevalence of 0.52%. Cirrhotics 

were older (55.6 vs 43.7,p


533 

Charges for Patients Hospitalized with Alcoholic Cirrhosis 

Exceed All Other Etiologies of Cirrhosis Combined 

and are Driven by Readmissions and Volume 

Monica Schmidt 1 , Paul H. Hayashi 2 , Ramon Bataller 2 , Alfred S. 

Barritt 2 ; 1 UNC Liver Center & Gillings School of Global Public 

Health, University of North Carolina Chapel Hill, Chapel Hill, NC; 

2 UNC Liver Center, Chapel Hill, NC 

Background:Total charges for hospitalized patients with cirrhosis 

are greater for alcoholic liver disease than for all other 

etiologies of cirrhosis combined. It is not known if costs are 

driven by a higher volume of hospitalizations, longer stays, 

or readmissions. Methods:We used the HCUP State Inpatient 

Database (SID) files for New York and Florida to determine 

costs, readmission rates, and predictors of 30-day readmission 

for patients with cirrhosis from 2010-2012. The SID 

provides longitudinal data for individual patients. A random 

effects model was used to evaluate predictors of 30 day readmissions. 

Results:215,886 discharges and 96,295 patients 

with cirrhosis were identified. Alcoholic cirrhosis accounted 

for 53% of total admissions and 51% of cirrhosis related 

inpatient charges, totaling $6B between 2010-2012. Costs 

per admission were $53,838 for alcohol and $56,326 for 

non-alcoholics. Alcoholics had greater aggregate charges at 

the index admission and 30, 60 and 90 days(Figure 1). 30 

day readmission rates were 14% for alcoholics and 12% for 

non-alcoholics(p 

31 IU/L in women, ALT> 40 IU/L or AST> 37 IU/L in men) 

and excessive alcohol use (>20 g/day in men, >10 g/day in 

women). Hepatitis C (HCV) infection was defined as detectable 

HCV RNA. Presumed NAFLD was defined as elevated liver 

enzymes in the absence of indicators of chronic hepatitis B 

and C infection (negative antibody serology) and no excessive 

alcohol use. Type 2 diabetes (DM) was defined as the use of 

glucose-lowering agents or fasting blood glucose >125 mg/dL. 

The NHANES medical history questionnaire was used to determine 

the presence of major chronic diseases (cardiovascular, 

pulmonary, renal, malignancies) which were further tested as 

predictors of mortality in a Cox proportional hazard model. 

Results: In 1999-2012, 11,726 BB subjects [age at enrollment 

50.2±0.3 years, 72.5% White, 10.9% Hispanics, 11.4% African-American, 

36.5% obese (BMI≥30), 10.6% with DM] were 

included. The most common cause of CLD in BB was NAFLD 

(11.08% of BB population), followed by ALD (2.82%) and HCV 

(2.27%). These prevalence rates in the leading-edge BB and 

late BB were: NAFLD: 10.4±0.6% vs. 11.7±0.6%, p=0.11; 

ALD: 2.0±0.3% vs. 3.5±0.4%, p=0.0044, HCV: 2.8±0.3% vs. 

3.6±0.4%, p=0.09. During average 92.4 months of follow-up, 

355 (3.84%) BB participants with mortality data died. Multivariate 

analysis showed that HCV infection [adjusted hazard ratio 

(aHR) = 2.06 (1.03-4.11)] was independently associated with 

increased mortality in BB. Additionally, older age [aHR = 1.05 

(1.02-1.08) per year], DM [HR = 1.90 (1.21-2.98)], hypertension 

[aHR = 2.19 (1.58-3.04)], excessive alcohol use [aHR 

= 2.62 (1.73-3.97)], the presence of cardiovascular diseases 

[aHR = 1.70 (1.06-2.72)], and chronic kidney disease [aHR = 

3.46 (1.87-6.39)] were associated with mortality in BB population. 

Conclusions: NAFLD, ALD and HCV are common causes 

of CLD in BB. However, only HCV remains an independent 

CLD predictor of mortality in BB cohort. As more HCV patients 

are being cured, NAFLD will most likely account for most of the 

CLD burden in the BB cohort. 

Disclosures: 






Inc.


The following authors have nothing to disclose: Maria Stepanova, Sarah Elfeky, 

Alexandrea Srishord, Carey Escheik, Leo Mclaughlin, Mariam Afendy, Robert J. 

Wong, Sharon L. Hunt 

535 

A New Approach to Patient Centered Care in Hepatology: 

Patient Reported Outcomes Assessment 

Manisha Verma 1 , Victor J. Navarro 1 , Shana D. Stites 2 , Eyob L. 

Feyssa 1 , Simona Rossi 1 ; 1 Hepatology, Einstein Healthcare Network, 

Philadelphia, PA; 2 Center for Urban Health, Einstein Healthcare 

Network, Philadelphia, PA 

Background: Patient Reported Outcomes (PRO) are measures of 

symptoms, health behavior &/or experiences reported directly 

by patients. Integrating PRO Assessment (PA) in routine care 

improves patient centered care (PCC). PROMIS-29 (Patient 

Reported Outcomes Measurement Information System) is a 

web-based NIH supported validated set of 29 questions used 

for PA. Our aim is to identify if PA influences PCC in patients 

with cirrhosis, through enhanced communication, & facilitating 

detection of physical/ psychological issues. Methods: This is 

a prospective nonrandomized study of the feasibility, acceptability, 

& clinical value of PA in cirrhotics. PA included 1) 

provider training in PA score interpretation, 2) administering 

PROMIS-29, & 3) reporting PA results (T-scores) to patients & 

providers for point of care use. T-score measures the symptom 

assessed. Higher scores reflect greater symptom assessed. All 

outpatients with cirrhosis were eligible. Patient and provider 

feedback surveys were administered after each visit to assess 

acceptability/feasibility. Descriptive statistics were used to 

assess baseline measures; paired t-tests were used to compare 

baseline and 3 month data. Results: 50 patients were enrolled; 

3 month data were available for 20. There were no statistically 

significant differences in baseline measures based on 

gender, race, or duration of liver disease (p>0.05). At baseline, 

pain, anxiety, & fatigue were worse in our sample vs the 

US population (p


537 

Physician versus Patient Perceptions of Medical Care 

Quality in Primary Biliary Cirrhosis 

Andrew Saich, Herbert Swanson, Tracy J. Mayne; Intercept Pharmaceuticals, 

Inc., San Diego, CA 

Numerous studies have demonstrated significant self-enhancement 

bias in physician perceptions of medical care delivery, in 

which physicians rate care delivery more highly than patients. 

Objective: To compare patient and physician perceptions of 

care in patients with primary biliary cirrhosis (PBC) and the 

hepatologists and gastroenterologists treating PBC patients, 

with the goal of improving care delivery. Methods: From 

December 11, 2014 to January 12, 2015, we conducted surveys 

with board certified gastroenterologists (262) and hepatologists 

(60) practicing for at least two years and had treated 

at least two PBC patients in the past six months. From January 

5-30, 2015, we conducted surveys with 214 patients with PBC 

who were under a physician’s care. Results: The figure shows 

physician and patient ratings of their confidence in physician 

delivery of 7 areas of PBC care. Only 2 areas differed by more 

than 5 points: physicians under-rated their skills in helping 

patients manage PBC symptoms by a mean 6 points, and overrated 

their bedside manner by a mean 10 points, compared to 

patient ratings. Beyond care provisions, there were meaningful 

differences between patients and physicians perceptions on 

how PBC affects patient health-related quality of life. Half of 

PBC patients reported that PBC impacts a “great deal” how 

they feel physically (50% vs. 25% physicians), their ability 

to work (39% vs. 16% physicians), and their ability to do the 

things they enjoy (36% vs. 17% physicians). Despite the impact 

of PBC on these symptoms, only 42% of patients strongly agree 

that additional treatment options would treat PBC more effectively 

(versus 65% of physicians). Conclusion: Hepatologists 

and gastroenterologists do not demonstrate systematic self-enhancement 

bias in treating PBC patients. While they may overrate 

their bedside manner, they underrate their management 

of PBC symptoms. However, physicians and patients both 

rated diagnosis and treatment more highly than monitoring 

disease progression and managing symptoms, and physicians 

recognize the need to improve practice. If physicians’ estimation 

and management of patient quality of life improves, then 

patient perception of the treatment effectiveness and optimism 

toward treatment innovation may also improve. Interventions to 

improve physician communication around disease progression, 

and patient communication around symptoms and symptom 

management could enhance practice. 

Disclosures: 



The following authors have nothing to disclose: Andrew Saich 

538 

Redesigning Outpatient Care to Reduce 30-day Hospital 

Readmissions Among Recent Liver Transplant Recipients. 

Chanda Ho, Nina Topic, Raphael Merriman, Robert W. Osorio, 

Garrett M. Hisatake, R Todd Frederick; California Pacific Medical 

Center, San Francisco, CA 

Background: Hospital readmission after liver transplantation 

(LT) is associated with reduced patient and graft survival and 

represents a growing health care burden. Prevention of hospital 

readmission has been an area of focus by the Centers 

for Medicare and Medicaid Services with reduced payments 

for readmissions for certain diagnoses. However, hospital 

readmissions and strategies to reduce them among recent LT 

recipients have not been well characterized. Methods: We 

collected baseline hospital readmission data following LT to 

our medical center from 2010-2012. Reasons for readmission 

were reviewed by a hepatologist and a surgeon. We captured 

median length of stay (LOS) both after LT and during readmission. 

Based on these data, in October 2012, we implemented 

a quality improvement project to reduce our readmission rates. 

Readmission data post-intervention were collected for 2013 

and 2014. Results: Our 30-day post-LT hospital readmission 

rates in 2010, 2011, and 2012 were 40%, 42%, and 43% 

respectively. A high proportion of readmissions had a short 

LOS (≤ 2days):47% and 48% in 2011 and 2012, respectively. 

The reviewers determined a significant proportion of these 

admissions could have been managed in the outpatient setting. 

Barriers to care management identified included difficulty 

scheduling multiple, often complex procedures in the outpatient 

setting. The organization of post transplant care involving multiple 

expedited procedures (e.g. interventional radiology, endoscopy) 

was streamlined to optimize outpatient care coordination 

and delivery for the commonly required diagnostic and therapeutic 

procedures (e.g. ERCP, liver biopsy, ultrasound). Thirty-day 

readmission rates declined in 2013 and 2014 to 25% 

and 26%, respectively (p


and 1965. However, many eligible patients are not being 

screened. Aim: Determine the rate adherence to the CDC 

recommendations, and evaluate differences in demographics 

between those screened to those who were not. Methods: 

Single center-center, retrospective cohort study of all patients 

born between 1945-1965 who were seen at the Primary Care 

Group (PCG), Emergency department (ED), Infectious Disease 

(ID), Gastroenterology Department (GI) from August 17, 2012 

to November 4, 2014 were included in the study. Insurance, 

race, results of HCV related laboratory testing were included 

in the study. Results: A total of 24965 patients (10 499 males) 

born between 1945 and 1965 and no known history or documented 

HCV screening history were seen at the University of 

Chicago’s ED, PCG, ID and GI. Of the 24 965 eligible individuals 

2,104 were screened for chronic HCV (889 males). 

Unscreened individuals were younger compared to screened 

(56 vs. 58, respectively (p


reflect the position or policy of the Department of Veterans 

Affairs or the United States government. 

Disclosures: 

Christine M. Hunt - Consulting: Otsuka 

Ayako Suzuki - Consulting: Novartis 

Hans L. Tillmann - Consulting: Novartis; Employment: AbbVie; Grant/Research 

Support: Novartis; Stock Shareholder: AbbVie, Abbott, Gilead 

Joseph K. Lim - Consulting: Merck, Boehringer-Ingelheim, Gilead, Bristol Myers 

Squibb, Janssen; Grant/Research Support: AbbVie, Boehringer-Ingelheim, Bristol 

Myers Squibb, Gilead, Hologic, Janssen 

The following authors have nothing to disclose: Lauren A. Beste, George N. 

Ioannou, Elliott Lowy, Dawn T. Provenzale, Michael J. Kelley, Cynthia A. Moylan, 

Omobonike O. Oloruntoba, David Ross 

541 

Comparable hepatitis C treatment outcomes with oral 

direct-acting antivirals between rural outreach clinics 

and an academic medical center: A case for expanding 

task-shifting 

Channa R. Jayasekera, Ryan B. Perumpail, Aijaz Ahmed; Division 

of Gastroenterology and Hepatology, Stanford University Medical 

Center, Stanford, CA 

Purpose: Residence in medically underserved areas is associated 

with lower likelihood of receiving hepatitis C treatment. 

The safety, tolerability, and effectiveness of oral direct-acting 

antivirals (DAA) may obviate the need for specialist expertise 

in many patients, and decentralizing treatment from specialists 

to more abundant community-based providers may improve 

treatment access without compromising safety. At our rural 

outreach clinics (OCs), a visiting hepatologist evaluates treatment 

eligibility but subsequent routine management is devolved 

to licensed vocational nurses (task-shifting). The hepatologist 

reviews laboratory results remotely and is available for consultation 

and as-needed clinic visits. We assessed the outcomes 

of patients treated at these OCs versus those treated directly 

by the hepatologist at an academic medical center (AMC). 

Methods: We retrospectively compared clinical characteristics, 

sustained virological response 12 weeks after treatment completion 

(SVR12) rates, treatment discontinuation and adverse 

event rates of consecutive patients treated with DAAs by a 

single hepatologist between Dec 2013-Sept 2014 at an AMC 

and three OCs. Results: 45 AMC and 48 OC patients received 

either sofosbuvir+ribavirin (SOF+RBV) or sofosbuvir+simeprevir 

(SOF+SMV). AMC and OC patients were comparable in 

age, gender, HCV genotype distribution, and prior treatment 

experience. More AMC patients were cirrhotic (73.3% vs 

50%, p=0.03) and received SOF+SMV (77.8% vs. 56.3%, 

p=0.03). Overall SVR12 rates were comparable between the 

AMC and OCs (88.8% vs. 83.3%, p=0.87), as were SVR12 

rates of non-cirrhotic, cirrhotic, treatment-naïve, treatment-experienced, 

genotype 1, 2, and 3 patients (figure 1). One patient 

discontinued treatment at each site. There were no treatment 

related hospitalizations or deaths. Conclusions: In this analysis 

of oral DAA therapy at rural outreach clinics and an academic 

medical center, treatment responses were comparable to published 

real-world data, and site of treatment or type of provider 

did not impact treatment outcome. This task-shifting strategy, 

wherein mid-level providers are empowered to manage HCV 

treatment in medically underserved areas with remote supervision 

of specialists, may hold promise in expanding HCV treatment 

access. 

Disclosures: 




Inc. 

The following authors have nothing to disclose: Channa R. Jayasekera, Ryan B. 

Perumpail 

542 

Reducing Wait Times for Radiofrequency Ablation in 

Patients with Hepatocellular Carcinoma: A Quality 

Improvement Initiative at the Toronto Centre for Liver 

Disease. 

Mayur Brahmania 1 , Osman Ahmed 1 , Korosh Khalili 2 , Eberhard L. 

Renner 1 , Harry L. Janssen 1 , Morris Sherman 1 , David K. Wong 1 , 

Hemant Shah 1 , Jordan J. Feld 1 ; 1 Gastroenterology, University of 

Toronto, Toronto, ON, Canada; 2 Medical Imaging, University of 

Toronto, Toronto, ON, Canada 

Introduction: Radiofrequency ablation (RFA) has become the 

recommended curative treatment option for patients with early 

stage hepatocellular carcinoma (HCC). Unfortunately, the 

expected number of HCC cases requiring RFA will grow at a 

greater rate than potential RFA capacity. Thus, novel strategies 

will be required to handle the burden. According to guidelines, 

waiting times for any malignancy from diagnosis to treatment 

should not exceed 30 days. In the current analysis we investigated 

if wait times for RFA were associated with outcomes such 

as residual disease after RFA, tumor recurrence, liver transplant 

or death. Methods: This was a retrospective study analysing 

patients with HCC between January 2012 and December 

2014 presenting to University Health Network (UHN) hospitals 

in Toronto. Multi-disciplinary Tumor Board rounds were held 

weekly on patients with newly diagnosed HCC. All patients 

receiving RFA alone as treatment for HCC were included. The 

time from diagnosis to presentation at Tumor Board rounds 

and the time from Tumor Board rounds to the date the patient 

received first treatment was documented. Multivariable Cox 

regression was used to determine factors associated with tumor 

outcome. Results: Of the 150 patients included in the study, 

82% percent were male and the median age was 62 years 

(Range 25-90). Median tumor size at diagnosis was 19 mm 

(Range 0-43 mm), mean MELD was 7 (SD=4.3) and 58% had 

Barcelona stage A. The cause of liver disease was hepatitis C 

in 41%, hepatitis B in 30%, Hepatitis B/C co-infection in 2%, 

and other liver diseases in 27%. The median wait time from 

diagnosis to Tumor Board presentation was 21 days (range 

12-41), wait time from Tumor Board to Interventional Radiology 

consultation was 30 days (21-48) and the mean time from 

consultation to RFA was 44 days (IQR 34-58). The median 

time from HCC diagnosis to RFA treatment was 98 days (IQR 

79-139). 30 patients died or underwent transplant. When 

adjusting for Barcelona staging, tumor size and MELD score, 

wait times >150 days was not associated with an increased 

risk of death or transplant (HR=1.69; p=0.23). Conclusion: 

Our study demonstrates wait times for RFA in patients with HCC 

are over the recommended waiting time in our province. Wait 

times over 150 days showed a non-significant trend towards


an increased risk of transplant or death. We have identified 

system gaps where quality improvement measures can be 

implemented to reduce wait times and allocate resources for 

future RFA treatment at our centre. 

Disclosures: 

Eberhard L. Renner - Advisory Committees or Review Panels: Vertex Canada, 

Novartis, Astellas Canada, Roche Canada, Gambro, AbbVIe, BMS, Gilead; 

Grant/Research Support: Novartis Canada, Gilead; Speaking and Teaching: 

Novartis, Astellas Canada, Roche Canada 






Morris Sherman - Consulting: Gilead, Merck, Bayer, Arqule, Celsion, Boehringer 

Ingelheim, Janssen, Abbvie; Grant/Research Support: WAKO; Speaking and 

Teaching: WAKO 

David K. Wong - Grant/Research Support: Gilead, BMS, Vertex, BI 

Hemant Shah - Consulting: Merck, Roche, Gilead, Janssen, BMS, Abbvie, Lupin 




The following authors have nothing to disclose: Mayur Brahmania, Osman 

Ahmed, Korosh Khalili 

543 

Timing of Hepatitis B and C Diagnosis Relative to Hepatocellular 

Carcinoma Diagnosis in the BC Hepatitis Testers 

Cohort (BC-HTC) 

Naveed Z. Janjua 1,2 , Amanda Yu 1 , Margot E. Kuo 1 , Maryam 

Alavi 4 , Ryan Woods 3 , Maria Alvarez 1 , Gregory Dore 4 , Mark Tyndall 

1,2 , Mel Krajden 1,2 ; 1 BC Centre for Disease Control, Vancouver, 

BC, Canada; 2 University of British Columbia, Vancouver, BC, 

Canada; 3 BC Cancer Agency, Vancouver, BC, Canada; 4 University 

of New South Wales, Sydney, NSW, Australia 

Background: Early diagnosis of hepatitis B (HBV) and hepatitis 

C (HCV) followed by treatment could prevent late stage liver 

disease. Hepatitis diagnosis after appearance of chronic liver 

disease sequelae is an indicator of sub-optimal screening. We 

measured the timing of HBV and HCV diagnoses relative to 

detection of hepatocellular carcinoma (HCC) as an indicator of 

late hepatitis diagnosis. Methods: The British Columbia Hepatitis 

Testers Cohort (BC-HTC) consists of 1,417,780 individuals 

tested for HCV, HIV and/or reported to public health as a case 

of HBV, HCV, HIV or tuberculosis from 1990-2013. Cohort 

data were linked with medical visits, hospitalizations, cancers, 

prescription drugs and mortality. Late hepatitis diagnosis was 

defined as HBV or HCV diagnosis within 2 years prior to or 

after HCC diagnosis. HCC diagnosis was based on ICD-O-3 

topography code C22.0 and histology codes 81703-81753 in 

the BC Cancer Registry. HCV diagnosis date was the date of 

the first positive test. HBV diagnosis date was the date HBV was 

reported to public health. Hepatitis diagnoses from 1990-2012 

and HCC from 1992-2011 were analyzed to allow for a 2 

year lead-in and one year follow-up time between hepatitis and 

HCC diagnoses. Results: Between 1992 and 2011, 2,250 

cases of HCC were diagnosed; 612 (27%) were HBV positive, 

868 (39%) HCV positive, and 47 (2%) had both. Overall, 

659/34,775 HBV cases (1.9%) and 915/65,842 HCV cases 

(1.4%) developed HCC. Among HBV and HCV cases with 

HCC, 45% and 31% respectively were diagnosed late. HBV 

late diagnosis declined from 100% in 1992 to 53% in 2000 

to 24% in 2011. HCV late diagnosis declined from 100% in 

1992 to 30% in 2000 to 13% in 2011. In the multivariable 

logistic regression model, birth years


both tests, the correct answer was chosen 79% of the time after 

the rotation. Conclusion: Our novel curriculum for a non-elective 

consult rotation has effectively demonstrated improvement in IM 

residents’ comfort with and knowledge of CLD. A trend towards 

increased career interest in hepatology was also observed, 

which suggests more exposure to CLD could positively impact 

recruitment to the workforce. 

Disclosures: 

Donald M. Jensen - Advisory Committees or Review Panels: Merck, Onom Foundation; 

Speaking and Teaching: Gilead 










The following authors have nothing to disclose: Adam E. Mikolajczyk, Jeanne M. 

Farnan, John F. McConville, Andrew I. Aronsohn 

545 

High Rates of Hepatitis C Virus Infection Among Inpatient 

Baby Boomers in an Urban Hospital: a model to 

improve linkage to care 

Aparna Goel 1 , Lismeiry Paulino 2 , Douglas Dieterich 2 , Ponni V. 

Perumalswami 2 ; 1 Gastroenterology, Icahn School of Medicine 

Mount Sinai, New York, NY; 2 Liver Diseases, Icahn School of 

Medicine Mount Sinai, New York, NY 

Background: New York State passed legislation that took effect 

in January 2014 requiring healthcare providers to offer Hepatitis 

C Virus (HCV) screening to all inpatient baby boomers 

and link infected persons to care. Our aims are to determine 

the prevalence of HCV-positive baby boomers admitted to an 

urban, tertiary care hospital and evaluate the sequential impact 

of an electronic medical record (EMR) prompt, provider education, 

and data feedback to providers on HCV screening 

and linkage to care rates. Methods: The results of HCV antibody 

(Ab) tests for individuals born 1945-1965 with inpatient 

encounters at Mount Sinai were obtained from November 

2013 through April 2015. The type of medical service, level 

of provider and patient demographics were collected. HCV 

screening and linkage to care rates before and after a series 

of interventions were compared: a birth cohort-selective EMR 

prompt was implemented in January 2014, comprehensive 

education for housestaff and nurses occurred from July-September 

2014, monthly data feedback to nurses with screening 

rates began in October 2014 and finally patient navigation 

was implemented in May 2015. Results: There were 23,835 

inpatient baby boomer encounters during the study period. 

The median age was 60 years with 56% male and 33% Hispanic. 

Overall, 34% had a documented HCV Ab test and 

13% (n=1,036) were positive. Screening rates did not significantly 

rise following an EMR prompt (p=0.92), education 

(p=0.86) or with data feedback to nursing managers (p=0.42). 

Provider education and data feedback interventions led to a 

rise in newly diagnosed HCV-positive rates from 6% to 15% 

(p576,000 

patients receiving AMA testing between January 2010 and 

December 2013 in a large US commercial laboratory database. 

Patients were considered AMA positive if the antibody 

titer was > 1:20 or M2 antibody was > 25.0 U. We examined 

geographic distribution at a 3-zip code level. Results: 16,492 

patients tested AMA positive and had at least 1 ALP measure 

over the 4 year period. 5,380 had an ALP >120 U/l before or 

within 30 days of their first AMA+ test and were classified as 

having probable PBC. There were clear patterns of increased 

concentration of incident PBC patients (from west to east): 

Southeastern tip of the Alaskan spur; Santa Barbara/Ventura 

and inland central California; western Dakotas’ border; central 

Oklahoma including Oklahoma City; western Texas and Houston; 

Indiana just north of Louisville, Kentucky; an area of the 

Appalachians spanning southeastern New York, southern Pennsylvania, 

and the northern West Virginia border; and Florida 

(excluding the Pan Handle). These areas were not associated 

with age, race/ethnicity, urban concentration, or elevation. 

Conclusion: These analyses indicate that there are clusters of 

incident patients in geographically diverse areas in the United 

States. Additional analyses are required to determine if there 

may be geographic or demographic factors underlying these 

variations.


Disclosures: 







The following authors have nothing to disclose: Keith D. Lindor 

547 

A qualitative assessment of factors impacting adoption 

and implementation of USPSTF age-based Hepatitis C 

Virus screening guidelines 

Amy B. Jessop; HepTREC, University of the Sciences in Philadelphia, 


Background: Baby boomers comprise 70% of U.S. Hepatitis 

C (HCV) cases, yet many remain unaware of their infection. 

The 2012 CDC and 2013 USPSTF HCV screening guidelines 

that expanded to include one-time screening for those born 

between 1945 and 1965 were developed to increase identification 

of those infected. Insurer policies require or encourage 

screening within primary care or family practice settings, 

therefore, improvements will be realized only if primary care 

providers adopt the guidelines. This qualitative study examines 

the awareness and adoption of prevention practice guidelines, 

particularly the enhanced USPSTF HCV screening guideline in 

a large, suburban family practice setting. Methods: Data collection 

employed individual interviews conducted at the practice 

site using a semi-structured interview guide developed, 

assessed for content validity through literature and expert 

review and pilot testing with three practicing primary care 

physicians. The family practice site selected for examination 

is a community-based family practice and resident training 

program. Interviews with all resident (12) and attending physicians 

(5) and advanced practice nurse (NP) were taped and 

completed between Dec 2014 and Feb 2015. They considered 

knowledge and beliefs about practice guidelines (in general 

and the USPSTF enhanced HCV guideline in particular) and 

self-assessed screening rates. Researchers independently, then 

collectively, reviewed transcriptions, assessing responses and 

identifying common and emergent themes. Results: All subjects 

viewed practice guidelines as useful tools. Attending physicians 

reported self-selection of guidelines to adopt and implement 

while residents tended to follow advice of the attending 

physicians to which assigned. Source of the guideline played 

a deciding role in which guidelines were adopted. Awareness 

of the expanded USPSTF HCV screening guideline varied by 

position, with the NP and attending physicians most aware, 

although knowledge of guideline details was limited; some 

considered their patients to be of low-risk for HCV infection 

and estimates of individual and group HCV screening rates 

were low (2-25%). Subjects caring for an HCV-affected patient 

reported greater importance to screening. Smart tools, prompting 

screening at point of care were deemed likely to improve 

practice. Conclusion: Preventive practice guidelines are well 

accepted, but knowledge of and attitudes about age-based 

HCV screening guidelines is low and lack of reminders at pointof-care 

limit screening initiation. Improved education and systems-based 

approaches to improve screening practices are 

essential to reap benefits of expanded guidelines. 

Disclosures: 

The following authors have nothing to disclose: Amy B. Jessop 

548 

Outreach invitations improve HCC surveillance rates: 

A Randomized Controlled Trial in a Safety-Net Health 

System 

Amit G. Singal 1 , Jasmin A. Tiro 2 , Katharine McCallister 2 , Caroline 

A. Mejias 2 , Lei Xuan 2 , Ethan Halm 1 ; 1 Internal Medicine, University 

of Texas Southwestern, Irving, TX; 2 University of Texas Southwestern, 

Dallas, TX 

Background: Hepatocellular carcinoma (HCC) surveillance 

is associated with improved survival, but its effectiveness is 

limited by underuse, particularly in underserved populations. 

Aim: Compare the effectiveness of mailed outreach and patient 

navigation to increase HCC surveillance rates in a racially 

diverse cohort of patients Methods: Patients with documented 

or suspected cirrhosis at a safety-net health system were randomized 

to receive mailed outreach invitations for surveillance 

ultrasonography, mailed outreach plus patient navigation, or 

usual care. Documented cirrhosis was defined using ICD-9 

codes, and suspected cirrhosis was defined as AST to platelet 

ratio index (APRI) ≥1.5 in the presence of liver disease. We 

excluded those with Child C cirrhosis who were not transplant 

candidates and/or significant comorbid conditions given limited 

benefit of surveillance. Patients who did not respond to 

outreach invitations within 2 weeks received up to 3 reminder 

telephone calls. The primary study outcome was completion of 

surveillance ultrasound within 6 months of randomization. We 

used intent- to-screen principle for analyses. We are randomizing 

1800 patients in groups of 300 (100 patients/arm), with 

results of the first 2 groups presented here. Results: Baseline 

patient characteristics (n=600) across the arms were similar. 

Mean age was 55 years and 60% were men. The cohort was 

racially diverse with 36% Black, 35% Hispanic, and 28% 

White. Most (79%) had documented cirrhosis, while 21% had 

suspected cirrhosis. Cirrhosis was due to HCV in 54%, alcohol 

19%, NASH 15%, and HBV 4%. Rates of scheduled surveillance 

ultrasound were significantly higher in the outreach/navigation 

(34.0%) and outreach alone (26.5%) arms than usual 

care (11.5%; p


549 

Engagement in care of high risk hepatitis C patients 

with interferon-free therapies 

John Dever 1,2 , Julie Ducom 1 , Ariel Ma 1 , Michael Yang 1 , Erika 

Cherk 1 , Ann Herrin 1 , Erik J. Groessl 1,2 , Samuel B. Ho 1,2 ; 1 VA San 

Diego Healthcare System, San Diego, CA; 2 University of California, 


Objectives: New interferon-free direct acting antiviral (DAA) 

treatments have the potential to expand the numbers of patients 

receiving treatment for hepatitis C virus (HCV); however patient 

access and willingness to engage in care is unknown. Our 

aim was to determine patient engagement and patient-related 

barriers to care for accessing new interferon-free DAA in a realworld 

practice setting. Methods: Patients with viremic HCV and 

high-risk for fibrosis by FIB-4 score were identified using the 

national VA HCV registry within the San Diego VA Healthcare 

System in October 2014. Patients were categorized as actively 

involved in HCV clinic care or those who were not seen in 

HCV clinic within the previous 6 months and without a future 

appointment. Patients were contacted by letter and phone call 

and notified of the availability of new more effective DAA treatments 

and the consequences of not receiving treatment, and 

invited to schedule an appointment in HCV clinic. Patient characteristics 

and subsequent outcomes were determined using 

medical records. Logistic regression was used to examine factors 

associated with willingness to engage in care. Results: 

481 patients were found to have FIB-4 > 2.4 (307 with FIB-4 

> 3.25 with high likelihood of cirrhosis). Of the 481 patients, 

177 patients (37%) were actively followed in clinic and eligible 

for treatment. Treatment ineligible included 24 (5%) in 

hospice or deceased, 66 (14%) relocated, and 12 (2%) with 

severe comorbidity. 202 treatment eligible patients (42%) were 

identified as never seen by a HCV clinic provider or lost to follow-up. 

Of these, 123 (61%) remained non-engaged and gave 

no response to outreach (follow up 2-6 mo), 6 patients (3%) 

declined evaluation and treatment, and 74 (36%) responded 

and engaged with HCV clinic. Non-engaged patients compared 

with engaged were more likely to have been homeless 

(48 vs 28%), have active alcohol/drug use (44 vs 31%), psychiatric 

diagnoses (69 vs 63%), and # co-morbid disorders 

(3.17 vs 3.07). Significant variables in logistic regression for 

non-engagement included history of homelessness, COPD, and 

hypertension. Conclusions: Among treatment-eligible, high priority 

HCV patients most at risk for advanced fibrosis that were 

not currently attending HCV clinic, 64% were unable or unwilling 

to engage in HCV care after a one time outreach (32% of 

the total treatment eligible). Homelessness and certain comorbidities 

are significant factors related to non-engagement. 

These data indicate that more sustained or intensive outreach 

efforts are needed in order to effectively expand access to 

many of those most in need of treatment. 

Disclosures: 

Samuel B. Ho - Grant/Research Support: Genentech, Gilead; Speaking and 


The following authors have nothing to disclose: John Dever, Julie Ducom, Ariel 

Ma, Michael Yang, Erika Cherk, Ann Herrin, Erik J. Groessl 

550 

Thirty-day Readmission for Infection Following Liver 

Transplant is Associated with Increased Mortality 

Lorna M. Dove 1 , Daniel J. Lerner 2 ; 1 Internal Medicine, Columbia 

University College of Physicians and Surgeons, New York, NY; 

2 Health Sciences West, Scarsdale, NY 

BACKGROUND: Over thirty percent of patients are readmitted 

within 30 days of liver transplantation (30dRA). The role of 

infection in 30dRA of these immunosuppressed patients is not 

well defined. PURPOSE: The purpose of this study is to define 

the incidence, etiology, and cost of 30dRA for infection following 

liver transplantation (LT). METHODS: Utilizing the Agency 

for Healthcare Research and Quality’s Healthcare Cost and 

Utilization Project’s State Independent Databases for 4 geographically 

distinct states: Florida, Massachusetts, New York, 

and Washington, we identified patients who underwent LT in 

2012, based on the ICD-9-CM code for LT, 50.59. Patients 

with a 30dRA following LT were divided into 2 groups: those 

with and without an infection-related principal diagnosis (IRPD) 

for the 30dRA. Patients with a 30dRA for rehabilitation on the 

day of discharge were excluded. Approximately 8% of 30dRA 

were not captured because they occurred in the following 

year. RESULTS: The incidence of 30dRA following LT was 30% 

(332/1099). Nineteen percent (64/332) of those 30dRA had 

an IRPD. There was no significant difference in the mean age, 

gender, race, number of chronic conditions, primary payor, or 

median household income between the two groups. There were 

21 different IRPD, and the most common were postoperative 

infection (31%; 20/64), intestinal infection due to C. difficile 

(16%; 10/64), unspecified septicemia (13%; 8/64), and urinary 

tract infection (8%; 5/64). Patients with a 30dRA for an 

IRPD were more likely to die during that 30dRA, compared 

to those with a non-IRPD (5% vs. 0%; P


551 

Evacuation after the Fukushima Daiichi Nuclear Power 

Plant accident as a cause of liver dysfunction: The 

Fukushima Health Management Survey 

Atsushi Takahashi 1,2 , Tetsuya Ohira 2,3 , Mitsuaki Hosoya 2,4 , Seiji 

Yasumura 2,5 , Masato Nagai 2,3 , Hiromasa Ohira 1 , Shigeatsu 

Hashimoto 2,6 , Hiroaki Satoh 2,6 , Akira Sakai 2,7 , Akira Ohtsuru 2,8 , 

Yukihiko Kawasaki 2,4 , Hitoshi Suzuki 2,9 , Gen Kobashi 10 , Kotaro 

Ozasa 11 , Shunichi Yamashita 2,12 , Kenji Kamiya 2,13 , Masafumi 

Abe 2 ; 1 Gastroenterology and Rheumatology, Fukushima Medical 

University School of Medicine, Fukushima, Japan; 2 Radiation Medical 

Science Center for the Fukushima Health Management Survey, 

Fukushima Medical University School of Medicine, Fukushima, 

Japan; 3 Epidemiology, Fukushima Medical University School of 

Medicine, Fukushima, Japan; 4 Pediatrics, Fukushima Medical 

University School of Medicine, Fukushima, Japan; 5 Public Health, 


Japan; 6 Nephrology, Hypertension, Diabetology, and Endocrinology, 

Fukushima Medical University School of Medicine, 

Fukushima, Fukushima, Japan; 7 Radiation Life Sciences, Fukushima 

Medical University School of Medicine, Fukushima, Japan; 8 Radiation 

Health Management, Fukushima Medical University School 

of Medicine, Fukushima, Japan; 9 Cardiology and Hematology, 


Japan; 10 Public Health, Dokkyo Medical University, Tochigi, 

Japan; 11 Epidemiology, Radiation Effects Research Foundation, 

Hiroshima, Japan; 12 Japan and Atomic Bomb Disease Institute, 

Nagasaki University, Nagasaki, Japan; 13 Research Institute for 

Radiation Biology and Medicine, Hiroshima University, Hiroshima, 

Japan 

Background and Aim: The Great East Japan Earthquake and 

Fukushima Daiichi Nuclear Power Plant accident caused residents 

to forego their normal lives and evacuate. The aim of 

this study was to evaluate liver function before and after this 

disaster. Methods: This was a longitudinal survey of 26,006 

Japanese men and women living near the Fukushima Daiichi 

Nuclear Power Plant in Fukushima prefecture. This study was 

undertaken using data from annual health checkups of persons 

40-90 years old from 2008 to 2010. Follow-up examinations 

were conducted from June 2011 to March 2013, with a 

mean follow-up of 1.6 years. Changes in liver function before 

and after the disaster were compared among evacuees and 

non-evacuees. Liver dysfunction was defined as aspartate aminotransferase 

≥40 IU/L, alanine aminotransferase ≥40 IU/L, or 

γ-glutamyl transpeptidase ≥50 IU/L. Subjects were divided into 

three alcohol intake groups: 1) non-drinkers, defined as drinking 

no alcohol; 2) light drinkers, defined as


preventive liver care measures for IBD patients 

Disclosures: 










The following authors have nothing to disclose: John N. Gaetano, Andrew I. 

Aronsohn 

Disclosures: 

Robert E. Smith - Speaking and Teaching: Gilead, Salix, Abbvie, Janssen 

The following authors have nothing to disclose: Chuan L. Miao, Sara West, Korta 

Yuasa, Michael Komar, Stacy Prall 

553 

Disparities in the Quality of Death and Resource Utilization 

in End-Stage Liver Disease 

John N. Gaetano, K. Gautham Reddy, Andrew I. Aronsohn, Helen 

S. Te, Nancy Reau; Department of Medicine, University of Chicago 

Hospitals, Chicago, IL 

Background: Various studies have shown increased utilization 

of life-sustaining measures as well as increased end-of-life costs 

in minorities. It is unknown if this disparity exists in the end-of life 

care among patients with end-stage liver disease. Methods: The 

Nationwide Inpatient Sample from 2012 was used to capture 

patients with cirrhosis and a manifestation of hepatic decompensation 

(variceal bleeding, hepatorenal syndrome, ascites, 

spontaneous bacterial peritonitis, or portosystemic encephalopathy), 

above the age of 39, who died in the hospital. The 

primary predictor variable was race, while the primary outcome 

measure was the use of life sustaining measures (cardiopulmonary 

resuscitation, mechanical ventilation, reintubation, 

hemodialysis, total parenteral nutrition, tracheostomy, and use 

of vasopressors). Patient and hospital demographic information 

as well as patient comorbidities were controlled for in multivariate 

analysis. Results: There were 6,600 patients with cirrhosis 

of the liver, age 40 or greater, who died in the hospital. Of 

these, 4,178 had at least one form of hepatic decompensation 

and were then included in to our analysis. Comorbidity 

indexes across race were not significantly different. On univariate 

analysis, black and Hispanic race had significantly 

more frequent use of life sustaining measures (69% and 65%, 

respectively) compared to white race (58%, p


Disclosures: 

Vincent Di Martino - Board Membership: Merk schering Plough France, Merk 

schering Plough France, Merk schering Plough France, Merk schering Plough 

France; Speaking and Teaching: Roche France, Gilead France, Roche France, 

Gilead France, Roche France, Gilead France, Roche France, Gilead France 

The following authors have nothing to disclose: Armand Garioud, Jean françois 

D. Cadranel, Jean-Baptiste Nousbaum, Allaoua Smaïl, Hortensia Lison, Thierry 

Thevenot, Mourad Medmoun, Djamel Belloula, Firouzeh Kazerouni, Khaled 

Hadj-Nacer, Alain Cazier 

Table 1. Predictive parameters, risk score and estimated probability 

of survival. 

555 

Predicting survival in liver abscess: a risk score based 

on 8,423 patients hospitalized with liver abscess, Thailand. 

Kittiyod Poovorawan 1 , Wirichada Pan-ngum 2 , Ngamphol 

Soonthornworasiri 2 , Chatporn Kittitrakul 1 , Polrat Wilairatana 1 , 

Sombat Treeprasertsuk 3 , Bubpha Kitsahawong 4 , Kamthorn Phaosawasdi 

4 ; 1 Department of Clinical Tropical Medicine, Faculty 

of Tropical Medicine, Mahidol University, Bangkok, Thailand; 

2 Department of Tropical Hygiene, Faculty of Tropical Medicine, 

Mahidol University, Bangkok, Thailand; 3 Department of Medicine, 

Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand; 

4 Vichaiyut hospital and Medical Center, Bangkok, Thailand 

We aimed to establish a risk score for predicting 60-day survival 

in patients hospitalized with liver abscess, using Nationwide 

Hospital Admission Data from 2009 to 2013 provided 

by the National Health Security Office (NHSO), Thailand. All 

patients with a primary diagnosis of pyogenic liver abscess and 

amoebic liver abscess (ICD10-K750, A064) were included. 

Baseline characteristics, comorbidities, hospital course, and 

survival were analyzed, and all-causes mortality data were 

retrieved from national death registration. A total of 11,296 

admissions comprised of 8,423 patients from 844 hospitals 

across Thailand were eligible for analysis. The mean age was 

52±17 years and 66.1% of patients were male. Median length 

of hospitalization was 8 days (IQR 4 to 13 days), and the 

mean cost of hospitalization was 846±1,574 USD. Overall 

in-hospital mortality rate was 2.8%, and overall long-term survival 

was significantly higher among patients with amoebic 

liver abscess compared to pyogenic liver abscess (HR 0.54; 

95% CI, 0.42-0.68; p


Disclosures: 

The following authors have nothing to disclose: Michelle T. Jesse, Reema Habra, 

Steven Mekaru, Eric Goldstein, Anne Eshelman, Marwan S. Abouljoud 

557 

Palliative Care Services are Underutilized in Liver Transplant 

Candidates 

Priya Kathpalia 1 , Alexander Smith 2 , Jennifer C. Lai 1 ; 1 Division of 

Gastroenterology and Hepatology, University of California, San 

Francisco, San Francisco, CA; 2 Division of Geriatrics, University of 

California, San Francisco, San Francisco, CA 

Background: Patients with end-stage liver disease (ESLD) 

experience high symptom burden and 50% risk of death at 

5 years from progressive liver disease without liver transplant 

(LT). These patients represent a population with high needs 

for palliative care. Utilization practices of palliative care of LT 

candidates are not well characterized. Methods: All adult LT 

candidates who died or were delisted from the wait-list at a 

single high volume LT center from 2013-14 were evaluated. 

Excluded were those who were delisted for noncompliance 

or were transplanted at another center. Reasons for palliative 

care consultation were obtained from review of electronic 

health records. Logistic regression assessed factors associated 

with palliative care consultation in these patients. Results: Of 

683 patients listed for LT in 2013-14, 107 (16%) ultimately 

died (n=62) or were delisted for being too sick for LT (n=45): 

median age was 58 years, 34% were female, 53% were white, 

61% had Child Pugh Class C cirrhosis, and 52% had hepatocellular 

carcinoma. Among these 107 patients who died/were 

delisted, 17% (n=18) received a palliative care consult, 89% 

(16/18) of which occurred in the inpatient setting. The median 

(interquartile range [IQR]) number of days from palliative care 

consultation to death was 4 (1-11 days), with 50% of the consultations 

occurring within 3 days of death. Reasons for palliative 

care consultation were: assistance with transition to comfort 

care (78%), goals of care without transition to comfort care 

(11%) and symptom management (11%). Even among the 26 

patients delisted for advanced HCC, only 12% were referred 

to palliative care. In univariable analysis, age (OR 0.92; 95% 

CI 0.87-0.98; p


analyzed a large commercial insurance provider database to 

investigate the epidemiology, the pattern of treatment, and 

survival in patients with HCC. Methods: This is a retrospective 

matched case-control analysis estimating the epidemiology of 

HCC as well as clinical characteristics, treatment patterns and 

survival in enrollees with HCC in Health Care Services Corporation 

(HCSC), a commercial insurer providing health-care coverage 

in 5 states, from September 2008-August 2013. HCC 

was defined as ICD-9 code 155.0 after the first six months 

of enrollment. Cirrhosis was defined by ICD-9 codes 571.2, 

571.5, and 571.6. Treatment was grouped as following: surgical 

resection (CPT: 47120, 47122, 47125, 47130, 47712), 

ablation (CPT: s 47370, 47371, 47379, 47380, 47381, 

47382), TACE (CPT: 36247, 36248, 36246, 75726, 75774, 

37243, with code 96420) TARE (CPT: 36247, 36248, 36246, 

75726, 75774, 37243, with code 79445). Sorafenib use 

was defined as:ICD9 155.0 and a claim for sorafenib. Liver 

transplant was defined as ICD-9: v42.7 AND CPT 47135 or 

47136. Results: During the study period, there were 3465 

with an ICD-9 code 155.0 among 3,478,093 enrollees yielding 

a prevalence of HCC of 0.09%. Among patients with cirrhosis, 

7.95% had a diagnosis of HCC. When compared to 

those with cirrhosis without HCC, those with HCC were older 

(58.1 vs 55.5, p


≥1.45 were counseled by the HIV clinic and referred to the 

Division of Gastroenterology and Hepatology for liver liver 

fibrosis assessment and evaluation for HCV therapy, as reimbursement 

of second-generation direct-acting antivirals is 

restricted to patients with ADVFIB (liver stiffness >9.5kPa/ 

METAVIR F3/F4) in Austria. Results: Among 1348 HIV-positive 

patients, 31%(414/1348) of patients were HCV-antibody 

positive and 17%(229/1348) patients had detectable serum 

HCV-RNA. The majority of HIV/HCV (62%) had HCV-genotype 

1 (subtype 1a:75%/1b:25%), while HCV-genotypes 2, 

3, and 4 were observed in 1%, 28%, and 9% of patients. 

One hundred thirteen HIV/HCV with a FIB-4 index ≥1.45 

were counseled by the HIV clinic. Among these, 58%(66/113) 

underwent liver fibrosis assessment either by transient elastography 

(TE) or biopsy. ADVFIB was confirmed in 50% (33/66) 

and IFN-free treatments were initiated in 17 of these patients 

(SOF/RBV:12%[2/17]; SOF/DCV:76%[13/17]; SOF/ 

LDV:12%[2/17]). Importantly, among patients with a FIB-4 

index ≥1.45 in whom liver fibrosis was not assessed by either 

TE or biopsy (42%[47/113]), 13 patients had a FIB-4 index 

>3.45, which is diagnostic for ADVFIB. None of these patients 

received HCV therapy. Another 9 patients with a FIB-4 index 

576,000 

patients receiving AMA testing between 1/10 - 12/13 in 

a large US commercial laboratory database. Patients were 

considered AMA+ if antibody titre > 1:20 or M2 antibody > 

25.0 U. We examined patient demographics, site of testing 

& testing history. Results: 16,492 patients tested AMA+ and 

had at least 1 alkaline phosphatase (ALP) test over the 4 year 

period; 6,107 had an ALP >120 U/l that period and could be 

classified as probable PBC. 53% of initial AMA+ tests were 

conducted in a hospital setting. The specialty of the ordering 

outpatient physician was gastroenterologists (18%), primary 

care (14% including NP/PAs) and Rheumatologists (2%). A significant 

minority of 1st AMA+ tests were male (19%). The age 

at 1st AMA+ tests: 

65 (40%). 55% of patients had >3 ALP tests before 1st positive 

AMA; 67% had at least one ALP > 115 U/l before 1st AMA+ 

test, and 40% had >2 tests with ALP > 115. Conclusion: The 

63% of AMA+ patients without elevated ALP (ie, probably 

PBC) is similar to that reported in AASLD guidelines. Most PBC 

patients’ 1st AMA+ test was in a hospital setting, suggesting 

that they are being diagnosed as the result of standard screening 

through other emergent conditions, not as part of regular 

care. Given that early stage PBC is asymptomatic, this may 

not be unexpected, except that 67% of patients had > 1 and 

40% > 2 ALP test results > 115 U/l before 1st AMA+ test. 

This suggests that outpatient physicians may not be following 

up on elevated ALP with AMA testing. The distribution of age 

is similar to prevalence studies, but the proportion of men testing 

AMA+ is more than double what was expected. Other 

studies suggest that men are diagnosed at a later PBC stage 

than women, and have a higher mortality rate, thus a higher 

proportion of incident men would ultimately produce lower 

prevalence. Research on healthcare resource use shows that 

in the US, men are less likely to use primary care, and instead 

receive care through emergency rooms, which may explain the 

high rate of hospital testing. 

Disclosures: 







The following authors have nothing to disclose: Keith D. Lindor 

563 

Worse Outcomes Among Uninsured Patients with Cirrhosis 

– Towards Understanding the Obstacles to Hepatology 

Care 

Daniela Ladner 1 , Kathryn Skibba 1 , Kathryn Jackson 1 , Niloufar 

Safaeinili 1 , David S. Goldberg 2 , Lisa B. VanWagner 1 , Satyender 

Goel 1 , Abel Kho 1 , Amna Daud 1 , Nazanin Salehitezangi 1 , Anton 

I. Skaro 1 ; 1 Northwestern University, Chicago, IL; 2 University of 

Pennsylvania, Philadelphia, PA 

Background: Cirrhosis of the liver is a leading cause of morbidity 

and mortality in the US and worldwide. Advanced hepatology 

care and transplantation can improve survival and quality 

of life. However, discrete obstacles to access to appropriate 

liver-specific care exist. Due to an absence of epidemiologic 

data collected systematically on the cirrhotic patient population 

the nature and magnitude of the problem remains poorly characterized. 

Longitudinal measurement of MELD is an adequate 

surrogate of access to care. Therefore, we examined outcomes


of patients with cirrhosis identified within the greater Metropolitan 

Chicago area by query of the HealthLNK database. Methods: 

HealthLNK captures over 2 million patients treated within 

6 diverse healthcare integrated delivery networks in the greater 

Chicago area (Northwestern Medicine, Univ of Chicago, Rush 

Univ MC, University of Illinois of Chicago MC, and Loyola Univ 

MC, and Cook County Health Systems) between 2006-2012. 

Cirrhosis was defined by ICD-9 codes 571.2 OR 571.5 OR 

571.6 OR Fib-4 score > 3.25. Demographics, and co-morbidities 

assessed by ICD-9 codes, CPT codes, medications and laboratory 

values were collected. Cirrhotic patients were stratified 

into groups according to whether or not the MELD score was 

measured. Comparisons by group were tested using t-tests or 

chi-squared tests for continuous and non-continuous variables 

respectively. Results: 24,185 cirrhotic patients were identified 

and 11,481 (47%) had INR, bilirubin, and creatinine biochemistries 

drawn at one or more points in time. There were no 

differences in age or sex among those patients with no labs 

available to calculate a MELD score versus those with labs to 

calculate a MELD score (p2) were assimilated into 

a screening tool. The tool was retrospectively applied to all 

patients admitted to the Bristol Royal Infirmary with a diagnosis 

of cirrhosis over 90 consecutive days from 1st July 2013 

(n=54). Mortality 1 year post initial admission was calculated. 

Results Of 54 index cases 7 were not analysed due to incompleteness 

of data or loss to follow up. On the basis of optimum 

sensitivity and specificity for death at 1 year a score of 3 or 

more was considered a threshold for identifying likely poor 

prognosis (table). Conclusions The tool has been trialed and 

audited over the past year. Patients who screen positive (≥ 

3 criteria) are discussed at a weekly hepatology multidisciplinary 

meeting (MDM). Assuming MDM agreement consultant 

led patient discussion, a poor prognosis letter to the GP, and a 

palliative medicine referral are triggered. Communication skills 

training has been delivered to consultant and junior staff by 

the palliative medicine team. As deaths from liver disease continue 

to increase, identifying patients who stand to benefit from 

advanced care planning and timely palliative care intervention 

will become increasingly important. 

Accuracy of tool at varying ‘thresholds’ for predicting mortality at 

1 year 

Disclosures: 

The following authors have nothing to disclose: Benjamin E. Hudson, Kelly Ameneshoa, 

Peter Collins, Andrew J. Portal, Fiona H. Gordon, Julia Verne, Anne 

McCune 

565 

Distance to Transplant Center and Waiting List Outcomes 

Jordan Voss, Adam McCann, Sean C. Kumer, Timothy Schmitt, 

Richard Gilroy; Kansas University Medical Center, Kansas City, KS 

Objective: To analyze the impact of a patient’s distance from 

a transplant center on liver transplant waiting list outcomes. 

Methods: All patients waitlisted at a single transplant center 

from January, 2010 to May, 2014 were evaluated for waitlist 

outcome, distance to transplant center and model for end-stage 

liver disease (MELD) score at time of listing, transplant, and 

death. Distance to transplant center was evaluated both as a 

continuous variable and categorically for distances of 25, 50, 

100, and 200 miles. Logistic regression was used to evaluate 

the effect of distance on likelihood of death on the waiting list. 

Results: 617 patients were waitlisted during the study period. In 

univariate analysis distance was very weakly associated with a 

lower MELD at both listing and death: R=-0.086, p=0.033 and 

R=-0.181, p=0.036, respectively. Logistic regression revealed 

that distance does not predict death on the waiting list (OR 

1.000, p=0.851). Median test and Mann-Whitney U Test indicated 

that MELD scores at listing, transplant, and death are 

not significantly different among patients living within 25, 50, 

100, and 200 miles of the transplant center compared to those 

living outside the specified distances. Conclusions: A patient’s 

distance from the liver transplant center does not appear to 

impact MELD score at the time of listing and once listed does 

not impact MELD at transplant/death or likelihood of death on 

the waiting list. How socioeconomic and racial/ethnic variables 

impact this warrants further consideration given the differ-


ence in these variables between rural and urban populations, 

and is being evaluated in subsequent analyses. 

Disclosures: 

Richard Gilroy - Speaking and Teaching: Salix, NPS, Gilead, AbbVie, Novartis 

The following authors have nothing to disclose: Jordan Voss, Adam McCann, 

Sean C. Kumer, Timothy Schmitt 

566 

Deleterious effects of cirrhosis on outcomes after firearm 

injury 

Sentia Iriana 1 , Elliot B. Tapper 2 , Vinay Sundaram 1 ; 1 Medicine, 

Cedars-Sinai Medical Center, Los Angeles, CA; 2 Medicine, Beth 

Israel Deaconess Medical Center, Boston, MA 

Aims: Cirrhotic patients have poor outcomes from traumatic 

injury. The impact of firearm injury in this population is unexplored. 

The aim of this study was to assess the prevalence and 

outcomes related to firearm injury related hospitalization in cirrhosis 

patients. Methods: We analyzed the Nationwide Inpatient 

Sample, 2009-2011. Patients were identified by having a 

primary or secondary discharge diagnosis associated with firearm 

injury. We utilized multivariable logistic regression models 

for the outcome of inpatient mortality. Deyo modification of the 

Charlson index was used to account for non-hepatic comorbidity. 

Results: A total of 348,823 discharge records were studied, 

of which 317,592 had cirrhosis alone, 26,044 had firearm 

injury without cirrhosis, and 2,730 had cirrhosis with firearm 

injury. Among cirrhotics with firearm injury, 910 (33.3%) were 

accidental and 1820 (66.7%) were assault related. Cirrhotics 

with firearm injury were younger compared to cirrhotics without 

firearm injury (54.9 vs 58.7 years, p


lyzed using Chi2 test for trend and linear regression analysis. 

Results Among 34,308 HIV-infected patients enrolled between 

2000 and 2012, 5,562 were HCV coinfected. HCV prevalence 

declined from 28% to 15% during the study period 

(p


outcomes in a multi-center cirrhotic cohort. Methods: NACSELD 

(North American Consortium for End-Stage Liver Disease) 

enrolls hospitalized cirrhotics from 16 centers. Data collected 

are admission/cirrhosis details, inpatient details & 30-day mortality. 

Details & outcomes of pts given (Alb) /not given albumin 

(no Alb) and regional variations were compared. Within 

Alb, evidence-based indications (AKI, SBP, paracentesis) were 

compared to others. Results: 988 cirrhotics (64% men, MELD 

19, 43%alcohol) were included. 597(60%) pts received a 

280±267 gm of albumin. Alb pts had worse cirrhosis severity 

but similar demographics as non-alb(Table).Albumin use: The 

majority(76%) was evidence-based (47%AKI, 34%paracentesis 

& 14%SBP). Remaining 24% was for hyponatremia (12%), 

non-SBP infection (6%) & anasarca (6%). This resulted in AKI 

resolution in 28%, reduced anasarca in 22%, improved Na in 

12% & allowed paracentesis in 30% of pts. 4% had adverse 

events (3% fluid overload & 1% pulmonary edema) Regional 

variation: 4 regions: 3 US (1: North, 2: South, 3: Southwest) 

& Canada (region 4) were studied. Region 3 had the highest 

albumin use (69%) with lowest evidence-based use (57%) compared 

to Region 2 that used the least(45%,p


The following authors have nothing to disclose: Luca Fabris, Stefano Okolicsanyi, 

Matteo Rota, Paolo Cortesi, Luca S Belli, Stefano Fagiuoli, Luciana Scalone, 

Giancarlo Cesana, Mario Strazzabosco 

572 

Knowledge about hepatitis B transmission risks among 

health professionals in Tanzania 

Jose Debes 1 , Johnstone Kayandabila 2 , Hope Pogemiller 1 ; 1 University 

of MInnesota, Minneapolis, MN; 2 Arusha Lutheran Medical 

Centre, Arusha, United Republic of Tanzania 

Introduction: It is well known that hospital workers are at 

increased risk for contracting hepatitis B virus (HBV). This risk 

it is increased in settings of high HBV seroprevalence, such as 

sub-Saharan Africa. Despite this, it is unclear whether healthcare 

providers in developing countries are aware of the transmission 

risks and adhere to universal precaution strategies. 

In this study we aimed to evaluate the knowledge and understanding 

of HBV among health-care workers in hospitals in 

Tanzania. Methods: We provided a HBV survey to staff in 2 

hospitals in northern Tanzania. The survey consisted of nine 

multiple-choice questions that inquired about understanding 

of HBV serostatus, vaccination and risks of transmission of 

HBV. The survey was written in English and Swahili (the local 

language) and distributed among medical and non-medical 

staff (laboratory technicians, students, etc). Multivariate analyses 

were performed using Fishers exact test and Chi-square. 

Results: We received voluntary participation from 114 subjects 

in our survey. The mean age of the participants was 33 

y/o, 67% were females, and 61% responded to the survey in 

English. Ninety one percent of subjects had no knowledge of 

their HBV surface antigen status, and 89% indicated they never 

received a vaccine for HBV, with lack of knowledge about 

the vaccine being the most common reason (34%). Seventy 

percent of participants new about HBV complications and 60% 

responded correctly inquires about transmission routes. There 

was a significant difference in knowledge of HBV serostatus 

and vaccination between participants with a medical background 

(consultants, interns, etc) and others, p=0.01 and 

p-0.001 respectively. However, only 33% of consultants knew 

about their HBV serostatus or were vaccinated for it. There 

was no significant difference between knowing HBV transmission 

route or in-hospital risk and whether the survey was 

answered in English or local language, or hospital position of 

the responder. Conclusions: Our study shows a surprisingly low 

knowledge of HBV serostauts and vaccination status among 

hospital workers in Tanzania. Studies are needed in other parts 

of sub-Saharan Africa to further understand this gap in knowledge, 

and HBV awareness programs should be promoted. 

Disclosures: 

The following authors have nothing to disclose: Jose Debes, Johnstone Kayandabila, 

Hope Pogemiller 

573 

Hepatitis C Virus Sustained Virologic Response Rates 

among Patients Treated in Primary Care Settings in New 

York State 

Colleen Flanigan, Jason Pendergast; New York State Department 

of Health, Albany, NY 

Background Hepatitis C virus (HCV) treatment has been rapidly 

changing over the last few years. With the arrival of direct 

acting anti-viral therapies, HCV treatment is now highly effective. 

As a result of these more effective treatments, the demand 

for HCV treatment is increasing, adding strain to an already 

limited number of specialists available to treat HCV. The newer 

treatments are less complex, more tolerable and treatment 

durations can be as short as eight weeks, thus, allowing for 

primary care providers (PCPs) to treat HCV. To expand access 

to HCV care and treatment in New York State (NYS), the NYS 

Department of Health provides funding to 13 primary care sites 

to integrate HCV care and treatment. These primary care sites 

include: community health centers, hospital-based clinics, a 

methadone maintenance treatment program and a residential 

drug treatment program. At each of these settings, a PCP and 

a multidisciplinary team manages and treats persons infected 

with HCV. Each PCP is an experienced HCV provider with the 

knowledge and skills to safely and effectively manage and 

treat HCV. Each PCP has an agreement with a liver specialist 

for consultation. The purpose of this study was to evaluate 

HCV treatment sustained virologic response (SVR) rates 

among patients receiving HCV treatment in a primary care 

setting. Methods This analysis looks at HCV SVR rates among 

HCV monoinfected and HIV/HCV coinfected patients enrolled 

at 13 primary care sites across NYS. All primary care sites 

report client level data using the AIDS Institute Reporting System 

(AIRS). AIRS is a comprehensive client and service/encounter 

reporting application. AIRS also collects HCV treatment type 

and treatment outcome (SVR). Results From October 1, 2010 

through April 30, 2015, 2,447 HCV infected patients were 

served by the 13 primary care sites. Of these patients, 816 

(33.3%) initiated HCV treatment and of these patients 743 

(91.1%) had final treatment outcomes available for analysis. 

The overall SVR rate among these patients was 61.5%; 70.1% 

among HCV monoinfected and 50.8% among HIV/HCV coinfected 

patients. SVR rates ranged from 49.3% among those 

treated with the combination pegylated interferon and ribavirin 

to as high as 96.6% among those treated with ledipasvir-sofosbuvir. 

Conclusions These findings illustrate that HCV treatment 

SVR rates are just as good in primary care settings as they are 

in specialty settings. Therefore, integrating HCV treatment into 

primary care can help increase capacity for HCV treatment 

and allow more people to be cured of HCV. 

Disclosures: 

The following authors have nothing to disclose: Colleen Flanigan, Jason Pendergast 

574 

External Validity of Trials in Non-Alcoholic Fatty Liver 

Disease: Systematic Review of Randomised Controlled 

Trials and Non-Randomised studies 

Richard Parker 1 , James Hodson 2 , Ian A. Rowe 1 ; 1 Centre for Liver 

Research, University of Birmingham, Birmingham, United Kingdom; 

2 University Hospitals Birmingham NHS Foundation Trust, Birmingham, 


Introduction Trials with good external validity allow clinicians 

and patients to have confidence in new treatments. Individuals 

in clinical trials are often highly selected, in part to remove 

confounding factors, but this can introduce bias and limit the 

external validity of a trial. In this study, we surveyed observational 

studies and randomised controlled trials (RCT) to gauge 

the external validity of trials in non-alcoholic fatty liver disease 

(NAFLD). Methods Structured literature searches were undertaken 

for observational studies (ObS) and RCT in NAFLD. Data 

were extracted from included studies. Participant characteristics 

were compared between observational studies and RCT, and 

between subsets of observational studies describing with biopsy-proven 

non-alcoholic steatohepatitis (NASH) or advanced 

fibrosis (>F2). For age and BMI, weighted means were calculated 

and compared with t-tests, for prevalence of Diabetes mellitus 

(DM) and gender, Chi-squared was used for comparisons.


Results 112 studies were included: 47 RCT and 65 ObS. Sixteen 

observational studies described characteristics of patients 

with NASH and 31 described characteristics of patients with 

advanced fibrosis. RCT participants differed from individuals in 

ObS with regard to age, BMI, prevalence of DM, and gender 

(p


disseminate knowledge on new HCV treatments are needed to 

decrease the burden of HCV infection. 

577 

Interventions to optimize retention in the chronic viral 

hepatitis screening, care and treatment cascade: a systematic 

review 

Kali Zhou 4 , Nick Walsh 1 , Thomas Fitzpatrick 2 , Ji Young Kim 1 , 

Ying-Ru Lo 1 , Joseph D. Tucker 3 ; 1 World Health Organization Western 

Pacific Region, Manila, Philippines; 2 University of Washington, 

Seattle, WA; 3 University of North Carolina-Chapel Hill, Chapel 

Hill, NC; 4 Gastroenterology, UCSF, San Francisco, CA 

BACKGROUND: Global mortality burden of viral hepatitis is 

estimated at 1.45 million and now surpasses that of HIV. Recent 

advances in hepatitis B (HBV) and hepatitis C (HCV) therapeutics 

provide the opportunity to effectively treat HBV and cure 

chronic HCV, thereby reducing hepatitis related mortality. To 

support countries in the delivery of hepatitis programs, we carried 

out a systematic review of interventions that may increase 

uptake and retention in the hepatitis screening, care and treatment 

service cascade. METHODS: We searched Pubmed, 

EMBASE, the WHO library, Clinicaltrials.gov, International 

Clinical Trials Registry Platform, Psychinfo and Cinahl. Interventional 

studies with a comparator arm targeting an aspect of the 

chronic hepatitis treatment cascade were included. The study 

was registered in PROSPERO (42014015094) and carried out 

according to PRISMA guidelines. Results from studies with similar 

outcomes were pooled using RevMan. RESULTS: We identified 

7,569 citations and included 56 in the review. All studies 

were from high-income settings. Self-reported HBV testing rates 

were higher among recipients of community based HBV-specific 

educational interventions compared to non-specific health 

interventions (RR=3.97, CI95 3.02-5.22). For HCV screening, 

facility-based interventions targeting providers (n=4) (HCV-specific 

education, electronic health record prompts) increased 

HCV testing rates (RR=3.44, CI95 3.24-3.65) compared to 

standard of care; interventions targeting patients (HCV-specific 

patient education) (n=4) also increased testing rates (RR=1.43, 

CI95 1.29-1.59) compared to standard of care. Interventional 

studies (n=11) to boost adherence to HCV treatment (patient 

support program, adherence literacy, integrated care program) 

increased treatment completion rates (RR=1.10, CI95 1.04- 

1.16)) compared to standard of care. Finally, when adherence 

was measured by sustained virologic response (SVR), 

adherence focused interventions (n=17) increased the likelihood 

of achieving SVR (RR=1.20, CI95 1.13-1.28) compared 

with standard of care. CONCLUSION: There was a lack of 

data on community-based interventions to increase HCV testing 

uptake, and no data on interventions to improve hepatitis 

B treatment uptake and adherence. These areas require further 

research. Efforts are needed to translate these findings to 

increase chronic viral hepatitis screening uptake and optimize 

country programmatic efforts in treatment. 

Disclosures: 

The following authors have nothing to disclose: Kali Zhou, Nick Walsh, Thomas 

Fitzpatrick, Ji Young Kim, Ying-Ru Lo, Joseph D. Tucker 

Disclosures: 

Anna S. Lok - Advisory Committees or Review Panels: Gilead, MYR, Tekmira; 

Consulting: GSK, Merck; Grant/Research Support: AbbVie, BMS, Gilead, Idenix 

The following authors have nothing to disclose: Mary Thomson, Monica Konerman 

578 

Upper Limit Normal of Alanine Aminotransferase for the 

Indian Population 

Jack Masur 1 , Shyam Kottilil 1 , Eleanor Wilson 2 , Samir R. Shah 3 ; 

1 Institute of Human Virology, University of Maryland School of 

Medicine, Baltimore, MD; 2 Critical Care Medicine Department, 

National Institutes of Health, Bethesda, MD; 3 Global Hospitals, 

Mumbai, India 

Introduction AASLD recently lowered the Upper Limit Normal 

(ULN) of alanine aminotransferase (ALT) level to 30 U/L in 

males and 19 U/L in females. Studies have found that relying 

on pre-existing normal ranges for ALT measurement in patients 

with chronic hepatitis C virus (HCV) infection was insensitive 

at identifying patients with minimal to mild necroinflammatory 

activity. ALT values have been shown to differ by ethnicity; 

as well as by metabolic factors such as BMI, glucose, cholesterol, 

and hyperlipidemia; and, by infection from chronic 

hepatitis viruses. The existing ULN level in India is 45 U/L, 

irrespective of sex. India currently has an HBV prevalence of 

3.7% and HCV prevalence of 1%, and many infected persons 

are unaware of their chronic carrier status. Additionally, 

non-alcoholic fatty liver disease (NAFLD) is an epidemic in 

India. Increased sensitivity of testing might allow for prevention 

and earlier intervention. The purpose of this study was to test 

whether the currently defined ULN of ALT thresholds in India 

underestimate the number of patients at risk for chronic liver 

disease in the country. Methods We conducted a retrospective 

study of patient laboratory data and vital sign measurements 

collected from the Metropolis Laboratory in Mumbai, India over 

a two-year period. Patients were analyzed based on preset 

health parameters that were used to determine inclusion or 

exclusion from the “control” cohort. The parameters required 

for inclusion in the “control” cohort included: BMI ≤23 kg/ 

m 2 ; normal blood pressure of 120/80 mm Hg; total cholesterol 


Disclosures: 

The following authors have nothing to disclose: Jack Masur, Shyam Kottilil, Eleanor 

Wilson, Samir R. Shah 

579 

Successful Implementation of a Community-based 

Patient Navigation Program to Increase Screening and 

Linkage-to-care in High-Risk Patients with Chronic Hepatitis 

B Infection 

Jennifer M. Newton 1 , Matt Johnson 2 , Sharon Song 2 , Helen S. Te 1 , 

Edwin Chandrasekar 2 , Karen E. Kim 1 ; 1 Medicine, University of 

Chicago Medicine, Chicago, IL; 2 Asian Health Coalition, Chicago, 

IL 

Background: Chronic hepatitis B virus infection (CHB) is estimated 

to affect up to 2.2 million people in the United States, 

and disproportionately affects foreign-born Asian and African 

immigrants. CHB carries a high morbidity and mortality 

due to undertreatment, which largely reflects lack of awareness 

of infection and poor linkage to care. Purpose: In this 

intervention study, we have implemented a community-based 

patient navigation program. Utilizing community-health workers 

(CHWs) and patient navigators (PNs), we aim to increase 

patient awareness of infection and improve linkage to care 

for foreign-born Asian, Pacific Islander and African patients 

with CHB. Methods: While CHWs provide culturally competent 

education, PNs are essential for care coordination. To improve 

community linkage to care, we developed a co-curriculum to 

enhance care coordination and community linkage for hepatitis 

B surface antigen (HBsAg) positive patients. To strengthen 

their partnerships, CHWs and HPNs jointly attend site visits 

and training sessions for cultural competency, CHB education, 

navigating the healthcare system and community awareness. 

Results: In the first 6 months of the intervention, 46 (4.7%) of the 

974 individuals screened were HBsAg positive. Asian (674, 

69.2%) and Black/African (224, 23%) patients accounted for 

the largest ethnic groups screened, while Iraq (228, 23.4%) 

and Myanmar (147, 15.1%) represent the largest screenings 

by country of origin. 247 patients received all 3 HBV serologic 

tests (surface antigen, surface antibody, and core antibody). 

Of these patients, 111 (44.9%) were previously vaccinated, 

60 (24.3%) were susceptible to infection, and 40 (16.2%) had 

resolved infection. 891 (91.5%) of the 974 patients screened 

received test results and counseling, including all 46 (100%) 

patients with active infection. Among patients with active 

HBV infection, 37 (80.4%) have been linked to care, with 31 

(67.4%) seeking care through specialty clinics and 6 (13%) 

through primary care. 3 (6.5%) patients moved out of state, 

and 6 (13%) have been lost to follow-up. Conclusions: To our 

knowledge, this is the first community-based hepatitis B patient 

navigation program to be instituted in the country. This intervention 

has been successful in linking high-risk Asian and African 

patients to healthcare for CHB, and is a promising model that 

may be implemented nationally to decrease the unnecessary 

morbidity and mortality associated with CHB. 

Disclosures: 



The following authors have nothing to disclose: Jennifer M. Newton, Matt Johnson, 

Sharon Song, Edwin Chandrasekar, Karen E. Kim 

580 

Novel Algorithm to Predict Hospital Readmission in 

Patients with Cirrhosis Utilizing Machine Learning Classifiers 

Spencer L. James 1 , Shawn Shah 2 , Zilla H. Hussain 2 , Neil Volk 2 , 

Joseph Shatzel 2 , Rolland Dickson 2 ; 1 Medical School, Geisel 

School of Medicine at Dartmouth, Lebanon, NH; 2 Department of 

Medicine, Dartmouth-Hitchcock Medical Center, Lebanon, NH 

BACKGROUND: Patients with cirrhosis have high rates of hospital 

readmission. Current models used to predict readmission 

in cirrhotic patients are based on limited data. The aims of this 

study are to use machine learning classifiers (MLC), a novel 

discipline of computer science focused on predicting outcomes 

from complex datasets, to develop a model which can accurately 

predict which cirrhotic patients will be readmitted after 

discharge. METHODS: We retrospectively reviewed patients 

with a primary diagnosis of cirrhosis admitted to a tertiary 

care center over a 21-month period (April 2011 to December 

2012). Using admission data, we applied various MLC 

and a deep learning technique to develop a predictive model 

for readmission in cirrhotics. The MLC utilized features of the 

patient’s EHR including age, sex, serum sodium, platelet count, 

hemoglobin, hepatic biomarkers, albumin, international normalized 

ratio, MELD score, use of rifaximin or lactulose, use of 

diuretics, use of a beta-blocker, prior TIPS, presence of hepatocellular 

carcinoma, etiology of cirrhosis and socioeconomic 

factors (such as income and insurance). We then tested the 

model’s ability to predict the likelihood of readmission among 

cirrhotic patients in the database. RESULTS: During the study 

period, 317 unique patients accounting for a total of 498 

admissions were included. The average patient age was 57.9 

years (range of 14 to 88), and 58% were male. The mean 

number of admissions per patient was 2.4 (range of 1 to 9), 

with an average time to readmission of 88 days (range of 1 

to 411 days). From this sample, we trained machine learning 

classifiers on a randomly selected 80% of the data, and 

then tested the predictive accuracy on the remaining 20%. 

This train/test process was repeated 100 times. The mean 

accuracy to correctly identify hospital readmission varied by 

classifier, with random forest classification correctly predicting 

patient readmission 70% of the time on average (range of 

53% to 80%). Random forest demonstrated a mean sensitivity 

and specificity of 50% and 85%, respectively, with a PPV and 

NPV of 70% and 69%, respectively, in the native distribution 

of the dataset. CONCLUSION: Machine learning classifiers 

were capable of predicting hospital readmission for patients 

with cirrhosis nearly 70% of the time. As a decision support 

tool, machine learning is promising to help risk stratify patients 

admitted with cirrhosis. With further validation, such classifiers 

could be incorporated into EHR systems to help identify highrisk 

patients with cirrhosis who are likely to be readmitted, and 

prompt appropriate multidisciplinary intervention. 

Disclosures: 

The following authors have nothing to disclose: Spencer L. James, Shawn Shah, 

Zilla H. Hussain, Neil Volk, Joseph Shatzel, Rolland Dickson


581 

Differentially altered gut microbial composition correlates 

with plasma metabolite alterations in patients of 

severe alcoholic hepatitis (SAH) and steroid response 

Shvetank Sharma, Jaswinder S. Maras, Sukanta Das, Shabir Hussain, 

Saggere M. Shasthry, Shiv K. Sarin; Institute of Liver & Biliary 

Sciences, New Delhi, India 

Purpose: Gut microbiota plays an important role in systemic 

metabolism and liver diseases. We hypothesized that gut 

microbiota and their metabolic pathways vary in SAH at baseline 

& on steroid therapy.Patients and Methods: From108 consecutive 

alcoholic liver disease patients, 60 with SAH [chronic 

alcohol use >5years, DF>32, liver histology suggestive of 

SAH, absence of known liver disease or h/o immunosuppressive 

drugs] were treated with prednisolone(40mg/day) 

for 28days. Response to steroids was assessed at day7 by 

Lille score [0.45=non-responder(NR)]. 

Stool samples were collected at baseline & day7 of therapy. 

250mg frozen samples were processed for DNA isolation & 

16S rRNA (V3-V4 region )analysed by NGS. Plasma was analysed 

for metabolites by MS/MS.Results: Total of 305 species 

from 135 genera were identified among six R & NR each. 

Klebsiella(32%), Enterococcus(24%) were identified in NR, 

while Bacteroides(26%),Lactobacilli(21%) were identified in R. 

Post-therapy, E.coli(45%) & Bifidobacterium(21%) were dominant 

in R &E.coli(40%) &Lactobacillus(26%) in NR.At baseline 

R & NR were categorized as type1 & type2 enterotypes. 

Post-therapy both groups demonstrated type3 composition.34 

metabolic pathways were common between all the treatment 

groups & time-points.Ferredoxin oxidoreducatse,& LPS biosynthesis, 

sulphate transport, Sec-system pathways were uniquely 

present in R0 &NR0 respectively, suggesting increase in the 

secretory phenotype in NR at baseline. Significant upregulation 

(1.5 fold, p


583 

Phenotypic and genotypic characterization of drug-induced 

liver injury (DILI) with autoimmune features 

C. Stephens 1 , A. Ortega 1 , I. Medina-Cáliz 1 , Mercedes Robles 

Diaz 1 , Agustin Castiella 2 , Pedro Otazua 3 , E. Zapata 2 , E.M. 

Gomez-Moreno 4 , MAngel López-Nevot 4 , Francisco Ruiz-Cabello 4 , 

German Soriano 5 , E. Roman 5,6 , Hacibe Hallal 7 , Jose María 

Moreno-Planas 8 , Martin Prieto 9 , M. I. Lucena 1 , Raul J. Andrade 1 ; 

1 Instituto de Investigación Biomédica de Málaga (IBIMA), Hospital 

Universitario Virgen de la Victoria, Universidad de Málaga, 

CIBERehd, Málaga, Spain; 2 Hospital Mendaro, Guipúzcoa, Spain; 

3 Hospital Mondragón, Guipúzcoa, Spain; 4 Instituto de Investigación 

Biosanitario de Granada, Hospital Universitario Virgen de 

las Nieves, Universidad de Granada, Granada, Spain; 5 Hospital 

de la Santa Creu i Sant Pau, Universitat Autònoma de Barcelona, 

CIBERehd, Barcelona, Spain; 6 Escola Universitària d’Infermeria 

EUI-Sant Pau, Universitat Autònoma de Barcelona, Barcelona, 

Spain; 7 Hospital Morales Meseguer, Murcia, Spain; 8 Complejo 

Hospitalario Universitario de Albacete, Albacete, Spain; 9 Hospital 

La Fe, CIBERehd, Valencia, Spain 

Background: Positive autoantibody (AAB) titres is a common 

presentation of autoimmune hepatitis (AIH), but can also occur 

in DILI. The underlying mechanism of selective AAB occurrence 

in DILI is unknown. Hence, we aimed to compare demographics, 

clinical parameters and HLA allele compositions in DILI 

with positive (+) and negative (-) AAB titres and AIH patients. 

Material and Methods: High resolution genotyping of HLA 

class I (A, B, C) and II (DRB1, DQB1) loci were performed 

on 207 DILI (drug-induced autoimmune hepatitis excluded) 

and 51 AIH patients and compared to 885 Spanish healthy 

controls. Results: Fifty-eight of the 207 DILI patients presented 

positive titres for at least one AAB (ANA 76%, ASMA 28%, 

AMA 9% or LKM-1 2%) during the DILI episode, while 149 

were negative for all four AABs. Comparing demographics 

and clinical parameters, AAB+ patients were found to be significantly 

older than AAB- patients (58 vs 50 years, p=0.019). 

Females predominated among the AIH (71%) compared to 

AAB+ (59%) and AAB- (47%), p=0.01. Significant differences 

in type of liver injury were present (p=0.0025), with cholestatic/mixed 

damage being more prevalent in AAB-, which was 

reflected in a higher onset ALP elevation in the same group 

(p=0.029). Furthermore, hypertension was significantly more 

frequent in AAB+ patients. Compared to controls, HLA alleles 

B*08:01 (45% vs 10%, pc=1.0E-12), C*07:01 (47% vs 24%, 

pc=0.006), DRB1*03:01(59% vs 26%, pc=2.0E-05) and 

DQB1*02:01 (57% vs 22%, pc=3.0E-07) were significantly 

more frequent in AIH patients. The frequency of HLA-A*01:01 

was increased in the same population, but did not reach significance 

after Bonferroni’s correction (33% vs 19%, p=0.02/ 

pc=0.37). There was a tendency for higher representation of 

DRB1*14:01 and DQB1*05:03 in DILI AAB+ compared to 

DILI AAB- (14% vs 4.7%, p=0.03; 14% vs 5.4%, p=0.06) and 

controls (14% vs 5.0%, p=0.007; 14% vs 5.4%, p=0.01). 

Conclusions: The AAB+ patients were found to be older and 

less prevalent to develop cholestatic/mixed injury compared 

to AAB- patients. The AIH patients were predominately female 

with hepatocellular injury and less likely to have underlying 

hypertensive conditions. The presence of HLA alleles B*08:01, 

C*07:01, DRB1*03:01 and DQB1*02:01 and possibly 

A*01:01 appear to enhance the risk of AIH in Caucasians with 

Spanish inheritance. These alleles form part of the conserved 

extended haplotype 8.1. However, haplotype formations in the 

study cohort are currently unknown. HLA alleles DRB1*14:01 

and DQB1*05:03 could potentially increase the risk of positive 

AAB (particularly ANA) in Spanish DILI patients. Funding: 

P10-CTS-6470, PI12/00378, AC-0073-2013, CIBERehd-ISCIII 

Disclosures: 

Martin Prieto - Advisory Committees or Review Panels: Bristol, Gilead 

The following authors have nothing to disclose: C. Stephens, A. Ortega, I. Medina-Cáliz, 

Mercedes Robles Diaz, Agustin Castiella, Pedro Otazua, E. Zapata, 

E.M. Gomez-Moreno, MAngel López-Nevot, Francisco Ruiz-Cabello, German 

Soriano, E. Roman, Hacibe Hallal, Jose María Moreno-Planas, M. I. Lucena, 

Raul J. Andrade 

584 

Effect of Branched Chain Amino Acids on Iron Transport 

in the Liver and Small Intestine of a Cirrhotic Rat Model 

Yoshinao Kobayashi 1,2 , Ryosuke Sugimoto 2 , Motoh Iwasa 2 , Yoshiyuki 

Takei 2 ; 1 Center for Physical and Mental Health, Mie University 

Graduate School of Medicine, Tsu, Japan; 2 Department of Gastroenterology 

and Hepatology, Mie University Graduate School of 

Medicine, Tsu, Japan 

BACKGROUND: Branched chain amino acids (BCAA) have 

been used as a supplemental therapy to improve malnutrition 

and event-free survival in patients with liver cirrhosis (LC). Iron 

is known to involve in inflammation and carcinogenesis of the 

liver through production of reactive oxygen spices (ROS). We 

have reported that BCAA reduce oxidative stress by diminished 

hepatic iron accumulation in a cirrhotic rat model. In order to 

clarify the effect of BCAA supplementation on iron metabolism 

of the liver and small intestine, we investigated effects of BCAA 

on hepatic iron accumulation and intestinal iron transporters 

in a rat model. Additionally, we investigated effects of BCAA 

on expression of intestinal iron transporters using CaCo2 cells. 

METHODS: Carbon tetrachloride (CCl 4 

) was administered to 

male Wistar rats. On the 5 th week, rats were randomly divided 

into 3 groups and further maintained for 16 weeks. Group 1 

(G1) was treated with basal diet as a control (n=8). Group 2 

was treated with non-heme iron diet (60mg/kg a day) (n=7). 

Group 3 was treated both with non-heme iron and BCAA 

mixture (10mg/kg a day) (n=7). On the 21 st week, liver and 

intestine tissues were collected. Expression of iron transporters, 

divalent metal transporter 1 (Dmt1) and ferroportin 1 (Fp1), 

was analyzed by a semi-quantitative RT-PCR and Western blotting. 

Furthermore, Caco2 cells were cultured for 48 hours with 

FeCl 3 

, deferoxamine (DFO) and/or BCAA. RESULTS: We have 

shown that BCAA-supplemented rats showed lower hepatic iron 

contents (P< 0.05), lower hepatic hepcidin mRNA expression 

(P< 0.001), lower serum hepcidin level (P= 0.05) and lower 

hepatic 8-hydroxyl-2’-deoxyguanosine (8-OHdG) production 

(P< 0.05), as compared with control rats. Protein expression 

of Dmt1 in the small intestine was reduced in G2 (vs. G1, P < 

0.05), which tended to be retrieved in G3 (vs. G2, P= 0.08). 

Protein expression of intestinal Fp1 tended to be reduced in G2 

(vs. G1, P=0.06). The reduced FP1 tended to be retrieved in 

G3 (vs. G2, P= 0.09). BCAA did not affect gene and protein 

expression of Dmt1 in Caco2 cells. Gene expression of Fp1 in 

these cells was not significantly changed by BCAA, whereas 

protein expression of Fp1 in the cell membrane fraction was 

reduced when cultured with BCAA. Among BCAA, leucine 

most strongly repressed FP1 protein expression in Caco2 cells. 

CONCLUSION: BCAA, especially leucine, can reduce intestinal 

Fp1 protein expression through post-transcriptional pathway, 

which was a possible mechanism for reducing intestinal iron 

absorption. BCAA and serum hepcidin level reduced by BCAA 

supplementation can modulate iron absorption from the intestine 

and hepatic iron accumulation. 

Disclosures: 

The following authors have nothing to disclose: Yoshinao Kobayashi, Ryosuke 

Sugimoto, Motoh Iwasa, Yoshiyuki Takei


585 

Persistent Generation of Inflammatory Mediators after 

Acetaminophen Overdose in Surviving and Non-Surviving 

Patients 

Benjamin Woolbright, Mitchell R. McGill, Matthew R. Sharpe, 

Hartmut Jaeschke; Pharmacology, Toxicology & Therapeutics, University 

of Kansas Medical Center, Kansas City, KS 

Acetaminophen (APAP) overdose is a leading cause of drug-induced 

liver failure. In both human overdose patients and the 

murine model, metabolic activation of acetaminophen leads to 

formation of protein adducts, mitochondrial dysfunction, necrosis 

and an acute inflammatory response. Considerable debate 

occurs over the role of the inflammatory response after APAP 

overdose, as to whether sterile inflammation promotes liver 

injury through inflammasome activation, or if sterile inflammation 

resolves injury through clearance of necrotic debris and 

stimulation of regeneration. Currently, there is limited information 

on the time course of cytokine formation or complement 

activation in human patients, especially in surviving (S) and 

non-surviving (NS) populations. This study addressed this deficit, 

with the hypothesis that NS patients would accumulate 

a greater degree of plasma cytokine levels, and have more 

severe complement depletion after APAP overdose. Circulating 

plasma levels of IL-1ß, TNF-α, IL-6, IL-8, IL-10, MCP-1, G-CSF, 

and GRO were measured by multi-plex ELISA and complement 

C3 was measured by ELISA in healthy volunteers (n=8), 

patients with APAP overdose but no increase in liver enzymes 

(n=10), and S (n=13) and NS (n=8) APAP overdose patients 

with liver injury, for the first seven days after study admission. 

ALT levels were similar between S and NS groups (5,531 ± 

639 U/L v. 5,302 ± 1,139 U/L), although bilirubin was significantly 

greater in the NS group (4.1 ± 1.0 mg/dL v. 8.6 ± 

3.1 mg/dL). Plasma levels of pro-inflammatory cytokines such 

as IL-1ß, GRO, and TNF-α did not rise above control levels at 

any point. In contrast, both S and NS patients had persistent, 

significant elevations in G-CSF, IL-6, IL-8, and anti-inflammatory 

cytokines such as IL-10 and MCP-1. Moreover, levels of IL-6, 

IL-8, IL-10 and MCP-1 were significantly higher in NS patients 

compared to S patients at multiple time points. MCP-1 levels 

rose to the greatest degree in both S patients (6,134 ± 1,134 

pg/mL) and NS patients (7,074 ± 1,105 pg/mL); whereas 

IL-1ß accumulated the least in both S patients (81.4 ± 64.2 

pg/mL) and NS patients (39.2 ± 32.6 pg/mL). Plasma C3 

levels were depleted at the same rate in S and NS patients, but 

recovered only in S patients. In conclusion, these data support 

the hypothesis that NS patients become refractory to anti-inflammatory 

cytokine signals, which likely fail to resolve injury 

or stimulate regeneration. Furthermore, there is limited inflammasome 

activation in human patients, as IL-1ß levels never rise 

above baseline. Importantly, however, other cytokines may 

have predictive value. 

Disclosures: 

The following authors have nothing to disclose: Benjamin Woolbright, Mitchell R. 

McGill, Matthew R. Sharpe, Hartmut Jaeschke 

586 

Protective effects of connexin43 signaling in acetaminophen-induced 

liver injury 

Michaël Maes 1 , Mitchell R. McGill 2 , Tereza Cristina da Silva 3 , 

Margitta Lebofsky 2 , Isabel V. Pereira 3 , Joost Willebrords 1 , Sara 

Crespo Yanguas 1 , Anwar Farhood 4 , Maria Lucia Zaidan Dagli 3 , 

Hartmut Jaeschke 2 , Bruno Cogliati 3 , Mathieu Vinken 1 ; 1 In Vitro 

Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, 

Brussels, Belgium; 2 Department of Pharmacology, Toxicology and 

Therapeutics, University of Kansas Medical Center, Kansas City, 

KS; 3 Department of Pathology, School of Veterinary Medicine 

and Animal Science, University of São Paulo, São Paulo, Brazil; 

4 Department of Pathology, St. David’s North Austin Medical Center, 

Austin, MO 

Background and aims: Being goalkeepers of liver homeostasis, 

gap junctions are also involved in hepatotoxicity. However, 

their role in this process is ambiguous, as gap junctions can act 

as both targets and effectors of liver toxicity. This particularly 

holds true for drug-induced liver insults. In the present study, the 

role and functional relevance of connexin32 and connexin43, 

the building stones of liver gap junctions, were investigated 

in acetaminophen-induced hepatotoxicity. Methods: C57BL/6 

mice were overdosed with acetaminophen followed by analysis 

of the expression and localization of connexins as well as 

monitoring of hepatic gap junction functionality. Furthermore, 

acetaminophen-induced liver injury was compared between 

mice genetically deficient in either connexin32 or connexin43 

and wild type animals. Evaluation of the toxicological response 

was based on a set of clinically relevant parameters, including 

protein adduct formation, histopathological examination, measurement 

of alanine aminotransferase, cytokines, reduced and 

oxidized glutathione, and analysis of proliferating cell nuclear 

antigen expression. Results: It was found that gap junction communication 

deteriorates upon acetaminophen intoxication in 

wild type mice, which was associated with a switch in mRNA 

and protein production from connexin32 to connexin43. Furthermore, 

connexin43-deficient animals showed increased 

liver cell death, inflammation and oxidative stress in comparison 

with wild type counterparts, whereas the opposite seems 

to hold true for mice lacking connexin32. Conclusion: These 

results suggest that hepatic connexin43-based signaling protects 

against acetaminophen-induced liver toxicity. 

Disclosures: 

The following authors have nothing to disclose: Michaël Maes, Mitchell R. 

McGill, Tereza Cristina da Silva, Margitta Lebofsky, Isabel V. Pereira, Joost 

Willebrords, Sara Crespo Yanguas, Anwar Farhood, Maria Lucia Zaidan Dagli, 

Hartmut Jaeschke, Bruno Cogliati, Mathieu Vinken 

587 

The Hepatic “Matrisome” Responds Dynamically to 

Inflammatory Injury: Proteomic Characterization of the 

Transitional ECM Changes in the Liver 

Christine E. Dolin 1 , Veronica L. Massey 1 , Lauren G. Poole 1 , 

Deanna L. Siow 1 , Michael L. Merchant 2 , Daniel W. Wilkey 2 , 

Gavin E. Arteel 1 ; 1 Pharmacology and Toxicology, University of 

Louisville, Louisville, KY; 2 Medicine, University of Louisville, Louisville, 

KY 

Background. The impact of the changes to the hepatic extracellular 

matrix (ECM) in the context of collagen accumulation 

during fibrosis are well known. However, the hepatic ECM 

consists of a myriad of biologically relevant proteins beyond 

collagen; furthermore, changes to the hepatic ECM are not 

restricted to fibrosis. Indeed, transitional changes to the hepatic 

ECM during inflammatory liver injury may be critical in the 

balance between restitution versus accumulation of damage.


The goal of this work was to develop a new proteomic method 

to characterize the hepatic ECM (“matrisome”) and document 

the impact of ethanol (EtOH) and lipopolysaccharide (LPS) on 

this compartment. Methods. Liver sections were solubilized 

in a series of increasingly rigorous extraction buffers (NaCl, 

SDS, GnHCl), to separate proteins by ‘age’ (e.g., crosslinking). 

Protein fragments in the extracts were identified by LC-MS/ 

MS. ECM proteins were identified, categorized by primary 

role and compared qualitatively and quantitatively between 

groups. This method was validated using liver sections from 

mice administered CCl 4 

for 4 weeks to establish fibrosis. The 

effects of 6 weeks EtOH diet and/or 24 hours LPS on the patterns 

of the matrisome were subsequently determined. Results. 

The proteomic approach validated an increase in collagen I 

protein in CCl 4 

–exposed livers; moreover, several other ECM 

proteins and collagens (e.g., III, IV and V) were identified to be 

changed by CCl 4 

exposure. Additionally, this approach indicated 

that the matrisome changed dramatically after steatosis 

(EtOH) and/or inflammation (LPS). EtOH caused a dramatic 

~30% increase in the number of matrisome proteins with the 

least change in the pellet fraction and collagen category; LPS 

produced a similar response. The enhancement of LPS-induced 

liver damage by EtOH preexposure demonstrated unique protein 

changes. The distribution of proteins across the extracts 

also changed in some proteins changed in response to insult, 

suggesting changes in their ‘age.’ Many proteins that did not 

qualitatively change with ethanol, LPS, and/or the combination 

quantitatively changed. Conclusions. These results suggest 

that this approach can document qualitative and quantitative 

changes to the hepatic ECM proteome. This proteomic technique 

reveals the intricacies of transitional ECM remodeling, 

including new players that may contribute to the mechanisms 

by which ethanol enhances hepatic inflammatory injury. These 

protein players likely include novel candidates for biomarkers 

and drug targets and will be examined more closely in future 

studies. Supported by NIH/NCI (5R25CA134283-03) and 

NIH/NIAAA (1R01AA021978-01). 

Disclosures: 

The following authors have nothing to disclose: Christine E. Dolin, Veronica L. 

Massey, Lauren G. Poole, Deanna L. Siow, Michael L. Merchant, Daniel W. 

Wilkey, Gavin E. Arteel 

588 

Golgi Stress Response Is Associated with Anti-HIV 

drug-induced ER Stress and Liver Cell Injury 

Hui Han, Cheng Ji; Medicine, Univ Southern California, Los Angeles, 

CA 

Background & Aim Certain antivirals commonly used in the 

Highly Active Antiretroviral Therapies for HIV-infected patients 

cause liver injury. Previously we found that endoplasmic reticulum 

(ER) stress contributes to the drug-induced liver injury. Here 

we further investigated whether Golgi stress response underlies 

the drug-induced hepatotoxicity. Methods HepG2 cells were 

treated with two HIV protease inhibitors, ritonavir (RIT) and 

lopinavir (LOP), ER stress-inducing agents, tunicamycin (Tm) 

or thapsigargin (Tg), or Golgi stress-inducing agents, sodium 

monesin (SM) or Benzyl 2-acetamido-2-deoxy-α-D-galactopyranoside 

(BADG). Cell death rate was assessed with Hochest 

blue/Syntox green double staining or Annexin V staining. 

Golgi stress response and ER stress response were analyzed 

with respective molecular markers. Results: RIT and LOP-induced 

ER stress response in the HepG2 cells was comparable 

to that by Tm or Tg, which was indicated by the presence of 

XBP-1 alternative splicing, increased phosphorylation of IREa, 

and increased expression of GRP94, ATF6, and EDEM1. However, 

RIT and LOP induced cell death was not comparable to 

that by Tg or Tm. RIT and LOP induced 40% cell death whereas 

Tg or Tm induced 15-20% cell death. Correspondingly, ER 

stress-related death markers, CHOP and phosphorylated JNK, 

were significantly higher in the RIT and LOP treated cells than in 

Tg or Tm treated cells. Further, RIT and LOP treatment increased 

three Golgi stress response markers, GCP60, HSP47 and TFE3, 

which was comparable to the Golgi stress response induced by 

SM or BADG. The Golgi stress response was not detected in 

Tg or Tm treated cells. The SM-induced Golgi stress led to 20% 

cell death based on the Annexin V analysis. Conclusion Our 

results suggest for the first time that both ER stress and Golgi 

stress contribute to the antiviral RIT and LOP induced liver cell 

death injury (supported by R01AA018612 NCE). 

Disclosures: 

The following authors have nothing to disclose: Hui Han, Cheng Ji 

589 

Acetaldehyde Triggers HCV-Induced Hepatitis Progression: 

A Proposed Mechanism 

Murali Ganesan 1,2 , Larisa Y. Poluektova 4,2 , Jinjin Zhang 3 , Tatiana 

Bronich 3 , Terrence M. Donohue 1,2 , Kusum K. Kharbanda 1,2 , Dean 

J. Tuma 1,2 , Natalia A. Osna 1,2 ; 1 Internal Medicine, University 

of Nebraska Medical Center, Omaha, NE; 2 Research, Veterans 

Affairs Nebraska-Western Iowa Health Care System, Omaha, NE; 

3 College of Pharmacy, University of Nebraska Medical Center, 

Omaha, NE; 4 Pharmacology and Experimental Neuroscience, University 

of Nebraska Medical Center, Omaha, NE 

Background: In HCV-infected patients, alcohol consumption 

accelerates hepatitis progression. For many years, the mechanism 

of this event was attributed to an ability of ethanol to 

enhance HCV RNA replication. This conclusion was mainly 

based on in vitro experimental data obtained from ethanol 

non-metabolizing hepatoma cells and was not supported by 

consistent results from HCV patient trials. Here, we investigated 

the pathogenic role of ethanol metabolism-mediated regulation 

of HCV RNA levels in the acceleration of liver injury. Methods 

and Results: HCV RNA was quantified by RT-PCR in HCV-infected 

CYP2E1 + Huh7.5 (RLW) cells exposed for 1-48 hrs to an 

external acetaldehyde (Ach) generating system (AGS). Unexpectedly, 

after transient up-regulation of HCV RNA levels after 

24 hrs of AGS exposure, we observed a decline in HCV RNA 

after 48 hrs of treatment with Ach. This finding was confirmed 

by the reduction of HCV core protein expression in these cells. 

The Ach-induced decrease of intracellular viral content was not 

related to leakage of viral particles out of cells, as since HCV 

infectivity in supernatants was blocked. In attempting to explain 

these events, we postulate that HCV infection pre-sensitizes cells 

to Ach-induced apoptosis, which depletes HCV + hepatocytes. 

Indeed, the cleavage of caspase 3 and of PARP were more 

robust in AGS-exposed HCV + than in HCV - cells after 48 hr. 

Importantly, suppression of Ach-triggered apoptosis in infected 

cells by the pan-caspase inhibitor prevented the decrease of 

HCV RNA by Ach, indicating that depletion of HCV + cells by 

Ach was caspase-dependent. To assess the pathogenic role of 

HCV-containing apoptotic bodies (AB) in liver inflammation, 

we generated AB from HCV + and HCV - RLW cells and incubated 

them with human primary macrophages. There were no 

differences in mRNA levels of TNFα, TGFβ and IL-12 when AB 

from both sources were used; however, IL-18, IL-1β and IL-10 

mRNAs were induced at higher levels by HCV + AB. Shaping 

of cytokine production in macrophages by AB from HCV + 

cells was virus-mediated, as the presence of HCV proteins in 

these AB was confirmed by Western blot. In conclusion, HCV 

sensitizes liver cells to pro-apoptotic effects of Ach, ultimately


generating higher levels of HCV-expressing AB. Engulfment of 

HCV + AB by liver macrophages increases the production of 

pro-inflammatory cytokines, thereby exacerbating liver injury 

in HCV + alcohol abusers. 

Disclosures: 

The following authors have nothing to disclose: Murali Ganesan, Larisa Y. Poluektova, 

Jinjin Zhang, Tatiana Bronich, Terrence M. Donohue, Kusum K. Kharbanda, 

Dean J. Tuma, Natalia A. Osna 

590 

Mitochondrial Fission Factor (MFF) is induced and binds 

to Sab (SH3BP5) in acetaminophen (APAP) and other 

JNK dependent hepatotoxicities 

Sanda Win, Tin A. Than, Neil Kaplowitz; Medicine, GI/Liver, USC 

KSOM, LosAngeles, CA 

SAB is a mitochondrial membrane binding target of JNK which 

mediates impaired mitochondrial function, increased ROS, 

and sustained JNK activation in hepatotoxicity from acetaminophen 

(APAP), TNF, palmitic acid, and ER stressors. We have 

previously shown that mitochondrial translocation of DRP-1, 

the mediator of mitochondrial fission and a key factor in cell 

death, is downstream of the interaction of P-JNK and Sab. 

MFF is the binding target of DRP-1 in the outer membrane of 

mitochondria. Therefore, our aim was to elucidate the role 

of Sab in mitochondrial fission. We immunoprecipitated (IP) 

Sab from mitochondrial extracts of livers of control and APAP 

treated mice (2 hours). MFF was identified in the IP and greatly 

increased after APAP, both by western blot and proteomic 

analysis. Under basal conditions, low levels of MFF are found 

in mitochondria but levels rapidly increased within one hour 

after APAP. Proteinase K treatment of intact mitochondria confirmed 

that MFF was exclusively in the outer membrane. The 

increase in MFF was not inhibited by actinomycin D suggesting 

that MFF is stabilized by P-JNK and Sab rather than by induction 

of transcription. MFF was also induced in other JNK/Sab 

dependent toxicity models such as tunicamycin, palmitic acid 

and TNF/galactosamine. Sab knockdown or liver conditional 

knockout of Sab abrogated the increase of MFF and prevented 

DRP-1 translocation to mitochondria. However, basal levels of 

MFF were not altered in the absence of Sab. We previously 

showed that knockdown of DOK4, an inner membrane Src 

binder, prevented mitochondrial inactivation of Src, mitochondrial 

dysfunction, increased ROS, sustained JNK activation, 

and cell death from APAP and TNF/galactosamine. Knockdown 

of DOK4 had no effect on Sab levels but prevented 

the induction of MFF suggesting that P-JNK/Sab mediated 

mitochondrial dysfunction leads to increased to MFF levels. In 

conclusion, the interaction of P-JNK and Sab leads to induction 

of MFF, enhancing mitochondrial fission. MFF appears to be 

stabilized by mitochondrial dysfunction and binding to Sab. 

Disclosures: 

The following authors have nothing to disclose: Sanda Win, Tin A. Than, Neil 

Kaplowitz 

591 

Hormesis in cholestatic liver disease; preconditioning 

with low bile acid concentrations protects against bile 

acid-toxicity 

Esther M. Verhaag, Manon Buist-Homan, Han Moshage, Klaas 

Nico Faber; Gastroenterology and Hepatology, University of 

Groningen, Groningen, Netherlands 

INTRODUCTION: Cholestasis is characterized by accumulation 

of bile acids and inflammation, causing hepatocellular damage, 

eventually resulting in liver failure. Bile acids induce apoptotic 

and necrotic cell death, however, hepatocytes are able to 

adapt towards a hostile environment. This suggests a hormetic 

response toward bile acids limits hepatocyte cell death during 

cholestasis. AIM: To investigate in vitro the mechanisms that 

underlie the hormetic response that protect hepatocytes against 

experimental cholestatic conditions. METHODS: HepG2.rNTCP 

cells were preconditioned (24 h) with sub-apoptotic concentrations 

(0.1-50 mM) of various bile acids, the superoxide donor 

menadione, the pro-inflammatory cytokine TNF-α or the FXR 

agonist GW4064, followed by a challenge with the apoptosis-inducing 

bile acid glycochenodeoxycholic acid (GCDCA; 

200 mM for 4 h). Levels of caspase-3/7 activity (apoptosis), 

cellular LDH leakage (necrosis) and the bile acid exporter BSEP 

(ABCB11) mRNA were analyzed. RESULTS: Preconditioning 

with the pro-apoptotic bile acids GCDCA or taurocholic acid 

(TCA), and the protective bile acids (tauro)ursodeoxycholic 

acid (TUDCA, UDCA) reduced GCDCA-induced caspase-3/7 

activity in HepG2.rNtcp cells. None of the bile acid-preconditioning 

conditions induced cellular LDH leakage from GCD- 

CA-challenged HepG2.rNtcp cells above control (untreated 

cells) levels. In contrast, preconditioning with sub-apoptotic 

concentrations of cholic acid (CA) potentiated GCDCA-induced 

apoptosis and similar trends were observed for menadione 

and TNF-α preconditioning. CDCA preconditioning did not 

change GCDCA-induced levels of caspase-3/7. The hormetic 

effect of GCDCA preconditioning was concentration- and 

time-dependent, providing significant protection against the 

apoptotic GCDCA challenge after at least 12 h preconditioning 

with concentrations of GCDCA as low as 1 mM. GCDCA 

and CDCA preconditioning both enhanced mRNA levels of 

ABCB11. The FXR agonist GW4064 more potently induced 

ABCB11 mRNA levels, but did not cause a significant reduction 

in GCDCA-induced caspase-3/7 activity, though a trend 

towards a partial suppression was observed. CONCLUSIONS: 

Sub-toxic concentrations of bile acids in the range that occur 

under normal physiological conditions protect HepG2.rNtcp 

cells against GCDCA-induced apoptosis, which is largely independent 

of FXR signaling. Thus, low concentrations bile acids 

enable hepatocytes to withstand sudden increases in bile acid 

concentrations and this may explain why only limited levels of 

apoptotic cell death is observed in obstructive cholestasis. 

Disclosures: 

The following authors have nothing to disclose: Esther M. Verhaag, Manon 

Buist-Homan, Han Moshage, Klaas Nico Faber 

592 

A protective role of C/EBP homologous protein in acetaminophen-induced 

acute hepatocyte damage in mice 

Tsutomu Matsubara 1 , Yuga Teranishi 2 , Kazuki Nakatani 1 , Norifumi 

Kawada 2 , Kazuo Ikeda 1 ; 1 Department of Anatomy and 

Regenerative Biology, Graduate School of Medicine, Osaka City 

University, Osaka, Japan; 2 Department of Hepatology, Graduate 

School of Medicine, Osaka City University, Osaka, Japan 

Background: Acetaminophen (APAP) overdose causes acute 

liver failure as a result of increased oxidative stress followed by 

hepatocellular necrosis. C/EBP homologous protein (CHOP) 

expression is induced the liver after exposure to APAP, but 

the role of CHOP in the APAP-induced liver injury is not fully 

understood. This study investigated whether CHOP acts on 

the APAP-induced liver injury using wild-type and Chop-null 

mice. Methods: Acute liver injury was induced in wild-type 

and Chop-null mice by administrating acetaminophen (APAP, 

300 mg/kg). The mice were sacrificed at six hours after the 

APAP injection. Liver damage was evaluated by measuring 

serum ALT levels and liver histology. Hepatic mRNA and pro-


tein levels were determined by quantitative PCR and western 

blot analysis. Results: Six hours after 300 mg/kg of APAP 

injection, serum ALT levels were 3831 and 7897 U/L and 

relative hepatic necrosis area was 22.7 and 47.1% of view 

field in wild-type and Chop-null mice, respectively. The Chopnull 

mice showed a severer liver injury than the wild-type mice. 

When phosphorylations of AKT, ERK, JNK, and p38 proteins 

in the livers were investigated, APAP-enhanced JNK phosphorylation 

was reduced in the Chop-null mice, compared to the 

wild-type. In addition, MKK4 and JUN phosphorylations were 

also reduced in Chop-null mice, suggesting that JNK signal 

was attenuated in the Chop-null livers. Increased mRNA levels 

of TIFA, an NF-kB activator, and enhanced phosphorylation of 

FOS, an NF-kB-target protein, were observed in the livers of 

the Chop-null mice, suggesting that NF-kB signal was accelerated 

in the Chop-null livers. Hepatic ratio of LC3-II protein 

to LC3-I protein, an indicator of autophagy, was increased in 

the wild-type mice after the APAP injection (2.4 fold), but the 

increase was diminished in the Chop-null mice. Furthermore 

protein levels of hepatic p62 (also called SQSTM1), protein 

decomposed under autophagy, were higher in the Chop-null 

mice than that in the wild-type mice. There results strongly indicated 

a view that autophagy signal was attenuated in the livers 

of Chop-null mice Conclusions: In this study, molecular linkage 

of CHOP-JNK-autophagy was identified as a protective factor 

in APAP-induced liver injury. Although further study is required, 

accelerated NF-kB signal in Chop-null livers may interfere with 

the protective role of the JNK-autophagy. 

Disclosures: 



The following authors have nothing to disclose: Tsutomu Matsubara, Yuga Teranishi, 

Kazuki Nakatani, Kazuo Ikeda 

593 

The Effect of Treatment with Chemical Chaperones and 

Zinc Acetate on Hepatocytes Treated with Excess Copper 

Shinji Oe, Koichiro Miyagawa, Yuichi Honma, Masaru Harada; 

School of Medicine, University of Occupational and Environmental 

Health, Japan., Fukuoka, Japan 

[Purpose] Copper is an essential trace element in human activity. 

But excess copper gives harm to human activity. Wilson 

disease is a genetic disorder characterized by excess copper 

deposition in various organs. Excess copper-derived oxidants 

contribute to progression of liver disease in Wilson disease. 

Oxidative stress induces accumulation of abnormal proteins. 

The endoplasmic reticulum (ER) is subcellular organelle which 

plays the role of proper protein folding. Accumulation of misfolded 

proteins disturbs ER homeostasis resulting in ER stress. 

Zinc acetate has been commonly used to treat Wilson disease, 

because it increases metallothionein production and inhibits 

absorption of copper in the intestinal epithelia. The effects of 

chemical chaperones and zinc acetate on copper-induced hepatotoxicity 

have not been examined. In this study, we evaluated 

the effects of chemical chaperone and zinc acetate on copper-induced 

disruption of cell homeostasis. [Methods] We used 

human hepatoma cell line (Huh7) and immortalized human 

hepatocyte cell line (OUMS29). The following materials were 

used: copper sulfate; acetyl-leucyl-leucyl-norleucinal and epoxomicin 

as proteasome inhibitors (PIs); bathocuproine disulfonate 

as copper chelator; n-acetyl-l-cysteine; zinc acetate; 4-phenylbutyrate 

(PBA) and ursodeoxycholic acid (UDCA) as chemical 

chaperones. We detected the reactive oxygen species using 

2’,7’-dichlorodihydrofluorescein . 

We analyzed copper-induced 

ER stress by immunoblotting for ER stress markers, including 

phospho α-subunit of eukaryotic initiation factor 2 and X-box 

binding protein 1. Hepatocyte apoptosis treated with copper 

with or without PI was determined by detection of cleaved poly 

ADP-ribose polymerase and cleaved caspase 3 by immunoblotting 

analysis and immunofluorescence staining, respectively. 

Furthermore, we examined cell proliferation on excess copper 

by immunofluorescence staining using Ki67. [Results] Excess 

copper induced oxidative stress and ER stress. Chemical chaperones 

and zinc acetate reduced copper-induced oxidative 

stress and ER stress. Co-treatment of copper and PI exacerbated 

copper-induced apoptosis. Although excess copper leaded to 

reduction in cell proliferation, chemical chaperones rescued 

the proliferation block in cells treated with copper. However 

zinc acetate did not exert this effect. [Conclusions] Excess copper 

induced hepatotoxicity. Chemical chaperones, PBA and 

UDCA, and zinc acetate reduced hepatotoxicity. These results 

suggest that chemical chaperones may have beneficial effects 

for the treatment of Wilson disease. On the other hand, zinc 

acetate used as the treatment of Wilson disease may inhibit 

liver injury directly. 

Disclosures: 

The following authors have nothing to disclose: Shinji Oe, Koichiro Miyagawa, 

Yuichi Honma, Masaru Harada 

594 

Sumoylation Regulates Lipopolysaccharide-induced 

Macrophage Activation Modulating Ubiquiting conjugating 

enzyme 9 Phosphorylation in Mouse Liver and 

Kupffer Cells 

Maria Lauda Tomasi 2,1 , Komal Ramani 2 , Minjung Ryoo 2 ; 1 Medicine, 

University of Southern California, Los Angeles, CA; 2 Medicine, 

Cedars-Sinai Medical Center, Los Angeles, CA 

Propose of study: Activation of macrophages by lipopolysaccharide 

(LPS), a conserved component of the Gram-negative 

bacterium’s outer membrane, leads to production of a large 

number of immunoregulatory molecules including tumor necrosis 

factor α (TNF-α), interleukin 1β (IL-1β), interleukin 6 (IL-6), 

arachidonic acid, as well as reactive oxygen species (ROS). 

LPS level is increased in the blood of alcohol abusers and ethanol-fed 

rodents. The small ubiquitin-related modifier (SUMO) 

family of proteins is a more recently discovered posttranslational 

modification. Cellular stress arising from ROS is a robust stimulus 

for protein SUMOylation. SUMOylation is widely deemed 

as a protective mechanism; loss of SUMO conjugation reduces 

cell and organism viability. Ubiquitin conjugating enzyme 9 

(UBC9) is the only SUMO E2 enzyme. We demonstrated that 

UBC9 is phosphorylated by Cyclin-dependent kinase 1 (CDK1) 

and is more stable when phosphorylated. Furthermore, we 

recently found that LPS lowers UBC9 and SUMO expression 

in RAW cells, nevertheless the mechanism is still unknown. The 

aim of this study was to examine the role of Ubc9-mediated 

sumoylation in LPS-activated macrophages and elucidate the 

molecular mechanism(s). Methods: Studies were done using in 

vivo LPS-treated mice, primary mouse Kupffer and RAW cells. 

Real-time PCR and Western blotting measured mRNA and protein 

levels, respectively. UBC9 phosphorylation was assessed 

by phos-tagÔ SDS-PAGE. Results: LPS treatment increased ROS 

production (H 2 

O 2 

) in RAW cells, while UBC9 knockdown alone 

or co-treatment with LPS completely prevented LPS-induced 

ROS production. UBC9 siRNA alone or with LPS co-treatment 

induced apoptosis and increased IL-1β, iNOS, IL-6 and TNF-α 

mRNA levels in RAW cells; however, when LPS and UBC9 

siRNA were combined, they further raised these inflammatory 

signals. We found that LPS treatment lowers UBC9 protein


level in the LPS-treated mouse liver, primary Kupffer and RAW 

cells as compared with controls, whereas UBC9 mRNA level 

decreased minimally. Phos-tagÔ SDS-PAGE showed that LPS 

increased UBC9 phosphorylation in mouse liver and primary 

Kupffer cells. These results support a potential key mechanism 

for LPS that induces pro-inflammatory cytokines through lowering 

UBC9 expression, raising its phosphorylation. Conclusions: 

The novel finding that LPS lowers UBC9 protein level has 

important implications since this facilitates the pro-inflammatory 

response. A better understanding of molecular mechanisms on 

how LPS treatment influences sumoylation-regulated cytokines 

in Kupffer cells, will open a crucial area of investigation providing 

new targets for therapy in alcoholic liver diseases. 

Disclosures: 

The following authors have nothing to disclose: Maria Lauda Tomasi, Komal 

Ramani, Minjung Ryoo 

595 

A comparative analysis of the Spanish and Latin-American 

prospective drug-induced liver injury (DILI) networks 

Fernando Bessone 1 , Nelia Hernández 2 , Adriana Sanchez Ciceron 

2 , Maria Di Pace 2 , Gisela Gualano 3 , Marco Arrese 4 , Alex Ruiz 4 , 

Aurora Loaeza del Castillo 5 , Marcos A. Girala 6 , Enrique Carrera 7 , 

Maribel Lizarzábal 8 , Edgardo Mengual 8 , Javier Brahm 9 , Juan P. 

Arancibia 9 , Federico C. Tanno 1 , Milagros B. Davalos-Moscol 10 , 

Raymundo Paraná 11 , M I. Schinoni 11 , Nahum Méndez-Sanchéz 12 , 

Miguel E. Garassini 13 , Rodrigo L. Zapata 14 , I. Medina-Cáliz 15 , 

Andrés González-Jiménez 15 , Mercedes Robles Diaz 15 , C. Stephens 

15 , A. Ortega 15 , Judith Sanabria 15 , Miren García-Cortés 15 , 

B. Garcia-Muñoz 15 , M. I. Lucena 15 , Raul J. Andrade 15 ; 1 Hospital 

Provincial del Centenario, Rosario, Argentina; 2 Hospital de 

Clínicas, Montevideo, Uruguay; 3 Hospital Posadas, Buenos Aires, 

Argentina; 4 Pontificia Universidad Católica de Chile, Santiago, 

Chile; 5 Instituto Nacional de Ciencias Médicas y Nutrición “Salvador 

Zubirán”, Hospital General de México, Ciudad de Mexico, 

Mexico; 6 Hospital de Clínicas, Asunción, Paraguay; 7 Hospital de 

Especialidades Eugenio Espejo, Quito, Ecuador; 8 Hospital Universitario 

de Maracaibo, Maracaibo, Venezuela, Bolivarian Republic 

of; 9 Hospital Clínico Universidad de Chile, Santiago, Chile; 

10 Hospital Rebagliati, Facultad de Medicina, Universidad San 

Martin de Porres, Lima, Peru; 11 Hospital Universitario Prof. Edgard 

Santos, Universidad Federal da Bahía, Salvador de Bahía, Brazil; 

12 Fundación Clínica Médica Sur, Ciudad de Mexico, Mexico; 

13 Hospital Universitario de Caracas, Caracas, Venezuela, Bolivarian 

Republic of; 14 Universidad de Chile, Clínica Alemana de 

Santiago, Santiago, Chile; 15 Instituto de Investigación Biomédica 

de Málaga (IBIMA), Hospital Universitario Virgen de la Victoria, 

Universidad de Málaga, CIBERehd, Málaga, Spain 

Background: DILI characteristics concerning phenotype and 

involved drugs or other toxic compounds can vary between 

individuals and possibly between different geographic populations. 

We aimed to compare all DILI cases included in the 

ongoing Spanish and Latin-American DILI Network that share 

the same inclusion criteria and operational procedures. Material 

and methods: Demographics, clinical parameters and 

causative agents were compared between 200 Latin-American 

and 867 Spanish DILI cases. Results: The mean age of DILI 

development differed between the two registries with 51 years 

in LatinAmerica and 54 years in Spain (p=0.02). Females predominated 

among the LatinAmerican cases (59%) compared 

to the Spanish cases (49%) (p=0.01). Duration of treatment 

and time to onset were higher in LatinAmerican cases (127 vs 

88 days, p


Drug properties (Liver Toxicity Knowledge Base, Drug Discovery 

Today 2011) and host factors were compared in DO vs. 

NDO. Results: Among the 680 cases, 159 cases (22%) manifested 

DILI with 2 to 82 days’ delay after the treatment cessation 

(i.e., DO) while 521 (77%) manifested during the drug treatment 

(i.e., NDO). 57% of DO cases and 13% of NDO were 

Amoxicillin-clavulanate (AMOX/CA) cases. After excluding the 

AMOX/CA cases, DO cases had shorter treatment duration 

(median: 11 vs. 45 days, p=50% were less prevalent in 

DO cases (67% vs. 84%, p=0.003) also drugs with un-metabolized 

drug excretion >=50% were more prevalent in DO cases 

(25% vs. 9%, p=0.0011). Regarding host manifestations, 

eosinophilia was more prevalent in DO cases (30% vs. 19%, 

p=0.036), while positive autoantibody was more prevalent in 

NDO cases (25% vs. 11%, p=0.024). NDO cases were more 

associated with chronic underlying diseases (82% vs. 60%, 

p 50% loss, n=14]. Results: The 28 subjects 

with bile duct loss included 14 men [5 severe] and 14 women 

[9 severe]. The median age was 52.5 [range 11-87] years 

those with severe duct loss were younger [47.4 vs 59.7 years, 

p=0.04]. Cases were attributed to 20 different agents: 3 to 

amoxicillin/clavulanic acid; 3 to temozolomide; 3 to botanicals 

[Cactus nopal; Artemisia annua; unknown]; 2 to azithromycin; 

and 1 each due to cefalexin, cefazolin, levofloxacin, moxifloxacin, 

allopurinol, enalapril, hydralazine, infliximab, lenalidomide, 

thalidomide, montelukast, olanzapine, quetiapine, 

lansoprazole, omeprazole, metoclopramide and mesalazine. 

Median latency to onset was 35 [IQR 19-91] days. The typical 

clinical presentation was immuno-allergic, rather than autoimmune-like 

disease and was usually cholestatic or mixed, with 

median R-value= 2.1 [range 0.1-8.0]. In addition to bile duct 

paucity, hepatic pathology showed reactive bile duct injury 

and mild portal lympho-histiocytic inflammatory infiltrates. 

Eosinophils were noted in 9 biopsies, but peripheral eosinophilia 

at disease onset was noted in only 6 patients. 7 of 14 

subjects with mild and 8 of 14 with severe injury transitioned 

to chronic ongoing injury [laboratory, imaging or clinical evidence 

of persisting liver damage > 6 months after onset]. The 

likelihood of dying or requiring liver transplantation within 6 

months of onset was greater in those with severe bile duct 

loss (4/14, 29%) than in mild duct loss (0/14, 0%, p=0.05] 

and also greater than in those without duct loss on biopsy 

(22/246, 8.9%, p=0.04). Conclusions: Paucity of bile ducts 

is an uncommon (6%) histologic feature of acute drug-induced 

liver injury, can be caused by diverse agents, both drugs and 

botanicals, and often leads to evidence of chronic liver injury, 

which, when severe, may lead to premature death or liver 

transplantation. Pathogenesis remains imperfectly understood, 

but host immune reactions, perhaps influenced by genetic or 

environmental factors, appear likely; however, the key antigen[s] 

remain unknown. 

Disclosures: 

Herbert L. Bonkovsky - Advisory Committees or Review Panels: Recordati Rare 

Chemicals, Clinuvel, Inc.; Consulting: Alnylam, Inc, Clinuvel, Inc., Clinuvel, Inc.; 

Grant/Research Support: Gilead Sciences 

Mark W. Russo - Grant/Research Support: Merck, Salix; Speaking and Teaching: 

janssen, Gilead, ABBVIE, Salix 

The following authors have nothing to disclose: David E. Kleiner, Jiezhun Gu, 

Joseph A. Odin, Victor J. Navarro, Maricruz Vega, Jay H. Hoofnagle 

598 

Caspase Inhibition Switched TNF-α-Induced Apoptosis 

to Necroptosis But Not Autophagic Cell Death In 

Hepatocytes and Mouse Livers 

Hong-Min Ni, Xiaojuan Chao, Mitchell R. McGill, Hartmut 

Jaeschke, Wen-Xing Ding; Pharmacology, Toxicology and Therapeutics, 

The University of Kansas Medical Center, Kansas City, KS 

Hepatocyte death (apoptosis and necrosis) and survival (such 

as autophagy) are integrated and play important roles in many 

liver diseases such as endotoxin and alcohol-induced liver 

injury. How these different cell death and survival pathways 

regulate each other remains elusive. We previously demonstrated 

that when the cell survival NF-κb pathway was blocked 

by a general transcription inhibitor actinomycin D (ActD), tumor 

necrosis factor-a (TNF-a) induced apoptosis in primary cultured 

mouse hepatocytes, which was completely suppressed by a 

general caspase inhibitor, ZVAD. Surprisingly, in the present 

study, we found that TNF-a/ActD-treated hepatocytes died 

by a necrotic-like cell death evidenced by abundant cellular


vacuoles, propidium iodide positive stained intact nuclei and 

release of nuclear HMGB1 after a pro-longed treatment for up 

to 48 hours. TNF-a/ActD induced cleavage of receptor interacting 

protein kinase 1 (RIP1) and RIP3 as well as Beclin 1, key 

molecules that regulate necroptosis and autophagy, respectively. 

These cleavages were mediated by caspases because 

they were blocked by ZVAD. Necrostatin 1 (a RIP1 inhibitor) 

or hepatocytes isolated from RIP3 knockout mice partially 

protected against TNF-a/ActD+ZVAD-induced cell death, but 

antioxidants (N-acetylcysteine or MnSOD) and a JNK inhibitor 

(SP600125) had no effect. Using adeno-RFP-GFP-LC3 infected 

hepatocytes, we found that TNF-a/ActD+ZVAD decreased 

autophagic flux. Moreover, TNF-a/ActD+ZVAD-induced cell 

death in hepatocyte isolated from liver-specific Atg5 knockout 

mice was similar to wild type hepatocytes. These data suggest 

that TNF-a/ActD+ZVAD induced RIP1/3-mediated necroptosis 

but not autophagic cell death in vitro. Similar to primary 

hepatocytes, we found that ZVAD also protected against LPS/ 

galactosamine (GalN)-induced apoptosis and liver injury at 6 

hours in mice. However, this protection was lost at 24 hours 

and these mice developed severe hepatic necrosis, neutrophil 

infiltration and liver injury. Intriguingly, while pharmacological 

inhibition of RIP1 or RIP3 knockout mice suppressed LPS/Gal- 

N+ZVAD-induced necrosis, they did not protect against LPS/ 

GalN+ZVAD-induced liver injury due to increased caspase-independent 

apoptosis in these mouse livers. In conclusion, our 

results suggest that different cell death and cell survival pathways 

are closely integrated during TNF-a-induced liver injury 

when both caspase and NF-κb are blocked. Blocking caspase 

switched TNF-a-induced apoptosis to RIP1/3-dependent necroptosis 

but not autophagic cell death. Thus, results from our 

study also raise concerns on the safety of current ongoing clinical 

trials using caspase inhibitors. 

Disclosures: 

The following authors have nothing to disclose: Hong-Min Ni, Xiaojuan Chao, 

Mitchell R. McGill, Hartmut Jaeschke, Wen-Xing Ding 

599 

Mitochondrial-shaping proteins as specific biomarkers 

to distinguish alcohol from fat-induced liver toxicity 

Elena Palma 1 , Antonio Riva 1 , Satvinder Mudan 2 , Nikolai Manyakin 

2 , Roger Williams 1 , Shilpa Chokshi 1 ; 1 Institute of Hepatology- 

Foundation for Liver Research, London, United Kingdom; 2 The 

London Clinic, London, United Kingdom 

Alcoholic Liver Disease (ALD) and Non-Alcoholic Liver Disease 

(NAFLD) show similar pathological spectra and present common 

factors (lipotoxicity, oxidative stress), but differ in clinical 

aspects and outcomes. A clearer understanding of their 

pathogenesis would be helpful in a better management of these 

disorders. Morphological changes as abnormal enlargements 

in the hepatic mitochondria, particularly megamitochondria 

(MM) formation, have been reported in liver biopsies from ALD 

patients since the 1970s but are not associated with NAFLD. 

Mitochondrial architecture is intimately involved with their 

function and adapts rapidly in response to the needs of the 

cell with cycles of fusion (organelle binding) and fission (fragmentation). 

These changes are strictly regulated by the activity 

of the mitochondrial-shaping proteins (MSP). Our aim is to 

elucidate the role of mitochondrial dynamics, shape and MSP 

in ALD and NAFLD in order to identify early disease specific 

markers. Human precision cut liver slices and VL-17A cells 

(positive for ADH/CYP2E1) were cultured with ethanol (EtOH) 

and/or oleic/linoleic acid (FFA), to induce hepatotoxicity. 

Post-treatment cell viability, intracellular fat accumulation and 

mitochondrial functionality were evaluated and changes in the 

mitochondrial shape were assessed by confocal and electron 

microscopy; MSP expression and activation were analysed by 

Western blot. The toxic effect of EtOH and FFA was confirmed 

by an increase in the percentage of apoptosis or a reduction 

in ATP levels and an impairment of mitochondrial respiration. 

The two insults induced a very different mitochondrial phenotype, 

as well as differential intracellular fat accumulation. Fat 

was associated with a more pronounced toxicity and excessive 

mitochondrial fission, while EtOH alone showed a moderate 

effect. With EtOH exposure, the surviving cells showed a dramatically 

increased proportion of MM, and this was associated 

with a reduced activation of the master fission regulator 

Drp-1 (Dynamin related protein-1). In parallel, changes in MSP 

involved in mitochondrial fusion were observed with fat-induced 

hepatotoxicity. In conclusion, we demonstrate that hepatotoxic 

agents such as EtOH or fat affect the mitochondrial dynamics 

and shape differently, altering the fusion/fission equilibrium 

and MSP expression. In particular, we show MM formation in 

response to the chronic EtOH insult in hepatocytes that survive, 

suggesting this as an adaptive mechanism to avoid death and 

raising the potentiality for the use of MSP as therapeutic targets 

for ALD and disease-specific biomarkers potentially discriminating 

between the early toxic events observed in ALD and 

NAFLD. 

Disclosures: 

The following authors have nothing to disclose: Elena Palma, Antonio Riva, Satvinder 

Mudan, Nikolai Manyakin, Roger Williams, Shilpa Chokshi 

600 

Patients with childhood-onset primary sclerosing cholangitis 

harbor rare, deleterious variants in genes involved 

with cholestatic syndromes and generalized risk of PSC 

Brian D. Juran, Aliya Gulamhusein, Saurabh Baheti, Konstantinos 

Lazaridis; Mayo Clinic, Rochester, MN 

Purpose: Genome-wide association studies (GWAS) of primary 

sclerosing cholangitis (PSC) have linked common genetic variants 

to PSC risk; yet, rare genetic variants also likely impact 

this disease. Next-generation sequencing provides access to 

these variants; however, poor functional predictability and the 

expectation of low-penetrance among alleles not manifesting 

in disease until adulthood, poses a serious obstacle to meaningful 

studies. Here, we focus on patients with childhood-onset 

PSC, as they more-likely harbor highly-penetrant variants, and 

concentrate on protein-alterations among candidate genes with 

prior likelihood for PSC impact, to maximize functional prediction. 

This effort will advance our scant knowledge regarding 

childhood PSC, and lay the groundwork towards application of 

sequencing-based approaches to the at-large PSC population. 

Methods: 10 patients with PSC diagnosis prior to age 12 were 

identified in the PSC Resource of Genetic, Environment and Synergy 

Studies (PROGRESS). Exome capture was by Agilent Sure 

Select XT Human Exome +UTR v5 kit and sequencing was on 

an Illumina HiSeq2000. Alignment, calling of single nucleotide 

variants (SNVs), and QC was performed using an established 

pipeline based on the Genome Analysis Toolkit Best Practices 

workflow; annotation was performed using BioR. Missense or 

nonsense SNVs were filtered based on quality and inclusion 

in manually-curated lists of (a) 28 known cholestasis genes or 

(b) 15 PSC GWAS-implicated genes. Final filtering kept only 

rare SNVs (population minor allele frequency < 1.5%) and 

those expected to be functional by at least 1 of 3 predictive 

tools (SIFT, PolyPhen-2 or Mutation Taster). Results: 14 SNVs 

remained after filtering; 13 were single-instance alleles found 

in 1 patient and 1 was found in 2 patients (rs12371484 in 

SH2B3). 9 of the SNVs were from the list of known cholestasis


genes and located in the following genes ABCB4, ABCC2, 

ATP8B1, HSD3B7, NOTCH2 (1 SNV each) and SERPINA1, 

TJP2 (2 SNVs each). The 5 SNVs from the PSC GWAS list were 

located in the genes CD226, GPR35, MST1 (1 SNV each) and 

SH2B3 (2 SNVs). Overall, 9/10 childhood-onset PSC patients 

had at least one of the SNVs (4 patients with 1 SNV, 4 with 

2 SNVs and 1 patient with 3 SNVs). Conclusions: Patients 

with childhood-onset PSC harbor rare, deleterious variants in 

genes involved with cholestasis and risk of PSC, suggesting that 

disease in these patients may lie along a spectrum between 

syndromic forms of cholestatic disease and classical PSC. Additional 

studies including patients with more-typical disease onset 

will help to confirm this finding and may establish a role for 

these genes in adult disease. 

Disclosures: 

The following authors have nothing to disclose: Brian D. Juran, Aliya Gulamhusein, 

Saurabh Baheti, Konstantinos Lazaridis 

well that patients should be treated early in disease, when 

ALP and bilirubin levels first exceed ULN, and that patients be 

treated to the lowest levels possible to avoid liver transplant 

and premature death. 

Disclosures: 

David Jones - Consulting: Intercept, Pfizer, Novartis; Speaking and Teaching: 

Falk, Shire 


The following authors have nothing to disclose: Bettina E. Hansen, Willem J. 

Lammers, George F. Mells, Marco Carbone 

601 

Convergence of Two Predictive Models of Risk Reduction 

in Patients with Primary Biliary Cirrhosis (PBC) 

Bettina E. Hansen 1 , Willem J. Lammers 1 , David Jones 2 , Henk R. van 

Buuren 1 , George F. Mells 3 , Marco Carbone 4 ; 1 Erasmus MC, University 

Medical Center Rotterdam, Rotterdam, Netherlands; 2 Institute 

of Cellular Medicine, Newcastle University Medical School, 

Newcastle Upon Tyne, United Kingdom; 3 Academic Department 

of Medical Genetics, University of Cambridge, Cambridge, United 

Kingdom; 4 Division of Gastroenterology and Hepatology, Department 

of Medicine, Addenbrooke’s Hospital, Cambridge, United 

Kingdom 

Background: Two recent analyses on prognostic algorithms 

in large PBC cohorts provides a unique opportunity to test the 

replicability of effect. Objective: The goal of this study was to 

apply common assumptions to algorithms developed by the 

Global PBC and UK PBC cohorts to predict liver transplant-free 

survival, and to determine if the models are comparable in 

terms of quantified risk. Methods: The Global PBC cohort 

consists of 4,565 PBC patients treated with ursodeoxycholic 

acid (UDCA), recruited from 15 centers in 8 countries. An 

algorithm predicting liver transplant and all-cause mortality 

was created using Cox proportional hazards regression on 

a derivation sample (n=2488), and validated in a matched 

sample (n=1631). Predictor variables included: age, and 

serum bilirubin, alkaline phosphatase, albumin and platelet 

count measured 1 year after UDCA initiation. The UK PBC 

cohort (n=4022) was recruited from 150 National Health Service 

(NHS) Trusts across the UK and includes both patients 

treated and not treated with UDCA. An algorithm predicting 

liver transplant and liver-related death was derived from 2414 

patients and validated in 1608 patients. The predictor algorithm 

includes serum bilirubin, alkaline phosphatase, albumin, 

platelet count and ALT. The absolute risk for liver transplant and 

survival was calculated using equivalent fixed inputs for albumin 

(40 g/L or 1.143 of LLN=35) and platelets (120 X 10 9 /L); 

age set at 65 and ALT at 84 U/L. Bilirubin varied as a function 

of ULN: 1X = 17 mmol/L; 2X = 34 mmol/L; 3X = 51 mmol/L; 

4X = 68 mmol/L and ALP by 10 U/L, with range as a function 

of ULN: 1X = 150 U/L to 4X = 600 U/L. Results are shown in 

the figure below. Conclusion: The calculations showed comparability 

at bilirubin > 2ULN and near comparability at bilirubin 

at ULN, providing further compelling evidence that both ALP 

and bilirubin are reliable predictors of PBC outcomes. The convergence 

of data from these models argues strongly for a reassessment 

of treatment goals. These results indicate that patients 

with advanced disease are at highest need of treatment, as 

602 

Differentially Expressed Plasma miRNAs as Novel Biomarkers 

for the Early Detection of Primary Sclerosing 

Cholangitis and Cholangiocarcinoma 

Francesca Bernuzzi 2 , Francesco Marabita 4 , Ana Lleo 2 , Ilaria Bianchi 

2 , Massimiliano Mirolo 5 , Marco Marzioni 6 , Gianfranco Alpini 7 , 

Domenico Alvaro 8 , Kirsten M. Boberg 9 , Massimo Locati 5 , Guido 

Torzilli 10 , Lorenza Rimassa 11 , Fabio Piscaglia 12 , Xiao-Song He 1 , 

Patrick S. Leung 1 , Christopher L. Bowlus 3 , Guo-Xiang Yang 1 , M. 

Eric Gershwin 1 , Pietro Invernizzi 2 ; 1 Internal Medicine, University 

of California, Davis, CA; 2 Liver Unit and Center for Autoimmune 

Liver Diseases, Humanitas Clinical and Research Center, Milan, 

Italy; 3 Division of Gastroenterology and Hepatology, University of 

California at Davis, Davis, CA; 4 Unit of Computational Medicine, 

Department of Medicine, Karolinska Institute, Stockholm, Sweden; 

5 Department of Medical Biotechnologies and Translational Medicine, 

University of Milan, Milan, Italy; 6 Department of Gastroenterology, 

Universita Politecnica delle Marche, Ancona, Italy; 

7 Department of Medicine, Division of Gastroenterology, Texas 

A&M University Health Science Center, Temple, TX; 8 Department 

of Clinical Medicine, Division of Gastroenterology, University of 

Rome, Rome, Italy; 9 Medical Department, Rikshospitalet, Oslo, 

Norway; 10 Liver Surgery Unit, Department of Surgery, Humanitas 

Clinical and Research Center, Milan, Italy; 11 Medical Oncology 

and Hematology Unit, Humanitas Clinical and Research Center, 

Milan, Italy; 12 Department of Medical and Surgical Sciences 

DIMEC, Alma Mater Studiorum, University of Bologna, Bologna, 

Italy 

Background: Cholangiocarcinoma (CCA) is the second most 

common primary hepatic malignancy and a frequently fatal 

complication of primary sclerosing cholangitis (PSC) with the 

highest incidence within the first 2.5 years following the diagnosis 

of PSC. The lack of sensitive and specific biomarkers 

poses a major challenge in the early diagnosis of CCA in the 

setting of PSC. Specific plasma microRNAs (miRNAs) have 

been reported in a number of inflammatory and neoplastic 

conditions. The aim of the study was an intensive discovery 

phase to identify specific plasma miRNAs as diagnostic biomarkers 

for PSC and CCA. Methods: Ninety human plasma 

samples (30 PSC, 30 CCA, and 30 healthy subjects) were 

surveyed for the levels of 667 miRNAs by qRT-PCR to identify 

disease-associated candidate miRNAs (discovery phase). 

miRNAs that demonstrated an absolute log 2 

(fold change) > 

1, mean Ct < 30, and p < 0.05 were selected for validation. 

Candidates were validated in an independent cohort of 

130 samples (40 PSC, 40 CCA, 10 primary biliary cirrhosis


(PBC) and 40 healthy subjects). To evaluate the discriminating 

effect of plasma miRNAs, ROC curve were established for 

each validated miRNAs, and AUC was calculated from the 

logistic regression. Only miRNAs with AUC>0.70 were considered 

useful as biomarkers. Results: In the discovery phase, 

we identified 21 miRNAs with differential expression in PSC, 

33 in CCA, and 26 in both, when compared with healthy controls, 

of which 24 miRNAs demonstrated significantly diverse 

expression between PSC and CCA. Data from validation phase 

demonstrated that miR-200c was differentially expressed in 

PSC versus healthy controls, whereas miR-483-5p, and miR- 

194 were differentially expressed in CCA compared with 

healthy controls. Importantly, we also found 3 miRNA that were 

specifically deregulated in CCA when compared to PSC: miR- 

222 was upregulated and miR-16 and miR-195 were down 

regulated in CCA when compared to PSC. The combined ROC 

analysis of identified miRNAs from each disease group significantly 

increases the AUC value and the combination of 

these markers further improved the specificity and accuracy of 

diagnosis. Conclusions: This study is unique and an important 

discovery program that emphasizes the potential identification 

of plasma miRNAs as biomarkers of PSC and CCA. A panel 

of differentially expressed plasma miRNAs is identified in PSC 

and CCA when compared with healthy controls. Furthermore, 

our data provides a basis for the use of miRNAs as biomarkers 

not only for the diagnosis of PSC and for the early detection of 

CCA, but also novel therapeutic targets. Future analyses will 

depend on larger cohorts of patients. 

Disclosures: 

Lorenza Rimassa - Board Membership: Eli Lilly, Merck Serono 

Fabio Piscaglia - Advisory Committees or Review Panels: Bayer ; Consulting: 

Bracco, Eisai; Grant/Research Support: Esaote 




Sciences, Inc 

The following authors have nothing to disclose: Francesca Bernuzzi, Francesco 

Marabita, Ana Lleo, Ilaria Bianchi, Massimiliano Mirolo, Marco Marzioni, 

Gianfranco Alpini, Domenico Alvaro, Kirsten M. Boberg, Massimo Locati, Guido 

Torzilli, Xiao-Song He, Patrick S. Leung, Guo-Xiang Yang, M. Eric Gershwin, 

Pietro Invernizzi 

603 

Bezafibrate: A novel and effective alternative for relieving 

pruritus in patients with primary biliary cirrhosis 

Anna Reig, Pilar Sese, Albert Pares; Liver Unit, Hospital Clinic, 

University of Barcelona, IDIBAPS, CIBERehd, Barcelona, Spain 

Background and Aims: Pruritus is a common and distressing 

symptom in patients with primary biliary cirrhosis (PBC), and 

when uncontrollable it is an indication for liver transplantation. 

Different therapeutic approaches for pruritus have been 

proposed including resins, rifampicin, naltrexone, sertraline 

and albumin dialysis. Recent observations have indicated that 

fibrates may improve cholestatic itching, although no specific 

studies have been carried out. Therefore, we have assessed 

the effects of fibrates on pruritus in patients PBC who had a 

suboptimal response to ursodeoxycholic acid (UDCA). Patients 

and Methods: 46 PBC patients (43 females, age 54.3 ± 1.5 

years) with suboptimal biochemical response to UDCA were 

treated with bezafibrate (400 mg/d). Apart from clinical and 

biochemical changes, pruritus severity was assessed by a specific 

questionnaires (PBC-40 and pruritus score) and with a 

visual analogue scale (VAS) (form 0 to 10), at baseline and 

after a mean of 29 ± 4 months. Moreover, bezafibrate therapy 

was discontinued in 13 patients to evaluate the course 

of this symptom after bezafibrate withdrawal. Results: Twenty 

seven patients (58.7%) experienced pruritus at baseline (mean 

VAS: 4.4 ± 0.5). No significant differences regarding to prior 

duration of UDCA therapy, age, gender and severity of biochemical 

cholestasis were observed between patients with and 

without pruitus at the beginning of bezafibrate therapy, except 

for ALT which were significantly higher in patients with pruritus. 

Triglyceride and cholesterol levels as well as transient elastography 

(8.9 ± 0.8 vs 9.3 ±2.7 kPa, p:n.s) were also similar in 

patients with and without pruritus. Bezafibrate therapy resulted 

in a significant mitigation of pruritus (VAS from 4.4 ± 0.5 to 

0.8 ± 0.2,


were comparable between PSC, PBC and HC (0.26 [0.37], 

0.23 [0.27] and 0.18 [0.18] ng/mL, resp., p=0.27), baseline 

fasted TBS were elevated in PSC compared to PBC and HC 

(29.0 [62.2], 3.2 [7.0] and 2.8 [3.4] mmol/L, resp., p


nostic groups. To evaluate each analyte as a classifier, receiver 

operating characteristic (ROC) curves were constructed and the 

area under the curve (AUC), sensitivity, and specificity were 

calculated as classifier performance metrics. Multinomial logistic 

regression coupled with a forward stepwise selection procedure 

and a leave-one-out cross validation analysis was then 

used to select analytes for inclusion into composite classifiers. 

RESULTS: Importantly, this methodology is strikingly more sensitive 

and indeed there were 30 glycoconjugates differentially 

expressed between PBC and controls, 27 between PSC and 

controls, and 20 between PBC and PSC. Age contributed to 

the expression of 25 glycoconjugates. Multi-analyte classifiers 

designed as disease-specific (i.e. PBC vs control, PSC vs control, 

and PBC vs PSC) diagnostic tests were able to perform their 

designated tasks perfectly (AUCs, sensitivities and specificities 

of 1). CONCLUSIONS: Glycosylation is significantly altered in 

patients with PBC. Glycoconjugates can serve as sensitive and 

specific biomarkers capable of distinguishing between specific 

autoimmune diseases. Indeed, future work can be used to focus 

not only on diagnosis, but also as biomarkers for disease severity 

and prognosis. 

Disclosures: 

L. Renee Ruhaak - Employment: Intel 




Sciences, Inc 

The following authors have nothing to disclose: Kyoungmi Kim, Sandra L. Taylor, 

Qiuting Hong, Forum Patel, Carol Stroble, Patrick S. Leung, Michiko Shimoda, 

M. Eric Gershwin, Carlito B. Lebrilla, Emanual Maverakis 

607 

Phenotypic Variations in Primary Sclerosing Cholangitis 

Across the Age Spectrum 

John E. Eaton 1 , Brian D. Juran 1 , Elizabeth J. Atkinson 2 , Bryan 

McCauley 2 , Erik M. Schlicht 1 , Mariza de Andrade 2 , Velimir A. 

Luketic 8 , Joseph A. Odin 3 , Ayman A. Koteish 4 , Kris V. Kowdley 5 , 

Kapil B. Chopra 6 , Gideon Hirschfield 7 , Naga P. Chalasani 9 , Konstantinos 

Lazaridis 1 ; 1 Gastroenterology and Hepatology, Mayo 

Clinic, Rochester, MN; 2 Biomedical Statistics and Informatics, 

Mayo Clinic, Rochester, MN; 3 Gastroenterology and Hepatology, 

Icahn School of Medicine at Mount Sinai, New York, NY; 4 Gastroenterology 

and Hepatology, Johns Hopkins, Baltimore, MD; 5 Liver 

Care Network, Swedish Medical Center, Seattle, WA; 6 Gastroenterology 

and Hepatology, University of Pittsburgh, Pittsburgh, PA; 

7 Centre for Liver Research and NIHR Biomedical Research Unit, 

University of Birmingham, Birmingham, United Kingdom; 8 Gastroenterology 

and Hepatology, Virginia Commonwealth University, 

Richmond, VA; 9 Gastroenterology and Hepatology, Indiana University 


Background & Aims: Primary sclerosing cholangitis (PSC) typically 

develops in middle-aged men. However, it is unknown if 

phenotypic differences exist among patients diagnosed with 

PSC at various ages. To this end, we compared the clinical 

characteristics of a PSC cohort based on the age when PSC 

was diagnosed. Methods: We performed a patient survey 

and a medical record review to compare the phenotypic features 

of PSC patients (n=967) who were diagnosed between 

1-19 years (yrs) (early onset, n=108), 20-59 yrs (middle age 

onset, n=763), and 60-79 yrs (late onset, n=96). Patients were 

recruited prospectively from 8 academic medical centers in 

North America. Results: There were no demographic differences 

between groups. However, those with early onset-PSC 

were more likely to have ulcerative colitis (UC) (61.1%) compared 

to the middle age onset (54.1%) and late onset (40.6%) 

groups (p=0.01). Similarly, concomitant autoimmune hepatitis 

was more common in early onset-PSC when compared to 

the other groups (13.9% vs. 4.8% for middle age onset and 

7.3% for late onset, p=0.001). Cholangiocarcinoma (CCA) 

was diagnosed in 54 PSC patients (early onset 0%, middle age 

onset 7% and late onset 8.3%). While CCA in early onset-PSC 

appears to be rare, we did not detect a statistically significant 

difference between groups and the overall development 

of CCA (log rank p value=0.32). However, those with late 

onset-PSC were more likely to be diagnosed with CCA within a 

year of their PSC diagnosis when compared to the other groups 

(early onset 0%, middle age onset 2.5% and late onset 6.2%, 

p = 0.02). Two-hundred and eleven individuals underwent a 

liver transplantation (early onset 14%, middle age onset 25% 

and late onset 6.2%) and survival free from liver transplant was 

similar between groups after adjusting for PSC duration and 

gender (late onset vs. early onset: [Hazard ratio] HR; 2.1, 95% 

Confidence Interval [CI]: 0.8-5.7, p: 0.15; middle age onset 

vs. early onset: HR; 0.8, 95% CI: 0.5-1.5, p=0.50). Lastly, 

early onset-PSC was associated with a higher proportion of 

second-degree relatives with inflammatory bowel disease (IBD) 

when compared to the other groups (19.4% vs. 6.8% for middle 

age onset and 2.1% for late onset, p


cholangitis (ASC) – and 16 healthy subjects (HS) were studied. 

Peripheral blood cell phenotype was determined by flow 

cytometry; ability to suppress was evaluated as inhibition of 

cell proliferation/effector cytokine production; ectoenzymatic 

activity by thin layer chromatography; expression of adenosine 

receptor, adenosine deaminase (ADA) and phosphodiesterases 

(PDE) by quantitative real-time PCR or Western Blot. 

Results: As compared to their CD39 - counterpart, Th17 CD39+ 

cells retained expression of CCR6, IL-23R and RORC, classical 

Th17 cell markers; displayed higher frequencies of CD69 + , 

CD44 + and CD25 + activated cells; contained higher numbers 

of CD73 + , CD161 + and FOXP3 + lymphocytes; and were more 

frequently positive for IL-22, IL-10 and TGF-b producing cells. 

The proportion of Th17 CD39+ cells was markedly lower in AILD 

than HS, with ASC displaying lower proportions than AIH 

patients. In AILD, Th17 CD39+ numbers are greatly decreased 

and have impaired ability to generate AMP/adenosine and 

to control both target cell proliferation and IL-17 production. 

When compared to HS, Th17 cells from AILD patients had a 

markedly decreased expression of A2A adenosine receptor 

while displaying similar expression levels of PDE4A, PDE4B 

and ADA. Conclusions: In AILD, Th17 CD39+ cells are numerically 

decreased and defective in their ability to generate adenosine 

and exert suppression. Failure of these Th17 cells to upregulate 

CD39 appears linked to reduced A2A adenosine receptor levels. 

Low CD39 and A2A expression may contribute, at least 

in part, to the perpetuation of Th17 effector potential in AILD. 

Disclosures: 

Simon C. Robson - Grant/Research Support: Pfizer, NIH, Dainippon; Independent 

Contractor: Biolegend, EMD Millipore, Mersana; Management Position: 

eBioscience; Speaking and Teaching: ACP, Elsevier, ATC; Stock Shareholder: 

Nanopharma, Puretech 

The following authors have nothing to disclose: Rodrigo Liberal, Charlotte R. 

Grant, Yun Ma, Michael A. Heneghan, Giorgina Mieli-Vergani, Diego Vergani, 

Maria Serena Longhi 

OCA on the markers of cholestasis and safety. Study inclusion 

criteria: PBC diagnosis, ALP ≥1.67x ULN and/or total bilirubin 

>ULN to








Karel J. van Erpecum - Advisory Committees or Review Panels: Bristol Meyers 

Squibb, Abbvie, Janssen Cilag, Gilead 

Roya Hooshmand-Rad - Employment: Intercept pharmaceuticals Inc. 

Shawn Sheeron - Employment: Intercept Pharmaceuticals 



The following authors have nothing to disclose: Giuseppe Mazzella, Pietro Invernizzi, 

Joost Drenth, Jaroslaw Regula, Annarosa Floreani, Velimir A. Luketic, 

Victor Vargas, Catherine Vincent, Bettina E. Hansen 

610 

Patients with Primary Biliary Cirrhosis Experience Progressive 

Bone Loss Over Time Beyond that Expected by 

Age and Despite Routine Clinical Care 

Aliya Gulamhusein 1 , Bryan McCauley 2 , Elizabeth J. Atkinson 2 , 

Brian D. Juran 1 , Erik M. Schlicht 1 , Mariza de Andrade 2 , Konstantinos 

Lazaridis 1 ; 1 Center for Basic Research in Digestive Diseases, 

Mayo Clinic, Rochester, Rochester, MN; 2 Health Science 

Research, Mayo Clinic, Rochester, MN 

Background and Aims: Osteopenia and osteoporosis are 

characterized by low bone mass and increased fracture risk. 

Previous reports suggest that patients with primary biliary cirrhosis 

(PBC) are at increased risk of osteoporosis with estimates 

varying between 15-50%, but study populations and testing 

approaches have been heterogeneous. Furthermore, the rate 

of bone loss in these patients is poorly understood. We sought 

to estimate the prevalence of low bone mass and characterize 

the change in bone mineral density (BMD) over time in 

a large cohort of patients with PBC. Methods: We identified 

patients recruited to the Mayo Clinic PBC Genetic Epidemiology 

Registry and Biorepository who had available dual X-ray 

absorptiometry (DXA) scans for assessment of BMD. Those with 

comorbid autoimmune hepatitis were excluded and data was 

censored at the time of liver transplant. Femoral neck (FN) BMD 

(g/cm 2 ) and T-score were used to assess bone loss. Per WHO 

criteria, osteopenia was defined as a T-score of < -1 but > -2.5 

and osteoporosis as a T-score of ≤ -2.5. Linear mixed effects 

models were fit on subjects with multiple scans to estimate the 

changes in BMD over time using gender, age, and time since 

diagnosis as covariates. Results: 383 patients with 885 DXA 

scans were included. There were 813 scans on 348 women 

and 71 scans on 35 men. The median number of scans per 

subject was 2 (25%-75% IQR=1-3) and follow-up was over 9.2 

years (25%-75% IQR=3.4-16.4). The prevalence of low bone 

mass (osteopenia or osteoporosis) in all patients aged


Disclosures: 


Falk, Shire 

The following authors have nothing to disclose: Claire Hardie, Kile J. Green, 

Sarah Pagan, Jessica K. Dyson, Lucy J. Walker, Laura Jopson, John G. Brain 

612 

Vitamin A deficiency promotes hepatic expansion of 

resident (Kupffer cells) and peripheral (Gr hi ) macrophages 

leading to excessive liver damage in rats with 

obstructive cholestasis 

Ali Saeed, Mark Hoekstra, Martijn O. Hoeke, Janette Heegsma, 

Han Moshage, Klaas Nico Faber; Department of Gastroenterology 

and Hepatology, University Medical Center Groningen, Groningen, 

University of Groningen, The Netherlands, Groningen, Netherlands 

Vitamin A deficiency (VAD) is associated with chronic liver diseases 

(CLD), including biliary atresia, primary biliary cirrhosis, 

primary sclerosing cholangitis and non-alcoholic fatty liver disease. 

Serum and/or hepatic retinol levels negatively correlate 

with disease progression. Previously, we found that VAD rats 

shown much more severe liver damage when exposed to bile 

duct ligation (BDL) compared to rats with sufficient vitamin A 

(VAS) levels, but the mechanisms involved remain unresolved 

so far. Vitamin A has potent anti-inflammatory properties and 

therefore we investigated its effect on resident (Kupffer cells) 

and peripheral (Gr hi ) macrophages in vitro and in vivo. Weaning 

male Wister rats were fed either a VAS or a VAD diet for 

14-16 weeks, followed by BDL for 1, 2, 4, 7 days. One group 

subjected to 7-day BDL received retinyl-palmitate therapy. Liver 

damage marker were analysed in serum, and liver tissue was 

analysed with Q-PCR, Western blotting and immunohistochemistry. 

Primary rat Kupffer cells and THP-1 (human Leukemic 

monocyte cell line) cells were exposed to VAS and VAD conditions 

and cytokine expression was analysed by Q-PCR. CD68 

(general macrophage marker) and CD11b (peripheral macrophages) 

positive cells were significantly increased in livers 

of VAD rats after 4- and 7-day BDL as compare to VAS-BDL 

control livers, and was associated with increased plasma γGT 

levels and expression of liver fibrosis makers, such as α-SMA 

and Collagen1A1. All these features were strongly suppressed 

in VAD-BDL rats receiving vitamin A therapy. In line, expression 

of the macrophage chemoattractant Ccl2 was significantly 

increased in VAD-BDL livers and suppressed by vitamin A therapy. 

VAD-BDL livers showed increased expression of proinflammatory 

and profibrotic markers, such as iNos, Tnf-α, Il-1, 

Il-6, Cox-2, Tgf-β and vitamin A therapy decreased this inflammatory 

response. In vitro, VAD (RPMI with lipid-stripped serum) 

caused an inflammatory polarization in primary Kupffer cells 

and PMA-activated-THP-1 cells as compare to control cells and 

increased the expression of inflammatory markers, including 

iNos, Tnf-α, Il-1, Il-6, Cox-2. The vitamin A metabolites, 9-cis 

retinoic acid and all trans retinoic acid, reduced expression 

of these inflammatory markers. In conclusion, vitamin A deficiency 

promotes hepatic macrophage expansion in obstructive 

cholestasis, which show an aggressive pro-inflammatory and 

pro-fibrotic phenotype that aggravates liver damage, but is 

effectively prevented by vitamin A supplementation. This study 

supports an direct role of vitamin A in preventing excessive 

liver damage in patients with chronic liver diseases. 

Disclosures: 

The following authors have nothing to disclose: Ali Saeed, Mark Hoekstra, Martijn 

O. Hoeke, Janette Heegsma, Han Moshage, Klaas Nico Faber 

613 

The Modulation of Co-stimulatory Molecules by Circulating 

Exosomes in Primary Biliary Cirrhosis 

Takashi Tomiyama 1,2 , Guo-Xiang Yang 1 , Ming Zhao 4 , Weici 

Zhang 1 , Hajime Tanaka 1,5 , Jing Wang 4 , Patrick S. Leung 1 , Kazuichi 

Okazaki 2 , Xiao-Song He 1 , Quiajin Lu 4 , Ross L. Coppel 3 , 

Christopher L. Bowlus 1 , M. Eric Gershwin 1 ; 1 Internal Medicine, 

University of California, Davis, CA; 2 Third Department of Internal 

Medicine, Division of Gastroenterology and Hepatology, Kansai 

Medical University, Osaka, Japan; 3 Department of Microbiology, 

Monash University, Melbourne, VIC, Australia; 4 Department of 

Dermatology, The Second Xiangya Hospital, Central South University, 

Hunan, China; 5 Department of Gastroenterology and 

Metabolism, Nagoya City University Graduate School of Medical 

Sciences, Nagoya, Japan 

Background: Exosomes are secreted intracellular microparticles 

that can induce antigen-specific immune responses and potentially 

trigger inflammation. The role of exosomes in autoimmunity 

has drawn significant attention because of their ability to 

modulate cell-to-cell communications in part by their ability to 

regulate cytokine and chemokine activation. However, the contribution 

of exosomes in autoimmune cholangitis has not been 

addressed. Herein, we studied the effect of exosomes from 

patients with primary biliary cirrhosis (PBC) and healthy controls 

(HCs) on cytokine production and co-stimulatory molecule 

expression in peripheral mononuclear cell populations. Further, 

we also determined the micro RNA (miRNA) profiles of circulating 

exosomes and verified with an independent cohort. Methods: 

Exosomes were purified from plasma from 28 patients 

with PBC and 25 HCs. Their effect on cytokine production and 

co-stimulatory molecule expression in mononuclear cell populations 

were examined using an ex vivo system. In addition, miR- 

NAs profile of the exosomes was measured by Agilent Human 

miRNA (8*60K) V19.0 microarray. miRNAs with a level of 

2-fold or larger difference between groups were considered 

differentially expressed and verified using isolated exosomes 

from additional groups of PBC and HC using a Taqman PCR 

assay for individual miRNAs. Further, we examined whether 

the levels of these exosomal miRNAs correlated with severity of 

disease. Results: Our data showed that circulating exosomes 

from PBC are taken up by antigen presenting cells (APCs) and 

affect the expression of cell surface co-stimulatory molecules 

CD80, CD86 and CD40 on APCs. Circulating exosomes in 

PBC contain altered patterns of miRNA expression with 9 miR- 

NAs significantly up-regulated and another 9 miRNAs significantly 

down-regulated when compared to the HC. Conclusions: 

Exosomes significantly alter co-stimulatory molecule expression 

on antigen presenting cell populations and there were differences 

of miRNA expression in circulating exosomes in patients 

with PBC. The rigorous dissection of exosomes and their constituents 

is an important component for analysis to understand the 

nature of the pathogenic effects produced during the natural 

history of autoimmune cholangitis. 

Disclosures: 




Sciences, Inc 

The following authors have nothing to disclose: Takashi Tomiyama, Guo-Xiang 

Yang, Ming Zhao, Weici Zhang, Hajime Tanaka, Jing Wang, Patrick S. Leung, 

Kazuichi Okazaki, Xiao-Song He, Quiajin Lu, Ross L. Coppel, M. Eric Gershwin


614 

Increased expression of ORM1-like protein 3 (ORMDL3) 

in biliary epithelial cells may be related to the pathogenesis 

of primary biliary cirrhosis 

Motoko Sasaki 1 , Yasunori Sato 1 , Yasuni Nakanuma 2,1 ; 1 Human 

Pathology, Kanazawa University Graduate School of Medicine, 

Kanazawa, Japan; 2 Shizuoka Cancer Center, Shizuoka, Japan 

Background/Aims: ORM1-like protein 3 (ORMDL3) localizes 

to the endoplasmic reticulum (ER), mediates calcium homeostasis 

and facilitates the unfolded-protein response, which is 

related to the endogenous induction of inflammation. ORMDL3 

gene is located in chromosome 17q12-q21region, which is 

one of candidates associated with the development of primary 

biliary cirrhosis (PBC), asthma and Crohn’s disease. Although 

upregulated expression of ORMDL3 was reported in airway 

epithelial cells in asthma, the expression of ORMDL3 in biliary 

epithelial cells (BECs) in PBC and other liver diseases has not 

been reported, so far. We hypothesized that altered expression 

of ORMDL3 in BECs in PBC may be related to the deregulated 

autophagy and abnormal expression of mitochondria 

antibodies specifically seen in PBC. Methods: We examined 

immunohistochemically the expression of ORMDL3 in BECs in 

livers taken from patients with PBC (n=50), control diseased 

livers (n=55), such as primary sclerosing cholangitis (PSC), 

and normal livers (n=20). The association between the expression 

of ORMDL3 and the expression of mitochondrial antigen 

PDC-E2, autophagy-related markers (LC3, p62) or senescent 

markers (p16 INK4a and p21 WAF1/Cip1 ) was also examined. In 

addition, a correlation between the extent of biliary expression 

of ORMDL3 and the extent of ORMDL3-positive inflammatory 

cells infiltration in small bile ducts was evaluated. Furthermore, 

we examined the expression of ORMDL3 at mRNA level in cultured 

BECs treated with various cytokines (IL-4 [10ng/ml]; IL-13 

[10ng/ml]; IFN-γ [1000U/ml], TNF-α [10ng/ml] and TGF-β 

[4ng/ml]) and a protein kinase A activator, forskolin (5 mM). 

Results: The expression of ORMDL3 was seen in the cytoplasm 

and apical surface in damaged small bile ducts (SBDs) in PBC. 

The expression of ORMDL3 was significantly more extensive 

in SBDs in PBC, especially in early stage PBC, compared with 

control diseased livers and normal livers (p


616 

Association between elevated serum IgG4 (sIgG4) concentrations 

and the phenotype of patients with primary 

sclerosing cholangitis (PSC) 

Michael P. Manns 1 , Bertus Eksteen 2 , Mitchell L. Shiffman 3 , Cynthia 

Levy 4 , Kris V. Kowdley 5 , Aldo J. Montano-Loza 6 , Harry L. Janssen 

7 , Robert P. Myers 8 , Dora Ding 8 , Mani Subramanian 8 , John 

G. McHutchison 8 , Michael R. Charlton 9 , Christopher L. Bowlus 10 , 

Andrew J. Muir 11 , Roger W. Chapman 12 ; 1 Hannover Medical 

School, Hannover, Germany; 2 University of Calgary, Calgary, AB, 

Canada; 3 Liver Institute of Virginia, Richmond, VA; 4 University of 

Miami, Miami, FL; 5 Swedish Medical Center, Seattle, WA; 6 University 

of Alberta, Edmonton, AB, Canada; 7 University of Toronto, 

Toronoto, ON, Canada; 8 Gilead Sciences, Inc., Foster City, CA; 

9 Intermountain Medical Center, Salt Lake City, UT; 10 University of 

California at Davis, Sacramento, CA; 11 Duke Clinical Research 

Institute, Durham, NC; 12 University of Oxford, Oxford, United 

Kingdom 

Background: Elevated sIgG4 has been reported in 9-15% of 

PSC patients and is associated with a more aggressive disease 

course. Our aim was to compare the characteristics 

of PSC patients with elevated and normal sIgG4. Methods: 

We measured sIgG4 (BN II System; Siemens, Malvern, PA) 

in PSC patients enrolled in a phase 2b trial of simtuzumab. 

Corticosteroid and/or anti-TNF-α therapies were prohibited. 

The associations between elevated sIgG4 (>140 mg/dL) with 

demographics, body mass index (BMI), ulcerative colitis (UC), 

use of ursodeoxycholic acid (UDCA), liver biochemistry, Fibro- 

Test, ELF, sLOXL2 (VIDAS® LOXL2; bioMérieux, Marcy L’Etoile, 

France), liver fibrosis staged by the Ishak classification, and 

Mayo risk score (MRS) were determined. MRCP data will be 

available at the time of presentation. Results: Among 234 

patients, 34 (14.5%) had elevated sIgG4. These patients were 

older than those with normal sIgG4, but sex, race, UC, and use 

of UDCA did not differ between groups (Table). Although liver 

biochemistry did not differ, patients with elevated sIgG4 had 

lower serum albumin and higher platelet levels compared with 

those with normal sIgG4. FibroTest, ELF and sLOXL2, the proportion 

of patients with bridging fibrosis or cirrhosis, and MELD 

and MRS were similar between groups. A sensitivity analysis 

examining a sIgG4 cut-off of >201 mg/dL revealed similar 

findings. Conclusions: A small proportion of PSC patients have 

elevated sIgG4. In this clinical trial cohort, the sIgG4 level 

does not have a significant impact on PSC phenotype including 

disease severity assessed biochemically, histologically, or 

according to conventional prognostic indices. 

Characteristics of PSC Patients According to sIgG4 

Median (IQR) or % (n). 

Disclosures: 





Novartis 










Gilead 






Robert P. Myers - Employment: Gilead Sciences, Inc.; Stock Shareholder: Gilead 




Sciences 






Sciences, Inc 




The following authors have nothing to disclose: Bertus Eksteen, Cynthia Levy, 

Aldo J. Montano-Loza, Dora Ding, Roger W. Chapman 

617 

Modeling biliary fluid dynamics reveals possible mechanism 

for dose-response and personalization of UDCA 

treatment in PSC 

Oleksandr Ostrenko 1 , Fabian Segovia-Miranda 2 , Mario Brosch 6 , 

Wiebke Erhart 5 , Kirstin Meyer 2 , Georg Kretzschmar 3 , Christoph 

Jüngst 4 , Frank Lammert 4 , Marino Zerial 2 , Clemens Schafmayer 5 , 

Lutz Brusch 1 , Jochen Hampe 6 ; 1 Center for Information Services 

and High Performance Computing, Technische Universität Dresden, 

Dresden, Germany; 2 Max Planck Institute of Molecular Cell 

Biology and Genetics, Dresden, Germany; 3 Molecular Cell Physiology 

and Endocrinology, Institute of Zoology, Technische Universität 

Dresden, Dresden, Germany; 4 Saarland University Medical 

Center, Homburg, Germany; 5 University Hospital Schleswig Holstein, 

Kiel, Germany; 6 TU Dresden, University Hospital Dresden, 

Dresden, Germany 

Background: Primary sclerosing cholangitis (PSC) is a progressive 

liver disease characterized by fibroobliterative destruction 

of the intra- and/or extra-hepatic bile ducts. Combined with 

immune-mediated insults to the bile duct, biliary flow obstruction 

might lead to pressure damage to the biliary epithelium 

and may drive further disease progression. Currently, the only 

and controversial medical treatment is the choleretic drug 

ursodeoxycholic acid (UDCA). However, while moderate doses 

(10-15mg/kg/day) of UDCA might improve liver function tests 

and histology, high-dose UDCA has been demonstrated to 

increase mortality in a RCT, thereby calling UDCA treatment 

in question. Aims: Develop a hydrodynamic model of biliary 

pressure and flow to assess the effect of UDCA on biliary pressure 

in normal liver and PSC. Methods: A recently developed 

theory of bile secretion and transport was applied to UDCA 

treatment. UDCA reduces bile viscosity (1) and increases solute 

concentration and consequently osmotic water influx into bile


(2). Model parametrization was completed using measured 

viscosity from gallbladder bile of patients treated with UDCA 

and quantitative 3D-reconstruction of stacked high resolution 

microscopy of healthy human and PSC livers. Bile viscosity and 

osmotic water inflow are modelled as functions of UDCA dose 

according to effects (1) and (2) above. Results: This quantitative 

model allows the assessment of the mechanical consequences 

of UDCA treatment in a dose-dependent manner and predicts 

intrahepatic fluid pressure as a function of the physical properties 

of bile and the geometrical properties of the tissue. For 

healthy liver, the model reproduces the insensitivity of biliary 

pressure to UDCA dose. In PSC, a pronounced dose-response 

curve is predicted by our model: biliary pressure is decreasing 

at UDCA dose increments, reaches a minimum and then rapidly 

increases beyond baseline upon further dose increments. The 

optimal UDCA dose corresponding to the maximum pressure 

reduction can be quantified as a function of the physical and 

geometrical parameters that may vary from patient to patient. 

Conclusions: This study provides a hydrodynamic explanation 

of the clinical dose-response of UDCA in PSC. Further, it confirms 

tolerance to high doses of the compound in normal liver. 

Mechanistically, the UDCA-induced increase in water inflow 

has pressure-lowering (bile dilution) and pressure-increasing 

(increased fluid volume in need of drainage) consequences. 

Thus, the model advocates individualized dose-optimization 

based on the patient-specific biliary micro-geometry, thereby 

potentially rendering UDCA treatment safer and more widely 

applicable. 

Disclosures: 

The following authors have nothing to disclose: Oleksandr Ostrenko, Fabian 

Segovia-Miranda, Mario Brosch, Wiebke Erhart, Kirstin Meyer, Georg Kretzschmar, 

Christoph Jüngst, Frank Lammert, Marino Zerial, Clemens Schafmayer, Lutz 

Brusch, Jochen Hampe 

618 

In PSC with dominant bile duct stenosis, multi-resistant 

bacteriobilia is associated with reduced survival 

Christian Rupp 1 , Hannah Salzer 1 , Konrad A. Bode 2 , Karl Heinz 

Weiss 1 , Wolfgang Stremmel 1 , Peter Sauer 1 , Daniel Gotthardt 1 ; 

1 Internal Medicine IV, University Hopsital Heidelberg, Heidelberg, 

Germany; 2 Department of Infectious Diseases, University of Heidelberg, 

Heidelberg, Germany 

Background & Aims: Primary sclerosing cholangitis (PSC) is 

a chronic cholestatic liver disease, characterized by sclerosis 

and destruction of the biliary system. The course of diseases is 

often complicated by biliary infections. We aimed to analyze 

the frequency and influence of cholangitis with multi-resistant 

bacteria in PSC patients. Methods: Patients who were admitted 

to our Department during the time period of 1987 and 

2014 with well-defined diagnosis of PSC were analyzed in 

regard to multi-resistant bacteria in bile cultures. Comparison 

between frequencies was done using Chi-Square-test or Fisher’s 

exact test where appropriate. Continuous data were compared 

with the nonparametric Wilcoxon rank-sum test. Actuarial 

transplantation-free survival was estimated using Kaplan-Meier 

product limit estimator. Differences between the actuarial estimates 

were tested with the log rank test. Results: We identified 

20/244 (8.3) patients with multi-resistant bacteriobilia. 

8 patients had vancomycin resistant enterococcus (VRE) and 

12 had multi-resistant gram negative bacteria (MRGN) in bile 

culture. Baseline characteristic including laboratory values 

(AST, ALT, GGT, AP, Bilirubin), age at onset of PSC, concomitant 

inflammatory bowel disease, overlap with autoimmune 

hepatitis, dominant stenosis and Mayo Risk Score were not 

different between patients with or without MR bacteria. There 

was no difference in frequency of MR bacteriobilia between 

patients with or without DS (14/159, 8.8% vs. 5/66, 7.6%; 

p=0.8). We performed Kaplan-Meier analysis showing a markedly 

reduced transplantation-free survival in PSC patients with 

MR bacteriobilia (17.2 vs. 11.0 years; p=0.014). Stratification 

for presence of DS revealed reduced survival in presence 

of MR bacteriobilia only in patients with DS (16.7 vs. 9.9 

years; p=0.001), whereas no influence was detectable in PSC 

patients without DS (21.0 vs. 17.8 years; p=0.8). In multivariate 

analysis, including gender, age, AIHOL, IBD, DS, number 

of endoscopic interventions, MRS and MR bacteriobilia, only 

IBD (HR 3.3, 95% CI 1.1-11.1; p=0.04), MRS (HR 1.5, 95% 

CI 1.1-2.1; p=0.04) and MR bacteriobilia (HR 3.2, 95% CI 

1.1-9.5; p=0.03) were independent risk factors for reduced 

transplantation-free survival. Conclusion: In PSC patients with 

dominant stenosis MR bacteriobilia is associated with reduced 

survival, independent of number of endoscopic interventions. 

Multi-resistant bacteria may play a role in the progression of 

PSC. Optimal treatment strategies need to be established in 

order to achieve eradication of MR bacteriobilia. 

Disclosures: 

The following authors have nothing to disclose: Christian Rupp, Hannah Salzer, 

Konrad A. Bode, Karl Heinz Weiss, Wolfgang Stremmel, Peter Sauer, Daniel 

Gotthardt 

619 

Innate Immunity Drives the Initiation of Autoimmune 

Cholangitis in a Xenobiotic Murine Model of Primary 

Biliary Cirrhosis 

Chao-Hsuan Chang 2 , Ying-chun Chen 2 , Weici Zhang 1 , Patrick S. 

Leung 1 , M. Eric Gershwin 1 , Ya-Hui Chuang 2 ; 1 Internal Medicine, 

University of California, Davis, CA; 2 Department of Clinical Laboratory 

Sciences and Medical Biotechnology, National Taiwan 

University, Taipei, Taiwan 

Background: Invariant natural killer T (iNKT) cells are important 

in bridging innate and adaptive immunity by engaging 

with glycolipids presented by CD1d. α-galactosylceramide 

(α-GalCer) is a specific ligand for iNKT cell activation. Modification 

of α-GalCer lipid chain results in the generation of 

glycolipids with predominant Th1 or Th2 cytokine skewing 

profiles. (2s,3s,4r)-1-O-(α- D 

-galactopyranosyl)-N-tetracosanoyl-2-amino-1,3,4-nonanetriol 

(OCH) is a synthetic analog 

of α-GalCer which stimulates iNKT cells to predominantly Th2. 

Previous work in our group has demonstrated that iNKT cell 

activation by α-GalCer resulted in profound exacerbation of 

portal inflammation, bile duct damage, and hepatic fibrosis 

in the 2 octynoate-BSA (2OA-BSA) xenobiotic murine model 

of PBC. Here, we examined whether iNKT cells activated by 

a Th2-biasing agonist OCH, can influence the development of 

PBC in 2OA-BSA immunized mice. Methods: Groups of mice 

were treated with either OCH or α-GalCer and serially followed 

for cytokine production, markers of T cell activation, 

liver histopathology and anti-mitochondrial antibodies (AMA). 

In addition, groups of CD1d deleted mice and wild type mice 

were immunized with 2OA-BSA and monitored for liver infiltrates 

including CD4+ T cells, CD8+ T cells, NKT, NK and B 

cells. IFN-γ production by liver mononuclear cells were also 

compared. Results: 2OA-BSA mice treated with OCH exhibited 

a Th2 biased cytokine; α-GalCer initiated a rapid IL-4 

and prolonged IFN-γ production, whereas OCH induced predominant 

IL-4 production. Both OCH and α-GalCer induced 

high levels of AMA in 2OA-BSA immunized mice and had 

exacerbated portal inflammation and hepatic fibrosis. However, 

at 12 weeks post immunization, 2-OA-BSA/OCH immunized 

mice had significantly higher liver total mononuclear 

cell infiltrates, increased numbers of T, B and NK cells, and


increased CD4 + and CD8 + T cells compared to 2-OA-BSA/PBS 

immunized mice. The frequencies of CD44 expressing CD8 + 

T cells and CD69 expressing CD8 + T cells were significantly 

increased in 2-OA-BSA/OCH immunized mice compared to 

2-OA-BSA/PBS immunized mice. Moreover, the frequency of 

CD44 expressing CD4 + T cells was significantly increased in 

2-OA-BSA/OCH immunized mice. Furthermore, the levels of 

AMA, cell infiltrates, and IFN-γ production of liver mononuclear 

cells were decreased in CD1d -/- mice when compared with 

wild type mice. Conclusion: Activation of iNKT cells is critical 

for the natural history of autoimmune cholangitis in this model 

and highlights the role of innate immunity in human PBC. 

Disclosures: 

The following authors have nothing to disclose: Chao-Hsuan Chang, Ying-chun 

Chen, Weici Zhang, Patrick S. Leung, M. Eric Gershwin, Ya-Hui Chuang 

620 

Serum metabolomic profile of patients with primary 

biliary cirrhosis treated with bezafibrate and changes 

following itching relief 

Albert Pares 1 , Anna Reig 1 , Miriam Perez-Cormenzana 2 , Rebeca 

Mayo 2 , Pilar Sese 1 , Azucena Castro 2 ; 1 Liver Unit, Hospital Clinic, 

University of Barcelona, IDIBAPS, CIBERehd, Barcelona, Spain; 

2 OWL, Derio, Spain 

Background and aims: Long-term fibrate treatment is an effective 

therapy for patients with primary biliary cirrhosis with 

suboptimal biochemical response to ursodeoxycholic acid. 

Bezafibrate therapy has further effects, thus decreasing pruritus 

in this cholestatic condition. The metabolomic consequences 

of fibrates in PBC are not known, and some studies indicate 

that their action may result from being a PPAR alpha agonist 

and also increasing the expression of some bile acid transporters. 

Therefore, we have assessed the metabolomic profiling of 

patients under bezafibrate, and particularly in those with pruritus, 

to define the potential mechanisms of action of bezafibrate 

in this cholestatic disease. Patients and Methods: Serum samples 

before and after bezafibrate therapy were taken from 29 

patients with PBC. Moreover in 14 of these patients with pruritus, 

samples were obtained before, during and after bezafibrate 

discontinuation. Metabolite extraction was accomplished 

by fractionating the samples into pools of species with similar 

physicochemical properties. Three different UPLC-MS analytical 

platforms were used for the analysis of fatty acyls, bile acids, 

steroids and lysoglycerophospholipids; amino acids; glycerolipids, 

glycerophospholipids, sterol lipids and sphingolipids. 

Results: More than 530 metabolites were identified. Regarding 

individual species 93 metabolites changed during bezafibrate 

therapy: PE(0:0/16:1) and ChoE(16:1) were in higher concentrations, 

while PC(17:0/18:2), PC(17:0/0:0), PC(15:0/22:6), 

PC(18:3/18:3) and PC(18:0/20:4) decreased with bezafibrate. 

Pruritus disappeared or was minimized by bezafibrate. 

In these patients 38 metabolites decreased (p


iary expression of SCT/SR/CFTR/AE2 compared to WT. SCT 

levels in biliary supernatants was decreased, but increased in 

serum in dnTGFβRII mice compared to WT, suggesting that SCT 

comes from other sources such as S cells. There was increased 

fibrosis and SCT/SR/CFTR/AE2 in early stage PBC samples, 

but decreased expression of SCT/SR/CFTR/AE2 in advanced 

PBC samples compared to controls. SCT levels increased in 

serum but decreased in bile in PBC compared to control. Conclusion: 

Loss of biliary bicarbonate secretion may contribute 

to disease progression, which may be partly counteracted by 

peripheral sources of SCT. 

Disclosures: 

The following authors have nothing to disclose: Lindsey Kennedy, Shannon S. 

Glaser, Francesca Bernuzzi, Fanyin Meng, Julie Venter, Heather L. Francis, 

Domenico Alvaro, M. Eric Gershwin, Antonio Franchitto, Paolo Onori, Sharon 

DeMorrow, Marco Marzioni, Eugenio Gaudio, Pietro Invernizzi, Gianfranco 

Alpini 

622 

Colectomy is associated with development of hepatobiliary 

malignancy but not cholangiocarcinoma in patients 

with Primary Sclerosing Cholangitis. 

Kate D. Williamson 1,2 , Ladislav Kozak 3 , Said Al Mamari 1 , John 

Halliday 1,2 , Roger W. Chapman 1,2 ; 1 Translational Gastroenterology 

Unit, John Radcliffe Hospital, Oxford, United Kingdom; 2 Nuffield 

Department of Medicine, University of Oxford, Oxford, United 

Kingdom; 3 Department of Sociology, University of Oxford, Oxford, 


Introduction: Primary Sclerosing Cholangitis (PSC) is associated 

with increased risk of malignancy – including hepatobiliary 

malignancies and colorectal cancer. Colectomy is often 

performed in patients due to either active inflammatory bowel 

disease (IBD) or development of dysplasia or colorectal cancer. 

Recently, the Mayo Clinic in US presented an abstract demonstrating 

an association between colectomy in PSC patients and 

the development of cholangiocarcinoma (CCA). We sought to 

validate this association. Methods: A database of 272 patients 

with PSC exists at John Radcliffe Hospital, Oxford UK, containing 

information on baseline demographics and clinical course. 

We performed a Cox regression analysis to identify variables 

associated with development of firstly CCA, and secondly any 

hepatobiliary malignancy (hepatocellular cancer, gallbladder 

cancer, other cancers arising within the liver, and cholangiocarcinoma). 

Variables examined were age at PSC diagnosis, 

gender, presence of IBD, and whether the patient underwent 

colectomy (including whether the indication was for active disease 

or for a composite of dysplasia or colorectal cancer). 

Results: Data were available for 228 patients. Median age at 

PSC diagnosis was 46.3 years (12.5 – 83.2y), and 61% were 

male. 4/228 (1.8%) had PSC/overlap with autoimmune hepatitis 

and were included in the analysis. The median follow up 

was 8.9 years (0.1 – 29.9y). 13/228 (5.7%) had a diagnosis 

of CCA, whilst 6 had a diagnosis of an alternative hepatobiliary 

malignancy, hence 19/228 (8.3%) had a diagnosis of any 

hepatobiliary malignancy. 18/19 patients with any hepatobiliary 

malignancy diagnosis had died at time of last follow up. 

18% patients had undergone a colectomy, the indication being 

dysplasia or cancer in 15% cases. On multivariate Cox proportional 

hazard models, presence of IBD, age at PSC diagnosis 

and gender were not associated with development of CCA, 

nor the composite endpoint of any hepatobiliary malignancy. 

Likewise, colectomy was not associated with development of 

CCA (HR 0.71, 95%CI 0.15-3.35, p=ns). However, colectomy 

was associated with development of any hepatobiliary 

malignancy (HR of 70.39, 95% CI 4.00-1238.52, p


studies in PSC. Furthermore, ALP may be an important factor to 

include in prognostic modeling for PSC. 

Disclosures: 







The following authors have nothing to disclose: Elisabeth M. de Vries, Junfeng 

Wang, Mariska M. Leeflang, Kirsten Boonstra, Ronald Geskus 

Diagnostic Performance of Serum Fibrosis Markers in PSC 

624 

Validation of serum fibrosis marker panels in patients 

with primary sclerosing cholangitis (PSC) in a randomized 

trial of simtuzumab 

Christopher L. Bowlus 1 , Keyur Patel 2 , Indra Neil Guha 3 , Roger W. 

Chapman 4 , Olivier Chazouillères 5 , Naga P. Chalasani 6 , John M. 

Vierling 7 , Robert P. Myers 8 , Dora Ding 8 , Raul E. Aguilar Schall 8 , 

Mani Subramanian 8 , John G. McHutchison 8 , Andrew J. Muir 2 , 

Zachary D. Goodman 9 , Cynthia Levy 10 ; 1 University of California at 

Davis, Sacramento, CA; 2 Duke Clinical Research Institute, Durham, 

NC; 3 NIHR Nottingham Digestive Diseases Biomedical Research 

Unit, Nottingham, United Kingdom; 4 Oxford University, Oxford, 

United Kingdom; 5 Hôpital Saint Antoine, Paris, France; 6 Indiana 

University, Indianapolis, IN; 7 Baylor College of Medicine, Houston, 

TX; 8 Gilead Sciences, Inc., Foster City, CA; 9 Inova Fairfax 

Hospital, Falls Church, VA; 10 University of Miami, Miami, FL 

Background: Our objective was to assess the diagnostic performance 

of serum fibrosis marker panels in patients with PSC. 

Methods: We calculated FibroTest, ELF, APRI, and FIB-4 and 

measured serum LOXL2 (sLOXL2) by immunoassay (VIDAS® 

LOXL2; bioMérieux, Marcy L’Etoile, France) in PSC patients 

enrolled in a phase 2b trial of simtuzumab. Liver fibrosis was 

staged according to the Ishak classification and their diagnostic 

performance for predicting bridging fibrosis (Ishak stages 

3-6 vs. 0-2) and cirrhosis (stages 5-6 vs. 0-4) was determined 

using AUROCs. Sensitivity analyses were conducted according 

to biopsy length (≥ vs.


trolled studies (2 Phase 2 trials and 1 Phase 3). Abnormal 

bilirubin was not a common feature in the individual studies, 

therefore, data from all three studies was pooled; since all 

studies included a 3-month point, integrated data was evaluated 

at 3 months. ALP was markedly elevated at baseline 

in patients with abnormal bilirubin and OCA treatment was 

associated with significant ALP reductions. Mean bilirubin levels 

increased in the placebo arm and decreased with OCA, 

although the difference was not statistically significant over 

this short 3-month timeframe. Further evaluation of a subset of 

subjects treated to 12 months showed a significant reduction 

vs. placebo (Mean±SE:-8.9±2.2 vs -0.7±1.7; p


28d with no exclusions for severity or treatment of PRU. Patients 

were stratified by presence of prurius (PRU) at BL using the 5-D 

Itch Scale (5D). PRU was measured by Visual Analogue Scale 

(VAS) and the 5D at BL and Days 14, 28 and 42. Subjects 

with a BL and Day 28 VAS (n=38) and/or 5D (n=41) were 

categorized as improved, no change or worsened based on a 

>20% change from BL at Day 28. Correlations between PRU 

and liver chemistries or serum BAs by VAS were assessed by 

Spearman correlation and regression analyses with a clinical 

anchor threshold of r >0.30 and p


Disclosures: 




Gilead 

Hemant Shah - Advisory Committees or Review Panels: Boehringer-Ingelheim; 

Consulting: Hoffman La-Roche, Merck, Gilead; Speaking and Teaching: Vertex 









Roya Hooshmand-Rad - Employment: Intercept pharmaceuticals Inc. 

The following authors have nothing to disclose: Velimir A. Luketic 

629 

Thyroid dysfunction in primary biliary cirrhosis: a comparative 

study at two European centers. 

Chiara Mangini 1 , Ana Reig 2 , Irene Franceschet 1 , Nora Cazzagon 

1 , Lisa Perini 1 , Llorenç Caballeria 2 , Silvia Cocchio 3 , Vincenzo 

Baldo 3 , Albert Pares 2 , Annarosa Floreani 1 ; 1 Department of Surgery, 

Oncology and Gastroenterology, University of Padova, 

Padova, Italy; 2 Liver Unit, Hospital Clinic-IDIBAPS, University of 

Barcelona, Barcelona, Spain; 3 Department of Molecular Medicine, 

Laboratory of Public Health and Population Studies, University of 

Padova, Padova, Italy 

Primary Biliary Cirrhosis (PBC) is often associated with other 

autoimmune diseases, but there are few data about the influence 

of other diseases on the natural history of PBC. Thyroid 

diseases have mainly an autoimmune aetiology, are often diagnosed 

in women and can be associated with PBC. Aim: To 

analyze the association between PBC and thyroid diseases 

and the impact of these disorders on the natural history of 

PBC in two European centres. Methods: 921 PBC patients 

enrolled between 1969 and 2015 in Padova (376 patients, 

M:F=25:351, mean follow-up 115.2 ± 85.6 months) and Barcelona 

(545 patients, M:F=42:503, mean follow-up 135.0 

± 94.9 months) were considered. Data on histological stage 

at diagnosis, biochemical data, associated thyroid disorders, 

extrahepatic autoimmune conditions, clinical events including 

hepatic decompensation (ascites, spontaneous bacterial 

peritonitis, encephalopathy, variceal bleeding and HCC 

development) were recorded. Survival was analysed using 

Kaplan-Meier curves and Cox regression method. Results: 150 

patients (16.3 %) had a thyroid disorder: 94 patients (10.2 %) 

had Hashimoto’s thyroiditis, 15 (1.6 %) had Graves’ disease, 

22 (2.4 %) had a multi-nodular goitre, 7 (0.8 %) had thyroid 

cancer, 13 (1.4 %) had other thyroid disorders. The prevalence 

of different types of thyroid disease was similar in Padova and 

Barcelona, except for Graves’ thyroiditis and thyroid cancer 

that resulted more frequent in the Padova cohort (15.7% vs 

5.0% and 8.6% vs 1.3% respectively, p


64% with an average intensity of 1.7 + 0.9 (scale 1-4). Over 

90% of participants were willing to donate biospecimens and 

be contacted for clinical research. Conclusions: Participants in 

the PSC Partners Registry are similar to other cohorts but are 

majority female, more symptomatic, and are a resource for 

future research. 

for PBC, a clear and important unmet need persists, as corroborated 

by variable UDCA response. The ongoing unmet need 

for PSC is striking, despite widespread UDCA use, and in the 

face of rising rates of transplantation for other liver diseases. 

Table 1. Comparison of Confirmed versus Unconfirmed PSC 

Cases 

Disclosures: 

Gregory T. Everson - Advisory Committees or Review Panels: Roche/Genentech, 

Abbvie, Galectin, Boehringer-Ingelheim, Eisai, Bristol-Myers Squibb, HepC 

Connection, BioTest, Gilead, Merck; Board Membership: HepQuant LLC, PSC 

Partners, HepQuant LLC; Consulting: Abbvie, BMS, Gilead, Bristol-Myers Squibb; 

Grant/Research Support: Roche/Genentech, Pharmassett, Vertex, Abbvie, Bristol-Myers 

Squibb, Merck, Eisai, Conatus, PSC Partners, Vertex, Tibotec, GlobeImmune, 

Pfizer, Gilead; Management Position: HepQuant LLC, HepQuant LLC; 

Patent Held/Filed: Univ of Colorado; Speaking and Teaching: Abbvie, Gilead 




Sciences, Inc 

The following authors have nothing to disclose: Rachel Gomel, Ricky Safer, Jesse 

A. King, Estella M. Geraghty, Keith D. Lindor 

631 

Cholestatic liver disease and liver transplantation in the 

UK: a twenty year review 

Gwilym Webb 1,2 , James W. Ferguson 2 , David Jones 4 , James Neuberger 

2,3 , Gideon Hirschfield 1,2 ; 1 NIHR Centre for Liver Research, 

University of Birmingham, Birmingham, United Kingdom; 2 Hepatology, 

Queen Elizabeth Hospital, Birmingham, United Kingdom; 

3 NHS Blood and Transplant, Bristol, United Kingdom; 4 Newcastle 

University, Newcastle, United Kingdom 

Background: Primary biliary cirrhosis (PBC) and primary sclerosing 

cholangitis (PSC) represent major indications for orthotopic 

liver transplantation (OLT). Ursodeoxycholic acid (UDCA) has 

been prescribed in the UK for most patients with PBC and PSC, 

with consensus for efficacy only in PBC. Aim: To analyse transplant 

listing and transplantation for PBC and PSC over the last 

20 years of UK-wide practice. Results: We analysed national 

transplant registry data for 1995 to 2014 inclusive. Records for 

12,935 listings and 10,512 OLTs were reviewed. Listings for 

all indications increased from 494 to 976/year (21.2±2.3/ 

year; r 2 =0.82; p


were immunoreactive for both markers with a wide range of 

expression (MMP9: median HS, 11.6; range 2.4-122; and 

p-P38: median HS, 54; range 0.9-185). The most intense 

MMP9 expression was in areas of portal inflammation and 

interface hepatitis where immunoreactivity was associated with 

macrophages or within the extracellular matrix. Also, random 

lobular macrophages and Kupffer cells were strongly MMP9 

positive irrespective of the extent of portal inflammation. MMP9 

expression was associated with bile canalicular cholestasis 

(P=0.001) and negatively with periductal fibrosis (P=0.03), but 

was not associated with Ishak stage, the grade of portal, lobular 

or interface inflammation, duct injury, CK7 staining, liver 

biochemistry, or UC. Within inflamed portal areas, nuclear 

p-P38 localized to small mononuclear cells and less frequently 

to reactive duct epithelial cells. p-P38 expression was not associated 

with any histologic feature, liver biochemistry, or UC. 

Conclusions: In PSC patients, hepatic MMP9 and p-P38 expression 

is increased. Additional study is necessary to validate 

that MMP9 and ASK1 pathway activation may contribute to 

disease pathogenesis. 

Disclosures: 

Dorothy French - Employment: Gilead Sciences; Stock Shareholder: Genentech 

- Roche 

Zachary D. Goodman - Consulting: Gilead Sciences, Abbvie; Grant/Research 

Support: Gilead Sciences, Fibrogen, Galectin Therapeutics, Intercept, Synageva, 

Conatus, Tobira, Exalenz 

Erik G. Huntzicker - Employment: Gilead Sciences; Stock Shareholder: Gilead 

Sciences 

Satyajit Karnik - Consulting: Monsoon Dx; Employment: Gilead Sciences 

Victoria Smith - Employment: Gilead Sciences Inc 

Raul E. Aguilar Schall - Employment: Gilead Sciences, Inc. 

Bittoo Kanwar - Employment: Gilead Sciences 



Sciences 






The following authors have nothing to disclose: David Newstrom 

to the modified Amsterdam score (mAm score). Results: In total 

154 PSC patients with adequate bile sample were included. 

The mean age of patients was 40.9±12.7 years. The median 

(IQR) disease duration was 4 (3 - 9) years. Concomitant IBD 

was present at 71 %. The mean (±SD) S-ALP was 137±123 

IU/l (≤ 105 IU/l), S-bilirubin 14.6±12.8mmol/l, (≤ 20 IU/l. 

The mean UDCA dose was 17.1±4.4 mg/kg/day. UDCA 

therapy was associated with markedly increased proportion 

of UDCA in bile (49.1± 22.2 %). The presence or absence 

of IBD had no impact on biliary UDCA content. S-ALP IU/ml 

correlated positively with mAm score, p for linearity 0.005, but 

S-ALP could be < UNL even in advanced biliary disease. No 

correlation was found between UDCA dose and S-ALP level, 

r=0.13 (95%CI: -0.07-0.27). S- ALP level did not correlate with 

the UDCA content (mg%) in bile, fig 1. Conclusions. S-alkaline 

phosphatase in not an optional surrogate marker for disease 

progression or monitoring the treatment response of UDCA in 

PSC patients. 

Figure 1. 

Disclosures: 

The following authors have nothing to disclose: Martti A. Färkkilä, Kalle Jokelainen, 

Hannu Kautiainen 

633 

S-Alkaline phosphatase is not a reliable surrogate 

marker for ursodeoxycholic acid therapy in patients 

with primary sclerosing cholangitis 

Martti A. Färkkilä 1,2 , Kalle Jokelainen 2 , Hannu Kautiainen 1 ; 1 Helsinki 

University, Helsinki, Finland; 2 Gastroenterology, Helsinki University 

Hospital, Helsinki, Finland 

Background. Primary sclerosing cholangitis (PSC) is chronic 

cholestatic liver disease of unknown origin, leading to inflammation 

and strictures of intra- and extrahepatic bile ducts. 

Ursodeoxycholic acid (UDCA is widely used to treat PSC, 

demonstrating amelioration of cholestatic liver enzymes. S-ALP 

is the most used surrogate marker for treatment response for 

UDCA therapy. Objectives of the study.To analyze the effect 

of UDCA therapy 20mg/kg/day on biliary UDCA content and 

S-ALP-levels in PSC. Patients and methods. The study population 

consists of PSC patients on UDCA therapy (n= 154) and 

referred for ERCP for confirmation of the diagnosis of PSC or 

for biliary dysplasia surveillance. After cannulation the common 

bile duct a bile sample was aspirated using balloon catheter, 

immersed immediately in liquid nitrogen and stored in -20 

o C. The content of UDCA in bile was analyzed. The conten of 

UDCA was expressed as molar percentages (mM%) of total 

bile acids. Cholangiographic findings were scored according


634 

Elevation of alkaline phosphatase during follow-up is 

an early predictor of hyperbilirubinaemia and of clinical 

endpoints in primary biliary cirrhosis – an international 

study 

Willem J. Lammers 1 , Henk R. van Buuren 1 , Cyriel Y. Ponsioen 2 , 

Harry L. Janssen 3 , Annarosa Floreani 4 , Gideon Hirschfield 5 , 

Christophe Corpechot 6 , Marlyn J. Mayo 7 , Pietro Invernizzi 8 , 

Pier Maria Battezzati 9 , Albert Pares 10 , Frederik Nevens 11 , Douglas 

Thorburn 12 , Andrew Mason 13 , Kris V. Kowdley 14 , Angela 

C. Cheung 3 , Teru Kumagi 15 , Palak J. Trivedi 5 , Raoul Poupon 6 , 

Ana Lleo 8 , Llorenç Caballeria 10 , Keith D. Lindor 16,17 , Maren H. 

Harms 1 , Bettina E. Hansen 1 ; 1 Gastroenterology & Hepatology, 

Erasmus MC, University Medical Center, Rotterdam, Netherlands; 

2 Academic Medical Center, Amsterdam, Netherlands; 3 Toronto 

Western & General Hospital, Toronto, ON, Canada; 4 University 

of Padua, Padua, Italy; 5 University of Birmingham, Birmingham, 

United Kingdom; 6 Hôpital Saint-Antoine, APHP, Paris, France; 7 UT 

Southwestern Medical Center, Dallas, TX; 8 Humanitas Clinical and 

Research Center, Rozzano, Italy; 9 Università degli Studi di Milano, 

Milan, Italy; 10 , University of Barcelona, Barcelona, Spain; 11 University 

Hospitals Leuven, KULeuven, Leuven, Belgium; 12 The Royal 

Free Hospital, London, United Kingdom; 13 University of Alberta, 

Edmonton, AB, Canada; 14 Swedish Medical Center, Seattle, WA; 

15 Ehime University graduate School of Medicine, Ehim, Japan; 

16 Mayo Clinic, Rochester, MA; 17 Arizona State University, Phoenix, 

AZ 

Background and aim: Earlier reports have shown that patients 

with primary biliary cirrhosis (PBC) and low alkaline phosphatase 

(ALP) and bilirubin levels after 1 year of ursodeoxycholic 

acid (UDCA) treatment have favourable liver transplant-free survival 

. However, ALP and bilirubin may increase later during follow-up. 

The aim of the study was to identify patients within this 

group at risk of adverse outcome during follow-up. Methods: 

Patient data were obtained from the international Global PBC 

study group database, comprising long-term follow-up data 

of 4845 patients from 15 centers across North America and 

Europe. UDCA-treated patients with ALP ≤2.0xULN or bilirubin 

≤1.0xULN at 1 year follow-up were included in this analysis. 

Time-dependent Cox regressions analysis models was applied. 

Results: During a median follow-up of 7.11 years (IQR, 3.34- 

11.18) 350/2527 patients underwent liver transplantation or 

died. Patients who developed elevation of ALP (>2xULN) during 

follow-up had an increased risk of liver transplantation/death 

compared with those who remained ≤2.0xULN (HR 2.20, 95% 

CI 1.73-2.81, P value 1xULN) during follow-up were also at 

higher risk of reaching an endpoint (HR 5.51, IQR, 4.40-6.88, 

P value 2xULN, 25% had an 

event within 6.4 years; for elevated bilirubin levels this was 

1.5 years. Elevated ALP proofed to be an early predictor of 

abnormal bilirubin levels during follow-up; of patients with both 

normal bilirubin levels and ALP levels ≤2.0xULN 407 developed 

elevated bilirubin. Those patients who reached ALP levels 

>2.0xULN during follow-up were at higher risk of developing 

hyperbilirubinaemia (HR 3.79, 2.95-4.86, P value 2.0xULN) during 

follow-up is an early predictor of hyperbilirubinaemia, and an 

important early marker of a possible clinical event. 

Disclosures: 










Marlyn J. Mayo - Grant/Research Support: Intercept, Salix, NGM, Lumena, 

Gilead 

Albert Pares - Consulting: Lumena Pharmaceuticals, Intercept Pharmaceuticals, Inc 

Frederik Nevens - Consulting: MSD, CAF, Intercept, Gore, BMS, Abbvie, Novartis, 

MSD, Eumedica, Janssen, Promethera Biosciences; Grant/Research Support: 

Ferring, Roche, Astellas, Novartis, Janssen-Cilag, Abbvie 






Gilead 

Palak J. Trivedi - Grant/Research Support: Wellcome Trust 

The following authors have nothing to disclose: Willem J. Lammers, Annarosa 

Floreani, Gideon Hirschfield, Christophe Corpechot, Pietro Invernizzi, Pier Maria 

Battezzati, Douglas Thorburn, Angela C. Cheung, Teru Kumagi, Raoul Poupon, 

Ana Lleo, Llorenç Caballeria, Keith D. Lindor, Maren H. Harms, Bettina E. Hansen 

635 

Evaluation of the Posology of Obeticholic Acid (OCA) in 

Patients with PBC 

Richard Pencek, Karen Lutz, Tonya Marmon, Leigh MacConell; 

Intercept Pharmaceuticals, San Diego, CA 

Background: Obeticholic acid (OCA) is a potent and selective 

FXR agonist developed for treatment of primary biliary cirrhosis 

(PBC). 216 patients were enrolled in the Phase 3, double-blind, 

placebo-controlled trial and randomized to: Placebo (PBO), 10 

mg OCA (maximally efficacious dose in PBC), or 5 mg OCA 

titrating to 10 mg after Month (M) 6 based on tolerability and 

clinical response. Titration of OCA at M6 mitigated tolerability 

(pruritus, the most common AE) without compromising efficacy. 

While both 5 mg and 10 mg resulted in clinically meaningful 

and significant reductions in ALP compared to PBO, efficacy 

was greater at the 10 mg dose. This analysis evaluated the tolerability 

and efficacy Phase 3 data to support titration of OCA 

earlier than M6 after initiating OCA treatment. Methods: Time 

to first onset of treatment-emergent pruritus included the number 

of patients with pruritus (first onset), without pruritus (censored), 

and the minimum and maximum times in days. Kaplan-Meier 

estimates were plotted as a “survival curve” for each treatment 

group. Liver biochemistry measures (ALP, total bilirubin, GGT, 

ALT, and AST) were evaluated by LS mean change over time 

during the 12-month treatment period. Results: A Kaplan-Meier 

survival curve demonstrated that median time to first pruritus 

event was within the initial month of starting OCA. The median 

time to first onset of any pruritus event occurred within 2 weeks 

(OCA 10 mg), M1 (OCA 5 mg), and M2 (PBO). Mean BL 

ALP (U/L) was 327 (PBO) and 316 (OCA 10 mg). Clinically 

meaningful and significant improvements in ALP were achieved 

within 3M and maintained through the 12M period (Figure). 

Similar trends were observed in other liver biochemistry markers. 

Conclusions: The timing of onset of pruritus is early for both 

OCA doses. The majority of efficacy is observed within the 

first 3M of initiating OCA therapy. Thus, 3M after initiation of 

OCA is a reasonable timeframe to assess if a PBC patient may 

benefit from uptitration of OCA 5 mg to 10 mg.


around these tools particularly amongst non-specialists. Implementing 

a stratified approach to management requires these 

gaps to be addressed. 

Disclosures: 


Falk, Shire 




The following authors have nothing to disclose: Margaret Corrigan, Luke Vale, 

Diarmuid Coughlan 

Disclosures: 





The following authors have nothing to disclose: Karen Lutz, Leigh MacConell 

636 

Clinician confidence in stratifying risk in primary biliary 

cirrhosis – a UK-PBC survey 

Margaret Corrigan 1 , Luke Vale 2 , Diarmuid Coughlan 2 , David 

Jones 3 , Gideon Hirschfield 1 ; 1 NIHR Liver Biomedical Research 

Unit, University of Birmingham, Birmingham, United Kingdom; 

2 Institute of Health and Society, Newcastle University, Newcastle, 

United Kingdom; 3 Institute of Cellular Medicine, Newcastle Uiniversity, 

Newcastle, United Kingdom 

Background: Primary biliary cirrhosis (PBC) has only one present 

licensed therapy, ursodeoxycholic acid (UDCA). Future 

therapies will be offered to patients at high risk of disease progression. 

Assessment of biochemical response to UDCA allows 

identification of high risk patients. Aim: To survey current 

understanding of UDCA response criteria in the UK. Methods: 

A survey of current clinical practice was created by UK-PBC 

and distributed to clinicians via the British Society of Gastroenterology 

(BSG) and British Association for the Study of the 

Liver (BASL) mailing lists and newsletters. 206 responses were 

received from 1900 invites. Questions covered diagnosis and 

management of the condition with four questions specifically 

covering UDCA response assessment. Results: Respondents 

came from a variety of clinical backgrounds - consultant hepatologists 

in tertiary centres – 14 (7%), consultant hepatologists 

in non tertiary centres – 32 (15.5%), consultant gastroenterologists 

– 75 (36.4%), trainees – 78 (37.9%), others including specialist 

nurses - 7 (3.4%). Whilst 90% of respondents reported 

routine use of UDCA in clinical practice, only 20% reported 

always assessing UDCA response once patients have been 

on treatment for 12 months whilst 50% never assess response. 

Looking at rates of assessment of UDCA response between the 

specialist groups: 64% of gastroenterologists and 47% trainees 

never assess response compared to 25% of non-tertiary 

hepatologists and 14% of tertiary hepatologists. The number of 

patients seen appeared to affect UDCA response assessment: 

64% of those who saw fewer than 10 patients/year never 

assess response compared to 10% of those who saw more than 

50 patients/year. 40% of respondents reported themselves as 

‘not all confident’ in assessing response with 58% stating they 

were unaware that criteria were available and 27% unsure 

which criteria were best to use. Conclusion: The majority of 

patients with PBC are cared for in non-tertiary centres and most 

are managed by non-specialist clinicians. The application of 

emerging therapies for patients with PBC requires appropriate 

use of risk stratification tools in routine clinical practice. 

Our results demonstrates gaps in knowledge and confidence 

637 

The IgG/IgG4 mRNA ratio by quantitative PCR accurately 

diagnoses IgG4-related disease and predicts 

treatment response 

Lowiek M. Hubers 1 , Marieke E. Doorenspleet 2,3 , Emma L. Culver 

4,5 , Lucas Maillette de Buy Wenniger 1 , Paul L. Klarenbeek 2,3 , 

Roger W. Chapman 4,5 , Stan F. van de Graaf 1 , Joanne Verheij 

6 , Thomas van Gulik 7 , Frank Baas 3 , Eleanor Barnes 4,5 , Niek 

de Vries 2 , Ulrich Beuers 1 ; 1 Gastroenterology & Hepatology and 

Tytgat Institute of Liver and Intestinal Research, Academic Medical 

Center, Amsterdam, Netherlands; 2 Clinical Immunology & 

Rheumatology and Amsterdam Rheumatology and Immunology 

Center, Academic Medical Center, Amsterdam, Netherlands; 

3 Genome Analysis, Academic Medical Center, Amsterdam, Netherlands; 

4 Translational Gastroenterology Unit, John Radcliffe Hospital, 

Oxford, United Kingdom; 5 NDM Oxford University, Peter 

Medawar, Oxford University, Oxford, United Kingdom; 6 Pathology, 

Academic Medical Center, Amsterdam, Netherlands; 7 Surgery, 

Academic Medical Center, Amsterdam, Netherlands 

Introduction: IgG4-associated cholangitis (IAC) and autoimmune 

pancreatitis (AIP) are major manifestations of IgG4-related 

disease (IgG4-RD). Misdiagnosis and inadequate 

treatment are common since IAC and AIP mimic other inflammatory 

and malignant pancreatobiliary diseases, and accurate 

diagnostic biomarkers are lacking. Moreover, since relapse 

after tapering of immunosuppressive therapy occurs in 50% 

of patients, there is a need for biomarkers monitoring disease 

activity. Recently, using Next-Generation Sequencing, 

we observed that dominant IgG4+ B-cell receptor clones in 

peripheral blood distinguish patients with active IAC/AIP 

from primary sclerosing cholangitis (PSC) and pancreatobiliary 

malignancies (CA) [Hepatology 2013;57:2340]. Here, 

we report on a simple quantitative PCR (qPCR) protocol for 

diagnosing IAC/AIP and monitoring disease activity. Patients 

and Methods: 15 patients with IAC and/or AIP according to 

HISORt criteria, 7 patients with PSC and 8 with CA formed the 

test cohort. Intra- and extramural replication cohorts consisted 

of 16 IAC/AIP, 5 PSC and 13 CA patients (Dutch cohort), and 

8 IAC/AIP and 8 PSC patients (British cohort). In 20 Dutch 

IAC/AIP patients, follow-up samples after 4 and 8 weeks of 

corticosteroid therapy were available. RNA was isolated and 

the constant region of the B-cell receptor was amplified using a 

generic forward IgG primer together with either a generic IgG 

or a IgG4-specific reverse primer. The ratio total IgG/IgG4 

mRNA was calculated and expressed as ΔC T 

. Results: ΔC T 

as 

measure of IgG/IgG4 mRNA expression in peripheral blood 

of the test cohort was 2.8±1.1 (mean+SD) in IAC/AIP patients, 

compared to 6.8±1.6 in PSC and 7.6±1.4 in CA (Figure 1A, 

p


sion: IgG4-RD of the biliary tree and pancreas can be accurately 

distinguished from PSC or pancreatobiliary malignancies 

by ΔC T 

based on an affordable qPCR test. ΔC T 

can also be 

used as a marker for treatment response and disease activity 

in IAC and AIP. 

Figure 1. 

Disclosures: 




The following authors have nothing to disclose: Lowiek M. Hubers, Marieke E. 

Doorenspleet, Emma L. Culver, Lucas Maillette de Buy Wenniger, Paul L. Klarenbeek, 

Roger W. Chapman, Stan F. van de Graaf, Joanne Verheij, Thomas van 

Gulik, Frank Baas, Eleanor Barnes, Niek de Vries 

638 

Emperipolesis mediated by CD8+ T cells correlated with 

biliary epithelia cell injury in primary biliary cirrhosis 

Suxian Zhao, Rong-qi Wang, Yuguo Zhang, Huijuan Du, Yuemin 

Nan; Third Hospital of Hebei Medical University, Shijiazhuang, 

China 

Introduction: Primary biliary cirrhosis (PBC) is thought to be an 

autoimmune disease characterized by chronic destruction of 

bile ducts. A major unanswered question regarding the pathogenesis 

of PBC is the precise mechanisms of small duct injury 

and destruction. Emperipolesis is one of cell-in-cell structures 

that have been observed in chronic viral hepatitis. However, 

it remains unknown whether emperipolesis is involved in biliary 

epithelia cell injury and it diagnostic value in PBC. Aims: 

To clarify the pathogenesis and histological characteristics of 

emperipolesis in the liver injury of PBC. Methods:Sixty PBC 

patients, diagnosed by liver biopsy, were divided into two 

groups, early PBC (stages I and IIn=40) and late PBC (stages III 

and IV, n=20). The degrees of hepatic inflammation and fibrosis 

were assessed with Scheuer Scoring System. Emperipolesis 

was observed in liver sections stained with hematoxylin-eosin. 

The expression of CK19 (cholangiocyte), CD8 (T cell), CD20 (B 

cell), CD56 (NK cell), CD68 (macrophage cell) and MPO (neutrophil) 

was evaluated by mmunofluorescence double labeling 

combined with confocal microscopic observation. The apopotosis 

of BECs was detected by TUNEL assay under laser confocal 

microscope. Results: Emperipolesis was observed in 73.3% 

of patients with PBC, and BECs were predominantly host cells. 

In PBC patients with emperipolesis, the number of emperipolesis 

structures correlated with lymphoid follicles (r=0.4394, 

P


The following authors have nothing to disclose: Guo-Xiang Yang, Takashi Tomiyama, 

Ying Sun, Weici Zhang, Patrick S. Leung, Xiao-Song He, Sandeep S. 

Dhaliwal, M. Eric Gershwin 

640 

Natural killer cells regulate T cell immunity in primary 

biliary cirrhosis 

Shinji Shimoda 1 , Minoru Nakamura 2 , M. Eric Gershwin 3 ; 1 Kyushu 

University, Fukuoka, Japan; 2 Nagasaki University, Nagasaki, 

Japan; 3 UC Davis, Davis, CA 

BACKGROUND: Dissection of the multi-lineage response that 

leads to biliary epithelial destruction is essential for future therapeutic 

efforts. Indeed, a variety of work has already reflected 

that there are autoreactive T and B cells that do target biliary 

epithelial cells (BEC), but there is also an emerging set of data 

that suggests innate immunity is critical not only for the initiation, 

but also for a variety of interactions that alter the natural 

history of cholangitis. Our previous work has demonstrated that 

biliary cell cytotoxity is dependent on the initiation of innate 

responses followed by chronic adaptive as well as bystander 

mechanisms which are the interactions between natural killer 

(NK) cells and BEC. OBJECTIVE: We have taken advantage 

of our ability to isolate NK, BEC and endothelial cells from 

explanted liver samples, and studied the interactions between 

NK cells and BEC and focused on the mechanisms that activated 

autoreactive T cells in the presence or absence of antigen 

presenting cells (APC) and their dependence on IFN-g as 

well as expression of BEC HLA class I and class II molecules. 

Explanted liver and spleen from a total of 7 patients included 

2 with PBC and 5 with hepatitis C virus infection were used. 

There were no significant differences in the results derived from 

the liver mononuclear cells or the BEC population from either 

PBC or hepatitis C and ultimately all data were combined. This 

is consistent with our thesis that the BEC is an innocent victim 

as well as the established data that PBC reoccurs following 

transplantation. RESULTS: At a high NK/BEC ratio, NK cells 

attack autologous BEC in either a PBC-specific nor auto antigen-specific 

manner, and as glutathiolation does not occur in 

apoptotic BEC, autoantigen PDC-E2 that exists in the mitochondrial 

membrane is not degenerated by caspases in PBC, PDC- 

E2 from lysed BEC activate autoreactive CD4 positive T cells 

in the presence of APC. In contrast, at a low NK/BEC ratio, 

BEC were not lysed whereas IFN-g production was induced 

from NK cells, which in turn induced expression of HLA class 

I and II molecules on BEC and protected them from lysis upon 

exposure to autoreactive NK cells. In addition, IFN-g secretion 

from NK cells after exposure to autologous BEC enabled autoreactive 

CD4 positive T cells to lyse BEC in the presence of 

PDC-E2 antigen. CONCLUSION: Our data suggest that NK cell 

mediated innate immune responses are critical for the breach 

of tolerance at the initial stage of PBC, but also maintained the 

cytotoxic effect of autoantigen-specific T cells, which are critical 

for disease progression. 

Disclosures: 

The following authors have nothing to disclose: Shinji Shimoda, Minoru 

Nakamura, M. Eric Gershwin 

641 

Combination Anti-Retroviral Therapy Provides Reduction 

in Human Betaretrovirus Load and Durable Biochemical 

Responses in Patients with Primary Biliary Cirrhosis 

Ellina Lytvyak, Aldo J. Montano-Loza, Lynora Saxinger, Andrew 

Mason; University of Alberta, Edmonton, AB, Canada 

Purpose/Background: A human betaretrovirus has been characterized 

in primary biliary cirrhosis (PBC). Reverse transcriptase 

inhibitors lamivudine and zidovudine have shown limited 

efficacy. Whereas combination lopinavir/ritonavir (LPRr) and 

tenofovir/emticitabine (TDF/FTC) has been reported to be 

efficacious in normalizing liver tests (Lancet 2011). Methods: 

PBC patients unresponsive to UDCA were randomized into 

a crossover study with daily TDF/FTC 300/200mg and LPRr 

800/200mg versus placebo for 6 months; followed by an 

open label study for a total of 24 months. HBRV DNA was 

assessed in PBMC using digital droplet PCR. Results: (A) In the 

6 month RCT, a significant reduction in Alk Phos was observed 

in patients on TDF/FTC and LPRr (p


642 

Long term impact of fibrates in primary sclerosing cholangitis 

with incomplete biochemical response to ursodeoxycholic 

acid : the French-Spanish experience 

Sara Lemoinne 2,1 , Christophe Corpechot 2,1 , Astrid Donald D. 

Kemgang Fankem 2,1 , Farid Gaouar 2 , Raoul Poupon 2,1 , Chantal 

Housset 2,1 , Albert Pares 3 , Olivier Chazouillères 2,1 ; 1 INSERM, 

UMRS_938, F-75012 Paris, France UPMC Univ Paris 06, F-75012 

Paris, France, Paris, France; 2 Service d’Hépatologie, Centre de 

Référence des Maladies Inflammatoires des Voies Biliaires, Hôpital 

Saint-Antoine, Assistance-Publique Hôpitaux de Paris, Paris, 

France; 3 Liver Unit, Hospital Clinic, University of Barcelona, 

IDIBAPS, CIBERehd, Barcelona, Spain 

Background : In patients with primary sclerosing cholangitis 

(PSC), ursodeoxycholic acid (UDCA) improves serum livers tests 

and some surrogate markers of prognosis. In the absence of 

medical treatment with proven efficacy, UDCA is widely used 

in European PSC patients. However, newer therapies are obviously 

needed. Fibrates, PPAR agonists that exert anti-inflammatory 

properties in several experimental models of autoimmunity, 

seem to have a beneficial effect on liver biochemistries in primary 

biliary cirrhosis. In 2014, we reported improvement of 

liver tests under fibrates in 15 French PSC patients with an 

incomplete biochemical response to UDCA. Aim : To confirm 

the safety and efficacy of fibrates in an extended PSC population 

including patients from another center. Methods : This 

retrospective study included patients with PSC treated with 

fibrates (fenofibrate 200mg/d or bezafibrate 400mg/d) for 

at least 6 months in addition to UDCA, after an incomplete 

biochemical response (ALP>1ULN) to UDCA (15-20mg/kg/d) 

for at least 1 year. Patients with associated liver diseases, especially 

auto-immune hepatitis were not included. Changes in biochemical 

parameters have been assessed using a linear mixed 

model and Wilcoxon tests. Results : 26 patients were included 

(19 from Paris and 7 from Barcelona) : 17 males, median age 

49.7 years, 16 with inflammatory bowel disease, median liver 

stiffness 10.6 kPa (corresponding to fibrosis ≥ F3). Median 

duration of treatment with fibrates was 36.8 months (6.7- 

61.3). Under treatment with fibrates, ALP and ALT decreased 

significantly (p= 0.04 and 0.01 respectively) and this decrease 

was maintained at 48 months of treatment. GGT tended to 

decrease (p=0.10). AST, total bilirubin and albumin remained 

unchanged. No serious adverse event related to fibrates 

occured. In 54% of patients, treatment by fibrates was stopped 

because of various reasons : development of biliary stones, 

gastro-intestinal bleeding, decision by cardiologist, liver tests 

worsening. Excluding this latter group, interruption of fibrates 

was followed by a significant increase of ALP. During follow-up 

of a median duration of 4.1 years, 2 cholangiocarcinoma, 5 

liver transplantations and 1 death were recorded. Interestingly, 

despite a biochemical improvement, liver stiffness assessed by 

transient elastography increased significantly under fibrates. 

Conclusion : Addition of fibrates induces a significant biochemical 

improvement in PSC patients with incomplete biochemical 

response to UDCA but seems to be insufficient to control the 

progression of advanced PSC. This raises again the critical 

issue of surrogate markers of prognosis in PSC clinical trials. 

Disclosures: 

Albert Pares - Consulting: Lumena Pharmaceuticals, Intercept Pharmaceuticals, Inc 


The following authors have nothing to disclose: Sara Lemoinne, Christophe 

Corpechot, Astrid Donald D. Kemgang Fankem, Farid Gaouar, Raoul Poupon, 

Chantal Housset 

643 

Predictors of recurrence and subsequent outcomes for 

patients admitted with a sentinel vs recurrent episode of 

acute cholangitis 

Amanda M. Lynn 1 , James H. Tabibian 3 , Todd H. Baron 2 ; 1 Internal 

Medicine, Mayo Clinic, Rochester, MN; 2 Gastroenterology and 

Hepatology, University of North Carolina, Chapel Hill, NC; 3 Gastroenterology 

and Hepatology, Mayo Clinic, Rochester, MN 

Introduction: Acute cholangitis (AC) is well-recognized as a 

morbid and potentially lethal condition. Previous studies have 

investigated predictors of disease severity and short-term outcomes; 

however, data on long-term outcomes such as risk of 

recurrence and subsequent out-of-hospital mortality are lacking. 

We investigated potential differences in short and longterm 

outcomes of patients admitted with a sentinel episode 

of AC compared to those with a history of AC. Methods: We 

retrospectively identified all patients admitted to Mayo Clinic 

hospitals in Rochester, MN for AC from 2009-2011. Patients 

were categorized into two groups based on the presence or 

absence of prior AC. Data on pertinent clinico-demographic 

variables and outcomes including hospital length-of-stay (LOS), 

AC recurrence, and 1-year mortality were abstracted using a 

standardized data collection form. Tests of significance were 

2-tailed, and p


644 

Long-Term Safety of OCA in Patients with PBC 

Yvette Peters, Roya Hooshmand-Rad, Richard Pencek, Janet 

Owens-Grillo, Tonya Marmon, Leigh MacConell, David Shapiro; 

Intercept Pharmaceuticals, San Diego, CA 

Background: Obeticholic Acid (OCA) is a potent and selective 

FXR agonist developed for treatment of primary biliary cirrhosis 

(PBC). 216 patients with PBC were treated in a randomized, 

doubleblind (DB), placebo (PBO)controlled Phase 3 clinical 

study to evaluate the efficacy and safety of OCA. 198 patients 

completed the DB phase of the study and 193 enrolled in a 

longterm extension (LTSE) phase. Exposure to OCA during the 

DB phase resulted in statistically significant liver biochemistry 

improvements and was safe. Methods: All patients enrolled in 

the LTSE first met the inclusion criteria for the DB study, which 

included PBC diagnosis, ALP ≥1.67x ULN and/or total bilirubin 

>ULN to


646 

Significant non–adherence to DAA HCV therapy in 

decompensated patients - a tertiary hepatology centre 

assessment 

Aisling B. Considine 1,2 , Suman Verma 3 , Kath Oakes 3 , Kate E. 

Childs 3 , Sarah Knighton 1,2 , Andrew Ayers 3 , Abid Suddle 3 , Kosh 

Agarwal 3 ; 1 Pharmacy, Kings College Hospital, London, United 

Kingdom; 2 Kings College London, Institute of Pharmaceutical Sciences, 

London, United Kingdom; 3 Kings College Hospital, Institute 

of Liver Studies, London, United Kingdom 

Background: The advent of directly acting anti-virals (DAA) has 

led to simplified HCV regimens with increased efficacy rates 

and better tolerability. The high adherence seen in clinical 

trials may not translate to real life populations. Optimal adherence 

is critical to protect patients from treatment failure and 

resistance. Aim: To identify potential factors which may contribute 

to sub-optimal adherence in a population with decompensated 

cirrhosis. Methods: HCV patients eligible for access 

to a 12 week treatment regimen under a UK NHSE mandated 

access scheme were consented to complete a baseline questionnaire 

capturing data on sociodemographics, clinical status 

and perceptions of illness.Adherence assessments at treatment 

weeks 4, 8 and 12 with pill counts and a Morisky Medication 

Adherence Scale were undertaken. Suboptimal adherence 

(SA) was defined as any report or pill count which indicated 

a delayed (>2hrs) or missed dose of anti-viral. Descriptive 

statistic and multivariate regression methods were utilised for 

analysis. Results: 80% (n=47) of the cohort had reached SVR 

12 and are included in the adherence analysis.85% received 

sofosbuvir/ledipasvir and 15% sofosbuvir with daclatasvir. All 

patients received ribavirin and had been attending specialist 

hepatology services for >12 months.43% (n=20) of patients 

demonstrated SA during their treatment. Multivariate analysis 

demonstrated that any non-attendance during treatment 

and a ‘limited support network’ predicted higher rates of SA 

(p3 medications 

(vs 74% in adherent group). The average MELD score was 

15 in the SA group (vs 13 in adherent group). 80% of SA 

was associated with symptoms secondary to disease stage not 

reflected by the MELD. SA secondary to side effects of therapy 

was not identified. 85% of the SA group achieved SVR12.Failure 

to achieve SVR was seen in those with SA adherence at all 

assessment points (weeks 4, 8 and 12). Conclusions: Our initial 

results demonstrate that despite an intensive adherence focused 

multi-disciplinary approach, SA occurs in decompensated HCV 

cirrhotic patients. Acceptable SVR rates were achieved in this 

population. Further research is warranted to develop strategies 

which maximise adherence in all HCV populations. 

Disclosures: 




The following authors have nothing to disclose: Aisling B. Considine, Suman 

Verma, Kath Oakes, Kate E. Childs, Sarah Knighton, Andrew Ayers, Abid Suddle 

647 

Hepatology nursing in Canada: Current practices and 

future challenges 

Colina Yim 1 , Cheryl Dale 2 , Geri Hirsch 3 , Jo-Ann E. Ford 4 , Carolyn 

Klassen 4 ; 1 University Health Network, Toronto, ON, Canada; 

2 London Health Sciences Centre, London, ON, Canada; 3 Halifax 

Capital District Health Authority, Halifax, NS, Canada; 4 Vancouver 

General Hospital, Vancouver, BC, Canada 

Background: Hepatitis C treatment landscape is changing. 

The rapid therapeutic advancements may impact the roles and 

responsibilities of some of the hepatology nurses whose practice 

focused solely on hepatitis C. This study aims to describe 

the current practices of hepatology nurses in Canada and 

to gain insights into nurses’ perceptions and concerns about 

their future roles and values. Methods: An 18-item survey was 

designed. Survey was sent in November 2014 to all active 

members of the Canadian Association of Hepatology Nurses 

(CAHN) via email for their voluntary participation. A second 

email was sent 8 weeks later as a reminder to complete survey. 

Demographics collected include age, length of practice, 

practice settings and focuses. CAHN Members were asked to 

rate their levels of concerns in regards to job security, sustainability 

of current positions, change of role and the future of 

hepatology nursing. Results: Of a total of 136 active CAHN 

members, 94 (69%) completed the survey. Eighty-one percent 

were employed full time, 78% age > 40 years, 64% have at 

least 6 years or more work experience in hepatology. Hepatitis 

C treatment was the sole practice focus in 52% while 38% of 

them managed more than 50 hepatitis C patients on treatment 

in last 6 months. A majority of CAHN members (78%) work 

in academic institutions and community settings. Salary support 

varies with 35% being paid from industry grants and the 

remainder either by government (health authority, provinces, 

academic centres and community agencies) or by research 

grants. Up to 85% expressed satisfaction with their current 

positions and the support they received from their physician 

partners but at least 60% are concerned about their job security, 

job description change and current position sustainability. 

A lack of funding for hepatology nurse positions remain the 

major challenge, 69% of members predict a positive hepatology 

nursing future. Conclusions: Hepatology nurses in Canada 

are experienced professionals in the field. A majority have a 

focused practice on hepatitis C treatment management. We 

are satisfied with our current roles and responsibilities and 

feel most supported by our physician colleagues. Hepatology 

nurses acknowledge challenges exist for role sustainability but 

remain optimistic about our values in future. 

Disclosures: 

Colina Yim - Advisory Committees or Review Panels: Janssen; Consulting: Gilead; 

Speaking and Teaching: Abbvie 

Jo-Ann E. Ford - Advisory Committees or Review Panels: Gilead, Roche, Janssen, 

Merck Canada, Vertex; Speaking and Teaching: Gilead, Roche, Janssen, Merck 

Canada, Vertex 

The following authors have nothing to disclose: Cheryl Dale, Geri Hirsch, Carolyn 

Klassen 

648 

Pharmacists mitigate effect of simeprevir on blood pressure 

in patients with hepatitis C infection receiving concurrent 

calcium channel blocker therapy 

Heather Johnson 1,2 , Emily Graham 2 , Michael A. Dunn 1 , Kapil B. 

Chopra 1 ; 1 University of Pittsburgh, PIttsburgh, PA; 2 UPMC Presbyterian, 


Simeprevir (SMV) an NS3/4A inhibitor for the treatment of 

Hepatitis C infection (HCV), has a number of drug-drug inter-


actions mediated by CYP3A4 or p-glycoprotein. Current product 

labeling indicates increased clinical monitoring of patients 

receiving SMV with oral calcium channel blockers (CCB), but 

no specific guidance is available. Pharmacists complete a 

medication review for all patients receiving therapy for HCV 

to identify potential drug-drug interactions and opportunities 

to mitigate interactions. We examined blood pressure control 

in patients on SMV and simultaneous CCB, regardless of 

mitigating interventions, between December 2014 and June 

2015 to determine if concurrent treatment with SMV and a 

CCB resulted in hypotension leading to adjustments in antihypertensive 

regimens or if hypertension resulted in patients who 

had preemptive CCB dose reductions before SMV initiation. 

Twenty-three patients were included. Of those, 65.2% were 

male and 95.6% were Caucasian. Patients were receiving an 

average of 5.2mg amlodipine equivalents at initiation of SMV. 

Doses of CCB were preemptively reduced in 9 (39%) patients 

prior to SMV initiation. Prior to preemptive dose reduction, the 

mean amlodipine equivalent in patients was 8.2 mg . All 9 

patients who received preemptive dose reduction of their CCB 

were concurrently taking other antihypertensive medications. 

Most were receiving one additional antihypertensive (7) with 1 

each receiving 2 and 3 other medications. In patients without 

CCB dose reduction, 6 were on one and 2 were on 2 other 

antihypertensive medications. Only one patient without preemptive 

CCB dose reduction discontinued CCB during SMV 

treatment. Two patients discontinued other antihypertensive 

therapies due to hypotension during concurrent treatment with 

SMV and a CCB. The average mean arterial pressure (MAP) in 

all patients prior to initiating SMV was 95 mmHg. The average 

MAP in all patients mid-treatment with SMV and after discontinuation 

was 93 and 90 mmHg, respectively. In patients who 

had preemptive dose reductions of their CCB, the MAP at baseline, 

mid-treatment, and after discontinuation were 94.3, 93.2, 

and 90.6 mmHg, respectively. Preemptive CCB dose reduction 

is a safe strategy in patients receiving SMV and did not lead to 

uncontrolled hypertension. Most patients receiving a CCB with 

SMV did not experience hypotension during therapy, however, 

further dose reduction may be warranted in select patients. 

Disclosures: 

The following authors have nothing to disclose: Heather Johnson, Emily Graham, 

Michael A. Dunn, Kapil B. Chopra 

649 

FGF21 facilitates normal liver regeneration and restores 

impaired liver regeneration in steatotic liver 

Hui-Xin Liu, Ying Hu, Yu-Jui Yvonne Wan; Medical Pathology and 

Laboratory Medicine, UC DAVIS, Sacramento, CA 

Fibroblast growth factor 21 (FGF21) is a master metabolic 

regulator that has a remarkable ability to reverse obesity, 

repair injured tissue, and prolong lifespan. The current study 

examined the effect of FGF21 in regulating 2/3 partial hepatectomy 

(PHx)-induced liver regeneration in normal diet and 

Western diet-fed mice. In vitro, adenovirus expressing FGF21 

(Ad-FGF21) was used to overexpress FGF21 in mouse primary 

hepatocytes. In the absence of serum, exogenous FGF21 

increased hepatocyte viability by 160% indicating a mitogenic 

effect. In mice, PHx resulted in transient induction of FGF21 

and activation of ERK1/2, which was associated with induction 

of cell cycle genes 1 Day after liver resection. Forced 

expression of FGF21 through tail vein injection with Ad-FGF21 

accelerated hepatocyte proliferation that was accompanied 

by earlier activation of ERK1/2 and induction of cell cycle 

genes compared to mice injected with control Ad-CMV. Mild 

steatosis, which is considered as an energy source for hepatocyte 

proliferation, was absent in Ad-FGF21-injected mice 

1-1.5 Days after PHx. Conversely, FGF21 knockdown using 

siRNA (siFGF21) increased the severity of steatosis, delayed 

the induction of cell cycle genes, and decreased the number 

of Ki67-positive cells. These findings suggested a close 

relationship between FGF21-regulated lipid metabolism and 

hepatocyte proliferation. Consistently, liver regeneration was 

delayed in steatotic livers induced by Western diet feeding. 

The impaired liver regeneration, accompanied by decreased 

Ki67-positive cells, reduced cell cycle gene expression, and 

steatosis, was further exacerbated by FGF21 knockdown. 

However, forced expression of FGF21 in Western diet-induced 

fatty livers rescued impaired liver regeneration as indicated 

by normalized cell cycle gene and lipid metabolizing gene 

expression patterns. Conclusion: FGF21 plays a pivotal role in 

regulating metabolism and liver regeneration. Forced expression 

of FGF21 facilitates normal regeneration and restores the 

regeneration program in Western diet-induced fatty liver. 

Disclosures: 

The following authors have nothing to disclose: Hui-Xin Liu, Ying Hu, Yu-Jui 

Yvonne Wan 

650 

HNF4α reprogramming during initiation and termination 

of liver regeneration 

Ian Huck, Michael W. Manley, Jr., Udayan Apte; Pharmacology, 

Toxicology and Therapeutics, University of Kansas Medical Center, 

Kansas City, KS 

Hepatocyte nuclear factor 4 alpha (HNF4α) is an orphan 

nuclear receptor essential for hepatocyte differentiation and 

function. Recent studies have indicated that HNF4α also 

inhibits hepatocyte proliferation and may be tumor suppressor. 

However, the role of HNF4α in liver regeneration is not 

known. We investigated changes in HNF4α expression and 

function in liver regeneration after partial hepatectomy (PH) 

using three-month-old male C57BL/6 mice. Real Time PCRbased 

analysis revealed presence of mRNA for both P1 and P2 

isoforms after PH but only the P1 (adult) form of HNF4α protein 

was detected in the liver by Western blot. A rapid decline in 

total and nuclear levels of HNF4α was observed within 1 hr 

after PH, which further decreased at 6 hr post-PH. Whereas, 

HNF4α protein increased at 12 hr post PH as compared to 6 

hr, it remained significantly lower than 0 hr and continued to 

be lower till 5 days after PH, at which time it returned to control 

(0 hr) levels. A coinciding decline in HNF4α function as measured 

by 44 select targets genes was observed within 12 hr 

after PH, which recovered substantially by 5 days post PH. We 

further studied the role of HNF4α is liver regeneration after PH 

using conditional hepatocyte specific HNF4α knockout mice 

(HNF4α-KO). Cell proliferation measured by PCNA analysis 

started 24 hr earlier in the HNF4a-KO mice as compared to 

WT and continued beyond 7 days after PH. HNF4α deletion 

resulted in an accelerated liver regeneration and induced a 

defect in termination of regeneration after PH. In conclusion, 

these data indicate that HNF4α undergoes significant reprogramming 

during liver regeneration after PH and is involved in 

both initiation and termination of liver regeneration. 

Disclosures: 

The following authors have nothing to disclose: Ian Huck, Michael W. Manley, 

Jr., Udayan Apte


651 

Skeletal muscle mitochondrial fragmentation results in 

functional abnormalities during hyperammonemia of 

cirrhosis 

Gangarao Davuluri 2 , Avinash Kumar 2 , Samjhana Thapaliya 2 , 

Allawy Allawy 2,5 , Dharmvir Singh 2 , Julie H. Rennison 2 , Rafaella 

Nascimento e Silva 2 , David R. Van Wagoner 3 , Sathyamangla V. 

Naga Prasad 3 , Xin Qi 4 , Hoppel Charles 4 , Takhar Kasumov 2 , Srinivasan 

Dasarathy 1 ; 1 Department Of Gastroenterology and Hepatology, 

Cleveland Clinic, Cleveland, OH; 2 Pathobiology, Cleveland 

Clinic, Cleveland, OH; 3 Molecular Cardiology, Cleveland Clinic, 

Cleveland, OH; 4 Pharmacology and Medicine, Case Western 

Reserve University, Cleveland, OH; 5 Department Of Internal Medicine, 

Cleveland Clinic, Cleveland, OH 

Background. Hyperammonemia is a consistent abnormality 

in cirrhosis and contributes to sarcopenia. Skeletal muscle is 

a major alternate organ for ammonia disposal by the initial 

reaction of conversion to glutamate in the mitochondria. We 

have observed skeletal muscle mitochondrial functional abnormalities 

in cirrhosis and hyperammonemia. It is not known if 

the functional abnormalities are due to structural alterations 

in the skeletal muscle mitochondria. Methods. Mitochondrial 

mass was measured by immunoblots for mitochondrial proteins 

citrate synthase, voltage dependent anion channels and cyclooxygenase 

IV during hyperammonemia in C2C12 myotubes. 

Additional methods to quantify mitochondrial mass included 

electron microscopy and immunofluorescence images using 

mitochondrial dye, MitoTracker Red. Mitochondrial fragmentation 

was quantified using electron microscopy and immunofluorescence 

images with Mitotracker red. Membrane polarization 

generated by proton pumping by components of the electron 

transport chain (ETC) was quantified using the dye TMRE and 

uncoupler, FCCP was used as a positive control. The mechanism 

of altered mitochondrial dynamics during hyperammonemia 

was determined by quantifying genes regulating mitochondrial 

fission and fusion by real time PCR and immunoblots of DRP1 

protein expression and polymerization polymerization. Finally, 

siRNA to knockdown of DRP1 and P110 peptide to block 

DRP1 function in myotubes were used to show that ammonia 

promoted DRP1 polymerization that resulted in mitochondrial 

fragmentation and dysfunction. Results. Despite unaltered mitochondrial 

mass with hyperammonemia, increased mitochondrial 

fragmentation was observed on both immunofluorescence 

and electron microscopy. These changes were accompanied 

by consistent alterations in expression of genes regulating mitochondrial 

dynamics, Fis 1, OPA1, MFN1, and MFN2. We 

also observed increased polymerization of dynamin related 

protein-1 (DRP1) during hyperammonemia in the myotubes. 

Genetic and pharmacologic inhibition of DRP1 in restored 

ATP content and decreased reactive oxygen species in myotubes 

regardless of hyperammonemia. These changes were 

accompanied by loss of membrane potential during hyperammonemia. 

Conclusions. Increased mitochondrial fragmentation 

was observed with unaltered mitochondrial mass and was due 

to polymerization of dynamin related protein-1 (DRP1). Since 

blocking mitochondrial fragmentation reversed the low ATP 

and elevated ROS during hyperammonemia, our observations 

provide a novel approach to reverse skeletal muscle bioenergetic 

dysfunction and sarcopenia of cirrhosis by targeting 

mitochondrial fragmentation using anti DRP1 therapies. 

Disclosures: 

The following authors have nothing to disclose: Gangarao Davuluri, Avinash 

Kumar, Samjhana Thapaliya, Allawy Allawy, Dharmvir Singh, Julie H. Rennison, 

Rafaella Nascimento e Silva, David R. Van Wagoner, Sathyamangla V. Naga 

Prasad, Xin Qi, Hoppel Charles, Takhar Kasumov, Srinivasan Dasarathy 

652 

Thr505 RelA phosphorylation controls liver proliferative 

response and supresses NF-κB tumor-promoting activities 

Anna Moles 1 , Jacqueline Butterworth 2 , Jill E. Hunter 2 , Ana M. 

Sanchez 2 , Dina Tiniakos 1 , Derek Mann 1 , Fiona Oakley 1 , Neil 

D. Perkins 2 ; 1 Institute of Cellular Medicine, Newcastle University, 

Newcastle Upon Tyne, United Kingdom; 2 Institute for Cell and 

Molecular Biosciences, Newcastle University, Newcastle Upon 

Tyne, United Kingdom 

Background and aims: NF-κB is a family of transcription factors 

that are key regulators of the immune and inflammatory 

responses. NF-κB regulation is complex and occurs at many 

levels, including direct post-translational modifications (PTMs) 

of the subunits. Phosphorylation of RelA (p65) at Thr505 regulates 

proliferation, migration and autophagy in in vitro cellular 

systems; however its role and relevance in vivo still remains 

unclear. Thus, the aim of this study was to analyse the role 

of RelA Thr505 phosphorylation in vivo by mutating this site 

through creating a knock in mouse. Methods: Knock in mutant 

mice where generated where Thr505 was substituted by Alanine, 

thus preventing phosphorylation. Partial hepatectomy 

(PhX), acute CCl 4 

administration and 15 days DEN injection 

was performed in WT and knock in mice. Animals were culled 

at 36, 72 hours and 5 days for PhX; at 24, 48 and 72 hours 

for acute CCl 4 

and 30 weeks post-DEN. Paraffin staining for 

BrdU and PCNA were used as markers of proliferation and 

γH2AX for DNA damage. Western blot was used to determined 

p53 and p-JNK expression. Gene set enrichment analysis 

(GSEA) was performed in WT and RelA T505 livers after 36 

hours of PhX. Tumor assessment was performed by an expert 

pathologist. Results: Thr505 mutation of RelA resulted in an 

increased liver to body weight ratio after PhX at 36, 72 hours 

and 5 days. Hepatocyte proliferation was increased after both 

injuries, PhX and acute CCl 4, 

at all the time points in RelA 

T505 mice versus WT, as assessed by PCNA and/or BrdU 

staining. Furthermore, mitotic body counts were also higher 

in RelA T505 mice after both injuries. Of note, proliferation 

after PhX was sustained for longer in RelA T505 mice, pointing 

towards an inefficient resolution of the proliferative response. 

Increased DNA damage, as determined by γH2AX staining, 

was seen in both WT and RelA T505 after 36 hours of PhX. 

However, the DNA damage was significantly higher in RelA 

T505 mice in comparison with WT. Western blot analysis of 

these samples revealed increased expression of the p53 tumour 

suppressor and active JNK in RelA T505 mice. Finally, chronic 

DEN administration resulted in a significant increased tumor 

number, mostly HCC, in RelA T505 versus WT. Conclusions 

RelA Thr505 phosphorlyation provides an important and novel 

regulatory mechanism to control the liver proliferative response 

and is a critical pathway suppressing NF-κB tumour-promoting 

activities of RelA. 

Disclosures: 

The following authors have nothing to disclose: Anna Moles, Jacqueline Butterworth, 

Jill E. Hunter, Ana M. Sanchez, Dina Tiniakos, Derek Mann, Fiona 

Oakley, Neil D. Perkins


653 

Lysophosphatidic acid receptor 3 (LPAR3) is expressed 

by cancer progenitor cells in human HCC tissue at the 

tumor:non-tumor margin and regulates cell migration. 

Valentina Zuckerman 1 , Eugene Sokolov 1 , Jacob H. Swet 1 , Victor 

C. Showalter 1 , William A. Ahrens 2 , David A. Iannitti 1 , Iain H. 

McKillop 1 ; 1 Department of Surgery, Carolinas Medical Center, 

Charlotte, NC; 2 Department of Pathology, Carolinas Medical Center, 

Charlotte, NC 

Background The poor prognosis for hepatocellular carcinoma 

(HCC) patients can be attributed, in part, to late detection, 

rapid tumor expansion, and metastases. LPAR3 is detected 

at low levels in the liver/ hepatocytes, and previous studies 

report LPAR3 expression increases in HCC and diseased/cirrhotic 

non-tumor liver (NTL) from HCC patients compared to 

healthy normal liver (NL). A growing interest in stem cells in 

tumor biology, and the role of LPAR3 in other cancers, led us 

to hypothesize that changes in LPAR3 expression in HCC may 

occur in a subset of hepatic progenitor cells involved in tumor 

progression/expansion. Methods Sections from human HCC- 

NTL, and lysates from SKHep1 cells (liver cancer progenitor 

cells) and NL were analyzed for LPAR3, CD44 (mesenchymal 

stem cell marker), EpCAM (cancer progenitor cell marker), 

and Hepar1 (hepatocyte marker) expression-localization. The 

effect of LPA on cell function was determined in SKHep1 using 

2D and 3D cell migration analysis. Cell signaling (AKT/MEK 

activity) and migration following LPA (10μM) treatment was 

analyzed in conjunction with inhibition of Gi-protein dependent 

signaling (PI3K inhibitor (LY294002 [LY], 40μM) and MEK 

inhibitor (U0126, 5μM)). In parallel shRNA was used to inhibit 

endogenous LPAR3 expression. Results Extensive co-localization 

of LPAR3 with CD44 and EpCAM was detected in the 

HCC-NTL margin, but not the NTL or tumor mass, data mirrored 

in SKHep1 cells (LPAR3, CD44 and EpCAM detected). Conversely, 

Hepar1 was readily detected in NL and the HCC mass, 

but not SKHep1 cells, or cells in the HCC margin that stained 

positive for LPAR3. In vitro analysis demonstrated LPA-dependent 

PI3K and ERK activation in SKHep1 cells resulting in 

significant cell migration. Pretreatment with LY and U0126 

abrogated LPA-dependent PI3K and MEK activation. However, 

inhibition of PI3K (LY) failed to significantly alter LPA-dependent 

migration whereas ERK inhibition (U0126) abolished LPA-dependent 

migration. The effects of pharmacological inhibition 

were mirrored by inhibiting LPAR3 expression (shRNA), in 

which exogenous LPA failed to stimulate ERK activation and 

cell migration. Summary These data identify a unique subset 

of LPAR3 positive cancer progenitor cells at the tumor–NTL 

margin in HCC patients. Using an in vitro model of hepatic 

tumor progenitor (SKHep1) cells, with similar LPAR profiles to 

in vivo tissue, we report LPA regulates tumor cell migration via 

Gi-MEK-ERK dependent signaling. These data suggest targeting 

LPA-LPAR3 signaling within this progenitor cell subset in the 

liver may slow tumor expansion and/or metastasis and limit 

disease progression. 

Disclosures: 

David A. Iannitti - Consulting: Davol, Covidien, Ethicon Endosurgery, Davol, 

Covidien, Ethicon Endosurgery, Davol, Covidien, Ethicon Endosurgery, Davol, 

Covidien, Ethicon Endosurgery; Speaking and Teaching: Davol, Covidien, 

Ethicon Endosurgery, Davol, Covidien, Ethicon Endosurgery, Davol, Covidien, 

Ethicon Endosurgery, Davol, Covidien, Ethicon Endosurgery 

The following authors have nothing to disclose: Valentina Zuckerman, Eugene 

Sokolov, Jacob H. Swet, Victor C. Showalter, William A. Ahrens, Iain H. McKillop 

654 

Hepatitis B virus X protein promotes HCC metastasis 

through activation of CD44v6 splicing form switching by 

regulation of splicing factor ESRP1/ESRP2 

Ya-Ju Chang 1 , Li-Chen Wu 2 , Ja-an Annie Ho 1 ; 1 Department of 

Biochemical Science and Technology, National Taiwan University, 

Taipei, Taiwan; 2 Department of Applied Chemistry, National Chi 

Nan University, Puli, Taiwan 

Purpose Hepatitis B virus x (HBx) has been implicated in the 

development of hepatocellular carcinoma (HCC) through the 

promotion of malignant transformation, upregulation of the 

properties of liver cancer stem cells (CSC), and manipulation 

of cell cycle progression, but it is not clear how HBx promotes 

HCC metastasis. Among CSC markers, CD44 is associated 

with migration, proliferation, and invasion, especially CD44v6, 

which is critical for metastasis. Herein the effects of HBx on 

CD44v6-dependant HCC metastasis was investigated. Methods 

Stable tet-on inducible HBx overexpression cells were obtained 

by transfection of pLV-CMVTO-GFP-HBx plasmids to HepG2 

for studying HCC metastasis. The expression of CD44v6 was 

induced by doxycycline (Doxy.) and was determined by immunoblotting, 

quantitative real-time PCR (Q-PCR), and flow cytometry. 

The metastatic ability of HBx-induced HCC cell lines was 

investigated by real-time cell analysis. The effect of HBx/snail 

complex on the expression of CD44 splicing factor ESRP1 and 

ESRP2 was analyzed. The inhibitory effects of siESRP1 and 

siESRP2 were also studied. Xenograft tumor model was used for 

in vivo study for the investifation of the association of CD44v6 

in metastasis and the splicing factor on HBx expressed tumors. 

Results HBx significantly induced the expression of CD44v6 

as identified by immunoblotting, Q-PCR, and flow cytometry. 

Through interaction with snail, HBx promoted the expression 

of downstream signaling CD44-related splicing factor ESRP2, 

and restricted ESRP1. Inhibition of snail and ESRP2 but not 

ESRP1 markedly reduced the mobility of HBx-expressed cells. 

Addtionally, overexpressed N-cadherin induced by HBx but not 

E-cadherin was detected, indicating the undergoing of EMT. 

MMP2 and 9 were also evaluated for the potential of metastasis. 

HBx enhanced the expression of these two metalloproteinases. 

Futher metastasis was confirmed by enhanced RhoA-GTP, 

a ROCK family protein participating in cell migration/invasion, 

determined by IP/IB in HBx expressed HepG2, and cell metastatic 

ability as determined by real-time cell monitoring. Based 

on these findings, HBx appears to significantly promote HCC 

invasion through snail modulated ESRP1/ESRP2, which might 

be a driven force to enhance CD44v6-dependent metastatic 

ability of HCC. Conclusion HBx was able to promote metastatic 

ability in HCC cell line through epigenetic regulation. The metastatic 

modification of CD44 variant was through the ESPR1 

and ESRP2, that function as specific modulators in CD44 splicing 

to maneuver metastatic ability. Inhibition of CD44 splicing 

seems to be a potential approach to arrest the progression of 

HBV-related HCC metastasis. 

Disclosures: 

The following authors have nothing to disclose: Ya-Ju Chang, Li-Chen Wu, Ja-an 

Annie Ho


655 

Regulation of the Hepatic Drug-metabolizing Enzymes 

by Probiotics and Conventionalization in Germ-free 

Mice 

Felcy pavithra Selwyn samraj, Sunny Lihua Cheng, Curtis Klaassen, 

Julia Yue Cui; Environmental and Occupational Health Sciences, 

University of Washington, Seattle, WA 

The use of probiotics and antibiotics has raised concerns of 

potential adverse “drug-bacteria” interactions in humans. VSL3 

is a probiotic medication that delivers 8 live strains of bacteria 

to the host, and has been used to treat ulcerative colitis. The 

present study utilized conventional (CV) and germ-free (GF) 

mice to determine the effects of VSL3 and conventionalization 

of GF mice by exposing them to the CV housing environment 

on the expression of approximately 90 critical Phase-I 

and –II drug-metabolizing enzymes (DMEs) in liver. Starting at 

2-months of age, CV and GF mice (male, n=6-8/group) were 

treated with or without VSL3 (4.5×10 6 CFU/ml) in drinking 

water for 28 days. In a separate study, 1-month old GF mice 

(male, n=4/group) were taken out of the GF isolator and were 

housed in the conventional environment for 2 months. Regarding 

the bacteria profiles, VSL3 increased each VSL3 component 

in the large intestinal content of CV mice, and increased 

these bacteria even more in GF mice, likely due to less competition 

for growth in the GF-environment (16S rRNA qPCR). 

Conventionalization of GF mice increased the total bacteria 

and selected bacteria strains with known important functions 

for host metabolism in the large intestinal content. Regarding 

the effect of VSL3 on the expression of DMEs in livers of CV 

mice, VSL3 increased the mRNAs of the Phase-I enzymes cytochrome 

P450 (Cyp) 4v3, Alcohol dehydrogenase (Adh) 1, 

and Carboxyesterase (Ces) 2a, but decreased the mRNAs 

of Cyp3a44 and 3a11; VSL3 also decreased the mRNAs 

of multiple Phase-II glutathione-S-transferases (Gstm1-3 and 

Gsto1). In livers of GF mice, VSL3 decreased the mRNAs of 

the UDP-glucuronosyl transferases (Ugt) 1a9 and 2a3. Regarding 

the effects of GF and conventionalization on the hepatic 

expression of DMEs, GF mouse livers had increased mRNA of 

Ugt1a9, but decreased mRNAs of Gstpi and sulfotransferase 

5a1. GF conditions down-regulated many genes in the Cyp3a 

cluster but up-regulated many genes in the Cyp4a cluster. Conventionalization 

normalized the expression of these genes back 

to CV levels. The protein expression of Cyp3a and Cyp4a was 

confirmed by Western blot. ChIP-qPCR indicated that changes 

in the hepatic PXR- and PPARα-DNA binding to the Cyp3a 

and Cyp4a gene clusters during GF- and exGF-conditions 

appeared to be responsible for the changes in the Cyp3a and 

4a gene expression. In conclusion, alterations in the intestinal 

bacteria by VSL3, GF condition, and conventionalization 

impact the expression of many DMEs in the host liver, suggesting 

the importance of considering “bacteria-drug” interactions 

in human populations. (Supported by NIH GM111381, 

ES019487, and P30 ES0007033) 

Disclosures: 

The following authors have nothing to disclose: Felcy pavithra Selwyn samraj, 

Sunny Lihua Cheng, Curtis Klaassen, Julia Yue Cui 

656 

HMGB1 interacts with AIM2 to upregulate hepatocyte 

autophagy and prevent cell death after redox stress 

Qian Sun, Timothy R. Billiar, Melanie Scott; Surgery, university of 

pittsburgh, Pittsburgh, PA 

The release of damage-associated molecular patterns such as 

double-stranded DNA (dsDNA) after oxidative stress leads 

to maturation of the inflammasome and caspase-1 in both 

immune and non-immune cells. We have previously shown 

that activation of AIM2 inflammasome and caspase-1 in 

mouse hepatocytes (HCs) upregulates mitochondrial autophagy 

after redox stress rather than leading to the maturation of 

cytokines as in immune cells. Nuclear protein HMGB1 binds 

dsDNA and translocates to the cytosol during redox stress. 

Cytosolic HMGB1 has been shown to upregulate autophagy 

to protect against mitochondrial dysfunction. Therefore, we 

hypothesized that cytosolic HMGB1 interacts with AIM2, and 

its activating ligand dsDNA, to regulate caspase-1 activation 

and caspase-1-mediated mitochondrial autophagy in HCs. 

Methods: HCs isolated from C57BL/6 (WT) and HC-HMGB1 -/- 

(hepatocyte-specific HMGB1 knockout) mice were transfected 

with GFP-LC3 before hypoxia (6h, 1% O 2 

) and reoxygenation 

treatment. Levels of autophagic flux were assessed by increase 

in the number of GFP–LC3 puncta in HCs after treatment of 

bafilomycin (50nM, 1h). Mitochondrial and cytosolic specific 

ROS production was measured by HyPer-Mito and HyPer- 

Cyto respectively. Cell death was determined by Annexin V/ 

PI staining. Interaction between HMGB1 and AIM2 in HCs 

was assessed by immunoprecipitation. Results: Hypoxia/reoxygenation 

triggered enhanced association between AIM2 and 

HMGB1, as well as increased caspase-1 activity in WT HCs, 

but not in HC-HMGB1 -/- cells, suggesting a role for HMGB1 

in regulating caspase-1 activation. Autophagy levels were 

increased in WT HCs after hypoxia/reoxygenation as shown 

by increased beclin1 expression and autophagic flux. However, 

HC-HMGB1 -/- HCs showed decreased beclin1 levels compared 

with WT and impaired autophagic flux after hypoxia/ 

reoxygenation, similar to our previous results in AIM2 -/- HCs. 

Additionally, HC-HMGB1 -/- HCs had increased mitochondrial 

volume and ROS production in the mitochondria in comparison 

with WT after hypoxia/reoxygenation, suggesting a role for 

HMGB1 in up-regulating mitochondrial autophagy. Consistent 

with our previous findings in caspase-1 -/- and AIM2 -/- HCs, 

HC-HMGB1 -/- HCs showed increased apoptosis and necrosis 

in comparison with WT after hypoxia/reoxygenation, confirming 

a protective role for HMGB1 in HCs. Conclusion: Our data 

suggest that HMGB1 is protective in HCs after redox stress by 

upregulating mitochondrial autophagy through its interaction 

with AIM2. Our study links HMGB1 with AIM2 inflammasome 

activation and stress-induced autophagy in non-immune cells, 

which may have implications for future inflammasome-targeting 

therapies. 

Disclosures: 

The following authors have nothing to disclose: Qian Sun, Timothy R. Billiar, 

Melanie Scott 

657 

MYCN is A Novel Biomarker for Chemoprevention of 

Hepatocellular Carcinoma by Acyclic Retinoid 

Soichi Kojima 1 , Xian-Yang Qin 1 , Harukazu Suzuki 2 , Masao 

Honda 3 , Goshi Shiota 4 , Naoto Ishibashi 5 , Hisataka Moriwaki 6 , 

Masahito Shimizu 7 ; 1 Micro-Signaling Regulation Technology 

Unit, RIKEN, Wako, Japan; 2 Division of Genomic Technologies, 

RIKEN CLST, Yokohama, Japan; 3 Department of Gastroenterology, 

Kanazawa University, Kanazawa, Japan; 4 Division of Molecular 

and Genetic Medicine, Tottori University, Yonago, Japan; 5 Pharmaceutical 

Division, KOWA Company, Higashimurayama, Japan; 

6 Gifu University, Gifu, Japan; 7 Department of Gastroenterology, 

Gifu University School of Medicine, Gifu, Japan 

Background & aim: Poor prognosis of hepatocellular carcinoma 

(HCC) is partly due to its high rate of recurrence. Acyclic 

retinoid (ACR) is under phase III clinical trials in Japan to


prevent the recurrence and development of HCC after surgical 

removal of the primary tumors. Previous in vitro experiments 

showed that ACR selectively suppresses the growth of human 

HCC cells (JHH7, HuH7, and HepG2) but not normal human 

hepatic cells (Hc). In the present study, the genome-wide transcriptome 

analysis was performed to identify a biomarker for 

ACR treatment of HCC. Method: Differential gene expression 

profiles of JHH7 and Hc cells treated with ACR were measured 

using the next-generation sequencing-based Cap Analysis 

Gene Expression (CAGE) analysis. Inhibitory effect of ACR on 

MYCN expression was confirmed by real-time PCR in culture 

cells, animal models and liver biopsy of HCC patients administered 

with ACR (n = 6). MYCN-dependent signaling pathways 

underlying the growth suppression by ACR were explored 

using knowledge-based ingenuity pathway analysis (IPA), 

RNA interference, chemical inhibitors and luciferase assays. 

Relationship between MYCN levels and tumor recurrence in 

HCC patients (n = 102) was evaluated using the log-rank test. 

Correlations between MYCN and liver cancer stem cell (CSC) 

markers were determined using Pearson correlation coefficients 

based on microarray database or in patients. Results: MYCN 

was expressed in JHH7 cells but not in Hc cells, and inhibited 

by ACR but not by its ethyl analogues that showed no growth 

suppression. ACR also selectively suppressed the growth of 

MYCN-positive neuroblastoma cells, but not that negative for 

MYCN. Four out of 6 liver biopsies of HCC patients (66.7%) 

who had received 8 weeks of ACR (600 mg/day) after definitive 

treatment showed decreased MYCN expression (< 0.5- 

fold). The result of IPA suggested a Sp1/MYCN/caspase-8 

signaling pathway underlying ACR’s anticancer effect. Upon 

knockdown of MYCN, HCC cells showed suppressed cell 

growth and increased caspase-8 activity. An Sp1 inhibitor 

suppressed MYCN expression, whereas a caspase-8 inhibitor 

restored ACR-induced growth suppression. Finally, clinical 

analysis indicated that MYCN expression in HCC tumors 

was significantly correlated with CSC markers AFP, EpCAM 

and CD133 but not CD90, and negatively associated with 

recurrence of early-stage HCC. Conclusion: MYCN is a new 

biomarker of eligibility for ACR treatment that correlates with 

liver CSC with progenitor features and poor prognosis of HCC. 

ACR suppressed HCC cell growth through a Sp1/MYCN/ 

caspase-8 dependent signaling pathway. 

Disclosures: 

Naoto Ishibashi - Employment: KOWA Company, LTD. 

The following authors have nothing to disclose: Soichi Kojima, Xian-Yang Qin, 

Harukazu Suzuki, Masao Honda, Goshi Shiota, Hisataka Moriwaki, Masahito 

Shimizu 

658 

Farnesoid X Receptor (FXR) Activation by Bile Salts and 

FXR Agonists Increases the Hepatic Expression of the 

Unfolded Protein Response Regulator X-box Binding 

Protein 1 Spliced (XBP1s) 

Xiaoying Liu, David Hilburn, Seong W. Park, Brian E. LeCuyer, 

Richard M. Green; Northwestern University, Chicago, IL 

The Unfolded Protein Response (UPR) is an adaptive response 

to endoplasmic reticulum stress that occurs in cholestatic and 

other forms of liver disease. X-box binding protein 1 spliced 

(XBP1s) is a highly conserved regulator of the UPR, and is 

formed by an unconventional splicing mechanism that removes 

26 nucleotides from the unspliced XBP1 mRNA. The farnesoid 

X receptor (FXR) is a nuclear bile acid receptor which regulates 

bile acid metabolism, and FXR agonists may provide 

novel therapies for cholestatic and fatty liver diseases. In this 

study, we demonstrate a novel role of FXR in regulating liver 

XBP1 splicing and XBP1s expression. Methods: Male FVB mice 

(8-12 weeks) were fed chow supplemented with 0.3% sodium 

deoxycholate (DCA), an endogenous FXR bile salt ligand, for 

up to 7 days. Huh7/NTCP or HepG2 cells were treated with 

either bile salts (GCDCA, TCDCA, TDCA and TUDCA) or the 

synthetic FXR agonists obeticholic acid (OCA) and GW4064 

for 4-8 hours. These cells were also co- treated with the FXR 

antagonist guggulsterone, and either FXR siRNA or control 

siRNA. Gene and protein expression was analyzed by qPCR 

and Western blotting. XBP1 splicing activity was measured 

using an XBP1-luciferase reporter construct that assays the 26-nt 

atypical splicing of the XBP1 mRNA. Results: Hepatic XBP1s 

and its downstream target ERdj4 mRNA expression increased 

6.5-fold (P < 0.05) and 1.7-fold (P < 0.05) in mouse livers after 

DCA feeding for 1 and 3 days, respectively. GCDCA (50 μM), 

TCDCA (10 μM) and TDCA (10 μM) treatment increased XBP1s 

protein expression in Huh7/NTCP cells, while TUDCA (10 

μM) had no effect. The synthetic FXR agonists OCA (2.5 μM) 

and GW4064 (2.5 μM) also induced XBP1s mRNA and protein 

expression. XBP1s protein induction by FXR agonists was 

blocked by the FXR antagonist guggulsterone (50 μM) and by 

FXR siRNA. The FXR agonists TCDCA (100 μM) and GW4064 

(10 μM) also increased XBP1 splicing-luciferase activity 2 and 

4-fold, respectively compared to controls (P < 0.001). Conclusions: 

DCA bile salt feeding to mice increases the expression 

of hepatic XBP1s and its downstream target ERdj4. FXR bile 

salt ligands and synthetic agonists increase XBP1s expression 

in Huh7/NTCP or HepG2 cells, while TUDCA has no effect. 

These effects are due, at least in part, to enhanced XBP1 splicing 

into XBP1s. Increased XBP1s expression is blocked by the 

FXR antagonist guggulsterone and by FXR knockdown. We 

demonstrate the novel finding that a FXR-mediated pathway 

increases the expression of liver XBP1s. We speculate that this 

may be important in the pathogenesis of cholestatic and fatty 

liver diseases. 

Disclosures: 

Richard M. Green - Consulting: McNeil 

The following authors have nothing to disclose: Xiaoying Liu, David Hilburn, 

Seong W. Park, Brian E. LeCuyer 

659 

Cerium oxide nanoparticle treatment enhances liver 

regeneration after two-thirds partial hepatectomy in 

rats 

Manuel Morales-Ruiz 1,3 , Altamira Arce-Cerezo 1,5 , Montserrat 

Pauta 1,5 , Jordi Ribera 1,3 , Denise Oró 1,3 , Gregori Casals 1,3 , Guillermo 

Fernández-Varo 1,3 , Tetyana Yudina 4 , Víctor Puntes 4 , Wladimiro 

Jiménez 1,2 ; 1 Biochemistry and Molecular Genetics, Hospital 

Clinic, Barcelona, Spain; 2 Department of Physiological Sciences I, 

IDIBAPS, CIBERehd, University of Barcelona School of Medicine, 

Barcelona, Spain; 3 IDIBAPS, CIBERehd, Barcelona, Spain; 4 Catalan 

Institute of Nanoscience and Nanotechnology, UAB, Bellaterra, 

Spain; 5 IDIBAPS, Barcelona, Spain 

Introduction and aim: Several studies have shown that oxidative 

stress impairs hepatic regeneration in mice. Therefore, 

testing new anti-oxidant drugs to improve liver regeneration 

has clinical interest. Recently, considerable attention has been 

paid to cerium oxide nanoparticles (CeO 2 

NPs) as a treatment 

for oxidative stress-related diseases. This interest relies on the 

multi-enzyme mimetic properties of CeO 2 

NPs that turn these 

nanoparticles into an effective scavenger of reactive oxygen 

species. Consequently, we aimed to investigate the effect of 

CeO 2 

NPs treatment on hepatic regeneration after partial hepatectomy. 

Methods: CeO 2 

NPs (4 nm in size), were synthesized 

by precipitation of a cerium (III) salt by the addition of ammo-


nium salt tetramethylammonium-hydroxide (TmaOH). Forty-four 

male Wistar rats were divided equally into two groups. In the 

first group, rats were treated with 0.1 mg/kg CeO 2 

NPs i.v. 

twice a week for two weeks before 2/3 partial hepatectomy 

(PHx). The second group received vehicle as a control treatment. 

PHx was performed in both experimental groups and 

rats were sacrificed at different time points for further analysis. 

Results: Rats were sacrificed at day 6 after PHx and the ratio 

“liver remnant weight/total body weight” was calculated to 

obtain the liver regenerative index. Rats treated with CeO 2 

NPs 

showed a significant 11% increase in liver regeneration, compared 

with vehicle treated rats (p


Disclosures: 

The following authors have nothing to disclose: Emma A. Kruglov, Meena Ananthanarayanan, 

Jittima Weerachayaphorn, Pedro Sousa, Michael H. Nathanson 

662 

Effects of Ursodeoxycholic-acid and Rifampicin on autophagy 

in the liver 

Katrin Panzitt 1 , Hanns-Ulrich Marschall 2 , Michael Trauner 3 , Peter 

Fickert 1 , Martin Wagner 1 ; 1 Division of Gastroenterology and 

Hepatology, Department of Internal Medicine, Medical University 

Graz, Graz, Austria; 2 Wallenberg Laboratory, Department 

of Molecular and Clinical Medicine, Sahlgrenska Academy, University 

of Gothenburg, Gothenburg, Sweden; 3 Division of Gastroenterology 

and Hepatology, Department of Internal Medicine III, 


Background: Bile acids and activation of the bile acid receptor 

FXR inhibit autophagy, a cellular self-digestion process necessary 

for cell homeostasis and regeneration. The effects of 

chronic bile acid accumulation in chronic cholestatic liver disease 

on autophagy have not been studied in detail. However, 

indirect evidence (e.g. accumulation of Mallory-Denk bodies 

in primary biliary cirrhosis) indicates that autophagy may be 

impaired in human cholestasis. Hypothesis: We aim to determine 

whether ursodeoxycholic-acid (UDCA) and rifampicin 

(Rifa), two drugs for the treatment of human cholestatic liver 

disease may activate autophagy as a potential mode of drug 

action. Methods: Markers of autophagy (LC3, p62, ATG5, 

ATG7, ATG12) and the upstream mTOR signaling pathway 

(Raptor, ULK1, pS6K) have been studied by Western blot, 

immunofluorescence and qPCR in liver biopsies from patients 

treated with UDCA and Rifa. Mechanistic details of UDCA 

and Rifa action have further been studied in detail in human 

HepG2 cells and human primary hepatocytes using shPXR 

and shFXR knockdown strategies as well as conventional techniques 

to study autophagy (e.g. chloroquine treatment, LC3- 

RFP-GFP transfections). Results: Both, UDCA and Rifa induce 

LC3 II protein levels as the main autophagy readout in human 

liver biopsies. More detailed studies in hepatocyte cell lines 

using chloroquine treatment and LC3-RFP-GFP transfections 

reveal that true autophagic flux is induced. UDCA appears 

to activate autophagy via mTOR-ULK1 signaling whereas Rifa 

induces autophagy on transcriptional levels (LC3C, LAMP1, 

ATG10) without impacting on mTOR signaling. Furthermore, 

knockdown of the Rifa activated transcription factor PXR significantly 

represses autophagy already under basal conditions on 

mRNA and protein levels. In addition, PXR knockdown prevents 

Rifa induced autophagy induction. Summary and conclusions: 

UDCA and Rifa induce autophagy in the liver via different 

mechanisms. UDCA induces autophagy fux via mTOR signaling 

pathways whereas Rifa induces autophagy flux mTOR independently 

via the transcription factor PXR. Part of the beneficial 

effects of UDCA and Rifa in the treatment of cholestatic liver 

disease may be attributed to an induction of autophagy. Both 

compounds, UDCA and Rifa may have additional beneficial 

effect by inducing autophagy on other hepatological as well as 

non-hepatological diseases. 

Disclosures: 






Peter Fickert - Consulting: Falk Foundation, Falk Foundation, Falk Foundation, 

Falk Foundation; Speaking and Teaching: Roche Austria, Gilead Austria, MSD 

Austria, MERCK Austria, Roche Austria, Gilead Austria, MSD Austria, MERCK 

Austria, Roche Austria, Gilead Austria, MSD Austria, MERCK Austria, Roche 

Austria, Gilead Austria, MSD Austria, MERCK Austria 

The following authors have nothing to disclose: Katrin Panzitt, Martin Wagner 

663 

Involvement of Endoplasmic Reticulum Stress Transducer 

BBF2H7 in Hepatocellular Carcinoma Promotion 

Yizhou Zhang 1,2 , Nelson Hayes 1,2 , Masataka Tsuge 1,2 , Nobuhiko 

Hiraga 1,2 , Takuro Uchida 1,2 , Daiki Miki 3,2 , Hiromi Abe 1,2 , Kazuaki 

Chayama 1,2 ; 1 Department of Gastroenterology and Metabolism, 

Hiroshima University, Hiroshima City, Japan; 2 Liver Research Project 

Center, Hiroshima University, Hiroshima City, Japan; 3 RIKEN 

Center for Integrative Medical Sciences, Yokohama, Japan 

Objectives: Internal and external environmental stress brings 

about dysregulation of gene expression and genomic instabilities. 

The fundamental role of the tumor suppressor p53 in 

the response to various forms of stress ensures our genomic 

integrity. However, p53 is reported to be functional inactivated 

in almost all cancers. Endoplasmic reticulum (ER) stress 

is indicated as a factor contributing to cancer. So far, how 

effectors of ER stress cause Hepatocellular Carcinoma (HCC) 

is not well studied. Here we show that ER stress transducer 

BBF2H7 promotes ubiquitin-dependent p53 degradation and 

diminishes p53 phosphorylation, which in turn contributes to 

cell proliferation and survival of HCC. Methods: Expression of 

ER stress transducers in paired non-tumor and HCC tissues was 

measured by Real-Time PCR. Immunohistochemistry of HCC 

tissues was performed with BBF2H7 antibody. Cell growth of 

HepG2 cells with BBF2H7 overexpressed or knocked down 

was monitored by live cell imaging. Expression of p53 and 

its downstream targets were checked by Real-Time PCR and 

western blot in BBF2H7 overexpressed HepG2 cells, with or 

without the addition of proteasome inhibitor MG132. Dual 

luciferase reporter assay was performed to test how BBF2H7 

modulates p53 transcriptional activities. Nuclear abundance of 

p53 affected by BBF2H7 was also identified by immunostaining. 

Results: Among all the ER stress transducers, BBF2H7, an 

ERresident basic leucine zipper transcription factor, was found 

to be upregulated in HCC. IHC showed BBF2H7 accumulated 

in the nucleus of tumor cells, which is different from that in 

adjacent non-tumor tissues. Cell growth of HepG2 cells was 

promoted by BBF2H7 overexpression and was suppressed by 

BBF2H7 siRNA. Real-Time PCR results showed that BBF2H7 

overexpression increased MDM2 and BCL-2 expression, but 

p53 mRNA level was not changed. Protein levels of both p53 

and phos-p53 were decreased by BBF2H7 overexpression, 

but only p53 protein could be restored by blocking proteasome 

activity. The anti-apoptosis proteins BCL-2 and BCL-xL 

were up-regulated by BBF2H7, while Pol.δ cofactor PCNA was 

not affected. Strikingly, p53 transcriptional activity was drastically 

reduced by BBF2H7 overexpression. Contrary to the 

case of overall p53 protein, p53 transcriptional activity could 

barely be recovered by proteasome inhibitor. The impairment 

of p53 nuclear abundance by BBF2H7 might be the reason 

behind this. Conclusions: These findings reveal a new mechanism 

in which p53 can be destabilized by ER stress transducer 

BBF2H7, and also imply that BBF2H7 has full potential to trigger 

HCC carcinogenesis through impairing p53 transcriptional 

activity. 

Disclosures: 








The following authors have nothing to disclose: Yizhou Zhang, Nelson Hayes, 

Masataka Tsuge, Nobuhiko Hiraga, Takuro Uchida, Daiki Miki, Hiromi Abe


664 

Restoration of Hepatic Physiological Parameters in vitro 

Influences Proprotein Convertase Subtilisin Kexin type 9 

(PCSK9) and LDL Regulation by Atorvastatin. 

Banumathi K. Cole, David Manka, Brian R. Wamhoff, Brett R. 

Blackman, Ajit Dash; HemoShear, Charlottesville, VA 

Background: Therapeutic targeting of Proprotein Convertase 

Subtilisin Kexin type 9 (PCSK9) in hyperlipidemia is attractive 

due to its ability to regulate hepatocyte LDL receptor levels and 

thereby clearance of circulating LDL cholesterol. Conventional 

hepatocyte models, however, are troubled by rapid dedifferentiation, 

loss of liver phenotype and function in culture, and 

non-translational drug dosing and exposure responses, challenging 

the relevance of these models and the accuracy of 

their biological response. We previously described a primary 

human hepatocyte system that uses liver-derived hemodynamics 

and physiological transport parameters to restore in vivo-like 

phenotype and hormone/drug responses at exposures matching 

physiological/clinical levels. Aims: The goal of this study 

was to validate the differences in regulation of PCSK9 biology 

and LDL uptake in response to atorvastatin, as they relate to differences 

in LDL exposure and physiological and conventional 

model conditions. Methods: Primary human hepatocyte tissues 

were cultured in collagen gel configurations and applied in 3 

conditions; ‘static’ non-perfused state, ‘hemodynamics’ with 

perfusion, or ‘perfusion’ only controls to account for LDL exposure 

effects. Cultures were exposed to 50μg/ml nLDL and atorvastatin 

for up to 48 hours. Endpoints included immunostaining 

for E-cadherin, measurement of intracellular protein levels of 

PCSK9, furin and HNF1α, secreted levels of PCSK9, and LDL 

uptake kinetics. Results: E-cadherin staining patterns revealed 

peripheral localization indicative of differentiated hepatocytes 

under hemodynamics but not in perfused only cultures. We 

found that continuous loading of nLDL, reflective of physiological 

exposure patterns in both the perfused and hemodynamic 

conditions, decreased baseline LDL uptake rates and PCSK9 

secretion relative to static cultures. The expected effect of atorvastatin 

on increasing PCSK9 secretion was noted only in the 

perfused and hemodynamic conditions but lost in static cultures. 

However, atorvastatin increased intracellular PCSK9 and 

LDL uptake in both static and hemodynamic conditions. Hemodynamics, 

but not perfusion alone, was also seen to upregulate 

key proteins, such as furin and HNF1α, that modulate 

PCSK9 expression and activity, which were not impacted by 

atorvastatin treatment. Conclusion: The results highlight the role 

of a combination of physiological milieu and hemodynamic 

parameters in resetting baseline biology and restoring PCSK9 

processing and secretion, which is important in elucidating the 

mechanisms surrounding PCSK9 regulation and the consequent 

development of therapeutic strategies for lowering circulating 

LDL levels. 

Disclosures: 

Banumathi K. Cole - Employment: HemoShear, LLC 






HemoShear LLC 


665 

The regulation of liver volume gain and regeneration-associated 

steatosis through Pten 

Ekaterina Kachaylo 1 , Christoph Tschuor 1 , Nathalie Borgeaud 1 , 

Perparim Limani 1 , Michelangelo Foti 2 , Rolf Graf 1 , Bostjan Humar 1 , 

Pierre A. Clavien 1 ; 1 Visceral and Transplant Surgery, University 

Hospital of Zurich, Zurich, Switzerland; 2 Département de Physiologie 

Cellulaire & Métabolisme, Centre Médical Universitaire, 

Geneva, Switzerland 

Successful liver regeneration is associated with the transient 

accumulation of lipids in hepatocytes, likely for the provision 

of energy to fuel the regenerative process. How this transient 

steatosis is regulated is ill-understood. The loss of Pten from 

liver induces rapid redistribution of peripheral fats into liver. 

Given that Pten also controls the growth-promoting Akt-mTORC 

pathway, Pten may act after resection to facilitate lipid metabolism 

and to drive liver volume gain. To explore this possibility, 

we are analysing inducible, hepatocyte-specific Pten knockout 

mice following hepatectomy. Pten levels were assessed in 

hepatectomized wild-type mice. AlbCre tg/+ PTEN fl/fl and Alb- 

Cre +/+ PTEN fl/fl (control) mice were studied shortly after tamoxifen 

induction. Regeneration was monitored via hepatic weight 

gain, proliferative activity and cyclin levels. Lipid content was 

quantified through a sulpho-phospho-vanillin protocol. In wild 

type mice, Pten levels dropped along with pronounced T389- 

Akt phosphorylation at 16h post resection, coinciding with the 

peak of transient steatosis. In parallel, fatty acid import (Cd36) 

and β-oxidation (Cpt1a, Hadha/b) increased, whilst lipogenesis 

(Acc, Scd, Fasn) decreased. Inducing Pten loss In AlbCre t- 

g/+ PTEN fl/fl mice resulted in spontaneous lipid accumulation 

associated with increased expression of lipogenic enzymes 

and elevation of serum triglycerides. However already at 16h 

after hepatectomy expression levels of Acc, Scd and Fasn and 

serum triglycerides were similar to controls and 3 days after 

resection, lipid content was comparable to controls, yet liver 

weight gain was accelerated. Notably, the increased liver 

weight gain was not accompanied by elevations in proliferative 

markers, suggesting an involvement of hypertrophy. Western 

blot analysis revealed increased S474-Akt2 phosphorylation at 

32 hours after hepatectomy in Pten ko livers vs controls, which 

indicates changes in glucose and lipid metabolism upon loss of 

Pten. Our results demonstrate a growth-promoting function for 

Pten downregulation during liver regeneration. The reduction 

in Pten may aid liver growth through (i) the release of inhibition 

of the hypertrophic Akt-mTORC pathway, and through (ii) the 

provision of energy via modulation of metabolism. 

Disclosures: 

The following authors have nothing to disclose: Ekaterina Kachaylo, Christoph 

Tschuor, Nathalie Borgeaud, Perparim Limani, Michelangelo Foti, Rolf Graf, 

Bostjan Humar, Pierre A. Clavien 

666 

Organopollutant Exposures Further Decrease Epidermal 

Growth Factor Signaling in Non-Alcoholic Steatohepatitis 

and Alter Hepatic Energy Metabolism 

Josiah Hardesty 2 , Keith C. Falkner 1 , Heather B. Clair 2 , Banrida 

Wahlang 1 , Russell A. Prough 2 , Matthew C. Cave 1,3 ; 1 Department 

of Medicine/GI, University of Louisville, Louisville, KY; 2 Biochemistry, 

University of Louisville, Louisville, KY; 3 Robley Rex VAMC, 

Louisville, KY 

Background: Decreased Epidermal Growth Factor Receptor 

(EGFR) signaling has been associated with non-alcoholic steatohepatitis 

(NASH). We have previously demonstrated that 

the ubiquitous environmental pollutants, polychlorinated biphenyls 

(PCBs), are a ‘second hit’ in the transition of steatosis to


steatohepatitis. PCB-induced nuclear receptor activation only 

partially explained these findings, although the other ‘off target’ 

mechanisms involved are unknown. These observations 

taken together led us to investigate the hypothesis that PCB-mediated 

EGFR inhibition alters downstream targets including the 

mechanistic target of rapamycin (mTOR) – a master regulator 

of hepatic energy metabolism in NASH. Methods: Mice 

were fed either a control diet (CD) or 42% milk fat high fat 

diet (HFD) and treated with either corn oil or Aroclor1260 

(PCB mixture, 20 mg/kg by one time gavage) for 12 weeks. 

Livers were removed and lysed for western blot analysis of 

pEGFR Y1173, EGFR, pmTOR, mTOR, and β-actin. In vitro 

cell culture assays were conducted with HepG2 and AML-12 

cells. Cells were treated with either EGF and EGFR inhibitor, 

or EGF alone, or in combination with Aroclor 1260; proteins 

were extracted for western blot analysis. Results: CD fed mice 

did not develop either steatosis or hepatic necro-inflammation. 

While HFD treated mice developed steatosis, only those co-exposed 

to HFD and Aroclor 1260 developed hepatic inflammation 

and steatohepatitis consistent with NASH. Co-exposed 

mice also had significant alterations in hepatic glucose and 

lipid metabolism. Analysis of liver samples showed an impact 

of diet on EGFR expression in which HFD decreased EGFR 

expression relative to CD. Aroclor-exposed mice demonstrated 

decreased phosphorylated EGFR at Y1173 independent of 

diet. Mice exposed to PCBs also had decreased phosphorylated 

mTOR and decreased total mTOR expression. In HepG2 

or AML-12 cells, Aroclor 1260 exposure was found to diminish 

EGFR phosphorylation at Y1173 relative to the positive control. 

Conclusion: These data demonstrate that HFD can lead 

to decreased hepatic EGFR expression and organopollutant 

exposure can act as a ‘second hit’ further decreasing EGFR 

signaling and altering mTOR activity. These may be the first 

data on environmental chemicals altering tyrosine kinase signaling 

in NASH. These findings require further study but may 

represent a new mechanism by which environmental pollution 

may contribute to NASH and the impaired liver regeneration 

associated with steatosis. Because mTOR inhibition is generally 

anti-inflammatory, additional mechanisms of PCB-induced 

hepatic inflammation should be investigated. 

Disclosures: 

Matthew C. Cave - Advisory Committees or Review Panels: Intercept, Abbvie; 

Consulting: Abbvie, Diapharma; Grant/Research Support: Merck, Gilead, Intercept, 

Conatus, Lumena, Cepheid, Tobira, Galectin, Bayer; Speaking and Teaching: 

BMS, Abbvie, Gilead, Janssen, Genentech 

The following authors have nothing to disclose: Josiah Hardesty, Keith C. Falkner, 

Heather B. Clair, Banrida Wahlang, Russell A. Prough 

667 

Skeletal muscle hyperammonemia causes mitochondrial 

functional abnormalities in cirrhosis 

Gangarao Davuluri 2 , Allawy Allawy 2,6 , Samjhana Thapaliya 2 , 

Dharmvir Singh 2 , Avinash Kumar 2 , Julie H. Rennison 2 , Rafaella 

Nascimento e Silva 2 , Cynthia Tsien 3 , David R. Van Wagoner 4 , 

Sathyamangla V. Naga Prasad 4 , Hoppel Charles 5 , Takhar Kasumov 

2 , Srinivasan Dasarathy 1 ; 1 Department Of Gastroenterology 

and Hepatology, Cleveland Clinic, Cleveland, OH; 2 Pathobiology, 

Cleveland Clinic, Cleveland, OH; 3 Gastroenterology, Toronto 

General Hospital, Toronto, ON, Canada; 4 Molecular Cardiology, 

Cleveland Clinic, Cleveland, OH; 5 Pharmacology and Medicine, 

Case Western Reserve University, Cleveland, OH; 6 Department Of 

Internal Medicine, Cleveland Clinic, Cleveland, OH 

Background. Skeletal muscle is a recognized metabolic partner 

to the liver for ammonia disposal and tissue specific adaptive 

responses occur during hyperammonemia. However, it is 

not known if increased metabolic and cellular stress induced 

by elevated ammonia uptake by the skeletal muscle results in 

defects in mitochondrial bioenergetic function. Methods. Skeletal 

muscle from human cirrhosis and controls, hyperammonemic 

portacaval anastomosis (PCA) rat and sham operated controls 

and myotubes exposed to hyperammonemia were used. Flow 

cytometry, using the fluorophores DCFDA and MitoSox, was 

used to measure reactive oxygen species (ROS) in C2C12 

myotubes during hyperammonemia. Oxidative modification of 

proteins by immublots for carbonylated proteins, lipid peroxidation 

by measuring thiobarbituric acid reactive substances 

(TBARS) and ATP content by fluorometric assays. The site of 

electron leak that generated the ROS was determined my quantifying 

H 2 

O 2 

production by oxidation of fluorogenic indicator 

amplexred in the presence of specific inhibitors (rotenone, 

malonate, antimycin A,oligomycin) of the electron transport 

chain. Cellular NAD+ and NADH were quantified by a fluorometric 

assay with and without inhibitors of the ETC to determine 

the mechanism of impaired electron flow. Results. Hyperammonemia 

increased mitochondrial ROS in C2C12 myotubes 

that was reversed by MitoTEMPO, a specific mitochondrial 

ROS scavenger. ATP content was lower, and carbonylated 

proteins and TBARS were significantly higher in the skeletal 

muscle from patients with cirrhosis and the PCA rat and in 

myotubes during hyperammonemia compared to the respective 

controls. Lowering cellular ammonia concentration reversed 

the low ATP content observed during hyperammonemia. Using 

sequential blockers of the various electron transport chain complexes, 

we demonstrate that Complex III was the major source 

of electron leak and ROS production production in the skeletal 

muscle during hyperammonemia. Finally, NAD+/NADH ratio 

was decreased with lower NAD+ and increased NADH due to 

impaired Complex I of the ETC NADH oxidase during hyperammonemia. 

Conclusions. Our studies in a comprehensive 

array of models show that skeletal muscle ammonia disposal 

activates a sequence of metabolic responses that culminate in 

disordered mitochondrial function. Notably, the reduction in 

ATP content is accompanied by electron leak at complex III 

and consequent generation of ROS during hyperammonemia. 

Reduction in ammonia can potentially reverse the mitochondrial 

bioenergetics dysfunction. These data provide evidence of 

a novel mechanism of hyperammonemia-induced perturbations 

in skeletal muscle cellular bioenergetics. 

Disclosures: 

The following authors have nothing to disclose: Gangarao Davuluri, Allawy 

Allawy, Samjhana Thapaliya, Dharmvir Singh, Avinash Kumar, Julie H. Rennison, 

Rafaella Nascimento e Silva, Cynthia Tsien, David R. Van Wagoner, 

Sathyamangla V. Naga Prasad, Hoppel Charles, Takhar Kasumov, Srinivasan 

Dasarathy 

668 

Hyperammonemia activates a leucine responsive amino 

acid starvation stress response in the skeletal muscle 

Gangarao Davuluri 2 , Dawid Krokowski 3 , Bo-Jhih Guan 3 , Dharmvir 

Singh 2 , Allawy Allawy 2,6 , Avinash Kumar 2 , Rafaella Nascimento e 

Silva 2 , Ashok Runkana 2 , Samjhana Thapaliya 2 , Chenyang Zhao 4 , 

Thomas Hamilton 4 , Sathyamangla V. Naga Prasad 5 , Maria Hatzoglou 

3 , Srinivasan Dasarathy 1 ; 1 Department Of Gastroenterology 

and Hepatology, Cleveland Clinic, Cleveland, OH; 2 Pathobiology, 

Cleveland Clinic, Cleveland, OH; 3 Department of Pharmacology, 

Case Western Reserve University, Cleveland, OH; 4 Immunology, 

Cleveland Clinic, Cleveland, OH; 5 Molecular Cardiology, Cleveland 

Clinic, Cleveland, OH; 6 Department Of Internal Medicine, 

Cleveland Clinic, Cleveland, OH 

Backgound. Skeletal muscle ammonia concentrations are 

increased in cirrhosis and results in sarcopenia. Decreased 

plasma and muscle concentrations of leucine and increased


plasma concentration of glutamine accompany cirrhosis. Here 

we show that hyperammonemia activates a specific molecular 

program in the skeletal muscle increasing the cellular demand 

for leucine that is transported into the mitochondria to potentially 

serve as a metabolic substrate for glutamine synthesis. 

Methods. Studies were performed in the skeletal muscle from 

patients with cirrhosis, hyperammonemic portacaval anastomosis 

rat, differentiated murine C2C12 myotubes during hyperammonemia 

appropriate controls. Myotubes were exposed 

to either ammonium acetate (10mM) or leucine (5mM) or a 

combination of leucine and ammonium acetate. Expression of 

mTOR, its target signaling proteins, global repressor of protein 

synthesis eIF2α and intracellular amino acid deficiency sensor, 

GCN2 expression were quantified by immunoblots. System L 

leucine transporter (LAT1) and glultamine transporter SLC38A2 

were quantified by real time PCR. Rate of protein synthesis was 

quantified using 3 H phenylalanine incorporation. Cellular and 

mitochondrial leucine uptake in myotubes was quantified using 

3 H leucine uptake and amino acids quantified using HPLC. 

Results. Skeletal muscle from cirrhotic patients and the PCA 

rat showed increased phosphorylation of eIF2α and reduced 

mTOR activation compared to that from respective controls. 

Murine myotubes showed activation of general control derepressed-2 

(GCN2) and its target, eIF2α, a global repressor of 

protein synthesis during hyperammonemia. This was accompanied 

by impaired mTOR signaling and reduced protein synthesis. 

Both the molecular perturbations and impaired protein 

synthesis were reversed by L-leucine supplementation in the 

medium. Lack of induction of GADD34 expression, an eIF2α 

phosphatase during hyperammonemia resulted in prolonged 

phosphorylation and activation of eIF2α. Increased expression 

of leucine exchanger, LAT1 but not glutamine transporter, 

SLC38A2 in both human muscle and murine myotubes were 

observed during hyperammonemia. Conclusions. We demonstrate 

a novel amino acid starvation response in the skeletal 

muscle during hyperammonemia that responds specifically to 

leucine supplementation despite increased leucine transport 

and intracellular concentrations. These observations combined 

with mitochondrial transport of leucine suggest that increased 

demand due to cataplerosis results in increased mitochondrial 

transport and activation of a specific program of translational 

repression mediated by persistent activation of eIF2α and 

impaired mTOR1 signaling. 

Disclosures: 

The following authors have nothing to disclose: Gangarao Davuluri, Dawid Krokowski, 

Bo-Jhih Guan, Dharmvir Singh, Allawy Allawy, Avinash Kumar, Rafaella 

Nascimento e Silva, Ashok Runkana, Samjhana Thapaliya, Chenyang Zhao, 

Thomas Hamilton, Sathyamangla V. Naga Prasad, Maria Hatzoglou, Srinivasan 

Dasarathy 

669 

Orchestration of BH3-only proteins Bim, Bid and Puma 

controls hepatocyte apoptosis in Bcl-xL/Mcl-1 knockout 

mice 

Yoshinobu Saito, Hayato Hikita, Takahiro Kodama, Yuto Shiode, 

Yasutoshi Nozaki, Yugo Kai, Yuki Makino, Tasuku Nakabori, 

Satoshi Tanaka, Satoshi Aono, Ryotaro Sakamori, Naoki Hiramatsu, 

Tomohide Tatsumi, Tetsuo Takehara; Gastroenterology 

and Hepatology, Osaka University Graduate School of Medicine, 

Suita, Japan 

Background and Aim: Liver humanized mouse models using 

Alb-uPA SCID mice and Alb-HSVtk NOG mice have been 

reported to provide a lot of information on human liver physiology 

and pathology. However, these mouse models are not 

appropriate for investigating chronic inflammation and liver 

cancer because these mice are immune-deficient and could not 

survive more than one year. Immune tolerance is considered 

to be established during embryonic or neonatal stage. In this 

study, we investigated the engraftment efficacy of allogeneic or 

xenogeneic hepatocytes into the fetal liver by in utero transplantation 

in immune-competent mice. Method: As immune-competent 

recipient mice, we used hepatocyte-specific Mcl-1 knockout 

(KO) mice, which cause spontaneous hepatocyte apoptosis 

in life (Liver Injury mice), and hepatocyte-specific Mcl-1 and 

Bcl-xL double KO (DKO) mice, which show a decreased number 

of hepatocytes on embryonic (E) 18.5 day and die within 

one day after birth due to hepatic failure (Liver Impairment 

mice). Primary hepatocytes isolated from ubiquitously green fluorescent 

protein (GFP) transgenic (Tg) mice or primary human 

hepatocytes were administered into these recipient mice on E 

16.5 day via vitelline vein. These embryos were given birth by 

cesarean section and these livers were investigated at birth or 

at the age of 6 weeks. Results: Hepatocyte-transplanted wildtype 

mice and Liver Injury mice showed scattered embolization 

areas at birth by HE staining of the liver sections. The embolization 

was considered to be caused by transplanted hepatocytes. 

In contrast, Liver Impairment mouse livers escaped from 

embolization. In livers of wild-type mice and Liver Injury mice 

transplanted with GFP Tg hepatocytes, GFP-positive cells were 

observed forming a lot of clusters. While repopulation rate was 

not different between livers of wild-type mice and those of Liver 

Injury mice at birth (35.8% (3.6-48.3) and 29.8% (20.4-39.1), 

respectively), that of Liver Impairment was 7.8% (3.3-16.7). 

Meanwhile, repopulation rate at the age of 6 weeks was 0% in 

wild-type mice and 4.2% (2.1-8.0) in Liver Injury mice. In livers 

of wild-type and Liver Injury mice transplanted with primary 

human hepatocytes, embolization was similarly observed at 

birth by HE staining, suggesting that transplanted hepatocytes 

might be engrafted. Conclusion: Our results suggests that transplantation 

of primary hepatocytes into the fetal liver of Liver 

Injury mice, but not Liver Impairment mice, have the possibility 

of generating humanized liver mice without immunodeficiency. 

Disclosures: 






The following authors have nothing to disclose: Yoshinobu Saito, Takahiro 

Kodama, Yuto Shiode, Yasutoshi Nozaki, Yugo Kai, Yuki Makino, Tasuku Nakabori, 

Satoshi Tanaka, Satoshi Aono, Ryotaro Sakamori, Naoki Hiramatsu, 

Tomohide Tatsumi 

670 

Sustained Inhibition of Liver Regeneration in Mice following 

Combined Elimination of MET & EGFR Signaling 

Shirish Paranjpe, William C. Bowen, Meagan Haynes, Anne Orr, 

Wendy M. Mars, Jianhua Luo, George K. Michalopoulos; Pathology, 

University of Pittsburgh, Pittsburgh, PA 

Acute loss of liver mass following toxic or surgical means elicits 

hepatic regeneration and restoration of liver tissue to its original 

mass, indicating a tightly regulated growth process. PHx 

in mice and rats results in rapid induction of more than 100 

genes, (that are not expressed in resting liver) & activation 

of multiple signaling pathways. EGFR & MET, the two major 

mitogenic receptor tyrosine kinases, are activated within 15 - 

30 minutes following a PHx. The MET-EGFR signaling pathway 

plays a significant role in is activated within 15-30 minute 

following a partial hepatectomy (PHx) in mice and rats. The 

role played by these two pathways during liver regeneration 

was investigated by utilizing mice that had c-met deleted in the 

liver using a tamoxiphen inducible Cre-loxP system. Mice were


injected with Tamoxiphen (1mg/ml) and injected i.p everyday 

for 5 days. Controls were injected with corn oil alone.This 

treatment deleted exon 16 that contains a critical ATP-binding 

site in the intracellular tyrosine kinase (TK) domain, essential for 

the activation of c-met. PhosphoEGFR was inhibited by using 

Canertinib, an irreversible pan-pEGFR inhibitor that was administered 

i.p at 80 mg/kg. Mice were injected a day before PHx 

and then everyday for 14 days. Appropriate vehicle controls 

were also used. In mice treated with Tyrosine kinase inhibitors, 

pEGFR levels were significantly reduced compared to vehicle 

treated controls. Analysis of proliferation indices indicated significant 

drop in KI67 positive staining in treated mice compared 

to controls Conclusion: Combined elimination of both MET and 

EGFR signaling resulted in sustained inhibition of liver regeneration 

up to 14 days.The PI3K-AKT-mTOR and the RAS-MEK- 

ERK pathways associated with cellular proliferation were down 

regulated in MET-EGFR inhibited mice.A significant reduction 

in Ki67 labeling was also evident, as was a reduction in liver to 

body weight ratio, suggesting a liver regeneration deficit.The 

increase in hepatocyte size in Met and EGFR suppressed livers 

may represent a compensatory effect to increase liver weight 

in the absence of hepatocyte proliferation.Suppression of MET- 

EGFR caused profound alterations in expression of many cell 

cycle associated genes. microRNA`s and LncRNA`s. 

Disclosures: 

George K. Michalopoulos - Consulting: Vital Therapies 

The following authors have nothing to disclose: Shirish Paranjpe, William C. 

Bowen, Meagan Haynes, Anne Orr, Wendy M. Mars, Jianhua Luo 

671 

Hyperammonemia impairs skeletal muscle protein synthesis 

via a myostatin dependent mechanism 

Srinivasan Dasarathy 1 , Gangarao Davuluri 2 , Samjhana Thapaliya 

2 , Avinash Kumar 2 , Gabriella A. Ten Have 3 , Dharmvir 

Singh 2 , Stephen L. Welle 4 , Marielle Engelen 3 , Sathyamangla 

V. Naga Prasad 5 , Nicolaas E. Deutz 3 ; 1 Department Of Gastroenterology 

and Hepatology, Cleveland Clinic, Cleveland, OH; 

2 Pathobiology, Cleveland Clinic, Cleveland, OH; 3 Department of 

Health and Kinesiology, Texas A&M University, College Station, 

TX; 4 Medicine, University of Rochester Medical Center, Rochester, 

NY; 5 Molecular Cardiology, Cleveland Clinic, Cleveland, OH 

Hyperammonemia is a consistent abnormality in cirrhosis and 

contributes to sarcopenia or skeletal muscle loss. We have 

previously shown that myostatin expression is increased in 

the hyperammonemic portacaval anastomosis (PCA) rat and 

in C2C12 myotubes in vitro. To demonstrate that hyperammonemia 

induced skeletal muscle loss is mediated via an 

increased expression of myostatin in vivo, we induced hyperammonemia 

in post developmental myostatin-/- and myostatin+/+ 

mice and the metabolic and molecular signaling 

responses regulating protein synthesis (mTOR1 targets p70s6k, 

ribosomal s6 protein, 4EBP1) in the gastrocnemius muscle were 

studied. The post developmental myostatin knockout mice were 

generated by floxing the 3rd exon of myostatin gene (codes 

for entire active protein), a transgene encoding tamoxifen-activated 

Cre recombinase. At 4 months of age, mice were fed 

tamoxifen for 2 weeks to deplete myostatin. Hyperammonemia 

was induced by daily intraperitoneal injection of ammonium 

acetate for 4 weeks. Muscle protein synthesis was quantified 

by administration of a pulse of L-ring-D5 phenylalanine 

(D5Phe) and protein incorporation of labeled D5-Phe quantified 

using LC-MS/MS. Plasma (60.1±22.2 vs 378.0±71.2 

μmol/l; p


673 

Effects of Targeting Wnt/β-Catenin and RAS/RAF/ 

MAPK Pathway on Hepatocellular Carcinoma Cell 

Growth and Metabolism: Potential for a New Combination 

Therapy 

Lilia Turcios 1 , Valery Vilchez 1 , David Butterfield 2 , Mihail I. Mitov 3 , 

Francesc Marti 1 , Roberto Gedaly 1 ; 1 Surgery, Univ of Kentucky, 

Lexington, KY; 2 Chemistry, University of Kentucky, Lexington, KY; 

3 Markey Cancer Center - Core Support, University of Kentucky, 

Lexington, KY 

Background: Treatment of advanced hepatocellular carcinoma 

(HCC) remains a challenge. We aim to evaluate the impact 

of a β-catenin inhibitor, FH535, alone or in combination with 

a RAS/RAF/MAPK inhibitor, sorafenib, in liver cancer cells. 

Methods: Proliferation of liver cancer cell lines (Huh7, PLC, 

Hep3B) with or without addition of FH535 (10mM, 5mM, 

2.5mM) and sorafenib (2mM, 1 mM) alone or in combination 

was assayed by 3 H-thymidine incorporation at different time 

points (12h, 24h and 48h). The effect of drug treatment in mitochondrial 

function was determined by monitoring changes in 

mitochondrial transmembrane potential (ΔΨm) and mitochondrial 

respiratory activity. ΔΨm was assessed by tetramethylrhodamine 

ethyl ester (TMRE)-labeling and analyzed by flow 

cytometry. Mitochondrial respiration (OXPHOS) and glycolysis 

were measured by real-time analysis of oxygen consumption 

(OCR) and extra-cellular acidification (ECAR) rates, respectively, 

in a XF96 Flux Analyzer. Expression of β-catenin and 

apoptosis-related proteins were analyzed by Western Blot. 

Results: FH535 and sorafenib inhibited HCC cell proliferation 

in all three different cell lines tested (Huh7, PLC and Hep3B). 

We observed a significant dose-response inhibition with single 

treatments, although the effect of combined drugs was 

more potent. Our results also indicated the decreased levels of 

ΔΨm in cells exposed to the simultaneous exposure to FH535 

and sorafenib, which may reflect the efficient mitochondrial 

drug-targeting of the combined treatment. Mitochondrial dysfunction 

was confirmed by lower oxygen consumption rates 

and increased levels of apoptosis in cells treated with FH535 

plus sorafenib. Similar ECAR levels in treated and non-treated 

cells ruled out a broad cellular metabolic failure and rather 

confirmed the predominant mitochondrial effect of the drugs. 

Conclusions: The combination of FH535 and sorafenib inhibits 

HCC cell proliferation. Mechanistically, our results suggest that 

the combined therapeutic targeting of β-catenin and RAS/RAF/ 

MAPK pathways may alter tumor cell growth by promoting 

mitochondrial dysfunction. 

Disclosures: 

The following authors have nothing to disclose: Lilia Turcios, Valery Vilchez, 

David Butterfield, Mihail I. Mitov, Francesc Marti, Roberto Gedaly 

674 

RA and butyrate induce liver and colon cancer cell 

apoptosis through miR-22 silencing of histone deacetylases 

Ying Hu, Hui-Xin Liu, Yu-Jui Yvonne Wan; Department of Medical 

Pathology and Laboratory Medicine, University of California, 

Davis Health Systems, Sacramento, CA 

Due to the signicant tumor suppressive role of microRNA-22 

(miR-22), it is important to establish the mechanism by which 

miR-22 expression is regulated. Our published data showed 

that bile acid-activated farnesoid x receptor (FXR) induces miR- 

22. In this report, we showed that all-trans retinoic acid (RA) 

and butyrate, normally present in digestive tract, also up-regulate 

miR-22. RA is a biologically active metabolite of vitamin 

A and a natural agonist for tumor suppressor retinoic acid 

receptor β (RARβ). Butyrate is a short chain fatty acid and 

histone deacetylase inhibitor (HDACi), produced by fiber-fed 

Gram-positive bacteria. Both agents can induce apoptosis in 

cancer cells. We tested the hypothesis that RA and butyrate 

act through miR-22 to inhibit HDACs and induce apoptosis in 

liver and colon cancer cells. Our data showed that RA/butyrate 

treatment synergistically promoted apoptosis and potently 

induced RARβ expression compared to single compound treatment 

in liver Huh7 and colon HCT116 cancer cells. miR-22 

induction by RA and butyrate was transcriptionally regulated 

by RARβ/RXRα and FXR/RXRα direct binding to a DR-5 motif 

located at -1568 to -1585 bp upstream of miR-22 based on 

transient transfection assays. Using 3’UTR luciferase assay, 

histone deacetylase 1 (HDAC1) was uncovered as a novel 

miR-22 target. Furthermore, miR-22 mimics also reduced the 

protein levels of HDAC4 and SIRT1, which are identified miR- 

22 targets in rat cardiomyocytes. Consistently, RA and butyrate 

reduced HDAC1, HDAC4, and SIRT1 protein levels in HCT116 

cells. In addition, the reduction in protein deacetylases was 

accompanied by increased histone acetylation in the RARβ 

regulatory region harboring a RA response element (DR-5). 

Histone modification of the RARβ gene could account for 

enhanced RARβ expression by RA/butyrate treatment. Moreover, 

miR-22 inhibitors abolished reduction of protein deacetylase 

levels and prevented apoptosis in HCT116 and Huh7 cells 

induced by RA/butyrate. Conclusion: RA and butyrate induce 

miR-22 to promote apoptosis of gastrointestinal cancer cells. 

miR-22 inhibition of HDACs is a novel pathway to explain the 

anti-carcinogenic effect of RARβ and FXR. 

Disclosures: 

The following authors have nothing to disclose: Ying Hu, Hui-Xin Liu, Yu-Jui 

Yvonne Wan 

675 

IRAKM-Mincle axis contributes to alcohol-induced liver 

injury via inflammasome activation 

Hao Zhou 1 , Laura E. Nagy 2 , Xiaoxia Li 1 ; 1 Immunology, Lerner 

Research Institute, Cleveland Clinic Foundation, Cleveland, OH; 

2 Pathobiology, Lerner Research Institute, Cleveland Clinic Foundation, 

Cleveland, OH 

Alcohol-induced liver injury is driven by hepatocyte necrosis 

and increased endotoxin translocation into the hepatic portal 

system, which together trigger chronic inflammatory liver 

damage. In this study, we found that mice deficient in IRAKM, 

a proximal Toll-like receptor pathway molecule, are protected 

from alcohol-induced liver injury. In response to low doses 

of LPS, which better reflect physiologically relevant levels of 

endotoxemia, IRAKM mediates the up-regulation of Mincle, a 

receptor for danger signals released by damaged cells. Biochemical 

analysis revealed that low-dose LPS preferentially 

induces the formation of an IRAKM Myddosome, which in turn 

promotes MEKK3-dependent NFκB activation. Mincle-deficient 

mice are also protected from alcohol-induced liver injury. Ex 

vivo studies demonstrated that both IRAKM and Mincle are 

required for inflammasome activation by the endogenous Mincle 

ligand SAP130, which is a danger signal release by damaged 

hepatocytes. Taken together, this study reveals a novel 

IRAKM-Mincle axis that critically mediates the pathogenesis of 

alcohol-induced liver disease through inflammasome activation. 

Disclosures: 

The following authors have nothing to disclose: Hao Zhou, Laura E. Nagy, Xiaoxia 

Li


676 

Hepatic maturation of induced Pluripotent Stem Cells 

is regulated by paracrine signals from endothelial and 

mesenchymal stem cells in culture and during organoid 

formation 

Akihiro Asai 1 , Eitaro Aihara 2 , Tatsuki Mizuochi 1 , Kieran Phelan 1 , 

Christopher Mayhew 1 , Pranavkumar Shivakumar 1 , Takanori 

Takebe 3 , James Wells 1 , Jorge A. Bezerra 1 ; 1 Pediatrics, Cincinnati 

Children’s Hospital Medical Center, Cincinnati, OH; 2 Department 

of Biology, University of Cincinnati, Cincinnati, OH; 3 Department 

of Regenerative Medicine, Yokohama City University, Yokohama, 

Japan 

Background: To recapitulate key stages of organ development, 

we engineered liver organoids using human induced pluripotent 

stem cells (iPSC), human mesenchymal stem cells (MSC), and 

human umbilical vein endothelial cells (HUVEC) as described 

previously. Because regulatory mechanisms of organoid maturation 

are not well defined, we investigated the differentiation 

patterns of liver organoids and tested if differentiation of iPSC 

requires direct contact with MSC and HUVEC. Methods: iPSC 

were differentiated to hepatic endoderm (iPS-HE) by Act,Wnt3a,DMSO,and 

Knock-Out Serum Replacement then co-cultured 

with MSC and HUVEC to generate 3D organoids (liver 

buds). Following implantation under kidney capsules of mice, 

plasma levels of human albumin (ALB) and alpha1-antitrypsin 

(A1AT) were monitored by ELISA. To investigate whether cellcell 

contact is a key mechanism by which MSC and HUVEC 

induce bud formation and maturation of iPS-hepatocytes, 

we cultured iPS-HE in the upper well of a transwell system, 

where MSC or HUVEC resided in the lower well individually 

or in co-culture. The expression of alphafetoprotein (AFP), 

HNF4a,ALB,A1AT,CPS1,BSEP,and CD31 was determined by 

immunostaining or by ELISA. Results: Temporal-spatial analysis 

by time-lapse microscopy revealed self assembly of iPS-HE, 

MSC, and HUVEC into a 3D tissue in 24 hr following a condensation 

and folding pattern to form a concave hemisphere. 

Hepatocytic differentiation in the buds was demonstrated by 

the AFP+/HNF4+ cells by whole-mount staining, in proximity to 

CD31+ endothelial cells. After implantation of the buds, human 

ALB (140-260ng/ml) and A1AT (62-174ng/ml) were detected 

in mouse plasma. Explanted organoids showed hepatocyte-like 

morphology organized in cellular clusters surrounded by vascular 

networks. Using the transwell culture to determine the role 

of paracrine effect, we found that ALB concentration in culture 

supernatants achieved 1587±233ng/ml/day in iPS-HE with 

MSC in the lower well (iPS/MSC) and 1881±162ng/ml/day 

in iPS-HE/HUVEC, which were higher than iPS-HE cultured with 

MSC+HUVEC in the lower (455±110ng/ml/day) or iPS-HE 

alone (81±21ng/ml/day, P


678 

Notch and Wnt control ductular reactions via the Igf1 

axis 

Sarah E. Minnis-Lyons 1 , Luke G. Boulter 2 , Tak Yung Man 1 , Michael 

J. Williams 1 , Rachel V. Guest 1 , Wei-Yu Lu 1 , Noemi Van Hul 4 , Isabelle 

A. Leclercq 4 , Owen J. Sansom 3 , Stuart J. Forbes 1 ; 1 MRC Centre 

for Regenerative Medicine, Edinburgh, United Kingdom; 2 MRC 

Human Genetics Unit, Edinburgh, United Kingdom; 3 Beatson Institute 

for Cancer Research, Glasgow, United Kingdom; 4 Universite 

Catholique de Louvain, Brussels, Belgium 

Ductular reactions (DR) are seen in chronic liver injury when 

hepatocyte regeneration is impaired and are thought to contain 

putative hepatic progenitor cells (HPCs). Their ability to contribute 

to large-scale parenchymal repair has been questioned. 

Notch and Wnt are key signals required for liver development 

and regeneration. Given their many actions include promoting 

cell expansion, we sought to identify if these signals control 

DRs after hepatocyte injury and importantly whether these 

pathways can be manipulated for future therapeutic targeting. 

Notch and Wnt pathways were analysed using a genetic in 

vivo model of hepatocellular injury and ductular activation that 

has allowed us to examine the temporal dynamics of the ductular 

response (AhCre MDM2fl/fl). We have also used K19 

and OPNCre lineage tracing tools, as well as HPC lines, to test 

small molecules, blocking antibodies and genetic loss of function 

in order to establish functionality and interactions of these 

signalling pathways. We have found distinct time-dependent 

Notch and Wnt signatures following hepatocyte injury. Small 

molecule inhibitors have revealed time-sensitive windows; with 

the early ductular proliferative response principally driven by 

Notch and the later response driven by Wnt. Anti-Notch1 

and Notch2 blocking antibodies and genetic loss of Notch3 

have confirmed DRs are driven by Notch1 and Notch3 but 

not Notch2. We have identified DRs as an additional source 

of the potent growth hormone Igf1 and this production is Wnt 

driven. Notch driven expression of Igf1-receptor within DRs 

highlights this axis as a node for cooperation between Notch 

and Wnt signals. Small molecule (AG1024) and genetic loss 

of function (Igf1 receptor fl/fl) confirms functionality of this axis 

and stresses a pivotal role for Igf1-receptor in the generation of 

the ductular reaction which can be enhanced by administration 

of recombinant Igf-1 (all P values =


However, the potential of hepatocytes to stably and functionally 

contribute to the biliary system is uncertain. Here, we use a 

mouse model of Alagille syndrome (ALGS) that lacks peripheral 

intrahepatic bile ducts (pIHBDs) to determine whether hepatocytes 

can transdifferentiate into normal cholangiocytes and 

assemble into functional bile ducts. In our ALGS mouse model 

genes encoding the NOTCH DNA binding partner RBP and the 

biliary transcription factor HNF6 are inactivated in embryonic 

liver progenitors by Cre-mediated recombination. Like humans 

with severe ALGS these mice lack pIHBDs at birth; remarkably, 

however, pIHBDs are formed in our ALGS mouse model with 

age. To determine whether hepatocytes are the source of the 

new pIHBDs in our Cre-based mouse model, we developed a 

Flp recombinase-mediated method of hepatocyte fate tracing. 

By following fate-traced hepatocytes through de novo pIHBD 

formation we determined that hepatocytes convert into cholangiocytes 

and undergo tubulogenesis. This results in a fully functional 

hepatocyte-derived biliary system capable of reverting 

cholestasis and liver injury. We found little clonal expansion of 

hepatocyte-derived cholangiocytes, which suggests that these 

cells do not transition through a transit amplifying cell stage, 

but rather many hepatocytes transdifferentiate and are incorporated 

into the new pIHBDs. Interestingly, we observed that 

biliary markers are activated in the hepatocytes near hypoxic 

regions, suggesting that local hypoxia may induce hepatocyte 

bile duct formation. Indeed, we show that activation of hypoxia 

signaling in hepatocytes in vivo can induce biliary differentiation. 

Our results demonstrate that hepatocytes can generate 

an intrahepatic biliary system effective in relieving liver injury 

in a mouse model of severe ALGS. These results define the 

plasticity of hepatocytes and add another dimension to the 

remodeling capacity of the adult liver. In addition, hepatocyte 

transdifferentiation independent of NOTCH signaling in our 

ALGS model suggests the existence of alternative drivers of 

biliary differentiation, like hypoxia signaling. Bile duct formation 

by hepatocytes may explain spontaneous improvement of 

cholestasis observed in some ALGS patients and could potentially 

be developed into an in vivo-reprogramming-based therapy 

for others. 

Disclosures: 

Johanna R. Schaub - Management Position: MDsave 

The following authors have nothing to disclose: Kari A. Huppert, Ashley E. Cast, 

Feng Chen, Stacey S. Huppert, Holger Willenbring 

681 

Identification of hepatic stem cell niche by label retaining 

cell assay with fetal and neonatal mice 

Reiichiro Kuwahara 1 , Neil D. Theise 2 , Takuji Torimura 1 ; 1 Department 

of Medicine, Division of Gastroenterology, Kurume University 

School of Medicine, Kurume, Japan; 2 Department of Pathology 

and Medicine, Beth Israel Medical Center of Albert Einstein College 

of Medicine, New York, NY 

Background and Aim: Previously we showed, in an acetaminophen 

injury model, that oval cells (OVc) appear to derive from 

the most proximal portions of the biliary tree (canals of Hering: 

CoH) in a time- and dose-dependent manner, and regenerating, 

differentiated peribiliary hepatocytes (PbH) also contributed 

to parenchymal repair (Kofman et al, Hepatology 2005; 

41: 1252). Subsequent label retaining cell (LRC) assays supported 

these findings (Kuwahara et al, Hepatology 2008; 47: 

1994). We have now investigated whether LRC assays could 

shed light on the possible role of CoH and PbH in normal liver 

development, in the absence of injury. Methods: Pregnant mice 

received water containing BrdU for scheduled term of 3 days 

(E10-12, E13-15, and E16-19). For the confirmation of BrdU 

labeling in the fetal liver, some mice were sacrificed immediately 

after each labeling term. The other mice were allowed to 

give birth to mice. Neonatal mice were raised to 8 weeks old 

to “chase” for label washout and then sacrificed. Other mice 

received intraperitoneal injection of BrdU for labeling at the 

age of 6, 13, 20, 27, 34, 41, 48 and 55 days. Then, some 

of them were sacrificed at 24 hours after each injection for the 

analyses of cell division. Other mice were raised to 8 weeks 

old to “chase” for label washout following the BrdU labeling 

and then sacrificed. PbH and biliary cell in CoH were analyzed 

by double immunostaining for biliary keratins (PanK)/ 

BrdU (cell division), PanK/Ki-67 (proliferation). Results: Almost 

all cells in the fetal liver were labelled with maternally administered 

BrdU in drinking water. In all mice with BrdU labeling 

during pregnancy, neither label retaining PbH nor label retaining 

biliary cells in CoH were detected in the liver of 8 weeks. 

The Ki67 index of neonatal mice liver showed that frequencies 

of cell proliferation of hepatocytes and biliary cells in CoH 

were highest at the age of 7 days and then diminished steadily. 

At 8 weeks post “chase”, distributions of BrdU-retaining PbH 

and biliary cells in CoH to total BrdU positive cells (lobular and 

peribiliary hepatocytes + biliary cells in CoH) were highest in 

the liver with BrdU labeling at the age of 13 days. These distributions 

were significant higher than those of liver with labeling 

at the age of 6, 20 and 27 days, respectively. Conclusion: 

These data provides support that PbH and CoH cells serve resident 

stem cell functions including contributing to parenchymal 

mass during organogenesis and early post-natal development 

(< 3weeks), but not after. 

Disclosures: 

The following authors have nothing to disclose: Reiichiro Kuwahara, Neil D. 

Theise, Takuji Torimura 

682 

Disruption of TGF-β-regulated CTCF Suppression of 

Telomerase links a Human Stem Cell Disorder to Liver 

Tumorigenesis 

Jian Chen 1 , Jiun-Sheng Chen 1 , Young Jin Gi 1 , H. Franklin Herlong 1 , 

Yun Seong Jeong 1 , Nipun Mistry 1 , Xiaoping Su 1 , Asif Rashid 1 , 

Bibhuti Mishra 1 , Jon White 2 , Milind Javle 1 , Marta L. Davila 1 , John 

R. Stroehlein 1 , Rosanna Weksbergc 3 , Jerry W. Shay, 4 , Keigo 

Machida 5 , Hidekazu Tsukamoto 5 , Lopa Mishra 1 ; 1 The University 

of Texas MD Anderson Cancer Center, Houston, TX; 2 Institute of 

Clinical Research, Veterans Affairs Medical Center, Washington 

DC, DC; 3 Hospital for Sick Children, Toronto, ON, Canada; 4 The 

University of Texas Southwestern Medical Center, Dallas, TX; 5 University 

of Southern California, Los Angeles, CA 

Patients with a human stem cell disorder, the Beckwith-Wiedemann 

syndrome (BWS) are known to develop multiple liver 

tumor types (hepatoblastoma, hepatocellular cancer, and cholangiocarcinoma) 

within a single patient. These patients are at 

an 800 fold increased risk of tumorigenesis. However, a precise 

mechanism for the switch in tumorigenesis remains elusive. 

In a previous study we demonstrated that loss of Transforming 

Growth Factor-beta (TGF-β) signaling, was a causal factor in 

BWS. To determine the mechanism for the oncogenic switch, 

we performed a broad bioinformatics and functional analyses 

utilizing human BWS cells, tissues and our mutant models in the 

TGF-β pathway. Methods and Results: (1) Over 80% of TGF-β/ 

Sptbn1 +/− /Smad3 +/− mutant mice spontaneously developed 

multiple tumors including the liver tumors that were phenotypically 

similar to those of patients with BWS. (2) Somatic mutations 

in SPTBN1 and SMAD3 are among the most frequent in 

human HCCs. (3) Both Sptbn1 +/− /Smad3 +/− mice and BWS 

cells express increased levels of stem cell-associated genes,


including Nanog, Sox2 and Oct4. (4) Increased levels of several 

stem-ness genes and telomerase (TERT) (25-45 fold), c-Myc 

(20-100 fold), IGF2 (~20 fold), with decreased levels of molecules 

implicated in BWS- P57 and CTCF were observed in liver 

tumors that developed in Sptbn1 +/- /Smad3 +/- mice. (5) Disruption 

of the β2SP/SMAD3/CTCF complex raises TERT levels 

in BWS and in Sptbn1 +/− /Smad3 +/− mice. (6) We observed 

that disruption of the β2SP/SMAD3/CTCF complex increases 

stem-like properties (increased ALDH-positive population and 

Sphere formation) and enhances tumorigenesis in human HCC 

cell lines. (7) We identified a mechanism by which the long 

range chromatin modulator CTCF facilitates TGF-β-mediated 

repression of hTERT transcription via β2S/SMAD3/CTCF interactions. 

Conclusions: We present new insights into liver tumor 

formation through the human stem cell disorder BWS and our 

mutant models in the TGF-β pathway, demonstrating a potential 

shift arising from disruption of TGF-β/CTCF signaling leading 

to BWS-associated tumorigenesis, telomerase activation, 

and human stem cell associated cancer development. This syndrome 

can provide new strategies for preventing and targeting 

liver cancers. 

Disclosures: 

The following authors have nothing to disclose: Jian Chen, Jiun-Sheng Chen, 

Young Jin Gi, H. Franklin Herlong, Yun Seong Jeong, Nipun Mistry, Xiaoping 

Su, Asif Rashid, Bibhuti Mishra, Jon White, Milind Javle, Marta L. Davila, John 

R. Stroehlein, Rosanna Weksbergc, Jerry W. Shay,, Keigo Machida, Hidekazu 

Tsukamoto, Lopa Mishra 

683 

Self-renewing diploid Axin2+ cells fuel homeostatic 

renewal of the liver 

Bruce M. Wang 1 , Roel Nusse 2,3 ; 1 Medicine, UCSF Liver Center, 

San Francisco, CA; 2 Developmental Biology, Stanford University, 

Stanford, CA; 3 Howard Hughes Medical Institute, Stanford, CA 

The cellular source of new hepatocytes in the adult liver and 

the molecular regulation of hepatocyte renewal are fundamental 

unanswered questions in liver biology. While it has been 

shown that new hepatocytes in the uninjured liver arise from 

pre-existing hepatocytes, hepatocytes are known to be heterogenous 

with striking differences in age and function across 

the liver lobule. It is unknown whether a specific subpopulation 

of hepatocytes serves homeostatic renewal in the liver. Using 

a novel mouse model for stably labeling Wnt-responsive cells 

in vivo, we have discovered a unique population of Wnt-controlled 

hepatocytes with stem cell characteristics surrounding 

the central vein. These pericentral cells express the early liver 

progenitor marker Tbx3, proliferate at a faster rate than other 

hepatocytes and are diploid, and thus differ from mature 

hepatocytes, which are mostly polyploid. Over time, these 

cells self-renew and give rise to descendants that differentiate 

into Tbx3-negative, polyploid hepatocytes and can replace all 

hepatocytes along the liver lobule during homeostatic renewal. 

Importantly, these Wnt-responsive cells are present in the normal 

liver, thereby distinguishing them from injury-induced multipotent 

progenitor cells (oval cells) associated with the portal 

region. Adjacent central vein endothelial cells provide essential 

Wnt signals that maintain the pericentral cell population, 

thereby constituting the hepatocyte stem cell niche. Thus, we 

describe for the first time a subset of hepatocytes that subserves 

homeostatic hepatocyte renewal, characterize its anatomical 

niche, and identify molecular signals that regulate its activity. 

Disclosures: 

The following authors have nothing to disclose: Bruce M. Wang, Roel Nusse 

684 

FAP disease modelling using patient-specific iPS cells 

derived from urine 

Christoph J. Niemietz, Vanessa Sauer, Jacquelline Stella, Gursimran 

Chandhok, Andree Zibert, Hartmut H. Schmidt; Klinik für 

Transplantationsmedizin, Universitätsklinikum Münster, Münster, 

Germany 

Transthyretin-related familial amyloid polyneuropathy (TTR- 

FAP) is a rare genetic neurodegenerative disease caused by 

mutations of TTR. TTR is primarily secreted by the liver and 

misfolding ultimately leads to aggregation and fibril deposition 

in peripheral tissues and organs. In order to prevent expression 

of TTR, gene silencing strategies are currently under clinical 

investigation. The use of primary cells derived from FAP 

patients for evaluation of antisense strategies is difficult. As 

an alternate, stem cell technology is used here to generate 

patient specific primary cells. Urine collections of FAP patients 

were processed for isolation of renal epithelial cells, followed 

by reprogramming into iPS cells (iPSCs) using non-integrating 

episomal vectors. After characterization of pluripotent cell lines, 

differentiation towards hepatocyte-like cells was accomplished 

after treatment with factors for 14 days. iPSC-Heps were characterized 

by analysis of typical hepatic markers via qRT-PCR, 

flow cytometry, and immunocytochemistry. ASOs and siRNAs 

were introduced into iPSC-Heps in order to evaluate gene 

silencing of TTR via qRT-PCR analysis, ELISA and Western-blot. 

Typically, 2-3 stable cell colonies emerged upon cultivation 

of urine-derived cells from FAP patients (n=12). iPSC-Heps 

showed similar morphology and gene expression in comparison 

to HepG2 control. TTR gene expression was high. ASOs or 

siRNA treatment resulted in downregulation of TTR. Our results 

indicate that iPSCs derived from urine cells of FAP patients can 

be routinely reprogrammed and differentiated into iPSC-Heps 

expressing TTR at comparable levels as control cells. TTR gene 

silencing analysis of iPSC-Heps allows the establishment of a 

patient-specific in vitro platform for evaluating drug efficiency.


Thus, the technology is excellently suited for modelling of liver 

disease and may allow assessment of novel drugs for improved 

therapy of FAP. 

Disclosures: 

The following authors have nothing to disclose: Christoph J. Niemietz, Vanessa 

Sauer, Jacquelline Stella, Gursimran Chandhok, Andree Zibert, Hartmut H. 

Schmidt 

685 

CD34+ Liver Cancer Stem Cells were Formed by Fusion 

of Hepatobiliary Stem/Progenitor Cells with Hematopoietic 

Precursor-Derived Myeloid Intermediates 

Changjun Zeng 2 , Yanling Zhang 2 , Su Cheol Park 2 , Jong Ryeol 

Eun 2 , Ngoc Tue X. Nguyen 2 , Benjamin Tschudy-Seney 2 , Yong 

Jin Jung 2 , Neil D. Theise 3 , Mark Zern 2 , Yuyou Duan 1 ; 1 Dermatology, 

Internal Medicine, Institute for Regenerative Cures, UC Davis 

Medical Center, Sacramento, CA; 2 Internal Medicine, Institute for 

Regenerative Cures, University of California Davis Medical Center, 

Sacramento, CA; 3 Beth Israel Medical Center, Albert Einstein College 

of Medicine, New York, NY 

A large number of cancer stem cells (CSC) were identified and 

characterized; however, the origins and formation of these 

CSC remain elusive. Here, we examined the origination of 

newly-identified CD34+ liver CSC (LCSC). CD34+ LCSC was 

clonogenically cultured in vitro employing our newly-developed 

medium, and cloned CD34+ LCSC was used to inject 

into NOD/SCID mice to generate the xenografts of human 

liver carcinomas (HLC). Our results that CD34+ LCSC co-expressed 

liver stem cell and myelomonocytic cell markers, showing 

a mixed phenotype of hepatobiliary stem/progenitor cells 

(HSPC) and myelomonocytic cells, as determined by both qPCR 

and Immunohistochemistry. Moreover, human xenografts and 

parental cells which CD34+ LCSC was isolated, co-expressed 

liver cancer and meylomonocytic markers, also demonstrating 

mixed phenotypes. ELISA assay showed that the xenografts 

and parental cells secreted albumin demonstrating its hepatocyte 

origin, and these cells also expressed cytokines (IL-1b, IL-6, 

IL-12A, IL-18, TNF-α, and CSF1) and chemokines (IL-8, CCL2, 

and CCL5), it is well known that the macrophage is the primary 

cell which produces these factors. Expression of these cytokines 

and chemokines responded to the stimuli (INF-γ, IL-4, and LPS), 

response to stimulation by these three factors is a unique macrophage 

characteristic. Furthermore, human xenografts and 

the parental cells phagocytized E. Coli; thus, they appeared to 

express characteristics of the two parental cell types as hybrid 

cells do. CD34+ LCSC co-expressed CD45, demonstrating its 

origin appears to be from a hematopoietic precursor. The percentage 

of cells positive for OV6, CD34, and CD31 presenting 

the markers of HSPC, hematopoietic and myelomonocytic 

cells, were increased under treatment of CD34+ LCSC with 

drug cisplatin. Cytogenetic analysis showed that CD34+ LCSC 

contained a greater number of chromosomes (aneuploid). 

HBV DNA integrations and mutations in CD34+ LCSC and the 

parental cells, were identical to those in the literature published 

three decades ago or those in the Sanger COSMIC database. 

In conclusion, these results demonstrated that CD34+ LCSC 

were formed by fusion of HSPC with CD34+ hematopoietic 

precursor-derived myeloid intermediates, thus, we believe that 

this is the first report that human CSC appears to have been 

formed by the fusion, and it is also the first report that a HLC 

appears to be initiated and developed from the interaction of 

HSPCs with CD34+ hematopoietic precursors, thus revealing a 

diversity of origins for HLC. Therefore, our results represent a 

significant step towards better understanding of the formation 

of human CSC, and the diverse origins of human cancers. 

Disclosures: 

The following authors have nothing to disclose: Changjun Zeng, Yanling Zhang, 

Su Cheol Park, Jong Ryeol Eun, Ngoc Tue X. Nguyen, Benjamin Tschudy-Seney, 

Yong Jin Jung, Neil D. Theise, Mark Zern, Yuyou Duan 

686 

TGF-β inhibits lncRNA H19 expression via SOX2 in 

tumor-initiating hepatocytes 

Jinqiang Zhang, Chang Han, Weina Chen, Kyoungsub Song, Ying 

Wang, Lu Yao, Nathan Ungerleider, Hyunjoo Kwon, Tong Wu; 

Pathology and Laboratory Medicine, Tulane University, New Orleans, 

LA 

TGFβ pathway modulates various biological processes. The 

output of a TGFβ response is highly depending on tissue types 

and diseases. This is especially true in hepatocellular carcinoma 

(HCC) initiation and progression, where the tumor-initialing 

and malignant cells are highly influenced by the liver 

environments. To delineate the role of TGFβ in the process 

of hepatocarcinogenesis, we employed a novel tumor-initiating 

cell transplantation system that avoids the possible effects 

incurred from genetic manipulation of background liver. Specifically, 

TGFβ receptor II (Tgfbr2) flox/flox mice at 14 days of 

age were given one dose (25mg/g) of the hepatic carcinogen 

diethylnitrosamine via intraperitoneal injection. Three months 

later, the initiated hepatocytes from Tgfbr2 flox/flox mice 

were isolated and transplanted to the same strain (C57Bl/6) 

wild type recipient mice. Four weeks later, the recipient mice 

received one does of Cre recombinase adenovirus (Ad-Cre) 

through tail vein to delete Tgfbr2 in the transplanted tumor-initiating 

hepatocytes; separate group of the recipient mice were 

injected with the control adenovirus (Ad-GFP). The recipient 

mice were closely monitored for liver tumor burden. We 

observed that deletion of Tgfbr2 in tumor-initiating hepatocytes 

by tail vein injection of Ad-Cre led to formation of more and 

bigger liver tumors (compared to Ad-GFP group). This finding 

suggests that TGFβ inhibits the malignant potential of tumor-initiating 

hepatocytes. In parallel, we further analyzed the effect 

of TGFβ in tumor initiating hepatocytes in vitro. Specifically, 

tumor-initiating hepatocytes isolated from DEN-treated Tgfbr2 

flox/flox mice were infected with Ad-Cre or Ad-GFP prior to 

TGFβ1 treatment in vitro; RNAs were then isolated for transcriptome 

sequencing analysis. We identified a group of lncRNAs 

that were noticeably regulated by TGFβ, including the lncRNA 

H19. We found that deletion of Tgfbr2 by Ad-Cre in tumor initiating 

hepatocytes led to a 5-fold increase of H19 expression. 

We observed that deletion of H19 by siRNA decreased the 

tumor-initiating cell property, whereas forced overexpression 

of H19 enhanced it. Chromatin immunoprecipitation assay 

showed that SOX2 bound to the promoter region of H19 gene. 

While SOX2 overexpression in tumor-initiating hepatocytes 

enhanced H19 transcription, SOX2 knockdown reduced it. 

Furthermore, TGFβ treatment reduced the protein level of SOX2 

in tumor-initiating hepatocytes. Taken together, our findings 

disclose a novel mechanism that importantly regulates hepatocarcinogenesis 

—- TGFβ inhibits H19 expression via SOX2 in 

tumor-initiating hepatocytes and this effect leads to inhibition of 

HCC development. 

Disclosures: 

The following authors have nothing to disclose: Jinqiang Zhang, Chang Han, 

Weina Chen, Kyoungsub Song, Ying Wang, Lu Yao, Nathan Ungerleider, Hyunjoo 

Kwon, Tong Wu


687 

Expression pattern of the stem/progenitor cell marker 

Neighbor of Punc E 11 in different mouse models of 

liver regeneration. 

Vera Hoffmann, Andrea Bowe, Harald M. Curth, Tobias Goeser, 

Dirk Nierhoff; University Hospital of Cologne, Department of Gastroenterology 

and Hepatology, Cologne, Germany 

Background: Regeneration after acute liver injury naturally 

arises from the compartment of parenchymal liver cells. However, 

if proliferation capacity of hepatocytes is inadequate, 

liver progenitor cells are activated and can give rise to mature 

hepatocytes and cholangiocytes. In this project, we have 

focused on the expression pattern of the novel stem/progenitor 

cell marker Neighbor of Punc E 11 (Nope) in liver regeneration 

after acute and chronic liver injury. Methods: C57Bl6 mice on 

normal chow, on a 3,5-diethoxycarbonyl-1,4-dihydro-collidin 

(DDC) or a choline-deficient, ethionine-supplemented (CDE) 

diet were investigated. DDC and CDE diet were optionally 

followed by a regenerative period on normal chow. For additional 

acute liver injury, subgroups of normal mice as well as 

mice on a DDC diet were subjected to a partial hepatectomy 

(PH) with or without retrorsine to block hepatocyte proliferation. 

After defined time periods between 3 to 6 weeks, mice were 

sacrificed and quantitative RT-PCR and immunohistochemical 

costainings of the liver tissue for Nope with CK19, A6, EpCAM 

(oval cell markers), E-cadherin or HNF4α (hepatocyte markers) 

were performed. Results: Partial hepatectomy with or without 

retrorsine did not induce any expression of Nope above background 

level in the adult liver. After DDC diet the expression 

level of Nope was increased in the RT-PCR. In the CDE model, 

the expression level of Nope even reached the same level as 

in the fetal liver. We were able to detect ductular proliferations 

with coexpression of CK19, A6 and EpCAM with Nope. 

In the DDC model, only A6 also stained positive in a minor 

cell fraction within periductular Nope positive hepatocytic cell 

populations. After PH in the DDC model, followed by normal 

chow, the Nope expression level decreased, but small hepatocytic 

cells in proximity to ductular structures stained positiv for 

Nope. In the CDE model, we were able to detect clearly confined 

Nope positive hepatocytic cell clusters mainly in proximity 

to Nope positive ductular structures. Clearly distinguishable 

from these Nope expressing cell populations, expression of 

Nope was infrequently induced in HNF4α positive parenchymal 

hepatocytes. Discussion: The oncofetal marker Nope is 

expressed in CK19, A6 and EpCAM positive ductular cells as 

well as in CK19 negative periductular small hepatocytic cells 

presumably representing early hepatocytes after liver injury. 

These Nope positive early hepatocytes can form confined 

regenerative clusters with a minor fraction staining positive for 

A6. In conclusion, Nope is a marker for progenitor cells and 

periductular early hepatocytes in different mouse models of 

liver regeneration. 

Disclosures: 

Tobias Goeser - Advisory Committees or Review Panels: Gilead, Johnson, BMS, 

BMS; Grant/Research Support: Roche; Speaking and Teaching: Essex, BMS, 

Gilead, Novartis, Falk, Roche, Essex, BMS, Gilead, Novartis, Falk 

Dirk Nierhoff - Consulting: AbbVie; Speaking and Teaching: Gilead, AbbVie, 

BMS, Janssen, MSD 

The following authors have nothing to disclose: Vera Hoffmann, Andrea Bowe, 

Harald M. Curth 

688 

Ex vivo-expansion of human CD34 + cells from patients 

with liver cirrhosis enhances therapeutic efficacy of cell 

transplantation for rat cirrhotic liver 

Toru Nakamura 1,2 , Hironori Koga 1,2 , Hideki Iwamoto 1,2 , Yu 

Ikezono 1,2 , Fumitaka Wada 1,2 , Mitsuhiko Abe 1,2 , Takahiko 

Sakaue 1,2 , Takato Ueno 3,2 , Takuji Torimura 1 ; 1 Division of Gastroenterology, 

Department of Medicine, Kurume University School of 

Medicine, Kurume, Japan; 2 Liver Cancer Division, Research Center 

for Innovative Cancer Therapy, Kurume University, Kurume, Japan; 

3 Asakura Medical Association Hospital, Asakura, Japan 

Background: We demonstrated that the transplantation of 

human CD34 + cells into an immunodeficient rat liver fibrosis 

model reduced liver fibrosis by suppressing activated hepatic 

stellate cells and increasing MMPs activity, and led to hepatic 

regeneration. Recently, we reported that autologous granulocyte-colony 

stimulating factor (G-CSF)-mobilized CD34 + cell 

transplantation for patients with decompensated liver cirrhosis 

(LC) had therapeutic potential, but the colony-forming ability 

of CD34 + cells from patients with decompensated LC was 

reduced. Thus, recovery of CD34 + cell function is indispensable 

for cell transplantation therapy of patients with LC. The aim of 

this study was to investigate the efficacy of cell transplantation 

therapy with ex vivo-expanded human CD34 + cells for carbon 

tetrachloride (CCl 4 

)-induced liver fibrosis model. Methods: 

Human G-CSF-mobilized CD34 + cells of patients with LC were 

isolated by magnetic cell sorting system. Recipient nude rats 

were injected intraperitoneally with CCl 4 

twice weekly for 3 

weeks before initial treatment. Then, saline, 5×10 4 , or 1×10 6 

non-expanded and expanded CD34 + cells/kg body weight 

were transplanted via spleen, respectively. The administration 

of CCl 4 

was continued for three more weeks until the rats were 

sacrificed. Examination items were as follows: 1) FACS and 

Real-Time PCR analysis of non-expanded and expanded CD34 + 

cells, 2) morphometry of fibrotic areas by Azan-Mallory staining, 

3) immunohistochemistry using anti-CD31, smooth muscle 

myosin heavy chain-1 (SM1), αSMA, Ki67, and PCNA antibodies, 

and 4) the RT 2 Profiler TM PCR Array analysis. Results: 

For 7 days, CD34 + cells were effectively expanded. Increased 

expression of VE-cadherin, KDR and Tie-2 was determined by 

FACS analysis. The expression of pro-angiogenic growth factors 

in expanded CD34 + cells increased compared with non-expanded 

CD34 + cells. The transplanted cells differentiated into 

CD31 + and SM1 + cells. Expanded CD34 + cell transplantation 

had dose-dependently reduced liver fibrosis. Assessments 

of hepatocytes and sinusoidal endothelial cells proliferative 

activity indicated the superior potency of expanded CD34 + 

cells over non-expanded CD34 + cells. The PCR Array analysis 

against the adhesion molecules was showed that the expression 

of integrin αvβ3 was the most up-regulated gene compared 

with before culture. Three weeks of treatment with Cilengitide, 

which specially inhibits integrin αvβ3 signaling, resulted in the 

inhibition of CD34 + cells migration and worsened liver fibrosis 

in a dose-dependent manner. Conclusion: These findings suggest 

that expanded CD34 + cell transplantation promote better 

therapeutic effects for liver cirrhosis. 

Disclosures: 

The following authors have nothing to disclose: Toru Nakamura, Hironori Koga, 

Hideki Iwamoto, Yu Ikezono, Fumitaka Wada, Mitsuhiko Abe, Takahiko Sakaue, 

Takato Ueno, Takuji Torimura


689 

Attenuation of liver fibrosis development by angiotensin 

2-receptor blocker 1- treatment augments hepatocyte 

differentiation of hepatic progenitor cells 

Mitsuteru Kitade, Norihisa Nishimura, Hitoshi Yoshiji; Nara Medical 

University, Kashihara, Japan 

Recent research has elucidated mechanisms and potentials of 

hepatic progenitor cells (HPC) on regeneration of diseased 

liver. It is commonly known that hepatic stellate cells (HSC) 

play important roles as niche component cells of HPC, and 

that HPC expands along with HSC activation and liver fibrosis 

development. However, less is known about whether liver fibrosis 

development affects HPC-mediated regeneration. We have 

reported that angiotensin 2 strongly augments proliferation of 

activated-HSC, and angiotensin 2-receptor blocker 1 (ARB) 

exerts anti-fibrotic effect on diseased liver by inhibition of HSC 

proliferation. Our current study was performed to elucidate 

interactions between HSC and HPC-mediated liver regeneration 

using losartan, an ARB reagent on a DDC-induced mouse 

liver model. DDC-treatment augmented liver injury with fibrosis 

development and HPC expansion. ARB-treatment attenuated 

both a-SMA-positive activated HSC and laminin-positive extracellular 

matrix with increased hepatocyte (HC) differentiation 

of HPC, which may result in gain of liver/body weight ratio. 

Our in vitro experiments showed that ARB treatment did not 

alter capacity for HPC differentiation. In contrast, conditional 

medium of human HSC line (LX-2) augmented biliary epithelial 

cell (BEC) differentiation of HPC but attenuated efficiency for 

hepatocyte differentiation. Conditional medium of LX-2 incubated 

with angiotensin 2 enhanced this polarity of HPC differentiation 

towards BEC, which was blocked by ARB. These 

results indicated that HSC augments HPC differentiation fates 

towards BEC in paracrine manner. We also confirmed that 

both LX-2 and primary activated HSC strongly express Jagged1, 

a ligand for NOTCH, which plays central role in HPC 

differentiation fate towards BEC, which may controls HPC differentiation 

fate towards BEC. In conclusion, anti-fibrosis treatment 

may also be beneficial on efficient HPC-mediated liver 

regeneration via preferential differentiation of HPC towards HC 

by blocking NOTCH pathway in HPC niche. 

Disclosures: 

The following authors have nothing to disclose: Mitsuteru Kitade, Norihisa 

Nishimura, Hitoshi Yoshiji 

690 

Bone Marrow Mesenchymal Stem Cells (BMSCS) From 

Decompensated Cirrhotics Show Premature Senescence 

And Limited Tissue Repair And Regenerative Potential 

Dhananjay Kumar 1 , Sheetalnath B. Rooge 1 , Smriti Shubham 1 , 

Lovkesh Anand 2 , Sujata Mohanty 3 , Chhagan Bihari 4 , Anupam 

Kumar 1 , Shiv K. Sarin 2 ; 1 Department of Research, Institute of Liver 

and Biliary Sciences (ILBS), New Delhi, India; 2 Department of 

Hepatology, Institute of Liver and Biliary Sciences, New Delhi, 

India; 3 Stem Cell Facility, All India Institute of Medical Sciences, 

New Delhi, India; 4 Department of Pathology, Institute of Liver and 

Biliary Sciences, New Delhi, India 

Background & aim: Use of autologous BMSCs in chronic liver 

disease have shown varied clinical response ranging from negligible 

to mild improvement in CTP scores. Whether BMSCs 

from CLD patients are functionally efficient for repair & regeneration 

of cirrhotic liver is largely unknown. We investigated 

the functional potential of BMSCs of decompensated liver disease 

(DLD) patients. Patients & Methods: MSCs were isolated 

from DLD(N=10) patients & matched healthy BM donors(N=8) 

& characterized by surface marker expression & in-vitro differentiation 

to adipocytes & osteocytes. Levels of trophic & 

paracrine factors (RTPCR & cytokine array), angiogenic properties 

& immunomodulatory functions were studied. Population 

doubling time, CFU & SA-βGal staining were used to study 

cellular aging & senescence. Results: All MSCs fulfilled the minimal 

criteria for mesenchymal stromal cells; plastic adherence, 

spindle-shaped morphology, inducible osteo & adepogenesis 

& specific surface expression patterns. In RT-PCR analysis, 

DLD-MSCs(dMSCs) showed more than 2 fold decrease in 

expression of SDF1, HGF, FGF2, ANG1, JAG1 in comparison 

to healthy-MSCs(hMSCs). dMSCs also showed reduced capacity 

(p


bution of GFP was inversely correlated with alpha-fetoprotein 

(AFP) expression, suggesting local insulin signaling mediated 

by IRS2 may play a role in tumor heterogeneity and cellular 

differentiation. Exogenous expression of IRS2 promoted tumorigenesis 

in HepG2 soft agar and clonogenic assays and also 

corresponded with decreased AFP immunostaining. In HepaRG 

pIRS2-GFP positive cells were observed in discrete sites within 

the cultures surrounding “islands” of hepatocyte differentiation. 

IRS2 enriched cells had progenitor-like properties and we show 

that insulin/IRS2 signaling at a cellular level determines the 

differentiation, proliferative expansion and survival of hepatocyte-like 

tumor cells within the cultures, whilst expression of 

IRS2 is dynamically regulated during the timecourse of hepatocyte 

differentiation such that levels diminished as cells matured. 

Taken together, our results underscore the heterogeneity of 

insulin sensitivity at the cellular level within “homogeneous” 

cancer cell lines. We show that IRS2 expression is spatially 

patterned both in monolayers in vitro and in tumors in vivo and 

serves as a proxy for insulin sensitivity. Further investigation of 

how this novel cellular niche reacts to aberrant insulin signaling 

in the context of metabolic disease may provide future insights 

into the links that underpin the association between type II diabetes 

and HCC risk. 

Disclosures: 

The following authors have nothing to disclose: Fátima Manzano Núñez, Carlos 

Acosta Umanzor, Aránzazu Leal Tassias, Deborah J. Burks, Luke A. Noon 

692 

Sonic Hedgehog-Containing Exosomes Mediate Biliary 

Differentiation and Fibronectin Deposition via Epigenetic 

Regulation 

Nidhi Jalan-Sakrikar, Thiago de Assuncao, Jie Lu, Gwen Lomberk, 

Martin E. Fernandez-Zapico, Raul A. Urrutia, Robert C. Huebert; 

Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN 

Background: Developmental morphogens play an important 

role in coordinating the ductular reaction and portal fibrosis 

that occur in the setting of cholangiopathies. However, little is 

known about how injured cholangiocytes signal to adjacent 

progenitor cells to promote repair after biliary injury. Recent 

studies show that sonic hedgehog (Shh) signaling is activated 

during liver regeneration and repair. In addition, cholangiocytes 

release exosomes that contain biologically active Shh and 

the secretion of these organelles increases during biliary fibrosis. 

Our lab has recently reported a differentiation protocol that 

generates induced pluripotent stem cell (iPSC)-derived cholangiocytes 

(iDC), a model useful for studying the stem cell to cholangiocyte 

transition. The purpose of this study was to dissect 

the mechanisms whereby injured cholangiocytes communicate 

with the neighboring progenitor cells via Shh-containing exosomes 

to initiate both cholangiocyte differentiation and extracellular 

matrix (ECM) remodeling. Methods and Results: Next 

generation RNA sequencing at each phase of the iPSC to iDC 

transition revealed significant upregulation of the Smoothened 

(SMO) receptor as well as Gli-1 and Gli-2 transcription factors 

during biliary specification and these changes correlated 

with a 60-fold increase in fibronectin (FN) levels. Exosomes 

isolated from LPS-injured cholangiocytes enhanced progenitor 

cell acquisition of cholangiocyte markers (CK7: 2.7±0.3-fold; 

CK19: 2.3±0.2-fold) as well as the expression and release 

of FN (8.3±2.4 fold). Cholangiocyte differentiation and FN 

release were both mediated by Shh as shown using pharmacologic 

(cyclopamine) or genetic (SMO shRNA) inhibition of 

Shh signaling. Concurrent alterations in epigenetic regulators 

(SetD7, KDM5D, EZH2) suggested that a common epigenetic 

regulatory program, driven by Shh, may mediate both cholangiocyte 

differentiation and ECM deposition. Cholangiocyte 

differentiation was associated with temporal alterations in histone 

methylation patterns that were Shh-responsive, including 

early increases in H3K4me3 (4.6±1.6-fold) and late decreases 

in H3K27me3 (-3.8±1.2-fold). In vivo, mice fed the choline-deficient, 

ethanolamine supplemented diet had an expansion of 

LGR5+ progenitor cells that was associated with Shh activation, 

increased FN deposition, and peri-portal fibrosis. Conclusions: 

We conclude that Shh-containing exosomes play an 

integral role in cholangiocyte differentiation from progenitor 

cells following biliary injury and that an epigenetically-driven 

cascade of gene regulation may simultaneously promote FN 

deposition and biliary fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Nidhi Jalan-Sakrikar, Thiago de 

Assuncao, Jie Lu, Gwen Lomberk, Martin E. Fernandez-Zapico, Raul A. Urrutia, 

Robert C. Huebert 

693 

Interleukin-17 mediates liver progenitor cell transformation 

into cancer stem cells in hepatocellular carcinoma. 

Imène Gasmi 1,2 , Adrien Guillot 2,3 , Nabila Hamdaoui 1,2 , Arthur 

Brouillet 1,2 , Julien Calderaro 1,2 , Benoit Rousseau 1,2 , Jean-Michel 

Pawlotsky 1,2 , Fouad Lafdil 1,2 ; 1 INSERM U955 Henri Mondor Hospital, 

INSERM / University of Paris Est, Créteil, France; 2 INSERM 

/ University of Paris Est, Creteil, France; 3 NIH NIAAA, Bethesda, 

MD 

Introduction. Hepatocellular carcinoma (HCC) is the third leading 

cause of cancer-related death. HCC arises in the setting 

of cirrhotic livers in 80 to 90% of cases and progresses in an 

inflammatory context. Cancer stem cell biology has recently 

sparked the interest of scientists because of their capacity to 

initiate and to enhance tumor progression. However the underlying 

mechanisms by which expansion of CSCs is initiated are 

still unclear. After severe and chronic liver injury, liver progenitor 

cell compartment is activated under sustained inflammatory 

response. LPCs participate to the regenerative process and are 

also observed in HCC. Interestingly, among the large spectrum 

of released cytokines in HCC, recent studies identified 

IL-17-producing cells as a factor associated with a poor prognosis 

of the disease. In this study, we propose to determine 

whether IL-17 could be involved in tumorigenesis, in particular, 

by promoting the transformation of resident liver progenitor 

cells (LPCs) into CSCs. Materiels and methods. Serial sections 

from 70 HCC-patients were immuno-stained to identify LPCs 

(with anti-CK19) and IL-17-producing cells (with anti-IL-17). In 

vitro, a murine LPC line (BMOL) was used for long-term culture 

with or without IL-17 for 10, 20, 30 or 40 days. The expression 

of cancer stem cell markers were analyzed by quantitative 

RT-PCR and flow cytometry. Cell cycle controlling factors 

were assessed by western blot. Acquired self-renewal property 

of LPCs after long term culture with IL-17 was assessed by 

spheroid formation capacity analysis. Results. Identification 

and semi-quantitative analysis of CK19+ and IL-17+ cells in 

HCC patients showed a positive correlation between the number 

of infiltrated IL-17+ cells and the number of LPCs. In vitro, 

LPC stimulation with IL-17 led to increased expression of CSC 

marker mRNA expression including Klf4, ALDH1A1 or CD133, 

EpCAM and Glypican-3. CD133 protein and ALDH1A1 activity 

analyzed by flow cytometry, were enhanced in LPC cultured 

for 10, 20 or 30 days when compared to none treated LPCs. 

Cell cycle analysis showed that IL-17 induces the expression 

of the proto-oncogene c-Raf, and cyclin D1. In addition, LPCs 

cultured for 10 days in presence of IL-17 were able thereafter 

to form spheroids when transferred in low-attachment culture


dishes, in contrast to none pre-treated LPCs. Conclusion. Taken 

together, these results strongly suggest that IL-17 could contribute 

to HCC development, especially by leading LPC to acquire 

cancer stem cell-like properties. Therefore, IL-17 neutralization 

or IL-17 signaling pathway disruption in LPCs may contribute to 

CSC niche eradication in HCC treatment. 

Disclosures: 

Jean-Michel Pawlotsky - Consulting: Abbvie, Achillion, Bristol-Myers Squibb, Gilead, 

Janssen, Merck; Speaking and Teaching: Bristol-Myers Squibb, Gilead, 

Merck, Janssen 

The following authors have nothing to disclose: Imène Gasmi, Adrien Guillot, 

Nabila Hamdaoui, Arthur Brouillet, Julien Calderaro, Benoit Rousseau, Fouad 

Lafdil 

694 

Growth Factor Mobilized CD34+ Bone Marrow Stem 

Cells (BMSCs) Rescue the Loss of Regenerative Microenvironment 

in Decompensated liver Cirrhosis 

Sheetalnath B. Rooge 1 , Lovkesh Anand 2 , Dhananjay Kumar 1 , 

Smriti Shubham 1 , Rakhi Maiwall 2 , Chhagan Bihari 3 , Anupam 

Kumar 1 , Shiv K. Sarin 2 ; 1 Department of Research, Institute of 

Liver and Biliary Sciences (ILBS), New Delhi, India; 2 Department 

of Hepatology, Institute of Liver and Biliary Sciences (ILBS), New 

Delhi, India; 3 Department of Pathology, Institute of Liver and Biliary 

Sciences (ILBS), New Delhi, India 

Background: Therapeutic effect of growth factor mobilized 

CD34+ BMSCs in management of chronic Liver Disease (CLD) 

has been shown by several investigators including our group 

(Gastroenterology 2012, 2015). However the underlying 

mechanisms operating in the hepatic tissue are not clearly 

understood. Aim: To study the cellular & molecular mechanisms 

of growth factor activities on liver regeneration in CLD 

patients. Patients & Method: Fourty-one Patients with cirrhosis 

were administered growth factors [G-CSF at 5mcg/kg at days 

1,2,3,4,5 & then every 3rd day till day 60, Erythropoietin 

at 500 I.U/Kg s/c twice a week for 2 months & tranjugular 

liver biopsy & hepatic vein samples were obtained before 

& after the therapy. Regenerative response was studied by 

immunohistochemistry (IHC) using CK19 for hepatic progenitor 

cells, Ki67 for hepatocyte replication. Associated cellular 

microenvironment was analyzed by IHC by using cell type 

specific markers i.e. myofibroblasts (α-SMA), BMSCs (CD34), 

tissue macrophage (CD68) & M2 macrophages (CD163). 

Further, to study the change in secretary microenvironment a 

panel of cytokines, chemokines & growth factors were analyzed 

in hepatic vein plasma using Millipore Milliplex Map 

Kit & compared between responders (


Fig 1 

results demonstrated that TSG-6 promoted the autophagical 

function in hepatocytes, contributing to the liver regeneration. 

Disclosures: 

The following authors have nothing to disclose: Sihyung Wang, Jeongeun Hyun, 

Jieun Kim, Youngmi Jung 

Disclosures: 

The following authors have nothing to disclose: Ritu Khosla, Archana Rastogi, 

Shyam Singh, Madavan Vasudevan, Viniyendra Pamecha, Gayatri Ramakrishna, 

Shiv K. Sarin, Nirupma Trehanpati 

696 

TSG-6 enhances autophagy in chronically damaged 

liver 

Sihyung Wang, Jeongeun Hyun, Jieun Kim, Youngmi Jung; Pusan 

National University, Pusan, Korea (the Republic of) 

Tumor necrosis factor-inducible gene 6 protein (TSG-6), one 

of cytokines secreted from mesenchymal stem/stromal cells, is 

known to act as an anti-inflammatory factor and reduce several 

pathological conditions. Recently, TSG-6 has been shown to 

protect liver from injury; however, the mechanism underlying the 

effect is poorly understood. Autophagy is the catabolic process 

that targets cell constituents to the lysosomes for degradation 

and known to be dysregulated in several diseases, including 

liver diseases. Emerging evidence suggests that the autopagic 

functions protect hepatocytes from damages. Hence, we 

hypothesize that TSG-6 promotes the autophagical clearance 

systems in hepatocytes and protects liver from chronic injury. 

To prove this hypothesis, mice were fed with Methionine choline-deficient 

ethionine (MCDE) for 4 weeks, followed by being 

injected intraperitonially with TSG-6 (50ng) (TSG-6 group) or 

saline (MCDE+vehicle group) with MCDE supplemented diets 

for additional 2 weeks. The histomorphological liver injury and 

increased level of liver enzymes were shown in MCDE-treated 

mice with or without saline, whereas those observations were 

markedly ameliorated in TSG-6-treated mice with MCDE diet 

(AST; control: 31.32±1.74, MCDE: 198.42±7.09, MCDE+vehicle: 

184.35±26.01, MCDE+TSG-6: 140.91±31.33/ ALT; 

control: 33.95±3.09, MCDE: 151.34±40, MCDE+vehicle: 

156.14±32.34, MCDE+TSG-6:111.91±10.39). RNA analysis 

showed the up-regulation of autophagy-relate genes, atg3 and 

atg7 (2.38±0.37 fold increase for atg3, 1.8±0.17 fold increase 

for atg7; p


698 

Decellularized Spleen Matrix for Regeneration of Functional 

Hepatic-like Tissue by Reseeding Bone Marrow 

Mesenchymal Stem Cells 

Junxi Xiang 1,2 , Xinglong Zheng 1,2 , Lifei Yang 2 , Rui Gao 2 , Yi Lv 1,2 ; 

1 Department of Hepatobiliary Surgery, First Affiliated Hospital 

of Xi’an Jiaotong University, Xi’an, China; 2 Research Institute of 

Advanced Surgical Technology and Engineering, Xi’an Jiaotong 

University, Xi’an, China 

Purpose This present study intends to regenerate hepatic-like 

tissue by reseeding and inducing differentiation of bone marrow 

mesenchymal stem cells (BMSCs) in decellularized spleen 

matrix (DSM), in order to expand the donor organ pool for liver 

transplantation. Methods The spleen, which provide access to 

a wider range of alternative sources, was chosen for preparing 

decellularized scaffold. We developed a physical-chemical 

method, namely, repetitive freezing/thawing cycles, deionized 

water, trypsin and Triton X-100 perfusion, to produce rats’ 

DSM scaffold. Then, 2×10 7 BMSCs were repopulated into 

DSM for further dynamic culture and hepatic differentiation by 

using a dened two-step inducing protocol. Two-dimensional 

tissue culture asks were used as control. Results After decellularization, 

the spleen matrix presented a biomimetic three-dimensional 

porous architecture with intact vascular networks. 

The native extracellular matrix proteins including collagen I, 

collagen IV, bronectin and laminin were preserved, indicating 

a similar three-dimensional microenvironment mimicking the 

decellularized liver scaffold. When compared with two-dimensional 

culture, the produced DSM bio-scaffold promoted the cell 

engraftment and differentiation of BMSCs into hepatocyte-like 

cells using a dened protocol during a 21-day differentiation 

period, evidenced by morphological change, expression of 

hepatic-associated genes and proteins, glycogen storage, indocyanine 

green uptake, albumin secretion and urea production. 

Conclusions The physical-chemical strategy was suitable for 

producing decellularized spleen matrix, and the DSM scaffold 

might have considerable potential in cell-based therapy and 

fabricating functional hepatic-like tissue particularly because 

the DSM can support hepatic differentiation of BMSCs and 

effectively expand the donor pool. 

Disclosures: 

The following authors have nothing to disclose: Junxi Xiang, Xinglong Zheng, Lifei 

Yang, Rui Gao, Yi Lv 

699 

Wnt/beta-catenin signaling activates dUTP pyrophosphatase, 

a regulator of cellular dUTP levels, to prevent 

uracil misincorporation into DNA in human liver cancer 

stem cells 

Yoshiro Asahina, Taro Yamashita, Hajime Takatori, Naoki Oishi, 

Kouki Nio, Takehiro Hayashi, Yoshimoto Nomura, Tomoyuki 

Hayashi, Mariko Yoshida, Tomomi Hashiba, Tsuyoshi Suda, 

Masao Honda, Shuichi Kaneko; Disease Control and Homeostasis 

Internal Medicine, Kanazawa University Hospital, Kanazawa, 

Japan 

BackgroundRecent evidence suggests that hepatocellular carcinoma 

(HCC) is characterized by a hierarchical organization of 

a subset of cells termed cancer stem cells (CSCs), which may 

play a central role in resistance against cytotoxic reagents. 

However, it is still unclear, mechanistically, how CSCs prevent 

DNA damage induced by the reagents. In this study, we evaluated 

the role of dUTP pyrophosphatase (dUTPase), known to 

catalyze the hydrolysis of dUTP to dUMP, in the maintenance of 

the dUMP pool and prevention of uracil misincorporation into 

DNA in HCC CSCs. MethodsHuh7 and Huh1 cells were cultured 

routinely in DMEM supplemented with 10% fetal bovine 

serum. EpCAM-positive CSCs were purified by flow cytometry. 

The expression levels of EpCAM and dUTPase were evaluated 

by immunohistochemistry in 107 primary HCC samples that 

were surgically resected. The promoter activity of DUT (encoding 

dUTPase) was evaluated by a luciferase assay using Huh7 

cells transfected with pGL3-DUT-full and pRL-SV40 plasmids. 

The GSK3-beta inhibitor BIO, control MeBIO, and small interfering 

RNAs (siRNAs) targeting scramble or CTNNB1 sequences 

were used to regulate Wnt signaling in HCC cells. Results- 

Sorted EpCAM-positive CSCs showed more nuclear accumulation 

of dUTPase than EpCAM-negative Huh1 and Huh7 cells. 

Immunohistochemical analysis indicated a strong positive correlation 

between EpCAM and dUTPase expression in primary 

HCCs. High-dUTPase HCCs showed lower overall survival than 

that of low-dUTPase HCCs, and this difference was statistically 

significant (P = 0.0012). BIO treatment increased, whereas 

si-CTNNB1 treatment decreased the expression of EpCAM and 

dUTPase compared with that of the control Huh7 cells. We 

found a TCF4 binding element in the DUT promoter region, 

and BIO treatment enhanced, whereas si-CTNNB1 treatment 

attenuated DUT luciferase promoter activity compared with that 

of the control cells. In contrast, although 5-fluorouracil treatment 

enriched EpCAM-positive CSCs with nuclear accumulation of 

dUTPase, it had no effect on DUT promoter activity, suggesting 

no direct effect of 5-fluorouracil on DUT promoter activity. ConclusionTaken 

together, our data suggest that Wnt/beta catenin 

signaling activates dUTPase and may prevent DNA damage in 

liver CSCs. dUTPase may be a good target to eradicate liver 

CSCs resistant to cytotoxic reagents. 

Disclosures: 

Mariko Yoshida - Grant/Research Support: Bayer 






The following authors have nothing to disclose: Yoshiro Asahina, Taro Yamashita, 

Hajime Takatori, Naoki Oishi, Kouki Nio, Takehiro Hayashi, Yoshimoto Nomura, 

Tomoyuki Hayashi, Tomomi Hashiba, Tsuyoshi Suda, Masao Honda 

700 

Predictors of Response to Grazoprevir/Elbasvir Among 

HCV Genotype 1 (GT1)–Infected Patients: Integrated 

Analysis of Phase 2-3 Trials 

Stefan Zeuzem 1 , Jürgen K. Rockstroh 2 , Paul Y. Kwo 3 , David Roth 4 , 

Eric Lawitz 5 , Mark S. Sulkowski 6 , Xavier Forns 7 , Janice Wahl 8 , 

Michael Robertson 8 , Bach-Yen T. Nguyen 8 , Eliav Barr 8 , Anita Y. 

Howe 8 , Michael D. Miller 8 , Peggy Hwang 8 , Erluo Chen 8 , Kenneth 

J. Koury 8 ; 1 Department of Internal Medicine I, J.W. Goethe University 

Hospital, Frankfurt, Germany; 2 University of Bonn, Bonn, 

Germany; 3 Indiana University School of Medicine, Indianapolis, 

IN; 4 University of Miami Miller School of Medicine, Miami, FL; 

5 Texas Liver Institute, University of Texas Health Science Center, 

San Antonio, TX; 6 Johns Hopkins University School of Medicine, 

Baltimore, MD; 7 Hospital Clinic, Barcelona, Spain; 8 Merck & Co., 

Inc., Kenilworth, NJ 

Purpose: In phase 2-3 trials, 95% of GT1-infected patients 

(±cirrhosis, HIV coinfection, prior treatment, end-stage renal 

disease) who received grazoprevir 100 mg + elbasvir 50 mg 

(GZR/EBR) ± ribavirin (RBV) achieved a sustained virologic 

response 12 weeks after end of therapy (SVR12); 3% experienced 

virologic failure. This analysis assessed potential predictors 

of response to therapy. Methods: Analyses were performed 

on 2 pooled datasets of 1408 GT1-infected patients enrolled


in phase 2/3 trials of GZR/EBR ± RBV: (1) treatment-naive 

patients (TN; N=801), and (2) patients who previously failed 

peginterferon + RBV ± first-generation protease inhibitor (TE; 

N=607). Demographic factors and presence of resistance-associated 

variants (RAVs) that were fit (ie, >25% of the overall 

baseline viral load) were considered. Univariate logistic regression 

models were fitted one variable at a time in assessing 

the potential association with SVR12. Multivariable logistic 

regression (MVLR) models with forward selection were then 

applied to identify significant (ie, P < 0.1) independent predictors 

of SVR12. Results: In the MVLR, among TN patients, age, 

sex, cirrhosis, HIV coinfection, baseline NS3 RAVs, and use 

of RBV did not impact SVR12 rates. Among TN GT1a-infected 

patients, baseline HCV RNA level and baseline NS5A RAVs 

conferring >5-fold shift in the potency of EBR in vitro (termed 

NS5A >5× RAVs) were identified as significant predictors 

of SVR12. Baseline HCV RNA had a significant impact only 

when NS5A >5× RAVs were present, representing 5.3% of 

the GT1a population. No significant predictors were identified 

among GT1b-infected patients. Among TE patients, no significant 

predictors were identified for GT1b-infected patients or 

in GT1a-infected patients with prior relapse. Among GT1a-infected 

patients with prior on-treatment failure, female patients 

and noncirrhotics had higher SVR12 rates, and the addition of 

RBV and/or longer treatment durations had a positive impact 

on SVR12. The most prevalent (>1%) NS5A >5× RAVs in both 

TN and TE patients were L31M and Y93H. Baseline NS5A >5× 

RAVs, which were detected in 8% of TE GT1a-infected patients 

had a significant negative impact on SVR12; this impact was 

confined to the 12-wk treatment duration, as no patient treated 

for 16 or 18 weeks with RBV experienced virologic failure. 

Conclusion: GZR/EBR ± RBV is highly effective among GT1-infected 

patients. Among GT1a-infected patients, presence of fit 

NS5A >5× RAVs at baseline impacts efficacy. These RAVs are 

uncommon (


John M. Vierling - Advisory Committees or Review Panels: Abbvie, Bristol-Meyers-Squibb, 

Gilead, Hyperion, Intercept, Janssen, Novartis, Merck, Sundise, 

HepQuant, Salix, Immuron, Exalenz, Chronic Liver Disease Foundation; Board 

Membership: Clinical Research Centers of America, LLC; Grant/Research Support: 

Abbvie, Bristol-Meyers-Squibb, Eisai, Gilead, Hyperion, Intercept, Janssen, 

Novartis, Merck, Sundise, Ocera, Mochida, Immuron, Exalenz, Conatus; Speaking 

and Teaching: GALA, Chronic Liver Disease Foundation, ViralEd, Chronic 

Liver Disease Foundation, Clinical Care Options 




Joan R. Butterton - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: 

Merck Sharp & Dohme Corp. 



Barbara A. Haber - Employment: Merck 

702 

Short-duration therapy with daclatasvir/asunaprevir/ 

beclabuvir fixed-dose combination plus sofosbuvir in 

patients with chronic hepatitis C genotype 1 (FOURward 

Study) 

Mark S. Sulkowski 1 , Steven L. Flamm 2 , Zeid Kayali 3 , Eric Lawitz 4 , 

Paul Y. Kwo 5 , Fiona McPhee 6 , Anne Torbeyns 7 , Eric A. Hughes 8 , 

Eugene S. Swenson 6 , Philip Yin 6 , Misti Linaberry 8 ; 1 Johns Hopkins 

University, Baltimore, MD; 2 Northwestern University, Chicago, IL; 

3 Inland Empire Liver Foundation, Rialto, CA; 4 The Texas Liver Institute, 

University of Texas Health Center, San Antonio, TX; 5 Indiana 

University, Indianapolis, IN; 6 Bristol-Myers Squibb, Wallingford, 

CT; 7 Bristol-Myers Squibb, Braine l’Alleud, Belgium; 8 Bristol-Myers 

Squibb, Princeton, NJ 

Background: DCV-TRIO is a fixed-dose, twice-daily combination 

of daclatasvir 30mg (pangenotypic NS5A inhibitor), 

asunaprevir 200mg (NS3 inhibitor), and beclabuvir 75mg 

(non-nucleoside NS5B inhibitor). DCV-TRIO has achieved high 

rates of sustained virologic response at posttreatment Week 12 

(SVR12) after 12 weeks of treatment in cirrhotic and non-cirrhotic 

populations with HCV genotype (GT)1 infection. The 

FOURward study investigated the efficacy and safety of DCV- 

TRIO in combination with sofosbuvir (SOF; nucleotide NS5B 

inhibitor) for shortened treatment durations of 4 or 6 weeks. 

Methods: In this open-label, phase 2 study, non-cirrhotic treatment-naive 

patients aged ≥18 years with HCV GT1a or 1b 

infection (GT1b capped at 40%) were randomly assigned (1:1) 

to receive DCV-TRIO BID and SOF 400mg QD for 4 or 6 weeks. 

The primary endpoint was SVR12. Patients with virologic failure 

were offered 12 weeks of retreatment with DCV-TRIO+RBV 

(with optional PegIFNα) or SOF+RBV+PegIFNα-2a, dependent 

upon resistance testing by population-based sequencing at failure. 

Results: Of the 28 treated patients, 61% were female, 

89% white, 79% HCV GT1a, with a median age of 58.5 yrs. 

Median baseline viral load was high (approx. 9×10 6 IU/mL), 

61% were non-CC IL28B GT. All patients completed treatment 

and entered follow-up. HCV RNA was 2M IU/mL (7/21, 33%) vs


+ RBV (PR) ± first-generation PI. TE pts were further classified 

as those who had relapsed after completing prior treatment 

and those who experienced virologic failure (VF) on prior 

treatment (eg, null or partial responders, or virologic breakthrough). 

Results: With a 12-week regimen of GZR/EBR (no 

RBV), SVR was achieved in 94% of TN pts (5% VF), 96% of 

prior relapsers (0% VF) and 89% of prior on-treatment failures 

(10% VF). With a 16- or 18-week (+ RBV) regimen, 95% of 

prior on-treatment failures achieved SVR12 (0% VF). In each 

group, efficacy was similar in noncirrhotic and cirrhotic pts 

(Table 1). Baseline NS5A RAVs were uncommon (10%); however, 

efficacy was lower in pts with baseline NS5A RAVs conferring 

>5-fold shift in the potency of EBR in vitro, particularly 

in TN pts with high viral load and in prior on-treatment failures 

treated for 12 weeks. Conclusion: A 12-week regimen of GZR/ 

EBR (no RBV) is highly effective among GT1a-infected TN pts 

and prior relapsers. A 16-week regimen of GZR/EBR (+ RBV) is 

highly effective among TE GT1a-infected pts who experienced 

virologic failure on prior treatment. Efficacy is similar in noncirrhotic 

and cirrhotic pts. 

Table 1 

a Lost to follow-up, withdrew consent, discontinued due to adverse 

event 

b Excludes nonvirologic failures 

Disclosures: 




Abbvie, BMS 











David Roth - Advisory Committees or Review Panels: Merck, Sharp and Dome; 

Consulting: Merck, Sharp and Dome 










BMS 

Xavier Forns - Consulting: Jansen, Abbvie; Grant/Research Support: Jansen, 

Gilead 





Michael D. Miller - Employment: Merck & Co., Inc.; Stock Shareholder: Merck 

& Co., Inc. 



704 

Prevention of allograft HCV reinfection with peri-transplant 

MBL-HCV1 monoclonal antibody in combination 

with oral direct-acting antiviral 

Parvez S. Mantry 1 , Heidi L. Smith 2 , Raymond T. Chung 3 , William 

C. Chapman 4 , Michael P. Curry 5 , Thomas D. Schiano 6 , Yang 

Wang 2 , Deborah C. Molrine 2 ; 1 The Liver Institute, Methodist Dallas 

Medical Center, Dallas, TX; 2 MassBiologics of the University 

of Massachusetts Medical School, Boston, MA; 3 Department of 

Gastroenterology, Massachusetts General Hospital, Boston, MA; 

4 Department of Surgery, Washington University, St. Louis, MA; 

5 Division of Gastroenterology, Beth Israel Deaconess Medical Center, 

Boston, MA; 6 Recanati-Miller Transplantation Institute, Mount 

Sinai Medical Center, New York, NY 

Background Patients with hepatitis C viremia at the time of 

liver transplantation experience rapid infection of the donor 

allograft, increased risk of graft failure, and accelerated rates 

of fibrosis. To evaluate a peri-transplant treatment strategy to 

prevent allograft hepatitis C virus (HCV) infection, MBL-HCV1, 

a neutralizing human monoclonal antibody (mAb) targeting a 

conserved linear epitope of the HCV envelope, was combined 

with licensed direct-acting antivirals (DAA) in an open-label 

exploratory efficacy trial. Methods All subjects had HCV RNA 

titers > 10 4 IU/mL at the time of transplantation. MBL-HCV1 

was dosed intravenously at 50 mg/kg, beginning just prior to 

the anhepatic phase of the transplantation procedure. Subjects 

received three mAb infusions on the day of transplant and 

daily infusions on days 1-7 post-transplantation. Oral DAA therapy 

was initiated between days 3 and 7 with documentation 

of adequate graft function and administered in combination 

with weekly mAb infusions through day 28. During the second 

post-transplant month, MBL-HCV1 was administered bi-weekly 

while continuing oral DAA therapy; study treatment could be 

extended until day 84 (12 weeks) post-transplant in subjects 

whose HCV RNA remained undetectable. In Part 1 of the study, 

eight subjects received MBL-HCV1 in combination with telaprevir 

for up to 12 weeks. In Part 2 of the study, two subjects 

have received MBL-HCV1 in combination with sofosbuvir for 

12 weeks. Results In Part 1 of the study, prolonged aviremia 

was observed in 5 of 8 subjects and one subject (12.5%) had 

a sustained virologic response (SVR24). There were 11 serious 

adverse events assessed as unrelated to study treatment. In Part 

2 of the study, both subjects (100%) achieved SVR12 and the 

first subject has completed the 24 week visit with an SVR. One 

subject had a history of prior virologic breakthrough on interferon 

therapy, while the other previously experienced virologic 

relapse following a 12 week pre-transplant course of sofosbuvir/simeprevir. 

With MBL-HCV1 plus sofosbuvir treatment, 

both subjects were aviremic by day 21 post-transplantation. 

Two serious adverse events were reported > 2 months after 

completion of study treatment and assessed as unrelated to 

study treatment. Conclusions Peri-transplant therapy with MBL- 

HCV1 in combination with an oral DAA is capable of preventing 

post-transplant HCV recurrence and the combination 

of MBL-HCV1 with sofosbuvir appears promising. Given the 

observed viral kinetics, this treatment approach has the potential 

to achieve viral eradication in the immediate post-transplant 

period, thereby mitigating the risks of severe allograft HCV 

infection.


Disclosures: 

Parvez S. Mantry - Consulting: Salix, Gilead, Janssen, Abbvie, BMS; Grant/ 

Research Support: Salix, Merck, Gilead, Boehringer-Ingelheim, Mass Biologics, 

Vital Therapies, Santaris, mass biologics, Bristol-Myers Squibb, Abbive, Bayer-Onyx, 

Shinogi, Tacere, Intercept; Speaking and Teaching: Gilead, Janssen, 

Salix 

Heidi L. Smith - Employment: MassBiologics of UMass Medical School 


Merck, BMS 

Michael P. Curry - Advisory Committees or Review Panels: Bristol Meyers Squib, 

Abbvie; Grant/Research Support: Gilead Sciences, Mass Biologics, Merck, 

Salix, Conatus; Stock Shareholder: Achilion 

Deborah C. Molrine - Employment: MassBiologics of Univ of Massachusetts 

Medical School 

The following authors have nothing to disclose: William C. Chapman, Thomas 

D. Schiano, Yang Wang 

705 

Analysis of HCV Genotype 1 Variants Detected During 

Monotherapy and Combination Therapy with Next Generation 

HCV Direct-Acting Antiviral Agents ABT-493 and 

ABT-530 

Teresa Ng, Tami Pilot-Matias, Rakesh Tripathi, Gretja Schnell, 

Thomas Reisch, Jill Beyer, Tanya Dekhtyar, Armen Asatryan, Federico 

J. Mensa, Andrew L. Campbell, Jens Kort, Christine Collins; 

AbbVie, North Chicago, IL 

Background: ABT-530 (NS5A inhibitor) and ABT-493 (NS3/4A 

protease inhibitor identified by AbbVie and Enanta) are next 

generation HCV direct-acting antiviral agents (DAAs). Each 

has demonstrated potent anti-HCV activity in genotype (GT) 

1-infected patients in 3-day monotherapy (Study M13-595) as 

well as in combination therapy for 12 weeks (Phase 2 study 

M14-867). In this report we present the characterization of HCV 

GT1 variants detected in samples from these 2 studies. Methods: 

Population sequencing was performed on the NS3/4A 

and NS5A genes from all available baseline (BL) samples from 

both studies, last available samples with HCV RNA ≥ 1000 IU/ 

mL during monotherapy in study M13-595, and first available 

sample after virologic failure with HCV RNA ≥1000 IU/mL in 

the combination study M14-867. Sequences were examined 

for the presence of resistance-associated variants (RAVs) at 

positions where variants have been shown to confer resistance 

to HCV protease or NS5A inhibitors. NS3 or NS5A RAVs were 

introduced into appropriate subgenomic HCV GT1 replicons 

and their susceptibility to ABT-493 or ABT-530 was evaluated 

in a transient transfection assay. Results: In ABT-493 monotherapy 

study (n=49), none of the BL samples had NS3 RAVs. A 

single NS3 RAV (GT1a A156T) emerged in 1 subject during 

therapy. In ABT-530 monotherapy study (n=40), NS5A RAVs 

(M28V, Q30R, L31M, H/P58 variants, and Y93 variants) 

were present in 8 subjects at BL. Most of these were present as 

NS5A single RAVs that do not confer resistance to ABT-530. 

During monotherapy, additional RAVs emerged in 3 of these 

8 subjects, producing double RAVs that conferred a higher 

level of resistance (27- to >300-fold) to ABT-530 than either 

RAV alone. In the M14-867 combination study of ABT-493 + 

ABT-530 (n=79), 2 subjects had NS3 RAVs (R155K) and 13 

subjects had NS5A RAVs (M28V, Q30 variants, L31M, H/P58 

variants, and Y93H) at BL. To date, 78 subjects have achieved 

SVR 4 

(99% SVR 4 

) and 1 subject relapsed at post treatment 

week 4. In the relapse subject, no BL RAVs were detected, 

but a double NS5A RAV (Q30K+H58D) emerged at failure. 

Conclusions: Variants resistant to ABT-493 were not detected 

in subjects at BL, and a single NS3 RAV (A156T) emerged 

in 1 subject during ABT-493 monotherapy. During ABT-530 

monotherapy, additional NS5A variants emerged in 3 of the 8 

subjects who already had variants present at BL. These double 

RAVs conferred medium to high resistance to ABT-530. In the 

combination study, a treatment-emergent double NS5A RAV 

was detected at failure in the subject who relapsed. All other 

subjects from this study have achieved SVR 4 

, regardless of the 

presence of BL NS3 and/or NS5A variants. 

Disclosures: 


AbbVie 

Tami Pilot-Matias - Employment: AbbVie; Stock Shareholder: AbbVie 

Rakesh Tripathi - Employment: AbbVie Inc.; Stock Shareholder: AbbVie Inc. 

Gretja Schnell - Employment: AbbVie Inc.; Stock Shareholder: AbbVie Inc. 

Thomas Reisch - Employment: Abbvie; Stock Shareholder: Abbvie 

Jill Beyer - Employment: Abbvie; Stock Shareholder: Abbvie 

Tanya Dekhtyar - Employment: Abbvie; Stock Shareholder: Abbvie 



Andrew L. Campbell - Employment: AbbVie; Stock Shareholder: AbbVie 


Christine Collins - Employment: AbbVie, Inc. 

706 

Improvement in liver disease parameters following 

treatment with daclatasvir + sofosbuvir and ribavirin 

in patients with chronic HCV infection and advanced 

cirrhosis 

Robert J. Fontana 1 , Fred Poordad 2 , Eugene R. Schiff 3 , John M. 

Vierling 4 , Charles S. Landis 5 , Rafia Bhore 6 , Philip Yin 7 , Stephanie 

Noviello 6 , Eugene S. Swenson 7 ; 1 University of Michigan Medical 

Center, Ann Arbor, MI; 2 University of Texas Health Science Center, 

Texas Liver Institute, San Antonio, TX; 3 University of Miami 

Milller School of Medicine, Schiff Center for Liver Diseases, Miami, 

FL; 4 Baylor College of Medicine, Houston, TX; 5 University of Washington 

School of Medicine, Seattle, WA; 6 Bristol-Myers Squibb, 

Princeton, NJ; 7 Bristol-Myers Squibb, Wallingford, CT 

Background: The pangenotypic combination of daclatasvir 

(DCV) and sofosbuvir (SOF), with or without ribavirin (RBV) has 

achieved SVR12 rates of 82-98% in multiple high-need patient 

populations with chronic HCV infection. In the phase 3 ALLY-1 

study, DCV+SOF+RBV achieved SVR12 in 83% of patients 

with advanced cirrhosis and in 94% of those with post-liver 

transplant recurrence. We evaluated changes over time in liver 

disease parameters among patients from the advanced cirrhosis 

cohort of ALLY-1. Methods: This open-label study enrolled 

treatment-naive or -experienced adults with HCV infection of 

any genotype (GT). Patients in the advanced cirrhosis cohort 

(N=60) received 12 weeks of treatment with DCV 60 mg 

+ SOF 400 mg once-daily and RBV (initially 600 mg/day, 

adjusted for hemoglobin and creatinine clearance). The primary 

endpoint was HCV RNA


clusions: DCV+SOF+RBV treatment in patients with advanced 

cirrhosis achieved high SVR12 rates and improved clinical and 

biochemical indicators of liver disease. 

Disclosures: 








Eugene R. Schiff - Advisory Committees or Review Panels: Bristol Myers Squibb, 

Gilead, Merck, Janssen, Salix Pharmaceutical, Pfizer, Arrowhead; Consulting: 

Acorda; Grant/Research Support: Bristol Myers Squibb, Abbott / AbbVee, Gilead, 

Merck, Conatus, Medmira, Roche, Janssen, Orasure Technologies, Discovery 

Life Sciences, Siemens, Beckman Coulter, Siemens 









Charles S. Landis - Grant/Research Support: Gilead, Abbvie, BMS 

Philip Yin - Stock Shareholder: Bristol-Myers Squibb 

Stephanie Noviello - Consulting: Merck/Schering-Plough; Employment: Bristol-Myers 

Squibb, Merck/Schering-Plough; Stock Shareholder: Merck/Schering-Plough, 

J&J 

Eugene S. Swenson - Employment: Bristol-Myers Squibb 

The following authors have nothing to disclose: Rafia Bhore 

up to follow-up week 24 (FU24) will be reported at the presentation. 

Methods: The Phase 2 portion is a randomized, double-blind 

trial (unblinding occurred when all patients achieved 

FU4) to evaluate the tolerability, efficacy and pharmacokinetics 

(PK) of GZR/EBR. Non-cirrhotic treatment-naive or -experienced 

GT1 CHC patients were randomized to receive either 50 or 

100 mg of GZR co-administered with 50 mg EBR; all study 

medications were given QD for 12 weeks. Treatment response 

was assessed using COBAS TaqMan v2.0 (lower limit of quantitation 

< 1.2 Log IU/mL). Intensive PK sampling over a 24-hour 

dosing period for non-compartmental analyses was performed 

in a subset of patients at treatment week 4 (TW4). Results: 

Sixty-three patients were enrolled (42% male; mean age 61; 

100% GT1b); one subject never took study medication due 

to an abnormal ECG. Mean baseline HCV-RNA was 6.2 Log 

IU/mL. The most common adverse events (AEs) were headache, 

nausea and fatigue. There was no dose relationship for 

AEs. No patient discontinued due to an AE or study medication 

intolerance. All patients who received study medication 

(n=62) suppressed HCV-RNA to TND (Target Not Detected) 

during treatment. No patient experienced virologic breakthrough/rebound 

during treatment and all patients achieved 

SVR4; however, 1 patient in the GZR 100 mg arm relapsed 

at FU12. PK parameters (e.g., C 2hr 

, C 24hr 

and AUC 0-24 

) were 

calculated and compared with non-Japanese data from another 

GZR/EBR study that administered 100 mg GZR + 50 mg EBR. 

Conclusions: GZR/EBR therapy was well-tolerated in Japanese 

patients with GT1 CHC infection, across GZR doses. In 

addition, SVR12 was achieved in 61/62 patients (98.4%). 

Exposures of GZR and EBR were slightly higher in Japanese 

non-cirrhotic patients compared to non-Japanese non-cirrhotic 

patients. GZR 100 mg + EBR 50 mg QD was selected as the 

dose to move forward into the Phase 3 portion of the study 

based on comparable efficacy and tolerability. 

Patients with undetectable HCV-RNA 

707 

Efficacy, Safety And Pharmacokinetics Of Grazoprevir 

(MK-5172) And Elbasvir (MK-8742) In Hepatitis C Genotype 

1 Infected Non-Cirrhotic Japanese Patients (Phase 

2 Portion In Phase 2/3 Combined Study) 

Norifumi Kawada 1 , Fumitaka Suzuki 2 , Yoshiyasu Karino 3 , Kazuaki 

Chayama 4 , Yoshito Itoh 5 , Takeshi Okanoue 6 , Makoto Nakamuta 7 , 

Naoyoshi Yatsuzuka 8 , Go Fujimoto 8 , Lisa Chiacchierini Lupinacci 9 , 

Michael Robertson 9 , Luzelena Caro 9 , Daniel A. Tatosian 9 , Anita Y. 

Howe 9 , Hiromitsu Kumada 2 ; 1 Department of Hepatology, Osaka 

City University Medical School, Osaka, Japan; 2 Department of 

Hepatology, Toranomon Hospital, Tokyo, Japan; 3 Department of 

Gastroenterology, Sapporo Kosei General Hospital, Hokkaido, 

Japan; 4 Department of Gastroenterology and Metabolism, Hiroshima 

University, Hiroshima, Japan; 5 Department of Molecular 

Gastroenterology and Hepatology, Kyoto Prefectural University 

of Medicine, Kyoto, Japan; 6 Department of Gastroenterology and 

Hepatology, Saiseikai Suita Hospital, Osaka, Japan; 7 Department 

of Gastroenterology, Kyushu Medical Center, National Hospital 

Organization, Fukuoka, Japan; 8 MSD K.K., Tokyo, Japan; 9 Merck 

& Co., Inc., Kenilworth, NJ 

Background: Grazoprevir (MK-5172, GZR; NS3/4A protease 

inhibitor) and elbasvir (MK-8742, EBR; NS5A inhibitor) are 

being developed as components of an all-oral direct-acting 

antiviral regimen for the treatment of chronic hepatitis C (CHC). 

The aim of the Phase 2 portion of this study was to compare 

regimens containing 50 or 100 mg GZR + 50 mg EBR with the 

objective of assessing the tolerability and efficacy in genotype 

1 (GT1)-infected non-cirrhotic CHC Japanese patients. Results 

Disclosures: 



Fumitaka Suzuki - Speaking and Teaching: BMS 

Yoshiyasu Karino - Speaking and Teaching: BMS KK 


Astellas, Torii, Tsumura, Aska, Bayer, Zeria, Daiichi Sankyo, Dainippon Sumitomo, 

Eisai, Eli Lily, Janssen, Kowa, Mitsubishi Tanabe, MSD, Nippon Kayaku, 

Nippon Shinyaku, Otsuka, Roche, Takeda, Toray; Speaking and Teaching: 

Ajinomoto, AbbVie, Abott, Astellas, AstraZeneca, Aska, Bayer, BMS, Chugai, 

Daiichi Sankyo, Dainippon Sumitomo, Eisai, J & J, Jimro, Miyarisan, MSD, Nihon 

Kayaku, Olympus 






Naoyoshi Yatsuzuka - Employment: MSD K.K. 

Go Fujimoto - Employment: MSD K.K.; Stock Shareholder: Merck 

Lisa Chiacchierini Lupinacci - Employment: Merck 


Luzelena Caro - Employment: Merck & Co., Inc. 

Daniel A. Tatosian - Employment: Merck & Co; Stock Shareholder: Merck & Co 


Hiromitsu Kumada - Patent Held/Filed: SRL; Speaking and Teaching: Bristol-Myers 

Squibb,Pharma International, MSD, Janssen Pharma., Glaxosmithkline 

The following authors have nothing to disclose: Takeshi Okanoue, Makoto 

Nakamuta


708 

Efficacy and Safety of Co-Formulated Ombitasvir/Paritaprevir/Ritonavir 

with Ribavirin in Adults with Chronic 

HCV Genotype 4 Infection in Egypt (AGATE-II) 

Gamal E. Esmat 2 , Wahid H. Doss 3 , Roula B. Qaqish 1 , Imam 

Waked 4 , Gamal E. Shiha 5 , Ayman Yosry 3 , Mohamad Hassany 3 , 

Jennifer King 1 , Coleen Hall 1 , Rebecca Redman 1 , Niloufar Mobashery 

1 ; 1 AbbVie, Inc, North Chicago, IL; 2 Cairo University, Cairo, 

Egypt; 3 National Hepatology and Tropical Medicine Research 

Institute, Cairo, Egypt; 4 National Liver Institute, Menoufiya, Egypt; 

5 Egyptian Liver Research Institute and Hospital, Dakahliah, Egypt 

Purpose: Chronic hepatitis C virus (HCV) infection is the main 

cause of liver cirrhosis and liver cancer in Egypt and one of the 

five leading causes of death. The prevalence of HCV infection 

in Egypt is the highest in the world (10-14%) with over 90% 

infected with HCV genotype (GT) 4. In the Phase 2b PEARL-I 

study, the efficacy and safety of the direct acting antiviral 

agents (DAA) ombitasvir (OBV), a NS5A inhibitor, and paritaprevir, 

a NS3/4A protease inhibitor identified by AbbVie and 

Enanta, co-dosed with ritonavir (PTV/r) with or without ribavirin 

(RBV) were assessed in 135 patients with HCV GT4 infection 

without cirrhosis. SVR12 was 100% in both treatment naïve 

(TN) and prior interferon (IFN) and RBV treatment experienced 

(TE) patients receiving OBV plus PTV/r with RBV for 12 weeks. 

AGATE II is the first phase 3 trial to evaluate OBV/PTV/r with 

RBV in Egypt for GT4 infected subjects with and without compensated 

cirrhosis. Methods: This ongoing Phase 3, multicenter, 

open label trial enrolled 160 subjects across 5 sites in Egypt. 

Non-cirrhotic patients (n=100) received co-formulated OBV/ 

PTV/r once-daily (25 mg/150 mg/100 mg) with weight based 

RBV for 12 weeks (Arm A). Cirrhotic subjects (n=60) were randomized 

1:1 to the same regimen for either 12 or 24 weeks 

(Arms B and C; n=30/arm). The primary efficacy endpoint is 

SVR12. Safety is being evaluated by adverse event (AE) monitoring, 

laboratory testing, and other standard assessments. 

Subjects will be followed for 48 weeks post treatment. Results: 

A total of 160 noncirrhotic (Arm A, n= 101) and cirrhotic 

(Arm B, n=30, C n=29) subjects were enrolled. Approximately 

half were TE (61% prior nulls, 24% prior relapsers and 15% 

partial responders). Overall, 76% are male. The average age 

is 54 years and mean BMI 29.5 kg/m 2 , with 18% reporting a 

history of diabetes and 67% with HOMA-IR scores >3. Overall 

DAA-related treatment emergent adverse events (AEs) occurring 

in >10% of subjects were fatigue (12%) and headache (15%). 

There was 1 subject with a serious AE (SAE) of deep venous 

thrombosis deemed reasonabiy possibly related to study drug. 

There were no AEs leading to discontinuation of study drug, 

1 subject withdrew consent (Arm A) and two subjects met the 

on-treatment criteria for virologic failure (Arm B, n=1 and C, 

n=1). Conclusion: The study regimen was generally well tolerated. 

SVR12 for non-cirrhotic and cirrhotic subjects in the 12 

week Arms A and B, respectively, will be presented as well 

as the current number of on-treatment virologic failures and 

post-treatment relapses for cirrhotic subjects in arm C. 

Disclosures: 

Gamal E. Esmat - Advisory Committees or Review Panels: MSD &BMS companies, 

MSD &BMS companies, Abbvie; Grant/Research Support: Gilead Sc; 

Speaking and Teaching: Roche & GSK companies, Roche & GSK companies 

Roula B. Qaqish - Employment: AbbVie 



Mohamad Hassany - Grant/Research Support: Gilead Sc 

Jennifer King - Employment: AbbVie 

Coleen Hall - Employment: AbbVie; Stock Shareholder: AbbVie 

Rebecca Redman - Employment: AbbVie; Stock Shareholder: AbbVie 

Niloufar Mobashery - Employment: Abbvie; Stock Shareholder: abbvie 

The following authors have nothing to disclose: Wahid H. Doss, Gamal E. Shiha, 

Ayman Yosry 

709 

Baseline HCV NS5A resistance-associated variants do 

not impact SVR12 rates in non-cirrhotic and post-liver 

transplant patients with genotype 1 infection treated 

with daclatasvir and sofosbuvir with or without ribavirin 

for 12 weeks: An integrated analysis 

Fiona McPhee 1 , Dennis Hernandez 1 , Nannan Zhou 1 , Joseph 

Ueland 1 , Vincent Vellucci 1 , Sandra Hartman-Neumann 1 , Xiaoyan 

Yang 1 , Zhou Han 1 , Fei Yu 1 , Rong Yang 2 , Stephanie Noviello 2 ; 

1 Bristol-Myers Squibb, Wallingford, NJ; 2 Bristol-Myers Squibb, 

Princeton, NJ 

Background: High sustained virologic response 12 weeks posttreatment 

(SVR12) has been achieved in patients infected with 

HCV genotype (GT)1 treated with daclatasvir (DCV) and sofosbuvir 

(SOF)±ribavirin (RBV) for 12 weeks. We evaluated the 

impact of baseline NS5A variants on SVR12 rates by pooling 

individual patient data from phase 2 and 3 studies. Methods: 

Data were derived from GT1, non-cirrhotic treatment-naive 

or -experienced patients with HCV infection and post-liver 

transplant recurrence (ALLY-1 [N=41]), HIV/HCV coinfection 

(ALLY-2 [N=105]), and chronic HCV infection (AI444-040 

[N=82]). Plasma samples from 228 (182 GT1a, 46 GT1b) 

patients receiving DCV 60mg (except for HIV/HCV coinfected 

patients on boosted protease inhibitors (30mg), or efavirenz or 

neviripine (90mg)) once daily (QD) and SOF 400mg QD±RBV 

were collected at baseline. HCV RNA was isolated and the 

NS5A region amplified and sequenced. The impact of DCV-resistant 

variants (at NS5A amino acid positions 28, 30, 31, 

or 93 for GT1a and at 31 or 93 for GT1b) on SVR12 rates 

were assessed in all patients with baseline sequence (n=220); 

8 patients (6 GT1a, 2 GT1b) were excluded due to poor compliance 

(N=1), incarceration (N=1), loss to follow-up (N=1) or 

no available baseline NS5A sequence (N=5). Results: Of 220 

GT1 patients with baseline NS5A sequence, 99.1% (218/220 

[39/41 posttransplant and 179/179 non-cirrhotic patients]) 

achieved SVR12. Baseline NS5A variants at amino acid positions 

associated with DCV resistance were detected in 11.8% 

(26/220 [GT1a: 10.8% (19/176); GT1b: 15.9% (7/44)]) of 

patients. SVR12 was achieved in 100.0% (26/26) of patients 

with these NS5A variants compared with 99.0% (192/194) 

without. Observed baseline NS5A variants included M28T/V, 

Q30H/L/R, L31M, and Y93C/H/S in GT1a and L31M and 

Y93H in GT1b. One posttransplant patient had NS5A variants 

M28T+Q30R at baseline; a loss in DCV activity of 44000- 

fold was observed in-vitro with M28T+Q30R vs. a GT1a reference 

sequence without these substitutions. Two posttransplant 

patients (1 GT1a, 1 GT1b) without baseline NS5A variants 

relapsed; NS5A-Q30R+L31V emerged in the GT1a patient 

and a deletion at NS5A-P32 was observed in the GT1b 

patient. These substitutions conferred >333,333-fold DCV 

resistance in-vitro. The 5 patients without available baseline 

NS5A sequence achieved SVR12. Conclusions: In this analysis 

of patients with HCV GT1 infection (non-cirrhotic patients 

with monoinfection or HIV/HCV coinfection and posttransplant 

HCV recurrence), 11.8% had baseline DCV-resistant variants. 

SVR12 was achieved in 100% of GT1 patients with these baseline 

DCV-resistant variants even when high-level DCV-resistant 

variants were detected. 

Disclosures: 

Fiona McPhee - Employment: Bristol-Myers Squibb 

Dennis Hernandez - Employment: Bristol-Myers Squibb 

Nannan Zhou - Employment: Bristol Myers Squibb Company


Joseph Ueland - Employment: Bristol-Myers Squibb 

Fei Yu - Employment: Bristol-Myers Squibb 

Rong Yang - Employment: BMS; Stock Shareholder: BMS 



J&J 

The following authors have nothing to disclose: Vincent Vellucci, Sandra Hartman-Neumann, 

Xiaoyan Yang, Zhou Han 

710 

Drug-drug interactions between next generation direct 

acting antivirals ABT-493 and ABT-530 with cyclosporine 

or tacrolimus in healthy subjects 

Matthew P. Kosloski, Sandeep Dutta, Weihan Zhao, Jingtao Wu, 

David Pugatch, Armen Asatryan, Jens Kort, Wei Liu; AbbVie, 

North Chicago, IL 

Purpose: A next generation direct acting antiviral (DAA) combination 

of ABT-493 (NS3/4A protease inhibitor discovered 

by AbbVie and Enanta) + ABT-530 (NS5A inhibitor) is being 

developed for the treatment of chronic hepatitis C (HCV) genotype 

1-6 infection. ABT-493 + ABT-530 combination demonstrated 

high sustained virologic response rate in Phase 2 

studies. Two Phase 1, open-label studies were conducted to 

assess the pharmacokinetics, safety and tolerability when ABT- 

493 + ABT-530 is coadministered with immunosuppressants 

cyclosporine or tacrolimus. Methods: Healthy adults subjects 

received single doses of cyclosporine 100 mg (n=12) or tacrolimus 

1 mg (n=12) alone or in combination with ABT-493 

300 mg QD + ABT-530 120 mg QD. Intensive blood sampling 

for determination of cyclosporine, tacrolimus, ABT-493 

and ABT-530 concentrations was performed and pharmacokinetic 

parameters (maximum observed concentration [C max 

], 

area under the concentration-time curve [AUC t 

or AUC inf 

] 

and trough concentration [C 24 

]) were estimated. Safety and 

tolerability were assessed throughout the study. Results: Cyclosporine 

C max 

, AUC t 

, and AUC inf 

in blood were minimally 

affected (≤14% change) when coadministered with steady-state 

ABT-493 + ABT-530. Steady-state C max 

, AUC 24 

, and C 24 

in 

plasma were slightly increased for ABT-493 (30%, 37%, and 

34%, respectively) and for ABT-530 (11%, 22%, and 26%, 

respectively) when coadministered with cyclosporine. Tacrolimus 

C max 

, AUC t 

, and AUC inf 

in blood were slightly increased 

(50%, 53%, and 45%, respectively) when coadministered with 

steady-state ABT-493 + ABT-530. Steady-state C max 

, AUC 24 

, 

and C 24 

in plasma were minimally affected for ABT-493 (≤11% 

change) and for ABT-530 (≤2% change) when coadministered 

with tacrolimus. No serious adverse events were observed 

in either study. There were no patterns to the adverse events 

reported, and no new safety issues were identified. Conclusions: 

No dose adjustment is required for ABT-493, ABT-530, 

and cyclosporine when coadministered. No dose adjustment is 

required for ABT-493 and ABT-530 when coadministered with 

tacrolimus. It is recommended that subjects receiving tacrolimus 

should continue to use their current dose when initiating treatment 

with DAAs, and reduce the dose of tacrolimus if necessary 

based on therapeutic monitoring. 

Disclosures: 

Sandeep Dutta - Employment: AbbVie; Stock Shareholder: AbbVie 

Weihan Zhao - Employment: AbbVie, Inc; Stock Shareholder: AbbVie, Inc, Exxon 

Mobile, American Airlines 

Jingtao Wu - Employment: Abbvie 



Wei Liu - Employment: AbbVie 

The following authors have nothing to disclose: Matthew P. Kosloski, David 

Pugatch 

711 

Comparative Efficacy and Tolerability of Daclatasvir 

+ Sofosbuvir versus Sofosbuvir + Ribavirin in Patients 

with Chronic Hepatitis C Coinfected with HIV: A Matching-Adjusted 

Indirect Comparison 

Elyse Swallow 1 , Jinlin Song 1 , Yong Yuan 2 , Anupama Kalsekar 2 , 

Caroline Kelley 1 , Miranda Peeples 1 , Fan Mu 1 , Stephanie Noviello 

2 , James Signorovitch 1 ; 1 Health Economics and Outcomes 

Research, Analysis Group, Inc., Boston, MA; 2 Bristol-Myers 


Background and aims: To compare the efficacy and tolerability 

of daclatasvir and sofosbuvir (DCV+SOF) versus sofosbuvir 

and ribavirin (SOF+R) in patients coinfected with human immunodeficiency 

virus (HIV) and hepatitis C virus (HCV). Methods: 

A systematic literature review was conducted to identify Phase 

3 clinical trials suitable for comparison with the trial of DCV+- 

SOF in HIV/HCV-coinfected patients (ALLY-2). Two eligible trials 

of SOF+R (PHOTON-1 and PHOTON-2) were identified. 

Individual patient data from ALLY-2 were available; published 

summary data were extracted and pooled for the PHOTON 

trials. When multiple durations of SOF+R were assessed for 

the same genotype, only the arm with the FDA/EMA approved 

duration was included in the present analysis. Patients enrolled 

in ALLY-2 were subject to the inclusion and exclusion criteria 

reported in the PHOTON trials. To adjust for cross-trial differences, 

ALLY-2 patients were weighted to match all available 

summary baseline characteristics reported in both the ALLY-2 

and PHOTON trials; specifically, age, BMI, gender, race, ethnicity, 

treatment naïve, viral genotypes, HCV RNA level, IL28B 

genotype, cirrhosis status, CD4 T-cell count and combination 

anti-retroviral therapy (cART) regimens were included in the 

analysis. Sustained virologic response at week 12 post-treatment 

(SVR12), discontinuation due to adverse events (AEs), 

and rates of specific AEs reported for both SOF+R trials were 

compared between the treatments. Results: 91 of 153 patients 

from ALLY-2 who were treated with 12 weeks of DCV+SOF met 

the inclusion criteria of the PHOTON trials and were included 

in the analysis. 42 of the 497 patients from the PHOTON trials 

who were treatment-naïve, genotype 3, and treated with 12 

rather than 24 weeks of SOF+R were excluded. The SVR12 

rate was significantly higher among patients treated with DCV 

+ SOF than those treated with SOF + R both before (96.7% vs. 

84.6%; p=0.002) and after (100.0% vs. 84.6%; p


712 

Safety and Tolerability of Grazoprevir/Elbasvir in 

Patients With Chronic Hepatitis C (HCV) Infection: Integrated 

Analysis of Phase 2-3 Trials 

Geoffrey M. Dusheiko 1 , Michael P. Manns 2 , John M. Vierling 3 , K. 

Rajender Reddy 4 , Mark S. Sulkowski 5 , Paul Y. Kwo 6 , Eric Lawitz 7 , 

Deborah D. Brown 8 , Stephanie Klopfer 8 , Michael Robertson 8 , Janice 

Wahl 8 , Eliav Barr 8 , Edgar Charles 8 ; 1 Hepatology, Royal Free 

and University College, London, United Kingdom; 2 Hannover Medical 

School, Hannover, Germany; 3 Baylor College of Medicine, 

Houston, TX; 4 University of Pennsylvania, Philadelphia, PA; 5 Johns 

Hopkins University School of Medicine, Baltimore, MD; 6 ndiana 

University School of Medicine, Indianapolis, IN; 7 Texas Liver Institute, 

University of Texas Health Science Center, San Antonio, TX; 

8 Merck & Co., Inc., Kenilworth, NJ 

Purpose: In phase 2-3 studies, treatment with grazoprevir 

(GZR) 100 mg/elbasvir (EBR) 50 mg ± ribavirin (RBV) resulted 

in high rates of sustained virologic response (SVR) in HCV-infected 

patients, including those with compensated cirrhosis. 

The purpose of this analysis was to define the overall safety 

profile of GZR/EBR given for 8, 12, 16, or 18 weeks in these 

studies. Methods: Clinical adverse events and laboratory 

abnormalities reported on therapy, or within 14 days of endof-treatment 

were compared in 1795 patients (of whom 1033 

received GZR/EBR without RBV, 657 received GZR/EBR with 

RBV, and 105 received placebo). Results: A diverse population 

was enrolled: 61% were male, 13% Black/African-American, 

mean age 52.6 years (11% ≥65 years); 27% had compensated 

cirrhosis, and 18% were HIV/HCV coinfected. The overall 

safety profiles of GZR/EBR (without RBV) and placebo were 

comparable (Table 1). The addition of RBV was associated with 

more AEs. Among patients who received GZR/EBR, there were 

3 deaths (ventricular arrhythmia, strangulated hernia, motor 

vehicle accident); all 3 were unrelated to study medication. No 

patients who received placebo died. The frequency of adverse 

events was not associated with sex, age, presence of cirrhosis, 

or HIV/HCV coinfection. Late serum ALT elevations (defined 

as >5× ULN, in patients who had normal ALT values between 

treatment week [TW] 2-4) were noted in 0.8% of patients, 

generally at/after TW8. These were typically asymptomatic, 

resolving with continued therapy, scheduled end of therapy, 

or (in 3/1690, 0.18%) a protocol-mandated stop of therapy, 

and they were not associated with hyperbilirubinemia. Conclusion: 

GZR/EBR ± RBV was generally well tolerated in a large, 

diverse patient population with low SAE rate. Fewer discontinuations 

due to AEs and overall improved tolerability with lower 

reductions in hemoglobin and lower elevations in total bilirubin 

were demonstrated in the RBV-free cohort. ALT elevations 

occurring late in the course of therapy were infrequent and not 

clinically significant. 

Table 1 

Disclosures: 

Geoffrey M. Dusheiko - Advisory Committees or Review Panels: AbbVie, BMS, 

BMS, Merck; Board Membership: Gilead Sciences, Gilead Sciences 





Novartis 














BMS 












Deborah D. Brown - Employment: Merck & Co., Inc.; Stock Shareholder: Merck 

& Co., Inc 




Edgar Charles - Employment: Merck & Co. 

The following authors have nothing to disclose: Stephanie Klopfer


713 

Resistance Analysis of Treatment-Naïve and DAA-Experienced 

Genotype 1 Patients with and without Cirrhosis 

Who Received Short-Duration Treatment with Sofosbuvir/GS-5816+ 

GS-9857 

Edward J. Gane 2 , Evguenia S. Svarovskaia 1 , Robert H. Hyland 1 , 

Luisa M. Stamm 1 , Anu Osinusi 1 , Diana M. Brainard 1 , Krishna 

Chodavarapu 1 , Michael D. Miller 1 , Hongmei Mo 1 , Christian 

Schwabe 3 ; 1 Gilead Sciences Inc, Foster City, CA; 2 New Zealand 

Liver Transplant Unit, Auckland City Hospital, Auckland, New Zealand; 

3 Auckland Clinical Studies Ltd, Auckland, New Zealand 

Introduction: Pretreatment resistance associated variants (RAVs) 

have been associated with relapse following short treatment 

durations with regimens containing three or more directly 

acting antivirals (DAAs). GS-9857, a potent pan-genotypic 

HCV NS3 protease inhibitor (PI) administered with SOF/ 

GS-5816, a fixed dose combination of pan-genotypic NS5B 

and NS5A inhibitors, for 6 weeks was highly effective in treatment-naïve 

genotype (GT) 1 HCV patients without cirrhosis. 

Each compound in this three-drug combination demonstrated 

a high barrier to resistance, in vitro. The aim of this study 

was to characterize the effect of pretreatment RAVs on the 

treatment outcome and the viral resistance in patients who 

relapsed following treatment with SOF/GS-5816+GS-9857 

for 4 or 6 weeks. Methods: Treatment-naïve GT1 HCV-infected 

patients with or without cirrhosis were treated for 4 or 

6 weeks with SOF/GS-5816+GS-9857 (n=30). Treatment-experienced 

patients with or without cirrhosis who failed a 

prior multi-DAA regimen were treated for 6 weeks with SOF/ 

GS-5816+GS-9857 (n=45). NS3, NS5A and NS5B were 

amplified and deep sequenced (1% detection assay cut-off). 

All patients were sequenced pretreatment and at the time 

of virologic failure (all relapse). Results: Pretreatment deep 

sequencing analysis of NS3, NS5A and NS5B was successful 

for all 75 patients enrolled. Pretreatment NS3 and/or NS5A 

RAVs were detected in 51% (23/45) of treatment naïve and 

46% (14/30) in DAA-experienced patients. NS5B RAVs were 

detected in 1/75 (1%) of patients. Among the different treatment 

groups, SVR12 rates in patients with and without pretreatment 

RAVs were similar (Table). In the 4 week treatment arm, 

76% (11/15) of patients relapsed; no RAVs emerged in any 

of these 11 patients. In combined 6 week treatment arms, 22% 

(13/60) of patients relapsed; of these 13 patients, one treatment 

naïve patient with cirrhosis had low levels of a NS3 RAV 

emerge (V55A, 1.9%). No other NS3, NS5A, or NS5B RAVs 

emerged. Conclusions: In contrast to DAA regimens with SOF 

and other NS5A and NS3 inhibitors, pretreatment RAVs did 

not affect response to short durations of treatment with SOF/ 

GS-5816+GS-9857.Virologic failure was not associated with 

emergence of resistance. These clinical results suggest a high 

barrier to resistance with co-administration with SOF, GS-5816 

and GS-9857 as was seen in vitro with the individual agents. 

Disclosures: 







Sciences, Inc 




Sciences 




The following authors have nothing to disclose: Krishna Chodavarapu, Christian 

Schwabe 

714 

Efficacy and Safety of Ombitasvir/Paritaprevir/Ritonavir 

Co-Administered with Ribavirin in Adults with 

Genotype 4 Chronic Hepatitis C Infection and Cirrhosis 

(AGATE-I) 

Tarik Asselah 2 , Tarek I. Hassanein 3 , Roula B. Qaqish 1 , Jordan J. 

Feld 4 , Christophe Hezode 5 , Stefan Zeuzem 6 , Peter Ferenci 7 , Tami 

Pilot-Matias 1 , Yao Yu 1 , Rebecca Redman 1 , Niloufar Mobashery 1 ; 

1 AbbVie, Inc, North Chicago, IL; 2 Centre de Recherche sur l’Inflammation, 

Inserm UMR 1149, Université Paris Diderot, AP-HP 

Hôpital Beaujon, Clichy, France; 3 Southern California Liver Centers 

and Southern California Research Center, Coronado, CA; 

4 Toronto Centre for Liver Disease, University of Toronto, Toronto, 

ON, Canada; 5 Henri Mondor University Hospital, AP-HP, Université 

Paris-Est, Creteil, France; 6 J.W. Goethe University, Frankfurt, 

Germany; 7 Medical University Vienna, Vienna, Austria 

Purpose: HCV genotype 4 (GT4) represents approximately 

20% of global HCV infection. Although GT4 infection is 

more common in the Middle East and sub-Saharan Africa, 

with globalization, GT4 is now seen increasingly in Europe 

and many other countries. In the Phase 2b PEARL-I study, the 

efficacy and safety of the two direct acting antiviral agents 

(2DAA) ombitasvir (OBV), a NS5A inhibitor and paritaprevir, 

a NS3/4A protease inhibitor identified by AbbVie and 

Enanta, co-dosed with ritonavir (PTV/r) with or without ribavirin 

(RBV) were assessed in 135 subjects with HCV GT4 infection 

without cirrhosis. SVR12 was 100% in both treatment naïve 

(TN) and prior interferon (IFN) and RBV treatment experienced 

(TE) subjects receiving 2DAA+RBV for 12 weeks. This study 

extends those observations by evaluating OBV/PTV/r with 

RBV in HCV GT4-infected subjects with compensated cirrhosis. 

Methods: This ongoing Phase 3, randomized, open-label, multinational 

study (NCT 02265237) enrolled HCV GT4-infected 

TN subjects or IFN/RBV or pegIFN/RBV TE subjects with compensated 

cirrhosis. Subjects were randomized 1:1 to receive 

co-formulated OBV/PTV/r once daily (25 mg/150 mg/100 

mg) co-administered with weight based RBV for 12 (Arm A) or 

16 weeks (Arm B) with an approximately equal number of TN 

and TE subjects in each arm. A 24 week treatment arm (C) and 

an exploratory assessment in subjects who have experienced 

virologic failure with either sofosbuvir/pegIFN/RBV or sofosbuvir/RBV 

will follow. The primary objectives are to assess safety 

and SVR12 rates of these 2 DAA regimens as compared to a 

historical SVR12 rate for HCV GT4-infected subjects treated 

with pegIFN/RBV. Results: At the time of the abstract, 55 and 

56 cirrhotic subjects were randomized into Arms A and B, 

respectively. Of the 111 subjects, 48% were TN and 52% 

were TE with IFN/RBV or pegIFN/RBV (30% prior nulls, 12% 

prior relapsers and 10% partial responders). At baseline, 91% 

of subjects had a Child-Pugh score of 5, 6% of 6 and 3% of 

7. Overall, 72% are male, 78% White and 17% Black or 

African American. The mean age is 57 years and mean BMI


28 kg/m 2 , with 29% reporting a history of diabetes. Overall 

DAA-related treatment emergent adverse events (AEs) occurring 

in ≥10% of subjects were fatigue and headache (~15% each). 

5 subjects reported a total of 12 treatment-emergent serious 

AEs; 1 deemed related to study drug (manic crisis). No AE led 

to discontinuation of study drug; 1 subject withdrew consent 

and 1 subject met the on-treatment criteria for virologic failure. 

Conclusion: OBV/PTV/r with RBV for 12 and 16 weeks was 

generally well tolerated. SVR4 and SVR12 for subjects in Arms 

A and B will be presented. 

Disclosures: 










Roula B. Qaqish - Employment: AbbVie 





Gilead 







Yao Yu - Employment: Abbvie 

Rebecca Redman - Employment: AbbVie; Stock Shareholder: AbbVie 

Niloufar Mobashery - Employment: Abbvie; Stock Shareholder: abbvie 

12 weeks after end of therapy (SVR12). Results: A total of 886 

TN pts (164 black, 722 nonblack) and 650 TE pts (88 black, 

562 nonblack) with GT1, 4, or 6 infection were included in the 

analysis. Demographics and efficacy are displayed in Table 1. 

A total of 144/154 (94%) black TN pts who received 12-week 

regimens without RBV achieved SVR12. Among black TE pts 

who received 12-week/no RBV, 12-week/RBV, 16–18-week/ 

no RBV, or 16–18-week/RBV regimens, 31/33 (94%), 27/27 

(100%), 11/11 (100%), and 17/17 (100%), respectively, 

achieved SVR12. Conclusion: GZR/EBR was highly effective 

among black HCV-infected TN and TE pts with comparable 

responses observed in the nonblack population. 

Table 1 

715 

Efficacy of Grazoprevir (GZR) and Elbasvir (EBR) in 

Black HCV-Infected Patients: Results of a Pooled Analysis 

of Phase 2/3 Studies 

Philippe J. Zamor 1 , Jonathan McCone 2 , John M. Vierling 9 , Reem 

H. Ghalib 3 , Velimir A. Luketic 4 , Brian Pearlman 5 , Natarajan Ravendhran 

6 , Luis A. Balart 7 , Michael Robertson 8 , Peggy Hwang 8 , 

Bach-Yen T. Nguyen 8 , Janice Wahl 8 , Eliav Barr 8 , Rohit Talwani 8 ; 

1 Hepatology, Carolinas Medical Center, Charlotte, NC; 2 Mount 

Vernon Endoscopy and Liver Center, Alexandria, VA; 3 Texas 

Clinical Research Institute, Dallas, TX; 4 Virginia Commonwealth 

University School of Medicine, Richmond, VA; 5 Atlanta Medical 

Center, Atlanta, GA; 6 Digestive Disease Associates, Baltimore, 

MD; 7 Tulane University School of Medicine, New Orleans, LA; 

8 Merck & Co., Inc., Kenilworth, NJ; 9 Baylor College of Medicine, 

Houston, TX 

Background: Interferon-based HCV treatments are generally 

less effective in black compared with nonblack patients (pts). 

In phase 2/3 trials, the fixed-dose combination of grazoprevir 

100 mg (NS3/4A protease inhibitor) with elbasvir 50 mg 

(NS5A inhibitor) (GZR/EBR), with/without (±) ribavirin (RBV) 

had a favorable efficacy/safety profile in a diverse population 

of GT1/4/6-infected pts. We evaluated the efficacy of GZR/ 

EBR ± RBV among self-identified black pts enrolled in this program. 

Methods: Black and nonblack HCV-infected pts were 

drawn from 2 pooled datasets of phase 2/3 studies of GZR/ 

EBR: (1) treatment-naive (TN) pts and (2) treatment-experienced 

(TE) pts (who previously failed peginterferon/RBV ± first-generation 

protease inhibitor). Pts received GZR/EBR ± RBV for 12 

weeks (TN, TE) or 16/18 weeks (TE). Efficacy was measured 

as the proportion of pts who achieved a sustained virologic 

response, defined as HCV RNA below limits of quantification, 

Disclosures: 



Janssen 

Jonathan McCone - Speaking and Teaching: Schering-Plough, Roche, Schering-Plough, 

Roche, Schering-Plough, Roche, Schering-Plough, Roche 









Reem H. Ghalib - Grant/Research Support: Bristol Myers Squibb Pharmaceuticals, 

Vertex Pharmaceuticals, Janssen, Merck, Genentech, Idenix, Zymogenetics, 

Pharmasset, Anadys, Duke Clinical Research Institute, Achillion, Boehringer Ingelheim, 

Gilead Pharmaceuticals, Virochem Pharmaceuticals, Abbott, Medtronic 

Inc, Novartitis, Roche, Schering Plough, Salix, tibotec, Inhibitex, Takeda, Abbvie 

Brian Pearlman - Grant/Research Support: Merck, BI, BMS, J&J, Abbvie, Gilead; 


Natarajan Ravendhran - Grant/Research Support: Schering-Plough, Roche, Gilead, 

Bristol-Myers Squibb, Schering-Plough, Roche, Gilead, Bristol-Myers Squibb, 

Schering-Plough, Roche, Gilead, Bristol-Myers Squibb, Schering-Plough, Roche, 

Gilead, Bristol-Myers Squibb; Speaking and Teaching: onyx and otsuka, onyx 

and otsuka, onyx and otsuka, onyx and otsuka 

Luis A. Balart - Advisory Committees or Review Panels: abbvie, Genentech; 

Grant/Research Support: Merck, Genentech, Bayer, conatus, Ocera, Hyperion, 

Gilead Sciences, Bristol Myers Squibb, abbvie, Vertex, Merck, Genentech, 

Bayer, Conatus, Ocera, Hyperion, Gilead Sciences, Bristol Myers Squibb, 

Mochida, Eisai, Vertex, takeda, GI Dynamics, tobira; Speaking and Teaching: 

Merck, Merck, Merck, Merck, Abbvie, janssen 






Rohit Talwani - Employment: Merck


The following authors have nothing to disclose: Velimir A. Luketic 

716 

An integrated safety analysis of daclatasvir + sofosbuvir, 

with or without ribavirin, in patients with chronic 

HCV infection 

Charles S. Landis 1 , David R. Nelson 2 , Mark S. Sulkowski 3 , Peter 

J. Ruane 4 , Anthony M. Mills 5 , Fred Poordad 6 , David L. Wyles 7 , 

Rafia Bhore 8 , Peter Ackerman 9 , Khurram Rana 9 , Eugene S. Swenson 

9 , Stephanie Noviello 8 ; 1 University of Washington School of 

Medicine, Seattle, WA; 2 University Of Florida, Gainesville, FL; 

3 Johns Hopkins University, Baltimore, MD; 4 Ruane Medical and 

Liver Health Institute, Los Angeles, CA; 5 Anthony Mills MD, Inc, 

Los Angeles, CA; 6 Texas Liver Institute, University of Texas Health 

Sciences, San Antonio, TX; 7 University of California, San Diego, 

CA; 8 Bristol-Myers Squibb, Princeton, NJ; 9 Bristol-Myers Squibb, 

Wallingford, CT 

Background: The pangenotypic combination of daclatasvir 

(DCV) and sofosbuvir (SOF), with or without ribavirin (RBV), 

has achieved high SVRs (82%–98%) in multiple patient populations 

with HCV infection. We evaluated the safety of DCV+- 

SOF±RBV by pooling individual patient data from phase 2 and 

3 studies. Methods: Data were derived from treatment-naive 

or -experienced patients with HCV infection and advanced 

cirrhosis or post-transplant recurrence (ALLY-1), HIV/HCV coinfection 

(ALLY-2), HCV genotype (GT) 3 infection (ALLY-3), and 

chronic HCV infection (AI444-040). Overall, 679 patients 

received DCV+SOF±RBV for 8, 12, or 24 weeks. Pooled 

data were analyzed for on-treatment adverse events (AEs), 

serious AEs, AE-related discontinuations, and grade 3/4 AEs 

and lab abnormalities. Results: Patients were 67% male, 80% 

white/16% black, 62% naive; median age was 55 years and 

18% had cirrhosis, of whom half had hepatic decompensation 

(Child-Pugh class B and C). 49% of patients had HCV 

GT1a infection, 13% GT1b, 7% GT2, 30% GT3, 1% GT4, 

0.1% GT6. Overall, 98% of patients completed treatment. 

The most common AEs (any grade) were fatigue, headache, 

and nausea. Serious AEs, grade 3/4 AEs, and AE-related discontinuations 

were infrequent but more common in patients 

receiving RBV (Table); few were treatment-related. All 4 deaths 

were posttreatment and unrelated. Safety with DCV+SOF was 

comparable in patients with or without compensated cirrhosis. 

DCV+SOF+RBV patients with cirrhosis, most with hepatic 

decompensation, had slightly higher rates of serious AEs, anemia, 

and liver-related lab abnormalities than those without 

cirrhosis. There were few safety signals in this diverse population 

receiving a broad range of concomitant medications, 

including antiretroviral and immunosuppressive agents. Conclusion: 

DCV+SOF±RBV is associated with low rates of safety 

events. The presence of GT3 infection, HIV/HCV coinfection, 

advanced cirrhosis, or post-transplant HCV recurrence has minimal 

impact on safety, suggesting that DCV+SOF±RBV is a safe 

and well-tolerated treatment for a broad range of patients with 

chronic HCV infection. 

a 

ALLY-2, ALLY-3, AI444040 

b 

ALLY-1, AI444040 

c 

Includes ALLY-1 post-transplant patients with cirrhosis status 

missing 

d 

Excludes 5 patients with study drug overdoses reported as SAEs 

Disclosures: 


David R. Nelson - Advisory Committees or Review Panels: Merck; Grant/Research 

Support: Abbot, BMS, Beohringer Ingelheim, Gilead, Genentech, Merck, Bayer, 

Idenix, Vertex, Jansen 



BMS 





Anthony M. Mills - Advisory Committees or Review Panels: Gilead, ViiV, Merck; 

Grant/Research Support: Gilead, ViiV, Merck, BMS; Speaking and Teaching: 

Gilead 









Peter Ackerman - Employment: Bristol-Myers, Squibb 

Khurram Rana - Employment: Bristol-Myers Squibb 




J&J 


717 

C-EDGE TN: Impact Of 12-Week Oral Regimen Of Grazoprevir 

(GZR, MK-5172)/Elbasvir (EBR, MK-8742) On 

Patient-Reported Outcomes (PROs) In Treatment-Naïve 

Patients With Chronic Hepatitis C Virus (HCV) Genotype 

(GT) 1, 4, Or 6 Infection 

Jean Marie Arduino, Yang Wang, Deborah D. Brown, Shazia 

Khawaja, Elisa Martinez, Joan R. Butterton, Michael Robertson, 

Chizoba Nwankwo, T. Christopher Mast; Merck & Co., Inc., 

Kenilworth, NJ 

Background:Chronic HCV infection negatively impacts patients’ 

health and well-being.A Phase 3, double-blind, placebo-control, 

randomized trial of an oral fixed-dosed combination of GZR 

100 mg/EBR 50 mg once daily for 12 weeks was conducted 

among HCV GT1-,4-,or 6-infected patients.GZR/EBR was highly 

effective (SVR12:94.6%(95% CI: 91.5,96.8%)) and generally 

well-tolerated,with a similar safety profile to placebo.Our aim 

was to evaluate whether HCV treatment with GZR/EBR altered 

PROs.Methods:Subjects completed 5 PROs at baseline, treatment 

week 4 (TW4),TW12, follow-up week 4(FW4):SF-36v2®, 

EQ-VAS,FACIT-Fatigue Scale,Chronic Liver Disease Question-


naire-HCV(CLDQ-HCV) and Work Productivity and Activity 

Impairment.421 patients were randomized and received ≥1 

dose of study drug(GZR/EBR:n=316,Placebo:n=105).Mean 

change from baseline in PRO scores (95% CIs) by treatment 

group and treatment group differences in mean change scores 

(95%CIs) were estimated.Results:At TW12, in general, GZR/ 

EBR had more favorable changes than placebo [Figure].GZR/ 

EBR group had mean improvement (95% CIs exclude 0) in 

general and HCV-specific HRQOL. Placebo group had mean 

improvement in HCV-specific HRQOL, but a mean decline in 

Social Functioning (SF).Treatment group differences favored 

GZR/EBR over placebo in MCS (2.33 (95%CI:0.28,4.37)) 

and SF (7.25 (95%CI:2.47,12.04)). At FW4, for GZR/EBR 

(data not shown),mean improvements in general and HCV-specific 

HRQOL,fatigue levels,and activity impairment were 

observed.Treatment group differences favored GZR/EBR over 

placebo in Role Limitations-Physical(4.97(95%CI:0.17,9.77)),- 

General Health(4.94(95%CI:1.30,8.59)),S- 

F(6.97(95%CI:2.38,11.57)) and overall health(EQ-VAS 

5.42(95% CI:1.87,8.97)).Conclusion:Treatment with GZR/ 

EBR had a positive impact on PROs compared to placebo.In 

addition, changes in PROs were substantially more favorable 

than the large declines in PROs historically associated with 

interferon and ribavirin-containing regimens. 

Disclosures: 

Jean Marie Arduino - Employment: Merck & Co., Inc 

Yang Wang - Employment: Merck 

Deborah D. Brown - Employment: Merck & Co., Inc.; Stock Shareholder: Merck 

& Co., Inc 

Shazia Khawaja - Employment: Merck 




Chizoba Nwankwo - Employment: Merck 

T. Christopher Mast - Employment: Merck Research Laboratories 

The following authors have nothing to disclose: Elisa Martinez 

718 

Characterization of HCV Resistance from a 3-Day 

Monotherapy Study of GS-9857, a Novel Pangenotypic 

NS3/4A Protease Inhibitor 

Eric Lawitz 2 , Hadas Dvory-Sobol 1 , Jenny C. Yang 1 , Luisa M. 

Stamm 1 , James Taylor 1 , Diana M. Brainard 1 , Michael D. Miller 1 , 

Hongmei Mo 1 , Maribel Rodriguez-Torres 3 ; 1 Gilead Sciences, Foster 

City, CA; 2 The Texas Liver Institute, University of Texas Health 

Science Center, San Antonio, TX; 3 Fundacion de Investigacion, 

Rio Pedras, PR 

Background: GS-9857 is a pangenotypic hepatitis C virus 

(HCV) NS3/4A protease inhibitor (PI) with potent antiviral 

activity against HCV genotypes (GT) 1-6 and improved 

coverage of GT1 NS3 resistance associated variants (RAVs) 

associated with other HCV PIs, in preclinical replicon studies. 

In a 3-day monotherapy study in patients infected with HCV 

GT 1, 2, 3 or 4, GS-9857 was well tolerated and resulted 

in maximal mean viral load reduction >3 log 10 

IU/mL at the 

100mg dose across all genotypes. This analysis characterizes 

the HCV NS3 sequence data from this phase 1b study. Methods: 

The full-length NS3 gene was amplified and successfully 

deep sequenced using MiSeq for 58 patients at baseline and 

53 patients post-baseline using a 1% cutoff for reporting. NS3 

RAVs were defined as amino acid substitutions historically 

associated with resistance to HCV PIs, including positions 36, 

41, 43, 54, 55, 80, 122, 155, 156, 168 and 170 of the NS3 

protease gene of HCV GT1. Results: Baseline HCV NS3 RAVs 

were present in the HCV of 37% (9/24) of patients with GT1a, 

17% (1/6) of patients with GT1b and 16% (3/19) of patients 

with GT3; there were no baseline NS3 RAVs present in patients 

with GT2 (n=5) or GT4 (n=4). Mean maximal viral load reductions 

over 3 days of GS-9857 administration were similar in 

patients with and without baseline NS3 RAVs. The table summarizes 

the number of patients with treatment-emergent (TE) 

NS3 RAVs by genotype. Overall, by deep sequencing analysis, 

the majority of patients (74%, 39/53) did not have TE NS3 

RAVs, and there were no TE NS3 RAVs in patients with GT2 

or 4. A156T or A156V were the most prevalent substitutions 

in patients with GT1a or 1b infection and were not observed 

in patients with GT3 infection. One GT3 patient had low levels 

of A156P (2.3%). Conclusions: The presence of NS3 RAVs at 

baseline had no impact on treatment response to 3 days of 

monotherapy with GS-9857 in this phase 1b study. The lack of 

selection of NS3 RAVs in the majority of patients demonstrates 

an improved resistance profile of GS-9857. 

Number of Patients with Treatment Emergent (TE) NS3 RAVs Following 

3 Days Monotherapy with GS-9857 by Deep Sequencing 

Analysis 

Disclosures: 







Janssen, AbbVie, Bristol Meyers Squibb



Sciences 



James Taylor - Employment: Gilead Sciences 


Sciences 




Maribel Rodriguez-Torres - Advisory Committees or Review Panels: Bristol-Myers 

Squibb, Janssen R&D Ireland; Consulting: Glaxo Smith Kline, Janssen R&D Ireland, 

Theravance; Grant/Research Support: Merck, Bristol-Myers Squibb, Merck, 

Pfizer, Gilead, Johnson & Johnson, Beckman Coulter, Theravance 

719 

Pharmacokinetics of Coadministration of Pan-Genotypic, 

Direct Acting Antiviral Agents, ABT-493 and ABT- 

530, with or without Ribavirin for 12 weeks in HCV 

Infected Subjects without Cirrhosis 

Chih-Wei Lin, Wei Liu, Armen Asatryan, Stanley Wang, Federico 

J. Mensa, Jens Kort, Sandeep Dutta; Abbvie, North Chicago, IL 

Purpose: A direct acting antiviral agent (DAA) combination of 

ABT-493 (NS3/4A protease inhibitor discovered by AbbVie 

and Enanta) and ABT-530 (NS5A inhibitor) is being developed 

for the treatment of chronic hepatitis C (HCV) genotype 

(GT) 1-6 infection. In two Phase 2 studies (SURVEYOR-1 and 

-2), the ABT-493 and ABT-530 combination has demonstrated 

high sustained virologic response rates in GT1-, GT2- and 

GT3-infected subjects without cirrhosis following 12-weeks of 

treatment. Pharmacokinetics of ABT-493 and ABT-530 with or 

without ribavirin (RBV) were evaluated in these studies. Methods: 

Two open-label, multicenter studies, SURVEYOR-1 and 

-2, were conducted evaluate the efficacy, safety, and pharmacokinetics 

of co-administration of ABT-493 (200 or 300 

mg QD) and ABT-530 (40 or 120 mg QD) with or without 

RBV in GT1-, GT2- or GT3-infected subjects. Blood samples for 

pharmacokinetic analysis were collected throughout the study 

treatment period. ABT-493 and ABT-530 pharmacokinetics following 

a single dose (Day 1) and at steady state (Week 4) 

were assessed by non-compartmental methods. Results: A total 

of 274 subjects received ABT-493 and ABT-530 with or without 

RBV. Both ABT-493 and ABT-530 showed rapid absorption 

with Tmax ranging from 2 to 4 hours. Steady state ABT-493 

exposure (area under the curve from 0 to 4 hour) following 

300 mg was 2570 ng.h/mL, approximately 3.7-fold of the 

exposure following 200 mg administration. Coadministration 

of either 40 mg or 120 mg ABT-530 each with 200 mg ABT- 

493 resulted in ABT-530 exposure of 157 or 372 ng.h/mL, 

respectively. ABT-493 300 mg increased 120 mg ABT-530 

exposure by an additional 20% to 30%. Minimal accumulation 

in ABT-493 or ABT-530 exposure was observed at Week 4 

compared to Day 1. ABT-530 had minimal impact on ABT-493 

exposures, however, ABT-493 200 mg and 300 mg increased 

120 mg ABT-530 exposures to 3- to 4-fold of ABT-530 exposures 

when administered alone. HCV genotype and RBV coadministration 

had no impact on ABT-493 or ABT-530 exposures. 

Conclusions: ABT-493 exhibited non-linear pharmacokinetics 

with more than dose-proportional increase in exposures with 

increasing doses, while ABT-530 exposures increased in an 

approximately dose-proportional manner when coadministered 

with ABT-493. ABT-530 had minimal impact on ABT- 

493, while ABT-493 increased ABT-530 exposures, with the 

increase in ABT-530 exposure being dependent on ABT-493 

dose. ABT-493 or ABT-530 had minimal accumulation in exposures 

following multiple dosing in HCV-infected subjects. 

Disclosures: 








720 

Daclatasvir exposure does not explain lower sustained 

virologic response rates in cirrhotic patients with HCV 

genotype 3 following 12 weeks of daclatasvir plus 

sofosbuvir treatment 

Timothy Eley, Reena Wang, Shu-Pang Huang, Yash Gandhi, 

Brenda Cirincione, Frank LaCreta, Tushar Garimella; Bristol-Myers 


Background: The ALLY-3 study evaluated daclatasvir (DCV; 60 

mg daily) plus sofosbuvir (SOF; 400 mg daily) for 12 weeks in 

treatment-naive and -experienced patients with HCV genotype 

3 (GT 3) infection. Sustained virologic response at posttreatment 

week 12 (SVR12) was lower in patients with compensated 

cirrhosis than in those without (63% [20/32] vs 96% 

[105/109]), mostly due to posttreatment relapse. Pharmacokinetic 

(PK) exposure to DCV in ALLY-3 was evaluated in patients 

with and without cirrhosis to explore this observation. Methods: 

Systemic exposure to total (protein-bound + unbound) DCV on 

dosing Day 1 was evaluated in a prespecified 24-hour intensive 

PK substudy of cirrhotic GT 3 patients in ALLY-3 (N=10) 

using noncompartmental methods. DCV exposure parameters 

were compared to those of non-cirrhotic ALLY-3 patients derived 

from population PK (PopPK) modeling, and against historical 

single-dose DCV data in HCV-uninfected subjects with hepatic 

impairment. Model-predicted steady-state average DCV concentrations 

across the dosing interval (C avgss 

) for cirrhotic and 

non-cirrhotic ALLY-3 patients with and without SVR12 were 

compared visually in an exploratory exposure-response assessment. 

Results: Taking into account the historical DCV accumulation 

ratio in HCV-infected patients (1.2–1.4), median observed 

Day 1 AUC 0-24h 

in cirrhotic GT 3 patients in ALLY-3 (6210 

[range 3141–10895] ng.h/mL) was comparable to PopPK 

model estimates of median steady-state AUC 0-24h 

in both cirrhotic 

patients (7980 [3744–17856] ng.h/mL) and non-cirrhotic 

patients (9192 [3696–19776] ng.h/mL). Similarly, after 

considering the accumulation, the median observed Day 1 

AUC 0-24h 

in cirrhotic patients was comparable to historical single-dose 

median AUC inf 

for DCV 30 mg in HCV-uninfected 

subjects with Child-Pugh class A, B or C hepatic impairment 

normalized to 60 mg (9638–11398 ng.h/mL). Calculated 

median C avgss 

exposures were similar in cirrhotic and non-cirrhotic 

ALLY-3 patients in the PopPK model (non-cirrhotic: 358 

ng/mL without SVR12 [n=4], 408 ng/mL with SVR12 [n=105]; 

cirrhotic 283 ng/mL without SVR12 [n=12], 382 ng/mL with 

SVR12 [n=20]). Despite a weak trend towards lower C avgss 

in 

patients without SVR12 there was substantial overlap between 

those who did and those who did not achieve SVR12 in both 

groups, though numbers were small. Conclusions: The data 

indicate that higher relapse rates among GT 3-infected cirrhotic 

patients in ALLY-3 cannot be explained by DCV exposure 

alone. These data suggest that cirrhotic patients with HCV GT 3 

may require longer (>12 weeks) treatment duration with DCV+- 

SOF or the addition of ribavirin. 

Disclosures: 

Timothy Eley - Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers 

Squibb


Shu-Pang Huang - Employment: Bristol-Myers Squibb Company; Stock Shareholder: 

Bristol-Myers Squibb Company 

Yash Gandhi - Employment: Bristol-Myers Squibb 

Brenda Cirincione - Employment: BMS 

Frank LaCreta - Employment: Bristol-Myers Squibb; Management Position: Bristol-Myers 

Squibb; Stock Shareholder: Bristol-Myers Squibb 

Tushar Garimella - Employment: Bristol Myers-Squibb; Stock Shareholder: Abbvie, 

Bristol Myers-Squibb 

The following authors have nothing to disclose: Reena Wang 

721 

Pharmacokinetics of Narlaprevir an NS3/4A Protease 

Inhibitor with or without Ritonavir Following Single 

Dose Use in Patients with Compensated Cirrhosis and 

Healthy Volunteers 

Dmitry Koloda 1 , Natalia Tikhonova 1 , Irina Malaya 2 , Miroslav 

Ryska 3 , Mikhail Samsonov 1 , Vasily Isakov 4 ; 1 Medical Department, 

R-Pharm, Moscow, Russian Federation; 2 Ascent Clinical Research 

Solutions, Moscow, Russian Federation; 3 Quinta Analytica, 

Prague, Czech Republic; 4 Department of gastroenterology & hepatology, 

Institute of Nutrition, Moscow, Russian Federation 

Background and Aims: Narlaprevir (NVR), a potent ritonavir-boosted 

inhibitor of HCV NS3 protease, is currently being 

developed for therapy of chronic HCV genotype 1 infection 

in Russia and CIS countries. The purpose of this study was to 

evaluate the pharmacokinetics (PK) after a single oral dose of 

NVR alone and in combination with ritonavir (RTV) in healthy 

controls and in patients with compensated cirrhosis. Methods: 

This was a two-part, open-label, parallel group, single-dose 

phase I study in patients with compensated cirrhosis Child-Pugh 

class A (CPA) (n=16), and matched healthy subjects (n=16), 

all Caucasians. In Part 1 of the study 8 healthy adult subjects 

and 8 patients with CPA received single dose of NVR 200 

mg under fed condition. In Part 2 of the study 8 healthy subjects 

and 8 CPA patients received NVR 100 mg (dose was 

decreased based on the Part 1 results) in combination with RTV 

100 mg. Noncompartmental model was used for PK analysis of 

NVR in plasma and urine. Simulation of PK parameters for NVR 

at steady state was performed. Geometric least square means 

(GMEAN) and 90% confidence intervals were calculated. 

Results: In Part I GMEAN C max, 

and AUC (0-inf) 

were 1.6 and 2.7 

times higher in CPA patients vs. healthy subjects, respectively 

(Table 1). Simulated steady-state (tau=24 h) GMEAN C max, 

and 

AUC tau, 

in patients with CPA were 167% and 237% vs. healthy 

subjects respectively (Table 2). Increased NVR exposure in 

patients with CPA was well tolerated and not associated with 

increased adverse events rate. There were no significant difference 

in plasma NVR exposure in CPA patients and healthy subjects 

after single dose NVR 100 mg in combination with 100 

mg RTV and after steady-state simulation (tau=24 h): GMEAN 

C max, 

and AUC tau, 

were 107% and 105% in patients with CPA 

vs. healthy subjects, respectively (Table 2). Conclusions: NVR 

exposure after a single dose 200 mg was higher in patients 

with CPA than in healthy subjects. No significant effect on 

NVR exposure in CPA subjects compared to healthy subjects 

was found when NRV 100 mg was co-administered with RTV 

100 mg. 

Disclosures: 

Dmitry Koloda - Employment: R-Pharm 

Natalia Tikhonova - Employment: R-Pharm 

Mikhail Samsonov - Employment: RPharm 

Vasily Isakov - Advisory Committees or Review Panels: Abbvie, Bristol-Myers 

Squibb, Gilead, Janssen, Merck, Vertex, R-Pharm; Consulting: Bristol-Myers 

Squibb, Merck; Speaking and Teaching: Bristol-Myers Squibb, Janssen, Merck 

The following authors have nothing to disclose: Irina Malaya, Miroslav Ryska 

722 

Ombitasvir/Paritaprevir/r and Dasabuvir with Ribavirin 

for HCV Genotype 1 Patients with Decompensated Cirrhosis 

Parvez S. Mantry 1 , John Hanson 2 , Roger Trinh 3 , Alnoor Ramji 4 , 

Linda Fredrick 3 , Manal Abunimeh 3 , Leticia Canizaro 3 , Li Liu 3 , 

Nancy Shulman 3 , Stuart C. Gordon 5 ; 1 The Liver Institute at Methodist 

Dallas, Dallas, TX; 2 Charlotte Gastroenterology and Hepatology, 

PLLC, Charlotte, NC; 3 AbbVie, Inc., North Chicago, IL; 

4 University of British Columbia, Vancouver, BC, Canada; 5 Henry 

Ford Health System, Detroit, MI 

Objective: Patients with HCV and decompensated cirrhosis are 

at greatest risk for life-threatening complications leading to 

liver transplantation. The 3 direct-acting antiviral (3D) regimen 

of ombitasvir (OBV) co-formulated with paritaprevir/ritonavir 

(PTV/r, identified by AbbVie and Enanta) and dasabuvir (DSV) 

with ribavirin (RBV) has high efficacy in HCV genotype (GT) 

1-infected patients with compensated cirrhosis. We assessed 

the safety, pharmacokinetics, and efficacy of the 3D + RBV 

regimen in an initial cohort of HCV GT1-infected patients with 

decompensated cirrhosis. Methods: Patients with decompensated 

cirrhosis (Child-Pugh score 7 – 9) and the following laboratory 

parameters at the time of screening were enrolled: 

platelet count ≥25 x 10 9 /L, serum albumin ≥2.8 g/dL, MELD 

score ≤18. Those with GT1b or GT1a infection received standard 

dosing of OBV/PTV/r (25/150/100 mg/day) and DSV 

(250 mg twice daily) plus weight-based RBV (1000 – 1200 

mg/day) for 12 or 24 weeks, respectively. A data review 

was planned once the initial cohort reached week 12 of treatment, 

prior to enrollment of a larger cohort. Results: Baseline 

characteristics of the 11 patients enrolled are detailed in the 

Table. Suppression of HCV RNA below the level of quantitation 

occurred in 7/11 (64%) patients by treatment week 2 and in 

all patients by week 6. At treatment week 12, all 11 patients 

remained HCV RNA suppressed below the level of detection. 

Sustained virologic response at post-treatment week 12 will


be presented. No patients prematurely discontinued from the 

study, experienced a treatment-emergent adverse event (AE) 

leading to interruption of study drug (Table), or experienced 

post-baseline ALT elevations. Grade 3 bilirubin elevations, 

predominantly indirect, have occurred in 7/11 patients. The 

median starting RBV dose was 1200 mg. Hemoglobin declines 

below 10 g/dL occurred in 4 patients, 3 of whom modified 

RBV dose; none interrupted study drug. Cross-study comparison 

revealed that the range of PTV/r, OBV, DSV and DSV metabolite 

exposures observed were comparable or slightly higher 

than those observed in patients with Child-Pugh A cirrhosis. 

Conclusions: In this initial cohort to assess safety of the 3D + 

RBV regimen in HCV GT1-infected patients with decompensated 

cirrhosis, treatment was generally well tolerated. Further 

assessment of this regimen in patients with decompensated cirrhosis 

is underway with additional GT1-infected patients who 

will be treated with 3D + RBV, and GT4-infected patients who 

will be treated with OBV/PTV/r + RBV. 

Disclosures: 





Salix 

John Hanson - Advisory Committees or Review Panels: Abbvie, Gilead; Grant/ 

Research Support: Abbvie, Takeda, Salix, Gilead, Celltrion, Genentech; Speaking 

and Teaching: Janssen, Abbvie, Prometheus, Takeda 

Alnoor Ramji - Advisory Committees or Review Panels: Roche, Merck, Gilead, 

Janssen, Boehringer Ingelheim; Grant/Research Support: BMS, Roche, Merck, 

Gilead, Vertex, Novartis, Abbvie, Boehringer Ingelheim 

Linda Fredrick - Employment: AbbVie; Management Position: AbbVie; Stock 

Shareholder: AbbVie 

Manal Abunimeh - Employment: AbbVie 

Nancy Shulman - Employment: Abbvie 




The following authors have nothing to disclose: Roger Trinh, Leticia Canizaro, 

Li Liu 

723 

Absence of significant drug-drug interactions between 

next generation direct acting antivirals ABT-493 and 

ABT-530 and methadone or buprenorphine/naloxone 

in subjects on opioid maintenance therapy 

Matthew P. Kosloski, Sandeep Dutta, Weihan Zhao, Armen Asatryan, 

Jens Kort, Wei Liu; AbbVie, North Chicago, IL 

Purpose: A next generation direct acting antiviral (DAA) combination 

of ABT-493 (NS3/4A protease inhibitor discovered by 

AbbVie and Enanta) + ABT-530 (NS5A inhibitor) is being developed 

for the treatment of chronic hepatitis C (HCV) genotype 

1-6 infection. ABT-493 + ABT-530 combination demonstrated 

high sustained virologic response rate in Phase 2 studies. A 

Phase 1, open-label study was conducted to assess the pharmacokinetics, 

safety and tolerability of ABT-493 + ABT-530 and 

methadone or buprenorphine/naloxone. Methods: Otherwise 

healthy adults subjects on individualized regimens of methadone 

(n=12) or buprenorphine/naloxone (n=12) for opioid 

addiction received ABT-493 300 mg QD + ABT-530 120 mg 

QD for 7 days. Intensive blood sampling for determination 

of methadone, buprenorphine, norbuprenorphine, naloxone, 

ABT-493 and ABT-530 concentrations was performed and 

pharmacokinetic parameters (maximum observed concentration 

[C max 

], area under the concentration-time curve [AUC 24 

or 

AUC t 

] and trough concentration [C 24 

]) were estimated. Safety 

and tolerability were assessed throughout the study. Potential 

opioid withdrawal or overdose symptoms (pharmacodynamics) 

were assessed with validated instruments including the short 

opiate withdrawal scale, desire for drugs questionnaire, and 

pupillometry measurements throughout the study. Results: For 

subjects on methadone maintenance therapy, dose-normalized 

C max 

, AUC 24 

, and C 24 

for R- and S-methadone were unaffected 

by coadministration with ABT-493 and ABT-530 at steady-state 

(≤5 % change). For subjects on buprenorphine/naloxone maintenance 

therapy, dose-normalized C max 

, AUC 24 

, and C 24 

were 

slightly increased for buprenorphine (8%, 17%, and 24%, 

respectively) and norbuprenorphine (25%, 30%, and 21%, 

respectively) when coadministered with ABT-493 and ABT-530 

at steady-state; naloxone dose-normalized C max 

and AUC t 

were 

minimally affected (≤12% change). Pharmacodynamics of the 

methadone or buprenorphine/naloxone regimens were not 

significantly impacted by coadministration with ABT-493 or 

ABT-530 for either regimen. ABT-493 and ABT-530 exposures 

following coadministration with methadone or buprenorphine/ 

naloxone were similar to the observed values in previous studies. 

Subjects experienced adverse events of mild intensity, with 

the most common (reported in >5 subjects) being abdominal 

pain, constipation, and headache; all subjects completed the 

study. There were no clinically relevant abnormal laboratory 

abnormalities, ECG or vital sign findings. Conclusions: No 

dose adjustments are required for coadministration of ABT-493 

and ABT-530 with methadone or buprenorphine/naloxone. 

No pharmacodynamic interaction is expected. 

Disclosures: 


Weihan Zhao - Employment: AbbVie, Inc; Stock Shareholder: AbbVie, Inc, Exxon 

Mobile, American Airlines 




The following authors have nothing to disclose: Matthew P. Kosloski


724 

A Pilot Evaluation of Twice Daily Fixed Dose Combination 

Asunaprevir + Daclatasvir + Beclabuvir ± Weight 

Based Ribavirin in Treatment-Naïve, Non-cirrhotic 

Patients with Chronic Genotype 1a Hepatitis- C for 

Eight, Six or Four Weeks (RHACE-1) 

Gregory J. Botwin 2 , Aliya Asghar 2 , Deval Modi 2 , Dean Bosman 2 , 

Timothy R. Morgan 1,3 ; 1 Gastroenterology (11), VA Long Beach 

Healthcare System, Long Beach, CA; 2 Research Service, VA Long 

Beach Healthcare System, Long Beach, CA; 3 Division of Gastroenterology, 

University of California - Irvine, Irvine, CA 

Background: The all oral, fixed-dose combination of daclatasvir, 

asunaprevir, and beclabuvir (collectively FDC) without ribavirin 

(RBV) achieved sustained virologic response (SVR 12 

) rates of 

>92% in genotype 1, treatment naïve, non-cirrhotic patients 

treated for 12 weeks in a Phase II trial. Purpose: To assess 

the feasibility of an 8, 6 and/or 4 week treatment duration 

with FDC ± weight based RBV in treatment-naïve, non-cirrhotic, 

HCV genotype 1a subjects. Methods: We designed a single 

site, open-label, multi-arm, adaptive clinical trial and planned 

to enroll 12, non-cirrhotic, genotype (GT)-1a, treatment-naïve 

subjects per arm. To minimize the number of subjects exposed 

to a potentially ineffective treatment regimen, only one arm was 

to be enrolled at a time. Subjects enrolled in the first arm, Arm 

A, were to be treated for 8 weeks with FDC + RBV. If a low 

relapse rate was observed, then sequentially shorter treatment 

durations of 6 and 4 weeks ± RBV would be tested. The results 

presented here are for Arms A and B. Results: Arm A (n=12) 

had an average age of 60 years, BMI of 28, and baseline 

HCV RNA of 6.5 log10 IU/mL. All 12 subjects were male, 7 

were non-CC IL28B, and 3 were African-American (AA). All 12 

subjects had HCV RNA less than the limit of detection (LOD) at 

the end of treatment (Week 8). Five subjects experienced post 

treatment viral relapse and the remaining 7 (58%) achieved 

SVR 12 

. Due to the non-optimal relapse rate in Arm A, Arm B 

was adapted to 12 weeks of treatment with RBV and no other 

arms were enrolled. Arm B (n=12) had an average age of 60 

years, BMI of 27, and baseline HCV RNA of 6.6 log10 IU/ 

mL. All 12 subjects were male, 10 were non-CC IL28B, and 4 

were AA. To date all subjects in Arm B have reached the end 

of treatment (Week 12) and all have HCV RNA < LOD. Half 

of the subjects in each arm experienced a >2.5g/dL drop in 

hemoglobin on-treatment. There were no treatment-related serious 

adverse events in either group. Conclusion: Eight weeks, 

or less, of treatment with FDC plus RBV is likely to result in a 

non-optimal SVR 12 

amongst non-cirrhotic, GT-1a subjects. Less 

than 12 weeks of treatment with FDC may still be feasible in 

easier to treat populations. SVR 12 

results will be available for 

subjects enrolled in Arm B. 

Viral Response 

† One subject was excluded (n=11) 

‡ Missing data on one subject, counted as failure. 

Note: Number and percent of subjects with HCV RNA


Dennis Wolford - Employment: Merck Sharpe & Dohme Corp., a subsidiary of 

Merck and Co., Inc.; Stock Shareholder: Merck Sharpe & Dohme Corp., a subsidiary 

of Merck and Co., Inc. 

William L. Marshall - Employment: Merck 

Marian Iwamoto - Employment: Merck Sharp & Dohme Corp. 



The following authors have nothing to disclose: Katherine M. Dunnington, Nadia 

Cardillo Marricco, Michael Gartner, Dennis Swearingen, John Brejda, Angela 

O. Choi 

726 

Integrated Safety Analysis of Daclatasvir Plus Sofosbuvir, 

With or Without Ribavirin, in Patients With HCV 

Genotype 3 Infection 

David Bernstein 1 , Charles S. Landis 2 , Eric Lawitz 3 , Anne Luetkemeyer 

4 , Melissa Harris 5 , Rafia Bhore 5 , Eugene S. Swenson 6 , 

Peter Ackerman 6 , Khurram Rana 6 , Douglas Dieterich 7 ; 1 Hofstra 

North Shore-Long Island Jewish School of Medicine, Manhasset, 

NY; 2 University of Washington School of Medicine, Seattle, WA; 

3 Texas Liver Institute, University of Texas Health Science Center, 

San Antonio, TX; 4 University of California and San Francisco General 

Hospital, San Francisco, CA; 5 Bristol-Myers Squibb, Princeton, 

NJ; 6 Bristol-Myers Squibb, Wallingford, CT; 7 Icahn School of Medicine 

at Mount Sinai, New York, NY 

Background: SVR12 rates of up to 98% have been achieved 

in the three phase 3 ALLY studies with the pangenotypic combination 

of daclatasvir (DCV) plus sofosbuvir (SOF), with or 

without ribavirin (RBV). In patients with genotype (GT)3 infection 

enrolled in these studies, SVR12 rates were 88–100%. The 

safety profile of DCV+SOF±RBV in patients with GT3 infection 

was evaluated using integrated data from the ALLY studies. 

Methods: Integrated data were derived from 182 GT3-infected 

patients enrolled in ALLY-1 (advanced cirrhosis or post-liver 

transplant recurrence, N=17), ALLY-2 (HIV-HCV coinfection, 

N=13), and ALLY-3 (GT3 infection, N=152). Patients were 

treated with DCV 60mg (dose-adjusted for concomitant antiretroviral 

therapy in HIV coinfected patients) +SOF 400mg 

for 8 (ALLY-2) or 12 weeks (ALLY-2, ALLY-3) or with DCV+- 

SOF+RBV for 12 weeks (ALLY-1). Integrated safety data were 

analyzed for on-treatment adverse events (AEs), serious AEs 

(SAEs), AE-related discontinuations, and grade 3/4 AEs and 

lab abnormalities. Results: Patients were 64% male, 91% 

white, and 64% HCV treatment-naive; the median age was 

55 years and 21% had cirrhosis. In this cohort, there were 

no treatment-related SAEs or deaths. There were 7 grade 3/4 

AEs and 16 grade 3/4 lab abnormalities. Four AEs led to 

discontinuation of study drug (all treatment-related): 3 events 

were associated with RBV and led to discontinuation of only 

RBV in patients with advanced cirrhosis; 1 event (headache) 

in a liver transplant recipient led to discontinuation of all HCV 

therapy after 4 weeks, but SVR12 was still achieved. Overall, 

the most common AEs (any grade) in the ALLY studies 

were fatigue (19%), headache (17%), and nausea (12%). As 

expected, safety events were more frequent in patients with 

advanced cirrhosis that received RBV; however, none of these 

patients required discontinuation of all study therapies and 

5/6 achieved SVR12. Conclusions: Safety events with DCV+- 

SOF±RBV were uncommon in patients with HCV GT3 infection. 

HIV coinfection had no notable impact on safety parameters; 

events were more frequent in patients with advanced cirrhosis 

that received RBV, but had minimal impact on SVR12. These 

results support the use of DCV+SOF±RBV therapy in a broad 

spectrum of patients with GT3 infection. 

Disclosures: 

David Bernstein - Advisory Committees or Review Panels: Gilead; Consulting: 

Abbvie, BMS, Merck, Janssen; Grant/Research Support: Gilead, Abbvie, BMS, 

Merck, Janssen, Genentech; Speaking and Teaching: Abbvie, BMS, Merck, 

Gilead 









Anne Luetkemeyer - Grant/Research Support: Gilead, Abbvie, Pfizer, Bristol 

Myers Squibb, Merck 

Melissa Harris - Employment: Bristol-Myers Squibb 



Khurram Rana - Employment: Bristol-Myers Squibb 




727 

Projected Long-Term Impact of Grazoprevir (GZR, 

MK-5172)/Elbasvir (EBR, MK-8742) in Treatment-Naive 

and Treatment-Experienced Patients with Hepatitis C 

Virus Genotype 1 Infection and Chronic Kidney Disease 

Shannon A. Ferrante, Elamin Elbasha, Wayne Greaves, Chizoba 

Nwankwo; Merck & Co., Inc, North Wales, PA 

Background: Hepatitis C virus (HCV) infection is an important 

cause of morbidity, liver disease-related deaths from complications 

of decompensated cirrhosis (DC) and hepatocellular 

carcinoma (HCC), and cardiovascular mortality in chronic kidney 

disease (CKD) patients in the United States. GZR (NS3/4A 

protease inhibitor) and EBR (NS5A inhibitor) were recently 

studied in C-SURFER, a phase 2/3 double-blind, placebo-control 

trial in HCV genotype 1 patients with CKD4/5. Aims: To 

translate short-term findings from the C-SURFER trial into longterm 

predictions of the impact of GZR/EBR on incidence of liver-related 

morbidity and mortality compared with no treatment 

(NoTX) and pegylated interferon plus ribavirin (peg-IFN/RBV). 

Methods: A computer-based mathematical model of the natural 

history of chronic HCV genotype 1 infection and chronic 

kidney and liver disease was developed to project lifetime 

cumulative incidence of DC, HCC, liver-transplant (LT) and 

liver-related death. Efficacy of GZR/EBR was obtained from 

C-SURFER trial. In the pre-specified primary population, the 

proportion of patients achieving sustained viral response 12 

weeks after the completion of therapy was 0.99 (0.95–1.00). 

Based on the results of a meta-analysis, we assumed an efficacy 

of 0.60 (0.47–0.71) for peg-IFN/RBV. Data on baseline 

characteristics of the simulated patients were obtained from 

the National Health and Nutrition Examination Survey. Natural 

history parameters were estimated from published studies.


We estimated base case values for incidence and conducted 

sensitivity analyses. Results: GZR/EBR was projected to reduce 

lifetime cumulative incidence of DC to 3.8% from 22.0% and 

11.0% compared with NoTX and Peg-IFN/RBV, respectively 

(Table 1). The incidence of HCC was projected to be 26.7% 

in patients who received NoTx and 11.2% with peg-IFN/ 

RBV compared with 1.1% when GZR/EBR was used. As a 

result, GZR/EBR reduced liver-related mortality from 35.7% 

with NoTX and 14.3% with Peg-IFN/RBV to 0.3%. GZR/EBR 

extended life expectancy by 7.6 and 3.0 years compared with 

NoTx and peg-IFN/RBV, respectively. The results were sensitive 

to assumed patient characteristics. Conclusion: Use of GZR/ 

EBR was projected to substantially reduce the incidence of 

liver-related complications and mortality in treatment-naïve and 

treatment-experienced patients with hepatitis C virus genotype 

1 infection and chronic kidney disease. 

Table 1. Lifetime incidence of liver complications and life expectancy 

by regimen 

Disclosures: 

Shannon A. Ferrante - Employment: Merck & Co, Inc 

Elamin Elbasha - Employment: Merck & Co., Inc; Stock Shareholder: Merck & 

Co., Inc 

Wayne Greaves - Employment: Merck 

Chizoba Nwankwo - Employment: Merck 

728 

Daclatasvir exposure alone does not explain HCV 

relapse in HIV-HCV coinfected patients receiving daclatasvir 

plus sofosbuvir with ritonavir-boosted darunavir in 

the ALLY-2 study 

Tushar Garimella, Yash Gandhi, Reena Wang, Peter Ackerman, 

Zhaohui Liu, Joseph J. Raybon, Brenda Cirincione, Frank LaCreta, 

Timothy Eley; Bristol-Myers Squibb Research and Development, 

Princeton, NJ 

Background: ALLY-2 evaluated daclatasvir (DCV; 30, 60 or 

90 mg daily, adjusted for type of antiretroviral therapy) plus 

sofosbuvir (SOF; 400 mg daily) for 12 or 8 weeks in HCV 

treatment-naive or -experienced patients coinfected with HIV-1 

and HCV genotypes 1- 4. Sustained virologic response at 

posttreatment week 12 (SVR12) was lower after 8 weeks of 

DCV+SOF than after 12 weeks (76% vs 97%); mostly due to 

relapse. Three-quarters (9/12) of all relapsers were receiving 

ritonavir-boosted darunavir (DRV/r). In ALLY-2, the standard 

DCV dose of 60 mg QD was reduced a priori to 30 mg QD 

when coadministered with permissible ritonavir-boosted protease 

inhibitors, based on results from a drug-drug interaction 

(DDI) study showing ≈2-fold increase in DCV exposure with 

boosted atazanavir (ATV/r). Subsequent data from a dedicated 

DDI study in healthy subjects (HS) indicate that a DCV 

dose-adjustment is unnecessary when used with darunavir/ 

ritonavir (DRV/r) or lopinavir/ritonavir (LPV/r); the standard 60 

mg dose is now recommended. Thus, the relationship between 

DCV exposure and SVR12 was explored in patients receiving 

DRV/r in ALLY-2. Methods: Composite plasma C trough 

concentration 

values in ALLY-2 patients were derived from individual 

DCV C trough 

data at dosing Weeks 1, 2, 4, 8 and 12 (for the 

12-week treatment arms). Data between those who did or did 

not receive DRV/r and did or did not achieve SVR12 were 

visually compared. Population PK (PopPK)-modeled DCV exposures 

for ALLY-2 patients were then compared with previously 

observed HS DDI data. Results: SVR12 among those on DRV/r 

in the 12-week arms was 93% (28/30), comparable to other 

regimens in the study, versus 67% (14/21) in the 8-week arm. 

Although a trend towards lower DCV C trough 

was observed 

in patients on DRV/r who did not achieve SVR12, there was 

substantial overlap with those who did achieve SVR12 on 

DRV/r (82% [42/51] of all patients on DRV/r) or LPV/r (100% 

[12/12]). DCV trough concentrations for DRV/r and LPV/r 

were comparable in the 12- and 8-week arms, and comparable 

to patients receiving 60 mg DCV. PopPK data for ALLY-2 

patients was consistent with HS data for boosted PIs. Baseline 

HCV RNA was >2 million IU/mL in 15/21 DRV/r patients 

in the 8 week arm, including all 7 relapsers. The two DRV/r 

relapsers in the 12-week arms each had baseline HCV RNA 

>10 million IU/mL and cirrhosis. Conclusions: Relapse among 

patients receiving DRV/r with 30 mg DCV in ALLY-2 may be 

associated primarily with shorter (8-week) treatment duration 

and high baseline HCV RNA, and cannot be solely explained 

by slightly lower DCV exposures. DCV should be dosed at 60 

mg daily with DRV/r or LPV/r based on new DDI data. 

Disclosures: 

Tushar Garimella - Employment: Bristol Myers-Squibb; Stock Shareholder: Abbvie, 

Bristol Myers-Squibb 

Yash Gandhi - Employment: Bristol-Myers Squibb 


Brenda Cirincione - Employment: BMS 

Frank LaCreta - Employment: Bristol-Myers Squibb; Management Position: Bristol-Myers 

Squibb; Stock Shareholder: Bristol-Myers Squibb 

Timothy Eley - Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers 

Squibb 

The following authors have nothing to disclose: Reena Wang, Zhaohui Liu, 

Joseph J. Raybon 

729 

C-EDGE Co-Infection: Impact Of 12-Week Oral Regimen 

Of Grazoprevir (GZR, MK-5172)/Elbasvir (EBR, 

MK-8742) On Patient-Reported Outcomes (PROs) In 

Treatment-Naïve Patients With HCV/HIV Co-infection 

Jean Marie Arduino, Zhiwei Jiang, Melissa Shaughnessy, Shazia 

Khawaja, Elisa Martinez, Heather L. Platt, Chizoba Nwankwo, T. 

Christopher Mast; Merck & Co., Inc., Kenilworth, NJ 

Background: HCV/HIV co-infection negatively impacts patients’ 

health and well-being. Treatment with interferon (IFN) and ribavirin 

(RBV) further diminishes patient-reported health states. 

This international, open-label, single arm study evaluated the 

fixed-dosed combination of GZR 100 mg/EBR 50 mg once 

daily for 12 weeks among treatment- naïve patients with HCV 

GT1-,4-, or 6- co-infection with HIV. GZR/EBR was highly effective 

(SVR12: 95.0% (95% CI: 91.2%, 97.5%)) and generally 

well-tolerated. Our aim was to evaluate whether treatment 

with GZR/EBR altered PROs during treatment and follow-up. 

Method: Subjects completed the following PROs at baseline, 

treatment week 4 (TW4), TW12, and follow-up week 4 (FW4): 

SF-36v2® Health Survey, EuroQol-Visual Analogue Scale 

(EQ-VAS), FACIT-Fatigue Scale, HCV-specific Chronic Liver Disease 

Questionnaire (CLDQ-HCV) and Work Productivity and 

Activity Impairment due to Hepatitis C. Mean change from 

baseline in the PRO scores, with 95% CIs, were estimated. 

Results: Mean improvement from baseline scores (95% CIs 

exclude 0) at TW12 occurred in general (Bodily Pain, General 

Health, Vitality, Mental Component Summary (MCS), EQ-VAS) 

and HCV-specific HRQOL [Figure]. The largest mean improvements 

occurred in General Health and Vitality domains. These 

improvements were also reflected in mean improvements in


fatigue levels. In addition, patients reported considerably less 

activity impairment (


mately 10 hours (Day -1) and 2 hours (Day 1) prior to SD 100 

mg GZR/50 mg EBR FDC on Day 1. In Period 3, 40 mg PAN 

was administered QD on Days 1 to 5 followed by administration 

of SD 100 mg GZR/50 mg EBR FDC 2 hours after PAN 

administration on Day 5. There was at least a 10-day washout 

between periods. PK parameters were determined for GZR and 

EBR on Day 1 of Periods 1 and 2, and on Day 5 of Period 3. 

Results Coadministration of FAM 10 and 2 hours prior to FDC 

of GZR/EBR did not have a clinically relevant effect on SD 

GZR AUC0-∞, Cmax, and C24, with geometric mean ratios 

(GMRs) [90% confidence intervals (CIs)] of 1.10 [0.95, 1.28, 

0.89 [0.71, 1.11], and 1.12 [0.97, 1.30], respectively. FAM 

also did not have a clinically relevant effect on EBR AUC0-∞, 

Cmax, and C24, with GMRs [90% CIs] of 1.05 [0.92, 1.19], 

1.11 [0.98, 1.26], and 1.03 [0.91, 1.17], respectively. Similarly, 

coadministration of PAN with GZR/EBR FDC did not 

have a clinically relevant effect on SD GZR AUC0-∞, Cmax, 

and C24, with GMRs [90% CIs] of 1.12 [0.96, 1.30], 1.10 

[0.89, 1.37], and 1.17 [1.02, 1.34], respectively. PAN also 

did not have a clinically relevant effect on EBR AUC0-∞, Cmax, 

and C24, with GMRs [90% CIs] of 1.05 [0.93, 1.18], 1.02 

[0.92, 1.14], and 1.03 [0.92, 1.17], respectively. Conclusions 

Coadministration of GZR/EBR FDC with FAM or PAN 

had no clinically relevant effect on the PK of GZR or EBR. These 

results demonstrate that acid-reducing agents, such as H2-receptor 

antagonist and proton-pump inhibitors, may be coadministered 

with the GZR/EBR FDC in HCV-infected patients 

without restriction. 

Disclosures: 

Hwa-Ping Feng - Employment: Merck 

Pavan Vaddady - Employment: Merck & Co., Inc. 

Deborah L. Panebianco - Employment: Merck 

Luzelena Caro - Employment: Merck & Co., Inc. 



Marian Iwamoto - Employment: Merck Sharp & Dohme Corp. 

Wendy W. Yeh - Employment: Merck 

The following authors have nothing to disclose: Patrice Auger, Xiaobi Huang, 

Fang Liu, Chun Feng, Nadia Cardillo Marricco, Vanessa Levine, David Goblet, 

Michael Gartner, Daria Stypinski 

732 

Viral suppression with IFN-free therapies ameliorates 

portal hypertension in patients with hepatitis C-related 

cirrhosis 

Mattias Mandorfer, Karin Kozbial, Philipp Schwabl, Clarissa Freissmuth, 

Remy Schwarzer, Rafael Stern, Simona Bota, Thomas 

Reiberger, Albert Stättermayer, Wolfgang Sieghart, Sandra Beinhardt, 

Michael Trauner, Harald Hofer, Arnulf Ferlitsch, Peter Ferenci, 

Markus Peck-Radosavljevic; Department of Internal Medicine 

III, Medical University of Vienna, Vienna, Austria 

Background: Interferon (IFN)-free regimens are highly effective 

and generally well tolerated. Portal pressure, assessed by 

hepatic venous pressure gradient (HVPG) measurement, drives 

the development of liver-related events and mortality. Since a 

decrease in HVPG translates into an immediate clinical benefit, 

it is an excellent surrogate endpoint. Aim: To investigate the 

effect of IFN-free treatments on portal pressure in patients with 

hepatitis C-related cirrhosis. Methods: Our study comprises 25 

patients with hepatitis C-related cirrhosis and portal hypertension 

(HVPG ≥6mmHg) at baseline (BL) who underwent HVPG 

measurement after 12-24 weeks of sofosbuvir-based IFN-free 

therapy (FU). Concomitant medication known to have an effect 

on HVPG, including but not limited to beta blockers, were 

paused prior to the HVPG measurements. Results are reported 

as median[interquartile range] or mean±standard deviation. 

Results: The majority of patients had Child-Pugh stage (CPS) 

A (80%[20/25]) and compensated cirrhosis (92%[23/25]). 

Ascites was the reason for decompensation in both patients 

and none of the patients had CPS C cirrhosis. Forty-eight percent 

(12/25) of patients had varices (small: 67%[8/12]/large: 

33%[4/12]) and two patients (8%[2/25]) had a history of variceal 

bleeding. All patients had undetectable HCV-RNA at the 

time of FU HVPG measurement. Antiviral therapy significantly 

decreased HVPG (BL: 16[10.5] vs. FU: 13[11.5]mmHg; mean 

of differences: -2.4±3.15mmHg; P20% or 

to ≤12mmHg) was observed in 40%(6/15). Sixty-one percent 

(11/18) of patients achieved a clinically meaningful HVPG 

decrease defined as (A) in patients with CSPH but without varices, 

or (B) in patients with a HVPG >12mmHg and varices. 

Conclusions: While Afdhal and co-workers did not observe an 

effect of viral suppression on HVPG in their study population 

comprised of a higher proportion of decompensated CPS B/C 

patients, effective antiviral therapy led to an immediate and 

clinically meaningful decrease in HVPG in our study. Thus, 

IFN-free therapies are likely to become a cornerstone in the 

treatment of portal hypertension, especially in patients with 

compensated cirrhosis. The results will be updated to include 

additional patients and information on potential predictors of 

HVPG-response. 

Disclosures: 




Squibb 











Harald Hofer - Advisory Committees or Review Panels: Gilead, Abbvie; Speaking 

and Teaching: Janssen, BMS, Gilead, Abbvie 








The following authors have nothing to disclose: Karin Kozbial, Philipp Schwabl, 

Clarissa Freissmuth, Remy Schwarzer, Rafael Stern, Albert Stättermayer, Sandra 

Beinhardt, Arnulf Ferlitsch


733 

Propranolol for Prevention of Recurrence of Varices 

After Endoscopic Eradication 

Housam Helmy 1 , Hassan E. Zaghla 1 , Wael Abdel-Razek 1 , Mohammed 

Abbasy 1 , Hasan A. Elzohry 1 , Ashraf Y. El-Fert 2 , Enas Korayem 

3 , Gamal. A. Badra 1 , Imam Waked 1 ; 1 Hepatology, National 

Liver Institute, Menoufiya, Egypt; 2 Biochemistry, National Liver Institute, 

Shebeen El Kom, Egypt; 3 Radiology, National Liver Institute, 

Shebeen El Kom, Egypt 

Background and aim: Acute variceal bleeding is a life-threatening 

complication of portal hypertension associated with mortality. 

Endoscopic band ligation can eradicate varices without 

an effect on portal pressure. Non-selective beta-blockers are 

effective in reducing portal pressure and preventing primary 

and secondary variceal bleeding. Whether beta-blockers can 

prevent recurrence of varices after eradication by band ligation 

is not settled. The purpose of this study was to evaluate 

whether propranolol can prevent the recurrence of varices after 

endoscopic eradication. Methods: Ninety patients in whom varices 

were eradicated by band-ligation (70 to prevent variceal 

re-bleeding and 20 for primary prophylaxis) were randomized 

to receive for 24 months the maximum tolerable dose of propranolol 

(n=47) or follow-up (n=43). Patients were excluded 

if they had hepatocellular or extrahepatic malignancy, portal 

vein thrombosis, refractory ascites, hepatorenal syndrome, 

Child-Turcotte-Pugh (CTP) class C, advanced systemic disease, 

or contra-indication to propranolol. All enrolled patients were 

followed endoscopically every 3 months. The primary endpoint 

was the recurrence of varices, and varices that recurred were 

re-ligated. Results: Both groups were comparable regarding 

age (propranolol: 46.4±11.2, follow-up: 50.1±7.3, P=0.07), 

gender [propranolol: males: 29 (61.7%), follow-up: 27 

(62.8%), P=0.915] and history of variceal bleeding [propranolol: 

35 (74.5%), follow-up: 35 (81.4%), P=0.459). The median 

dose of propranolol was 40 (20-120) mg/day. Median CTP 

and MELD scores did not differ between groups at baseline 

(CTP: 5 vs. 5, P=0.272; MELD: 6 vs. 6, P=0.840) or at the 

end of follow-up (CTP: 6 vs. 5.5, P=0.280; MELD: 6.43 vs. 6, 

P=0.264). Number of patients with recurrence of varices [propranolol: 

31 (66%), follow-up: 35 (81.4%), P=0.098] did not 

differ between groups. However, time to recurrence of varices 

was significantly longer in the propranolol group (20.9±1.2 

months vs. 15.3±1.2 months, P=0.008). No severe adverse 

events, rebleeding or mortality occurred during the study and 

no patients needed drug discontinuation. Conclusion: Propranolol 

use safely and significantly delayed, but did not reduce, 

the recurrence of esophageal varices. 

Disclosures: 



The following authors have nothing to disclose: Housam Helmy, Hassan E. 

Zaghla, Wael Abdel-Razek, Mohammed Abbasy, Hasan A. Elzohry, Ashraf Y. 

El-Fert, Enas Korayem, Gamal. A. Badra 

734 

Factors Influencing The First Decompensation Of Cirrhosis 

With Portal Hypertension And Varices In Primary 

Prophylaxis With Β-Blockers 

Edilmar A. Alvarado 1 , Oana Pavel 1 , Alba Ardevol 1 , Miguel 

Martinez 1 , Natali Escajadillo 1 , Miguel Rios 1 , Maria Poca 1 , Mar 

Concepción 1 , Torras Xavier 1,2 , Carlos Guarner 1,2 , Càndid Villanueva 

1,2 ; 1 Hospital de la santa creu i sant pau, Barcelona, Spain; 

2 Centro de investigación Biomédica en red en el Area temática de 

Enfermedades Hepáticas y Digestivas, Barcelona, Spain 

Patients with clinically significant portal hypertension (CSPH) 

and varices have a higher risk of decompensation.Hepatic 

venous pressure gradient (HVPG) response induces a reduction 

in risk of variceal bleeding.Factors such as HVPG non-response, 

obesity or alcohol have been associated with higher risk of 

decompensation.However, determinants of decompensation in 

patients with CSPH and varices have not been clarified.The 

aim of this study was to investigate the main determinants of 

decompensation in compensated cirrhosis treated with β-blockers 

for primary prophylaxis. METHODS: Cirrhotic patients 

referred for primary prophylaxis, were included.Hepatic 

and systemic hemodynamic assessment was performed.Portal 

pressure was estimated by HVPG measured before and 

after i.v. administration of propranolol (0.15 mg/kg).Nadolol 

was administered to prevent bleeding.Parameters which 

may influence the development of decompensation such as 

acute response to β-blockers(βB), obesity, metabolic syndrome 

or hepatic function,were evaluated in patients without previous 

decompensation of cirrhosis. RESULTS: 285 patients were 

included. Alcohol and HCV were the most common ethiology 

(33%,44%) and 161(56%)/103(36%)/ 21(7%)were Child- 

Pugh class A/B/C.In acute β-blockers test 49% of patients had 

a decrease HVPG ≥10% from baseline,137(48%)patients had 

no previous decompensation.Among compensated patients,decompensation 

occurred in 89(59%)after a mean follow-up of 

48±38 months:73(53%)had ascites,22(16%)SBP,11(8%)hepatorenal 

syndrome,22(16%)variceal bleeding,37(27%)encephalopathy,26(19%)hepatocellular 

carcinoma and 40(29%)died. 

Acute response to BB was the parameter which better discriminate 

decompensation,that occurred in 45% of responders vs 

71% of non-reponders(P= 0.01) while death occurred in 19% 

vs 43% respectively(P< 0.01).Cox regression analysis, acute 

hemodynamic response(HR= 2.4, 95% CI= 1.3-4.1),Child- 

Pugh score(HR= 1.9, 95% CI= 13-2.6) and active alcohol 

intake(HR= 2.2, 95% CI= 1.1-5.2) independently predicted the 

development of decompensation, while obesity, metabolic syndrome 

and MELD did not.Acute hemodynamic response(HR= 

2.5, 95% CI= 1.2-5.2) and development of decompensation(HR= 

2.4, 95% CI= 1.0-6.9) independently predicted mortality. 

CONCLUSIONS: In compensated cirrhosis on primary 

prophylaxis with BB, acute hemodynamic response is a strong 

predictor of decompensation while Child-Pugh and active alcoholism 

also have independent prognostic value. Acute hemodynamic 

response and decompensation independently predict 

mortality.These results suggest that etiologic therapies and 

treatments to improve hemodynamic response can be prevent 

decompensation and improve survival. 

Disclosures: 

The following authors have nothing to disclose: Edilmar A. Alvarado, Oana 

Pavel, Alba Ardevol, Miguel Martinez, Natali Escajadillo, Miguel Rios, Maria 

Poca, Mar Concepción, Torras Xavier, Carlos Guarner, Càndid Villanueva


735 

Obesity is associated with increased mortality and need 

for transplant in patients with alcoholic liver disease 

Mohamed G. Shoreibah 2 , Donny Kakati 1 , Sudha Kodali 2 , Ashwani 

K. Singal 2 ; 1 Internal Medicine, University of Alabama at Birmingham, 

Birrmingham, AL; 2 Gastroenetrology and Hepatology, 

University of Alabam at Birmingham, Birmingham, AL 

Obesity is associated with increased mortality and need for 

liver transplant in patients with alcoholic liver disease Background 

and aims: Alcoholic liver disease (ALD) is the second 

most common cause of cirrhosis and the need for liver transplant. 

Increasing prevalence of obesity has also resulted in a 

growing population of obese alcoholics. Data on the impact of 

obesity on the natural history of ALD are scarce. Methods: We 

performed a retrospective study on 286 well characterized ALD 

patients. After excluding 28 patients without body mass index 

(BMI) data at presentation, 258 ALD patients were stratified 

into three groups based on BMI as Group A: normal weight 

(BMI40 g/d in women and >60 

g/d in men for >5 years. Results: Group A (N=97; mean age 

47) differed compared to overweight alcoholics (N=78; mean 

age 46 y) and obese alcoholics (N=83; mean age 43 y) for 

male gender (53% vs. 58% vs. 70%; P


737 

Clinical and histologic correlates of the hepatic venous 

pressure gradient (HVPG) in patients with compensated 

cirrhosis due to nonalcoholic steatohepatitis (NASH) 

Arun J. Sanyal 1 , Zachary D. Goodman 2 , Manal F. Abdelmalek 3 , 

Stephen A. Harrison 4 , Don C. Rockey 5 , Anna Mae Diehl 3 , Stephen 

H. Caldwell 6 , Mitchell L. Shiffman 7 , Robert P. Myers 8 , Raul E. 

Aguilar Schall 8 , Mani Subramanian 8 , John G. McHutchison 8 , Vlad 

Ratziu 9 , Nezam H. Afdhal 10 , Jaime Bosch 11 ; 1 Virginia Commonwealth 

University, Richmond, VA; 2 Inova Fairfax Hospital, Falls 

Church, VA; 3 Duke Clinical Research Institute, Durham, NC; 4 Fort 

Sam Houston, San Antonio Military Medical Center, San Antonio, 

TX; 5 Medical University of South Carolina, Charleston, SC; 6 University 

of Virginia, Charlottesville, VA; 7 Liver Institute of Virginia, 

Richmond, VA; 8 Gilead Sciences, Inc., Foster City, CA; 9 Hôpital 

Pitié-Salpêtrière, Paris, France; 10 Beth Israel Deaconess Medical 

Center and Harvard Medical School, Boston, MA; 11 University of 

Barcelona, Barcelona, Spain 

Background: Clinical complications in patients with cirrhosis 

are predominantly linked to the severity of portal hypertension. 

Our objective was to examine clinical and histologic correlates 

of the HVPG in patients with compensated cirrhosis due to 

NASH. Methods: The study included adults with compensated 

cirrhosis (Ishak 5 or 6) due to NASH who were enrolled in a 

phase 2b trial of simtuzumab, a monoclonal antibody against 

lysyl oxidase-like-2 (LOXL2). Liver biopsies were graded centrally 

according to the NAFLD Activity Score (NAS) and hepatic 

collagen was quantified via computer-assisted morphometry. 

HVPG was measured according to a standardized protocol 

and reviewed centrally. Serum LOXL2 (sLOXL2) was measured 

using an immunoassay (VIDAS® LOXL2; bioMérieux, Marcy 

L’Etoile, France). The associations between HVPG and age, 

sex, body mass index (BMI), diabetes, use of non-selective beta 

blockers (NSBBs), esophageal varices, laboratory variables, 

fibrosis stage, hepatic collagen, and NAS and its elements 

were determined. Stepwise-forward, logistic regression models 

evaluated independent predictors of clinically significant 

portal hypertension (CSPH, defined as an HVPG ≥10 mmHg). 

Results: 230 of 258 randomized patients (89.1%) with HVPG 

and complete histologic data were included. The median age 

was 56 years (IQR 51-61), 63% were female, 67% had stage 

6 fibrosis, 40% had a NAS ≥5, and the median hepatic collagen 

content was 12.5% (n=218 with complete data; IQR 

8.1-19.4%). The median HVPG was 12 mmHg (IQR 9-16.5) 

and 69% had CSPH. The HVPG was weakly correlated with 

hepatic collagen content (Spearman ρ=0.19; P=0.004) and 

moderately correlated with sLOXL2 (ρ=0.50; P


Demographic data were comparable between both groups. 

After a mean follow-up of 17.3±13.1 months in group A, 11 

patients (23.4%) bled and 13 patients (27.7%) died. In group 

B, 6 patients bled (12.2%) and 9 patients (18.4%) died after 

a mean follow-up of 18.9±14.2 months. The rebleeding and 

survival rates analyzed by the Kaplan-Meier method with logrank 

test were not significantly different between both groups 

(P=0.09 and 0.3, respectively). Age was the independent factor 

of rebleeding (OR: 1.1, 95% CI: 1.0 – 1.1, P=0.03). Hepatoma 

(OR: 5.6, 95% CI: 2.3 – 13.3, P


740 

Hemostatic profiles in liver disease: novel evidence of 

mixed phenotype in critically-ill patients with cirrhosis 

and liver failure and implications for clinical management 

of hemostatic disturbance. 

Vishal C. Patel 1 , Arjuna Singanayagam 1 , Jelle Adelmeijer 2 , Julia 

Wendon 1 , William Bernal 1 , Ton Lisman 2 ; 1 Liver Intensive Therapy 

Unit, Kings College Hospital, London, United Kingdom; 2 Surgical 

Research Laboratory, University Medical Center Groningen, Groningen, 

Netherlands 

Background Hemostatic disturbance is ubiquitous in critically-ill 

patients with cirrhosis and liver failure but there is lack of 

evidence to guide management. Studies in patients with compensated 

cirrhosis suggests a ‘rebalanced’ or pro-thrombotic 

state, but very limited data exists as to that present in those with 

decompensated disease or acute on chronic liver failure. We 

performed detailed evaluation of hemostatic profiles in patients 

with liver failure of varying severity, comparing to those in 

healthy controls. Patients and Methods Hemostatic profiles 

were determined in patients on admission to a single centre 

with stable (SC, n=4: median age 55 yrs (IQR 42-76); male 

75%) and decompensated (DC, n=22; 58 yrs (56-63); M 59%) 

cirrhosis and Acute on Chronic Liver Failure (ACLF, n=5; 60 

yrs (43-67), M 80%) ) and Healthy Controls (HC, n=19; 38 yrs 

(33-41); M 47%). MELD score was 8 (7-11) in SC, 14 (10-27) 

in DC and 38 (23-40) in ACLF. Assays included plasma von 

Willebrand Factor (VWF), its regulator ADAMTS13, Thrombin 

generation in the presence and absence of thrombomodulin 

(TM) to activate protein C, and clot lysis time to test fibrinolytic 

capacity. Results As compared to HC, VWF was elevated in 

all patient groups, with levels increasing with disease severity: 

SC 247% [124-467], p=ns, DC 591% [389-842], P


742 

External validation of risk-stratification criteria in variceal 

bleeding (ChildC-C1, MELD19) for the optimization 

of the use of early TIPS 

Salvador Augustin 1 , Juan Abraldes 2 , Lucio Amitrano 3 , Alba 

Cachero 4 , Maria Anna Guardascione 3 , Jose Castellote 4 , Puneeta 

Tandon 2 , Joan Genescà 1 ; 1 Hospital Universitari Vall d’Hebron, 

Barcelona, Spain; 2 University of Alberta, Edmonton, AB, Canada; 

3 Ospedale Cardarelli, Naples, Italy; 4 Hospital Bellvitge, Barcelona, 

Spain 

Early-TIPS is currently considered the treatment of choice for 

high-risk cirrhotic patients with acute variceal bleeding (AVB). 

However, recent studies have shown that the criteria used to 

define high-risk should be refined. Several alternatives to early-TIPS 

criteria (Child B + active bleeding and Child C) have 

been proposed (Child C + creatinine >1 mg/dL, ChildCC1; 

and MELD ≥19). The aim of the present study was to evaluate 

the external validity of three classification rules (early-TIPS trial 

criteria, ChildCC1 and MELD19) to identify high-risk patients 

eligible for early-TIPS after an ABV. In the present observational, 

retrospective, multicentric, international study, we 

reviewed already collected data from 915 patients with AVB 

from 4 centers which have previously published studies on the 

issue (Naples-Italy; Alberta-Canada; Bellvitge and Vall d’Hebron-Spain). 

All patients were treated with current standard of 

care (drugs + ligation + antibiotics, with TIPS as rescue therapy). 

Most patients had viral (39%) or alcohol (35%) cirrhosis 

(12% had both). Median age was 59 (range 23-90), 64% 

were male. 6-week mortality in the whole cohort was 18%. 

Excluding patients non-eligible for TIPS (age ≥75, creatinine 

≥3 mg/dL, Child score ≥14, HCC beyond Milano criteria or 

portal thrombosis, N=673) mortality was 15%. Mortality for 

Child class and each classification rule for these patients are 

shown in the table. All 3 rules discriminated well high vs. lowrisk 

patients. However, mortality for Child B was significantly 

lower than for Child C patients, regardless of active bleeding. 

Within Child C, mortality was overall high but significantly 

different depending on baseline creatinine (23% in


The following authors have nothing to disclose: Nicole Loo, Khurram Bari, Salvador 

Augustin, Maria Ciarleglio, Yanhong Deng, Mario Strazzabosco 

744 

Inter-observer variability is high for the assessment of 

active bleeding at endoscopy in cirrhotic patients with 

variceal bleeding 

Simona Tripon 1 , Marika Rudler 1 , Maxime Mallet 1 , Christophe 

Bureau 2 , Dominique Thabut 1 ; 1 Hepatology ICU, AP-HP, hôpital 

Pitié-Salpêtrière, Paris, France; 2 Service d’Hépato-Gastroentérologie, 

Hôpital Purpan, Toulouse, France 

Background/Aim: Guidelines stipulate that an early-TIPS 

placement must be considered in patients with Child-Pugh B 

(CPB) cirrhosis and variceal bleeding (VB) with active bleeding 

at initial endoscopy. However, there are no strong data 

confirming that active bleeding is predictive of bad survival 

in CPB patients. Moreover, it is a subjective criterion, which 

inter-observer variability has never been evaluated. The aim 

of this study was to assess the inter-observer variability of the 

endoscopic finding of active bleeding in patients with cirrhosis 

and portal hypertension-related gastrointestinal bleeding 

(GIB), using video-recording of endoscopies. Methods: We 

prospectively included all consecutive patients with cirrhosis 

and upper GIB admitted in Hepatological ICU between October 

2014 and May 2015. Endoscopies were performed by 

a senior hepatologist in charge of the patient. Patients were 

treated according to Baveno V recommendations. Endoscopies 

were recorded and analyzed by 4 other senior hepatologists 

who were blinded to patients’ diagnosis and outcomes 

and to each other findings. Interobserver agreement for endoscopic 

findings was determined using an agreement rate and 

Kappa statistics. Results: Overall, 224 evaluations were performed 

in 56 patients with upper GIB and cirrhosis Patients 

characteristics: male gender: 66.1%; age 60 ± 11.9 yrs ; 

etiology of cirrhosis: alcohol= 52%, viral=20%, other=28% 

; Child-Pugh A/B/C= 12./36/52% ; MELD score 18.8 ± 7 

; shock at admission 23%, source of bleeding according to 

physician in charge of the patient: oesophageal varices 50 %, 

cardial varices 11 %, ulcer 5 %, portal gastropathy 5%, other 

29%). Interobserver agreement for assessment of the source 

of bleeding was excellent (kappa=0.76, CI95%: 0.54-0.88) 

whereas interobserver agreement for the diagnosis of active 

bleeding at endoscopy was poor (kappa=0.55, CI95%: 0.35- 

0.70). Early-TIPS was indicated in 6 CPB patients because of 

active bleeding at endoscopy. Interobserver agreement was 

also poor for the diagnosis of active bleeding at endoscopy in 

those patients (kappa=0.45, CI95%: 0.24-0.60). Conclusion: 

Inter-observer variability in the evaluation of active bleeding at 

endoscopy is very high in patients with cirrhosis and variceal 

bleeding. This criterion should not be used anymore to indicate 

early-TIPS placement. 

Disclosures: 

Marika Rudler - Speaking and Teaching: Gilead, Mayoly 


Gore 

The following authors have nothing to disclose: Simona Tripon, Maxime Mallet, 

Dominique Thabut 

745 

Prevention of Recurrent Variceal Hemorrhage: Individual 

Patient Data (IPD) Meta-Analyses Of Outcomes With 

Current Therapy Based On Severity Of Cirrhosis 

Agustin Albillos 1 , Javier M. González 1 , David Arroyo-Manzano 2 , 

Javier Zamora 2 , Irfan Ahmad 3 , Joaquin de la Peña 4 , Juan Carlos 

Garcia-Pagan 5 , Gin-Ho Lo 6 , Shiv K. Sarin 7 , Jaime Bosch 5 , Juan 

Abraldes 8 , Guadalupe Garcia-Tsao 9 ; 1 Hospital Universitario 

Ramon y Cajal, IRYCIS, CIBEREHD, Madrid, Spain; 2 Hospital 

Universitario Ramon y Cajal, IRYCIS, CIBERESP, Madrid, Spain; 

3 Sheikh Zayed Medical College/Hospital, Rahim Yar Khan, Pakistan; 

4 Hospital Universitario Marqués de Valdecilla, Santander, 

Spain; 5 Hospital Clinic, IDIBAPS, CIBEREHD, University of Barcelona, 

Barcelona, Spain; 6 E-DA Hospital, Kaohsiung, Taiwan; 

7 Institute of Liver and Biliary Sciences, New Delhi, India; 8 University 

of Alberta, Alberta, AB, Canada; 9 Yale University School of 

Medicine, New Haven, CT 

Current standard of care to prevent variceal rebleeding, i.e. 

endoscopic variceal ligation plus beta-blockers (EVL+BB), is 

recommended for all patients. Since risk stratification is essential 

in individualizing care, we aimed to investigate whether 

outcomes of this therapy would differ depending on cirrhosis 

severity. Because data were unavailable from published studies, 

we performed meta-analyses using IPD from fully published 

RCT that compared EVL+BB vs. each therapy alone (BB or EVL) 

and that reported results on all-source rebleeding and mortality 

in patients with cirrhosis who had recovered from acute 

variceal hemorrhage. We performed two meta-analyses: one 

using IPD from 3 trials (389 patients) comparing EVL+BB vs. 

BB (+nitrates if tolerated) (Ahmad 2009, Garcia-Pagan 2009, 

Lo 2009) and the second using IPD from 4 trials (409 patients) 

comparing EVL+BB vs. EVL alone (Ahmad 2009, De la Peña 

2005, Kumar 2009 and Lo 2000). We excluded 4 trials: 2 

abstracts, 1 with HVPG-guided drug therapy, and 1 not reporting 

mortality. Multilevel logistic regression models were fitted to 

perform one-step IPD meta-analyses, with rebleeding and mortality 

as dependent variables and severity of cirrhosis (Child 

A vs. B/C, ascites, bilirubin, albumin) and treatment modality 

as independent variables. Interaction terms between treatment 

modality and covariates were included in the model to test if 

treatment effect varied between subgroups. Results: The first 

meta-analysis (BB control group) failed to show superiority of 

combination therapy vs. BB alone, with a trend for a significant 

benefit in Child A patients. The second meta-analysis 

(EVL control group) shows that combination therapy EVL+BB is 

superior to EVL alone as it reduces rebleeding in all patients, 

with a significant reduction in mortality, particularly in Child 

B/C pts. Conclusions: Results suggest that BB alone are equivalent 

to combination therapy, however these results should be 

taken cautiously given large confidence intervals. In contrast, 

results clearly show that EVL alone carries an increased risk of 

rebleeding and death, indicating that BB are the cornerstone 

of combination therapy in the prevention of recurrent variceal 

hemorrhage.


Disclosures: 

Juan Carlos Garcia-Pagan - Consulting: Novartis; Grant/Research Support: 

GORE 



Guadalupe Garcia-Tsao - Advisory Committees or Review Panels: Abbvie, Fibrogen 

The following authors have nothing to disclose: Agustin Albillos, Javier M. 

González, David Arroyo-Manzano, Javier Zamora, Irfan Ahmad, Joaquin de la 

Peña, Gin-Ho Lo, Shiv K. Sarin, Juan Abraldes 

746 

Serum lysyl oxidase-like-2 (sLOXL2) is correlated with 

the hepatic venous pressure gradient (HVPG) in patients 

with cirrhosis due to hepatitis C 

Nezam H. Afdhal 1 , Gregory T. Everson 2 , José Luís Calleja 3 , Geoff 

McCaughan 4 , Tarik Asselah 5 , Robert P. Myers 6 , Raul E. Aguilar 

Schall 6 , Jill M. Denning 6 , Diana M. Brainard 6 , Mani Subramanian 

6 , John G. McHutchison 6 , Michael R. Charlton 7 , Edward J. 

Gane 8 , Xavier Forns 9 , Jaime Bosch 9 ; 1 Beth Israel Deaconess Medical 

Center and Harvard Medical School, Boston, MA; 2 University 

of Colorado Denver, Aurora, CO; 3 Hospital Puerta de Hierro, 

Madrid, Spain; 4 Royal Prince Alfred Hospital, University of Sydney, 

Sydney, NSW, Australia; 5 Service d’Hépatologie, Hôpital 

Beaujon, Clichy, France; 6 Gilead Sciences, Inc., Foster City, 

CA; 7 Intermountain Medical Center, Murray, UT; 8 University of 

Auckland, Auckland, New Zealand; 9 Liver Unit, Hospital Clinic, 

IDIBAPS and CIBEREHD, Barcelona, Spain 

Background: Noninvasive predictors of clinically significant 

portal hypertension (CSPH) are limited. Our aim was to determine 

the association between sLOXL2, an enzyme involved in 

collagen cross-linkage, and the HVPG in patients with portal 

hypertension due to hepatitis C virus (HCV)-related cirrhosis. 

Methods: The study included patients with portal hypertension 

due to HCV-related cirrhosis who achieved a sustained virologic 

response (SVR12) to a 48-week course of sofosbuvir (SOF) and 

ribavirin (RBV) in study GS-US-334-0125. HVPG was measured 

according to a standardized protocol with centralized 

review at baseline and the end of treatment (EOT). A hemodynamic 

response was defined as a reduction in HVPG to


fixed effects model to assess the primary outcome (incidence 

of gastric variceal rebleeding) and secondary outcomes (complete 

obliteration of gastric varices, aggravation of esophageal 

varices and mortality rate). Review Manager 5.3 software program 

was utilized for statistical analysis. Results: The search 

yielded three studies involving 206 patients. The mean percentage 

of patients with gastric variceal rebleeding was 21% 

in the EIT group and 4% in the BRTO group, with a mean odds 

ratio of 0.13, P < 0.01 (95% CI: 0.03-0.47).The percentage of 

patients with complete obliteration of gastric varices was 74% 

in the EIT group and 95% in the BRTO group: odds ratio 9.40, 

P < 0.01 (95% CI: 2.32-38.02). There was no significant difference 

for aggravation of esophageal varices and mortality 

rate between the two groups. Conclusions: Balloon-occluded 

retrograde transvenous obliteration was more effective than 

endoscopic injection therapy in the eradication of GV and 

prevention of rebleeding. 

Rebleeding rate BRTO vs EIT 

(6.1% vs 6.5%, p=1.000) and 5-day mortality (5.3% vs 4.9%, 

p=1.000) in PPI group and control group. PPI group showed 

less 6-week rebleeding rate (12.1 % vs 22.8%, p=0.031) than 

control group while 6-week mortality was similar between 

two groups (7.6% vs 6.5% respectively, p=0.810). PPI group 

showed better event free survival compared to control group 

over control group (p=0.019 by Log Rank test). Multivariate 

analysis identified Child-Pugh score as a predictor for 5-day 

treatment failure (HR 1.648, 95%CI 1.139-2.384, p=0.008). 

Child-Pugh score and PPI therapy were identified as independent 

predictors for 6-week treatment failure (HR, 1.328, 95%CI 

1.096-1.610 p=0.004 and HR, 0.465, 95%CI 0.233-0.931, 

p=0.031). The occurrence of serious infection was similar 

between two groups (3.8% and 6.5%, respectively p=0.399) 

Conclusion: PPI therapy can offer advantage of decrease the 

rate of recurrent bleeding in cirrhotic patients with acute variceal 

bleeding. PPI could be safely used in patients with liver 

cirrhosis by careful selection of candidate. 

Cox’s univariate and multivariate analysis of predictors of 6-week 

treatment failure 

Disclosures: 

The following authors have nothing to disclose: Dina Ahmad, Mohammad 

Esmadi, Shadi Hamdeh, Kunut Kijsirichareanchai, Chijioke U. Enweluzo, Saurabh 

Kapur, Marco A. Olivera-Martinez 

749 

Prospective validation of the efficacy of proton pump 

inhibitors in acute gastroesophageal variceal bleeding 

in patients with cirrhosis: A single center case-control 

study 

Soo Young Park 1 , Se Young Jang 1 , Su Hyun Lee 1 , Yu Rim Lee 1 , Sun 

Kyung Jang 1 , Won Young Tak 1 , Young Oh Kweon 1 , Jeong Heo 2 , 

Hyun Young Woo 2 , Won Lim 2 , Jung Gil Park 3 , Keun Hur 4 , Gyeonghwa 

Kim 4 , Yong-Hun Choi 4 ; 1 Hepatology, Kyungpook National 

University Hospital, Daegu, Korea (the Republic of); 2 Department 

of Internal Medicine, Pusan National University Hospital, Pusan, 

Korea (the Republic of); 3 Internal Medicine, CHA University Gumi 

CHA Medical Center, Gumi, Korea (the Republic of); 4 Department 

of Biochemistry, Kyungpook National University, Daegu, Korea 


Background and aims: The efficacy and safety of PPI in patient s 

with acute variceal bleeding have not evaluated yet. Therefore, 

we aimed to determine the efficacy and safety of long-term 

proton pump inhibitors in acute gastroesophageal bleeding 

by reviewing prospective cohort of patients with liver cirrhosis. 

Patients and methods: Patients with acute variceal bleeding 

were prospectively evaluated for 5-day in-hospital failure, 

6-week failure and recurrent bleeding afterwards. A total 

of 353 patients with liver cirrhosis visited for acute variceal 

bleeding from 2008 to 2013. We enrolled 255 patients by 

excluding 47 patients with non-variceal bleeding and 51 with 

HCC with portal vein thrombosis. Patients were prospectively 

evaluated according to previous protocols evaluating treatment 

outcome in acute gastroesophageal variceal bleeding (Hepatology 

2015;61:1033) Five day in-hospital treatment failures 

were determined by Baveno V criteria and rebleeding within 

6 weeks were prospectively evaluated Results: No differences 

were observed regarding baseline characteristics of patients 

with PPI group (n=132) and control group (n=123). There 

were no significant differences in 5-day treatment failure rate 

Disclosures: 



Jeong Heo - Advisory Committees or Review Panels: Abbvie; Grant/Research 

Support: Roche 

The following authors have nothing to disclose: Soo Young Park, Se Young Jang, 

Su Hyun Lee, Yu Rim Lee, Sun Kyung Jang, Young Oh Kweon, Hyun Young Woo, 

Won Lim, Jung Gil Park, Keun Hur, Gyeonghwa Kim, Yong-Hun Choi 

750 

Atorvastatin and propranolol combination cause greater 

reduction of HVPG than propranolol alone in patients of 

cirrhosis with portal hypertension 

Saptarshi Bishnu 1 , Avik Sarkar 2 , Saswata Chatterjee 1 , Sk Mahiuddin 

Ahammed 1 , Kshaunish Das 3 , Kausik Das 1 , Gopal K. Dhali 3 , 

Abhijit Chowdhury 1 ; 1 Department of Hepatology, School of Digestive 

and Liver Diseases, Institute of Postgraduate Medical Education 

and Research, Kolkata, India; 2 Department of GI Radiology, 

School of Digestive and Liver Diseases, Institute of Postgraduate 

Medical Education and Research, Kolkata, India; 3 Department of 

Gastroenterology, School of Digestive and Liver Diseases, Institute 

of Postgraduate Medical Education and Research, Calcutta, India 

Introduction:Statins can modulate portal microvascular dynamics 

in cirrhotic patients. We present preliminary data from a 

study aimed at comparing combination of propranolol (Ppnl) 

and atorvastatin versus Ppnl alone in reducing portal pressure 

in cirrhosis. Methods:In this open-label study, 23 consecutive 

cirrhotic patients were randomized into Arm A (atorvastatin 

20 mg daily with Ppnl in incremental dose, n=11) or Arm B 

(incremental dose Ppnl, n=12). Hepatic venous pressure gradient 

(HVPG) was estimated at baseline and after 30 days. 

Results:The 2 arms were matched with respect to etiology, 

Child status and baseline HVPG (Table 1). Decrease of wedged 

hepatic venous pressure, free hepatic venous pressure and 

HVPG in Arm A and Arm B after 30 days were 6.09±3.56 vs 

4.67±2.57 (p=0.290), 1.27±1.67 vs 1.83±2.62 (p=0.546) 

and 4.81±2.82 vs 2.58±1.88 mm Hg (p=0.041) respectively. 

Conclusion:HVPG decrease in cirrhotics treated with atorvasta-


tin and Ppnl is significantly more than those treated with only 

Ppnl. Atorvastatin, with its pleiotropic effects, may be useful in 

portal hypertension in cirrhosis. Larger datasets are required 

for ratification. 

Table 1: Baseline characteristics 

criteria and 81 patients were excluded later because of history 

of EVs endoscopic treatment. Finally, 1269 patients were analyzed. 

The prevalence of EVs/large EVs is 43.4/25.5% in all 

patients and is 32.0/15.4%, 69.3/47.7%, 81.6/61.8% in 

child A, B, C patients, respectively. Platelet


serelaxin was well tolerated. These results support further evaluation 

of the effect of serelaxin on HVPG in patients with PHT. 

Percent reductions in PPG and PVP from baseline during serelaxin 

infusion 

All values are median values. TGT – Thromboplastin generation 

time done using patients adsorbed plasma with control serum and 

control platelets. K – kinetic time, R – Reaction time. 

Disclosures: 

The following authors have nothing to disclose: Vaibhav S. Somani, Apurva 

Shah, Deepak N. Amarapurkar 

753 

Serelaxin reduced portal pressure gradient and portal 

vein pressure in patients with cirrhosis and portal 

hypertension 

Neil J. Lachlan 1 , Neil Masson 2 , Hamish Ireland 2 , Graeme Weir 2 , 

Christopher Hay 2 , Thomas Severin 3 , Neelesh Dongre 3 , Rajnish 

Saini 4 , Judy Pak 4 , David Carr 3 , Denise Yates 5 , John P. Iredale 6 , 

Peter C. Hayes 1 , Jonathan A. Fallowfield 1,6 ; 1 Liver Unit, Royal 

Infirmary of Edinburgh, Edinburgh, United Kingdom; 2 Department 

of Radiology, Royal Infirmary of Edinburgh, Edinburgh, United 

Kingdom; 3 Novartis Pharma AG, Basel, Switzerland; 4 Novartis 

Pharmaceuticals Corporation, East Hanover, NJ; 5 Novartis Institutes 

for Biomedical Research, Cambridge, MA; 6 MRC Centre for 

Inflammation Research, University of Edinburgh, Edinburgh, United 

Kingdom 

Background: Portal hypertension (PHT) accounts for the most 

serious complications of cirrhosis. A reduction in the hepatic 

venous pressure gradient (HVPG) to 20% has 

been reported to improve clinical outcomes, but current drug 

therapy is ineffective in a significant proportion of patients and 

adverse effects (AEs) are common. We have recently shown 

that serelaxin, a recombinant form of the human peptide hormone 

relaxin-2, has anti-fibrotic and portal hypotensive effects 

in cirrhotic rats. Direct measurement of portal hemodynamics 

in response to vasoactive drugs can be performed in patients 

with a transjugular intrahepatic portosystemic shunt (TIPSS) at 

the time of portography assessment. In a two-part exploratory 

open-label study, Part B examined the safety, tolerability, and 

effects of serelaxin on portal and systemic hemodynamics in 

patients with cirrhosis, PHT and a TIPSS. Methods: Patients 

with alcohol-related cirrhosis, a functioning TIPSS confirmed by 

portography, and a portal pressure gradient (PPG = portal vein 

pressure (PVP) - inferior vena cava pressure (IVCP)) >5mmHg 

were treated with serelaxin (i.v. for 60 minutes at 80ug/kg/ 

day then at least 60 minutes at 30ug/kg/day). PPG was measured 

at baseline and post infusion. PVP and blood pressure 

(BP) were measured every 15 and 10 minutes, respectively. 

Safety, including AEs, was assessed. Results: 3 male and 3 

female patients entered the study (mean age 42.8±7.7 years). 

The mean baseline PPG was 8.2mmHg (95% CI 6.1, 11.1). 

Following at least 120 minutes of serelaxin infusion there was 

a 31.3% (95% CI -66.5, 71.6) reduction in the PPG compared 

to baseline. During the infusion there was a progressive reduction 

in the PVP reaching a decrease of 25.2% (95% CI -12.7, 

50.3) from baseline at the 120 minute time point (Table). The 

reduction in PVP started at 30 minutes and continued through 

to the 135 minute time point. With serelaxin infusion, there 

were no newly occurring liver enzyme abnormalities, no clinically 

significant changes in BP, and no discontinuations due 

to AEs. Conclusions: In this small exploratory study serelaxin 

induced a rapid and potentially clinically significant reduction 

in portal pressure. Despite advanced cirrhosis in these patients, 

Disclosures: 

Thomas Severin - Employment: Novartis Pharma AG; Stock Shareholder: Novartis 

Pharma AG 

Neelesh Dongre - Employment: Novartis Pharma AG; Stock Shareholder: Novartis 

Pharma AG 

Rajnish Saini - Employment: Novartis Pharmaceutical Corporation 

Judy Pak - Employment: Novartis Pharmaceuticals 

Denise Yates - Employment: Novartis 

Peter C. Hayes - Advisory Committees or Review Panels: Roche, MSD, Jannsen, 

Gilead, ?ONO, Norgine; Grant/Research Support: Novartis; Speaking and 

Teaching: Roche, MSD, Pfiser, Gore, Falk, Ferring 

The following authors have nothing to disclose: Neil J. Lachlan, Neil Masson, 

Hamish Ireland, Graeme Weir, Christopher Hay, David Carr, John P. Iredale, 

Jonathan A. Fallowfield 

754 

Transjugular intrahepatic portosystemic shunt for portal 

vein thrombosis with variceal bleeding in liver cirrhosis: 

Outcomes and predictors in a prospective cohort study 

Xingshun Qi, Chuangye He, Wengang Guo, Zhanxin Yin, Jianhong 

Wang, Zhengyu Wang, Jing Niu, Ming Bai, Zhiping Yang, 

Daiming Fan, Guohong Han; Xijing Hospital of Digestive Diseases, 

Xijing Hospital, Fourth Military Medical University, Xi’an, China 

This prospective cohort study aimed to assess the risk factors 

associated with transjugular intrahepatic portosystemic shunt 

(TIPS) technical success, outcome, and prognosis in cirrhotic 

patients with portal vein thrombosis (PVT) and a history of variceal 

bleeding. Between May 2009 and April 2011, 51 cirrhotic 

patients with PVT who attempted TIPS procedures for the 

prevention of variceal rebleeding were enrolled. TIPS success 

rate was 84% (43/51). An increased degree of thrombosis 

within the portal trunk and portal vein branches was inversely 

associated with TIPS success. Median follow-up time was 40.07 

months (range: 0.02-56.87). The cumulative risk of rebleeding 

was significantly different between TIPS success and failure 

group (p=0.002). The univariate analysis also demonstrated 

that TIPS failure was the only significant predictor associated 

with rebleeding (hazard ratio [HR]=4.174, 95% confidence 

interval [CI]: 1.558-11.186). In TIPS success group, the cumulative 

rates free of shunt dysfunction at the 6th and 12th month 

were 79% and 76%, respectively. Absence of total superior 

mesenteric vein (SMV) thrombosis was the only independent 

predictor (HR=0.189, 95%CI: 0.047-0.755). In TIPS success 

group, the 1- and 3-year cumulative survival rates were 77% 

and 62%, respectively. Albumin level was the only independent 

predictor (HR=0.877, 95% CI: 0.779-0.986). Successful 

TIPS insertions could effectively prevent from rebleeding in 

cirrhotic patients with PVT and variceal bleeding. Degree of 

PVT and SMV thrombosis was associated with TIPS failure and 

shunt dysfunction, respectively. 

Disclosures: 

The following authors have nothing to disclose: Xingshun Qi, Chuangye He, 

Wengang Guo, Zhanxin Yin, Jianhong Wang, Zhengyu Wang, Jing Niu, Ming 

Bai, Zhiping Yang, Daiming Fan, Guohong Han


755 

Transient Elastography and simple serological tests 

as screening tools for esophageal varices in cirrhotic 

patients 

Simona Bota 1 , Mattias Mandorfer 1 , Remy Schwarzer 1 , Vlad F. 

Iovanescu 2 , Philipp Schwabl 1 , Thomas Reiberger 1 , Michael 

Trauner 1 , Arnulf Ferlitsch 1 , Markus Peck-Radosavljevic 1 ; 1 Gastroenterology 

and Hepatology, Medical University of Vienna, Vienna, 

Austria; 2 Gastroenterology and Hepatology, University of Medicine 

and Pharmacy, Craiova, Romania 

Background:Diagnosis of esophageal varices (EV) in cirrhotic 

patients is crucial for timely initiation of prophylaxis of variceal 

bleeding but requires upper GI endoscopy. Recently,the Baveno 

VI consensus proposed the use of Transient Elastography 

(TE) for exclusion of EV for a value under 20kPa. Our aim 

was to assess the diagnostic value of serological scores (initially 

developed for fibrosis evaluation) and transient elastrography(TE) 

as screening tool for exclusion of EV. Methods:Our 

study included 742 consecutive cirrhotic patients evaluated in 

our Hemodynamic Labor undergoing upper GI endoscopy, 

HVPG measurements, TE and serological tests between 2007 

and 2014. Simple serological scores were calculated: Fibrosis 

Index(FI), FIB-4, Forns score and Lok score. According to the 

newly proposed TE quality criteria, ten valid measurements with 

a median liver stiffness value≥7.1kPa and IQR/median>30% 

were considered poorly reliable and not included into the 

analysis. TE cases without 10 valid measurements were also 

excluded. For TE was used the cut-off proposed by Baveno 

VI consensus und for the other test the cut-offs with the best 

negative predictive value. Results: The etiology of liver cirrhosis 

was: viral-340 patients (45.8%), alcoholic-250 (33.7%), 

others etiologies-152 (20.5%). The incidence of EV in different 

etiologies of liver cirrhosis was: viral-153/340 patients (45%), 

alcoholic-198/250 (79.2%), others-90/152 (59.2%). The proportion 

of Child-Pugh A, B and C patients in each cohort of 

patients was: viral etiology-60%, 30%, 10%; alcoholic etiology 

-31.6%, 37.6%, 30.8% and others etiology -32.9%, 44.1%, 

23%. For a cut-off value


757 

Acute kidney injury predicts mortality in cirrhotic 

patients with gastric variceal bleeding 

Yun-Cheng Hsieh, Kuei-Chuan Lee, Ping-Hsien Chen, Chien-Wei 

Su, Ming-Chih Hou, Han-Chieh Lin; Taipei Veterans General hospital, 

Taipei, Taiwan 

Backgrounds and Aims: Acute kidney injury (AKI) is an important 

complication in patients with cirrhosis. However, the role 

of AKI in patients with gastric variceal (GV) bleeding remained 

unknown. Recently, the international club of ascites (ICA) proposed 

the new definition of AKI in cirrhotic patients. We aim at 

evaluating the ICA criteria and their association with the prognosis 

of cirrhotic patients with GV bleeding. Methods: From 

January 2005 to August 2011, 103 cirrhotic patients admitted 

to Taipei Veterans General hospital due to acute GV bleeding 

were analyzed retrospectively. Results: Forty-one (39.8%) 

patients fulfilled the ICA criteria of AKI, 26 patients (25.2%) in 

stage1, 13 patients (12.6%) in stage 2 and 2 patients (1.9%) 

in stage3, respectively. The severity of liver disease (Child-Pugh 

scores or its component, and MELD scores), shock at admission, 

hematemesis, increased transfusion blood units, elevated 

CRP and the development of infection were associated with the 

occurrence of AKI. Among patients with AKI stage 1 and 2, 8 

(20.5%) progressed to a higher AKI stage. The patients with 

progressed AKI stages had higher child-pugh scores, lower 

serum sodium and higher CRP level. The overall 6-week and 

3-month mortality were 15.5% (n=16) and 20.4% (n=21), 

respectively. In multivariate analysis, the occurrence of AKI 

was significantly associated with a higher 6-week mortality 

rate (34.1% vs. 3.2%, p=0.049). AKI stages were independent 

predictors of 3-month survival (8.1% in patients without AKI, 

30.1% in stage 1, 53.3% in stage 2+3, p=0.002). Conclusions: 

The occurrence of AKI defined by the ICA criteria is high 

in cirrhotic patients with acute GV bleeding. The presence AKI 

was associated with 6-week mortality and the stages of AKI 

further predict 3-month survival. 

Six-week (A) and 3-month (B) mortality in patients with gastric variceal 

bleeding stratified by acute kidney injury (AKI) 

Disclosures: 

The following authors have nothing to disclose: Yun-Cheng Hsieh, Kuei-Chuan 

Lee, Ping-Hsien Chen, Chien-Wei Su, Ming-Chih Hou, Han-Chieh Lin 

758 

Argon Plasma Coagulation Versus Endoscopic Band 

Ligation for Management of Severe Portal Hypertensive 

Gastropathy 

Mohamed M. El-Saadany; Internal Medicine, Mansoura University, 

Mansoura City, Egypt 

INTRODUCTION: Portal hypertensive gastropathy ( PHG ) 

occurs as a complication of cirrhotic or non-cirrhotic portal 

hypertension. PHG is clinically important since it may cause 

insidious or acute severe blood loss. It is characterized 

endoscopically by presence of gastric mucosa abnormality 

described as mosaic-like pattern of snake skin with or without 

red spots. Argon Plasma Coagulation ( APC ) has been used 

as the first therapeutic endoscopic treatment of severe PHG. 

AIM OF STUDY: In this work, Endoscopic Band Ligation ( EBL 

) of affected gastric mucosa has been evaluated as a new 

modality for the treatment of bleeding PHG in comparison with 

the commonly used APC. PATIENTS and METHOD: This is a 

single center, prospective and randomized study included 18 

patients with post-HCV cirrhosis (100%) presented with upper 

GI bleeding admitted at Mansoura University Emergency Hospital 

form Jan.1 st 2013 to Feb.28 th 2014 and were followed 

up for 6 months after complete eradication of PHG. The 18 

patients were randomized 2:1 into group 1 (APC) including 12 

patients: 7 males and 5 females with age (meana± SD) 58.33 

±9.61years and group 2 (EBL) including 6 patients 1 male and 

5 females with age (mean± SD) 62.67±4.55 years. RESULTS: 

The efficacy to eradicate PHG was 83.3% in APC group and 

100% in EBL group (ρ. > 0.05). The number of endoscopic sessions 

needed to eradicate PHG was significantly (ρ. 0.03) less 

in EBL group (3.17±0.75) than in APC group (4.08±0.79) and 

the duration of endoscopic procedures was also significantely 

( ρ.0.001) shorter in EBL group (275.5±35.5 sec.) than in 

APC group (469.75±60.22 sec.). Adverse events like duration 

of distention was significantly (ρ.0.001) less in EBL group 

(3.0 ±3.29 min.) than in APC group (25.0±12.75 min.) and 

duration of nausea was significantly (0.02) less in EBL group 

(3.0±3.29 min.) than APC group (12.0±8.0 min.). However, 

gastric ulcers were found in 2 patients in EBL group and none 

in APC group (ρ.0.03) while gastric polyps were more common 

in APC group (41.7%) than in EBL group (ρ.0.06). CON- 

CLUSION: There is no significant difference in the efficacy 

to ablate PHG using either APC or EBL. However, EBL could 

achieve early ablation of PHG with less number of endoscopic 

sessions and fewer adverse effects. 

Disclosures: 

The following authors have nothing to disclose: Mohamed M. El-Saadany 

759 

The impact of esophagogastric varices on the prognosis 

of patients with hepatocellular carcinoma 

Chien-Wei Su 1,2 , Ping-Hsien Chen 3,2 , Yi-Hsiang Huang 1,4 , Teh-Ia 

Huo 1,5 , Ming-Chih Hou 3,2 , Han-Chieh Lin 1,2 , Jaw-Ching Wu 4,6 ; 

1 Division of Gastroenterology, Department of Medicine, Taipei 

Veterans General Hospital, Taipei, Taiwan; 2 Faculty of Medicine, 

School of Medicine, National Yang-Ming University, Taipei, Taiwan; 

3 Endoscopy Center for Diagnosis and Treatment, Taipei 

Veterans General Hospital, Taipei, Taiwan; 4 Institute of Clinical 

Medicine, School of Medicine, National Yang-Ming University, 

Taipei, Taiwan; 5 Institute of Pharmacology, National Yang-Ming 

University, Taipei, Taiwan; 6 Division of Translational Research, 

Department of Medical Research, Taipei Veterans General Hospital, 


Background: Esophagogastric varices (EGV) is an important 

prognostic factor for patients with advanced liver disease, but 

whether it could determine the outcomes of patients with hepatocellular 

carcinoma (HCC) is still obscure. Aims: To assess the 

impact of EGV on the prognosis of patients with HCC. Methods: 

We enrolled 990 treatment-naïve HCC patients who also 

received esophagogastroduodenoscopy at the time of HCC 

diagnosis from 2007 to 2012. Results: A total of 480 (48.5%) 

patients had EGV, including 399 patients had esophageal 

varices (EV) alone, 12 had gastric varices (GV) alone, and 

69 had both EV and GV. Compared to those without EGV, 

patients with EGV were younger in age, and had poorer liver


functional reserve, such as higher rates of ascites and hepatic 

encephalopathy, lower platelet counts, lower serum albumin 

levels, higher bilirubin, alanine aminotransferase, aspartate 

aminotransferase (AST), and alkaline phosphatase levels, and 

more prolonged prothrombin time. Besides, patients with EGV 

were more with multinodular lesions, tumor vascular invasion, 

advanced tumor stage, and lower rates for curative treatment. 

After a median follow-up of 13.1 months (25-75 percentiles 

3.6-36.9 months), 533 patients died. The cumulative 5-year 

survival rates were 24.9% and 46.4% in patients with and 

without EGV, respectively (p


majority of patients were treated for splanchnic vein thrombosis 

(12/20) whereas in the TAC group the majority of patients 

were treated for non-splanchnic venous thrombosis (12/19). 

The TAC group had been treated for a mean of 478 days of 

therapy compared to 253 days in the DOAC group (p=0.2). 

In the DOAC group, 9/20 (45%) were exposed to traditional 

anticoagulation prior to initiation of DOAC. There were 3 documented 

bleeding events in the TAC and 4 bleeding events 

in DOAC (p=0.9). There were 2 major bleeding events in the 

TAC group and 1 major bleeding event in the DOAC group. 

There was 1 moderate bleeding event in each group and 2 

mild bleeding events in the DOAC group. On univariate regression 

and Cox proportional hazard modeling there were no significant 

predictors of bleeding, including concurrent antiplatelet 

therapy. Conclusions: DOAC display similar safety characteristics 

compared to TAC in cirrhosis patients. We did not identify 

any characteristic that predicted a bleeding event while on 

anticoagulation. Larger, prospective studies with anticoagulant 

agents are needed to further define safety, pharmacokinetics 

and efficacy in patients with cirrhosis. 

Bleeding events 

ICH- intracranial hemorrhage 

Disclosures: 

Curtis K. Argo - Independent Contractor: Salix Pharmaceuticals 

Patrick Northup - Consulting: Janssen 



NGM 

The following authors have nothing to disclose: Nicolas Intagliata, Zachary 

Henry, Hillary Maitland, Neeral L. Shah 

762 

Transient elastography evaluation of hepatic and spleen 

stiffness in patients with hepatosplenic schistosomiasis 

Zulane D. Veiga 1 , Cristiane A. Villela Nogueira 2 , Renata D. 

Perez 2 , Homero S. Fogacas 2 , Flavia F. Fernandes 1 , Gustavo 

Pereira 1 , João Luiz Pereira 1 , Carlos Eduardo C. Marques 2 , Marta 

Cavalcanti 2 ; 1 Gastroenterologia e Hepatologia, Hospital Federal 

de Bonsucesso, Rio de Janeiro, Brazil; 2 Hepatologia, Hospital Universitário 

Clementino Fraga Filho, Rio de Janeiro, Brazil 

Hepatosplenic schistosomiasis (HES) is characterized by portal 

hypertension and periportal fibrosis. Transient elastography 

(TE) has been used to evaluate liver and spleen stiffness in 

many liver diseases. To our knowledge, schistosomiasis has not 

been evaluated by TE so far, and its correlation with ultrassonographic 

(US) findings related to portal hypertension remains 

to be defined. Objective: To assess liver and spleen stiffness by 

TE in patients with HES in comparison to compensated cirrhotic 

patients and its relation to US findings. Methods: Sixty nine 

patients were prospectively included in the study: 29 had HES, 

23 had HCV-related liver cirrhosis with esophageal varices 

and 17 were healthy controls. Liver and spleen stiffness values 

were assessed using the FibroScan® 502 equipment (Echosens, 

Paris, France) by an experienced operator. Ten successful 

measurements were carried out for each patient. Liver and 

spleen stiffness values were considered adequate if the success 

rate were at least 60% and the interquartile range (IQR) were 

< 30% of the median value. TE range values are 3.5 to 75 kPa. 

Patients with liver stiffness (LS) values > 9.5 Kpa were classified 

as having significant liver stiffness and spleen stiffness (SS) = 

75 kPa as high spleen stiffness. Patients with HES were submitted 

to ultrasound with dopplerfluxometry (Doppler Hitachi 

EUB 5500 3,5 MHz probe) by a well-trained examinator in 

World Health Organization (WHO) protocol for US assessment 

of schistosomiasis. Values are expressed as percentage 

and mean±SD. Results: Patients with HES had LS significantly 

higher than controls and lower than cirrhotic patients: 16.3 ± 

19,9 vs 3,7 ± 0,7 vs 29.7±15,4 kPa (p= 0,001). SS values 

were comparable between HES and cirrhotic patients: 57.5 

±18,5 vs 54 ± 21,6 kPa (p=0,4) and were significantly higher 

than controls (p= 0,001). Significant LS was observed in 52% 

of HES patients and was associated with higher values of aminotransferases: 

AST 45,7 ± 21 vs 32 ± 9,6 ALT 48 ± 22 vs 

31,7±10,8 (P< 0,05). SS values were = 75 kPa in 43% of 

HES patients while 24% of cirrhotic and no controls presented 

this value (p=0,06). In HES patients, high SS was associated 

to higher values of splenic artery resistance index (p=0,003), 

portal vein diameter( p=0,09), portal vein congestion index 

(p= 0,05) and splenic diameter (p= 0,09). Conclusion: HES 

patients present a distinct pattern of liver and spleen elastography 

in comparison to cirrhotic patients, with lower frequency 

of significant LS and higher frequency of high SS. In HES population, 

elevated spleen elastography may be a surrogate of 

splenic artery resistance and portal vein congestion. 

Disclosures: 

The following authors have nothing to disclose: Zulane D. Veiga, Cristiane A. Villela 

Nogueira, Renata D. Perez, Homero S. Fogacas, Flavia F. Fernandes, Gustavo 

Pereira, João Luiz Pereira, Carlos Eduardo C. Marques, Marta Cavalcanti 

763 

Shunt or meso-portal bypass for portal hypertension in 

children 

Neslihan Celik, Geoffrey Bond, Kyle A. Soltys, Rakesh Sindhi, 

George V. Mazariegos; Hillman Center for Pediatric Transplantation, 

Children’s Hospital of Pittsburgh of UPMC, Pittsburgh, PA 

Purpose To review surgical porto-systemic or meso-portal 

bypass techniques and post-surgical short and long term outcomes 

for the children with portal hypertension (PH). Methods 

Children who underwent porto-systemic or meso-portal bypass 

surgery for PH were reviewed retrospectively in terms of underlying 

or concomitant pathology, patient characteristics, surgical 

techniques, radiologic patency and clinical outcomes. Results 

Between 2002 and 2014, 40 patients underwent 46 bypass 

surgeries consisting 26 spleno-renal shunt (SRS), 9 meso-caval 

shunt (MCS), 8 meso-Rex bypass (MRB), 1 MRB-SRS and 

1 spleno-adrenal shunt. 34 patients with long term follow up 

(median 2.85 year, range 0.49-13.28 years) were analyzed. 

Extra-hepatic portal vein obstruction (EHPVO) and variceal 

bleeding were the indications for surgery. Porto-systemic shunts 

were done when MRB was not technically feasible. There was 

underlying liver disease in 17 patients (Group-A) and normal 

liver parenchyma with EHPVO in 17 patients (Group-B). The 

liver pathologies in Group-A were post-liver transplantation 

portal vein thrombosis (n=6), idiopathic liver fibrosis (n=5), 

total parenteral nutrition induced liver fibrosis (n=3), biliary 

atresia (n=2) and primary sclerosing cholangitis (n=1). Thrombophilia 

(n=6) and umbilical vein catheterization (n=5) were 

the concomitant pathologies in Group-B with idiopathic subgroup 

(n=6) presenting no other abnormal finding. Median 

patient age was 6.65 years (range 0.65-18.55 years). Three 

patients had re-shunt surgery due to shunt thrombosis. Three 

patients underwent liver transplantation because of end stage 

liver disease (n=2) and porto-pulmonary hypertension (n=1) in 

Group-A. One syndromic patient with thrombophilia died 5


years after shunt surgery because of pneumonia in Group B. 

The patient outcomes and comparison between 2 groups were 

listed in the Table below: Conclusion Extra-hepatic 

portal vein obstruction with or without underlying liver disease 

is the major cause of portal hypertension in children. Both porto-systemic 

and meso-portal bypass surgeries were very successful 

in the control of variceal bleeding demonstrated long 

term radiologic patency and clinical outcomes. 

Conclusions: Our data show that using proposed cut-offs of 

LSM and platelet count to rule out the presence of clinically significant 

varices is problematic. A LSM


Abdullah M. Al-Osaimi - Advisory Committees or Review Panels: Abbvie, Gilead, 

Intercept Pharmaceuticals; Grant/Research Support: Chronic Liver Disease 

Foundation (CLDF), Beckman Coulter Inc., Conatus Pharmaceuticals, BioPharma 

Alliance, Bayer HealthCare Pharmaceuticals, Inc., Vital Therapies Inc., Roche 

Molecular Systems, Inc., Abbvie 

The following authors have nothing to disclose: Paul Chang, Daniel Baik 

766 

Comparison of Beta blockers effects on All-Cause Mortality 

in Gastric Varices in an Urban Population 

Paul Chang, Daniel Baik, James Langworthy, Jeffrey Barry, Abdullah 

M. Al-Osaimi; Hepatology, Temple University Hospital, New 

York, NY 

Background: Gastric variceal bleeding although less frequent 

than esophageal variceal bleeding are associated with higher 

mortality and morbidity. Bleeding rates have been reported 

to be 16% at one year, 36% at 3 years and 44% at 5 years. 

There is a relative paucity of data in regards to the best management 

options for these patients. Our aim was to determine 

the effects of beta blockers therapy on patients diagnosed with 

gastric varices on all-cause mortality and sentinel bleeding in 

patients without history of bleeding from gastric varices. Methods: 

A retrospective chart review was conducted on subjects 

identified as having gastric varices from 2011-2013. Subjects 

were identified by our center database (Provation® MD) query. 

Once subjects were identified subjects were followed by chart 

review (EMR) until the last known follow up, additional deaths 

were identified by searching the Social Security database. Initial 

MELD, Child-Pugh, and etiology of cirrhosis were calculated 

at time of diagnosis, as well as heart rate, and use of beta 

blockers. Follow up data included initiation of beta blockers, 

discontinuation of beta blockers, bleeding events, source of 

bleed, and death. Data was analyzed using SAS V9.4. Results: 

A total of 85 patients were identified with no documented 

bleeding from gastric varices, with a mean age of 56.54 ± 

9.6 years, 61 (71.8%) were male, mean MELD was 12.2 ± 10 

with a mean follow up period of 1.82 ± 1.52 years. A total of 

28 patients were on beta blockers at the time of diagnosis. An 

additional 19 patients were started on beta blockers therapy 

after diagnosis, 38 patients received no beta blockers. Figure 

1 shows a Kaplan Meier curve for gastric variceal bleeding 

free survival. In addition, having a heart rate < 80beats/min 

trended towards significance for overall mortality, with 11 

(26%) subjects with heart rate < 80 compared to 18 (43%) subjects 

with heart rate > 80. Conclusion: Beta blockers appear 

to be protective in all-cause mortality in patients with gastric 

varices compared to patients who were not on beta blockers. 

There is further evidence of this as having a heart rate


768 

Effect of nonselective beta-blockade on prehepatic 

portal hypertension evaluated in patients by combined 

spleen pulp and hepatic venous pressure gradient measurements 

Michael Sørensen 1 , Lars P. Larsen 2 , Gerda E. Villadsen 1 , Niels 

Kristian Aagaard 1 , Henning Grønbæk 1 , Peter Ott 1 , Hendrik Vilstrup 

1 , Susanne Keiding 1,3 ; 1 Dept. of Hepatology & Gastroenterology, 

Aarhus University Hospital, Aarhus, Denmark; 2 Dept. of 

Radiology, Aarhus University Hospital, Aarhus, Denmark; 3 Dept. 

of Nuclear Medicine & PET Center, Aarhus University Hospital, 

Aarhus, Denmark 

Prehepatic portal hypertension is a clinically important cause 

of esophageal and gastric varices. β-blockers have an established 

prophylactic role against development and bleeding 

from esophageal and gastric varices in intrahepatic portal 

hypertension but the effect on prehepatic portal hypertension 

is unknown. Here, we evaluated the effect of β-blockade on the 

pressure gradient from spleen pulp to free hepatic vein which 

determines the overall risk of developing varices in prehepatic 

portal hypertension. Material and Methods: Ten patients underwent 

combined measurements of the pressure in the spleen pulp 

and hepatic vein pressure gradient (HVPG) while on and off 

treatment with propranolol (n=9) or carvedilol (n=1). Results: 

The spleen to free hepatic vein pressure gradient decreased 

in eight patients, increased slightly in one patient, and was 

unchanged in one patient. The mean gradient was 29 mm 

Hg without treatment and 24 mm Hg with treatment (P


versus TCS in the management of gastric variceal bleeding. A 

meta-analysis was performed using pooled estimates with odds 

ratio by fixed effects model to assess the primary outcome (incidence 

of gastric variceal rebleeding) and secondary outcomes 

(development of encephalopathy and mortality rate). Review 

Manager 5.3 software program was utilized for statistical analysis. 

Results: Three studies met the inclusion criteria. A total of 

175 patients were included in the meta-analysis. The use of 

TCS for management of gastric variceal bleeding showed a 

statistically significant decrease in the incidence of rebleeding 

(OR 9.19; 95% CI: 1.96-43.05, p


Disclosures: 


Squibb 













The following authors have nothing to disclose: Philipp Schwabl, Theresa Bucsics, 

Remy Schwarzer, Arnulf Ferlitsch 

772 

Does Haemodynamic or Clinical Rebound Exist in 

patients with Cirrhosis after Abrupt Interruption of 

Beta-blockers? 

Audrey Payance 2 , Julien Bissonette 2 , Olivier Roux 2 , Laure Elkrief 2 , 

claire francoz 2 , Ouardia Nekachtali 1 , Dominique C. Valla 1 , Didier 

Lebrec 1 , Francois Durand 2 , Pierre-Emmanuel Rautou 1 ; 1 Paris VII 

University and INSERM, BEAUJON HOSPITAL, Clichy, France; 

2 HEPATOLOGY, BEAUJON HOSPITAL, Clichy, France 

Background and Aims: Beta-blockers (BBs) may be interrupted 

in patients with portal hypertension. The concept of rebound 

after abrupt interruption of BBs is based on animal studies 

showing a transient β-adrenergic hypersensitivity state following 

propranolol withdrawal and on few case reports of 

variceal bleeding following abrupt interruption of this drug. 

However, no human study addressed this issue yet. Our aim 

was thus to determine if a rebound after abrupt interruption of 

BBs exists in patients with cirrhosis both on a hemodynamic 

and a clinical point of view. Patients and Methods: This prospective 

observational study included all patients with cirrhosis 

undergoing right heart and hepatic vein catheterization in our 

hemodynamic laboratory. Four groups of patients were a priori 

defined: “no BBs” defined as patients not receiving BBs; 

“≥ 1day” defined as patients having received BBs within 1 

day before catheterization; “2-3 days”, defined as patients 

having interrupted BBs 2 or 3 days before catheterization; 

and “≥4days” defined as patients having interrupted BBs 4 

days or more before catheterization. Results are presented as 

median (interquartile range). Results: A total of 150 patients 

were included (men, 76%; median age, 57 yrs; alcoholic cirrhosis, 

75%; large varices, 73%). Indications for right heart 

and hepatic vein catheterization included evaluation prior to 

liver transplantation (n=90) or liver surgery (n=8) and assessment 

of cirrhosis severity (n=52). There were 81, 44, 11, 14 

patients in the “no BBs”, “≥1day”, “2-3 days” and “≥4days” 

groups, respectively. MELD and Child-Pugh scores were not 

different among the groups. Hepatic venous pressure gradient 

was not different between patients in groups “≥ 1day” (18.5 

mm Hg, 15-23), “2-3 days” (17 mm Hg, 13-23) and “≥4days” 

(19.5 mm Hg, 17-26). Cardiac index remained low up to 3 

days after BBs interruption. Patients in the “≥4days” group 

had higher cardiac index than those in the “≥1day” group 

[4.6 (3.5-5.1) vs 3.4 (2.6-4.0); p=0.001)] or in the “2-3 days” 

group [4.6 (3.5-5.1) vs 3.1 (2.7-3.7); p=0.007], but only borderline 

significantly higher than patients not receiving BBs [4.6 

(3.5-5.1) vs 3.7 (3.0-4.5); p=0.052]. Among the 25 patients 

in the groups “2-3 days” and “≥4days”, median duration of 

BBs interruption was 4 (3-6) days. No gastrointestinal bleeding 

occurred during that period. Conclusions: Abrupt interruption 

of BBs may slightly increase cardiac index without rebound 

in hepatic venous pressure gradient and with no apparent 

increase in the risk of variceal bleeding. Thus, interruption 

of BBs in cirrhotic patients with portal hypertension does not 

require particular dosing or surveillance. 

Disclosures: 




The following authors have nothing to disclose: Audrey Payance, Julien Bissonette, 

Olivier Roux, Laure Elkrief, claire francoz, Ouardia Nekachtali, Dominique 

C. Valla, Didier Lebrec 

773 

High risk of misclassifying liver stiffness using 2D shearwave 

and transient elastography during a moderate or 

high calorie meal 

Maria Kjærgaard 1 , Maja Thiele 1 , Bjørn S. Madsen 1 , Christian 

Jansen 2 , Jonel Trebicka 2 , Aleksander Krag 1 ; 1 Odense University 

Hospital, Odense, Denmark; 2 University of Bonn, Bonn, Germany 

Introduction: Transient Elastography (TE) (FibroScan) and 

real time 2D shear wave elastography (2D-SWE) (Aixplorer) 

are widely used in the grading and staging of liver fibrosis. 

In patients with chronic viral hepatitis a meal of 600 kcal 

increases liver stiffness (LS) measured by TE. Thus, food intake 

may hamper interpretation of results and clinical decision-making. 

This study aimed to investigate the impact of a meal on 

LS measured by both TE and 2D-SWE in patients with alcoholic 

fibrosis, the role of the meal size and the time it takes 

to return to baseline. Methods: Patients with a liver biopsy 

or known cirrhosis participated in a two-day protocol with a 

250ml/625 kcal liquid meal (49% carbohydrates, 36% fat, 

15% protein) and 500ml/1250 kcal meal on day 1 and 2 

respectively. TE and 2D-SWE were measured at 0, 20, 40, 60, 

120 and 180 minutes. DLS are the highest increase in LS from 

baseline. Results: 51 patients participated: 71% male, median 

age 61years, 88% with alcoholic etiology, 8% HCV and 4% 

NASH. 14 patients showed none or minimal fibrosis (METAVIR 

F0-1), 9 patients moderate or severe fibrosis (F2-3) and 28 

patients cirrhosis (F4). On day 1 and 2, post-meal LS increased 

significantly in all fibrosis groups (see table). The highest DLS 

was measured after a median of 60 minutes (range 20-180) 

and the largest DLS were observed in patients with cirrhosis 

(43% increase). In these patients, LS increased similarly on day 

1 and 2, despite twice the volume and calories (P= 0.93). In 

patients with F0-F3 fibrosis there was a trend towards a higher 

DTE on day 2; F0-1 (P= 0.08) and F2-3 (P=0.07). In patients 

with baseline TE


2D-SWE (P=0.14). On day 2, the same was true for 72% and 

79% (both P18, who underwent a liver biopsy. The 

stage of cirrhosis was identified based on liver biopsy according 

to the Batts-Ludwig fibrosis model. The data was randomly 

separated into two datasets (training 80% and testing 20%) 

after excluding cancer or non-liver disease. A stepwise logistic 

regression model was used to select candidate predictors from 

the variables of routine biomarkers, demographics and behavioral 

risk factors in the training dataset. Then, a novel diagnosis 

index was developed according to the effect of each candidate 

predictor and their correlations. The area under the curve 

of receiver operating characteristic (AUROC) was applied to 

compare the new index to existing ones (Fibro_Q, FIB4, APRI 

AAR), which was also validated in the testing dataset. Results: 

A total of 9 routine biomarkers were used to predict cirrhosis in 

the new index, including: aspartate aminotransferase, alanine 

transaminase, alkaline phosphatase, international normalization 

ratio, total protein, platelet count, blood urea nitrogen, 

albumin, and bilirubin. The new index had significantly higher 

AUROC (0.83, 95%CI 0.79-0.87) than Fibro_Q (0.80, CI 

0.76-0.85), FIB4 (0.79, CI 0.74-0.83), APRI (0.74, CI 0.69- 

0.78), and AAR (0.72, CI 0.67-0.78). Similar results were 

775 

Usefulness of Shear Wave Elastography in the evaluation 

of liver fibrosis in patients with HCV-related liver 

disease: a comparison with Transient Elastography 

Silvia Tonello 2 , Paola Bizzotto 2 , Sara Piovesan 3 , Antonietta 

Romano 2 , Georgios Anastassopoulos 3 , Alfredo Alberti 3 , Marta 

Tonon 2 , Paolo Angeli 2,1 , Giancarlo Bombonato 1 ; 1 Department of 

Medicine DIMED, Unit of Medical Emergencies in Liver Transplantation, 

University of Padova, Padova, Italy; 2 Department of Medicine 

-DIMED, University of Padova, Padova, Italy; 3 Department of 

molecular Medicine, University of Padova, Padova, Italy 

Aim of the study. To evaluate the applicability and performance 

of Shear Wave Elastography (SWE) in the assessment of liver 

fibrosis in healthy volunteers and in patients with HCV-related 

chronic liver disease, in comparison to transient elastography 

(TE). Methods. 48 patients affected by HCV-related chronic 

hepatitis or cirrhosis (19 male and 29 female, mean age 61+ 

11, 25 with a previous diagnosis of chronic hepatitis and 23 of 

compensated cirrhosis) and 8 healthy volunteers (males, mean 

age 30+ 6) underwent SWE with the US machine Toshiba 

Aplio 500 Platinum and TE with Fibroscan (Echosens), on the 

same day, performed by two different operators. TE calculates 

the velocity of propagation through the liver tissue of a low-frequency 

transient shear wave produced by a mechanical probe. 

SWE measures the velocity of propagation of shear waves 

generated by an US induced-radiation force impulse. Toshiba 

Aplio 500 Platinum provides a quantitative color-code elasticity 

image in a box presented on the B-mode image; the liver 

stiffness was obtained from the average of a circular region 

of interest (2 cm in diameter). TE and SWE stiffness values 

were expressed in kPa. A semiquantitative assessment of liver 

steatosis was also performed according to the hyperecogenicity 

and US attenuation of liver parenchyma. Results. Valid 

TE and SWE measurements were obtained in all patients and 

controls. In all healthy subjects TE and SWE measurements 

were within the normal range (TE


sis may lead to an underestimation of the degree of fibrosis by 

TE. Further studies are needed to investigate the correlation of 

SWE measurements with liver fibrosis and steatosis assessed 

with biopsy. 

Disclosures: 

Alfredo Alberti - Advisory Committees or Review Panels: Merck, roche, Gilead, 

Merck, roche, Gilead, Merck, roche, Gilead, Merck, roche, Gilead; Grant/ 

Research Support: Merck, gilead, Merck, gilead, Merck, gilead, Merck, gilead; 

Speaking and Teaching: novartis, BMS, novartis, BMS, novartis, BMS, novartis, 

BMS 

Paolo Angeli - Advisory Committees or Review Panels: Sequana Medical 

The following authors have nothing to disclose: Silvia Tonello, Paola Bizzotto, 

Sara Piovesan, Antonietta Romano, Georgios Anastassopoulos, Marta Tonon, 

Giancarlo Bombonato 

776 

ELF Test Thresholds for disease stratification and prognosis 

in chronic liver disease. 

James W. Day 1 , William M. Rosenberg 1 , Julie Parkes 1,2 ; 1 Institute 

for Liver and Digestive Health, University College London, London, 

United Kingdom; 2 Public Health Sciences and Medical Statistics, 

University of Southampton, Southampton, United Kingdom 

BACKGROUND: Clinically meaningful categories of liver fibrosis 

have traditionally been based on histological staging of 

liver biopsies into mild, moderate and severe fibrosis, and 

cirrhosis. Categorization by disease severity is used to stratify 

patients and carries prognostic significance. Non-invasive 

blood tests for liver fibrosis generate scores that may track 

fibrosis across the categorical stages defined by histology. As 

they are continuous variables they have the potential to provide 

information about fibrosis and prognosis between as well as at 

the thresholds defined by histology. We have investigated the 

performance of the ELF test both at, and between the manufacturer’s 

thresholds and determined the associated prognosis. 

In addition we have identified a more specific threshold for 

cirrhosis. METHODS: A cohort of 1,000 patients with mixed 

chronic liver disease was used to evaluate the performance of 

the manufacturer’s recommended thresholds of ELF score. An 

additional threshold for detecting cirrhosis with 97% specificity 

was derived by analyzing the ROC performance of ELF in this 

cohort. These same thresholds were then used to determine the 

time to first incidence of a liver related event (bleeding varices, 

ascites, encephalopathy, hepatocellular cancer, liver transplantation 

or liver related death) in a subset of 460 patients from 

the cohort followed up for 7 years. RESULTS: Ruling-out fibrosis: 

Fewer than 15% of patients with liver fibrosis >S1/6 will have 

an ELF score 9.8 has a specificity to rule in S5,6 with only a 

10% false positive rate and a score >11.3 has 97% specificity 

for cirrhosis. An increase in the ELF score of 0.8 is associated 

with a clinically significant difference in histological severity 

of fibrosis. Prognosis: The risk of a liver related event within 5 

years was approximately 2%, 30% and 50% respectively for 

patients with ELF scores in the range 7.7-9.79, 9.8-11.29 and 

≥11.3 ELF respectively. CONCLUSIONS: The ELF thresholds 

7.7, and 9.8 stratify patients with CLD into clinically meaningful 

groups for disease severity and prognosis. The ELF test 

threshold of 11.3 improves specificity for cirrhosis and adds 

meaningful prognostic information. The use of a continuous 

variable score to assess liver fibrosis permits more refined stratification 

of fibrosis severity than histological staging. 

Disclosures: 




Julie Parkes - Stock Shareholder: iQur (spouse is shareholder) 

The following authors have nothing to disclose: James W. Day 

777 

Changes in liver stiffness by transient elastography (TE) 

and serum lysyl oxidase-like-2 (sLOXL2) in patients with 

cirrhosis treated with ledipasvir/sofosbuvir (LDV/SOF)- 

based therapy 

Marc Bourlière 1 , Veronique Loustaud-Ratti 2 , Sophie Metivier 3 , 

Vincent Leroy 4 , Armando Abergel 5 , Robert P. Myers 6 , Raul E. 

Aguilar Schall 6 , Robert H. Hyland 6 , Mani Subramanian 6 , John 

G. McHutchison 6 , Lawrence Serfaty 7 , Victor de Ledinghen 8 ; 

1 Hépato-Gastro-Entérologie, Hôpital Saint Joseph, Marseilles, 

France; 2 Recherche Clinique et de l’Innovation, CHU de Limoges 

and Inserm UMR 1092, Université de Limoges, Limoges, France; 

3 Service d’Hépato-Gastro-Entérologie, Hôpital Purpan, Toulouse, 

France; 4 Hépato-Gastro-Entérologie, CHU de Grenboble, Grenoble, 

France; 5 Médecine Digestive, CHU Estaing, Clermont-Ferrand, 

France; 6 Gilead Sciences, Inc., Foster City, CA; 7 Hépatologie, 

Hôpital Saint Antoine, Paris, France; 8 Service d’Hépato-Gastro-Entérologie 

et d’Oncologie Digestive, CHU de Bordeaux, Pessac, 

France 

Background: Liver stiffness measurement (LSM) by TE is routinely 

used to monitor HCV-related fibrosis. Our aim was to 

explore changes in liver stiffness and a novel fibrosis marker, 

sLOXL2, in cirrhotic patients with a sustained virologic response 

(SVR) to SOF/LDV-based therapy. Methods: The study included 

adults with HCV cirrhosis who achieved an SVR in the SIR- 

IUS trial comparing LDV/SOF+ribavirin for 12 weeks vs. LDV/ 

SOF for 24 weeks. All patients had cirrhosis defined by liver 

biopsy, or LSM by TE >12.5 kPa (FibroScan®; Echosens; Paris, 

France), or Fibrotest >0.75 and APRI >2.0. TE was performed 

at baseline (BL) and at the SVR24 time point, and sLOXL2 

was measured by immunoassay (VIDAS® LOXL2; bioMérieux, 

Marcy L’Etoile, France). The estimated stage of fibrosis based 

on TE at BL and SVR24 was categorized as F0-2 (


Estimated fibrosis stage by TE at baseline and SVR24 

Disclosures: 






Squibb 

Veronique Loustaud-Ratti - Board Membership: Gilead, Roche, Schering Plough 

MSD; Speaking and Teaching: Roche, Schering Plough MSD, Janssen, Bristol 

meyers squibb, gilead, Abbvie 

Sophie Metivier - Speaking and Teaching: Roche, BMS, Janssen, Merck, Gilead, 

abbvie 








Sciences, Inc 



Sciences 

Lawrence Serfaty - Board Membership: BMS, Gilead; Consulting: Merck; Speaking 

and Teaching: Roche, Janssen, Merck, Janssen, BMS, Gilead 



778 

A Hepatic Stellate Cell Secreted Protein Signature that 

Has Potential for use in the Non-invasive Detection of 

Fibrosis and Fibrogenic Activity in Chronic Liver Diseases 

Daniel L. Motola 1 , Chan Zhou 1 , Samuel York 1 , Yujin Hoshida 2 , 

Alan Mullen 1 , Raymond T. Chung 1 ; 1 MGH, Boston, MA; 2 Division 

of Liver Diseases,, Icahn School of Medicine at Mount Sinai, New 

York, NY 

Background: Emerging clinical data highlight that the degree 

of hepatic fibrosis correlates with clinical outcomes in patients 

with liver disease (Ekstedt et al Hep. 2015). However, there 

remains no reliable method for detection of fibrosis, other 

than liver biopsy, which carries potential morbidity. Furthermore, 

non-invasive methods such as Fibroscan do not predict 

early stages of fibrosis well. Accordingly, development 

of non-invasive techniques to detect hepatic fibrosis and its 

progression is critically needed. It is well-known that activated 

hepatic stellate cells (HSCs) deposit extracellular collagen 

matrix and directly contribute to fibrosis. Furthermore, studies 

reveal direct correlation between HSC number and degree of 

hepatic fibrosis. Therefore, we hypothesize that HSC-derived 

secreted proteins can be used as functionally relevant, non-invasive 

biomarkers to characterize disease stage and identify 

high-risk patients. Methods: We performed RNA-sequencing 

of primary human activated HSCs to comprehensively characterize 

their secreted protein gene expression profile, as these 

proteins have potential to be detected systematically. Through 

bioinformatic analysis, we identified genes that are specifically 

expressed by activated HSCs compared to whole liver and 

36 other human tissues (dbGaP). Further, to identify candidates 

of greater biological relevance we focused on: 1. genes 

enriched in activated versus quiescent HSCs, 2. those directly 

regulated by TGFβ, defined by upregulaton of expression by 

TGFβ and proximity to SMAD3 sites, and 3. genes within 

super-enhancers (presence of H3K27ac marks 10kb upstream 

of transcription start site). We cross-referenced candidates to 

a previously published 186-gene set that predicts outcomes of 

patients with HCC or hepatitis-C related early-stage cirrhosis. 

Additionally, we compared candidates to recently published 

and unpublished data sets corresponding to stellate signatures. 

Results: We identified a total of 218 genes enriched in activated 

HSCs. Of these 90 are upregulated by TGFβ, of which 

62 have associated SMAD3 sites, suggesting direct regulation. 

Further, 10 candidates are found within super-enhancers, 5 

of which are TGFβ-regulated and contain SMAD3 sites. All 5 

genes are implicated in liver fibrosis and may serve as putative 

biomarkers. Finally, we found that our 218 HSC-specific gene 

signature is unique and associated with negative clinical outcomes 

in patients with HCC and hepatitis C-related cirrhosis. 

Conclusion: Taken together, this work provides a functionally 

relevant list of genes and proteins that have potential as serum 

biomarkers in the non-invasive assessment of hepatic fibrogenic 

activity. 

Disclosures: 


Merck, BMS 

The following authors have nothing to disclose: Daniel L. Motola, Chan Zhou, 

Samuel York, Yujin Hoshida, Alan Mullen 

779 

Diagnostic disparity of FIB4, APRI and AAR in predicting 

liver fibrosis in HDV/HBV co-infected patients 

Varun K. Takyar 1 , David E. Kleiner 2 , Kenneth J. Wilkins 3 , Pallavi 

Surana 1 , Ohad Etzion 1 , Jay H. Hoofnagle 1 , T. Jake Liang 1 , Theo 

Heller 1 , Christopher Koh 1 ; 1 Liver Diseases Branch, NIH, Bethesda, 

MD; 2 Laboratory of Pathology, NCI, Bethesda, MD; 3 Office of the 

Director, NIDDK, Bethesda, MD 

Introduction: Information on staging of liver fibrosis is essential 

in managing chronic hepatitis B (HBV), C (HCV) and D (HDV) 

virus infections. Simple, non-invasive methods for the assessing 

fibrosis originally designed for HCV have proven usefulness 

in HBV. However, their accuracy and utility have not been 

evaluated in HDV, which involves dual virulent pathogens and 

accelerated hepatic fibrosis. Aims: To evaluate the utility of 

well established non-invasive biomarkers of fibrosis in HDV 

infection. Methods: Non-invasive biomarkers of liver disease 

(aspartate aminotransferase (AST)/alanine aminotransferase 

(ALT) ratio (AAR), AST-to-platelet ratio index (APRI), Fibrosis-4 

(FIB-4) index) were correlated with liver histology in HCV, HBV 

and HDV infected subjects. Laboratory measurements were 

obtained within 24-hours of liver biopsy. Fibrosis scores (Ishak 

and Knodell) were mapped to a F0-4 equivalent scale. After 

normalization, Mann-Whitney U test used to assess statistical 

differences of area under the receiver operator curve (AUROC) 

between the 3 biomarkers. Results: Out of 2682 unique viral 

hepatitis liver biopsies,1043 index pre-treatment liver biopsies 

with biomarkers were analyzed. The mean age was 45.4+/- 

12.4 at the time of biopsy; 688 (66%) male and 646 (61.9%) 

Caucasian. Of these, 706 (68%) were HCV, 276 (26%) were 

HBV and 61 (6%) were HDV. Histologically, HDV had the 

greatest percentage of advanced fibrosis (29%) and necroinflammation 

(35%) compared to HCV (11% and 6%, respectively) 

and HBV (16% and 7%, respectively). In HCV, FIB-4 had 

the best area under the receiver operating characteristic curve


(AUROC) distinguishing severe (F3-F4) from mild-to-moderate 

fibrosis (F0-F2); 0.89 (95% confidence interval (CI), 0.86-0.91) 

followed by APRI 0.86 (95%CI, 0.83-0.90) and AAR 0.63 

(95%CI, 0.58-0.68). Similarly, FIB-4 performed best in HBV; 

0.73 (95%CI, 0.66-0.80), followed by APRI 0.64 (95%CI, 

0.56-0.72) and AAR 0.63 (95%CI, 0.55-0.72). Biomarkers 

performed the worst in HDV; AAR 0.65 (95%CI, 0.49-0.81), 

followed by FIB-4 0.60 (95% CI, 0.42-0.78) and APRI 0.55 

(95% CI, 0.37-0.72). Conclusion: Similar to published data, 

analysis of our cohort shows that FIB-4 performs better than 

APRI and AAR in predicting clinically significant hepatic fibrosis 

in HBV and HCV infections. However, in HDV, these noninvasive 

fibrosis markers lack diagnostic accuracy probably due 

to the greater necroinflammation. Noninvasive markers should 

be used with caution in HDV infected patients. 

Disclosures: 

The following authors have nothing to disclose: Varun K. Takyar, David E. 

Kleiner, Kenneth J. Wilkins, Pallavi Surana, Ohad Etzion, Jay H. Hoofnagle, T. 

Jake Liang, Theo Heller, Christopher Koh 

780 

Collagen maturation measured by true formation of 

Type III collagen (PRO-C3) provides high diagnostic 

accuracy in alcoholic liver fibrosis: A prospective biopsy 

controlled study with 196 patients 

Aleksander Krag 1 , Bjørn S. Madsen 1 , Diana J. Leeming 2 , Maja 

Thiele 1 , Mette J. Nielsen 2 , Annette D. Fialla 1 , Ove B. Schaffalitzky 

de Muckadell 1 , Janne F. Hansen 1 , Linda S. Møller 1 , Sönke Detlefsen 

3 , Morten A. Karsdal 2 ; 1 Dept. Gastroenterology and Hepatology, 

Odense Universityhospital, Odense, Denmark; 2 Biomarkers & 

Research, Nordic Bioscience, Herlev, Denmark; 3 Dept. Pathology, 

Odense Universityhospital, Odense, Denmark 

Background In alcoholic fibrosis and early cirrhosis the majority 

of patients have normal liver function tests. Serological biomarkers 

for early detection of alcoholic fibrosis are needed to 

monitor and prevent development and complications of cirrhosis 

and to motivate persistent alcohol abstinence. During fibrogenesis, 

type III collagen is synthesized and the fibril formation 

in the extracellular matrix (ECM) is dependent on site-specific 

cleavage of the pro-peptide. Consequently, Type III collagen 

formation may represent altered collagen remodelling. PRO-C3 

is a new serum marker of type III collagen formation, targeted 

at the site where the pro-peptide of type III collagen is cleaved 

by ADAM proteases. In contrast, existing PIIINP assays detect 

total type III collagen pro-peptides. We assess the diagnostic 

accuracy of PRO-C3 for alcoholic liver fibrosis with liver biopsy 

as the gold standard. Methods: We recruited 196 at-risk 

patients (prior or current alcohol abuse, no known liver disease) 

from: (A) A high a-priori fibrosis risk group from primary 

referrals to outpatient liver clinics (126 patients); and (B) a low 

a-priori fibrosis risk group from a municipal alcohol rehabilitation 

centre (70 patients). Results: The spectrum of fibrosis was 

well covered among the 196 patients with METAVIR scores of 

F0/1/2/3/4 in 12/102/35/13/34 patients. Among the 34 

with cirrhosis, median Child-Pugh was 6 (IQR 2). Median age 

was 56 years (IQR 13). The prevalence of significant fibrosis 

(≥F2) and cirrhosis (F4) in group A were 56% and 25%, versus 

17% and 4% in group B. AUROC analyses of PRO-C3 to 

diagnose significant fibrosis (≥F2) and cirrhosis (F4) was: 1) all 

patients ≥F2 = 0.84 (95% CI 0.78-0.90), =F4 = 0.85 (0.77- 

0.92), 2) ≥F2 in high-risk group = 0.85 (0.78-0.91), =F4 in 

high risk group= 0.80 (0.71-0.90), 3) ≥F2 in low-risk group= 

0.71 (0.50-0.92), =F4 in low-risk group= 0.94 (0.84-1.00). 

A cut-off of 11.3 ng/ml ruled out cirrhosis with a sensitivity of 

97% and a NPV of 98%. Below 11.3 ng/ml only one patient 

had cirrhosis. Above 113.6 ng/ml only one patient did not 

have cirrhosis. Conclusion Collagen maturation, evidenced by 

specific type III collagen formation, is highly upregulated in 

alcoholic liver fibrosis. The diagnostic accuracy of PRO-C3 to 

diagnose significant fibrosis and cirrhosis in two independent 

populations of patients with alcohol overuse and no known 

liver disease is high. 

Diagnostic accuracy of ProC3 

Disclosures: 

Aleksander Krag - Advisory Committees or Review Panels: Norgine; Speaking 

and Teaching: Norgine 

Diana J. Leeming - Employment: Nordic Bioscience; Patent Held/Filed: Nordic 

Bioscience 

Mette J. Nielsen - Grant/Research Support: Nordic Bioscience A/S 

Morten A. Karsdal - Management Position: Nordic Bioscience; Stock Shareholder: 

Nordic Bioscience 

The following authors have nothing to disclose: Bjørn S. Madsen, Maja Thiele, 

Annette D. Fialla, Ove B. Schaffalitzky de Muckadell, Janne F. Hansen, Linda S. 

Møller, Sönke Detlefsen 

781 

Usefulness of ultrasound elastography in assessing liver 

fibrosis in patients with chronic hepatitis and the difference 

of liver stiffness between HBV-related chronic hepatitis 

and non-HBV-related chronic hepatitis. 

Takashi Nishimura 2,1 , Chikage Nakano 2,1 , Tomoko Aoki 3 , Kyohei 

Kishino 1 , Yoshihiro Shimono 1 , Kunihiro Hasegawa 1 , Ryo Takata 1 , 

Kazunori Yoh 1 , Akio Ishii 1 , Tomoyuki Takashima 1 , Yoshiyuki 

Sakai 1 , Nobuhiro Aizawa 1 , Naoto Ikeda 1 , Hiroki Nishikawa 1 , 

Yoshinori Iwata 1 , Seiichi Hirota 4 , Jiro Fujimoto 5 , Shuhei Nishiguchi 

1 , Hiroko Iijima 2,1 ; 1 Department of Internal Medicine, Division 

of Hepatobiliary and Pancreatic Disease, Hyogo College of Medicine, 

Nishinomiya, Japan; 2 Ultrasound Imaging Center, Hyogo 

College of Medicine, Nishinomiya, Japan; 3 Department of Internal 

Medicine, Yoka Hospital, Yabu-city, Japan; 4 Department of Surgical 

Pathology, Hyogo College of Medicine, Nishinomiya, Japan; 

5 Department of Surgery, Hepato-biliary-pancreas Surgery, Hyogo 

College of Medicine, Nishinomiya, Japan 

Background and aims: Liver biopsy remains the reference standard 

in assessing liver fibrosis though it is an invasive method. 

Many studies have shown that ultrasound elastography have 

good diagnostic accuracy for the assessment of liver fibrosis 

in various chronic hepatitis. We evaluated liver stiffness 

in patients with various chronic hepatitis using shear wave 

elastography, and found out that the difference of etiologies 

was associated with liver stiffness difference in patient with 

liver cirrhosis. We compared liver stiffness in patients with different 

types of chronic hepatitis using Virtual Touch Quantification 

(VTQ, Siemens, ACUSON S2000/3000®). Methods: 

1406 patients with chronic liver disease who had undergone 

VTQ measurements from October 2008 to November 2014, 

and subsequently underwent liver biopsy within 4 months after 

VTQ measurement were evaluated to find relationship between 

VTQ and other parameters including the difference of etiology. 

This study was approved by the hospital ethics committee in 

our institution. Results: Multivariate analysis for the association 

with VTQ demonstrated that F parameter, A parameter, 

etiology HBV, platelet counts, total bilirubin, serum albumin, 

γ-GTP, PT-INR and fasting plasma glucose were associated 

with VTQ(p


positive correlation with VTQ(P


784 

Predicting clinical outcomes in chronic liver disease: The 

ELF test is superior to histology and simple scores 

Katharine Irvine 1 , Leesa F. Wockner 2 , Mihir Shanker 1 , Kevin 

Fagan 1 , Leigh Horsfall 1 , Linda Fletcher 3 , Jacobus Ungerer 4 , Carel 

J. Pretorius 4 , Gregory Miller 1 , Andrew D. Clouston 1 , Guy Lampe 4 , 

Elizabeth E. Powell 1,3 ; 1 Centre for Liver Disease Research, The 

University of Queensland, Brisbane, QLD, Australia; 2 Statistics 

Unit, QIMR Berghorfer Medical Research Institute, Brisbane, QLD, 

Australia; 3 Department of Gastroenterology and Hepatology, Princess 

Alexandra Hospital, Brisbane, QLD, Australia; 4 Pathology 

Queensland, Brisbane, QLD, Australia 

Background: Early identification of subjects at-risk of progressive 

liver disease is important because these people require 

specialist care and surveillance for liver cancer and decompensation. 

However current tools for risk stratification of chronic 

liver disease (CLD) subjects are limited. Aims: We aimed to 

determine whether the serum-based Enhanced Liver Fibrosis 

(ELF) test predicted liver-related clinical outcomes, or progression 

to advanced liver disease, and to compare the performance 

of ELF to liver biopsy and non-invasive algorithms in 

predicting clinical outcomes. Methods: 300 patients with ELF 

scores assayed at the time of liver biopsy were followed up 

for liver-related clinical outcomes (ascites, encephalopathy, 

variceal bleeding, liver cancer) by review of medical records 

and clinical data. Fibrosis was staged from liver biopsy using 

a modified METAVIR score, and patients were grouped into 

non-advanced (F0-2) and advanced fibrosis (F3-4) categories. 

Primary outcomes were liver related morbidity and mortality 

and, in the F0-2 cohort, clear evidence of progression to 

advanced fibrosis. Results: During a median follow-up period 

of 6.1 years, hepatic decompensation (n=8) or HCC (n=8) 

occurred in 14 of 73 (19.2%) patients with ELF score ≥9.8 (the 

manufacturer’s cut-off for advanced fibrosis, on average 10.2 

years after recruitment), and in 2 of 227 ( 3 was 0.930 [cutoff 9.2kPa, sensitivity 

(Se) 91.5%, specificity (Sp) 85.2%), 0.789 (Se 64.7%, Sp 

85.5%), 0.684 (Se 59.6%, Sp 68.2%) and 0.714 (Se 57.7%, 

Sp 81.7%), respectively. Similarly, the AUROCs for detecting 

F4 using SSI, FIB-4, AAR, APRI was 0.958 (Cutoff 11.4 kPa, 

Se 94.9%. Sp 85.5%), 0.864 (Se 69.7%, Sp 95.7%), 0.773 

(Se 72.0%, Sp 70.4%) and 0.771 (Se 56.6%, Sp 90.3%), 

respectively. Pairwise comparison of AUROCs showed that 

SSI demonstrated significantly better diagnostic performance 

for detecting F3-4 and F4 stage fibrosis when compared to 

all other clinical scoring systems presented here. Conclusions: 

Our data indicates that SSI is a valuable diagnostic tool for 

the diagnosis of severe fibrosis (F3-4) and liver cirrhosis with 

AUROCs greater than 0.9 that demonstrated higher accuracy 

than several routinely used clinical scoring systems. 

Disclosures: 






The following authors have nothing to disclose: Masato Yoneda, Emmanuel 

Thomas, Tiffannia Grant, Seth N. Sclair


786 

Comparative 10-Years Prognosis of Fibrotest (FT) and 

Liver Stiffness Measurement (LSM) by Transient Elastography 

(TE, Fibroscan) in 9364 Chronic Liver Diseases 

(CLD) Patients (Pts) 

Mona Munteanu 1,2 , Yen Ngo 2 , Hugo Perazzo 3 , Pascal Lebray 3 , 

Helmi Mkada 4 , Olivier Deckmyn 2 , Denis Monneret 4 , Françoise 

Imbert-Bismut 4 , Dominique Bonnefont-Rousselot 4 , Vincent Thibault 

5 , Vlad Ratziu 3 , Chantal Housset 6 , Thierry Poynard 3 ; 1 University 

Pierre and Marie Curie – Univ Paris 06 UMR_S 938, Paris, 

France; 2 Biopredictive Hepatology Resarch Unit, Paris, France; 

3 UPMC Liver Center APHP, Paris, France; 4 Biochemistry Department, 

UPMC Liver Center, Paris, France; 5 Virology Department, 

Hôpital Pitié-Salpêtrière, AP-HP, Paris, France; 6 Centre de Recherche 

Saint-Antoine & Institute of Cardiometabolism and Nutrition 

(ICAN), INSERM & UPMC –Univ Paris 06 UMR_S 938, Paris, 

France 

FT and LSM were validated for fibrosis and cirrhosis staging: 

F4.1-no complications, F4.2-varices, F4.3-decompensated. 

(JHepatol2014) Aim.To assess comparatively FT and LSM 

10-years survivals, overall (OS) and without liver-related deaths 

(SLD), according to the severity of fibrosis/cirrhosis in a prospective 

CLD cohort. Methods. Cut-offs for F4.1/F4.2/F4.3 

were: for FT 0.74/0.85/0.95 and LSM(kPa)12.5/20/50. 

Death(D) and liver transplantation(LT) rates were collected 

from the national and hospital registries.Statistics used the 

time to event Cox proportional-hazard. Results. N=9364pts 

with FT were included between 2003-2010: 60%males, 

age48.9yrs; F0F1 68%,F2F3 17% and F4 15%;etiologies: 

alcohol 6%, virus B 24%, C 45%, non-alcoholic fatty 

liver 10%,other15%; mean(max) follow-up 51(121)months. 

N=2724pts had applicable LSM. The OS %(95%CI)/%D as 

per FT decreased significantly with fibrosis stage increase 

(all p


p


(NPV 99%). To rule in cirrhosis, the optimal cut off values were 

51.4 and 27.3 for TE and 2D-SWE. Above these limits, only 

four patients did not have cirrhosis on biopsy (PPV 86% for TE 

and 84% for 2D-SWE). Conclusion: TE and 2D-SWE can be 

used as screening tools for alcoholic cirrhosis, with higher cutoff 

values than for HCV cirrhosis. Elastography reliably predicts 

the absence of cirrhosis. However, the risk of having cirrhosis 

in case of a positive test varies substantially according to the 

population’s disease prevalence. The positive predictive value 

should be taken into account when interpreting liver stiffness 

results. 

Disclosures: 

Aleksander Krag - Advisory Committees or Review Panels: Norgine; Speaking 

and Teaching: Norgine 

The following authors have nothing to disclose: Maja Thiele, Bjørn S. Madsen, 

Janne F. Hansen, Sönke Detlefsen, Linda S. Møller, Annette D. Fialla 

790 

Comparison of noninvasive fibrosis scores and association 

with mortality in adults with moderate to severe 

hepatic steatosis and NAFLD 

Benjamin D. Renelus 1 , Fengxia Yan 2 , Michael C. Flood 3 ; 1 Internal 

Medicine, Morehouse School of Medicine, Atlanta, GA; 2 Clinical 

Research Center, Community Health and Preventive Medicine, 

Morehouse School of Medicine, Atlanta, GA; 3 Division of Gastroenterology, 

Morehouse School of Medicine, Atlanta, GA 

Background: Non-alcoholic fatty liver disease (NAFLD) is highly 

prevalent, causing the majority of elevated serum aminotransferases. 

NAFLD is a disease spectrum ranging from steatosis 

to steatohepatitis. Steatohepatitis, which requires liver biopsy 

for confirmation, may progress to fibrosis, cirrhosis and liver 

cancer, thus contributing to overall morbidity and mortality. 

Knowledge regarding non-invasive long-term prognostic factors 

of NAFLD is limited. Aim: The purpose of this study is to evaluate 

accuracy of non-invasive fibrosis markers and scores with 

mortality among adults with moderate to severe steatosis and 

NAFLD. Methods: We conducted a cross-sectional analysis of 

data derived from the Third National Health National Health 

and Nutrition Examination Survey 1988-1994 (NHANES III) 

and ensuing follow-up mortality data through December of 

2006. NAFLD was defined as ultrasonographic findings consistent 

with moderate to severe hepatic steatosis in the absence 

of viral hepatitis, or excessive alcohol use. Excessive alcohol 

use was defined as >/=2 drinks/day in males, and >/=1 

drinnk/day in females. Mortality was paired with noninvasive 

markers of inflammation and fibrosis. These included NAFLD 

fibrosis score (NFS), Fibrosis-4 score (FIB-4), aspartate aminotransferase 

to platelet ratio index (APRI), C-reactive protein 

(CRP) and serum vitamin D (VDP). Results: A total of 11,490 

adults between the ages of 20-74 who underwent abdominal 

ultrasound were found to have NAFLD. Of those, 2641 were 

found to have moderate to severe steatosis based on ultrasonographic 

findings. Among those with moderate to severe 

steatosis there were 549 documented deaths. The vast majority 

of secondary causes were related to cardiovascular disease 

(213), followed by non-cancer liver related deaths (8) and 

hepatocellular carcinoma (HCC) (4). The diagnostic threshold 

for NAFLD among the inflammatory and fibrosis markers were 

as follows: VDP level 3mg/L, APRI >0.98, 

NFS >0.676, and FIB-4 >3.25. The area under the receiver 

operating characteristic curve (ROC curve) for the noninvasive 

markers with all-cause mortality were NFS 0.722, FIB-4 0.733, 

CRP 0.559, APRI 0.515, VDP 0.517. P-value were significant 

for all markers except for APRI and VDP. Conclusion: FIB-4 has 

strongest association with mortality among adults with ultrasound 

based moderate to severe NAFLD. FIB-4 may have some 

prognostic role in those with advanced NAFLD however further 

studies are necessary to confirm. 

Disclosures: 

The following authors have nothing to disclose: Benjamin D. Renelus, Fengxia 

Yan, Michael C. Flood 

791 

Effect of inflammation on liver stiffness in active autoimmune 

liver disease: A simultaneous biopsy-controlled 

study using Acoustic Radiation Force Impulse (ARFI) 

elastography 

David I. Sherman 1 , Waleed Fateen 1 , Minal J Sangwaiya 2 , Paul 

Tadrous 3 , Philip J. Shorvon 2 ; 1 Gastroenterology, Central MIddlesex 

Hospital, London North West Healthcare NHS Trust, London, 

United Kingdom; 2 Radiology, Central Middlesex Hospital, London 

North West Healthcare NHS Trust, London, United Kingdom; 

3 Cellular Pathology, Northwick Park Hospital, London North West 

Healthcare NHS Trust, London, United Kingdom 

Introduction ARFI elastography (virtual touch quantification, 

VTq) is a well validated technique for non-invasive assessment 

of liver fibrosis in viral hepatitis. The interpretation of 

shear velocity readings in active autoimmune liver disease 

(AILD) may be affected by a number of factors. However, few 

studies have specifically examined the effect of inflammation 

on liver stiffness (LS) in this group. We report the results of a 

preliminary study in which LS and histology have been sampled 

simultaneously from the same region of liver tissue in a large 

cohort with active AILD. Patients and Methods Our local database 

of 101 patients with AILD (63 autoimmune hepatitis AICH 

+/- overlap, 38 cholestatic - PBC or PSC) was investigated. LS 

estimation by ARFI was performed using a standard validated 

protocol by a single operator. Biopsies were performed from 

the same region of liver using an 18G Biopince needle, 

immediately after LS measurement. Clinical, biochemical, ultrasonic 

and histopathological data were collated retrospectively. 

ARFI/histological variance (AHV) was defined as a difference 

of more than 1 Metavir or 2 Ishak stages from that predicted 

by ARFI, according to standard calibration. 1 Results Sixty one 

ARFI + liver biopsies performed at the same session were identified 

out of a total of 164 ARFI and 114 liver biopsies. Patients 

included group 1: 34 active AICH (diagnosis / flare on therapy); 

group 2: 7 AICH remission; and group 3: 17 cholestatic 

liver disease. Validation confirmed satisfactory ARFI quality: 

SD/mean > 0.3 in 4(6.9%), failure in 3(4.9%). Co-pathology 

was seen in 8(13%), mostly NAFLD. Mean ARFI shear velocities 

in groups 1, 2 and 3 were 2.41, 1.29 and 1.64 m/sec; AHV 

occurred in 44.1, 28.6 and 29.4%, respectively. AHV prevalence 

was 41.4% overall, with 100% in group 1 and 88% 

overall reflecting overestimation of fibrosis. Across all groups, 

Ishak necro-inflammatory grade was strongly correlated with 

both ARFI shear velocity (r=0.58,p


Disclosures: 

The following authors have nothing to disclose: David I. Sherman, Waleed 

Fateen, Minal J Sangwaiya, Paul Tadrous, Philip J. Shorvon 

792 

2D Shear Wave Elastography is not Superior to Transient 

Elastography in Difficult-to-Scan Patients: A Comparative 

Feasibility Study 

Benjamin Staugaard 3 , Janne F. Hansen 3 , Belinda Mössner 3 , 

Bjørn S. Madsen 1 , Aleksander Krag 1 , Peer B. Christensen 3 , 

Maja Thiele 1,2 ; 1 Department of Gastroenterology and Hepatology, 

Odense University Hospital, Odense C, Denmark; 2 OPEN, 

Odense Patient data Exploratory Network, Odense University 

Hospital, Odense, Denmark; 3 Department of Infectious Diseases, 

Odense University Hospital, Odense, Denmark 

Introduction: 2D shear wave elastography (2D-SWE) is a promising 

new elastography technique. Some studies suggest that 

2D-SWE reduce the number of invalid measurements otherwise 

seen with transient elastography (TE), but a direct comparison 

between the two techniques in difficult-to-scan patients 

has not been performed. Methods: Single-center comparative 

study to evaluate the feasibility of TE (Echosens FibroScan) and 

2D-SWE (Supersonic Aixplorer). We included patients with 

hepatitic C and alcoholic liver disease with a invalid TE at 

their latest appointment. At inclusion, an experienced operator 

re-examined patients with same-day TE and 2D-SWE. TE was 

evaluated using standard quality criteria, while 2D-SWE quality 

criteria mimicked TE criteria: SD/mean should be ≥ 30%. 

Additionally, we judged 2D-SWE image quality subjectively: 

A uniform colour-coded region of interest should remain stable 

for 3 seconds and be at least 15 mm in diameter. Results: 

From 10,247 TE examinations performed between 2007-14, 

we identified 119 eligible patients and included 52 in the 

study. The median age was 57 years and 60% were male. 

The majority of patients were overweight (median BMI 31 kg/ 

cm 2 ; range 19-55; BMI ≥ 30 in 54% of patients). None of the 

patients had ascites. The initial screening examination was a 

failure in 13 of patients, while 39 patients had a unreliable 

TE. At re-examination, TE and 2D-SWE were both feasible in 

33 patients (63%). Seventeen patients had a valid TE, but an 

invalid 2D-SWE (33%). In two patients, both TE and 2D-SWE 

failed (4%). Both these patients were investigated with the XL 

probe. The XL probe was used in a total of 31 patients. The 

applicability of TE was superior to 2D-SWE, as TE was valid 

in 96% of re-examined patients while 2D-SWE was valid in 

63% (P





The following authors have nothing to disclose: Brian J. Hogan, James O’Beirne, 

David W. Patch, Dominic Yu, Ioanna Parisi, Andrew R. Hall, Ankur Srivastava, 

Paul M. Trembling, Sudeep Tanwar 

794 

Association of skin capsular distance with accuracy 

of liver stiffness measurements of the FibroScan XL 

probe—comparison with virtual touch quantification 

and the FibroScan M probe 

Erina Kumagai 1,3 , Keiko Korenaga 2 , Masaaki Korenaga 1,2 , 

Misuzu Ueyama 1,3 , Yoshihiko Aoki 2 , Yoko Yamagiwa 1 , Masatoshi 

Imamura 2 , Kazumoto Murata 1,2 , Tatsuya Kanto 1,2 , Naohiko 

Masaki 1,2 , Sumio Watanabe 3 , Masashi Mizokami 1,2 ; 1 The 

Research Center for Hepatitis and Immunology, National Center 

for Global Health and Medicine at Kohnodai, Ichikawa, Japan; 

2 Department of Gastroenterology and Hepatology, Kohnodai Hospital, 

National Center for Global Health and Medicine, Ichikawa, 

Japan; 3 Department of Gastroenterology, Juntendo University 

School of Medicine, Bunkyo-ku, Japan 

Background and Aim: The FibroScan XL probe was specifically 

designed for obese patients. However, there is no data 

regarding its use in Japanese individuals, who tend to be less 

obese than Caucasians. This study evaluates the diagnostic 

performance of the XL probe, the FibroScan M probe, and the 

virtual touch quantification (VTQ) method in Japanese patients 

with chronic liver disease. Methods: A total of 664 consecutive 

patients with chronic liver disease who underwent ultrasonography 

were prospectively enrolled from June through October 

2014. The cause of liver disease was hepatitis C virus (HCV, 

n = 179), hepatitis B virus (HBV, n = 79), fatty liver (n = 264), 

or non-HBV non-HCV hepatitis (n = 142). Liver stiffness was 

measured by VTQ, the M probe, and the XL probe on the same 

day. Liver stiffness measurement (LSM) failure was defined as 

zero valid shots; unreliable examination was defined as


796 

Glyco-isomer of Serum Mac-2-Binding Protein is The 

Most Useful Biomarker for Liver Fibrosis in Patients with 

Chronic Hepatitis C 

Mina Hamano, Masafumi Naito, Toshifumi Ito; Japan Community 

Healthcare Organization Osaka Hospital, Osaka, Japan 

Background and Aim The degree of liver fibrosis is a major 

issue for making decision of therapeutic strategies on chronic 

liver diseases. Recently glyco-isomer of serum Mac-2-binding 

protein (M2BPGi) could be applied as a novel and useful biomarker 

to assess liver fibrosis. However little is known about 

its usefulness for diagnosis of chronic hepatitis C patients. In 

this study, we were trying to reveal the significance of serum 

M2BPGi for assessment of liver fibrosis. Method One hundred 

fifty patients with HCV infection were enrolled, one hundred 

nine patients were scheduled for the anti-viral therapy or eleven 

patients had received hepatectomy for hepatocellular carcinoma. 

Liver histology was assessed in all of the patients. We 

evaluated the relation between liver fibrotic degrees and the 

characteristics of the patients by Kruskal-Wallis test. And the 

correlation between the serum M2BPGi and examined liver 

function tests, i.e. platelet, T-Bil, AST, ALT, GGT, albumin, PT, 

AFP, γ-globulin and hyaluronic acid (HA). Additionally we evaluated 

the ability of M2BPGi by ROC analysis compared with 

HA and Fib4 index. The multiple logistic regression analysis 

was performed to evaluate the contribution of fibrosis (fibrosis 

stage ≥ F2). Results The patients were classified into four 

groups according to liver fibrosis stage, F0 (n=34), F1 (n=53), 

F2 (n=39), and F3-4 (n=24). The serum M2BPGi were significantly 

increased in relation to liver fibrosis, (F0/F1/F2/ 

F3-4 median=0.85/1.30/2.13/2.75C.O.I., p


798 

Machine-learned Image Analysis Models for Classifying 

Liver Fibrosis Stage from Magnetic Resonance Images 

Timothy G. St. Pierre 1 , Michael J. House 1 , Ajmal Mian 2 , Sander 

Bangma 3 , Gary Burgess 6 , Richard A. Standish 4 , Stephen Casey 5 , 

Emma Hornsey 7 , Peter W. Angus 5 ; 1 School of Physics M013, University 

of Western Australia, Crawley, WA, Australia; 2 School of 

Computer Science and Software Engineering, The University of 

Western Australia, Crawley, WA, Australia; 3 Resonance Health 

Ltd, Claremont, WA, Australia; 4 School of Medicine, Deakin University, 

Waurn Ponds, VIC, Australia; 5 Liver Transplant Unit, Austin 

Hospital, Heidelberg, VIC, Australia; 6 Pfizer Inc, New York, NY; 

7 Department of Radiology, Austin Hospital, Heidelberg, VIC, Australia 

The aim of the study was to assess the potential of machinelearned 

image analysis models to classify liver fibrosis stage 

in patients with hepatitis C virus (HCV). Methods: Patients with 

liver fibrosis due to chronic HCV infection were recruited from 

the Victorian Liver Transplant Unit. All subjects had undergone 

a liver transplant for HCV cirrhosis at least 12 months prior 

to the screening visit, and each subject had also had a liver 

biopsy in the previous 6 months, with a fibrosis stage score 

of METAVIR F1 - F3. Twenty eight subjects were recruited and 

scanned with MRI at two visits (visit 1 and visit 2). Visit 2 was 

planned to occur between 35 and 45 days after visit 1. The 

distribution of Ishak stage scores on the re-examined pre-study 

biopsies was 0 (n=2); 1 (n=5); 2 (n=6); 3 (n=8); 4 (n=2); 5 

(n=3); 6 (n=2). A 1.5 T Siemens Avanto MR scanner was used 

to acquire axial images of the abdomen. A series of breathhold 

spoiled gradient echo images was acquired before and 

after a bolus injection of 10 mL or 0.1 mmol/kg (whichever 

was lower) of gadoterate (Dotarem®) contrast agent including 

acquisitions at 3 minutes and 10 minutes after injection. 

Machine learning algorithms were trained on the images from 

visit 1 to classify subjects into Ishak stage 2 or below and 3 or 

above. These algorithms extract rotation invariant features by 

quantizing gradients from local neighbourhoods of key locations 

of the images at different scales. The most relevant feature 

statistics were then automatically selected to train Support Vector 

Machine (SVM) classifiers. The potential of the algorithms to 

correctly classify patients was tested by training the algorithms 

on 27 of the 28 image sets from visit 1 and testing the resulting 

model on the 28th image set. This “leave-one-out” strategy was 

used to classify all 28 image sets from visit 1. A model trained 

on all 28 subjects from visit 1 was then used to classify all of the 

image sets obtained at visit 2 in order to assess repeatability of 

the test. Results: Using the leave-one-out models trained on 27 

of the 28 visit 1 images sets, 27, 28, and 28 out of 28 cases 

were correctly classified in visit 1 data for pre-, 3-mins post, 

and 10 mins post-contrast images. The model trained from all 

28 visit 1 datasets correctly classified 23 out of 27, and 26 out 

of 28, and 25 out of 28 cases for the pre-, 3-mins post, and 

10 mins post-contrast images from visit 2 (corresponding to 

sensitivities and specificities of 80% & 92%, 93% & 92%, and 

93% & 85% respectively. Conclusions: Machine-learned image 

analysis models have the potential to classify liver fibrosis stage 

from high contrast MR images of the liver in patients with HCV. 

Disclosures: 

Timothy G. St. Pierre - Consulting: Resonance Health Ltd; Patent Held/Filed: 

Resonance Health Ltd; Stock Shareholder: Resonance Health Ltd 

Michael J. House - Consulting: Resonance Health; Patent Held/Filed: Resonance 

Health 

Sander Bangma - Employment: Resonance Health Analysis Services Pty Ltd; Stock 

Shareholder: Resonance Health Ltd 

Gary Burgess - Employment: Pfizer, Conatus 



The following authors have nothing to disclose: Ajmal Mian, Richard A. Standish, 

Stephen Casey, Emma Hornsey 

799 

Real-time shear wave elastography is effective for 

evaluating liver fibrosis in patients with chronic liver 

diseases 

Kaori Muromachi 1 , Tsutomu Tamai 1 , Kohei Oda 1 , Sho Ijuin 1 , 

Hiroka Onishi 1 , Haruka Sakae 1 , Akihiko Oshige 1 , Kotaro Kumagai 

1 , Seiichi Mawatari 1 , Akihiro Moriuchi 2 , Hirofumi Uto 1,3 , Akio 

Ido 1 ; 1 Digestive and Lifestyle Diseases, Department of Human and 

Environmental Sciences, Kagoshima University Graduate School of 

Medical and Dental Sciences, Kagoshima, Japan; 2 Department of 

HGF Tissue Repair and Regenerative Medicine, Kagoshima University 

Graduate School of Medical and Dental Sciences, Kagoshima, 

Japan; 3 Center of Digestive and Liver Disease, Miyazaki Medical 

Center, Miyazaki, Japan 

Objective: Since liver fibrosis is the most important risk factor 

for liver cancer and mortality, accurate assessment of its severity 

is crucial. Liver biopsy is the gold standard for evaluating 

liver fibrosis, but it is an invasive method. Non-invasive assessment 

is desirable. Real-time shear wave elastography (SWE) 

installed on Logiq E9 ultrasound imaging system developed 

by GE Healthcare (USA) is a new method for evaluating liver 

fibrosis, but its usefulness has not been fully elucidated. We 

examined the utility of SWE as a new method for evaluating 

liver fibrosis. Methods: Forty-two patients with biopsy-proven 

liver disease and eighteen patients diagnosed clinically with 

liver cirrhosis were enrolled in this study. We examined the 

diagnostic performance of SWE for identifying liver fibrosis, 

as well as liver fibrosis markers and scores. Results: Twenty 

patients were male, and the median age was 63.0 years. 

The etiology of liver disease was hepatitis B virus (HBV) in 

3 patients, hepatitis C virus (HCV) in 20, and non-B non-C 

(NBNC) in 37. The median platelet count was 15.0×10 4 / 

mL, serum ALT was 46 IU/L, hyaluronic acid was 145.9 ng/ 

mL, type IV collagen 7S was 7.9 ng/mL, and procollagen III 

peptide was 0.9 U/mL. There were 30, 8, 3, and 19 patients 

with liver fibrosis scores of F0–1, F2, F3, and F4, respectively. 

Median SWE (m/s) in patients with F0–1/F2/F3/F4 fibrosis 

was 1.38/1.64/1.77/1.73. SWE increased with increasing 

severity of liver fibrosis and was significantly correlated with 

liver fibrosis markers (platelet count, hyaluronic acid, type IV 

collagen 7S, procollagen III peptide) and scores (FIB4 index, 

APRI). SWE was also significantly associated with liver histopathology 

findings (r=0.661, P


800 

Role of Acoustic Radiation Force Impulse (ARFI) elastography 

in non-invasive assessment of fibrosis in active 

autoimmune liver disease: A simultaneous biopsy-controlled 

study 

David I. Sherman 1 , Waleed Fateen 1 , Minal J Sangwaiya 2 , Paul 

Tadrous 3 , Philip J. Shorvon 2 ; 1 Gastroenterology, Central MIddlesex 

Hospital, London North West Healthcare NHS Trust, London, 

United Kingdom; 2 Radiology, Central Middlesex Hospital, London 

North West Healthcare NHS Trust, London, United Kingdom; 

3 Cellular Pathology, Northwick Park Hospital, London North West 

Healthcare NHS Trust, London, United Kingdom 

Introduction The predictive accuracy of ARFI elastography (virtual 

touch quantification, VTq) for the non-invasive assessment 

of liver fibrosis is well validated in viral hepatitis. Whilst there 

is evidence that inflammation can increase liver stiffness, the 

role of ARFI in interpreting active autoimmune liver disease is 

unclear. We report the results of a preliminary biopsy-controlled 

study in a large autoimmune cohort, in which liver stiffness 

(LS) and histology were sampled simultaneously from the same 

region of liver tissue. Patients and Methods Our local database 

of 101 patients with autoimmune liver disease (63 AICH +/- 

overlap, 38 PBC or PSC) was interrogated. LS estimation by 

ARFI was performed using a standard validated protocol by a 

single operator. Biopsies were performed from the same region 

of liver using an 18G Biopince needle, immediately after LS 

measurement. Clinical, biochemical, ultrasonic and histopathological 

data were collated retrospectively. Predictive accuracy 

variables were determined for fibrosis stage using both standard 

and local calibrations 1 . Results Sixty one ARFI + liver 

biopsies performed at the same session were identified out of a 

total of 164 ARFI examinations and 114 liver biopsies. Patients 

included group 1: 34 active AICH (diagnosis pre-therapy or 

flare on therapy); group 2: 7 AICH biochemical remission; and 

group 3: 17 cholestatic liver disease. Validation confirmed 

satisfactory ARFI quality: SD/mean > 0.3 in 4(6.9%), failure 

in 3(4.9%). Co-pathology was seen in 8(13%), mostly NAFLD. 

AUROC analysis showed overall predictive performance for 

all groups/group 1/group 3 were 54.8/41.7/56.3, respectively; 

for exclusion of ≥F2 69.5/44.9/81.8; for detection of 

F3/4 fibrosis 70.1/55.9/79.8. Using the chosen cut off point, 

sensitivity and NPV for F3/4 detection were 100% for both all 

groups and group 3. No difference was seen between performance 

of standard and local calibrations. Conclusion These 

simultaneous paired data demonstrate a reduced predictive 

performance for fibrosis stage using ARFI in active autoimmune 

liver disease, probably due to the inflammatory process. However, 

reasonable performance was demonstrated for both ≥F2 

exclusion and confirmation of F3/4 in PBC/PSC. The finding 

that ARFI can reliably exclude advanced fibrosis, particularly 

in cholestatic liver disease, is of potential practical importance. 

Further biopsy-controlled studies are needed to confirm the role 

of ARFI in this patient group. Reference D. Sherman et al. J 

Hepatol 2014;60(1):Suppl. p S413. 

Disclosures: 

The following authors have nothing to disclose: David I. Sherman, Waleed 

Fateen, Minal J Sangwaiya, Paul Tadrous, Philip J. Shorvon 

801 

A clustering based fully automated method for collagen 

proportional area extraction in liver biopsy images 

Nikos Giannakeas 2 , Zoi Tsianou 2 , Markos Tsipouras 2 , Alexandros 

T. Tzallas 3 , Pinelopi Manousou 1 , Epameinondas Tsianos 2 ; 

1 Institute for Liver and Digestive Health, London, Royal Free Hospital 

and UCL, London, United Kingdom; 2 1st Division of Internal 

Medicine and Division of Gastroenterology, Faculty of Medicine, 

School of Health Sciences, University of Ioannina, Ioannina, 

Greece; 3 School of Applied Technology, Department of Computer 

Engineering, Technological Educational Institute of Epirus, Arta, 

Greece 

Background and Aim: Collagen Proportional Area (CPA) 

extraction using digital image analysis (DIA) in liver biopsies 

provides an effective way to estimate liver disease staging. 

CPA represents accurately fibrosis expansion in liver tissue 

compared to semiquantitative staging scores. However, CPA 

has not reached everyday clinical practice. The lack of standardized 

and robust methods for computerized DIA for CPA 

assessment is a major limitation. We aim to create a fully 

automated methodology for CPA computation in liver biopsy 

images. Method: The proposed methodology is based in 

three stages. 1)The tissue detection stage: a k–means clustering 

approach is used to separate the tissue from the background. 

2)The tissue characterization stage: machine learning 

techniques are employed to characterize liver tissue, muscle, 

vessels, blood clots, structural collagen, dye stain and artifacts 

(scratches or contaminations of the substrate). Several shape 

and color features are extracted from each tissue area. This set 

of features is used for training and testing a classification algorithm, 

for automated area classification. 3) The CPA calculation 

stage: this stage also employs the k-means clustering, to separate 

fibrosis areas from normal liver tissue. CPA is computed as 

the number of fibrosis pixels divided by the number of pixels 

of the whole liver tissue. Results: The proposed methodology 

was evaluated using a dataset of 93 images of liver biopsies, 

obtained from 93 hepatitis C patients. The dataset was kindly 

provided by the Royal Free Hospital, London and our results 

were compared with the standardized method. Liver biopsies 

were formalin fixed, paraffin embedded and stained using 

Picrosirius red. High accuracy results were obtained for all 

stages of the proposed methodology (1 st stage: 98% accuracy 

in tissue detection, 2 nd stage: 85% accuracy in tissue area 

characterization, and 3 rd stage: less than 2% mean error). Conclusions: 

According to the results, CPA is extracted accurately 

in most of the cases (errors


802 

Feasibility of three ultrasound shear wave elastographic 

methods in overweight and obese patients 

Ioan Sporea; University of Medicine and Pharmacy Timisoara, 

Timisoara, Romania 

In the last decade, different types of ultrasound based elastographic 

methods that noninvasively quantify liver fibrosis 

have been developed. Even though, Transient Elastography 

is a validated method for liver fibrosis assessment in chronic 

B and C hepatitis, reliable elasticity measurements are difficult 

to obtain in obese (BMI>30kg/m 2 ) patients. The aim of this 

study was to compare the feasibility of three elastrographic 

methods used for liver fibrosis evaluation (Transient Elastography-TE, 

point Shear Wave Elastography (pSWE) using ARFI 

technique - VTQ and SuperSonic Shear Imaging-2D-SWE) in 

overweight and obese patients. Material and methods: The 

study included 183 consecutive subjects with or without chronic 

hepatopathies, in which liver stiffness (LS) was evaluated in 

the same session by means of 3 elastographic methods: TE 

(Fibroscan, Echosens), VTQ (Siemens Acuson S200 TM ) and 

2D-SWE (Aixplorer, SuperSonic Imagine S. A). Reliable LS 

measurements were defined as follows: for TE and VTQ – the 

median value of 10 LS measurements with a success rate≥60% 

and an interquartile range 25kg/m 2 ), 32.2 % with 

obesity grade 1 (BMI=30-34.9 kg/m 2 ), 4.3% with obesity 

grade 2 (BMI=35-39.9 kg/m 2 ) and 1.1% with obesity grade 

3 (BMI>40 kg/m 2 ). From the 114 overweight patients, reliable 

LS measurements (LSM) were obtained in 83.3% (95%) 

by means of TE, 85.1% (97) by means of VTQ and in 88.6% 

(101) by 2D-SWE. TE, 2D-SWE and VTQ had similar rates 

of reliable LSM in overweight patients: TE (83.3%) vs. VTQ 

(85.1%) p=0.84; TE (83.3%) vs. 2D-SWE (88.6%) p=0.33; 

2D-SWE (88.6%) vs. VTQ (85.1% ) p=0.55. The percentage of 

reliable LSM in obese patients was similar: VTQ (79.7%) vs TE 

(82.6%) (p=0.82), 2D SWE (85.5%) vs TE (82.6%) p=0.81, 

VTQ (79.7%) vs 2D SWE (85.5%) p=0.5 Conclusion: Feasibility 

of TE, VTQ and 2D-SWE in overweight and obese patients 

was similar. 

Disclosures: 

The following authors have nothing to disclose: Ioan Sporea 

803 

Fast macromolecular proton fraction (MPF) mapping of 

the human liver in vivo for quantitative assessment of 

hepatic fibrosis 

Vasily L. Yarnykh 1,2 , Erica Tartaglione 3 , George N. Ioannou 3,4 ; 

1 Radiology, University of Washington, Seattle, WA; 2 Research 

Institute of Biology and Biophysics, Tomsk State University, Tomsk, 

Russian Federation; 3 Research and Development, Veterans Affairs 

Puget Sound Health Care System, Seattle, WA; 4 Medicine, Division 

of Gastroenterology, University of Washington, Seattle, WA 

Background: MPF is a quantitative MRI parameter determining 

the magnetization transfer (MT) effect and defined as a relative 

amount of immobile macromolecular protons involved into 

magnetization exchange with mobile water protons. MPF has 

a potential for quantitative assessment of fibrous tissue due to 

intrinsically high MPF in collagen. A recent fast single-point 

MPF mapping method (Magn Reson Med 2012;68:166) 

enables the capability of liver MPF assessment. The goal of this 

study was to prospectively investigate a relationship between 

MPF in the liver parenchyma and fibrosis stage. Methods: 

16 patients with chronic HCV infection underwent MRI at 3T 

using a three-dimensional MPF mapping protocol comprising 

four breath-hold scans obtained with 2x3x6 mm 3 voxel size 

and 10 sections:1) dynamic acquisition of MT-weighted (TR/ 

TE=18.5/2.3 ms, flip angle (FA)=80, saturation frequency 2 

kHz and FA=2520) and reference (same parameters, no saturation) 

images; 2) dynamic acquisition of three images for variable 

FA T1 mapping (same TR/TE, FA=3,10,20°); 3) dual-TE 

B0 map; and 4) dual-TR B1 map. MPF maps were reconstructed 

using the single-point algorithm. 14 patients had liver 

biopsy within one year prior to study entry, and two patients 

had clinically diagnosed cirrhosis. Fibrosis was histologically 

staged according to METAVIR. Score F4 was assigned to clinical 

cirrhosis. The mode of liver MPF histograms was compared 

between groups with (F2-F4, n=6) and without (F0-F1, n=10) 

significant fibrosis by Mann-Whitney test. Association between 

MPF and fibrosis score tested using Spearman correlation coefficient 

(ρ). Results: MPF was increased in the significant fibrosis 

group compared to the no or mild fibrosis group (6.49±0.36% 

vs. 5.94±0.26%, P


7 days. Oxysterols and bile acids in the liver and brain were 

extracted, and analyzed by HPLC or LC/MS. Hepatic mRNA 

were determined by RT-PCR. Primary hepatocytes were also 

isolated from the same strains of mice. Cultures were incubated 

with [ 14 C]-cholesterol and formed [ 14 C]-oxysterols/bile acids 

were analyzed. Results: In wild type mice, hepatic CYP7B1 

mRNA expression was suppressed (>80%Ñ) after StARD1 overexpression. 

SHP mRNA expression was markedly increased 

(300%É), while CYP7A1(40%Ñ) and CYP8B1(80%Ñ) mRNAs 

were decreased. Interestingly, gallbladder bile acid concentration 

in these mice was 30% higher as compared to control 

mice. Primary hepatocyte culture experiments showed that the 

rate of bile acid synthesis increased 3-fold after StARD1 overexpression. 

These observations implied that an additional bile 

acid synthetic pathway is present which compensates for the 

CYP7A1 and the CYP27A1/CYP7B1 pathways. StARD1 overexpressed 

liver accumulated 27-Hydroxycholesterol(HC) and 

25HC. Also found was unexpected high level of 24HC(Ratio of 

24, 25, 27HC, 6:1:6). Level of 24HC in the brain was identical 

to control. When primary hepatocytes were incubated with 

[ 14 C]-cholesterol following StARD1 overexpression, wild type 

hepatocytes formed all three [ 14 C]-oxysterols, 24HC, 25HC 

and 27HC(ratio, 3:1:10). However, hepatocytes isolated from 

CYP27A1 -/- mice formed only [ 14 C]-25HC. These observations 

show that 24HC originates from liver mitochondrial CYP27A1. 

Furthermore, when StARD1 was overexpressed in CYP7B1 -/- 

mouse liver in vivo and in vitro, 27HC and 25HC accumulated 

but, not 24HC; suggesting 24HC is responsible for an alternative 

metabolic pathway to bile acids. Conclusion: The results 

indicate that hepatic CYP27A1 catalyzes not only 25- and 

27-hydroxylations, but also 24-hydroxylation. Since it has been 

reported that 24HC is metabolized by CYP7B1, CYP39A and 

CYP7A1, this oxysterol may be an important intermediate in a 

second alternative pathway to bile acids. 

Disclosures: 

William M. Pandak - Employment: Virginia Commonwealth University, Veterans 

Affairs Medical Center 

The following authors have nothing to disclose: Genta Kakiyama, Dalila M. 

Marques, Kuniko Mitamura, Hajime Takei, Hiroshi Nittono, Daniel Rodriguez-Agudo, 

Gregorio Gil, Huiping Zhou, Phillip B. Hylemon 

805 

Bile acid induced cholestatic liver injury involves TLR9 

initiated inflammatory signals in mouse hepatocytes 

Shi-Ying Cai, Xinshou Ouyang, Albert Mennone, Carol J. Soroka, 

Wajahat Z. Mehal, James L. Boyer; Yale Univ, New Haven, CT 

Background: The inflammatory response plays an important 

role in cholestatic liver injury where bile acid (BA) induction 

of proinflammatory cytokines in hepatocytes may initiate this 

event. However, the signaling pathways involving BA stimulation 

of cytokine production remain elusive, although BA injures 

mitochondria in hepatocytes. Toll-like receptors (TLR) respond 

to both endogenous sterile insults and pathogen recognitions 

and play a critical role in tissue damage initiated inflammatory 

responses. TLR9 has been characterized as an intracellular 

DNA receptor that, upon activation, stimulates cytokine production 

as part of the innate immune response. Aim: To determine 

Tlr9’s role in BA induced liver injury. Methods: Hepatocytes 

isolated from wild-type (WT) and Tlr9 knockout (KO) mice were 

treated with BAs. Mitochondrial damage was assessed using a 

JC-1 assay and Western blot. Q-PCR was used to detect inflammatory 

chemokine gene expression. WT and Tlr9 KO mice 

were subjected to bile duct ligation (BDL) or sham operation 

for 7 days. Plasma biochemistry, liver gene expression, and 

liver histology were examined. Results: At pathophysiological 

concentration (≥25 mM), taurocholic acid (TCA) caused mitochondrial 

membrane potential changes in mouse hepatocytes. 

Mitochondrial specific proteins cytochrome C, AIF and ER 

specific protein Grp78 were detected in the cytosolic fraction 

by Western blot analysis, indicating mitochondria damage 

and ER stress, whereas no ROS were detected. Cxcl2 induction 

was significantly less in Tlr9 KO hepatocytes than in WT 

hepatocytes (KO: 3.7-fold v/s WT: 14-fold) after treatment with 

100mM TCA for 24 hr. The synthetic Tlr9 ligand CpG ODNs 

and BAs demonstrated synergistic effects in stimulating Cxcl2 

expression in mouse hepatocytes, consistent with Tlr9’s involvement 

in BA induction of Cxcl2 expression. After BDL, plasma 

ALT, ALP and BA levels and liver BA levels were also significantly 

lower in Tlr9 KO mice compared to WT BDL mice, confirming 

a role for Tlr9 in mediating cholestatic liver injury. Bile 

duct proliferation, assessed by liver H&E histology and CK19 

labeling, and hepatic levels of Ccl2 and Cxcl2 mRNA were 

also significantly less in Tlr9 KO than in WT mice after BDL. 

Conclusion: BA injures mitochondria resulting in DNA release 

and triggers an innate immune response by activating Tlr9. 

Preventing mitochondrial damage or blocking Tlr9 activation 

may be new strategies for treating cholestasis. 

Disclosures: 


abbvie 

The following authors have nothing to disclose: Shi-Ying Cai, Xinshou Ouyang, 

Albert Mennone, Carol J. Soroka, Wajahat Z. Mehal 

806 

Proteomics analysis of rat canalicular membrane 

reveals the expression of several P4-ATPases in liver 

Pururawa M. Chaubey, Bruno Stieger; Department of Clinical 

Pharmacology and Toxicology, University Hospital Zurich, Zurich, 

Switzerland 

Background: Transport processes in the canalicular membrane 

are key elements in bile formation and are the driving force 

for the enterohepatic circulation of bile salts. The canalicular 

membrane is constantly exposed to a hostile environment due 

to the detergent action of the high conentration of canalicular 

bile salts. Aims: To obtain more insights into the molecular entities 

rendering the canalicular membrane resistant to the detergent 

action of bile salts, the proteome of highly purified rat 

canalicular membrane vesicles was determined. Methods: Isolated 

canalicular membrane vesicles (JCB 98:991,1983) from 

Sprague Dawley rats were stripped from adherent proteins 

(JCB 93:97,1982), deglycosylated, protease digested and subjected 

to shot gun proteomic analysis. Expression of P4-ATPases 

was confirmed by RT-PCR and Western blotting. Localization of 

P4-ATPases was carried out with immunofluorescence methodology 

on liver sections. Results: We identified 1745 unique 

proteins in canalicular vesicles. A comparative analysis with a 

renal brush border membrane proteome and liver proteomes 

revealed a large set of membrane and membrane associated 

proteins (1287) specifically identified in our study, possibly 

because we enriched for membrane proteins. Assignment to 

functional categories of the newly identified proteins revealed 

an over-representation of transporter specific categories. We 

identified several transporters out of which 19 members are 

from SLC families and 11 members are from ABC families. 

Additionally, we also identified P4-ATPases (ATP8A1, ATP8B1, 

ATP9A, ATP11A, & ATP11C) and a P5-ATPase (ATP13A1). 

Four of 5 identified P4-ATPases are not known to be expressed 

in liver. Furthermore, expression analysis of P4-ATPases using 

RT-PCR and Western blotting could verify the expression of 

identified P4-ATPase with a rather high expression of ATP11C. 

Finally, ongoing immunofluorescence analysis demonstrates


localization of some of the identified P4-ATPase at the canalicular 

membrane. Conclusions: Our data suggest that there 

is not a large common proteome between the brush border 

membrane of kidney proximal tubule cells and the canalicular 

membrane. Furthermore, the identification of several P4-AT- 

Pases in rat liver together with the previous findings on mice 

with disrupted Atp11c displaying hyperbilirubinemia (PNAS 

108:7890,2011) suggest that important proteins involved in 

hepatocellular lipid homeostasis and canalicular lipid secretion 

may await characterization. Acknowledgement: The expert 

support by Dr. B Roschitzki from the Functional Genomics Center 

in Zurich is greatly appreciated. 

Disclosures: 

The following authors have nothing to disclose: Pururawa M. Chaubey, Bruno 

Stieger 

807 

TGR5 agonists improve pancreatic beta cell mass and 

function by reprogramming alpha cells to produce GLP- 

1: a novel mechanism of action of bile acids in diabetes 

Divya P. Kumar 1 , Amon Asgharpour 2 , Faridoddin Mirshahi 2 , So 

Hyun Park 3 , Sichen Liu 3 , Yumi Imai 3 , Jerry L. Nadler 3 , Karnam S. 

Murthy 1 , Arun J. Sanyal 2 ; 1 Physiology and Biophysics, Virginia 

Commonwealth University, Richmond, VA; 2 Internal Medicine, Virginia 

Commonwealth University, Richmond, VA; 3 Internal Medicine, 

Eastern Virginia Medical School, Norfolk, VA 

BACKGROUND: Pancreatic α cells secrete glucagon (GLU) 

under euglycemic conditions and GLP-1 under hyperglycemic 

conditions. GLU and GLP-1 are derived from alternate splicing 

of a common precursor by prohormone convertase 2 (PC2) and 

PC1 respectively. Bile acids (BA) via their cognate receptors, 

TGR5, stimulate GLP-1 release in intestinal L cells. Recently, 

TGR5 have been identified on pancreatic α cells. The potential 

effects of BA on pancreatic α cells and their physiological 

relevance are unknown. HYPOTHESIS: BA, acting via TGR5 

on pancreatic α cells, enhances hyperglycemia-induced PC1 

expression thereby releasing GLP-1 which, in turn, acts as a 

paracrine to increase pancreatic β cell mass and function. 

AIMS: To examine (1) the in vivo effect (intraperitoneal) of 

TGR5 specific agonist, INT-777 on glucose homeostasis, PC1 

expression, GLP-1 release, β cell proliferation in control and 

db/db mice (2) the effect of GLP-1 receptor antagonist on 

TGR5-mediated insulin secretion in human islets, and (3) the in 

vitro effect of INT-777 on β cell proliferation. METHODS: The 

in vivo effect of INT-777 on body weight, glucose homeostasis 

(glucose and insulin tolerance test), PC1 expression (immunofluorescence 

and RT-PCR), GLP-1 release (ELISA) and β cell 

mass and proliferation (morphometric analysis of pancreatic 

sections) was examined in control and db/db mice. The in vitro 

effect of INT-777 on β cell proliferation was measured by MTT 

assay in MIN6 cells. Insulin secretion in response to INT-777 

from human islets was measured in the presence or absence of 

GLP-1 receptor antagonist, exendin (9-39) by ELISA. RESULTS: 

Treatment with INT-777 for 7 weeks attenuated the increase 

in body weight, and improved glucose tolerance and insulin 

sensitivity in db/db mice compared to control. INT-777 augmented 

PC1 expression in α cells and stimulated GLP-1 release 

from islets of db/db mice compared to control. INT-777 also 

increased pancreatic β cell proliferation and insulin synthesis 

in vivo, but had no effect on β cell proliferation in vitro. Insulin 

secretion by INT-777 in human islets cultured under high 

(25mM) glucose was inhibited by exendin (9-39). These results 

suggest that in diabetes, GLP-1 released from α cells augments 

β cell proliferation and function. CONCLUSION: In diabetes, 

BA reprogram α cells to switch from GLU synthesis to GLP-1 

synthesis by increasing PC1 expression with associated trophic 

effects on adjacent β cells and improvement in insulin sensitivity 

and hyperglycemia. Thus, these results identify a novel mechanism 

of action of TGR5 agonist INT-777 in glucose homeostasis 

and its potential use in the treatment of insulin resistance and 

type 2 diabetes. 

Disclosures: 






The following authors have nothing to disclose: Divya P. Kumar, Amon Asgharpour, 

Faridoddin Mirshahi, So Hyun Park, Sichen Liu, Yumi Imai, Jerry L. Nadler, 

Karnam S. Murthy 

808 

Higher order metabolites of heme increase under conditions 

of hyperbilirubinemia, trigger cholestasis and 

impair hepatocellular integrity 

Raphael A. Seidel 1,5 , Franziska Schleser 1 , Christoph Sponholz 1 , 

Frans Cuperus 2,3 , Sandor Nietzsche 4 , Georg Pohnert 5 , Michael 

Bauer 1 ; 1 Department of Anaesthesiology and Intensive Care 

Medicine / Center for Sepsis Control and Care, Jena University 

Hospital, Jena, Germany; 2 Hans Popper Laboratory of Molecular 

Hepatology, Division of Gastroenterology and Hepatology, 

Department of Internal Medicine III, Medical University of Vienna, 

Vienna, Austria; 3 Pediatric Gastroenterology and Hepatology, 

Department of Pediatrics, Center for Liver, Digestive, and Metabolic 

Diseases, Beatrix Children’s Hospital - University Medical 

Center Groningen, University of Groningen, Groningen, Netherlands; 

4 Electron Microscopy Center, Jena University Hospital, Jena, 

Germany; 5 Institute of Inorganic and Analytical Chemistry, Friedrich 

Schiller University Jena, Jena, Germany 

Background & Aim: Heme catabolism physiologically produces 

large amounts of bilirubin that can function as potent and regenerative 

scavenger of reactive oxygen species (ROS). Nonetheless, 

ROS attacking heme, biliverdin or bilirubin can lead to the 

formation of higher order metabolites of heme, among them the 

regio-isomeric bilirubin oxidation products (BOXes) Z-BOX A 

and Z-BOX B, with unknown implications for hepatic function. 

The aim of this study was to clarify the appearance of Z-BOX 

A and Z-BOX B under conditions of health and hyperbilirubinemia 

as well as their fate and functional role in the liver as 

a key-player in heme catabolism. Methods: Determination of 

Z-BOX A and Z-BOX B was carried out via HPLC-MS/MS procedures 

in patients suffering from liver failure, healthy human 

controls, Gunn rats as animal model of hereditary bilirubin 

conjugation failure, and Wistar rats. Pharmacokinetics, pharmacodynamics 

and hepatic hemodynamics were studied in 

Wistar rats. Cytoskeletal remodeling, cytotoxicity, and cellular 

metabolism were investigated in cultured HepG2 and HepaRG 

cells by means of fluorescence and scanning electron microscopy 

in addition to biochemical assays. Results: The higher 

order heme metabolites Z-BOX A and Z-BOX B were formed 

under physiological conditions across species (combined levels 

in blood: 25.4 ± 3.4 nmolL -1 , n = 10, healthy human; 

19.1 ± 4.6 nmolL -1 , n = 12, Wistar rat) and were significantly 

elevated under conditions of hyperbilirubinemia, i.e. human 

liver failure (504.3 ± 30.3 nmolL -1 , n = 28; p < 0.0001) and 

animal model of Crigler-Najjar syndrome type I (136.9 ± 15.1 

nmolL -1 , Gunn rat, n = 14; p < 0.0001). Z-BOX A and Z-BOX 

B appeared in an almost fixed ratio (~1.45:1) and highly correlated 

with serum bilirubin (r > 0.86, p = 4.110 19 ). After 

equimolar i.v. bolus application, the pharmacokinetic profiles 

of these regio-isomers exhibited significantly shorter serum half-


life and stronger biliary enrichment (approx. 30x) of Z-BOX A 

compared to Z-BOX B. In parallel, Z-BOX A and Z-BOX B triggered 

an instantaneous and significant reduction of bile flow 

(1 min vs. 3.5 min after bolus injection; n = 5; p = 0.0002). 

Regardless their reported vasoconstrictive properties in cerebral 

arterioles, Z-BOX A and Z-BOX B did not affect hepatic 

hemodynamics. However, Z-BOX A and Z-BOX B showed a 

dose-dependent decrease in cellular metabolic rate (IC 50 

= 

383 mmolL -1 and 141 mmolL -1 , respectively), while only Z-BOX 

A induced a cytoskeletal remodeling of cultured hepatocytes. 

Conclusions: Higher order metabolites of heme are elevated 

under conditions of hyperbilirubinemia and are able to impair 

bile production and cellular integrity. 

Disclosures: 

The following authors have nothing to disclose: Raphael A. Seidel, Franziska 

Schleser, Christoph Sponholz, Frans Cuperus, Sandor Nietzsche, Georg Pohnert, 

Michael Bauer 

809 

NF-kB is critically important for the up-regulation of 

organic solute transporter OSTα/OSTβ in cholestasis 

Natarajan Balasubramaniyan, Frederick J. Suchy; Pediatrics, University 

of Colorado Denver, Aurora, CO 

The up-regulation of the organic solute transporter OSTα/ 

OSTβ in cholestasis provides an alterative pathway for excretion 

of bile acids and other cholephiles across the hepatocyte 

basolateral membrane.However, the mechanisms underlying 

this adaptation have been enigmatic. The mouse and human 

Ostα/β(OSTα/β) promoters contain functional FXR and LRH 

elements, which mediate, respectively, positive and negative 

feedback regulation by bile acids in normal liver, but these 

binding sites don’t explain how this transport system is up-regulated 

in cholestasis when FXR-dependent pathways are compromised.We 

identified previously unknown,well-conserved NF-kB 

binding sites in the Ostα(OSTα) and Ostβ(OSTβ)promoters. 

On chromatin immunoprecipitation(ChIP) analysis of Huh-7 

cells,over-expressed NF-kB was recruited to these sites. In vivo 

ChIP assays done with liver nuclei obtained from mice after 3 

days of CBDL or 6 hours post LPS injection, showed markedly 

increased recruitment of NF-kB p65 to the NF-kB binding sites 

of the Ostα and Ostβ promoters and a diminution of FXR at its 

binding sites (FXRE). Next Huh-7 cells were transiently transfected 

with plasmid vectors containing the OSTα and OSTβ 

promoter sequences that included the NFkB response element 

linked to luciferase as reporter. In contrast to the inhibitory 

effect of NFkB p65 on activity of the BSEP promoter, expression 

of NFkB p65 and p50 subunits together synergistically 

activated the OSTα and OSTβ promoters in a dose-dependent 

fashion. The inhibitors IkBα and IkBSR blocked induction by 

p65 and p50. Previous studies have shown that NF-kB, acting 

as a transcriptional activator, recruits a coactivator complex 

that has striking similarities to that recruited by nuclear receptors 

which usually includes the cyclic AMP response element 

binding protein (CREB)-binding protein (CBP) and p300.CBP/ 

p300 is particularly important in possessing intrinsic histone 

acetyltransferase activity and providing a platform for recruitment 

of a variety of coactivator proteins required for NF-kB-dependent 

gene expression. In vivo ChIP assays using liver nuclei 

obtained from both cholestatic mouse models and an antibody 

that recognizes the homologous CBP and p300 proteins 

showed a markedly increased recruitment of CBP/p300 to the 

NF-kB sites in the Ostα and Ostβ promoters compared with 

controls. In contrast, there was significant depletion of CBP/ 

p300 at the FXRE of Ostα and Ostβ in these models compared 

with controls. These studies reveal a novel, NF-kB-dependent 

mechanism for the up-regulation OSTα/OSTβ in cholestasis, 

providing an alternative export pathway to prevent accumulation 

of hydrophobic bile salts and other toxic products. 

Disclosures: 

The following authors have nothing to disclose: Natarajan Balasubramaniyan, 

Frederick J. Suchy 

810 

The Ileal Bile Acid Transporter Inhibitor A4250 Modulates 

Bile Acid Synthesis and Decreases Serum Bile 

Acids. 

Hanns-Ulrich Marschall 1 , Per-Göran Gillberg 2 , Hans Graffner 2 , Leif 

Rikner 2 ; 1 Department of Molecular and Clinical Medicine, Sahlgrenska 

Academy, Institute of Medicine, University of Gothenburg, 

Gothenburg, Sweden; 2 Albireo, Gothenburg, Sweden 

Background and Aim: Reabsorption of BAs from the intestine 

by ileal bile acid transporter (IBAT) is pivotal for the enterohepatic 

circulation of BAs and sterol homeostasis. We aimed to 

study BA metabolism in a phase 1 trial with the selective IBAT 

inhibitor A4250. Methods: In a double-blind, multiple ascending 

dose design, A4250 capsules or matching placebo was 

administered for 7 days; 1 mg once daily; 3 mg once daily 

and 1.5 mg twice daily, respectively. Each group consisted of 

8 healthy volunteers whereof 6 received active compound and 

2 placebo. BAs were measured by HPLC-MS in plasma and 

feces, FGF19 and C4 in plasma. Results: No serious adverse 

events occurred and all participants finished the trial per protocol. 

Plasma total BAs, and FGF19 decreased, whereas plasma 

C4 and fecal BAs increased. The majority of fecal BAs in the 

A4250 groups were the primary BAs cholic and chenodeoxycholic 

acids. Conclusion: A4250 is a highly efficient and 

well-tolerated compound that by blocking the IBAT in the terminal 

ileum interrupts the enterohepatic circulation of BAs, which 

should be of benefit in patients with cholestatic liver diseases. 

Disclosures: 


Per-Göran Gillberg - Employment: Albireo 

Hans Graffner - Employment: Albireo 

Leif Rikner - Consulting: Albireo AB 

811 

Inhibition of Adenylyl Cyclase 5 reduces cyst growth in 

mice with polycystic liver disease (ADPKD) caused by 

defective Polycystin-2 

Carlo Spirli 1 , Ambra Villani 1 , Valeria Mariotti 1,2 , Luca Fabris 3 , 

Romina Fiorotto 1 , Mario Strazzabosco 1,2 ; 1 Yale University, Section 

of Digestive Diseases, New Haven, CT; 2 Department of Surgery 

and Interdisciplinary Medicine, University of Milano-Bicocca, 

Milan, Italy; 3 Department of Molecular Medicine, University of 

Padova, Padua, Italy 

Genetic defects in Polycystin-1 (PC1) or Polycystin-2 (PC2) 

cause polycystic liver disease associated with ADPKD (PLD). In 

PC-2 defective cystic cholangiocytes, the interaction of STIM1 

(the molecular sensor that mediates Ca 2+ entry following 

ER-Ca 2+ decrease) with ORAI channels is uncoupled accounting 

for a reduced Store Operated Calcium Entry (SOCE) and 

low intracellular and Endoplasmic Reticulum (ER) Ca 2+ levels.


On the other hand, in PC2-defective cells STIM-1 couples with 

a Ca 2+ -inhibitable Adenylyl Cyclase (ACs) and stimulates the 

store-operated production of cAMP (SOcAMP), PKA-dependent 

activation of the ERK1/2 and increase in VEGF production 

by cystic cells. This mechanism plays a key role in promoting 

liver cyst growth in PLC. Two Ca 2+ -inhibitable ACs (AC6 and 

AC5) are expressed in cholangiocytes and have similar properties, 

however the molecular identity of the AC responsible for 

inappropriate cAMP production in PLC is unknown. Results: 

PC2-conditional KO mouse (Pkd2 flox/- ;pCxreER TM ) (Pkd2cKO) 

were crossed with AC6 -/- mouse to generate double Pkd2/ 

AC6KO mice. Successful deletion was confirmed by RT/PCR. 

Mice were sacrificed eight weeks after induction of PC2 excision 

and cystic area was assessed by K19 staining and computer-assisted 

morphometric analysis. However, Pkd2/AC6KO 

mice did not show a reduction of the liver cystic area as respect 

to Pkd2cKO mice. Moreover, in cystic cholangiocytes isolated 

from Pkd2/AC6KO mice, intracellular levels of cAMP generated 

after an acute ER [Ca 2+ ] depletion (TPEN 1mM) were not 

different from those measured in cholangiocytes from Pkd2cKO 

mice, suggesting that in absence of AC6, the AC5 isoform, may 

be activated. In fact, inhibition of AC5 by two different inhibitors 

(NKY90 and SQ 22,536) significantly reduced cAMP 

following ER [Ca 2+ ] depletion. Consistent with these data, in 

vivo treatment of Pkd2/AC6KO with SQ 22,536 ((300 mg/Kg 

day, I.P) caused a significant reduction in liver cystic area was 

significantly reduced, along with the liver/body weight ratio. 

In conclusion, these data indicate that AC5 is the Ca2+ inhibitable 

AC responsible for cAMP production by PC2-defective 

cells in condition of low cellular and ER [Ca 2+ ] and the mechanisms 

is of pathophysiological relevance. Inhibition of cAMP 

production by somatostatin analogs has been shown to be of 

therapeutic value in patients with PLC. Our study indicate that 

pharmacologic inhibition of AC5 may represent a more specific 

target to reduce of cyst growth in polycystic liver diseases. 

Disclosures: 

The following authors have nothing to disclose: Carlo Spirli, Ambra Villani, Valeria 

Mariotti, Luca Fabris, Romina Fiorotto, Mario Strazzabosco 

812 

Cholestatic-induced biliary proliferation and fibrosis 

are decreased in mice lacking mast cells: an innovative 

study using C-kit knockout mice 

Laura Hargrove 1 , Lindsey Kennedy 2 , Jennifer Owens 2 , Heather 

L. Francis 2,1 ; 1 Scott and White Memorial Hospital, Temple, TX; 

2 Research, Central Texas Veterans Health Care System, Temple, TX 

Background: Cholestatic liver injury is marked by increased 

biliary proliferation. Bile duct ligation (BDL) induces large, but 

not small, cholangiocyte proliferation coupled with increased 

fibrosis. Mast cells (MCs) are inflammatory cells that, once 

activated, release histamine (HA). Our previous studies have 

demonstrated that HA increases vascular endothelial growth 

factor (VEGF) expression and knockdown of histidine decarboxylase 

(HDC (synthesizes histamine)) decreases biliary VEGF 

expression/secretion. We have demonstrated that MCs infiltrate 

the liver following BDL increasing intrahepatic bile duct 

mass (IBDM) and fibrosis. Inhibition of MC-derived HA using 

cromolyn sodium decreases these parameters suggesting that 

MCs contribute to biliary proliferation and fibrosis. Our aim 

was to evaluate the effects of BDL on biliary proliferation and 

fibrosis in MC deficient mice. Methods: WT and C-kit -/- mice 

(MC deficient) were subjected to sham or BDL for 3 days before 

collecting serum, liver blocks and pure cholangiocytes. IBDM 

and proliferation were evaluated in sections by CK-19 and 

PCNA immunohistochemistry, respectively. In total liver (TL) 

and cholangiocytes we evaluated PCNA expression by qPCR 

and western blotting. Apoptosis was measured by gene expression 

for Bax and BCL-2 and TUNEL staining in tissues. Fibrosis 

was detected by Sirius Red/Fast Green staining in tissues and 

by qPCR for α-SMA, fibronectin, collagen type-1a and TGFβ1 

in TL. In cholangiocytes and TL we measured HDC and 

VEGF-A/C, gene expression and secretion by EIA in serum 

and cholangiocyte supernatants. To determine if MC-derived 

HA regulates biliary proliferation and fibrosis in vitro, cultured 

MCs were transfected with HDC shRNA prior to co-culture with 

murine cholangiocytes. Biliary proliferation was measured by 

MTS, and VEGF expression and fibrosis marker expression by 

qPCR. Results: In BDL C-kit -/- mice there was enhanced ductular 

reaction with increased small IBDM and decreased large IBDM 

compared to BDL WT. Proliferation and liver fibrosis were 

decreased in BDL C-kit -/- mice compared to BDL WT coupled 

with enhanced apoptosis. HDC and VEGF-A/C expression was 

decreased in BDL C-kit -/- mice as was HA and VEGF secretion 

in serum and supernatants compared to BDL WT. In vitro, 

knockdown of MC-HDC expression decreased biliary proliferation, 

VEGF expression and fibrosis suggesting MC-derived 

HA contributes to biliary proliferation and fibrosis. Conclusion: 

BDL-induced biliary proliferation and fibrosis are decreased in 

mice lacking mast cells via the HDC/HA/VEGF axis. Our studies 

support the novel concept that hepatic mast cells are critical 

to biliary response following injury. 

Disclosures: 

The following authors have nothing to disclose: Laura Hargrove, Lindsey Kennedy, 

Jennifer Owens, Heather L. Francis 

813 

YAP Reduces Liver Parenchymal Damage by Promoting 

Bile Duct Capacity in Cholestatic Injury 

Quy P. Nguyen 1 , Nan Wu 2 , Tianhao Zhou 2 , Haibo Bai 1 ; 1 BaylorScott&White 

Hospital, Temple, TX; 2 Texas A&M University College 

of Medicine, Temple, TX 

Background&Aims: YAP is highly expressed in cholangiocytes. 

YAP protein levels are further increased in cholangiocytes after 

bile duct ligation (BDL). However, the physiological role of 

biliary YAP in maintaining biliary homeostasis and in response 

to cholestatic injury is not clear due to lacking efficient methods 

to ablate YAP in cholangiocytes. Methods: We found that overexpression 

of YAP in hepatocytes induces a compensatory loss 

of YAP in cholangiocytes. Moreover, even suppressing YAP 

activity with transgenic Vgl4, a YAP competitor, is not able to 

rescue the YAP deficiency in cholangiocytes. Because Yap/ 

Vgl4 double transgenic mice (Yap/Vgl4 DTg) have normal 

liver function, we use it as model mice to study the function of 

YAP in cholangiocytes. Sham or BDL was performed with these 

groups of mice. Two weeks later, liver histology, double immunofluorescence 

staining (E-cadherin and CK19), immunohistochemistry 

staining (CK19), Sirius red staining were compared 

between control and Yap/Vgl4 DTg groups. Primary cholangiocytes 

were isolated from CK19-CreER; Yap flox/flox mice and 

cultured in vitro. Yap deletion was induced through addion of 

4-OH tamoxifen. Immunofluorescence staining of E-cadherin 

was compared between control and Yap KO cholangiocytes. 

YAP expression in bile ducts of the Mdr2 -/- mouse model of 

primary sclerosing cholangities was also studied by immunohistochemistry. 

Results: Bile infarcts and fibrosis were more 

severe in Yap/Vgl4 DTg mice compared to their WT littermate 

controls. CK19 staining revealed bile ducts are more dilated 

in WT controls than in Yap/Vgl4 DTg livers. E-cadherin staining 

revealed more well formed E-cadherin junction between 

cholangiocytes in Yap/Vgl4 DTg mice. E-cadherin junctions


are also more mature in primary cholangiocytes with Yap deficiency. 

YAP activity correlates with Mdr2-/- mice disease progress. 

Conclusions: YAP functions to increase bile duct dilation 

upon bile duct obstruction through promoting cholangiocyte 

E-cadherin junction dynamics. The dilated bile ducts are able 

to hold more bile in bile ducts of WT controls, which in turn 

reduces hepatocyte damage caused by bile overflow. 

Disclosures: 

The following authors have nothing to disclose: Quy P. Nguyen, Nan Wu, Tianhao 

Zhou, Haibo Bai 

814 

Glutathione depletion is critical for the early pathogenesis 

of biliary atresia (BA) in the toxin-induced BA model 

Xiao Zhao 1 , Benjamin J. Wilkins 2 , Kristin Lorent 1 , John R. Porter 3 , 

Rebecca G. Wells 1 , Michael Pack 1 ; 1 Medicine/Division of Gastroenterology, 

University of Pennsylvania, Philadelphia, PA; 2 Division 

of Anatomic Pathology, Department of Pathology and Laboratory 

Medicine, The Children’s Hospital of Philadelphia, Philadelphia, 

PA; 3 University of the Sciences, Philadelphia, PA 

Biliary atresia (BA) is a complex neonatal cholangiopathy that 

is the leading indication for liver transplantation in the pediatric 

population. Using an in vivo zebrafish biliary assay, we have 

isolated biliatresone, a novel plant isoflavonoid with selective 

extrahepatic biliary toxicity that is responsible for BA outbreaks 

in newborn Australian livestock. The phenotype induced in 

this new BA model is conserved between zebrafish and mammals, 

and recapitulates the cardinal features of human BA. 

While extrahepatic cholangiocytes are exquisitely sensitive to 

biliatresone, hepatocytes and intrahepatic cholangiocytes are 

resistant to its deleterious effect. This difference in susceptibility 

can potentially be explained by the intrinsic differential capability 

of specific liver cell types in mitigating stress. Global 

expression profiling of mRNA recovered from larval zebrafish 

cholangiocytes and total liver revealed that biliatresone 

leads to early up-regulation of the glutathione (GSH) redox 

response, suggesting that GSH depletion is an inciting event 

in biliatresone-mediated biliary injury. GSH, as a ubiquitous 

tripeptide thiol, is a vital cellular antioxidant. These data are 

consistent with in vitro assays that confirm the reactivity of biliatresone 

with glutathione. Using a specific genetically encoded 

GSH redox probe (redox-sensitive GFP, roGFP), we were able 

to establish quantitative in vivo mapping of the GSH redox 

potential (E GSH 

) in hepatocytes and extrahepatic cholangiocyte 

(EHC) of the biosensor-transgenic larvae. Interestingly, we 

detected endogenous differences in the redox status between 

hepatocytes and EHC. Pharmacological manipulation of GSH 

redox homeostasis further supported the importance of GSH in 

modulating biliatresone-induced injury. This is evidenced by the 

temporary attenuation of its biliary toxicity with N-acetylcysteine 

(NAC), a GSH precursor. Altogether, these data strongly 

suggest redox stress as a contributing factor in biliatresone-induced 

injury and differences in intrinsic stress responses can 

explain its cell-type specificity. Insufficient antioxidant capacity 

of EHC may potentially be critical to the early pathogenesis of 

BA. 

Disclosures: 

The following authors have nothing to disclose: Xiao Zhao, Benjamin J. Wilkins, 

Kristin Lorent, John R. Porter, Rebecca G. Wells, Michael Pack 

815 

The bile acid Ursodeoxycholic acid activates cholangiocyte 

TMEM16A Cl- channels 

Qin Li, Amal K. Dutta, Charles Kresge, Andrew P. Feranchak; 

Department of Pediatric Gastronenterology, Hepatology, University 

of Texas Southwestern Medical Center, Dallas, TX 

Ursodeoxycholic acid (UDCA) stimulates a bicarbonate-rich 

choleresis in part through effects on cholangiocytes. During 

cholehepatic shunting, uptake of bile acids at the apical cholangiocyte 

membrane is associated with an increase in [Ca2+] 

I, Cl- efflux and Cl-/HCO3- exchange, though the cellular 

mechanism is unknown. As TMEM16A is a Ca2+-activated Clchannel 

in the apical membrane of cholangiocytes (JBC 2011; 

286:766-776) the aim of the present study was to determine if 

TMEM16A is the target of UDCA-stimulated Cl- secretion and 

to identify the regulatory pathway involved. Methods: Studies 

were performed in SV40-immortalized mouse cholangiocytes 

isolated from the large intrahepatic ducts (MLC) and in vivo 

studies were performed in a novel bile duct-cannulated murine 

model which we recently developed. This model involves placement 

of an indwelling catheter into the bile duct of a live mouse 

for measurement of bile flow rate and composition in response 

to systemic administration of agonists. [Ca2+]I was measured 

by Fura-2, ATP release by luciferase based assay and reported 

as arbitrary light units (ALUs), and Cl- currents by patch clamp 

techniques. Results: Exposure of MLC to UDCA (100 mM) rapidly 

increased [Ca2+]I (increase in Fura-2 fluorescence from 

0.65 to 1.05, p ≤0.01), stimulated ATP release (from 331 ± 22 

ALUs to 780 ± 106 ALUs, p


816 

The bile-acid receptor TGR5 regulates biliary epithelium 

permeability 

José Ursic-Bedoya 1,2 , Hayat Simerabet 1,2 , Isabelle Doignon 1,2 , 

Noémie Péan 1,2 , Zahra Tanfin 1,2 , Christoph Ullmer 3 , Lydie Humbert 

4,5 , Dominique Rainteau 4,5 , Doris Cassio 1,2 , Thierry Tordjmann 

1,2 ; 1 U1174, INSERM, Orsay, France; 2 Université Paris sud, 

Orsay, France; 3 Pharma Research and Early Development, Roche 

Innovation Center Basel, Basel, Switzerland; 4 U1057, INSERM, 

Paris, France; 5 UPMC, Paris, France 

Background: TGR5, the bile acid (BA) G-protein-coupled receptor 

protects the liver against BA overload during regeneration. 

After partial hepatectomy, peribiliary hepatic necrosis occurs 

in TGR5 KO but not in wild type (WT) mice, suggesting that 

TGR5, highly expressed in biliary epithelial cells, regulates 

biliary epithelium permeability. Methods: Trans-epithelial resistance 

(TER) and FITC-dextran transfer were measured in the 

NRC (Normal Rat Cholangiocyte) cell line cultured on transwell 

inserts. To study biliary epithelial permeability in vivo in mice, 

fluorescent dextran or a modified BA (Glycocholic acid) were 

injected gallbladder (GB) lumen and traced (spectrofluorimetry 

& Mass Spectrometry) in plasma and liver. Cells and mice 

were stimulated with RO5527239, a TGR5 specific agonist, 

and with taurolitocholic acid. TGR5-induced signaling pathways 

were studied by western blot (WB), using selective inhibitors. 

Tight junction (TJ) proteins expression was investigated by 

qPCR, WB and immunofluorescence in NRC, in livers and GB 

from WT and TGR5-KO mice. Results: In NRC, TGR5 agonists 

significantly increased TER, reduced dextran passage, induced 

ERK phosphorylation and EGFR transactivation. Inhibition of 

cAMP production or MAPK pathways suppressed TGR5 agonists 

effect on NRC permeability. In TGR5-KO as compared 

with WT mice, although TJ proteins (ZO-1, occludin and JAM- 

A) mRNA expression in GB was significantly reduced, only 

JAM-A expression and localization at TJ were altered in bile 

ducts and GB. TGR5 agonists induced JAM-A phosphorylation 

and TJ localization in vitro in NRC, and in vivo after injection 

in WT but not in TGR5-KO GB lumen. In vivo, BA and dextran 

transepithelial transfer after GB injection was increased in 

TGR5-KO as compared with WT mice (Fig.1). BA transporters 

mRNA expression (ASBT, OSTb, MRP3) was similar in WT 

and TGR5-KO GB, suggesting that TGR5 controls paracellular 

rather than transcellular permeability. Conclusion: The BA 

receptor TGR5 reinforces biliary epithelial sealing in vitro and 

in vivo, likely through modulation of TJ protein expression and 

phosphorylation, protecting liver parenchyma against bile leakage. 

Disclosures: 

Christoph Ullmer - Employment: F. Hoffmann-La Roche AG 

The following authors have nothing to disclose: José Ursic-Bedoya, Hayat Simerabet, 

Isabelle Doignon, Noémie Péan, Zahra Tanfin, Lydie Humbert, Dominique 

Rainteau, Doris Cassio, Thierry Tordjmann 

817 

Knockout of the Secretin/Secretin Receptor Axis Delays 

Regeneration of small and large cholangiocytes by 

enhanced senescence following 70% partial hepatectomy 

(PH) 

Ying Wan 3 , Shannon S. Glaser 1 , Nan Wu 4 , Fanyin Meng 2 , 

Julie Venter 4 , Romina Mancinelli 5 , Heather L. Francis 2 , Antonio 

Franchitto 5 , Paolo Onori 5 , Tina Kyritsi 4 , Shanika Avila 4 , Guido 

Carpino 7 , Holly A. Standeford 6 , Gianfranco Alpini 2 , Eugenio 

Gaudio 5 ; 1 Central Texas Veterans Health Care System and Texas 

A&M HSC College of Medicine, Temple, TX; 2 Research, S&W 

DDRC and Medicine, Veterans Health Care System and Texas 

A&M HSC and BaylorScott&White, Temple, TX; 3 Operational 

Funds, BaylorScott&White, Temple, TX; 4 Medicine, Texas A&M 

University HSC, Temple, TX; 5 Department of Anatomical, Histological, 

Forensic Medicine and Orthopedics Sciences, University La 

Sapienza, Rome, Italy; 6 Research, Central Texas Veterans Health 

Care System, Temple, TX; 7 Dept Health Sciences, University of 

Rome “Foro Italico, Rome, Italy 

Limited data exists regarding the neuroendocrine factors that 

regulate the renewal of the biliary tree after PH. The secretin 

(SCT)/secretin receptor (SR) axis is normally expressed only by 

large cholangiocytes. However, during the damage of large 

bile ducts small cholangiocytes (more resistant to injury) replenish 

the large damaged ducts by de novo acquisition of large 

biliary phenotypes. Cellular senescence is a state of irreversible 

cell cycle arrest involved in biliary diseases including primary 

sclerosing cholangitis and primary biliary cirrhosis. Large, 

senescent cholangiocytes are the target cells in the cholestatic 

models of bile duct ligation and MDR2 KO (PSC). Hypothesis: 

in hepatectomized SCT/SR double knockout (DKO) mice there 

is reduced regeneration of small and large bile ducts due to 

enhanced senescence. Methods: The studies were performed 

in normal wild type (WT) and SCT/SR DKO mice at 0, 3, 6 

hours and 1, 3, 7, and 14 days after sham or 70% PH. Liver 

injury was evaluated by: (i) H&E staining in liver sections; and 

(ii) measurement of serum levels of transaminases. We evaluated 

biliary proliferation by: (i) immunohistochemistry (IHC) for 

PCNA and intrahepatic bile duct mass (IBDM) by CK-19 IHC in 

liver sections; and (ii) qPCR for PCNA and CK-19 in total liver 

samples and small and large cholangiocytes. Cellular senescence 

was evaluated by SA-β-Gal staining in liver sections 

and qPCR for P18, PAI-1 and CCl2 in total liver samples and 

small and large cholangiocytes. Results: Small cholangiocytes 

from PH WT mice displayed less senescent features compared 

to large cholangiocytes. In PH WT mice, small less senescent 

cholangiocytes regenerate as early as 1 hr and rebuild IBDM 

within 1 day of PH, whereas large regenerate as early as 6 hr 

and rebuild IBDM at a lower rate (after 3-7 days of PH). Large 

cholangiocytes from DKO PH animals displayed increased 

expression of senescent markers relative to PH WT mice. In 

DKO PH mice, there was reduced regeneration time (by PCNA) 

and regrowth (by CK-19) of small and large bile ducts concomitant 

with enhanced expression of senescence markers (p18, 

PAI-1 and CCL2) in large cholangiocytes as well as in small 

cholangiocytes that display normally low levels of senescence. 

In SCT/SR double KO mice, at day 14 of PH IBDM returned at 

only 50% of the original biliary mass (0.238 ± 0.014, sham, 

vs. 0.128 ± 0.007, SCT/SR KO, p


Disclosures: 

The following authors have nothing to disclose: Ying Wan, Shannon S. Glaser, 

Nan Wu, Fanyin Meng, Julie Venter, Romina Mancinelli, Heather L. Francis, 

Antonio Franchitto, Paolo Onori, Tina Kyritsi, Shanika Avila, Guido Carpino, 

Holly A. Standeford, Gianfranco Alpini, Eugenio Gaudio 

818 

Pre-treatment with TIMP-3 prevents the development of 

biliary injury in an LPS enhanced ischaemia-reperfusion 

animal model 

Janske Reiling 1,2 , Kim Bridle 1,3 , Catherine M. Campbell 4 , Nishreen 

Santrampurwala 1,3 , Laurence J. Britton 1,3 , Ari J. Cohen 5 , Darrell H. 

Crawford 1,3 , Cornelis H. Dejong 2,6 , Jonathan Fawcett 1,7 ; 1 school 

of medicine, University of Queensland, Brisbane, QLD, Australia; 

2 NUTRIM School of Nutrition and Translational Research in Metabolism, 

Maastricht University, Maastricht, Netherlands; 3 Gallipoli 

Medical Reserach Foundation, Brisbane, QLD, Australia; 4 Envoi 

Specialist Pathologists, Brisbane, QLD, Australia; 5 Laboratory of 

Transplant Reseach, Institute of Translational Research, Ochsner 

Clinical Foundation, New Orleans, LA; 6 Department of Surgery, 

Maastricht University Medical Centre, Maastricht, Netherlands; 

7 Queensland Liver Transplant Service, Princess Alexandra Hospital, 

Brisbane, QLD, Australia 

Introduction Transplantation of donor livers retrieved after circulatory 

death is often complicated by the development of biliary 

injury and subsequent stricture formation. Our recent studies 

identified a role for lipopolysaccharides (LPS) in the development 

of biliary injury. This study aimed to assess whether 

inhibition of TNF-α cleavage into its biologically active form, 

by pre-treatment with tissue inhibitor of metalloproteinases-3 

(TIMP-3), would prevent the development of biliary injury. 

Methods Male Sprague Dawley rats underwent either sham 

surgery (sham), 1000ng/kg human recombinant TIMP-3 one 

hour prior to sham surgery (TIMP-3), 70% partial liver ischaemia 

for thirty minutes with 1mg/kg LPS (combi) or TIMP-3 

followed by ischaemia and LPS (combi+TIMP). Following 6 

hours of reperfusion, blood, bile, liver and bile duct tissue was 

collected for analysis of liver function, biliary injury and expression 

of genes encoding for bile acid transporters and cytokines. 

Results TIMP-3 treatment prior to liver ischaemia and LPS administration 

resulted in a decreased bile duct injury severity score 

compared to non-pre-treated animals (sham: 0.00 (0.00-3.00); 

TIMP: 0.00 (0.00–4.00); combi: 4.50 (1.00-6.00); combi+TIMP: 

2.00 (0.00–5.00)). This was further supported by a 

significant decrease in lactate dehydrogenase in bile, a marker 

of biliary injury (combi: 15.78±9.46; combi+TIMP: 6.54±2.33 

U/L/ gram wet liver weight p=0.007). Despite improvement 

of biliary injury, hepatocellular injury remained unaltered as 

evidenced by increased serum alanine transaminase in both 

the combi and combi+TIMP groups (sham: 40.90±9.63; 

combi: 109.64±40.96; combi+TIMP: 114.63±84.23, p: 

0.03 and 0.024 respectively). Bile flow was normalised following 

TIMP pre-treatment (combi: 64.20±14.57; combi+LPS: 

85.37±10.99, p=0.012), as were gene expression levels of 

CYP27a1 and CYP7b1 (2 fold and 1,5 fold reduction, p=0.01 

and 0.02 respectively). This may reflect a normalisation of total 

bile acid synthetic capacity. Furthermore, gene expression of 

MIP-2, osteopontin and LBP were significantly down regulated 

following TIMP pre-treatment (8 fold, 6 fold and 2 fold respectively, 

p


oncocytic type were common. In contrast, group C and D were 

located in perihilar and distal bile ducts, and almost all cases 

were of high grade with invasion, and a majority of them were 

of intestinal and pancreatobiliary phenotypes. Group B was 

found along the biliary tree, and a majority of cases were of 

intestinal phenotype and all were of high grade with or without 

invasion. Conclusion: PNB with histopathologic similarities to 

IPMN and those without similarities to IPMN were different in 

their location and pathological behaviors. IPNB may be a heterogeneous 

group showing different biological behaviors and 

undergoing different tumorigenesis. 

Disclosures: 

The following authors have nothing to disclose: Yasuni Nakanuma, Yuko Kakuda, 

Katsuhiko Uesaka, Takashi Miyata, Yuki Fukumura, Masaru Takase 

Disclosures: 

The following authors have nothing to disclose: Natalie L. Berntsen, Bjarte Fosby, 

Corey Tan, Elisabeth Schrumpf, Tonje Bjoernetroe, Aksel Foss, Pål-Dag Line, Tom 

H. Karlsen, Richard S. Blumberg, Espen Melum 

820 

Intraductal papillary neoplasm of bile duct is a heterogeneous 

disease with respect to its histopathologic 

similarities to pancreatic intraductal papillary mucinous 

neoplasm 

Yasuni Nakanuma 1 , Yuko Kakuda 1 , Katsuhiko Uesaka 1 , Takashi 

Miyata 1 , Yuki Fukumura 2 , Masaru Takase 3 ; 1 Shizuoka Cancer 

Center, Sunto-Nagaizumi, Japan; 2 Department of Pathology, Juntendo 

University School of Medicine, Tokyo, Japan; 3 Department 

of Clinical Laboratory, Koshigawa City Hospital, Koshigaya, Japan 

Background and Aim: Intraductal papillary neoplasm of bile 

duct (IPNB) is characterized by papillary tumor covered by 

well-differentiated neoplastic epithelium with fine fibrovascular 

cores in the dilated bile ducts. IPNB reportedly resembles 

intraductal papillary mucinous neoplasm (IPMN) of pancreas 

to a varied degree, though the exact comparison remain to be 

clarified. Herein, we examined IPNB with respect to its histopathologic 

similarities to IPMN. Materials and Methods: A total 

of surgically resected 52 cases of IPNB and 42 cases of IPMN 

(with mural nodule) which met the WHO 2010 criteria of IPNB 

and IPMN were histologically, immunohistochemically and 

semiquantitatively compared with respect to the size, grade 

of intraepithelial neoplasm (low/intermediate, and high grade 

without invasion and with invasion), mucus hypersecretion, 

and phenotypes (intestinal, gastric, pancreatobiliary and oncocytic 

type) with a help of immunostaining of MUC1, MUC2, 

MUC5AC, MC6, CK7, CK20, CDX2 and CD10. This comparative 

study was done by two experts of biliary pathology and 

those of pancreatic pathology. Results: There were no differences 

in gender and age distributionof IPNB and IPMN (F:M 

35:17 in IPNB and 26:16 in IPMN, mean age 70.1 yrs in IPNB 

and 67.1 in IPMN). Mucus hypersecretion was more freuqent 

in IPMN (83%) than in IPNB (39%). Grades were different in 

IPNB and IPMN; low/intermediate (9.6% and 38.1%), high 

grade (25% and 23.8%) and high grade with invasion (65.4% 

and 38.1%). IPNB cases were classifiable into four groups with 

their histopathological similarities to IPMN: group A (identical 

to IPMN, 19 cases), group B (similar but a little different from 

IPMN, 18 cases), group C (focally or vaguely similar to IPMN, 

5 cases), and group D (different from IPMN, 10 cases). Group 

A was mainly found in the intrahepatic and perihilar bile ducts, 

and a majority of them were low/intermediate grade or high 

grade intraepithelial neoplasm without invasion. Gastric and 

821 

Wnt/β-catenin cooperates with Yap signaling to promote 

hepatobiliary repair in a model of chronic liver 

injury 

Kari Nejak-Bowen, Hirohisa Okabe, Satdarshan (Paul) S. Monga; 


Sustained cholestasis results in injury to the biliary epithelium 

with subsequent atypical ductular proliferation (ADP), which 

is thought to be a reparative mechanism. ADP is observed in 

animal models where biliary injury is the primary insult to the 

liver, such as 0.1% 3,5-diethoxycarbonyl-1,4-dihydro-collidine 

(DDC). Intriguingly, during cholestatic injury, small subsets of 

hepatocytes also begin to acquire a cholangiocyte-like phenotype. 

This phenomenon, known as transdifferentiation, or 

cellular reprogramming, may contribute to biliary repair by 

creating de novo channels for bile export. Because transgenic 

mice expressing a serine 45-mutated stable form of β-catenin 

(S45-TG) in liver have a significant number of hepatocytes 

expressing A6 (a biliary marker), after long-term exposure to 

DDC compared to wild-type (WT) littermates, we hypothesized 

that Wnt signaling plays a role in this process. To test this, 

WT animals on control and DDC diet were analyzed for Wnt 

expression. We found upregulation of Wnt 7a, 7b, and 10a 

– in the portal triad of DDC-fed mice, as determined by tissue 

microdissection. Cell sorting showed expression of these Wnts 

exclusively in the EpCAM+ cell compartment. Direct co-culture 

of Hek293 cells expressing Wnt7a significantly increased 

Sox-9 gene expression in AML12 cells and primary hepatocytes. 

Treatment with recombinant Wnt7A protein also induced 

Sox9 and EpCAM expression in AML12 cells. We further 

analyzed livers of WT and S45-TG mice on DDC diet and 

observed increased expression of biliary markers EpCAM and 

cytokeratin 19 (CK19) in S45-TG mice, which occurs as early 

as 28d after DDC. Concurrently, S45-TG hepatocytes in the 

periportal region also begin to express Sox9, a transcription 

factor normally restricted to bile ducts. Additionally, mice that 

lack Wnt signaling in liver have fewer A6-positive hepatocytes 

after bile duct ligation, further implicating Wnt signaling in this 

process. Finally, Yap activation, which regulates liver cell fate, 

was notably increased in hepatocyte nuclei of S45-TG livers, 

concomitant with increased expression of Yap target genes. 

Thus, Wnt/β-catenin signaling cooperates with the Yap pathway 

to regulate liver repair after biliary injuries. 

Disclosures: 

Satdarshan (Paul) S. Monga - Advisory Committees or Review Panels: Abbvie; 

Grant/Research Support: Dicerna Pharmaceuticals 

The following authors have nothing to disclose: Kari Nejak-Bowen, Hirohisa 

Okabe


822 

Prolonged usage of H1 or H2 histamine receptor antagonists 

decreases fibrosis and liver damage in MDR2 - 

/- 

mice: novel evidence of the therapeutic benefits of 

anti-histamines 

Hannah Jones 1 , Laura Hargrove 1 , Lindsey Kennedy 2 , Jennifer 

Owens 2 , Heather L. Francis 2,1 ; 1 Scott and White Memorial Hospital, 

Temple, TX; 2 Research, Central Texas Veterans Health Care 

System, Temple, TX 

Background: Primary Sclerosing Cholangitis (PSC) arises from 

damaged cholangiocytes, epithelial cells lining the biliary 

epithelium. The multi-drug resistant 2-gene knockout mouse 

(MDR2 -/- ) is similar to human PSC. Histamine increases biliary 

proliferation and intrahepatic ductal mass (IBDM) by 

interaction with histamine receptors (HRs). H1HR signals via 

increased PKC/Ca 2+ signaling, whereas H2HR induces effects 

via cAMP-dependent signaling. H1/H2 HR blockers like Claritin® 

and Zantac® are used to alleviate allergic reactions and 

heartburn. Our current study aimed to evaluate the effects of 

prolonged usage of H1 and H2 blockers on biliary proliferation, 

hepatic damage and fibrosis in MDR2 -/- mice. Methods: 

Wild-type (WT) and MDR2 -/- mice were treated by osmotic 

minipumps with 0.9% NaCl, fexofenadine (10mg/kg BW/ 

day) or ranitidine (10mg/kg BW/day) for 4 weeks to recapitulate 

long-term usage. Liver blocks, serum and isolated cholangiocytes 

were obtained. Cholangiocyte proliferation and IBDM 

was evaluated by qPCR for PCNA and immunohistochemistry 

for CK-19 (bile duct marker). Fibrosis was detected by Masson’s 

Trichrome staining and by qPCR for the fibrotic markers, 

α-SMA, fibronectin, collagen type-1a and TGF-β1. In serum, 

ALT and AST levels were measured and histamine and TGF-β1 

by EIA. Necrosis, inflammation, and lobular damage were 

assessed by H&E. Biliary function was measured in purified 

cholangiocytes by evaluating intracellular IP 3 

and cAMP levels 

by EIA. In vitro, cholangiocytes were treated with fexofenadine 

or ranitidine (25 μM) for up to 72 hours before measuring 

biliary proliferation by MTS assay and PCNA immunoblots 

and the expression of the aforementioned fibrotic markers by 

qPCR. Results: In MDR2 -/- mice treated with H1/H2 HR blockers 

there was a significant decrease in liver damage, IBDM, 

proliferation and fibrosis compared to saline treated MDR2 - 

/- 

mice. AST and ALT levels were decreased in mice treated 

with HR blockers and histamine and TGF-β1 secretion was 

also reduced compared to saline treated MDR2 -/- mice. Also, 

IP 3 

and cAMP levels were decreased in cholangiocytes from 

MDR2 -/- mice treated with H1 or H2 blockers, respectively. No 

adverse effects on biliary proliferation, liver damage or fibrosis 

were found in WT mice treated with HR blockers. In vitro, stimulation 

with H1 or H2 blockers reduced biliary proliferation via 

IP 3 

/Ca 2+ or cAMP signaling, respectively. Conclusion: Chronic 

usage of H1 or H2 blockers has no adverse effect on WT mice, 

but reduces liver damage, biliary proliferation and fibrosis in 

MDR2 -/- mice. These drugs should be considered as a promising 

therapy for patients suffering from cholangiopathies such 

as PSC. 

Disclosures: 

The following authors have nothing to disclose: Hannah Jones, Laura Hargrove, 

Lindsey Kennedy, Jennifer Owens, Heather L. Francis 

823 

Hepatic CD4+ lymphocyte responses and initiation of 

biliary injury in mdr2 knockout mice depend on dendritic 

cell costimulation 

Celine S. Lages 1 , Tiffany Shi 1 , Paige Bolcas 1 , Julia Simmons 1 , 

Avery Maddox 1 , Shiva K. Shanmukhappa 2 , Alexander G. 

Miethke 1 ; 1 Department of Gastroenterology, Cincinnati Children’s 

Hospital Medical Center, Cincinnati, OH; 2 Pathology and Laboratory 

Medicine, Cincinnati Children’s Hospital Medical Center, 

Cincinnati, OH 

T lymphocytes are implicated in initiation of cholangiopathies 

like biliary atresia or primary sclerosing cholangitis (PSC). 

CD11c+ dendritic cells (DCs) modulate expansion and activation 

of T lymphocytes through CD80/86-mediated costimulation. 

Blockade of CD86/CD28 at the immunological synapse 

with the CTLA4-IgG fusion protein was shown to improve autoimmune 

diseases. We hypothesize that DC costimulation is 

critical for T lymphocyte activation during the initiation of bile 

duct epithelial injury in mdr2-/- mice, a model for “toxic bile” 

induced sclerosing cholangitis. Methods: Kinetics of hepatic 

DC and T cell responses in male double-transgenic mdr2 -/- 

and CD11c-GFP mice were determined by flow cytometry. For 

costimulatory blockade, mice received i.p. injections of 20 

mcg/g of CTLA4-Ig every other day between day of life 7 

and 15. Plasma ALT levels and ductal proliferation on CK19- 

stained liver sections served as surrogate markers for hepatocellular 

and biliary injury, respectively. Results: Frequency of 

hepatic CD11c (GFP)+ DCs was increased in mdr2-/- mice 

at d8 and d14, but decreased thereafter (mean % GFP+MH- 

CII+B220-/Lin-7AAD- at d8: 0.46 vs 0.27, p=0.08; at d14: 

0.96 vs 0.46, p=0.02; at d30: 1.30 vs 2.95, p=0.03 in 

mdr2-/- vs mdr2+/+). The immunogenic subset of CD11b+ 

myeloid mDCs followed a similar kinetic (d8: 0.22 vs 0.09, 

p=0.02; d14: 0.77 vs 0.12, p=0.001; d30: 1.12 vs 2.52, 

p=0.04). Frequency of DCs expressing the costimulatory molecules 

CD80 or CD86 were increased at d8 and d14, but 

decreased at d30. Treatment of mdr2-/- mice with CTLA4-Ig 

was associated with significantly decreased plasma ALT levels 

at d15 (mean 162.1 vs 465.1 IU/L in CTLA4-Ig vs non-treated, 

p=0.02) and reduced number of bile duct profiles per portal 

tract (mean 1.51 vs 2.11 in CTLA4-Ig vs hIgG, p=0.002). 

Administration of CTLA4-Ig was associated with reduced 

hepatic CD4+ T cell responses (mean % CD4+CD3+/7AAD-: 

2.37 vs 3.94, p=0.02), but did not affect expansion of CD8+ 

T, NK or NKT cells. Typical for costimulatory blockade, differentiation 

of naïve to effector CD4+ T cells was attenuated 

(mean % of hepatic effector memory CD62L-CD44+/CD4: 

10.82 vs 60.90, p


824 

Tetrahydroxylated Bile Acids Reduce Cholangiocyte 

Damage in Abcb4 -/- mice 

Renxue Wang 1 , Jonathan Sheps 1 , Lin Liu 1 , Victor Ling 1,2 ; 1 BC Cancer 

Research Centre, Vancouver, BC, Canada; 2 Pathology, University 

of British Columbia, Vancouver, BC, Canada 

Background and Aim: Primary sclerosing cholangitis (PSC) 

is a chronic cholestatic disease of the liver and bile ducts, 

with nearly 50% of patients requiring liver transplantation and 

patients having a greatly increased risk of liver and GI cancers. 

There are no proven treatments for PSC and the synthetic 

bile acid, ursodeoxycholic acid (UDCA), is toxic and is not 

recommended. The best preclinical model of PSC is the Abcb4 - 

/- 

mouse which develops liver damage due to bile acid toxicity 

- cholangitis at 2 weeks after birth, gallstone formation and 

progressive fibrosis after 8 weeks, and extensive liver damage 

leading to hepatocellular carcinoma after 12 months. Tetrahydroxylated 

bile acids (THBAs) are a novel species of bile 

acids with properties that suggest they may be significantly 

less toxic and more effective than UDCA in treating cholestasis. 

The purpose of this study was to compare the ability of 

THBAs and UDCA to prevent liver damage in Abcb4 -/- mice. 

Methods: Three isoforms of THBA were synthesized and their 

choleretic ability was measured following bile duct cannulation 

experiments in mice. Progressive liver damage in Abcb4 -/- mice 

(assessment of liver enzymes, morphology and histology) was 

determined following dietary supplementation of either THBA 

or UDCA. Results: The three isoforms of THBA studied were 

shown to be significantly more hydrophilic than UDCA and 

other known bile acids, as demonstrated by the retention time 

on a reversed-phase UPLC column. However, in mice, THBAs 

and UDCA were equally as efficient at stimulating bile flow in 

a dose-dependent fashion. Dietary supplementation of either 

THBA or UDCA in Abcb4 -/- mice showed that THBA-fed mice 

had greatly reduced gallstone formation compared to UDCAfed 

mice. Assessment of liver chemistry and histology showed 

that THBA-fed Abcb4 -/- mice had less cholestatic pressure, cholangiocyte 

damage, and fibrosis than UDCA-fed and non-fed 

controls. Conclusion: THBAs are novel bile acids that function 

as efficient choleretic agents and are more hydrophilic (and 

presumably less toxic) than UDCA. Dietary supplementation 

studies in Abcb4 -/- mice showed that THBAs can prevent progressive 

liver damage under conditions where UDCA is ineffective. 

Thus, THBAs have the potential to be used in treating 

cholestatic and other liver diseases where a therapeutic agent 

of high choleretic efficiency and low toxicity may be advantageous. 

Disclosures: Victor Ling, Renxue Wang, Jonathan Sheps 

- Patents Held/Filed: Polyhydroxylated Bile Acids for the Treatment 

of Biliary Disorders. 

Disclosures: 

Renxue Wang - Board Membership: Qing Bile Therapeutics Inc 

Jonathan Sheps - Advisory Committees or Review Panels: Qing Bile Therapeutics 

Inc.; Grant/Research Support: Qing Bile Therapeutics Inc.; Patent Held/Filed: 

Qing Bile Therapeutics Inc.; Stock Shareholder: Qing Bile Therapeutics Inc. 

Lin Liu - Stock Shareholder: Qing Bile Therapeutic Inc. 

Victor Ling - Management Position: Qing Bile Therapeutics Inc 

825 

Role of Cellular Inhibitor of Apoptosis Proteins (cIAP) in 

Primary Sclerosing Cholangitis (PSC). 

Maria Eugenia Guicciardi, Anuradha Krishnan, Steven Bronk, 

Petra Hirsova, Gregory J. Gores; Mayo Clinic, Rochester, MN 

Primary sclerosing cholangitis (PSC) is a progressive cholestatic 

liver disease of unknown etiopathogenesis. Cellular inhibitor 

of apoptosis proteins (cIAP-1 and cIAP-2) are regulators 

of death receptor signaling, and death receptor signaling 

has been implicated in the pathogenesis of PSC. We have 

recently observed that cIAP protein levels are reduced in the 

interlobular bile ducts of human PSC livers. The AIM of this 

study was to investigate whether downregulation of cIAPs is 

linked to generation of a PSC-like phenotype. METHODS: Normal 

human cholangiocyte cell lines (NHC, H69), Receptor-Interacting 

Protein 1 (RIP1)- and NF-κB2-knock-down H69 cells, 

were treated with the IAP antagonist TL-32711 (1 mM). The 

resulting samples were analyzed by apoptosis assays, immunoblot 

and qPCR. In order to verify whether inhibition of cIAPs 

in the bile ducts is sufficient to produce a PSC-like phenotype, 

IAP antagonist BV6 (100 ml/mouse) was directly instilled into 

the biliary tree. Livers and serum was analyzed by standard 

methods. Cholangiograms were obtained using a biliary cast 

and imaging by microCT with 3-D reconstruction. RESULTS: 

TL-32711 treatment induced apoptosis in 12-15% of cholangiocytes 

after 24 hr. As previously reported, the transcription 

factor NF-κB was activated following TL-32711 treatment via 

the non-canonical, RIP1-independent pathway. Notably, IAP 

antagonism led to proinflammatory cytokine (IL-8, IL-6, IL-1β) 

production in cholangiocytes, which was significantly reduced 

in cells deficient in NF-κB2 (a mediator of the non-canonical 

pathway), but not RIP1, suggesting it was largely mediated by 

the non-canonical activation of NF-κB. Five days after biliary 

injection, PSC-like changes were observed in the BV6-treated 

mice, including a fibrous cholangiopathy of the interlobular 

bile ducts, portal inflammation, significant elevation of serum 

alkaline phosphatase, total bile acids and bilirubin, and cholangiographic 

evidence of intrahepatic biliary strictures and 

dilations, and damage of the small bile ducts. CONCLUSIONS: 

We have demonstrated that decrease in cIAP levels in cholangiocytes 

induces modest cholangiocyte death, and triggers a 

significant proinflammatory response mediated by the activation 

of NF-κB in vitro. More importantly, we have generated 

an acute murine model duplicating the fibrous cholangiopathy 

of PSC by directly injecting an IAP antagonist into the biliary 

tree, suggesting that downregulation of cIAPs in cholangiocytes 

may actively contribute to the development of PSC. These data 

suggest that new therapeutic strategies aimed to prevent the 

non-canonical activation of NF-κB may be beneficial in the 

treatment of PSC. 

Disclosures: 


The following authors have nothing to disclose: Maria Eugenia Guicciardi, Anuradha 

Krishnan, Steven Bronk, Petra Hirsova 

826 

CD39/ENTPD1 is protective in a model of biliary fibrosis 

and modulates CD8+ T cell gut-homing via α4β7 integrin 

Sonja Rothweiler, Zhen-Wei Peng, Susan B. Liu, Naoki Ikenaga, 

Yury Popov, Simon C. Robson; Gastroeneterology, Beth Israel Deaconess 


BACKGROUND: Primary sclerosing cholangitis (PSC) is a 

chronic cholestatic liver disease of unknown etiology characterized 

by inflammation and fibrosis of both intra- and 

extra-hepatic bile ducts. The ectonucleotidase CD39 (ENTPD1) 

regulates purinergic signaling by hydrolizing proinflammatory 

extracellular nucleotides, such as ATP and ADP, to ultimately 

generate immunosuppressive adenosine. There are strong associations 

between PSC and inflammatory bowel disease (IBD). 

Genetic polymorphisms of CD39 were found to be linked to 

IBD, indicating CD39 to be involved in the gut immune balance. 

We studied the impact of CD39-controlled purinergic signaling


on biliary fibrosis in the Mdr2-/- mouse model, which develops 

periportal fibrosis similar to human PSC. METHODS: Histology, 

inflammatory cell infiltration, gene expression, and collagen 

architecture and content were studied in livers from Mdr2-/- 

;CD39-/- and Mdr2-/- mice. Liver infiltrating lymphocytes were 

analyzed by flow cytometry. CD8-depleting antibody was 

administered to cholic acid challenged Mdr2-/-;CD39-/- mice. 

In vitro retinoic acid (RA)-induced gut-homing α4β7 expression 

was assessed in the presence of extracellular nucleotides or 

adenosine. RESULTS: Mdr2-/-;CD39-/- mice exhibited severe 

liver injury and significantly worse portal fibrosis when compared 

to Mdr2-/- mice. The Mdr2-/-;CD39-/- mice showed 

more pronounced elevations in serum ALT levels, increase in 

total collagen levels, as determined by hydroxyproline quantification, 

and upregulated pro-fibrogenic mRNA levels in the 

liver. Flow cytometry analysis of liver infiltrating lymphocytes 

revealed a selective increase in CD8+ T cells in CD39 deficient 

Mdr2-/- mice. Further, hepatic transcription of the b7 integrin, a 

T cell gut tropism marker, was significantly increased in Mdr2- 

/-;CD39-/- mice. Antibody-mediated CD8+ T cell depletion 

in Mdr2-/-;CD39-/- mice improved outcomes with decreased 

serum alkaline phosphatase and lower expression of liver fibrosis-related 

genes, supporting the pathogenetic role of CD8+ T 

cells in PSC. RA alone was able to induce the expression of the 

α4β7 gut-homing integrin in naïve CD8+ T cells. Interestingly, 

stimulation with ATP further enhanced the RA-mediated expression 

of α4β7, whereas adenosine demonstrated the opposing 

effect by downregulating integrin expression levels. CONCLU- 

SION: Genetic deletion of CD39 exacerbates biliary injury 

and fibrosis in Mdr2-/- mice. Disease progression occurs in a 

CD8+ T cell-dependent manner and extracellular nucleotides 

increase the expression of the gut-homing recpetor α4β7. Thus, 

the purinergic system provides possible pharmacological targets 

for the treatment of PSC. 

Disclosures: 

Yury Popov - Grant/Research Support: Gilead Sciences, Inc, Takeda 





The following authors have nothing to disclose: Sonja Rothweiler, Zhen-Wei 

Peng, Susan B. Liu, Naoki Ikenaga 

827 

Feedback regulation of autotaxin in mice: why cholestatic 

mice don’t scratch 

Ruth Bolier 1 , Dagmar Tolenaars 1 , Jacqueline Langedijk 1 , Piter J. 

Bosma 1 , Ulrich Beuers 1,2 , Ronald Oude Elferink 1 ; 1 Tytgat Institute 

for Liver and Intestinal Research, Academic Medical Center, 

Amsterdam, Netherlands; 2 Department of Gastroenterology and 

Hepatology, Academic Medical Center, Amsterdam, Netherlands 

Introduction: Serum activity of autotaxin (ATX, which produces 

lysophosphatidic acid, LPA) is increased in humans during 

pregnancy and cholestasis and correlates with itch intensity 

(Gastroenterology 2010;139:1008). In contrast, serum ATX in 

mice is only marginally increased under these conditions and, 

consistently, there is no increase in scratch behavior. Aim: To 

study the difference in regulation of ATX expression between 

humans and mice. Methods: Expression of LPA receptors 

(LPAR 1-6 

) and feedback repression of ATX expression trough 

LPA receptor signaling was studied in human and mouse fibroblasts 

by qPCR. LPA signaling was stimulated with the stable 

LPA-analogue XY17 (1uM) or via endogenous ATX production 

with lipid phosphate phosphatase 1 inhibitor XY14 (10uM), to 

prevent the rapid breakdown of LPA. LPA production by endogenous 

ATX was inhibited by ATX inhibitor HA115 (10uM) and 

LPAR 1-3 

signaling was blocked with the receptor antagonist 

Ki16425 (25uM). Involvement of G-proteins and their downstream 

pathways was investigated using the cAMP stimulator 

forskolin (25uM), protein kinase A inhibitor RP-adenosine 

(100uM), G α 

i/o-inhibitor pertussis toxin (500ng/mL), inositosol 

1,4,5-triphophate inhibitor U73122 (1uM) and protein 

kinase C inhibitor Gö6983 (10uM). In addition, the established 

induction of ATX by TNFα (10ng/mL) was compared 

in fibroblasts of the two species. Results: TNFα stimulated 

ATX expression 3- to 4-fold in fibroblasts from both species. In 

murine cells, stimulation of LPAR-signaling by XY14 and XY17 

repressed ATX mRNA expression by 70 and 82%, respectively 

(both p


bated cleaved caspase-3 enzyme activity and overexpression 

of pJNK and RIP3 was characteristic of liver explants from 

PBC patients compared with healthy controls, accompanied 

by elevated protein levels of TNF and IL-6. Casp8 Δhepa after 

BDL displayed decreased number and size of necrotic foci 

compared with Casp8 f/f mice. Significantly decreased serum 

alanine (ALT) and aspartate (AST) transaminases, TUNEL 

staining, and cleaved Caspase-3 and cytochrome C protein 

overexpression were found in Casp8 Δhepa mice 28 days after 

BDL, along with diminished compensatory proliferation (Ki-67, 

PCNA) and ductular reaction (CK-19). These results were translated 

into a decreased inflammatory profile elicited by lower 

IL-6, TNF-α protein levels and reduced infiltration of F4/80 + , 

Cd11b + and CD45 + populations. Overall, liver fibrogenesis 

(Sirius red, Collagen IA1, ASMA) was significantly ameliorated 

in Casp8 Δhepa . Mechanistically, decreased activation of 

JNK, RIP1 and RIP3 correlated with the protection shown after 

chronic BDL. Conclusions: CASP8 in hepatocytes is an essential 

orchestrator of cholestatic liver injury and its specific modulation 

might be an interesting pharmacologic target that can be 

used in the clinic as treatment for obstructive liver disease. 

Disclosures: 



The following authors have nothing to disclose: Francisco Javier Cubero, Jin 

Peng, Maximilian Hatting, Gang Zhao, Miguel Eugenio Zoubek, Ricardo 

Macías-Rodríguez, Astrid Ruiz-Margáin, Johanna Reissing, Henning W. Zimmermann, 

Nikolaus Gassler, Tom Luedde, Christian Liedtke 

829 

NHERF-1 assembles macromolecular complexes in the 

liver that are essential for the inflammatory response in 

cholestasis 

Man Li 1 , Albert Mennone 1 , Carol J. Soroka 1 , Lee R. Hagey 3 , 

Kathy M. Harry 1 , Edward J. Weinman 2 , James L. Boyer 1 ; 1 Internal 

Medicine, Yale University School of Medicine, New Haven, CT; 

2 University of Maryland School of Medicine, Baltimore, MD; 3 University 

of California, San Diego, La Jolla, CA 

The Na+/H+ exchanger regulatory factor 1 (NHERF-1/EBP50) 

is a PDZ protein that has been shown to participate in signal 

transduction by interacting with signaling molecules such as G 

protein-coupled receptors as well as regulatory proteins such 

as protein kinases. Our previous studies showed that deletion 

of NHERF-1 in mice leads to reduced hepatic expression of 

cytoskeletal ezrin-radixin-moesin (ERM) proteins and intercellular 

adhesion molecule-1 (ICAM-1), a molecule that plays a key 

role in neutrophil-mediated liver injury in mice after bile duct 

ligation (BDL). NHERF-1-/- BDL mice have significantly lower 

scores of hepatic necrosis, serum ALT, as well as reduced neutrophil 

accumulation in the liver compared with the wild-type 

(WT) BDL group. Aims: To further investigate the mechanisms 

of protection of liver injury induced by BDL in NHERF-1-/- mice. 

METHODS: Co-Immunoprecipitation was performed to identify 

protein complexes that are formed with NHERF-1 in the liver 

of sham and 7 day BDL WT mice. Immunoblotting was utilized 

to compare protein expression in sham and BDL WT and 

NHERF-1-/- mouse liver. Hepatic bile acids were analyzed 

by mass spectrometry as well. RESULTS: NHERF-1 co-immunoprecipitated 

with ICAM-1 in mouse liver, assembling ERM 

proteins, ICAM-1 and F-actin into a macromolecule complex 

that is increased in mouse liver and participates in neutrophil 

infiltration after BDL. In the liver, NHERF-1 also interacts with 

PKCζ, an atypical PKC that has been shown to phosphorylate 

and activate NF-κB p65. Compared with the WT control, 

NHERF-1-/- mice have reduced expression of hepatic PKCζ. 

Expression of total NF-κB p65 as well as phosphorylated NF-κB 

p65 was significantly reduced in the liver of NHERF-1-/- BDL 

mice compared with WT BDL mice. While total bile acid concentrations 

in the serum and liver of sham and BDL NHERF-1- 

/- mice were not significantly different from the WT controls, 

hepatic tetrahydroxylated bile acids and Cyp3a11 mRNA levels 

were significantly higher in NHERF-1-/- BDL mice, indicating 

a more profound adaptive response of the hepatocytes to 

BDL in these mice. CONCLUSIONS: NHERF-1 participates in the 

inflammatory and adaptive responses that is associated with 

BDL induced liver injury. Deletion of NHERF-1 in mice leads 

to disruption of the formation of ICAM-1-ERM-NHERF-1 and 

PKCζ/NHERF-1 complexes, causing reduction of hepatic ERM 

proteins, ICAM-1 and PKCζ, molecules that are essential for 

the inflammatory response in cholestasis. Further study of the 

role of NHERF-1 in the inflammatory response in cholestasis 

and other forms of liver injury should lead to discovery of new 

therapeutic targets in hepatic inflammatory diseases. 

Disclosures: 


abbvie 

The following authors have nothing to disclose: Man Li, Albert Mennone, Carol J. 

Soroka, Lee R. Hagey, Kathy M. Harry, Edward J. Weinman 

830 

Knockdown of α7-Nicotinic Receptor Inhibits Biliary 

Proliferation and Hepatic Fibrosis during Extrahepatic 

Cholestasis 

Allyson Martinez 1 , April O’Brien 1 , Laurent Ehrlich 1 , David E. Dostal 

1 , Shannon S. Glaser 1,2 ; 1 Department of Medicine, Texas A&M 

Health Science Center and Central Texas Veterans Health Care 

System, Temple, TX; 2 Scott & White Digestive Disease Research 

Center, Temple, TX 

In cholestatic liver diseases, cholangiocytes, through the products 

of their cellular activation, are implicated as the key link 

between bile duct injury and the subepithelial fibrosis that 

characterizes chronic hepatobiliary injury. We have previously 

demonstrated that activation of α7-nicotine receptor (nAChR) 

with nicotine stimulates cholangiocyte proliferation that was 

associated with increased profibrotic gene expression by cholangiocytes 

and deposition of collagen in portal areas in normal 

wild-type mice. We have also demonstrated that mechanical 

stress, which occurs due to increased biliary pressure during 

extrahepatic cholestasis, stimulates cholangiocyte proliferation. 

The role of the α7-nAChR during extrahepatic cholestasis or 

mechanical stress has not been thoroughly explored. Thus, the 

AIM of our study was to determine the role of α7-nAChR in the 

regulation of biliary proliferation and hepatic fibrosis during 

extrahepatic cholestasis induced by bile duct ligation (BDL). 

Methods: Studies were performed in normal and BDL (1 week) 

wild-type (WT) and α7-nAChR knockout (KO) mice. Intrahepatic 

bile duct mass (IBDM) and biliary proliferation was evaluated 

by immunohistochemistry for CK-19 and PCNA in liver 

sections and by immunoblots in isolated cholangiocytes. Liver 

fibrosis was evaluated by Sirius red staining in liver sections 

and by qPCR for the profibrotic markers [collagen 1 (COL1A1), 

fibronectin (FN-1), and αSMA] in isolated cholangiocytes and 

total liver samples. In vitro, mouse primary cholangiocytes were 

plated on BioFlex culture plates and static equiaxial strain was 

applied to the cells for 24 hrs in the presence and absence of 

shRNA to knockdown α7-nAChR and the α7-nAChR antagonist 

methyllycaconitine (1.4 nM) . Proliferation was evaluated by 

immunoblots for PCNA and the expression of COL1A1, FN-1 

and αSMA by qPCR. Results: In vivo, there was a significant 

reduction in biliary proliferation (PCNA) and IBDM (CK-19) 

in BDL α7-nAChR KO mice compared to BDL WT. In addi-


tion to a reduction in proliferation, there was also a reduction 

in hepatic fibrosis by Sirius red staining and a significant 

reduction in COL1A1, FN-1 and αSMA gene expression. In 

vitro, knockdown or inhibition of α7-nAChR in mouse cholangiocytes 

prevented mechanical stress-induced proliferation. 

Knockdown or inhibition of α7-nAChR also inhibited mechanical 

stress-induced COL1A1, FN-1 and αSMA gene expression. 

Conclusion: The knockout of α7-nAChR inhibits BDL induced 

proliferation and hepatic fibrosis. Modulation of the α7-nAChR 

axis during extrahepatic cholestasis may represent a novel therapeutic 

approach for cholestatic liver diseases. 

Disclosures: 

The following authors have nothing to disclose: Allyson Martinez, April O’Brien, 

Laurent Ehrlich, David E. Dostal, Shannon S. Glaser 

831 

Keratin 23 represents a novel liver injury marker reflecting 

the severity of ductular reaction 

Nurdan Guldiken 1 , Gokce Kobazi Ensari 1 , Pooja Lahiri 2 , Christian 

Liedtke 1 , Henning W. Zimmermann 1 , Christian Trautwein 1 , 

Marianne Ziol 3 , Pavel Strnad 1 ; 1 Internal Medicine III, Aachen, 

Germany; 2 Institut für Pathologie, Graz, Austria; 3 Pathology 

Department, Paris, France 

Background: Keratins (K) are the intermediate filaments of epithelial 

cells and constitute established diagnostic tools. In the 

liver, K7/K19 expression is restricted to hepatic progenitor 

cells (HPCs), intermediate (i.e. not fully differentiated) hepatocytes 

and biliary epithelial cells (BECs). Consequently, K7/ 

K19 represent a widely used marker of the regenerative liver 

response termed ductular reaction (DR) that consists of activated 

BECs and HPCs. Methods: Given that K23 is a largely 

unknown keratin family member, we analysed its expression 

and localization in selected human liver disorders (8 controls, 

15 patients with simple alcoholic steatosis [ALD], 9 with 

chronic hepatitis B/ 13 with chronic hepatitis C (HCV), 12 with 

non-alcoholic steatohepatitis; 7 with acute liver failure [ALF]; 

7 with end-stage primary biliary cirrhosis [PBC]) and mouse 

liver injury models using custom-made antibodies. Results: In 

untreated mice, K23 was found in biliary epithelia but not 

hepatocytes. It was (together with K7/K19) markedly up-regulated 

in three different DR/cholestatic injury models, i.e. four 

months old MDR2 knockouts, animals treated with 3,5-diethoxycarbonyl-1,4-dihydrocollidine 

for four weeks or subjected to 

bile duct ligation for five days. No changes in K23 levels were 

seen in hepatocellular injury models such as partial hepatectomy 

or carbon tetrachloride-induced fibrogenesis. K23 levels 

correlated with the DR marker Fn14 and immunofluorescence 

staining showed a distinct but not perfect co-localization with 

K7/K19. In humans, K23 levels were moderately up-regulated 

in active HCV (~3 times) and ALD (~10 times). K23 expression 

was higher in patients with more prominent inflammation/fibrosis. 

A dramatic increase (>200 times) was observed in patients 

with ALF and end-stage PBC. In ALF/PBC, K7/K23 positive, 

K19 negative cells corresponding to intermediate hepatocytes 

were frequently found. In conclusion, K23 represents a novel 

DR marker and its levels correlate with the severity of the underlying 

human disease. 

Disclosures: 



The following authors have nothing to disclose: Nurdan Guldiken, Gokce Kobazi 

Ensari, Pooja Lahiri, Christian Liedtke, Henning W. Zimmermann, Marianne Ziol, 

Pavel Strnad 

832 

RIP3-dependent signalling contributes to necroinflammation 

in cholestatic liver injury 

Marta B. Afonso 2 , Pedro M. Rodrigues 2 , André L. Simão 2 , Helena 

Cortez-Pinto 3,4 , Dimitry Ofengeim 5 , Joana D. Amaral 1 , Rui E. 

Castro 1 , Junying Yuan 5 , Cecília M. Rodrigues 1 ; 1 iMed.ULisboa & 

Dep. of Biochemistry and Human Biology, Faculty of Pharmacy, 

University of Lisbon, Lisboa, Portugal; 2 iMed.ULisboa, Faculty of 

Pharmacy, University of Lisbon, Lisbon, Portugal; 3 Gastrenterology, 

Hospital de Santa Maria, Lisbon, Portugal; 4 Faculty of Medicine, 

University of Lisbon, Lisbon, Portugal; 5 Department of Cell Biology, 

Harvard Medical School, Boston, MA 

Cholestasis is a common pathological condition characterized 

by retention of bile acids in liver and serum, with concomitant 

liver inflammation and hepatocyte damage. Recent evidence 

suggests that regulated necrosis or necroptosis, which depends 

on the kinase activity of receptor interacting protein 3 (RIP3), 

may contribute to the pathogenesis of inflammation-driven 

liver diseases. Here we aimed to evaluate the role of necroptosis 

after common bile duct ligation (BDL) in mice, a classic 

experimental model for acute cholestasis and secondary 

biliary fibrosis, and in patients with primary biliary cirrhosis 

(PBC), a cholestatic chronic liver disease. RIP3 and its target 

mixed lineage kinase domain-like protein (MLKL), were evaluated 

by immunohistochemistry in liver specimens of patients 

with PBC and healthy volunteers. BDL or sham surgery was 

performed in C57BL/6 wild-type (WT) or RIP3-deficient (RIP3 - 

/- ) mice. Serum and livers were collected 3 and 14 days after 

BDL. Liver histology and serum liver enzymes were evaluated. 

Liver RIP3, proinflammatory and fibrotic markers were determined 

by qRT-PCR. Total and soluble/insoluble liver proteins 

were analysed by Western blot. RIP3 kinase activity was determined 

in vitro. Finally, the functional crosstalk between RIP3 

and antioxidant response was investigated. RIP3 and phosphorylated 

MLKL protein levels were upregulated in the liver of 

PBC patients (p < 0.05). In WT mouse livers, BDL resulted in 

progressive bile duct hyperplasia, multifocal necrosis, fibrosis 

and inflammatory cell infiltration. Concomitantly, necroptosis 

was activated as assessed by: 1) increased RIP3 mRNA and 

protein expression; 2) augmented RIP3 kinase activity, and 3) 

sequestration of RIP3 and MLKL in the insoluble protein fraction 

of the liver (at least p < 0.05). Remarkably, histological analysis 

revealed that absence of RIP3 significantly decreased liver 

necrosis and inflammation induced by BDL at both time-points 

(p < 0.05). Accordingly, at day 3, BDL RIP3 -/- mice showed 

lower circulating levels of hepatic enzymes and decreased 

inflammatory and fibrogenic liver gene expression, together 

with enhanced antioxidant response (at least, p < 0.05). Protection 

was no longer evident at 14 days after BDL in RIP3 -/- . 

In conclusion, RIP3-dependent signaling is triggered in human 

PBC and mediates hepatic necroinflammation in BDL-induced 

acute cholestasis. Targeting necroptosis may be a promising 

therapeutic approach to treat cholestatic liver injury, although 

complementary approaches may be required to control progression 

to fibrosis. 

Disclosures: 

Helena Cortez-Pinto - Advisory Committees or Review Panels: Norgine, Lundbeck; 

Speaking and Teaching: Janssen, Gilead Janssen 

The following authors have nothing to disclose: Marta B. Afonso, Pedro M. 

Rodrigues, André L. Simão, Dimitry Ofengeim, Joana D. Amaral, Rui E. Castro, 

Junying Yuan, Cecília M. Rodrigues


833 

Gut microbiota contributes to disease in a spontaneous 

mouse model of chronic cholangitis 

Elisabeth Schrumpf 1,2 , Martin Kummen 1,2 , Kristian Holm 1,2 , Tom 

H. Karlsen 1,2 , Johannes R. Hov 1,2 , Richard S. Blumberg 3 , Espen 

Melum 1,2 ; 1 Norwegian PSC Research Center, Division of Cancer, 

Surgery and Transplantation, Oslo University Hospital, Rikshospitalet, 

Oslo, Norway; 2 K.G. Jebsen Inflammation Research Centre, 

Research Institute of Internal Medicine, Oslo University Hospital, 

Rikshospitalet, Oslo, Norway; 3 Division of Gastroenterology, 

Hepatology and Endoscopy, Department of Medicine, Brigham 

and Women’s Hospital, Harvard Medical School, Boston, MA 

Purpose: NOD.c3c4 mice are on a NOD background and 

spontaneously develop biliary inflammation both in extra- and 

intra-hepatic bile ducts. Gut microbes and lymphocytes activated 

in the intestines are hypothesized to play a role in the 

human biliary disease primary sclerosing cholangitis. We 

aimed to clarify the role of the gut microbiota in NOD.c3c4 

mice. Methods: We sampled cecal content and mucosa from 

10 week old NOD.c3c4 (n=5) and NOD mice (n=5) housed 

in a minimal disease unit (MDU) facility. The NOD.c3c4 and 

NOD strains were rederived into a new MDU facility, and 

after three generations the experiment was repeated. DNA 

was extracted and the V4 region of the 16S rRNA gene was 

amplified and sequenced on the Illumina MiSeq. Data were 

analysed with Quantitative Insights Into Microbial Ecology 

(QIIME). NOD.c3c4 mice were also rederived into a germ free 

(GF) facility. Conventionally housed (n=9) and GF NOD.c3c4 

mice (n=9) were harvested at 9 weeks of age, with sampling of 

the liver and blood. Results: The gut microbial profiles of mice 

with and without biliary disease were different both before and 

after rederivation (beta-diversity, unweighted UniFrac-distance, 

anosim: p


835 

Knockdown of the melatonin receptor, MT1, reduces 

biliary hyperplasia and liver fibrosis in cholestatic bile 

duct ligated (BDL) mice 

Nan Wu 2 , Fanyin Meng 1 , Ying Wan 3 , Julie Venter 2 , Holly A. Standeford 

4 , Heather L. Francis 1 , Lindsey Kennedy 4 , Yuyan Han 1,2 , 

Kelly McDaniel 3 , Antonio Franchitto 5 , Paolo Onori 5 , Sharon 

DeMorrow 1 , Tami Annable 3 , Eugenio Gaudio 5 , Shannon S. Glaser 

1 , Gianfranco Alpini 1 ; 1 Research, S&W DDRC and Medicine, 

Central Texas Veterans Health Care System, Scott & White and 

Texas A & M Health Science Center College of Medicine and 

Scott White, Temple, TX; 2 Medicine, Texas A&M University HSC, 

Temple, TX; 3 Operational Funds, BaylorScott&White, Temple, TX; 

4 Research, Central Texas Veterans Health Care System, Temple, 

TX; 5 Department of Anatomical, Histological, Forensic Medicine 

and Orthopedics Sciences, University La Sapienza, Rome, Italy 

Cholestatic liver diseases are characterized by increased biliary 

proliferation and intrahepatic bile duct mass (IBDM), liver 

damage and fibrosis. Melatonin, which is synthesized in the 

pineal gland as well as cholangiocytes, exerts its effects by 

interaction with MT1 and MT2 receptors. Knockdown of MT2 

increases biliary proliferation and liver fibrosis in cholestatic 

mice. Also, melatonin inhibits biliary hyperplasia by interacting 

with MT1 receptors. However, since these studies were 

performed only in vitro, we evaluated the role of MT1 in regulating 

biliary proliferation and liver fibrosis in vivo in MT1 

knockout (KO) mice, and wild-type (WT) mice in which the 

hepatic expression of MT1 was reduced by Vivo-Morpholinos 

for MT1. Methods: The studies were performed in normal (WT), 

MT1 Morpholino-treated or MT1 KO mice with or without BDL 

surgery. We evaluated the expression of MT1 in liver sections 

by immunohistochemistry (IHC) and immunoblots in cholangiocytes. 

Intrahepatic biliary ductal mass (IBDM) and biliary proliferation 

was evaluated by IHC for CK-19 and PCNA in liver 

sections, and qPCR for PCNA in cholangiocytes. Liver injury 

was determined by H&E staining in liver sections and serum 

levels of transaminases and bilirubin. Fibrosis was evaluated 

by Sirius red staining in liver sections and qPCR for TGFbeta1, 

alpha-SMA and fibronectin in cholangiocytes and total liver. In 

vitro, shRNA-MT1 transfected murine cholangiocytes (MCCs) 

and controls were used to measure proliferation and TGFbeta1, 

alpha-SMA and fibronectin (FN-1) mRNA expression by qPCR. 

Results: Normal cholangiocytes express low levels of MT1, 

which markedly increased after BDL. We demonstrated that in 

MT1 KO BDL and MT1 Morpholino-treated BDL mice, along 

with reduced MT1 expression, there was decreased biliary 

proliferation and IBDM. In MT1 KO BDL and MT1 Morpholino-treated 

BDL mice, there was decreased lobular damage, 

serum levels of transaminases and bilirubin, and liver fibrosis in 

liver sections and reduced expression of TGFbeta1, alpha-SMA 

and FN-1 compared to BDL WT mice. In shRNA-MT1 transfected 

MCCs, there was decreased proliferation and reduced 

expression of TGFb1, alpha-SMA and FN-1. The findings indicate 

that shifting the melatonin axis to signal via MT2 during 

the knockdown of MT1 expression decreases biliary proliferation 

and liver fibrosis. Conclusion: Our findings indicate that 

the balance of MT1/MT2 receptor expression for the melatonin 

signaling axis may play an important role in the regulation of 

biliary proliferation and hepatic fibrosis. Inhibition of MT1 may 

be a key approach for ameliorating biliary hyperplasia and 

liver fibrosis in cholestatic liver diseases. 

Disclosures: 

The following authors have nothing to disclose: Nan Wu, Fanyin Meng, Ying 

Wan, Julie Venter, Holly A. Standeford, Heather L. Francis, Lindsey Kennedy, 

Yuyan Han, Kelly McDaniel, Antonio Franchitto, Paolo Onori, Sharon DeMorrow, 

Tami Annable, Eugenio Gaudio, Shannon S. Glaser, Gianfranco Alpini 

836 

Longer Term Efficacy of Simultaneous Liver Transplantation 

plus Sleeve Gastrectomy versus non-invasive 

Weight Loss followed by Liver Transplantation 

Daniel Zamora-Valdes 1 , Kymberly D. Watt 2 , John J. Poterucha 2 , 

Charles B. Rosen 1 , Todd A. Kellogg 3 , Sara R. Di Cecco 2 , Nicki 

M. Francisco Ziller 2 , Julie Heimbach 1 ; 1 Transplant Surgery, Mayo 

Clinic, Rochester, MN; 2 Liver Transplant, Mayo Clinic, Rochester, 

MN; 3 General Surgery, Mayo Clinic, Rochester, MN 

Obesity is increasingly common before and after liver transplantation 

(LT), yet optimal management remains unclear. Our 

AIM was to analyze the long-term effectiveness of a multi-disciplinary 

protocol for obese patients requiring LT, including 

a non-invasive pre-transplant weight loss program, and combined 

LT plus sleeve gastrectomy (SG) for obese patients who 

failed to lose weight prior to LT. METHODS: since 2006, all 

patients referred for LT with a BMI >35 were enrolled. There 

are 46 patients who achieved weight loss and underwent LT 

alone, and 17 who underwent LT combined with SG. Outcomes 

for patients with > 2 years since LT alone or LT+SG 

were analyzed using Student T-test for continuos variables and 

c 2 for categorical variables. RESULTS: In those who received 

LT alone, 34/46 were successful on achieving >10% loss in 

total body weight (TBW) prior to LT. Of those with > 2 years 

since LT, weight gain to BMI>35 was seen in 21/34 (61.7%) 

at 2 years (Figure 1), though 10/34 (30%) who maintained 

>10% loss in TBW at 2 years (p=0.005; Figure 1). There were 

8/34 (23.5%) with diabetes post LT and 10/34 (18.9%) with 

evidence of steatosis on year 2 ultrasound. In patients undergoing 

the LT+SG, all patients achieved significant weight loss 

(mean BMI at listing 49.8±1.5 vs. 2 year post LT 29.3±6.4; 

p=0.001). No LT+SG patient developed post-LT DM or steatosis 

at 2 years post LT. Conclusion: Non-invasive pre-transplant 

weight loss was achieved by a majority, though weight gain 

post LT was common. Combined LT plus SG resulted in effective 

weight loss and was associated with fewer post-LT metabolic 

complications at 2 years post LT. 

Figure 1: post transplant BMI in morbidly obese patients who 

underwent medical management (n=34) or sleeve gastrectomy 

(n=5) combined with liver transplantation. 

Disclosures: 

The following authors have nothing to disclose: Daniel Zamora-Valdes, Kymberly 

D. Watt, John J. Poterucha, Charles B. Rosen, Todd A. Kellogg, Sara R. Di 

Cecco, Nicki M. Francisco Ziller, Julie Heimbach


837 

Preliminary Data Analysis from the Improving DCD Outcomes 

in Liver Transplantation (IDOL) Consortium 

David S. Goldberg 1 , Seth J. Karp 3 , Julie Heimbach 5 , James F. 

Markmann 6 , Richard Gilroy 4 , Roberto Hernandez-Alejandro 7 , 

Peter L. Abt 2 ; 1 Division of Gastroenterology, Hospital of the University 

of Pennsylvania, Philadelphia, PA; 2 Department of Surgery, 

University of Pennsylvania, Philadelphia, PA; 3 Department 

of Surgery, Vanderbilt University, Nashville, TN; 4 Department of 

Medicine, University of Kansas, Kansas City, KS; 5 Department of 

Surgery, Mayo Clinic, Rochest, MN; 6 Department of Surgery, Massachusetts 

General Hospital, Boston, MA; 7 Department of Surgery, 

London Health Sciences Center, London, ON, Canada 

Introduction: Increased utilization of donation after cardiac 

death (DCD) liver allografts could help meet the demand for 

transplantable livers. Current understanding of post-liver transplant 

(LT) outcomes in DCD recipients is limited to single-center 

reports, or national analyses of UNOS data that do not capture 

detailed biliary complication data. We created the IDOL 

Consortium to collect multi-center patient-level data to: 1) overcome 

limitations of prior DCD analyses; 2) validate previous 

single-center studies; and 3) identify novel factors associated 

with DCD outcomes. Methods: The consortium includes 10 US 

centers and 2 centers in Canada. Detailed patient-level data 

were entered into a Redcap database. Donor and recipient 

age, and donor warm ischemic time (WIT; extubation to crossclamp) 

were evaluated as primary exposures, with primary 

outcomes of biliary complications (BC; excluding bile leak) 

and graft failure within 6 months post-DCD transplantation. 

Results: To date, 171 DCD recipients have been entered into 

the IDOL Consortium database. The median recipient age 

was 57 years (IQR: 50-61), 117 (68.4%) were male, and the 

median lab MELD was 19 (IQR: 13-24). The median donor 

WIT was 25 minutes (IQR: 20-30). 41 (24.0%) patients had 

a BC within the first 6 months post-LT: 18 (43.9%) with diffuse 

intrahepatic strictures (ischemic cholangiopathy), 14 (34.1%) 

anastomotic strictures, 7 (17.1%) with ischemic intrahepatic 

strictures and an anastomotic stricture, and 2 (4.9%) had ischemic 

extra-hepatic non-anastomotic strictures. There were 21 

(12.3%) graft failures, and 18 (11.2%) deaths within the first 

6 months post-LT. Of these 21 graft failures, 4 (19.0%) had a 

history of a BC. Donor age >40 years of age was numerically, 

but not statistically, associated with increased risk of BCs (OR: 

1.86, 95% CI: 0.91-3.77, p=0.09), while a donor WIT>30 

minutes was associated with significantly increased risk of BCs 

(OR: 2.41, 95% CIL 1.06-5.47, p=0.04). Increase recipient 

age was associated with increased risk of graft failure (OR 

for every 5-year increase: 1.49, 95% CI: 1.06-2.10), while 

donor WIT>30 minutes was numerically, but not statistically 

associated with graft failure (OR: 2.48, 95% CI: 0.93-6.64, 

p=0.07). Laboratory MELD score was not associated with BC 

or graft failure. Discussion: Preliminary data from the IDOL 

Consortium provide estimates of BCs among a diverse cohort 

of DCD recipients, and identify potential donor and recipient 

factors associated with BCs and graft failure. Completion of 

the IDOL Consortium data collection is anticipated in the next 3 

months, which will allow for sufficient sample sizes to estimate 

all key exposures and outcomes. 

Disclosures: 


The following authors have nothing to disclose: David S. Goldberg, Seth J. Karp, 

Julie Heimbach, James F. Markmann, Roberto Hernandez-Alejandro, Peter L. Abt 

838 

A shift to the left: A comparison of the A2ALL experience 

using left lobe vs. right lobe grafts 

Benjamin Samstein 1 , Abigail Smith 9 , Talia B. Baker 8 , Chris 

Freise 12 , Jean C. Emond 2 , Brenda W. Gillespie 9 , David Grant 4 , 

Elizabeth A. Pomfret 11 , Adrian Cotterell 3 , Trevor L. Nydam 6 , 

Kim M. Olthoff 7 , Abhinav Humar 10 , Robert M. Merion 5 ; 1 Department 

of Surgery, Weill Cornell Medical Center, New York, NY; 

2 Columbia University, New York, NY; 3 Virginia Commonwealth 

University, Richmond, VA; 4 University of Toronto, Toronto, ON, 

Canada; 5 Arbor Research Collaborative for Health and University 

of Michigan Department of Surgery, Ann Arbor, MI; 6 Department 

of Surgery, University of Colorado School of Medicine, Aurora, 

CO; 7 University of Pennsylvania, Philadelphia, PA; 8 Comprehensive 

Transplant Center, Northwestern University Feinberg School 

of Medicine, Chicago, IL; 9 University of Michigan, Ann Arbor, MI; 

10 University of Pittsburgh, Pittsburgh, PA; 11 Lahey Hospital and 

Medical Center, Burlington, ME; 12 Transplant Division, University 

of California, San Francisco, San Francisco, CA 

Exploration of the use of left lobe grafts was a principal aim 

of A2ALL. Potential living donor liver transplant (LDLT) recipients 

and their donors were enrolled at 12 North American 

centers between 2004 and 2014 with transplants occurring 

between 1998 and 2014. Of the 1141 donors and 1136 

recipients enrolled 99 were left lobe donors and 88 were left 

lobe recipients, respectively. Preoperative characteristics, intraoperative 

and postoperative data including patient and graft 

survival were compared by lobe using t-tests, chi-square tests, 

Wilcoxon tests, and Cox models. Characteristics of graft type 

and outcomes were analyzed over time and between centers. 

There was a marked center effect in the use of left lobes, with 

two centers accounting for 73% and 71% of left lobe recipients 

and donors, respectively. Right and left lobe donors did not 

differ significantly with respect to clinical characteristics. Left 

lobe donor surgeries were more likely to be laparoscopic (55% 

vs. 31%, p=0.003). Postoperatively, left lobe donors had lower 

bilirubin (0.9±0.6 vs. 1.6±1.3, p


fact that all obese patients remained obese post-OLT without 

a significant difference in BMI pre- and post-OLT regardless 

of their pre-OLT category. In conclusion, our findings point to 

obesity being a major risk of NAFLD recurrence in liver graft 

post-OLT. 

Disclosures: 

Roberto J. Firpi - Advisory Committees or Review Panels: Quest Diagnostics, 

Gilead; Grant/Research Support: Bayer, Vertex, BMS, Janssen, Gilead, Merck 

The following authors have nothing to disclose: Angela Dolganiuc, Vikas Khullar, 

Virginia C. Clark 

Disclosures: 

The following authors have nothing to disclose: Benjamin Samstein, Abigail 

Smith, Talia B. Baker, Chris Freise, Jean C. Emond, Brenda W. Gillespie, David 

Grant, Elizabeth A. Pomfret, Adrian Cotterell, Trevor L. Nydam, Kim M. Olthoff, 

Abhinav Humar, Robert M. Merion 

839 

Obesity is predictive of NAFLD recurrence in the transplanted 

liver graft. 

Angela Dolganiuc, Vikas Khullar, Virginia C. Clark, Roberto J. 

Firpi; Medicine/GI, University of Florida, Gainesville, FL 

Non-Alcoholic Fatty Liver Disease (NAFLD) is an emerging 

forerunning cause of liver transplant (OLT) in USA and worldwide. 

We have previously reported that cryptogenic cirrhosis 

category masks three quarters of NASH patients getting OLT 

prior to 2013. We aimed to identify clinical and routine biochemical 

features predictive of NAFLD recurrence post-OLT. 

Our cohort includes patients from the University of Florida 

Liver Transplant database from 1990-2013. 174 out of 1,646 

patients were listed for cryptogenic cirrhosis (CC, 156 patients) 

or NASH cirrhosis (18 patient, pre-OLT known) as cause of 

OLT. In the CC group, 116 patients had electronic records; 

40 were excluded from analysis based on explant histology 

non-consistent with CC or NAFLD. The remaining 76 patients 

were further distributed, based on explant liver histology, into 

True CC (24 patients which did not have inflammation or steatosis, 

used as control), and NASH (consisting of a subgroup 

of n=24 active NASH with steatosis and inflammation, and 

n=28 late-stage NASH with inflammation but no steatosis) categories. 

There were no differences between the NASH cohort, 

CC group-derived NASH and true-CC cohort in terms of age, 

gender, or ethnicity or the frequency and characteristics of 

metabolic syndrome. NAFLD (all histological stages combined) 

recurrence was observed in 42% of the per-OLT known NAFLD 

cohort (follow-up interval 3.2±2.8 years), 32% in CC group-derived 

NASH with steatosis (follow-up interval 3.6±2.4 years) 

and 16% of CC group-derived NASH without steatosis (follow-up 

interval 1.2±1.1 years). In contrast, none of the true 

CC patients developed NAFLD post-OLT within the follow-up 

interval 3.2±2.8 years. Analysis of metabolic syndrome (blood 

pressure, lipid profile and fasting glucose) profile revealed 

that pre-transplant lipid profile (HDL, LDL, Cholesterol, Tg) was 

abnormally low in all groups pre-OLT, likely reflective of pre- 

OLT malnutrition, and was not predictive of NAFLD recurrence. 

Similarly, systolic blood pressure and presence of DM pre-OLT 

were not predictive of NAFLD recurrence in the liver graft. 

There were significantly more obese patients in the pre-OLT 

known NASH cohort (98%) and in CC group-derived NASH 

with (86%) or without (83%) steatosis compared to true-CC 

(69%) cohort. More importantly, all patients who had NAFLD 

recurrence in their liver graft were obese pre-OLT, despite the 

840 

3D Printed Liver Models for Precise Resection of Complex 

Hepatic Tumors 

Maggie H. Samaan, Bijan Eghtesad, Cristiano Quintini, Juliana 

Kissiedu, Ibrahim A. Hanouneh, Ryan Klatte, Charles M. Miller, 

Nizar N. Zein; Gastroenterology and Hepatology, Digestive Disease 

Institute, C.C.F, Cleveland, OH 

We recently described a novel method to obtain a patient specific 

3D printed liver model s and demonstrated applicability 

in living donor liver transplantation [1]. Hepatic resection for 

the treatment of liver tumors is an accepted therapy although 

associated with a number of limitations due to incomplete characterization 

of anatomy and proximity of tumor to vascular and 

biliary structures. 3D printed liver models offer better representation 

of spatial relationships and improve surgical outcome. 

The purpose of this study is to compare 2D images (CT, MRI) to 

3D printed liver models for preoperative surgical planning and 

intraoperative guidance in complex hepatic tumor resection. 

Methods: Prospective study of 6 consecutive patients with complex 

liver tumors (located centrally, infiltrating/abutting main 

HVs or requiring non-anatomical resections/intrahepatic vasculature 

reconstruction). Median lesion size was 7.1cm (range 

4.7–11cm). Preoperative evaluation included standard CT & 

MRI. A patient-specific 3D printed liver was developed for each 

case. For assessment of preoperative planning, a surgical plan 

was developed for each case using standard imaging (Plan 

A) and was revised after the surgical team was provided with 

the 3D printed liver specific to the case (plan B). 3D models 

were available during the operation as a reference to guide 

resection as deemed necessary by the surgical team in addition 

to standard intraoperative ultrasound. 3D printing process 

included 3D image reconstruction, digital preparation, 3D 

printing and post-printing processing. RESULTS: In 3 of the 6 

cases, the pre-operative plan was modified after review of anatomical 

spatial relationship of tumor to nearby structures in the 

3D model compared to initial plan based on standard imaging 

alone. Changes included resection modification, extension and 

intrahepatic vascular reconstruction. Intraopertively, surgeons 

reported greater confidence with use of 3D model for identification 

of intra and extrahepatic structure, segmentation and 

tumor specific extent. Surgeons agreed that 3D model offered 

a realistic representation that allowed interactive manipulation 

simulating intraoperative mobilization. 3D printed liver models 

offer a realistic patient-specific representation that may improve 

surgical planning and intraoperative process in patients with 

complex hepatic tumors. [1] Zein NN, et al. Liver Transpl 

2013;19:1304-10. Picture 1: to be submitted upon request 

(couldn’t be submitted for it exceeded maximum allowed characters, 

needs lower resolution) 

Disclosures: 

The following authors have nothing to disclose: Maggie H. Samaan, Bijan Eghtesad, 

Cristiano Quintini, Juliana Kissiedu, Ibrahim A. Hanouneh, Ryan Klatte, 

Charles M. Miller, Nizar N. Zein


841 

Physical function predicts length of stay (LOS) after liver 

transplant (LT): From the Functional Assessment in Liver 

Transplantation (FrAILT) Study 

Jennifer C. Lai, Kenneth E. Covinsky, Iryna Lobach, Sandy Feng; 

UCSF, San Francisco, CA 

Background: Cirrhosis causes sarcopenia and undernutrition 

resulting in progressive decline in physical function(fxn). The 

impact of physical fxn on outcomes after LT is unknown. Methods: 

Adult LT recipients with ≥90 days(d) post-LT follow up 

underwent outpatient pre-LT tests of physical fxn: gait speed+balance+chair 

stands=composite Short Physical Performance 

Battery (SPPB; range 12=high fxn to 0=low fxn). Cut-offs of 

SPPB≤9(low fxn) and lab LT-MELD≥30 were selected for clinical 

relevance. The 1° outcome was LT length of stay (LOS); 

the 2° outcome was home discharge (vs. acute care or skilled 

nursing facility). Adjusted negative binomial regression evaluated 

LOS with physical fxn; Weibull time-to-event analyses 

associated physical fxn with time to discharge home. Results: 

Included were 171 LT recipients: median age 60y. Median 

[interquartile range(IQR)] lab MELD at testing was 16(13-21) 

and time from physical fxn testing to LT 1.9mos (1.0-3.8). Low 

physical fxn (SPPB≤9) was observed in 36%. Median(IQR) lab 

MELD at LT was 22 (17-31); 30% had lab LT-MELD≥30. Median(IQR) 

LOS for SPPB≤9 and >9 were 9(7-20) and 8(6-14) 

[p9 (81vs.89%; p=0.18). In time-to-event analyses adjusted for 

MELD and age, SPPB≤9 was associated with 38% decreased 

hazard of home discharge (95%CI=9-58%; p=0.02). Conclusions: 

Pre-LT physical fxn is a critical determinant of post-LT LOS 

and discharge home. Our data support the implementation 

of objective measures of physical fxn into clinical practice to 

identify those who may benefit from pre- and post-LT physical 

activity interventions. 

842 

Low serum testosterone predicts outcome in men with 

cirrhosis independent of the MELD score 

Marie Sinclair 1,2 , Adam Shannon 2 , Mathis Grossmann 2 , Peter W. 

Angus 1,2 , Rudolf Hoermann 2 , Paul Gow 1,2 ; 1 Gastroenterology, 

Austin Hospital, Heidelberg, VIC, Australia; 2 Medicine, The University 

of Melbourne, Melbourne, VIC, Australia 

Background: Low serum testosterone has been independently 

associated with mortality in a retrospective study of men 

with advanced liver disease. This study aims to prospectively 

explore the relationship between low testosterone and outcome 

in a large cohort of men with cirrhosis Methods: We conducted 

a prospective observational study at a single centre. All men 

with cirrhosis were invited to undergo baseline sex hormone 

profile screening during outpatient consultations. Standard 

parameters including MELD score and Child Pugh Score were 

also documented at baseline. Patients were then followed for 

12 months to assess the composite end-point of mortality or 

liver transplantation. Secondary outcomes include mortality, 

transplantation, hospitalisation and fracture. Results: 203 men 

with cirrhosis were enrolled between April 2013 and March 

2014. The median age was 55 years [50, 64], Child Pugh 

score was 6 [5, 9] MELD score was 10 [8, 15]. The median 

total testosterone level was 17.6nmol/L [8.55, 25.35] (normal 

10-27.6nmol/L) and 28.1% of men had low total testosterone. 

During the 12 month follow-up period 27 patients died and 12 

patients received a liver transplant. Median serum testosterone 

was lower in men who died than those who did not (3.20nmol/L 

[1.05; 12.6] versus 18.1nmol/L [10.6; 26.0], p


safety improvements for these patients. Historically, in-depth 

reviews of medical records were needed to identify AEs, usually 

prompted by a sentinel event. This is very time-consuming, 

requiring a high level of skill and resources. The IHI Global 

Trigger Tool (GTT) helps identify AEs more efficiently, but lacks 

clarity of definitions of complications and guidance for meaningful 

interpretation. We developed a modified GTT to improve 

precision and is simple to use for review of every LD at LDLT 

programs. METHODOLOGY: Medical records were collected 

from 4 Adult to Adult Living Donor Liver Transplantation Cohort 

Study (A2ALL) centers from 2008-2010. Records included all 

documents from admission to discharge and any readmission 

within 30 days after LDLT. A multidisciplinary team (transplant 

surgeon, physician scientist, nurses, and medical student) modified 

the GTT to include transplant specific AEs and created evidence-based 

precise clinical definitions of outcomes, compiled 

into a codebook. Records were analyzed by two reviewers. 

RESULTS: 91 living donor medical records were reviewed. The 

mean age was 39. 57% were male, 77% were Caucasian, 

19% were Hispanic or black, 93% were English speaking. 

722 AEs were identified in 91 records (mean 7.8/patient 

(2-19)). 29% had 5-7 AEs and 14% had 12-14 AEs. The ten 

most frequent AEs were: nausea (68%), constipation (67%), 

hypoxia (64%), anemia (46%), respiratory depression (46%), 

tachypnea (46%), hypotension (42%), atelectasis (41%), pleural 

effusion (41%), fever (37%). With the enhanced GTT, the 

time per for each review was 60 min. Inter-rater reliability was 

> 0.9. DISCUSSION: AEs are far more prevalent than reported 

to date and can highlight areas requiring improvement before 

a sentinel event occurs. The enhanced GTT is a simple and 

efficient tool to identify AE in all living liver donors performed 

a LDLT center and it can be reliably used by anybody (e.g. 

coordinator) after minimal training. 

Disclosures: 

James V. Guarrera - Grant/Research Support: Organ Recovery Systems 

The following authors have nothing to disclose: Blake Platt, Donna Woods, Amna 

Daud, Elizabeth A. Pomfret, Mary Ann Simpson, Robert A. Fisher, Anton I. Skaro, 

Timothy Curtis, Ella F. Reyes, Erin Wymore, Daniela Ladner 

844 

Older Liver Transplant (LT) Candidates with Prolonged 

Wait-Times Have a Low Probability of LT 

Connie W. Wang, Jennifer C. Lai; University of California, San 

Francisco, San Francisco, CA 

Background: The growing wave of older adults with cirrhosis 

has increased demand for LT in ≥65 year(y) olds. While the 

decision to list a patient(pt) for LT may initially be straightforward, 

this decision is particularly dynamic for pts ≥65y who 

are vulnerable to the effects of aging, comorbidities, and even 

decreasing motivation for surgery over time. We aimed to evaluate 

the probability of LT over time among ≥65y pts. Methods: 

All US adult LT candidates listed without MELD exception 

points from 2/02-2/12 were categorized by wait-times


were more likely to die from technical/operative complications 

or multi-organ failure/hemorrhage than “Normal.” Pre-transplant 

functional status is a reliable predictor of postoperative 

mortality for US liver transplant recipients. 

Disclosures: 

The following authors have nothing to disclose: Natasha Dolgin, Paulo N. Martins, 

Adel Bozorgzadeh 

846 

Recanalization of complete anastomotic biliary obstruction 

after liver transplantation by using a through the 

scope magnet: A case series 

Erkan Parlak 1 , Fahrettin Küçükay 2 , Aydin Koksal 1 , Ahmet T. Eminler 

1 , Mustafa I. Uslan 1 ; 1 Gastroenterology, Sakarya University, 

Faculty of Medicine, Sakarya, Turkey; 2 Radiology, Yüksek Ihtisas 

Hospital, Ankara, Turkey 

Aim: It is necessary to traverse a guidewire through the anastomotic 

biliary obstructions after liver transplantation in order to 

perform an endoscopic and/or percutaneous treatment. Otherwise 

the patient either undergoes a surgical revision or lives 

with an external biliary drainage catheter. Herein we evaluated 

the efficacy of a novel ‘’through the scope (TTS)’’ magnet 

in patients with anastomotic biliary obstructions. Methods: 

The magnetic compression anastomotic (MCA) technique: A 

‘’Ni coated cylindirical Neodymium-Iron-Boron rare earth magnet’’with 

a 5 mm length and 2.4 mm diameter was advanced 

over a guidewire to the proximal site of the obstruction via percutaneous 

route and to the distal site of the obstruction through 

a duodenoscope (Figure). Magnets were removed after recanalization 

was achieved. The patients were put on a stent 

exchange programme. Results: One hundred and ten patients 

(93 LRLT, 17 OLT) with biliary anastomotic stricture underwent 

ERCP. TTS MCA was performed in 7 of them (5 LRLT, 2 OLT). 

The procedure was unsuccesfull in 2 LRLT patients with dual 

anastomosis because of long strictures. The procedure was 

successfull in the remaining 5 patients. All of them had a priorly 

placed external biliary drainage catheter. Recanalization was 

achieved after a mean duration of 9 days (range:5-14). No 

magnet related complications were observed. Conclusions: TTS 

MCA is effective and safe in the treatment of complete anastomotic 

biliary obstructions after liver transplantation. Advantages 

and disadvantages with respect to previously defined 

larger magnets should be evaluated in future studies in order to 

determine its indications. 

Disclosures: 

The following authors have nothing to disclose: Erkan Parlak, Fahrettin Küçükay, 

Aydin Koksal, Ahmet T. Eminler, Mustafa I. Uslan 

847 

Patient Safety in Living Donor Liver Transplantation – 

We have a lot to learn 

Daniela Ladner 1 , Elizabeth A. Pomfret 2 , Mary Ann Simpson 2 , 

James V. Guarrera 3 , Robert A. Fisher 4 , Anton I. Skaro 1 , Rebeca 

Khorzad 1 , Ella F. Reyes 1 , Amna Daud 1 , Erin Wymore 1 , Andy 

Hung-Yi Lee 1 , Donna Woods 1 ; 1 CHS, Northwestern University, 

Chicago, IL; 2 Lahey Medical Center, Boston, MA; 3 Columbia Medical 

Center, New York, NY; 4 Virginia Commonwealth University, 

Richmond, VA 

Background: Living donor hepatectomies (LDH) are amongst 

the most scrutinized surgeries by clinicians and patients alike. 

Hence, LDH are probably amongst the most streamlined surgeries 

performed today. To examine the streamlining we focused 

on equipment, an essential element to LDH. Equipment malfunction 

(EM), has been reported in other settings to lead to patient 

injury and in fact, 6% of deaths in hospitalized patients were 

found to be related to equipment errors. Methods: Direct or 

video observation were conducted at four large LDLT centers 

(Feb 2012 - March 2014). Web-based Safety (WBS) Debriefings 

were sent to every clinician (e.g. nursing, physician, technician) 

who participated in an LDH. Results: 110 LDHs were 

debriefed and 22% of all debriefings, reported issues related 

to equipment. Of those 21% related to incorrect, incomplete or 

unavailable instruments, 17% problems related to disposable 

accessories, 7% with surgical lights/monitors and 7% with surgical 

bed (missing/not working). 34 LDH which were directly 

or video observed, demonstrated equipment related issues in 

100% of the cases. Similar issues were observed as reported 

through WBS, but in addition provider’s frustration, lack of


tidiness, and lack of standardization were observed in 9% of 

cases leading to patient injury. Conclusions: EMs happen in 

100% of observed LDHs, one of the most scrutinized surgeries 

performed. While LDH is extremely complex, the flawless 

function of equipment is not. We are far behind other industry 

standards in terms of equipment safety (and other safety), and 

as a medical community have exhibited unparalleled tolerance 

for safety concern even in our most scrutinized and treasured 

LDH. To improve patient safety we need to rethink how we 

organize and execute surgical procedures and reorganize the 

environment to optimally support the execution of the surgical 

procedure. 

Disclosures: 

James V. Guarrera - Grant/Research Support: Organ Recovery Systems 

The following authors have nothing to disclose: Daniela Ladner, Elizabeth A. 

Pomfret, Mary Ann Simpson, Robert A. Fisher, Anton I. Skaro, Rebeca Khorzad, 

Ella F. Reyes, Amna Daud, Erin Wymore, Andy Hung-Yi Lee, Donna Woods 

848 

The effect of recipient and donor diabetes on long-term 

outcomes following liver transplantation, analyzed 

according to underlying disease etiology 

Oliver J. Duncan, Leon A. Adams, Gerry C. MacQuillan, Gary P. 

Jeffrey, George Garas; Hepatology, Sir Charles Gairdner Hospital, 

Perth, WA, Australia 

Background and aims Pre-transplant recipient diabetes and 

donor diabetes have been shown to be independent predictors 

of reduced survival following liver transplantation. The aim 

of this study is to determine the effect of the combination of 

pre-transplant recipient diabetes and donor diabetes on patient 

and graft survival, according to the underlying liver disease. 

Methods Adults (>18 years) who received a liver transplant 

in the United States between 2005 and 2010 were identified 

from the United Network for Organ Sharing registry. Exclusion 

criteria included living related transplant, ABO incompatible, 

multi-organ transplant, donation after cardiac death, 

retransplantation and acute liver failure. We examined the 

impact of donor diabetes and pre-transplant recipient diabetes 

on 30 day, one-year and five-year patient and graft survival 

with subanalysis according to the underlying etiology of liver 

disease. Results Of the 25280 liver transplant recipients identified, 

6053 (23.9%) recipients had a diagnosis of diabetes and 

2849 (11.3%) of the grafts originated from a diabetic donor. 

A total of 705 (2.8%) recipients were diabetic and received 

a liver from a diabetic donor. The commonest indications for 

transplantation included hepatitis C (HCV) (31%), hepatocellular 

carcinoma (HCC) (22.4%) and alcohol (12.4%). The 

median (range) of follow-up was 4.4 (0-9.4) years. The 5-year 

overall patient and graft survival was 72% and 70% respectively. 

Recipient and donor diabetes independently reduced 

5-year survival (69.6% vs. 74% Adjusted Hazard Ratio 1.193, 

95% CI 1.126-1.264 p


Disclosures: 

The following authors have nothing to disclose: Daniela Ladner, Kathryn Skibba, 

Kathryn Jackson, Niloufar Safaeinili, David S. Goldberg, Lisa B. VanWagner, 

Satyender Goel, Abel Kho, Amna Daud, Nazanin Salehitezangi, Anton I. Skaro 

850 

Objective Measurement of Karnofsky Performance Status 

in End Stage Liver Disease Patients 

Lisa Louwers, Emmanouil Palaios, Galal El-Gazzaz, Mario Spaggiari, 

Bijan Eghtesad, Dympna Kelly; Transplantation and Hepatobiliary 

Surgery, Cleveland Clinic, Cleveland, OH 

Purpose In OLT the Karnofsky performance score (KPS) is a 

heavily weighted component in risk stratification in the Scientific 

Registry of Transplant Recipients (SRTR), but there are no 

guidelines as to its use in patients with ESLD. Our goal was to 

develop an objective designation of the KPS in ESLD patients. 

Methods We developed and implemented an algorithm to 

objectively assign KPS to OLT patients, used at the time of OLT 

offer (see figure). We reviewed KPS in our patients before and 

after the tool was implemented. Group 1: 2004 – 2009, 788 

patients, 69% male, mean age 53±13 yrs. Group 2: 2011- 

2014, 468 patients, 68% male, mean age 53±16 yrs. Group 

2 patients had a higher mean chemical MELD score (18±8 vs 

20±11, p=0.001), were more often hospitalized at the time 

of transplant (17.5% vs 23.1%, p=0.024), had a higher incidence 

of COPD and tobacco use (2.6% vs 8.8% and 44.7% 

vs 54.5% respectively, p


of the ≥80y livers (range: 22-65 livers per OPO; “high-yield”). 

Of these high-yield OPOs, 5 of 6 were high-volume OPOs, 

with ≥80y donors accounting for 1.2-2.9% of their total donor 

volume. Discard rates of ≥80y procured livers were similar 

between the 6 high-yield OPOs and all other OPOs that procured 

at least one ≥80y liver (24 vs 26%, p=0.86). Conclusion: 

The majority of oldest old (≥80y) donor livers originated from 

6 high-yield OPOs. These OPOs increased their procured liver 

volume by up to 3% using ≥80y livers, without an increased 

rate of ≥80y liver discard. Developing algorithms to strategically 

evaluate and guide pursuit of the oldest old donors may 

improve efficiency and yield in this rapidly growing population. 

Disclosures: 

Sandy Feng - Consulting: Novartis 

The following authors have nothing to disclose: Suzanne R. Sharpton, Dorry L. 

Segev, Jennifer C. Lai 

Disclosures: 

Jean-Frédéric Blanc - Advisory Committees or Review Panels: Lilly / Imclone, 

Merck Serono, Sanofi; Speaking and Teaching: Bayer, BMS, Roche, Abbvie 

Masatoshi Kudo - Advisory Committees or Review Panels: Bayer HealthCare; 

Grant/Research Support: Bayer HealthCare 




Morris Sherman - Consulting: Gilead, Merck, Bayer, Arqule, Celsion, Boehringer 

Ingelheim, Janssen, Abbvie; Grant/Research Support: WAKO; Speaking and 

Teaching: WAKO 

The following authors have nothing to disclose: Sasan Roayaie, Ghalib Jibara, 

Sarah Berhane, Parissa Tabrizian, Joong-Won Park, Jijin Yang, Lunan Yan, Guohong 

Han, Francesco Izzo, Minshan Chen, Phillip Johnson 

852 

“Oldest Old” (≥80 years) Donors in Liver Transplantation: 

National U.S. Organ Procurement Organization 

(OPO) Practices 

Suzanne R. Sharpton 1 , Sandy Feng 2 , Dorry L. Segev 3 , Jennifer C. 

Lai 1 ; 1 Medicine, University of California, San Francisco, San Francisco, 

CA; 2 Surgery, University of California, San Francisco, San 

Francisco, CA; 3 Surgery, Johns Hopkins, Baltimore, MD 

Background: Reports from non-U.S. single centers have shown 

that increased utilization of the “oldest old” donor livers [≥80 

years (y)] can significantly expand the donor pool. Little is 

known about U.S. OPO practice patterns surrounding the 

oldest old donors. Screening criteria to identify ≥80y donors 

most likely to yield organs are lacking. Methods: We examined 

all U.S. adult deceased donors from 2/2005 to 1/2014 

(n=63,550). OPOs (n=58) were categorized as low-, mid-, 

or high-volume based on tertiles of the total number of donors 

pursued for donation. Logistic regression models investigated 

donor and OPO factors associated with ≥80y liver discard. 

Results: From 403 ≥80y donors, 389 (97%) livers were procured, 

of which 291/389 (75%) were transplanted. Donors 

≥80y vs 2 drinks per day; 6 vs 16%), median AST/ALT (36/23 vs 

45/34), % with liver biopsy (68 vs 40%), and % macrosteatosis 

≥30% (5 vs 15%) [p


854 

Outcomes of Patients Undergoing Liver Transplantation 

using Donors with Confirmed Positive Blood Cultures: A 

review of the UNOS dataset 

Moises Huaman 2 , Valery Vilchez 1 , Xiaonan Mei 1 , Malay B. 

Shah 1 , Michael F. Daily 1 , Roberto Gedaly 1 ; 1 Surgery, Univ of Kentucky, 

Lexington, KY; 2 Division of Infectious Diseases, Department 

of Medicin, University of Kentucky, Lexington, KY 

Background: The potential effect of blood culture positive donor 

(BCPD) on liver transplant outcomes has not been well established. 

We aimed to evaluate the impact of BCPD in patient 

and graft survival after liver transplantation. Methods: We 

retrieved data from the United Network for Organ Sharing 

(UNOS) registry on all adults who underwent primary, single 

organ deceased-donor liver transplantation in the US between 

2008 and 2013. Patients were classified in two cohorts: the 

BCPD cohort and the non-BCPD cohort. Graft and patient survival 

at 30 days and 1 year were compared between cohorts. 

Multivariable analysis using Cox Proportional Hazard models 

for graft survival was performed. Results: A total of 28,961 

patients were included in the analysis. There were 2,316 

(8.0%) recipients of BCPD. BCPD were more likely to received 

organs from female diabetic donors with higher body mass 

index compared to non-BCPD. Although patient survival was 

similar at 7 days, 1-, 6- and 12 months we found a significant 

difference in graft survival at this time lines (p=0.009). On multivariable 

analysis, BCPD was independently associated with 

decreased graft survival (adjusted OR; 1.10, 95% CI 1.005 

– 1.203; P


scores 6-19 or 30-40 did not vary with BMI (47% and 77% 

post-LT mortality reduction, respectively, regardless of BMI). 

Conclusion: Low BMI is associated with decreased LT survival 

benefit in patients with moderate MELD scores. If validated, 

these results support the consideration of BMI during LT listing. 

significantly increased as LT survival benefit decreased. ROC 

analysis showed excellent accuracy of LT futility prediction in 

both cohorts. Conclusion: We describe a scoring system that 

predicts LT futility based on LT survival benefit decrement. This 

has the potential to improve organ allocation. 

Association of BMI with LT survival benefit by MELD scores 

Disclosures: 

The following authors have nothing to disclose: Mohannad Dugum, Nizar N. 

Zein, Alex Dugum, Rocio Lopez, Bijan Eghtesad, Ibrahim A. Hanouneh 

856 

Predicting Futility of Liver Transplantation (LT): A Scoring 

System Based on LT Survival Benefit 

Mohannad Dugum 1,2 , Nizar N. Zein 3 , Alex Dugum 4 , Rocio 

Lopez 5 , Bijan Eghtesad 6 , Ibrahim A. Hanouneh 3 ; 1 Internal Medicine, 

Cleveland Clinic, Cleveland, OH; 2 Gastroenterology, 

Hepatology and Nutrition, University of Pittsburgh, Pittsburgh, PA; 

3 Gastroenterology and Hepatology, Cleveland Clinic, Cleveland, 

OH; 4 University of California, San Diego, San Diego, CA; 5 Quantitative 

Health Sciences, Cleveland Clinic, Cleveland, OH; 6 Transplantation 

Center, Cleveland Clinic, Cleveland, OH 

Background and aim: Assessment of LT futility is essential to 

improve organ allocation. We aimed to devise a scoring system 

to predict LT futility based on factors affecting LT survival benefit. 

Methods: We defined LT survival benefit as the difference 

between life expectancy if transplanted and life expectancy if 

the patient remains on the waiting list (WL). LT was considered 

futile if the LT survival benefit was negative. Adult patients listed 

for LT in the US 1987-2012 were identified from the UNOS 

database. After excluding malignant diagnoses and status 1, 

patients were randomly divided to 2 cohorts: 2/3 training 

(for model building) and 1/3 validation. Pre-LT survival was 

modeled using competing risks analysis and post-LT survival 

was assessed using Cox regression. Using these models, life 

expectancy on WL and post-LT was estimated for each patient 

by calculating the area under survival curve up to 5 years using 

the trapezoidal rule. Using the training cohort, multivariable 

logistic regression analysis was performed to build a model 

for prediction of LT futility. Multiple factors were considered 

for inclusion in the model and backward elimination was used 

to remove factors with p>0.05. The model was then applied 

in the validation cohort, and ROC analysis was performed to 

assess the accuracy of LT futility prediction. Results: 33,376 

patients were included in the training cohort. Average age 

at listing was 53±10 years and 44% underwent LT. Median 

LT survival benefit was 6.5 months [0.2, 19.1]. Older age 

at listing, higher BMI, lower serum sodium, and higher MELD 

predicted survival. Similar results were observed in the LT population. 

The proposed score consists of age, BMI, serum sodium 

and MELD. When applied in the validation cohort, the score 

Disclosures: 

The following authors have nothing to disclose: Mohannad Dugum, Nizar N. 

Zein, Alex Dugum, Rocio Lopez, Bijan Eghtesad, Ibrahim A. Hanouneh 

857 

Remnant Liver Ischemia Is Associated With Early Recurrence 

and Poorer Survival after Liver Resection 

Jai Young Cho, Ho Seong Han, YoungRok Choi, Yoo-Seok Yoon, 

Jae Yool Jang, Hanlim Choi, Jae Seong Jang, Seong Uk Kwon; 

Seoul National University Bundang Hospital, Seongnam, Korea 


Background: Non-anatomical hepatectomy or compromised 

blood supply to the remnant liver may result in remnant liver 

ischemia (RLI) of various extension and severity. There are a 

few reports showing association between ischemia-reperfusion 

injury and tumor recurrence in animal liver transplantation 

model. However, there is no report to evaluate the impact of 

RLI after hepatectomy for hepatocellular carcinoma (HCC) on 

patient survival. Method: Remnant liver ischemia was graded 

on postoperative CT scan in 328 patients who underwent 

hepatectomy for HCC between January 2004 and December 

2013. We defined RLI as reduced or absent contrast enhancement 

during the venous phase and classified to minimal (none 

or marginal) or significant (partial, segmental, and necrosis). 

Results: We observed radiologic signs of significant RLI in 98 

patients (29.9%): 63 partial, 16 segmental, and 19 necrosis, 

and these patients showed more complications (P < 0.001) 

and longer hospital stays (P = 0.002). Preoperative history of 

transarterial embolization (P = 0.040), use of Pringle maneuver 

(P = 0.028), and longer operation time (P < 0.001) were independent 

risk factors for developing RLI. Patients with significant 

RLI showed higher rates of early recurrence within 6 or 12 

months after hepatectomy compared those without (P < 0.001). 

Moreover, RLI was independent risk factors for both overall 

patient (P < 0.001; RR = 6.984; 95% CI, 4.268-11.426) and 

disease-free survival (P < 0.001; RR = 5.153; 95% CI, 3.615- 

7.345). Conclusion: Partial hepatectomy without RLI and medical 

treatment for RLI are highly recommended in patients with 

HCC. 

Disclosures: 

The following authors have nothing to disclose: Jai Young Cho, Ho Seong Han, 

YoungRok Choi, Yoo-Seok Yoon, Jae Yool Jang, Hanlim Choi, Jae Seong Jang, 

Seong Uk Kwon


858 

Novel Sonar Guided Technique For Management Of 

Incarcerated Umbilical Hernia In Cirrhotic Patients 

Awaiting Liver Transplantation 

Abd Elrazek A. Hussein 1 , Shymaa E. Bilasy 2 , Hamdy M. Moustafa 1 , 

Mohammed Nafady 3 ; 1 Hepatology and Liver Transplantation, Al 

Azhar School of Medicine, Asuit, Egypt; 2 Biochemistry, Faculty of 

Pharmacy, Suez Canal University, Ismalia, Egypt; 3 Radiology, Al 

Azhar Faculty of Medicine, Asuit, Egypt 

Background and Aim: Umbilical hernias pose a management 

dilemma in patients with cirrhosis, especially those awaiting 

liver transplantation. Complications of umbilical hernias in cirrhotic 

patients with ascites include leakage, ulceration, rupture 

and incarceration thus leading to a higher mortality rate after 

surgical repair. Hernias are usually repaired at the time of 

transplantation. However, as the waiting time prior to liver 

transplantation is prolonged, the likelihood of an incarcerated 

hernia developing is increased. Here, we investigated the feasibility 

of using sonar guided repair for incarcerated hernia. 

Patients and Methods: This study included 23 cirrhotic patients 

examined at the centers of GIT, Hepatology, Radiology, 

Faculty of Medicine- Al Azhar University- Egypt. All patients 

showed the clinical manifestations of incarceration particularly, 

tense feeling of the hernia and manifestation of acute intestinal 

obstruction. Both conventional and color doppler abdominal 

sonography revealed the extent of obstruction and the vasculature. 

Three essential steps were followed to repair the hernia 

under ultrasonography guidance. First, we preformed complete 

aspiration of the ascetic fluid and the fluid inside the hernia 

sac. Then, the distended intestine inside the sac was deflated 

to some extent by syringe needle of 16-18 gauge which was 

introduced obliquely through the intestinal wall under sonography 

guidance. Finally, the hernia was completely reduced 

by gentle compression upon the hernia using sterilized jell as 

a lubricant and cold fomentation to decrease the size of the 

herniated sac. After reduction of the hernia contents, a pressure 

pad of dressing was applied at the site of the hernia and 

fixed with adhesive tapes to prevent immediate recurrence. 

The patient was informed to leave the pressure dressing in situ 

and avoid any straining. Results: This study included 20 males 

and 3 females between the age of 40-70 yr with incarcerated 

hernia between 5 – 9 cm and incarceration duration between 

6 – 30 hours. Color doppler sonography revealed inappropriate 

intestinal vasculature in three cases and those were referred 

back to their surgeons for immediate operation. The remaining 

20 patients, showed appropriate intestinal vasculature. Using 

the above mentioned sonar guided technique, we succeeded 

in repairing hernia in 17 patients (7male and 10 female) with 

overall success rate of 73.9%. Conclusion: To our knowledge, 

this is the first report to describe sonar guided manipulation 

for fixing incarcerated umbilical hernia. Thus, this described 

procedure can be either a safe alternative to surgery or at least 

as a bridge for later safe elective surgery. 

Disclosures: 

The following authors have nothing to disclose: Abd Elrazek A. Hussein, Shymaa 

E. Bilasy, Hamdy M. Moustafa, Mohammed Nafady 

859 

Safety of live liver donation by individuals with G6PD 

deficiency: Initial results and comparative study 

Mettu S. Reddy, Manoj V. Shrivastav, Ilankumaran Kaliamoorthy, 

Mohamed Rela; Institute of liver disease and transplantation, 

Global health city, Chennai, India 

Background G6PD deficiency is the most common genetic 

human enzyme defect in the world. Baring a single case report, 

there is no published literature regarding the safety of donor 

hepatectomy in G6PD deficient individuals. We established a 

protocol of selective utilization of G6PD deficient liver donors in 

our LDLT program in September 2013. This manuscript presents 

the initial results ofour donors with G6PD deficiency (G6PDd). 

Methods All potential donors underwent qualitative and quantitative 

assessment of G6PDd. Donors with WHO Class III or 

Class IV G6PDdwere evaluated for donation if there was no 

other suitable donor in the recipient’s family and pre-operative 

screening revealed no evidence of previous or ongoing haemolysis. 

Care was taken to avoid medications which induce 

haemolysis in the peri-operative period. Donors were monitored 

for haemolysis. Post-operative course of these 14 patients was compared 

with a cohort of 30 non-G6PDd donors case-matched 

for age, BMI, type of graft and residual liver volume.> 

Results Between September 2013 and May 2015, 14 donors 

with G6PDd underwent donor hepatectomy. There were 3 right 

lobe, 2 left lobe and 9 left lateral segment donors. No donor 

received intra-operative transfusion. Two donors with G6PDd 

had biochemical evidence of post-operative haemolysis which 

was self-limiting. No donor either in the G6PDd or non-G6PDd 

group had greater than Clavien 3A complication. Post-op liver 

function tests, ICU stay (4 vs 3.5 days, p=0.117), hospital 

stay (9 vs 9 days, p=0.517) and % with greater than Clavien 

II grading was similar in the two groups (7.1% vs 6.6%, 

p=0.187). Donors in the G6PDd group had similar pre-op haemoglobin 

(12.4g% vs 12.9g%, p=0.404) but had lower trough 

haemoglobin in post-op period (8.1g% vs 92g%, p=0.006), 

greater drop in post-op haemoglobin (33.6% vs 25.7%, p= 

0.007) and a higher need for post-op blood transfusion (4/14 

vs 2/30, p=0.071). Conclusions We present the first reported 

series of 14 G6PD deficient individuals who underwent donor 

hepatectomy. G6PDd donors can have self-limiting haemolysis 

post-operatively and may have a slightly greater need for 

post-operative transfusion. Donor hepatectomy can be carried 

out safely in carefully selected donors with G6PD deficiency as 

long as they are managed in a vigilant clinical setting. 

Disclosures: 

The following authors have nothing to disclose: Mettu S. Reddy, Manoj V. Shrivastav, 

Ilankumaran Kaliamoorthy, Mohamed Rela


860 

Living donor liver transplantation for patients with HCC 

exceeding Milan criteria: results of pilot study. 

Josep M. Llovet 1,2 , Mihai-Calin Pavel 3 , Jordi Rimola 1 , Maria 

Alba Diaz 4 , Jordi Colmenero 3,5 , Constantino Fondevila 3 , Josep 

Fuster 1,3 , Pere Gines 3,5 , Jordi Bruix 1,5 , Juan Carlos García-Valdecasas 

3 ; 1 Liver Unit, BCLC, IDIBAPS, CIBEREHD, Universitat de Barcelona, 

Barcelona, Spain; 2 Institució Catalana de Recerca i Estudis 

Avançats, Barcelona, Spain; 3 Liver Transplant Unit, Digestive and 

Metabolic Diseases Institute, Hospital Clinic, University of Barcelona, 

Barcelona, Spain; 4 Pathology Department, Hospital Clinic, 

University of Barcelona, Barcelona, Spain; 5 Liver Unit, Digestive 

and Metabolic Diseases Institute, Hospital Clinic, University of Barcelona, 


Introduction: Liver transplantation is the standard of care for 

patients with hepatocellular carcinoma (HCC) within Milan criteria 

not suitable for resection. Nonetheless, during the last 

20 years it became apparent that a subset of patients beyond 

Milan criteria might obtain acceptable survival outcomes. In 

parallel, living donor liver transplantation has emerged as a 

feasible alternative to overcome the paucity of donors. Methods: 

In 2002, we proposed for LDLT in Child A-B patients with 

HCC a set of criteria that substantially expanded the conventional 

indications of transplantation (1 tumor up to 7cm, 5 

tumors of < 3cm, 3 tumors < 5cm, or downstaging to Milan 

lasting 6 mo after loco-regional therapies) (Bruix& Llovet, Hepatology 

2002). Results: We herein present a prospective cohort 

of 22 patients with HCC fulfilling these criteria that were treated 

with LDLT between 2002 and 2014. Median age was 57 yrs, 

20 men, HCV related 14, Child-Pugh A:16, B:6, AFP


862 

Clinical impact of 18 F-FDG-PET/CT in living donor liver 

transplantation for advanced hepatocellular carcinoma 

Seung Duk Lee, Eung Chang Lee, Seong Hoon Kim; Liver Cancer 

Center, National Cancer Center, Republic of Korea, Goyang-si, 


Background: The relevant number of patients with hepatocellular 

carcinoma (HCC) beyond the Milan criteria has undergone 

living donor liver transplantation (LDLT). However, the prognostic 

factors for these patients with advanced HCC are not 

well established. Methods: From March 2005 to May 2013, 

280 patients with HCC underwent LDLT in the National Cancer 

Center. Among these, patients beyond the Milan criteria 

were retrospectively enrolled. We analyzed the prognostic significance 

of 18 F-fluorodeoxyglucose positron emission tomography/computed 

tomography ( 18 F-FDG-PET/CT) for selecting 

appropriate candidates. Results: Among total 280 patients, 

147 patients (52.5%) were confirmed to have HCC beyond 

the Milan criteria using pathological reports. The patients who 

met and exceeded the Milan criteria had 5-year overall survival 

(OS) rates of 87.2% and 64.6%, respectively, (p < 0.001). In 

multivariable analysis for OS and DFS in patients beyond the 

Milan criteria, PET/CT positivity [hazard ratio (HR) 2.714, p 

= 0.013 for OS; HR 3.803, p < 0.001 for DFS], total tumor 

size over 10 cm (HR 2.333, p = 0.035 for OS; HR 3.334, p 

= 0.001 for DFS), and microvascular invasion (HR 2.917, p = 

0.025 for DFS) were significant prognostic factors. In particular, 

patients beyond the Milan criteria with a PET/CT-negative 

status and total tumor size < 10 cm showed similar OS and 

DFS in comparison with those within the Milan criteria. Conclusions: 

A PET/CT status in LDLT is a useful marker to predict 

survival of patients with advanced HCC. 

Disclosures: 

The following authors have nothing to disclose: Seung Duk Lee, Eung Chang Lee, 

Seong Hoon Kim 

863 

Living or Deceased Donor Liver Transplantation for 

Hepatocellular Carcinoma: A Western multicenter, intention 

to treat, cohort study 

Daniel Azoulay; HPB Surgery and liver transplant, Hopital Mondor 

APHP, Creteil, France 

Purpose. An intention-to-treat analysis of overall survival (ITT-OS) 

in cirrhotic patients with hepatocellular carcinoma (HCC) listed 

for either living donor (LDLT) [Group LiLDLT] or deceased donor 

liver transplantation (DDLT), [Group LiDDLT] across 5 French 

liver transplantation (LT) centers. Methods. Records of 861 cirrhotic 

patients with HCC who were consecutively listed for 

either LDLT (n=79) or DDLT (n=782) from 2000 to 2009 at 5 

French centers were analyzed for ITT-OS using Cox model, and 

for tumor recurrence using a competitive risk model. Results. 

Tumour staging in both groups was similar. Among the listed 

patients, 162 dropped-out (20.7%), all from Group LiDDLT (p < 

0.0001). At 5-years, ITT-OS was significantly better for Group 

LiLDLT vs Group LiDDLT (73.2% vs 66.7%;p = 0.062). Being 

listed for LDLT (HR: 0.62 (0.40-0.98); p = 0.039), and MELD 

score ≥ 25 at listing (HR: 1.79 (1.11-2.88); p = 0.017) were 

independent predictors of ITT-OS. For the 699 transplanted 

patients, 5-year OS post-LT (73.2% and 73.0%, p = 0.407) 

and HCC recurrence (10.9 % and 11.2 %, p = 0.753) were 

similar between those who had a living donor LT [Group LDLT] 

and deceased donor LT [Group DDLT], respectively. Tumor 

beyond Milan criteria (HR = 2.67 (1.5-4.7); p < 0.001), and 

vascular invasion (HR = 2.52 (1.38;4.59); p = 0.003) were 

independent predictors of recurrence whereas the type of LT 

was not. Conclusion. LDLT improves ITT-OS by obviating dropout, 

is not a risk factor of tumor recurrence, and hence should 

be equally encouraged in countries where both, LDLT and DDLT 

are available. 

Disclosures: 

The following authors have nothing to disclose: Daniel Azoulay 

864 

Third Liver Transplantation - Utility and Principles to 

Guide Future Transplants; an Analysis of UNOS data 

Vandana Khungar 1 , David S. Goldberg 1 , Peter L. Abt 2 ; 1 Medicine, 

Division of Gastroenterology, University of Pennsylvania, Philadelphia, 

PA; 2 Surgery, University of Pennsylvania, Philadelphia, PA 

Background: Repeat liver transplantation (reLT), whether for 

recurrent disease, vascular complications, or chronic rejection, 

can be a lifesaving therapy. However, donor livers are 

a scarce resource, with up to 20% of patients dying on the 

waitlist before receiving their first transplant. As more patients 

get further out from their 1 st (or 2 nd ) transplant, the transplant 

community will need to consider the utility of 3 rd liver transplants. 

Yet no data specifically addressing this question exist. 

Methods: We performed a retrospective cohort study of all 

adult patients listed for their third transplant, who were on 

the waiting list between 2/28/02-12/31/2014, using UNOS 

data. Kaplan-Meier analyses and log-rank tests were used to 

evaluate post-transplant outcomes among the subset of patients 

who received their third transplant. Results: 696 adult patients 

were on the waitlist for a 3 rd transplant in this period. 362 

(52.0%) received a 3 rd transplant, while 218 (31.3%) were 

removed from the list for death or clinical deterioration. The 

median recipient age was 47 (IQR: 34-45), median donor 

risk index (DRI) was 1.33 (IQR: 1.15-1.58), with 211 (58.3%) 

donors being under the age of 40. Median laboratory MELD at 

transplant was 29 (21-36). The 1,3, and 5 year post-transplant 

patient survival rates were 74.5%, 64.4%, and 57.2% respectively. 

Graft and patient survival were not statistically different 

between centers who had performed 1-9 3 rd transplants vs 

≥10 3 rd transplants (p>0.05). Post-transplant patient survival 

differed significantly (p=0.001) based on the transplant recipient’s 

location at the time of 3 rd transplant. Conclusions: Patients 

undergoing reLT were given optimal organs from young donors 

with low DRI. Despite this, their post-transplant survival is lower 

when compared to patients undergoing 1 st transplantation. 

The survival rates among the subset transplanted from the ICU 

are dramatically lower, and raise questions about the utility of 

transplantation in these high-risk patients outside of the setting 

of primary non function (PNF) or hepatic artery thrombosis 

(HAT).


Patient survival by location at time of 3rd transplant 

Disclosures: 

The following authors have nothing to disclose: Vandana Khungar, David S. 

Goldberg, Peter L. Abt 

865 

Mechanisms of Gender Disparities in Liver Transplantation: 

An Examination of Organ Offers 

Lauren D. Nephew, James D. Lewis, David S. Goldberg, Kimberly 

A. Forde; Gastroenterology, Hospital of the University of Pennsylvania, 


Background: In the MELD era, women are more likely than men 

to become too sick or die on the liver transplant list and less 

likely than men to receive liver transplantation.Gender differences 

in creatinine, a component of the MELD score, is a potential 

contributing factor. However corrected MELD scores that 

account for these differences have not been shown to improve 

estimates of 3-month mortality or rates of transplantation in 

women. However, receiving a liver transplant is a complex 

process that involves first receiving an organ offer. The role of 

organ offers in gender disparities in liver transplant have yet to 

be explored. We hypothesize that female waitlist candidates 

will have lower MELD scores compared to male waitlist candidates 

and as a result will receive fewer organ offers. Methods: 

A retrospective cohort study of patients listed for liver transplantation 

in the United States was conducted. We included adult 

match runs from May 10, 2007 through June 17, 2013. Given 

different criteria for wait listing and organ allocation for multiorgan 

transplantation, retransplantation, bypasses, and status 

1, these patients were excluded from the analysis. The primary 

outcome was receipt of an organ offer defined as appearing 

in the first position on any match run. Multilevel mixed effects 

logistic regression modeling was used to account for clustering 

of the data by donor service area. Univariable analysis was 

performed to evaluate unadjusted gender differences in the 

odds of receiving an organ offer. Multivariable analysis then 

evaluated region, blood type, race, exception point status, 

etiology of liver disease, and listing MELD as potential confounding 

variables. Results: There were 64,995 candidates 

who appeared on a match run during our study period. Organ 

offers were made to 10,714 candidates (65.1% male, 35% 

female). On univariable analysis, there was no association 

between gender and organ offer (OR=0.98, CI 0.94-1.02). 

When blood type, ethnicity/race, exception point status, etiology 

of liver disease, and region were added to the model, 

their remained no association between gender and organ offer 

(OR=.98, CI 0.94-1.03). Conclusions: We hypothesized that 

lower creatinine values could result in lower MELD scores and 

hence fewer organ offers for female transplant candidates. 

However, female waitlist candidates are just as likely as men 

to receive an organ offer. These analyses suggest that gender 

disparities in liver transplantation do not occur at the level of 

the organ offer. Future work will explore gender disparities in 

organ refusals. 

Disclosures: 

James D. Lewis - Grant/Research Support: Bayer 

The following authors have nothing to disclose: Lauren D. Nephew, David S. 

Goldberg, Kimberly A. Forde 

866 

Significance of functional hepatic resection rate using 

3D fusion image of CT and 99m Tc-galactosyl human 

serum albumin scintigraphy 

Yosuke Tsuruga, Toshiya Kamiyama, Hirofumi Kamachi, Shingo 

Shimada, Kenji Wakayama, Tatsuya Orimo, Hideki Yokoo, Akinobu 

Taketomi; Gastroenterological Surgery I, Hokkaido University 

Graduate School of Medicine, Sapporo, Japan 

Background: Preoperative estimation of hepatic functional 

reserve is important for major hepatectomy. Though future 

remnant hepatic “volume” is calculated by CT images as volumetry, 

it was difficult to estimate the “function” of future liver 

remnant. Hepatic receptor imaging with 99m Tc-galactosyl-human 

serum albumin ( 99m Tc-GSA) scintigraphy is frequently 

used for evaluating hepatic functional reserve. In this study, 

we evaluated the usefulness of calculation of functional hepatic 

resection rate (FHRR) using 3D fusion image of CT and 99m Tc- 

GSA single-photon emission computed tomography (SPECT) 

scintigraphy. Methods: 57 patients who underwent more than 

one sectionectomy at our institution between October 2013 

and March 2015 were enrolled in this study. Of these, 26 

patients presented with hepatocellular carcinoma, 12 with hilar 

cholangiocarcinoma, 6 with intrahepatic cholangiocarcinoma, 

4 with liver metastasis, and 9 with other diseases. 50 patients 

underwent bisectionectomy, and 7 trisectionectomy. We compared 

the volume hepatic resection rate (VHRR) with FHRR. 

FHRR was defined as the resection volume counts per total liver 

volume counts of 99m Tc-GSA SPECT from 3D fusion image with 

CT. Results: FHRR and VHRR were 38.6 ± 19.9 and 44.5 ± 

16.0 (mean ± standard deviation) respectively. The regression 

coefficient of FHRR on VHRR was 1.16 and the coefficient of 

determination was 0.87 (p 5.0mg/dl) was 

observed in 6 of 57 patients. There was no operation-related 

death. Conclusion: Calculation of FHRR was important for 

major hepatectomy because there was a discrepancy between 

FHRR and VHRR in some cases due to hepatic insufficient inflow 

and congestion. 

Disclosures: 

The following authors have nothing to disclose: Yosuke Tsuruga, Toshiya Kamiyama, 

Hirofumi Kamachi, Shingo Shimada, Kenji Wakayama, Tatsuya Orimo, 

Hideki Yokoo, Akinobu Taketomi 

867 

Auxiliary partial orthotopic liver transplantation (APOLT) 

for metabolic liver disease- is it still relevant? 

Mohamed Rela 2 , Priya Ramachandran 1 , Mohamed Safwan 2 , 

Naresh P. Shanmugam 2 , Sanjay Govil 2 , Mettu S. Reddy 2 ; 1 Pediatric 

Surgery, CHILDS Trust Hospital, Chennai, India; 2 Institute of 

Liver Disease and Transplantation, Global Hospital & Health City, 

Chennai, India 

Purpose: The role of auxillary partial orthotopic liver transplantation 

(APOLT) for non-cirrhotic metabolic liver disease 

(MLD) is controversial because of greater technical challenges 

and higher rejection rates when compared to orthotopic liver 

transplantation (OLT). The aim of our study was to examine 

the outcome of APOLT in our series of patients with selected 

non-cirrhotic MLD. Methods: We retrospectively reviewed the 

case notes of all patients who underwent APOLT for MLD in our


centre. Operative details, post-operative course and current 

status of these patients were reviewed. Results: Over a 5 year 

period (2010-2015), 13 patients underwent APOLT in our centre. 

Of these, 8 patients (5 males) underwent APOLT for MLD. 

The indications were Propionic acidemia (PA) (n=4), Citrullinemia 

(n=1) and Crigler-Najjar syndrome (CNS) (n=3). The 

median age at transplantation for PA and citrullinemia was 38 

months (9-52 months) while the median age for patients with 

CNS was 14 years (6-23 years). Five patients received living 

related donor grafts, two (1 Citrullinemia and 1 CNS) received 

deceased donor grafts, and one child with CNS received a 

partial left lobe graft recovered from the child with PA who 

underwent a left lobe APOLT (Domino graft). Graded portal 

vein banding (to prevent portal steal to the graft) was done 

in all cases. One child had hepatic artery thrombosis on day 

4 which was successfully managed by thrombectomy. One 

child developed two episodes of acute rejection diagnosed by 

biopsy and managed with steroid pulse therapy. All patients 

are well (median follow up 9 months) (range 14 days-55 

months) with normal liver function tests and unrestricted protein 

intake and activity. Conclusion: The outcome of APOLT in our 

series is comparable to the reported outcomes for OLT in MLD. 

APOLT is an acceptable treatment for non-cirrhotic MLD. The 

major advantages are the safety of the remnant native liver in 

case of technical complications which can translate into better 

patient survival and the lack of an anhepatic phase, which 

ensures a smooth intra-operative period. In addition the patient 

will have the option of future corrective therapies in the native 

liver. 

Disclosures: 

The following authors have nothing to disclose: Mohamed Rela, Priya Ramachandran, 

Mohamed Safwan, Naresh P. Shanmugam, Sanjay Govil, Mettu S. Reddy 

868 

Graft Survival following Domino Liver Transplantation: 

The U.S. Experience 

Ian W. Folkert 1 , David S. Goldberg 2 , Matthew H. Levine 1 , Kim M. 

Olthoff 1 , Abraham Shaked 1 , Peter L. Abt 1 ; 1 Surgery, Hospital of 

the University of Pennsylvania, Philadelphia, PA; 2 Gastroenterology, 

Hospital of the University of Pennsylvania, Philadelphia, PA 

Introduction: Domino liver transplantation, in which the 

explanted liver from a patient with a hepatic metabolic disorder 

(domino donor - DD) is subsequently transplanted into 

a second patient (domino recipient - DR), has been utilized 

as a method to increase the donor organ supply. Outcomes 

of domino transplantation in the U.S. have not yet been fully 

evaluated, nor compared to recipients of deceased donor and 

living donor grafts. Methods: We performed a retrospective 

review of all domino liver transplants performed in the United 

States utilizing the UNOS liver transplant database. Graft survival 

of DD and DR were compared to recipients of deceased 

donor and living donor liver grafts respectively. Results: From 

2000-2014, there were 144 domino transplants performed at 

38 different centers. Mean DD age was 47; 66% were male. 

Mean DD waitlist time was 276 days. The most common diagnosis 

leading to transplant in the domino donor was familial 

amyloidosis (80%). 26 (18%) DD received multiple organs at 

the time of transplant, with 22 receiving a simultaneous heart 

transplant. Mean age of the DR was 57 years; 67% were male. 

Mean waitlist time for the DR was 543 days. The most common 

diagnosis leading to transplant in the domino recipient was 

hepatitis C (26%). Mean MELD score at time of transplant for 

domino donors vs. deceased donor controls was 8 vs. 21. 

Mean MELD score at time of transplant for domino recipients 

vs. living donor controls was 15 vs. 15. Kaplan-Meier survival 

analysis demonstrated no statistical difference in graft 

survival between domino donors and a control group receiving 

deceased donor grafts at 1 (88% vs. 85%, p=.34) and 5 years 

(71% vs. 69%, p=.56). There was also no difference in graft 

survival between domino recipients and living donor controls at 

1 (85% vs. 83%, p=.57) and 5 years (68% vs. 71%, p=.66). 

Conclusions: Domino transplantation is an effective strategy to 

increase liver transplantation, with post-transplant graft survival 

that is not different when compared to transplantation using 

deceased donor or living donor grafts. 

Disclosures: 

The following authors have nothing to disclose: Ian W. Folkert, David S. Goldberg, 

Matthew H. Levine, Kim M. Olthoff, Abraham Shaked, Peter L. Abt 

869 

Application of Gadoxetate Disodium-Enhanced MR 

Imaging in a Rat Model of Acute Partial Liver Congestion 

Akira Shimizu, Akira Kobayashi, Takahide Yokoyama, Hiroaki 

Motoyama, Hiroshi Sakai, Noriyuki Kitagawa, Tsuyoshi Notake, 

Tomoki Shirota, Kentaro Fukushima, Shinichi Miyagawa; Surgery, 

Shinshu University, School of medicine, Matsumoto, Japan 

Purpose: To evaluate the features of hepatic congestion on 

gadoxetate disodium (Gd-EOB-DTPA)-enhanced magnetic resonance 

imaging (MRI) and the mechanisms responsible for 

the radiological findings in a rat model of acute partial liver 

congestion. Materials and Methods: We used male Wistar rats 

weighing 250–320 g. A conventional T1-weighted spin-echo 

sequence of the liver was performed using a 1.5T magnetic 

resonance imager with an 80-mm magnetic aperture for animal 

studies. We induced regional liver congestion using partial left 

lateral hepatic vein ligation (n = 5) and evaluated the following 

in both congestive liver (CL) and non-congestive liver (non-CL): 

1) chronological changes in the relative enhancement (RE) up 

to 60 minutes after Gd-EOB-DTPA administration and other 

parameters including the maximum RE value (C max 

), time to 

maximum RE (T max 

), elimination half-life of RE (T 1/2 

), and the 

hepatocellular uptake index per unit volume (HUI/V), 2) the 

amount of bile secretion and ATP concentration, and 3) mRNA 

and protein expression of rat organic anion transporting protein 

1a1 (Oatp1a1) using real-time PCR and immunohistological 

examination. Results: 1) The RE in the CL reached a small 

peak (18%) at 5 minutes, corresponding to approximately half 

of the value observed in the non-CL, then slowly decreased in 

a linear manner thereafter. The degree of RE in the CL was 

significantly lower than that in the non-CL for up to 30 minutes 

(P < 0.05). 2) The amount of bile secretion was significantly 

decreased in the CL (3.6 ± 4.1 μL/h/g of liver) compared with 

those in the non-CL (90.2 ± 32.2 μL/h/g of liver, P < 0.001). 

Similarly, the ATP concentration was significantly reduced in 

the CL (0.5 ± 0.2 μmol/h/g wet liver) compared with those in 

the non-CL (3.3 ± 1.3μmol /g wet liver, P < 0.001). There were 

significant correlations between the ATP concentration and the 

HUI/V (r 2 = 0.593, 95% CI 0.587-0.599, P = 0.004), C max 

(0.546, 0.538-0.555, P = 0.008) and T max 

(0.458, 0.487- 

0.500, P = 0.012); the closest correlation was observed with 

the HUI/V. 3) An immunohistological examination showed that 

Oatp1a1 protein expression was downregulated in the CL. 

The mRNA level of Oatp1a1 in the CL was significantly upregulated, 

compared with that in control rat liver (P = 0.046), 

whereas no significant difference was observed between the 

CL and the non-CL (P = 0.698). Conclusion: Gd-EOB-DTPA-enhanced 

MRI is useful for detection of the congestive liver area 

clearly depicted as hypointense. Reduced signal intensity in 

the congestive area on Gd-EOB-DTPA-enhanced MRI could


be explained by the decreased uptake of contrast agent via 

Oatp1a1 protein. 

Disclosures: 

The following authors have nothing to disclose: Akira Shimizu, Akira Kobayashi, 

Takahide Yokoyama, Hiroaki Motoyama, Hiroshi Sakai, Noriyuki Kitagawa, 

Tsuyoshi Notake, Tomoki Shirota, Kentaro Fukushima, Shinichi Miyagawa 

Disclosures: 

Thomas D. Schiano - Advisory Committees or Review Panels: salix, merck, gilead, 

pfizer; Grant/Research Support: galectin, massbiologics, biotest 

The following authors have nothing to disclose: Amy Tan, M. Isabel Fiel, Stephen 

C. Ward, Marcelo Facciuto, Swan N. Thung, Myron E. Schwartz, Sander S. 

Florman 

870 

Lack of Radiologic-Pathologic Correlation and Associated 

Clinical Features of Patients with Hepatocellular 

Carcinoma Undergoing Liver Transplantation 

Amy Tan 1 , M. Isabel Fiel 3 , Stephen C. Ward 3 , Marcelo Facciuto 4 , 

Swan N. Thung 3 , Myron E. Schwartz 4 , Sander S. Florman 4 , 

Thomas D. Schiano 2 ; 1 Internal Medicine, Icahn School of Medicine 

at Mount Sinai, New York, NY; 2 Liver Diseases, Icahn School 

of Medicine at Mount Sinai, New York, NY; 3 Pathology, Icahn 

School of Medicine at Mount Sinai, New York, NY; 4 Surgery, 

Recanati/Miller Transplantation Institute; Icahn School of Medicine 

at Mount Sinai, New York, NY 

Background: The Milan criteria are widely applied in selecting 

and listing patients with hepatocellular carcinoma (HCC) for 

liver transplantation (LT). They fundamentally depend on the 

sensitivity of radiologic imaging in detecting lesions. We aimed 

to compare radiologic detection with pathologic assessment of 

HCC in explanted livers as well as to identify clinical features 

that might impact this correlation. Design: At our institution, LT 

candidates undergo CT scan or MRI every 3-6 months depending 

on if they have HCC and if it is being actively treated. 

Per protocol, all explanted livers are weighed, sectioned at 

0.5cm intervals, and reviewed by the pathologist. For the current 

study, retrospective clinical data were gathered, including 

demographics, MELD score, # of lesions seen on imaging, and 

history of locoregional therapy (LRT). Retrospective histopathological 

review of liver explant sections was performed by a 

liver pathologist (MIF) blinded to patient identities. The degree 

of cirrhosis based on the Laennec sub-classification (4A=incomplete 

cirrhosis, 4B=nodules with thin fibrous septa, 4C=nodules 

surrounded by thick fibrous septa) was assessed. Correlation of 

the number of tumors found on radiologic imaging and pathologic 

examination was performed. Results: 148 patients with 

HCC who underwent LT over a 36-month period were studied. 

There were 114M & 34F, 60.2±7.2 years of age. 105 had 

LT for HCV, 17 for NASH, 19 for HBV, and the rest for other 

etiologies. 66 patients (45%, Group A) showed concordance 

between imaging and pathologic findings of HCC whereas 82 

(55%, Group B) did not. Laennec staging was performed on 

patients over the first 21-month period. 32/34 in Group A and 

26/27 in Group B had cirrhosis. More Group B than Group 

A patients had class 4C cirrhosis (p=0.028). Patients with a 

lack of concordance also had a higher creatinine (p=0.019) 

and MELD score (p=0.021). There was a trend towards smaller 

liver volumes in this group as well. No significant difference 

was found in age, sex, time between LT and last imaging 

study, and LRT. 39 (48%) in Group B had tumors ≥1cm not 

detected by pre-LT imaging. Conclusion: There was a high rate 

of non-correlation between radiologic and pathologic findings 

in our cohort of patients with known HCC. We found that 

HCC is detected less well on imaging in patients having more 

advanced fibrosis, as well as in patients with renal dysfunction 

and higher MELD scores. Current radiologic modalities may 

not be sensitive enough to identify HCC lesions in this population 

and thus underestimate tumor burden at time of LT. This 

may be due to less optimal penetration of radiologic contrast 

into their liver parenchyma. 

871 

Using Biomarkers to Predict Progression to End-stage 

Renal Disease within 6 Months of Liver Transplant 

Joseph Bahng, Peter L. Abt, Deirdre Sawinski, Anirban Ganguli, 

Debra McCorriston, Mary Ann Lim, Kimberly A. Forde; Perelman 

School of Medicine at the University of Pennsylvania, Philadelphia, 

PA 

Background Liver transplant (LT) recipients are at increased 

risk for chronic kidney disease and end-stage renal disease 

(ESRD). Creatinine, the standard for assessment of renal function, 

provides limited prognostic information about the likelihood 

of recovery from acute kidney injury (AKI). Our aim was 

to create a predictive model for progression to ESRD within 6 

months of LT that incorporates clinical data and biomarkers. 

Methods We enrolled 202 patients listed for LT at our center 

from 2012-2014. Serum and urine samples were collected at 

enrollment, at LT, and 1 year thereafter. Clinical and demographic 

data were also collected. Patients were followed until 

they developed the primary outcome, ESRD, defined as glomerular 

filtration rate


872 

Traveling for a Liver Transplant in the US does not 

Result in a Lower Match MELD for the Majority of 

Patients 

Catherine T. Frenette 1 , Ann M. Harper 2 , Angeles Baquerizo 1 , Randolph 

L. Schaffer 1 , Jonathan S. Fisher 1 , Christopher L. Marsh 1 ; 

1 Organ Transplantation, Scripps Clinic, La Jolla, CA; 2 United Network 

for Organ Sharing, Richmond, VA 

Aim: Given the current geographic disparities in the Unites 

States in liver allocation for orthotopic liver transplantation 

(OLT), there is a segment of the population that may travel 

to a different donor service area other than their home. The 

purpose of this study was to determine if patients who traveled 

for OLT were more likely to be transplanted at a lower MELD. 

Methods: The OPTN database was queried for all patients who 

underwent a deceased donor liver transplant from 2010-2014. 

Patients with unknown home zip code were excluded (N=340). 

Demographic, insurance, transplant, exception status and survival 

data were analyzed (N=30,436). Patients who traveled 

outside of their home area (Tr) were compared to patients 

who underwent OLT at home (NTr). Results: From 2010-2014, 

25.6% of patients traveled outside of their home donor service 

area (N=7780). Of these patients, 6925 (89%) traveled outside 

of their home state. The mean distance traveled was 368 

miles for Tr vs 50.3 miles for NTr. There was no difference in 

gender between the groups (men 66.9% Tr vs 65.1% NTr). 

There was a higher proportion of pediatric patients in the Tr 

group (10.2% vs 6.9%, p


874 

Coronary Angiography in Patients being evaluated for 

Orthotopic Liver Transplantation 

Feng Li, A. James Hanje, Khalid Mumtaz, Robert B. Kirkpatrick, 

Douglas M. Levin, Elmahdi Elkhammas, Sylvester Black, Alice Hinton, 

Anthony Michaels; The Ohio State University Medical Center, 

Columbus, OH 

Purpose: The ideal preoperative cardiac screening study in 

patients with end-stage liver disease undergoing an orthotopic 

liver transplantation (OLT) evaluation has yet to be determined. 

Left heart catheterization (LHC) can be considered for cardiac 

screening in patients with coronary artery disease (CAD) risk 

factors. We aim to identify the patients that would most benefit 

from a LHC during an OLT evaluation. Methods: We retrospectively 

identified patients who underwent a LHC during an 

OLT evaluation at our center between May 2009 and February 

2015. Demographic, clinical, laboratory, and procedural 

data were collected and analyzed. Patients considered for LHC 

included patients with abnormal non-invasive testing, symptoms 

consistent with CAD, evidence of structural abnormality 

in a technically sufficient study, or associated risk factors such 

as tobacco use, diabetes mellitus, family history of premature 

CAD, age (> 45 men and >55 women), hyperlipidemia, hypertension, 

left ventricular hypertrophy, obesity (BMI > 35), and 

non alcoholic fatty liver disease (NASH) or alcoholic liver disease. 

Results: 126 patients were included in the study. 25% 

had a negative LHC, 64% had a positive LHC without intervention, 

and 11% had a positive LHC with non-surgical intervention. 

82% of patients with NASH had a positive LHC and 

14% required non-surgical intervention; 62% of patients with 

alcoholic liver disease had a positive LHC and 8% required 

non-surgical intervention; 77% of patients with hepatitis C had 

a positive LHC and 17% required non-surgical intervention. Of 

the 75% of patients with a positive LHC, 25% underwent OLT 

at our medical center, and 21% of the patients were deferred 

for transplant listing at our center with the LHC findings contributing 

to their deferment. Age (p=0.009) and a history of 

hyperlipidemia (p=0.025) were the only two statistically significant 

risk factors in predicting a positive coronary angiography 

on univariate analysis. A forward stepwise process was used 

to build a model containing significant predictors of a positive 

coronary angiography. Age was a significant predictor 

(p=0.023) and was associated with higher odds of a positive 

coronary angiography with Odds Ratios of 1.07 (95% Confidence 

Interval (CI), 1.01-1.13), 1.40 (95% CI, 1.05-1.87) and 

1.95 (95% CI, 1.10-3.48) for a 1 year, 5 year, and 10 year 

age increase, respectively. Conclusion: Coronary Angiography 

is an appropriate cardiac screening tool in a select group of 

patients undergoing an OLT evaluation particularly in patients 

with increasing age and history of hyperlipidemia. 

Disclosures: 


Anthony Michaels - Advisory Committees or Review Panels: Gilead, BMS, Janssen; 


The following authors have nothing to disclose: Feng Li, Khalid Mumtaz, Robert 

B. Kirkpatrick, Douglas M. Levin, Elmahdi Elkhammas, Sylvester Black, Alice 

Hinton 

875 

Long Term Survival Of Liver Grafts From Elderly Donors 

Marius Braun 1 , Eviatar Nesher 2 , Michal Cohen-Naftaly 1 , Assaf 

Issachar 1 , Amir Shlomai 1 , Yael Harif 1 , Sigal Eisner 2 , Ran Tur- 

Kaspa 1 , Eytan Mor 1 ; 1 Liver institute, Rabin Medical Center, Petach 

tiqva, Israel; 2 Transplant department, Rabin Medical Center, 

Petach tiqva, Israel 

Introduction ; There is an increasing gap between the supply 

of organ donations and need for liver grafts. The decreasing 

quality of the grafts due to aging and overweight population 

is expected to lead to a decrease in transplant volume 

from deceased donors. The use of extended criteria for grafts 

including aged donors may expand the dwindling donor 

pool. Traditionally, older donors have been regarded as highrisk 

transplants, but recently encouraging results have been 

reported. We analyzed our own experience in recipients 

of grafts from donors older than 70 years. Methods : Retrospective 

analysis of a prospectively maintained transplantation 

data base. Recipients of livers older than 70 years were 

compared to recipients from donors younger than 60 y and 

donors between 61 and 69 y with regard to long term survival 

and complications. Statistical analysis included Kaplan Meyer 

survival curves, chi squre and regression analysis. Results : 

42 patients (25 males) received grafts from donors older than 

70 years. Their characteristics and outcomes were compared 

with 238 patients who received grafts from donors less than 

60 years of age and with 60 patients with grafts from donors 

between 61 and 69 years of age. In the group of donors over 

70, the mean donor age was 74.3 y (SD 2.8 ) and the recipients 

age was 57.8y (range 22-74, SD 9.4).The indications for 

transplantation varied: eleven cases of HBV cirrhosis, five HCV 

cirrhosis and one hepatocellular carcinoma. Mean MELD at 

transplant was 20.8. The long term patient survival at 5 years 

was 70% and was similar among all the age groups. Conclusions: 

Good quality grafts from elderly donors maybe used 

to extend the donor pool. New techniques might improve the 

preservation and assessment of these grafts to decrease shortterm 

complications and mortality 

Disclosures: 

The following authors have nothing to disclose: Marius Braun, Eviatar Nesher, 

Michal Cohen-Naftaly, Assaf Issachar, Amir Shlomai, Yael Harif, Sigal Eisner, 

Ran Tur-Kaspa, Eytan Mor 

876 

Controlled attenuation parameter (CAP) assessment may 

be clinically useful to screen hepatic steatosis for liver 

transplant purposes 

Youngmi Hong 1 , Ki Tae Yoon 1 , Mong Cho 1 , Daehwan Kang 1 , 

Hyungwook Kim 1 , Cheol Woong Choi 1 , SuBum Park 1 , Jeong 

Heo 2 , Hyun Young Woo 2 , Won Lim 2 ; 1 Pusan National University 

Yangsan Hospital, Yangsan, Korea (the Republic of); 2 Pusan 

National University Hospital, Busan, Korea (the Republic of) 

Backgroud & Aims: The presence of hepatic steatosis is associated 

with an increased risk of graft loss. The controlled 

attenuation parameter (CAP) using transient elastogarphy is a 

noninvasive method of assessing hepatic steatosis. However, 

the data on the diagnosis performance of CAP is still limited. 

Therefore, we assessed the accuracy and the efficacy of CAP 

for the detection of hepatic steatosis in potential liver donors. 

Methods: We enrolled potential liver donors and all patients 

underwent CAP assessment, MRI (Dixon in phase/out of phase 

(Dixon IP/OP) with/without fat saturation images) and ultrasonography-guided 

liver biopsy. Results: A total of 30 potential 

liver donors were included: 24 were male and median 

age was 27 years. Clinical and laboratory variables were


analyzed according to CAP values. The median CAP value 

was 246.4 dB/m (range, 129-371) and CAP value was positively 

correlated with alanine aminotransferase (ALT) (r=0.516, 

P


879 

Ethanol-Mediated Lipocalin 2 Induction Selectively 

Impairs Hepatic Chaperone-Mediated Autophagy and 

Causes Alcoholic Steatosis in Mice 

Xudong Hu 1,2 , Jiayou Wang 1,3 , Alvin Jogasuria 1 , Kwangwon Lee 1 , 

Jiashin Wu 1 , Min You 1 ; 1 Pharmaceutical Sciences, Northeast Ohio 

Medical University (NEOMED), Rootstown, OH; 2 Department of 

Biology, Shanghai University of Traditional Chinese Medicine, 

Shanghai, China; 3 Department of Anatomy, Guangzhou University 

of Chinese Medicine, Guangzhou, China 

Lipocalin-2 (LCN2) [also named as SIP24/24 p 

3 in mouse and 

neutrophil gelatinase-associated lipocalin (NGAL) in human] 

has been recently implicated as a critical player in alcohol-induced 

fatty liver disease (AFLD). However, the cellular and 

molecular mechanisms for its role in mediating detrimental 

action of ethanol in the liver are poorly understood. We investigated 

the in vivo function of LCN2 in the development of AFLD 

by pair-feeding wild-type (WT) and LCN2 knockout (Lcn2KO) 

mice using Lieber-DeCarli ethanol-containing diets for 4 weeks. 

In WT mice, chronic ethanol administration significantly inhibited 

the mRNA expression of hepatic chaperone-mediated autophagy 

activity with decreased heat shock protein (Hsp) a8 

(Hspa8), Hsp90aa1 and hsp70 compared to pair-fed controls. 

Remarkably, mRNA levels of Hspa8, Hsp90aa1 or hsp70 in 

the livers of chronically ethanol-fed Lcn2KO mice were completely 

restored to the pair-fed control levels. Consistently, the 

mRNA levels of lysosome-associated membrane protein type 

2A (LAMP-2A) and co-chaperone BAG2, two molecules also 

known to participate in chaperone-mediated autophagy, were 

significantly inhibited in ethanol-fed WT mice but partially 

and significantly restored in the livers of ethanol-fed Lcn2KO 

mice. Those results clearly demonstrate that LCN2 deficiency 

selectively stabilizes and activates hepatic chaperone-mediated 

autophagy in chronically ethanol-fed Lcn2KO mice. The 

autophagic flux status was further estimated by quantitating the 

protein levels of p62, an autophagy-selective substrate. Compared 

to WT control mice, chronic ethanol feeding substantially 

inhibited p62 protein levels, and LCN2 deficiency suppressed 

p62 to the similar extent. Interestingly, chronic ethanol administration 

to Lcn2KO mice augmented inhibition of p62 protein 

expression compared to all other groups. Consistent with these 

in vivo findings, the mRNAs of Hspa8, Hsp90aa1 and hsp70 

were significantly reduced by treatment of recombinant LCN2 

in a dose-dependent manner in mouse AML-12 hepatocytes. 

Collectively, our studies demonstrate that ethanol exposure 

selectively inhibits hepatic chaperone-mediated autophagy 

and disrupts autophagic flux via LCN2 induction in mice. Ethanol-mediated 

disruption of hepatic chaperone-mediated autophagy 

and autophagic flux via LCN2 contributes, at least in 

part, to the development of alcoholic fatty liver in mice. 

Disclosures: 

The following authors have nothing to disclose: Xudong Hu, Jiayou Wang, Alvin 

Jogasuria, Kwangwon Lee, Jiashin Wu, Min You 

Disclosures: 

The following authors have nothing to disclose: Kyota Fukazawa, Alexander A. 

Vitin, Kenneth Martay, Jorge Reyes 

880 

Hepatocyte-specific depletion of UBXD8 accelerates 

fibrosis in a mouse non-alcoholic steatohepatitis model 

Norihiro Imai 1,2 , Yoji Ishizu 1 , Teiji Kuzuya 1 , Takashi Honda 1 , 

Kazuhiko Hayashi 1 , Masatoshi Ishigami 1 , Yoshiki Hirooka 1 , Toyoshi 

Fujimoto 2 , Hidemi Goto 1 ; 1 Gastroenterology and Hepatology, 

Nagoya University Graduate School of Medicine, Nagoya, Japan; 

2 Anatomy and Molecular Cell Biology, Nagoya University Graduate 


[Background] We previously reported that UBXD8 in lipid 

droplets is a key molecule for the degradation of lipidated 

ApoB in hepatocytes. More recently, we found that hepatocyte-specific 

UBXD8-deficient mice (U8-LKO) fed a high-fat 

diet develop periportal macrovesicular steatosis accompanied 

by a decrease in VLDL secretion (Imai et al, PLOS ONE 10, 

e0127114, 2015). These results demonstrated that UBXD8 is 

a crucial factor to regulate VLDL secretion from the liver and 

suggested that dysfunction of UBXD8 may aggravate or even 

cause NAFLD/NASH. To address this possibility, we studied 

U8-LKO mice fed a NASH model diet. [Method] U8-LKO mice 

and their age-matched littermates (control) were fed with three 

different diets: (1) normal chow diet (CLEA Rodent Diet CE-2), 

(2) high-fat diet (CLEA Rodent Diet Quick Fat; 14.4% fat), and 

(3) choline-deficient high-fat diet (CDHF, Oriental Yeast; 60% 

fat). Blood samples were analyzed biochemically. Paraffin sections 

were stained with hematoxylin and eosin or Sirius red to 

evaluate morphological changes. [Results] (1) With a normal 

diet, liver sections did not exhibit any morphological difference 

between U8-LKO and control mice. No sign of hepatic fibrosis 

or steatosis was observed. (2) When mice were fed a high-fat 

diet for 30 weeks, U8-LKO mice showed periportal steatosis, 

as we reported previously. Even when steatosis was prominent, 

no sign of fibrosis was observed in control or U8-LKO mice. 

(3) When mice were fed a CDHF diet for two weeks, serum 

AST and ALT levels were greater in U8-LKO mice than in control 

mice (AST, 331 IU/l vs. 227 IU/l; ALT, 334 IU/l vs. 238 

IU/l). Serum bilirubin and ALP levels were markedly higher 

in U8-LKO mice than in control mice (bilirubin, 96.7 μg/dl 

vs. 47.5 μg/dl, p = 0.036; ALP, 840 IU/l vs. 288 IU/L, p = 

0.023), although food consumption and body weight changes 

were similar in both groups. Under these conditions, the liver 

of U8-LKO mice had a lumpy surface, and marked periportal 

and perisinusoidal fibrosis was observed histologically. Control 

mice showed no fibrosis of the liver. The Sirius red-positive 

area was also significantly greater in U8-LKO mice than in 

control mice (4.67% vs. 0.75%, p = 0.004). [Conclusion] With 

only two weeks of CDHF diet feeding, U8-LKO mice exhibited 

remarkable fibrosis; this was not observed with a normal or 

simple high-fat diet. These results indicate that UBXD8 functionality 

can be largely compensated for in the normal dietary 

condition or even with mild fat loading, but that UBXD8 plays 

a crucial role when the liver is exposed to intense dietary challenge. 

It would be worthwhile to determine if UBXD8 functions 

similarly in the human liver. 

Disclosures: 



The following authors have nothing to disclose: Norihiro Imai, Yoji Ishizu, 

Teiji Kuzuya, Takashi Honda, Kazuhiko Hayashi, Masatoshi Ishigami, Yoshiki 

Hirooka, Toyoshi Fujimoto


881 

Regulation of Adaptive Thermogenesis by the Gut 

Microbiome: Implications for Non-Alcoholic Fatty Liver 

Disease (NAFLD) Pathogenesis 

Tibor I. Krisko 1,2 , Hayley T. Nicholls 1 , Katherine B. Leclair 1 , Alexander 

S. Banks 1 , David E. Cohen 1 ; 1 Medicine, Brigham and Women’s 

Hospital, Boston, MA; 2 Medicine, VA Medical Center, Boston, 

MA 

Background: The gut microbiome plays a critical role in the 

pathophysiology of both obesity and non-alcoholic fatty liver 

disease (NAFLD). While an altered energy balance is known 

to be associated with liver injury, the mechanisms by which the 

gut microbiome controls energy homeostasis and thus may contribute 

to NAFLD are incompletely understood. Recent studies 

have shown that increasing thermogenesis ameliorates hepatic 

steatosis via both direct and indirect mechanisms (Wang, Nat 

Med 2014;20:1436). Aim: This study was designed to elucidate 

the contribution of the gut microbiome to adaptive thermogenesis, 

as reflected by alterations in energy expenditure 

under conditions of varied thermal stress. Methods: We utilized 

a temperature-controlled comprehensive lab animal monitoring 

system that was custom-designed to maintain sterile conditions. 

Rates of O 2 

consumption (VO 2 

) and CO 2 

production (VCO 2 

) 

were quantified by indirect calorimetry in control and germ-free 

C57BL/6J mice as functions of variations in ambient temperatures 

that ranged from thermoneutrality (30 °C) to severe cold 

stress (4 °C). Energy expenditure (kcal/72 h) was estimated 

according to the formula VO 2 

(3.82 + 1.22RER) and respiratory 

exchange ratio (RER) was calculated as (VCO 2 

/VO 2 

). 

Physical activity was recorded as cumulative beam breaks per 

hour over 72 h. Body composition was determined by NMR 

spectroscopy and blood glucose concentrations using a glucometer. 

Germ-free status was confirmed at the beginning and 

end of experiments by quantitative culture and gram staining of 

stool. Results: Compared to controls, germ-free mice exhibited 

lower energy expenditure, both at thermoneutrality (18.9 ± 0.2 

vs 21.1 ± 0.3, p < 0.01) and in the cold (54.7 ± 0.5 vs 58.2 

± 0.2, p < 0.05). This occurred despite increased physical 

activity in germ-free mice that was most pronounced at 4 °C 

(1023 ± 54 vs 1542 ±127, p < 0.05). Germ-free mice exhibited 

a greater percentage of adipose tissue mass compared 

to controls. Plasma glucose concentrations were reduced and 

values of RER were lower in germ-free mice at 4 °C, indicating 

increased lipid utilization. Conclusions: Reductions in energy 

expenditure and increased adiposity despite increased physical 

activity support a key role for the gut microbiome in promoting 

adaptive thermogenesis across a broad range of ambient 

temperatures. In addition, reduced RER values and plasma 

glucose concentrations in germ-free mice indicate that the gut 

microbiome promotes the utilization of dietary carbohydrate as 

an energy substrate. We speculate that impaired thermogenesis 

in the setting of dysbiosis contributes to the pathogenesis 

of NAFLD. 

Disclosures: 



The following authors have nothing to disclose: Tibor I. Krisko, Hayley T. Nicholls, 

Katherine B. Leclair, Alexander S. Banks 

882 

The dual FXR/TGR5 agonist INT-767 counteracts nonalcoholic 

steatohepatitis in a rabbit model of high fat 

diet-induced metabolic syndrome 

Linda Vignozzi 1 , Ilaria Cellai 1 , Sandra Filippi 4 , Paolo Comeglio 1 , 

Erica Sarchielli 2 , Annamaria Morelli 2 , Elena Maneschi 1 , Gabriella 

Barbara Vannelli 2 , Luciano Adorini 3 , Mario Maggi 1 ; 1 Dep. 

of Experimental and Clinical Biomedical Sciences, University of 

Florence, Florence, Italy; 2 Dep. of Experimental and Clinical Medicine, 

University of Florence, Florence, Italy; 3 Intercept Pharmaceuticals, 

New York, NY 10011, NY; 4 Interdepartmental Laboratory 

of Functional and Cellular Pharmacology of Reproduction, Dep. of 

NEUROFARBA and Dep. of Experimental and Clinical Biomedical 

Sciences, University of Florence, Florence, Italy 

Farnesoid X receptor (FXR) and Takeda G protein-coupled 

receptor 5 (TGR5) are interesting pharmacological targets for 

the treatment of liver and metabolic diseases. FXR-deficient 

mice on a high-fat diet (HFD) exhibit massive hepatic steatosis, 

necro-inflammation and fibrogenesis. Moreover, pharmacological 

activation of TGR5 in mice promotes protective mechanisms 

in biliary epithelial cells, inhibits hepatic and systemic 

inflammation. Thus, we hypothesized that a FXR/TGR5 dual 

agonist would ameliorate features of nonalcoholic steatohepatitis 

(NASH) in a rabbit model of metabolic syndrome (MetS). 

Treatment with increasing doses of the dual FXR/TGR5 agonist 

INT-767 (3,10,30mg/Kg/day, 5 days a week for 12 weeks) 

in a rabbit model of HFD-induced MetS, characterized also 

by NASH, dose-dependently reduced several MetS-associated 

alterations, including hepatomegaly, insulin resistance, 

increase of ALT, glucose and cholesterol levels, while significantly 

increasing HDL levels. ALT was positively associated 

with all MetS parameters; however introducing all MetS factors 

in a multivariate analysis, only total cholesterol levels resulted 

positively associated with ALT level (Adj.r: 0.493, p=0.014). 

High macrophage M1pro-inflammatory/M2 anti-inflammatory 

ratio was observed in MetS-induced NASH, which was 

independently associated with serum ALT levels (Adj.r: 0.322, 

p=0.032). HFD-induced increase in M1/M2 ratio was reduced 

by INT-767 treatment and M2 macrophage markers (IL-10,T- 

GFβ) were increased. Genes related to neutrophil apoptosis/ 

apoptotic-neutrophil clearance (lactoferrin, eNOS, RAGE) 

and to extracellular matrix degradation (MMP2, TIMP2) were 

also increased by INT-767 treatment. INT-767 also reduced 

liver expression of IL-6, which preferentially skews the Th cell 

response towards a Th17-phenotype, while increasing Foxp3 

expression, a Treg cell marker. Thus these data indicate that 

INT-767 can promote the neutrophil and macrophage-driven 

resolution phase of inflammation and fibrosis regression. In 

addition, INT-767 increased genes related to hepatic fatty acid 

metabolism (PPARa, AR, CD36) and lipid droplet formation 

(SNAP23, VAMP4,syntaxin5, perilipin) therefore suggesting 

that INT-767 counteracts excess fatty acid mediated lipotoxicity 

in the liver. Genes related to insulin signaling (IRS1, SREBP1, 

G6Pase, and PEPCK) were also increased by INT-767. Finally, 

immunohistochemical studies demonstrated that INT-767 treatment 

significantly reduced both HFD-induced liver inflammation 

and fibrosis. In conclusion, INT-767 treatment counteracts 

NASH in a rabbit model of HFD-induced MetS by promoting 

insulin sensitivity, resolution of inflammation and fibrosis regression. 

Disclosures: 


Mario Maggi - Consulting: BAYER, ELI LILLY, MENARINI, PROSTRAKAN, INTER- 

CEPT 

The following authors have nothing to disclose: Linda Vignozzi, Ilaria Cellai, 

Sandra Filippi, Paolo Comeglio, Erica Sarchielli, Annamaria Morelli, Elena Maneschi, 

Gabriella Barbara Vannelli


883 

Lipid-induced Endoplasmic Reticulum Stress But Not 

Hepatic Steatosis Contributes to Blockage of Autophagosome-Lysosome 

Fusion 

Koichiro Miyagawa, Shinji Oe, Yuichi Honma, Masaru Harada; 

University of Occupational and Environmental Health, Kitakyushu, 

Japan 

Purpose: Autophagy is crucial for intracellular quality control of 

proteins and intracellular organelles in various tissues. Several 

studies showed that blockage of hepatic autophagic degradation 

system occurred in obese, and that had an important role 

in the development of nonalcoholic fatty liver disease (NAFLD). 

However, the mechanism of this blockage has not been fully 

elucidated. In this study, we evaluated how lipid overload is 

linked to impairment of autophagy in hepatocytes. Methods: 

We investigated the effect of lipid overload on hepatic autophagy 

in cultured hepatocytes-derived cells treated with monounsaturated 

fatty acids (MUFAs) or saturated fatty acids (SFAs). 

Autophagic flux was analyzed by immunoblotting and fluorescence 

microscopy. We also evaluated the relationship between 

hepatic steatosis, endoplasmic reticulum (ER) stress, and autophagy 

in this model. Results: SFAs but not MUFAs induced ER 

stress and accumulation of autophagosomes despite of lower 

accumulation of lipid droplets compared with MUFAs. Microtubule-associated 

protein 1 light chain 3 (LC3) turnover assay 

demonstrated that SFAs induced the reduction of the rate of 

lysosomal degradation of autophagosomes without change 

in autophagosome synthesis. Moreover, we monitored autophagic 

flux in cells transfected with mRFP-GFP tandem fluorescence-tagged 

LC3 (tf-LC3). GFP fluorescence is quenched in 

the lysosome but mRFP is not, indicating that GFP and mRFP 

are co-localized before the fusion with lysosomes. SFAs but not 

MUFAs induced GFP and mRFP overlapping, indicating that 

SFAs induced accumulation of autophagosomes. SFAs also 

suppressed co-localization of mRFP and lysosome-associated 

membrane protein 1 (Lamp1), suggesting that SFAs-induced 

impairment of autophagic flux was due to blockage of autophagosome-lysosome 

fusion. Furthermore, we assessed lysosomal 

acidification. Cells treated with SFA were labeled for 

Lysotracker Red and then stained with Lamp1. We could not 

identify Lysotracker-negative Lamp1 in SFAs-treated cells as 

well as vehicle, suggesting that lysosomal acidification was not 

impaired by SFAs treatment. We also revealed that SFAs did 

not affect cathepsin activity. These results suggested that SFAs 

impaired autophagosome-lysosome fusion without change in 

lysosomal function. Moreover, we found that this impairment 

occurred in an ER stress-dependent manner. Conclusions: The 

disturbance of autophagic flux in NAFLD is due to the blockage 

of autophagosome-lysosome fusion, and that involves in the 

degree of ER stress but not the intracellular accumulation of 

lipid droplets. 

Disclosures: 

The following authors have nothing to disclose: Koichiro Miyagawa, Shinji Oe, 

Yuichi Honma, Masaru Harada 

884 

CHOP and the Unfolded Protein Response Regulate 

NF-κB Activity through IRAK2 in the Pathogenesis of 

Non-Alcoholic Steatohepatitis 

Jeffrey A. Willy 1 , James L. Stevens 1 , Howard C. Masuoka 2 , Ronald 

C. Wek 1 ; 1 Biochemistry and Molecular Biology, Indiana University 

School of Medicine, Indianapolis, IN; 2 Department of Medicine, 

Indiana University School of Medicine, Indianapolis, IN 

Free fatty acid (FFA) induction of cell death is an important feature 

of NASH and has been associated with disruption of the 

endoplasmic reticulum and activation of the Unfolded Protein 

Response (UPR). The mechanisms by which this stress pathway 

results in deleterious effects at the organ and cellular levels 

are not well understood. In this study, we examined the role of 

the UPR in NASH. The UPR features transcriptional and translational 

control of gene expression involved in stress remediation, 

including the transcription factor CHOP, which can trigger 

death processes during chronic endoplasmic reticulum stress. 

Primary hepatocytes and HepG2 cells were exposed to physiologic 

levels of either saturated or unsaturated FFA. We found 

that the saturated FFA palmitate, but not the unsaturated FFA 

oleate, induced hepatocyte cell death as a result of inhibition 

of autophagic flux in a CHOP-dependent manner. Additionally, 

saturated FFA exposure resulted in activation of NF-κB through 

expression of IRAK2, resulting in secretion of many cytokines, 

including TNFα and IL-8, which contribute to both hepatic cell 

death and inflammation. Depletion of CHOP or the RelA subunit 

of NF-κB in hepatocytes by shRNA alleviated both autophagy 

and cytokine secretion resulting in enhanced cell viability 

and lowered inflammatory responses during exposure to saturated 

FFA. We next investigated the relevance of these findings 

in human liver disease. We showed that the CHOP-IRAK2- 

NF-κB pathway is upregulated in liver biopsies from patients 

with NASH, in combination with inhibition of autophagic flux, 

as measured by accumulation of LC3b and P62. Secretion of 

CHOP-dependent cytokines such as TNFα and IL-8 are also 

increased in the sera of NASH patients. Interestingly, while 

UPR activation was conserved in primary rodent hepatocytes, 

there was no induction of IRAK2 and NF-κB in primary rodent 

hepatocytes following saturated FFA exposure, providing a 

potential explanation for the shortcomings in regards to inflammation 

and fibrosis in current rodent models of NASH relative 

to the human disease. In conclusion, we have identified a novel 

pathway that is upregulated in NASH patients that plays an 

active role in both the hepatotoxicity and inflammation at the 

level of the hepatocyte following excess saturated FFA. A better 

understanding of the mechanism by which the UPR contributes 

to hepatocyte toxicity during FFA exposure may provide new 

approaches for the diagnosis and treatment of liver disease. 

Disclosures: 

The following authors have nothing to disclose: Jeffrey A. Willy, James L. Stevens, 

Howard C. Masuoka, Ronald C. Wek 

885 

Deletion of G-protein-coupled bile acid receptor (Gpbar- 

1, Tgr5) alleviates fasting induced hepatic steatosis by 

altering hepatic lipid metabolism 

Ajay C. Donepudi, Shannon M. Boehme, John Chiang; Northeast 

Ohio Medical University, Rootstown, OH 

Introduction: Bile acids and its receptors such as farnesoid X 

receptor (Fxr) and G-protein-coupled bile acid receptor (Gpbar- 

1, Tgr5) play a major role in nutrient homeostasis. Tgr5 has 

been shown to regulate glucose homeostasis by regulating insulin 

secretion. Tgr5 is also involved in regulating basal metabolic


rates by regulating endocrine hormone levels such as thyroid 

hormone levels. Fasting is an adaptive mechanism developed 

in mammals for nutrient depravation. Fasting is characterized 

with increase in free fatty acid levels in serum and liver causing 

hepatic steatosis along with several other physiological 

changes. These physiological changes in fatty acid metabolism 

during fasting are similar to diabetes condition. In this study 

we aim to identify role of Tgr5 in fatty acid metabolism using 

fasting model. Methods: Male C57BL/6J (WT) and Tgr5 knock 

out (Tgr5-/-) mice in pure C57BL/6J background were either 

fed ad libitum or fasted for 24 hrs. Lipid profiles of serum, liver 

and adipose tissues were analyzed. Quantitative real-time PCR 

was used to assay mRNA expression levels of the genes in bile 

acid, cholesterol and lipid metabolism. A Tgr5 specific agonist 

INT-777 (Intercept Pharma) was used to activate Tgr5 signaling 

in mice. Results: Fasting increased hepatic lipid accumulation 

in both WT and Tgr5-/- mice. Lack of Tgr5 attenuated fasting-induced 

hepatic triglyceride and fatty acids accumulation compared 

to WT mice, without changes in adipose tissue lipolysis. 

Gene expression analysis showed Tgr5-/- mice have increased 

expression of hepatic lipolysis genes such as Cpt1, Pgc-1α 

and autophagy gene LC3II levels. Comprehensive laboratory 

animal monitoring system (CLAMS) metabolic studies showed 

Tgr5-/- mice have increased physical activity and energy 

expenditure. Tgr5-/- mice also showed decrease in expression 

of hepatic fatty acid transporter Cd36 and decreased hepatic 

fatty acid uptake. Additionally, Tgr5-/- mice had increased 

hepatic Stat5 activation, which is known to regulate hepatic 

lipid metabolism. Moreover, a Tgr5 agonist INT-777 treatment 

increased hepatic Cd36 expression and decreased hepatic 

Lc3II levels in fasted mice. Conclusion: Although Tgr5 is not 

expressed in hepatocytes, Tgr5 may play a role in regulation 

of hepatic fatty acid metabolism during fasting. Lack of 

Tgr5 attenuates fasting-induced hepatic steatosis by decreasing 

hepatic fatty acid uptake and increasing hepatic lipolysis. 

Disclosures: 

The following authors have nothing to disclose: Ajay C. Donepudi, Shannon M. 

Boehme, John Chiang 

886 

Hepatic NOD-like receptors, pyrin domain-containing 

3 (NLRP3) inflammasome activation is associated with 

histological severity in patients with nonalcoholic fatty 

liver disease 

Hironori Mitsuyoshi, Kohichiroh Yasui, Tasuku Hara, Hiroyoshi 

Taketani, Hiroshi Ishiba, Akira Okajima, Yuya Seko, Atsushi 

Umemura, Taichiro Nishikawa, Kanji Yamaguchi, Michihisa Moriguchi, 

Yoshio Sumida, Masahito Minami, Yoshito Itoh; Kyoto Prefectural 

University of Medicine, Kyoto, Japan 

Background and Aims: Nonalcoholic fatty liver disease 

(NAFLD) is the hepatic manifestation of metabolic syndrome, 

which is associated with dysregulated inflammasome activation 

caused by obesity. Inflammasomes are intracellular multiprotein 

complexes, comprising nucleotide-oligomerization and binding 

domain (NOD)-like receptors, the apoptosis-associated specklike 

protein containing a CARD (ASC), and procaspase-1. 

Assembly of these components in response to metabolic stress 

results in caspase-1 activation, leading to interleukin-1 beta 

(IL-1β) and IL-18 activation, which exacerbates inflammation 

in systemic organs, including the liver. Here, we examined the 

role of inflammasomes in the development of NAFLD. Methods: 

Biopsy-proven patients with NAFLD (n = 91) were enrolled. 

Liver histology was assessed according to the NAFLD activity 

score (NAS). Hepatic (n = 91) and blood (n = 37) levels of the 

NOD-like receptor family, pyrin domain-containing 3 (NLRP3), 

ASC, procaspase-1, IL-1β, and IL-18 were quantified by realtime 

polymerase chain reaction (PCR). Adiponutrin polymorphisms 

(rs738409, C>G) were determined by TaqMan PCR. 

Serum IL-1β and IL-18 levels were measured by ELISA and 

liver tissue caspase-1 expression was evaluated by immunostaining. 

Results: Hepatic mRNA levels of NLRP3, ASC, procaspase-1, 

IL-1β, and IL-18 were significantly higher in patients 

with NAFLD compared with control livers. Blood procaspase-1 

mRNA levels were significantly higher in NAFLD patients compared 

with healthy controls. In contrast to the blood mRNA 

levels, hepatic mRNA levels of NLRP3 inflammasome components 

were significantly associated with adiponutrin G alleles. 

Hepatic procaspase-1 and IL-1β mRNA levels correlated significantly 

with lobular inflammation, hepatocyte ballooning, 

and total NAS. Serum IL-18 levels were significantly higher 

in NAFLD patients compared with controls, while IL-1β levels 

exhibited a non-significant increase. Serum IL-1β and IL-18 levels 

were significantly correlated with steatosis, total NAS and 

serum transaminase levels. Caspase-1-positive cells were scattered 

within the portal tracts, inflammatory foci, and ballooning 

hepatocytes. Immunofluorescence staining showed colocalization 

of caspase-1 with the macrophage marker CD68. Conclusions: 

Our results suggest that NLRP3 inflammasomes are 

primed in the liver in NAFLD patients and that this process is 

influenced by adiponutrin genotypes. Furthermore, our results 

indicate that NLRP3 inflammasomes are activated exclusively 

in Kupffer cells or infiltrating macrophages, leading to histological 

NAFLD progression through IL-1β and IL-18 production. 

Disclosures: 

Yoshito Itoh - Grant/Research Support: MSD, Chugai Phamaceutical CO., Bristol-Myers 

Squibb, Dainippon Sumitomo, Otsuka, Takeda, Fujifilm medical, Eisai, 

Mitsubishi Tanabe, AstraZeneca; Speaking and Teaching: MSD 

The following authors have nothing to disclose: Hironori Mitsuyoshi, Kohichiroh 

Yasui, Tasuku Hara, Hiroyoshi Taketani, Hiroshi Ishiba, Akira Okajima, Yuya 

Seko, Atsushi Umemura, Taichiro Nishikawa, Kanji Yamaguchi, Michihisa Moriguchi, 

Yoshio Sumida, Masahito Minami 

887 

Liver MicroRNA-21 is Overexpressed in Non Alcoholic 

Steatohepatitis in Patients and Contributes to this Disease 

In Two Mouse Models by restoring PPARα expression 

Xavier Loyer 1 , Valerie Paradis 3 , Carole Hénique 1 , Anne-Clémence 

Vion 1 , Nathalie Colnot 3 , Coralie L. Guerin 1 , Cécile Devue 1 , Sissi 

On 1 , Jérémy Scetbun 1 , Mélissa Romain 1 , Jean-Louis Paul 4 , Marc 

E. Rothenberg 5 , Patrick Marcellin 2 , Francois Durand 2 , Pierre 

Bedossa 3 , Carina Prip-Buus 6 , Eric Baugé 7 , Bart Staels 7 , Chantal 

Boulanger 1 , Alain Tedgui 1 , Pierre-Emmanuel Rautou 1,2 ; 1 INSERM, 

U970, Paris Cardiovascular Research Center - PARCC@HEGP, 

Paris, France; 2 Hepatology, Hôpital Beaujon, APHP, Clichy, 

France; 3 Pathology, Hôpital Beaujon, APHP, Clichy, France; 4 Service 

de Biochimie, Hôpital Européen Georges Pompidou, AP-HP, 

Paris, France; 5 Cincinnati Children’s Hospital Medical Center, University 

of Cincinnati College of Medicine, Division of Allergy and 

Immunology, Department of Pediatrics, Cincinnati, OH; 6 INSERM, 

U1016, Institut Cochin, Université Paris Descartes, Sorbonne Paris 

Cité, Paris, France; 7 European Genomic Institute for Diabetes 

(EGID); INSERM UMR1011, University of Lille; and Institut Pasteur 

de Lille, Lille, France 

Objective: Previous studies suggested that microRNA-21 

may be up-regulated in the liver in non alcoholic steatohepatitis 

(NASH), but its role in the development of this disease 

remains unknown. This study aimed to determine the role of 

microRNA-21 in NASH. Design: We assessed features of 

NASH in two mouse models of NASH where microRNA-21 

was either pharmacologically inhibited or genetically deleted:


(a) low-density lipoprotein receptor deficient (Ldlr −/− ) mice fed 

a high fat diet and treated with either antagomir-21, antagomir 

control (without specific target) or phosphate buffer saline; 

(b) microRNA-21 deficient and wild-type mice fed a methionine-choline-deficient 

diet (MCD). Wild-type mice fed a chow 

diet served as control. We quantified microRNA-21 levels and 

assessed cell localization in the liver using real-time quantitative 

PCR and in situ hybridization. To determine whether 

the effect of inhibiting or deleting microRNA-21 involved 

PPARα, a known microRNA-21 target implicated in NASH, 

we also fed PPARα deficient mice a MCD diet and treated 

them with either antagomir-21 or antagomir control. Finally, 

microRNA-21 levels were also quantified in the liver of patients 

with NASH or bland steatosis and in normal liver; its cellular 

localization was also determined. Results: Inhibiting or deleting 

liver microRNA-21 expression in both murine models of NASH 

reduced liver cell injury, inflammation, and fibrogenesis without 

affecting liver lipid accumulation or betaoxidation. PPARα 

was decreased in the liver of mice with NASH and restored following 

microRNA-21 inhibition or deletion. In PPARα deficient 

mice fed a MCD diet, antagomir-21 had no effect on liver cell 

injury, inflammation, and fibrogenesis. Liver microRNA-21 was 

overexpressed, primarily in inflammatory and biliary cells, in 

both mouse models as well as in patients with NASH, but not 

in patients with bland steatosis. Conclusion: This study demonstrates 

that microRNA-21 is increased in NASH in liver inflammatory 

cells in mice and in patients. MicroRNA-21 inhibition 

or deletion strongly decreases liver injury, inflammation and 

fibrosis, through PPARα normalization. These findings underscore 

the potential interest of antagomir-21 as a therapeutic 

strategy for NASH. 

Disclosures: 

Jérémy Scetbun - Employment: Merck & Co. 





Abbvie 





The following authors have nothing to disclose: Xavier Loyer, Valerie Paradis, 

Carole Hénique, Anne-Clémence Vion, Nathalie Colnot, Coralie L. Guerin, 

Cécile Devue, Sissi On, Mélissa Romain, Jean-Louis Paul, Marc E. Rothenberg, 

Pierre Bedossa, Carina Prip-Buus, Eric Baugé, Chantal Boulanger, Alain Tedgui 

888 

Two different aspects of PD-1 expression on intrahepatic 

CD4+ and CD8+ T cells in a High Fat High Cholesterol 

Diet-induced murine NASH 

Kanji Yamaguchi 1 , Akira Okajima 1 , Yuya Seko 1 , Hiroshi Ishiba 1 , 

Hiroyoshi Taketani 1 , Atsushi Umemura 1 , Taichiro Nishikawa 1 , 

Yoshio Sumida 1 , Hironori Mitsuyoshi 1 , Kohichiroh Yasui 1 , Takeshi 

Okanoue 2 , Yoshito Itoh 1 ; 1 Gastroenterology, Kyoto Prefectural University 

of Medicine, Kyoto, Japan; 2 Hepatology Center, Saiseikai 

Suita Hospital, Osaka, Japan 

Background: Programmed death (PD)-1 signaling plays an 

important role of the negative regulation of immune responses 

in chronic inflamed liver. Its ligand, PD-L1, is expressed on 

hepatocytes in non-alcoholic steatohepatitis (NASH) livers, and 

that hepatocytes induce apoptosis in T-cells via their interaction. 

The aim of this study is to clarify the pathophysiological 

role of PD-1 expressing CD4+ and CD8+ T cells in murine 

NASH livers. Methods: Eight-week-old male C57/BL6 mice 

were fed either chow or high fat high cholesterol (HFHC) diets 

for 24 weeks. Hepatic lymphocytes were isolated by density 

gradient centrifuge with 35% isotonic Percoll solution after the 

perfusion with 10 ml PBS via portal vein and expression of 

PD-1 was analyzed by FACS at the time points of 8, 16, and 

24 weeks. Moreover, after sorting the fractions of CD4+/PD-1-, 

CD4+/PD-1+, CD8+/PD-1-, CD8+/PD-1+ T cells, mRNA levels 

of TNFα, IL-4, IFNγ, IL-6, foxp3, IL-10 in each group of CD4+ T 

cells and TNFα, IFNγ, granzyme B, perforin, IL-6 in each group 

of CD8+ T cells were assessed by real-time PCR. For further 

study of PD-1 expressing T cells, we analyzed the blocking 

effect of PD-1 signaling on liver injury by continuously using 

PD-1 and PDL-1 neutralizing antibody in HFHC-fed mice for 

24 weeks. Results: The number of PD-1 expressing CD4+ or 

CD8+ T cells was higher in HFHC diet-fed mouse livers and 

increased in a time and the degree of hepatic injury dependent 

manner. Furthermore, HFHC diets significantly induced 

hepatic PD-L1 mRNA at 24 weeks as previously reported. Realtime 

PCR analysis revealed that whereas PD-1 expressions on 

CD4+ T cells suppressed TNFα and IL-6 expression but rather 

increased IL-10 expression, those on CD8+ T cells significantly 

increased granzyme B and perforin despite of decreasing IL-6 

expression. Finally, blockade of PD-1 signaling for 24 weeks 

greatly ameliorated plasma levels of AST and ALT. Conclusion: 

PD-1 expressing T cells were increased in murine NASH livers 

with the elevation of hepatic PD-L1 expression. This regulatory 

molecule may act as a suppressor in CD4+ T cells but an activator 

in CD8+ T cells in this murine NASH model and blockade 

of these PD-1 signaling led to the amelioration of liver injury. 

Disclosures: 






The following authors have nothing to disclose: Kanji Yamaguchi, Akira Okajima, 

Yuya Seko, Hiroshi Ishiba, Hiroyoshi Taketani, Atsushi Umemura, Taichiro Nishikawa, 

Yoshio Sumida, Hironori Mitsuyoshi, Kohichiroh Yasui, Takeshi Okanoue 

889 

DNA Methylation Profiles of Fibrosis-Associated Gene in 

Blood Reflects Liver Fibrosis Severity in Human Nonalcoholic 

Fatty Liver Disease 

Cynthia A. Moylan 1,2 , Susan K. Murphy 3 , Carole Grenier 3 , Manal 

F. Abdelmalek 1 , Anna Mae Diehl 1 ; 1 Duke University Medical Center, 

Durham, NC; 2 Medicine, Durham Veterans Affairs Medical 

Center, Durham, NC; 3 Obstetrics & Gynecology, Duke University, 

Durham, NC 

Background: Nonalcoholic fatty liver disease (NAFLD) can lead 

to cirrhosis, but disease progression is highly variable. Fibrosis 

severity is the only independent predictor of cirrhosis risk in 

NAFLD. DNA methylation and other epigenetic mechanisms 

play a role in instructing fibrosis progression. We reported 

that DNA methylation at specific loci of certain liver genes correlated 

with their expression in liver and with fibrosis severity 

in human NAFLD. Aim: We sought to determine if DNA methylation 

profiles of fibrosis-associated genes can be measured in 

blood and whether they reflect fibrosis severity (i.e., cirrhosis 

risk) in NAFLD patients. Methods: Patients with near-normal histology 

and biopsy-proven NAFLD were selected from the Duke 

NAFLD Biobank. The cohort was stratified into 3 groups as 

shown in the Table: Advanced NAFLD, Mild NAFLD, and Controls. 

DNA was isolated from liver and blood, bisulfite modified 

using the Zymo EZ DNA Methylation Kit, and then analyzed 

by pyrosequencing (PS). Site-specific methylation levels (%met) 

for two genes of interest (FGFR2, cg1385327; CASP1, 

cg13802966) were correlated by tissue type and compared 

by NAFLD severity using Pearson correlation and non-para-


metric t-tests and ANOVA. Results: Age, gender and BMI were 

similar across the 3 groups. More advanced NAFLD patients 

had diabetes mellitus and there were more black patients in the 

control group. A NAFLD Activity Score greater than or equal to 

5 was more common in the advanced NAFLD group. CASP1 

and FGFR2 % met could be measured in both liver and blood. 

CASP1 %met was similar between tissue types and differed 

significantly by group in both liver and blood (controls: 2.8, 

mild NAFLD: 3.1, advanced NAFLD: 3.6; p=0.04). Although 

FGFR2 %met somewhat correlated in paired normal liver and 

blood samples (r=0.55, p=0.01), only FGFR2 %met in liver 

differed significantly by group (controls: 9.1, mild NAFLD: 8.6, 

advanced NAFLD: 4.8; p


quent cellular stress may promote redistribution and retention of 

LC3 in the nuclei. Further studies to explore the mechanism of 

nuclear LC3 redistribution are needed. 

Disclosures: 






The following authors have nothing to disclose: Aaron B. Koenig, Rohini Mehta, 

Gary Bratthauer, Fanny Monge 

Disclosures: 

Brian P. Lam - Advisory Committees or Review Panels: BMS; Speaking and Teaching: 

Gilead; Stock Shareholder: Gilead, Vertex 






The following authors have nothing to disclose: Elzafir Elsheikh, Mehmet Sayiner, 

Zahra Younoszai, Fanny Monge, Lakshmi Alaparthi, Munkhzul Otgonsuren, Hussain 

Allawi, Dinan Abdelatif 

892 

Soluble Intercellular Adhesion Molecule-1 is Independently 

Associated with Increased Collagen Deposition 

in Patients with Nonalcoholic Steatohepatitis 

(NASH) 

Elzafir Elsheikh 1,2 , Mehmet Sayiner 2 , Zahra Younoszai 2 , Fanny 

Monge 1 , Lakshmi Alaparthi 1 , Munkhzul Otgonsuren 2 , Brian P. 

Lam 1 , Hussain Allawi 2 , Dinan Abdelatif 2 , Zachary D. Goodman 1 , 

Zobair M. Younossi 1,2 ; 1 Center for Liver Diseases, Inova Fairfax 

Hospital, Falls Church, VA; 2 Betty and Guy Beatty Center for Integrated 

Research, Inova Health System, Falls Church, VA 

Background: Non-alcoholic fatty liver disease (NAFLD) and its 

progressive subtype NASH, are closely associated with chronic 

inflammation and fibrosis. Circulating soluble adhesion molecules 

are important aspects of this inflammatory response. The 

association of serum circulating soluble adhesion molecules 

in NASH patients with different degree of hepatic collagen 

deposition has not been fully investigated. Aim: To assess the 

relationship between hepatic collagen depositions quantified 

by morphometry and circulating soluble adhesion molecules 

in patients with biopsy-proven NAFLD. Methods: We included 

114 subjects [biopsy-proven NAFLD (n=94) and control subjects 

without NAFLD (n=20)]. Sections of liver biopsies were 

stained with Sirius Red and used for measurement of the percentages 

of collagen with morphometry. Concentrations of the 

adhesion molecules: sICAM-1 (soluble intercellular adhesion 

molecule-1), sVCAM-1 (soluble vascular adhesion molecule-1), 

sP-selectin, and sL-selectin (ng/mL) were determined in the 

serum collected at the time of liver biopsy using Multiplex platform. 

Results: Histologic NASH was seen in 55% of NAFLD. 

There were no significant differences between groups based 

on gender, ethnicity and BMI. sICAM-1 levels were higher in 

NASH group (median, 86) as compared to patients with only 

hepatic steatosis (median, 70, p=0.025 ) or controls without 

NAFLD (median, 65, p=0.033). We also found that, sVCAM-1 

levels were higher in NASH group (median, 409) as compared 

to patients with only hepatic steatosis (median, 357, p=0.025). 

In fact, higher levels of sICAM-1 positively correlated with the 

increased collagen deposition n(r=0.50; p


Giuseppe Penna - Consulting: Intercept Pharmaceuticals; Grant/Research Support: 


The following authors have nothing to disclose: Edina Hot, Ilaria Spadoni, Maria 

Rescigno 

Disclosures: 

The following authors have nothing to disclose: Wilhelmus J. Kwanten, Wim Martinet, 

Benedicte Y. De Winter, Peter P. Michielsen, Sven M. Francque 

894 

Hepatocyte-specific autophagy-deficiency prevents obesity 

in HFD-fed mice and improves glucose tolerance 

Wilhelmus J. Kwanten 1 , Wim Martinet 2 , Benedicte Y. De Winter 

1 , Peter P. Michielsen 1,3 , Sven M. Francque 1,3 ; 1 Laboratory of 

Experimental Medicine and Pediatrics (LEMP) - Gastroenterology 

& Hepatology, University of Antwerp, Wilrijk, Belgium; 2 Laboratory 

of Physiopharmacology, University of Antwerp, Wilrijk, Belgium; 

3 Gastroenterology Hepatology, Antwerp University Hospital 

(UZA), Edegem, Belgium 

Autophagy is a cellular degradation pathway delivering cytoplasmic 

content to lysosomes. There’s increasing evidence that 

autophagy interferes with the pathogenesis of NAFLD. Compared 

to hepatic lipid metabolism, its role in glucose metabolism 

has hardly been studied. AIM To study the influence of 

hepatocellular autophagy on HFD and in glucose metabolism. 

METHODS Hepatocyte-specific autophagy-deficient C57Bl/6J 

mice (Atg7-F) were created using Cre-LoxP with an albumin-promoter 

to delete the autophagy gene Atg7. These mice were 

compared with their controls (Atg7-WT). Mice were fed control 

diet (CD) or high fat diet (HFD) for 16 weeks, with follow-up 

of weight/glycaemia. Glucose (ipGTT) and insulin (ipITT) tolerance 

tests were performed in week 15. RESULTS Atg7-WT/ 

HFD mice show a significant increase in body weight compared 

to Atg7-WT/CD (36.1 vs 27.2g, p


atohepatitis (NASH). We have also shown that nitro-oxidative 

stress might play a key role in the pathogenesis of this dysfunction 

and that activity of the NADPH oxidase is increased in the 

liver of mice with NASH. The aim of this study was to evaluate 

the role played by the NADPHox in the pathogenesis of the 

OXPHOS dysfunction in high fat diet (HFD)-fed mice. Material 

and Methods. Twenty four C57BL/6J mice were distributed in 

four groups: (1) six wild-type (WT) mice fed a standard chow 

diet (SCD) (WT/SCD); (2) six WT mice fed a HFD (WT/HFD); 

(3) six NADPHox deficient mice on a SCD (NOX —/— /SCD); (4) 

six NADPHox deficient mice on a HFD (NOX —/— /HFD). After 

six months on these diets, we studied in the liver: histology, 

MRC activity (spectrophotometry), fully assembled MRC complexes 

(BN-PAGE), subunits of the MRC complexes (BN/SDS- 

PAGE), gene expression of these subunits, as well as of TNFα, 

IFNγ, MCP-1, caspase-3, collagen α1(I), and TGFβ1 (RT-PCR), 

oxidative (thiobarbituric acid-reactive substances, glutathione) 

and nitrosative (immunohistochemistry, nitration of MRC proteins) 

stress, and oxidative DNA damage [8-hydroxy-2ʹ-deoxyguanosine 

(8-OHdG)]. Results: (1) In HFD mice, we found: 

severe steatosis, mild inflammatory infiltrates, ballooning 

degeneration, pericellular fibrosis, 3-tyrosine nitrated proteins, 

and marked increased gene expression of TNFα, IFNγ, 

MCP-1, caspase-3, collagen α1(I), and TGFβ1. Activity of all 

complexes of the MRC was decreased to about 50-60% of 

activity in WT/SCD. Fully assembled complexes were reduced 

to 50% to 60% of that found in control mice. The amount of 

all studied subunits was markedly decreased, particularly, the 

mitochondrial DNA-encoded subunits. Gene expression of 

mitochondrial, but not of genomic, DNA-encoded subunits was 

decreased to about 60% of control gene expression. 8-OHdG 

was increased in mitochondrial, but not in genomic DNA. (2) 

The liver of NOX —/— /HFD mice showed steatosis but not infiltrates, 

ballooning degeneration pericellular fibrosis, 3-tyrosine 

nitrated proteins, or increased gene expression of inflammatory, 

apoptosis or fibrosis related proteins. Likewise, OXPHOS 

activity, subunits, their gene expression, and assembly of these 

subunits in OXPHOS complexes, and 8OHdG were normal in 

the liver of these NOX —/— mice on a HFD. Conclusions. This 

study shows that NADPHox plays a key role in the pathogenesis 

of the OXPHOS dysfunction found in mice on a HFD, as 

this dysfunction was not found in NADPHox deficient mice fed 

a HFD. 

Disclosures: 

The following authors have nothing to disclose: José A. Solís-Herruzo, Pablo 

Solis-Munoz, Daniel Fernández-Moreira, Teresa Muñoz-Yagüe, Inmaculada 

García-Ruiz 

897 

Dual targeting of nuclear receptors ameliorates NAFLD 

pathogenesis in mice 

Pedro M. Rodrigues 2 , Marta B. Afonso 2 , André L. Simão 2 , Marta 

Caridade 2 , Catarina C. Carvalho 3 , Alexandre Trindade 3 , António 

Duarte 3 , Pedro M. Borralho 1 , Mariana V. Machado 4 , Helena Cortez-Pinto 

4,5 , Cecília M. Rodrigues 1 , Rui E. Castro 1 ; 1 iMed.ULisboa 

& Dep. of Biochemistry and Human Biology, Faculty of Pharmacy, 

University of Lisbon, Lisboa, Portugal; 2 iMed.ULisboa, Faculty of 

Pharmacy, University of Lisbon, Lisbon, Portugal; 3 Reproduction 

and Development, Interdisciplinary Centre of Research in Animal 

Health (CIISA), Faculty of Veterinary Medicine, University of Lisbon, 

Lisbon, Portugal; 4 Gastrenterology, Hospital Santa Maria, 

Lisbon, Portugal; 5 Faculty of Medicine, University of Lisbon, Lisbon, 

Portugal 

Non-alcoholic fatty liver disease (NAFLD) burden is rapidly 

increasing and disease pathogenesis remains incomplete. 

We and others have recently characterized miRNAs as novel 

NAFLD pathogenic factors. In addition, nuclear receptors (NRs), 

namely peroxisome proliferator-activated receptors (PPARs) and 

the farnesoid X receptor (FXR) are currently under scrutiny as 

promising therapeutic targets for non-alcoholic steatohepatitis 

(NASH). In fact, the FXR agonist obeticholic acid (OCA) significantly 

ameliorated NASH in a phase 2 clinical trial. We aimed 

to: 1) elucidate the role of miR-21 during NAFLD pathogenesis 

in mice, particularly regarding modulation of the miR-21 target 

PPARα; and 2) evaluate the therapeutic potential of a dual 

NR-targeting approach, specifically activation of PPARα and 

FXR through miR-21 ablation and OCA administration, respectively. 

C57BL/6 wild type (WT; n=24) and miR-21 knockout 

(KO; n=24) mice were fed either a standard diet (SD; n=12) 

or a fast food diet (FF; n=12) for 25 weeks. Six animals from 

each group received OCA (10 mg/kg/day; Intercept Pharmaceuticals, 

Inc.) as a dietary admixture. Human liver biopsies 

were obtained from morbidly obese NAFLD patients at different 

disease stages (n=28). Human and mice liver samples 

were processed for histological analysis and for expression of 

miR-21, pro-inflammatory cytokines, NRs and proteins of the 

insulin signaling pathway, by qRT-PCR and immunoblotting. 

Our results showed that WT FF-fed mice developed hepatomegaly, 

with livers presenting macrovesicular steatosis and 

inflammatory infiltrates, as well as deregulated insulin signaling. 

mRNA levels of TNF-α, IL-6, TLR4, IL-1β and NLRP3 were 

increased, further evidencing development of steatohepatitis. 

miR-21 levels were increased in WT FF-fed mice concomitantly 

with decreased expression of PPARα, a correlation also 

found in NAFLD patients, increasing from steatosis to less and 

more-severe NASH. OCA-fed animals displayed decreased 

CYP7A1 and SREBP-1c liver expression. Further, WT FF+O- 

CA-fed mice exhibited decreased steatosis and miR-21 expression, 

compared with WT FF-fed mice. In addition, KO FF-fed 

mice exhibited significantly reduced liver inflammation, as well 

as steatosis severity, in parallel with increased PPARα, comparing 

with WT FF-fed mice. Significantly, improvement of the 

above histological, biochemical, metabolic and inflammatory 

parameters was augmented in KO FF+OCA-fed mice. In conclusion, 

our results indicate that miR-21 downregulation, leading 

to increased PPARα, together with FXR activation by OCA, 

strongly ameliorate NASH in mice, highlighting the therapeutic 

potential of novel dual-targeting therapies for human NAFLD. 

Disclosures: 



The following authors have nothing to disclose: Pedro M. Rodrigues, Marta B. 

Afonso, André L. Simão, Marta Caridade, Catarina C. Carvalho, Alexandre 

Trindade, António Duarte, Pedro M. Borralho, Mariana V. Machado, Cecília M. 

Rodrigues, Rui E. Castro 

898 

Alteration of Neutrophil-derived Lipocalin 2 in Bone 

Marrow Cells Modulates Non-alcoholic Steatohepatitis 

in Mice 

Dewei Ye 1,2 , Yu Wang 2,1 , Karen S. L. Lam 1,2 , Aimin Xu 1,2 ; 1 State 

Key Laboratory of Pharmaceutical Biotechnology, The University of 

Hong Kong, Hong Kong, Hong Kong; 2 Department of Medicine, 

The University of Hong Kong, Hong Kong, Hong Kong 

Background and Aims: Hepatic infiltration of neutrophils has 

been characterized as one of key histological features of nonalcoholic 

steatohepatitis (NASH) in rodents and humans. Lipocalin-2 

(LCN2) is a secretory glycoprotein originally identified 

from neutrophil granules. This study aims to investigate the 

potential role of LCN2-chimerism in bone marrow-derived cells 

in the pathogenesis of NASH in mice. Methods: LCN2 knockout


(KO) mice were backcrossed into ApoE -/- mice, which are susceptible 

to diet-induced NASH, to generate ApoE-LCN2 double 

knockout (DKO) mice and ApoE -/- /LCN2-wild type (AKO) 

control mice. LCN2-chimeric mice were generated through 

a combined strategy of lethal irradiation and bone marrow 

transplantation. After confirmation of successful reconstitution 

of bone marrow cells, chimeric mice (n= 5 - 6 per group) were 

fed with high fat high cholesterol (HFHC) diet for 12 weeks to 

induce NASH. Results: HFHC diet feeding resulted in a marked 

elevation in hepatic expression and circulating levels of LCN2 

in AKO mice. Transplantation with LCN2-null bone marrow 

cells dramatically abolished HFHC diet-evoked LCN2 elevation 

in both liver and blood in AKO mice, whereas reconstitution of 

wild-type bone marrow cells in DKO mice restored the hepatic 

expression and circulating levels of LCN2. Functionally, transplantation 

of LCN2-expressing bone marrow cells from AKO 

mice into DKO mice was sufficient to restore the susceptibility 

of the recipient mice to HFHC-induced liver inflammation 

and damage, as evidenced by deteriorated liver histology 

and elevated serum activities of alanine aminotransferase and 

aspartate aminotransferase. By contrast, liver lesions triggered 

by HFHC diet were dramatically inhibited in chimeric mice 

bearing LCN2-deficient bone marrow cells. Likewise, HFHC 

diet-elicited hepatic infiltration of inflammatory cells (including 

macrophages and neutrophils) and expression of pro-inflammatory 

cytokines (TNF-α and IL-1β) and chemokines (MCP-1 

and CXCL-2) were significantly augmented in DKO mice by 

transplantation of LCN2-expressing bone marrow cells from 

AKO mice, but were markedly suppressed by reconstitution of 

LCN2-null bone marrow cells from DKO mice into AKO mice 

(P 0.05). Conclusions: Bone marrow-derived cells are the major 

contributor to elevated LCN2 in both circulation and the liver in 

murine model of NASH. LCN2 chimerism in bone marrow-derived 

cells is sufficient to exert significant influence on HFHC 

diet-induced NASH, suggesting that bone marrow-derived 

inflammatory cells and neutrophil-derived LCN2 may serve as 

promising therapeutic target for NASH. 

Disclosures: 

The following authors have nothing to disclose: Dewei Ye, Yu Wang, Karen S. 

L. Lam, Aimin Xu 

899 

Key role of ASMase in the susceptibility of MAT1A deficient 

mice to choline deficient diet-induced steatohepatitis 

Cristina Alarcón-Vila 1 , Vicent Ribas 1 , Susana Nuñez 1 , Carmen 

Garcia-Ruiz 1,2 , Jose Fernandez-Checa 1,2 ; 1 Instituto Investigaciones 

Biomedicas Barcelona, CSIC, Barcelona, Spain; 2 Liver Unit, Hospital 

Clinic-IDIBAPS, Barcelona, Spain 

Non-alcoholic steatohepatitis (NASH) is an advanced stage of 

fatty liver disease characterized by steatosis, oxidative stress, 

fibrosis, inflammation and hepatocellular death, which can 

progress to cirrhosis. Available treatments are ineffective due 

to our poor understanding of the underlined mechanisms. Disturbed 

methionine metabolism (e.g. increased homocysteine 

and decreased S-adenosyl-L-methionine, SAMe) is known to 

contribute to NASH. The first step in methionine metabolism 

is its conversion to SAMe by the enzyme methionine adenosyltransferase, 

MAT1A, which is expressed mainly in adult 

liver. MAT1A deletion markedly depletes hepatic SAMe levels, 

induces NASH and increases liver proliferation. Acid sphingomyelinase 

(ASMase)-induced ceramide generation regulates 

hepatic steatosis and liver fibrogenesis. Our previous observations 

in NASH indicated the association between decreased 

SAMe and ASMase activation. Thus, we investigated the role 

of ASMase in the NASH model of MAT1A null mice fed a 

diet deficient in choline (CD). Methods: MAT1A -/- mice were 

fed a CD diet for 2 weeks with or without amitriptyline (5mg/ 

Kg) to inhibit ASMase or after injection with an adenoviral 

vector expressing ASMase shRNA (ASMSH) to silence hepatic 

ASMase. Moreover, we generated double null mice deficient 

in ASMase and MAT1A to examine their susceptibility to CD 

feeding for 7 days. Serum and liver samples were processed 

to assess ALT levels, ASMase activity, H&E, Oil Red, myeloperoxidase 

staining, inflammation and RT-PCR and Western Blot 

analyses. Results: MAT1A -/- mice fed CD diet and treated with 

amitriptyline or injected with ASMSH exhibited 40-60% lower 

mRNA levels of inflammatory markers such as TNFα, IL-1β and 

IL-6 and less MPO positive cells. Liver damage monitored by 

serum ALT and H&E analyses was reduced in MAT1A KO mice 

after amitriptyline treatment or ASMase silencing (ASMSH). 

Furthermore, markers of endoplasmic reticulum stress PDI, 

CHOP, BiP due to CD diet feeding as well as overexpression of 

lipogenic genes SREBP-1c, FAS, HMGCoA, and SREBP2 and 

fibrosis, Col1A and TGFb, were markedly reduced (50-60%) 

in MAT1A null mice after amitriptyline treatment or ASMase 

silencing. These findings correlated with reduced hepatic steatosis 

monitored by oil-red staining and lower triglycerides 

levels. Similar results regarding steatosis, inflammation, fibrosis 

and liver injury were observed in double knockout mice 

MAT1A -/- /ASMase -/- fed a CD diet. Conclusions: Modulation 

of ASMase activity, through pharmacological o genetic intervention, 

protects MAT1A null mice for developing steatohepatitis, 

highlighting the role of ASMase in the progression of 

steatohepatitis 

Disclosures: 

The following authors have nothing to disclose: Cristina Alarcón-Vila, Vicent 

Ribas, Susana Nuñez, Carmen Garcia-Ruiz, Jose Fernandez-Checa 

900 

Genetic Ablation of Phosphatidylcholine Transfer Protein 

(PC-TP) in ob/ob Mice Improves Glucose Homeostasis 

Without Increasing Energy Expenditure: Evidence for the 

Direct Role for PC-TP in Hepatic Insulin Resistance 

Tibor I. Krisko 1,2 , Katherine B. Leclair 1 , David E. Cohen 1 ; 1 Medicine, 

Brigham and Women’s Hospital, Boston, MA; 2 Medicine, VA 


Background: Hepatic insulin resistance is a hallmark of non-alcoholic 

fatty liver disease (NAFLD). PC-TP is a specific phosphatidylcholine 

binding protein that is highly expressed in liver and 

oxidative tissues. Genetic ablation or small molecule inhibition 

of PC-TP increases both hepatic insulin sensitivity and energy 

expenditure in high fat fed mice. We have demonstrated that 

PC-TP suppresses insulin signaling in the liver. However, its 

direct contribution to hepatic insulin resistance remains unclear 

owing to the capacity of PC-TP to also reduce energy expenditure. 

Aim: We utilized leptin-deficient obese (ob/ob) mice, 

which exhibit both glucose intolerance and defective thermogenesis, 

to dissect the direct contribution of PC-TP to the regulation 

of glucose metabolism. Methods: Mice deficient in both 

leptin and PC-TP were prepared by crossing Pctp -/- mice on a 

C57BL/6J genetic background with ob/ob mice of the same 

background. Tolerance tests to glucose (GTT) were performed 

by measuring blood glucose concentrations at baseline and 

following intraperitoneal administration of glucose. We measured 

energy expenditure by indirect calorimetry as functions 

of variations in ambient temperatures ranging from thermoneutrality 

(30°C) to cold stress (15°C) using a comprehensive 

laboratory animal monitoring system, which also recorded 

physical activity and food intake. Body composition was deter-


mined by NMR spectroscopy. Core body temperature was 

measured by rectal thermocouple probe. Results: As evidenced 

by a decrease in area under the curve (AUC, mg/dl*min), 

ob/ob mice lacking PC-TP demonstrated improvement in the 

GTT (PC-TP-deficient ob/ob 10,846 ± 1229, ob/ob 17,297 ± 

2677, P < 0.05). There were no differences in energy expenditure 

when adjusted for lean body mass at any ambient temperature. 

There were also no effects of PC-TP expression on 

physical activity, food intake or core body temperature. Conclusion: 

Marked improvements in GTT in PC-TP-deficient ob/ob 

mice in the absence of increases in energy expenditure indicate 

a direct role for PC-TP in regulating glucose metabolism. 

These findings support the potential utility of PC-TP inhibitors in 

the management of NAFLD. 

Disclosures: 



The following authors have nothing to disclose: Tibor I. Krisko, Katherine B. 

Leclair 

901 

Aging associated impaired metabolic compensation in 

adipose tissues promotes diet-induced murine nonalcoholic 

fatty liver disease 

Hiroyoshi Taketani 1 , Taichiro Nishikawa 1 , Hisakazu Nakajima 

2 , Satoru Sugimoto 2 , Ikuyo Itoh 2 , Kazuki Koudou 2 , Atsushi 

Umemura 1 , Kanji Yamaguchi 1 , Michihisa Moriguchi 1 , Yoshio 

Sumida 1 , Hironori Mitsuyoshi 1 , Kohichiroh Yasui 1 , Yoshito Itoh 1 ; 

1 Gastroenterology and Hepatology of Kyoto Prefectural University 

of Medicine, Kyoto-shi, Japan; 2 Pediatrics, Kyoto Prefectural University 

of Medicine, Kyoto, Japan 

Background: The prevalence of nonalcoholic fatty liver disease 

(NAFLD), a hepatic manifestation of metabolic syndrome, 

generally increases with age in humans. The progression of 

NAFLD is expected to result from a failed metabolic homeostasis. 

Recently, adipose tissues lead to renewed interest on 

energy metabolism as brown adipose tissues with huge energy 

expenditure was demonstrated to be inducible (iBAT) from 

progenitors within subcutaneous white adipose tissues (sWAT) 

aside from classical BAT (cBAT) in adult human. On the other 

hand, age related change in sWAT is reported to cause a loss 

of iBAT, though its detailed mechanism is still unclear. Here 

we evaluated detailed adipose tissues profiling associated 

with iBAT under aging and its association with progression of 

NAFLD. Methods: Six-week-old male C57BL/6 mice were fed 

with control chow (C) or high-fat diet (60% fat; HF) for 12 or 

24 weeks as Short-term (St) or Long-term (Lt) study group, or 

switched C to HF diet at 18 weeks age (C/HF) for 24 weeks. 

All animals were evaluated on body weight gain, energy 

expenditure and blood biochemical assays including lipid and 

glucose tolerance test. At necropsy liver and adipose tissues 

were examined for histological analysis containing NAFLD 

Activity Score (NAS) and beta-gal staining as a hallmark of 

cellular senescence. The study of gene expression profiling 

on self-renewal of progenitors, iBAT and adipocytokines in 

adipose tissues was also evaluated in Quantitative reverse transcriptase 

polymerase chain reaction and flow cytometry. Inducing 

capacity of BAT from progenitors in adipose tissues was 

also examined in vitro culture system. Results: Animals in St/HF 

group gained body weight, subcutaneous and abdominal adipose 

storages, and showed two types of BAT compensations 

(hypertrophy of classical BAT and appearance of subcutaneous 

induced BAT) and mild hepatic steatosis. On the other 

hand, animals both in Lt/HF and Lt/C/HF groups decreased 

energy expenditure and developed more severe lipid and glucose 

intolerance, higher NAS with lost BAT compensations 

and advanced adipose senescence, where there was no significant 

difference between both groups. Adipose progenitor 

and induced BA associated factors (FGF4, and UCP1) were 

highly expressed in St/HF group with abundant differentiation 

capacity of progenitor cells, and these findings disappeared in 

aged groups (Lt/HF and Lt/C/HF). Conclusion: Aging caused 

impaired metabolic homeostasis and progression of NAFLD 

through deteriorated adipose compensation. Aging associated 

adipose tissues profiling might provide a new insight to go 

toward a fully-integrated understanding of NAFLD 

Disclosures: 






The following authors have nothing to disclose: Hiroyoshi Taketani, Taichiro 

Nishikawa, Hisakazu Nakajima, Satoru Sugimoto, Ikuyo Itoh, Kazuki Koudou, 

Atsushi Umemura, Kanji Yamaguchi, Michihisa Moriguchi, Yoshio Sumida, 

Hironori Mitsuyoshi, Kohichiroh Yasui 

902 

The dual FXR/TGR5 agonist INT-767reduces visceral fat 

mass, promoting preadipocyte brown differentiation, 

mitochondrial function and insulin sensitivity in a rabbit 

model of high fat diet-induced metabolic syndrome 

Linda Vignozzi 2 , Ilaria Cellai 2 , Sandra Filippi 1 , Paolo Comeglio 2 , 

Tommaso Mello 2 , Daniele Bani 3 , Daniele Guasti 3 , Erica Sarchielli 3 , 

Annamaria Morelli 3 , Elena Maneschi 2 , Gabriella Barbara Vannelli 

3 , Luciano Adorini 4 , Mario Maggi 2 ; 1 Interdepartmental Laboratory 

of Functional and Cellular Pharmacology of Reproduction, 

Dep. of NEUROFARBA and Dep. of Experimental and Clinical 

Biomedical Sciences, University of Florence, Florence, Italy; 2 Dep. 

of Experimental and Clinical Biomedical Sciences, University of 

Florence, Florence, Italy; 3 Dep. of Experimental and Clinical Medicine, 

University of Florence, Florence, Italy; 4 Intercept Pharmaceuticals, 

New York, NY 

Expanding brown adipose tissue is a potential therapeutic strategy 

to counteract insulin resistance and metabolic syndrome 

(MetS).Farnesoid X receptor (FXR) and Takeda G protein-coupled 

receptor 5 (TGR5)activation enhances insulin sensitivity, 

suggesting the capacity of FXR/TGR5 agonists to promote 

brown differentiation in adipose tissue. Treatment with increasing 

doses of the dual FXR/TGR5 agonist INT-767 (3,10,30mg/ 

Kg, orally, daily, 5 days a week for 12 weeks) in a rabbit model 

of high fat diet (HFD)-induced MetS, characterized by insulin 

resistance, hypertension, atherogenic dyslipidemia and visceral 

adipose tissue (VAT) accumulation, dose-dependently reduced 

VAT mass, glycemia, insulin resistance and cholesterol levels 

while significantly increasing HDL levels. INT-767 (30mg/Kg/ 

day) also significantly reduced HFD-induced hepatomegaly and 

ALT increase. Treatment with INT-767 significantly decreased 

HFD-induced adipocyte hypertrophy and reduced GLUT4 translocation 

to the plasma membrane, both considered hallmarks 

of insulin resistance in VAT. Treatment with INT-767 also significantly 

induced VAT expression of genes related to nitric oxide 

(NO)/cyclic guanosine-3’,5’-monophosphate (cGMP)/protein 

kinase G (PKG) signaling, mediating both mitochondriogenesis 

and brown adipocyte differentiation. Rabbit preadipocytes 

(rPADs), isolated from the different experimental groups, were 

investigated for their spontaneous adipogenic potential. The 

expression of genes specific for: (i) brown fat (UCP1, CIDEA, 

BMP4, BMP7, TMEM26,HOXC9, and LHX8),(ii) mitochondrial 

biogenesis (Tfam, NRF1,GCa1, GCb1, PKG1), (iii)membrane 

respiratory chain (SLC25A12, NDUFB3, NDUFB5, SDHB,


(iv) pro-fusion (Mfn2) and pro-fission (Fis1) proteins of mitochondria, 

were all significantly increased in rPADs from INT- 

767-treated compared to HFD rabbits. Transmission electron 

microscopy demonstrated that INT-767 treatment normalized 

HFD-induced reduction of mitochondrial cristae. INT-767 treatment 

was also able to: (i) improve mitochondrial architecture 

and dynamic, with the majority of mitochondria continuously 

moving and changing shape, as assessed by time lapse imaging 

with mitochondria-targeted fluorescent probe MitoTracker, 

(ii) reduce superoxide production, assessed by measuring the 

time-dependent accumulation of dihydroethidium-derived fluorescence, 

(iii) improve insulin sensitivity assessed by measuring 

3 H-2-deoxy-D-glucose uptake in response to increasing insulin 

concentrations. In conclusion, the dual FXR/TGR5 agonist INT- 

767 ameliorates the metabolic profile and reduces visceral 

adiposity by improving insulin sensitivity and promoting brown 

differentiation in visceral adipose tissue. 

Disclosures: 


Mario Maggi - Consulting: BAYER, ELI LILLY, MENARINI, PROSTRAKAN, INTER- 

CEPT 

The following authors have nothing to disclose: Linda Vignozzi, Ilaria Cellai, 

Sandra Filippi, Paolo Comeglio, Tommaso Mello, Daniele Bani, Daniele Guasti, 

Erica Sarchielli, Annamaria Morelli, Elena Maneschi, Gabriella Barbara Vannelli 

903 

CXCL1 promotes steatohepatitis and fibrosis in a mouse 

model of high-fat diet-plus-binge ethanol feeding 

Zhou Zhou 1 , Ming-Jiang Xu 1 , Binxia Chang 1,2 , Yan Cai 1 , Bin 

Gao 1 ; 1 Laboratory of Liver Diseases, National Institute of Alcohol 

Abuse and Alcoholism, Rockville, MD; 2 Diagnosis and Treatment 

Center for Non-Infectious Liver Diseases, Institute of Alcoholic Liver 

Disease, Beijing 302 Hospital, Beijing, China 

Background and Aims: Obesity and alcohol abuse are independent 

risk factors of human liver diseases. Although some 

epidemic studies suggested a synergistic effect, the exact function 

and mechanism of liver lipid accumulation and alcohol 

intake on liver injury is elusive. Methods: In our present study 

we developed a murine model of high fat diet (HFD)-plus-binge 

ethanol feeding. By using this model, we examined the role of 

CXCL1 in steatohepatitis and fibrosis. Results: This HFD-plusbinge 

ethanol feeding model induced significant steatohepatitis 

and liver fibrosis, as demonstrated by increased expression 

of collagen genes and obvious collagen deposition shown in 

Sirius Red staining. When we analyzed the mechanisms, we 

found that HFD-plus-binge resulted in a significant increase 

of neutrophil infiltration in the liver by flow cytometry analysis 

of Gr-1 hi CD11b + cells and immunohistochemistry staining of 

MPO. Hepatic and serum CXCL1 levels were highly elevated 

after HFD-plus-binge ethanol feeding. This upregulation was 

mediated by free fatty acid accumulation and the downstream 

ERK, JNK and NF-kB pathways. Blocking CXCL1 by genetic 

disruption or a neutralizing antibody improved steatohepatitis. 

In contrast, overexpressing CXCL1 in hepatocytes with adenovirus 

induced significant liver inflammation and injury. Moreover, 

hepatocyte derived CXCL1 also contributed to liver fibrosis. 

Overexpression of CXCL1 in hepatocytes devastated collagen 

deposition in HFD mice, while neutralizing CXCL1 ameliorated 

fibrosis induced by HFD-plus-binge ethanol feeding. Conclusions: 

Our results revealed a pivotal role of hepatocyte derived 

CXCL1 in the liver damage and fibrosis caused by fatty liver 

disease and alcohol abuse. Inhibiting the production of CXCL1 

or blocking its function might be a useful therapeutic strategy in 

preventing alcoholic hepatitis and the related fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Zhou Zhou, Ming-Jiang Xu, Binxia 

Chang, Yan Cai, Bin Gao 

904 

Vitamin D through Induction of Paneth Cell Defensins 

Attenuates Gut Dysbiosis and Improves Metabolic Disorders 

in Animal Models 

Yuan-Ping Han 1 , Danmei Su 1 , Yuanyang Nie 1 , Airu Zhu 1 , Zishuo 

Chen 1 , Li Zhang 1 , Pengfei Wu 1 , Mei Luo 1 , Guihui Wu 2 , Richard 

Hu 3 , Aurelia Lugea 4 , Jun Xu 5 , Hidekazu Tsukamoto 5 , Stephen J. 

Pandol 4 ; 1 College of Life Sciences, Sichuan University, Chengdu 

City, China; 2 Chengdu Public Health Clinical Center, Chengdu, 

China; 3 Olive View-UCLA Medical Center, Los Angeles, CA; 

4 Cedars-Sinai Medical Center, Los Angeles, CA; 5 University of 

Southern California, Los Angeles, CA 

Metabolic syndrome (MetS) and vitamin D insufficiency/deficiency 

are co-prevalent in general population worldwide, but 

their causal relationship and mechanisms remain to be determined. 

Large body of clinical surveys shows vitamin D insufficiency/deficiency 

is associated with type-2 diabetes, obesity, 

non-alcoholic steatohepatitis (NASH), and metabolic disorders. 

Our previous work also demonstrated dietary vitamin D deficiency 

promotes the high fat feeding initiated NASH, in part 

through impaired enterohepatic circulation. Since vitamin D 

receptor (VDR) is highly expressed in ileum epithelia, we tested 

the role of vitamin D signaling in maintaining intestinal integrity 

and eubiosis. Balb/C and C57/B6 mice were fed for 4-5 

months by (1) AIN93 chow with vitamin D 3 

supplementation 

as control, or (2) AIN93 with vitamin D depletion, VDD, or (3) 

high fat (60% calorie) with vitamin D supplementation, HFD, 

and (4) high fat chow with vitamin D depletion, HFD+VDD. 

Our results showed that HFD alone was not sufficient to induce 

NASH and metabolic disorders, but additional dietary vitamin 

D deficiency (HFD+VDD) resulted in severe NASH and insulin 

resistance. The interface of ileum—being defined by the intestinal 

epithelia and the adjacent gut microbiota in the lumen— 

was impaired by the mice fed with HFD+VDD. Specifically, the 

Paneth cell-specific defensins including a-defensin 5 (DEFA5), 

MMP7 which activates defensins, tight junction genes, and 

goblet cell produced mucous protein MUC2 were all thoroughly 

suppressed or down regulated in the ileum, resulting 

in mucosal collapse, gut leakiness, dysbiosis, endotoxemia, 

systemic inflammation, insulin resistance, hepatic steatosis, 

and metabolic disorders. Moreover, Helicobacter hepaticus, 

a known murine hepatic-pathogen causing chronic hepatitis, 

was substantially amplified in within the ileum of the mice fed 

with HFD+VDD, while the beneficial symbiotic Akkermansia 

muciniphila was diminished. Mice deficient in VDR exhibited 

spontaneous NASH and very similar dysbiosis to the mice fed 

with HFD+VDD. Remarkably, oral administration of DEFA5 to 

the metabolic mice restored gut eubiosis, showing suppression 

of Helicobacter hepaticus and reviving Akkermansia muciniphila 

within the lumen of ileum, where VD signaling and DEFA5 

were impaired by VDD. Along with rebalancing of gut microbiota, 

oral administration of DEFA5 resolved hepatic steatosis 

in the HFD+VDD fed mice. These results reveal the critical roles 

of vitamin D/VDR pathway in optimal expression of defensins, 

which in turn regulate intestinal integrity and eubiosis to protect 

the host from metabolic disorders including NASH. 

Disclosures: 

Stephen J. Pandol - Consulting: Calcimedica, Shire, Takeda; Grant/Research 

Support: NIH, Department of Veterans Affairs, Calcimedica; Stock Shareholder: 

GIRx, Phyteau, Pandol Bros. Inc., Ranch 50


The following authors have nothing to disclose: Yuan-Ping Han, Danmei Su, 

Yuanyang Nie, Airu Zhu, Zishuo Chen, Li Zhang, Pengfei Wu, Mei Luo, Guihui 

Wu, Richard Hu, Aurelia Lugea, Jun Xu, Hidekazu Tsukamoto 

905 

Fecal Alkaline Phosphatase Is a Determinant Of Metabolic 

Endotoxemia And Severity Of Nonalcoholic Fatty 

Liver Disease (NAFLD) 

Kalyani Daita 1 , Lorenzo Mulazzani 2 , Robert Vincent 1 , Amon 

Asgharpour 1 , Sophie C. Cazanave 1 , Mohammad S. Siddiqui 1 , 

Puneet Puri 1 , Michael Idowu 1 , Sherry L. Boyett 1 , Faridoddin Mirshahi 

1 , Hae-Ki Min 1 , Robert E. Brown 3 , Masoumeh Sikaroodi 3 , 

Patrick M. Gillevet 3 , Arun J. Sanyal 1 ; 1 VCU, Richmond, VA; 2 Università 

di Bologna, Bologna, Italy; 3 George mason university, 

Manassas, VA 

BACKGROUND: Endotoxemia plays a key role in the genesis of 

insulin resistance (IR) and activation of the innate immune system 

(IAS) in the metabolic syndrome (Nature 2012; 485:S12). 

IR and activation of the IAS are important in the pathogenesis 

of NAFLD. Alkaline phosphatase (AP) dephosphorylates 

endotoxin (ET) and renders it inactive and intestinal AP (IAP) 

is critical to limit ET exposure (AmJPath151.4;1997,1163). 

HYPOTHESIS: We hypothesized that fecal IAP (F-IAP) activity 

would be inversely related to severity of endotoxemia and the 

severity of underlying NAFLD. AIMS: (1) to determine if NASH 

was associated with lower fecal IAP and higher endotoxin levels 

compared to controls and NAFL, (2) to determine the relationship 

between fecal IAP and plasma ET and liver histology 

across the spectrum of NAFLD, (3) to determine if changes in 

fecal IAP were linked to intestinal inflammation as assessed by 

fecal calprotectin, and (4) to identify changes in the intestinal 

microbiome associated with altered fecal IAP. METHODS: Subjects 

with nonalcoholic fatty liver (NAFL, n= 9) or nonalcoholic 

steatohepatitis (NASH, n=15) were compared to 5 weight 

matched controls. Fecal IAP was measured by chromogenic 

quantification with Dako 5-bromo-4-chloro-3-indoyl-phosphate/ 

nitroblue with calf AP as an internal control. Plasma ET and 

fecal calprotectin were measured by a chromogenic quantification 

kit and ELISA respectively. Liver histology was scored 

according to NASH CRN criteria. Stool microbiome was analyzed 

using 16s pyrosequencing. RESULTS: F-IAP was significantly 

lower in NASH compared to NAFL as well as controls 

(237.13 U/mg dry weight vs 306.03 and 314.2, p< 0.01 

and 0.03 respectively). The plasma AP was similar between 

groups. NASH was associated with higher endotoxemia than 

both NAFL and controls (1.53 EU/ml vs 1.3 and 1.26, p< 

0.02 and 0.03 respectively). There was a tight inverse relationship 

between F-IAP and ET levels (r=-0.42, p


Disclosures: 





The following authors have nothing to disclose: Yusuke Ebisutani, Yoshihiro 

Kamada, Sachiho KIda, Kayo Mizutani, Maaya Akita, Akiko Yamamoto, 

Hironobu Fujii, Tomoaki Sobajima, Naoko Terao, Shinji Takamatsu, Yuichi 

Yoshida, Eiji Miyoshi 

907 

High Milkfat Diet Causes More Severe NASH Compared 

to High Lard Diet: The Dual FXR-TGR5 Agonist INT-767 

Prevents NASH in both Models 

Xiaoxin Wang 1 , Yuhuan Luo 1 , David J. Orlicky 2 , Luciano Adorini 3 , 

Moshe Levi 1 ; 1 Medicine, Univ Colorado, Aurora, CO; 2 Pathology, 

University of Colorado, Aurora, CO; 3 Intercept Pharmaceuticals, 

New York, NY 

Diets enriched with saturated fatty acids may cause increased 

incidence of obesity, insulin resistance, nonalcoholic fatty liver 

disease (NAFLD) and nonalcoholic steatohepatitis (NASH). In 

this study in C57BL/6J mice we compared the effects of high 

milkfat diet (60 kcal% milkfat) versus high lard fat diet (60 

kcal% lard fat). We found that both diets induced equal degree 

of weight gain, increased fasting glucose, increased serum 

insulin, and increased serum cholesterol levels. However, 

following a milkfat diet in the liver more inflammatory foci, 

lipogranulomas, ballooned cells, and fibrosis were observed. 

Oral treatment with the dual FXR-TGR5 agonist INT-767, 10 

mg/kg bw/day, or 30 mg/kg bw/day, or 100 mg/kg bw/ 

day, starting from 8 weeks of age and continuing for 8 months, 

in a dose dependent way significantly decreased microsteatosis, 

inflammation, fibrosis, and liver cell injury. In mice fed 

the milkfat and lard diets, treatment with 100 mg/kg bw/day 

of INT-767 was able to abrogate, in all mice tested, hepatic 

macrosteatosis, microsteatosis, as well as fibrosis, decreasing 

them to control levels (0%). However, in contrast to mice fed the 

high lard fat diet where treatment with 100 mg/kg bw/day of 

INT-767 was able to decrease inflammation to 0%, in mice fed 

the milkfat diet 33% of the livers still demonstrated evidence of 

inflammation. INT-767 is therefore able to effectively prevent 

NASH in two different models of diet induced obesity and 

insulin resistance. 

Disclosures: 




The following authors have nothing to disclose: Xiaoxin Wang, Yuhuan Luo, 


908 

Ornithine transcarbamylase gene expression and 

hepatic urea nitrogen handling are reduced in models 

of NAFLD and recovers with dietary modulation and 

reducing bacterial translocation: Rationale for ammonia 

lowering therapy in NASH patients 

Karen Louise Thomsen 1 , Francesco De Chiara 1 , Fausto Andreola 1 , 

Krista Rombouts 1 , Jane MacNaughtan 1 , Jane Grove 2 , Guruprasad 

P. Aithal 2 , Rajeshwar Mookerjee 1 , Rajiv Jalan 1 ; 1 Institute of Liver 

and Digestive Health, University College London, London, United 

Kingdom; 2 Nottingham Digestive Diseases Biomedical Research 

Unit, Nottingham University Hospital, Nottingham, United Kingdom 

Background: Non-alcoholic fatty liver disease (NAFLD) is a 

spectrum of liver disease ranging from steatosis, through non-alcoholic 

steatohepatitis (NASH) to cirrhosis. In NASH, bacterial 

translocation and dysfunctional mitochondria may affect the 

function of the mitochondrial urea cycle enzyme ornithine transcarbamylase 

(OTC), and result in hyperammonemia, which 

has been shown to activate hepatic stellate cells in vivo and in 

vitro. This may favour the progression of NAFLD. The aims of 

this study were to determine whether gene and protein expression 

and function of OTC are altered in animal models of NASH 

and NAFLD patients. We also determined if this was reversible 

with recovery of animals by restoring the diet and by reducing 

bacterial translocation. Methods: Two animal models of NASH 

were studied. (a) high-fat, high-cholesterol diet (HFC) for 10 

months and then recovery for 2 months (b) methionine-choline 

deficient diet (MCD) for 4 weeks treated with or without Yaq- 

001 (Yaqrit Ltd.), a nanoporous carbon which has been shown 

to reduce bacterial translocation. In humans, we obtained liver 

biopsies from 16 NAFLD patients during bariatric surgery and 

measured the OTC gene expression. Results: In both animal 

models, gene and protein expression of OTC was reduced 

significantly. In the HFC rats, reversal of NASH by changing 

the diet to normal chow restored OTC gene (0.53 (0.41-0.68) 

vs. 0.32 (0.28-0.37), P


fourth week and twice at 1-week intervals to investigate their 

importance in the pathogenesis. Functional effects of Tregs on 

lipid metabolism, oxidative stress, and macrophage activation 

were then examined. Results: Chronic alcohol feeding induced 

liver steatosis and increased serum alanine aminotransferase. A 

decrease of hepatice Tregs was observed in these mice. Adaptive 

transfer of Tregs was able to abolish hepatic inflammation 

and liver injury. Adaptive transfer of Tregs downregulated the 

enhanced expression of nuclear Sterol regulatory element-binding 

protein 1c (SREBP1c) and its downstream genes induced 

in alcohol-fed mice. This intervention also ameliorated hepatic 

oxidative stress, increased the level of phosphorylatied adenosine 

monophosphate-activated protein kinase (AMPK), peroxisome 

proliferator-activated receptor (PPAR) α in the nucleus, 

and its downstream genes involved in fatty acid metabolism, 

which were inhibited by alcohol-feeding. Furthermore, Tregs 

inhibited MCP-1 and TNF- overproduction and macrophage 

activation in the liver. In vitro, Tregs suppressed the expression 

of MCP-1, TNFα, and CD14 on monocytes/macrophages that 

underwent LPS and alcohol co-treatment. This suppression was 

markedly abrogated by neutralizing anti-IL-10 mAbs. Conclusions: 

Our results indicate that Tregs suppress the development 

of AFL, in part through modulating lipid metabolism, oxidative 

stress, and the macrophage pro-inflammatory response. 

Disclosures: 

Qin Ning - Advisory Committees or Review Panels: ROCHE, NOVARTIS, BMS, 

MSD, GSK; Consulting: ROCHE, NOVARTIS, BMS, MSD, GSK; Grant/Research 

Support: ROCHE, NOVARTIS, BMS; Speaking and Teaching: ROCHE, NOVAR- 

TIS, BMS, MSD, GSK 

The following authors have nothing to disclose: Hongwu Wang, Ting Wu, Yaqi 

Wang, Xiaojing Wang, Xiaoping Luo 

910 

NOX2 deficiency attenuates ethanol-induced hepatic 

steatosis by down-regulating CB1R-mediated lipogenesis 

in mice 

Jong-Min Jeong, So Yeon Kim, Won-IL Jeong; KAIST, Daejeon, 


Background: Lines of evidence have suggested that alcohol-mediated 

oxidative stress plays important roles in hepatic 

steatosis, in which hepatic stellate cells (HSCs) stimulate CB1 

receptor (CB1R)-mediated hepatic lipogenesis by producing 

an endocannabinoid, 2-arachidonoylglycerol (2-AG). However, 

it is not clear yet whether oxidative stress has an effect 

on endocannabinoid/CB1R signaling pathway. In the present 

study, we investigated the role of NADPH oxidase 2 (NOX2) in 

hepatic CB1R signaling pathway during alcoholic steatosis in 

mice. Methods: WT and NOX2 deficient (NOX2 -/- ) mice were 

fed with liquid ethanol diet for 8 weeks. Sera and liver tissues 

of mice were used for biochemical analyses, hitopathological 

observation, fluorescence-activated sorting (FACS) analysis 

and real-time PCR analyses. LPS treatment, NOX2 siRNA, measurement 

of reactive oxygen species (ROS), immunoblotting 

and immunostaining were performed in primary hepatic macrophages 

and Raw 264.7 cell lines. Results: In histological findings, 

livers of NOX2 -/- mice showed less hepatic steatosis than 

WT mice. In FACS analyses, populations of Gr1 + CD11b + and 

F4/80 + CD11b + cells in WT mice were similar with those of 

NOX2 -/- mice. However, gene expression of inflammatory cytokines 

in WT mice was significantly higher than that of NOX2 - 

/- 

mice. In addition, LPS-mediated ROS generation in primary 

hepatic and Raw 264.7 macrophages was remarkably attenuated 

as NOX2 and toll-like receptor 4 (TLR4) were depleted in 

macrophages, suggesting that TLR4 activation was implicated 

in NOX2-mediated ROS generation. Furthermore, in co-culturing 

macrophages with HSCs, we found that NOX2-mediated 

ROS generation in hepatic macrophages increased gene 

expression of diacylglycerol lipases, specific enzymes for 2-AG 

production, in HSCs, leading to enhance hepatic lipogenesis 

through CB1R signaling in hepatocyte. Conclusions: Based on 

our data, LPS-mediated NOX2 activation in hepatic macrophage 

played a decisive role on alcoholic hepatic steatosis by 

stimulating endocannabinoid production of HSCs and CB1R 

signaling in hepatocyte. Therefore, NOX2 might be a therapeutic 

candidate for alcoholic liver disease. 

Disclosures: 

The following authors have nothing to disclose: Jong-Min Jeong, So Yeon Kim, 

Won-IL Jeong 

911 

L-Fabp deletion attenuates both hepatic steatosis and 

fibrosis resulting from hepatic microsomal triglyceride 

transfer protein (Mttp) deletion 

Elizabeth P. Newberry 2 , Susan M. Kennedy 2 , Yan Xie 2 , Hui 

Jiang 2 , Anping Chen 1 , Daniel S. Ory 2 , Nicholas O. Davidson 2 ; 

1 St. Louis University, St Louis, MO; 2 Washington University School 

of Medicine, St Louis, MO 

Genetic or pharmacological inhibition of hepatic microsomal 

triglyceride transfer protein (Mttp) blocks VLDL secretion 

causing hepatic steatosis. However, the effects on fibrosis are 

unknown. Here we asked if liver specific (Alb Cre) Mttp deletion 

(Mttp-LKO) promotes hepatic stellate cell activation and 

fibrosis and if this phenotype is altered by germline L-Fabp 

deletion (DKO mice). Mttp-LKO mice fed low fat chow showed 

~10 fold increased hepatic triglyceride (TG) vs C57BL/6J controls 

at 12 weeks, with upregulated expression of fibrotic genes 

(Col1a1, αSMA, Ctgf) and increased collagen staining (C57, 

4.1 ± 0.6 foci/field; Mttp-LKO, 10.1 ± 0.9, n=5-7/group). 

Steatosis was significantly attenuated in DKO mice (223 ± 13 

mg TG/mg protein vs 331 ± 36, Mttp-LKO, n=6), with reduced 

expression of fibrogenic mRNAs and decreased fibrotic staining 

(DKO, 5.1 ± 0.5 foci/field). High fat diet fed Mttp-LKO 

mice also exhibited 2.5-fold increased hepatic TG, which 

was attenuated in DKO mice, again with attenuated Col1a1 

gene expression. Expression of lipid droplet proteins Pln2 and 

Cidec mRNA were reduced in DKO vs Mttp-LKO, consistent 

with reduced steatosis. In vitro FA trafficking revealed ~25% 

decreased incorporation of FA into TG in DKO hepatocytes, 

yet reduced FA oxidation compared to Mttp-LKO, suggesting 

altered FA compartmentalization. ER stress response (Xbp1s, 

Atf4, Ern1) was decreased ~40% in DKO livers vs both C57 

and Mttp-LKO. Oxidative stress, assessed by lipoperoxide levels, 

was increased 4-fold in Mttp-LKO vs DKO (Mttp-LKO, 6.8 

± 0.9 nmol LPO/mg protein; DKO, 1.7 ± 0.3, n=4). To understand 

the basis for reduced fibrosis in mice lacking L-Fabp, we 

undertook broad range lipidomic analyses in livers of Mttp- 

LKO and DKO mice. As expected, TG mass was decreased 

~25% in DKO, with a disproportionate reduction in TG species 

containing essential fatty acids C18:2 and C18:3 (Mttp-LKO, 

11.9 ± 0.6; DKO, 8.0 ± 0.6, n=4-5/group, p


a risk for hepatic fibrosis, a concern in light of Phase III clinical 

trials of pharmacologic Mttp inhibitors. 

Disclosures: 

The following authors have nothing to disclose: Elizabeth P. Newberry, Susan 

M. Kennedy, Yan Xie, Hui Jiang, Anping Chen, Daniel S. Ory, Nicholas O. 

Davidson 

912 

intrahepatic expression levels of bile acid transporters 

are inversely correlated with histological progression of 

non-alcoholic fatty liver disease 

Kazuya Okushin 1 , Takeya Tsutsumi 2 , Kenichiro Enooku 1 , Hidetaka 

Fujinaga 1 , Akira Kado 1 , Kyoji Moriya 3 , Hiroshi Yotsuyanagi 2 , 

Kazuhiko Koike 1 ; 1 Department of Gastroenterology, Graduate 

School of Medicine, The University of Tokyo, Tokyo, Japan; 

2 Department of Infectious Diseases, Graduate School of Medicine, 

The University of Tokyo, Tokyo, Japan; 3 Department of Infection 

Control and Prevention, Graduate School of Medicine, The University 

of Tokyo, Tokyo, Japan 

Background & Aims Non-alcoholic fatty liver disease (NAFLD) 

is increasing in the world including Asian countries. NAFLD 

contains a disease spectrum ranging from simple steatosis to 

nonalcoholic steatohepatitis (NASH). The latter is considered 

as a progressive disease, of which pathogenesis remains 

largely unclear. Recently, bile acid (BA) metabolism is focused 

as a therapeutic target of NASH. The aim of this study was to 

identify changes of bile acid metabolism in NAFLD patients 

in terms of disease progression. Methods From November 

2011 to June 2014, we prospectively enrolled patients with 

clinically suspected NAFLD at The University of Tokyo Hospital. 

Patients taking ursodeoxycholic acid were excluded in this 

study. Disease progression was evaluated by NAFLD activity 

score (NAS). Intrahepatic expression levels of genes related 

to BA metabolism were determined by quantitative PCR and 

immunohistochemistry. This study was approved by the ethics 

committees of The University of Tokyo and all patients gave 

informed written consent for the study according to the Declaration 

of Helsinki. Results Seventy-eight patients (male: female 

= 49: 29) histologically diagnosed as NAFLD by significant 

steatosis, namely the percentage of hepatocytes showing fatty 

changes was ≥ 5% and coincidence with Matteoni classification, 

were analyzed. Expression levels of farnesoid X receptor 

(FXR) and liver receptor homolog 1 (LRH1), key molecules 

associated with BA metabolism, were significantly decreased 

along with NAS increase only in female. Small heterodimer 

partner (SHP), a downstream transcriptional repressor of FXR, 

was similarly changed but only in male. On the other hand, 

the level of cholesterol 7 alpha-hydroxylase (CYP7A1), a key 

enzyme of BA synthesis, was not changed with the elevation of 

NAS in both genders. However, expression levels of an export 

transporter, bile salt export pump (BSEP), and an uptake transporter, 

Na + /taurocholate cotransporter (NTCP), were significantly 

down-regulated as the NAS increases in both genders. 

Another export transporter, multidrug resistance-associated 

protein 2 (MRP2) was also significantly down-regulated but 

only in female. Decreases in protein levels were confirmed by 

immunohistochemistry on both BSEP and MRP2. Conclusions 

Expression levels of BA export transporters, BSEP and MRP2, 

were inversely correlated with NAS in the liver of NAFLD 

patients. This down-regulation are supposed to cause excessive 

increase of BA levels in hepatocyte, leading to hepatocyte injuries. 

Although the mechanism underlying the down-regulation 

remains to be elucidated, our findings in patients might lead to 

the development of new therapeutic options for NASH. 

Disclosures: 

The following authors have nothing to disclose: Kazuya Okushin, Takeya Tsutsumi, 

Kenichiro Enooku, Hidetaka Fujinaga, Akira Kado, Kyoji Moriya, Hiroshi 

Yotsuyanagi, Kazuhiko Koike 

913 

Non-invasive quantitative decline in liver fat content on 

MRI and histologic response in nonalcoholic steatohepatitis: 

A secondary analysis of MOZART trial 

Janki R. Patel 1 , Ricki Bettencourt 2,3 , Jeffrey Y. Cui 2 , Joanie 

Salotti 4,2 , Jonathan Hooker 5 , Archana Bhatt 2 , Carolyn Hernandez 

2 , Phirum Nguyen 2 , Hamed Aryafar 5 , William Haufe 5 , Catherine 

A. Hooker 5 , Lisa M. Richards 2,4 , Claude B. Sirlin 5 , Rohit 

Loomba 2,4 ; 1 Department of Internal Medicine, University of California, 

San Diego, La Jolla, CA; 2 NAFLD Translational Research 

Unit, University of California, San Diego, La Jolla, CA; 3 Division of 

Epidemiology, University of California, San Diego, La Jolla, CA; 

4 Division of Gastroenterology, Department of Medicine, University 

of California, San Diego, La Jolla, CA; 5 Liver Imaging Group, 

Department of Radiology, University of California, San Diego, La 

Jolla, CA 

Background: Magnetic resonance imaging-estimated proton 

density fat fraction (MRI-PDFF) has been shown to be a non-invasive, 

accurate and reproducible imaging-based biomarker 

for assessing steatosis change and treatment response in nonalcoholic 

steatohepatitis (NASH) clinical trials. However, there 

are no data on the amount of decline in liver fat shown on MRI- 

PDFF that corresponds to histologic response in the setting of a 

clinical trial in NASH. We aimed to quantitatively compare the 

amount of change in liver fat using MRI-PDFF between histologic 

responders vs. histologic non-responders. Methods: This study is 

a secondary analysis of the MOZART trial, a randomized, placebo-controlled, 

double-blind trial, which included 50 patients 

with biopsy-proven NASH to assess efficacy of ezetimibe 10 

mg orally daily in reducing liver fat as measured by MRI-PDFF. 

All patients enrolled in the trial underwent biochemical testing, 

liver biopsy, and MRI at baseline and of those who had both 

liver biopsy and MRI-PDFF at weeks 0 and 24 were included in 

this analysis. Patients were classified as histologic responders 

if they had ≥2 point reduction in NAFLD Activity Score (NAS) 

without any increase in fibrosis stage and as histologic non-responders 

if they did not meet this criterion. We performed a 

head-to-head comparative analysis of histologic responders 

and histologic non-responders, and associated quantitative 

changes in liver fat as measured via MRI-PDFF. Results: Of the 

35 patients who underwent paired liver biopsy and MRI-PDFF 

assessment, 10 demonstrated histologic response. Histologic 

responders and non-responders had similar baseline demographic 

and histological characteristics, with a median age 

of 60.5 years and 70% female vs. median age of 49 years 

and 64% female, respectively. Compared to histologic non-responders, 

histologic responders had a statistically significant 

reduction in net MRI-PDFF of -4.1% ± 4.9 vs. +0.6% ± 4.1 (P 

< 0.036) with a mean percent change of -29.3% ± 33.0 vs. 

+2.0% ± 24.0 (P < 0.004), respectively. Histologic responders 

had a significant decrease in hepatic steatosis (P = 0.007) and 

hepatocellular ballooning (P = 0.025) compared to histologic 

non-responders, and no significant changes were seen in lobular 

inflammation and fibrosis scores within or between groups. 

Conclusion: Utilizing paired MRI-PDFF and liver histology data 

from MOZART Trial, we demonstrate that a 29% reduction in 

liver fat on MRI-PDFF is associated with histologic response in 

NASH. These novel data can be incorporated into designing 

future NASH clinical trials, especially those utilizing change in 

liver fat quantified by MRI as an end-point.


Disclosures: 

Lisa M. Richards - Advisory Committees or Review Panels: Merck; Speaking and 

Teaching: AbbVie, BMS, Gilead 







The following authors have nothing to disclose: Janki R. Patel, Ricki Bettencourt, 

Jeffrey Y. Cui, Joanie Salotti, Jonathan Hooker, Archana Bhatt, Carolyn Hernandez, 

Phirum Nguyen, Hamed Aryafar, William Haufe, Catherine A. Hooker 

914 

Mast Cell Involvement in Non-Alcoholic Fatty Liver Disease 

in Humans 

Anna Christina Dela Cruz 1 , Mehmet M. Altintas 1 , Nestor De La 

Cruz-Munoz 2 , Gabriel S. Gaidosh 4 , Leopoldo Arosemena 1 , Mehrdad 

Nadji 3 , Monica Garcia-Buitrago 3 , Ali Nayer 1 ; 1 Department of 

Medicine, University of Miami Miller School of Medicine, Miami, 

FL; 2 Department of Surgery, University of Miami Miller School 

of Medicine, Miami, FL; 3 Department of Pathology, University of 

Miami Miller School of Medicine, Miami, FL; 4 Bascom Palmer Eye 

Institute, University of Miami Miller School of Medicine, Miami, FL 

Inflammation plays a critical role in the progression of non-alcoholic 

fatty liver disease (NAFLD). Mast cells accumulate in 

the liver of rodents with NAFLD and may contribute to the 

progression of liver disease. It is not known whether mast cells 

are involved in human NAFLD. We examined liver biopsies 

from humans with NAFLD (n=42) and controls (n=7). Hepatic 

steatosis, necroinflammatory activity, and fibrosis were graded 

according to the Batts-Ludwig classification. Mast cells were 

identified using c-Kit and tryptase immunostaining. Two pathologists 

independently determined the density of c-Kit + mast cells 

in the liver in a blinded fashion. To determine the effect of mast 

cell deficiency on the development of NAFLD, 6-week-old male 

Kit W /Kit W-v and congenic control mice were fed a high-fat diet 

(60% energy from fat) for 18 weeks. Data was expressed in 

mean±SEM. Student’s t-test and Pearson’s test were used to 

analyze the data (GraphPad Prism, version 5.0a). P values of 

less than 0.05 were considered statistically significant. There 

was significant inter-observer correlation with respect to mast 

cell quantification (r=0.78, p


916 

An Intact Fgf15 Signaling Pathway is Necessary for 

Hepatoprotection after Bariatric Surgery 

Andriy Myronovych 1,2 , Kenneth D. Setchell 3 , Brandon Tan 4 , 

Rosa-Maria Salazar-Gonzalez 2 , Wujuan Zhang 3 , Karen K. Ryan 5 , 

Darleen A. Sandoval 1 , Randy J. Seeley 1 , Rohit Kohli 2 ; 1 Surgery, 

University of Michigan, Ann Arbor, MI; 2 Gastroenterology, Hepatology 

and Nutrition, Cincinnati Children’s Hospital Medical 

Center, Cincinnati, OH; 3 Pathology and Laboratory Medicine, 

Cincinnati Children’s Hospital Medical Center, Cincinnati, OH; 

4 College of Medicine, University of Cincinnati, Cincinnati, OH; 

5 Department of Physiology & Membrane Biology, University of 

California, Davis, Davis, CA 

Introduction: Vertical sleeve gastrectomy (VSG) results in the 

reduction of body weight and nonalcoholic fatty liver disease 

(NAFLD). Serum bile acids, well-recognized signaling molecules, 

are increased after VSG and their nuclear receptor, 

farnesoid X receptor (Fxr) plays a role in regulating metabolic 

improvements seen following VSG. Intestinal fibroblast growth 

factor 15 (Fgf15) is the Fxr target which suppresses liver lipogenesis. 

We hypothesized that an intact Fgf15 is necessary for 

NAFLD resolution and hepatoprotection after VSG. Methods: 

Fgf15 knockout mice (Fgf15 KO) and their wild type (WT) 

littermates were fed a high fat diet (HFD, 60 kcal%) to induce 

obesity. Mice were randomized into groups, viz.; VSG surgery 

on Fgf15 KO (Fgf15 KO VSG) and WT (WT VSG) mice, Sham 

surgery on Fgf15 KO (Fgf15 KO Sham) and WT (WT Sham) 

mice. Animals received the same HFD thereafter. Mice were 

sacrificed 7 weeks post-surgery. Results: Mice post-VSG lost 

more weight than their respective Sham groups (p


918 

Wheat Bran Autolytic Peptides, Containing a Branchedchain 

Amino Acid, Ameliorate Nonalcoholic Steatohepatitis 

with Up-regulation of Anti-oxidant Potential in 

High-fat Diet-fed Mice 

Takumi Kawaguchi 4 , Takato Ueno 1,3 , Yoichi Nogata 2 , Masako 

Hayakawa 3 , Hironori Koga 4,3 , Takuji Torimura 4,3 ; 1 Asakura 

Medical Association Hospital, Asakura, Japan; 2 NARO Western 

Region Agricultural Research Center, Kagawa, Japan; 3 Liver 

Cancer Division, Research Center for Innovative Cancer Therapy, 

Kurume University, Kurume, Japan; 4 Division of Gastroenterology, 

Department of Medicine, Kurume University School of Medicine, 

Kurume, Japan 

Backgrounds and Aims: Intake of whole wheat is known to 

decrease risk of lifestyle-related diseases. Recently, we have 

developed a simple extracting method of antihypertensive peptides 

from wheat bran by autolysis reactions. The wheat bran 

autolytic peptides (WBAPs) contain leucine, a branched-chain 

amino acid, which ameliorates oxidative stress and insulin 

resistance. The aims of this study are to investigate therapeutic 

efficacy of WBAPs for nonalcoholic steatohepatitis (NASH) in 

a mouse model. In addition, we evaluate effects of WBAPs 

on oxidative stress and anti-oxidant potential. Material and 

Methods: Seven-week-old male C57BL/6 mice were fed with a 

high fat diet for 10 weeks to induce NASH, and were simultaneously 

administrated with either distilled water (CON; n=5), 

or WBAP-1 supplemented water (0.05% leucine-arginine-proline; 

n=5), or WBAP-2 supplemented water (0.05% leucine-glutamine-proline; 

n=5) ad libitum. Then, they were sacrificed, 

and body weight, nonalcoholic fatty liver disease activity score 

(NAS), and its components were evaluated. Moreover, severity 

of oxidative stress in liver tissues was evaluated by measurement 

of derivate of reactive oxygen metabolites (d-ROM) and 

biological antioxidant potential (BAP) using free radical elective 

evaluator (FREE cape diem, DIACRON INTERNATIONAL 

s.r.l.). Results: No significant difference was seen in body 

weight between the CON and WBAP-1 groups. While, body 

weight in the WBAP-2 group was significantly lower compared 

to the CON group (46.0±3.0 vs. 38.0±3.3 g/body, P=0.02). 

The grades of steatosis, lobular inflammation, and hepatocyte 

ballooning was significantly lower in the WBAP-1 and WBAP-2 

groups compared to the CON group. NAS was also significantly 

lower in the WBAP-1 and WBAP-2 groups compared to 

the CON group (CON; 6.6±0.5, WBAP-1; 1.8±2.0, WBAP- 

2; 3.8±2.4, P=0.02). There was no significant difference in 

hepatic d-ROM levels among the groups. Although no significant 

difference was seen in hepatic BAP levels between the 

CON and WABP-2 groups, hepatic BAP levels in the WBAP-1 

group were significantly higher compared to the CON group 

(3242± 338 vs. 2549±63 mmol/L, P=0.03). Conclusions: We 

demonstrated that WBAPs ameliorated NASH in a high-fat 

diet-induced mouse model. Furthermore, WBAPs up-regulated 

BAP. Since WBAPs can be obtained easily at a low cost, 

WBAPs may be clinically applicable agents in patients with 

NASH. 

Disclosures: 

The following authors have nothing to disclose: Takumi Kawaguchi, Takato Ueno, 

Yoichi Nogata, Masako Hayakawa, Hironori Koga, Takuji Torimura 

919 

Hepatic but not Intestinal Specific FXR Deficiency Led to 

Accelerated Non-alcoholic Steatohepatitis Development 

Bo Kong 1 , Runbin Sun 1,2 , Jianliang Shen 1 , Yang Pan 1 , Jia He 1 , 

Quan Jin 1 , Justin D. Schumacher 1 , Le Zhan 1 , Haoyue Zhou 1 , Xinjie 

Qiu 1 , Yongping Wang 3 , Jiye Aa 2 , Jason R. Richardson 4 , Tracy 

Anthony 3 , Grace L. Guo 1 ; 1 Department of Pharmacology and Toxicology, 

Rutgers University, Piscataway, NJ; 2 Key Laboratory of 

Drug Metabolism and Pharmacokinetics, China Pharmaceutical 

University, Nanjing, China; 3 Department of Nutritional Sciences, 

Rutgers University, New Brunswick, NJ; 4 Department of Environmental 

& Occupational Medicine, Robert Wood Johnson Medical 

School, Piscataway, NJ 

Non-alcoholic fatty liver disease (NAFLD) is a spectrum of 

chronic liver diseases ranging from simple hepatic steatosis to 

non-alcoholic steatohepatitis (NASH), characterized by steatosis 

with inflammation. NAFLD can progress to fibrosis, cirrhosis, 

and liver tumor. The mechanisms underlying the transition from 

simple steatosis to NASH are still elusive and effective therapies 

to treat NASH are unavailable. Our previous study showed 

that FXR deficiency and/or altered bile acid homeostasis is 

involved in the development of NASH in mice, and FXR ligands 

emerge as novel treatment strategy for NASH in humans. However, 

the tissue specific roles of FXR in NASH development 

are not clear. In this study, wild-type (WT; C57BL/6J), wholebody 

FXR knockout (FXR KO), hepatocyte-specific FXR knockout 

(FXR Hep-/- ) and enterocyte-specific FXR knockout (FXR Int-/- ) mice 

were fed a high-fat diet (HFD) with 60% calories from fat for six 

months. Glucose tolerance test and serum liver functional tests, 

as well as body weight changes and lipid parameters were 

measured for monitoring metabolic syndrome development. 

H&E and Sirius red staining were performed to determine the 

degree of pathological changes and fibrosis. Hepatic levels of 

triglycerides and cholesterols were determined to assess the 

degree of liver steatosis. Changes in the expression of genes 

involved in lipid metabolism, bile acid homeostasis, inflammation, 

fibrogenesis and ER stress were determined using Q-PCR. 

Consistent with our previous report, the results showed that HFD 

fed WT mice developed steatosis, inflammation, and fibrosis, 

resembling human NASH characteristics. In detail, they had 

increased body weight, reduced insulin sensitivity, increased 

serum and hepatic levels of triglycerides, cholesterols and alanine 

aminotransferase (ALT) activities, enhanced expression 

of inflammation-related genes (IL-6, TNFα, ICAM-1), ER stress 

(Chop) and fibrogenesis-related genes (Collagen 1a1, TIMP-1). 

FXR KO mice showed exacerbated phenotype compared to 

WT mice on HFD. Furthermore, tissue-specific deletion of the 

FXR gene affects NASH severity. Specifically, on HFD FXR Hep-/- 

mice manifested severe NASH phenotype comparable to that 

in FXR KO mice, but the FXR Int-/- mice showed similar changes 

to WT mice. In conclusion, FXR in hepatocytes plays a critical 

role in the protection against NASH development. The study 

suggests that activation of FXR in hepatocytes may represent a 

better strategy for NASH prevention and treatment. 

Disclosures: 

Grace L. Guo - Consulting: NGM 

The following authors have nothing to disclose: Bo Kong, Runbin Sun, Jianliang 

Shen, Yang Pan, Jia He, Quan Jin, Justin D. Schumacher, Le Zhan, Haoyue Zhou, 

Xinjie Qiu, Yongping Wang, Jiye Aa, Jason R. Richardson, Tracy Anthony


920 

Deletion of orphan nuclear receptor small heterodimer 

partner prevents development of nonalcoholic steatohepatitis 

Chunki Kim, Mikang Lee, Yoonkwang Lee; Integrative Medical 

Sciences, Northeast Ohio Medical University, Rootstown, OH 

An orphan nuclear hormone receptor Small Heterodimer 

Partner (SHP) is involved in lipid and bile acid metabolism. 

Our earlier studies have shown that deletion of SHP protects 

the mice from diet-induced obesity and hepatic steatosis. In 

order to explore a potential role of SHP in the development 

of NASH, we quantified inflammatory gene expression in the 

mice fed a westernized diet (WestD) for 6 month. The longterm 

diet regimen induced mRNA levels of inflammatory genes 

such as TNFa, IL-1b, CXCL-1, and iNOS in wild type (WT) 

mice. However, the expression of these genes was significantly 

down-regulated in SHPKO counterparts. In order to find out 

whether the reduction of inflammatory responses is due to 

SHP gene deletion or lower hepatic lipid level, we generated 

ApoE/SHP double knockout mice (ApoE/SHPKO). ApoEKO 

mouse model has been shown to be susceptible to diet-induced 

inflammation but resistant to hepatic steatosis. One week- 

WestD regimen significantly induced the expression of the 

pro-inflammatory genes in ApoEKO mice but not in ApoE/ 

SHPKO mice without evidence of hepatic TG accumulation in 

either genotype as expected, suggesting SHP, not hepatic TG 

level, is responsible for the observed inflammatory responses. 

To study NASH development in SHPKO mice further, the mice 

were challenged with methionine and choline-deficient (MCD) 

diet for 6 weeks. SHPKO mice were strongly protected from 

NASH development evidenced by reduced pro-inflammatory 

gene expression, reduced F4/80 positive macrophage infiltration, 

decreased mRNA levels of fibrosis-related genes such 

as collagen 1a1, TGF-b1, TIMP1, and a-SMA, which were all 

significantly induced in WT mice. In addition, hepatocyte specific-SHPKO 

(AlbCre/floxedSHP) mice challenged with MCD 

diet also showed significant protection in NASH development, 

as assessed with qPCR, immunofluorescence, Sirius red staining, 

compared to floxed SHP counterparts. The current study 

identifies the role of SHP as pro-inflammatory in the liver and 

will provide new insights into development of NASH and its 

possible therapeutic or preventative approaches. 

Disclosures: 

The following authors have nothing to disclose: Chunki Kim, Mikang Lee, Yoonkwang 

Lee 

921 

Differential carbonylation of proteins in human fatty 

and nonfatty NASH 

Colin T. Shearn, David J. Orlicky, Dennis R. Petersen; Department 

of Pathology, University of Colorado Anschutz Medical Campus, 

Aurora, CO 

Objective: In the liver, a contributing factor in the pathogenesis 

of non-alcoholic fatty liver disease is oxidative stress leading 

to the accumulation of highly reactive electrophilic α/β unsaturated 

aldehydes. The objective of this study was to determine 

if significant differences were evident when evaluating carbonylation 

in human end-stage fatty nonalcoholic steatohepatitis 

(fNASH) compared to end stage non-fatty NASH (nfNASH). 

Methods: Using age-matched pooled hepatic tissue obtained 

from healthy humans and patients diagnosed with end stage 

nfNASH or fNASH, overall carbonylation was assessed by 

immunohistochemistry (IHC) and LC-MS/MS of streptavidin 

purified hepatic whole cell extracts treated with biotin hydrazide. 

Identified carbonylated proteins were further evaluated 

using bioinformatics analyses. Results: Picrosirius red staining 

revealed extensive fibrosis in both fNASH and nfNASH which 

corresponded with increased 4-HNE staining. Although significantly 

elevated when compared to normal hepatic tissue, 

no significant differences in overall carbonylation and fibrosis 

were evident when comparing fNASH with nfNASH. Mass 

spectrometric analysis revealed a total of 184 carbonylated 

proteins. Of these, 51 were unique to nfNASH, 32 unique to 

fNASH, 33 unique to normal hepatic tissue and 27 common to 

all groups. DAVID and Gene Ontology bioinformatic pathway 

analysis of hepatic carbonylated proteins revealed a propensity 

for increased carbonylation of carboxylic acid catabolic 

process, generation of precursor metabolites as well as energy 

and proteolysis in nfNASH. Whereas carbonylation of proteins 

implicated in protein complex biogenesis and regulation of 

apoptosis occurred in fNASH. Using LC-MS/MS analysis, in 

nfNASH a nonenaldehyde adduct was identified on Lys 235 

on the cytoskeletal protein vimentin. Conclusions: These results 

suggest that cellular factors regulating mechanisms of protein 

carbonylation may be different depending on pathological 

diagnosis of NASH. Furthermore these studies are the first to 

use LC-MS/MS analysis of carbonylated proteins in human 

NAFLD and begin exploring possible mechanistic links with 

end stage cirrhosis due to fatty liver disease and oxidative 

stress. 

Disclosures: 

The following authors have nothing to disclose: Colin T. Shearn, David J. Orlicky, 

Dennis R. Petersen 

922 

Lysosomal cholesterol in Kupffer cells, particularly when 

oxidized, contributes to murine steatohepatitis 

Sofie Walenbergh 1 , Tom Houben 1 , Tim Hendrikx 1 , Patrick van 

Gorp 1 , Mike Jeurissen 1 , Marie-Hélène Lenders 1 , Marion J. Gijbels 

1 , Jogchum Plat 1 , Marten H. Hofker 2 , Christoph J. Binder 3 , 

Dieter Luetjohann 4 , Fons Verheyen 1 , Ger H. Koek 1 , Ronit Shiri-Sverdlov 

1 ; 1 Maastricht University, Maastricht, Netherlands; 

2 University Medical Center Groningen, Groningen, Netherlands; 

3 Research Center for Molecular Medicine of the Austrian Academy 

of Sciences, Vienna, Austria; 4 Institute of Clinical Chemistry and 

Clinical Pharmacology, Bonn, Germany 

Background and Aims: Recently, the importance of lysosomes 

within the metabolic syndrome, including fatty liver disease, 

is gaining increasing attention. It has been suggested that 

macrophages during atherosclerosis as well as Kupffer cells 

(KCs) during hepatic inflammation demonstrate properties of 

an acquired lysosomal storage disorder. So far, it is unclear 

whether there is a causal relationship between lysosomal cholesterol 

accumulation (LCA) in KCs and hepatic inflammation. 

Additionally, the specific contribution of the oxidized LDL 

(oxLDL) fraction to LCA, its concomitant effect on lysosomal 

function and on hepatic inflammation is unexplored. Methods: 

Hematopoietic deficiency of the mutant Niemann-Pick type C1 

(NPC1 mutant ) protein was used as a tool to induce LCA in KCs 

of hyperlipidemic mice. To induce high levels of anti-oxLDL antibodies, 

NPC1 mutant -transplanted mice were immunized every 

two weeks with heat-inactivated pneumococci until the end 

of the experiment. Results: Compared to wildtype, NPC1 mutant 

-transplanted mice displayed severe hepatic inflammation 

and fibrosis. Anti-oxLDL immunization of NPC1 mutant -transplanted 

mice improved cholesterol metabolism, lysosomal dysfunction 

and liver inflammation, in contrast to non-immunized 

NPC1 mutant -transplanted mice. Conclusions: A direct causal link 

exists between LCA in KCs and hepatic inflammation. Rather


than total cholesterol, we specifically show that oxLDL significantly 

contributes to lysosomal dysfunction, cholesterol homeostasis 

and the hepatic inflammatory response. 

Disclosures: 

The following authors have nothing to disclose: Sofie Walenbergh, Tom Houben, 

Tim Hendrikx, Patrick van Gorp, Mike Jeurissen, Marie-Hélène Lenders, Marion J. 

Gijbels, Jogchum Plat, Marten H. Hofker, Christoph J. Binder, Dieter Luetjohann, 

Fons Verheyen, Ger H. Koek, Ronit Shiri-Sverdlov 

923 

Cholesterol crystallization within hepatocyte lipid droplets 

and its role in NASH 

George N. Ioannou 2,1 , Savitha Subramanian 1 , Alan Chait 1 , Matthew 

M. Yeh 1 , W. G. Haigh 2 , Christopher Savard 2,1 ; 1 University 

of Washington, Seattle, WA; 2 Veterans Affairs Puget Sound 

Healthcare System, Seattle, WA 

Background/Aims We recently reported that cholesterol crystals 

were present within the lipid droplets of hepatocytes in 

patients and mouse models with NASH. We sought to further 

characterize the process of cholesterol crystallization in hepatocyte 

lipid droplets and its role in the development of Kupffer 

cell (KC) crown-like structures (CLS). Methods C57BL/6J, wildtype 

mice were assigned to a high-fat (15%, w/w) diet for 

6 months supplemented with 0%, 0.25%, 0.5%, 0.75%, or 

1% dietary cholesterol (5 groups, n=12 mice/group). HepG2 

hepatoma cells were exposed to varying concentrations of LDL 

cholesterol, oleic acid, and ACAT inhibitor, in order to induce 

lipid droplet formation and cholesterol crystallization. Results 

Fibrosing steatohepatitis developed at a dietary cholesterol 

concentration ≥0.5%, whereas mice on a diet of 0% or 0.25% 

cholesterol developed only simple steatosis. Hepatic cholesterol 

crystals and CLSs were also only observed at a dietary cholesterol 

concentration ≥0.5%. CLSs consisted of activated KCs that 

surrounded and processed cholesterol-crystal containing remnant 

lipid droplets of dead hepatocytes and stained intensely 

positive for NLRP3 and activated caspase1. When HepG2 

cells were exposed to 2000 mg/ml LDL and 200 mM Oleic 

acid for >20 days, a sub-population of cells displayed birefringent 

cholesterol crystals around the periphery of large lipid 

droplets. Conclusion A specific threshold dietary cholesterol 

concentration that leads to cholesterol crystallization within 

hepatocyte lipid droplets also leads to CLSs and fibrosing 

NASH, suggesting a causative association. Exposure of KCs in 

CLSs to cholesterol crystals activates the NLRP3 inflammasome. 

We developed an in vitro cell culture model of steatosis and 

cholesterol crystallization in HepG2 cells. 

Disclosures: 

The following authors have nothing to disclose: George N. Ioannou, Savitha 

Subramanian, Alan Chait, Matthew M. Yeh, W. G. Haigh, Christopher Savard 

924 

Fenofibrate and LXRα agonist combination attenuated 

intrahepatic inflammation in non-alcoholic fatty liver. 

Eun Chul Jang 1 , Dae Won Jun 2 , Seung Min Lee 3 , Yong Kyun 

Cho 4 , Sang Bong Ahn 5 ; 1 Department of Occupational and Environmental 

Medicine, Soonchunhyang University Cheonan Hospital, 

Cheonan, Korea (the Republic of); 2 Department of Internal 

Medicine, Hanyang University College of Medicine, Seoul, Korea 

(the Republic of); 3 Department of Food and Nutrition, Sungshin 

Women’s University, Seoul, Korea (the Republic of); 4 Department 

of Internal Medicine, Kangbuk Samsung Hospital, Sungkyunkwan 

University, School of Medicine, Seoul, Korea (the Republic of); 

5 Department of Internal Medicine, Eulji University School of Medicine, 


Background/Aims: Liver X receptors (LXR) is key transcription 

factor in the regulation of lipid and cholesterol metabolism. 

in addition, LXRα has been implicated as regulator of inflammation. 

LXRα activation is associated with hepatic steatosis 

and hyperlipidemia in mice. Fenofibrate, a PPARα agonist and 

omega-3, an antihypertriglyceride agent, lead to a reduction 

of hyperlipidemia. The aim of this study to investigate whether 

concurrent effect of LXRα and fenofibrate or omega-3 can produce 

synergic benefits in high-fat diet induced obese mice. 

Methods: Normal chow and high-fat diet mice treated with 

LXRα agonist (T0901317), or combined fenofibrate or omega-3 

for 4 weeks. Hematoxylin and eosin staining was performed 

on liver tissue extracts after animal sacrifice. SREBP1c, FAS, 

SCD-1, PPAR-α, and MCP-1 mRNA expressions were assessed 

with reverse transcription-polymerase chain reaction. Results: 

LXRα agonist increase intrahepatic fat amount in normal chow 

group. Degree of intrahepatic inflammation was not different 

among LXRα agonist, or combined with fenofibrate/omega- 

3. In NAFLD model, the combined treatment with fenofibrate 

did not increase hepatic steatosis. LXRα agonist and/or fenofibrate 

or omega-3 decreased intrahepatic inflammation. In 

high-fat diet groups, combined treatment with fenofibrate markedly 

reduced the expression of genes involved in lipogenesis, 

including srebp-1c, fas and scd1. Furthermore, LXRα agonist 

and/or fenofibrate or omega3 treatment decreased expression 

of abca-1, abcg5, and abcg8 genes, three vital genes for 

cholesterol efflux in diet induced fatty liver model, but those 

expressions were opposite in normal chow group. Intraheptic 

MCP-1, and TNF-α expression and markers of inflammasome 

were also decreased in NAFLD group. Conclusion: LXRα agonist 

and fenofibrate combination treatment attenuated hepatic 

inflammation in NAFLD model. 

Disclosures: 

The following authors have nothing to disclose: Eun Chul Jang, Dae Won Jun, 

Seung Min Lee, Yong Kyun Cho, Sang Bong Ahn


925 

A novel transcriptional repressor Hairy and Enhancer 

Split 6 mediates hepatic lipid homeostasis through inhibition 

of Pparg2 expression 

Chunki Kim, Mikang Lee, James P. Hardwick, Yoonkwang Lee; 

IMS, Northeastern Ohio Univ. College of Med, Rootstown, OH 

Hepatic steatosis is a required prelude for development of 

non-alcoholic fatty liver diseases. Pparg is a member of nuclear 

hormone receptor superfamily and a master regulator for white 

adipocyte differentiation and lipid storage. Uprising hepatic 

Pparg2 level reprograms liver for lipid storage instead of for 

VLDL secretion or fatty acid oxidation, which leads to hepatic 

fat accumulation in pathophysiological conditions such as 

obesity and diabetes. Our earlier study suggested that small 

heterodimer partner and retinoic acid receptor coordinately 

regulates Pparg2 gene expression via directly regulating a 

novel transcriptional repressor Hairy and Enhancer Split 6 

(Hes6) mRNA level. In the suggested regulatory paradigm, 

Hes6 represses Pparg2 gene expression by inhibiting DNAbound 

HNF4a transcriptional activity. To explore function of 

Hes6 in hepatic lipid mobilization, we overexpressed Hes6 

using adenovirus in the livers of mice fed a westernized diet 

for 2 months. The targeted overexpression reduced Pparg2 

mRNA level by 60% and hepatic triglyceride accumulation by 

30% compared to the levels obtained from mice injected with 

adenoviral empty vector. We also silenced hepatic Hes6 using 

adenoviral shRNA and fed the mice western diet for additional 

2 weeks. The adenoviral shHes6 effectively silenced Hes6 

gene expression by approximately 90% and increased hepatic 

fat accumulation and Pparg2 mRNA level by 50% and 6 fold, 

respectively. In subsequent experiments, we utilized transient 

transfection and gel mobility shift assays to identify a specific 

HNF4a binding element in the Pparg2 promoter. Indeed, an 

Hnf4a binding consensus sequence was identified at -903bp 

from transcription start site. Deletion or point mutation of the 

sequence in a luciferase reporter containing the Pparg2 promoter 

abolished Hes6-mediated repression in HepG2 cells. 

Chromatin immunoprecipitation and gel mobility shift assays 

further confirmed direct recruitment and binding of Hnf4a to 

the site. The data strongly prove that expression of Pparg2 is 

maintained low by coordinate repression by Hes6 and Hnf4a 

in normal liver and thus the Hes6-Hnf4a-Pparg2 transcriptional 

axis is considered as a critical determinant for hepatic lipid 

homeostasis. 

Disclosures: 

The following authors have nothing to disclose: Chunki Kim, Mikang Lee, James 

P. Hardwick, Yoonkwang Lee 

926 

Effects of Dietary Different Doses of Copper and High 

Fructose Feeding on Rat fecal Metabolome 

Ming Song 1 , Xiaoli Wei 2 , Xinmin Yin 2 , Dale Schuschke 3 , Imhoi 

Koo 2 , Craig J. McClain 1,4 , Xiang Zhang 2 ; 1 Department of Medicine/GI, 

University of Louisville, Louisville, KY; 2 Chemistry, University 

of Louisville, Louisville, KY; 3 Physiology, University of Louisville, 

Louisville, KY; 4 Robley Rex VAMC, Louisville, KY 

Background/Aims: The gut microbiota play a critical role in 

the pathogenesis of nonalcoholic fatty liver disease (NAFLD) 

through altering the gut metabolites and gut barrier function. 

Increased fructose consumption and inadequate copper intake 

are two critical risk factors in the development of NAFLD. The 

aim of this study was to determine the effect of different dietary 

doses of copper and high fructose feeding on fecal metabolites 

in a rat model. Methods: To gain insights into the role of gut 

microbiota, male weanling Sprague-Dawley rats were exposed 

to different dietary levels of copper (adequate copper, 6ppm; 

marginal copper, 1.5ppm; supplemental copper, 20ppm) with 

and without high fructose (30% fructose, w/v, in the drinking 

water was given ad lib) intake for 4 weeks. Fecal metabolites 

of each rat were then analyzed by comprehensive two-dimensional 

gas chromatography time-of-flight mass spectrometry 

(GC×GC-TOF MS). In parallel, liver tissues were assessed by 

histology and triglyceride assay. Results: Our data showed 

that high fructose feeding led to obvious hepatic steatosis in 

both marginal copper deficient rats and copper supplementation 

rats. Among the 38 metabolites detected with significant 

abundance alteration between groups, short chain fatty acids 

(SCFAs) were markedly decreased with excessive fructose 

intake, irrespective of copper levels. C15:0 and C17:0 long 

chain fatty acids (LCFAs), produced only by bacteria, were 

increased by either high copper level or high fructose intake. 

In addition, increased fecal urea and malic acid paralleled 

the increased hepatic fat accumulation. Conclusion: GC×GC- 

TOF MS analysis of rat fecal samples revealed distinct fecal 

metabolome profiles associated with the dietary high fructose 

and copper level, with some metabolites possibly serving as 

potential noninvasive biomarkers of fructose induced-NAFLD. 

Disclosures: 




The following authors have nothing to disclose: Ming Song, Xiaoli Wei, Xinmin 

Yin, Dale Schuschke, Imhoi Koo, Xiang Zhang 

927 

Liver sinusoid endothelial cell derived bone morphogenetic 

protein binding endothelial regulator (BMPER) 

induces iron overload of high-fat diet induced non-alcoholic 

fatty liver mice 

Takumu Hasebe 1 , Koji Sawada 1 , Shunsuke Nakajima 1 , Hiroki 

Tanaka 2 , Takaaki Ohtake 3 , Mikihiro Fujiya 1 , Yutaka Kohgo 3 ; 

1 Medicine, Asahikawa Medical University, Asahikawa, Japan; 

2 Gastrointestinal Immunology and Regenerative Medicine, Asahikawa 

Medical University, Asahikawa, Japan; 3 Gastroenterology, 

International University of Health and Welfare Hospital, 

Otawara, Japan 

[Background] Excessive irons frequently coexist with non-alcoholic 

fatty liver (NAFL), which induces hepatic inflammation 

and fibrosis. Down regulation of iron regulatory protein 

hepcidin and its inducer bone morphogenetic protein (BMP) 

signals can be the central cause of iron overload, however 

the mechanism in NAFL is still controversial. To investigate 

the mechanism of iron overload in NAFL, we performed transcriptome 

analysis using high throughput sequencer. [Methods] 

Male C57BL/6 mice were fed on regular or high-fat diet for 16 

weeks. Internal iron was evaluated by plasma iron, ferritin or 

hepatic iron content. Whole RNA sequencing was performed 

as transcriptome analysis using Ion Proton (Life Technologies). 

Altered expressions of genes comparing between regular and 

high-fat diet mice were as follows; fold change > 1.5, p value 

< 0.05 (Student’s t test). Altered gene expressions in mice liver 

were confirmed by RT-PCR. Plasma hepcidin concentration was 

measured by LC-MS/MS. Localization of protein expression 

in mice liver was analyzed by immunofluorescence. Hepatocytes 

and liver sinusoid endothelial cells were isolated from 

regular mice by collagenase perfusion to assess expressions 

of iron regulating molecule by RT-PCR. [Results] High-fat diet 

mice showed significant obesity and fatty liver, however neither 

hepatic inflammation nor fibrosis. Plasma iron and ferritin were 

increased in high-fat diet mice, whereas hepatic iron content


was not changed. RNA sequencing showed 2314 expression 

altered genes of total 38114 analyzed. When we focus on 

iron regulatory proteins, we found decreased hepcidin and 

increased BMP-SMAD signal related genes: Bmp4, Bmper and 

Hfe2. In contrast, the other hepcidin inducing signal: transferrin 

receptor signal and IL-6 signal, related genes were not 

altered. Plasma hepcidin concentration in high-fat mice was 

decreased and the hepcidin inducer SMAD phosphorylation 

was decreased. BMP binding endothelial regulator (BMPER), a 

BMP inhibiting protein, is increased in high-fat diet mice. Immunofluorescense 

of frozen liver section showed BMPER expression 

localized on sinusoid lumen. Cell isolation also showed 

higher expression of BMPER from liver sinusoid endothelial 

cells. [Discussions] Excessive irons in high-fat diet fed mice 

were caused by decreased hepcidin. We identified that BMPER 

is down-regulating hepcidin via BMP-SMAD signaling inhibition. 

We also discovered BMPER is expressed by liver sinusoid 

endothelial cells to work on hepcidin producer hepatocyte 

extracellularly. This study shows importance of interaction with 

hepatocytes and non-parenchymal cells in NAFL pathogenesis. 

Disclosures: 

Yutaka Kohgo - Grant/Research Support: Chugai Pharm., Novartis Japan, Asahikasei 

Medical, Sapporo Beer Co. 

The following authors have nothing to disclose: Takumu Hasebe, Koji Sawada, 

Shunsuke Nakajima, Hiroki Tanaka, Takaaki Ohtake, Mikihiro Fujiya 

928 

Analysis of T and myeloid cells in patients with NASH 

and diabetes mellitus 

Luisa Vonghia 1 , Denis Mogilenko 2,3 , An Verrijken 4,5 , Luc van 

Gaal 4,5 , Bart Staels 2,3 , Sven M. Francque 1,5 , David Dombrowicz 

2,3 ; 1 Department of Gastroenterology and hepatology, University 

Hospital, Antwerp, Belgium; 2 Inserm U1011, Lille, France; 

3 Institut Pasteur, University of Lille, Lille, France; 4 Department of 

Endocrinology, Diabetology and Metabolism, University Hospital 

of Antwerp, Antwerp, Belgium; 5 Laboratory of Experimental Medicine 

and Paediatrics, Faculty of Medicine and Health Sciences, 

University of Antwerp, Antwerp, Belgium 

BACKGROUND: The immune system potentially plays a pivotal 

role in the onset of Non-Alcoholic Steatohepatitis (NASH) and 

of the associated metabolic disturbances, including diabetes 

mellitus (DM). AIM: To study the immune cells in peripheral 

blood and differentially expressed genes in the liver of preselected 

patients according to presence or absence of NASH and 

DM. METHODS: We enrolled 32 patients who underwent liver 

biopsy because of suspected NASH. The patients were divided 

in 4 predefined groups according to liver biopsy and glucose 

parameters: 1) control (NO NASH/NO DM), 2) NASH/NO 

DM, 3) NASH/DM, 4) NO NASH/DM. A multicolour flow 

cytometry analysis was performed in order to investigate T, 

natural killer T cells (NKT), natural killer (NK) cells, monocytes 

and dendritic cells. Gene expression in the liver was investigated 

by microarray analysis in patients with similar clinical 

characteristics. RESULTS: Significant differences between 

groups of patients were found among T lymphocyte populations 

but not in subsets of myeloid cells. Proportions of CD8 + 

cells with enhanced production of pro-inflammatory cytokines 

(TNF and INFγ) and cytotoxic molecules (granzyme A and B, 

perforin) were increased in patients with NASH and/or DM. 

The central memory CD8 + cells (CCR7 + CD45RA - ) were significantly 

increased in NASH/DM compared to NASH/NO DM, 

whereas the effector (CCR7 - CD45RA + ) and effector memory 

(CCR7 - CD45RA - ) CD8 + cells were more potent to produce cytotoxic 

molecules in all groups of patients with NASH and DM. 

Gene set enrichment analysis evenso revealed dysregulation of 

genes associated with activation of CD8 + cells in the liver in 

NASH/DM compared to NASH/NO DM. We found a trend 

to increased proportion of NKT cells in patients with NASH. 

CD4 + T regulatory (Treg) and Th22 cells were increased in 

NASH/DM compared to NASH/NO DM patients, while Tregs 

where decreased in NASH patients without DM. CONCLU- 

SION: T-cell, bot not myeloid cell, alterations are associated 

with the presence of NASH regardless of the presence of diabetes. 

Both flow cytometry and gene expression demonstrate 

that increased effector functions of cytotoxic CD8 + T cells and 

CD4 + Th22 and Treg cells are found when NASH is associated 

with DM, suggesting that these cells are specifically involved 

in the crosstalk between liver inflammation and metabolic dysfunctions 

and contribute both to hepatotoxic and hepatoprotective 

mechanisms. 

Disclosures: 


The following authors have nothing to disclose: Luisa Vonghia, Denis Mogilenko, 

An Verrijken, Luc van Gaal, Sven M. Francque, David Dombrowicz 

929 

Regulation of hepatocellular senescence by melatonin 

during alcoholic liver injury 

Jessica S. Garner 1 , Yuyan Han 3,2 , Tiaohao Zhou 3,2 , Kelly McDaniel 

2,3 , Ying Wan 1,2 , Tami Annable 2,1 , Nan Wu 3,2 , Julie Venter 3,2 , 

Haibo Bai 1,2 , Shannon S. Glaser 3,2 , Heather L. Francis 1,2 , Fanyin 

Meng 2,1 , Gianfranco Alpini 3,2 ; 1 Scott & White Hospital, Texas 

A&M HSC College of Medicine, Temple, TX; 2 Central Texas Veterans 

Healthcare System, Temple, TX; 3 Texas A&M HSC College of 

Medicine, Temple, TX 

Background: Chronic alcohol consumption leads to hepatic 

DNA damage, cellular senescence and permanent cell cycle 

arrest. Melatonin, an endogenously produced neurohormone 

secreted by the pineal gland as well as the liver, has a variety 

of protective effects during organ injury. However, its effect 

and mechanism on the hepatic tissues and cells during alcoholic 

liver injury remains to be explored. The objective of this 

study was to evaluate the role of melatonin regulated hepatic 

senescence phenotype during alcoholic liver injury. Methods: 

The mRNA expression of melatonin, its downstream miRNA 

(miR-34a) and its upstream enzyme serotonin N-acetyltransferase 

(AANAT), was assessed in ethanol and LPS-treated 

human hepatocytes (N-Heps), as well as in 5 weeks chronic 

alcohol feeding mouse liver specimen (with or without antimiR-34a 

Vivo-Morpholino treatment) and control liver tissue 

by PCR array and real-time PCR assay. The secretion of melatonin 

was verified by ELISA assay. Cellular senescence and 

proliferation was evaluated by β-gal activity and MTS assays. 

The hepatic expressions of the senescence genes (EGR1, PAI-1 

and CCL2), clock circadian genes (PER1, BMAL1 and CRY1), 

and miR-34a were also determined by real-time PCR assay. 

Results: The total liver histopathology score, beta-gal activity 

and miR-34a expression increased after 5 weeks chronic ethanol 

feeding relative to control mice, along with the significant 

reduction of melatonin and AANAT in isolated liver tissues. 

Enhanced expression of the senescence markers EGR1, PAI-1 

and CCL2 was demonstrated by senescence PCR array in vivo. 

Treatment with ethanol (20 mM) and LPS (20 μg/ml) for 7 

days induced significant increases of beta-gal staining, along 

with the enhanced expression of cellular senescence markers 

EGR1, PAI-1 and CCL2 in cultured human hepatocytes. Treatment 

of N-Heps with melatonin (10 -11 M for 7 days) also prevented 

alcohol-induced cellular senescence and death, and 

subsequently reduced senescence markers EGR1 and CCL2, 

as well as miR-34a expression. Furthermore, the expression


of melatonin regulated senescence and clock genes, including 

EGR1, PAI-1, CCL2, CLOCK, PER1, BMAL1 and CRY1, 

was significantly altered in chronic ethanol feeding mice livers 

after anti-miR-34a Morpholino treatment relative to the controls. 

Conclusion: The finding that melatonin plays a significant 

role in the regulation of hepatocellular senescence phenotype 

provides the basis for an exciting field in which the melatonin 

related upstream enzyme (AANAT), the downstream miRNAs 

(miR-34a) and clock circadian genes may be manipulated with 

potential therapeutic benefits for alcoholic liver injury. 

Disclosures: 

The following authors have nothing to disclose: Jessica S. Garner, Yuyan Han, 

Tiaohao Zhou, Kelly McDaniel, Ying Wan, Tami Annable, Nan Wu, Julie Venter, 

Haibo Bai, Shannon S. Glaser, Heather L. Francis, Fanyin Meng, Gianfranco 

Alpini 

930 

Hypoxia accelerates fatty acid uptake leading to 

increased fat accumulation and inflammation in mice 

and in cultured human hepatocyte-derived cells 

Agueda Gonzalez-Rodriguez, Gloria Mateo, Ines Soro-Arnaiz, 

Mar Torres-Capelli, Ainara Elorza, Julian Aragones, Carmelo 

García-Monzón; Liver Research Unit, University Hospital Santa 

Cristina, Madrid, Spain 

Nonalcoholic fatty liver disease (NAFLD) is the most common 

cause of chronic liver disease in Western countries. NAFLD is 

strongly associated with overweight/obesity, insulin resistance, 

type 2 diabetes (T2DM) and cardiovascular complications; 

therefore, it is considered the hepatic component of the metabolic 

syndrome. NAFLD is characterized by the progression 

from a benign steatosis to more severe liver injuries directly 

associated with lipotoxicity such as nonalcoholic steatohepatitis 

(NASH), cirrhosis and hepatocellular carcinoma in 10% of 

NAFLD patients. Recent evidence indicates that NAFLD severity 

is affected by obstructive sleep apnoea syndrome (OSAS), a 

recurrent upper-airway obstruction during sleep, leading to 

periods of chronic intermittent hypoxia (CIH). In this regard, 

dysregulation of the normal oxygen gradient in the liver that 

promotes the stabilization of the hypoxia-inducible factors 

(HIFs) can induce liver steatosis and inflammation. However, 

the pathogenic mechanisms underlying the progression of 

NAFLD to NASH in the context of CIH featuring OSAS are not 

fully understood. As working hypothesis, the more pronounced 

the CIH is, the more pronounced the progression to NASH and 

fibrosis. AIM: The purpose of this study was focused on the 

molecular mechanisms linking hypoxia to NAFLD/NASH setup. 

METHODS: HIF system, CD36 content and liver damage markers 

were analyzed in livers from conditional Von Hippel-Lindau 

knockout (VHL-KO) mice, which display an overexpression of 

HIFs after VHL gene deletion induced by tamoxifen, and in 

HepG2 human hepatocytes loaded with palmitic acid submitted 

to an hypoxic environment (1% O 2 

). RESULTS: As expected, 

HIF1 and HIF2 were overexpressed in livers from VHL-KO mice 

compared to control mice. Remarkably, hepatic features of 

NAFLD and NASH were found in livers from VHL-KO mice 

together with increased lipid content. Accordingly, CD36 levels 

were upregulated in these mice after VHL deletion. In human 

hepatic cells, HIFs were stabilized under hypoxic conditions. 

Interestingly, hypoxia itself enhanced fatty acid uptake monitored 

by Nile Red staining due to the increase of CD36 translocation 

to the plasma membrane. This event was parallel to 

an increase of inflammatory markers. Noteworthy, palmitic-induced 

lipotoxicity was more pronounced under hypoxic conditions. 

CONCLUSIONS: Hypoxia accelerates fatty acid uptake, 

largely due to CD36 translocation to the plasma membrane of 

hepatocytes, leading to increased fat accumulation and inflammation 

in mice and in cultured human hepatocyte-derived cells. 

These results suggest that hypoxia could have a key pathogenic 

role in the progression of NAFLD to NASH. 

Disclosures: 

The following authors have nothing to disclose: Agueda Gonzalez-Rodriguez, 

Gloria Mateo, Ines Soro-Arnaiz, Mar Torres-Capelli, Ainara Elorza, Julian Aragones, 

Carmelo García-Monzón 

931 

Predicted prevalence of NAFLD and NASH in a large 

population using non-invasive multiparametric MRI 

Catherine Kelly, Rajarshi Banerjee; Perspectum Diagnostics Ltd, 

Oxford, United Kingdom 

Aim: To determine the suitability of multiparametric MRI of 

the liver for the assessment of NAFLD in large populations. 

Methods: 1000 people, aged 40-69, were recruited from the 

electoral register for multiparametric MRI, according to the 

LiverMultiScan protocol. Liver disease status was unknown. 

Estimates of liver fat fraction (PDFF %) and fibroinflammatory 

disease (LIF score) were calculated using LiverMultiScan 

software. Population statistics were compared to those in a 

previously-reported cohort (Banerjee et al. 2014) with biopsy-proven 

NASH (NAS ≥ 5). Results: In the unselected population 

sample, 19.4% had steatosis (>5% PDFF), in agreement 

with UK population estimates (Preiss & Sattar, 2008). In the reference 

NASH cohort, 100% had steatosis (mean PDFF 29.6%). 

The mean LIF score in the NASH cohort was 2.85 (s.d. 0.75). 

A LIF score > 2 is associated with a higher likelihood of liver-related 

clinical events (Pavlides, et al., 2014). In the unselected 

group, LIF ranged from 0 to 3.2 with a mean of 0.89, (s.d. 

0.3). In a plot of LIF vs PDFF, the NASH cohort can be used 

to define the upper right quadrant (PDFF > 5% and LIF > 1.1). 

Approximately 4% of the unselected cohort occupy the NASH 

quadrant, in agreement with previous estimates of NASH prevalence 

(Vernon et al. 2011). 9 of these individuals had LIF > 2. 

Conclusion: Estimates of steatosis and NASH in an unselected 

population agree with previous prevalence studies, suggesting 

that multiparametric MRI is a promising technique for the 

assessment of NAFLD. These findings encourage the use of this 

methodology in other population studies, including screening.


LIF vs. PDFF in unselected (dots) and confirmed NASH (squares) 

populations. 

932 

Akkermansia Muciniphila Is Decreased In Patients With 

Non-Alcoholic Fatty Liver Disease 

Tarkan Karakan 1 , Ceren Gundogdu 2 , Meltem Yalinaycirak 2 , Mehmet 

Ibis 1 ; 1 Gastroenterology, Gazi University, Ankara, Turkey; 

2 Medical Microbiology, Gazi University, Ankara, Turkey 

Objective The bacterial overgrowth in the intestine, disruption 

of the balance in the gut microbiota (dysbiosis) may have an 

effect on NAFLD pathogenesis. Previous reports suggest a beneficial 

role of Akkermansia muciniphila in obesity and metabolic 

syndrome. The purpose of the present study is to compare 

the gut microbiota of the patients with NAFLD and the healthy 

controls by quantitative Real Time PCR (qPCR) analysis. In 

order to understood the potential role of gut dysbiosis and subsequent 

translocation of bacterial products, serum endotoxin 

levels were also been analyzed. Methods The stool and serum 

samples from 52 NAFLD patients and 38 healthy controls have 

been collected. qPCR analysis of Akkermansia muciniphila, 

Faecalibacterium prausnitzii, Lactobacillus spp., Bifidobacterium 

spp., Bacteroides fragilis group was performed. Serum 

endotoxin levels were also determined by Chromogenic LAL 

Assay. Results Akkermansia muciniphila and Bacteroides fragilis 

group were significantly lower in patients with NAFLD as 

compared with the healthy control (p


bined with low doses of metformin and sildenafil in the treatment 

of NAFLD and NASH. 

Disclosures: 

Michael Zemel - Board Membership: NuSirt Biopharma; Management Position: 

NuSirt Biopharma; Patent Held/Filed: NuSirt Biopharma; Stock Shareholder: 

Nusirt Biopharma 

Antje Bruckbauer - Employment: NuSirt Biopharma; Patent Held/Filed: NuSirt 

Biopharma; Stock Shareholder: NuSirt Biopharma 

The following authors have nothing to disclose: Bingzhong Xue, Hang Shi 

934 

Role of fibroblast growth factor 15 in the development 

of high fat diet induced NASH 

Justin D. Schumacher 1 , Bo Kong 1 , Yang Pan 2 , Le Zhan 1 , Runbin 

Sun 2 , Jiye Aa 2 , Jason R. Richardson 1 , Debra Laskin 1 , Grace L. 

Guo 1 ; 1 Toxicology, Rutgers University, Smithtown, NY; 2 Key Laboratory 

of Drug Metabolism and Pharmacokinetics, Chinese Pharmaceutical 

University, Nanjing, China 

With the rise in obesity in Western civilizations, the prevalence 

of non-alcoholic steatohepatitis (NASH) is increasing with 

an estimated 5-10% of the population affected. The primary 

features of NASH include steatosis, inflammation, and fibrosis 

often accompanied with metabolic syndrome. Fibroblast 

growth factor 15 (Fgf15), an endocrine factor mainly produced 

in the distal part of small intestine, emerges to be a critical 

factor in regulating bile acid homeostasis, energy metabolism, 

and liver regeneration. In order to investigate the effects of 

Fgf15 on the development of each the listed features of NASH, 

Fgf15-/- mice were bred into a 75% A129 and 25% C57BL/6 

background. Four-week old Fgf15-/- and wild-type (WT) control 

mice were fed either a high fat diet (HFD) or control diet for 

six months. Body weight changes and food consumption were 

recorded regularly and a glucose tolerance test was administered 

during the fifth month. At the end of feeding, liver, intestine, 

and blood samples were collected for gene expression 

analysis, determination of serum and tissue lipid composition, 

identification of biomarker concentrations, and histology. The 

results showed that HFD-fed Fgf15-/- mice had metabolic syndrome 

presenting with increased body weight, decreased insulin 

sensitivity and increased basal total serum cholesterol levels. 

Knockout animals had altered expression of lipid metabolic 

enzymes with basal Mtp down-regulated and Acss2 up-regulated 

while on a HFD compared to controls. Fgf15 deficiency 

had no observed effects on steatosis as WT and Fgf15-/- mice 

were found to have comparable hepatic levels of triglycerides 

and total cholesterol. However, the Fgf15-/- mice were protected 

from the development of hepatic fibrosis revealed by 

histology analysis and expression of the genes involved in 

fibrosis. No changes were observed in hepatic expression of 

inflammatory genes such as Tnf-α or Icam. Lastly, Fgf15-/- mice 

fed the HFD had increased bile acid pools and up-regulated 

gene expression of ileal Ibabp and hepatic Cyp7a1, Cyp8b1, 

and Bsep compared to WT mice. In summary, during NASH 

development, Fgf15 deficiency was found to have no effects 

on liver steatosis or inflammation, however, led to decreased 

insulin tolerance, increased basal serum total cholesterol levels, 

altered expression of lipid metabolic enzymes, disrupted bile 

homeostasis, and decreased liver fibrosis. 

Disclosures: 

Grace L. Guo - Consulting: NGM 

The following authors have nothing to disclose: Justin D. Schumacher, Bo Kong, 

Yang Pan, Le Zhan, Runbin Sun, Jiye Aa, Jason R. Richardson, Debra Laskin 

935 

Fatty Acid-Stimulated Inflammasome Activation Contributes 

to Hepatic Stellate Cell Activation in High Fat/ 

Clarrie Diet-Fed Mice 

Xue-Jing Liu 1 , Na-Na Duan 1 , Ning-Ping Zhang 2,3 , Jia Ding 1,4 , 

Jian Wu 1,2 ; 1 Dept. of Pathogenic Biology/Key Lab of Molecular 

Virology, Fudan University Shanghai Medical College, Shanghai, 

China; 2 Shanghai Institute of Liver Diseases, Fudan University 

Shanghai Medical College, Shanghai, China; 3 Department 

of Gastroenterology & Hepatology, Fudan University Affiliated 

Zhongshan Hospital, Shanghai, China; 4 Department of Gastroenterology, 

Shanghai Jing’an District Central Hospital, Shanghai, 

China 

BACKGROUND: There is a significant increase in free fatty 

acids, especially saturated and oxidized fatty acids, which 

contributes to lipotoxicity, oxidant stress, and subsequent 

hepatic fibrogenesis in non-alcoholic steatohepatitis (NASH). 

However, how fatty acids lead to hepatic stellate cell activation 

is lack of experimental evidence. The present study aims to 

investigate the role of fatty-acid-mediated inflammasone activation 

in a NASH model of high fat/Clarie diet (HFC)-fed mice. 

METHODS: Mice were fed HFC diet for 13 weeks, and the 

development of NASH was assessed by liver histology. Expression 

of inflammasome molecules, NLRP1 and NLRP3 was colocalized 

in hepatocytes, Kupffer cells and hepatic stellate cells 

(HSCs) in liver section by immunohistochemistry. Palmitic acid 

(PA, an oxidized fatty acid)-stimulated activation of inflammasome 

molecules and subsequent activation in two human 

HSC lines were determined by quantitative RT-PCR. RESULTS: 

The body weight and liver weight were significantly increased 

in HFC diet-fed mice compared to controls (48.0±1.40 vs. 

30.1±0.60g, p


936 

Ancestral starvation-activated caspase-2 promotes obesity, 

the metabolic syndrome and Nonalcoholic fatty 

liver disease 

Mariana V. Machado 1,2 , Gregory A. Michelotti 1 , Guanhua Xie 1 , 

Thiago A. Pereira 1 , Mark Jewell 1 , Richard T. Premont 1 , Anna Mae 

Diehl 1 ; 1 Division of Gastroenterology, Duke University Medical 

Center, Durham, NC; 2 Gastrenterologia e Hepatologia, Hospital 

de Santa Maria, CHLN, Lisbon, Portugal 

Background: Obesity is an increasingly prevalent risk factor for 

common metabolic disorders such as the metabolic syndrome 

(MetS). The MetS promotes nonalcoholic fatty liver disease 

(NAFLD). One hypothesis for obesity-related systemic dysfunction 

is that overtaxed adipocytes release fatty acids and 

pro-inflammatory factors into the circulation to trigger insulin 

resistance, changes in pancreatic beta cells, and other manifestations 

of the MetS, including diabetes mellitus (DM) and 

NAFLD. Caspase-2 is an ancestral caspase that is activated 

by fatty acid accumulation to eliminate dispensible cells during 

starvation, thereby reducing net energy requirements. Aim: To 

evaluate the hypothesis that caspase-2 activation is a critical 

link in diet-induced MetS that leads to DM and NAFLD Methods: 

Caspase-2 deficient mice and congenic wild type mice 

were fed a Western diet (high fat diet enriched with saturated 

fatty acids and supplemented with 0.2% cholesterol + fructose 

and glucose in the drinking water) for 16 weeks. Metabolic 

and hepatic outcomes were evaluated. Results: Caspase-2 deficient 

mice fed an obesogenic Western diet were protected from 

visceral fat deposition, glucose intolerance/insulin resistance, 

pancreatic beta cell hyperplasia, dyslipidemia, and NAFLD. 

Adipose tissue in caspase-2 deficient mice was more proliferative, 

up-regulated mitochondrial uncoupling proteins, and 

was resistant to cell hypertrophy and cell death, suggesting 

that caspase 2 deficiency caused “browning” of white adipose 

tissue. Compared to congenic controls fed the Western diet, 

hepatic accumulation of nonesterified fatty acids was reduced, 

de novo lipogenesis was decreased, fatty acid b-oxidation and 

lipoprotein export were increased in caspase-2 deficient mice 

fed Western diets. Conclusion: Caspase-2 promotes lipoapoptosis 

in adipocytes and hepatocytes, resulting in the MetS, 

DM, and NAFLD. Given that genetic deficiency of caspase-2 

protects mice from the MetS and MetS-related tissue damage, 

caspase-2 is a potential therapeutic target for the treatment of 

NAFLD. 

Disclosures: 

The following authors have nothing to disclose: Mariana V. Machado, Gregory 

A. Michelotti, Guanhua Xie, Thiago A. Pereira, Mark Jewell, Richard T. Premont, 

Anna Mae Diehl 

937 

Antihypertensive therapy improves insulin resistance 

and imbalances of interleukin-6 and -10 in spontaneously 

hypertensive rats with steatohepatitis 

Masaya Kozono 1 , Hirofumi Uto 2,1 , Rie Ibusuki 3,1 , Shiho Arima 1 , 

Kohei Oda 1 , Sho Ijuin 1 , Hiroka Onishi 1 , Haruka Sakae 1 , Kaori 

Muromachi 1 , Akihiko Oshige 1 , Seiichi Mawatari 1 , Tsutomu 

Tamai 1 , Akihiro Moriuchi 4 , Hirohito Tsubouchi 5 , Akio Ido 1 ; 1 Digestive 

and Lifestyle Diseases, Department of Human and Environmental 

Sciences, Kagoshima University Graduate School of Medical 

and Dental Sciences, Kagoshima, Japan; 2 Center for Digestive 

and Liver Diseases, Miyazaki Medical Center Hospital, Miyazaki, 

Japan; 3 Department of International Island and Community Medicine, 

Kagoshima University Graduate School of Medical and 

Dental Sciences, Kagoshima, Japan; 4 Department of HGF Tissue 

Repair and Regenerative Medicine, Kagoshima University Graduate 

School of Medical and Dental Sciences, Kagoshima, Japan; 

5 Kagoshima City Hospital, Kagoshima, Japan 

Objective: Nonalcoholic steatohepatitis (NASH) is a hepatic 

manifestation of the metabolic syndrome. Features of the metabolic 

syndrome such as insulin resistance (IR) and hypertension 

are risk factors for both advanced liver disease and cardiovascular 

disease in patients with NASH. Immune factors also play 

a key role in the pathogenesis of NASH, IR, and hypertension. 

However, the molecular mechanisms of these disorders and 

the association between them have not been fully elucidated. 

In this study, we investigated the effects of severe hypertension 

induced by a high-salt diet on the pathological condition of 

spontaneously hypertensive rats (SHRs) with steatohepatitis. 

Methods: Steatohepatitis was induced by a choline-deficient, 

L-amino acid–defined (CDAA) diet. Seven-week-old male SHRs 

were randomly divided into five groups: those receiving six 

weeks of standard chow with normal salt concentration, followed 

by an additional 8 weeks of a standard chow or CDAA 

diet with normal salt concentration (control and CDAA groups, 

respectively); and those receiving six weeks of standard chow 

with high-salt (HS) concentration, followed by a CDAA diet 

containing high salt for an additional 8 weeks with or without 

the anti-hypertensive agents amlodipine (Aml) or hydralazine 

(Hyd) (CDAA+HS, CDAA+HS+Aml, and CDAA+HS+Hyd 

groups, respectively). Results: In the CDAA and CDAA+HS 

groups, blood pressure was significantly correlated with serum 

levels of alanine aminotransferase (ALT), alkaline phosphatase 

(ALP), fasting blood glucose, serum insulin, and homeostasis 

model assessment-IR (HOMA-IR). Anti-hypertensive therapy 

significantly ameliorated elevated ALP, glucose, insulin, and 

HOMA-IR. Furthermore, increased serum interleukin (IL)-6 

levels following the high-salt CDAA diet were attenuated by 

anti-hypertensive therapy. Additionally, serum IL-10 levels were 

increased by anti-hypertensive therapy, and the decreased proportion 

of CD4+CD25+ T cells and CD4+CD25+Foxp3+ T 

cells observed following a high-salt CDAA diet tended to be 

restored by amlodipine. Conclusion: These results demonstrate 

that anti-hypertensive therapy improved glucose metabolism 

and imbalances of cytokine expression in a rat model of hypertension 

with steatohepatitis, suggesting that anti-hypertensive 

therapy acting through immunological factors may be beneficial 

for patients with hypertension–associated NASH. 

Disclosures: 

The following authors have nothing to disclose: Masaya Kozono, Hirofumi Uto, 

Rie Ibusuki, Shiho Arima, Kohei Oda, Sho Ijuin, Hiroka Onishi, Haruka Sakae, 

Kaori Muromachi, Akihiko Oshige, Seiichi Mawatari, Tsutomu Tamai, Akihiro 

Moriuchi, Hirohito Tsubouchi, Akio Ido


938 

Alterations in NK cell phenotype with disease progression 

in a murine model of non-alcoholic fatty liver disease 

(NAFLD) 

Rachel McMahan, Cara Porsche, Lucy Golden-Mason, Hugo R. 

Rosen; Gastroenterology, University of Colorado, Aurora, CO 

Background: Nonalcoholic fatty liver disease (NAFLD) is associated 

with a spectrum ranging from benign accumulation of triglycerides 

within the cytoplasm of liver hepatocytes (steatosis) 

to a more severe disease, nonalcoholic steatohepatitis (NASH). 

While the function of NKT cells in NAFLD progression has been 

extensively investigated, there are limited data on the role of 

NK cells in this disease. To further clarify the role of these cells 

in NAFLD we analyzed NK cell phenotype and function over 

time in a murine model of NAFLD. Methods: To investigate the 

role of NK cells during NAFLD disease progression we established 

a murine model of NAFLD/NASH demonstrating the full 

disease spectrum. Mice were fed high fat, high cholesterol, high 

sucrose diet containing trans-fats from 8 weeks to 20 weeks. 

NAFLD severity at 8, 12 and 20 weeks was evaluated by 

liver histology and intrahepatic gene expression. NK cells were 

isolated from livers at the indicated time points and stained 

for cell surface markers and degranulation. Results: We have 

established a murine progressive NAFLD model mimicking the 

disease spectrum observed in NASH/NAFLD. After 8 weeks of 

feeding the mice had increased hepatic steatosis without significant 

inflammation while at the 12 week time point inflammatory 

foci were found along with increased macrovesicular steatosis. 

By 20 weeks, the mice had also developed significant fibrosis 

along with inflammation and steatosis mimicking NASH. As 

disease progressed from steatosis to steatohepatitis intrahepatic 

NK cells significantly increased expression of activation 

markers (CD69, p


C57BL6 wild-type or RIP3-deficient (RIP3 -/- ) mice were fed highfat 

choline-deficient (HFCD) or methionine and choline-deficient 

(MCD) diets, with subsequent histological and biochemical 

analysis of hepatic damage. In primary murine hepatocytes, 

necroptosis and oxidative stress were also assessed after necrostatin-1 

treatment or RIP3 silencing. We show that in chronic 

liver disease patients, RIP3 levels were significantly increased 

and correlated with steatohepatitis histological severity (at least 

p < 0.05). Circulating markers of necrosis and TNF-α, as well 

as liver MLKL phosphorylation were increased in NAFLD (at 

least p < 0.05). Likewise, RIP3 and MLKL protein levels and 

TNF-α expression were increased in the liver of HFCD and 

MCD diet-fed mice (at least p < 0.05). Moreover, RIP3 and 

MLKL sequestration in the insoluble protein fraction of NASH 

mice liver lysates, strongly suggesting necroptosis activation, 

represented an early event during stetatohepatitis progression 

(at least p < 0.05). Functional studies in primary murine hepatocytes 

established the association between TNF-α-induced RIP3 

expression, activation of necroptosis and oxidative stress (p < 

0.05). Strikingly, RIP3 deficiency attenuated MCD diet-induced 

liver injury, steatosis, inflammation, fibrosis and oxidative stress 

(at least p < 0.05). In conclusion, necroptosis is increased in 

the liver of NAFLD patients and in experimental models of 

NASH. Further, TNF-α triggers RIP3-dependent oxidative stress 

during hepatocyte necroptosis. As such, targeting necroptosis 

appears to arrest or at least impair NAFLD progression. 

Disclosures: 



The following authors have nothing to disclose: Marta B. Afonso, Pedro M. 

Rodrigues, Tânia Carvalho, Marta Caridade, Paula Borralho Nunes, Rui E. Castro, 

Cecília M. Rodrigues 

941 

Over-expression of HNF4α attenuated lipotoxicity in 

non-alcoholic fatty liver disease model 

Jai Sun Lee 1 , Dae Won Jun 2 , Seung Min Lee 3 , Yong Kyun Cho 4 , 

Eun Chul Jang 5 , Sang Bong Ahn 6 ; 1 Department translational medicine, 

Hanyang University Graduate School of Biomedical Science 

and Engineering, Seoul, Korea (the Republic of); 2 Department 

of Internal Medicine, Hanyang University College of Medicine, 

Seoul, Korea (the Republic of); 3 Department of Food and Nutrition, 

Sungshin Women’s University, Seoul, Korea (the Republic 

of); 4 Department of Internal Medicine, Kangbuk Samsung Hospital, 

Sungkyunkwan University, School of Medicine, Seoul, Korea 

(the Republic of); 5 Department of Occupational and Environmental 

Medicine, Soonchunhyang University Cheonan Hospital, Cheonan, 

Korea (the Republic of); 6 Department of Internal Medicine, Eulji 

University School of Medicine, Seoul, Korea (the Republic of) 

Background: Hepatocyte nuclear factor 4α (HNF4α) is known 

as a master regulator of liver-specific gene expression. The 

effects of HNF4α on non-alcohol fatty liver disease (NAFLD) 

at transcriptional level are largely unknown. In this study, we 

evaluated the role of HNF4α in NAFLD model. Methods: High 

fat (HF) diet was fed to rats for 20 weeks to induce NAFLD. To 

make vitro NAFLD model, palmatic acid (PA) treated in HepG2 

cells for 24 hours. Later, liver-specific gene expressions were 

evaluated. Using microporator, a transcription factor, HNF4α 

was over-expressed in HepG2 cells. After transfection, MTT 

assay, Tunnel assay, FACS, Nile-red staining, and qPCR gene 

evaluation were performed. Results: Hepatic gene expression 

of Foxa2 and GATA4 were up regulated in HF diet fed rats 

compared to control group. But HNF4α mRNA expression was 

decreased in diet induced fatty disease model. In HepG2 cells, 

palmatic acid treatment also decreased HNF4α mRNA expression; 

however, interestingly, the oleic acid treatment did not 

affect the HNF4α mRNA expression. HepG2 cells over-expressing 

HNF4α through transfection showed a protective effect 

against palmatic acid-induced apoptosis. HNF4α over-expression 

was also associated with increased fatty acid oxidation 

and VLDL secretion. The HepG2 cells over-expressing HNF4α 

also exhibited increased the mRNA expression of enzymes 

related to VLDL secretion (MTTP and ApoB) and fatty acid oxidation 

(ACOX and CYP4A1). Moreover HNF4α over-expression 

was also involved in decreased fatty acid uptake. The 

HepG2 cells over-expressing HNF4α also exhibited decreased 

the mRNA expression of fatty acid uptake mediator (CD36 

and FATP2). Conclusions: HNF4α attenuated lipotoxicity by 

increased VLDL secretion and fatty acid oxidation in NASH 

model. 

Disclosures: 

The following authors have nothing to disclose: Jai Sun Lee, Dae Won Jun, Seung 

Min Lee, Yong Kyun Cho, Eun Chul Jang, Sang Bong Ahn 

942 

Thioesterase Superfamily Member 2 (Them2) Reduces 

Membrane Fluidity and Promotes Endoplasmic Reticulum 

(ER) Calcium Loss in Response to Saturated Free 

Fatty Acids: Pathogenic Role in Non-Alcoholic Fatty Liver 

Disease (NAFLD) 

Baran A. Ersoy, Yingxia Li, David E. Cohen; Medicine, Brigham 

and Women’s Hospital and Harvard Medical School, Boston, MA 

Background: NAFLD is associated with maladaptive increases 

in hepatic glucose production in the setting of excessive nutrition 

that are due in part to insulin resistance. Saturated free 

fatty acids (FFA) contribute by reducing ER membrane fluidity. 

This promotes insulin resistance, which is attributable in part to 

depletion of ER calcium and ER stress. In addition, increased 

cytosolic calcium levels promote hepatic glucose production 

by activating calcium/calmodulin-dependent protein kinase 2 

(CaMKII) and amp-activated protein kinase (AMPK). Genetic 

ablation of Them2, an acyl-CoA thioesterase that preferentially 

converts saturated fatty acyl-CoAs to FFA, protects against 

depletion of ER calcium and ER stress in cell culture systems and 

reduces hepatic glucose production in high fat fed mice. Aim: 

This study was designed to determine the molecular mechanism 

by which Them2 expression regulates ER calcium homeostasis 

and hepatic glucose production. Methods: HEK 293E cells 

were exposed for 6 h to 0.5 mM palmitic acid after treatment 

with Them2 or scrambled (control) siRNA. ER calcium release 

and reuptake were measured using Fluo-4 following treatment 

of cells with the re-uptake inhibitor thapsigargin (2 mM). Total 

ER calcium stores were measured following treatment of cells 

with 5 mM ionomycin, an ionophore that creates calcium-permeable 

membrane pores. Membrane fluidity was measured by 

polarization anisotropy of diphenylhexatriene, as well as by 

formation of pyrenedecanoic acid eximers using fluorescence 

spectrometry. Activation of calcium-sensitive phosphoproteins 

that promote hepatic glucose production was assessed by 

immunoblot analysis. Results: In the absence of palmitic acid 

treatment, Them2 knockdown reduced thapsigargin-mediated 

loss of ER calcium by 40%. This was not attributable to reduced 

ER calcium stores because knockdown of Them2 expression 

did not alter ionomycin-mediated calcium release. Palmitic acid 

treatment reduced ER membrane fluidity by 58%, reduced ER 

calcium uptake by 11% and increased cytosolic calcium by 

2-fold. These effects were abrogated by knockdown of Them2. 

Moreover, reductions in cytosolic calcium following Them2 

knockdown correlated with decreases in activation of both 

CaMKII and AMPK. Conclusions: Them2 mediates the effects of 

saturated FFA on ER membrane fluidity and calcium depletion,


which in turn lead to activation of CaMKII and AMPK. These 

findings identify a likely mechanism by which Them2 promotes 

hepatic glucose production in response to high fat feeding and 

provide a rationale for targeting Them2 in the management of 

NAFLD. 

Disclosures: 



The following authors have nothing to disclose: Baran A. Ersoy, Yingxia Li 

943 

Genetic analysis of NAFLD within a Caribbean Hispanic 

Population 

Devorah Edelman 2 , Bernice Morrow 2 , Maria Delio 2 , Mortadha 

Abd 4 , Adam Auton 2 , Tao Wang 3 , Allan W. Wolkoff 1,4 , Harmit S. 

Kalia 1,4 ; 1 Medicine, Albert Einstein College of Medicine, Bronx, 

NY; 2 Genetics, Albert Einstein College of Medicine, Bronx, NY; 

3 Epidemiology, Albert Einstein College of Medicine, Bronx, NY; 

4 Gastroenterology & Liver Diseases, Montefiore Medical Center, 

Bronx, NY 

We performed the first genetic investigation of Non-alcoholic 

fatty liver disease (NAFLD) in Hispanic patients of majority 

Caribbean descent. A total of 316 individuals including 40 

subjects with biopsy-proven NAFLD, 24 ethnically matched 

non-NAFLD controls, and a 252 ethnically-mixed random sampling 

of Bronx County, New York were analyzed. Genotype 

analysis was performed to determine allelic frequencies of 234 

known single nucleotide polymorphisms (SNPs) associated with 

NAFLD risk based on previous genome-wide association study 

(GWAS) and candidate-gene studies. Additionally, the entire 

coding region of PNPLA3, a gene showing the strongest association 

to NAFLD was subjected to Sanger sequencing to identify 

all exonic variants in our NAFLD samples and controls. The 

highly NAFLD-correlated rs738409 polymorphism in PNPLA3 

occurred more frequently in the case population than controls 

(OR 2.95, p=0.003), the general population (OR 1.97, 

p=0.0001), and would be expected in a random Puerto Rican 

population (OR 1.56, p= 0.05) based on data from the 1000 

Genomes Project. Results suggest that both rare and common 

DNA variations in PNPLA3 and SAMM50 may be correlated 

with NAFLD in this small population study, while common DNA 

variations in CHUK and ERLIN1, may have a protective interaction. 

Common SNPs in ENPP1 and ABCC2 have suggestive 

association with fatty liver, but with less compelling significance. 

In conclusion, Hispanic patients of Caribbean ancestry 

may have different interactions with NAFLD genetic modifiers; 

therefore, further investigation with a larger sample size, into 

this Caribbean Hispanic population is warranted. 

Genotyping Results 

Disclosures: 


The following authors have nothing to disclose: Devorah Edelman, Bernice Morrow, 

Maria Delio, Mortadha Abd, Adam Auton, Tao Wang, Harmit S. Kalia 

944 

Inhibitor Of DNA Binding 2 Gene Induced By High Levels 

Of Insulin Contributes To Adipogenesis And Fatty 

Liver In A Model Of Type 2 Diabetes Mellitus 

Tomoyuki Nemoto, Hidetaka Matsuda, Katsushi Hiramatsu, Yoshihiko 

Ozaki, Tatsushi Naito, Kazuto Takahashi, Kazuya Ofuji, 

Masahiro Ohtani, Hiroyuki Suto, Yasunari Nakamoto; University 

of Fukui, Fukui, Japan 

BACKGROUND: Hyperinsulinemia is well known to be associated 

with type 2 diabetic patients. It remains unclear how 

hyperinsulinemia contributes to progression of obesity, pathogenesis 

of fatty liver and development of non-alcoholic steatohepatitis 

(NASH). In the previous studies, inhibitor of DNA 

binding 2 (Id2) gene, a dominant-negative transcriptional 

repressor, has been shown to function as a promoting factor 

for obesity. Here, we examined, in vitro and in vivo, whether 

hyperinsulinemia causes promotion of obesity and pathogenesis 

of fatty liver mediated by Id2 using type 2 diabetes 

mellitus models. METHODS: Mouse fibroblast NIH-3T3 cells 

and human hepatoma HepG2 cells were treated with various 

concentrations of insulin, and Id2 mRNA and protein expression 

were analyzed by Northern and Western blots. Mouse 

preadipocyte 3T3-L1 cells were differentiated to adipocytes in 

differentiation cocktail (insulin, 3-isobutyl-1-methylxanthine and 

dexamethasone). KK-A y mice were used as a type 2 diabetes 

mellitus model. Blood glucose was measured using a glucose 

monitor, and serum insulin was measured by ELISA. Histological 

examination was performed on liver specimens. KK-A y mice 

were crossed with Id2 KO mice, and KK-A y -Id2 KO mice were 

obtained. Mice had body weight recorded weekly. RESULTS: 

Id2 expression at mRNA and protein levels in NIH-3T3 cells 

and HepG2 cells was induced by insulin in a dose-dependent 

manner. Id2 induction by insulin was partially inhibited by PI3 

kinase inhibitor. 3T3-L1 cells expressed high amount of Id2 

protein by addition of differentiation cocktail containing insulin 

and differentiated to adipocytes. In contrast, the cells did 

not differentiate by the cocktail lacking insulin. In KK-A y mice, 

blood glucose and serum insulin (3.0 ng/ml) was significantly 

higher compared to non-diabetic 129/Sv mice (0.58 ng/ml) 

(P


945 

Human amniotic epithelial cells reduce liver fibrosis 

and the incidence of hepatocellular carcinoma in a ‘fast 

food’ NAFLD model 

Alexander Hodge 1 , Nathan T. Kuk 1 , Ying S. Sun 1 , Kristy Nguyen 1 , 

Rebecca Lim 2 , William Sievert 1 ; 1 Monash Medical Centre,Monash 

University, Clayton, VIC, Australia; 2 Hudson Institute of Medical 

Research, Melbourne, VIC, Australia 

Non-alcoholic fatty liver disease (NAFLD) is associated with 

obesity and can lead to liver cirrhosis and hepatocellular carcinoma 

(HCC). Currently there are no effective treatments except 

lifestyle modification. Sourced from human placentas, human 

amniotic epithelial stem cells (hAEC) and hAEC-conditioned 

media (hAEC-CM) have anti-inflammatory and anti-fibrotic 

properties. We examined the efficacy of these cells in a NAFLD 

model. Methods: C57BL/6J mice received a ‘fast food’ diet 

(FFD) (21% fat, 2% cholesterol with 42g/L fructose drinking 

water). At week 34, mice were given one intraperitoneal (IP) 

injection of 2x10 6 hAEC (group 1); two injections of 2x10 6 

hAEC four weeks apart (group 2) or thrice weekly IP 400mL 

hAEC-CM (group 3). Controls were untreated. Mice were 

culled at week 42. Liver fibrosis area was determined by sirius 

red staining. Hepatic stellate cell activation was determined by 

qPCR for alpha-smooth muscle actin (α-SMA). Hepatic inflammation 

was measured by serum ALT and staining for liver macrophages 

(F4/80 + ). Livers were evaluated macroscopically 

for the presence of HCC and confirmed histologically. Liver 

progenitor cell (LPC) numbers were assessed by pan-CK staining. 

Computer assisted morphometry was used to quantify the 

percentage of positively stained liver (expressed as % area 

of liver tissue). Results: Controls showed typical pericellular 

hepatic fibrosis and steatosis. Liver fibrosis area was reduced 

by 40% in all treatment groups (p


947 

Adipose tissue-derived stromal cells preventatively suppressed 

pathological progression of murine non-alcoholic 

steatohepatitis model. 

Masatoshi Yamato 1,2 , Yoshio Sakai 1 , Hatsune Mochida 2 , Akihiro 

Seki 2 , Kosuke Ishida 2 , Masao Honda 2,1 , Shuichi Kaneko 2,1 ; 1 Gastoroenterology, 

Kanazawa University, Kanazawa, Japan; 2 Disease 

Control and Homeostasis, Kanazawa University, Kanazawa, 

Japan 

Non-alcoholic steatohepatitis (NASH) is characterized by steatosis 

in hepatocytes accompanied with persisting inflammation, 

developing fibrosis to ultimate cirrhosis. The pathogenesis 

of NASH is not fully elucidated, and the fundamental treatment 

of NASH is yet to be established. We previously reported that 

ADSCs are therapeutically beneficial as being functionally and 

histologically repairable of the end-stage cirrhotic mice caused 

by NASH. In this study, we analyzed pathological progression 

of NASH mice model and succeedingly assessed how 

ADSC administration affected on early stage NASH disease 

progression.[Materials and Methods] C57Bl/6J mice (female, 

10 weeks old) were initiated to be fed with high fat atherogenic 

(HF-AT) diet to develop steatohepatitis. On week 1, 2, 

3, 4, 8, and 12, we obtained liver tissue samples. ADSCs 

were isolated from adipose tissue of C57BL/6J mice by collagenase 

I digestion, cultured and expanded. On week 4 and 8, 

ADSCs (1×10^5 ) were injected twice every 4 weeks into the 

splenic subcapsule, and on week 12, the liver tissue samples 

were obtained. These liver tissues were analyzed for gene 

expressions by real-time detection PCR (RTD-PCR) and immunohistochemistry.[Results]Hepatocyte 

steatosis was observed 

from 1 week after initiation of HF-AT feeding, not accompanied 

with infiltration of inflammatory cells until 4 week. On week 4, 

Gr-1+ inflammatory cells were initially found to be infiltrated 

into the portal area and had continuously increased with time 

until week 12. TNF-α gene expression of the liver tissue was 

also increased consistently with increase of hepatic infiltration 

of Gr-1+ cells. Obvious fibrosis had been observed since 

week 8. After 8 week, liver fibrosis was extended, and gene 

expressions of albumin and type IVa collagen was decreased 

and increased, respectively. When ADSCs were injected into 

NASH mice, the fibrosis area was less extended on week 12 

than that of PBS injection mice group. RTD-PCR analysis of 

hepatic tissue showed the substantial maintained albumin gene 

expression (p=0.04) and the significant decrease level of Collagena 

(p=0.04) and TNFα(p=0.04) gene expressions, which 

suggested that ADSC improved protein production, suppressing 

fibrosis progression, and hepatic inflammation.[Conclusion] 

ADSC treatment prevented pathological progression of liver 

fibrosis with persisting hepatic inflammation, and prevented the 

decrease of albumin production of the liver during early phase 

of NASH development. ADSC treatment is considered to be 

preventively and therapeutically efficacious in part to NASH. 

Disclosures: 






The following authors have nothing to disclose: Masatoshi Yamato, Yoshio Sakai, 

Hatsune Mochida, Akihiro Seki, Kosuke Ishida, Masao Honda 

948 

High and Low Capacity Runner (HCR and LCR) rat lines 

do not display significant differences in NASH phenotype 

when fed a fast food diet 

Howard C. Masuoka 1,3 , Oscar Cummings 2 , Naga P. Chalasani 1,3 , 

Lawrence Lumeng 1 ; 1 Medicine, Division of Digestive and Liver DIsorders, 

Indiana University, Indianapolis, IN; 2 Pathology and Laboratory 

Medicine, Indiana University, Indianapolis, IN; 3 Cellular 

and Integrative Physiology, Indiana University, Indianapolis, IN 

Exercise and dietary changes are cornerstones of treatment 

of non-alcoholic fatty liver disease (NAFLD) including non-alcoholic 

steatohepatitis (NASH). Sedentary lifestyle is felt to 

predispose individuals to NAFLD/NASH, and improved cardiorespiratory 

fitness is associated with lower hepatic steatosis. 

High and Low Capacity Runner (HCR and LCR) are established, 

extensively phenotyped rat lines generated by selective breeding 

for contrasting extremes in maximum aerobic-exercise 

capacity (MAC). Relative to the HCR, LCR rats exhibit greater 

visceral adiposity, insulin resistance, and levels of circulating 

pro-inflammatory hormones but lower home cage activity, and 

non-exercise activity thermogenesis. We hypothesized that LCR 

rats will display increased features of NASH relatively to HCR 

rats when fed a “fast food” (FF) diet high in saturated fat, 

cholesterol and fructose. In mice this diet better recapitulates 

features of human NASH compared to a high fat diet. We had 

custom formulated a FF diet containing a caloric density of 4.61 

kcal/gram with a caloric composition of 44.9% fat, 15.1% 

protein, and 39.9% carbohydrates. The diet was nutritionally 

complete, and contained 30% fructose, 14% lard, 8% coconut 

oil, and 2% cholesterol. For 16 weeks 10 LCR and 10 HCR 

male rats were fed the FF diet ad libitum. Fasting weights were 

obtained weekly. Fasting serum ALT, glucose and insulin were 

measured every 4 weeks. MRI images were obtained monthly 

to determine adipose, lean, and fluid mass. As expected, LCR 

rats gained significantly more weight than HCR rats (P


949 

High calories intake and particulate matter exposition 

promote the progression from steatosis to steatohepatitis 

and create a permissive enviroment for hepatocellular 

carcinoma development 

Mirko Tarocchi, Giada Marroncini, Simone Polvani, Sara Tempesti, 

Tommaso Mello, Francesca Zanieri, Elisabetta Ceni, Andrea 

Galli; Department of Experimental and Clinical Biomedical Sciences, 

University of Florence, Florence, Italy 

Introduction: Nonalcoholic fatty liver disease (NAFLD) is 

becoming the most common chronic liver disease, and the prevalence 

is rapidly increasing in developed countries. Nonalcoholic 

steatohepatitis (NASH), the severe form of NAFLD, can 

also progress to liver cirrhosis and hepatocellular carcinoma. 

Recent evidences suggest that environmental factors can trigger 

hepatic inflammation and progression of steatosis to NASH. 

Aim: We evaluate if a western style diet in association with 

chronic urban particulate matter exposition can modifies the 

pathogenesis and progression of NASH, and support hepatocellular 

carcinoma development. Materials and methods: The 

experimental model was created to reproduce urban lifestyle: 

C57Bl/6 mice were fed with a western style diet (HFD), and 

treated with particular matter (PM) collected from the urban 

area of Florence (Italy). After 4 and 8 weeks were performed 

the morphologic analysis of liver tissues, the evaluation of 

inflammatory cell infiltrate and the collagen deposition; we 

evaluate also the effects of PM on cytokine production and oxidative 

stress related cellular damage. After 8 weeks part of the 

animals was also injected with murine hepatoma cells (2x10 6 

Hepa1-6) and sacrificed after additional 2 weeks. Results: Both 

the HFD groups developed fat accumulation in the liver, and 

at 8 weeks, the NASH score was significantly increased in 

HFD-PM group (p


951 

Fermented soymilk prevent free fatty acid-induced lipogenesis 

and production of reactive oxygen species in 

hepatocellular steatosis model 

Sang Bong Ahn 1 , Byoung Kwan Son 1 , Dae Won Jun 2 , Yong Kyun 

Cho 3 ; 1 Gastroenterology, Eulji University, School of Medicine, 

Seoul, Korea (the Republic of); 2 Internal Medicine, Hanyang 

University, College of Medicine, Seoul, Korea (the Republic of); 

3 Internal Medicine, Kangbuk Samsung Hospital, Sungkyunkwan 

University, School of Medicine, Seoul, Korea (the Republic of) 

INTRODUCTION: Ingredients of soy and fermented products 

have been widely utilized as food supplement for health-enhancing 

properties, such as reducing the risk of osteoporosis, 

protection of cardiovascular diseases, and prevention of prostate 

and breast cancer. This study was carried out to examine 

the effects of fermented soymilk (FSM) on the free fatty acid-induced 

lipogenesis in an in vitro model of hepatocellular steatosis 

model. MATERIALS AND METHODS: HepG2 cells were 

incubated with 0.2 mM of palmitic acid (PA) for 24 h to induce 

lipogenesis and to accumulate the intracellular lipid accumulation, 

which was observed by oil red O and Nile red staining. 

The PA treated cells were co-incubated with 0.04~1.0% of 

lyophilized FSM, 0.05 mM of genistein, and 50 nM of estrogen, 

respectively. Western blot analysis of sterol regulatory element-binding 

protein-1 (SREBP-1) and nuclear factor erythroid 

2-related factor-2 (NRF-2) were performed to examine the lipogenesis 

related extracellular signal-regulated kinase (ERK) pathway. 

Cellular reactive oxygen species (ROS) was measured 

by the DCFDA assay kit. RESULTS: Lipid accumulations in the 

PA and FSM co-incubated cells were significantly decreased 

by 0.5% and 1.0% of FSM without cytotoxicity. Treatments of 

PA and combining with genistein and estrogen significantly 

increased the expressions of SREBP-1. However, FSM co-incubation 

significantly attenuated the expression of SREBP-1 

in the PA treated cells. In addition, expression of NRF-2 and 

phosphorylation of ERK were significantly increased in the PA 

and FSM co-incubated cells. PA induced ROS production was 

significantly reduced by 1.0% of FSM. Meanwhile, genistein 

or estrogen alone did not lead to significant differences in ROS 

production. CONCLUSION: Our results show that bioactive 

components, except genistein and phytoestrogen, in fermented 

soymilk protect hepatocytes against lipid accumulation and 

ROS production induced by free fatty acid. These effects may 

be mediated by inhibition of SREBP-1 and activation of NFR-2 

via ERK pathway in hepatocytes. 

Disclosures: 

The following authors have nothing to disclose: Sang Bong Ahn, Byoung Kwan 

Son, Dae Won Jun, Yong Kyun Cho 

952 

Increased 53-binding protein 1 nuclear focus expression 

in patients with non-alcoholic steatohepatitis 

Koji Okamoto 2 , Yuko Akazawa 4 , Hisamitsu Miyaaki 4 , Satoshi 

Miuma 4 , Naota Taura 4 , Hisayoshi Kondo 3 , Masahiro Nakashima 1 , 

Kazuhiko Nakao 4 ; 1 Department of tumor and diagnostic pathology, 

Atomic bomb disease institute, Nagasaki, Japan; 2 Nagasaki 

University School of Medicine, Nagasaki, Japan; 3 Biostatistics Section, 

Division of Scientific Data Registry, Department of Radioisotope 

Medicine, Atomic Bomb Disease Institute, Nagasaki, Japan; 

4 Gastroenterology and Hepatology, Nagasaki University Hospital, 

Nagasaki, Japan 

Background : Pathogenesis of Non-alcoholic fatty liver disease 

(NAFLD) and the mechanisms of HCC development remain 

largely unclear. Elevated DNA damage response can result 

in genomic instability, which may lead to transformation to 

cancer. The p53-binding protein 1 (53-BP1) localizes at the 

sites of DNA double-strand breaks by irradiation and rapidly 

forms nuclear foci. We have reported that increased number 

and size of 53-BP1 nuclear foci expression represents the level 

of sporadic genomic instability during cervical carcinogenesis 

(Matsuda K, Histopathology. 2011,441-51). However, the 

presence of genomic instability in the NAFLD liver, especially 

the expression pattern of 53-BP1, is largely uninvestigated. 

Our aim was to assess the double-strand breaks based on 

53-BP1 expression in hepatocytes of NAFLD liver and free fatty 

acid-treated hepatocytes. Methods: Isolated rat primary hepatocytes 

were exposed to saturated free fatty acid palmitate (200 

μM) for 16 hour. In addition, 23 human liver biopsy samples, 

including 5 from normal livers, 5 from simple steatotic livers, 

and 13 from livers with NASH, were examined. Cells and 

biopsy samples were studied by using co-immunofluorescence 

with the anti-53-BP1 and hepatocyte marker (hepatocyte). The 

number of 53-BP1 nuclear foci in hepatocytes and its association 

with clinical features including levels of aspartate aminotransferase 

(AST), alanine aminotransferase (ALT), platelet (PLT), 

serum collagen type 4, and serum hyaluronic acid were then 

examined. Results: The palmitate treatment in the rat primary 

hepatocytes resulted in a significant increase in the number of 

53-BP1 nuclear foci (6.01% vs 24.5%, control vs palmitate, 

p< 0.05). Furthermore, in the human liver biopsy tissue samples, 

the number of 53-BP1 nuclear foci in the hepatocytes was 

increased by approximately 6 folds in the NASH livers compared 

to the simple steatotic livers, and approximately 10 folds 

compared to normal controls (p < 0.01). 53-BP1 nuclear foci 

were evenly distributed throughout zone 1 (portal area), zone 

2, and zone 3 (hepatic vein area) in the NASH liver. The rate 

of hepatocytes with more than two 53-BP1 foci/ nucleus was 

positively associated with age (p


bution. Mat1a encodes the enzyme mainly expressed in normal 

liver. Mat1a ablation in mice results in the spontaneous 

development of non-alcoholic steatohepatitis (NASH). We 

observed that SAMe depletion in Mat1a KO mice had three 

main effects on hepatic lipid metabolism: 1) impaired TG (triglyceride) 

export via VLDL; 2) impaired mitochondrial FA (fatty 

acid) oxidation (as evidenced by membrane depolarization, 

downregulation of Phb1 (prohibitin 1, a mitochondrial chaperone 

protein) and Mcj/Dnajc15 (endogenous mitochondrial 

repressor of respiratory chain), and accumulation of long-chain 

acylcarnitines); and 3) increased FA uptake. The convergence 

of these three factors induced TG accumulation in LD (lipid 

droplets). LD expansion confronts hepatocytes with a high 

demand of PC (phosphatidylcholine) molecules to cover the LD 

surface since other phospholipids, such as PE (phosphatidylethanolamine), 

cannot stabilize LD and prevent coalescence. In 

Mat1a KO this situation is aggravated, since SAMe-dependent 

PC synthesis via PE methylation is decreased, the PC/PE ratio 

reduced and mitochondrial FA oxidation impaired. To put a 

brake to this drain of PC molecules to LD, FA are rerouted in 

Mat1a KO mice liver to other catabolic (endoplasmic reticulum 

and peroxisome oxidation) and biosynthetic (ceramides 

synthesis) pathways, causing oxidative stress, inflammation 

and fibrosis. SAMe treatment for two months in 8-9 month 

old Mat1a KO mice ameliorated mitochondrial dysfunction 

(reduces membrane depolarization, improves Phb1 and Mcj 

expression, and increases SAMe transport to mitochondria) 

improving FA oxidation efficiency (FA and acylcarnitine levels 

decrease), which results in a drastic reduction in TG accumulation. 

SAMe treatment in Mat1a KO mice resulted in more PC 

available for proper membrane function, improving liver lipid 

homeostasis, histology (H&E, Sudan red, Sirius red) and liver 

injury (ALT, AST). 

Disclosures: 

The following authors have nothing to disclose: David Fernández, Cristina 

Alonso, Sebastiaan van Liempd, Marta Varela, Nicolás Navasa, Ibon Martinez-Arranz, 

Patricia Aspichueta, Ana M. Aransay, Juan Anguita, Juan M. Falcon-Perez, 

Maria L. Martinez-Chantar, Shelly C. Lu, Jose M. Mato 

954 

Angiogenesis in the pathogenesis of non-alcoholic fatty 

liver disease: a differential gene expression study 

Savneet Kaur 1 , Sebastian Vlaic 2 , Reinhard Guthke 2 , Rania Dayoub 

3 , Mohsin Hasan 4 , Hamda Siddiqui 1 , Thomas S. Weiss 3 , Shiv 

K. Sarin 4 ; 1 Gautam Buddha University, Greater Noida, India; 

2 Leibniz Institute for Natural Product Research and Infection Biology, 

Jena, Germany; 3 University children hospital (KUNO), University 

of Regensburg, Regensburg, Germany; 4 Institute of liver and 

biliary sciences, New Delhi, India 

Background: Nonalcoholic fatty liver disease (NAFLD) is a progressive 

liver disease that ranges from simple steatosis to the 

most severe form, nonalcoholic steatohepatitis (NASH). The 

onset of a chronic inflammatory reaction marks the progression 

from simple steatosis to NASH and the expansion of adipose 

tissue is strongly associated with neo-angiogenesis. Henceforth, 

in the current study, we investigated correlations between 

changes in the expression of lipid metabolism associated genes 

and genes which are associated with angiogenesis. Methods: 

Liver tissues were obtained from healthy controls, patients with 

simple liver steatosis, and with non-alcoholic steatohepatitis 

(NASH). Using Affymetrix Gene chips, differentially expressed 

genes (DEGs) were identified in 21 human liver tissue samples 

including healthy controls (n= 7), patients with simple liver 

steatosis (n= 7), and with NASH (n= 8). DEGs were identified 

using the R-package limma with a false discovery rate (FDR)-adjusted 

p-value of 0.1. Among all the DEGs, a subset of the 

DEGs associated with lipid metabolism and angiogenesis were 

selected. Further, potential key regulatory factors associated 

with both lipid metabolism and angiogenesis were analyzed 

to study relationships between these two pathways using a 

network inference approach. Results: Among the angiogenic 

genes, the genes associated with the chemokine signaling 

pathway and extracellular matrix proteins (ECM) were significantly 

upregulated in patients with NASH as compared to the 

patients with simple liver steatosis and healthy controls. Among 

the genes associated with lipid metabolism, genes associated 

with arachidonic acid metabolism and lipid biosynthesis were 

differentially expressed in NASH patients as compared to the 

patients with simple steatosis and controls. The network inference 

approach identified two genes, endothelin 1 (EDN1) and 

nuclear receptor, NR2F2 as key regulatory genes associated 

with both lipid metabolism and neo-vessel formation. The major 

targets of these genes included cyclooxygenase-2 (COX-2), 

chemokine receptor 4 (CXCR4), matrix metalloproteinases 9 

and 2 (MMP-9 and MMP-2). The analysis also inferred a novel 

positive correlation between COX-2 and MMP2, suggesting 

an important link between accumulation of MMPs and prostaglandin 

synthesis by COX-2. Conclusion: The study highlights 

the relationships between neovessel formation and lipid metabolism 

in pathogenesis of NASH. Further analysis on a larger 

cohort of patients and animal studies will shed light on how 

does angiogenesis leads to dysregulated lipid metabolism or 

vice-a-versa in NASH. 

Disclosures: 

The following authors have nothing to disclose: Savneet Kaur, Sebastian Vlaic, 

Reinhard Guthke, Rania Dayoub, Mohsin Hasan, Hamda Siddiqui, Thomas S. 

Weiss, Shiv K. Sarin 

955 

Glucagon-like peptide-1 potently attenuates steatohepatitis 

and liver fibrosis by regulating liver macrophage 

infiltration, activation and polarization 

Xiaoyu Wang 1 , Shih-Yen Weng 1 , Yong Ook Kim 1 , Olena Molokanova 

1 , Thomas Klein 2 , Detlef Schuppan 1,3 ; 1 Institute of Translational 

Immunology, Mainz, Germany; 2 Boehringer Ingelheim 

Pharma, Biberach an der Riss, Germany; 3 Division of Gastroenterology, 

Boston, MA 

Background and aims: Glucagon-like peptide-1 (GLP-1) 

improves insulin sensitivity via enhanced glucose-dependent 

insulin secretion, inhibition of glucagon release, and delayed 

gastric emptying following its release into the circulation from 

the gut. We aimed to explore the utility of the long-acting 

GLP-1 receptor agonist Bydureon (BY) to address both inflammation 

and fibrosis in models of NASH (nonalcoholic steatohepatitis) 

and biliary fibrosis. Methods: BY was administered 

twice weekly by subcutaneous injection of 0.4 or 2 mg/kg 

to Mdr2KO mice, and to C57BL/6 mice fed a methionine 

and choline deficient (MCD) diet for 4 weeks. Inflammation, 

fibrosis, steatohepatitis and especially macrophage function 

and polarization were assessed. Results: In both the MCD 

and Mdr2KO model, BY significantly decreased serum liver 

enzymes, liver collagen content, and fibrosis and inflammation 

related transcripts and protein levels (e.g., Col1a1, α-Sma, 

CD68, CCL3, and TNFα), while it increased the (anti-inflammatory) 

macrophage markers Arg1 and Ym1. BY treatment also 

reduced the IHC expression of collagen type III, CD68, F4/80 

and caspase3. In FACS analysis MCD livers showed no difference 

in CD11c + cells in all the groups, while the total percentage 

of F4/80 + macrophages was significantly decreased on 

BY treatment, indicating a direct effect of BY on macrophages 

but no significant impact on hepatic DC populations. More-


over, BY induced a significant decrease of liver CD11b + Ly6C hi 

inflammatory/fibrogenic myeloid cells and accompanied by 

a massive increase of hepatic CD11b + Ly6C lo cells. BY also 

reduced the CD68 + total macrophages and increased anti-inflammatory 

Ym1 + macrophages in Epididymal tissue and 

liver. Overall BY suppressed JNK phosphorylation and Erk1/2 

expression to inhibited fibrosis and inflammation. Conclusions: 

We show that GLP-1 exerts a direct beneficial effect on liver 

macrophages which is accompanied by significant effects on 

inflammation fibrosis and steatosis in models of biliary fibrosis 

and hepatic lipoapoptosis/NASH. Here it interferes with 

JNK signaling. Our data support further clinical evaluation of 

the utility of GLP-1R agonists for the treatment of patients with 

NASH and liver fibrosis. 

Disclosures: 

Thomas Klein - Employment: Boehringer Ingelheim 

Detlef Schuppan - Consulting: Boehringer Ingelheim, Conatus, GLG, Merck, 

Mitsubishi-Tanabe, Takeda, Silence, Glenmark, Isis; Grant/Research Support: 


The following authors have nothing to disclose: Xiaoyu Wang, Shih-Yen Weng, 

Yong Ook Kim, Olena Molokanova 

Disclosures: 






The following authors have nothing to disclose: Rohini Mehta, Michael J. Estep, 

Gary Bratthauer, Fanny Monge, Thomas Jeffers 

956 

Hepatic Estrogen Receptor Negatively Correlates with 

Fibrosis Only in Male Patients with Non-Alcoholic Steatohepatitis 

(NASH) 

Rohini Mehta 2 , Michael J. Estep 2 , Gary Bratthauer 2 , Fanny 

Monge 1 , Thomas Jeffers 2 , Zachary D. Goodman 1 , Zobair M. 

Younossi 2,1 ; 1 Center For Liver Disease, Department of Medicine, 

Inova Fairfax Hospital, Falls Church, VA; 2 Betty and Guy 

Beatty Center for Integrated Research, Inova Health Systems, Falls 

Church, VA 

Background and Aim: Evidence suggests that the pathogenesis 

of hepatic fibrosis may be influenced by gender and could 

potentially be related to estrogen. Our aim was to examine the 

relationship of estrogen receptor alpha (ERa) positive hepatocyte 

nuclei to the degree of hepatic fibrosis among male and 

female NASH patients. Methods: After informed consent, liver 

biopsies and clinical data were obtained from patients with 

biopsy-proven NASH (N=32; BMI=45.59 ± 14.65; Male 

Gender=50%; Age=45.16 ± 11.03). For immunohistochemistry, 

paraffin-embedded liver tissue (5uM) was de-paraffinized, 

rehydrated. The Anti-Estrogen Receptor alpha (clone ID5; 

AbCam) primary antibody (1:100 dilution) was applied to 

the sections. Visual staining was obtained using DAKO EnVision 

Dual link system. Peroxidase activity was visualized with 

3,3’-diaminobenzidine (DAB). Sections were lightly counterstained 

with hematoxylin. Ratio of positive hepatocyte nuclei to 

total hepatocyte nuclei were counted (Cell Sens,Olympus software) 

in three microscopic areas (60X) for each sample. Group 

comparisons by Mann-whitney analysis and Spearman’s correlation 

analysis were carried out. A p-value of ≤0.05 was 

considered to be statistically significant. Results: ER expression 

was detected in 75% (N=24) of patients. ER staining ratio 

(positive hepatocyte nuclei to total hepatocyte nuclei) ranged 

from 0%-85% in the entire population. Females had a wider 

range and a broader central tendency of ER positive hepatocytes 

(mean=31.62±27.88%, range: 0%-85.22%,) compared 

to males (mean=43.67±19.18, 1.62% - 68.57%, p>0.05). 

Males with an ER staining ratio >50% were associated with 

lower pericellular fibrosis (r=-0.51, p=0.04). However, this 

relationship was not detected in women. Conclusion: Hepatic 

ERa expression is negatively associated with degree of pericellular 

fibrosis in men but not women with NASH. Further studies 

are needed to establish differential role of ERa in males and 

females. 

957 

Liver-Directed Allosteric Inhibitors of Acetyl-CoA Carboxylase 

Reduce Hepatic Steatosis and Improve Dyslipidemia 

in Diet-Induced Obese Rat Models and Reduce 

Inflammation and Fibrosis in a Cirrhotic Rat Model 

Geraldine Harriman 1 , Danielle K. DePeralta 2 , Omeed Moaven 2 , 

Lan Wei 2 , Jeremy R. Greenwood 3 , Sathesh P. Bhat 3 , Kenneth K. 

Tanabe 2 , Bryan C. Fuchs 2 , William Westlin 1 , H. James Harwood 1 , 

Rosana Kapeller 1 ; 1 Nimbus Therapeutics, Cambridge, MA; 2 Massachusets 

General Hospital, Boston, MA; 3 Schrodinger, New York, 

NY 

Background: Non-alcoholic steatohepatitis (NASH) affects 16 

million people in the US and is projected to surpass hepatitis 

C as the leading cause of liver transplant by 2020. To date 

there are no approved drugs to treat NASH. It has been proposed 

that increased hepatic steatosis can lead to lipotoxicity 

and inflammation that drive development of NASH, fibrosis, 

cirrhosis, and HCC. The rate-limiting enzyme in fatty acid (FA) 

synthesis and oxidation is Acetyl-CoA Carboxylase (ACC). 

Pharmacologic inhibition of ACC presents an attractive mode 

for reducing de novo lipogenesis and enhancing FA oxidation. 

Using state-of-the-art structure-based drug design, we identified 

a unique series of allosteric ACC inhibitors that bind to its ‘BC’ 

domain, blocking enzymatic activity. Here we present data 

on the impact of liver-directed, selective ACC inhibition on 

the pathological processes involved in the progression from 

hepatic steatosis to NASH and cirrhosis. Methods: Liver-selective 

allosteric ACC inhibitors, ND-630 and ND-654, were 

evaluated to determine the effects of ACC inhibition on the 

specific pathologies associated with NASH and HCC including 

hepatic de novo lipogenesis, inflammation, and fibrosis. 

ND-630 and ND-654 inhibit HepG2 FASyn with EC 50 

of 

8.7nM and 4nM, and hepatic FASyn in vivo with ED 50 

of 

0.14mg/kg and 0.3mg/kg, respectively. We tested the efficacy 

of these compounds in a high sucrose diet-induce obesity 

(HS DIO) rat model and in a diethylnitrosamine (DEN)-induced 

rat model of cirrhosis and HCC. Results: Inhibition of ACC 

with ND-630 in a rat model of HS DIO led to potent and dose 

responsive decrease in plasma triglycerides, cholesterol, free 

fatty acids, and basal insulin and reduced hepatic triglyceride 

levels with maximal inhibition on Day 14 at 10 mg/kg. Inhibition 

of hepatic ACC by ND-654 produced a dose-dependent 

decrease in liver fibrosis, as determined by Sirius Red stain,


and a decrease in mRNA for markers of inflammation and stellate 

cell activation CD-68 and α-SMA, respectively, in a DEN 

model of cirrhosis. Furthermore, ND-654 markedly decreased, 

in a dose dependent manner, DEN-induced protein expression 

of inflammatory cytokines including MCP1, TNFα and LIF, and 

fibrotic markers such as TIMP1 and FGF21 and other markers 

including Serpin E1, Leptin and Resistin. Conclusions: These 

data demonstrate that liver-directed inhibition of ACC can 

impact multiple steps in the pathogenesis of fatty liver disease 

to NASH and cirrhosis . 

Disclosures: 

Geraldine Harriman - Employment: Nimbus Discovery 

Jeremy R. Greenwood - Employment: Schrodinger Inc.; Patent Held/Filed: Nimbus 

Therapeutics; Stock Shareholder: Schrodinger Inc. 

Kenneth K. Tanabe - Patent Held/Filed: EGF SNP and risk for HCC, EGFR inhibition 

and HCC, gene signature for prognosis in cirrhosis 

Bryan C. Fuchs - Consulting: Collagen Medical; Grant/Research Support: Nimbus 

Therapeutics 

Rosana Kapeller - Employment: Nimbus Therapeutics; Stock Shareholder: Aileron 

Therapeutics 

The following authors have nothing to disclose: Danielle K. DePeralta, Omeed 

Moaven, Lan Wei, Sathesh P. Bhat, William Westlin, H. James Harwood 

958 

Metformin targets a phosphoSTAT3-miRNAs pathway 

to inhibit lipid droplets accumulation and intracellular 

inflammation in vitro and in vivo 

Natalia Pediconi 1 , Silvia Di Cocco 3 , Silvia Piconese 3 , Federica 

Mori 4 , Laura Belloni 2 , Abigail D. Nunn 2 , Tullio Scopigno 2 , Vincenzo 

Barnaba 3 , Giovanni Blandino 4 , Sabrina Strano 4 , Massimo 

Levrero 3,2 ; 1 Dept of Molecular Medicine, Sapienza University, 

Rome, Italy; 2 Center for Life NanoScience (CNLS), IIT-Sapienza, 

Rome, Italy; 3 Dept of Internal Medicine - DMISM, Sapienza University, 

Rome, Italy; 4 Translational Oncogenomic Unit, Regina Elena 

Cancer Institute, Rome, Italy 

Background and aim: Accumulation of triglyceride-containing 

lipid droplets (LDs) within hepatocytes in NAFLD patients is a 

potentially reversible process, although sustained activation of 

inflammatory signaling pathways leads to non-alcoholic steatohepatitis 

(NASH) that can evolve into cirrhosis and HCC. 

The prevalence of NAFLD is increased with aging, however 

the molecular mechanism for aging-induced fatty liver remains 

poorly understood. Here we investigated the role of a phosphoSTAT3-miRNAs 

pathway in LD accumulation and the 

activation of intracellular inflammatory pathways in vescicular 

steatosis. Methods: DMSO-differentiated human non-transformed 

HepaRG cells treated with oleic acid were used as a 

cellular model of vescicolar steatosis. C57BL/6 mice treated or 

not with metformin for 78 weeks were used as an in vivo model 

of aging-induced fatty liver. Results: dHepaRG cells treated 

with oleic acid show: a) an accumulation of intracellular lipids 

with an increase of both total cellular lipid volume and lipid 

droplets number, as evaluated by picosecond CARS (coherent 

anti-stoke Raman spettroscopy) mycroscopy; b) the generation 

of reactive oxygen species (ROS); c) deregulated lipid metabolism 

and liver-specific genes, including PDK4, PLIN4, SLC2A1, 

ALB and ALDOB; d) activation of an intracellular inflammatory 

response, with the upregulation of NFkB and IFN-a regulated 

genes and the activation of the phosphoSTAT3/IL6 pathway. 

Treatment of oleate-exposed HepaRG with the S3I STAT3 

inhibitor reduces bothe tryglicerides and ROS accumulation 

and inhibits phosphoSTAT3 accumulation. We also found that 

several STAT3/IL6 responsive miRNAs, including miR21 and 

miR24, are upregulated after lipid overload, paralleling STAT3 

activation. Chromatin immuno-precipitation (ChIP) experiments 

showed that the oleate-dependent transcriptional deregulation 

of these miRNAs correlates with phosphoSTAT3 bound to their 

promoters. Treatment with S3I inhibits both oleate-dependent 

miR21 and miR24 upregulation and phosphoSTAT3 binding to 

their promoter. Treatment with metformin, an AMPK activator 

widely used as anti-diabetic drug and known to reduce liver 

steatosis in vivo, reduces phosphoSTAT3 accumulation, inhibits 

miRNAs upregulation and reduces trygliceride accumulation in 

fatty HepaRG. Importantly, a chronic 78 weeks treatment with 

metformin prevents aging-induced steatosis in C57BL/6 mice 

and reduces both liver and serum miR21 levels. Conclusions: 

We show that a phosphoSTAT3-miRNAs intracellular inflammatory 

pathway amenable to therapeutic targeting by metformin 

is activated in response to lipid accumulation in differentiated 

hepatocytes in vitro and in vivo. 

Disclosures: 

Massimo Levrero - Advisory Committees or Review Panels: BMS, Jansen, Gilead, 

Tekmira, Galapagos, Medimmune; Speaking and Teaching: MSD, Roche 

The following authors have nothing to disclose: Natalia Pediconi, Silvia Di 

Cocco, Silvia Piconese, Federica Mori, Laura Belloni, Abigail D. Nunn, Tullio 

Scopigno, Vincenzo Barnaba, Giovanni Blandino, Sabrina Strano 

959 

Exacerbation of non-alcoholic fatty liver disease 

(NAFLD) in mice with myeloid cell-specific deletion of 

FXR 

Rachel McMahan 1 , Cara Porsche 1 , Moshe Levi 2 , Hugo R. Rosen 1 ; 

1 Gastroenterology, University of Colorado, Aurora, CO; 2 Renal 

Diseases & Hypertension, University of Colorado, Aurora, CO 

Background: Nonalcoholic fatty liver disease (NAFLD) is estimated 

to affect a third of the U.S. population. The progression 

from benign steatosis to steatohepatitis in NAFLD has been 

linked to increased recruitment of pro-inflammatory monocytes 

and activation of liver macrophages. We have previously 

shown that activation of the nuclear bile acid receptor FXR can 

improve disease in murine models of NALFD and that in vitro 

activation of FXR in macrophages inhibits pro-inflammatory 

cytokine production. However, FXR is also expressed in other 

cell types including hepatocytes, and the protective effects of 

FXR may be through non-macrophage FXR signaling. Therefore, 

we aimed to determine the specific role of macrophage 

FXR expression in NALFD. Methods: To evaluate to role of 

macrophage expression of FXR in NAFLD, we used the Cre- 

Lox system to create myeloid-cell specific targeted mutants of 

the FXR gene (FXR fl/fl LyzMCre). FXR fl/fl LyzMCre or littermate 

controls were fed a western or control diet for 3 months and 

NAFLD severity was determined by histological analysis, 

hepatic gene expression and flow cytometric analysis of intrahepatic 

immune cells isolated by collagenase and mechanical 

digestion. Results: Loss of FXR expression in myeloid cells led to 

a more severe histological phenotype with increased macrovesicular 

steatosis and inflammatory foci compared to littermate 

controls. In addition, western diet feeding increased hepatic 

expression of pro-inflammatory genes (CCL2, TNF-alpha) and 

pro-fibrotic genes (TIMP-1) in FXR fl/fl LyzMCre mice. Myeloid 

cell deletion of FXR also led to increased recruitment of intrahepatic 

pro-inflammatory monocytes and an increase in IL-17 

producing T cells. Conclusions: Myeloid cell-specific deletion of 

FXR leads to increased liver inflammation in a murine model of 

NAFLD. Targeting FXR in macrophages may be important for 

inhibiting hepatic inflammation and NAFLD development. 

Disclosures: 



The following authors have nothing to disclose: Rachel McMahan, Cara Porsche, 

Hugo R. Rosen


960 

miRNA-339-3p is increased in human and experimental 

models of NAFLD and targeted by TUDCA in insulin-resistant 

muscle cells 

André L. Simão 2 , Marta B. Afonso 2 , Pedro M. Rodrigues 2 , Andreia 

J. Amaral 5 , Margarida Gama-Carvalho 5 , Pedro M. Borralho 1 , Mariana 

V. Machado 3 , Helena Cortez-Pinto 3,4 , Cecília M. Rodrigues 1 , 

Rui E. Castro 1 ; 1 iMed.ULisboa & Dep. of Biochemistry and Human 

Biology, Faculty of Pharmacy, University of Lisbon, Lisboa, Portugal; 

2 iMed.ULisboa, Faculty of Pharmacy, University of Lisbon, 

Lisbon, Portugal; 3 Gastrenterology, Hospital Santa Maria, Lisbon, 

Portugal; 4 Faculty of Medicine, University of Lisbon, Lisbon, Portugal; 

5 BioISI– Biosystems & Integrative Sciences Institute, Faculty of 

Sciences, University of Lisbon, Lisbon, Portugal 

Intramyocellular lipid deposition associates with insulin resistance 

(IR), constituting a key pathophysiological event in human 

non-alcoholic fatty liver disease (NAFLD). microRNAs (miRNA/ 

miRs) play also a functional role in NAFLD both in the liver 

and in other metabolic tissues, including the muscle. Finally, 

tauroursodeoxycholic acid (TUDCA) is cytoprotective in both 

liver and muscle cells, in part by modulating insulin-signaling 

pathways. Our aims were to evaluate the functional role of 

muscle miR-339-3p in human and animal models of NAFLD, as 

well as in insulin-resistant C2C12 muscle cells, in the presence 

or absence of TUDCA. Skeletal muscle biopsies were obtained 

from morbid obese NAFLD patients undergoing bariatric surgery. 

Muscle and plasma RNA were extracted with Trizol (Life 

Technologies) and the miRCURY kit (Exiqon), respectively. 

Human muscle RNA was run in TaqMan MicroRNA arrays 

for global miRNA expression analysis. C57BL6 mice were fed 

either a standard or a fast food (FF) diet for 25 weeks. C2C12 

muscle cells were incubated with or without 200 or 400 mM 

palmitic acid (PA), +- 100 mM of TUDCA, for evaluation of the 

insulin-signaling pathway, mitochondrial function and overall 

cellular toxicity. Six miRNAs, including miR-339-3p, a regulator 

of glucose synthesis, were found to progressively and 

significantly increase from steatosis to NASH (at least p




Frank Tacke - Advisory Committees or Review Panels: Tobira; Grant/Research 

Support: Novartis, Noxxon; Speaking and Teaching: BMS, Gilead, Falk, MSD, 

Janssen, Abbvie 

The following authors have nothing to disclose: Robert Schierwagen, Lara Maybüchen, 

Sebastian Zimmer, Kanishka Hittatiya, Christer Baeck, Sabine Klein, 

Frank E. Uschner, Winfried Reul, Peter Boor, Georg Nickenig, Jogchum Plat, 

Dieter Luetjohann, Jonel Trebicka 

962 

TRPV4 deficiency enhances TLR4 recruitment to lipid 

rafts exhibiting exacerbated stellate cell activation and 

fibrosis in experimental nonalcoholic steatohepatitis 

Suvarthi Das 1 , Ratanesh K. Seth 1 , Varun Chandrashekaran 1 , Firas 

Alhasson 1 , Diptadip Dattaroy 1 , Gregory A. Michelotti 2 , Prakash 

Nagarkatti 3 , Phillip D. Bell 4 , Wolfgang B. Liedtke 2 , Anna Diehl 2 , 

Saurabh Chatterjee 1 , Mitzi Nagarkatti 3 ; 1 Environmental Health 

Sciences, University of South Carolina, Columbia, SC; 2 Duke University, 

Durham, NC; 3 University of South Carolina, Columbia, 

NC; 4 Medical University of South Carolina, Charleston, SC 

Transient receptor potential vanilloid channel 4 (TRPV4) is a 

nonselective cation channel that confers sensitivity to extracellular 

osmolarity and regulates calcium inflow. We have observed 

previously that TRPV4 protein levels are significantly higher in 

NASH rodent models and human NASH patients. For the present 

study we aimed to investigate the role of TRPV4 in stellate 

cell activation and fibrosis in a methionine choline deficient 

(MCD) diet-induced mouse model of NASH. Liver homogenates 

from mice fed with MCD diet for 8 weeks (8 to 16 weeks 

of age), were used for qRTPCR and Western Blots. Paraffin 

embedded liver sections from the same mice were used for 

immunostaining and histopathological analysis. The Results 

showed that TRPV4 deficiency in mice fed with an MCD diet 

for 8 weeks showed significantly higher -SMA immunoreactivity 

in livers followed by significantly higher macro and micro 

vesicular fibrosis (Picrosirius Red staining) when compared to 

MCD wild type mice. TRPV4 deficiency led to inflammasome 

activation as evidenced by significantly high Apoptosis-associated 

speck-like protein containing a CARD (ASC II), cleaved 

IL-1β and cleaved Caspase-1 immunoreactivity when assessed 

by immunoblots as compared to wild type mice. Mice deficient 

in TRPV4 showed higher mRNA expressions of MCP-1 and 

IL-1β in the livers as compared to wildtype mice, an observation 

that correlated well with histopathology where increased 

infiltrating leukocytes were present in the liver. Since NASH 

progression and fibrosis were associated with increased lipid 

raft recruitment of TLR4 primarily mediated by NADPH oxidase, 

we studied whether TRPV4 deficiency could lead to 

enhanced NADPH oxidase driven TLR4 recruitment to lipid 

rafts in these mice. Results showed that there was a significant 

increase in NADPH oxidase activation as evidenced by colocalization 

of membrane gp91phox and p47phox subunits in 

TRPV4 knockout MCD mice as compared to wild type MCD 

mice. This correlated well with a significant increase in the TLR4 

colocalization in flotillin containing rafts in the cell membranes. 

These dual labeled liver sections were imaged using immunofluorescence 

microscopy. The TLR4 raft recruitment was also 

accompanied by significant elevation in TLR4 protein in the 

TRPV4 deficient mice. Finally, we conclude and propose that 

MCD diet feeding might generate severe oxidative stress which 

possibly leads to TRPV4 led calcium influx and might aid in 

preservation of membrane integrity while lack of TRPV4 causes 

severe loss of membrane structure causing leakage of damage 

associated molecular patterns into cells causing enhanced 

inflammation, stellate cell activation and fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Suvarthi Das, Ratanesh K. Seth, 

Varun Chandrashekaran, Firas Alhasson, Diptadip Dattaroy, Gregory A. Michelotti, 

Prakash Nagarkatti, Phillip D. Bell, Wolfgang B. Liedtke, Anna Diehl, 

Saurabh Chatterjee, Mitzi Nagarkatti 

963 

β 7 

-Integrins and MAdCAM-1 have opposing functions in 

development and progression of Non-alcoholic steatohepatitis 

(NASH) 

Hannah K. Drescher 1 , Angela Schippers 2 , Thomas Clahsen 2 , 

Hacer Sahin 1 , Norbert Wagner 2 , Christian Trautwein 1 , Konrad L. 

Streetz 1 , Daniela C. Kroy 1 ; 1 Department of Internal Medicine III, 

University Hospital, RWTH Aachen, Germany, Aachen, Germany; 

2 Department of Pediatrics, University Hospital RWTH Aachen, 

Aachen, Germany 

Introduction: Non-alcoholic steatohepatitis (NASH) is the third 

most common reason for liver transplantations in developing 

countries and one of the fastest growing medical problems. 

Aim: The significance of infiltrating leukocytes and how they 

affect NASH development and progression remains unclear. 

Therefore, this study investigates the role of the homing receptor 

pair MAdCAM-1/β 7 

in two different mouse NASH-models. 

Methods: Constitutive β 7 

-Integrin and MAdCAM-1 knockout 

mice were fed either MCDdiet (methionine and choline deficient) 

for 4 weeks or HF-diet (high fat) for 26 weeks. Results: 

After 4 weeks of MCD treatment β 7 

-deficient animals displayed 

earlier and faster progressing steatosis reflected by significant 

histomorphological changes while MAdCAM-1-KO mice 

showed an intact liver architecture, both compared to wildtype 

controls. Consistent with severe disease progression β 7 

-Integrin 

deficient animals showed an increased oxidative stress 

response (DHE (Dihydroethidium)-staining) with simultaneous 

down regulation of the expression of Treg markers (FoxP3, 

IL-2, TGFβ). In contrast, MAdCAM-1-KO mice displayed an 

up-regulation of anti-oxidative stress proteins involved and an 

enhanced number of infiltrating macrophages tending to provide 

a strengthening of the anti-inflammatory response. The 

β 7 

-KO group exhibited a stronger hepatic infiltration of inflammatory 

cells reflecting an earlier onset of NASH. Especially 

Th17 positive T cells were increased leading to elevated numbers 

of infiltrating neutrophils. Those changes finally resulted 

in an earlier and stronger collagen accumulation in these animals 

while MAdCAM-1-KO mice were protected from fibrosis 

progression. The results suggest a direct effect of the homing 

receptor pair MAdCAM-1/β 7 

on the gut-liver-axis in NASH 

development by an infiltration blockade of cells which have 

a positive immuno-modulatory effect on the liver. In a second 

model, better reflecting the metabolic changes during NASH 

development, HF-diet was fed to mice for 26 weeks. Preliminary 

results show comparable results to those in the MCD 

model, namely a significantly increased bodyweight and a 

worsened glucose tolerance in β 7 

-KO animals indicating the 

occurrence of the metabolic syndrome in this group compared 

to MAdCAM-1-KO and WT. Conclusion: MAdCAM-1 and β 7 

Integrin 

deficiency leads to opposing effects in the MCD and the 

HF model, with protection in MAdCAM-1-KO animals and a 

more severe phenotype and significantly stronger fibrosis progression 

in β 7 

-Integrin-KO mice. Therefore, the interaction of 

β 7 

Integrins and their receptor MAdCAM-1 provide a potential 

novel target for therapeutic interventions during NASH development. 

Disclosures: 


Speaking and Teaching: Roche, BMS, Roche, BMS


The following authors have nothing to disclose: Hannah K. Drescher, Angela 

Schippers, Thomas Clahsen, Hacer Sahin, Norbert Wagner, Konrad L. Streetz, 

Daniela C. Kroy 

964 

Association between nonalcoholic fatty liver disease 

and bone mineral density: a systematic review and 

meta-analysis 

Sikarin Upala 1,3 , Veeravich Jaruvongvanich 4,5 , Karn Wijarnpreecha 

1,2 , Anawin Sanguankeo 1,3 ; 1 Department of Internal Medicine, 

Bassett Medical Center and Columbia University College of Physicians 

and Surgeons, Cooperstown, NY; 2 Department of Physiology, 

Cardiac Electrophysiology Research and Training Center, 

Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand; 

3 Department of Preventive and Social Medicine, Faculty of 

Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand; 

4 Department of Internal Medicine, University of Hawaii, Honolulu, 

HI; 5 Department of Internal Medicine, Faculty of Medicine, Chulalongkorn 

University, Bangkok, Thailand 

Purpose: Several major risk factors for osteoporosis have been 

identified. One of these risk factors is chronic inflammation. 

Several recent studies have supported the association between 

low bone mineral density (BMD) and nonalcoholic fatty liver 

disease (NAFLD), which comprises a spectrum of disorders 

involving liver inflammation. However, conflicting evidence 

regarding this association has been obtained thus far. We 

therefore conducted a meta-analysis of observational studies 

to show the association between NAFLD and BMD. Methods: 

The Cochrane Central Register of Controlled Trials, Cochrane 

Library, Medline, and Embase were searched from database 

inception to November 2014 for all observational studies evaluating 

the association between NAFLD or nonalcoholic steatohepatitis 

(NASH) and bone mass, BMD, or osteoporosis. All 

patients were ≥18 years of age and had no other cause of liver 

disease, osteoporosis, or pathological bone disease at baseline. 

Risk factors were NAFLD and NASH; control subjects were 

individuals without NAFLD. Results: Eleven articles underwent 

full-length review. Data were extracted from 5 cross-sectional 

studies involving 1,276 participants; 638 had NAFLD. The 

main meta-analysis showed no significant difference in BMD 

between patients with fatty liver disease and controls. This may 

be explained by the confounding effects of body mass index on 

BMD in patients with NAFLD. Among all variables analyzed, 

body mass index had the strongest and most significant predictive 

effect on the difference in BMD. Conclusions: Controversy 

exists regarding the effect of BMD on NAFLD. Further studies 

are required to fully show this relationship. 

965 

Aliskiren reduces liver and epididymal fat steatosis and 

increases skeletal muscle insulin sensitivity in high-fat 

diet-fed mice 

Kuei-Chuan Lee, Yun-Cheng Hsieh, Ying-Ying Yang, Che-Chang 

Chan, Yi-Hsiang Huang, Han-Chieh Lin; Taipei Veterans General 

hospital, Taipei, Taiwan 

Aliskiren has been found to reduce chronic injury and steatosis 

in the liver of methionine-choline-deficient (MCD) diet-fed 

mice. This study investigated whether aliskiren has an anti-steatotic 

effect in HFD-fed mice, which are more relevant to human 

patients with non-alcoholic fatty liver disease than MCD mice. 

Mice fed with 4-week normal chow or HFD randomly received 

aliskiren (50 mg/kg/day) or vehicle via osmotic minipumps for 

further 4 weeks. Aliskiren reduced systemic insulin resistance, 

hepatic steatosis, epididymal fat mass and increased gastrocnemius 

muscle glucose transporter type 4 levels with lower 

tissue angiotensin II levels in the HFD-fed mice. In addition, aliskiren 

lowered nuclear peroxisome proliferator-activated receptor 

gamma and its down-signaling molecules and increased 

cytochrome P450 4A14 and carnitine palmitoyltransferase 1A 

(CPT1a) in liver. In epididymal fat, aliskiren inhibited expressions 

of lipogenic genes, leading to decrease in fat mass, body 

weight, and serum levels of leptin and free fatty acid. Notably, 

in the gastrocnemius muscle, aliskiren increased phosphorylation 

of insulin receptor substrate 1 and Akt. Based on these 

beneficial effects on liver, peripheral fat and skeletal muscle, 

aliskiren is a promising therapeutic agent for patients with 

NAFLD. 

Disclosures: 

The following authors have nothing to disclose: Kuei-Chuan Lee, Yun-Cheng 

Hsieh, Ying-Ying Yang, Che-Chang Chan, Yi-Hsiang Huang, Han-Chieh Lin 

966 

A DPP-4 inhibitor, sitagliptin, can be preventative 

drug for the acceleration of the development in NAFLD 

caused by lipopolysaccharides through inhibiting 

expression of MyD88 and NFκB. 

Keisuke Yokohama 2,1 , Shinya Fukunishi 1 , Sanomura Makoto 2 , 

Akira Asai 1 , Yasuhiro Tsuds 1 , Kazuhide Higuchi 1 ; 1 2nd Department 

of internal medicine, Osaka Medical College, Osaka, Japan; 

2 Gastroenterology, Hokusetsu general hospital, Osaka, Japan 

Disclosures: 

The following authors have nothing to disclose: Sikarin Upala, Veeravich Jaruvongvanich, 

Karn Wijarnpreecha, Anawin Sanguankeo 

Aims: Nonalcoholic fatty liver disease (NAFLD) is a chronic 

liver disease, and commonly observed in patients with obesity 

or type 2 diabetes mellitus (T2DM). Characterized by metabolic 

syndrome, hepatic steatosis, and liver inflammation, 

nonalcoholic steatohepatitis(NASH)is believed to be under 

the influence of the gut microflora. Some reports showed that


lipopolysaccharides (LPS) contained in normal portal blood 

are one of the factors about the development of NASH. Two 

major pathway for LPS have been suggested in some studies, 

which are referred to as MyD88-dependent and –independent 

pathways. We aimed to determine the effects of sitagliptin on 

the development of LPS-induced NAFLD in ob/ob mice with 

diet-induced obesity and T2DM. Methods: Five-week-old male 

ob/ob mice, which develop T2DM and NAFLD by taking a 

high-carbohydrate diet (HCD), were divided into a group in 

which a HCD was given for 8 weeks and treated with LPS 

(n=6) as controls (Control group), and another in which a HCD 

added with 0.0018% sitagliptin was given for 8 weeks and 

treated with LPS (n=6) (Sitagliptin group). Results: Histological 

examination demonstrated that this protocol induced steatosis 

in the livers of two group mice, however severe fatty droplets 

were significantly observed in Control group, compared 

with Sitagliptin group. Sitagliptin group showed a decrease 

in serum ALT and triglyceride, in addition, hepatic mRNA 

expression levels of SREBP-1c, FAS and peroxisome PPARγ 

were decreased in Sitagliptin group. Furthermore, Sitagliptin 

inhibits the mRNA expression levels of proinflammatory gene 

such as TNF-α. Interestingly, hepatic mRNA expression levels of 

MyD88 and NFκB were decreased in Sitagliptin group. These 

finding coincided with histological findings and the results of 

biochemical examination. Conclusion: Our findings indicated 

that sitagliptin could be preventative drug for the development 

of LPS-induced NAFLD and MyD88-dependent pathway. 

Disclosures: 

The following authors have nothing to disclose: Keisuke Yokohama, Shinya 

Fukunishi, Sanomura Makoto, Akira Asai, Yasuhiro Tsuds, Kazuhide Higuchi 

967 

DSS colitis has tumorinogenesis and fibrogenesis in choline-deficiency 

and high-fat diet induced NASH mouse 

model 

Koichi Achiwa, Masatoshi Ishigami, Yoji Ishizu, Teiji Kuzuya, 

Takashi Honda, Kazuhiko Hayashi, Yoshiki Hirooka, Hidemi Goto; 

Gastroenterology and Hepatology, Nagoya Univercity Graduate 


Background and Aim Non-alcoholic steatohepatitis (NASH) 

patients progress to liver cirrhosis and even hepatocellular carcinoma(HCC). 

Several line of evidence indicated that accumulation 

of lipopolysaccharide (LPS) plays a contributory role in 

the pathogenesis HCC by eliciting proinflammatory response in 

the liver. Moreover, it has been reported that dextran sodium 

sulfate (DSS) induced colitis mouse model fed high fat diet 

increased portal LPS and developed hepatic inflammation and 

fibrosis. However, diet induced NASH model with carcinogenesis 

require over 50 weeks or more. The purpose of this study 

was to create a novel tumorigenesis in diet induced NASH 

with DSS administration. Methods 20-week-age C57BL/6 

mice were fed choline-deficiency and high-fat diet (CDHF) for 

4 and 12 weeks. DSS group was fed CDHF and 1% DSS in 

the drinking water intermittently. On the other hand control 

group was fed CDHF and free water supply. We evaluated the 

liver tumorigenesis. Hepatic lipid was measured and inflammatory 

and fibrosis factors and histological changes were evaluated. 

Results Exposure of DSS led to increase mucosal change 

such as crypt loss and increase of inflammatory cells in colon. 

Accordingly, the histological score significantly increased in 

DSS group than control group. But a few colon tissues were 

increased inflammatory cells at 12 week in control group. 

CDHF led to slightly cause intestinal inflammation. Portal LPS 

levels were increased in DSS group comparing with control 

group at 4 week, whereas there was no significant difference 

in both groups at 12week. Hepatic triglyceride, total and free 

cholesterol were increased in the DSS group comparing with 

control group at 4 week, whereas there was no significant 

difference in both groups at 12 week. Both groups showed 

severe macrovesicular steatosis in liver histology. Serum total 

bilirubin levels were significantly higher in DSS group than that 

in control group. Serum transaminase levels are higher in DSS 

group than that in control group. A large number of lobular 

inflammatory cells were seen in DSS group. The image analysis 

showed significant elevation of F4/80 staining positive area in 

DSS group comparing with control group. Fibrosis is increased 

in DSS group compared to control group. Liver tumors were 

developed in the DSS group of 12-week, while tumor was not 

found in the DSS group of 4-week and in the control group. 

One of tumors showed highly differentiated hepatocellular carcinoma. 

Conclusions DSS administration developed liver tumors 

in CDHF induced NASH mouse at the short term. Induction of 

intestinal inflammation in NASH may promote hepatic tumorigenesis. 

Disclosures: 



The following authors have nothing to disclose: Koichi Achiwa, Masatoshi Ishigami, 

Yoji Ishizu, Teiji Kuzuya, Takashi Honda, Kazuhiko Hayashi, Yoshiki 

Hirooka 

968 

Inhibition of Mitophagy in Non-alcoholic Steatohepatitis 

lead to Apoptosis and Inflammation 

Muhammad Sohail 1 , Seong-Jun Kim 2 , Aleem Siddiqui 2 ; 1 GASTRO- 

ENTEROLOGY, university of California, San Diego, CA; 2 Infectious 

Disease, University of California, San Diego, CA 

Background: Fatty liver is benign but it can progress to Non-Alcoholic 

Steatohepatitis (NASH) and liver cirrhosis. Mitochondria 

are dynamic organelles that constantly undergo fission, 

fusion, and mitophagy to facilitate mitochondrial quality control, 

which is crucial for maintaining cell viability and bioenergetics. 

Mitochondrial fission/fragmentation is mediated 

by recruitment of cytosolic Drp1 to the mitochondria forming 

spirals that constrict both the inner and outer mitochondrial 

membranes. Drp1 recruitment to mitochondria is regulated by 

phosphorylation and dephosphorylation of respective serine 

residues by putative kinases and phosphatases. Mitochondrial 

dynamics is tightly regulated in response to alterations 

in cellular physiology and its dysfunction occurs in NASH. 

Methods Palmitic acid (PA) is able to reproduce in vitro model 

for fatty liver disease. To investigate the mitochondrial fission 

and mitophagy, the human hepatoma Huh7 cells cultured in 

the presence or absence of PA were observed by confocal 

microscope. PA-induced stimulation and mitochondrial translocation 

of dynamin-related protein 1 (Drp1) and Parkin, two 

key factors of mitochondrial fission and mitophagy, were analyzed 

by Western blot using purified mitochondria fraction 

and confocal microscope. The liver tissues extracted from highfat 

diet-fed C57BL/6 mice and biopsy samples from Human 

NASH patients were also used for analysis of Drp1 and Parkin 

activation. Small interfering RNA-mediated Drp1 and Parkin 

silencing strategy was further employed for further analysis of 

PA-induced mitochondrial fission and mitophagy and alteration 

in inflammatory response. Results: PA stimulated Drp1 gene 

expression and induced mitochondrial translocation of Drp1 

via promoting both its phosphorylation (Ser616) and dephosphorylation 

(Ser637) followed by mitochondrial fission. These 

results were confirmed in liver tissue extracted from high-fat 

diet-fed C57BL/6 mice and also in human NASH liver biopsy 

tissue. While PA induced Parkin gene expression, mitophagy


was not observed. PA also induce PINK1 cleavage, implying 

the possible inflammatory and apoptosis signaling. Using specific 

protease inhibitor that cleaves PINK1 in inner mitochondrial 

membrane, we were able to block its cleavage which 

also leads to the inhibition of apoptosis. As mitophagy does not 

occur which is important mitochondrial quality control, this lead 

to surplus damaged mitochondria and subsequent apoptosis 

of cell further triggering inflammation. Conclusion: Fatty acids 

induce aberrant mitochondrial dynamics involving mitochondrial 

fission, caused apoptosis but does not lead to mitophagy. 

Disclosures: 

The following authors have nothing to disclose: Muhammad Sohail, Seong-Jun 

Kim, Aleem Siddiqui 

Figure: NFκB inhibition by AP inhibits IL-1β expression induced by 

LPS. (A) AP treatment inhibited NFκB activation by LPS in HepG2 

cells, evaluated through a reporter gene assay. (B) NFκB inhibition 

by AP treatment inhibited IL1β induction by LPS in Palmitic 

acid-treated HepG2 cells. (**p≤0.01, ***p ≤ 0.001). 

969 

NFkB inhibition by Andrographolide ameliorates 

inflammation and fibrogenesis through inflammasome 

substrate depletion in experimental Non-Alcoholic Steatohepatitis 

(NASH) 

Daniel Cabrera 1,2 , Alexander Wree 3 , Davide Povero 3 , Nancy 

Solis 1 , Margarita Pizarro 1 , Pamela Rojas de Santiago 1 , Javiera 

Torres 7 , Han Moshage 5 , Claudio Cabello-Verrugio 4 , Ariel E. 

Feldstein 3 , Enrique Brandan 6 , Marco Arrese 1 ; 1 Departamento de 

Gastroenterología, Facultad de Medicina Pontificia Universidad 

Católica de Chile, Santiago, Chile; 2 Departamento de Ciencias 

Químico-Biológicas, Universidad Bernardo O Higgins, Santiago, 

Chile; 3 Department of Pediatrics, University of California San 

Diego, San Diego, CA; 4 Departamento de Ciencias Biológicas, 

Universidad Andres Bello, Santiago, Chile; 5 Department of Gastroenterology 

and Hepatology, University of Groningen, Santiago, 

Chile; 6 Departamenti de Biologia Celular y Molecular, Pontificia 

Universidad Católica de Chile, Santiago, Chile; 7 Departamento 

de Anatomia Patologica, Pontificia Universidad Católica de Chile, 

Santiago, Chile 

Background: Options to treat NASH are limited. Andrographolide 

(AP), the bioactive component of Andrographis paniculata, 

has potent anti-inflammatory activity related to NFκB 

inhibition. NFkB is a crucial factor in Interleukin-1β (IL-1β) 

expression, the main substrate for the inflammasome. Aim: To 

evaluate the effects of AP in experimental NASH and its influence 

in inflammasome activity. Methods: C57bl6 mice were 

fed a choline-deficient-amino-acid–defined (CDAA) diet (22 

weeks) with or without AP (1 mg/Kg, 3 times/week, intraperitoneally). 

Serum levels of alanine aminotransferase (ALT) and 

hepatic steatosis, inflammation, and fibrosis were assessed. 

Hepatic triglyceride content (HTC) and hepatic mRNA levels 

of selected pro-inflammatory and pro-fibrotic genes as well as 

those related to inflammasome were also measured. Direct AP 

effect on IL-1β expression and inflammasome activity was evaluated 

in HepG2 cells loaded with a mixture of FFAs during 24h. 

Results: AP decreased HTC and attenuated hepatic inflammation 

and fibrosis in CDAA-fed mice reducing serum ALT activity 

and hepatic collagen deposition. AP treatment was associated 

with a strong reduction in hepatic macrophage infiltration 

and of hepatic mRNA levels of both pro-inflammatory and 

pro-fibrotic genes. In addition, mice treated with AP showed 

reduced expression of inflammasome genes (-60% reduction in 

inflammasome adaptor ASC hepatic mRNA levels, and -40% 

reduction in NLRP3 hepatic mRNA levels). Finally, AP inhibited 

IL-1β expression induced by LPS through NFκB inhibition in fat 

laden HepG2 cells (figure). Conclusion: AP administration prevented 

liver damage in NASH. Inflammasome inactivation by 

NFκB-dependent mechanism involving IL-1β expression inhibition 

is likely involved in the therapeutic effects of AP. (Fondecyt 

1150327 to M.A. and FONDECYT PD3140396 to D.C.) 

Disclosures: 

The following authors have nothing to disclose: Daniel Cabrera, Alexander 

Wree, Davide Povero, Nancy Solis, Margarita Pizarro, Pamela Rojas de Santiago, 

Javiera Torres, Han Moshage, Claudio Cabello-Verrugio, Ariel E. Feldstein, 

Enrique Brandan, Marco Arrese 

970 

Reduction of Hepatic 27-Hydroxycholesterol in Steatohepatitis 

Model Mice with Insulin Resistance 

Sho-ichiro Yara 1 , Tadashi Ikegami 1 , Akira Honda 1,2 , Teruo 

Miyazaki 2 , Masashi Murakami 1 , Junichi Iwamoto 1 , Yasushi Matsuzaki 

1 ; 1 Division of Gastroenterology and Hepatology, Tokyo 

Medical University Ibaraki Medical Center, Ami, Inashiki, Japan; 

2 Collaborative Research Center, Tokyo Medical University Ibaraki 

Medical Center, Ami, Inashiki, Japan 

BACKGROUND: 27-hydroxycholesterol (HC), one of the major 

oxysterol found in the human circulation, is produced by the 

mitochondrial enzyme CYP27A1. Previous report demonstrated 

that hepatic inflammation was increased by genetic deletion of 

CYP27A1 and decreased by 27HC treatment in high fat diet 

(HFD) fed steatotic mice, however, direct cause of dysregulation 

of CYP27A1 and 27HC in steatohepatitis is unknown. 

AIM: The current study was undertaken to examine the change 

of hepatic OS levels in the Stelic Animal Model (STAM) mice, 

a validated and widely used as NASH-derived HCC model. 

METHODS: Extracted sterols in the liver tissue homogenates 

were determined by LC-MS/MS. RESULTS: In STAM mice, IR 

was induced by injection of streptozotocin at the birth period. 

Four weeks after injection, feeding of high-fat diet was started, 

and given over time. At week 20, STAM mice developed hepatocellular 

carcinoma (HCC) whereas HCCs were not found in 

control (CTL) as well as mice only fed with HFD. (1) Concentration 

of hepatic cholesterol (CHOL): Hepatic CHOL concentrations 

were kept in the narrow range (2.5 to 2.95 mg/mg wet 

weight liver (ww)) by 12 weeks old either in CTL, HFD fed, or 

STAM mice, but significantly elevated at 20 weeks in HFD fed 

mice (4.7 mg/mg ww) and STAM mice (4.2 mg/mg ww) compared 

to CTL (2.2 mg/mg ww). (2) Concentration of hepatic 

bile acids (BAs): In HFD fed mice, hepatic BA concentrations 

(111.1 to 120.8 nmol/g ww) were significantly lower than CTL 

(177.4 to 205.6 nmol/g ww) over time, suggesting enhanced 

BA excretion from liver under the condition with higher hepatic 

CHOL level in the absence with IR. In STAM mice, hepatic BA 

concentrations were maintained in a similar level of HFD at 

earlier phase (4 to 8 wk old), but those were significantly elevated 

at later phase (12 to 20wk old)(215.0 nmol/g ww at 12 

wk, 289.0 nmol/g ww at 20wk). (3) Concentration of hepatic 

OS: Among major OS tested (24S, 25epo-HC, 22RHC, 24HC, 

25HC, 27HC, 4βHC), only 27HC was significantly lower in 

STAM mice (0.388 to 0.579 ng/mg ww) compared to either


of CTL (0.606 to 0.942 ng/mg ww) or HFD (0.628 to 0.755 

ng/mg ww) over time. CONCLUSION: Reduced hepatic 27HC 

concentration observed in this model, is supportive the hypothesis 

proposed by previous studies. Enhancement of the reduction 

of hepatic 27HC concentration in the presence of IR suggested 

a linkage between IR and dysregulation of CYP27A1. 

Disclosures: 

The following authors have nothing to disclose: Sho-ichiro Yara, Tadashi Ikegami, 

Akira Honda, Teruo Miyazaki, Masashi Murakami, Junichi Iwamoto, Yasushi 

Matsuzaki 

971 

Role of Fcgamma receptors in experimental Non-Alcoholic 

Steatohepatitis 

Daniel Cabrera 1,3 , Evelyn Jara 2 , Ricardo Cruz 1 , Natalia 

Muñoz-Durango 2 , Pamela Rojas de Santiago 1 , Alexis Kalergis 2 , 

Marco Arrese 1 ; 1 Departamento de Gastroenterología, Facultad 

de Medicina, Pontificia Universidad Católica de Chile, Santiago, 

Chile; 2 Departamento de Genética Molecular y Microbiología, 

Facultad de Ciencias Biológicas, Pontificia Universidad Católica 

de Chile, Santiago, Chile; 3 Departamento de Ciencias Químico-Biológicas, 

Universidad Bernardo O Higgins, Santiago, Chile 

Background: The role of innate immunity in Non-alcoholic steatohepatitis 

(NASH) development is emergent. Different populations 

of innate immune cells are present in the liver controlling 

tissue cytokine production through Fc gamma receptors (FcγRs), 

which are receptors for the Fc region of IgG antibodies. FcγRs 

cell-type specifically interact with various other receptors for 

selective amplification or inhibition of particular cytokines. 

Aim: To evaluate the role of the inhibitory (FcγRIIB) and activatory 

(FcγRIII) receptors in NASH pathogenesis. Methods: 

Wild Type, FcγRIIb(-/-) and FcγRIII(-/-) mice were fed a methionine-choline 

deficient (MCD) diet for 5 weeks. Liver injury was 

assessed by measuring serum levels of alanine aminotransferase 

(ALT) and histologically. Hepatic triglyceride content (HTC) 

and hepatic mRNA levels of selected pro-inflammatory (TNF-α, 

IFN-γ, IL-1β, etc.), pro-fibrotic (TGF-β, CTGF, Collagen-1, etc.) 

and inflammasome (ASC, NLRP3, Caspase-1, etc.) genes were 

also assessed. Serum pro-inflammatory cytokine levels (TNF-α, 

IFN-γ, etc.) were determined by cromatographic bead assay 

(CBA). By flow cytometry was evaluated different populations 

of inflammatory cells infiltrated in the liver such as dendritic, 

neutrophils, macrophages and lymphocytes. Results: FcγRIIb(-/-) 

MCD-fed mice developed a more robust hepatic inflammatory 

(decreased hepatic expression of TNFα and other cytokines) 

and fibrotic response (decreased hepatic expression of collagen 

I) in comparison with WT MCD-fed mice with no differences 

in HTC (Figure 1 A and B). No differences were found 

in liver cell populations of lymphoid and myeloid lineages. 

The main finding in FcγRIII(-/-) MCD-fed mice was a significant 

reduction in histological steatosis and HTC. (see figure 1C, 

below) likely related to reduced interleukin-10 production. Liver 

lymphoid and myeloid cell populations remained unchanged in 

these mice. Conclusion: Our results suggest and important role 

of the FcγRs in NASH development. While the absence of Fcγ− 

RIIb seems to promote NASH induction, the absence of FcγRIII 

strongly reduces liver steatosis. This is the first report that shows 

a direct role of FcγRs in the pathogenesis of NASH.(Grant 

support: FONDECYT 1150327 to M.A., PD3140396 to D.A.) 

Disclosures: 

The following authors have nothing to disclose: Daniel Cabrera, Evelyn Jara, 

Ricardo Cruz, Natalia Muñoz-Durango, Pamela Rojas de Santiago, Alexis Kalergis, 

Marco Arrese 

972 

Regulation of Hepatocellular Fatty Acid Uptake in 

Mouse Models of Fatty Liver Disease With and Without 

Functional Leptin Signalling: Roles of NfKB and 

Srebp-1c 

Fengxia Ge, Jose L. Walewski, Paul D. Berk; Medicine, Columbia 

University Medical Center, New York, NY 

The specific processes leading to increased hepatic triglycerides 

(TGs) in mouse models of hepatic steatosis (HS) due 

to EtOH consumption, high fat diets (HFDs), or obesity mutations 

remain uncertain. This study focuses on regulation by 5 

transcription factors (Nfb, Srebp-1c, AMPK, PPARα, PPARγ) of 

the 7 most-studied hepatic LCFA transporters (FABPpm, CD36, 

FATP1, FATP2, FATP4, FATP5, & Caveolin-1 [CAV-1]) and 

enzymes of LCFA synthesis (SCD-1, FASN) in mice with HS 

from various causes. Methods: Seven groups of 20 wk old 

male mice (n=8/group; Jackson Labs) were studied: C57BL/6J 

controls (C); similar mice with HS from 12 wks of a high fat diet 

(HFD) or 10, 14, or 18% EtOH in drinking water (E10, E14, 

E18); & genetically obese mice lacking leptin (ob/ob) or its 

receptor (db/db). Hepatocyte [ 3 H]-oleate uptake was assayed 

by rapid filtration; Vmax for saturable uptake was computed 

with SAAM II. Gene expression for the 5 transcription factors, 

7 transporters, & for key enzymes of LCFA synthesis was 

assayed by qRT-PCR. Hepatic TG was measured biochemically. 

Expression ratios for transcription factors and transporter 

genes were compared with one another, with Vmax, & with 

hepatic TG. Results: [1] LCFA uptake Vmax was increased & 

highly correlated with hepatic TG in all groups except ob/ob & 

db/db. [2] Increased Vmax & hepatic TG in the EtOH & HFD 

groups correlated best with increased expression of genes for 

at least one & often multiple transporters. [3] Despite variable 

expression of single transporter genes in individual E & HFD 

groups, the mean expression ratio for FABPpm, FATPs 1,2,4,& 

5, & CD36 in each group was highly correlated with Vmax, 

hepatic TG, and expression of transcription factor genes. [4] 

Of the transcription factors, SREBP-1c (r = 0.99) and NfKB (r 

= 0.94) were by far the most closely correlated with Vmax. [5] 

Increased hepatic TGs in ob/ob & db/db mice did not relate 

to hepatic LCFA uptake, but instead were highly correlated 

with increased expression of LCFA synthetic enzymes (SCD-1, 

FASN). Conclusions: [1] The processes underlying increased 

hepatic TG are different in mice with vs without functional leptin 

signaling. Increased LCFA uptake is the principal cause of HS 

in the former, but increased LCFA synthesis predominates in 

the latter. [2] Regulation of LCFA transporter expression & participation 

of individual transporters in LCFA uptake are more 

complex than previously believed. [3] Correlations between 

transcription factor expression & mean expression of multiple 

transporter genes suggests possible regulatory interaction, and 

may support reports postulating that a complex of FABPpm, 

CAV-1, CD36 & FATP4 mediates hepatic LCFA uptake.


Disclosures: 

The following authors have nothing to disclose: Fengxia Ge, Jose L. Walewski, 

Paul D. Berk 

973 

Role of the receptor tyrosine kinase Mer in the development 

of fibrosis in NAFLD 

Giovanni Di Maira 1 , Salvatore Petta 2 , Andrea Cappon 1 , Elisa 

Vivoli 1 , Luca Valenti 3 , Paola Dongiovanni 3 , Elisabetta Bugianesi 4 , 

Vito Di Marco 2 , Fabio Marra 1 ; 1 University of Florence, Florence, 

Italy; 2 University of Palermo, Palermo, Italy; 3 University of Milan, 

Milan, Italy; 4 University of Turin, Turin, Italy 

Background/aims: Non-alcoholic fatty liver disease (NAFLD) 

describes a spectrum of conditions ranging from steatosis to 

non-alcoholic steatohepatitis (NASH), which may progress 

to fibrosis, cirrhosis and hepatocellular carcinoma. Several 

genetic factors involved in NAFLD severity and progression 

have been identified through genome-wide association studies 

(GWAS) but their biologic significance and sites of action is 

still unclear. Mer (MerTK) is a receptor tyrosine kinase with 

oncogenic properties, over-expressed or activated in various 

malignancies. In a recent GWAS, a SNP of the non-coding 

region of MerTK (rs4374383 G>A) has been associated with 

fibrosis severity in patients with chronic hepatitis C, but currently 

there is no evidence of the involvement of this factor 

in the progression of NAFLD. Aim of this study was to assess 

the possible role of MerTK in the fibrogenic progression of 

NAFLD. Methods: Genetic analyses were performed on specimens 

from patients who underwent liver biopsy for suspected 

NASH without severe obesity. For murine models of fibrogenesis, 

C57BL6/J mice were treated with CCl4 for 6 weeks, and 

Balb/C mice were fed with a methionine and choline deficient 

(MCD) diet for 8 weeks. Biopsy samples from patients with 

NAFLD and different stages of fibrosis were also analyzed. 

Intrahepatic gene expression was measured by qPCR. Human 

HSC were isolated from normal liver tissue and cultured on 

plastic. UNC569 was used to inhibit MerTK activity. Results: 

The MerTK rs4374383 AA polymorphism, which is linked with 

a decreased expression of MerTK, was associated with a lower 

prevalence of clinically significant fibrosis in patients with 

NAFLD. In murine models of hepatic fibrogenesis, an increased 

hepatic expression of MerTK was observed. In human hepatic 

stellate cells (HSC) Mertk was found to be highly expressed 

at the gene and protein levels. Stimulation of MerTK with the 

cognate ligand, GAS6, resulted in time-dependent activation of 

ERK1/2 and in stimulation of cell migration. Moreover, pharmacological 

inhibition of MerTK with UNC569 reduced HSC 

survival activating apoptotic pathways. Finally, patients with 

NAFLD and severe fibrosis had significantly higher intrahepatic 

expression of MerTK than those with mild or no fibrosis. 

Conclusion: These data indicate the novel role of MerTK as a 

modulator of fibrogenesis in NAFLD and identify HSC as a 

cellular target of this kinase. 

Disclosures: 

Fabio Marra - Advisory Committees or Review Panels: Abbvie; Consulting: Bayer 

Healthcare; Grant/Research Support: ViiV 

The following authors have nothing to disclose: Giovanni Di Maira, Salvatore 

Petta, Andrea Cappon, Elisa Vivoli, Luca Valenti, Paola Dongiovanni, Elisabetta 

Bugianesi, Vito Di Marco 

974 

GFT505 (ELAFIBRANOR) prevents nonalcoholic steatohepatitis 

(NASH), hepatic fibrosis and hepatocarcinoma 

in a new disease model 

Benoit Noel 1 , Geraldine Rigou 1 , Nathalie Degallaix 1 , Valérie 

Daix 1 , Linda Cambula 1 , Alice Roudot 1 , Rémy Hanf 1 , Dean W. 

Hum 1 , Bart Staels 2 , Robert Walczak 1 ; 1 Genfit SA, Loos, France; 

2 INSERM UMR1011, Institut Pasteur, Lille, France 

Background: NAFLD is present in 20-30% of the adult population 

in developed countries. NASH, cirrhosis and hepatocellular 

carcinoma will develop in a substantial proportion of 

people with NAFLD. Recently, the phase 2B GOLDEN-505 trial 

has demonstrated the efficacy of GFT505 in reversing NASH 

in patients with advanced disease. In the present study we 

have assessed the efficacy of GFT505 in preventing NASH, 

liver fibrosis and hepatocarcinoma development in a new rat 

model. Methods: Steatohepatitis, NASH and hepatocarcinoma 

were induced by feeding Wistar rats with a choline-deficient 

L-amino-acid-defined-diet (CDAA) that was supplemented with 

1% cholesterol. In the intervention group, animals also received 

GFT505 (10mg/kg/day PO) for the entire study period. 

NASH, fibrosis and hepatocarcinoma development were evaluated 

by histology. Additional biochemical and molecular 

analyses were also performed on different relevant biomarkers. 

Results: CDAA diet administration in Wistar rats causes 

liver transaminase elevation and fibrosing steatohepatitis that is 

similar to human NASH pathology. In this study the supplementation 

of the CDAA diet with 1% cholesterol led to increased 

NASH incidence (92% vs 58%) and to fibrosis aggravation as 

evidenced by higher prevalence of cirrhotic animals (42% vs. 

8%). NASH development was significantly attenuated in rats on 

CDAA diet that were also administered GFT505. Consistently, 

hepatic inflammation, hepatocyte ballooning, oxidative stress 

and plasma transaminase levels were decreased, whereas 

plasma levels of FGF-21 and β-hydroxybutyrate, as well as 

the expression of fatty acid oxidation genes in the liver were 

increased in rats that received GFT505. Hepatic collagen was 

lower, fibrosis and cirrhosis development were all significantly 

prevented by GFT505 administration on CDAA diet. Finally, 

GFT505 administration completely prevented pre-neoplastic 

lesion development in rats exposed to CDAA diets as revealed 

by the GST-P staining. Conclusion: GFT505 prevented NASH, 

fibrosis, cirrhosis and pre-neoplastic lesion development in this 

novel NASH model. Since the effect of GFT505 on hepatic 

steatosis was rather modest in this particular model, this study 

demonstrates that the antifibrotic activity of GFT505 can be 

dissociated from its hypolipidemic efficacy. 

Disclosures: 

Benoit Noel - Employment: Genfit SA 

Nathalie Degallaix - Employment: GENFIT SA 

Linda Cambula - Employment: GENFIT SA 

Alice Roudot - Employment: GENFIT 

Rémy Hanf - Management Position: GENFIT 

Dean W. Hum - Management Position: Genfit 


Robert Walczak - Management Position: Genfit SA 

The following authors have nothing to disclose: Geraldine Rigou, Valérie Daix


975 

Free fatty acid treatment reduces BMP6 signalling in 

AML12 hepatocytes 

Nishreen Santrampurwala 1,2 , Kim Bridle 1,2 , Janske Reiling 1,3 , 

Laurence J. Britton 1,2 , Lesley-Anne Jaskowski 1,2 , Nathan Subramaniam 

1,4 , Darrell H. Crawford 1,2 ; 1 School of Medicine, The University 

of Queensland, Greenslopes, QLD, Australia; 2 Gallipoli 

Medical Research Foundation, Brisbane, QLD, Australia; 3 NUTRIM 

School of Nutrition and Translational Research in Metabolism, 

Maastricht University, Maastricht, Netherlands; 4 QIMR Berghofer 

Medical Research Institute, Brisbane, QLD, Australia 

Introduction: Individuals with non-alcoholic steatohepatitis 

(NASH) frequently have increased hepatic iron stores, which 

appear to potentiate liver injury. The underlying mechanism of 

iron loading in these patients however has remained elusive. 

Our group has previously shown a greater severity of injury in 

Hfe -/- mice fed a high calorie diet (HCD) compared to wild type 

controls. Despite iron loading, mice fed a HCD had markedly 

reduced expression of hepcidin (Hamp1), the master regulator 

of systemic iron homeostasis. BMP (bone morphogenetic 

protein)-SMAD signalling is known to be important in hepcidin 

induction; we hypothesised that BMP signalling is altered 

in a fatty acid-rich environment. We developed an in vitro 

model of NAFLD to examine the consequences of accumulated 

fatty acids on the BMP-SMAD signalling pathway. Methods: 

The mouse hepatocyte cell line (AML12) was serum-starved 

for 4 hours before treatment with free fatty acids (FFA; 2mM 

of oleate and palmitate in a 2 to 1 ratio), ferric ammonium 

citrate (Fe, 100μM) and/or BMP6 (50ng/ml) for 12-hours. 

RNA and protein were extracted for further downstream analysis. 

Results: Serum-starved AML12 cells treated with BMP6 

had increased expression of Hamp1, Smad7, Id1 and Atoh8 

indicating an intact BMP6 signalling pathway. Cells cultured 

with FFA however had a blunted response to BMP6 treatment. 

This was evident by the significant decrease in expression of 

Hamp1, Smad7, Id1 and Atoh8 (p ≤ 0.05) in cells treated with 

FFA despite BMP6 treatment. Additionally, we investigated the 

levels of phospho-SMAD1/5/8, a BMP6 signalling intermediary, 

and found that while phosphorylation of SMAD1/5/8 

was stimulated with BMP6 treatment the induction was significantly 

blunted with FFA treatment (p ≤ 0.001). Discussion: Iron 

loading in NAFLD has the potential to increase oxidative stress 

and lipid peroxidation, thus contributing to progressive liver 

disease. The mechanisms underlying iron loading in NAFLD 

are largely unknown and reduced BMP-SMAD signalling in 

response to FFA treatment represents a plausible mechanistic 

link, by decreasing hepcidin expression, thus facilitating duodenal 

iron absorption via ferroportin. 

Disclosures: 

Darrell H. Crawford - Advisory Committees or Review Panels: Roche Products 

Pty Ltd, Bristol Myers Squibb, Gilead Sciences, Novartis, MSD, Abbvie, Jansen; 

Consulting: Roche Products Pty Ltd; Grant/Research Support: Roche Products 

Pty Ltd; Speaking and Teaching: Roche Products Pty Ltd, Bristol Myers Squibb, 

Gilead Sciences, MSD 

The following authors have nothing to disclose: Nishreen Santrampurwala, Kim 

Bridle, Janske Reiling, Laurence J. Britton, Lesley-Anne Jaskowski, Nathan Subramaniam 

976 

Increased sensitivity and compromised time-dependent 

adaptation to methoxamine in a rat model of severe 

steatosis 

Wilhelmus J. Kwanten 1 , Paul Fransen 2 , Joris G. De Man 1 , Benedicte 

Y. De Winter 1 , Peter P. Michielsen 1,3 , Sven M. Francque 1,3 , 

Denise Van Der Graaff 1 , Michiel Landen 1 ; 1 Laboratory of Experimental 

Medicine and Pediatrics (LEMP) - Gastroenterology & 

Hepatology, University of Antwerp, Wilrijk, Belgium; 2 Laboratory 

of Physiopharmacology, University of Antwerp, Wilrijk, Belgium; 

3 Gastroenterology Hepatology, Antwerp University Hospital 

(UZA), Edegem, Belgium 

NAFLD causes important intrahepatic vascular alterations and 

an increased portal blood pressure, even before the development 

of inflammation or fibrosis. Intrahepatic endothelial 

dysfunction and a decreased sensitivity to vasoconstriction to 

the α1-adrenoceptor agonist methoxamine (Mx) were reported. 

AIM To study the underlying mechanisms of the reported hyporesponsiveness 

to Mx and potential time-dependent effects. 

METHODS Male Wistar rats were fed a methione-choline deficient 

diet (MCDD) to induce steatosis or control diet (CD) diet 

for 4 weeks (n=4-5/group). Intrahepatic vascular resistance 

was studied by in situ isolated liver perfusion (flow: 10-50 

ml/min). Dose-response curves to Mx were constructed evaluating 

intrahepatic vascular tone. Finally, intrahepatic vascular 

tone was assessed as a function of time at a fixed dose 

(at Emax, 10-4 M Mx) and flow (30ml/min). RESULTS Basal 

intrahepatic resistance didn’t differ between steatotic and 

control rats, nor was the response to increasing flow rates. 

Dose-response curves to Mx showed a significantly increased 

sensitivity to Mx in steatosis compared to controls (-LogEC50: 

5.55 vs 5.20; p60min: 19.24>11.25mmHg). Steatotic 

livers showed a similar initial contraction, while time-dependent 

decrease was delayed and attenuated (5>60min: 

19.03>16.88mmHg)(fig 1). Significance was achieved after 

20min Mx. When expressed as percentage decrease compared 

to the level after 5min Mx, significance was already 

achieved after 10min. CONCLUSION Our results show a significant 

and time-dependent adaptation of intrahepatic vascular 

tone to Mx in control rats. This phenomenon should be taken 

into account when interpreting experiments with Mx pre-constriction. 

Secondly, although we didn’t observe an increased 

intrahepatic vascular resistance, response to Mx was significantly 

altered in steatosis, with a significant hyperresponsiviness 

and impaired time-dependent adaptation. 

Disclosures: 

The following authors have nothing to disclose: Wilhelmus J. Kwanten, Paul 

Fransen, Joris G. De Man, Benedicte Y. De Winter, Peter P. Michielsen, Sven M. 

Francque, Denise Van Der Graaff, Michiel Landen


977 

NorUDCA reduces liver injury and improves glucose 

sensitivity in a mouse model of obesity and steatosis. 

Daniel Steinacher 1 , Thierry Claudel 1 , Tatjana Stojakovic 2 , Michael 

Trauner 1 ; 1 Gastroenterology and Hepatology, Medical University 

of Vienna, Vienna, Austria; 2 Clinical Institute of Medical and 

Chemical Laboratory Diagnostics, Medical University of Graz, 

Graz, Austria 

Background: Non-alcoholic fatty liver disease ranges from simple 

benign steatosis to the more severe forms of non-alcoholic 

steatohepatitis (NASH), cirrhosis and ultimately hepatocellular 

carcinoma. Currently there is no satisfactory drug therapy 

against NASH available. The leptin deficient mouse (ob/ob) 

mimics metabolic syndrome and suffers from hyperinsulinemia, 

insulin resistance, hyperlipidemia, hepatic steatosis and inflammation. 

Bile acids (BA) are not only fat-absorption facilitators 

but can also regulate metabolic processes directly. Currently, 

ursodeoxycholic acid (UDCA) is the only BA used as a therapeutic 

in humans, but has limited efficacy in NASH. NorUDCA 

is a side-chained shortened derivative of UDCA improving liver 

injury in mouse models of cholestatic liver and bile duct injury. 

Therefore, we aimed to explore whether NorUDCA improves 

liver damage and has beneficial metabolic effects in ob/ob 

mice. Material & Methods: ob/ob mice received either a diet 

supplemented with 0.5% NorUDCA or chow diet for 6 weeks. 

Intraperitoneal glucose and insulin tolerance tests (IPGTT, IPITT) 

were performed at week 5 and 6. Serum and urine biochemistry, 

liver and fat histology were analyzed, gene and protein 

expressions of key markers of inflammation, lipid and glucose 

metabolism were assessed by RT-PCR and Western Blotting. 

Results: Mice treated with NorUDCA showed a significant 

reduction in AST, ALT and AP serum levels (p


mice compared with wild-type mice displayed elevated plasma 

ALT (150 +/-11.94 vs 108+/-4), and more fat accumulation 

in the liver. These data suggest that BHB production by the 

beta-oxidation pathway, and sensing by GPR019a, protects 

from alcohol induced acute liver injury. 

Disclosures: 

The following authors have nothing to disclose: Yonglin Chen, Irma Garcia-Martinez, 

Xinshou Ouyang, Rafaz Hoque, Wajahat Z. Mehal 

980 

Interferon lambda 3 (IL28B) and interferon lambda 4 

genotypes and resistance-associated variants in HCV 

genotype 1 and 3 infected patients 

Kai-Henrik Peiffer, Lisa Sommer, Simone Susser, Julia Dietz, Dany 

Perner, Caterina Berkowski, Stefan Zeuzem, Christoph Sarrazin; J. 

W. Goethe University, Medizinische Klinik 1, Frankfurt am Main, 

Germany 

Background and aims: In the era of pegylated interferon 

(Peg-IFN) single nucleotide polymorphisms (SNPs) in the interferon 

lambda 3 (IFN-L 3, also known as IL28B) and interferon 

lambda 4 genes (IFN-L4) were strong predictors for achieving 

SVR in patients with hepatitis C virus (HCV) infection. For many 

direct acting antiviral (DAA) combination regimens only weak 

or no correlation with IFN-L SNPs was observed. However, little 

is known about a potential selection of resistance-associated 

variants (RAVs) by IFN-L genotypes, which may influence virologic 

treatment response. This study aims to analyze the prevalence 

of rare and common RAVs in NS3, NS5A and NS5B 

to currently approved DAAs in a large European population 

in correlation to IFN-L3 and IFN-L4 genotypes. Materials and 

methods: Samples of 633 patients chronically infected with 

HCV genotype 1a (n=259), 1b (n=323) and 3 (n=51) were 

genotyped for rs12979860 (IFN-L3, also known as IL28B) and 

ss469415590 (IFN-L4) by real-time PCR. RAVs in NS3, NS5A 

and NS5B were detected via population-based sequencing and 

correlated with the IFN-L SNPs. Results: No significant correlation 

was found between IFN-L genotypes and rare and common 

RAVs within NS3 and NS5B genes (i.e. Q80K, C316N and 

S556G/N/R). In contrast, the NS5A RAV Y93H was detected 

frequently in HCV genotype 1b (14%) and was significantly 

associated with the beneficial rs12979860 (IFN-L3) C/C- and 

the ss469415590 (IFN-L4) TT/TT-genotype (p=0.0005 and 

p=0.0016, respectively). Therefore, prevalence of the Y93H 

variant is much higher in IFN-L3 C/C (27%) and IFN-L4 TT/TT 

(28%) than in non-C/C (10%) and non-TT/TT (10%) genotyped 

patients respectively. In addition, Y93H was found to be significantly 

associated with higher viral loads independent from 

the IFN-L genotype (p=0.000936 in all HCV genotypes). Summary: 

The major NS5A RAV Y93H is significantly associated 

with the presence of beneficial IFN-L3 and IFN-L4 SNPs and a 

high baseline viral load in HCV genotype 1 infected patients, 

which may explain a lack or inverse correlation of treatment 

response with IFN-L genotype in NS5A inhibitor containing 

IFN-free regimens and might be of clinical interest for consideration 

of IFN-free treatment options for patients with known 

IFN-L3 or IFN-L4 status. In addition Y93H was found to be 

associated with a higher viral load independent from the IFN-L 

genotype, which may partially explain in HCV genotype 1b the 

known paradox phenomenon of higher viral loads in patients 

with the beneficial IFN-L3 genotype. 

Disclosures: 









The following authors have nothing to disclose: Kai-Henrik Peiffer, Lisa Sommer, 

Simone Susser, Julia Dietz, Dany Perner, Caterina Berkowski 

981 

MicroRNA-130a inhibit HCV production through regulating 

host immune responses and lipid metabolism 

Xiaoqiong Duan 1 , Limin Chen 1,2 ; 1 Institute of Blood Transfusion,Chinese 

Academy of Medical Sciences (CAMS) /Peking 

Union Medical College (PUMC), Chengdu, China; 2 Toronto General 

Research Institute,University of Toronto, Toronto, ON, Canada 

Background & Aims: MicroRNAs (MiRNAs)a class of small 

non-coding RNAs that regulate messenger RNA (mRNA) 

expression, play pivotal role in the regulation of viral infections. 

In our previous study, we demonstrated that over-expression 

of miR-130a inhibited HCV replication by restoring 

the host innate immune response. But the detailed mechanism 

remains unclear. In this study, we aimed to identify and validate 

the potential target genes of miR-130a to explore the 

possible regulation mechanism between miR-130a and host 

immune responses in the context of HCV infection. Methods: 

Four algorithms, miRanda, TargetScan, PITA and RNAhybrid 

were used to predict the potential targets of miR-130a. To validate 

the target genes, MiR-130a mimic were transfected to 

Huh7.5.1 cells for 48h and total RNAs were extracted and 

reverse-transcribed to cDNA. Quantitative real-time PCR and 

western blot were used to measure target gene expressions 

with or without miR-130a overexpression. Results: Three potential 

target genes pyruvate kinase (PKLR), interleukin 18 binding 

protein (IL18BP) and low density lipoprotein receptor (LDLR) 

were identified by software algorithms and validated by qRT- 

PCR. miR-130a over-expression inhibited the expression levels 

of PKLR, IL18BP and LDLR significantly both mRNA and at protein 

levels. Functional analysis suggested that these genes are 

involved in lipid metabolism and host immune response. Quite 

interestingly, Cholesterol-25-hydroxylase (CH25H), an interferon-stimulated 

gene playing critical role in lipid metabolism 

and anti-HCV innate immunity, was stimulated by miR-130a. In 

addition, miR-130a overexpression increased the expression 

levels of interferon γ (IFNγ) and inhibited HCV entry, possibly 

through its targeting to IL18BP and LDLR, respectively. Conclusion: 

miR-130a inhibits HCV production through regulating 

host immune responses and lipid metabolism. Mir-130a may 

be developed as a potential drug candidate for the treatment 

of HCV infection.


Disclosures: 

The following authors have nothing to disclose: Xiaoqiong Duan, Limin Chen 

982 

Individualized DAA treatment duration for cure in 

patients with cirrhosis via modeling of early HCV kinetics 

Martina Gambato 3 , Laetitia Canini 1,4 , Sabela Lens 3 , Frederik 

Graw 5 , María-Carlota Londoño 3 , Susan L. Uprichard 1 , Zoe 

Mariño 3 , Enric Reverter Segura 3 , Concepció Bartres 3 , Patricia 

Gonzalez 3 , Scott Cotler 1 , Xavier Forns 3 , Harel Dahari 1,2 ; 1 The 

Program for Experimental & Theoretical Modeling, Department 

of Medicine, Division of Hepatology, Loyola University Medical 

Center, Maywood, IL; 2 Theoretical Biology & Biophysics Group, 

Los Alamos National Laboratory, Los Alamos, NM; 3 Liver Unit, 

Hospital Clínic, IDIBAPS and CIBEREHD, Barcelona, Spain; 4 Centre 

for Immunity, Infection and Evolution, University of Edinburgh, 

Edinburgh, United Kingdom; 5 Center for Modeling and Simulation 

in the Biosciences, BioQuant Center,, Heidelberg University, Heidelberg, 

Germany 

Background. While recent DAA-based regimens achieve SVR 

rates of over 90%, there is still substantial interest in optimizing 

length (and cost) of therapy, particularly in difficult-to-cure populations, 

such as those with cirrhosis. We aimed to investigate 

whether modelling of early HCV-RNA kinetics after therapy 

initiation could predict the minimal treatment duration needed 

to achieve SVR (cure). Methods. Fifty eight HCV patients who 

received DAA-based treatment for 12, 16 or 24 weeks at a 

single centre were included. HCV genotype, Child-Turcotte- 

Pugh [CTP], MELD, hepatic venous pressure gradient [HVPG], 

liver stiffness [LS], platelets count [PLT], albumin level [ALB]), 

IL28B, and treatment regimen were recorded. Viral load was 

assessed at baseline, at 4, 8 and 24 hours, at days 2, 3, 4, 

7, and at weeks 2, 3 and 4 after treatment initiation (or until 

target-not-detected (TND); LLOQ: HCV-RNA


The following authors have nothing to disclose: Donatella Palazzo, Elisa Biliotti, 

Francesca Tinti, Alessandra Bachetoni, Stefania Grieco, Andrea Cappoli, Raffaella 

Labriola, Paola Perinelli, Ilaria Umbro, Paola Rucci, Anna Paola Mitterhofer 

984 

Hepatitis C virus facilitates its persistent infection 

through ΔNp63-miR-181a-SIRT1/ DUSP6 signaling 

pathways 

Yun Zhou 1,2 , Guangyu Li 2,3 , Junping Ren 2,3 , Ying Zhang 1 , Jianqi 

Lian 1 , Changxing Huang 1 , Shunbin Ning 2,3 , Jonathan P. Moorman 

2,3 , Zhansheng Jia 1 , Zhi Q. Yao 2,3 ; 1 Department of Infectious 

Diseases, Tangdu Hospital, Fourth Military Medical University, 

Xi’an, China; 2 Department of Internal Medicine, Division of Infectious 

Diseases, James H. Quillen College of Medicine, East Tennessee 

State University, Johnson City, TN; 3 Center for Inflammation, 

Infectious Disease and Immunity, James H. Quillen College of Medicine, 

East Tennessee State University, Johnson city, TN 

T cells play a crucial role for viral clearance or persistence; 

however, the precise mechanisms that control their responses 

during viral infection remain largely unknown. MicroRNAs 

(miR) have been implicated as key regulators controlling 

diverse biological processes through posttranscriptional repression. 

Here we demonstrate that hepatitis C virus (HCV)-mediated 

regulation of ΔNp63 and miR-181a expression impairs 

CD4+ T cell responses to facilitate viral persistence via the 

dual specific phosphatase 6 (DUSP6) and SIRT1 signaling pathways. 

Specifically, a significant up-regulation of ΔNp63 leads 

to a decline of miR-181a expression, concomitant with over-expression 

of DUSP6 and SIRT1 in CD4+ T cells from chronically 

HCV-infected individuals compared to healthy subjects. 

The levels of ΔNp63 expression were found to be negatively 

associated with the levels of miR-181a, which in turn correlate 

to the expressions of DUSP6 and SIRT1 in these cells. Importantly, 

silencing ΔNp63/SIRT1 or reconstitution of miR-181a 

expression in CD4+ T cells leads to T cell senescence including 

reduced telomerase activity, shortened telomere length, 

increased senescence-associated β-galactosidaese (SA-β-gal) 

expression, and decreased EdU incorporation. Paradoxically, 

an increased IL-2 expression is observed in CD4+ T cells by 

these treatments, primarily due to a reduced frequencies of 

Foxp3+ regulatory T cells (Tregs) and increased or unchanged 

numbers of Foxp3- effector T cells (Teffs) along with manipulating 

the ΔNp63-miR181a-Sirt1 pathway. These findings provide 

novel mechanistic insights into how HCV induces T cell exhaustion 

via ΔNp63-miR-181a-DUSP6 signaling and premature T 

cell aging via ΔNp63-miR-181a-SIRT1 pathway. These two 

major pathways reveal new targets for therapeutic rejuvenation 

of impaired T cell responses during chronic viral infection. 

Disclosures: 

The following authors have nothing to disclose: Yun Zhou, Guangyu Li, Junping 

Ren, Ying Zhang, Jianqi Lian, Changxing Huang, Shunbin Ning, Jonathan P. 

Moorman, Zhansheng Jia, Zhi Q. Yao 

985 

Effects of Resistance Mutations of NS5A Inhibitor 

on Viral Production and Susceptibility to Anti-HCV 

Reagents in Recombinant Hepatitis C Viruses with NS5A 

of Genotype 1b 

Sayuri Nitta 1,2 , Yasuhiro Asahina 1 , Takaji Wakita 2 , Takanobu 

Kato 2 ; 1 Gastroenterology and Hepatology, Tokyo Medical and 

Dental University, Tokyo, Japan; 2 Virology II, National Institute of 

Infectious Diseases, Tokyo, Japan 

Backgrounds and Aims: The direct acting antiviral agents 

(DAAs) for hepatitis C virus (HCV) have strong inhibitory 

potency to HCV. However, using DAAs could cause the emergence 

of resistance mutations because these agents directly target 

HCV proteins. Amino acid substitutions at L31 and Y93 of 

non-structural protein 5A (NS5A) in HCV genotype 1b strains 

have been reported to confer resistance to NS5A inhibitor, 

daclatasvir (DCV). Furthermore, strains with these mutations 

are often detected in DCV treatment naïve patients. In this 

study, we assessed the effects of resistance mutations of DCV 

on HCV life cycle and susceptibility to DCV and other anti-HCV 

reagents by use of recombinant HCV harboring NS5A of genotype 

1b strain, Con1. Materials and Methods: We constructed 

recombinant HCV harboring NS5A of Con1 (JFH1-5ACon1), 

and introduced reported DCV resistance mutations, L31M, 

L31V, L31I and Y93H, in solely or in combination. In vitro transcribed 

full-length HCV RNA of these strains were transfected 

into Huh-7.5.1 cells and evaluated HCV replication and infections 

virus production by measuring the extra- and intra-cellular 

HCV core antigen (Ag) and infectivity titers. Susceptibilities of 

these strains to various anti-HCV reagents were also investigated. 

Results: All these strains are capable of replication in 

Huh-7.5.1 cells. At day 3 post-transfection, intra-cellular HCV 

core Ag levels were comparable in these strains transfected 

cells. However, extra-cellular HCV core Ag levels of strains with 

Y93H were about 2-fold higher than that of JFH-1/5ACon1. 

To assess the affecting step of this mutation in HCV life cycle, 

we determined the infectivity titers of these recombinant HCV. 

The strains with Y93H indicated higher infectivity titers than 

that of JFH-1/5ACon1. In the analysis of susceptibility to DCV, 

strains with mutation at L31 or Y93 showed the relatively mild 

or moderate resistance, while those with mutations at both L31 

and Y93 showed severe resistance, consistent with previous 

reports. Strains with DCV resistance mutations exhibited similar 

level susceptibility to IFNα, IFNλ1, IFNλ3 or RBV. Interestingly, 

the strains with Y93H mutation were more sensitive to protease 

inhibitor simeprevir (SMV) in comparison with JFH-1/5ACon1. 

Conclusions:In the in vitro study with recombinant HCV with 

NS5A of Con1, we indicated that the strains with Y93H 

enhanced infectious virus production. This observation suggests 

that once strains with Y93H emerged, it might be persistent 

after treatment. From our findings in susceptibility analysis to 

SMV, regimen including SMV could be considerable to treat 

the patients infected HCV with Y93H mutation. 

Disclosures: 

The following authors have nothing to disclose: Sayuri Nitta, Yasuhiro Asahina, 

Takaji Wakita, Takanobu Kato 

986 

The impact of HLA-DQs and IFNL4 variant on hepatitis C 

virus-induced liver inflammation 

Daiki Miki 1,2 , Hidenori Ochi 1,2 , C. Nelson Hayes 1,2 , Hiromi Abe 1,2 , 

Sakura Akamatsu 1,2 , Atsushi Ono 1,2 , Takashi Nakahara 1,2 , Yizhou 

Zhang 1,2 , Keiichi Masaki 1,2 , Hatsue Fujino 1,2 , Eisuke Miyaki 1,2 , 

Hiromi Kan 1,2 , Takuro Uchida 1,2 , Nobuhiko Hiraga 1,2 , Masataka 

Tsuge 1,2 , Tomokazu Kawaoka 1,2 , Michio Imamura 1,2 , Yoshiiku 

Kawakami 1,2 , Hiroshi Aikata 1,2 , Kazuaki Chayama 1,2 ; 1 Laboratory 

for Digestive Diseases, RIKEN Center for Integrative Medical 

Sciences, Hiroshima, Japan; 2 Department of Gastroenterology and 

Metabolism, Hiroshima University Hospital, Hiroshima, Japan 

Background & Aims: Several previous studies including genomewide 

association studies (GWAS) revealed that genetic variation 

in the IFNL3 (IL28B)-IFNL4 locus and the MHC region were 

associated not only with hepatitis C virus (HCV) persistence 

but also with HCV-induced cirrhosis and hepatocellular carcinoma. 

Because these variants appear to be associated with 

the progression of chronic HCV infection, they might play an


important role in chronic liver inflammation, although the role 

of these variants is still controversial. We performed large scale 

genotyping of IFNL4 and HLA-DQA1/B1, which have been 

reported as susceptibility loci for HCV persistence in European 

and Japanese populations, and analyzed the association 

between these genetic variants and the degree of histological 

inflammation in liver biopsies of patients with HCV. Methods: 

We directly sequenced exon 2 of HLA-DQA1 and HLA-DBB1 

using 775 Japanese patients with chronic HCV infection who 

underwent liver biopsies. In addition, we also investigated an 

IFNL4 variant (ss469415590) by multiplex-PCR-based Invader 

assay. We compared the frequencies of these genetic variants 

between patients with severe inflammation (necroinflammatory 

activity grade 2-3; n=514) and those with mild inflammation 

(activity grade 1; n=261). Results: The IFNL4 major homozygote 

TT/TT was more likely to be present in patients with 

severe activity grade (P = 0.034). Among common HLA-DQ 

alleles, HLA-DQA1*0101 and HLA-DQB1*0501 were 

observed more frequently (P < 0.05) in patients with mild activity 

(odds ratio [OR] = 0.58 and 0.57, respectively), whereas 

HLA-DQA1*0301 and HLA-DQB1*0401 were observed more 

frequently in those with severe activity (OR = 1.27 and 1.59, 

respectively). After adjustment for multiple testing, only HLA- 

DQA1*0101 was significantly associated with activity grade, 

and the effect size of this allele was not reduced by adjusting 

for IFNL4 variant and age at biopsy. Subsequent haplotype 

analysis revealed one significant protective haplotype, HLA- 

DQA1*0101-HLA-DQB1*0501 (P = 2.18 × 10 -4 , OR = 0.51), 

as well as one suggestive risk haplotype, HLA-DQA1*0301- 

HLA-DQB1*0401 (P = 5.12 × 10 -3 , OR = 1.48). Finally, results 

of multiple logistic regression analysis suggested that each 

haplotype and IFNL4 variant were independently associated 

with activity grade. Conclusions: We identified liver inflammation-associated 

HLA-DQ alleles and haplotypes that differed 

from the HCV persistence/clearance-associated allele, HLA- 

DQB1*03. Our findings suggest that HLA-DQ molecules might 

play different roles in liver inflammation and viral clearance, 

even as part of the same anti-viral immune response. 

Disclosures: 








The following authors have nothing to disclose: Daiki Miki, Hidenori Ochi, C. 

Nelson Hayes, Hiromi Abe, Sakura Akamatsu, Atsushi Ono, Takashi Nakahara, 

Yizhou Zhang, Keiichi Masaki, Hatsue Fujino, Eisuke Miyaki, Hiromi Kan, Takuro 

Uchida, Nobuhiko Hiraga, Masataka Tsuge, Tomokazu Kawaoka, Michio 

Imamura, Yoshiiku Kawakami, Hiroshi Aikata 

987 

The Hepatitis C Virus modulates its replication and 

the degradation of its RNA through the subversion of 

KHSRP function 

Patrice Bruscella 1 , Camille Baudesson 1 , Hassan Danso 1 , Michele 

Trabucchi 2 , Jean-Michel Pawlotsky 1 , Cyrille Feray 1 ; 1 Unit 955, 

INSERM, Creteil, France; 2 INSERM U 1065, C3M, Nice, France 

During infection, the hepatitis C virus (HCV) genome is exposed 

to degradation by the cellular RNA decay machinery. Indeed, 

HCV RNA is a target for cellular RNAses due to the lack of 

a 5’ methylated cap and of a 3’ poly-A tail and to the presence 

of a destabilizing RNA element (the 3’ poly-U region). 

MiR-122 has been shown to protect the HCV genome from 

5’-end degradation. The goal of this work was to unravel the 

role of KHSRP, an AU-Rich Element (ARE) mRNA-binding protein 

capable to recruit 5’- and 3’-RNAses such as XRN1 and 

DIS3, respectively, involved in miRNA maturation during HCV 

infection. Methods: Transient gene knockdown was performed 

by means of siRNA transfection. Viral RNA stability was evaluated 

by Northern-Blot analysis and qRT-PCR. Viral replication 

was measured using a modified JFH1 strain expressing 

Gaussia luciferase. Gene, pri-miRNA and miRNA expressions 

were quantified by qRT-PCR. The post-translational modifications 

and subcellular localization of endogenous KHSRP were 

studied by means of in situ Proximity Ligation Assay. Direct 

interaction of KHSRP with pri-miR-122, HCV RNA and ARE 

mRNA was studied by RNA Immunoprecipitation experiments 

using plasmids expressing KHSRP-HA, KHSRP-HA-S193D 

(phosphomimetic) or KHSRP-HA-S193A. Results: XRN1 and 

DIS3 silencing increased JFH1 replication in Huh7 hepatoma 

cells and the stability of nonreplicative viral RNA, whereas 

KHSRP silencing inhibited both viral replication and degradation 

of non-replicative HCV RNA. KHSRP silencing dramatically 

decreased miR-122 maturation. Using the JFH1 strain and a 

con1 subgenomic replicon, we showed that the expression of 

structural HCV proteins was associated with a decrease of the 

amount of KHSRP mRNA. During infection, the AKT-dependent 

phosphorylation of KHSRP at position Ser193 led to nuclear 

relocalization of phospho-KHSRP concomitantly to maturation 

of nuclear pri-miR-122. Proximity Ligation Assay showed co-localization 

of KHSRP, HCV NS5A protein and HCV RNA in 

the cytosol. Conclusions: HCV down-regulates KHSRP gene 

expression in hepatocytes, thus protecting HCV RNA from 

intracellular degradation by XRN1 and DIS3 RNAses. HCV 

infection also induces the phosphorylation and relocalization 

of phospho-KHSRP to the nucleus, where it induces miR-122 

maturation, thereby increasing HCV RNA stability and favoring 

replication. This so far unknown regulatory mechanism simultaneously 

promotes viral replication through miR-122 maturation 

and limits viral RNA degradation. 

Disclosures: 




The following authors have nothing to disclose: Patrice Bruscella, Camille Baudesson, 

Hassan Danso, Michele Trabucchi, Cyrille Feray 

988 

Liver pDCs from HCV-Infected Patients Are Primed for 

Interferon-alpha Production 

Erin H. Doyle 1 , Adeeb Rahman 2 , Arielle L. Klepper 1 , Sang Kim 6 , 

Brandy M. Haydel 3 , Sander S. Florman 4 , M. Isabel Fiel 5 , Thomas 

D. Schiano 4 , Andrea D. Branch 1 ; 1 Department of Liver Diseases, 

Icahn School of Medicine at Mount Sinai, New York, NY; 

2 Human Immune Monitoring Core, Icahn School of Medicine at 

Mount Sinai, New York, NY; 3 Center for Translational Transplant 

Research, Icahn School of Medicine at Mount Sinai, New York, 

NY; 4 Recanati Miller Transplantation Institute, The Mount Sinai 

Hospital, New York, NY; 5 Department of Pathology, The Mount 

Sinai Hospital, New York, NY; 6 Department of Anesthesiology, 

The Mount Sinai Hospital, New York, NY 

Background: Hepatitis C virus (HCV)-infected cells must be 

cleared from the liver to achieve a cure—whether through 

direct-acting antiviral drugs, or immune mechanisms. Interferon 

(IFN)α is an important anti-viral cytokine that is produced by 

plasmacytoid dendritic cells (pDCs), but its role in HCV clearance 

has been questioned because intrahepatic levels of IFNα 

mRNA are often below the limit of detection. This study combined 

a powerful new technique for cell phenotyping—mass 

cytometry time-of-flight (CyTOF)—with other methods to determine 

whether intrahepatic pDCs can and do produce IFNα.


Methods: Cells were isolated from explants of 20 HCV-infected 

patients undergoing liver transplantation. Blood was obtained 

prior to surgery. Liver and peripheral blood mononuclear cells 

(PBMCs) were analyzed by CyTOF, FACS, microarray, and 

multiplex cytokine assays with and without 4 hr stimulation with 

R848 (TLR7/8 agonist/single-stranded RNA mimic) or Poly 

I:C (TLR9 agonist/double-stranded RNA analogue). Results: 

CyTOF delineated monocytes/macrophages, DC, NK, T and 

B cell subsets and revealed that pDCs are the only liver or 

blood cell that produces IFNα. Cytokine assays showed that 

liver pDCs were primed for robust IFNα production: secreted 

levels increased from 0.8 to 344.7 pg/mL after 4 hrs of R848 

stimulation. This increase was much higher than that of blood 

pDCs, which increased production from 2.4 to 115 pg/mL, 

p


as shown by increased ISG induction. Interestingly, SeP was 

found to regulate IFN signal transduction independent of the 

JAK-STAT pathway. In vivo, the ISGs Mx-1 and Oas2 were 

significantly increased in SepKO mice compared with WT mice 

following PolyI:C injection. To explore the clinical significance, 

hepatic gene expression was analyzed in 66 CHC patients; 

SeP was upregulated in the early stage of liver fibrosis and was 

significantly correlated with a poor outcome of IFN therapy. 

Conclusion: In CHC, SeP impaired innate immunity. The present 

findings would be useful for the development of novel therapeutic 

strategies for patients with chronic liver disease with various 

etiologies including NASH and CHC. 

Disclosures: 

Stanley M. Lemon - Advisory Committees or Review Panels: Merck, Santaris, 

Abbott, Gilead; Consulting: Achillion, Idenix; Grant/Research Support: Merck, 

Tibotec, Scynexis; Speaking and Teaching: Hoffman LaRoche 






The following authors have nothing to disclose: Kazuhisa Murai, Masao Honda, 

Tetsuro Shimakami, Takayoshi Shirasaki, Hirofumi Misu, Toshinari Takamura, 

Seishi Murakami 

991 

Long noncoding RNAs regulate HCV replication and 

ISGs in a Jak-STAT independent manner 

Xiao Liu, Jian Hong, Dachuan Cai, Sae Hwan Lee, Dahlene Fusco, 

Esperance A. Schaefer, Cynthia Brisac, Anna Lidofsky, Wenyu Lin, 

Raymond T. Chung; Gastrointestinal Unit, Massachusetts General 

Hospital, Harvard Medical School, Boston, MA 

Background/ Aims: Long non-coding RNAs (lncRNAs) involve 

in many important cellular process regulations. However, their 

roles in the innate immune response including type-1 interferon 

(IFN) response to HCV infection are not well understood. 

We previously identified SART1, a host protein involved in 

RNA splicing and pre-mRNA processing, as a regulator of 

IFN antiviral effects (Lin et al, J Hepatology 2015, 62:1024). 

We sought to explore lncRNA regulators of HCV replication, 

the JAK-STAT pathway, and interferon-stimulated genes (ISGs). 

Methods: We performed SART1 siRNA knockdown and RNA- 

Seq in Huh7.5.1 cells with or without IFN treatment. Bioinformatic 

Homo sapiens Genome Browser Gateway analysis was 

performed to predict the location of lncRNAs and adjacent 

coding genes. Selected lncRNAs, Jak-STAT pathway genes and 

ISGs and their proteins, together with HCV replication, were 

monitored by qRT-PCR and Western blot in the JFH1 HCV infectious 

model. Results: We obtained 6699 lncRNAs regulated 

by SART1 and IFN in RNA-Seq. Our bioinformatic analysis 

of RNA-Seq data identified a cluster of 60 lncRNAs involved 

in IFN’s antiviral regulation. qRT-PCR confirmed that expression 

of 14 of these lncRNAs was consistent with the RNA-Seq 

data. We then identified 4 anti-sense lncRNA related to ISGs 

which negatively regulated the adjacent coding gene expression 

based on previous reports. Of these, we selected lncRNA 

RP11-670E13.5 (lncRNA-TRIM25) for further characterization 

of its regulation of HCV. We found that knockdown of lncRNA 

RP11-670E13.5 significantly increased HCV replication compared 

to Neg siRNA. We also found that siRNA lncRNA RP11- 

670E13.5 significantly reduced TRIM25, MX1, ISG15 mRNA 

compared to Neg siRNA in JFH1-infected Huh7.5.1 cells. 

Interestingly, STAT1, JAK1, OAS3, and RIG-I mRNA expression 

were not affected by the knock-down of lncRNA RP11- 

670E13.5. Moreover, we found that siRNA to STAT1 had no 

effect on lncRNA RP11-670E13.5 expression. These data suggest 

that lncRNA RP11-670E13.5 regulation of HCV is independent 

of the Jak-STAT signaling pathway. Conclusion: We 

found that lncRNA RP11-670E13.5 has important regulatory 

functions of antiviral innate immunity in the JFH1 HCV model. 

Our data suggested that lncRNA RP11-670E13.5 regulates 

HCV replication independently of the JAK-STAT pathway. We 

speculate that RP11-670E13.5 exerts its regulatory function on 

the enhancer and promoter activation of nearby protein-coding 

(target) genes such as TRIM25, MX1, and ISG15. Further characterization 

of the mechanism by which selected lncRNAs that 

associate with these genes regulate HCV replication and ISGs 

are warranted. 

Disclosures: 

Dahlene Fusco - Grant/Research Support: Gilead 


Merck, BMS 

The following authors have nothing to disclose: Xiao Liu, Jian Hong, Dachuan 

Cai, Sae Hwan Lee, Esperance A. Schaefer, Cynthia Brisac, Anna Lidofsky, 

Wenyu Lin 

992 

Evaluation of protective and therapeutic strategies 

against hepatitis C virus in a syngenic transplantation 

mouse model 

Matti Sällberg 1 , Sepideh Levander 1 , Fredrik Holmstom 1 , Gustaf 

Ahlén 1 , Lars Frelin 1 , Gang Long 2 , Daniel Rupp 2 , Ralf Bartenschlager 

2 ; 1 Laboratory Medicine, Karolinska Institutet, Stockholm, 

Sweden; 2 Molecular Virology and Infectious Diseases, University 

of Heidelberg, Heidelberg, Germany 

Key features of the hepatitis C virus (HCV) are its high genetic 

plasticity and its ability to establish chronic infection in hepatocytes. 

Development of prophylactic vaccines is hampered by 

the lack of immune competent small animal models supporting 

robust HCV replication. Herein, we devised an immune competent 

mouse model that is based on the syngenic transplantation 

of H-2 b -restricted Hep56 cells containing a self-replicating 

subgenomic HCV replicon RNA of genotype (gt) 2a. To allow 

non-invasive in vivo monitoring of tumor growth, cells were 

also stably transfected with a luciferase (Luc) reporter gene. 

As controls, Hep56 cells stably expressing an enzymatically 

active HCV NS3/4A (gt2a) protease complex were used. 

Upon subcutaneous injection, all cell lines generated palpable 

tumors with sizes peaking around day 8-16 post inoculation. 

Tumor growth correlated with an accumulation of inflammatory 

cells and central necrotic areas, concomitant with a decrease 

of HCV RNA and Luc DNA copy numbers. DNA vaccination 

resulted in a robust NS3/4A(gt2a)-specific CD4+ and CD8+ 

T cell response protecting wild type, but not HCV NS3/4A(gt1a)-transgenic 

(Tg) mice against HCV replicon cell tumors suggesting 

cross-genotypic tolerance in NS3/4A(gt1a)-Tg mice. 

Hep56 cells stably expressing gt2a NS3/4A primed and 

boosted a strong CTL-dominated response, whereas the HCV 

replicon cells did not, despite comparable levels of antigen 

expression. Thus, similar to human infection, hepatocytes with 

replicating HCV RNA appear to be poorly immunogenic arguing 

that the replicon-cell based syngenic transplantation model 

shares features of infected hepatocytes in patients. Unlike other 

models, syngenic transplantation of HCV replicon cells can 

be rapidly applied to genetically modified immune competent 

mice of the widely used C57BL/6J background for the study 

of HCV-specific cellular immune responses. This model will be 

highly useful in vaccine development for hepatitis C 

Disclosures: 

Matti Sällberg - Consulting: Chrontech Pharma AB, Abbvie 

The following authors have nothing to disclose: Sepideh Levander, Fredrik Holmstom, 

Gustaf Ahlén, Lars Frelin, Gang Long, Daniel Rupp, Ralf Bartenschlager


993 

Analysis of AL-516, a Novel Potent Nucleotide Analog, 

Combination Drug Interactions in the Hepatitis C Virus 

(HCV) Subgenomic Replicon System 

Hua Tan, Kenneth Shaw, vivek Rajwanshi, Gary Wang, Leo Beigelman, 

David B. Smith, Lawrence M. Blatt, Julian A. Symons; 

Alios, South San Francisco, CA 

Background: Nucleotide analogs have emerged as an important 

component of interferon (IFN)-free combination therapies 

for the treatment of chronic hepatitis C (CHC) based on their 

potent activity and high barrier to the generation of viral resistance. 

AL-516, a novel monophosphate prodrug of a guanosine-based 

nucleotide analog, has been identified as a potent 

and selective inhibitor of NS5B-directed HCV RNA replication 

in the cell based replicon system. In this study, inhibition of the 

HCV replicon by AL-516 was examined in pairwise combinations 

with multiple compounds either registered for the treatment 

of CHC or currently in clinical development. Methods: Studies 

were performed using a Huh-7 cell line expressing a Firefly 

luciferase-encoding HCV 1b subgenomic replicon. Compounds 

were added to cells in a checkerboard fashion and inhibition 

of HCV replication measured by luminescence. Data were analyzed 

using two drug interaction models; Isobologram analysis 

using the Loewe additivity model and the Bliss-Independence 

model using Pritchard’s MacSynergy II software. Results: In 

the HCV 1b replicon, AL-516 exhibits potent antiviral activity 

with an EC 50 

of 6.5 nM. When tested in pairwise combinations 

with representative members of the NS3/4A protease 

inhibitors, NS5A inhibitors, nucleoside/tide and non-nucleoside 

polymerase inhibitors, cyclophilin A inhibitors, type I and 

type III IFN’s or RBV, AL-516 demonstrated significant synergy 

(>100 mM2%), synergy (25-100 mM2%) or additivity (0-25 

mM2%), no antagonistic effects were observed. For approved 

compounds, combination of AL-516 with the NS3/4A protease 

inhibitor, simeprevir, exhibited significant synergy with 

a synergy volume of 114.1 mM2%. AL-516 also exhibited 

significant synergistic interactions with the HCV NS5A inhibitor, 

daclatasvir, (117.1 mM2%) and additive-to-synergistic 

interactions with the investigational uridine-based nucleoside 

polymerase inhibitor, AL-335, (33.5 mM2%). Conclusions: 

Future IFN-free therapy for CHC will require a combination 

of compounds with different mechanisms of action. AL-516 

demonstrates an in vitro antiviral profile that suggests it may 

become an important component of IFN-free combination therapy. 

To this end, AL-516 is currently advancing towards human 

clinical trials for CHC. 

Disclosures: 

Kenneth Shaw - Employment: Alios Biopharma 

David B. Smith - Employment: Alios BioPharma 

Lawrence M. Blatt - Management Position: Alios BioPharma 

Julian A. Symons - Employment: Alios BioPharma 

The following authors have nothing to disclose: Hua Tan, vivek Rajwanshi, Gary 

Wang, Leo Beigelman 

994 

Regulation of Hepatitis C Virus Infection by Long 

Non-Coding RNAs 

Tetsuro Shimakami, Masao Honda, Takayoshi Shirasaki, Fanwei 

Liu, Masaya Funaki, Kazuhisa Murai, Shuichi Kaneko; Kanazawa 

University, Kanazawa, Japan 

Background Recently, large numbers of non-coding RNAs 

have been identified that reportedly have a wide range of 

functions. Non-coding RNAs are categorized into two groups: 

the first is composed of short non-coding RNAs, such as siRNA 

and miRNA, and the second contains long non-coding RNAs, 

which are generally more than 200 ntds in length and possess 

a 5ʹ cap and 3ʹ poly-A tail. The role of short non-coding RNAs 

in hepatitis C virus (HCV) infection, such as miR-122, has been 

characterized considerably more than that of long non-coding 

RNAs. Here, we studied the role of long non-coding RNAs 

in HCV infection. Method To identify long non-coding RNAs 

whose expression is specifically altered by HCV infection, 

Huh-7.5 cells were infected with the HJ3-5 virus, which is a 

Ia/IIa chimera. Total cellular RNAs were extracted, followed 

by amplification of only poly-A tail RNAs, and subjected to 

sequence analysis using a next-generation sequencer. We 

selected 26 candidate long non-coding RNAs whose expression 

was specifically and significantly increased by HCV infection. 

Next, we designed 2–3 siRNAs for each candidate and 

tested the effect of knockdown of each long non-coding RNA in 

HCV cell culture. Result The knockdown of 4 long non-coding 

RNAs significantly suppressed HCV replication. From these, 

we focused on lncRNA-H, which has already been registered 

in the lncRNA database and whose expression in human livers 

has been confirmed by several reports; however, its role in 

HCV infection remains unknown. The expression of lncRNA-H 

was induced by HCV infection in a multiplicity of infection-dependent 

manner in HCV cell culture, and NS5A seemed to be 

the inducer of lncRNA-H expression. This induction was also 

observed in an HCV-infected chimpanzee as well as HCV-infected 

chimeric mice. Furthermore, when we compared the 

expressions of lncRNA-H in human liver between pre- and 

post-eradication of HCV, the expression of lncRNA-H was significantly 

decreased by the eradication of HCV. The knockdown 

of lncRNA-H by siRNAs suppressed HCV replication 

as well as infectious virus production, in accordance with the 

knockdown of lncRNA-H. This effect seemed to be through the 

suppression of HCV-IRES-dependent translation. Furthermore, 

we compared the amount of lncRNA-H in HCV-infected human 

livers and serums derived from IL28B genotype (rs8099917) 

major and minor allele patients. Interestingly, the amount of 

lncRNA-H from minor allele patients was significantly higher 

than in patients with the major both in livers and serums. Discussion 

These results suggest that lncRNA-H could be a new 

target for the treatment of HCV infection as well as a new 

biomarker for interferon-based therapies. 

Disclosures: 






The following authors have nothing to disclose: Tetsuro Shimakami, Masao 

Honda, Takayoshi Shirasaki, Fanwei Liu, Masaya Funaki, Kazuhisa Murai 

995 

Iron chelation restores mitophagy suppressed by hepatitis 

C virus 

Yuichi Hara 1 , Sohji Nishina 1 , Tasuku Hirayama 2 , Hideko Nagasawa 

2 , Keisuke Hino 1 ; 1 Kawasaki Medical College, Kurashiki, 

Japan; 2 Gifu Pharmaceutical University, Gifu, Japan 

Background and aim: Oxidative stress is present in chronic 

hepatitis C to a greater degree than in other inflammatory 

liver disease and closely related to disease progression. We 

recently reported that hepatitis C virus (HCV) core protein 

suppresses mitophagy by interacting with Parkin (Am J Pathol 

2014), which may amplify and sustain HCV-induced oxidative 

stress. Therefore, the restoration of mitophagy is a critical therapeutic 

intervention for preventing disease progression including 

liver cancer development in chronic hepatitis C. On the other


hand, iron loss has been reported to trigger PINK1/Parkin-independent 

mitophagy. The aim of this study was to determine 

whether iron chelation restores mitophagy suppressed by HCV. 

Methods: HCV-JFH1 infected cells were treated for 24h with a 

final concentration of 1mM deferiprone (DFP), iron chelator. 

Transgenic mice expressing the HCV polyprotein were administered 

0.075g of DFP dissolved in water through gastric tube 

for 2 months. The effect of DFP on mitophagy and mitochondrial 

function was examined in vitro and in vivo. Results: As 

reported previously, suppressed mitophagy was confirmed in 

HCV-JFH1 infected cells. DFP treatment increased the expression 

of microtubule-associated protein light chain 3 (LC3)-II and 

decreased the expression of p62 in dose-dependent manner, 

but did not affect translocation of Parkin to mitochondria in 

HCV-JFH1 infected cells. Electron microscopy also revealed 

that DFP significantly increased the number of mitophagosomes 

in HCV-JFH1 infected cells and liver specimens from transgenic 

mice expressing the HCV polyprotein. These results showed 

that DFP restored mitophagy suppressed by HCV core protein. 

DFP also decreased mitochondrial membrane potential 

and reactive oxygen species (ROS) production, but not ATP 

production in HCV-JFH1 infected cells. Next, we measured 

mitochondrial ferrous iron content, using a highly specific 

turn-on fluorescent probe (Ac-MT-FluNox1) that is specific to 

labile ferrous iron in mitochondria. Mitochondrial ferrous iron 

content was significantly higher in cells with HCV-JFH1 infection 

than in those without HCV-JFH1 infection in the absence of 

DFP treatment. DFP decreased mitochondrial ferrous iron content 

in dose dependent manner regardless of HCV infection. 

Conclusion: These results indicated that iron chelation restores 

mitophagy suppressed by HCV. The mechanisms underlying 

mitophagy induced by iron chelation remains to be unknown, 

but decrease in mitochondrial ferrous iron and disrupted mitochondrial 

function may be critical for inducing mitophagy. 

Disclosures: 

The following authors have nothing to disclose: Yuichi Hara, Sohji Nishina, 

Tasuku Hirayama, Hideko Nagasawa, Keisuke Hino 

996 

Cell-specific peripheral innate immune responses to 

immunomodulatory stimulation in chronic HCV 

Jacinta A. Holmes 1,2 , Narelle A. Skinner 2 , Mario Congiu 2 , Rosemary 

M. Millen 2 , Lijia Yu 2 , William Sievert 3 , Paul V. Desmond 1 , 

Kumar Visvanathan 2 , Alex J. Thompson 1,2 ; 1 Gastroenterology, St 

Vincent’s Hospital, Fitzroy, VIC, Australia; 2 Immunology Research 

Centre, St Vincent’s Hospital, Melbourne, VIC, Australia; 3 Gastroenterology, 

Monash Health, Monash University, Melbourne, VIC, 

Australia 

INTRODUCTION: Our preliminary data showed chronic HCV 

(CHC) patients (pts) who achieve SVR have potent and early 

immune responses, with chemokine, cytokine and interferon-stimulated 

gene (ISG) induction. The pattern is similar in CC 

IFNL3 genotype (gt) pts. There are no data regarding which 

cell types contribute most to immunomodulatory responses or 

which immune pathways are important in CHC. We hypothesised 

monocytes (MC) would display the greatest induction 

of ISGs post-stimulation, particularly in CC pts, and viral sensing 

TLR pathways would generate the greatest responses. We 

performed a pilot study to characterise cell-specific ISGs in 

CHC PBMCs pre/post immunomodulatory stimulation. METH- 

ODS: Treatment-naïve HCV1 F0-3 pts were enrolled. PBMCs 

were stimulated with TLR2, 3, 4, 7/8, 9 ligands and IFNa. 

PBMCs were sorted into NKs, NKTs and MC using a cell 

sorter. RNA was extracted, and ISG expression (MX1/ISG15) 

measured (rtPCR). Levels were compared between cell types, 

pre/post-stimulation (fold-change), and according to IFNL3 gt. 

RESULTS: 10 pts were included: 50% CC, 40% male, median 

age 53 years. At baseline non-CC pts demonstrated higher ISG 

mRNA, predominantly from NK/NKT cells (Fig.1a). Post-stimulation, 

ISG induction was significantly higher in MC (131 to 

168-fold increase) than NK/NKT cells (maximum 60-fold-increase) 

following TLR7/8 and IFNa stimulation (Fig.1b). MC 

from CC pts demonstrated a 2-3 fold-increase in MX1 than 

non-CC pts following stimulation (Fig.1c). CONCLUSIONS: 

The data identify MC as key immunomodulatory cells, and 

demonstrate the importance of the TLR7/8 pathway in immune 

responses in CHC, supporting clinical development of TLR7 

agonists. Furthermore, MC from CC IFNL3 gt pts demonstrated 

greater immune reactivity, and may contribute to the more 

potent immune responses observed during therapy, leading to 

greater viral clearance. Unraveling these differences in immune 

responses may lead to new therapeutic targets, particularly for 

patients who fail IFN-free therapies. 

Disclosures: 

Jacinta A. Holmes - Grant/Research Support: Roche, Merck, AASLD, St Vincent’s 

Hospital Research Endowment Fund 

William Sievert - Advisory Committees or Review Panels: Gilead Sciences, Bristol 

Myers Squibb, Merck; Speaking and Teaching: Merck, Bristol Myers Squibb 




Abbvie, BMS 

The following authors have nothing to disclose: Narelle A. Skinner, Mario Congiu, 

Rosemary M. Millen, Lijia Yu, Paul V. Desmond, Kumar Visvanathan 

997 

Role of very-low-density lipoprotein (VLDL) biogenesis 

in hepatitis C virus (HCV) production: a reassessment in 

primary human adult hepatocytes 

Veronique Pene 1 , Maxime Villaret 1 , Matthieu Lemasson 1 , Lynda 

Aoudjehane 2,3 , Jean-François Méritet 1,4 , Filomena Conti 3,5 , Yvon 

Calmus 3,5 , Arielle R. Rosenberg 1,4 ; 1 EA 4474 “Hepatitis C Virology”, 

Université Paris Descartes, Paris, France; 2 Human HepCell, 

Hôpital Saint-Antoine, Paris, France; 3 UMRS 938 “CDR Saint-Antoine”, 

Inserm, Paris, France; 4 Service de Virologie, AP-HP, Hôpital 

Cochin, Paris, France; 5 Unité de Transplantation Hépatique, 

AP-HP, Hôpital Pitié-Salpêtrière, Paris, France 

Background and Aim. In the blood of HCV-infected patients, 

infectivity is mainly supported by viral particles associated 

with triglyceride-rich lipoproteins containing apolipoprotein B 

(ApoB) and ApoE. These complexes are believed to assemble 

within the hepatocyte, which is both the primary replication site


of HCV and the cell type specialized in the secretion of VLDL. 

The known requirements for VLDL biogenesis are (i) ApoB, a 

structural component essential for VLDL scaffolding, and (ii) 

microsomal triglyceride transfer protein (MTP), the rate-limiting 

enzyme that lipidates ApoB. However, studies with the classical 

HCV culture system in hepatocarcinoma-derived Huh-7 sublines 

suggested that MTP inhibitors might not efficiently block 

HCV production unless high, cytotoxic concentrations are used. 

Moreover, the role of ApoB remains a matter of controversy 

whereas ApoE appears to be a prerequisite for HCV production 

in cell line culture. Here we have reassessed the role of VLDL 

biogenesis in HCV morphogenesis and secretion using a more 

relevant HCV culture system in primary human adult hepatocytes 

(PHH), which, contrary to Huh-7 cells, secrete authentic 

VLDL and infectious particles. Methods. PHH were infected with 

the HCV strain JFH1, and either treated with increasing doses 

of various MTP inhibitors or depleted for ApoB or ApoE (RNA 

interference technologies). Cultures were evaluated for production 

of intracellular and extracellular infectious virus (focus-formation 

assay), viral load (RT-qPCR), secretion of ApoB and 

ApoE (ELISA), and cytotoxic effects. Results. The pharmacological 

MTP inhibitor CP-346086, which efficiently inhibited 

VLDL secretion in PHH, also reduced both HCV production 

and infectivity at moderate, non-cytotoxic doses. The grapefruit 

flavonoid naringenin, reported to inhibit MTP activity indirectly 

through a PPARa-mediated mechanism, induced a significant 

reduction of HCV production even at doses that did not reduce 

VLDL secretion. ApoB depletion did not seem to affect HCV production 

or infectivity significantly even though it inhibited VLDL 

biogenesis. ApoE depletion caused only a moderate reduction 

of HCV infectivity in PHH whereas it almost completely abolished 

HCV production in Huh-7.5.1 cells. Conclusion. These 

data in differentiated human hepatocytes show that neither 

ApoB nor ApoE is essential for HCV morphogenesis whereas 

MTP function is absolutely required for HCV production and 

infectivity, albeit not necessarily via its role in VLDL biogenesis. 

MTP inhibitors, either direct (CP-346086) or indirect (naringenin), 

appear as promising host-targeting drugs to combat 

HCV infection in complement with directly acting antivirals. 

Disclosures: 

The following authors have nothing to disclose: Veronique Pene, Maxime Villaret, 

Matthieu Lemasson, Lynda Aoudjehane, Jean-François Méritet, Filomena Conti, 

Yvon Calmus, Arielle R. Rosenberg 

998 

A miR-122/miR-224-based model to identify severe 

fibrosis and cirrhosis in patients with chronic hepatitis C 

Kevin Appourchaux 2 , Emilie Estrabaud 2 , Matthieu Resche-Rigon 

3,1 , Martine Lapalus 2 , Michelle Martinot-Peignoux 2 , Nathalie 

Boyer 4 , Michel Vidaud 5 , Pierre Bedossa 6 , Patrick Marcellin 2 , Tarik 

Asselah 2,4 ; 1 INSERM, Paris, France; 2 INSERM UMR1149, Paris, 

France; 3 INSERM UMR1153, Paris, France; 4 Hepatology, Beaujon 

Hospital, Clichy, France; 5 INSERM UMR745, Paris, France; 6 Service 

d’Anatomie Pathologique, Beaujon Hospital, Clichy, France 

Background and aims Staging fibrosis is crucial in patients 

with chronic hepatitis C (CHC) because it reflects the prognosis. 

MiRNAs regulate the expression of up to 60% of mRNAs. 

The liver-enriched miR-122 enhances HCV replication. MiRNAs 

are increasingly investigated as biomarkers because of their 

high stability compared to mRNAs and proteins. We aimed 

to identify miRNAs associated with fibrosis in patients with 

CHC. Patients and Methods A total of 202 patients with CHC 

were consecutively enrolled. The inclusion criteria were having 

at least one biopsy and no HCC. Serums and biopsies 

samples were available for respectively 106 and 86 patients 

(26 paired liver/serum). Among patients with available serum, 

63.2% were male, the mean age was 48.8 years and the 

mean HCV viral load was 5.81 logUI/mL. According to the 

Metavir scoring system, 24.5%, 16%, 34.9%, 24.6% of the 

patients were respectively F1, F2, F3 and F4. Among patients 

with available biopsies, 54.2% were male, the mean age was 

49.9 years and the mean HCV viral load was 5,6 logUI/mL. 

Regarding the stages of fibrosis, 27,1%, 31,2%, 22,9%, and 

18,8% of the patients were respectively F1, F2, F3 and F4. 

MiR-20a, -27b, -29a, -92a, -122, -146a, -222, -224 were 

selected for the study because of their role during liver fibrosis. 

The expression of miRNAs was assessed by RT-qPCR. Results 

An increased expression of hepatic miR-224 (p=0.003) was 

observed in patients with mild and moderate fibrosis (F1-F2) 

compared to those with severe fibrosis and cirrhosis (F3-F4). 

A reduction of hepatic miR-122 (p=0.008) was observed in 

patients with F3-F4 compared to those with F1-F2. For hepatic 

miR-20a, -27b, -29a, -92a, -122 and -146a, no difference was 

observed between patients with F3-F4 and those with F1-F2. 

In the serums, none of the miRNAs tested, was significantly 

deregulated between F3-F4 and F1-F2. The multivariate analysis 

of clinical data (albumin, platelets count, aspartate aminotransferase 

(AST), alkaline phosphatase) and hepatic miRNAs, 

allowed us to build a model combining hepatic miR-122 and 

miR-224, platelets count and AST. The AUC of the model was 

0.90 while in the same cohort, FIB-4 and APRI had, respectively, 

an AUC of 0.78 and 0.85. Conclusions Interestingly, 

hepatic miR-122 and miR-224 were respectively decreased 

and increased in patients with severe fibrosis and cirrhosis. 

Increased expression of miR-224 might suggest a premalignant 

condition to HCC. The model combining the assessment of 

hepatic miR-122 and miR-224, platelets count and AST was 

more accurate than FIB-4 and APRI to distinguish patients with 

F3-F4 from those with F1-F2. 

Disclosures: 







Abbvie 



The following authors have nothing to disclose: Kevin Appourchaux, Emilie 

Estrabaud, Matthieu Resche-Rigon, Martine Lapalus, Michelle Martinot-Peignoux, 

Michel Vidaud, Pierre Bedossa 

999 

Different HCV genotypes have different genetic barriers 

to resistance to N5A inhibitors, explaining different 

treatment outcomes 

Slim FOURATI 1,2 , Stephane Chevaliez 1,2 , Alexandre Soulier 1,2 , 

Marion Lavert 1,2 , Lila Poiteau 1,2 , Jean-Michel Pawlotsky 1,2 ; 1 Virology, 

Hôpital Henri Mondor, AP-HP,, National Reference Center for 

Viral Hepatitis B, C and Delta,, Créteil, France; 2 INSERM U955, 

Université Paris-Est, Créteil, France 

The genetic barrier to resistance of an antiviral drug depends 

on the number and type of nucleotide substitutions required to 

overcome the drug’s selective pressure. It is a key factor for 

the development of HCV drug resistance in the clinical setting. 

The goal of this study was to assess whether different 

HCV genotypes and subtypes have different genetic barriers 

to resistance to NS5A inhibitors that could explain the clinical 

results of NS5A inhibitor-containing regimens. Methods: 

744 unselected sequences extracted from the European HCV 

database (https://euhcvdb.ibcp.fr) were examined at 9 amino


acid positions located in domain I of NS5A (residues 28, 29, 

30, 31, 32, 58, 62, 92 and 93); amino acid changes at these 

positions are known to be responsible for reduced susceptibility 

to one or several NS5A inhibitors, including ledipasvir, 

daclatasvir, ombitasvir and/or elbasvir. The genetic barrier 

was calculated as the sum of transitions (scored as 1) and/ 

or transversions (scored as 2.5) required for evolution to any 

drug resistance mutation in NS5A. Comparison between genotypes 

and subtypes (1a, 1b) was performed based on the most 

frequent codon in the genotype/subtype. Results: The genetic 

barrier was identical across all genotypes only at NS5A positions 

29 and 32 (P29S and P32L scored 1 for all 6 genotypes). 

In contrast, a higher barrier to resistance was found in subtype 

1b than in subtype 1a at positions M/L28T (score: 3.5 

vs 1, respectively), M/L28V (2.5 vs 1), M/L28A (3.5 vs 2), 

Q/R30E/H/K (3.5 vs 2.5) and H/P58D (7.5 vs 2.5). Conversely, 

subtype 1b displayed a lower genetic barrier than 1a 

to acquire H/P58S (1 vs 3.5, respectively). Genotypes 3 and 

4 displayed a lower genetic barrier to acquire A/R30S than 

genotype 1 (2.5 vs 3.5, respectively). Residue Y93 was highly 

conserved across genotypes 1, 2 and 3. Indeed, in contrast to 

genotype 6 which displayed a high genetic barrier to acquire 

93C (score 2.5) and 93H (score 5), these amino acid substitutions 

required a minimal score of 1 for genotypes 1, 2 and 3. 

Conclusions: HCV subtype 1a displays a lower genetic barrier 

to resistance than subtype 1b at several NS5A positions at the 

nucleotide level. This could account for the higher rate of failure 

of NS5A inhibitor-containing regimens in patients infected with 

HCV subtype 1a. In addition, genotype-specific amino acid 

changes (e.g. R30S in genotype 4, P58S in subtype 1b) could 

be explained by a differential genetic barrier between different 

genotypes and subtypes. 

Disclosures: 

Slim FOURATI - Speaking and Teaching: Gilead 

Stephane Chevaliez - Advisory Committees or Review Panels: Janssen; Speaking 

and Teaching: Gilead, BMS 




The following authors have nothing to disclose: Alexandre Soulier, Marion Lavert, 

Lila Poiteau 

1000 

Establishment and Characterization of a New Permissive 

Cell Line for HCV Infection 

Hitoshi Omura, Tetsuro Shimakami, Takayoshi Shirasaki, Kazuhisa 

Murai, Masaya Funaki, Fanwei Liu, Takehiro Hayashi, Taro 

Yamashita, Masao Honda, Shuichi Kaneko; Kanazawa university, 

Kanazawa city, Japan 

Background The in vivo HCV infection model has been needed 

to clarify the mechanism of persistent infection of HCV in human 

livers. Huh-7.5, which is a subline of human hepatoma cell line, 

Huh-7, has been used most frequently for propagation of HCV 

due to its vigorous ability to support for HCV replication. Huh- 

7.5 is quite useful for the development of many direct-acting 

antivirals (DAAs). Huh-7.5 is known to have the defect in its 

antiviral interferon signaling, therefore, a new cell line with 

intact interferon signaling is needed to mimic HCV infection 

in human livers. Here, we have established and characterized 

a new permissive cell line for HCV infection with intact innate 

antiviral interferon signaling. Method The human hepatoma 

tissue was obtained from an HCV-infected patient at a surgical 

resection, and then transplanted to mice. The adherent cells to 

mice were removed, followed by the MACS cell separation to 

remove mice fibroblasts, and used for developing a new cell 

line, designated as KH cell. We examined the permissiveness 

of KH cells for HCV and characterized by comparing with 

Huh-7.5 cells. Result The HCV RNA was no longer detected in 

KH cells by qRT-PCR from the original patient. The karyotype 

analysis for KH and Huh-7.5 cells revealed that the two cell 

lines were distinct from each other. KH cells could support HCV 

replication as efficiently as Huh-7.5 cells after transfection of in 

vitro transcribed HCV RNA. Additionally, the infection of cell 

culture-derived HCV to KH cells showed the entry of HCV into 

the cells, followed by efficient replication. KH cells replicating 

HCV could also produce infectious viruses, suggesting that KH 

cells had the ability to support a full life cycle of HCV as could 

Huh-7.5 cells. KH cells had same amount of miR-122 as did 

Huh-7.5. We measured EC50 for IFN-α in KH and Huh-7.5 

cells. We then compared EC50 of IFN-α between both and 

found a similarity. Interestingly, the sequence analysis of RIG-I 

at its 55 amino acid position for KH cells showed wild type, 

T55. RIG-I from Huh-7.5 cells was mutant, T55I, which is a 

representative mutation of Huh-7.5 cells. In accordance with 

this result about RIG-I, the transfection of HCV RNA into KH 

cells induced robust ISGs induction, such as MX1, IFN-β, and 

OAS2, on the other hand, no induction was observed in Huh- 

7.5 cells. Discussion We have established and characterized a 

new permissive cell line for HCV infection with intact antiviral 

interferon signaling. This cell line could be useful to understand 

the underlying mechanism of persistent HCV infection under 

pressure of host antiviral innate immunity. 

Disclosures: 






The following authors have nothing to disclose: Hitoshi Omura, Tetsuro Shimakami, 

Takayoshi Shirasaki, Kazuhisa Murai, Masaya Funaki, Fanwei Liu, Takehiro 

Hayashi, Taro Yamashita, Masao Honda 

1001 

PD-1 blockade enhances cytopathic control of HCV by 

antiviral CD8 T cells in infectious HCV co-culture model 

Keisuke Ojiro 1 , Xiaowang Qu 1,2 , Jang-June Park 1 , Hyosun Cho 1,3 , 

Kyong-Mi Chang 1 ; 1 Division of Gastroenterology, University of 

Pennsylvania, Philadelphia, PA; 2 Translational Medicine Institute, 

University of South China, Chenzhou, China; 3 Pharmacy, Duksung 

Women’s University, Seoul, Korea (the Republic of) 

Background: T-cell expression of inhibitory costimulatory 

receptor PD-1 and CTLA-4 in chronic viral infection and tumor 

environment is associated with functional tolerance that may 

be reversed by antibody-mediated blockade in vitro and in 

vivo. Thus, PD-1 blockade is in both pre-clinical and clinical 

development in various settings. In this study, we examined 

the effect of PD-1 blockade on T cell mediated virus control 

and hepatocyte survival using the infectious HCV co-culture 

system. Method: Target cells for HCV replication and antigen 

presentation were established by lentiviral transduction of HLA 

A2.1, PDL1 and/or GFP expression in Huh7.5 hepatoma 

cells. Highly infectious JFH derived strain Jc1/Gluc2A clone 

(genotype 2a) was modified by site-directed mutagenesis to 

encode well-defined genotype 1a-derived NS3 1073 or NS5B 

2594 epitopes. HCV-specific CD8 T cells were expanded from 

peripheral lymphocytes of HCV-resolvers following 1-2 weeks 

of HCV peptide stimulation or engineered by transducing CD8 

T cells from uninfected blood donors by lentivirus encoding 

HCV-TCR specific for HLA-A2 restricted NS3 1073- or NS5B 

2594-epitopes. HCV-specific or non-specific T cells were co-cultured 

with HCV-infected Huh7.5 cells at varying E/T ratio, culture 

duration and inhibitory receptor blockade, with the level


of HCV-expression in Huh7.5 cells confirmed by anti-NS5A Ab 

and HCV-specific T cell activation examined by MHC peptide 

tetramer, intracellular cytokine staining and CD107a mobilization 

using FACS. Result: Jc1-1073-1a and Jc1-2594-1a with 

genotype 1a-derived HCV epitopes could infect over 90% of 

Huh7.5 cells and support endogenous processing/presentation 

of 1a-based CD8 T cell epitopes to activate HCV-specific CD8 

T cells in-vitro. Co-culture with HCV-specific CD8 T cells and 

HCV-replicating Huh7.5 cells resulted in reduced number of 

total and HCV-infected Huh7.5 cells. PD-1 blockade enhanced 

HCV-specific CD8 T cell activation with reduced HCV expression 

as well as viability of Huh7.5 cells. Conclusion: PD-1 

blockade may enhance virus control through cytopathic pathway 

in an infectious HCV co-culture system, highlighting the 

need for caution in clinical application of checkpoint inhibitors. 

*acknowledgment: HCV and T-cell reagents were generously 

provided by Drs. Takaji Wakita, Charles Rice, Brett Lindenbach, 

Stanley Lemon, Jim Riley, Annelise Vuidepot, Peter Molloy 

and Alan Bennett. 

Disclosures: 

Kyong-Mi Chang - Consulting: Genentech, Tekmira, Alnylam; Stock Shareholder: 

BMS (spouse employment) 

The following authors have nothing to disclose: Keisuke Ojiro, Xiaowang Qu, 

Jang-June Park, Hyosun Cho 

1002 

HCV RNA and HCV core antigen are frequently detectable 

in stool of men chronically infected with HCV: Is 

feces a potential source of infection? 

Benjamin Heidrich 1,3 , Eike Steinmann 4 , Iris Plumeier 2 , Janina 

Kirschner 1 , Lisa Sollik 1 , Szilvia Ziegert 1 , Patrick Lehmann 1 , 

Michael Engelmann 4 , Tim Lankisch 1 , Michael P. Manns 1,3 , Dietmar 

H. Pieper 2,3 , Heiner Wedemeyer 1,3 ; 1 Dept. of Gastroenterology, 

Hepatology and Endocrinology, Hannover Medical School, Hannover, 

Germany; 2 Microbial Interactions and Processes Research 

Group, Centre for Infection Research, Braunschweig, Germany; 

3 Partner site Hannover-Braunschweig, German Center for Infection 

Research (DZIF), Hannover-Braunschweig, Germany; 4 Institute for 

Experimental Virology, TWINCORE Centre for Experimental and 

Clinical Infection Research, Hannover, Germany 

Background and Aims: HCV is one of the leading causes of 

liver cirrhosis and hepatocellular carcinoma worldwide. Transmission 

of HCV is thought to be mainly parenteral. However, 

unprotected anal intercourse seems to be a risk factor for acquisition 

of HCV in men having sex with men (MSM) as well as in 

heterosexual partnerships. HCV has been detected in blood, 

saliva, and bile. Surprisingly, there are no studies investigating 

the presence of HCV in stool. Methods: Stool samples of 98 

patients were prospectively collected, mixed with RNAlater 

and stored at -20°C. RNA was isolated using TRIzol reagent 

and transcribed in cDNA by using the SuperScript III kit with 

random hexamers. Specific HCV primers were used to identify 

samples positive for HCV RNA. PCR results were confirmed by 

Sanger sequencing. HCV RNA positive samples were tested for 

occult blood using the hemoCARE guajak test and were tested 

for HCVcoreAg using the Architect HCVAg from Abbott. Additionally, 

viral stability of recombinant HCV particles was investigated 

in vitro by incubation of the genotype 2a chimeric virus 

Jc1 with different bile and stool suspensions. Results: Overall, 

68/98 (69%) stool samples tested positive for HCV RNA. Men 

tested more often positive for HCV RNA in stool than women 

(83% vs. 52%; p


factor for HCV infection in Vero cells. We also supplemented 

Vero cells with the human Apolipoprotein E (ApoE), one of 

the host factors important for virus production, and found that 

ApoE supplementation enabled them to produce infectious 

viruses. Finally, we established Vero cells expressing all these 

factors, miR-122, human SR-B1 and ApoE. In this cell line, HCV 

replication and infectious virus production could be observed 

after HCVcc infection, indicating the reconstitution of the entire 

life cycle of HCV. In conclusion, we demonstrate that miR-122, 

SR-B1 and ApoE are necessary and sufficient for the reconstitution 

of the complete HCV life cycle in non-human non-hepatic 

Vero cells. These identified factors may be key determinants for 

species and organ tropism of HCV infection. The established 

novel HCV cell culture system with Vero cells will be useful to 

understand the species and organ specific restriction factors of 

HCV and to establish the mass culture system of viral particles 

in non-cancer cells for HCV vaccine. 

Disclosures: 

The following authors have nothing to disclose: Asako Murayama, Nao Sugiyama, 

Takaji Wakita, Takanobu Kato 

1004 

Tim-3 Inhibits Monocyte Functions via T-bet during Hepatitis 

C Virus Infection 

Ying Zhang 1 , Wenjing Yi 1 , Peixin Zhang 1 , Yan Liang 1 , Zhi Q. 

Yao 2 , Zhansheng Jia 1 ; 1 Department of Infectious Diseases, Tangdu 

Hospital, Fourth Military Medical University, Xi’an, China; 2 Department 

of Internal Medicine, Division of Infectious Diseases, James 

H. Quillen College of Medicine, East Tennessee State University, 

Johnson City, TN 

Hepatitis C virus (HCV) dysregulates innate immune responses 

and induces persistent viral infection. We have previously 

demonstrated that T-cell immunoglobulin and mucin domain 

protein-3 (Tim-3) plays a pivotal role in negative regulation 

of Toll-like receptor (TLR)-mediated innate immune responses. 

While it is clear that Tim-3 is up-regulated on monocyte/macrophages 

(M/M) during HCV infection, little is known about the 

transcription factor that controls its expression in these cells. 

Recent studies have revealed that Tim-3 expression is controlled 

by the transcription factor T-box expressed in T cells (T-bet). 

In this study, we further investigated the regulatory effects of 

T-bet on Tim-3 transcription/translation in M/M during HCV 

infection. Our results demonstrate that T-bet is constitutively 

expressed on resting peripheral blood CD14+ M/M. M/M 

from chronically HCV-infected subjects at baseline exhibit 

significantly increased T-bet expression that is positively correlated 

with the Tim-3 expression. Up-regulation of T-bet is also 

observed in CD14+ M/M incubated with HCV+ Huh7.5 cell in 

a time-dependent manner. In addition, HCV core protein can 

induce T-bet expression in primary M/M or monocytic THP-1 

cells, which is reversible by blocking the HCV core/gC1qR 

interactions. Moreover, the HCV core-induced up-regulation of 

T-bet and Tim-3 expression in CD14+ monocytes can be abrogated 

by incubating the cells with SP600125 - an inhibitor for 

JNK signaling pathway. Notably, silencing T-bet gene expression 

decreases Tim-3 expression and enhances IL-12 secretion 

as well as STAT-1 phosphorylation. These data suggest that 

T-bet, induced by HCV core/gC1qR through JNK pathway, 

enhances Tim-3 expression, leading to dampened M/M function 

during chronic HCV infection. These findings provide new 

information regarding Tim-3 transcriptional/translational regulation 

via T-bet during HCV infection and novel immunotherapy 

targets to combat this global epidemic viral infection. 

Disclosures: 

The following authors have nothing to disclose: Ying Zhang, Wenjing Yi, Peixin 

Zhang, Yan Liang, Zhi Q. Yao, Zhansheng Jia 

1005 

Downregulation of let-7 family miRs is strongly associated 

with increasing stages of hepatic fibrosis in chronic 

hepatitis C (HCV) Genotype 3a 

Manish C. Choudhary 1 , Sadaf Dar 1 , Ekta Gupta 2 , Jaswinder S. 

Maras 1 , Syed N. Kazim 3 , Gayatri Ramakrishna 1 , Shiv K. Sarin 1 ; 

1 Research, Institute of Liver and Biliary Science, New Delhi, India; 

2 Virology, Institute of Liver and Biliary Sciences, New Delhi, India; 

3 Centre for Interdisciplinary Research in Basic Sciences, Jamia 

Milia Islamia, New Delhi, India 

Background: Chronic hepatitis C (CHC) infection is a major 

cause of liver fibrosis and end stage liver disease. Circulating 

miRNAs have evolved as a reliable biomarker for various 

pathological conditions. However, there is limited data on systematic 

miRNA based biomarker study on hepatic fibrogenesis 

in HCV genotype 3a (G3a). Aims: To profile circulating repertoire 

of miRNAs in the plasma of HCV G3a infected patients 

with different stages of hepatic fibrosis.Patients and Methods: 

47 subjects with CHC and histological assessment by two independent 

hepato-pathologists were categorized based on stage 

of hepatic fibrosis: F0-1 (n=32), F3-4 (n=15) were compared 

with healthy controls (n=28). Differentially expressed miRNAs 

in plasma of CHC F0,1 (n=4) and F3,4 (n=4) were studied 

and compared with controls using miRCURY LNA TM Serum/ 

Plasma Focus panel. These miRNAs were analysed in context 

of fibrosis progression by making comparison between CHC 

F0,1 and F3,4 patients. Subsequent validation of array data 

was done by stem-loop RT-PCR. miRNA target gene prediction 

was done by multiMiR R analysis. Results: 185 miRNAs were 

screened and 31 miRNAs were commonly identified in all the 

groups. 4 miRNA were significantly down regulated (p


1006 

Type III interferon responses contribute to hepatitis virus 

infection and depended on IFNL3-IFNL4 variant 

Tsunamasa Watanabe 2,1 , Susumu Tsutsumi 2 , Kentaro Matsuura 3 , 

Etsuko Iio 3 , Noboru Shinkai 3 , Kayoko Matsunami 3 , Fumio Itoh 1 , 

Yasuhito Tanaka 2 ; 1 Department of Internal Medicine, Division of 

Gastroenterology and Hepatology, St. Marianna University School 

of Medicine, Kawasaki, Japan; 2 Department of Virology and Liver 

Unit, Nagoya City University Graduate School of Medical Sciences, 

Nagoya, Japan; 3 Department of Gastroenterology and 

Metabolism, Nagoya City University Hospital, Nagoya, Japan 

Introduction: Type III interferon, which consist of interferon-l 

(IFNL)1, 2, 3, and 4, has received considerable attention in 

hepatotropic hepatitis C virus (HCV), as many independent 

genome-wide association studies have identified a strong association 

between IFNL3-IFNL4 variants and outcome of HCV. 

Materials and Methods: We employed an original primary 

human hepatocytes (PHH) systems, isolated from chimeric 

mice harboring human hepatocytes without cryopreservation, 

to investigate innate immune responses against hepatotropic 

viruses including mimic of HCV infections and hepatitis B virus 

(HBV) infection. The antiviral effects of IFNL were also investigated 

in vivo using chronically HCV-infected chimeric mice 

transplanted with human hepatocytes of IFNL3-IFNL4 favorable 

or unfavorable genotype. Results: In vivo study using pegylated 

IFNL1 revealed that the initial viral decline of HCV-RNA by 

IFNL1 was similar in both IFNL3-IFNL4 favorable and unfavorable 

genotype. Compared to the effect of pegylated IFN-a, 

however, the antiviral effects of IFNL was limited, because of 

the killer activity of NK cells that was predominantly induced 

by only IFN-a, not IFNL1. On the other hands, type III IFN 

responses using PHH culture in vitro against 5’-triphosphate 

single stranded RNA (PPP-RNA) transfection (mimic of HCV 

infection) were varied from IFNL3-IFNL4 variants, although the 

levels of IFN-λ genes were significantly higher in PHHs with 

favorable genotype than that with unfavorable genotype by 

PPP-RNA. The knockdown experiment by small interfering RNA 

indicated that cytosol RNA sensor was the key molecule of IFNL 

production. When PHHs were infected with HBV, IFN-λ genes 

were induced after HBV infection in PHHs with favorable genotype, 

but not unfavorable genotype. Conclusions: Our results 

suggest that a response of human hepatocytes against hepatotropic 

viruses could be contribute to type III IFN production and 

the responses might be depended to the IFNL3-IFNL4 variants. 

Disclosures: 

Yasuhito Tanaka - Grant/Research Support: Chugai Pharmaceutical CO., LTD., 

MSD, Bristol-Myers Squibb; Speaking and Teaching: janssen pharma, Bristol-Myers 

Squibb 

The following authors have nothing to disclose: Tsunamasa Watanabe, Susumu 

Tsutsumi, Kentaro Matsuura, Etsuko Iio, Noboru Shinkai, Kayoko Matsunami, 

Fumio Itoh 

1007 

Altered frequencies and function of MAIT cells during 

treatment of patients with chronic HCV, HIV and HCV/ 

HIV co-infections 

Michelle Spaan 2 , Sebastiaan Hullegie 1 , Kim Kreefft 2 , Gertine van 

Oord 2 , Boris Beudeker 2 , Bart J. Rijnders 1 , Robert J. de Knegt 2 , 

Mark Claassen 1 , Andre Boonstra 2 ; 1 Department of Internal Medicine, 

Erasmus MC, Rotterdam, Netherlands; 2 Department of 

Gastroenterology and Hepatology, Erasmus MC, Rotterdam, Netherlands 

Background Mucosal invariant T (MAIT) cells comprise a subpopulation 

of T cells that can be activated by a broad range of 

bacterial products and cytokines to produce IFN-γ and express 

cytolytic enzymes. Loss of MAIT cells is observed in HIV infection, 

which is thought to compromise anti-bacterial immunity 

and microbial translocation in the gut. Since little is known 

on the MAIT cells during HCV infection, we compared their 

phenotype and function in comparison to HIV and HCV/ HIV 

co-infection, and determined the effect of IFN-based and DAA 

antiviral therapy on MAIT cells for HCV. Methods Peripheral 

blood was collected before, during and 24 weeks after therapy 

from patients with chronic HCV (n=27), virologically supressed 

chronic HIV (n=10), HCV/HIV co-infection (n=9) and healthy 

individuals (n=12) before, during and after therapy. Patients 

with HCV were treated with pegIFN-α/ribavirin with and without 

telaprevir, or boceprevir or with IFN-free therapy. MAIT 

cells were identified on the basis of CD161 and Vα7.2, and 

were stimulated with E.coli, IL-12/IL-18 or IL-18/IFN-α for 18 

hours. NK cells and MAIT cells were analysed for the expression 

of CD38, CD69 and IFN-γ by flowcytometry. Results Compared 

to healthy individuals, the frequency of MAIT cells was 

significantly decreased in patients with chronic HCV, HIV and 

HCV/ HIV co-infection (5.6%, 3.2%, 2.0% and 1.0% within 

CD3+ T cells, respectively). Expression of CD38 on MAIT cells 

was comparable in chronic HCV, HIV and healthy individuals, 

but was significantly increased in patients with HIV/HCV 

co-infection. MAIT cells from healthy controls and the 3 patient 

populations were responsive to IFNα in vitro as evidenced by 

enhanced frequencies of CD69 expressing and IFN-γ producing 

cells. After successful IFN-based therapy the frequencies of 

MAIT cells did not normalize. However, we observed that the 

functionality of MAIT cells was differentially affected during the 

course IFN-α-based therapy and was highly dependent on the 

treatment regimen and patient group. Importantly, viral load 

decline by IFN-free DAA treatment led to strongly enhanced 

IFN-γ levels. Conclusion We show that the frequencies of MAIT 

cells are reduced in blood of patients with chronic HCV, HIV 

and in HCV/HIV co-infection compared to healthy individuals. 

The potent effects on MAIT cells of exposure to IFN-α, both in 

vitro and in vivo, and HCV and HIV RNA levels warrant more 

detailed studies on the interplay between infections to the activity 

of this specialized T cell subpopulation. 

Disclosures: 

Bart J. Rijnders - Advisory Committees or Review Panels: BMS, Abbvie, Gilead; 

Grant/Research Support: MSD, Gilead 




Andre Boonstra - Grant/Research Support: BMS, Janssen Pharmaceutics, Merck, 

Roche, Gilead 

The following authors have nothing to disclose: Michelle Spaan, Sebastiaan Hullegie, 

Kim Kreefft, Gertine van Oord, Boris Beudeker, Mark Claassen 

1008 

Interferon-based treatment activates regulatory CD4 + 

T-cells to exert counteractive antiviral immunity 

Bettina Langhans, Hans Dieter Nischalke, Philipp Lutz, Benjamin 

Krämer, Christian P. Strassburg, Jacob Nattermann, Ulrich Spengler; 

Department of Internal Medicine I, University of Bonn, Bonn, 

Germany 

Background and aims: Regulatory CD4 + T cells (Tregs) accumulate 

during chronic hepatitis C virus (HCV) infection and 

inhibit anti-viral T cell responses. The availability of direct-acting 

antiviral agents (DAAs), which can be applied in combination 

with pegylated interferon (IFN) or as IFN-free regimens 

have revolutionized treatment options. However, their effect on 

altered immunoregulation in hepatitis C is unclear. Here, we 

studied Tregs before and after elimination of HCV with DAAs 

combined with IFN versus IFN-free therapy. Methods: Using


multi-color flowcytometry we analyzed frequency and function 

of Tregs and effector T cells before and at the end of treatment 

(treatment week (TW) 12) from HCV-patients after succesful 

HCV elimination with sofosbuvir (SOF) in combination with 

IFN plus ribavirin (n=14) versus patients treated with SOF plus 

daclatasvir (DCV) and simeprevir (SMV), respectively (n=14). 

In addition, we measured serum levels of immunomodulatory 

cytokines. Results: Unlike IFN-free therapy, treatment with SOF 

plus IFN significantly increased numbers of peripheral Tregs 

(% Foxp3 + CD25 + CD4 + T cells TW0 vs. TW12 (MW±SD): 

5.5±2.0 vs. 7.2±2.4; p90%) of sustained 

virological response at week 12 (SVR12) in patients with 

HCV infection genotype 4 (Abergel et al. ILC 2015, J Hepatol 

2015; 62 (Suppl 2):S219; Poordad et al. ILC 2015, J Hepatol 

2015; 62 (Suppl 2):S261). In the former study, two of the three 

patients with virological relapse had a genotype 4r subtype. 

In the later, NS5A variants were found at baseline in 4 out of 

13 patients with virologic failure and in 13 out of 13 patients 

at failure. In addition, the number of pre-treatment baseline 

RAPs (Resistance Associated Polymorphisms) was associated 

with virological failure in a study conducted by Sarrazin et 

al. (AASLD, Hepatology 2014; 60 (Suppl) 1128A). Aim: The 

objective of this study was to assess the NS5A natural polymorphisms 

of the genotype 4 subtypes in order to investigate 

the potential role of subtypes and RAPs in treatment outcome 

with regimens containing an NS5A inhibitor. Methods: We 

collected from the NCBI, European, and Japanese HCV databases, 

47 HCV NS5A Genotype 4 sequences and proceeded 

to multiple Sequence Alignment using Clustal W2. We analyzed 

16 subtype 4a, 7 subtype 4b, 3 subtype 4d, 2 subtype 

4m, 3 subtype 4o and 16 subtype 4r (no 4c or 4f were available). 

We assessed the prevalence by subtypes of the main 

RAPs: L28M, L30R, L31M, P58T and Y93H. Results: Seventy 

five percent of the 16 genotype 4r strains included 2 RAPs 

(L30R and L31M), in comparison with the 16 genotype 4a 

which had only one RAP in all of the strains included (L31M) 

(p


the innate immune response following Poly(I:C) and HCV-RNA 

transfection. LECT2 enhanced RIG-I-mediated IFNβ induction 

and increased ISG induction through the activation of IRF3, 

IRF7 and JAK/STAT pathway. To confirm the biologic significance 

of these results, we generated LECT2 transgenic (Tg) 

and knockout (KO) mice. LECT2-Tg mice showed increased ISG 

induction following Poly(I:C) treatment in the liver. Conversely, 

LECT2-KO mice showed impaired ISG induction following IFN 

or Poly(I:C) treatment to 30–50% of that of WT mice. Conclusion: 

Gene expression profiling of primary hepatocytes treated 

with IFNα, IL28B, or both in combination revealed that LECT2, 

which enhances the innate immune response and suppresses 

HCV replication, was specifically induced by IL28B. LECT2 

might participate in prolonged ISG induction by IL28B and in 

the unique innate antiviral immune system of IL28B. 

Disclosures: 


Stanley M. Lemon - Advisory Committees or Review Panels: Merck, Santaris, 

Abbott, Gilead; Consulting: Achillion, Idenix; Grant/Research Support: Merck, 

Tibotec, Scynexis; Speaking and Teaching: Hoffman LaRoche 






The following authors have nothing to disclose: Takayoshi Shirasaki, Masao 

Honda, Tetsuro Shimakami, Kazuhisa Murai, Hirofumi Misu, Toshinari Takamura, 

Seishi Murakami 

1011 

MicroRNA 17 Host Gene Protein and its targets PTEN 

and NF-κB in Chronic Hepatitis C Virus Infection: Relation 

to Hepatic inflammation, fibrosis and steatosis. 

Hoda El Aggan 1 , Sahah A. Mahmoud 2 , Nevine M. El Deeb 3 , 

Ehab M. Hassona 1 , Sally El Demiry 1 ; 1 Department of Medicine 

(Hepatobiliary Unit), Faculty of Medicine, University of Alexandria, 

Alexandria, Egypt; 2 Department of Medical Biochemistry, Faculty 

of Medicine, University of Alexandria, Alexandria, Egypt; 3 Department 

of Pathology, Faculty of Medicine, University of Alexandria, 

Alexandria, Egypt 

Background/Aim: MicroRNAs (miRs) regulate post-transcriptional 

gene expression in various biological processes. The 

polycistronic miR-17~92 cluster is comprised of six miRs and 

its primary transcript also encodes for a polypeptide of 70 

amino acids designated as the miR-17 host gene (MIR17HG) 

protein. The present study was designed to evaluate the plasma 

levels of MIR17HG protein, an index of miR-17~92 cluster 

activity, and hepatic expression of its targets phosphatase and 

tensin homolog (PTEN) and nuclear factor kappa-B (NF-κB) 

in patients with chronic hepatitis C virus (HCV) infection in 

relation to hepatic steatosis, inflammation and fibrosis. Methods: 

Thirty treatment-naïve patients with chronic HCV infection 

[18 patients with chronic hepatitis C (CHC) and 12 patients 

with cirrhosis] and 15 healthy subjects were included in the 

study. Quantitative determination of plasma levels of MIR17HG 

protein was performed using quantitative sandwich enzyme 

immunoassay. Core liver biopsies obtained from patients with 

chronic HCV infection were assessed for METAVIR histological 

activity grade and fibrosis stage and steatosis grade. Immunohistochemical 

staining of liver specimens was done using 

monoclonal antibody against PTEN and NF-κB. Results: Plasma 

MIR17HG protein levels showed a significant increase in 

patients with chronic HCV infection compared with healthy subjects 

(P = 0.012) and in patients with cirrhosis compared with 

patients with CHC (P < 0.001). By plotting a receiver-operating 

characteristic curve, the sensitivity and specificity of plasma 

MIR17HG protein levels in discriminating patients with cirrhosis 

from patients with CHC were 100% and 88.9% respectively 

at a cut-off level of 45.5 pg/ml (AUC = 0.995). The plasma 

MIR17HG protein levels were inversely correlated with hepatic 

PTEN expression and positively correlated with serum levels of 

aminotransferases, METAVIR histological activity grade and 

fibrosis stage, steatosis grade and hepatic NF-κB expression 

(P < 0.05). The hepatic PTEN expression showed positive correlations 

with serum gamma glutamyl transpeptidase levels, 

METAVIR fibrosis stage and steatosis grade (P < 0.05) and an 

inverse correlation with hepatic NF-κB expression (P = 0.006). 

Conclusion: Activation of the miR-17~92 cluster may play an 

important role in the pathogenesis of HCV-related liver injury 

via PTEN inhibition and NF-κB activation and could be a possible 

therapeutic option in chronic HCV infection. Circulating 

MIR17HG protein could be a useful biomarker for disease 

progression. 

Disclosures: 

The following authors have nothing to disclose: Hoda El Aggan, Sahah A. Mahmoud, 

Nevine M. El Deeb, Ehab M. Hassona, Sally El Demiry 

1012 

The FGL2:FcγRIIB Immunosuppressive Pathway Inhibits 

HCV Specific Antiviral T Cell Responses in Chronic Infection 

Ramzi Khattar, Kaveh Farrokhi, Hassan Sadozai, Vanessa Rojas 

Luengas, Gary Levy, Nazia Selzner; University Health Network, 


Background and Aims: Hepatitis C virus (HCV) evades immune 

detection by limiting the magnitude and responsiveness of antiviral 

T cells. FGL2 is a potent immunosuppressive effector molecule 

of CD4 + CD25 + TIGIT + FOXP3 + Treg that exerts its activity 

by binding to the inhibitory FCγRIIB receptor expressed on antigen 

presenting cells. Binding of FGL2 to the FCγRIIB receptor 

inhibits the maturation of dendritic cells, limiting responsiveness 

of antiviral T cells. We examined the relationship between 

FGL2 and HCV specific T cell responses in a cohort of HCV 

patients. The study also examined the efficacy of a novel neutralizing 

antibody towards FGL2 to restore HCV specific T cell 

proliferation in vitro from HCV patients. Methods: 61 patients 

with chronic HCV infection, 15 SVR and 8 healthy controls 

were recruited to the study. TIGIT expression on CD4+CD25+- 

FOXP3+ Treg and FGL2 production by T cells, dendritic cells 

and macrophages was measured in patient PBMC by flow 

cytometry. FGL2 levels were measured in patient samples by 

ELISA and T cell reactivity to HCV by ex vivo peptide re-stimulation 

using genotype specific HCV peptide arrays. Results: In 

patients chronically infected with HCV, there was a significant 

increase in frequency of CD4 + CD25 + FOXP3 + TIGIT + Treg cells 

(42.5±4.1) compared to SVR patients (30.1±2.8) or healthy 

controls (14.5±1.5) (P


Disclosures: 

The following authors have nothing to disclose: Ramzi Khattar, Kaveh Farrokhi, 

Hassan Sadozai, Vanessa Rojas Luengas, Gary Levy, Nazia Selzner 

1013 

Novel effect of NS5A inhibition on Augmentation of 

HCV specific immunity and SVR 

Shikha Shrivastava 1 , Anu Osinusi 2 , Shyam Kottilil 1 ; 1 Institute of 

Human Virology, University of Maryland, Baltimore, Baltimore, 

MD; 2 Gilead Sciences Inc,, Foster City,, CA 

Background: We previously demonstrated that HCV clearance 

is associated with increased HCV specific immunity in 

chronic hepatitis C virus (HCV) genotype 1 (GT-1) infected 

patients during treatment with the Interferon (IFN) free regimen 

of sofosbuvir and ribavirin (SPARE). In this study, we 

evaluated the HCV specific immune responses in patients who 

relapsed previous treatment with sofosbuvir and ribavirin but 

were retreated with sofosbuvir and ledipasvir. Aim: To analyze 

the changes in HCV specific immunologic responses associated 

with viral clearance with combination DAA therapy of 

Sofosbuvir and Ledipasvir in chronic HCV GT1 patients who 

relapsed and lacked enhancement of HCV specific immune 

response when treated with sofosbuvir and ribavirin (SPARE). 

Methods: HCV GT-1 patients (N=14) that relapsed after treatment 

with sofosbuvir and ribavirin for 24 weeks were retreated 

with Sofosbuvir and Ledipasvir for 12 weeks. Phenotypic and 

functional changes within the adaptive and innate immune 

compartments of peripheral blood mononuclear cell (PBMC) 

at baseline and end of treatment (EOT) were analyzed. HCV 

specific immune responses were quantified by ELISA, ELISpot 

and intracellular flow cytometry (IFN-gamma, IL-2, TNF-a) after 

stimulation with overlapping peptides spanning the entire HCV 

genome. Results: In the SPARE study, SVR was associated with 

an enhancement of HCV-specific immune response (IFN-G positive 

peripheral CD8+ T cells). During the retreatment of these 

relapsers from SPARE with Sofosbuvir and Ledipasvir for 12 

weeks, all patients have attained SVR 12. Suppression of HCV 

was associated with decline in T cell exhaustion markers at EOT 

such as CD8+CD57+ (9.3 + 0.2 % vs. 6.2+ 0.6% p=0.02), 

CD8+Tim3+ (67+10.2 % VS. 51+9.3 % p=0.03), CD8+PD1+ 

(17+1.8 % VS. 12+1.5% p=0.03). All patients were able to 

augment peripheral HCV-specific T cell IFN-gamma responses 

at the end of treatment compared to baseline when treated with 

sofosbuvir and ledipasvir (p=0.003). SVR was associated with 

augmentation of HCV-specific T cells response. Conclusions: 

Addition of NS5A inhibitor to sofosbuvir is associated with 

augmentation of HCV specific immunity and SVR in patients 

who previously failed sofosbuvir and ribavirin therapy. These 

findings demonstrate a novel effect of NS5A inhibitors in inducing 

host response aiding HCV clearance and achieving SVR. 

Disclosures: 


The following authors have nothing to disclose: Shikha Shrivastava, Shyam Kottilil 

1014 

Differential microRNA expression in the liver during the 

advanced stage of chronic hepatitis C 

Rika Horii, Masao Honda, Takayoshi Shirasaki, Hikari Okada, 

Mikiko Nakamura, Shuichi Kaneko; Kanazawa University Graduate 

School of Medicine, Kanazawa, Japan 

BACKGROUND & AIMS: MicroRNAs (miRNAs) play important 

roles in development, metabolism, infection, and cancer. 

We previously analyzed the changes of miRNA expression 

associated with the progression of chronic hepatitis C (CHC) 

and showed that the expression of 7 miRNAs (miR-10a, -19a, 

-27a, -195, -199a, -214, and -218) were most significantly 

different between patients with early stage fibrosis (F1–2) and 

advanced stage fibrosis (F3–4). In this study, we evaluated 

the functional relevance of these miRNAs on Hepatitis C virus 

(HCV) replication and pathogenesis of liver disease. METH- 

ODS: MiRNAs were obtained from biopsy specimens of CHC 

patients infected with genotype 1b HCV. The functional relevance 

of miRNAs on HCV replication was evaluated in Huh-7 

cells using the infectious genotype 1a clone pH77S.3/Gluc2A 

with a Gaussia reporter gene. HCV translation (HCV-IRES) was 

monitored in the stably transformed IRES reporter cell line RCF- 

26. MiRNA-transfected Huh-7 cells were infected with HCV 

particles in oleic acid- or chenodeoxycholic acid (CDCA)–containing 

medium and HCV replication and cellular metabolism 

were evaluated. RESULTS: Among 7 miRNAs differentially 

expressed between early and advanced stage fibrosis (miR- 

10a, -19a, -27a, -195, -199a, -214, and -218), most miRNAs 

other than miR-19a were significantly up-regulated in Huh-7 

cells following HCV infection. Conversely, overexpression of 

miR-10a, -27a, -195, -199, and -214 significantly repressed 

HCV replication in Huh-7 cells, while miR-19a and -218 had 

no effect on HCV replication. We focused on miR-10a, as 

it was not previously characterized in HCV replication and 

liver pathogenesis. MiR-10a had no effect on HCV translation; 

however, its overexpression decreased hepatic triglyceride synthesis 

by suppressing lipid synthetic genes such as SREBP1 and 

FASN. Moreover, CDCA treatment in HCV replicating Huh-7 

cells increased hepatic concentrations of cholesterol but miR- 

10a decreased hepatic cholesterol content and suppressed 

HCV replication. We found that miR-10a targeted liver receptor 

homolog-1, a key regulatory enzyme of bile acid biosynthesis 

from cholesterol 7α-hydroxylase/CYP7A1. Thus, miR-10a 

suppresses the storage of triglyceride and cholesterol and the 

synthesis of bile acid, suppressing HCV replication. This would 

protect liver cell damage in the advanced fibrosis stage of liver 

disease. Conclusion: MiR-10a possibly plays an important role 

in cellular lipid and bile acid metabolism. The findings of this 

report might serve to establish a new therapeutic strategy for 

HCV-related advanced liver disease. 

Disclosures: 







The following authors have nothing to disclose: Rika Horii, Masao Honda, Takayoshi 

Shirasaki, Mikiko Nakamura 

1015 

The tumor suppressor IQGAP2 plays an essential role 

in antiviral activity of Interferon alpha against HCV 

through the NF-κB pathway. 

Cynthia Brisac, Shadi Salloum, Victor Yang, Stephane Chevaliez, 

Esperance A. Schaefer, Jian Hong, Charles Carlton-Smith, Nadia 

Alatrakchi, Anna Lidofsky, Dahlene Fusco, Xiao Liu, Dachuan Cai, 

Lee F. Peng, Wenyu Lin, Raymond T. Chung; MGH, Boston, MA 

Type I Interferon (IFN) has been the backbone of treatment for 

hepatitis C virus (HCV) infection over the past two decades. For 

some countries, it is likely to remain the only treatment since 

they will not be able to afford new high cost IFN-free regimen. 

Recently, using unbiased genome-wide siRNA screens, we 

identified hundreds of novel genes required for IFN response 

against HCV (Zhao H, J. Hepatology, 2013 and Fusco DN,


Gastroenterology, 2014) shedding light on new pathways of 

IFN. It is now clear that JAK/STAT and the hundreds of Interferon 

Stimulated Genes (ISGs) it stimulates is not the only pathway 

mediating IFN’s antiviral effects in the liver. One of the 

genes we identified is IQ-motif containing GTPase activating 

protein 2 (IQGAP2). IQGAP2 is predominantly expressed in 

the liver and has been described as a tumor suppressor. Here 

we show that IQGAP2 also mediates IFN anti-HCV response 

in infected hepatoma cells. Indeed, IQGAP2 siRNA-mediated 

knock-down rescues HCV infection from IFN inhibition. We 

show that IQGAP2 is not itself an ISG and its knockdown 

does not affect JAK/STAT activation, suggesting that IQGAP2 

acts independently of the JAK/STAT pathway. We found that 

IQGAP2 interacts with RelA/p65, a subunit of NF-κB transcription 

factor. Interestingly, our data suggest that IFN alone 

is sufficient to activate RelA and stimulate NF-κB-dependent 

transcription in hepatoma cells; this process requires IQGAP2 

expression. Furthermore, we found that RelA silencing mimics 

IQGAP2 knock-down effects in rescuing HCV from IFN inhibition 

in IFN-deficient cells. This suggests that IQGAP2 and RelA 

mediate IFN anti-HCV response downstream of IFN binding to 

its receptor, a mechanism different from the well characterized 

role of NF-κB in IFN production. Finally, we investigated the 

effect of IQGAP2 or RelA knock-down on the induction by 

IFN of 23 ISGs with previously described anti-HCV properties. 

We found that IQGAP2 and RelA are both required for the 

full induction of two thirds of the tested genes suggesting that 

IQGAP2 and RelA mediate IFN anti-HCV response by controlling 

ISG induction in parallel of the JAK/STAT pathway. 

Altogether, our data demonstrate a previously unrecognized 

function for IQGAP2 in regulating innate antiviral response 

against HCV, and probably other hepatotropic pathogens. 

Since IQGAP2 is predominantly expressed in the liver, understanding 

the mechanism of RelA activation by IQGAP2 may 

open novel opportunities to prevent NF-kB associated liver disease. 

Disclosures: 

Stephane Chevaliez - Advisory Committees or Review Panels: Janssen; Speaking 

and Teaching: Gilead, BMS 



Merck, BMS 

The following authors have nothing to disclose: Cynthia Brisac, Shadi Salloum, 

Victor Yang, Esperance A. Schaefer, Jian Hong, Charles Carlton-Smith, Nadia 

Alatrakchi, Anna Lidofsky, Xiao Liu, Dachuan Cai, Lee F. Peng, Wenyu Lin 

1016 

Single-strand RNA-induced monocyte differentiation 

generates pro-fibrogenic M2 macrophages in chronic 

hepatitis C Virus infection 

Banishree Saha, Karen Kodys, Gyongyi Szabo; Medicine, UMASS 

Med School, Worcester, MA 

Purpose: Innate immune responses, including monocyte and 

macrophage activation, contribute to the pathogenesis of 

chronic Hepatitis C Virus (HCV) infection and liver fibrosis. 

However the mechanisms that lead to liver fibrosis during 

HCV infection are not well understood. In response to the 

tissue microenvironment, monocytes undergo differentiation 

into polarized states including M1 (pro-inflammatory) or M2 

(anti-inflammatory/pro-fibrotic) macrophages. We hypothesized 

that during chronic HCV infection monocytes undergo 

differentiation that affect liver disease progression. Methods: 

Healthy human monocytes (n=10-15) were cultured with cellfree 

HCV or exosomes isolated from HCV-infected Huh7.5 

hepatoma cells for 5-7 days. Circulating monocytes and liver 

macrophages from HCV-infected patients and controls were 

analyzed by flow cytometry and western blotting. Results: 

Co-culture of healthy monocytes with cell-free HCV or exosomes 

derived from HCV-infected Huh7.5 cells resulted in differentiation 

of monocytes into macrophages (MΦ) with high CD14 

and CD68 expression. Exosomes from HCV-infected Huh7.5 

cells contained HCV single stranded (ss) RNA. These MΦ displayed 

M2 markers (CD206, CD163 and DC-SIGN) and produced 

pro- and anti-inflammatory cytokines. The HCV-induced 

M2-MΦ led to hepatic stellate cell (LX2) activation indicated 

by increased expression of collagen, TIMP-1 and α-SMA and 

this was prevented by anti-TGFβ blocking antibody administration. 

The HCV-induced monocyte activation and differentiation 

into M2-MΦ was prevented by TLR8 but not TLR3 or 

TLR7 knock-down. TLR8 ligands, independent of HCV, caused 

monocyte differentiation and M2-MΦ polarization suggesting 

a role for TLR8 in this process. Furthermore, HCV ssRNA alone 

induced monocyte to M2-MΦ differentiation and polarization, 

demonstrating that HCV ssRNA signals via TLR8 induce monocyte 

differentiation and polarization. In vivo in patients with 

chronic HCV infection, we observed a significant increase in 

the expression of M2 markers (CD206, CD163) on circulating 

monocytes and in the liver. Furthermore, we found a significant 

increase in collagen + CD206 + CD14 + circulating monocytes 

and the increased frequency of collagen + CD206 + CD14 + circulating 

monocytes correlated with liver fibrosis in HCV-infected 

patients. Conclusion: Our data identify a new mechanism by 

which cell-free and exosome-packaged HCV ssRNA interacts 

with monocytes and induces TLR8-mediated differentiation to a 

pro-fibrogenic, M2-MΦ. These M2-MΦ promote liver fibrosis. 

We also identified M2-marker plus collagen-expressing circulating 

monocytes as potential biomarkers of fibrosis in chronic 

HCV infection. 

Disclosures: 

The following authors have nothing to disclose: Banishree Saha, Karen Kodys, 

Gyongyi Szabo 

1017 

Elucidating Mechanisms Underlying Development of 

Liver Disease in HIV/HCV Coinfection 

Jinhee Hyun 3 , Masato Yoneda 1 , Eugene R. Schiff 1 , Emmanuel 

Thomas 1,2 ; 1 Schiff Center for Liver Diseases, University of Miami 

Miller School of Medicine, Miami, FL; 2 Sylvester Comprehensive 

Cancer Center, Miami, FL; 3 Cell Biology, University of Miami 

School of Medicine, Miami, FL 

BACKGROUND: Patients with HIV that are coinfected with 

HCV are at increased risk for rapidly progressive liver disease 

and subsequently the development of Hepatocellular Carcinoma 

(HCC). Specifically, HCC develops earlier in coinfected 

patients and these patients are more symptomatic than those 

with only HCV infection at diagnosis suggesting that both viruses 

increase the propensity for malignant transformation. However, 

the genetic and cellular based mechanisms underpinning how 

HCV initiates and subsequently induces liver pathology and 

why coinfection with HIV results in significantly worse hepatic 

disease remains to be clarified. In addition, the specific cell 

types that contribute to these clinical outcomes are unknown. 

In this study, we specifically are testing the hypothesis that the 

robust viral RNA dependent innate immune response that we 

have characterized, that drives inflammation in HCV infected 

livers, is augmented with HIV coinfection through activation 

of viral DNA and RNA sensing pathways. METHODS: To this 

end, we have developed novel in-vitro models that utilize HCV 

infected primary human hepatocytes (PHHs) and HIV infected 

primary kupffer cells (PKCs). Specifically, the JFH-1 strain was 

used to study HCV while the BAL (R5) and IIIB (X4) strains were


used for HIV infection. A coculture model of PHHs an PKCs was 

also utilized. Specific RNA and DNA dependent innate and 

inflammatory pathways, such as RIG-I/TLR3 and IFI16/cGAS 

signaling, were studied as well as additional pathways involving 

the production of interferon and chemokines such as IP10/ 

CXCL10, IL-1b and CCL5/Rantes that may drive subsequent 

liver disease. RESULTS: We found that similar to distinct stimuli 

including IFN-a and polyI:C, HCV and HIV stimulation of these 

primary liver cells resulted in rapid and robust upregulation of 

IP10 and other inflammatory cytokines at the mRNA and protein 

level. Surprisingly, microarray analyses of HIV treated liver 

cells demonstrated several innate immune pathways were activated 

by this virus in the liver. siRNA experiments demonstrated 

the induction of inflammatory cytokines was mediated by IRF3 

and NFkB-dependent pathways. CONCLUSIONS: Overall, 

our data demonstrate that the robust antiviral response that is 

observed in HCV infected livers and subsequently drives liver 

pathology is augmented with HIV/HCV coinfection. It is hoped 

that understanding the mechanism by which coinfected patients 

have more severe liver disease will enable the development of 

targeted therapeutics to abrogate these poor clinical outcomes. 

Disclosures: 






The following authors have nothing to disclose: Jinhee Hyun, Masato Yoneda, 

Emmanuel Thomas 

1018 

Expression of IFNλ4 in liver is closely associated with 

non-response to antiviral therapy through the regulation 

of basal expression of ISGs in chronic hepatitis C 

patients but not in hepatitis B patients. 

Miyako Murakawa 1,3 , Yasuhiro Asahina 1 , Fukiko Kawai-Kitahata 

1 , Hiroko Nagata 1 , Syun Kaneko 1 , Sayuri Nitta 1 , Takako 

Watanabe 1 , Yasuhiro Itsui 1,3 , Mina Nakagawa 1 , Sei Kakinuma 1 , 

Sayuki Iijima 2 , Yasuhito Tanaka 2 , Mamoru Watanabe 1 , Yujiro 

Tanaka 3,1 ; 1 Gastroenterology and Hepatology, Tokyo Medical 

and Dental University, Tokyo, Japan; 2 Nagoya City University, 

Nagoya, Japan; 3 Medical Education Research and Development, 

Tokyo Medical and Dental University, Tokyo, Japan 

Backgrounds & Aims: The innate immune system has an essential 

role in host antiviral defense. IL28B SNPs are associated 

with the response to anti-HCV therapy both in IFN-based and 

IFN-free regimens. Recently, IFNλ4 gene was considered as 

one of the mechanisms responsible for poor response influenced 

by IL28B unfavorable SNPs. However, little is known 

about IFNλ4 expressions and its roles in antiviral innate immunity 

including intrahepatic gene expressions of IFN-stimulated 

genes (ISGs). The aim of this study is to evaluate the relationship 

between IFNλ4 expression and viral liver diseases, and to 

investigate the effect of IFNλ4 on intrahepatic gene expressions 

responsible for poor response to IFN therapy. Methods: 1) 

IFNλ4 expressions were analyzed in HCVcc-infected Huh7.5.1 

cells, poly (I:C)-treated Huh7, HepG2, HeLa, HEK293T cells 

and B lymphocytes, and HBV-expressing HepG2.215 cells. 

2) The expressions of IFNλ4 were analyzed in liver samples 

obtained from 49 chronic hepatitis C (CHC), 13 chronic hepatitis 

B (CHB) and 3 autoimmune hepatitis (AIH) patients. All 

CHC patients were treated with PEG-IFN/ribavirin (PR) therapy 

and the relationship between IFNλ4 expressions and 

treatment responses. 3) Antiviral ISGs (ISG15, Mx1, RIG-I, 

TLR3, SOCS2) and IFN-suppressive genes (RNF125, SOCS1, 

SOCS3, A20, RNF11) were analyzed in all liver samples, 

and the relationship with IFNλ4 expressions were accessed. 

Results: 1) IFNλ4 expression was induced by poly (I:C) treatment 

as well as HCVcc infection in IL28B-minor cell lines, but 

not induced by HBV. 2) IFNλ4 mRNA was detected in 11 of 

22 liver samples of CHC patients with IL28B-unfavorable SNP, 

but not in CHC patients with favorable genotype, CHB and AIH 

patients. IFNλ4 expression was associated with non-response 

to PR therapy (p < 0.001). 3) Intrahepatic expressions of antiviral 

ISGs (ISG15 and Mx1) were significantly up-regulated in 

nonvirological responders (NR) compared with those in virological 

responders (VR) while expressions of suppressive ISGs 

(RNF125, SOCS1, SOCS3 and RNF11) were down-regulated 

in NR. Expressions of antiviral ISGs (Mx1 and RIG-I) were significantly 

higher in IL28B-unfavorable patients in whom IFNλ4 

mRNA was detected compared with those in IFNλ4-undetected 

patients. However, expressions of suppressive ISGs (RNF125, 

SOCS1, SOCS3 and RNF11) were significantly lower in 

IFNλ4-detected patients. Conclusions: Our data suggest IFNλ4 

have a specific role only in CHC. Intrahepatic expression of 

IFNλ4 is associated with higher expression of antiviral ISGs 

and lower expressions of suppressive ISGs at baseline, resulting 

in the poor responsiveness for IFNα-based therapy for HCV 

infection. 

Disclosures: 


MSD Co., Ltd. 



Squibb 

Mamoru Watanabe - Grant/Research Support: MSD Co., Ltd. 

The following authors have nothing to disclose: Miyako Murakawa, Yasuhiro 

Asahina, Fukiko Kawai-Kitahata, Hiroko Nagata, Syun Kaneko, Sayuri Nitta, 

Takako Watanabe, Yasuhiro Itsui, Mina Nakagawa, Sayuki Iijima, Yujiro Tanaka 

1019 

Lipid-Free apolipoprotein E signaling induces an ABCG1 

driven cholesterol efflux that markedly decreases HCV 

replication 

Emilie Crouchet 1,2 , Mathieu Lefèvre 1,3 , Eloi R. Verrier 1,2 , Thomas 

F. Baumert 1,2 , Catherine Schuster 1,2 ; 1 U1110, Inserm, Strasbourg, 

France; 2 Strasbourg University Hospital, Université de Strasbourg, 

Strasbourg, France; 3 CNRS UPR 9002, Strasbourg, France 

There is an intimate link between the hepatitis C virus (HCV) 

life cycle and its host cell’s lipid metabolism. HCV is associated 

with lipoproteins in blood, forming lipo-viro-particles, which 

are the infectious form of the virus. Apolipoprotein E (apoE), a 

key lipo-viro-particle component, plays an essential role in HCV 

entry, assembly, and egress. Although both the lipoprotein-associated 

and lipid-free forms of apoE can be found in blood, 

the role of lipid-free apoE in the HCV life cycle is unknown. In 

this study, we investigated the effect of lipid-free apoE on the 

HCV life cycle using the HCVcc model system and primary 

human hepatocytes (PHH). A dose-dependent decrease in HCV 

replication was observed when Huh 7.5.1 cells or PHHs were 

treated with increasing amounts of lipid-free apoE. We show 

that lipid-free apoE acts on HCV replication independently of 

previously described apoE receptors, and independently of 

the lipid-free apoE recycling process. By investigating the role 

of lipid-free apoE on actors of hepatic lipid metabolism we 

observed that extracellular lipid-free apoE markedly increases 

cholesterol efflux via the ATP binding cassette sub-family G 

member 1 protein (ABCG1). Our findings highlight a new 

mechanism in lipid metabolism regulation and may provide 

new therapeutic strategies to fight chronic HCV infection.


Disclosures: 

The following authors have nothing to disclose: Emilie Crouchet, Mathieu Lefèvre, 

Eloi R. Verrier, Thomas F. Baumert, Catherine Schuster 

1020 

Substitution in amino acid 70 of hepatitis C virus core 

protein changes the adipokine profile via toll-like receptor 

2/4 signaling 

Masahiko Tameda, Kazushi Sugimoto, Yoshiyuki Takei; Mie University, 

Tsu, Japan 

Background & Aims: It has been suggested that amino acid 

(aa) substitution at position 70 from arginine (70R) to glutamine 

(70Q) in the genotype 1b hepatitis C virus (HCV) core protein 

is associated with insulin resistance and worse prognosis. 

However, the precise mechanism is still unclear. The aim of 

this study was to investigate the impact of the substitution at 

position 70 in HCV core protein on adipokine production by 

murine and human adipocytes. Methods: The influence of treatment 

with HCV core protein (70R or 70Q) on adipokine production 

by both 3T3-L1 and human adipocytes were examined 

with real-time PCR and enzyme-linked immunosorbent assay 

(ELISA), and triglyceride content was also analyzed. The effects 

of toll-like receptor (TLR)2/4 inhibition on IL-6 production by 

3T3-L1 induced by HCV core protein were examined. Results: 

IL-6 production was significantly increased and adiponectin 

production was reduced without a change in triglyceride content 

by treatment with 70Q compared to 70R core protein in 

both murine and human adipocytes. IL-6 induction of 3T3-L1 

cells treated by 70Q HCV core protein was significantly inhibited 

with anti-TLR2 antibody by 42%, and by TLR4 inhibitor 

by 40%. Conclusions: Our study suggests that extracellular 

HCV core protein with substitution at position 70 enhanced IL-6 

production and reduced adiponectin production from visceral 

adipose tissue, which can cause insulin resistance, hepatic 

steatosis, and ultimately development of HCC. 

Disclosures: 

The following authors have nothing to disclose: Masahiko Tameda, Kazushi 

Sugimoto, Yoshiyuki Takei 

1021 

Variability of the hepatitis delta virus quasispecies in 

chronic infection is high and similar to that of the hepatitis 

C virus quasispecies 

Maria Homs 1,2 , Josep Gregori 3 , Hadi Karimzadeh 4 , Damir G. 

Cehic 3 , Maria Blasi 1,2 , Rosario Casillas 3 , David Tabernero 1,2 , 

Josep Quer 1,3 , Michael Roggendorf 4 , Mar Riveiro-Barciela 5 , Rafael 

Esteban 1,5 , Francisco Rodriguez-Frias 1,2 , Maria Buti 1,5 ; 1 Centro de 

Investigacion Biomedica en Red de Enfermedades Hepaticas y 

Digestivas, CIBERehd, Barcelona, Spain; 2 Microbiology, Hospital 

Vall d’Hebron, Barcelona, Spain; 3 Liver Diseases, Research Institute 

Vall d’Hebron, Barcelona, Spain; 4 Institut für Virologie, Technische 

Universität München, München, Germany; 5 Hepatology, 

Hospital Vall d’Hebron, Barcelona, Spain 

Background Hepatitis D virus (HDV) is a satellite RNA virus with 

a small 1.7-kb genome that uses the hepatitis B virus envelope 

to infect hepatocytes. The complexity of the hepatitis D viral 

population has not been described. Hepatitis C virus (HCV) 

is another RNA virus that circulates in infected individuals as 

a quasispecies; the NS5A region is one of the most complex 

of the non-structural encoding HCV genes. Aim To study the 

complexity of the HDV quasispecies in patients with chronic 

hepatitis D and compare it with that of the HCV quasispecies. 

Samples and methods Fifteen serum samples from 10 

chronic HCV- and 5 chronic HDV-infected treatment-naïve 

patients were analyzed. Samples with similar viremia levels 

were selected (mean HCV-RNA 6.51 log copies/mL, SD 0.57 

and HDV-RNA 6.31 log copies/mL, SD 0.61 p=0.84). HCV- 

RNA was quantified by COBAS-AmpliPrep (Roche) (detection 

limits 1.63-7.83 log IU/mL) and HDV-RNA, by an in-house 

method (detection limits 3.5-8.5 log copies/mL). HCV-RNA 

values were converted to log copies/mL. One essential region 

in each virus was deep-sequenced: the NS5A of HCV (positions 

6299-6735, 436 bp) and the C-terminal region of HDV 

antigen (positions 912-1298, 386 bp). Abundance filters for 

deep-sequencing were set at 0.25%. Quasispecies complexity 

was evaluated by determining the mutation frequency (Mf) 

and nucleotide diversity (Pi). Both parameters take into account 

amplicon length, and represent the number of differences/site 

with respect to the most represented haplotye (Mf) or between 

each pair of genomes (Pi). Results are presented as mean and 

standard deviation (SD). Results A total of 187,606 sequences 

were analyzed. Mf and Pi showed a normal distribution and 

significantly correlated (R=0.985, p


gene. Cellular and viral protein expression was evaluated by 

western blot using antibodies vs. HCV-NS5A, SOD1, SOD2, 

catalase, thioredoxin-1, MAT1, MAT2, PKR, STAT1 and actin. 

Results: SAM treatment decreased HCV-RNA levels 50-70% 

compared to untreated control (24-72h). Total glutathione levels 

increased in both cell lines about 50-60% compared to 

control without SAM (6h post-treatment in replicon cells and 2h 

post-treatment in parental cells). Transcriptional expression of 

SOD1, SOD2 and thioredoxin-1 was increased (0.5, 2.5 and 

2 fold-times respectively compared to control) at different time 

points (24-72h). This effect was not observed for catalase. Interestingly, 

there was no significant change in ROS levels in both 

cell types upon SAM treatment at all times assessed, contrary 

to the observed with PDTC exposition where an average of 

30% reduction was detected (0.5-24h). In other hand, MAT1 

expression was increased (2.5 fold-times at 48h) and MAT2 

was decreased upon SAM exposition (24h) compared with 

untreated cells at both transcriptional and translational level 

in both cell lines. SAM treatment does not modify STAT1 and 

PKR expression compared to untreated cells (24-72h), however 

both protein levels were increased upon IFN-a treatment used 

as a control. Conclusions: A possible mechanism by which 

SAM decreases HCV expression could involve modulating 

antioxidant enzymes systems, biosynthesis of glutathione and 

switching MAT2/MAT1 turnover in Hepatitis C virus expressing 

cells. Proteins PKR and STAT1 were stimulated in the presence 

of IFN-a but not with SAM, suggesting that HCV down-regulation 

mediated by SAM is independent of IFN-a pathway. This 

may have clinical application in terms of understanding the 

pathophysiology of the disease. This work was supported by 

CONACYT-BASICA CB-2010-01155082. 

Disclosures: 

The following authors have nothing to disclose: Sonia A. Lozano-Sepulveda, Eduardo 

Bautista-Osorio, Paula Cordero, Linda E. Muñoz, Ana Maria G. Rivas-Estilla 

1023 

Pre-treatment levels of miR-29b are associated with 

response to treatment and stage of fibrosis in patients 

with chronic hepatitis C [CHC] 

Hossein Sendi 1 , Marjan Mehrab-Mohseni 2 , Mark W. Russo 2 , 

Philippe J. Zamor 2 , Paul A. Schmeltzer 2 , Nury Steuerwald 2 , Judith 

Parsons 2 , Mark G. Clemens 4 , Keith Kaplan 3 , W. Carl Jacobs 3 , 

William A. Ahrens 3 , Herbert L. Bonkovsky 1 ; 1 Gastroenterology, 

Wake Forest Baptist Medical Center, Winston Salem, NC; 2 Liver, 

Carolinas Medical Center, Charlotte, NC; 3 Pathology, Carolinas 

Medical Center, Charlotte, NC; 4 Biology, UNC Charlotte, Charlotte, 

NC 

Both responses to treatment and long-term outcomes of hepatitis 

C virus (HCV) infection are critically affected by host genetic 

factors. We aimed to determine whether pre-treatment levels 

of miR122 or miR-29b play any role in response to treatment 

or stage of fibrosis in patients with CHC. A total of 25 CHC 

patients (HCV genotype 1) were included in this study, among 

whom 11 were treated with IFN plus Ribavirin (with or without 

Telaprevir) and 14 remained treatment naïve. The patients 

were classified according to their virological responses: early 

viral response (EVR): patients whose HCV RNA levels at 12 

weeks were reduced by 2-log 10 

or more compared to baseline. 

Non-EVR (n-EVR): patients whose HCV RNA levels were not 

reduced or reduced less than 2- log 10 

. Using quantitative real 

time PCR, we compared pre-treatment levels of hepatic miR- 

122, and miR-29b (normalized to SNORD44) in the patients 

based on their response to treatment. We found that pre-treatment 

levels of miR-122 were slightly lower in patients with EVR 

than those with n-EVR (49 vs 59, p>0.05). However, pre-treatment 

levels of miR-29b were significantly lower in patients 

with EVR than those with n-EVR (0.015 vs 0.130, p


duced HCV (JFH-1; genotype 2a). IRF9, STAT1, and STAT2 

were overexpressed by lentiviral transduction, or their expression 

was silenced with siRNAs. The expression of the negative 

regulators of type I IFN signaling, USP18 and ISG15, was 

studied in HCV-infected liver and IFN-λ 3 

treated hepatoma 

cells, and their regulation by U-ISGF3 was analyzed. Results 

The level of U-ISGF3, but not tyrosine phosphorylated STAT1, 

is significantly elevated in response to IFN-λ and IFN-β during 

chronic HCV infection. U-ISGF3 prolongs the expression of a 

subset of ISGs and restricts HCV chronic replication. However, 

paradoxically, high levels of U-ISGF3 also conferred unresponsiveness 

to IFN-α therapy. As a mechanism of U-ISGF3-induced 

resistance to IFN-α, we found that ISG15, a U-ISGF3-induced 

protein, sustains the abundance of USP18. Silencing ISG15 

decreased the level of USP18 in IFN-λ 3 

treated hepatoma cells 

and restored USP18-mediated attenuation of STATs phosphorylation 

after treatment of IFN-α. Conclusions Our data demonstrate 

that U-ISGF3 induced by IFN-λs and -β drives prolonged 

expression of a set of ISGs, leading to chronic activation of 

innate responses and conferring a lack of response to IFN-α in 

HCV-infected liver. 

Disclosures: 

The following authors have nothing to disclose: Pil Soo Sung, HyeonJoo Cheon, 

Do Youn Park, Hyung-Il Seo, Su-Hyung Park, Seung Kew Yoon, George R Stark, 

Eui-Cheol Shin 

1026 

The expression of Immune-miRs could contribute to the 

immunopathogenesis of HCV and be modified by 1(OH) 

vitamin D3 supplementation 

Yasuteru Kondo 1 , Tatsuki Morosawa 1 , Masashi Ninomiya 1 , 

Yasuyuki Fujisaka 1 , Yasuhito Tanaka 2 , Takayuki Kogure 1 , Jun 

Inoue 1 , Teruyuki Umetsu 1 , Tooru Shimosegawa 1 ; 1 Gastroenterology, 

Tohoku University Hospital, Sendai, Japan; 2 Nagoya City 

University, Nagoya, Japan 

[Background] We reported that the expression profiles of serum 

miRNAs could be important biomarkers for the diagnosis of 

various liver diseases (PLoS One 2013). It has been reported 

that various kinds of miRNAs known as immune-miRs could contribute 

to the immune regulation. [Aim] The aim of this study is 

to analyze the biological significance of immune-miRs expression 

on specific immune cells in chronic hepatitis C (CH-C) 

patients. [Material and Methods] Patients: Permission for the 

study was obtained from the ethics committee at our institute 

(2013-1-268). Twenty CH-C patients, 20 chronic hepatitis B 

patients (CH-B), and 10 healthy subjects were examined by 

deep sequencing analysis (Illumina G2X). Moreover, 50 CH-C 

patients (genotype 1b and high viral load) and control patients 

(CH-B, NASH, and healthy subjects) were examined by validation 

analysis. Deep sequencing analysis: Illumina deep 

sequencing for the initial screening to determine the read numbers 

of miRNAs expression in serum and PBMCs was carried 

out. Immune cells isolation: CD3 + T cells, CD14 + monocytes, 

CD19 + B cells and CD56 + NK cells were isolated using the 

magnetic beads method. Transfection of HCV individual protein 

expressing plasmids: The HCV individual expression plasmids 

(HCV-E1, E2, Core, NS3, NS4 and NS5A) were transfected 

into immune cells to identify the HCV proteins responsible for the 

suppression of miR146b-5p. Silencing and forced expression of 

miR146b-5p: Transfections of miRNA mimic (#MC10105) and 

miRNA inhibitor into THP-1, Jurkat, Molt-4, Raji and human 

primary lymphoid cells using 4D-nucleofector TM were carried 

out. [Results] The expressions of five miRNAs (miR146b-5p, 

miR-181a-2-3p, miR-204-5p, miR374a-3p and miR374a-5p) 

in the serum were significantly lower than those of the control 

groups. Among them, miR146b-5p was abundantly expressed 

in PBMCs. The expression of miR146b-5p in CD3 + T cells and 

CD14 + monocytes in CH-C patients was significantly lower 

than those in control groups. The silencing of miR146b-5p in 

the monocytes could significantly enhance the expression of 

CXCL10, TGF-β and IL10 under the stimulation (p


a treatment history with PEG-IFN/RBV (10 were prior null 

responders and 11 were prior relapsers). The SVR rate was 

86 % (6/7 patients) in treatment naïve patients, 82 % (9/11 

patients) in prior relapsers, and 40 % (4/10 patients) in prior 

null responders. the NS3 and the NS5A RAVs at baseline were 

detected in 71% (20/28 patients) and 81% (17/21 patients) 

by ultra-deep sequencing. No difference was found in the 

number of the patients with the NS3 RAVs before treatment 

between SVRs and non-SVRs (85% [11/19] vs. 71% [5/9], 

p=0.91). However, at the early phase of the treatment, increasing 

prevalence of the RAVs was more frequently found in non- 

SVRs more frequently than in SVRs (78 % [7/9], 32 % [6/19], 

p2 and p2 and an unadjusted p-value of



suggesting that HCV-ICs persist in an even higher proportion 

of patients. Studies are ongoing to assess the clinical implications 

of persistent HCV-specific ICs. CONCLUSIONS. We 

demonstrated for the first time that HCV-ICs persist in a majority 

of patients who had spontaneously cleared HCV or achieved 

SVR months to years earlier. Evidence of ongoing presence of 

HCV proteins after HCV clearance extends our understanding 

of pathogenesis. When using HCV-Ag assays, serum sample 

denaturation may result in failure to differentiate active from 

resolved HCV infection. 

Disclosures: 

The following authors have nothing to disclose: Ke-Qin Hu, Wei Cui 

1032 

Very low prevalence of Hepatitis C in HIV negative 

men who have sex with men in Tel Aviv: A prospective 

cohort analysis 

Ella Veitsman 1,7 , Adir Yanko 2 , Yuval Livnat 3,4 , Margalit Lorber 5,6 , 

Ziv Ben Ari 1,7 ; 1 center for liver diseases, sheba medical center, 

Ramat Gan, Israel; 2 Israel LGBT organization, Levinsky clinic, Tel 

Aviv, Israel; 3 Israel AIDS Task force, Tel Aviv, Israel; 4 Tel Aviv University, 

Tel Aviv, Israel; 5 Immunology Department, Rambam Health 

Care Campus, Haifa, Israel; 6 The Ruth and Bruce Rappaport 

School of medicine, Israel institute of Technology, Haifa, Israel; 

7 Sackler School of medicine, Tel Aviv University, Tel Aviv, Israel 

Background: In light of highly effective existing and upcoming 

antiviral treatments, it is necessary to define additional risk 

groups of Hepatitis C virus (HCV) transmission. The occurrence 

of sexual transmission of HCV in human immunodeficiency 

virus (HIV) positive men who have sex with men (MSM) is well 

described and on the rise all over the world. Outbreaks of hepatitis 

C in this population have been linked to several risk factors 

including unprotected anal sex, use of recreational drugs, 

and concomitant sexually transmitted infections. This has led 

some experts to suggest that HIV negative MSM also have a 

high rate of hepatitis C but are simply not getting tested for it. 

However epidemiological studies on possible sexual transmission 

of hepatitis C in HIV negative MSM have shown significantly 

variable results in different countries. Aim: To determine 

the prevalence of positivity for antibodies to hepatitis C virus 

(anti-HCV) and the potential for sexual transmission of the 

virus in HIV negative MSM. Patients and methods: Participants 

were prospectively recruited from a day-MSM clinic in Tel Aviv 

during the period from July 2014 to April 2015. Participants 

self-completed a short questionnaire providing demographic 

and behavioral data. They also donated a sample of blood 

that was subsequently tested for antibodies to HCV and other 

selected pathogens (HIV, syphilis, gonorrhea and chlamydia). 

Results: In total 730 MSM HIV negative participants, median 

age 28.0 (range 16-64) completed the questionnaire and 

underwent HCV testing. The majority of the tested population 

was born in Israel. 85 participants (11.87%) were immigrants 

from other countries. Overall three of 730 tested were found 

sero-positive for antibodies to HCV. All three were born in 

Israel. The overall prevalence was 0.41% (95% CI: 0.21- 

0.8%). Further HCV RNA testing was negative in all three anti 

HCV positive men. Therefore no one was found harboring 

a replicating virus. Also no HCV sero-conversion was found 

during the study period. None of the three anti HCV positive 

participants reported known risk factors for HCV contraction 

except for unprotected sex in one of them. Conclusion: Our 

findings suggest that in Israel Hepatitis C among MSM without 

diagnosed HIV is much less prevalent than in the general 

population. There is no evidence of elevated rates of sexual 

transmission of HCV among MSM without HIV-infection. 

Disclosures: 

Yuval Livnat - Grant/Research Support: Abbvie, Neopharm Israel, Janssen 

The following authors have nothing to disclose: Ella Veitsman, Adir Yanko, Margalit 

Lorber, Ziv Ben Ari 

1033 

The Antibody response to hepatitis B virus vaccination 

is not influenced by the hepatitis C virus viral load in 

patients with chronic hepatitis C 

Roseane P. Medeiros, Marta Lopes, Daniel F. Mazo, Claudia P. 

Oliveira, Patricia M. Zitteli, João Renato R. Pinho, Flair J. Carrilho, 

Mario G. Pessoa; Gastroenterology, University of Sao Paulo 

School of Medicine, Sao Paulo, Brazil 

Background: Some immunogenicity studies of anti-HBV vaccine 

in patients with chronic HCV infection have demonstrated a 

diminished response ranging from 63.6% to 72.9% on seroconversion 

rate, compared to 90.9% to 93.9% in healthy controls. 

One possible explanation is the high HCV viral load in 

some patients. Aims: To evaluate the impact of HCV viral load 

on anti-HBV vaccination response in treatment naive chronic 

HCV patients without cirrhosis. Methods: 110 chronic HCV 

adult patients without cirrhosis were randomized to receive 

anti-HBV vaccination regimen at standard 3 doses (0, 1 and 

6 months) of 20ug, or higher dose of 40ug. Response to vaccination 

was measured by titers of anti-HBs 1 and 6 month 

after the last dose of anti-HBV vaccine. Healthy controls were 

negative to anti-HCV, anti-HBc, HBsAg and anti-HBs antibodies, 

and received standard 3 doses of 20ug at intervals of 0, 

1 and 6 months, and were also evaluated for anti-HBs titers. 

Results: Of the 110 HCV vaccinated patients, the seroconversion 

rate (anti-HBs ≥ 10IU/mL) was 74.5% (82/110). Out of 

the 45 healthy controls vaccinated with standard dose, we 

observed a seroconversion rate of 93.3% (42/45). Variables 

included in the logistic regression model were: Age, Gender, 

HCVRNA and Liver Fibrosis by Metavir (F0/F1 versus F2/F3). 

Age was the only variable that negatively influenced anti-HBs 

titers (p=0.003) Conclusions: Our study had demonstrated that 

chronic HCV patients without cirrhosis presented impaired anti- 

HBV vaccine response compared to healthy controls, similar to 

data previously demonstrated in the literature. This impairment 

is apparently not influenced by HCV viral load. 

Disclosures: 

João Renato R. Pinho - Employment: Hospital Israelita Albert Einstein 

The following authors have nothing to disclose: Roseane P. Medeiros, Marta 

Lopes, Daniel F. Mazo, Claudia P. Oliveira, Patricia M. Zitteli, Flair J. Carrilho, 

Mario G. Pessoa


1034 

Approved All-oral Sofosbuvir Regimens are Safe and 

Highly Effective in Patients with Hereditary Bleeding 

Disorders 

Christopher Walsh 1 , Kimberly Workowski 2 , Norah Terrault 3 , Paul 

Sax 4 , Alice Cohen 5 , Christopher L. Bowlus 6 , Arthur Y. Kim 7 , Robert 

H. Hyland 8 , Jing Wang 8 , Luisa M. Stamm 8 , Diana M. Brainard 8 , 

John G. McHutchison 8 , Annette Von Drygalski 9 , Frank S. Rhame 10 , 

Michael W. Fried 11 , Peter Kouides 12 , Gayle Balba 13 , K. Rajender 

Reddy 14 ; 1 Mount Sinai Hospital, New York, NY; 2 Emory University, 

Atlanta, GA; 3 University of California, San Francisco, San 

Francisco, CA; 4 Brigham and Women’s Hospital, Boston, MA; 

5 Newark Beth Israel Center - Barnabas Health, Newark, NJ; 6 University 

of California, Davis, Sacremento, CA; 7 Massachusetts General 

Hospital, Boston, MA; 8 Gilead Sciences, Inc, Foster City, CA; 

9 University of California, San Diego, San Diego, CA; 10 Abbott 

NW Infectious Disease Clinic, Minneapolis, MN; 11 University of 

North Carolina at Chapel Hill, Chapel Hill, NC; 12 The Mary M 

Gooley Hemophilia Center, Rochester, NY; 13 Georgetown University 

Hospital, Washington, DC; 14 University of Pennsylvania, 


Background: Patients with bleeding disorders have high rates 

of Hepatitis C Virus (HCV) infection due to exposure to contaminated 

blood products prior to 1992. They have historically 

been excluded from HCV clinical trials due to concern for 

unique adverse events, comorbidities, and/or their underlying 

diagnosis. This study evaluated the safety and efficacy of ledipasvir/sofosbuvir 

(LDV/SOF, genotype [GT] 1 and 4 infection) 

and SOF+RBV (GT 2 and 3 infection) in patients with bleeding 

disorders with or without HIV coinfection. Methods: GT1 or 

4 HCV-infected patients received LDV/SOF (90mg/400mg) 

fixed dose combination daily for 12 weeks; treatment was 

extended to 24 weeks for those who were treatment experienced 

with cirrhosis following FDA approval of LDV/SOF. 

GT 2 or 3 HCV-infected patients received SOF 400 mg daily 

+RBV (1000 or 1200 mg/day divided BID) for 12 weeks or 

24 weeks, respectively. Results: 120 patients were enrolled at 

13 sites in the USA. The majority were male (94%), Caucasian 

(76%), and IL28B non-CC genotype (69%). 103 patients were 

infected with GT1 HCV infection, 10 with GT2, 6 with GT3, 

and 1 with GT4. The most common bleeding disorders were 

Hemophilia A (65%) and Hemophilia B (26%). Patients with 

Type 1 or 3 Von Willebrand’s Disease (3% each), Type 2 Von 

Willebrand’s Disease (2%), and Factor XI deficiency (


similar between groups. During treatment, patients receiving 

FDC showed improvement of PRO scores (+6.0% in activity/ 

energy (AE) of CLDQ-HCV, +5.0% in fatigue score (FS) of FAC- 

IT-F, +6.8% in physical component of SF-36; all p


nosed between 1998-2014) with SCD in remission (with sickle 

cell history > 30 years) All patients were placed LDV 90 mg 

+ SOF 400 mg a day; with food for 12 weeks Patient characteristics: 

the table will be included in the oral presentation 

(slide) or poster Medications allowed: Hydroxyurea, Tylenol, 

MS Contin, Methadone, Antibiotics (Levofloxacin, Doxycycline, 

Colchicine, Metronidazole), epoetin alpha Exclusion: Decompensated 

cirrhotics, HIV, HBV, Sepsis, brittle SCD with frequent 

flares, uncontrolled DM, cardiomyopathy, CHF NYHA class 

III-IV, CKD, chronic osteomyelitis, active IVDU, Daily alcohol > 

30 grams, Deferoxamine for 12 weeks. Side events: Nausea- 

9/24, constipation 6/24, diarrhea 2/24, headache 6/24, 

abdominal pain 3/24, renal colic 1/24, insomnia 8/24, anemia 

2/24, hyperbilirubinemia 4/24, UTI 3/24 Conclusion: 

This study demonstrates that LDV and SOF combination in SCD 

patients with CHC is safe and well tolerated; with an SVR12 of 

91.67% (22/24) with 8.3% (2/24) viral failure (in concomitant 

genotypes; 1a/4c and 1a/3c). 

Results 

(range 2-8 weeks) and remained negative thereafter. One 

patient who was RBV-ineligible and received SOF/LDV without 

RBV for only 12 weeks due to insurance issues developed virological 

relapse between 4 and 8 weeks after treatment discontinuation. 

Conclusions: Treatment with SOF/LDV with or without 

RBV for 12-24 weeks was very well tolerated and resulted in 

an excellent on-treatment virological response in HCV GT1 

relapsers to SMV+SOF treatment. SVR12 data will be reported 

when available. 

On-Treatment Virological Response Rates 

Disclosures: 

Surakit Pungpapong - Grant/Research Support: BMS, Gilead 

The following authors have nothing to disclose: Michael D. Leise, Kymberly D. 

Watt, Hugo E. Vargas, Andrew P. Keaveny, Bashar A. Aqel 

Disclosures: 

The following authors have nothing to disclose: Patrick Basu, Niraj J. Shah, Nimy 

John, M. Aloysius, Robert Brown 

1038 

Multicenter Experience using Sofosbuvir/Ledipasvir with 

or without Ribavirin to Treat Hepatitis C Genotype 1 

Relapsers after Simeprevir and Sofosbuvir Treatment 

Surakit Pungpapong 1,2 , Michael D. Leise 3 , Kymberly D. Watt 3 , 

Hugo E. Vargas 4 , Andrew P. Keaveny 1,2 , Bashar A. Aqel 4 ; 

1 Transplant, Mayo Clinic, Jacksonville, FL; 2 Gastroenterology & 

Hepatology, Mayo Clinic, Jacksonville, FL; 3 Gastroenterology & 

Hepatology, Mayo Clinic, Rochester, MN; 4 Gastroenterology & 

Hepatology, Mayo Clinic, Scottsdale, AZ 

Background: Approximately 10-15% of patients with hepatitis 

C genotype 1 (HCV GT1) experience virological relapse after 

all-oral antiviral regimen using simeprevir (SMV) and sofosbuvir 

(SOF). The efficacy and safety of treating such relapsers using 

sofosbuvir/ledipasvir fixed-dose combination (SOF/LDV) with/ 

without ribavirin (RBV) has not been described to date. Objective: 

To report the virological response and safety of SOF/ 

LDV with or without RBV for 12-24 weeks in treating HCV 

GT1 relapsers after SMV+SOF treatment. Results: To date, 42 

patients (26% post-LT, 79% male, 17% non-white, 80% subtype 

1a, 86% IL28B CT/TT, 76% F3-4) started SOF/LDV with 

or without RBV at a median of 20 weeks (range 7-55 weeks) 

after the last dose of SMV+SOF treatment. Five and 25 patients 

started SOF/LDV with RBV (dose adjusted for renal function) for 

12 and 24 weeks, respectively; while 12 patients who were 

RBV-ineligible received SOF/LDV without RBV for 24 weeks. 

Minimal adverse events were reported in those without RBV; 

48% patients who received RBV developed significant anemia 

requiring RBV dose reduction and/or discontinuation. In 

LT recipients, minimal immunosuppression dose adjustments 

were required and no biopsy-proven acute rejection occurred. 

On-treatment virological response rates are shown in the Table. 

Of 39 patients who received treatment at least 4 weeks, all 

achieved undetectable HCV RNA at a median of 4 weeks 

1039 

RUBY-I: Ombitasvir/Paritaprevir/Ritonavir + Dasabuvir 

+/- Ribavirin in Non-cirrhotic HCV Genotype 1-infected 

Patients With Severe Renal Impairment or End-Stage 

Renal Disease 

Paul J. Pockros 1 , K. Rajender Reddy 2 , Parvez S. Mantry 3 , Eric 

Cohen 4 , Michael Bennett 5 , Mark S. Sulkowski 6 , David Bernstein 7 , 

Thomas Podsadecki 4 , Daniel E. Cohen 4 , Nancy Shulman 4 , Deli 

Wang 4 , Amit Khatri 4 , Manal Abunimeh 4 , Eric Lawitz 8 ; 1 Scripps 

Clinic, La Jolla, CA; 2 University of Pennsylvania, Philadelphia, 

PA; 3 The Liver Institute at Methodist Dallas, Dallas, TX; 4 AbbVie 

Inc., North Chicago, IL; 5 Medical Associates Research Group, 

San Diego, CA; 6 Johns Hopkins University, Baltimore, MD; 7 North 

Shore University Hospital, Manhasset, NY; 8 The Texas Liver Institute, 

University of Texas Health Science Center, San Antonio, TX 

Background: HCV is common among patients (pts) with endstage 

renal disease. Co-formulated ombitasvir/paritaprevir/r 

(paritaprevir identified by AbbVie and Enanta, co-dosed with 

ritonavir [r]) and dasabuvir (3D) do not require dose adjustment 

in pts with renal insufficiency. In Phase 3 trials, 3D+/- RBV 

showed high SVR rates and low rates of discontinuation due to 

adverse events in HCV genotype 1 (GT1)-infected pts. RUBY-I is 

an ongoing open-label study evaluating 3D+/-RBV in pts with 

stage 4 or 5 chronic kidney disease (CKD) and GT1 infection. 

Methods: Cohort 1 of RUBY-I enrolled treatment-naïve, non-cirrhotic 

adults with GT1 infection and CKD stage 4 (estimated 

GFR 15-30 mL/min/1.73m 2 ) or 5 (eGFR


pts with stage 4 or 5 CKD in this ongoing trial, 3D+/-RBV has 

been well tolerated, with no premature treatment discontinuations. 

Hemoglobin decreases were managed with RBV interruption, 

which does not appear to affect efficacy. There have been 

no virologic failures to date. 

Cohort 1: Baseline characteristics. 

Disclosures: 












Salix 



BMS 




Gilead 

Thomas Podsadecki - Employment: AbbVie; Stock Shareholder: AbbVie 

Daniel E. Cohen - Employment: AbbVie; Stock Shareholder: AbbVie 


Deli Wang - Employment: AbbVie Inc 

Amit Khatri - Employment: AbbVie, Inc; Patent Held/Filed: AbbVie, Inc; Stock 

Shareholder: AbbVie, Inc 

Manal Abunimeh - Employment: AbbVie 








The following authors have nothing to disclose: Eric Cohen, Michael Bennett 

1040 

Impact of baseline albumin levels in a Phase 3 study 

(OPTIMIST-2) of 12 weeks of simeprevir (SMV) plus 

sofosbuvir (SOF) in treatment-naïve or -experienced 

patients with chronic HCV genotype 1 infection and cirrhosis 

Eric Lawitz 1 , Eric M. Yoshida 2 , Reem H. Ghalib 3 , Marcelo Kugelmas 

4 , Ronald Kalmeijer 5 , Katrien Janssen 6 , Oliver Lenz 6 , Bart 

Fevery 6 , Guy De La Rosa 7 , Rekha Sinha 5 , James Witek 5 ; 1 Texas 

Liver Institute, University of Texas Health Science Center, San Antonio, 

TX; 2 University of British Columbia, Vancouver, BC, Canada; 

3 Texas Clinical Research Institute, Arlington, TX; 4 South Denver 

Gastroenterology, PC, Denver, CO; 5 Janssen Research & Development, 

LLC, Titusville, NJ; 6 Janssen Infectious Diseases BVBA, 

Beerse, Belgium; 7 Janssen Global Services, LLC, Titusville, NJ 

Purpose: In the Phase 3 OPTIMIST-2 (NCT02114151) study 

in HCV GT1-infected pts with cirrhosis, SMV+SOF for 12wks 

achieved superiority in sustained virologic response 12wks 

after end of treatment (SVR12) vs a historical control. This analysis 

evaluated SVR12 by baseline (BL) albumin. Methods: Pts 

with documented cirrhosis received 12wks of SMV 150mg 

QD+SOF 400mg QD. Primary endpoint: SVR12. Univariate 

logistic regression established the relationship between BL albumin 

and SVR12. SVR12 rates were evaluated post hoc by 

BL albumin and presence of other factors: BL Q80K polymorphism; 

IL28B genotype; BMI; platelet count; prior treatment; 

sex. Results are presented for the non-VR excluded population. 

Results: 100pts were treated (male 81%; median age 58yrs; 

Black/African American 18%; IL28B non-CC 72%; GT1a 69% 

[of these: +/- Q80K 46/54%]; treatment-naïve 49%; BL albumin, 

median 39 [range 29–47] g/L). Overall SVR12 was 85%. 

A strong univariate relationship was observed between BL albumin 

and SVR12 (AUC ROC 0.79). Analysis of the correlation 

of BL albumin and SVR12 rates identified a level of 40g/L as 

an optimal cut-off. 51% and 49% pts had BL albumin


Disclosures: 

















Marcelo Kugelmas - Advisory Committees or Review Panels: Merck, Abbvie, 

Janssen, Salix; Consulting: Abbvie, Merck, Gilead, Janssen; Grant/Research 

Support: Abbvie, Intercept, Hologic, Gilead, Janssen, Roche, Anadys, Salix; 

Speaking and Teaching: Abbvie, Gilead, Merck, Janssen, Salix 

Ronald Kalmeijer - Employment: Johnson and Johnson 

Katrien Janssen - Employment: Janssen Pharmaceutica NV 

Oliver Lenz - Employment: Janssen; Stock Shareholder: Janssen (J&J) 

Bart Fevery - Employment: Janssen Infectious Diseases BVBA; Stock Shareholder: 

Janssen Infectious Diseases BVBA 

Guy De La Rosa - Employment: Johnson & Johnson 

Rekha Sinha - Employment: Janssen Pharmaceutica 

James Witek - Employment: Johnson & Johnson; Stock Shareholder: Johnson & 

Johnson 

1041 

Twelve Weeks of Sofosbuvir plus Ribavirin is Effective 

for Treatment of Genotype 2 HCV in Difficult to Treat 

U.S. Veterans with Cirrhosis – Results of the VAL- 

OR-HCV Study 

Samuel B. Ho 1 , Alexander Monto 2 , Adam Peyton 3 , David E. 

Kaplan 4 , Sean Byrne 5 , Scott Moon 5 , Yanni Zhu 5 , Star Seyedkazemi 

5 , Lorenzo Rossaro 5 , Diana M. Brainard 5 , William Guyer 5 , 

Obaid S. Shaikh 6 , Michael Fuchs 7 , Timothy R. Morgan 8 ; 1 Gastroenterology, 

VA San Diego Healthcare System, San Diego, CA; 

2 Gastroenterology, UCSF/SFVAMC, San Francisco, CA; 3 Gastroenterology, 

Miami VA Healthcare System, Miami, FL; 4 Gastroenterology, 

Philadelphia VA Medical Center, Philadelphia, PA; 5 Gilead 

Sciences, Foster CIty, CA; 6 VA Pittsburgh Healthcare System, Pittsburgh, 

PA; 7 Richmond VA Medical Center, Richmond, VA; 8 Gastroenterology, 

VA Long Beach Healthcare System, Long Beach, CA 

Background: The prevalence of chronic HCV in U.S. veterans 

is approximately 4%. Veterans often have medical or psychiatric 

comorbidities that limit their enrollment in registration trials. 

Registries and surveillance reports show lower efficacy 

and higher discontinuation rates among veterans than in other 

patients. Data are limited on use of sofosbuvir (SOF) and ribavirin 

(RBV) for treatment of patients with cirrhosis from genotype 

2 (GT2) HCV infection, due to relatively small number 

of such subjects enrolled in clinical trials. Therefore, we conducted 

an open-label study to evaluate the safety and efficacy 

of 12 weeks of SOF+RBV in GT2 HCV infected veterans with 

cirrhosis. Methods: VALOR-HCV was conducted exclusively at 

15 U.S. VA sites in both treatment-naïve (TN) and treatment-experienced 

(TE) GT2 patients with compensated cirrhosis. The 

study had expanded inclusion criteria and did not exclude 

patients with active alcohol and polysubstance abuse. Subjects 

received 12 weeks of oral SOF 400 mg daily + weight-based 

RBV 1000-1200 mg/day in divided doses. The primary endpoint 

was sustained virologic response at 12 weeks after end 

of treatment (SVR12). Results: 66 cirrhotic GT2 HCV infected 

subjects, 47 TN and 19 TE, were enrolled. All subjects were 

male, mean age was 63 years (range: 49 - 85 years) and 

mean BMI was 30.2 kg/m 2 (range: 17.7-51.0 kg/m 2 ). Fifteen 

percent of subjects were African American, 9% Hispanic, 

and 33% classified as intolerant to or ineligible for treatment 

with interferon. Among subject comorbidities were vascular 

(71.2%), metabolic (62.1%), and psychiatric (56.1%) disorders. 

Cirrhosis was diagnosed by FibroTest (58%), FibroScan 

(26%), biopsy (12%), and imaging (5%). Ten TN (21%) and 2 

TE (11%) subjects had SOF adherence rates of


respondents ascertained prior awareness of HCV infection status. 

Population percentages and p-values were weighted to 

adjust for sampling design. Results: 162 of 458 NHANES 

respondents who tested positive for HCV antibodies or RNA 

during 2003-2012 participated in the follow-up survey. Of 

these, 91 (59%) reported that they were already aware of 

their HCV status when informed of their positive HCV test in 

NHANES. Rates of prior HCV status awareness varied considerably; 

71% of adults born in 1945-1965 reported prior 

awareness, for example, compared to 28% of adults not in 

major risk groups (p=0.096). People living above 2x Federal 

poverty level (FPL) were more likely (72%) than those below 2x 

FPL (48%) to have been aware of their HCV status (p=0.019). 

Conclusions: HCV screening practices before 2010 appear to 

have identified just over half of total HCV cases in the non-institutionalized 

US population. Furthermore, large disparities exist 

in awareness of HCV status, particularly among marginalized 

and seemingly low-risk groups. With new therapies promising 

higher certainty of cure for people with HCV, development of 

effective screening policies is of great importance. 

Disclosures: 

Mark T. Linthicum - Employment: Precision Health Economics 

Timothy R. Juday - Employment: AbbVie; Stock Shareholder: AbbVie 

Gigi Moreno - Consulting: AbbVie 

Yuri Sanchez - Employment: AbbVie; Stock Shareholder: AbbVie 

Darius Lakdawalla - Consulting: AbbVie Pharmaceuticals; Stock Shareholder: 

Precision Health Economics 

Steven Marx - Employment: AbbVie; Stock Shareholder: AbbVie 

The following authors have nothing to disclose: Brian R. Edlin 

1043 

Sustained Virologic Response to Direct Acting Antiviral 

Therapy for Chronic Hepatitis C Infection is Associated 

with Improvement of Components of the Metabolic Syndrome 

Mark Pedersen 1 , David W. Backstedt 2 , Bobby Kakati 2 , Myunghan 

Choi 3 , Anil B. Seetharam 4,5 ; 1 Department of Internal Medicine, 

Banner University Medical Center, Tempe, AZ; 2 Department of 

Gastroenterology, Banner University Medical Center, Phoenix, AZ; 

3 Arizona State University College of Nursing and Health Care 

Innovation, Tempe, AZ; 4 Banner Transplant and Advanced Liver 

Disease, Phoenix, AZ; 5 University of Arizona College of Medicine 

Phoenix, Phoenix, AZ 

Rationale: Chronic hepatitis C virus (HCV) infection has been 

linked to the metabolic syndrome. While direct acting antiviral 

(DAA) therapy for HCV is often successful in achieving 

sustained virologic response (SVR), pleiotropic effects on 

components of the metabolic syndrome have not been fully 

characterized. Aim: To evaluate the effect of DAA HCV therapy 

on components of the metabolic syndrome. Methods: A 

prospective, cohort study of consecutively enrolled, mono-infected 

chronic HCV patients initiated on treatment with a DAA 

regimen. Consecutively enrolled patients received one of the 

following: 1) pegylated interferon alfa 2a (IFN) + sofosbuvir 

(SOF) + ribavirin (RBV); 2) SOF + RBV; 3) SOF + simeprevir 

(SMV) or 4) ledipasvir (LDV) + SOF in accordance with AASLD 

guidelines. Metabolic parameters including: body mass index 

(BMI), systolic blood pressure (BP), diastolic BP, fasting glucose, 

total cholesterol, high density lipoprotein (HDL), low density 

lipoprotein (LDL), triglycerides, and fasting glucose were measured 

prior to and at the end of treatment. Subgroup analysis 

evaluated metabolic profile changes between groups achieving 

SVR at 12 weeks. Changes in metabolic parameters were 

analyzed with student’s t test, p < 0.05 significant. Results: 

Patients (n=218) completed treatment with a DAA based regimen: 

IFN + SOF + RBV (17%); SOF + RBV (54.6%); SMV + 

SOF (23.4%) and SOF + LDV (6%). Of 218 enrolled, 61.5% 

genotype 1 and 56.4% cirrhotic. Previous IFN exposure: 64% 

naïve followed by non-response (26%) and relapse (10%). The 

highest prevalence of concomitant metabolic syndrome parameter 

prior to treatment was hypertension (38.5%) followed by 

diabetes (16.5%) and hyperlipidemia (15.3%). For the entire 

cohort, end of treatment (ETR) was 98%, with 79% going on 

to achieve SVR 12 (19% relapsers). At ETR, significant reductions 

in BMI, systolic and diastolic BP, total cholesterol, and 

LDL were noted in the entire cohort (all p


analyzed with MEGA 5.0 software to identify RAVs to nine 

DAAs (Boceprevir, Telaprevir, Simeprevir, Paritaprevir, Daclatasvir, 

Ledipasvir, Omitasvir, Sofosbuvir and Dasabuvir) in the 

HCV genome, which were summarized from the up-to-date 

literature available. Results: This analysis collected 1459 fulllength 

HCV sequences from GenBank, 70.7% of which carried 

at least one dominant resistance variant. Geographically, the 

highest RAV frequency occurred in Asia (87.7%), followed 

by Africa (73.8%), Europe (69.0%) and America (63.3%). 

The highest RAV frequency was observed in genotype (GT) 6 

sequences (83.8%), followed by GT2 (83.2%), GT4 (77.6%), 

GT1 (67.6%) and GT3 (50.0%). Furthermore, 46.0%, 30.6% 

and 10.8% of sequences were observed RAVs to NS5A inhibitors, 

NS3 protease inhibitors and their combinations respectively. 

However, RAVs to NS5B nucleos(t)ide inhibitor (NI) 

and NI-based combinations were uncommon observed (< 3% 

of sequences). As expected, RAVs were rare (0.2% to 2.1%) 

detected to the IFN-free regimens recommended by the currently 

guideline. Conclusion: The global overall prevalence of 

DAA-induced RAVs was high without regard for geography or 

genotype. Similarly, RAVs to NS5A and NS3 inhibitors were 

mainly observed. In contrast, NS5B NI-based multi-DAA regimens 

had a low prevalence of RAVs, suggesting that NS5B 

NI-based DAA regimens are the most promising strategies for 

a long-term HCV infection cure. This result was consistent with 

the currently recommended IFN-free regimens. 

Disclosures: 

The following authors have nothing to disclose: Peng Hu, Zhi-wei Chen, Hu Li, 

Hong Ren 

SOLAR1 (NCT01938430) and SOLAR 2 (NCT02010255) 

studies. On treatment and post-treatment HCV RNA measurements 

from LT patients who received CyA or TAC were 

compared. Patients who took both or neither of the immunosuppresants 

were excluded. Final data for post-treatment Week 

12 for SOLAR 2 are pending and will be presented; however, 

relapse rates based on post-treatment Week 4 data were calculated. 

HCV RNA was measured using the Roche COBAS® 

Ampliprep®/Cobas TaqMan HCV Test, Version 2.0 (CAP/ 

CTM HCV v2.0) with a lower limit of quantification (LLOQ) of 

15 IU/mL. Results: A total of 405 post-transplantation patients 

were included in the analysis: 309 (76%) patients had received 

TACand 96 (24%) had received CyA. The number of patients 

with decompensated disease receiving TAC and CyA was 73 

(24%) and 22 (23%) respectively. Median, Q1, and Q3 baseline 

HCV RNA values were similar among patients receiving 

TAC (6.5, 6.0, 6.8) and CyA (6.6, 6.1, 6.9). The percentage 

of patients with an HCV RNA < 800,000 IU/mL was 21% 

versus 18%, for TAC and CyA. The percent of patients who 

were







Gilead 














1046 

Effectiveness of 8 or 12 week LDV/SOF in treatment-naïve 

patients with non-cirrhotic, genotype 1 Hepatitis 

C: Real-world experience from the TRIO Network 

Michael P. Curry 1 , Bruce Bacon 2 , Steven L. Flamm 3 , Naoky CS C. 

Tsai 4 , Douglas Dieterich 5 , Scott Milligan 6 , Kris V. Kowdley 7 ; 1 Beth 

Israel Deaconess Medical Center, Boston, MA; 2 Saint Louis University 

School of Medicine, St. Louis, MO; 3 Northwestern University 

Feinberg School of Medicine, Chicago, IL; 4 Queens Medical Center, 

University of Hawaii, Honolulu, HI; 5 Icahn School of Medicine 

at Mount Sinai, New York, NY; 6 Trio Health Analytics, Newton, 

MA; 7 Liver Care Network, Swedish Medical Center, Seattle, WA 

BACKGROUND: The current recommendation for the treatment 

of genotype 1 non-cirrhotic patients who are naïve to treatment 

is 12 weeks ledipasvir/sofosbuvir (LDV/SOF) though patients 

with baseline HCV RNA 2. SUMMARY: An examination of a 

real-world heterogeneous Hepatitis C population revealed a 

preference for 12 weeks even when baseline HCV RNA is 


combination therapy of Asunaprevir and Daclatasvir in chronic 

hepatitis C patients improved NK cell activity by increasing 

the frequency of CD56 bright NK cell in peripheral blood. It was 

revealed that reduction of HCV load by DAAs treatment in 

chronic hepatitis C patients could restore NK cell activity that 

was reduced by HCV. 

Disclosures: 

The following authors have nothing to disclose: Ikuo Nakamura, Yoshihiro Furuichi, 

Katsutoshi Sugimoto, Yoshiyuki Kobayashi, Fuminori Moriyasu 

1048 

Demographic Predictors of Direct Acting Antiviral 

Receipt in Individuals with Hepatitis C Virus Infection 

Fasiha Kanwal 1 , Jennifer R. Kramer 1 , Yumei Cao 1 , Thom Taylor 2 , 

Donna Smith 1 , Steven Asch 2 , Hashem B. El-Serag 1 ; 1 Michael E 

DeBakey VA and Baylor College of Medicine, Houston, TX; 2 VA 

Palo Alto Health Care System, Menlo Park, CA 

Background: Direct acting antiviral agents (DAAs) are highly 

effective yet very expensive. The extent to which the receipt of 

DAAs varies among socio-demographic groups is unknown. 

We sought to examine how race, gender, and age predicted 

receipt of DAAs in a U.S. cohort of HCV infected veterans. 

Methods: We used the Veterans Administration (VA) Corporate 

Data Warehouse to examine a cohort of patients with a 

positive HCV RNA test that had an outpatient visit to one of 

the VA facilities nationwide in 2014. We excluded patients 

if they had achieved sustained virological response prior to 

12/2013. DAA receipt included any prescription for sofosbuvir, 

simeprevir, Harvoni or Viekira from 12/1/2013 to 

2/10/2015. We examined demographic predictors of DAA 

receipt including age, race/ethnicity (Black, Hispanic, non-Hispanic 

white, others), and gender while adjusting for HCV genotype, 

HIV co-infection, diabetes, depression, alcohol or drug 

abuse, cirrhosis, and comorbidity using multivariable hierarchical 

logistic regression models. We also examined interactions 

between race/ethnicity and age or gender. Results: We had 

145,596 patients in our cohort. Their mean age was 60.1 

years (sd=7.3), 37.0% were Black, 47.8% non-Hispanic white, 

and 4.2% were Hispanic. Majority (96.7%) were males and 

62.0% had genotype 1, 7% genotype 2, 4.6% genotype 3, 

0.7% genotype 4; 25.7% did not have an HCV genotype test. 

In the cohort, 13,238 (9.1%) received DAAs. Black patients 

were significantly less likely to receive DAAs than non-Hispanic 

whites (OR=0.79; 95%CI: 0.74-0.84). There were no 

significant differences in DAAs receipt between non-Hispanic 

whites and patients of other races/ethnicities. Women were 

more likely to receive DAAs than men (OR=1.12; 1.02-1.24). 

Younger patients (


MELD Change from Baseline to Follow-up Week 12 












Gilead 

The following authors have nothing to disclose: Hans Van Vlierberghe, Robert 

Brown 

Disclosures: 








Novartis 









Jill M. Denning - Employment: Gilead Sciences, Inc. 

Sarah Arterburn - Employment: Gilead Sciences Inc.; Stock Shareholder: Gilead 

Sciences Inc. 



Sciences 


Sciences 

Princy N. Kumar - Consulting: Janssen, ViiV Healthcare; Grant/Research Support: 

Janssen, Merck, GSK, Gilead; Stock Shareholder: Merck, Phizer, Johnson&- 

Johnson, GSK, Gilead 


















David J. Mutimer - Advisory Committees or Review Panels: Gilead Sciences, 

AbbVie, Janssen, MSD, BMS; Speaking and Teaching: Gilead Sciences, AbbVie, 

Janssen, BMS 

Steven L. Flamm - Advisory Committees or Review Panels: Gilead, Bristol Myers 

Squibb, AbbVie, Janssen, Salix; Consulting: Merck, Janssen, Bristol Myers 

Squibb, AbbVie, Salix, Gilead; Grant/Research Support: Janssen, Bristol Myers 

Squibb, Gilead, AbbVie; Speaking and Teaching: Salix 






1050 

Adherence and Discontinuation Rates of Sofosbuvir-Based 

Regimens: Modeling Real World Experience in 

a Large Managed Care Organization 

Pravin S. Kamble 2 , David R. Walker 1 , Steven Marx 1 , Ray Harvey 2 , 

Claudia L. Uribe 2 , Suvapun Bunniran 2 , Jenna Collins 2 ; 1 HEOR, 

Abbvie, North Chicago, IL; 2 Outcomes, Comprehensive Health 

Insights, Louisville, KY 

BACKGROUND Discontinuation (DC) rates of sofosbuvir (SOF) 

based medications are generally very low in clinical trials (< 

1%). Adherence (ADH) rates on the other hand are rarely 

reported in clinical trials. Only a handful of studies have examined 

this issue outside of clinical trials, however, those studies 

did not comprehensively attempt to adjust for factors that may 

impact DC and ADH rates. OBJECTIVES The objective was to 

assess medication ADH and DC rates in SOF-based regimens 

and identify factors potentially associated with poor ADH and 

DC rates in patients with the hepatitis C virus (HCV). METH- 

ODS A retrospective cohort study using administrative claims 

data from a large managed care organization from May 2013 

to Sept 2014. Plan types included Medicare Advantage Prescription 

Drug (MAPD) and commercial (COM) which provide 

medical and pharmacy benefits. The study cohort included 

patients initiating 12 or 24-week treatment on SOF. The index 

date was defined as the first prescription fill date for SOF from 

Nov 2013 to March or May 2014 with a follow up period of 

12 or 24 weeks. Continuous enrollment for 6 months pre-index 

and 4 or 6 months post index date was required. ADH was calculated 

using proportion of days covered (PDC). Patients with 

at least 85% PDC were categorized as adherent. Patients were 

deemed discontinued if a gap of >14 days exists between 

fills. Regression analyses were conducted to identify baseline 

covariates associated with ADH or DC among plan members. 

Covariates included age, gender, risk score, plan type, geographic 

region, treatment co-pay, prior treatment experience, 

use of interferon, treatment duration and baseline healthcare 

use and costs. RESULTS In total, 514 MAPD and 63 COM 

members initiated HCV treatment with DAA. Patients were 63% 

male, mean age of 60 years, 84% on 12-week treatment, 

and 36% of SOF members were also on peginteferon. The 

discontinuation rate was 18% and the percent of members 

considered non-adherent was 14%. Based on the regression 

analysis, only older age, higher comorbidity risk score and 

use of peginterferon were significantly positively associated 

with non-adherence after controlling for other covariates. For 

12-week treatment patients, being in a non-HMO plan or in 

Medicare were predictors of being at lower risk of medication 

discontinuation. CONCLUSIONS In a real world setting, 14% 

of members on a new DAA are not adherent and 18% had 

fill gaps greater than 14 days. Sicker, older members are less 

likely to be adherent, whereas non-HMO and MAPD members


have lower risk of discontinuation. Additional patient support 

may be needed to optimize ADH in these patients. 

Disclosures: 

Pravin S. Kamble - Employment: Comprehensive Health Insights Inc. 

David R. Walker - Employment: Abbvie; Stock Shareholder: Abbvie, Baxter 

Healthcare 


Ray Harvey - Consulting: Comprehensive Health Insights 

Claudia L. Uribe - Employment: Humana 

Jenna Collins - Consulting: Comprehensive Health Insights, Humana 

The following authors have nothing to disclose: Suvapun Bunniran 

1051 

Turquoise-III: 12-Week Ribavirin-Free Regimen Of 

Ombitasvir/Paritaprevir/R And Dasabuvir For Patients 

With HCV Genotype 1B And Cirrhosis 

Fred Poordad 1 , Jordan J. Feld 2 , Roger Trinh 3 , Yves J. Horsmans 4 , 

Magdy Elkhashab 5 , Stefan Bourgeois 6 , Samuel S. Lee 7 , Christophe 

Moreno 8 , David Bernstein 9 , Ziad Younes 10 , Akshanth R. Polepally 3 , 

Kevin Howieson 3 , Bo Fu 3 , Greg Ball 3 , Nancy Shulman 3 , Edward 

Tam 11 ; 1 The Texas Liver Institute/University of Texas Health Science 

Center, San Antonio, TX; 2 Toronto Centre for Liver Disease, 

University of Toronto, Toronto, ON, Canada; 3 AbbVie, Inc., North 

Chicago, IL; 4 Université Catholique de Louvain, Brussels, Belgium; 

5 Toronto Liver Centre, Toronto, ON, Canada; 6 ZNA Stuivenberg, 

Antwerpen, Belgium; 7 University of Calgary, Calgary, AB, Canada; 

8 CUB Hôpital Erasme, Université Libre de Bruxelles, Brussels, 

Belgium; 9 North Shore University Hospital, Manhasset, NY; 10 GastroOne, 

Germantown, TN; 11 LAIR Centre, Vancouver, BC, Canada 

Introduction: Hepatitis C virus (HCV) infected patients have 

historically been more difficult to cure when they have cirrhosis. 

Treatment with the 3 direct-acting antiviral (3D) regimen 

of ombitasvir, paritaprevir (identified by AbbVie and Enanta, 

boosted with ritonavir), and dasabuvir without ribavirin (RBV) 

for 12 weeks has demonstrated 12-week sustained virologic 

response (SVR12) rates of 100% in HCV genotype (GT) 1b 

patients without cirrhosis, and 99% in GT1b patients with 

compensated cirrhosis when co-administered with RBV for 12 

weeks. We report the safety and efficacy of the 3D regimen 

without RBV in patients with HCV GT1b infection and compensated 

cirrhosis. Methods: Patients enrolled in this phase 

3b, multicenter, open-label study received 12 weeks of 3D 

without RBV. Both treatment-naïve and peginterferon/RBV treatment-experienced 

patients with compensated cirrhosis with no 

history of decompensation were enrolled with the following 

criteria: hemoglobin ≥10 g/dL, albumin ≥2.8 g/dL, platelet 

count ≥25 x 10 9 /L, and creatinine clearance ≥30 ml/min. 

Efficacy was assessed by the percentage of patients achieving 

SVR (HCV RNA below the level of quantitation [LLOQ; 


1052 

High prevalence of co-morbidities and complex polypharmacy 

with drug-drug interaction (DDI) potential in 

patients with chronic Hepatitis C (CHC). Consistent findings 

from large primary care databases in the United 

Kingdom, Germany, and France 

Fiona Marra 1,10 , Werner Leber 3 , Stephen T. Barclay 2 , Stefan 

Christensen 5 , Denis Ouzan 7 , Valerie Oules 8 , Peter S. McMahon 4 , 

Karel Kostev 6 , Xavier Ansolabehere 9 ; 1 NHS Greater Glasgow and 

Clyde, Glasgow, United Kingdom; 2 Walton Liver Clinic, Glasgow, 

United Kingdom; 3 Centre for Primary Care and Public Health, London, 

United Kingdom; 4 IMS Health UK, London, United Kingdom; 

5 Center for Interdisciplinary Medicine (CIM) Infectious Diseases, 

Münster, Germany; 6 IMS Health Germany, Frankfurt, Germany; 

7 Institut Arnault Tzanck, Saint-Laurent-du-Var, France; 8 Hôpital 

Saint Joseph, Marseille, France; 9 IMS Health France, Paris, France; 

10 University of Liverpool, Liverpool, United Kingdom 

Background Comorbidities and co-medications with DDI in 

patients with CHC may pose a challenge to upscaling treatment 

with new direct-acting antivirals (DAAs) and should be 

considered when selecting treatment options since DDI potential 

varies among regimens. We describe prevalence of co-morbidities 

and co-medications in large samples of CHC patients in 

primary care in the United Kingdom (UK), Germany (GER) and 

France (FRA). Methods Patients diagnosed with CHC between 

2011 and 04/2015 were identified using Read codes in the 

Clinical Practice Research Data link (UK) and ICD10 codes 

on the IMS® Disease Analyzer (GER/FRA). Inclusion criteria: 

18+ years, no evidence of sustained virological response, minimum 

one-year follow-up. Prescriptions within 12 months of 

last active date were categorized for DDI potential as either 

RED (contraindicated) or AMBER (additional monitoring/dose 

reduction required) with at least one of currently licensed DAA 

(OBV/PTV/r+DSV, SMV, LDV/SOF, DCV) using www.hep-druginteractions.org. 

Read/ICD10 coded co-morbidities were 

analysed. Comorbidities of interest included those that may 

make CHC treatment more difficult to manage. Results 7,949 

patients (UK: 4,644/GER: 2,737/FRA: 568) were analysed. 

Variation in mean age (45.8/51.8/57.1 years) suggests differences 

in CHC populations; 65%/58%/59% were male. 

Between 12%-19% of patients were on RED drugs, however 

only 0.9%-2.9% were on drugs contraindicated for all DAA 

regimen. 28.9%/35.1%/39.1% were on two or more RED 

or AMBER drugs, polypharmacy increased with age {18-29 

years (15.1%), 70+years (56.1%)}. Most common therapeutic 

classes for DDI are presented in table. Most common co-morbidities 

in the UK were: depression (23.3%), cardio-vascular 

disease [CVD] (21.0%) and COPD/Asthma (14.4%); compared 

to chronic pain syndromes (22.7%/16.6%) and hypertension 

(22.1%/15.1%) in GER/FRA, depression (14.7%) in 

GER and diabetes (8.7%) in FRA. Conclusion We observed 

significant co-morbidity and co-prescribing with DDI potential 

in CHC patients in three countries. The large difference in 

proportion of co-medications contraindicated to all DAA vs. 

only to some suggests careful selection of the DAA regimen is 

required. Treating patients at a younger age likely reduces the 

risk of DDI. 

RED or AMBER co-medication with at least one DAA in CHC 

patients in primary care (n=7,949) 

Disclosures: 

Fiona Marra - Advisory Committees or Review Panels: Gilead, AbbVie, Merck; 

Consulting: Gilead; Speaking and Teaching: Gilead, Abbvie, Merck, Janssen 

Stephen T. Barclay - Advisory Committees or Review Panels: Gilead, Janssen, 

MSD, Abbvie, BMS; Speaking and Teaching: Gilead, Janssen 

Stefan Christensen - Advisory Committees or Review Panels: BMS, Abbvie, Janssen, 

ViiV, Gilead, MSD; Speaking and Teaching: Gilead, MSD, Abbvie, BMS, 

ViiV, Reckitt Benckiser, Janssen 

Valerie Oules - Board Membership: Abbvie; Speaking and Teaching: gilead, 

BMS, Abbvie, Jansen 

Peter S. McMahon - Consulting: Gilead Sciences; Employment: IMS Health 

Karel Kostev - Consulting: IMS Health 

The following authors have nothing to disclose: Werner Leber, Denis Ouzan, 

Xavier Ansolabehere 

1053 

Real-Word Effectiveness of Ledipasvir (LDV)/Sofosbuvir 

(SOF)-Based Therapy for Hepatitis C Virus (HCV): 

Preliminary Analysis in a Large Integrated Health Care 

System 

Jennifer B. Lai 2,1 , David J. Witt 2 ; 1 Gastroenterology, Kaiser Permanente, 

San Rafael, CA; 2 Infectious Diseases, Kaiser Permanente 

Medical Center, San Rafael, CA 

BACKGROUND AND AIMS: Second generation direct-acting 

antiviral agents have become an integral component of treatment 

for HCV infection but little is known about their real-world 

effectiveness, particularly in community practice. We report 

virologic responses of patients with HCV genotype (GT) 1 infection 

receiving LDV/SOF with or without ribavirin (RBV) therapy. 

METHODS: Approval was obtained from the Kaiser Permanente 

Northern California Institutional Review Board. Treatment naïve 

and treatment experienced (TE) HCV GT 1 patients who started 

LDV/SOF-based therapy were identified by electronic prescription 

and medical records and analyzed on an intent-to-treat 

basis. End-of-Treatment (EOT) and Sustained Viral Response 4 

and 12 (SVR4, SVR12) were defined as HCV RNA less than 

the lower limit of quantification (LLOQ) upon completion of 

therapy or at 4 and 12 weeks after completing therapy, respectively. 

Liver fibrosis was assessed; patients were deemed to be 

(a) cirrhotic if either (i) any history of biopsy-proven cirrhosis, 

(ii) clinical manifestation(s) of cirrhosis or (iii) AST to Platelet 

Ratio Index (APRI) > 2.0 in the absence of biopsy ≤ 3 years 

prior to day 1 (D1) of treatment, (b) non-cirrhotic if either (i) 

no evidence of cirrhosis per biopsy ≤ 3 years prior to D1 or 

(ii) APRI < 0.5 in the absence of biopsy ≤ 3 years prior to 

D1 or otherwise (c) undetermined fibrosis. RESULTS: 656 GT1 

patients started LDV/SOF-based therapy at the time of data 

analysis of which 267 patients started LDV/SOF/RBV therapy 

for 12 weeks and 389 patients started LDV/SOF therapy for up 

to 24 weeks based on clinician discretion. Respective baseline 

characteristics were median age (61, 60 years), male (69, 

64%), TE (78, 16%) and cirrhotic (45, 23%), non-cirrhotic (19, 

37%) and undetermined fibrosis (36, 40%). We report preliminary 

EOT and SVR4 rates of 100% (173/173) and 98% 

(57/58) for patients receiving LDV/SOF/RBV therapy versus 

99% (223/226) and 94% (58/62) for patients receiving LDV/


SOF therapy, respectively. Additional virologic response data, 

including SVR12 responses, will be reported. CONCLUSIONS: 

We found high preliminary EOT and SVR4 rates for patients 

with HCV GT 1 treated with LDV/SOF-based therapy of 94 to 

100% which are very comparable to the 94 to 100% virologic 

responses reported in the published ION study series. 

Disclosures: 

The following authors have nothing to disclose: Jennifer B. Lai, David J. Witt 

1054 

Hepatitis C Cure Could Avoid Liver Transplant In Some 

Cirrhotic Patients On Dialysis Listed for Simultaneous 

Liver Kidney Transplantation 

Frank Czul, David Roth, Rodrigo M. Vianna, Cynthia Levy, Paul 

Martin, Kalyan R. Bhamidimarri; Hepatology, University of Miami- 

Miller School of Medicine, Miami, FL 

Background: Simultaneous liver kidney transplant (SLKT) is recommended 

in Hepatitis C (HCV) infected dialysis patients with 

cirrhosis but isolated kidney transplant (KT) can be considered 

in a subset that have early cirrhosis without portal hypertension. 

HCV cure could potentially avoid the need for liver transplantation 

in some patients listed for SLKT. Aim: To report our new 

strategy in SLKT wait-listed patients, whereby liver transplant 

(LT) was avoided in those who achieved HCV cure and did not 

demonstrate portal hypertension. Methods We describe our 

experience of 9 HCV infected patients with cirrhosis and end 

stage renal disease (ESRD) on hemodialysis who were initially 

listed for SLKT at the Miami Transplant Institute from 2011 to 

2014. Of the 9 patients, 6 (67%) were male with a mean age 

of 59.8 (+/-5.9) infected with HCV genotype 1 (78% GT1a). 

All of them underwent liver biopsy which confirmed the presence 

of cirrhosis, 8 (89%) Child A and 1 (11%) Child B, 1 

decompensated by small esophageal varices (EV) and 1 by EV 

+ ascites. The patients underwent HCV treatment, 2 (22%) with 

Pegylated interferon, Ribavirin and Boceprevir and 7 (78%) 

with Simeprevir and half dose Sofosbuvir. Results Among them, 

8 patients (89%) underwent trans-jugular liver biopsy with portal 

pressures measurements and 1 underwent percutaneous 

biopsy (see table). Seven (78%) patients achieved SVR12 and 

given the absence of portal hypertension, they were delisted 

from LT and were considered only for KT. The 2 patients who 

did not achieve SVR continued to be listed for SLKT. At the 

end of treatment there was a significant decrease on mean 

AST and ALT from 32.1 to 18.1 (p=0.047) and 31.6 to 15 

(p=0.0099). No other statistically significant differences were 

appreciated in the laboratory data. Conclusion HCV treatment 

could be offered to dialysis patients after they undergo KT. 

However, SLKT wait-listed patients could derive a huge benefit 

by pre-transplant HCV cure as LT could be avoided in those 

with compensated cirrhosis and devoid of portal hypertension. 

Characteristics of SLKT wait-listed patients 

Paul Martin - Advisory Committees or Review Panels: BMS; Grant/Research 

Support: Merck, Gilead, Janssen, Abbvie 

Kalyan R. Bhamidimarri - Advisory Committees or Review Panels: Gilead, Abb- 

Vie, Janssen; Grant/Research Support: Bristol Squibb Myers, Biotest, Synageva, 

Salix, Vital Therapies, Ocera Inc 

The following authors have nothing to disclose: Frank Czul, Rodrigo M. Vianna, 

Cynthia Levy 

1055 

Association of polymorphisms in the hepatitis C virus 

NS5B polymerase with reduced virologic response to 

sofosbuvir-based treatment 

Johannes Vermehren 1 , Simone Susser 1 , Christoph P. Berg 2 , Martin 

W. Welker 1 , Bernd Kronenberger 1 , Caterina Berkowski 1 , Stefan 

Zeuzem 1 , Christoph Sarrazin 1 ; 1 Medizinische Klinik 1, Klinikum 

der J. W. Goethe-Universität, Frankfurt am Main, Germany; 2 Universitätsklinikum 

Tübingen, Tübingen, Germany 

Background Sofosbuvir (SOF) is a nucleotide inhibitor of the 

hepatitis C virus (HCV) RNA polymerase NS5B that has been 

approved for the treatment of chronic HCV infection. SOF 

binds to the highly conserved active site of NS5B. Therefore, 

SOF has shown pangenotypic activity and a high barrier to 

resistance in vitro and in vivo. However, the importance of a 

number of minor variants within the HCV NS5B polymerase for 

treatment outcome of SOF-based therapy without other DAAs 

is unknown. Methods All consecutive patients with HCV GT1 

infection who received a SOF-based therapy with SOF being 

the only DAA were included in the study. Patients received 

either SOF in combination with PEG-IFN and ribavirin (RBV) 

or SOF in combination with RBV alone. Baseline samples were 

tested for the presence of resistance associated variants (RAVs) 

at positions L159, S282, C316, V321 und S368 of the NS5B 

polymerase by direct sequencing. The presence of these variants 

was tested for associations with treatment outcome. Results 

In total, 54 patients with HCV GT1 infection (24 with HCV 

GT1b) were treated with a SOF-based regimen (SOF+RBV: 

n=18; SOF+PEG-IFN+RBV: n=38). In patients with HCV GT1a 

and available follow-up data, 23/28 (82%) achieved a sustained 

virologic response (SVR) whereas only 14/23 (62%) 

patients with GT1b achieved an SVR. The C316N RAV was 

detected in 2 GT1a patients, both of whom did not achieve an 

SVR (1 relapse, 1 treatment discontinuation due to side effects). 

C316N was detected in 12 patients with GT1b, of whom 6 

achieved an SVR and 6 did not (all with relapse). In GT1b 

patients, SVR rates were significantly higher in patients without 

RAVs compared to patients in whom C316N was present 

(87.5% vs. 50%; p








The following authors have nothing to disclose: Simone Susser, Christoph P. Berg, 

Caterina Berkowski 

1056 

Hepatocellular carcinoma development in hepatitis C 

virus patients who achieved sustained viral response by 

interferon therapy: a large scale, long-term cohort study 

Yuko Nagaoki, Hiroshi Aikata, Hiromi Kan, Hatsue Fujino, Tomoki 

Kobayashi, Takayuki Fukuhara, Keiichi Masaki, Takashi Nakahara, 

Tomokazu Kawaoka, Masataka Tsuge, Akira Hiramatu, 

Michio Imamura, Yoshiiku Kawakami, Hidenori Ochi, Kazuaki 

Chayama; Department of Gastroenterology and Metabolism, Hiroshima 

University, Hiroshima, Japan 

Background We assessed risk factors for the development of 

hepatocellular carcinoma (HCC) following successful eradication 

of hepatitis C virus (HCV) with interferon therapy in a longterm, 

large-scale cohort study. Methods We reviewed 2262 

consecutive patients with HCV infection who achieved sustained 

viral response (SVR) by interferon therapy between January 

1995 and September 2013 in our study group. Among them, 

1094 patients who met the following inclusion criteria were 

enrolled in this retrospective cohort study: 1) received HCC 

surveillance by tumor markers (AFP, DCP), ultrasonography 

and/or dynamic computed tomography at least biannually; 2) 

underwent histological examination with liver biopsy before 

IFN therapy; and 3) with no evidence of HCC development 

prior to HCV eradication and within a year following eradication. 

We assessed host, pre-IFN treatment, and post-treatment 

risk factors associated with HCC development among these 

SVR patients. Results 1) Patients included 585 males aged 11 

to 85 years (median 60) at the time of HCV eradication. 581 

patients had HCV genotype 1b, and 513 had genotypes 2a, 

2b or other genotypes. 936 patients had interferon lambda 3 

(IFNL3) SNP rs8099917 genotype TT, and 158 had genotypes 

GG or TG. 885 patients had DEPDC5 SNP rs1012068 genotype 

TT, and 209 had genotype GG or TG. Histological fibrosis 

stage before IFN treatment was as follows: F0 or F1, 492; 

F2, 39; F3, 174; and F4, 34. 2) During the observation period 

(median 50 months: range 13-224), 36 (3%) of 1094 patients 

developed HCC after HCV eradication. The median period 

from HCV eradication to HCC development was 37 months 

(range 17-141), and the cumulative rates of developing HCC 

at 5,10 and 15 years were 4%, 6% and 12%, respectively. 3) 

As independent risk factors for HCC development, multivariate 

analysis identified older age ≥ 60 years (HR, 3.1: 95%CI, 

1.3-6.6: P=0.009), male sex (HR, 11.9: 95%CI, 2.8-50.0: 

P


Treatment Outcomes 

N* number of patients with available SVR12 

Disclosures: 











Alexander Kuo - Advisory Committees or Review Panels: Gilead; Grant/Research 

Support: Gilead; Speaking and Teaching: IAS-USA 




Jama M. Darling - Consulting: BristolMyers Squibb; Grant/Research Support: 

BristolMyers Squibb 





Lynn M. Frazier - Advisory Committees or Review Panels: Abbvie ; Speaking and 

Teaching: Jansen, Gilead 

Saleh Alqahtani - Advisory Committees or Review Panels: Gilead Sciences, Janssen 

Therapeutics; Consulting: Abbvie; Grant/Research Support: Merck & Co, 

Inc., Abbvie, Gilead 



BMS 






The following authors have nothing to disclose: Giuseppe Morelli, Joseph S. 

Galati 

1058 

Daclatasvir plus sofosbuvir with or without ribavirin for 

the treatment of chronic HCV in patients coinfected with 

HIV: Interim results of a multicenter compassionate use 

program 

Jürgen K. Rockstroh 1 , Tania M. Welzel 2 , Patrick Ingiliz 3 , Joerg 

Petersen 4 , Marc van der Valk 5 , Kerstin Herzer 6 , Peter Ferenci 7 , 

Michael Gschwantler 8 , Markus Cornberg 9 , Thomas Berg 10 , Ulrich 

Spengler 1 , Ola Weiland 11 , Hartwig Klinker 12 , Markus Peck-Radosavljevic 

7 , Yue Zhao 13 , Maria Jesus Jimenez Exposito 14 , Stefan 

Zeuzem 2 ; 1 Universitätsklinikum Bonn, Bonn, Germany; 2 Universitätsklinikum 

der Johann Wolfgang Goethe Universität, Frankfurt, 

Germany; 3 Praxis Driesener Strasse, Berlin, Germany; 4 IFI Institut 

für Interdisziplinäre Medizin, Hamburg, Germany; 5 Academic 

Medical Center, University of Amsterdam, Amsterdam, Netherlands; 

6 Universitätsklinikum Essen (AöR), Essen, Germany; 7 Medizinische 

Universität Wien, Vienna, Austria; 8 Wilhelminenspital, 

Vienna, Austria; 9 Medizinische Hochschule Hannover, Hannover, 

Germany; 10 Universitätsklinikum Leipzig, Leipzig, Germany; 

11 Karolinska Institutet, Karolinska University Hospital Huddinge, 

Stockholm, Sweden; 12 Universitätsklinikum Würzburg, Würzburg, 

Germany; 13 Global Biostatistics, Bristol-Myers Squibb, Hopewell, 

NJ; 14 Research and Development, Bristol-Myers Squibb, Princeton, 

NJ 

BACKGROUND: HCV-related liver disease is a leading cause 

of death among HIV/HCV coinfected patients. Interferon-free 

regimens that can be safely co-administered with antiretroviral 

(ARV) therapy are needed in this population. In clinical trials 

(ALLY-2 study), the pangenotypic, all-oral 12-weeks regimen of 

daclatasvir (DCV) and sofosbuvir (SOF) was well tolerated and 

achieved 97% SVR rates in HIV/HCV coinfected patients receiving 

a wide range of ARVs. We report here interim results on the 

combination DCV plus SOF, with or without ribavirin (RBV) in 

HIV/HCV coinfected patients enrolled in a compassionate use 

program (CUP). METHODS: This CUP enrolled adult patients 

with chronic HCV infection who were at a high risk of hepatic 

decompensation or death within 12 months if left untreated, 

and who had no available treatment options. Patients received 

DCV 60 mg + SOF 400 mg QD, with RBV added at the physician’s 

discretion. Dose adjustments for DCV were allowed for 

concomitant use of ARVs: 30 mg with ritonavir-boosted PIs; 90 

mg with NNRTIs, except rilpivirine. Recommended duration 

of treatment was 24 weeks. This interim analysis includes 55 

HCV-HIV coinfected patients enrolled in the CUP with available 

data on March 16, 2015, of whom 15 have available SVR12 

data to date. RESULTS: Most patients were male (82%), white 

(95%), with a median age of 52 years (31–67). HIV RNA 

was undetectable in 18/21 patients; median baseline HIV 

RNA: 20 copies/mL (0-100 copies/mL); 17/27 patients had 

a CD4 cell count was


patients in a real-world setting, DCV+SOF±RBV demonstrated 

good rates of SVR12 and was well tolerated in a HCV-HIV 

coinfected patient population containing a high proportion of 

cirrhotic patients. 

Disclosures: 






































The following authors have nothing to disclose: Patrick Ingiliz, Kerstin Herzer, 

Ulrich Spengler, Hartwig Klinker, Yue Zhao 

1059 

The Significance of Immunoallegic Adverse Events 

Developing during Dual Oral Therapy with Daclatasvir 

Plus Asunaprevir in Patients with Genotype 1b HCV 

Infection 

YOHEI FUJII, Yoshihito Uchida, Kayoko Sugawara, Mie Inao, 

Nobuaki Nakayama, Satoshi Mochida; SAITAMA MEDICAL UNI- 

VERSITY, Saitama, Japan 

Aim: Dual oral therapy with daclatasvir (DCV) plus asunaprevir 

(ASV) was approved in Japan in July 2014 for patients 

with genotype 1b HCV. We established a novel system to 

quantify NS5A-RAVs using cycling-probe real-time PCR (PLos 

One, 2014), and performed the therapy by priority for patients 

without such RAVs. Although SVR4-12 was achieved in 95% of 

DAA-naïve patients without baseline NS5A-RAVs, severe immunoallegic 

hepatitis resembling drug-induced hypersensitive syndrome 

(DIHS) developed in one patient (Hepatology 2015; 

61: 400). Thus, the significance of immunoalergic adverse 

events during DCV/ASV therapies were evaluated. Methods 

& Results: A total of 224 patients (98 men. 126 women; 23 

to 87 years old.) received DCV/ASV administrations for 24 

weeks since Semtember 2014. Adverse events were seen 

during the therapy in 97 patients (43%); liver injury in 33 

(15%), eosinophilia in 31 (14%), fever higher than 38C in 22 

(10%) and decompasation of cirrhosis in 3 (1%). Among them, 

the therapy was discontinued in 18 patients (8%) including 8 

patients showing grade-4 elevation of serum ALT levels (>300 

IU/L) and 1 patient with increase of both serum bilirubin concentrations 

and GGT and ALP levels. Fever occurred between 

7 and 14 days after the initiation of therapy simultaneously 

with serum CRP level elevation, but was attenuated after oral 

intakes of NSAIDs for between 3 and 7 days, and did not 

recur following discontinuation of NSAIDs intakes. Among 33 

patients showing liver injuries, serum CRP level elevation and/ 

or eosinophilia were seen simultaneously in 4 patients; 3 and 

1 patient(s) showing elevation of serum ALT levels and bilirubin 

concentrations, respectively, and oral and/or intravenous glucocorticoid 

adminstrations were required in 3 patients, since 

the events were not attenuated following discontinuation of 

dual oral therapy. In remaining 29 patients showing serum ALT 

level elevation, the therapy was continued for 24 weeks in 21 

patients, while dairy doses of ASV were reduced from 200 mg 

to 100 mg in 8 patients to maintain serum ALT levels less than 

150 IU/mL. Conclusion: Immunoallegic adverse events such as 

fever, serum CRP level elevation and eosinophilia occurred in 

frequent during dual oral therapy with DCV plus ASV, but were 

tolerable by NSAIDs intakes. Also, liver injuries developed frequently 

in these patients probable due to toxic effects by ASV. 

It should be noted, however, there existed patients showing 

immunoallergic liver injuries, presenting elevation of serum ALT 

levels and/or bilirubin concentrations combined with increased 

serum CRP levels and eosinophilia, since glucocorticoid administrations 

may be required for such patients. 

Disclosures: 

Yoshihito Uchida - Patent Held/Filed: SRL Inc. 

Satoshi Mochida - Grant/Research Support: Chugai, MSD, Tioray Medical, 

BMS; Speaking and Teaching: MSD, Toray Medical, BMS, Tanabe Mitsubishi 

The following authors have nothing to disclose: YOHEI FUJII, Kayoko Sugawara, 

Mie Inao, Nobuaki Nakayama 

1060 

Sofosbuvir-based treatments for patients with hepatitis 

C virus (HCV) mono-infection and human immunodeficiency 

virus (HIV)-HCV co-infection with genotype 3 in 

clinical practice – Results from the GErman hepatitis C 

COhort (GECCO) 

Patrick Ingiliz 1 , Knud Schewe 2 , Thomas Lutz 3 , Christoph Boesecke 4 , 

Stefan Mauss 5 , Heiner W. Busch 6 , Axel Baumgarten 1 , Marcel 

Schuetze 1 , Guenther Schmutz 5 , Stefan Christensen 6 , Karl-Georg 

Simon 7 , Dietrich Hueppe 8 ; 1 Medical Center for Infectious Diseases, 

Berlin, Germany; 2 ICH, Hamburg, Germany; 3 Infektiologikum, 

Frankfurt, Germany; 4 Department of Medicine I, University 

of Bonn, Bonn, Germany; 5 Center for HIV and Hepatogastroenterology, 

Duesseldorf, Germany; 6 Center for Interdisciplinary 

Medicine, Muenster, Germany; 7 Practice for Gastroenterology 

Leverkusen,, Leverkusen, Germany; 8 Center for Gastroenterology, 

Herne, Germany 

Introduction The first direct acting antivirals (DAA) were 

approved in Europe in 2014 based on limited study data. 

In particular, HIV-HCV co-infected patients and pre-treated 

patients had not been systematically studied. Here, we present 

real-life data on efficacy and safety of sofosbuvir (SOF)-based 

therapies from Germany in genotype (GT) 3 patients. Methods 

In this multicenter cohort, all patients who were started 

on sofosbuvir-based treatments were documented. For the current 

analysis, only patients with HCV genotype (GT) 3 on the 

following regimes were included: SOF/PegIFN/RBV (ribavirin)/12 

weeks, SOF/RBV/12 or 24 weeks, SOF/DCV (daclatasvir)/12 

or 24 weeks, and SOF/LDV (ledipasvir)/12 or 24 

weeks. All patients within the GECCO cohort are part of the


prospective German hepatitis C registry. Results Overall, 836 

patients on SOF-containing regimens have been enrolled so 

far, 629 (75%) are HCV-monoinfected and 207 (25%) HIV- 

HCV co-infected. Of those, 155 (19%) patients were infected 

with HCV genotype 3, 120/155 (77%) were male. The median 

age (IQR) was 50 years (41 to 55). 65 patients (41%) have 

been unsuccesfully treated before, mainly with dual therapy 

consisting of pegylated interferon and ribavirin. Liver cirrhosis 

was diagnosed in 55 patients (37%), two patients were Child- 

Pugh stage B. The sustained virological response (SVR12) rate 

in patients treated with SOF/PegIFN/RBV (n=40) was 94% 

(2/31). In patients treated with SOF/RBV (n=76), the SVR12 

rate was 91% (21/23). In patients treated with SOF/DCV 

(n=17) for 12 or 24 weeks or SOF/LDV for 12 or 24 weeks, 

only very few patients have reached the SVR12 timepoint but all 

responded (4/4). Prior treatment failure did not affect treatment 

responses. Patients with liver cirrhosis had a lower response 

rate of 86% vs 97% without cirrhosis (p=0.28). Conclusion: In 

this large cohort patients with genotype 3 response rates were 

promising. Patients with liver cirrhosis responded less well to 

the different regimens used. This real-life data underline that 

the existing drug combinations are effective tools against HCV 

genotype 3. 

Disclosures: 

Patrick Ingiliz - Consulting: Gilead, Abbvie, Janssen Cilag; Speaking and Teaching: 

BMS, MSD, Gilead, Abbvie 

Knud Schewe - Advisory Committees or Review Panels: abbvie, gilead, msd, 

bms, janssen cilag, viiv, hexal 

Thomas Lutz - Advisory Committees or Review Panels: Gilead, MSD, AbbVie, 

BMS, Janssen Cilag; Grant/Research Support: Gilead, GlaxoSmithKline, Roche, 

MSD, AbbVie, Janssen Cilag; Speaking and Teaching: ViiV, BMS 

Christoph Boesecke - Speaking and Teaching: MSD, Boehringer, Gilead, BMS, 

Roche, Abbvie, ViiV 

Stefan Mauss - Advisory Committees or Review Panels: BMS, AbbVie, Janssen, 

ViiV, Gilead; Speaking and Teaching: BMS, AbbVie, Janssen, Gilead, MSD 

Heiner W. Busch - Speaking and Teaching: Tibotec, MSD, Janssen, BMS, Gilead 




Dietrich Hueppe - Advisory Committees or Review Panels: Roche, MSD, Novatis, 

Gilead, BMS, Janssen 

The following authors have nothing to disclose: Axel Baumgarten, Marcel Schuetze, 

Guenther Schmutz, Karl-Georg Simon 

1061 

A comparison of direct sequencing and PCR- invader 

assay for the detection of NS5A Y93H mutation and 

response to daclatasvir and asunaprevir therapy in 

patients with hepatitis C virus genotype 1b 

Kazuhiko Hayashi, Masatoshi Ishigami, Yoji Ishizu, Teiji Kuzuya, 

Takashi Honda, Yoshiki Hirooka, Hidemi Goto; Gastroenterology, 

Nagoya University, Nagoya, Japan 

OBJECTIVES: Interferon free regimen such as daclatasvir (DCV) 

and asunaprevir (ASV) was approved for patients with hepatitis 

C virus (HCV) genotype 1b. Virologic failure of DCV and 

ASV therapy was associated with baseline NS5A polymorphism 

such as Y93H mutation and screening for the presence 

of Y93H mutation at pretreatment is recommended. The direct 

sequencing analysis is standard method to identify Y93H mutation 

but needs a lot of efforts and costs. A simple assay based 

on the Q-Invader technology was developed to determine and 

quantify NS5A Y93H mutant strains with high sensitivity. This 

study sought to compare two methods used to detection of 

Y93H mutation and evaluate the effect of Y93H mutation on 

response to DCV and ASV therapy. METHODS: 121 patients 

with HCV genotype 1b were enrolled. All patients were examined 

the NS5A Y93H mutation by both direct sequencing 

analysis and PCR- invader assay. 45 patients were received 

DCV and ASV combination therapy for 24 weeks and were 

selected for virologic response study. RESULTS: 112 of 121 

(92.7%) patients were able to measure Y93H mutation by 

direct sequencing analysis and 109 of 121 (90.1%) patients 

were able to measure Y93H mutation by PCR- invader assay. 

There are no statistically significant differences of sensitivity 

in both methods. 21 of 112 (18.6%) patients were identified 

Y93H mutation strain by direct sequencing analysis and 18 of 

109 (16.5%) patients were able to detect Y93H mutation by 

PCR- invader assay. Twelve patients were defined with Y93H 

mutation by both methods and the percent HCV-RNA level of 

the Y93H mutant were 10, 24, 53, 55, 85, 89, 89, 95, 98, 

99, 99, and 99, respectively. Six patients were detected Y93H 

mutation by PCR- invader assay but failed to be detected by 

direct sequencing and the percent of the Y93H mutant were 4, 

5, 14, 30, 52, and 83, respectively. The difference in detection 

rate of Y93H mutation between two methods indicated that 

direct sequencing analysis could not measure less than 20% 

of minor strains. Nine cases were detected Y93H mutation by 

direct sequencing but failed to be detected by PCR- invader 

assay. One was Y93C, 3 were Y93H, and 5 were mixed type. 

The patients without Y93H mutation by both methods were 

achieved to SVR except for one case (97.1%). The other virologic 

failure were occurred in patients with Y93H mutation by 

either method. CONCLUSIONS: The sensitivity of PCR- invader 

assay was as same as that of direct sequencing analysis, but 

PCR- invader assay could measure minor strains not possible by 

direct sequencing analysis. A combination of direct sequencing 

and PCR- invader assay would improve prediction of the 

response to DCV and ASV therapy. 

Disclosures: 



The following authors have nothing to disclose: Kazuhiko Hayashi, Masatoshi 

Ishigami, Yoji Ishizu, Teiji Kuzuya, Takashi Honda, Yoshiki Hirooka 

1062 

Prediction for poor virological response and clinical 

utility of resistance-associated variant detection in combination 

therapy with direct-acting antivirals for HCV 

genotype 1 

Norio Akuta, Fumitaka Suzuki, Yusuke Kawamura, Hitomi Sezaki, 

Yoshiyuki Suzuki, Tetsuya Hosaka, Masahiro Kobayashi, Satoshi 

Saitoh, Mariko Kobayashi, Yasuji Arase, Kenji Ikeda, Hiromitsu 

Kumada; Hepatology, Toranomon Hospital, Tokyo, Japan 

BACKGROUND: All-oral combinations of direct-acting antivirals 

improved treatment efficacy for patients with HCV infection, 

but prediction of poor virological response and clinical utility 

of resistance-associated variant (RAV) detection are not clear. 

METHODS: 405 interferon-ineligible/intolerant and 340 nonresponder 

patients with chronic HCV genotype 1b infection 

introduced daclatasvir 60 mg once daily plus asunaprevir 100 

mg twice daily for 24 weeks. 44 patients, who failed to triple 

therapy of peginterferon plus ribavirin with NS3/4A protease 

inhibitor (27 patients of telaprevir, 15 of simeprevir, and 2 of 

simeprevir after telaprevir), were included. Sustained virologic 

response 24 weeks after treatment (SVR24) and non-virological 

response (patients who do not achieve HCV-RNA negativity 

during treatment) were evaluated. Patients with virological failure 

were tested for RAVs (NS3 substitutions of Q80, D168, 

and NS5A substitutions of L31, Y93) by direct sequencing at 

the baseline. NS5A-Y93 mutant at baseline was also measured 

by comparative quantitative analysis of the Q-Invader assay. 

RESULTS: SVR24 was evaluated in 62 patients, and the rates


were 71% of interferon-ineligible/intolerant patients and 89% 

of prior non-response patients. Non-virological response was 

evaluated in 583 patients, and the rates were 3%. The higher 

levels of NS5A-Y93 mutant affected the higher rates of non-virological 

response, significantly. Especially, in patients of IL28B 

rs8099917 non TT with NS5A-Y93 mutant levels 16 on 

CES-D) recorded in 44%. Average medication adherence was 

97%. Among patients who used drugs in the past month and 

year, adherence was 92% and 96%, respectively. Medication 

adherence in those who drank alcohol in past year was 97%. 

Nonadherence was due to missing doses (67%) and potential 

double-dosing (33%). Clinic attendance to scheduled visits was 

95% (range: 87% - 100%). All 32 patients (100%) completed 

the full duration of 12 weeks of therapy. At 12-week EOT visit, 

97% had HCV RNA below the level of quantitation. Conclusion: 

Rates of medication adherence, clinic attendance, and 

persistence are extremely high in patients with SUDs/AUDs. 

These data provide no justification for withholding treatment 

from these patients due to adherence concerns. Clinicians 

and patients need to be aware of accidental double-dosing. 

Research is needed to determine the extent to which financial 

incentives are needed to optimize adherence and to identify a 

new adherence threshold that may jeopardize viral cure. 

Disclosures: 

The following authors have nothing to disclose: Donna M. Evon, Angela Edwards, 

Becky Straub, Christopher B. Hurt, Harsha Thirumurthy, David Wohl 

1064 

Frequent Emergences of Rare RAVs Showing Extreme 

Resistance to NS5A Inhibitors during Dual Oral Threpy 

with Daclatasvir Plus Asunaprevir in Patients Previously 

Receiving Triple Therapy with Simeprevir 

Yoshihito Uchida, Nobuaki Nakayama, Jun-ichi Kouyama, Kayoko 

Naiki, Hayato Uemura, Kayoko Sugawara, Mie Inao, Satoshi 

Mochida; Department of Gastroenterology and Hepatology, 

Saitama Medical University, Iruma-gun, Saitama, Japan 

Aim: Dual oral therapy with daclatasvir (DCV) plus asunaprevir 

(ASV) was approved in Japan for patients with genotype 1b 

HCV in July 2014. We established a novel system to quantify 

NS5A-RAVs using cycling-probe real-time PCR (PLos One 

2014), and performed the therapy by priority for these without 

baseline RAVs. However, rare RAVs showing extreme resistance 

to NS5A inhibitors developed frequently during DCV/ 

ASV administrations especially in those with preveious triple 

therapy with simeprevir (SMV) despite that baseline RAVs were 

absent. We report on this serious problem. Methods: A total 

of 224 patients (98 men. 126 women; 23 to 87 years old.) 

received DCV/ASV for 24 weeks. Among them, 5 patients had 

a history of virologic failure after combined SMV/Peg-IFN/ 

ribavirin therapy. RAVs in NS3 and NS5A regions at baseline 

and during the therapies were evaluated by cycling-probe realtime 

PCR combined with direct sequencing. Results: Baseline 

Y93H and L31M mutations were found in 27 (12%) and 6 

(2%) patients, respectively, while those with both mutations 

were absent. The therapy has already been completed in 156 

patients, and SVR4-12 was achieved in 134 patients (86%); 

117 (91%) of 129 without NS5A-RAVs, 11 (52%) of 21 with 

Y93H mutation and all of 6 with L31M mutations. Baseline 

NS5A-RAVs were absent in 5 patients with previous therapies 

including SMV, but virologic failure developed in all of them; 

DCV/ASV was discontinued at 4 weeks in 2 due to null-response, 

at 8 weeks in 2 due to virologic rebound and at 20 

weeks in 1 due to breakthrough. Among them, both Y93H and 

L31M mutations were detected only in 1 patient showing virologic 

rebound, while were absent in the remaining 4 patients.


In these patients, rare mutations in NS5A region, R30H, A92K, 

P29del and P32del, were detected simultaneously with NS3- 

D168E/V mutations in HCV straines obtained at virologic failure. 

Especially in 2 patients showing null-response, P29del 

and P32del mutations were found, and experiments using replicon 

system revealed HCV strains with such deletions showed 

>390,000-fold registance against DCV. Consequently, SVR4- 

12 rates in patients without baseline Y93H mutation were 95% 

when those receiving SMV were excluded. Deep-sequencing 

to evaluate the kinetics of HCV strains with P29del and P32del 

was currently in progress. Conclusion: Although high SVR rates 

were obtained after DCV/ASV therapy in patients without 

baseline NS5A-RAVs, rare RAVs extremely resistant to NS5A 

inhibitors emerged frequently in those receiving SMV previously. 

Such patients should be add to the lists of contraindication 

for DCV/ASV therapy even when baseline NS5A-RAVs 

were absent. 

Disclosures: 


Jun-ichi Kouyama - Patent Held/Filed: SRL Inc. 

Kayoko Naiki - Patent Held/Filed: SRL Inc. 



The following authors have nothing to disclose: Nobuaki Nakayama, Hayato 

Uemura, Kayoko Sugawara, Mie Inao 

pts without cirrhosis received 12 weeks (wks) of 3D+RBV and 

cirrhotics received 12 or 24 wks of 3D+RBV. GT1b-infected 

pts without cirrhosis received 12 wks of 3D and cirrhotics 

received 12 wks of 3D+RBV. End of treatment response (EOTR) 

is reported; SVR12 will be presented. Results: 615 pts enrolled 

and received study drug. Baseline characteristics and treatment 

response are presented below. 98.0% (603/615) of pts 

achieved EOTR. 0.8% (5/615) of pts experienced on-treatment 

virologic failure, and 1.0% (6/615) discontinued treatment 

due to adverse events. The most common adverse events 

(occurring in >10% of pts) were fatigue, nausea, headache, 

insomnia, and pruritus. Serious adverse events considered at 

least possibly related to 3D treatment were reported in 0.5% 

(3/615) of pts. Conclusions: In the TOPAZ-II study, 3D+/-RBV 

achieved high rates of on-treatment response and has been 

well-tolerated with a low rate of discontinuation due to adverse 

events in a broad population of HCV GT1-infected pts. 

Baseline characteristics and efficacy 

1065 

Preliminary Safety and Efficacy Results from TOPAZ-II: 

A Phase 3b Study Evaluating Long-Term Clinical Outcomes 

in HCV Genotype 1-infected Patients Receiving 

Ombitasvir/Paritaprevir/r and Dasabuvir +/-Ribavirin 

Nancy Reau 1 , Fred Poordad 2 , Jeffrey Enejosa 3 , Asma Siddique 4 , 

Humberto I. Aguilar 5 , Jacob P. Lalezari 6 , Franco Felizarta 7 , Peter 

J. Ruane 8 , Peter Varunok 9 , David Bernstein 10 , Douglas Dieterich 11 , 

Gregory T. Everson 12 , Yiran Hu 3 , Greg Ball 3 , Tami Pilot-Matias 3 , 

Nancy Shulman 3 , Sanjeev Arora 13 ; 1 University of Chicago Medical 

Center, Chicago, IL; 2 The Texas Liver Institute/University of 

Texas Health Science Center, San Antonio, TX; 3 AbbVie Inc., 

North Chicago, IL; 4 Virginia Mason Medical Center, Seattle, WA; 

5 Louisiana Research Center, LLC, Shreveport, LA; 6 Quest Clinical 

Research, San Francisco, CA; 7 Private practice, Bakersfield, CA; 

8 Ruane Medical and Liver Health Institute, Los Angeles, CA; 9 Premier 

Medical Group of the Hudson Valley, PC, Poughkeepsie, NY; 

10 North Shore University Hospital, Manhasset, NY; 11 Mount Sinai 

School of Medicine, Icahn School of Medicine, New York, NY; 

12 University of Colorado Denver School of Medicine, Aurora, CO; 

13 University of New Mexico, Albuquerque, NM 

Background: The 3-direct-acting antiviral regimen of ombitasvir/paritaprevir/ritonavir 

(paritaprevir identified by AbbVie 

and Enanta, co-dosed with ritonavir) and dasabuvir (3D) +/-ribavirin 

(RBV) has resulted in high SVR12 rates with a favorable 

safety profile in HCV genotype (GT) 1-infected patients (pts), 

including those with compensated cirrhosis, in phase 3 trials. 

TOPAZ-I (ex-US) and TOPAZ-II (US) are evaluating the impact 

of SVR12 on progression of liver disease through 5 years 

post-treatment in a broad population of HCV GT1-infected pts 

receiving 3D+/-RBV. Here, we report on-treatment safety and 

efficacy of 3D+/-RBV among pts in the TOPAZ-II study. Methods: 

TOPAZ–II is a phase 3b, open-label, non-randomized, 

multicenter trial conducted in 48 community and academic 

centers in the US. Pts are treatment-naïve or interferon/RBV 

treatment-experienced, with or without compensated cirrhosis. 

There was no upper limit on age, BMI, or ALT/AST levels. Pts 

with creatinine clearance ≥30 mL/min, albumin ≥2.8 g/dL, 

and platelet count ≥25 × 10 9 /L were eligible. GT1a-infected 

*Data missing for 1 pt 

Disclosures: 









Jeffrey Enejosa - Employment: AbbVie; Stock Shareholder: AbbVie 

Humberto I. Aguilar - Speaking and Teaching: abbvie, gilead 

Franco Felizarta - Grant/Research Support: AbbVie, Gilead, Janssen, Merck, 

BMS, Boehringer-Ingelheim, Vertex, Roche; Speaking and Teaching: AbbVie, 

Gilead, Janssen, Merck 

Peter J. Ruane - Advisory Committees or Review Panels: Gilead, Boehringer, 

Jannsen, Viiv, Abbott; Consulting: Gilead, Jannsen, Abbott, BMS; Grant/ 

Research Support: Gilead, Boehringer, Jannsen, Idenix, Abbott, BMS; Speaking 

and Teaching: Gilead, Merck, Boehringer, Jannsen, VIIV, Abbott, BMS; Stock 





Gilead 











Yiran Hu - Employment: AbbVie Inc. 



The following authors have nothing to disclose: Asma Siddique, Jacob P. Lalezari, 

Peter Varunok, Greg Ball, Sanjeev Arora


1066 

Effect of Renal Function on the Pharmacokinetics of 

Ombitasvir/ Paritaprevir/Ritonavir, Dasabuvir and Ribavirin 

in Over 2000 Subjects with HCV GT1 Infection 

Akshanth R. Polepally 1 , Prajakta Badri 1 , Doerthe Eckert 2 , Sven 

Mensing 2 , Rajeev Menon 1 ; 1 Clinical Pharmacology and Pharmacometrics, 

AbbVie Inc.,, North Chicago, IL; 2 Clinical Pharmacology 

and Pharmacometrics, AbbVie Deutschland GmbH & Co.KG, 

Ludwigshafen, Germany 

Background and aims: Paritaprevir, a NS3/4A protease inhibitor 

(administered with low-dose ritonavir [RTV], paritaprevir/r), 

identified by AbbVie and Enanta, ombitasvir, a NS5A inhibitor, 

and dasabuvir, a NS5B polymerase inhibitor, are directly 

acting antivirals (DAAs) approved in combination (3D regimen) 

± ribavirin (RBV) for the treatment of HCV GT1 infection . The 

objective of this analysis was to evaluate the effect of renal function 

as estimated by creatinine clearance (CrCL) on the pharmacokinetics 

of the DAAs, RTV and RBV in HCV infected GT1 

subjects. Methods: Total exposure measured by area under the 

plasma concentration curve (AUC) was generated for DAAs, 

RTV and RBV using population pharmacokinetic modeling by 

pooling data from 6 phase 3 studies and 1 phase 2 study in > 

2000 HCV GT1 infected subjects. All subjects received ombitasvir/ 

paritaprevir/r 25/150/100 mg QD and dasabuvir 

250 mg BID ± weight based RBV. DAA and RTV AUC values 

were available from 2093 subjects and RBV AUC values were 

available from 1584 subjects. The dataset included subjects 

with normal renal function (NF) (CrCl ≥ 90 mL/min, n = 1495), 

mild renal impairment (RI) (CrCL 60-89 mL/min, n = 576) and 

moderate RI (CrCL 30-59 mL/min, n = 22). The effect of CrCL 

on the AUC values of each DAA, RTV and RBV was evaluated, 

and adjusted for any significant subject-specific covariates (at 

a significance level of 0.05) including, age, sex, body weight 

(BW), cirrhosis (CRHS) and Asian ethnicity (ASN) in multiple 

linear regression (MLR) analysis (R 3.2.0). CrCL was retained 

in the models, regardless of its statistical significance, to determine 

the effect, if any, on the AUC values. Using the final 

MLR model, AUC values were predicted for subjects with NF 

(CrCL=105 mL/min), mild RI (CrCL=75 mL/min) and moderate 

RI (CrCL=45 mL/min). Results: CrCL was not a statistically 

significant predictor of DAAs and RTV AUC values (p > 0.05). 

Age, sex, CRHS were significant covariates for all DAAs/RTV 

while BW and ASN were for ombitasvir and dasabuvir. CrCL 

showed a significant relation with the RBVAUC values (p < 

0.05), which is consistent with RBV’s predominant renal excretion. 

Age, sex, BW and CRHS were significant covariates for 

RBV. The DAA AUC values were comparable (≤ 10% difference) 

amongst different levels of renal function, while RBV AUC 

values were up to 17% higher in mild/moderate RI compared 

to NF. Conclusions: In HCV GT1-infected subjects with or without 

cirrhosis, mild/moderate RI did not affect DAA and RTV 

exposures; thus, no dose-adjustments are needed for the 3D 

regimen. RBV doses should be adjusted for renal impairment 

as recommended in its label. 

Disclosures: 

Akshanth R. Polepally - Employment: AbbVie 

Prajakta Badri - Employment: Abbvie; Stock Shareholder: Abbvie 

Sven Mensing - Employment: AbbVie 

Rajeev Menon - Employment: AbbVie; Stock Shareholder: AbbVie 

The following authors have nothing to disclose: Doerthe Eckert 

1067 

Clinical Management of Ribavirin Dosing in HCV-Infected 

Patients with Anemia-Related Events Receiving 

Ombitasvir/Paritaprevir/r and Dasabuvir 

Jordan J. Feld 2 , David Bernstein 3 , Ziad Younes 4 , Hans Van Vlierberghe 

5 , Greg Ball 1 , Ronald D’Amico 1 , Peter Ferenci 6 ; 1 AbbVie, 

Inc., North Chicago, IL; 2 Toronto Centre for Liver Disease, University 

of Toronto, Toronto, ON, Canada; 3 North Shore University 

Hospital, Manhasset, NY; 4 GastroOne, Germantown, TN; 5 Ghent 

University Hospital, Gent, Belgium; 6 Medical University of Vienna, 

Vienna, Austria 

Objective: Some individuals infected with HCV genotype 1 

treated with the 3 direct-acting antiviral (3D) regimen of ombitasvir 

(OBV), paritaprevir (PTV, identified by AbbVie and Enanta), 

and dasabuvir (DSV) require ribavirin (RBV) to optimize the 

chance of achieving a sustained virologic response (SVR). We 

describe the management of anemia-related adverse events 

(AEs) due to RBV-associated hemolysis in phase 3 trials for 

patients receiving the 3D + RBV regimen. Methods: Data were 

pooled from the SAPPHIRE-I and -II, PEARL-II, -III, and -IV, and 

TURQUOISE-II studies including patients with HCV genotype 

1a (N = 856) and 1b (N = 691) infection. Patients without cirrhosis 

received the 3D + RBV regimen for 12 weeks, whereas 

those with cirrhosis received 12 or 24 weeks treatment. Ribavirin 

was initially dosed according to body weight with a total 

daily dose of 1000 mg if


Ronald D’Amico - Employment: AbbVie; Stock Shareholder: AbbVie 




The following authors have nothing to disclose: Hans Van Vlierberghe, Greg Ball 

1068 

The healthcare cost burden of HCV-infected baby boomers 

in the US 

Ron Brookmeyer 2 , Timothy R. Juday 1 , Darius Lakdawalla 3,4 , Steven 

Marx 1 , Gigi Moreno 4 , Yuri Sanchez 1 ; 1 HEOR, AbbVie, Mettawa, 

ID; 2 David Geffen School of Medicine at UCLA, Los Angeles, CA; 

3 USC, Los Angeles, CA; 4 Precision Health Economics, Los Angeles, 

CA 

Background While the incidence of hepatitis-C virus (HCV) 

infection peaked more than 25 years ago, the untreated 

infected population is aging and experiencing expensive liver 

complications, raising questions about future financial burdens 

for Medicare. To shed light on this, we quantified healthcare 

costs for 50-64 year-old “baby boomers” with HCV by diagnosis 

and insurance status. Methods This study conducted a 

literature review and analyzed five waves of National Health 

and Nutrition Expenditures Survey (NHANES) data from 2003- 

2004 through 2011-2012. Insurance, HCV infection, and 

diagnosis status were obtained from NHANES. Respondents 

were identified as diagnosed if they tested positive for HCV, 

responded to the follow-up interview, and reported awareness 

of their HCV infection prior to NHANES. The analysis focused 

on respondents with HCV RNA, indicating chronic infection. 

The final sample contained 28,607 respondents. Healthcare 

costs for chronic HCV reported in the literature were applied 

to estimate total HCV costs. Results Among the 2.7 million 

Americans with chronic HCV, approximately 1 million are 

aged 50–64. Of these, about 319,003 are undiagnosed, 

and 161,609 are diagnosed but uninsured. Privately insured 

chronic HCV patients cost $17,176 annually if suffering from 

noncirrhotic disease (NCD), $22,752 if suffering from compensated 

cirrhosis (CC), and $59,995 if suffering from endstage 

liver disease (ESLD). This implies annual costs for the 

diagnosed, insured population of roughly $11.12 billion. If 

the diagnosed uninsured were treated like insured patients, 

this would add $3.23 billion. If the undiagnosed were diagnosed 

and treated like insured patients, this would add another 

$5.48 billion. Conclusions As baby boomers with HCV enter 

Medicare, the program may face substantial cost burdens. 

Proactively treating HCV patients before they suffer expensive 

long-term complications can help mitigate this burden to the US 

government. 

Gigi Moreno - Consulting: AbbVie 


1069 

An Actuarial View of the Aging Hepatitis C Virus (HCV) 

Epidemic 

Gabriela Dieguez 2 , Bruce Pyenson 2 , Ryan Cannon 2 , Steven 

Marx 1 , Yuri Sanchez 1 , Timothy R. Juday 1 ; 1 HEOR, AbbVie, Mettawa, 

ID; 2 Milliman, New York, NY 

PURPOSE In the coming years, the aging and treatment dynamics 

of people with chronic HCV infection may substantially shift 

the financial burden of health insurance coverage from private 

payers to Medicare. We examined shifts in the distribution of 

the HCV-infected population from 2015-2025 by US health 

insurance market, with a focus on people aging into Medicare 

and assuming that no patients would be cured of HCV infection. 

METHODS A stochastic actuarial model was developed to forecast 

the annual number of people with HCV insured by Medicare, 

Medicaid, commercial (including self-insured employers 

and Health Insurance Exchange enrollees), and military-associated 

insurance (including the Veteran Administration), as well 

as the number of uninsured HCV patients from 2015 to 2025. 

The current number of HCV patients in the major insurance 

markets and their annual probabilities of mortality, disability, 

and disease progression by age, gender, income, and HCV 

stage were estimated using data from Truven Health Market- 

Scan, Medicare 5% Sample, National Health and Nutrition 

Examination Survey, and Census Bureau. The expected impact 

of the Affordable Care Act on the HCV population’s health 

insurance status was considered. Prisoners were excluded from 

the analysis. Based on the HCV population forecast, the number 

of people with HCV who will gain Medicare eligibility 

through aging or disability in the next 10 years was calculated. 

RESULTS About 2.5 million non-institutionalized people in the 

US are estimated to be infected with HCV today. The majority 

of these patients are under age 65 with only 16% (405,000) 

covered by Medicare. This forecast predicts that, by 2025, 

38% (591,000) of people with HCV in the US are expected 

to be eligible for Medicare, and the number of HCV patients 

with commercial, Medicare, or military insurance is expected 

to decline to less than half of the current levels. In the next 10 

years, 635,000 more people with HCV will gain Medicare 

coverage. Furthermore, 921,000 people with HCV will die if 

left untreated. CONCLUSION Due to both age- and disease-related 

mortality, the HCV-infected population will decrease by 

almost 1 million over the next 10 years, assuming no treatment. 

By 2020, Medicare will displace commercial insurers as the 

largest payer for people who are currently infected with HCV. 

Sources: NHANES, Di Bisceglie, 2000, and Gordon 2012. 

Disclosures: 

Ron Brookmeyer - Consulting: Precision Health Economics 


Darius Lakdawalla - Consulting: AbbVie Pharmaceuticals; Stock Shareholder: 

Precision Health Economics 


Disclosures: 

Gabriela Dieguez - Consulting: Abbvie 

Bruce Pyenson - Consulting: AbbVie, which funded this research, Employed by 

Milliman, Inc., which consults to numerous payer, providers, and suppliers to the 

healthcare industry, PhRMA 

Ryan Cannon - Consulting: Abbvie, Milliman, Inc., PhRMA 



Timothy R. Juday - Employment: AbbVie; Stock Shareholder: AbbVie


1070 

Early and persistent increase of serum cholesterol and 

low-density lipoprotein in chronic hepatitis C patients 

during IFN-free treatment 

Clarissa Freissmuth, Albert Stättermayer, Karin Kozbial, Sandra 

Beinhardt, Rafael Stern, Michael Eder, Petra E. Steindl-Munda, 

Michael Trauner, Peter Ferenci, Harald Hofer; Department of Internal 

Medicine III, Division of Gastroenterology and Hepatology, 


Background&Aims: Hepatitis C virus (HCV) is closely related 

to lipid metabolism. HCV genotype-(GT)-3 infection is associated 

with hepatic steatosis and low levels of serum cholesterol, 

which are reversible after successful HCV treatment. Moreover, 

in HCV GT-1 infection serum levels of low-density lipoprotein 

(LDL) particles increase on HCV clearance during sofosbuvir/ 

ribavirin (SOF/RBV) therapy. This study investigated serum lipid 

parameters in patients with HCV-1 and 3 infection undergoing 

IFN-free treatment. Patients&Methods: Total serum cholesterol, 

high density lipoprotein (HDL), LDL and serum triglycerides (TG) 

were analyzed at baseline, during (initially weekly, afterwards 

every four weeks) and after antiviral therapy in 208 patients 

(GT-1: N=163, age: 57±11, BMI: 26±4kg/m 2 , male/female: 

99/64, F4: 108 (66%), Diabetes mellitus (DM): 35(21%)), 

(GT-3: N=45, age: 52±7, BMI: 26±4kg/m 2 , m/f: 32/13, F4: 

34 (76%), DM: 6 (13%)). Patients with GT-3 received SOF, 

400mg/d with RBV (N=24) or Daclatasvir (60mg/d) (DCV, 

N=21). Patients with GT-1 received SOF in combination with 

DCV (N=85), Simeprevir 150mg/d (N=56), 90mg/d Ledipasvir 

(N=20) or RBV (N=2). Results: Overall, serum cholesterol 

and LDL increased during treatment (table 1). This was 

observed after the first week of treatment (chol BL 

149±36 

vs. chol W1 

162±41 vs. chol W2 

169±40 mg/dL, mean±SD, 

p


results must be confirmed by analysis of the SVR12. Full result 

analysis will be presented at the congress. * Same contribution 

for the 2 first authors 

Disclosures: 


MSD, BMS 

Alexandre Pariente - Board Membership: Mayoli spindler; Speaking and Teaching: 

Janssen, Roche, BMS, Abbvie 

Thong Dao - Board Membership: Schering-Plough, Schering-Plough, Schering-Plough, 

Schering-Plough; Speaking and Teaching: Roche, Gilead, Roche, 

Gilead, Roche, Gilead, Roche, Gilead 

Alexandre Pariente - Board Membership: Mayoli spindler; Speaking and Teaching: 

Janssen, Roche, BMS, Abbvie 

Patrick Delasalle - Speaking and Teaching: ROCHE, AXCAN, BMS, GILEAD, 

MERCK, JANSSEN, ROCHE, AXCAN, BMS, GILEAD, MERCK, JANSSEN 

Xavier Causse - Board Membership: Gilead, Janssen-Cilag; Grant/Research 

Support: MSD; Speaking and Teaching: Gilead, BMS, Janssen-Cilag 

Francois Bourhis - Consulting: ABBVIE, GILEAD; Speaking and Teaching: BMS 


The following authors have nothing to disclose: Bruno Lesgourgues, Remy Andre, 

Brigitte Bernard-Chabert, Isabelle Rosa, Damien Lucidarme, Christophe Renou, 

Christine Silvain, Gilles Macaigne, Thierry Fontanges, Matthieu Schnee, Hortensia 

Lison, Christophe Pilette, Jean Pierre Arput, Frédéric Heluwaert 

1073 

Safety And Efficacy Of The Combination Simeprevir-Sofosbuvir 

In HCV Genotype 1- And 4-Mono-Infected 

Patients From The French Observational Cohort Anrs 

Co22 Hepather* 

ANRS/AFEF HEPATHER CO22 study group; National Agency for 

Research on AIDS and Viral Hepatitis, Paris, France 

Background and aims. Real-life results of the Sofosbuvir/Simeprevir 

combination have been extensively reported from USA 

but there are few data from other geographical areas. In May 

2015, more than 3500 patients of the French observational 

cohort ANRS CO22 HEPATHER were given the new oral antivirals 

in 32 centers: we report the preliminary results of the Sofosbuvir/Simeprevir 

combination in Genotype 1- and 4-infected 

patients. Methods. Demographics, history of liver disease 

were collected at entry in the cohort. Clinical, adverse events, 

and virological data were collected throughout treatment and 

post-treatment follow-up. Results. 552 HCV (433 genotype 1- 

and 119 genotype 4-) mono-infected patients were given a 

combination of Sofosbuvir (SOF: 400 mg/d) and Simeprevir 

(SMV: 150 mg/d) without Ribavirin (n=491) or with Ribavirin 

(1-1.2 g/d, n=61). 54% of patients had cirrhosis (3% decompensated), 

75% were previously treated with PR (n=315) or 

PR + a first generation PI (n=25). Overall, the SVR12 rate was 

88%. Underlying cirrhosis did not impact SVR rates. In patients 

who received the 12-week combination SOF/SMV +/- RBV the 

SVR12 rate differed significantly according to the genotype 

(80% in GT1a, 92% in GT1b and 86% in GT4, p=0.004) and 

according to the biochemical markers of liver severity -MELD 

score, total and conjugated bilirubin, platelet count, Prothrombin 

time, ALT, AST-, p50 gms/day), active cocaine or crack use Patient characteristics- 

table to follow on the slide/poster They were divided into 

2 groups; Group A (n=14): SOF 400 mg + LDV 90 mg (daily 

once) - 4 weeks Group B (n=15): SOF 400 mg + SIM 150 mg 

(daily once) - 8 weeks Labs: Prior to therapy- HCV RNA 0 and 

3 weeks, sickle cell panel, LFT’s, CBC, SMA On therapy- HCV 

RNA on day 0, 1 week, 4 week, 6 week followed by 16 th and 

20 th week (SVR), LFT’s, CBC, SMA Results: Table Conclusion: 

This study demonstrates a high SVR with short-course DAA’s in 

acute hepatitis C with SVR (at 20 weeks) >90% in both groups. 

The drugs were well tolerated. 

Results 

Disclosures: 


John, M. Aloysius, Robert Brown


1075 

Sofosbuvir, Ledipasvir in IBD treated patients with 

advanced biologics including Ribavirin eradicating 

Chronic Hepatitis C: SOLATAIRE C Trial. A multi-center 

clinical prospective pilot study 

Patrick Basu 1,2 , Niraj J. Shah 3 , Nimy John 2 , M. Aloysius 2 , Robert 

Brown 4 ; 1 Columbia University School of Physicians and Surgeons, 

Forest hills, NY; 2 King’s County Hospital Medical Center, NY, 

New York, NY; 3 James J. Peters VA Medical Center, Icahn School 

of Medicine at Mount Sinai, NY, New York, NY; 4 Weill Cornell 

Medical College, New York, NY 

Background: The prevalence and concomitant treatment of 

chronic HCV and IBD has not been elucidated. IBD and CHC 

of >20 years duration may have more fibrosis due to immune 

suppression. Treatment of IBD often requires biological therapies 

to achieve longer disease-free remission, which further 

accelerates higher HCV replication. Thus effective therapy for 

IBD patients requiring immunosuppressive therapy is needed 

Aim: To evaluate the role and efficacy of new NS5A + NS5B 

inhibitors with and without RBV in treating CHC in moderate 

to severe IBD requiring biologic therapy Methods: 35 patients 

were recruited from the Kings County Hospital Medical Center, 

Brooklyn. Inclusion criteria CHC with IBD, Age: >18, HCV Viral 

Load: > 400,000IU/mL, Genotype 1. Fibrotic Score: Metavir 

F1 to F4 Primary End Point: SVR12 Secondary End Point: IBD 

activity index, sustained control of symptoms SVR 24 and complete 

IBD remission at 30 weeks (endoscopy at 30 weeks) 

The patients were divided into two groups: Group A (n=17)- 

RBV 1000 mg + LDV and SOF; for 8 weeks Group B (n=18): 

LDV and SOF; for 12 weeks All will have base line RAV and 

ETRAV’s and SVR 24. RAVs 5A polymorphism by Quest. Viral 

loads 0, 7, 14 days; 4 th , 8 th , 12 th and 24 th weeks Patient characteristics: 

list to follow in the poster/oral presentation Results: 

See table. 24 th week RAV- results are pending Conclusion: This 

study demonstrates that DAA’s for HCV appear to have similar 

efficacy and safety in the presence of active IBD therapy with- 

TNF Alfa antagonists while keeping the IBD in uninterrupted 

remission 

Results 

Disclosures: 


John, M. Aloysius, Robert Brown 

1076 

Sofosbuvir plus Ribavirin in the Treatment of Egyptian 

Patients with Chronic Genotype 4 HCV Infection 

Wahid H. Doss 1 , Peter J. Ruane 2 , Gamal Shiha 3 , Dani Ain 2 , 

Reham Soliman 3 , Marwa Khairy 4 , Mohamad Hassany 1 , Joseph 

Riad 2 , Waleed Samir 3 , Rabab F. Omar 4 , Radi Hammad 1 , Raymond 

Meshrekey 2 , Mostafa Gamil 4 , Deyuan Jiang 5 , Benedetta 

Massetto 5 , Steven J. Knox 5 , Kathryn Kersey 5 , John G. McHutchison 

5 , Gamal E. Esmat 4 ; 1 National Hepatology and Tropical Medicine 

Research Institute, Cairo, Egypt; 2 Ruane Medical and Liver 

Health Institute, Los Angeles, CA; 3 Egyptian Liver Research Institute 

and Hospital, Mansoura, Egypt; 4 University of Cairo, Cairo, 

Egypt; 5 Gilead Sciences, Inc., Foster City, CA 

Background: HCV infection is a major public health burden in 

Egypt, with an estimated 6 million people chronically infected. 

The combined safety and efficacy data for sofosbuvir (SOF) 

plus ribavirin (RBV) from 2 studies in Egyptian patients with 

chronic genotype (GT) 4 HCV infection are summarized here. 

Methods: An integrated analysis was conducted of data from 

treatment-naïve and treatment-experienced patients enrolled in 

Study GS-US-334-0114 in the USA (n=60) and Study GS-US- 

334-0138 in Egypt (n=103). In each study, subjects were randomized 

1:1 to receive either 12 or 24 weeks of SOF (400 

mg daily) + RBV (1000-1200 mg daily). Exact logistic regression 

analyses were used to explore the relationships between 

SVR12 rates and baseline characteristics in the combined dataset. 

Results: Overall, 67% of patients were male, mean age 

(range) was 49 years (19-75), 42% had BMI ≥30 kg/m 2 , 19% 

had cirrhosis, 82% had IL28B non-CC genotype, 56% had 

HCV RNA ≥800,000 IU/mL at baseline, and 53% were treatment-experienced. 

SVR12 rates were 73% (61/83) and 91% 

(73/80) for SOF+RBV administered for 12 and 24 weeks, 

respectively. Multivariate logistic regression analysis identified 

male sex, IL28B non-CC genotype, and baseline HCV RNA 

≥800,000 IU/mL as statistically significant factors associated 

with worse treatment outcome while 24 weeks of treatment 

increased the likelihood of achieving SVR12. In treatment-naïve 

patients without cirrhosis, SOF+RBV for 12 or 24 weeks 

resulted in SVR12 rates of 88% (29/33) and 94% (30/32), 

respectively. The most common adverse events were headache, 

fatigue, insomnia, cough and pruritus. Most AEs were mild or 

moderate in severity and none resulted in treatment discontinuation; 

5 patients experienced SAEs of which 1 (dyspnea) 

was considered treatment-related. Conclusions: Integrated data 

from these 2 studies show SOF+RBV was well tolerated with 

an AE profile consistent with that of RBV. SOF+RBV for 24 

weeks resulted in a 91% SVR12 rate in Egyptian patients with 

HCV GT4 infection. The combined safety and efficacy data 

support the use of this interferon-free regimen. In addition, the 

data suggest a shorter treatment duration of 12 weeks may be 

of benefit for some patients, such as treatment naïve patients 

without cirrhosis. 

Disclosures: 






Radi Hammad - Grant/Research Support: Gilead Sciences,Inc, Janssen Pharmaceuticals, 

Inc. 

Deyuan Jiang - Employment: Gilead Sciences 

Benedetta Massetto - Employment: Gilead Sciences, Inc.; Stock Shareholder: 

Gilead Sciences, Inc 


Kathryn Kersey - Employment: Gilead Sciences, Inc; Stock Shareholder: Gilead 

Sciences, Inc



Sciences 

Gamal E. Esmat - Advisory Committees or Review Panels: MSD &BMS companies, 

MSD &BMS companies, Abbvie; Grant/Research Support: Gilead Sc; 

Speaking and Teaching: Roche & GSK companies, Roche & GSK companies 

The following authors have nothing to disclose: Wahid H. Doss, Gamal Shiha, 

Dani Ain, Reham Soliman, Marwa Khairy, Joseph Riad, Waleed Samir, Rabab 

F. Omar, Raymond Meshrekey, Mostafa Gamil 

1077 

The TOSCAR Study: Sofosbuvir and Daclatasvir Therapy 

for Decompensated HCV Cirrhosis with MELD scores ≥ 

15: What is the point of no return? 

Geoff McCaughan 1 , Stuart K. Roberts 2 , Simone I. Strasser 1 , Paul 

Gow 3 , Alan J. Wigg 4 , Caroline Tallis 5 , Gary P. Jeffrey 6 , Jacob 

George 7 , Alex J. Thompson 8 , Susan Mason 9 , Joanne Mitchell 2 , 

Peter W. Angus 3 ; 1 ANLTU, Royal Prince Alfred Hospital/Univeristy 

of Sydney, Sydney, NSW, Australia; 2 Alfred Health, Melbourne, 

VIC, Australia; 3 VLTU, Melbourne, VIC, Australia; 4 SALTU, Adelaide, 

SA, Australia; 5 QLTU, Brisbane, QLD, Australia; 6 WALTU, 

Perth, WA, Australia; 7 Westmead Hospital, Sydney, NSW, Australia; 

8 St Vincent’s Hospital, Melbourne, VIC, Australia; 9 AW Morrow 

Gastroenterology & Liver Centre, Royal Prince Alfred Hospital, 

Sydney, NSW, Australia 

Background: Interferon free therapies for chronic HCV have 

resulted in greater than 95% SVRs. However, there are few 

data that examine the efficacy in patients with decompensated 

chronic liver disease due to HCV. We report such data in a 

well defined patient population. Aims: To examine virological 

responses in patients with decompensated HCV liver disease 

treated with an interferon free regime To examine the effect of 

viral clearance on severity and outcomes of liver disease. Methods: 

A compassionate access Australia-wide protocol using 

Sofosbuvir and Daclatasvir for 24 weeks without Ribavirin to 

treat patients with HCV decompensated liver disease with a 

MELD score of ≥ 15 was introduced in November 2014. Data 

was collected from multiple sites at tertiary level liver centres 

throughout Australia. Results: 92 patients have been analysed. 

The average age was 55 years. 73% were male. 39% had 

genotype 1a, 9% genotype1b and 27% genotype 3. The average 

Child Pugh Score (CPS) was 9.5 with an average MELD 

of 17.4. 23 patients (25%) had MELD scores > 20 (maximum 

29). 76% of patients had ascites and 65% had encephalopathy. 

35 patients were listed and 10 underwent liver transplantation 

during this period .6 patients died before transplantation 

(5 within 8 weeks of starting treatment). End of treatment (EOT) 

data was available in 15 patients. All but 1 patient was HCV 

PCR negative. The single patient who remained positive had 

undergone a surgical procedure with treatment disruption. This 

patient had also had a previous relapse from a 12 week course 

of Sofosbuvir and Ledipasvir. 8 of 15 patients (approx 50%) 

had a ≥ 2 improvement in MELD scores whilst 4 (approx 25%) 

had an increase in MELD score of ≥ 2. Improvement in MELD 

≥ 2 was only seen in patients with a pre-treatment MELD of < 

20. The improvements in CPS ≥ 2 were seen in 6/15 (40%). 

SVR12 data and post-transplant HCV RNA status for all patients 

will be presented. Conclusion: Sofosbuvir/Daclatasvir therapy 

in patients with significantly decompensated HCV liver disease 

results in on-treatment virological control in almost all patients. 

Despite this, EOT improvements in MELD scores were only seen 

in 50% of patients. Improvements were limited to patients with 

MELD scores < 20. SVRs and further assessment of liver disease 

severity are currently being analysed. 

Disclosures: 



Simone I. Strasser - Advisory Committees or Review Panels: Janssen, AbbVie, 

Roche Products Australia, MSD, Bristol-Myers Squibb, Gilead, Norgine, Bayer 

Healthcare; Speaking and Teaching: Bayer Healthcare, Bristol-Myers Squibb, 

MSD, Roche Products Australia, Gilead, Janssen, Abbvie 

Jacob George - Advisory Committees or Review Panels: Roche, BMS, MSD, 

Gilead, Janssen, Abbvie; Grant/Research Support: MSD 




Abbvie, BMS 

Susan Mason - Advisory Committees or Review Panels: Janssen, MSD, BMS; 

Grant/Research Support: Janssen, MSD; Speaking and Teaching: ASHM 



The following authors have nothing to disclose: Paul Gow, Alan J. Wigg, Caroline 

Tallis, Gary P. Jeffrey, Joanne Mitchell 

1078 

(THE SOFGER TRIAL) Sofosbuvir-based treatment under 

real life conditions in Germany 

Peter Buggisch 1 , Christoph Sarrazin 2 , Stefan Mauss 3 , Holger 

Hinrichsen 4 , Karl-Georg Simon 5 , Dietrich Hueppe 6 , Johannes Vermehren 

2 , Barbara Seegers 4 , Joerg Petersen 1 ; 1 IFI, Liver Center 

Hamburg, Hamburg, Germany; 2 1 Medizinische Klinik, Johann 

Wolfgang Goethe University Hospital, Frankfurt, Germany; 3 Zentrum 

für HIV und Hepatogastroenterologie, Duesseldorf, Germany; 

4 Gastroenterolgische Praxis, Kiel, Germany; 5 Gastroenterologisch-hepatologisches 

Zentrum, Leverkusen, Germany; 6 Gastroenterologische 

Gemeinschaftspraxis, Herne, Germany 

Background and Aims: After the approval of direct-acting antivirals 

(DAA) in Germany 2014 starting with Sofosbuvir(SOF) 

Jan., Simeprevir(SIM) May, Daclatasvir(DCV) August and Ledipasvir(LDV) 

November, recommendations in Germany (BNG/ 

DGVS) were rapidly changed accordingly. Rules for reimbursement 

changed over time,too, but there was no limitation to 

advanced fibrosis stages. Aim of this study was to evaluate 

the change of treatment modalities following availability of 

treatment options, implementation of recommendation/reimbursement 

rules as well as the safety and efficacy of the SOF 

based therapies under real life conditions. Methods: In this 

non-interventional, prospective, multi-center study conducted 

by the Association of German Gastroenterologists in private 

practice (BNG) (5 center) and one academic based center 

(Frankfurt), SOF-based therapies from Jan.2014 to February 

2015 are being evaluated in Germany. Demographic, clinical, 

virology data and adverse events are collected throughout 

treatment and post-treatment. This interim analysis shows data 

about pts.with SOF-based treatments. Results: 790 patients 

(471 male, 318 female) received SOF-based treatments. 553 

pts.with genotype 1(275 1a, 278 1b), 61 genotype 2, 128 

genotype 3, 48 genotype 4. 393 (49,8% had previous treatment 

(65 with DAAs), 417 of pat.had fibrosis F3-4. In 118 pat. 

no exact fibosis stage was evalueted, but 45% of those were 

clinically cirrhotic. 93 pat. received SOF/Riba (84% Genotype 

2(12 w) and 3(24 w) 160 received PEG/RIBA/SOF 12 w 

(all before Sep.2014) 118 SIM/SOF, 221 SOF+DCV ±Riba 

(75% F3/4, 198 SOF/LDV (58% F0-2). No IFN-based treatment 

started after 9/2014. SVR 4 for SIM/SOF (116 pat.) is 

87% SVR 12 (90 pat.) 86 %. SVR 4 for PEG/RIBA/SOF (157 

pat.) is 83%, SVR 12 (144 pat.)79%. SVR 4 for SOF/RIBA is 

78%(88 pat.) SVR 12 75% (82 pat). SVR 4 for SOF/DCV is 

85% (163 pat.) SVR 12 (97 pat.) 87%. SVR 4 for SOF/LDV 

is 96% (110 pat.). There was a clear trend to use SOF/LDV 

in lower fibrosis stages (8 or 12 weeks). Anemia (Riba-based 

therapy),headache, sleeping disorders and fatigue were the 

most reported AEs. No severe side effects occurred. 6 deaths 

were reported due to complications of cirrhosis. More SVR


12 data and detailed results for the different treatments will 

be available at the meeting. Conclusions: SOF-based triple 

therapies under real life conditions seems to have comparable 

results to clinical trials and other real life cohorts.There was a 

fast switch to IFN-free therapies in Germany following the recommendations. 

Advanced patients were treated first, but now 

almost 40% were treated without advanced fibrosis. Adverse 

events and treatment discontinuations were low. 

Disclosures: 

Peter Buggisch - Advisory Committees or Review Panels: Janssen, AbbVie, BMS; 

Speaking and Teaching: Roche, MSD, Gilead, Merz Pharma 









Holger Hinrichsen - Advisory Committees or Review Panels: BMS, Janssen, Gilead, 

Abbvie; Speaking and Teaching: MSD 

Karl-Georg Simon - Advisory Committees or Review Panels: AbbVie, BMS, 

JANSSEN; Speaking and Teaching: AbbVie, BMS, FALK, GILEAD, JANSSEN, 

NORGINE 



Johannes Vermehren - Advisory Committees or Review Panels: Abbott; Speaking 

and Teaching: BMS, Covidien 





The following authors have nothing to disclose: Barbara Seegers 

1079 

Quality-Adjusted Cost of Care for Treatment Naive (TN) 

Patients with Genotype 1 (GT1) Chronic Hepatitis C (CH- 

C): An Assessment of Innovation Cost of Drug Regimens 

versus the Value of Health Gains to the Society 

Zobair M. Younossi 1,2 , Haesuk Park 3 , Douglas Dieterich 4 , Sammy 

Saab 5 , Aijaz Ahmed 6 , Stuart C. Gordon 7 ; 1 Center For Liver 

Disease, Department of Medicine, Inova Fairfax Hospital, Falls 

Church, VA; 2 Betty and Guy Beatty Center for Integrated Research, 

Inova Health System, Falls Church, VA; 3 University of Florida, 

Gainesville, FL; 4 Mount Sinai Medical Center, New York City, NY; 

5 University of California Los Angeles, Los Angeles, CA; 6 Stanford 

University, Standford, CA; 7 Henry Ford Hospital, Detroit, MI 

Objectives: New HCV regimens have increased cure rates and 

treatment costs. Quality-adjusted cost of care (QACC) provides 

an approach for assessing cost growth of regimens in the 

context of value of health [quality-adjusted life-years (QALY)] 

improvements delivered by these regimens (Lakdawalla, Health 

Affairs 2015). Our aim was to assess QACC for regimens 

approved for GT1 CH-C in the US. Methods: A decision analytic 

Markov compared treatment of TN GT1 CHC patients 

with approved regimens including pegylated interferon and 

ribavirin (PR), 1st generation triple (boceprevir+PR and telaprevir+PR), 

2nd generation triple (sofosbuvir+PR and simeprevir+PR) 

and all-oral regimens (ledipasvir/sofosbuvir and 

ombitasvir+paritaprevir/ritonavir+dasabuvir±ribavirin). Sustained 

virologic response (SVR), transition probabilities, utility, 

and mortality were based on literature review and consensus 

by hepatologists. Drug costs were from Redbook using wholesale 

acquisition costs. QACC was defined as the increase in 

treatment price minus the increase in the patient’s QALYs multiplied 

by the value of a QALY ($50,000). Since this threshold 

may be outdated, we also estimated the value of a QALY to 

be between $100,000 and $300,000. Results: Average drug 

costs increased by $41,311, $54,249, and $52,352 for 1st, 

2nd generation triples and all-oral therapies compared to PR 

($35,331). Health improvements were 0.121, 0.692, and 

1.366 QALYs, valued at $6,057, $34,622, and $68,318 

for 1st, 2nd generation triples and all-oral therapies compared 

to PR. Compared to PR, 1st and 2nd generation triples could 

cost the society $35,254 and $19,627. On the other hand, 

all-oral therapy could save the society $15,966. By increasing 

the value threshold for a QALY up to $300,000, all-oral regimens 

save the society up to $357,555. Conclusions: Despite 

the higher cost of all-oral regimens, this QACC analysis shows 

substantial value to the society. 

Disclosures: 



Haesuk Park - Consulting: Gilead Science 



Sammy Saab - Advisory Committees or Review Panels: BMS, Gilead, Merck, 

Janssen; Grant/Research Support: Gilead; Speaking and Teaching: BMS, Gilead, 

Merck, Janssen, Salix, Onyx, Bayer, Janssen; Stock Shareholder: Achillion, 

Johnson and Johnson, BMS, Gilead 




Inc. 




1080 

Asian Patients with Chronic Genotype 2 HCV Infection 

Achieve 98% Sustained Virologic Response Following 

12 Week administration of Sofosbuvir in Combination 

with Ribavirin: Integrated analysis of Phase 3 Multicenter 

Studies 

Masao Omata 1 , Young-Suk Lim 2 , Wan-Long Chuang 3 , Jia-Horng 

Kao 4 , Bing Gao 5 , Shampa De-Oertel 5 , Jenny C. Yang 5 , Hongmei 

Mo 5 , Diana M. Brainard 5 , Steven J. Knox 5 , John G. McHutchison 

5 , Masashi Mizokami 6 , Kwang-Hyub Han 7 ; 1 Yamanashi Prefectural 

Hospital Organization, Yamanashi, Japan; 2 Asan Medical 

Center, University of Ulsan College of Medicine, Seoul, Korea 

(the Republic of); 3 Kaohsiung Medical University, Kaohsiung, Taiwan; 

4 National Taiwan University College of Medicine, Taipei, 

Taiwan; 5 Gilead Sciences, Inc, Foster City, CA; 6 National Center 

for Global Health and Medicine, Tokyo, Japan; 7 Yonsei University 

College of Medicine, Seoul, Korea (the Republic of) 

Introduction: Chronic hepatitis C (CHC) infection presents a 

significant burden on public health in Asia. Among patients


with CHC, 20-30% in Japan and 50% in Korea and Taiwan 

are infected with hepatitis C virus (HCV) genotype (GT) 2. 

Sofosbuvir (SOF), an NS5B polymerase inhibitor, in combination 

with ribavirin (RBV) is the first all-oral regimen for 

treatment of HCV GT2 infection. The aim of this integrated 

analysis is to characterize the efficacy and safety of SOF+RBV 

in a large cohort of Asian patients with HCV GT2 infection. 

Methods: This integrated analysis combines results from two 

Phase 3 trials, GS-US-334-0118 (Japan) and GS-US-334- 

0115 (Korea and Taiwan). In both studies, treatment-naïve 

and treatment-experienced adults with chronic GT2 HCV infection 

received SOF (400mg) combined with RBV (weight-based 

dosing) for 12 weeks. Eligibility criteria: no upper age limit 

restriction, inclusion of subjects with compensated cirrhosis, 

platelet count ≥50,000/mL and no restriction on neutrophil 

count. The primary efficacy endpoint was Sustained Virologic 

Response measured 12 weeks after the last dose of study drug 

(SVR12). Results: Overall, 369 patients were enrolled (n=129 

Korea, n=87 Taiwan, and n=153 Japan), of which 64% 

(238/369) were treatment naïve and 36% (131/369) were 

treatment experienced. Mean age (range) was 55 (22-82) 

years, 44% male (164/369), 82.1% (303/369) IL28B-CC, 

and 12% (43/369) had cirrhosis. The overall SVR12 rate was 

98% (360/369). Similar high rates of SVR12 were seen in 

treatment-experienced (98%, 128/131), patients ≥65 years 

old (96%, 73/76), and those with cirrhosis (98%, 42/43). 

Among patients who did not achieve SVR12 (n=9), 1 was a 

partial responder, 6 relapsed, and 2 were lost to follow up. 

Adverse events were generally mild to moderate. The most 

common adverse events were nasopharyngitis (17%, 63/369), 

headache (12%, 45/369), and pruritis (11%, 41/369). Laboratory 

abnormalities were infrequent and consistent with the 

safety profile of RBV. No AEs led to treatment discontinuation. 

Conclusions: Treatment-naïve and treatment-experienced Asian 

patients in Japan, Korea, and Taiwan with chronic GT2 HCV 

infection, including those with compensated cirrhosis, achieved 

high rates of SVR12 with 12 weeks of an IFN-free, all-oral 

regimen of SOF+RBV. The regimen was safe and well-tolerated 

with no treatment discontinuations due to AE and the overall 

AE profile was consistent with that observed with RBV. The 

data suggest that SOF+RBV may offer an improved, IFN-free 

treatment for Asian patients with chronic GT2 HCV infection. 

Disclosures: 

Masao Omata - Advisory Committees or Review Panels: Boehringer Ingelheim; 

Speaking and Teaching: Otsuka Pharmaceutical, Bayer 



Novartis 



Bing Gao - Employment: Gilead; Stock Shareholder: Gilead 

Shampa De-Oertel - Employment: Gilead Sciences, Inc; Stock Shareholder: Gilead 

Sciences, Inc 




Sciences 



Sciences 

The following authors have nothing to disclose: Jia-Horng Kao, Masashi 

Mizokami, Kwang-Hyub Han 

1081 

Sofosbuvir and ledipasvir for 8 weeks in patients with 

hepatitis C virus (HCV) mono-infection and human 

immunodeficiency virus (HIV)-HCV co-infection with 

genotype 1 and 4 in clinical practice – Results from the 

GErman hepatitis C COhort (GECCO) 

Stefan Christensen 1 , Stefan Mauss 2 , Dietrich Hueppe 3 , Knud 

Schewe 4 , Thomas Lutz 5 , Jürgen K. Rockstroh 6 , Marcel Schuetze 7 , 

Guenther Schmutz 2 , Karl-Georg Simon 8 , Heiner W. Busch 1 , Patrick 

Ingiliz 7 , Axel Baumgarten 7 ; 1 Center for interdisciplinary Medicine 

(CIM), Muenster, Germany; 2 Center for HIV and Hepatogastroenterology, 

Duesseldorf, Germany; 3 Practice for Gastroenterology, 

Herne, Germany; 4 Infektionsmedizinisches Centrum Hamburg, 

Hamburg, Germany; 5 Infektiologikum, Frankfurt, Germany; 6 Internal 

Medicine I, University of Bonn, Bonn, Germany; 7 Medizinisches 

Infektiologie Zentrum Berlin, Berlin, Germany; 8 Practice 

for Gastroenterology Leverkusen, Leverkusen, Germany 

Introduction The first direct acting antivirals (DAA) were 

approved in Europe in 2014 based on limited study data. In 

particular, sofosbuvir (SOF) and ledipasvir (LDV) for 8 weeks in 

HCV-mono and HIV co-infected patients had not been systematically 

studied. Here, we present real-life data on efficacy and 

safety of SOF/ LDV for 8 weeks from Germany. Methods In this 

multicenter cohort, all patients who were started on sofosbuvir-based 

treatments were documented. For the current analysis, 

only patients with HCV genotype (GT) 1 and 4 on SOF/LDV for 

8 weeks were included. All patients within the GECCO cohort 

are part of the prospective German hepatitis C registry. Results 

Until May 2015, 836 patients on SOF-containing regimens 

have been enrolled. Of those, 629 (75%) are HCV-monoinfected 

and 207 (25%) HIV-HCV co-infected. 95/836 (92 GT 1 

and 3 GT 4) patients were treated with SOF/LDV for 8 weeks. 

19/95 patients (19%) had been unsuccessfully treated before, 

mainly with dual therapy consisting of pegylated interferon and 

ribavirin. 16/95 (17%) were HIV coinfected, 14 of those with a 

HIV-RNA < 15 cop/ml. 40/95 (42%) were male, median age 

(IQR) was 53 years (23-81). Median HCV-RNA was 809.900 

IU/ml (IQR 5.000-14.100.000). 6/95 (6%) had a HCV-RNA 

at baseline of > 6 mio IU/ml. 59/95 (62%) had liver stiffness 

measurement by transient elastography (Fibroscan®), with a 

median of 6,1kPa. Liver cirrhosis defined by a value above 

12.5kPa was present in 2 cases. 44/95 have reached post 

treatment week 4. All (44/44) had a negative HCV-RNA at 

post treatment week 4 (SVR 4). No premature discontinuations 

or lost to follow up are documented. Conclusion: In this this 

real life patient population with mainly low HCV viral load and 

without significant fibrosis we found high response rates in GT 

1 and GT 4 patients treated with SOF/LDV for 8 weeks. Even 

in pretreated patients, patients with advanced fibrosis and in 

patients with a HCV-RNA > 6 mio IU/ml, all not candidates 

for a short course SOF/LDV according to the German HCV 

treatment guidelines, 8 weeks of SOF/LDV seems to be safe 

and effective. 

Disclosures: 








Knud Schewe - Advisory Committees or Review Panels: abbvie, gilead, msd, 

bms, janssen cilag, viiv, hexal 

Thomas Lutz - Advisory Committees or Review Panels: Gilead, MSD, AbbVie, 

BMS, Janssen Cilag; Grant/Research Support: Gilead, GlaxoSmithKline, Roche, 

MSD, AbbVie, Janssen Cilag; Speaking and Teaching: ViiV, BMS






Karl-Georg Simon - Advisory Committees or Review Panels: AbbVie, BMS, 

JANSSEN; Speaking and Teaching: AbbVie, BMS, FALK, GILEAD, JANSSEN, 

NORGINE 

Heiner W. Busch - Speaking and Teaching: Tibotec, MSD, Janssen, BMS, Gilead 

The following authors have nothing to disclose: Marcel Schuetze, Guenther 

Schmutz, Patrick Ingiliz, Axel Baumgarten 

1082 

Hepatitis C virus NS5A L31V plus Y93H variants emerging 

after treatment failure with daclatasvir/asunaprevir 

are resistant to ledipasvir and sofosbuvir-like nucleotide 

polymerase inhibitor GS-558093 in human hepatocyte 

chimeric mice 

Yugo Kai, Hayato Hikita, Tomohide Tatsumi, Kazuhiro Murai, Yuto 

Shiode, Yasutoshi Nozaki, Yuki Makino, Tasuku Nakabori, Yoshinobu 

Saito, Naoki Morishita, Satoshi Tanaka, Ryotaro Sakamori, 

Naoki Hiramatsu, Tetsuo Takehara; Department of Gastroenterology 

and Hepatology, Osaka University Graduate School of Medicine, 

Suita, Japan 

Background: Resistance-associated variants (RAVs) emerge 

in hepatitis C virus (HCV) infection upon treatment failure of 

direct-acting antivirals (DAAs). Treatment failure of the asunaprevir 

(ASV) and daclatasvir (DCV) regimen is associated 

with emergence of RAVs at both L31 and Y93 in the HCV 

NS5A region. In the present study, we used human hepatocyte 

chimeric mice to examine the therapeutic effect of ledipasvir 

(LDV) and GS-558093 (NS5B NI), a nucleotide NS5B polymerase 

inhibitor like sofosbuvir, as a re-treatment option for 

L31 and Y93 treatment emergent RAVs. Methods: Thymidine 

kinase transgenic NOG (TK-NOG) mice with humanized liver 

were inoculated with serum from treatment-naïve and treatment-experienced 

patients with chronic hepatitis C under the 

approval of the institutional ethics committee. After developing 

persistent HCV infection, mice were administrated DAAs orally. 

HCV RNA levels were measured by TaqMan PCR, and population 

and deep sequencing were performed. Results: Human 

hepatocyte chimeric mice were inoculated with serum from 

either a patient who had failed to achieve sustained virologic 

response (SVR) by the ASV/DCV therapy or from a patient who 

had not experienced DAA therapy. After inoculation, the mice 

developed persistent HCV infection with HCV RNA levels up to 

5-7 LC/ml in serum. Population and deep sequencing revealed 

that the formers carried NS3/4A D168V, NS5A L31V plus 

Y93H triple mutant virus while the latter carried wild-type virus 

at these positions. Mice in each group were divided into 2 

treatment groups: one received 4 weeks of ASV/DCV administration 

and the other received 4 weeks of LDV/ NS5B NI. 

Serum HCV RNA levels in mice with wild-type HCV rapidly 

declined and sustained undetectable levels after ASV/DCV 

or LDV/ NS5B NI. In contrast, serum HCV RNA levels in mice 

with mutant virus reduced 1 to 2 LC/mL or 2 to 3 LC/mL from 

baseline by ASV/DCV or LDV/ NS5B NI therapy, respectively, 

but did not achieve undetectable levels at the end-of-treatment. 

Although mice infected with mutant virus failed to eradicate 

HCV by both therapies, no additional substitutions including 

NS5B S282T were detected. These mice were re-treated by 

combined administration of NS5B NI and telaprevir (TVR), of 

which resistance profile differs from ASV. Serum HCV RNA 

levels rapidly reached to undetectable levels. Discussion/Conclusion: 

The triple mutant virus emerging upon the treatment 

failure of ASV and DCV is relatively resistant to LDV/ NS5B 

NI. Other DAA regimens that do not share cross-resistance 

at NS3/4A D168, NS5A L31 and Y93 positions may be a 

choice of re-treatment for such case. 

Disclosures: 






The following authors have nothing to disclose: Yugo Kai, Tomohide Tatsumi, 

Kazuhiro Murai, Yuto Shiode, Yasutoshi Nozaki, Yuki Makino, Tasuku Nakabori, 

Yoshinobu Saito, Naoki Morishita, Satoshi Tanaka, Ryotaro Sakamori, Naoki 

Hiramatsu 

1083 

Sofosbuvir and Ribavirin for Six Weeks Is Not Effective 

Among People with Acute and Recently Acquired HCV 

Infection: The DARE-C II Study 

Marianne Martinello 1 , Edward J. Gane 2 , Margaret Hellard 3 , Joe 

Sasadeusz 4 , David Shaw 5 , Kathy Petoumenos 1 , Tanya L. Applegate 

1 , Jason Grebely 1 , Laurence Maire 1 , Philippa Marks 1 , David 

Cooper 1 , Gregory Dore 1 , Gail Matthews 1 ; 1 Kirby Institute, UNSW 

Australia, Sydney, NSW, Australia; 2 Auckland Hospital, Auckland, 

New Zealand; 3 Burnet Institute, Melbourne, VIC, Australia; 

4 Royal Melbourne Hospital, Melbourne, VIC, Australia; 5 Royal 

Adelaide Hospital, Adelaide, SA, Australia 

Background: Sofosbuvir (SOF) and ribavirin (RBV) are effective 

and well tolerated when administered for 12–24 weeks in genotype 

(GT) 1-4 chronic hepatitis C virus (HCV) infection. The 

aim of this study was to assess the efficacy of SOF and RBV for 

six weeks in individuals with acute or recently acquired HCV 

infection. Methods: In this multicentre study conducted in Australia 

and New Zealand, adults with recent HCV (duration of 

infection


ever, efficacy was suboptimal. Further research is needed to 

determine whether more potent DAA regimens with or without 

RBV will allow treatment duration to be shortened in acute or 

recently acquired HCV infection. 

Disclosures: 




Joe Sasadeusz - Grant/Research Support: Gilead Sciences, BMS, Roche, Janssen; 

Speaking and Teaching: Gilead Sciences, Roche, BMS 

Kathy Petoumenos - Grant/Research Support: Gilead Sciences 






Gail Matthews - Advisory Committees or Review Panels: gilead; Consulting: Viiv; 

Grant/Research Support: Gilead Sciences, janssen; Speaking and Teaching: 

BMS, MSD 

The following authors have nothing to disclose: Marianne Martinello, Margaret 

Hellard, David Shaw, Tanya L. Applegate, Laurence Maire, Philippa Marks, 

David Cooper 

patients, including 6 with GT1b infection, received 3D+RBV 

for 24 weeks, and 13 GT1b patients received treatment without 

RBV for 24 weeks. SVR4 was achieved in 39/40 (98%) 

patients. The most common adverse events (AEs) were fatigue, 

nausea, headache, rash, anemia, and asthenia. Two serious 

AEs were reported, neither of which was attributed to study 

drugs. One patient with a history of prior LT rejection discontinued 

treatment on day 86 due to low grade rejection revealed 

by liver biopsy, but achieved SVR12. Hemoglobin declines 


1085 

Utilizing claims data to understand the relationships 

between diagnosing, prescribing, and dispensing patterns 

for HCV patients in the pre and post sofosbivir 

eras 

Michel F. Denarie; Real world evidence Solutions, IMS Health, 

Yardley, PA 

UTILIZING CLAIMS DATA TO UNDERSTAND THE RELATION- 

SHIPS BETWEEN DIAGNOSING, PRESCRIBING, AND DIS- 

PENSING PATTERNS FOR HCV PATIENTS IN THE PRE AND 

POST SOFOSBUVIR ERAS BACKGROUND AND AIMS: The 

treatment of Hepatitis C (HCV) patients has undergone profound 

change since the introduction of sofosbuvir in late 2013. 

Many patients were warehoused until the launch of sofosbuvir 

because it was the first directly acting antiviral (DAA) to offer an 

interferon-free, short duration, highly tolerable and efficacious 

regimen. The study also determined the amount and duration 

of patient warehousing and the variations between ethnicities. 

The intent was to utilize claims data to demonstrate how HCV 

treatment dynamics have changed in the pre and post sofosbuvir 

era. DESIGN/METHODS: This 2 year retrospective study 

utilized US IMS claims data. Patients were selected for HCV 

diagnosis for a period of six months, then followed for a period 

of 12 months to determine the date of prescription, and then 

followed for an additional 6 months to ascertain whether they 

were actually dispensed the prescribed product(s). We limited 

the analysis to patients prescribed telaprevir, boceprevir, sofosbuvir 

and simeprevir; breaking into two patient cohorts: pre-sofosbuvir 

(Period 1/Jan 2012 - Nov 2013) and post-sofosbuvir 

(Period 2/Dec 2013 - Aug 2014) launch. Using these cohorts, 

we were able to determine an index diagnosis date, an index 

written date, and an index dispensed date. RESULTS: 11,256 

newly diagnosed HCV patients (40% females and 60% males) 

were included in the study. Results demonstrate the amount and 

length of patient warehousing increased dramatically in the 12 

months preceding the introduction of sofosbuvir and simeprevir 

(see Chart 1), most notably for patients prescribed sofosbuvir 

and simeprevir. During Period 1, 93% of simeprevir and 81% 

of sofosbuvir patients were dispensed their medication more 

than 12 months after the initial HCV diagnosis was recorded, 

while only 45% of simeprevir and 36% of sofosbuvir patients 

were dispensed their medication more than 12 months after 

initial diagnosis in Period 2. The time between the WRx (written 

scripts) to the DRx (dispensed script) did not materially change. 

Chart 1 CONCLUSIONS: This study displayed the drastic differences 

in patient warehousing between Periods 1 and 2, 

most notably for sofosbuvir and simeprevir treatments. Many 

newly diagnosed HCV patients waited for the new DAAs to be 

introduced to receive a prescription. Once prescribed, despite 

the high cost of treatment, the time between the WRx and the 

DRx did not materially increase, and remained mostly within 

one month indicating the benefits outweighed the cost. 

Disclosures: 

The following authors have nothing to disclose: Michel F. Denarie 

1086 

Long-Term Efficacy of Ombitasvir/Paritaprevir/r and 

Dasabuvir With or Without Ribavirin in HCV Genotype 

1-Infected Patients With or Without Cirrhosis 

Stefan Zeuzem 2 , Ira M. Jacobson 3 , Jordan J. Feld 4 , Heiner Wedemeyer 

5 , Xavier Forns 6 , Pietro Andreone 7 , Massimo Colombo 8 , 

David Bernstein 9 , Fred Poordad 10 , Christophe Hezode 11 , Thomas 

Podsadecki 1 , Wangang Xie 1 , Tami Pilot-Matias 1 , Regis A. 

Vilchez 1 , John M. Vierling 12 ; 1 AbbVie, North Chicago, IL; 2 J.W. 

Goethe University, Frankfurt, Germany; 3 Weill Cornell Medical 

College, New York, NY; 4 Toronto Centre for Liver Disease, University 

of Toronto, Toronto, ON, Canada; 5 Medizinische Hochschule 

Hannover, Hannover, Germany; 6 Liver Unit, Hospital Clinic, 

IDIBAPS and CIBEREHD, Barcelona, Spain; 7 Dipartimento di Scienze 

Mediche e Chirurgiche, University of Bologna, Bologna, Italy; 

8 Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, 

Università degli Studi di Milano, Milan, Italy; 9 North Shore University 

Hospital, Manhasset, NY; 10 The Texas Liver Institute/University 

of Texas Health Science Center, San Antonio, TX; 11 Henri Mondor 

University Hospital, AP-HP, Universite Paris-Est, Cr’eteil, France; 

12 Baylor College of Medicine, St. Luke’s Advanced Liver Therapies, 

Houston, TX 

Introduction: Treatment with the 3 direct-acting antiviral (3D) 

regimen of ombitasvir (OBV), paritaprevir (PTV, boosted with 

ritonavir [r]; identified by AbbVie and Enanta) and dasabuvir 

(DSV), with or without ribavirin (RBV), achieved sustained virologic 

response (SVR) rates between 95% and 100% across a 

broad range of hepatitis C virus (HCV) genotype (GT) 1-infected 

patients. In this analysis, we examined the efficacy through 

post-treatment week 48 of the 3D regimen in HCV GT1-infected 

patients with or without cirrhosis. Methods: Treatment-naïve and 

-experienced HCV GT1a- or GT1b-infected patients with and 

without compensated cirrhosis received the recommended 3D 

regimen of co-formulated OBV/PTV/r (25mg/150mg/100mg 

once daily) and DSV (250mg twice daily), ± weightbased 

RBV, for 12 or 24 weeks in six phase 3 studies. GT1b-infected 

patients without cirrhosis did not receive RBV as part of their 

treatment regimen. SVR at post-treatment week 12 (SVR12; 

HCV RNA







Gilead 

Pietro Andreone - Advisory Committees or Review Panels: Janssen-Cilag, Gilead, 

MSD/Schering-Plough, Abbvie; Speaking and Teaching: Gilead, BMS 












Gilead 







Gilead 

Thomas Podsadecki - Employment: AbbVie; Stock Shareholder: AbbVie 

Wangang Xie - Employment: AbbVie 


Regis A. Vilchez - Employment: AbbVie Inc. 









in lifetime risks of developing advanced liver disease (i.e. CC, 

DCC or HCC) and an 80% reduction in risks of LrD, regardless 

of treatment history or cirrhosis status. The lifetime risks of 

liver morbidity and mortality are also substantially lower in the 

overall coinfected patients treated with 3D±R than those treated 

with SOF+PR or SOF+R. In the naïve non-cirrhotic coinfected 

patients, treatment with 3D±R offers comparable reductions 

in lifetime risks of liver morbidity and mortality compared to 

SOF+LDV. Conclusion Compared with former or other current 

standards of care for treating GT1 HCV and HIV coinfected 

patients in the US, 3D±R offers favorable or comparable reductions 

in lifetime risks of liver morbidity and mortality. 

Lifetime risks of liver morbidity and mortality in GT1 HCV and HIV 

coinfected patients 

* Clinical trial data not available 

Disclosures: 





Suchin Virabhak - Consulting: AbbVie 

Scott J. Johnson - Consulting: AbbVie; Employment: Medicus Economics 

Hélène Parisé - Consulting: MedicusEconomics LLC, AbbVie ; Employment: Statlog 

Consulting Inc 



The following authors have nothing to disclose: Alice Wang 

1087 

Lifetime risks of liver morbidity and mortality in patients 

with chronic genotype (GT) 1 hepatitis C virus (HCV) 

and HIV coinfection treated with 3D±R (ombitasvir/ 

paritaprevir/ ritonavir, dasabuvir ± ribavirin) vs other 

standards of care in the US 

Sammy Saab 2 , Suchin Virabhak 3 , Scott J. Johnson 3 , Hélène 

Parisé 3 , Yuri Sanchez 1 , Alice Wang 1 , Timothy R. Juday 1 ; 1 HEOR, 

AbbVie, Mettawa, ID; 2 Pfleger Liver Institute, UCLA, Los Angeles, 

CA; 3 Medicus Economics Inc., Boston, MA 

Objective This study evaluated the lifetime risks of liver morbidity 

and mortality in patients with GT1 HCV and HIV coinfection 

treated with 3D±R for 12 or 24 weeks compared to other standards 

of care in the United States (US): sofosbuvir plus peg-interferon 

and ribavirin for 12 weeks (SOF+PR) and SOF+R for 

24 weeks in the overall coinfected population, and sofosbuvir 

plus ledipasvir for 12 weeks (SOF+LDV) in the treatment-naïve 

non-cirrhotic population. Methods A Markov model based on 

the natural history of liver disease progression estimated the 

risks of liver-related morbidity and mortality over a lifetime 

horizon for a cohort of patients with GT1 HCV and HIV coinfection. 

Baseline population characteristics and efficacy data 

were obtained from published clinical trials. Lifetime risks of 

compensated cirrhosis (CC), decompensated cirrhosis (DCC), 

hepatocellular carcinoma (HCC) and liver-related death (LrD) 

were analyzed. Treatment strategies included ‘no treatment’ 

(NT), as well as regimens that have been studied in Phase II or 

III clinical trials of coinfected patients, and either recommended 

by the latest guidelines or approved by the Food and Drug 

Administration. Results Compared to untreated GT1 HCV and 

HIV coinfected patients, 3D±R offers at least a 59% reduction 

1088 

Effect of Chronic Kidney Disease on the Pharmacokinetics 

of Ombitasvir, Paritaprevir, Ritonavir and Dasabuvir 

in Subjects with HCV Genotype 1 Infection 

Diana L. Shuster, Rajeev Menon, Daniel E. Cohen, Amit Khatri; 

AbbVie, North Chicago, IL 

Background and Objective: The all-oral interferon-free, 3 

direct acting antiviral (3-DAA) regimen of ombitasvir (OBV) 

(NS5A inhibitor) + paritaprevir (NS3/4A protease inhibitor 

identified by AbbVie and Enanta), coadministered with ritonavir, 

a pharmacokinetic enhancer (PTV/r), + dasabuvir (DSV) 

(NS5B non-nucleoside polymerase inhibitor) ± ribavirin (RBV) 

is being evaluated in HCV genotype (GT) 1-infected subjects 

with chronic kidney disease (CKD). No meaningful alterations 

in exposures were seen when the 3 DAAs were administered 

to HCV-uninfected subjects with renal impairment. The present 

analysis examines the effect of CKD Stage 4 and Stage 

5 on the pharmacokinetics of OBV, PTV/r and DSV in HCV 

GT1-infected subjects. Methods: Pharmacokinetic data from 

a Phase 2 study (N=38) in subjects with normal or mild renal 

impairment (subjects “without kidney disease”) were combined 

with preliminary data from a Phase 3b study in subjects with 

CKD Stage 4 (N=5) or Stage 5 on hemodialysis (N=14). In 

both studies subjects received OBV/PTV/r 25/150/100 mg 

QD and DSV 250 mg BID ± RBV for 12 weeks. Pharmacokinetic 

parameters and steady-state exposures of the 3-DAA 

regimen were estimated using population pharmacokinetic 

models. Results: CKD was not a significant covariate in the 

population pharmacokinetic analyses, and the safety profile 

of the 3 DAAs was similar in subjects with or without CKD. 

PTV and DSV exposures were comparable (


between subjects without kidney disease and CKD Stage 4. 

OBV and ritonavir exposures were ~80% and 200% higher, 

respectively, in CKD Stage 4 subjects in the limited number of 

subjects in this analyses. OBV and PTV exposures were comparable 

(7 log 10 

IU/mL. Reasons for ineligibility for ACTG 

A5327 were treatment initiated >24 weeks after HCV diagnosis 

(n=6); peak ALT


pre- or post-ACTG A5327 enrolment period (n=3) (1 man had 

2 reasons). All 12 men who were treated completed 12 weeks 

of SOF+RBV and 12 weeks of follow-up monitoring. Eleven 

(92%) achieved SVR 12. The one man with treatment failure, 

viral relapse between weeks 4 and 9 post-treatment, had a 

very low peak ALT (103 U/mL) and baseline VL higher than 

the median (6.12 log 10 

IU/mL). Irritability and insomnia were 

commonly reported but did not limit treatment. Conclusions 

SOF+RBV for 12 weeks was highly effective in the treatment of 

acute HCV in HIV-infected men in this “real-world” setting using 

broad enrollment criteria. Larger studies would be needed to 

confirm these findings, but with the subsequent availability of 

new oral agents, efforts should probably instead be directed 

toward developing shorter and RBV-free regimens. 

Disclosures: 




The following authors have nothing to disclose: Zachary Barbati, Andrew L. 

Foster, Tristan Morey, Samuel Turner 

1091 

Sofosbuvir in combination with simeprevir +/- ribavirin 

in genotype 4 hepatitis C patients with advanced fibrosis 

or cirrhosis: real-life experience from Belgium 

Christophe Moreno 1 , Luc Lasser 2 , Jean Delwaide 3 , Peter Starkel 4 , 

Wim Laleman 5 , Philippe Langlet 6 , Hendrik Reynaert 7 , Stefan 

Bourgeois 8 , Sergio Negrin Dastis 9 , Thierry Gustot 1 , Sven M. 

Francque 10 , Anja M. Geerts 11 , Christophe Van Steenkiste 12 , 

Chantal de Galocsy 13 , Collins Assene 13 , Hans Orlent 14 , Marcel 

Nkuize 15 , Delphine Degré 1 ; 1 Department of Gastroenterology, 

Hepatopancreatology and Digestive oncology, CUB Hôpital 

Erasme, Brussels, Belgium; 2 Department of Hepatogastroenterology, 

CHU Brugmann, Brussels, Belgium; 3 Department of Hepatogastroenterology, 

CHU Sart Tilman, Liège, Belgium; 4 Department 

of Gastroenterology, Hepatopancreatology and Digestive oncology, 

Cliniques Universitaires Saint-Luc, Brussels, Belgium; 5 Department 

of Liver and Biliopancreatic disorders, University Hospitals 

Leuven, Leuven, Belgium; 6 Department of Gastroenterology, Hepatopancreatology 

and Digestive oncology, CHIREC, Brussels, 

Belgium; 7 Department of Hepatogastroenterology, UZ Brussel, 

Brussels, Belgium; 8 Stuivenberg, ZNA, Antwerp, Belgium; 9 Grand 

hôpital de Charleroi, Charleroi, Belgium; 10 Department of Gastroenterology 

and Hepatology, University Hospital Antwerp, Edegem, 

Belgium; 11 Department of Hepatogastroenterology, Ghent University 

Hospital, Ghent, Belgium; 12 AZ Maria-Middelares, Ghent, 

Belgium; 13 Hôpitaux Iris Sud, Brussels, Belgium; 14 Department of 

Gastroenterology and Hepatology, AZ St Jan, Brugge, Belgium; 

15 Department of Hepatogastroenterology, CHU Saint-Pierre, Brussels, 

Belgium 

Background: All-oral, interferon-free regimens that combine 

direct-acting antiviral drugs have significantly advanced the 

treatment of hepatitis C (HCV), especially for genotype 1(G1) 

patients. However, efficacy and safety data of interferon-free 

regimens in HCV genotype 4 (G4) patients are scarce. In Belgium, 

Sofosbuvir (SOF) and Simeprevir (SMV) treatment is 

available since January 2015 for G4 patients with advanced 

fibrosis (F3-F4 METAVIR) for 12 weeks. Methods: analysis of 

HCV G4 patients receiving SOF and SMV treatment in Belgium. 

The aim of the study was to evaluate the safety and efficacy 

of the treatment. Results: 73 G4 patients were enrolled in 

this data collection including 32 (43.8%) patients with severe 

fibrosis F3 and 41(56.2%) cirrhotic patients. The study population 

comprised 58.9% male, 77.8% treatment experienced 

patients. Median age was 59 [51-66] years and 5 patients 

were HCV/HIV co-infected. 24 patients received the treatment 

associated with ribavirin, 11/32 (34.37%) of patients with 

advanced fibrosis and 13/41 (31.71%) of cirrhotic patients. In 

cirrhotic patients, median MELD and Child-Pugh score were 9 

[7-12.5] and 5 [5-6], 46.2% had platelet below 100.000/mm 

and 28.6% had albumin below 35 g/L. W4 HCV RNA was 

undetectable in 31.25% (15/48). 9 of the 15 patients with 

undetectable W4 HCV RNA received RBV. At W12, 100% 

(23/23) had HCV RNA below the limit of quantification, with 

6/23 still detectable. All SVR12 data will be available at the 

time of presentation. No patient experienced serious adverse 

event. Conclusions: these preliminary results in difficult-to-treat 

G4 HCV patients show that SOF/SIM +/- RBV treatment is safe 

and seems promising, in line with that was observed in G1 

HCV patients. 

Disclosures: 



Hendrik Reynaert - Advisory Committees or Review Panels: MSD, Gillead, Janssen, 

BMS, Abbvie, Norgine; Grant/Research Support: Roche 

Stefan Bourgeois - Advisory Committees or Review Panels: AbbVIe, Gilead; Consulting: 

Roche, MSD; Speaking and Teaching: Janssen, BMS 

The following authors have nothing to disclose: Luc Lasser, Jean Delwaide, Peter 

Starkel, Wim Laleman, Philippe Langlet, Sergio Negrin Dastis, Thierry Gustot, 

Sven M. Francque, Anja M. Geerts, Christophe Van Steenkiste, Chantal de 

Galocsy, Collins Assene, Hans Orlent, Marcel Nkuize, Delphine Degré 

1092 

Efficacy and safety of IFN-free, DAA-based treatment-regimens 

in patients with HCV-recurrence after 

Liver Transplantation: Real-life experience from Austria 

Sandra Beinhardt 1 , Ramona Al Zoairy 2 , Clarissa Freissmuth 1 , Karin 

Kozbial 1 , Stephanie Hametner 3 , Rafael Stern 1 , Rudolf E. Stauber 5 , 

Andreas Maieron 3 , Katharina Staufer 4 , Michael P. Strasser 6 , Heinz 

M. Zoller 2 , Ivo Graziadei 7 , Wolfgang Vogel 8 , Markus Peck-Radosavljevic 

1 , Michael Trauner 2 , Peter Ferenci 1 , Harald Hofer 1 ; 

1 Gastroenterology and Hepatology, Medical University of Vienna, 

Vienna, Austria; 2 Gastroenterology and Hepatology, Department 

of Internal Medicine II, Innsbruck, Austria; 3 Hosital Elisabethinen, 

Linz, Austria; 4 Department of Transplant Surgeries, Medical University 

of Vienna, Vienna, Austria; 5 Department of Gastroenerology 

and Hepatology, Medical University of Graz, Graz, Austria; 

6 Department of Gastroenerology and Hepatology, Paracelsus 

Medical University Salzburg, Salzburg, Austria; 7 Departmen of 

Internal Medicine, LKH Hall/Tirol, Hall/Tirol, Austria; 8 Department 

of Internal Medicine II, Division of Gastroenterology and Hepatology, 

Medical University of Innsbruck, Innsbruck, Austria 

Background&Aims: Patients after orthotopic liver transplantation 

(OLT) and hepatitis C virus-recurrence have high need 

of antiviral therapy (TX) due to accelerated progression of 

disease. IFN-free regimens revealed promising efficacy- and 

safety-data in clinical studies. This study aimed to evaluate 

real-life efficacy- and safety-data of DAA-regimens in patients 

with HCV-recurrence after OLT. Patients&Methods: 96 OLT-patients 

(10:liver-/kidney-transplantation; male:79/82.3%; 

age:59±14 years [mean±SD]; 34-78 [range]; GT-1:69 

[1a:17; 1b:49; 1g:1; undetermined:2]; GT3a:17; GT-4:8; 

missing:2) started all-oral DAA-regimens. Time from OLT to 

TX was 67±6 (mean±SD; range: -8–304) months. 8 patients 

had fibrosing cholestatic hepatitis (FHC), 49 cirrhosis (CPS- 

A:33; CPS-B/C:10; FIB4-Score/BL:6.9±8.7 [mean±SD]; TX-experienced:65/68%). 

Immunosuppressive-TX was in 37/39% 

patients based on calcineurin- and 55/57% on m-TOR-inhibitors. 

DAA-regimens were: Sofosbuvir(SOF)/ribavirin(RBV):26, 

SOF/Daclatasvir(DCV):43, SOF/Simeprevir(SIM):12, SOF/ 

Ledipasvir(LDV)±RBV:10; 3D-combination (Paritaprevir/r&Om-


bitasvir&Dasabuvir):4; SIM/DCV:1. HCV-RNA-quantification 

was performed with Abbott RealTime HCV ([ART], lower limit of 

quantification [LLOQ]:12IU/ml) or Roche COBAS AmpliPrep/ 

COBAS TaqMan assay (LLOQ:15IU/ml). Results: At present 

4/4.2% patients reached TX-week4, 14/15% end of treatment 

(EoT), 78/81% end of follow-up (12 weeks after TX). Overall 

SVR12-rate was 83% (65/78); table). 2 FCH-patients died 

at week 8 and 20, respectively; 2 interrupted treatment; one 

was transplanted at week10, the other hospitalized for hepatic 

encephalopathy; treatment was stopped at week8, the patient 

achieved SVR12. No episode of graft-rejection was observed. 

In SVR12-patients transaminases decreased compared to 

baseline (BL/SVR12:ALT:117±93 vs. 56±35; AST:113±95 

vs. 36±35,p


Christine Jacomet - Advisory Committees or Review Panels: ANSM; Consulting: 

CONVERGENCE; Grant/Research Support: JANSSEN, ROCHE, MSD; Speaking 

and Teaching: ABBOTT, BMS, MSD, JANSSEN, VIIV, GILEAD 

Claudine C. Duvivier - Grant/Research Support: Gilead Sciences 

Laurent Cotte - Grant/Research Support: MSD, ViiV 

The following authors have nothing to disclose: Isabelle Poizot-Martin, Alissa 

Naqvi, Véronique Obry-Roguet, Eric Billaud, Antoine Chéret, David Rey, Pascal 

Pugliese, Pierre Pradat 

1094 

Sofosbuvir Plus Ribavirin Without Interferon For Treatment 

of Acute Hepatitis C Virus Infection in HIV-1 

infected Individuals (SWIFT-C) 

Susanna Naggie 1 , Kristen M. Marks 2 , Michael Hughes 3 , Daniel S. 

Fierer 4 , Arthur Y. Kim 5 , Kimberly Hollabaugh 3 , Jennifer Kiser 6 , Jhoanna 

Roa 3 , Bill Symonds 7 , Diana M. Brainard 8 , John G. McHutchison 

8 , Marion G. Peters 9 , Raymond T. Chung 5 ; 1 Duke University 

Medical Center, Durham, NC; 2 Weill Cornell, New York, NY; 

3 Harvard, Cambridge, MA; 4 Mount Sinai School of Medicine, 

New York, NY; 5 Massachusetts General Hospital, Boston, MA; 

6 University of Colorado, Denver, CO; 7 Roivant, New York, NY; 

8 Gilead Sciences, Inc, Foster City, CA; 9 University of California, 

San Francisco, San Francisco, CA 

Background: The role of interferon-free, oral direct acting antivirals 

(DAA) for the treatment of acute hepatitis C virus (HCV) 

infection in HIV-infected persons has not been explored. Methods: 

SWIFT-C is a multicenter, single arm trial of the AIDS Clinical 

Trials Group (ACTG) investigating the safety and efficacy 

of 12 weeks of sofosbuvir (400mg/day) and weight based 

ribavirin (1000mg


dL (IQR: 0.5 – 0.9 mg/dL). Of the patients with data available 

12-weeks after the end-of-treatment, 43/48 (90%) achieved 

SVR12. Of the 5 patients who failed, 4 relapsed within the first 

4 weeks after the end-of-treatment. One patient was sent to hospice 

after a hospitalization for acute cholangitis with elevated 

total bilirubin, rectal bleeding, and worsening renal function. 

The average cost of pharmaceuticals per patient was $90,234. 

The average cost-per-SVR was $100,727. Compared to treatment 

with SMV/SOF, the cost-per-SVR with LDV/SOF was on 

average $68,337 lower. Conclusions: We observed high SVR 

rates (90%) with LDV/SOF in standard clinical practice. The 

cost-per-SVR was $100,000 and was approximately $68,000 

less than previous treatments. Future investigations of real-world 

SVR rates are needed. 

Disclosures: 

Kian Bichoupan - Consulting: Janssen, Gilead 

Alyson Harty - Advisory Committees or Review Panels: Gilead; Consulting: Gilead, 

Jannsen, Acaria Pharmacy, Abbvie 

David B. Motamed - Advisory Committees or Review Panels: Gilead Pharmaceuticals 

Viktoriya Khaitova - Advisory Committees or Review Panels: Gilead, Johnson 

and Johnson 

Charissa Y. Chang - Consulting: Gilead, Vertex, Onyx 

Jennifer Leong - Advisory Committees or Review Panels: Gilead; Consulting: 

Bayer 











Albert Min - Consulting: Bristol Myers Squibb, Gilead, Janssen, Merck; Grant/ 

Research Support: Bristol Myers Squibb, Gilead; Speaking and Teaching: Bristol 

Myers Squibb, Gilead 




The following authors have nothing to disclose: Daniel J. Waintraub, Neal M. 

Patel, Sweta Chekuri, Joshua Hartman, Alicia Stivala, Angela Woody, Meena 

B. Bansal, Priya Grewal, Ritu Agarwal, Gene Y. Im, Lawrence Liu, Joseph A. 

Odin, Nancy Bach, Thomas D. Schiano, Ponni V. Perumalswami, Lan S. Wang, 

Jahnavi Naik 

1096 

IFN-λ inhibits miR-122 transcription through a Stat3- 

HNF4a inflammatory feedback loop in an IFN-α resistant 

HCV cell culture system 

Fatma Aboulnasr 1 , Sidhartha Hazari 1 , Partha K. Chandra 1 , Pauline 

Ferraris 1 , Rajesh Panigrahi 1 , Srinivas Chava 1 , Ramazan Kurt 1 , 

Kyoungsub Song 1 , Asha Dash 1 , Luis A. Balart 2 , Tong Wu 1 , Srikanta 

Dash 1,2 ; 1 Pathology, Tulane University, New Orleans, LA; 

2 Medicine, Division of Gasteroenterology and Hepatology, Tulane 

University, New Orleans, LA 

Background: HCV replication in persistently infected cell culture 

remains resistant to IFN-α /RBV combination treatment, 

whereas IFN-λ1 induces viral clearance. The antiviral mechanisms 

by which IFN-λ1 induces sustained HCV clearance 

have not been determined. Aim: To investigate the mechanisms 

by which IFN-λ clears HCV replication in an HCV cell 

culture model. Methods: IFN-α sensitive (S3-GFP) and resistant 

(R4-GFP) cells were treated with equivalent concentrations of 

either IFN-λ1 or IFN-α.The relative antiviral effects of IFN-α 

and IFN-λ1 were compared by measuring the HCV replication, 

quantification of HCV-GFP expression by flow cytometry, and 

viral RNA levels by real time RT-PCR. Activation of Jak-Stat 

signaling, interferon stimulated gene (ISG) expression, and 

miRNA-122 transcription in S3-GFP and R4-GFP cells were 

examined. Results: We have shown that IFN-λ1 induces HCV 

clearance in IFN-α resistant and sensitive replicon cell lines 

in a dose dependent manner through Jak-Stat signaling, and 

induces STAT1 and STAT2 activation, ISRE-luciferase promoter 

activation and ISG expression. Stat3 activation is also involved 

in IFN-λ1 induced antiviral activity in HCV cell culture. IFNλ1 

induced Stat3 phosphorylation reduces the expression of 

hepatocyte nuclear factor 4 alpha (HNF4α) through miR-24 in 

R4-GFP cells. Reduced expression of HNF4α is associated with 

decreased expression of miR-122 resulting an anti-HCV effect. 

Northern blot analysis confirms that IFN-λ1 reduces miR-122 

levels in R4-GFP cells. Our results indicate that IFN-λ1 activates 

the Stat3-HNF4a feedback inflammatory loop to inhibit miR- 

122 transcription in HCV cell culture. 

Disclosures: 

Luis A. Balart - Advisory Committees or Review Panels: abbvie, Genentech; 

Grant/Research Support: Merck, Genentech, Bayer, conatus, Ocera, Hyperion, 

Gilead Sciences, Bristol Myers Squibb, abbvie, Vertex, Merck, Genentech, 

Bayer, Conatus, Ocera, Hyperion, Gilead Sciences, Bristol Myers Squibb, 

Mochida, Eisai, Vertex, takeda, GI Dynamics, tobira; Speaking and Teaching: 

Merck, Merck, Merck, Merck, Abbvie, janssen 

The following authors have nothing to disclose: Fatma Aboulnasr, Sidhartha 

Hazari, Partha K. Chandra, Pauline Ferraris, Rajesh Panigrahi, Srinivas Chava, 

Ramazan Kurt, Kyoungsub Song, Asha Dash, Tong Wu, Srikanta Dash 

1097 

Efficacy of Daclatasvir/Asunaprevir According to Resistance 

Associated Variants in Chronic Hepatitis C Genotype1b 

Etsuko Iio 1 , Noritomo Shimada 2 , Hiroshi Abe 3 , Masanori Atsukawa 

4 , Kai Yoshiza 5 , Koichi Takaguchi 6 , Yuichiro Eguchi 7 , Hideyuki 

Nomura 8 , Tomoyuki Kuramitsu 9 , Jong-Hon Kang 10 , Takeshi 

Matsui 10 , Noboru Hirashima 11 , Atsunori Kusakabe 12 , Yasuhito 

Tanaka 1 ; 1 Nagoya City University Graduate School of Medical 

Sciences, Nagoya, Japan; 2 Ootakanomori Hospital, Kashiwa, 

Japan; 3 Jikei University School of Medicine Katsushika Medical 

Center, Tokyo, Japan; 4 Nippon Medical School Chiba Hokusoh 

Hospital, Chiba, Japan; 5 Machida Municipal Hospital, Tokyo, 

Japan; 6 Kagawa Prefectural Central Hospital, Takamatsu, Japan; 

7 Saga University Hospital, Saga, Japan; 8 Shin-Kokura Hospital, 

Kitakyushu, Japan; 9 Kuramitsu Clinic, Akita, Japan; 10 Teine Keijinkai 

Hospital, Sapporo, Japan; 11 National Hospital Organization 

Nagoya Medical Center, Nagoya, Japan; 12 Nagoya Red 

Cross Hospital, Nagoya, Japan 

[Background/Aims] Interferon (IFN)-free daclatasvir (DCV) 

and asunaprevir (ASV) therapy for 24 weeks improved efficacy 

and safety outcomes for patients with hepatitis C virus 

(HCV) infection. The aim of the present study is to assess the 

treatment outcome according to resistance associated variants 

(RAVs) in NS3/NS5A regions among patients with chronic 

HCV genotype 1b infection (HCV-1b). [Methods] 612 patients 

with HCV-1b were enrolled at multi centers in Japan. This 

study protocol was approved by the appropriate institutional 

ethics review committees and written informed consents were 

observed. Direct sequencing in NS5A region was performed 

for all 612 patients before DCV/ASV therapy. 393 chronic 

hepatitis (CH) and 219 liver cirrhosis (LC) patients. The median 

age was 71 (30-87) and male was 267 (43.6%). The median 

platelet count was 12.6×10 4 /μL and HCV-RNA was 6.1 log 

IU/mL. 36 patients had experience of Peg-IFN/RBV/Protease 

inhibitors (PI) combination therapy, and 27 with simeprevir 

(SMV). [Results] Direct sequencing analysis of 612 patients 

administered DCV/ASV therapy showed the existence of 31M 

(2.4%), 93H (4.1%), and 31M/93H (0.3%) variants in NS5A 

region. The 531 patients received over the 4 weeks of DCV/


ASV; 75.0% (398/531) achieved rapid virologic response 

(RVR) and 97.0% (450/482) became undetectable until 8 

week. There was no significant differences of RVR among 

IL28B genotypes. 76.9% (380/494) with wild-type NS5A-Y93 

achieved RVR, whereas only 54.2% (13/24) with Y93H substitution 

achieved RVR. In 93Y group, there were 22 viral breakthrough 

and 3 relapsers, Among these patients with treatment 

failure, 40.0% (10/25) had SMV treatment history. On the 

other hand, among patients with Y93H at baseline, 5 patients 

(20.1%) had virologic failure and 1 patient experienced 

relapse, resulting in emergence of NS5A-Y93 and NS3-D168 

variants. Finally, SVR (sustained virologic response) 4 was 

89.6% in Y93, whereas 45.5% in Y93H, suggesting that RVR 

and SVR were higher in 93Y than 93H group. Interestingly, 

multiple RAVs such as L31M/Q54H/Y93H in NS5A were 

emerged by treatment failure. The number of RAVs were 0/ 1/ 

2/ 3=57/ 36/ 7/ 0 (%) at baseline, and 0/ 14/ 57/ 29 (%) 

at the time of virologic failure; patients with multiple RAVs in 

NS5A region increased from 7% to 86%. [Conclusion] History 

of SMV therapy and pre-existing NS5A-Y93H substitution were 

associated with breakthrough during DCV/ASV therapy, resulting 

in emergence of multiple RAVs. As the multi-RAVs influence 

SVR rate by DAAs therapy including NS5A inhibitors, patients 

with RAVs at baseline will be required assessment for optimizing 

future DAAs therapies. 

Disclosures: 



Squibb 

The following authors have nothing to disclose: Etsuko Iio, Noritomo Shimada, 

Hiroshi Abe, Masanori Atsukawa, Kai Yoshiza, Koichi Takaguchi, Yuichiro Eguchi, 

Hideyuki Nomura, Tomoyuki Kuramitsu, Jong-Hon Kang, Takeshi Matsui, 

Noboru Hirashima, Atsunori Kusakabe 

1098 

Ribavirin levels at treatment weeks (TW) 4-6 can predict 

sustained viriological response (SVR12) in HCV cirrhotic 

patients treated with all oral direct–acting antiviral therapy 

(DAAs). 

Suman Verma 1 , Ivana Carey 1 , Kate E. Childs 1 , Andrew Ayers 1 , 

Phillip Morgan 1 , Aisling B. Considine 1 , Kath Oakes 1 , Michael 

A. Heneghan 1 , Graham R. Foster 2 , Kosh Agarwal 1 ; 1 Institute of 

liver studies, Kings College Hospital, London, United Kingdom; 

2 The Blizard Institute, Queen Marys University of London, London, 


Background Ribavirin remains an important adjunct to DAAs 

in end-stage cirrhotic patients (Child–Pugh(CP) B7 and above). 

However ribavirin associated side-effects can be challenging 

to manage in this population (CUPIC). Hence this study aimed 

to investigate whether plasma ribavirin level monitoring in 

this advanced liver disease population can optimise SVR12. 

Method 46 patients, with CP B7-C10 cirrhosis, receiving ribavirin, 

Sofosbuvir and either Ledipasvir or Daclatasvir had 

serial trough plasma ribavirin measurements at TW2, 4, 6 and 

8 using a validated liquid chromatography assay. At TW0, 

a median of 1000mg/day (range 1000mg/day-1200mg/ 

day) ribavirin was administered to all patients. Despite high 

inter-patient variability in ribavirin levels with the same dose, 

there was low intra-patient variation with steady-state achieved 

between TW4-TW6 and the median dose remained 1000mg/ 

day. Hence levels at this timepoint were compared between 

those acheiving SVR12 vs. relapsing post treatment. All statistical 

analysis was performed using SPSS. Results 37 patients 

had SVR12 and 9 relapsed, but no significant difference in CP, 

UKELD, MELD, prescribed ribavirin dose (median 1000mg/ 

day) or ribavirin dose/kg at TW0 and TW4-6 was identified 

(Table 1). 25 patients with SVR12 and 2 relapsers had ribavirin 

dose reduction during 12 weeks of therapy but none 

required erythropoetin or blood transfusion support. Age, CP, 

UKELD, MELD, and genotype showed no significant correlation 

with SVR12 or relapse. However, the median plamsa ribavirin 

level was higher in the SVR12 cohort vs. relapsers (2.4mg/L 

and 1.4mg/L respectively (p=0.003) and levels ≥1.7mg/L at 

TW4-TW6 correlated with SVR12 achievement (PPV 89.19%; 

NPV 77.18%)(p=0.001 Fisher’s exact test). Ribavirin dose 

reduction thereafter did not influence SVR12 development. 

Conclusion Ribavirin levels at 4-6 weeks of a 12 week DAA 

regimen are predictive of SVR12 in this advanced liver disease 

cohort. Ribavirin plasma measurements can be a useful adjunct 

in delivering optimal SVR12 and tolerability in this advanced 

challenging population. 

SVR12 and relapser cohort demographics (median and ranges 

shown) 

Disclosures: 

Ivana Carey - Grant/Research Support: Gilead, Roche; Speaking and Teaching: 

BMS 







Kosh Agarwal - Advisory Committees or Review Panels: Gilead, Novartis, 

Abbott; Grant/Research Support: Roche, MSD; Speaking and Teaching: BMS, 

Astellas, Janssen 

The following authors have nothing to disclose: Suman Verma, Kate E. Childs, 

Andrew Ayers, Phillip Morgan, Aisling B. Considine, Kath Oakes, Michael A. 

Heneghan 

1099 

Frequency of Renal Impairment in Patients With Hepatitis 

C Infection Treated With Sofosbuvir-based Antiviral 

Regimens 

Saeed Almarzooqi 1 , Jagpal S. Klair 2 , Joel G. Karkada 1 , Raoel 

Maan 1 , Orlando Cerocchi 1 , Matthew Kowgier 1 , Sherrie M. Harrell 

2 , Kimberly Rhodes 2 , Harry L. Janssen 1 , Jordan J. Feld 1 , Andres 

Duarte-Rojo 2 ; 1 Toronto Center for Liver Disease, Toronto, ON, Canada; 

2 University of Arkansas for Medical Sciences, Little Rock, AR 

BACKGROUND: Renal impairment (RI) was a relevant adverse 

event noted in 5-7% of hepatitis C patients treated with the 

protease inhibitors boceprevir/telaprevir (BOC/TPV), although 

it was infrequent with peginterferon/ribavirin (PR). RI has been 

very rarely reported in trials with newer direct-acting antivirals 

such as simeprevir (SMV) and sofosbuvir (SOF). However, SOF 

is renally cleared and it might cause RI in patients with pre-existing 

kidney disease. We aimed to examine the rate of RI in 

patients treated with SOF-based therapies, and to compare it 

to that of BOC/TPV. METHODS: Patients treated with any SOFbased 

or BOC/TPV-based therapies at two university-based 

referral centers were included. Demography, presence of cirrhosis, 

ascites, original MELD score, SVR12, comorbidities, 

use of nephrotoxic drugs and on-treatment changes in creatinine 

(Cr) were obtained from medical records. Posttransplant


patients were excluded. RI was defined as an increase in Cr 

≥50% from baseline while on-treatment. Mann-Whitney U, 

Kruskal-Wallis, chi-square, and logistic regression were used 

for analysis. RESULTS: A total of 456 patients (55±9 years, 

61% males, cirrhotic 50%, HIV 3%) were included (G1: 78%, 

G2: 13%, G3: 7%, G4&6: 2%). BOC/TPV was used in 224 

patients (50%); SOF+PR in 76 (17%); SOF+R in 75 (17%); 

SMV/SOF±RBV in 66 (15%); and LDV/SOF in 11 (2%). 

Patients on PR-based therapies were less likely cirrhotic (44% vs 

60%, p=0.001) and had lower MELD scores when compared 

to those on all-oral therapies (4 [2-6] vs 6 [3-9], p=0.0001). In 

total, 95% had a baseline eGFR ≥60 mL/min (Cr 0.83±0.16 

mg/dL). Among them, RI was noted in 7% of patients (n=15) 

on BOC/TPV, 5% of SOF-PR (n=3), and 4% of all-oral regimens 

(n=5), (p=0.4). On-treatment CrMax in patients on BOC/ 

TPV was 1.4 (1.2-2) mg/dL, on SOF-PR 2 (1.2-2), and on alloral 

regimens 2 (1.35-3.2) (p=0.04). Although cirrhosis and 

MELD were linked to RI on univariate analysis, only ascites 

(OR=3.16 [1.14-8.92]) and preexisting proteinuria (OR=5.74 

[2.04-16.15]) remained significant in multivariate models. In 

all cases, serum Cr returned to baseline after stopping therapy. 

SVR12 did not differ between those who did or did not 

develop RI (88% vs 86%, p=0.9). CONCLUSIONS: Reversible 

RI in SOF-based therapies was seen in 4-5% in this cohort with 

a high prevalence of cirrhosis, similar to rates reported with 

BOC/TPV-based regimens. RI was more frequent in patients 

with advanced liver disease, mainly in the presence of ascites 

and in those with preexisting kidney injury. Monitoring of 

renal function and standard nephroprotective measures are 

suggested when considering SOF-based regimens. 

Disclosures: 












The following authors have nothing to disclose: Saeed Almarzooqi, Jagpal S. 

Klair, Joel G. Karkada, Orlando Cerocchi, Matthew Kowgier, Sherrie M. Harrell, 

Kimberly Rhodes 

1100 

Prospective Study for The Efficacy of Sofosbuvir and 

Simeprevir ± Ribavirin in Hepatitis C Genotype 1 and 

4 Compensated Cirrhotic Patients. Single Center Study 

and Real Life Experience 

Zeid Kayali 1,2 , Cory Amador 2 , Andrew Lowe 2 , Besher Ashouri 1,2 , 

Katherine Lam 2 , Warren N. Schmidt 3 ; 1 Inland Empire Liver Foundation, 

Rialto, CA; 2 Arrowhead Regional Medical Center, Colton, 

CA; 3 University of Iowa Hospital and Clinics, Iowa City, IA 

Sofosbuvir (SOF) and simeprevir (SMV) ± ribavirin (RBV) 

have been used widely to treat hepatitis C genotype 1 and 4 

patients and showed 94% SVR12 in patients with stage 3 and 

4 fibrosis (COSMO study). The efficacy of this regimen, however, 

has never been validated in a large number of cirrhotic 

patients from a community, multiracial setting. The goal of this 

study was to investigate the performance of this regimen in a 

large, diverse group of difficult to treat cirrhotic patients. Methods: 

This was a prospective, open-label single center study. 

Enrolled subjects were consecutive patients with cirrhosis, genotype 

1 and 4, including both naïve and treatment experienced 

patients. Cirrhosis (Stage 4 fibrosis) was diagnosed based on 

biopsy and/or Fibrosure test. Decompensated cirrhotics, HIV 

co-infected, hepatocellular carcinoma, and active substance 

abuse patients were excluded. Duration of treatment was either 

24 weeks without RBV or 12 weeks with ribavirin. Primary 

end point was SVR12. Secondary end points were safety and 

relapse. Results: One hundred eight patients were enrolled. 

Mean age 54 year, 63 (58%) were male, 36 (33%) Caucasians, 

25 (21%) Black and 50 (46%) Hispanics. mean BMI 33, 

86 (79.6%) patients had HCV GT1a, and 7 (7%) had GT4, 

mean baseline viral load was 2.2 x 10 6 IU/ml, 59 (54%) were 

treatment naive. All patients completed treatment. No Grade 4 

adverse events were reported. HCV RNA was undetected in 90 

patients (83%) at end of treatment. Eighty-four patients (77%) 

achieved SVR12 and 24 patients (23%) relapsed. There was 

no change in MELD score or worsening decompensation at end 

of treatment. Univariate regression analysis showed that BMI 

(33) and Black race were independent factors associated with 

relapse (p=0.004,95%CI 1-1.22) and (p=0.025,95%CI 0.03- 

0.23) respectively. Duration of treatment, sex, age, baseline 

viral load, Geno1 subtypes and MELD score were not independent 

factors that predict relapse. Conclusion: In this large 

study of difficult to treat compensated cirrhotics patients, SOF 

and SMV ± RBV regimen was well tolerated but had lower SVR 

12 than previously reported. Surprisingly Black race and BMI 

were independent factors for relapse and these variables need 

to be considered when deciding the best regimen for difficult 

to treat patients. 

Disclosures: 

Zeid Kayali - Consulting: Abbvie; Grant/Research Support: Merck, Gilead 

Warren N. Schmidt - Consulting: gilead 

The following authors have nothing to disclose: Cory Amador, Andrew Lowe, 

Besher Ashouri, Katherine Lam 

1101 

Sofosbuvir + Ledispasvir Combination Therapy for 

Recurrent Hepatitis C in Liver Transplant Recipients: A 

Real-Life Multicenter Experience 

Ryan M. Kwok 1 , Suzanne Robertazzi 1 , Helen S. Te 2 , Joshua Wiegel 

3 , Joseph Ahn 3 , Janet Gripshover 4 , Darryn R. Potosky 4 , Amber 

Tierney 5 , Mohamed A. Hassan 5 , Rohit Satoskar 1 , Coleman I. 

Smith 1 ; 1 Transplant Institute, Medstar Georgetown University Hospital, 

Washington, DC; 2 Medicine, University of Chicago, Chicago, 

IL; 3 Oregon Health and Science University, Portland, OR; 

4 Division of Gastroenterology and Hepatology, University of Maryland 

School of Medicine, Baltimore, MD; 5 Gastroenterology Division, 

University of Minnesota Medical School, Minneapolis, MN 

Background Recurrent hepatitis C (HCV) after liver transplant 

(LT) is associated with significant morbidity and mortality. 

Second generation direct acting antiviral medications such as 

sofosbuvir / ledipasvir (SOF/LDV) have a high rate of sustained 

virologic response in treating non-cirrhotic and pretransplant 

HCV. Data on the efficacy and safety of SOF/LDV in the post-LT 

patient are limited. Aim: To evaluate the safety and efficacy of 

the SOF/LDV therapy in patients with HCV recurrence after LT. 

Methods All post-LT patients with recurrent HCV . previously or 

currently being treated with SOF/LDV with or without ribavirin 

for 12-24 weeks were identified at five centers. Sustained 

virologic response at 12 weeks (SVR12) after treatment was 

determined. On-treatment response, graft function, changes in 

immunosuppression, adverse events (AE), and survival were 

recorded. Results 128 patients were included: 76% male, 

65% Caucasian with a mean age of 60.9 ±6.8 years. 70% of 

patients were genotype (GT) 1a, 23% GT 1b, 1 patient was 

a GT 2, 2 patients were GT 4, 1 patient had both GT 1a/4. 

10% had kidney transplant, 52% had failed prior therapy, and


18% had F3-4 fibrosis. The median time from LT to SOF/LDV 

was 1398 days (15-7593 days). Tacrolimus was the primary 

immunosuppression in 86% of patients. 59% (44/74) of treatment 

week 4 (TW4) and 100% (51/51) of TW12 patients had 

undetectable viral load. 100% (12/12) achieved SVR12. 15% 

required changes in immunosuppression during or after treatment 

with 1 patient experiencing mild rejection at TW4 that 

was controlled with an increase in oral immunosuppression. 

Adverse events were reported in 15% of patients with constitutional 

symptoms being the most common (9%) and cough, 

rash, and headaches in 2%. In the 44 patients who received 

ribavirin, 50% developed anemia with only 3 patients requiring 

blood transfusion or erythropoeisis-stimulating agents. No 

serious adverse events were reported. Conclusions SOF / LDV 

is effective and safe in post-LT patients with recurrent HCV infection 

across a spectrum of genotypes. No significant changes 

in immunosuppression or graft function were noted. Adverse 

events were mild and were predominantly associated with the 

use of ribavirin. Additional SVR data will be reported. Further 

large, prospective trials are required to confirm these results. 

Disclosures: 



Joseph Ahn - Advisory Committees or Review Panels: gilead; Grant/Research 

Support: bms 

Janet Gripshover - Advisory Committees or Review Panels: Gilead, Abbvie; Consulting: 

Abbvie; Speaking and Teaching: Gilead, Abbvie 

Darryn R. Potosky - Advisory Committees or Review Panels: Gilead, Abbvie 

Mohamed A. Hassan - Speaking and Teaching: GILEAD 

Coleman I. Smith - Advisory Committees or Review Panels: Vertex, Gilead, Janssen; 

Grant/Research Support: Gilead, Abbvie, Janssen, Salix, BMS, Merck, Intercept 

Pharma, Lumena Pharma; Speaking and Teaching: Merck, Vetex, Gilead, 

Bayer/Onyx, BMS, Abbvie, Janssen 

The following authors have nothing to disclose: Ryan M. Kwok, Suzanne Robertazzi, 

Joshua Wiegel, Amber Tierney, Rohit Satoskar 

1102 

Hepatitis C treatment with Sofosbuvir and NS5A inhibitor 

in patients with an addiction 

Jean-Baptiste Trabut 5 , Camille Barrault 1 , Hélène Charlot 2 , Damien 

Carmona 3 , Willy Kini-Matondo 2 , Franck Questel 4 , Christophe 

Hezode 6 ; 1 Addictologie, CHIC, Créteil, France; 2 Pharmacie, 

Hôpitaux Universitaires Henri Mondor, Créteil, France; 3 Centre 

Epice, Créteil, France; 4 Addictologie, Hôpital Fernand Widal, 

Paris, France; 5 Addictologie, Hôpitaux Universitaires Henri Mondor, 

Créteil, France; 6 Hépatologie, Hôpitaux Universitaires Henri 

Mondor, Créteil, France 

Background: Hepatitis C virus (HCV) infection is highly prevalent 

in patients with an addiction. Access to HCV treatment 

is reduced in those patients due to the poor tolerance of interferon-based 

therapies. There are very few data on the use of 

“interferon-free” oral combinations in that population. Methods: 

patients infected with HCV and advanced fibrosis followed in 

four addictions care centres were considered for a treatment 

combining sofosbuvir and an NS5A inhibitor (daclatasvir or 

ledipasvir) during 12 weeks (24 weeks for cirrhotic patients 

with genotype 3). A multidisciplinary team including addiction 

specialists, hepatologists and pharmacists reviewed all indications 

and decided the practical implementation of the treatments. 

Results: Treatment has been introduced in 39 patients 

(Male: 87%, Median age: 51, cirrhotic: 71%). Two third of 

the patients received a substitution for opioid dependence and 

78% had an alcohol use disorder. Most patients (62%) suffered 

from some forms of psychosocial vulnerability. Treatment has 

been completed to date in 22 patients with an end of treatment 

viral clearance in all cases. No virological relapse has been 

reported so far and sustained virological response has been 

established in 15 patients. Updated results will be presented 

at the congress. Conclusion: Our preliminary results suggest 

that the use of new oral combinations for the treatment of HCV 

infection could be highly effective in the addiction care setting. 

Disclosures: 


Gilead 

The following authors have nothing to disclose: Jean-Baptiste Trabut, Camille 

Barrault, Hélène Charlot, Damien Carmona, Willy Kini-Matondo, Franck Questel 

1103 

Determination of on-treatment pharmacokinetics of 

sofosbuvir and ledipasvir in patients with decompensated 

cirrhosis - the need for real world PK studies 

Omar El-Sherif 1,6 , Diarmaid D. Houlihan 2 , Stephen Stewart 3 , 

Colm J. Bergin 4,6 , Liam J. Fanning 8 , Susan McKiernan 1,6 , Orla 

M. Crosbie 7 , Suzanne Norris 1,6 , Saye H. Khoo 5 ; 1 Department of 

Hepatology, St. James’s Hospital, Dublin, Ireland; 2 Liver Unit, St 

Vincents Hospital, Dublin, Ireland; 3 Mater Hospital, Dublin, Ireland; 

4 Department of Infectious Diseases, St James’s Hospital, Dublin, 

Ireland; 5 University of Liverpool, Liverpool, United Kingdom; 

6 Trinity College, Dublin, Ireland; 7 Cork University Hospital, Cork, 

Ireland; 8 University College Cork, Cork, Ireland 

Introduction: With the availability of interferon-free direct acting 

antiviral therapy (DAA) for hepatitis C, patients with decompensated 

cirrhosis can now be considered for antiviral therapy. 

The combination of sofosbuvir-ledipasvir with ribavirin was 

associated with high sustained virological response rates (SVR) 

in decompensated cirrhotics in the SOLAR-1 trial, although real 

world outcomes are awaited. It is unclear whether interindividual 

pharmacokinetic variability affects treatment outcome. 

Methods: Plasma trough samples were prospectively collected 

in 35 decompensated cirrhotic patients receiving 12 weeks 

of sofosbuvir-ledipasvir and ribavirin therapy for HCV genotype 

1 infection. GS-331007 (major circulating metabolite of 

sofosbuvir) and ledipasvir plasma trough samples were collected 

at treatment days 7, 14, 28, 84 and measured using 

validated LC-MS/MS. Mean age was 53.6 + 10.3 and 62% 

were male. The median MELD score was 10 + 3.3. Trough 

GS-331007 and ledipasvir concentrations were compared in 

patients achieving a rapid virologic response (RVR) vs no RVR 

and SVR4 vs non-SVR4. Results: All 35 patients achieved an 

on-treatment virologic response, and the SVR4 rate per protocol 

is 82%. GS-331007 C min 

ranged from 47 – 1097 ng/ml, 

and ledipasvir C min 

ranged from 36.1 – 366ng/ml. There was 

no relationship between trough GS-331007 levels and RVR 

or SVR. A trend towards higher mean day 28 ledipasvir C min 

levels in patients achieving SVR4 compared to early relapsers 

(182ng/ml vs 102.47ng/ml – p=0.062) was observed. Conclusion: 

The relationship between ledipasvir C min 

and treatment 

response needs to be examined in larger number of patients. 

These data highlight the potential utility of “real world” pharmacokinetic 

studies in optimising therapy and predicting treatment 

reponse in patients with advanced liver disease. 

Disclosures: 

Colm J. Bergin - Advisory Committees or Review Panels: Janssen, MSD, BMS, 

Pfizer; Grant/Research Support: MSD, Janssen, GSK, Abbott 

Saye H. Khoo - Grant/Research Support: Merck, Janssen, Gilead, ViiV 

The following authors have nothing to disclose: Omar El-Sherif, Diarmaid D. 

Houlihan, Stephen Stewart, Liam J. Fanning, Susan McKiernan, Orla M. Crosbie, 

Suzanne Norris


1104 

Real Life Experience of Direct Acting Anti-Viral Therapy 

for Hepatitis C infection in North East of Scotland 

Pauline Dundas, Shirley English, Lorna Bailey, Ashis Mukhopadhya, 

Balasubramaniam Vijayan, Andrew Fraser, Lindsay 

McLeman; Gastroenterology, Aberdeen Royal Infirmary, Aberdeen, 


There have been several clinical trials reported for new direct 

acting antiviral (DAA) drugs for hepatitis C (HCV) infection, 

but very few reports on ‘real life data’. We report our single 

centre experience of 140 patients initiated on the current generation 

of DAA agents in the North East of Scotland between 

October 2014 and May 2015. The choice and duration of 

regimen was decided according to local guidelines taking into 

consideration genotype, stage of disease, viral load, HIV coinfection, 

drug interaction, previous treatment and drug licences. 

Regimens included Simeprevir, Sofosbuvir, Daclatasvir and 

Sofosbuvir/Ledipasvir combinations. Sustained viral response 

(SVR) 4 week data will be available by September 2015 and 

12 week by November 2015. The regimens and patient characteristics 

are described in table 1. We report on 142 patient 

episodes of which 116 (82%) were initiated on an IFN free 

regimen. The mean age was 47 years (SD 10.9, range 20-79) 

and 109 (77%) were male. Eight (6%) patients were co-infected 

with HIV and 3 (2%) had an Orthotopic Liver Transplant 

(OLT). Prior to initiation of therapy; mean MELD score was 9 

(SD 4.96, range 6-29) and 53 (37%) had a Fibroscan score of 

> 12.5 kpa. The majority (91,64%,) had genotype 1 infection 

and 75 (53%) were treatment naïve. Patients who had failed 

first generation protease inhibitors were included. Opiate substitution 

therapy was prescribed in 42 (30%) patients. To date 

101/109 (93%) planned patient episodes are complete and 

33 remain on treatment. Of the 8 patient episodes where treatment 

was incomplete, 2 patients receiving peg/simeprevir/ 

ribavirin switched to sofosbuvir/ledipasvir due to intolerable 

IFN related side effects however completed the all oral regimen, 

2 receiving daclatasvir/sofosbuvir died from end stage 

liver disease and 2 developed non resectable/transplantable. 

Treatment was incomplete in 2 patients who received sofosbuvir/ledipasvir 

+ ribavirin, one due to suicidal ideation after 1 

week, the other developed renal failure after 3 weeks. The new 

treatment regimens are well tolerated; to date 93% of patients 

have completed their planned course; the majority who failed 

to complete had advanced liver disease. No-one has been lost 

to follow up. We would hope that SVR rates will be comparable 

to those reported in clinical trials. 

TABLE 1 

Disclosures: 

The following authors have nothing to disclose: Pauline Dundas, Shirley English, 

Lorna Bailey, Ashis Mukhopadhya, Balasubramaniam Vijayan, Andrew Fraser, 

Lindsay McLeman 

1105 

Asian Patients with Genotype 1 HCV Infection Achieve 

99% Sustained Virologic Response with 12 Weeks of 

Ledipasvir/Sofosbuvir Single Tablet Regimen: Integrated 

analysis of Phase 3 Multicenter Studies 

Masashi Mizokami 2 , Wan-Long Chuang 3 , Kwang-Hyub Han 4 , 

Young-Suk Lim 5 , Jenny C. Yang 1 , Shampa De-Oertel 1 , Bing 

Gao 1 , Hongmei Mo 1 , Phillip S. Pang 1 , Steven J. Knox 1 , John G. 

McHutchison 1 , Masao Omata 6 , Jia-Horng Kao 7 ; 1 Gilead Sciences, 

Foster City, CA; 2 National Center for Global Health and Medicine, 

Tokyo, Japan; 3 Kaohsiung Medical University, Kaohsiung, Taiwan; 

4 Yonsei University College of Medicine, Seoul, Korea (the Republic 

of); 5 University of Ulsan College of Medicine, Asan Medical Center, 

Seoul, Korea (the Republic of); 6 Yamanashi Prefectural Hospital 

Organization, Yamanashi, Japan; 7 National Taiwan University 

College of Medicine, Taipei, Taiwan 

Introduction: Similar to the United States and Europe, the majority 

of patients with chronic HCV infection in Japan, Korea, and 

Taiwan are infected with HCV genotype (GT) 1. However, 

important differences in viral and host characteristics exist 

between the infected populations in these regions, including 

age, BMI, IL28B genotype and HCV GT1 subtype. The aim of 

this integrated analysis is to evaluate the efficacy and safety 

of LDV/SOF in a large cohort of Asian patients with chronic 

GT1 HCV infection. Methods: This analysis combines data 

from subjects enrolled in two Phase 3 trials: GS-US-337-0113 

(Japan) and GS-US-337-0131 (Korea and Taiwan) evaluating 

12 weeks of LDV/SOF (90mg/400mg) in treatment-naïve and 

treatment-experienced adults with chronic GT1 HCV infection. 

Eligibility criteria: no upper age limit restriction, inclusion of 

subjects with compensated cirrhosis, platelet count ≥50,000/ 

mL and no restriction on neutrophil values. HCV NS5A and 

NS5B resistance associated variants (RAV) were evaluated by 

deep sequencing (cutoff of 1%). The primary efficacy endpoint 

was SVR12. Results: Overall, 349 subjects were enrolled in 

Korea, Taiwan, and Japan, 67 (19%) had cirrhosis. The majority 

were female (58%), treatment-experienced (51%), GT1b 

infected (94%), and IL28B CC (62%). The mean age (range) 

was 57 (18-80) years old, BMI 24 (17-38) kg/m 2 , and HCV 

RNA was 6.6 (3.7-7.6) log 10 

IU/mL. HCV NS5A RAVs were 

detected in 23% (80/343) of subjects at baseline. The overall 

SVR12 rate was 99% (346/349); 2 subjects relapsed and 1 

subject prematurely discontinued treatment. All treatment-experienced 

subjects with cirrhosis (45/45) achieved SVR12. 

NS5A RAVs were detected at the time of relapse but no NS5B 

RAVs were detected. Serious AE and treatment discontinuations 

were rare (





Sciences 

Masao Omata - Advisory Committees or Review Panels: Boehringer Ingelheim; 

Speaking and Teaching: Otsuka Pharmaceutical, Bayer 

The following authors have nothing to disclose: Masashi Mizokami, Kwang-Hyub 

Han, Jia-Horng Kao 

1106 

Efficacy, Change in MELD Score, and Safety by Baseline 

MELD Score in Patients With Compensated Cirrhosis 

Receiving Ombitasvir/Paritaprevir/r and Dasabuvir Plus 

Ribavirin in the Phase 3 TURQUOISE-II Trial 

Ira M. Jacobson 1 , Tania M. Welzel 2 , Hugo E. Vargas 3 , Marcos 

C. Pedrosa 4 , Norah Terrault 5 , Douglas Dieterich 6 , Fredric D. 

Gordon 7 , Kris V. Kowdley 8 , Guy Neff 4 , Ran Liu 4 , Juan-Carlos 

Lopez-Talavera 4 , Stefan Zeuzem 2 ; 1 Weill Cornell Medical College, 

New York, NY; 2 J.W. Goethe University, Frankfurt, Germany; 

3 Mayo Clinic Arizona, Phoenix, AZ; 4 AbbVie Inc., North Chicago, 

IL; 5 University of California San Francisco, San Francisco, 

CA; 6 Mount Sinai School of Medicine, Icahn School of Medicine 

at Mount Sinai, New York, NY; 7 Lahey Hospital & Medical Center, 

Burlington, MA; 8 Liver Care Network, Swedish Medical Center, 

Seattle, WA 

Purpose: Curing HCV infection reduces the risk of complications 

such as liver decompensation and hepatocellular carcinoma. 

The 3 direct-acting antiviral (3D) regimens of co-formulated 

ombitasvir/paritaprevir/r (paritaprevir identified by AbbVie 

and Enanta, dosed with ritonavir [r]) and dasabuvir +/- ribavirin 

(RBV) have achieved high SVR12 rates among adults with 

chronic genotype 1 HCV, including those with compensated 

cirrhosis. Model for end-stage liver disease (MELD) scores 

assess liver disease severity. In this analysis, we evaluated 

efficacy and safety of 3D+RBV and changes in MELD score by 

baseline MELD score in the phase 3 TURQUOISE-II trial. Methods: 

Treatment-naïve or peginterferon/RBV treatment-experienced 

patients with compensated cirrhosis were randomized 

to receive either 12 or 24 weeks of 3D+RBV in TURQUOISE-II. 

SVR12, change in MELD score, and adverse event rates are 

reported for subgroups of patients with baseline MELD scores 

of 6-9, 10-13, and >14. Results: 380 patients were randomized 

and received either 12 or 24 weeks of study drug. SVR12 

rates were 94.2% (322/342), 93.9% (31/33), and 80.0% 

(4/5) in patients with baseline MELD scores of 6-9, 10-13, and 

>14, respectively. The 1 patient with MELD >14 not achieving 

SVR12 discontinued study drug prematurely. Changes in MELD 

score in patients who achieved SVR12 is in the table. Rate of 

serious adverse events was 5.3% (18/342), 6.1% (2/33), 

and 0% (0/5) for the 6-9, 10-13, and >14 subgroups, respectively. 

Rate of discontinuation due to adverse events was 2.0% 

(7/342), 0% (0/33), and 20.0% (1/5) for the 6-9, 10-13, and 

>14 subgroups, respectively. Conclusions: In TURQUOISE-II, 

3D+RBV demonstrated high SVR12 rates and favorable safety 

in cirrhotic patients regardless of baseline MELD score. While 

this analysis is limited by the small number of patients, the data 

suggest improvements in MELD scores in patients with higher 

baseline MELD scores. 

Table. Changes in MELD score for patients who achieved SVR12. 

*PTW48 data were not available for all patients who achieved 

SVR12. Only data for those patients with data at PTW48 is 

shown. PTW, post-treatment week. 

Disclosures: 

Ira M. Jacobson - Consulting: AbbVie, Achillion, Alnylam, Bristol Myers Squibb, 

Enanta, Gilead, Janssen, Merck; Grant/Research Support: AbbVie, Bristol Myers 

Squibb, Gilead, Janssen, Merck, Tobira; Speaking and Teaching: AbbVie, Bristol 

Myers Squibb, Gilead, Janssen 








Fredric D. Gordon - Advisory Committees or Review Panels: Gilead, AbbVie; 

Grant/Research Support: BMS, Vertex, Gilead, AbbVie 




Gilead 


Juan-Carlos Lopez-Talavera - Employment: Abbvie; Stock Shareholder: BMS 



The following authors have nothing to disclose: Hugo E. Vargas, Marcos C. 

Pedrosa, Guy Neff 

1107 

Efficacy and safety of ombitasvir/paritaprevir/r and 

dasabuvir +/- ribavirin in HCV genotype 1-infected 

patients with a history of bleeding disorders: Results 

from phase 3 trials 

Giustino Parruti 1 , Guilherme Macedo 2 , Axel Baumgarten 3 , 

Frederik Nevens 4 , Jordan J. Feld 5 , Christophe Hezode 6 , Lois 

M. Larsen 7 , Nancy Shulman 7 , Regis A. Vilchez 7 , Heiner Wedemeyer 

8 ; 1 Ospedale Civile Spirito Santo, Pescara, Italy; 2 Centro 

Hospitalar de São João, Porto, Portugal; 3 Medical Center for Infectious 

Diseases, Berlin, Germany; 4 University Hospitals KU, Leuven, 

Belgium; 5 Toronto Centre for Liver Disease, University of Toronto, 

Toronto, ON, Canada; 6 Henri Mondor University Hospital, AP-HP, 

Université Paris-Est, Créteil, France; 7 AbbVie Inc., North Chicago, 

IL; 8 Medizinische Hochschule Hannover, Hannover, Germany 

Background: HCV infection has been prevalent in patients with 

bleeding disorders due to the high frequency of contamination 

of blood-derived clotting factor products prior to the 1990s. 

The 3 direct-acting antiviral (3D) regimens of co-formulated 

ombitasvir/paritaprevir/ritonavir (OBV/PTV/r, PTV identified 

by AbbVie and Enanta) and dasabuvir (DSV) +/- ribavirin 

(RBV) have achieved high SVR12 rates among adults with 

chronic genotype 1 HCV including those with compensated 

cirrhosis. In this analysis, we examined the efficacy and safety 

of 3D+/-RBV in patients with a history of bleeding disorders in 

phase 3 trials. Methods: Treatment-naïve or peginterferon/RBV 

treatment-experienced, cirrhotic or non-cirrhotic HCV genotype 

1-infected patients enrolled in phase 3 trials and received 

3D+/-weight-based RBV. The efficacy (SVR12 rate) and safety 

for patients with a history of bleeding disorders is reported. 

Results: Thirty-five patients with a history of bleeding disorders 

(29 of whom had hemophilia) were enrolled and received 

active study drug (32 patients, 3D+RBV; 3 patients, 3D). Baseline 

characteristics and efficacy results are in the table. Fifteen


patients were peginterferon/RBV treatment-experienced and 9 

patients were cirrhotic. SVR12 rate was 100% (35/35, 95% 

CI: 90.1%, 100%) among patients with a history of bleeding 

disorders. Among the 35 patients, there were no discontinuations 

due to adverse events. The most common adverse events 

were nausea, fatigue, and headache. One patient (2.9%) had 

a decline in hemoglobin to


1109 

Lower 25-OH Vitamin D Levels are Associated With 

Advanced Fibrosis in Chronic Hepatitis C Infection but 

Pretreatment or Change in Level with Directing Acting 

Antiviral Therapy Do Not Predict Sustained Virologic 

Response 

David W. Backstedt 1 , Mark Pedersen 2 , Bobby Kakati 1 , Myunghan 

Choi 4 , Anil B. Seetharam 3 ; 1 Gastroenterology, Banner University 

Medical Center, Phoenix, AZ; 2 Internal Medicine, Banner University 

Medical Center, Phoenix, AZ; 3 University of Arizona College 

of Medicine, Banner Transplant and Advanced Liver Disease Center, 

Phoenix, AZ; 4 Arizona State University College of Nursing and 

Health Innovation, Phoenix, AZ 

Rationale: Recent reports suggest association between low 

25-OH Vitamin D (Vit D) levels and advanced fibrosis in Hepatitis 

C Virus (HCV) infection. Evaluation of the influence and 

predictive value of pretreatment Vit D level on sustained virologic 

response (SVR) with direct acting antiviral (DAA) therapy 

is needed. Aim: Evaluate association of pre-treatment Vit 

D level with stage of fibrosis in an HCV infected cohort and 

assess change in level with DAA therapy. Secondary aim was 

to assess predictive value of pre-treatment or change in Vit D 

level on SVR12. Methods: Prospective study of chronic HCV 

patients initiated on DAA. Consecutively enrolled patients 

received: 1) pegylated interferon alfa 2a (IFN) + sofobuvir 

(SOF) + ribavirin (RBV); 2) SOF + RBV; 3) SOF + Simeprevir 

(SMV); and 4) Ledipasvir (LDV) + SOF; 5) LDV + SOF + RBV in 

accordance with AASLD guidelines. Vit D levels were assessed 

prior to initiation of treatment and at end of treatment response 

(ETR). Pearson correlation coefficients were assessed followed 

by repeated measures ANOVA to determine significant Vit D 

level changes over time controlling for covariates: age, gender, 

and BMI. Multivariate logistic regression was performed 

to identify predictors of SVR12. Results: 218 patients completing 

DAA regimen: IFN + SOF + RBV (n=37); SOF + RBV 

(n=117); SMV + SOF (n=51), LDV+SOF (n=11), LDV +SOF 

+RIBA (n=2). 61.5% genotype 1 and 56% cirrhosis. 64% 

treatment naïve and 36% treatment experienced (26% non-responders, 

10% relapsers). 98% (213/218) achieved ETR with 

79% (172/218) achieving SVR12 (19% relapsers). Frequency 

of low pre-treatment Vit D level (


1111 

Sofosbuvir and Ledipasvir for the Treatment of HCV 

GT-1, Cirrhotics and Non-Cirrhotics: Real-World Effectiveness 

Kirat Gill, Garrett Fante, Shirin Nafisi, Ann Moore, Ashley Hepner, 

Justin A. Reynolds, Karen Arendt, Yvette Cummings, Robert Gish, 

Richard A. Manch, Anita Kohli; Hepatology, St. Joseph’s Hospital 

and Medical Center, Creighton University School of Medicine, 

Phoenix, AZ 

The interferon and ribavirin (RBV) free combination of sofosbuvir 

(SOF) and ledipasvir (LDV), a hepatitis C nucleotide NS5B 

and N5A inhibitor respectively, have demonstrated high rates 

of sustained virologic response (SVR) in clinical trials of both 

treatment naïve and experienced patients, as well as cirrhotic 

and non-cirrhotic patients, with chronic hepatitis C (CHC). Real 

world non-clinical trial data for patients with CHC treated with 

SOF/LDV ± RBV is lacking. AIM: To evaluate the effectiveness 

of SOF/LDV for treatment of HCV genotype 1 (GT1) in a realworld 

patient population. METHODS: We performed a retrospective 

analysis of patients with CHC GT1 treated between 

October 2014-June 2015 (n=197) at a community based clinic 

within an academic medical center in Phoenix, AZ or by ECHO 

telemedicine to rural clinic sites. Demographics, CHC treatment 

regimens, liver fibrosis, virologic data and psychiatric 

history were collected throughout treatment and post-treatment 

from patient charts. RESULTS: Since October 2014, 197 GT1 

patients started treatment for CHC and are included in the 

current analysis. 81.7% of patients were Caucasian, 11.2% 

of patients were Hispanic and 1.5% Native American. 65% of 

patients were men, the mean age was 59 yrs. (range: 18-82 

yrs.) and mean HCV viral load 2,359,054 IU/mL (range: 

1,107-18,285,438 IU/mL). 75% of patients were GT 1a and 

24% were GT1b. 53.3% of patients had cirrhosis by imaging, 

Fibrosure or liver biopsy. 29.4% of patients had a concomitant 

psychiatric illness. 45.2% (n=89) of patients were previously 

treated. 50% (n=100) of patients were treated with the SOF/ 

LDV for 12 weeks, 31.0% (n=61) with SOF/LDV for 24 weeks, 

18.3% (n=36) with SOF/LDV for 8 weeks and 1.0% (n=2) 

treated with SOF/LDV/RBV for 12 weeks. Overall, 87.5% of 

patients treated with SOF/LDV for 12 weeks and who have 

reached the SVR12 timepoint (n= 16) have achieved SVR12, 

and 12.5% have relapsed (pending for 84 patients). SVR12 

data is pending for 60 patients treated with 24 weeks with 

SOF/LDV with one patient having relapsed. 100% of patients 

treated with SOF/LDV for 8 weeks and who have reached 

the SVR12 timepoint (n=10) have achieved SVR12 (pending 

for 26 patients). SVR12 is pending in the two patients treated 

with SOF/LDV/RBV. Full results of SVR 12 will be presented 

and stratified by cirrhosis and viral genotype. CONCLUSION: 

Treatment for hepatitis C GT1 using SOF/LDV based therapies 

has been well tolerated with high rates of SVR to date. SVR 

results in this real world population, enriched for patients with 

advanced liver disease, and treated by hepatologists as well as 

primary care providers, will be presented. 

Disclosures: 




Richard A. Manch - Speaking and Teaching: Gilead, AbbVie, Bayer, Salix, BMS 

The following authors have nothing to disclose: Kirat Gill, Garrett Fante, Shirin 

Nafisi, Ann Moore, Ashley Hepner, Justin A. Reynolds, Karen Arendt, Yvette 

Cummings, Anita Kohli 

1112 

Telaprevir in the Treatment of Acute HCV infection in 

HIV-infected Men: Final Results 

Daniel S. Fierer 2 , Douglas Dieterich 1 , Michael P. Mullen 2 , Andrea 

D. Branch 1 , Alison J. Uriel 1,6 , Damaris C. Carriero 1,7 , Wouter van 

Seggelen 2,3 , Rosanne M. Hijdra 2,3 , David Cassagnol 2,5 , Tristan 

Morey 2,4 ; 1 Liver Diseases, Icahn School of Medicine at Mount 

Sinai, New York, NY; 2 Infectious Diseases, Icahn School of Medicine 

at Mount Sinai, New York, NY; 3 Amsterdam Medical Center, 

Amsterdam, Netherlands; 4 James Cook University, Cairns, QLD, 

Australia; 5 Weill Cornell Medical College, New York, NY; 6 North 

Manchester General Hospital, Manchester, United Kingdom; 

7 Columbia University School of Medicine, New York, NY 

Background Treatment of HCV with pegylated interferon (pIFN) 

and ribavirin (RBV) is significantly more effective in the acute 

phase than in the chronic phase. Incorporating telaprevir (TVR) 

into the treatment of chronic HCV both improves the efficacy 

and shortens the duration of treatment, and our preliminary evidence 

suggested it can do the same in acute HCV. We present 

here the final results of our study of TVR + pIFN + RBV for acute 

HCV in HIV-infected men. Methods This is an IRB-approved, 

open-label, consecutive enrollment study of TVR (TID or BID) 

+ pIFN-α 180 μg/week + weight-based RBV for 12 weeks in 

HIV-infected men with acute genotype 1 HCV infection. Stopping 

rule was HCV VL > 1,000 IU/mL at week 4. Allowed 

ARVs were tenofovir+emtricitabine, efavirenz (with TVR dose 

adjustment), rilpivirine, atazanavir/ritonavir, and raltegravir. 

HCV VL was measured by transcription-mediated amplification 

(TMA, lower limit of detection 5 IU/mL). The comparator 

group was HIV-infected men with acute genotype 1 HCV either 

treated during the 3 years prior to the FDA approval of TVR or 

those ineligible for TVR. Results Thirty-six men in the TVR group 

and 50 men in the comparator group completed treatment and 

SVR 12 assessment. The two treatment groups did not differ in 

proportion of genotype 1a (84% vs 90%, respectively), IL28B 

CC (45% vs 42%, respectively), age, or race/ethnicity. Thirty-two 

(89%) achieved SVR 12 in the TVR group compared to 

32 (64%) in the comparator group (p = 0.01). Most (88%) in 

the TVR group received ≤12 weeks total treatment, while all 

in the comparator group received ≥24 weeks total treatment. 

TVR treatment resulted in faster VL kinetics among those who 

achieved SVR, with median time to HCV VL 75%. With the availability now of interferon-free regimens, 

though, efforts should move toward developing these less toxic 

regimens. Clinicians who do not have access to interferon-free 

regimens, however, should be alert to detect acute HCV infection 

in HIV-infected men to take advantage of this highly-effective 

TVR-based therapy. 

Disclosures: 




Michael P. Mullen - Advisory Committees or Review Panels: GILEAD; Speaking 

and Teaching: GILEAD 


Damaris C. Carriero - Advisory Committees or Review Panels: Abbvie, BMS; 

Consulting: Gilead; Speaking and Teaching: Gilead 

The following authors have nothing to disclose: Alison J. Uriel, Wouter van Seggelen, 

Rosanne M. Hijdra, David Cassagnol, Tristan Morey


1113 

Transient renal dysfunction during INF-free therapy in 

decompensated HCV cirrhosis patients 

Ivana Carey, Suman Verma, Anna Mrzljak, Aisling B. Considine, 

Sarah Knighton, Kath Oakes, Kate Childs, Matthew J. Bruce, Mary 

Horner, Abid Suddle, Kosh Agarwal; Institute of Liver Studies, 

Kings College School of Medicine at King’s College Hospital, London, 


Transient renal dysfunction occurs in 5-7% HCV patients 

treated with triple antiviral peg-IFN based therapy. Similarly 

Coilly at al reported transient decline in estimated glomerular 

filtration (eGFR) in cohort of post-transplant HCV patients 

treated with IFN-free (sofosbuvir (SOF) + daclatasvir (DCV) +/- 

RBV). On-treatment changes in specific renal markers cystatin 

C (glomerular function) and neutrophil gelatinase-associated 

lipocalin (NGAL) (tubular function), used in diagnosis of acute 

kidney injury, enhanced therapy delivery in patients on triple 

antiviral therapy. There are no data on cystatin C and NGAL 

levels changes during IFN-free therapy in HCV patients with 

decompensated cirrhosis (Child-Pugh B ≥7). Aim: Retrospective 

cross-sectional single centre study to investigate on-treatment 

changes in cystatin C and NGAL levels and correlate 

with changes in eGFR and presence/absence of proteinuria 

during IFN-free therapy for 12 weeks (SOF/ledipasvir+RBV or 

SOF+DCV+RBV) in HCV patients with decompensated cirrhosis. 

Patients: 52 patients with HCV decompesnated cirrhosis 

(median age 57years, 33 males, median MELD 11) underwent 

12 weeks of therapy with IFN-free DAA regimen (86% SOF/ 

ledipasvir+ RBV and 14% SOF+DCV+RBV) and completed 12 

weeks post-therapy follow-up. Material and Methods: Plasma 

levels of cystatin C, NGAL and renin at baseline, treatment 

week 4 (TW4), end of therapy (EoT), follow-up week 4 (FUW4) 

and follow-up week 12 (FUW12) were measured by ELISA 

and compared with eGFR and urinary protein creatinine ratio 

(uPCR) at the same time-points. Patients were stratified according 

to pre-existing renal risks (RR) (hypertension, diabetes, 

diuretics, tacrolimus). The results are presented as medians. 

Results: 50% patients had renal risk; 16 hypertension, 10 diabetes, 

26 therapy with diuretics and 6 received tacrolimus. 

At baseline 14% of patients had eGFR15 and these proportions did not change significantly 

during therapy and follow up. eGFR did not change 

significantly during therapy and was lower in RR than non-RR 

patients (74 vs. 93min/ml, p=0.02). Cystatin C and NGAL 

levels were higher in RR patients and increased during the first 

4 weeks of therapy in all patients (TW0 vs. TW4- cystatin C: 

1.46 vs. 1.55mg/l, p


1115 

Resistance-associated variants in NS5A region of hepatitis 

C virus are susceptible to interferon-based therapy 

Jun Itakura, Masayuki Kurosaki, Natsuko Nakakuki, Hitomi 

Takada, Nobuharu Tamaki, Yutaka Yasui, Shoko Suzuki, Kaoru 

Tsuchiya, Hiroyuki Nakanishi, Namiki Izumi; Division of Gastroenterology 

and Hepatology, Musashino Red Cross Hospital, Tokyo, 

Japan 

BACKGROUND & AIMS: Treatments with direct acting antivirals 

(DAAs) has become the standard of chronic hepatitis C treatment 

in Japan. But the presence of resistance-associated variants 

(RAVs) of hepatitis C virus (HCV) attenuates the efficacy of 

DAAs. The aim of this study was to characterize the susceptibility 

of RAVs to interferon-based therapy. METHODS: Twenty nine 

genotype 1b patients with detectable Y93H variant in NS5A 

region at baseline were enrolled and treated by a combination 

of simeprevir (NS3/4A inhibitor), pegylated interferon and 

ribavirin. The longitudinal changes of RAV were determined 

by direct sequencing during therapy and at breakthrough or 

relapse, and the longitudinal changes in the proportion of 

Y93H RAV were determined by deep sequencing. RESULTS: 

By direct sequencing, Y93H RAV became undetectable or its 

proportion decreased in 57% of patients with both Y93H and 

Y93wild type at baseline at an early time point during therapy 

when HCV RNA was still detectable. By deep sequencing, the 

proportion of Y93H RAV was decreased from 52.7% (5.8% – 

97.4%) at baseline to 29.7% (0.16% - 98.3%) within 7 days of 

initiation of treatment (p=0.023). The proportion of Y93H RAV 

against Y93 wild type was reduced in 21 of 29 cases (72.4%) 

and a marked reduction of more than 10% was observed in 

14 cases (48.7%). In 4 cases with breakthrough/relapse, the 

proportion of Y93H RAV decreased during therapy and at 

breakthrough/relapse but recovered to baseline 3 months after 

discontinuation of treatment. HCV RNA reduction was significantly 

greater for Y93H RAV (-3.65±1.3 logIU/mL/day) than 

the Y93wild type (-3.35±1.0 logIU/mL/day) (p 

Shuhei Hige, Itaru Ozeki, Masakatsu Yamaguchi, Mutuumi 

Kimura, Tomohiro Arakawa, Tomoaki Nakajima, Yasuaki Kuwata, 

Takahiro Sato, Takumi Ohmura, Joji Toyota, Yoshiyasu Karino; 

Hepatology, Sapporo-Kosei General Hospital, Sapporo, Japan 

Background/Aims: One of the most serious problems in the 

treatment of hepatitis C is the aging of hepatitis C patients 

who had been left behind without being treated with interferon-based 

therapy. Combination therapy with daclatasvir (DCV) 

and asunaprevir (ASV) is the only option for Japanese patients 

with genotype 1b chronic hepatitis C at the moment. However, 

the efficacy and safety of DCV/ASV combination therapy has 

not been fully elucidated especially for those who are in their 

late 70’s and 80’s. The aim of this study was to evaluate the 

efficacy and safety of DCV/ASV for senior patients including 

pharmacological analysis. Patients and Methods: Two 

hundred and ninety-three genotype 1 patients were treated 

with fixed daily dose of DCV (60mg) and ASV (200mg) for 

24 weeks. The patients were divided into 3 groups by age: 

Group I (


mononuclear cells were collected at baseline, treatment week 

4 (TW4), end-of treatment (EoT) and follow-up week 12 in all 

patients and flowcytometry was performed after multicolour 

staining for NK cells subsets (CD3, CD16, CD56, NKp30 and 

NKG2D). The frequency of NK cells subsets was compared 

between responders (R) (n=5) vs. relapsers (REL) (n=6) at different 

therapy time-points. Plasma IP10, IL10 and IL12 levels were 

measured by ELISA at same time-points. Results are presented 

as medians. Results: Baseline HCV RNA levels were similar 

between R and REL. Overall NK cells frequency (CD3-CD56+) 

was higher in R than REL at baseline (5.79% vs. 2.38%, 

p=0.02). The frequency of NK cells increased during therapy, 

but proportions remained higher in R than REL (7.5% vs. 4.2%). 

CD56bright cells subset was higher in R than REL at baseline 

(14.5% vs. 7.5%, p=0.01), but increased only in REL at EoT 

to 14.8% and dropped to pre-treatment levels (7.6%) at FU. 

The frequency of NKG2D was similar between R and REL at all 

time-points. NKp30+ cells increased during therapy only in REL 

than R (11.2% vs. 1.1%, p


ties and anemia did not affect SRV-12. A sub-group of patients 

over age 75, analyzed separately, showed SVR-12 of 100%, 

10% reported fatigue and 5.2% had diarrhea, with no hospitalizations 

during the treatment. Conclusion: Sofosbuvir based 

regimen is safe and effective in patients over the age of 65 

years including those with cirrhosis. 

Genotype (GT) 

Treatment naïve (TN) 

Treatment experienced (TE) 

Disclosures: 

The following authors have nothing to disclose: Beshoy T. Yanny, Amandeep K. 

Sahota 

1120 

Sofosbuvir and Ledipasvir/Sofosbuvir for the Treatment 

of Patients with Chronic Genotype 6 Hepatitis C Virus 

Infection: Integrated Analysis of Phase 2 and Phase 3 

Studies 

Edward J. Gane 1 , Wan-Long Chuang 2 , Tarek I. Hassanein 3 , 

Kris V. Kowdley 4 , Eric Lawitz 5 , Bing Gao 6 , Hongmei Mo 6 , Robert 

H. Hyland 6 , Jenny C. Yang 6 , Shampa De-Oertel 6 , Diana M. 

Brainard 6 , Steven J. Knox 6 , John G. McHutchison 6 , Ching-Lung 

Lai 7 , Henry Lik-Yuen Chan 8 ; 1 New Zealand Liver Transplant Unit, 

Auckland, New Zealand; 2 Kaohsiung Medical University, Kaohsiung, 

Taiwan; 3 Southern California Liver Centers, Coronado, CA; 

4 Swedish Medical Center, Seattle, WA; 5 The Texas Liver Institute, 

San Antonio, TX; 6 Gilead Sciences, Inc, Foster City, CA; 7 Queen 

Mary Hospital, Hong Kong, Hong Kong; 8 The Chinese University 

of Hong Kong, Hong Kong, Hong Kong 

Introduction: Chronic HCV infection is endemic in South East 

Asia with HCV genotype 6 (GT6) accounting for 18-49% of 

those infected. In the US and Canada, nearly one-third of immigrants 

from Southeast Asia with HCV are infected with GT6. 

Few studies have examined the efficacy and safety of direct 

acting antiviral (DAA) regimens in GT6 infected patients. The 

aim of this integrated analysis was to characterize the efficacy 

and safety of sofosbuvir (SOF)-based regimens in patients with 

chronic GT6 HCV infection. Methods: GT6 infected subjects 

were identified in 5 studies (ATOMIC, NEUTRINO, GS-US- 

334-0115, ELECTRON2, GS-US-337-0131) and are included 

in this analysis. Treatment-naïve or treatment-experienced 

patients received SOF+RBV±Peg-IFNα or ledipasvir (LDV)/SOF 

for 12-24 weeks. The primary efficacy endpoint in all studies 

was SVR12. Results: A total of 52 subjects with GT6 HCV infection 

were identified. The majority were treatment-naïve (94%), 

Asian (81%), male (58%), and had IL28B CC alleles (81%). 

The mean age was 50 years (range 26-76) and 10% had cirrhosis. 

GT6 subtypes included 6a, 6a/b, 6c-1, 6e, 6g, 6j, 6l, 

6m, 6o, 6p, 6q, and 6r. The table below presents SVR12 by 

regimen. One subject in ELECTRON2 withdrew consent after 

receiving 8 weeks of LDV/SOF and relapsed with the emergent 

NS5B RAV S282T. All remaining 51 patients achieved 

SVR12, including 100% (3/3) experienced and 100% (5/5) 

cirrhotics. Conclusions: SOF+RBV±Peg-IFNα and LDV/SOF 

regimens are well-tolerated and highly effective in patients with 

chronic GT6 HCV infection including those who are treatment 

experienced and have compensated cirrhosis. These regimens 

provide multiple therapeutic options for consideration when 

evaluating optimal therapy for individual patients with chronic 

GT6 HCV infection. 

SVR12 Rates in GT6 HCV Patients 

1ATOMIC (P7977-0724), NEUTRINO (GS-US-334-0110); 2GS- 

US-334-0115; 3ELECTRON2 (GS-US-337-0122), GS-US-337- 

0131 

Disclosures: 
















Gilead 








Bing Gao - Employment: Gilead; Stock Shareholder: Gilead 



Sciences, Inc 


Shampa De-Oertel - Employment: Gilead Sciences, Inc; Stock Shareholder: Gilead 

Sciences, Inc 


Sciences 



Sciences 







1121 

Real Life Experience with Sofosbuvir and Ledipasvir 

Fixed Dose Regimen in the Multiethnic Cohort of Patients 

with Chronic Hepatitis C 

Marina Roytman 2,1 , Alister L. Tang 1 , Christina Wu 1 , Joy I. 

Piotrowski 1 , Leena K. Hong 2 , Ruby Trujillo 2 , Leslie Huddleston 2 , 

Isabel Hung Wan 2 , Peter Poerzgen 2 , Sumodh Kalathil 2 , Naoky 

CS C. Tsai 2,1 ; 1 Medicine, University of Hawaii at Manoa, John A. 

Burns School of Medicine, Honolulu, HI; 2 Queen’s Liver Center, 

Honolulu, HI 

Purpose/Background: ION-1-3 trials have demonstrated high 

SVRs and favorable side effect profiles in the treatment of GT1 

HCV patients. However, reports of regimen experiences in 

the real life clinical setting are limited. This study investigates 

the safety and efficacy of Ledipasvir (LDV)- Sofosbuvir (SOF) 

regimen in the multiethnic patient cohort, including patients 

with decompensated cirrhosis, in a real life clinical setting. 

Methods: Retrospective review of 154 patients with genotype 

1 (GT1) chronic hepatitis C (HCV) infection treated with a


fixed dose Ledipasvir (LDV) and Sofosbuvir (SOF) (90/400mg) 

regimen November 2014- May 2015 at Hawaii’s single tertiary 

referral center. We collected data on demographics, 

adverse events, laboratory values, and SVR (sustained virological 

response). Statistical analysis was performed with R 

version 3.1.0. Results: Baseline characteristics: 42% cirrhotic 

(23.8% of those Child-Pugh Class B/C), 47.7% Caucasian, 

30.1% Asian, 9.2% Pacific Islander, 65.3% male, mean age 

60 ±10.5, mean BMI 27.1 ±5.8, 11% diabetic, 60.1% GT1a. 

Interim analysis data are presented. Viral load was negative 

in 100% of patients at the end of treatment (EOT). Main side 

effects: headache 14.4%, weight gain 7.8%, nausea 5.9%, 

and diarrhea 3.9%. Headache was more frequent in cirrhotics 

versus non-cirrhotics (19 vs 11%). In patients with baseline 

abnormal ALT (>30 U/L), 84/99 showed normal ALT levels 

(15 IU/cc) at 4 weeks post treatment. 

Conclusions: 12-week treatment was highly effective and well 

tolerated in a multiethnic cohort of patients including patients 

with advanced liver disease. Interim analysis indicates SVR 

rates comparable to ION trials despite higher proportion (42%) 

of cirrhotics in our cohort. Early improvement in albumin levels 

in cirrhotic patients may indicate improvement in synthetic function. 

Complete data on SVR4 and SVR12 will be available on 

all patients by October 2015. 

reduced toxicities, the new Direct Acting Antivirals (DAA) could 

be attractive therapeutic options for HCV treatment after KT. 

The approach of safe HCV treatment post KT, could allow the 

use of organs from HCV +ve donors, thus reducing organ discard 

rates the wait-list time for KT. Methods: 15 HCV +ve KT 

recipients were prospectively enrolled in the study at the time 

of initiation of DAA. We report the data from 8 recipients who 

had completed HCV treatment till now. Results: From the 8 

adult (Mean age 58.5 years; Male: Female=6:2) KT recipients, 

4 received HCV +ve grafts. 2 completed Sofosbuvir 400 

mg and Simeprevir 150 mg daily combination and 6 completed 

Sofosbuvir/Ledipasvir 150mg/90 mg daily 12 weeks 

regimen. All achieved End Treatment Response. Data is being 

collected for Sustained Virologic Responses (SVR). There were 

no episodes of graft rejection and none required modification 

in immunosuppression. There were no adverse events requiring 

cessation of therapy. All KT recipients had stable renal and 

liver function during and after the completion of therapy. Conclusion: 

In our cohort of KT recipients, DAAs showed excellent 

safety profile and good virologic response. 

Study population characteristics and treatment responses of each 

subject. 

(KT) Kidney Transplant 

(IFN) Interferon 

(R) Ribavirin 

(PI) Protease Inhibitor 

(T) Tacrolimus 

(M) Mycophenolate 

(P) Prednisone 

Disclosures: 

Marina Roytman - Advisory Committees or Review Panels: Gilead; Speaking and 

Teaching: Gilead 

Leena K. Hong - Advisory Committees or Review Panels: Gilead Sciences; Speaking 

and Teaching: Gilead Sciences, Abbvie 

Naoky CS C. Tsai - Advisory Committees or Review Panels: Bristol-Myers Squibb, 

Gilead, AbbVie; Consulting: BMS, Gilead; Grant/Research Support: BMS, 

Gilead, AbbVie; Speaking and Teaching: Bristol-Myers Squibb, Gilead, Salix, 

Bayer, AbbVie 

The following authors have nothing to disclose: Alister L. Tang, Christina Wu, Joy 

I. Piotrowski, Ruby Trujillo, Leslie Huddleston, Isabel Hung Wan, Peter Poerzgen, 

Sumodh Kalathil 

(Sof) Sofosbuvir 

(Sim) Simeprevir 

(Led) Ledipasvir 

(EOT) End of Treatment 

(SVR) Sustained Virologic Response 

1122 

Treatment of Chronic Hepatitis C Infection in Kidney 

Transplant Recipients with Direct Acting Antiviral Medications 

– Initial Experience 

Sushrut Trakroo 1 , Sirish Sanaka 1 , Hawah Musa 1 , Mohammed 

Eyad Yaseen Alsabbagh 2 , Mythili Ghanta 1 , Swati Rao 1 , Lee Peng 2 , 

Antonio Di Carlo 1 , Abdullah M. Al-Osaimi 2 , Kamran Qureshi 2 ; 

1 Temple University Hospital, Philadelphia, PA; 2 Gastroenterology 

and Hepatology, Temple University School of Medicine, Philadelphia, 

PA 

Background: Chronic Hepatitis C infection (HCV) is the leading 

cause of liver disease after Kidney Transplant (KT). HCV positive 

(+ve) KT recipients have worse overall and graft survival 

compared with HCV negative KT recipients. Treatment of HCV 

before KT with Interferon (IFN) based therapies showed poor 

response rates and tolerability. IFN use is relatively contraindicated 

after KT due to increased risk of allograft rejection 

and organ failure. Because of the shown greater efficacy and 

Disclosures: 

Abdullah M. Al-Osaimi - Advisory Committees or Review Panels: Abbvie, Gilead, 

Intercept Pharmaceuticals; Grant/Research Support: Chronic Liver Disease 

Foundation (CLDF), Beckman Coulter Inc., Conatus Pharmaceuticals, BioPharma 

Alliance, Bayer HealthCare Pharmaceuticals, Inc., Vital Therapies Inc., Roche 

Molecular Systems, Inc., Abbvie


The following authors have nothing to disclose: Sushrut Trakroo, Sirish Sanaka, 

Hawah Musa, Mohammed Eyad Yaseen Alsabbagh, Mythili Ghanta, Swati Rao, 

Lee Peng, Antonio Di Carlo, Kamran Qureshi 

1123 

Retreatment with sofosbuvir and simeprevir of patients 

who previously failed on an HCV NS5A inhibitor–containing 

regimen 

Christophe Hezode 1,2 , Stephane Chevaliez 3,2 , Giovanna 

Scoazec 1 , Alexandre Soulier 3,2 , Anne Varaut 1 , Magali Bouvier-Alias 

3,2 , Isaac Ruiz 3,2 , Ariane Mallat 1,2 , Cyrille Feray 1,2 , 

Jean-Michel Pawlotsky 3,2 ; 1 Hepatologie, Hospital Henri Mondor, 

Creteil, France; 2 INSERM U955, Creteil, France; 3 Virology, Henri 

Mondor Hospital, Creteil, France 

Background: Failure to achieve SVR with IFN-free regimens 

based on HCV DAAs is associated with the selection of resistant 

HCV variants. In contrast to variants resistant to PIs that 

are progressively overgrown by wild-type virus after treatment 

failure, NS5A RAVs persist for years as the dominant species 

in the majority of patients. In spite of the lack of data, recent 

guidelines recommended that patients exposed to NS5A inhibitors 

be retreated with a combination of SOF and SMV to avoid 

cross-resistance with NS5A inhibitors. Materials & Methods: 16 

patients (mean age: 54 yrs) infected with GT 1 (1a: n=11; 1b: 

n=3) or 4 (n=2) with compensated disease and advanced liver 

fibrosis (fibroscan: 9.6-70 kPa; cirrhosis n=9) who failed to 

achieve SVR after receiving DCV/PegIFN/RBV (n=13) DCV/ 

ASV/PegIFN/RBV (n=3), were included. They were retreated 

for 12 weeks with SOF/SMV, without RBV. HCV RNA levels 

were measured with Abbott RealTime Assay (LLOQ/LOD: 12 

IU/mL). HCV resistance was assessed at retreatment baseline 

by means of population sequencing home made methods targeting 

the NS3 and NS5A coding regions. Results: The median 

baseline HCV RNA level at retreatment was 1.38 x 10 6 IU/mL 

(>800,000 IU/mL: n=14). No patient discontinued treatment 

due to an adverse event or for virologic failure. Among the 16 

patients, 10 had a rapid virologic response (HCV RNA





The following authors have nothing to disclose: Ju Dong Yang, Bashar A. Aqel, 

Kymberly D. Watt, Hugo E. Vargas, Andrew P. Keaveny, Michael D. Leise 

1125 

Combination therapy with daclatasvir and asunaprevir 

in cirrhotic patients with hepatitis C virus genotype 1b: 

On-treatment efficacy and adverse effects 

Akihiro Tamori, Masaru Enomoto, Hoang Hai, Yuga Teranishi, 

Hiroyuki Motoyama, Ritsuzo Kozuka, Etsushi Kawamura, Atsushi 

Hagihara, Sawako K. Uchida, Hiroyasu Morikawa, Yoshiki 

Murakami, Norifumi Kawada; Hepatology, Osaka City University 

Graduate School of Medicine, Osaka, Japan 

Background and Aim: Combination therapy with daclatasvir 

(DCV; NS5A inhibitor) and asunaprevir (ASV; second-generation 

HCV-NS3/NS4A protease inhibitor) was approved for 

patients with HCV genotype 1 in Japan since September 2014. 

Now, elderly patients and those with advanced hepatic fibrosis 

including chronic liver cancer were administered IFN-free therapy. 

Our objective was to assess the efficacy and tolerability 

of DCV/ASV combination therapy in patients with hepatic cirrhosis. 

Methods: In total, 140 consecutive patients with HCV 

1b initiating DCV/ASV therapy were enrolled. The cohort comprised 

49 patients with compensated cirrhosis and 91 patients 

without cirrhosis (62 males and 78 females; median age, 71 

years; 6 patients were >80 years old). NS5A resistance-associated 

variants (RAV) were examined using direct sequencing. 

The patients were treated with 60 mg of DCV once daily and 

100 mg of ASV twice per day for 24 weeks. Clinical, biological, 

and virological data, including adverse effects, were 

recorded at baseline and during follow-up. Results: Only 10 

(7%) patients had L31M or Y93H RAVs. There was no statistically 

significant difference in age, sex, IL28B genotypes, HCV 

viral load at baseline, ALT level, creatinine level, or NS5A 

RAVs between patients with and without cirrhosis. On the other 

hand, those with cirrhosis showed significantly lower levels of 

platelets, white blood cells, and hemoglobin and higher levels 

of alpha fetoprotein. The rapid viral response rate (HCV-RNA < 

25 IU/ml at week 4) was the same between patients with and 

without cirrhosis (80% and 81%, respectively). Now, DCV/ 

ASV therapy was completed in 67 patients. One of 27 patients 

with cirrhosis, and two of 40 patients without cirrhosis who 

did not have NS5A RAVs at baseline developed viral breakthrough. 

The rate of SVR4 was 94% (16/17) in patients with 

cirrhosis and 90% (19/21) in patients without cirrhosis. Grade 

3/4 complications frequently occurred in 22% of patients with 

cirrhosis, of whom one had an elevated ALT level, two progressed 

to decompensated cirrhosis without ALT elevation, 

one developed HCC, one developed interstitial pneumonia, 

one had severe bronchitis, one had arterial fibrillation, three 

had gastrointestinal bleeding, and two developed edema. Of 

the patients without cirrhosis, ALT elevation was observed in 

two patients, and HCC developed in one patient. Conclusion: 

DCV/ASV therapy achieved a high anti-HCV effect in patients 

both with and without cirrhosis. However, careful management 

is necessary in patients with cirrhosis. 

Disclosures: 



The following authors have nothing to disclose: Akihiro Tamori, Masaru Enomoto, 

Hoang Hai, Yuga Teranishi, Hiroyuki Motoyama, Ritsuzo Kozuka, Etsushi 

Kawamura, Atsushi Hagihara, Sawako K. Uchida, Hiroyasu Morikawa, Yoshiki 

Murakami 

1126 

Safety and Efficacy of All-Oral Regimens for Hepatitis C 

in Cirrhotic Patients with Thalassemia 

Emmanouil Sinakos 1 , Dimitrios Kountouras 2,3 , Ioannis Koskinas 4 , 

Stylianos Karatapanis 5 , Alexandra Kourakli 6 , Efthimia Vlachaki 7 , 

Ioannis Goulis 1 , Antonios Kattamis 8 , Konstantinos Maragkos 9 , 

Fotini Petropoulou 10 , Barbara Toli 3 , Evangelos Akriviadis 1 , George 

V. Papatheodoridis 2 ; 1 4th Medical Department, Hippokratio Hospital, 

Aristotle University of Thessaloniki, Thessaloniki, Greece; 

2 Department of Gastroenterology, Laiko Hospital, University of Athens, 

Athens, Greece; 3 Mitera Hospital, Athens, Athens, Greece; 

4 2nd Medical Department, Hippokratio Hospital, University of Athens, 

Athens, Greece; 5 Department of Internal Medicine, Hospital 

of Rhodes, Rhodes, Greece; 6 Thalassemia Unit, Hospital of Patra, 

Patra, Greece; 7 Thalassemia Unit, Aristotle University of Thessaloniki, 

Thessaloniki, Greece; 8 Agia Sofia Children Hospital, University 

of Athens, Athens, Greece; 9 Thalassemia Unit, Hippokratio Hospital, 

Athens, Athens, Greece; 10 Thalassemia Unit, General Hospital, 

Athens, Athens, Greece 

Background-Aims: Thalassemia syndromes have been traditionally 

associated with high hepatitis C prevalence rates due 

to the regular blood transfusion requirements. Treatment with 

interferon and ribavirin usually leads to anemia, poor tolerance 

and subsequently low clearance rates, thus often failing to halt 

progression to cirrhosis. Although current guidelines suggest 

that interferon-free and ribavirin-free regimens should be used 

in these patients, no data have been yet presented. The aim 

of this study was to assess the safety and efficacy of all-oral 

regimens against hepatitis C virus in cirrhotic patients with thalassemia. 

Patients-Methods: Data on patients with thalassemia 

treated for hepatitis C in everyday clinical practice were analyzed. 

All patients were exposed to regular blood transfusions 

and chelation therapy. The predominant chelation therapy was 

combination of deferoxamine and deferiprone. The stage of 

liver disease was assessed histologically or with transient elastography. 

Three different regimens were used: sofosbuvir+ribavirin: 

n=3 (10%) for 24 weeks, sofosbuvir+simeprevir: n=13 

(43%) for 12 weeks and sofosbuvir+daclatasvir: n=14 (47%) 

for 12 weeks. Monitoring for side effects was performed every 

month during treatment and as needed after treatment. Results: 

30 patients with mean age of 45±5 years were included. All 

patients had compensated cirrhosis, except for one who had 

F3 stage. 25 patients (83%) were non-responders to previous 

courses (up to 5), either of (peg-)interferon and ribavirin (n= 

17), or interferon monotherapy (n=8). The genotype distribution 

was as follows: G1a: n=5 (17%), G1b: n=12 (40%), G2: 

n=1 (3%), G3: n=6 (20%), G4: n=6 (20%). Baseline laboratory 

values were: Hb=10.2±0.9 g/dL, serum ferritin=593±364 

ng/mL, AST=98±51 IU/mL, ALT=104±47 IU/mL, serum albumin=3.7±0.5 

g/dL, total bilirubin=2.2±1.5 mg/dL, HCV 

RNA=1.5±1.8x10 6 IU/ml. The mean liver magnetic resonance 

imaging Τ2* was 19.6±11.7 msec (data on 14 patients). At 

the time of this report 10 patients have completed treatment. 

No early discontinuation, increase in blood transfusion requirements 

or cardiac arrhythmia was reported. 7/8 (87%) patients 

evaluated have reached SVR12. Two patients with SVR12 had 

received sofosbuvir+ribavirin (one with G1b and one with G2) 

and five had received sofosbuvir+simeprevir (two with G1a 

and three with G1b). The only failure was observed in a patient 

with G1b who received sofosbuvir+ribavirin for 24 weeks. 

Conclusions: Treatment of hepatitis C with all oral regimens in 

cirrhotic patients with thalassemia is safe. Preliminary efficacy 

results are promising and seem not to differ from published SVR 

rates in other populations.


Disclosures: 

Emmanouil Sinakos - Advisory Committees or Review Panels: AbbVie; Speaking 

and Teaching: AbbVie, Bristol-Myers Squibb, Gilead, Janssen 

Ioannis Goulis - Consulting: Gilead Sciences, Abbvie, Janssen-Cilag, Janssen-Cilag, 

BMS; Grant/Research Support: BMS; Speaking and Teaching: BMS, Gilead 

Sciences, Janssen-Cilag 






The following authors have nothing to disclose: Dimitrios Kountouras, Ioannis 

Koskinas, Stylianos Karatapanis, Alexandra Kourakli, Efthimia Vlachaki, Antonios 

Kattamis, Konstantinos Maragkos, Fotini Petropoulou, Barbara Toli, Evangelos 

Akriviadis 

1127 

NS3 Q80K Polymorphism in Viral Isolates from Liver 

Tissues and Serum Samples of Naïve HCV Genotype 1a 

Patients. 

Deborah D’Aliberti 2 , Irene Cacciola 2 , Salvatore Benfatto 1 , Federica 

Mannino 1 , Roberto Filomia 2 , Concetta Beninati 1 , Giovanni 

Raimondo 2 , Teresa Pollicino 1 ; 1 Pediatric, Gynecological, Microbiological 

and Biomedical Sciences, University Hospital of Messina, 

Messina, Italy; 2 Clinical and Experimental Medicine, University 

Hospital of Messina, Messina, Italy 

Background and Aim – NS3 Q80K is a polymorphism commonly 

detected in genotype (G) 1a HCV strains. It is associated 

with a reduced antiviral activity of HCV proteinase inhibitors 

(PIs) in vitro and its emergence is generally considered a cause 

of response failure to Simeprevir as well as to other PIs in 

HCV-G1a patients. Several studies have evaluated the prevalence 

of Q80K polymorphism in HCV isolates from treatment 

naïve patients of different geographic areas. However, the 

data available concern only circulating viral isolates, whereas 

there is no information about the variability of HCV strains in 

the liver where the viral replication occurs and the HCV quasispecies 

diversity reaches the highest level of complexity. Aim of 

our study was to investigate the presence of Q80K substitution 

in HCV-G1a isolates from liver tissues and serum samples of 

patients from south of Italy, naïve to any antiviral treatment. 

Patients and Methods – HCV NS3 protease sequences of HCV 

isolates from paired serum and liver biopsy samples of 11 naïve 

patients with HCV-G1a chronic infection, and from serum samples 

of additional 20 naïve HCV-G1a infected patients were 

analysed by population sequencing and ultra-deep pyrosequencing 

(UDPS) (mutant detection sensitivity 1%; >3000 

sequences/patient). Results – The Q80K substitution was found 

in 8/11 liver tissues (72.7%; in 7 present at levels between 1% 

and 10%, and in 1 at level ≈97% of the virus quasispecies) and 

in 5/11 corresponding serum samples (45.4%; in 4 present at 

level ≈1%, and in 1 at level ≈97% of the virus quasispecies) by 

UDPS. Of the additional 20 sera examined, 13 (65%) showed 

the Q80K substitution by UDPS (in 11 present at level ≈1%, 

in 1 present at level ≈20%, and in 1 present at level ≈99% of 

the virus quasispecies). On the whole, the Q80K substitution 

was detected in 18/31 sera (58%) analysed. By population 

sequencing, the Q80K substitution was detected only in samples 

from 2 of the 31 patients (6.4%) analyzed. In particular, 

the Q80K substitution was found in the paired liver and serum 

samples from 1 patient and in the serum sample from another 

patient with no liver specimen available (in all these 3 samples 

the Q80K was present in 97-99% of the entire viral populations 

by UDPS). Conclusions – HCV-G1a strains carrying 

Q80K polymorphism are commonly present both in liver and 

in serum of infected patients. However, they are present as 

minor populations in almost all the cases, suggesting that the 

Q80K substitution does not confer fitness advantage at least to 

the HCV-G1a viral strains circulating in our geographic area. 

Disclosures: 

Giovanni Raimondo - Speaking and Teaching: BMS, Gilead, Roche, Merck, 

Janssen, Bayer, MSD 

Teresa Pollicino - Speaking and Teaching: Gilead, Roche, Janssen, BMS, MSD, 

Bayer, Abbvie 

The following authors have nothing to disclose: Deborah D’Aliberti, Irene Cacciola, 

Salvatore Benfatto, Federica Mannino, Roberto Filomia, Concetta Beninati 

1128 

Safety and Efficacy of Treatment with Daily Sofosbuvir 

400 mg + Ribavirin 200 mg for 24 Weeks in Genotype 

1 or 3 HCV-Infected Patients with Severe Renal Impairment 

Paul Martin 2 , Edward J. Gane 3 , Grisell Ortiz-Lasanta 4 , Lin Liu 1 , 

Karim Sajwani 1 , Brian Kirby 1 , Jill M. Denning 1 , Luisa M. Stamm 1 , 

Diana M. Brainard 1 , John G. McHutchison 1 , Eric Lawitz 5 , Stuart 

C. Gordon 6 , Richard A. Robson 7 ; 1 Gilead Sciences, Raleigh, NC; 

2 University of Miami, FL, USA, Miami, FL; 3 University of Auckland, 

Auckland, New Zealand; 4 Fundcion de Investigacion, San Juan, 

PR; 5 Texas Liver Institute, San Antonio, TX; 6 Henry Ford Health 

System, Detroit, MI; 7 Christchurch Clinical Studies Trust, Ltd, Christchurch, 

New Zealand 

Background: HCV-infected patients with severe renal impairment 

have had limited treatment options with suboptimal 

response rates and poor tolerability of IFN-based regimens. 

SOF 200 mg+RBV studied previously was safe and relatively 

well tolerated in patients with severe renal impairment, but SVR 

rates were low. The current study evaluated the safety, pharmacokinetics 

(PK) and efficacy of SOF 400mg+RBV for 24 weeks 

in genotype 1 or 3 HCV-infected patients with severe renal 

impairment. Methods: Treatment-naïve or experienced patients 

± cirrhosis and a creatinine clearance (CLcr) less than 30mL/ 

min (by Cockcroft-Gault equation), not on dialysis, received 

open-label treatment with SOF 400mg+RBV 200mg once daily 

for 24 wks. We examined on-treatment virologic response, PK 

and safety including echocardiograms read by a central reader 

pre-treatment and at Week 12 of therapy. Results: 10 patients (6 

GT1a, 2 GT1b, 2 GT3a) were enrolled and have been treated 

for 12-24 wks (median 16 wks): 8 male, 5 white, 4 black, 1 

Pacific Islander, mean age 58; 4 with cirrhosis, 7 treatment-experienced, 

5 IL28B genotype non-CC, mean baseline CLcr 

26.2 mL/min, and mean baseline hemoglobin (Hb)11.3 g/dL. 

All patients had a rapid virologic decline similar to those with 

normal renal function. Exposure of GS-331007, the primary 

circulating SOF metabolite which is renally eliminated, was 

~6-fold higher and SOF was ~1.4 fold higher than observed 

in subjects in the Phase 2/3 population. Adverse events (AEs) 

reported in more than one patient were anemia (n=3) and 

dizziness (n=2). Two patients had treatment-emergent SAEs, 

1 patient discontinued treatment just before week 12 due to 

an SAE of acute respiratory failure, and 1 patient experienced 

hematemesis. No patients had RBV dose-reduction, interruption 

or discontinuation with the exception of the one patient who 

discontinued study treatment due to an SAE. Three patients 

on epoetin at baseline continued throughout treatment. No 

patients started epoetin or had a blood transfusion during the 

treatment period. Changes in renal function, hemoglobin, and 

cardiac parameters are shown in Table 1. Conclusions: SOF 

400mg daily + RBV 200mg in GT1 or 3 HCV-infected patients 

with severe renal impairment was well-tolerated and resulted in 

rapid virologic suppression through Wk 12 of treatment. Final 

safety, PK and efficacy (SVR12) will be presented.


Table 1. Mean changes in clinical parameters through Week 12 

on treatment 

Disclosures: 






Lin Liu - Employment: Gilead Sciences, Inc. 

Karim Sajwani - Employment: Gilead Sciences, Inc. 

Brian Kirby - Employment: Gilead Sciences 

Jill M. Denning - Employment: Gilead Sciences, Inc. 



Sciences 


Sciences 











The following authors have nothing to disclose: Grisell Ortiz-Lasanta, Richard 

A. Robson 

1129 

A Meta-Analysis of the Association Between Pre-Treatment 

Variables and SVR Amongst Genotype 1 Patients 

Treated with ABT-450/r–Ombitasvir and Dasabuvir 

(VIEKIRA PAK) plus Ribavirin for 12 Weeks 

Gregory J. Botwin 2 , Timothy R. Morgan 1,3 ; 1 Gastroenterology (11), 

VA Long Beach Healthcare System, Long Beach, CA; 2 Research 

Service, VA Long Beach Healthcare System, Long Beach, CA; 

3 Division of Gastroenterology, University of California - Irvine, 

Irvine, CA 

Background: VIEKIRA PAK plus Ribavirin (RBV) is a potent, all 

oral, treatment regimen indicated for the treatment of chronic 

hepatitis C (HCV) genotype (GT)-1. Due to the overall high 

rates of sustained virologic response (SVR) achieved in Phase 

II and Phase III clinical trials, little is known about the association 

between pre-treatment variables and SVR. Purpose: To 

perform a meta-analysis to examine the association between 

pre-treatment variables and SVR in GT-1 patients. Methods: 

We reviewed all registration trials for VIEKIRA PAK. Trials 

that treated GT-1a, or GT-1a and GT-1b, monoinfected, non-cirrhotic, 

HCV patients for 12 weeks with VIEKIRA PAK plus 

RBV were included in our initial analysis. Trials that enrolled 

exclusively GT-1b patients were excluded due to the overall 

high SVR rates. Relative risk (RR) was calculated for the following 

pre-treatment variables: age, body-mass index (BMI), ethnicity, 

IL28B, race, sex, and viral load (VL). A binary random 

effects model was used in the meta-analysis. Results: Three 

studies (SAPPHIRE-I and II, and PEARL-IV) and 870 patients met 

the criteria. Of the evaluated patients 595 were genotype 1a, 

and 297 were treatment experienced. Compared with patients 

with a BMI < 30, a BMI ≥ 30 was associated with a reduced 

RR of SVR, 0.948 (95% confidence-interval [CI], 0.903-0.995, 

p=0.029). The overall SVR in each group was 92% (139/151; 

BMI ≥ 30) and 97% (699/719; BMI < 30). Patients with a VL 

≥ 800,000IU/mL (SVR 96%, 686/716) also had a reduced 

RR of SVR, 0.973 (CI 0.948-0.999, p=0.041) compared to 

patients with a VL below this level (SVR 99%, 152/154). Neither 

age < 55 years, Hispanic ethnicity, IL28B CC genotype, 

African American race, nor male sex were found to be significantly 

positively or negatively associated with SVR. When 

380 GT-1 cirrhotic patients, treated for 12 or 24 weeks (TUR- 

QUOISE-II) were included in the analysis, only BMI remained 

significantly associated with SVR (RR 0.956, CI 0.921-0.993, 

p=0.021). Conclusion: Genotype 1 non-cirrhotic patients with 

an elevated BMI or VL are associated with a reduced chance 

of achieving SVR when treated with VIEKIRA PAK plus RBV 

for 12 weeks. Research evaluating the mechanism for these 

differences may be warranted. 

Pre-Treatment Variables Association with SVR 

Disclosures: 

Timothy R. Morgan - Grant/Research Support: Merck, Abbvie, Genentech, Gilead, 

Bristol Myers Squibb 

The following authors have nothing to disclose: Gregory J. Botwin 

1130 

On-treatment HCV RNA Decline in Pre-and Post-Liver 

Transplant Patients with Different Degrees of Fibrosis 

and Cirrhosis: a combined analysis of the SOLAR trials 

Tania M. Welzel 1 , K. Rajender Reddy 2 , Michael P. Manns 3 , Steven 

L. Flamm 4 , David J. Mutimer 5 , Edward J. Gane 6 , Robert H. 

Hyland 7 , Ming Lin 7 , Phillip S. Pang 7 , John G. McHutchison 7 , Didier 

Samuel 8 , Michael R. Charlton 9 , Xavier Forns 10 , Gregory T. Everson 

11 , Stefan Zeuzem 1 , Nezam H. Afdhal 12 ; 1 Johann Wolfgang 

Goethe University Medical Center, Frankfurt am Main, Germany; 

2 University of Pennsylvania School of Medicine, Philadelphia, PA; 

3 Hannover Medical School, Hannover, Germany; 4 Northwestern 

University Feinberg School of Medicine, Chicago, IL; 5 Queen Elizabeth 

Hospital and University of Birmingham, Birmingham, United 

Kingdom; 6 University of Auckland, Auckland, New Zealand; 7 Gilead 

Sciences, Inc, Foster City, CA; 8 Université Paris-Sud, Villejuif, 

France; 9 Intermountain Medical Center, Murray, UT; 10 Liver 

Unit, IDIBAPS and CIBEREHD, Barcelona, Spain; 11 University of 

Colorado Denver, Aurora, CO; 12 Beth Israel Deaconess Medical 

Center, Boston, MA 

Background and Aims: In the SOLAR-1 and SOLAR-2 studies, 

ledipasvir/sofosbuvir (LDV/SOF)+ribavirin (RBV) for 12 or 24 

weeks resulted in high SVR rates in genotype 1 or 4 HCV-infected 

patients with decompensated cirrhosis or who were liver 

transplant recipients. In this large combined post hoc analysis 

of on-treatment HCV RNA levels in SOLAR-1 and SOLAR-2 study 

participants we investigate whether on-treatment response varied 

by patient population and/or was predictive of treatment 

outcome. Methods: Data from the identically designed SOLAR1 

(NCT01938430) and SOLAR 2 (NCT02010255) studies were 

combined. Six groups of genotype 1 or 4 HCV-infected patients 

were randomized to receive 12 or 24 weeks of LDV/SOF+RBV 

treatment: patients without transplant and either 1) Child-Pugh- 

Turcotte (CPT) B cirrhosis, or 2) CPT C cirrhosis; or patients 

who have undergone transplantation and who were either 3) 

without cirrhosis (F0 to F3), 4) CPT A cirrhosis, 5) CPT B cirrhosis, 

or 6) CPT C cirrhosis. Patients with FCH were excluded


from this analysis. For analysis of early viral kinetics, the 12 

and 24 week treatment durations were combined. Serum HCV 

RNA was quantified using Roche CAP/CTM v2.0 with a lower 

limit of quantitation (LLOQ) of 15 IU/mL. Results: Patients with 

advanced liver disease had SVR12 rates of 86-89%. Patients 

who were post liver transplant had SVR rates that ranged from 

96%-98% in those with F0-F3 fibrosis to 60-75% of patients 

with severe hepatic impairment. Rapid HCV RNA declines 

were observed in all treatment groups. The majority of subjects 

achieved HCV RNA


Disclosures: 

Benjamin Maasoumy - Advisory Committees or Review Panels: Abbott Molecular, 

Janssen-Cilaq; Grant/Research Support: Abbott Molecular; Speaking and Teaching: 

MSD, Roche Diagnostics, Roche Pharma, Janssen-Cilaq, Fujirebio, BMS 



Kerstin Port - Advisory Committees or Review Panels: Janssen; Speaking and 

Teaching: Roche, Gilead, MSD, Janssen 





Novartis 









The following authors have nothing to disclose: Christoph Hoener zu Siederdissen, 

Katja Deterding, David J. Back 

1132 

Interferon Signaling and the Innate Immune Response: 

Still Relevant in the Age of Interferon Free Treatment 

Kate E. Childs, Ivana Carey, William M. Swanson, Suman Verma, 

Kosh Agarwal; Institute of Liver Studies, King’s College Hospital, 

London, United Kingdom 

Chronic innate immune activation predicts non-response to 

interferon based treatment for Hepatitic C Virus (HCV). Predictors 

of response to DAAs are not well characterized. Methods: 

46 patients (26 F, 20 M, median age 58 (51, 63) with HCV 

decompensated liver cirrhosis, Childs Pugh B were treated with 

12 weeks of an IFN-free regimen (77%SOF/ledipasvir+RBV 

and 23%SOF+DCV+RBV). EDTA plasma was taken at baseline 

(TW0), treatment week 4 (TW4), TW12, SVR 4 and SVR 

12. ELISAs for IP-10, CD26 which cleaves IP-10 to a shorter 

antagonistic form, IFN A, G L1, 2 and 3 were performed. 

Results: 37 patients responded (RES) to therapy, achieving SVR 

12. 9 patients experienced HCV relapse (REL) post TW12. At 

baseline, median IP-10 was higher in REL, CD26 trend higher 

(p=0.053) Baseline IFN G, L1, L2 and L3 did not differ. The 

kinetics of IP-10 and CD26 differed significantly between RES 

and REL,(fig) IP-10 was higher in REL and fell at TW4, IP-10 

increased in RES at TW4. Univariate logistic analysis was 

carried out for HCV genotype, biochemical parameters, HCV 

RNA, baseline IP10 and CD26, baseline IFN G and L and 

changes in IP-10 and CD26 on therapy. In the final model only 

baseline IP-10 and change in IP-10 at TW4 were significant 

(p=0.043 and p=0.045 respectively) This model was able to 

predict the treatment outcome in 95% of cases. Discussion- In 

this group of patients with decompensated cirrhosis, IP-10, a 

surrogate marker of hepatic ISG expression and its changes 

early in treatment predicted response to DAA treatment. REL 

had high baseline IP-10 which then decreased on treatment, 

whereas RES had lower baseline levels and an increase on 

treatment, which may mirror findings from the peg-interferon 

era where high baseline ISG expression could not upregulate 

in response to exogenous interferon. More work is warranted 

to investigate predictors of treatment outcome. These data suggest 

that the innate interferon driven response to HCV remains 

relevant as a predictor of response to interferon free regimens 

for HCV. 

Disclosures: 


BMS 




The following authors have nothing to disclose: Kate E. Childs, William M. 

Swanson, Suman Verma 

1133 

Pharmacokinetic Analyses of Ledipasvir/Sofosbuvir and 

HIV Antiviral Regimens in Subjects with HCV/HIV Co-infection. 

Polina German, Liyun Ni, Luisa M. Stamm, Jenny C. Yang, Maryanne 

Lenoci, John Ling, Anita Mathias; GIlead Sciences, Inc, Foster 

City, CA, CA 

Background Administration of ledipasvir (LDV)/sofosbuvir 

(SOF) fixed-dose combination tablet once-daily for 12 weeks 

achieved overall SVR12 rates of 96% (322/335) in treatment-naïve 

and treatment-experienced HCV/HIV co-infected 

subjects (ION4). Pharmacokinetic (PK) data were collected 

to examine the relationship between exposure and extrinsic/ 

intrinsic variables or treatment outcome and to compare PK in 

co-infection and HCV mono-infection. Methods Treatment-naïve 

(N=150) and treatment-experienced (N=185) subjects on a 

stable ARV regimen of efavirenz (EFV 600 mg QD), rilpivirine 

(RPV; 25 mg QD) or raltegravir (RAL; 400 mg BID or 800 

mg QD) with a backbone of emtricitabine/tenofovir DF (FTC/ 

TDF; 200 mg/300 mg) received LDV/SOF 90mg/400mg for 

12 weeks. The PK of LDV, SOF, GS-331007 (SOF predominant 

circulating metabolite) and tenofovir (TFV) were evaluated 

in all subjects with measurable plasma concentrations using 

previously established population PK models. EFV, RPV, RAL 

and FTC PK were evaluated using noncompartmental methods 

in subjects with intensive PK sampling data. LDV, SOF and 

GS-331007 exposures in HCV/HIV co-infected subjects were 

compared across ARV regimens, race and treatment outcome, 

and to subjects with HCV monoinfection. The PK of ARVs was 

descriptively compared to historical data from HIV monoinfection 

studies. Results Exposures of LDV, SOF and GS-331007 

were comparable across ARV regimens, race and treatment 

outcome. There were no clinically relevant differences in the 

PK of LDV/SOF in HCV/HIV co-infected subjects compared to 

HCV-monoinfected subjects. EFV, RPV, RAL and FTC PK were 

consistent with historical data. In agreement with the findings 

from the Phase 1 evaluations, which revealed higher TFV 

exposure with LDV/SOF, TFV PK was modestly higher than 

observed historically with NNRTI or InSTI-based regimens plus 

FTC/TDF. Conclusion PK results in conjunction with safety and 

efficacy data support the use of LDV/SOF 90 mg/400 mg for


12 weeks in HCV/HIV co-infection. Higher TFV exposures are 

observed in the presence of LDV/SOF; therefore, monitoring 

for TFV-associated AEs is warranted during co-administration 

of LDV/SOF and TDF-based regimens. 

Disclosures: 

Polina German - Employment: GIlead Sciences, Inc; Stock Shareholder: GIlead 

Sciences, Inc 



Anita Mathias - Employment: Gilead Sciences Inc., 

The following authors have nothing to disclose: Liyun Ni, Maryanne Lenoci, John 

Ling 

1134 

HCV genotype 3: Meta-analysis with current options 

Javier Ampuero 1 , K. Rajender Reddy 2 , Manuel Romero-Gomez 1 ; 

1 Unit for the Clinical Management of Digestive Diseases, Valme 

University Hospital, Sevilla, Spain; 2 Division of Gastroenterology 

and Hepatology, Hospital of the University of Pennsylvania, 

Philadephia, PA 

Background & Aim: Landscape of HCV therapy is changing 

rapidly with new direct acting antivirals (DAAs), with sustained 

virological response (SVR) rates around 90%. However, genotype 

3 represents the new challenge, as these patients show 

lower SVR rates, particularly cirrhotic and treatment-experienced 

patients. Many treatment options have been used but the 

definite therapy remains unclear. Our aim was to address therapeutic 

efficacy of the various treatment regimens through a 

meta-analysis in HCV genotype 3: a) sofosbuvir (SOF) + peginterferon 

(IFN) + ribavirin (RBV) 12 weeks vs. SOF+RBV 24w 

(n=645); b) SOF+RBV 12w/16w vs. SOF+RBV 24w (n=503); 

c) SOF + daclatasvir (DCV) vs. SOF+DCV+RBV (n=162); d) 

SOF + ledipasvir (LDV) vs. SOF+LDV+RBV (n=169). Methods: 

We reviewed and identified studies in Pubmed and EMBASE 

until June 2015. We included certain number of published 

papers and presented abstracts (last AASLD and EASL meetings) 

in genotype 3 patients randomized to at least, 2 arms 

of HCV therapy. The random effect models of Der Simonian 

and Laird method were used for heterogeneous studies using 

the Meta-Disc software 1.4 (Madrid, Spain). Results: Patients 

taking SOF+INF+RBV 12w (92.9%; 196/211) showed higher 

SVR rates than those treated with SOF+RBV 24w (79.4%; 

201/253) (OR 2.67 (95%CI 1.43-4.98); I 2 =0%) (Figure). 

Twenty-four weeks of combination SOF+RBV had higher SVR 

rates (84.4%; 391/463) than those treated for 12/16w 

(70.7%; 157/222) (OR 3.45 (95%CI 1.02-11.11); I 2 =70%). 

In SOF+DCV-based therapies, the addition of RBV (90.3%; 

28/31) did not enhance SVR rates significantly (81.7%; 

107/131) (OR 1.16 (95%CI 0.44-3.08); I 2 =0%). In contrast, 

the addition of RBV ((81%; 111/137) vs. (62.5%; 20/32)) was 

able to increase the SVR rates in patients treated with SOF+LDV 

(OR 3.30 (95%CI 1.35-8.33); I 2 =0%). Conclusion: Triple therapy 

of SOF+IFN+RBV has demonstrated better SVR rates than 

IFN-free regimens in genotype 3 patients. To optimize SVR, 

interferon free regimen of SOF+RBV needs to be administrated 

over longer period (24 weeks) rather than shorter duration of 

12w-16 weeks. With SOF+LDV combination, RBV is essential 

to increase SVR rates probably because LDV has a very low 

activity against genotype 3. By contrast, the addition of RBV in 

DCV-based treatment does not increase SVR. 

Disclosures: 






S.A. 

The following authors have nothing to disclose: Javier Ampuero 

1135 

Utilization and Effectiveness of Sofosbuvir Based HCV 

Regimens in a Community Based Setting 

T. Craig Cheetham 1 , Sara Y. Tartof 1 , Zoe Bider-Canfield 1 , Vincent 

Louie 3 , Derenik Gharibian 3 , Amandeep K. Sahota 2 ; 1 Research & 

Evaluations Department, Kaiser Permanente Southern California, 

Pasadena, CA; 2 Gastroenterology/Hepatology, Southern California 

Permenente Medical Group, Los Angeles, CA; 3 Pharmacy 

Department, Kaiser Foundation Hospitals, Los Angeles, CA 

Background: High rates of sustained virologic response have 

been reported in the clinical trials for sofosbuvir (Sof) based 

regimens. Less information is available regarding the effectiveness 

(sustained virologic response 12 weeks, SVR12) of Sofbased 

regimens outside of clinical trials. Objective: Determine 

the rates of SVR12, for Sof-based regimes, in a community 

setting. Methods: Kaiser Permanente Southern California is an 

integrated health-care delivery system providing medical services 

to 3.8 million members. Adults started on a Sof-based 

regimen beginning 12/1/2013 and completing treatment by 

1/31/2105 were eligible. Six months continuous membership 

prior to starting therapy was required. Co-administered 

drugs included pegylated-interferon (PEG), simeprevir (Sim) 

and ribavirin (Riba). Baseline co-morbid conditions, viral load, 

genotype (GT), and prior Hepatitis C (HCV) treatments were 

collected from the electronic medical record. Viral load tests 

were collected through 4/30/2015 allowing 12 weeks follow-up 

for all patients. Descriptive and multivariable analyses 

were conducted to evaluate the utilization and outcomes of 

Sof-based regimens. SVR12 was the primary endpoint. Results: 

A total of 355 patients were identified. Twelve patients did not 

complete therapy and were excluded (2 died, 6 experienced 

adverse events, and 4 were stopped for non-adherence). For 

the cohort the average age was 58.5 years and 64.7% were 

male. Cirrhosis was present at baseline in 39.7% of patients 

and 43.7% were treatment experienced. Genotypes 1, 2 & 3 

accounted for 97.1% of the HCV cases (100 - GT1, 194 - GT2, 

39 - GT3). The most common regimens were Sof-Rib (n=254), 

Sim-Sof (n=12) and Sof-PEG-Rib (n=75). The overall SVR12 

rate was 82.2%. The table has a breakdown of SVR12 by 

genotype and major risk factors. In general, higher rates of 

SVR12 were seen in treatment naïve and non-cirrhotic patients. 

Rates of SVR12 were greater than 90% in GT2 patients (almost 

all received Sof-Riba). In the multivariable logistic regression 

models, cirrhosis was associated with a higher risk of failure. 

Conclusion: In a community based setting, Sof base regimens 

were associated with high SVR12 rates in GT2 patients. Significantly 

lower rates of SVR12 were seen in GT1 and GT3, which 

may be due to the high number of treatment experienced and 

cirrhotic patients.


SVR12 Rates by Major Risk Factors 

Demographics(%) 

Disclosures: 


The following authors have nothing to disclose: Sara Y. Tartof, Zoe Bider-Canfield, 

Vincent Louie, Derenik Gharibian, Amandeep K. Sahota 

1136 

Evaluation of an Integrated HIV/HCV Care Model in an 

Urban, Low-Income, HIV/HCV Co-Infection Clinic 

Angela Kapalko 1 , Linden Lalley-Chareczko 1 , Meghan F. Fibbi 2,3 , 

Karam Mounzer 1,4 ; 1 Jonathan Lax Treatment Center, Philadelphia 

FIGHT, Philadelphia, PA; 2 Department of Family and Community 

Medicine, University of Pennsylvania, Philadelphia, PA; 3 Philadelphia 

College of Osteopathic Medicine, Philadelphia, PA; 4 Perelman 

School of Medicine, University of Pennsylvania, Philadelphia, 

PA 

Background The Jonathan Lax Treatment Center is an urban, 

low-income, Ryan White and FQHC funded, outpatient medical 

center providing services via an integrated care model to 

approximately1700 patients with HIV, of whom 459 are HIV/ 

Hepatitis C(HCV) co-infected. The HIV/HCV co-infection treatment 

protocol at the Lax Center incorporates on-site disease 

staging, pharmacy refills/pill-packing services with flexible 

dispensation schedules, adherence monitoring, case management 

and mental health services.The purpose of the present 

observational study is to evaluate the comprehensive HIV/HCV 

co-infection treatment protocol in a patient-centered care setting 

at the Lax Center. Methodology A comprehensive chart review 

of all Lax Center patients carrying a diagnosis of chronic 

HCV between the calendar years of 2011 and 2014(n=459) 

was performed. Patient demographics, HCV genotyping and 

disease staging, prior HCV treatment experience, HCV sustained 

virologic response(SVR) and information on comorbid 

conditions present during HCV treatment were recorded. All 

co-infected patients were included in the overall demographic 

analysis; however, patients treated on a clinical trial were 

excluded from the treatment outcome analyses. Results 459 

co-infected patient records active between Jan 1, 2011 and 

Dec 31 2014 were reviewed. 88 individual patients(19.1%, 

n=459) received treatment during this time frame, 10 of whom 

were retreated, making the total number of treatments delivered 

98. 26 treatments were excluded due to participation in 

clinical trials, leaving 72 eligible treatments for final analyses. 

The demographics of those treated were similar to the overall 

Lax HIV/HCV population (See Table 1). 31.9% of all treatment 

efforts consisted of Telaprevir based regimens with an 

SVR12 rate 56.5%(n=23). IFN free regimens (SOF/RBV, SOF/ 

SMV, SOF/LDV)(n=43) yielded an SVR12(n=39) of 87.1% 

with 4 SVR12 results pending(2 confirmed SVR4). No patients 

were lost-to-follow-up while on treatment or during post-treatment 

monitoring. Conclusions Operating under a patient-centered, 

integrated care model, the Lax Center was able to treat 

19.1%(n=88) of active HIV/HCV patients, retain 100% in 

care, and achieved an overall SVR rate of 75.0%(with 4 SVR 

12 pending). These data support the utility of an integrated 

care model for optimization of treatment outcome among HIV/ 

HCV co-infected patients in an urban, low-income community 

setting. 

Disclosures: 

Angela Kapalko - Advisory Committees or Review Panels: Gilead Sciences, Abbvie; 

Speaking and Teaching: Abbvie 

Karam Mounzer - Advisory Committees or Review Panels: Gilead Sciences, BMS, 

J& J, Merck; Grant/Research Support: Boehrlingher Ingelheim, Gilead Sciences, 

Glaxo Smith Kline ViiV, Janssen and Janssen, Merck; Speaking and Teaching: 

Gilead Sciences, BMS, J& J, Merck 

The following authors have nothing to disclose: Linden Lalley-Chareczko, Meghan 

F. Fibbi 

1137 

What does Fibroscan® measure shortly after IFN-free 

DAA therapy? 

Karin Kozbial, Albert Stättermayer, Clarissa Freissmuth, Rafael 

Stern, Sandra Beinhardt, Christian Rechling, Petra E. Steindl- 

Munda, Michael Trauner, Peter Ferenci, Harald Hofer; Internal 

Medicine 3, Medical University of Vienna, Vienna, Austria 

Background:Sustained virological response (SVR) by any antiviral 

therapy may improve liver fibrosis in the majority of patients 

during long-term follow up. Interferon free combinations of 

direct acting antivirals (DAAs) achieve a rapid and profound 

inhibition of HCV replication during treatment and offer the 

possibility to study short-term changes in fibrosis.Therefore 

this study investigated liver stiffness in patients with advanced 

fibrosis (F3-F4) shortly after DAA treatment. Methods:Liver stiffness 

was measured before and at the end or up to 12 weeks 

after the end of DAA-therapy (Sofosbuvir[SOF] combined with 

daclatasvir[DCV, n=52], ribavirin [n=10], ledipasvir [n=14], 

Simeprevir [SMV, n=24], or the combination of DCV and SMV 

[n=9])by Fibroscan®(FS; Echosens 502, Paris, France) in 109 

patients (f/m=40/69; mean age: 55.2±10.5yrs; genotype 

(GT) 1: 87, GT3: 16, GT4: 6; F3/F4=28/81; BMI 27.1±4.4). 

Treatment duration was 12 (n=48) up to 24 weeks (n=61). 

Only FS results with IQR ≤ 30% were included. Liver fibrosis categories 

were defined as advanced fibrosis (F3:9.6-12.4kPa), 

and cirrhosis (F4: ≥12.5kPa). Results:Overall, liver stiffness 

decreased from 23.2kPa (95% CI 20.4-26.0) to 20.1kPa (95% 

CI 17.0-23.3;p


Harald Hofer - Advisory Committees or Review Panels: Gilead, Abbvie; Speaking 

and Teaching: Janssen, BMS, Gilead, Abbvie 

The following authors have nothing to disclose: Karin Kozbial, Albert Stättermayer, 

Clarissa Freissmuth, Rafael Stern, Sandra Beinhardt, Christian Rechling, 

Petra E. Steindl-Munda 

1138 

Association between severe rash and HLA polymorphism 

in telaprevir-based triple therapy for chronic hepatitis 

C in a multi-centric study in Japan 

Yoko Yamagiwa 1 , Naohiko Masaki 1 , Nao Nishida 1 , Minae 

Kawashima 2 , Seik-Soon Khor 2 , Hiroko Miyadera 1 , Masaya Sugiyama 

1 , Masaaki Korenaga 1 , Kazumoto Murata 1 , Tatsuya Kanto 1 , 

Yasuhito Tanaka 3 , Kazuhiko Koike 4 , Katsushi Tokunaga 2 , Masashi 

Mizokami 1 ; 1 The Research Center for Hepatitis and Immunology, 

National Center for Global Health and Medicine, Ichikawa, Japan; 

2 Department of Human Genetics, Graduate School of Medicine 

The University of Tokyo, Tokyo, Japan; 3 Department of Virology 

and Liver Unit, Nagoya City University Graduate School of Medical 

Sciences, Nagoya, Japan; 4 Department of Gastroenterology, 

The University of Tokyo, Tokyo, Japan 

Background and Aim Although telaprevir (TVR) enabled a 

remarkable improvement in efficacy of interferon treatment 

for chronic hepatitis C, it had serious adverse events such as 

dermatological adverse events which required discontinuance 

of treatment in serious grade of rash which was potentially 

life-threatening. However, mechanism of severe rash in TVRbased 

therapy has not been clarified yet. Thus, we aimed to 

identify genetic risk factors associated with serious grade of 

rash. Methods and Results Genome-wide association study 

(GWAS) was performed in 384 patients received TVR combined 

with ribavirin and peg-interferon using Affymetrix 

AXIOM ASI arrays. We compared allele frequency of each single 

nucleotide polymorphism (SNP) between 182 patients who 

did not experience skin eruption and 50 patients with grade 3 

of rash defined by rash over 50% of body surface area, rash 

with additional changes such as vesicles, or with systematic 

reaction such as fever. However, no significant association 

was identified in the GWAS stage. Next, SNP imputation and 

association testing was performed using PLINK in 141 SNPs 

with p < 1.00E-05 in the GWAS stage. As a result, 22 out 

of 25 SNPs with p < 1.00E-06 in the SNP imputation, were 

located in the human leukocyte antigen (HLA) region on chromosome 

6. Finally, statistical imputation analysis of classical 

HLA alleles was performed in the SNP data using HIBAG R 

package optimized for Japanese population (Pharmacogenomics 

J. 2015). HLA-DRB1*09:01 showed a significant association 

with an odds ratio (OR) of 3.376 (p = 9.12E-06) and 

HLA-DQB1*03:03, which is known to be in strong linkage 

disequilibrium with DRB1*09:01 in the Japanese population, 

also showed a significant association with an OR of 2.907 (p 

= 7.15E-05). Discussion and Conclusions More frequent severe 

rash in phase III trial in Japan (9%) compared with that in the 

USA and Europe (5%), may related with higher frequency of 

HLA-DRB1*09:01 in Japanese population than in Caucasian 

(15% vs 1%). Genetic variants located in the HLA region in 

Japanese should be candidate genetic factors in susceptibility 

to severe rash in TVR-based triple therapy. 

Disclosures: 



Squibb 

The following authors have nothing to disclose: Yoko Yamagiwa, Naohiko 

Masaki, Nao Nishida, Minae Kawashima, Seik-Soon Khor, Hiroko Miyadera, 

Masaya Sugiyama, Masaaki Korenaga, Kazumoto Murata, Tatsuya Kanto, 

Kazuhiko Koike, Katsushi Tokunaga, Masashi Mizokami 

1139 

Patient Reasons for Initiating or Delaying Hepatitis C 

Virus (HCV) Treatment and Physician Reasons for Selecting 

an HCV Treatment: A Prospective Observational 

Study 

Shelagh M. Szabo 2 , David R. Walker 1 , Timothy R. Juday 1 , 

Suzanne Lane 2 , Sammy Saab 3 ; 1 HEOR, Abbvie, North Chicago, 

IL; 2 Icon Health Economics, Vancouver, BC, Canada; 3 David Geffen 

School of Medicine, UCLA, Los Angeles, CA 

BACKGROUND This is a prospective, multicenter observational 

study of humanistic and economic outcomes among patients 

initiating direct acting antiviral treatments for hepatitis C virus 

(HCV). The objective of this analysis is to describe patient reasons 

for treatment delay or initiation and physician reasons 

for selection of a specific treatment regimen. METHODS All 

consecutive HCV patients attending clinics at ten sites in the 

United States were screened to identify the cohort initiating 

treatment for HCV. From that cohort, data were collected using 

patient-completed surveys at up to five time points over the treatment 

course, supplemented by a medical chart review at treatment 

initiation and completion. Type of data collected included 

demographic, clinical, economic, and humanistic, however, for 

this analysis, only screening data were used. Data on patients’ 

reasons for initiating or delaying treatment, and physicians’ 

reasons for recommending treatment were collected; multiple 

responses were allowed per patient. The first patient enrolled 

January 2013 and the last patient enrolled September 2014. 

RESULTS There were 143 patients, from eight clinics, enrolled 

in the study out of the 787 that were screened across ten clinics. 

Among enrolled patients, 137 were from community clinics 

and 6 were from academic/hospitals. The mean age was 56 

years, 52% male, 60% white, 20% Hispanic and 15% black. 

75% graduated from high school and 33% were employed. 

100 patients were on a sofosbuvir and/or simeprevir-based 

regimen, and 43 patients were on other treatments. 50% were 

on an interferon-free regimen. Patient-reported reasons for initiating 

treatment were desire to be cured (74.1%), physician’s 

recommendation (70.6%), to prevent future liver damage 

(62.2%), peace of mind (49%), to manage HCV symptoms 

(31.5%), worry about transmission (26.6%), to avoid social 

stigma (10.5%), desire to have children (7.7%), family pressure 

(6.3%), and other (2.8%). Patient-reported reasons for initially 

delaying treatment were physician recommendation (23.8%), 

waiting for the availability of new treatments (21.0%), no or 

mild symptoms (17.5%), cost (9.1%), health conditions other 

than HCV (5.6%) and other (39.2%). The primary physician-reported 

reason for selection of a specific treatment regimen was 

likelihood of treatment success (83.9%), followed by tolerability 

(7.7%), reimbursement/access (2.8%), co-pay (0%), and other 

(4.9%). CONCLUSIONS The potential for achieving cure was 

the most important reason for a physician choosing a specific 

HCV treatment regimen. Cost however, did not play an important 

role for treatment selection or delay for physicians and 

patients, respectively. 

Disclosures: 

Shelagh M. Szabo - Employment: Oxford Outcomes 


Healthcare 






The following authors have nothing to disclose: Suzanne Lane


1140 

Co-morbidities and co-medications of patients with 

chronic hepatitis C (CHC) under specialist care in the UK 

- Challenges for scaling up HCV treatment? 

Benjamin E. Hudson 3,8 , William Irving 4,5 , Stephen T. Barclay 1 , 

Fiona Marra 2,6 , Alex J. Walker 7 ; 1 Walton Liver Clinic, Glasgow, 

United Kingdom; 2 NHS Greater Glasgow and Clyde, Glasgow, 

United Kingdom; 3 HCV Research UK, Nottingham, United Kingdom; 

4 NIHR Nottingham Digestive Diseases Biomedical Research 

Unit, Nottingham, United Kingdom; 5 School of Life Sciences, University 

of Nottingham, Nottingham, United Kingdom; 6 University of 

Liverpool, Liverpool, United Kingdom; 7 University of Nottingham, 

School of Life Sciences, Nottingham, United Kingdom; 8 Cambridge 

University Hospitals NHS trust, Cambridge, United Kingdom 

Introduction Most patients with CHC can benefit from direct 

acting antiviral (DAA). Alongside specific CHC disease indicators, 

physical/psychiatric comorbidities, lifestyle factors 

and related co-medications with drug-drug interaction (DDI) 

potential will inform appropriate CHC treatment selection. We 

describe demographics, comorbidities and common co-medications 

of CHC patients currently under specialist care in the 

UK. Methods Retrospective analysis of routinely collected data 

of CHC patients from 59 UK specialist centres enrolled with 

the National HCV Research UK Biobank between 03/2012- 

10/2014. Results 6,278 patients were analysed. Median age 

52 years (IQR 43-59), 70.4% male, 84.7% white. Genotype 

1: 50%, 2: 4.0%, 3: 33.7%, 4: 3.6%, 5/6: 0.3%. Cirrhotic: 

23.6% (age >60’s 36.6%). Acquisition mode: injection drug 

use (IDU) 59.2%, blood products 11.3%, non-UK born 9.4%, 

other 20.1%. Subgroup analysis for IDU acquisition and age 

was performed. Social history (Hx): previous/current heavy 

alcohol use: overall 38.3% (IDU 50.1%), current cannabis use: 

24.6% (IDU 35.2%). Comorbidities: Hx of depression: 45.4% 

(IDU 57.6%), Hx of attempted suicide or inpatient treatment 

for depression: 19.4% (IDU 27.3%). HIV co-infection 5.0% 

(IDU: 4.5%). Prevalence of diabetes, cancer and renal dialysis 

correlated with age (Diabetes: overall 11.3%, >60s 22.9%, 

Cancer: 8.1% vs 18.1%, Dialysis: 1.3% vs 2.2%). Most common 

co-medications with DDI potential were psychotropic with 

38.6% (IDU 53.3%) on either antidepressant (22.9%), opioid 

replacement (21.2%) or hypnotic (10.4%) (includes polypharmacy). 

Prescriptions of antidiabetics, statins and immunosuppressants 

rose sharply with age (60’s 31.7%). 

Conclusions We found high levels of co-morbidity, ongoing 

substance abuse and co-medication with DDI potential in 

CHC populations under specialist care in the UK. Age-related 

increasing levels of advanced liver disease, comorbidities and 

polypharmacy may further increase the challenges in managing 

this patient group. CHC treaters need to be aware of DDI 

potential when choosing appropriate CHC treatments. 

*antidepressants, opioid replacement, hypnotics;** Antiretrovirals;**not 

specified 

Disclosures: 

William Irving - Advisory Committees or Review Panels: Novartis, MSD, Janssen 

Cilag, Bristol Myers Squibb; Grant/Research Support: GSK, Pfizer, Janssen 

Cilag, Gilead Sciences; Speaking and Teaching: Janssen Cilag, Roche 

Stephen T. Barclay - Advisory Committees or Review Panels: Gilead, Janssen, 

MSD, Abbvie, BMS; Speaking and Teaching: Gilead, Janssen 



The following authors have nothing to disclose: Benjamin E. Hudson, Alex J. 

Walker 

1141 

Patterns of Care Since the Approval of New Therapies 

for Hepatitis C Virus (HCV) Infection: A 2013-2014 Real- 

World Analysis of US Community Specialty Practices 

Paul J. Gaglio 2 , Tamar Sapir 1 , Jeffrey D. Carter 1 , Laurence 

Greene 1 , Erica Rusie 1 , Kathleen Moreo 1 ; 1 PRIME Education, Inc., 

Tamarac, FL; 2 Hepatology, Montefiore Medical Center, Bronx, NY 

Background: From 2013 to 2014, approval of all-oral therapies 

was associated with dramatic changes in managing HCV. 

To assess “real-world” use of HCV therapy and management 

trends, we conducted a retrospective study of 50 US community 

practices. Methods: The cohort comprised 17 hepatologists, 

17 gastroenterologists, and 16 infectious disease specialists 

across the US. Charts of adult HCV patients were reviewed for 

2013 (n = 300) and 2014 (n = 500). Charts were abstracted 

for patient and disease characteristics, care processes, adherence 

to HCV quality measures, and prescribing patterns. T-tests 

or chi-square tests were used to analyze differences in means 

for 2013 and 2014. Results: From 2013 to 2014, patient 

demographics, risk factors, disease characteristics, and comorbidities 

were as follows: average age (51 vs 54 yrs p=.07), 

time since diagnosis (8 vs 5 yrs, p=.04), GT1 (75% vs 76%, 

p=.75), GT2 (9% vs 10%, p=.77), GT3 (13% vs 10%, p=.15), 

substance abuse (41% vs 33%, p


1142 

Treatment Outcomes of Veterans with Decompensated 

Cirrhosis Receiving All Oral Direct Acting Antiviral Hepatitis 

C Therapy 

Abigail Rabatin 1 , Mohamed Hashem 1 , Mary L. Townsend 1 , William 

E. Bryan 1 , Cynthia A. Moylan 1,2 , Steve S. Choi 1,2 , Susanna 

Naggie 1,2 ; 1 Durham VA Medical Center, Raleigh, NC; 2 Duke University 

Medical Center, Durham, NC 

Purpose Patients infected with hepatitis C virus (HCV) and decompensated 

cirrhosis are a therapeutic challenge. The purpose of 

this study was to evaluate the effectiveness and safety of oral 

sofosbuvir-based regimens in Veterans with decompensated cirrhosis. 

Design This is a multicenter, retrospective, cohort study 

utilizing the national Veterans Affairs (VA) electronic medical 

record. Veterans with decompensated cirrhosis who received 

a sofosbuvir-based HCV regimen at a VA between 12/06/13 

and 12/31/14 were included. Veterans who received interferon-containing 

regimens were excluded. Possible decompensation 

was identified using VA outpatient prescription data for 

either lactulose, rifaximin or sorafenib. Manual chart review 

was conducted confirming decompensating event (bleeding 

esophageal varices, hepatic encephalopathy, ascites, spontaneous 

bacterial peritonitis or hepatocellular carcinoma). The 

primary objective was sustained virologic response defined as 

an undetectable HCV RNA 12 weeks post-treatment (SVR12). 

This study was approved by the Durham VA IRB. Results A total 

of 7,622 Veterans received sofosbuvir during the study period 

and 517 met criteria based on prescription data for possible 

decompensation. A convenience sample of 250 was assessed 

by manual chart review, of which 150 met inclusion criteria. 

84% (N=126) completed treatment and 16% (N=24) discontinued 

treatment early. The majority of Veterans were male 

(97%), Caucasian (70%), with genotype 1-infection (84%) and 

a mean age of 61 years. Median platelet count was 78 and 

median albumin 3.1. There were 5 patients co-infected with 

HBV and 5 with HIV. SVR12 is provided in Table 1. Relapse 

was the only reason for virologic failure in Veterans completing 

therapy. A majority of Veterans (69%) experienced at least one 

adverse event (AE), most commonly fatigue (26%) and nausea 

(11%). Severe AEs were uncommon (8%, N=12) and included 

3 deaths that were not treatment related. Four patients died 

after end of treatment. Conclusion All oral direct acting antiviral 

regimens containing sofosbuvir are safe in Veterans with 

decompensated cirrhosis; however, relapse rates were high 

in this cohort. Patients with decompensated cirrhosis remain a 

special population with unmet medical need and a high mortality 

regardless of therapy. 

Table 1 

1143 

Cost-effectiveness of ombitasvir/paritaprevir/ritonavir, 

dasabuvir +/- ribavirin (3D±R) in patients with genotype 

(GT) 1 chronic hepatitis C virus (HCV) and human 

immunodeficiency virus (HIV) coinfection compared with 

other standards of care in the US 

Suchin Virabhak 2 , Scott J. Johnson 2 , Hélène Parisé 2 , Timothy R. 

Juday 1 , Alice Wang 1 , Yuri Sanchez 1 , Sammy Saab 3 ; 1 HEOR, 

AbbVie, Mettawa, ID; 2 Medicus Economics Inc., Boston, MA; 

3 Pfleger Liver Institute, UCLA, Los Angeles, CA 

BACKGROUND 3D±R has demonstrated safety and efficacy 

in patients with GT1 HCV and HIV coinfection. Its cost-effectiveness 

has not been evaluated vs no treatment (NT) or other 

standards of care in the US, including sofosbuvir plus ledipasvir 

(SOF+LDV), sofosbuvir plus pegylated-interferon and ribavirin 

(SOF+PR), and SOF+R. METHODS A cost-effectiveness Markov 

model was developed based on a natural HCV history with 13 

health states: 8 disease progression states (F0-F4, decompensated 

cirrhosis, hepatocellular carcinoma, and liver transplant), 

3 sustained virologic response states, a spontaneous clearance 

state, and a mortality state. Transition rates were obtained 

from published literature. Adverse event rates, treatment-related 

disutilities, treatment durations and efficacy rates were based 

on the following clinical trials: TURQUOISE I (3D±R), ERADI- 

CATE (SOF+LDV), PHOTON-1 and PHOTON-2 (SOF+R), and 

P7977-1910 (SOF+PR). Baseline patient characteristics were 

based on TURQUOISE I. Direct medical costs, including HIV 

antiretroviral therapy, were extracted from a systematic literature 

review and drug costs were based on the December 2014 

Red Book. The model had a lifetime horizon with an annual 

discount rate of 3%. Outcomes were measured in quality-adjusted 

life-years (QALYs), lifetime costs, and incremental cost 

effectiveness ratios (ICERs) in the overall coinfected population 

and the treatment-naïve non-cirrhotic subpopulation, where 

data were available. Probabilistic sensitivity analyses (PSA) 

were conducted by varying all parameters simultaneously. 

RESULTS In the overall coinfected population, 3D±R was “dominant” 

over SOF+R (i.e., was less costly and more effective) 

and was cost-effective at a threshold of $50,000 per QALY 

gained compared to NT and SOF+PR. In naïve non-cirrhotic 

coinfected patients, 3D±R was cost-effective compared to NT 

at a $50,000 threshold per QALY gained. Compared with 

SOF+LDV, 3D±R offers 0.1 less QALYs and $11,895 lower 

lifetime costs. 3D±R was the preferred regimen in the majority 

of PSA simulations when QALYs were valued at $50,000 or 

$100,000 each. CONCLUSIONS 3D±R is a cost-effective treatment 

option for GT1 HCV and HIV coinfected patients, both in 

the overall and naïve non-cirrhotic populations, compared to 

NT and other standards of care in the US. 

Disclosures: 

Susanna Naggie - Advisory Committees or Review Panels: BMS, Gilead, Abb- 

Vie, Merck; Grant/Research Support: Gilead, AbbVie, BMS, Jenssen, Merck, 

Achillion 

The following authors have nothing to disclose: Abigail Rabatin, Mohamed 

Hashem, Mary L. Townsend, William E. Bryan, Cynthia A. Moylan, Steve S. Choi 

NA: No clinical trial data available 

Disclosures: 

Suchin Virabhak - Consulting: AbbVie 

Scott J. Johnson - Consulting: AbbVie; Employment: Medicus Economics 

Hélène Parisé - Consulting: MedicusEconomics LLC, AbbVie ; Employment: Statlog 

Consulting Inc 


Yuri Sanchez - Employment: AbbVie; Stock Shareholder: AbbVie







1144 

Lifetime risks of liver morbidity and mortality in patients 

with recurrent genotype 1 hepatitis C virus infection 

post-liver transplantation treated with ombitasvir/paritaprevir/ritonavir, 

dasabuvir and ribavirin (3D+R) vs 

former standards of care 

Sammy Saab 2 , Caroline Huber 3 , Alice Wang 1 , Yuri Sanchez 1 , 

Timothy R. Juday 1 ; 1 HEOR, AbbVie, Mettawa, ID; 2 Pfleger Liver 

Institute, UCLA, Los Angeles, CA; 3 Precision Health Economics, Los 

Angeles, CA 

PURPOSE Liver transplant recipients with hepatitis C virus 

(HCV) infection historically have had limited treatment options. 

Without effective treatment, HCV can cause progressive liver 

disease in a short period of time. 3D+R is a recently approved 

interferon-free regimen with demonstrated safety and efficacy 

in the post-transplant genotype (GT) 1 HCV population and its 

long-term impact on liver outcomes is not yet fully understood. 

We assess the lifetime risks of liver morbidity and mortality 

for post-liver transplant patients with GT1 HCV recurrence 

receiving no treatment (NT) or treatment regimens with FDA 

approval or Phase III trial results. METHODS A US-based, twophase 

Markov health state model estimated outcomes over a 

lifetime horizon for a cohort of post-liver transplant patients 

with recurrent GT1 HCV and METAVIR fibrosis stage F0-F2. 

In the first phase, patients were distributed into disease states, 

including sustained virologic response, based on the efficacy of 

three treatment options: NT, pegylated interferon and ribavirin 

for 48 weeks (PR48), or 3D+R for 24 weeks. The second phase 

followed patients’ progression through compensated cirrhosis 

(CC), decompensated cirrhosis (DCC) and death. Transition 

rates were extracted from published literature. Population characteristics 

and treatment efficacy rates for PR48 and 3D+R 

were based on Phase III clinical trials in post-liver transplant 

patients. Lifetime risks of CC, DCC and 10-year all-cause mortality 

were analyzed. RESULTS The lifetime risks of liver morbidity 

and all-cause mortality were substantially lower for GT1 

HCV patients with liver transplant treated with 3D+R than those 

receiving NT or PR48. For a representative patient, the lifetime 

risk of developing CC was 63.3% with NT, 51.5% with PR48, 

and 1.9% with 3D+R. The lifetime risk of developing DCC 

was 55.6% with NT, 45.3% with PR48 and 1.7% with 3D+R. 

All-cause mortality after 10 years was 29.5% with NT, versus 

27.4% with PR48 and 18.6% with 3D+R treatment. CONCLU- 

SIONS The likelihood of liver transplant recipients infected 

with GT1 HCV experiencing liver morbidity (i.e., CC or DCC) 

or all-cause mortality 10 years post-treatment was substantially 

reduced with 3D+R compared to patients treated with PR48 or 

not treated. 

Disclosures: 





Caroline Huber - Employment: Precision Health Economics 




1145 

Quantifying and Predicting Early Clinical Gains with 

Direct Acting Antiviral Therapy in Well Compensated 

HCV Cirrhosis: Is There an Unrecognized “Point of 

No MELD-based Return” Despite Sustained Virologic 

Response? 

Meera Ramanathan 1 , David W. Backstedt 2 , Mark Pedersen 1 , 

Myunghan Choi 4 , Anil B. Seetharam 3 ; 1 Internal Medicine, Banner 

University Medical Center, Phoenix, AZ; 2 Gastroenterology, 

Banner University Medical Center, Phoenix, AZ; 3 Banner Transplant 

and Advanced Liver Disease Center, University of Arizona 

College of Medicine-Phoenix, Phoenix, AZ; 4 College of Nursing 

and Healthcare Innovation, Arizona State University, Phoenix, AZ 

Rationale: Direct acting antiviral (DAA) regimens for Hepatitis 

C (HCV) achieve high sustained virologic response (SVR12) 

rates. Abrogation of fibrosis is a well described salutary effect 

of HCV eradication in non-cirrhotic populations; however beneficial 

effects in those with cirrhosis require further analysis. 

Aims: Evaluate SVR12 rate in a cohort of cirrhotic patients and 

quantify change in laboratory parameters with DAA therapy. 

Secondary: assess factors that predict magnitude or direction 

of change in model for end stage liver disease (MELD) score 

in cirrhotics achieving SVR12. Methods: Prospective cohort 

study of consecutive mono-infected cirrhotic patients initiated 

on DAA regimen: 1) pegylated interferon alfa 2a (IFN) + 

Sofosbuvir (SOF) + Ribavirin (RBV); 2) SOF + RBV; 3) SOF 

+ Simeprevir (SMV); 4) Ledipasvir (LDV) + SOF. Baseline 

demographics tabulated: gender, ethnicity, genotype, DAA 

regimen, prior treatment, and prevalence of pre-existing: diabetes, 

hypertension, and hyperlipidemia. Laboratory parameters 

recorded at start, end, and assessment of SVR12 including: 

MELD components/score and platelet count. Changes in lab 

values compared between groups that did or did not achieve 

SVR12 using student’s T test. Logistic regression incorporated 

baseline demographics and calculated adjusted odds ratios 

for a decrease in MELD with treatment (p


1146 

Efficacy of Ledipasvir plus Sofosobuvir with or without 

ribavirin in hepatitis C genotype 1 patients who failed 

previous treatment with Simeprevir plus Sofosbuvir 

Gabriel R. Gonzales 3 , Stevan A. Gonzalez 1 , Hector E. Nazario 2 , 

Manjushree Gautam 1 , Maisha Barnes 2 , Parvez S. Mantry 2 , Jeffrey 

S. Weinstein 2 , Milka Ndungu 2 , Olga Teachenor 2 , Apurva A. 

Modi 1 ; 1 Liver Consultants of Texas, Fort Worth, TX; 2 Liver Institute 

at Methodist-Dallas, Dallas, TX; 3 Texas College of Osteopathic 

Medicine, University of North Texas HSC, Fort Worth, TX 

Background: Combination therapy of Simeprevir (SIM), NS3/4 

protease inhibitor, with Sofosbuvir (SOF), NS5B polymerase 

inhibitor is an FDA approved treatment option for chronic hepatitis 

C genotype 1 patients with an overall SVR12 rate of 

85-95%. Single tablet fixed dose combination of Ledipasvir 

(LDV), NS5A inhibitor, with SOF is also FDA approved for 

treatment of hepatitis C genotype 1 with SVR 12 rates of ≥ 

95%. However, there is no data on the efficacy of re-treatment 

with LDV+SOF in patients who failed initial treatment 

with SIM+SOF. Aim: To evaluate the efficacy of re-treatment 

with LDV+SOF with or without ribavirin (RBV) in genotype 1 

patients who have previously failed treatment with SIM+SOF. 

Methods: Data from a combined treatment cohort of two large 

hepatology referral centers, which included patients who were 

previously treated with SIM+SOF with or without RBV for 12 

weeks but failed to achieve SVR12 & then undergone re-treatment 

with LDV+SOF with or without RBV was analyzed (n=28). 

LDV+SOF with or without ribavirin was administered for 12-24 

weeks based on the discretion of the treating hepatologist. 

Results: Of the 28 patients in this cohort, 21 (75%) were male, 

74% were Caucasians & 24 (86%) were genotype 1A. 22 

(79%) had cirrhosis of which 54% had decompensated Child 

class B/C cirrhosis. Three patients had recurrent hepatitis C 

post-liver transplant. 21/28 (75%) achieved undetectable viral 

load after 4 weeks of treatment. Of the 3 patients who have 

currently completed treatment, all achieved undetectable viral 

load at the end of treatment (EOT). Treatment was well tolerated 

overall with over 38% reporting no adverse events. In 

those reporting adverse events, the most common were fatigue, 

headache, insomnia, nausea, & diarrhea. No patients with 

decompensated child class B/C cirrhosis had further decompensation 

requiring hospitalization or death during treatment. 

SVR12 results to follow. Conclusions: Single tablet fixed dose 

combination of Ledipasvir plus Sofosbuvir with or without ribavirin 

is well tolerated in patients who previously failed treatment 

with Simeprevir plus Sofosbuvir including those with decompensated 

cirrhosis & recurrent hepatitis C post-liver transplant. 

Overall, 78% have achieved negative viral load after 4 weeks 

of treatment & all (n=3) who have reached EOT have undetectable 

viral load. 

The following authors have nothing to disclose: Gabriel R. Gonzales, Maisha 

Barnes, Milka Ndungu, Olga Teachenor 

1147 

Safety And Efficacy Of Two IFN- Free Daas Regimens 

In Genotype 4 Chronic HCV Patients: First Real Clinical 

Practice Data In Gulf From Qatar HCV Registry 

Moutaz F. Derbala 1 , Aliaa Amer 2 , Saad R. Alkaabi 1 , Yasser M. 

Kamel 1 , Khaleel H. Sultan 1 , Elham Elsayad 3 , Omer E. Hajelssedig 1 , 

Nazeeh Z. Dweik 1 , Mohammed T. Butt 1 , Mohammed E. Elbadri 1 ; 

1 GASTROENTEROLOGY & HEPATOLOGY, Hamad Hospitaland 

Weill Cornell Medical College in Qatar,Consultant Gastroenterology 

& Hepatology HMC, Doha, Qatar; 2 Laboratory Medicine and 

Histopathology, Hematology Section, Doha, Qatar., Hamad Medical 

Corporation, Doha, Qatar; 3 Clinical Pharmacology, Hamad 

Medical Corporation, Doha, Qatar 

Background and Aims: IFN-free regimens of direct acting antivirals 

(DAAs): sofosbuvir (SOF), simeprevir (SMV), and daclatasvir 

(DCV) with or without ribavirin (RBV) have been used in 

Qatar. In real world; little data are currently available on use 

of DAAs in genotype 4 HCV patients including patients with 

advanced liver diseases and comorbidities in gulf. Our aim is 

to assess efficacy and safety of 12 weeks regimen of SOF/SMV 

or SOF/DCV versus 24 weeks regimen of SOF/RBV in genotype 

4 HCV patients. Methods: 85 chronic HCV genotype 4 

patients received either 12 weeks of SOF/SMV or SOF/DCV, 

or 24 weeks of SOF plus ribavirin (RBV) therapy. 30(35.3%) 

and 10(11.8%) patients were treated with SOF/SMV and 

SOF/DCV for 12 weeks respectively, while 45(52.9%) patients 

were treated with SOF/RBV for 24 weeks. Demographics, history 

of liver disease, laboratory tests, adverse events and virological 

data were collected at baseline, throughout treatment 

and post-treatment follow-up. Results: 34.3% of patients had 

cirrhosis and 58.8% failed on previous PR treatment. 12.9% 

of patients underwent liver transplantation. (Table) To date 

84/85 patients continued treatment on both regimen, while 

1 patient on SOF/DCV discontinued voluntarily. The SVR4 is 

96%, with 1 virological breakthrough due to noncompliance 

and 2 relapses in experienced patients were reported in the 

12 weeks regimen group of SOF/SMV or DCV. No serious 

adverse events were reported in any treatment groups, while 

26% of the SOF/RBV group reported grade 3 or 4 anemia 

which are managed by RBV dose reduction ; with 2 patients 

received erythropoietine and 1 patient received blood transfusion. 

Conclusion: In Genotype 4 chronic HCV patients, both 

12 weeks regimen of SOF in combination with SMV or DCV 

and 24 weeks regimen of SOF/RBV achieved high SVR in real 

world setting. However, the shorter duration of 12 weeks of 

SOF/SMV or DCV and the favorable tolerability and safety 

profile make this regimen more recommended in Genotype 4. 

Disclosures: 

Stevan A. Gonzalez - Consulting: AbbVie; Speaking and Teaching: Gilead, 

Salix, AbbVie 

Hector E. Nazario - Speaking and Teaching: Merck, Salix 

Manjushree Gautam - Speaking and Teaching: Gilead 





Salix 

Jeffrey S. Weinstein - Speaking and Teaching: Merck 

Apurva A. Modi - Speaking and Teaching: Salix, Merck, Gilead, Abbvie, Janssen 

Disclosures: 

The following authors have nothing to disclose: Moutaz F. Derbala, Aliaa Amer, 

Saad R. Alkaabi, Yasser M. Kamel, Khaleel H. Sultan, Elham Elsayad, Omer 

E. Hajelssedig, Nazeeh Z. Dweik, Mohammed T. Butt, Mohammed E. Elbadri


1148 

Preliminary Experience Of Direct Acting Antiviral Therapy 

In Hepatitis C Infected Kidney Transplant Recipients, 

Who Received Grafts From Hepatitis C Positive Or Negative 

Donors 

Kalyan R. Bhamidimarri, David Roth, Giselle Guerra, Cynthia Levy, 

Paul Martin; University of Miami-Miller School of Medicine, Miami, 

FL 

Background: Highly effective direct acting antiviral therapy 

(DAA) is now feasible post kidney transplantation (KT) and has 

also facilitated a new strategy of abbreviating time in waitlisted 

patients who could accept Hepatitis C (HCV) positive 

grafts. However there are no reports of treatment experience 

in this group to date. Aim: We describe our preliminary experience 

of HCV treatment with DAA’s in post KT patients, a 

majority of whom received a graft from HCV+ donor. Methods: 

Fourteen patients with chronic HCV, genotype 1, 71% males, 

57% African Americans, mean age of 54 years underwent 

deceased donor kidney transplantation (71% HCV+ graft). 

Baseline features include 86% with genotype 1a, 50% with 

fibrosis level greater than F2, mean HCV viral load of 3.4 

Million IU, mean hemoglobin of 12gm/ dL and mean GFR of 

60mL/ minute. Open label treatment with Sofosbuvir / Ledipasvir 

was initiated in 13 patients and one patient received 

Sofosbuvir plus Simeprevir. Only 65% of the patients had ribavirin 

approved in their regimen and the dosage was titrated 

according to their tolerance. Treatment was initiated within 

a mean period of 5months since the KT (range 1-10months) 

and immunosuppression was closely monitored. Results: None 

of the patients developed fibrosing cholestatic hepatitis post 

KT, despite receiving a graft from HCV+ donor. All patients 

are at various stages of completion of their respective treatment 

duration. Five patients reached end of treatment (EOT), 2 

patients reached post-treatment week 4 and 1patient reached 

post treatment week 12. According to per protocol analysis, 

the EOT, SVR4 and SVR12 rates are 100%. Anemia was frequent 

in the patients taking ribavirin which warranted frequent 

dose titrations. Conclusions: Our preliminary experience shows 

an evolving successful strategy of transplanting kidneys from 

HCV+ donor in HCV infected recipients and the feasibility of 

HCV treatment with newer DAA’s. As per protocol analysis, 

all the patients had excellent virologic response and full data 

awaited by AASLD Liver Meeting. 

HCV Treatment in KT recipients 

Disclosures: 

Kalyan R. Bhamidimarri - Advisory Committees or Review Panels: Gilead, Abb- 

Vie, Janssen; Grant/Research Support: Bristol Squibb Myers, Biotest, Synageva, 

Salix, Vital Therapies, Ocera Inc 

David Roth - Advisory Committees or Review Panels: Merck, Sharp and Dome; 

Consulting: Merck, Sharp and Dome 



The following authors have nothing to disclose: Giselle Guerra, Cynthia Levy 

1149 

IFNL4 ss469415590 Polymorphism Contributes to Treatment 

Decisions in Patients with Chronic Hepatitis C virus 

Genotype 1b but not 2a Infection 

Ruihong Wu 1 , Xiumei Chi 1 , Haibo Sun 1 , Juan Lv 1 , Xiaomei 

Wang 1 , Xiuzhu Gao 1 , Ge Yu 1 , Fei Kong 1 , Peng Zhang 2 , Mo-li 

Wang 3 , Dongsheng Wang 4 , Tao Jiang 4 , Hongqin Xu 1 , Rui Hua 1 , 

Gerald Y. Minuk 5 , Jing Jiang 6 , Bing Sun 7 , Jin Zhong 7 , Yu Pan 1 , 

Junqi Niu 1 ; 1 Department of Hepatology, First hospital of JiLin University, 

ChangChun, China; 2 Department of Infectious Diseases, 

First hospital of Jilin University, Changchun, China; 3 Department of 

Infectious Diseases, Fourth Hospital of Jilin University, Changchun, 

China; 4 Hospital of HepatologyBiliary of Jilin Province, Changchun, 

China; 5 Section of Hepatology, University of Manitoba, Winnipeg, 

MB, Canada; 6 Department of Clinical Epidemiology, First 

Hospital of Jilin University, Changchun, China; 7 Institute Pasteur 

of Shanghai, Shanghai Institutes for Biological Sciences, Chinese 

Academy of Sciences, Shanghai, China 

Background & Aim: Recently, the dinucleotide variant 

ss469415590(TT/ΔG) in the novel gene IFNL4 was identified 

as a stronger predictor of hepatitis c virus (HCV) clearance in 

individuals of African ancestry compared with rs12979860. 

The aim of this study was to determine whether this variant 

contributes to treatment decisions in a Chinese population, 

predominantly infected with HCV genotype (GT) 1b and 2a. 

Methods: Four hundred and forty seven chronic hepatitis c 

(CHC) patients (including 328 interferon alpha-2b and ribavirin 

treated), 129 individuals who had undergone spontaneous 

HCV clearance (SHC) and 169 healthy controls were 

retrospectively investigated. ss469415590 genotyping was 

performed using a Mass spectra method (SEQUENOM). 

Results: A higher proportion of SHC individuals carried TT/TT 

genotype compared to CHC patients (95.3% vs. 88.8%, P = 

0.027). ss469415590 variant was independently associated 

with sustained virological response (SVR, OR = 3.247, 95% 

CI = 1.038-10.159, P = 0.043) and on-treatment virological 

responses including rapid virological response, complete early 

virological response, early virological response and end of 

treatment virological response with a minimal OR of 3.73. 

Patients with high viral load (≥4×10 5 IU/mL) and the ΔG allele 

had a higher risk of failing to achieve SVR compared to TT/ 

TT genotype (92.9% vs. 64%, P = 0.034). In GT-2a patients, 

no significant association of ss154949590 variant with virological 

response was identified. In addition, we found that 

ss154949590 was in complete linkage disequilibrium with 

rs12979860. Conclusions: IFNL4 ss154949590 TT/TT genotype 

favours spontaneous clearance of HCV, and treatment 

induced clearance in genotype 1b but not 2a infected patients. 

IFNL4 ss469415590 (or rs12979860) genotyping should 

be considered for patients with genotype 1b and high viral 

load when making a choice between standard dual therapy 

and IFN-free direct-acting antiviral regimen. If with the ΔG for 

ss469415590 (or T for rs12979860), IFN-free DAA regimens 

may be better options. 

Disclosures: 

The following authors have nothing to disclose: Ruihong Wu, Xiumei Chi, Haibo 

Sun, Juan Lv, Xiaomei Wang, Xiuzhu Gao, Ge Yu, Fei Kong, Peng Zhang, Mo-li 

Wang, Dongsheng Wang, Tao Jiang, Hongqin Xu, Rui Hua, Gerald Y. Minuk, 

Jing Jiang, Bing Sun, Jin Zhong, Yu Pan, Junqi Niu


1150 

Combination therapies with daclatasvir and asunaprevir 

on NS3-D168 mutated hepatitis C virus in human 

hepatocyte chimeric mice 

Hiromi Kan 1 , Michio Imamura 1 , Nobuhiko Hiraga 1 , Eisuke 

Miyaki 1 , Takuro Uchida 1 , Masataka Tsuge 1 , Hiromi Abe 1 , Nelson 

Hayes 1 , Hiroshi Aikata 1 , Chise Tateno 2 , Kazuaki Chayama 1 ; 

1 Department of Gastroenterology and Metabolism, Applied Life 

Sciences, Institute of Biomedical & Health Sciences, Hiroshima 

University, Hiroshima, Japan; 2 PhoenixBio Co., Ltd., Higashihiroshima, 

Japan 

Background and Aim: The frequency of emergent drug-resistant 

strains of hepatitis C virus (HCV) in patients who failed to 

respond to simeprevir plus peginterferon (PEG-IFN) and ribavirin 

(RBV) combination therapy decreased after cessation of 

the treatment. However, it is not clear whether or not the HCV 

NS3-D168 mutation affects the outcome of an IFN-free combination 

therapy including NS5A inhibitor, daclatasvir and NS3 

protease inhibitor, asunaprevir. In this study, we investigated 

the relationship between the effect of daclatasvir plus asunaprevir 

treatment and the frequencies of drug-resistant NS3-D168 

variants using HCV-infected mice. Methods: Injection with wildtype 

and NS3 D168 mutated genotype 1b HCV mixed serum 

to human hepatocyte chimeric uPA-SCID mice enabled us to 

develop mice with various frequencies of NS3-D168 mutation. 

These mice were treated with 40 mg/kg of asunaprevir alone or 

in combination with 10 mg/kg daclatasvir for four weeks (provided 

by Bristol-Meyers Squibb Research and Development). 

Frequencies of NS3-D168 mutation at baseline were analyzed 

by ultra-deep sequencing. Some mice infected with NS3-D168 

substitutions were administered with either intramuscular injection 

of 30 μg/kg of PEG-IFN-alfa twice a week or 200 mg/kg 

of telaprevir (provided from Mitsubishi Tanabe Pharma) orally 

twice per day for four weeks. Results: Mice with high NS3- 

D168 variant frequencies showed a low susceptibility to asunaprevir 

mono-therapy and failed to respond to daclatasvir plus 

asunaprevir therapy. In contrast, mice with a low frequency 

(less than approximately 14%) of NS3-D168 variants showed 

a similar susceptibility to asunaprevir mono-therapy with wildtype 

HCV-infected mice and achieved viral eradication with 

four weeks of daclatasvir plus asunaprevir therapy. Although 

telaprevir or PEG-IFN treatment resulted in a reduction of serum 

HCV RNA levels, no significant decrease in the frequency of 

NS3-D168 substitutions was achieved. Conclusion: Daclatasvir 

and asunaprevir treatment could eliminate NS3-D168-mutated 

HCV if the frequency at baseline was low. Using either telaprevir 

or IFN might cause no significant reductions in NS3- 

D168 mutation. It is necessary to confirm that the frequency of 

NS3-D168 variants has decreased sufficiently before adopting 

daclatasvir plus asunaprevir therapy in patients with simeprevir 

plus PEG-IFN/RBV treatment failure. 

Disclosures: 








The following authors have nothing to disclose: Hiromi Kan, Michio Imamura, 

Nobuhiko Hiraga, Eisuke Miyaki, Takuro Uchida, Masataka Tsuge, Hiromi Abe, 

Nelson Hayes, Hiroshi Aikata, Chise Tateno 

1151 

HepCure: An Innovative web-based toolkit to train a 

new cohort of hepatitis C providers and increase patient 

engagement 

Ponni V. Perumalswami 1 , Korin Parrella 2 , Jason Rogers 3 , Rahul 

Patel 4 , Brooke Wyatt 1 , Kian Bichoupan 1 , Andrea D. Branch 1 , Brian 

Kim 1 , Anna Patel 1 , Joseph Lawler 1 , Alicia Stivala 2 , Alyson Harty 1 , 

Donald Gardenier 2 , Michel Ng 1 , Catherine Amory 2 , Aaron M. 

Vanderhoff 1 , Douglas Dieterich 1 , Ashish Atreja 3 , Jeffrey J. Weiss 2 ; 

1 Liver Diseases, Icahn School of Medicine at Mount Sinai, New 

York, NY; 2 Medicine, Icahn School of Medicine at Mount Sinai, 

New York, NY; 3 Medicine- Division of Gastroenterology, Icahn 

School of Medicine at Mount Sinai, New York, NY; 4 Parsus Solutions, 

LLC, Scottsdale, AZ 

Background and Aims: There is a great need to expand the 

number of health care professionals treating hepatitis C and 

to optimize patient engagement. The aim of this study is to 

develop and operationalize HepCure (http://hepcure.org), 

a web-based application (app) with three components: An 

open access toolkit (a dashboard) that enhances providers’ 

ability to deliver guideline-based HCV care; a patient app that 

provides education, medication reminders, and a platform 

for tracking adherence and symptoms; and a tele-education 

platform for medical providers. Methods: The HepCure app 

is a collaboration among an academic, tertiary care medical 

center, the New York State (NYS) Department of Health, 

and NYS community health centers. A working group of multidisciplinary 

HCV experts (Hepatology, Internal Medicine, 

Psychology, Social Work, and Public Health) was created to 

develop content for the HepCure dashboard. Input from this 

group was used to develop wireframes. Focus groups were 

held with providers and patients to refine wireframes. National 

guidelines (AASLD/IDSA) were used to identify key patient 

characteristics to determine treatment recommendations to 

build decision support algorithms. The app has the capability 

to de-identify patient data to directly discuss cases selected 

by providers at weekly tele-education sessions with experts. 

Results: The HepCure platform captures United States Centers 

for Medicaid and Medicare and NYS HCV quality indicators 

for population health management, provides decision support 

algorithms for providers that can be updated in real time with 

newly approved HCV treatment regimens and incorporates 

a tele-education platform with access to experts. The Hep- 

Cure Provider Dashboard (http://providers.hepcure.org) was 

launched in November 2014. Weekly tele-education sessions 

have been conducted since February 2015 (n=17) and each 

session has been archived with open access. These sessions 

have been attended by an average of 13 ± 7 attendees a 

week and archived sessions have been viewed an average of 

8 ± 3 times. A patient mobile app which can be linked to the 

provider dashboard was launched in June 2015 to increase 

patient engagement. Conclusions: Providers are interested in 

web-based tools to help them master HCV treatment and stay 

current with ongoing changes in treatment paradigms. Patients 

with HCV desire web-based tools to help them learn more 

about and navigate HCV treatment. The HepCure platform is 

an innovative model that can be adapted globally in line with 

local HCV treatment guidelines to train a new generation of 

HCV providers, collect real world data, and foster comparative 

effectiveness research. 

Disclosures: 

Kian Bichoupan - Consulting: Janssen Pharmaceuticals, Gilead Sciences 



Jannsen, Acaria Pharmacy, Abbvie


Michel Ng - Advisory Committees or Review Panels: abbvie; Speaking and 

Teaching: abbvie 



Jeffrey J. Weiss - Consulting: AbbVie Inc. 

The following authors have nothing to disclose: Ponni V. Perumalswami, Korin 

Parrella, Jason Rogers, Rahul Patel, Brooke Wyatt, Brian Kim, Anna Patel, Joseph 

Lawler, Alicia Stivala, Donald Gardenier, Catherine Amory, Aaron M. Vanderhoff, 

Ashish Atreja 

1152 

Patient Expectations of the Impact on Quality of Life of 

Hepatitis C Treatment 

Shelagh M. Szabo 2 , David R. Walker 1 , Timothy R. Juday 1 , 

Suzanne Lane 2 , Adrian Levy 2 , Sammy Saab 3 ; 1 HEOR, Abbvie, 

North Chicago, IL; 2 Icon Health Economics, Vancouver, BC, Canada; 

3 David Geffen School of Medicine, UCLA, Los Angeles, CA 

BACKGROUND The Investigative Multicenter Prospective Observational 

Study (IMPROOV) is a study of humanistic and economic 

outcomes among patients initiating direct acting antiviral 

treatments for hepatitis C virus (HCV). The objective of this analysis 

is to describe patient-reported expectations of the impact 

on quality of life of newly-initiated HCV treatment. METHODS 

All consecutive HCV patients attending clinics at ten sites in 

the United States were screened to identify the cohort initiating 

treatment for HCV. Data were collected using patient-completed 

surveys at up to five time points over the treatment 

course, supplemented by a medical chart review at treatment 

initiation and completion. Data collected included demographic, 

clinical, economic, and humanistic. Data on patients’ 

expectations of the impact on quality of life of newly-initiated 

treatment were collected; multiple responses were allowed per 

patient. The specific question asked at baseline was: “Do you 

expect the following to occur, when you start HCV treatment”; 

and a series of potential patient experiences were listed to 

which patients responded with the expected impact. Response 

options included: none, a little, moderate or severe. The sum 

of patients expecting moderate/severe outcomes was calculated. 

First patient enrolled 1/2013 and last patient enrolled 

9/2014. RESULTS 143 patients, from 8 clinics, enrolled out 

of 787 that were screened across 10 clinics of which 137 

were from community clinics and 6 from academic/hospitals. 

Mean age was 56 years, 52% male, 60% white, 20% hispanic 

and 15% black. 75% graduated from high school and 

33% were employed. 100 patients were on a sofosbuvir and/ 

or simeprevir-based regimen, and 43 patients were on other 

treatments. 74 were on an interferon-free (IFF) regimen. The 

proportion of patients expecting moderate/severe impacts of 

newly initiated treatment for those on IFF and interferon-containing 

treatment, respectively, were: treatment-related side-effects 

(31.5%, 50.7%;p=0.02), missed work (7.2%,17.9%;p=0.06 

), decreased productivity at work (9.9%, 25.0%;p=0.03), limited 

social activities (25.0%,40.6%;p=0.05), increased reliance 

on family/friends (18.8%,36.2%;p=0.02), impact on 

your quality of life (21.4%,49.3%;p


Concluding, the combined use of IFNL4 polymorphisms and 

ALT reduction at 4 week of treatment is able to optimize candidates’ 

selection for antiviral therapy with PEG-IFN and RBV in 

patients with early liver fibrosis, discriminating those that could 

still benefit from dual therapy from the ones that will need the 

new regimens. 

Disclosures: 


Jannsen 

The following authors have nothing to disclose: Francesco Figorilli, Simona 

Onali, Stefania Catone, Claudio Argentini, Stefania Casu, Cinzia Balestrieri, 

Maria Conti, Giancarlo Serra, Michele Casale, Maria Cristina Pasetto, Laura 

Matta, Lucia Barca, Rosetta Scioscia, Irene Canini, Maria Giovanna Quaranta, 

Domenico Genovese, Stefano Vella 

1154 

Real Life Treatment Outcomes with 8 week Course 

Treatment for Hepatitis C without Cirrhosis Confirmed 

by Transient Elastography 

Vanessa Marshall 2,3 , Kelsey Rife 1 , Amy Hirsch 1,3 , Marina G. Silveira 

2,3 , Anita Compan 2 , Anita L. Moreland 2 , Yngve Falck-Ytter 2,3 ; 

1 Pharmacy, Louis Stokes Cleveland VA Medical Center, Cleveland, 

OH; 2 Gastroenterology Section, Louis Stokes Cleveland VA Medical 

Center, Cleveland, OH; 3 School of Medicine, Case Western 

Reserve University, Cleveland, OH 

Background: Short course treatments for Hepatitis C (HCV) 

are less expensive for payers and easier for patients. Data is 

needed to confirm that real world sustained virologic response 

(SVR) rates for short courses of treatment in patients deemed 

non-cirrhotic based on transient elastography (TE, FibroScan) 

results are comparable to SVR rates seen in registrational trials. 

It is also not known if cutoffs from non-invasive markers of 

liver fibrosis assessment (Fibrosis-4 score (FIB-4) or aspartate 

transaminase to platelet ratio index (APRI) score) are sufficient 

to select for short course treatment. Objective: This study was 

designed to examine (1) if a TE score < 12.5 kPa is sufficient 

criteria to rule out cirrhosis in candidates eligible for 8 weeks 

of ledipasvir/sofosbuvir; and (2) evaluate if the clinical cutoffs 

for FIB-4 (< 3.25) and APRI score (< 1) are sufficient criteria 

to shorten the course of treatment. Methods: Veteran genotype 

1 (GT1), treatment naïve and previous interferon/ribavirin 

relapsers with pre-treatment TE scores of < 12.5 kPa and HCV 

viral loads < 6 million IU/mL were treated with 8 weeks of 

ledipasvir/sofosbuvir. Pre-treatment FIB-4 and APRI scores were 

calculated. Outcomes were assessed at 4 (SVR4), 12 (SVR12), 

and 24 (SVR24) weeks post-treatment. Results: To date, 52 Veterans 

were initiated on an 8 week course of ledipasvir/sofosbuvir. 

94% were male with a mean age of 61 (range 32-75). 6 

patients were previous treatment relapsers with interferon/ribavirin. 

31 patients (60%) had GT1a, 18 (35%) had GT1b and 

3 (5%) had GT1 (no subtype) or GT 1a/1b. 41 patients (78%) 

had baseline TE scores < 9.5 kPa. To date, 17/18 (94%) 

patients achieved SVR4 and 1 patient relapsed. 5/17 (29%) 

patients who achieved SVR4 had an APRI ≥ 1 and/or FIB-4 ≥ 

3.25. The patient who relapsed after 8 weeks of ledipasvir/ 

sofosbuvir was treatment naïve, GT1a, had a TE score of 11.2 

kPa, and APRI > 1 but FIB-4 < 3.25. Viral loads were undetectable 

at week 4 and end of treatment and the patient reported 

no missed doses of ledipasvir/sofosbuvir. Final SVR12 results 

will be presented for all 52 patients. Conclusions: A treatment 

strategy using TE to qualify patients for an 8 week course of 

ledipasvir/sofosbuvir resulted in similar SVR rates to those seen 

in registrational trials. These results suggest a TE score < 12.5 

kPa is sufficient criteria to shorten the course of HCV treatment 

to 8 weeks with > 90% SVR rate. Due to lower diagnostic accuracy 

of APRI and FIB-4 scores, their application at suggested 

cutoffs may result in an additional 4 weeks of ledipasvir/sofosbuvir, 

resulting in unnecessary resource expenditure. 

Disclosures: 

The following authors have nothing to disclose: Vanessa Marshall, Kelsey Rife, 

Amy Hirsch, Marina G. Silveira, Anita Compan, Anita L. Moreland, Yngve 

Falck-Ytter 

1155 

Effectiveness of Treatment of Chronic Hepatitis C in 

Patients Being Treated for Hepatocellular Cancer: A Single 

Center Experience 

Shruti Mony 2 , Shirin Nafisi 1 , Justin A. Reynolds 1 , Ann Moore 1 , 

Ashley Hepner 1 , Karen Arendt 1 , Yvette Cummings 1 , Robert Gish 1 , 

Richard A. Manch 1 , Anita Kohli 1 ; 1 Hepatology, St. Joseph’s Hospital 

and Medical Center, Phoenix, AZ; 2 Internal Medicine, St. 

Joseph’s Hospital and Medical Center, Phoenix, AZ 

Treatment of chronic hepatitis C (CHC) with directly acting 

antivirals has improved SVR rates for patients with advanced 

liver disease. Patients with hepatocellular cancer (HCC) remain 

a group with a poor prognosis. Effectiveness of CHC treatment 

in this group, often receiving other anti-cancer agents or therapies, 

is unknown. METHODS: Patients treated for CHC during 

or after HCC treatment at a community based academic medical 

center, between 1/12- 5/15 were retrospectively identified 

and included. Demographics, CHC and HCC treatment, 

virologic data, treatment outcomes, HCC stage, and survival 

were collected. RESULTS: 35 patients with HCC treated for 

CHC concomitantly or after HCC therapy were identified. 62% 

were men, 88% Caucasian, 17% Hispanic and 54% treatment 

experienced. 22 patients were treated for HCC and CHC 

concomitantly. In patients with GT1, 12 received ledipasvir 

(LDV)/sofosbuvir (SOF) for 24 weeks: 8 (single HCC) received 

Y90/microwave ablation/sorafenib, Y90, TACE, RFA/TACE/ 

Y90, microwave ablation, TACE/quadsphere or await liver 

transplant; 3 (multifocal HCC) received TACE/sorafenib, 

TACE or TACE/RFA/radiation for prostate adenocarcinoma; 

1 (metastatic HCC)received Y90. 1 patient (metastatic HCC) 

received LDV/SOF for 12 weeks with embolization/ sorafenib. 

4 patients received SOF/simeprevir (SMV) for 12 weeks with 

TACE, TACE/ Y90/transplant (single HCC), cyberknife/ 

sorafenib or TACE/sorafenib (metastatic HCC). 1 patient (GT2 

& metastatic HCC) received SOF/ribavirin (RBV) for 20 weeks 

& Y90/TACE/sorafenib. 4 patients (GT3) received SOF/RBV 

for 24 weeks with Y90/TACE/transplant or microablation 

(single HCC) or TACE/sorafenib or TACE(multifocal HCC). In 

patients who reached SVR 12, 50% (n=3) achieved SVR 12, 

33% (n=2) relapsed & 17% (n=1) died. SVR12 is pending 

for 16 patients. 13 patients were treated for CHC after HCC 

therapy. 3 (GT1,single HCC) received Y90/microablation/ 

RFA or Y90 followed by LDV/SOF for 12 weeks. 6 (GT1, 

single HCC) received TACE, RFA, microablation or TACE/Y90 

followed by SOF/SIM for 12 weeks. 2 (GT2) received TACE 

for single & multiple HCC followed by SOF/RBV for 12 weeks. 

1 patient (GT3) received Y90 for a single HCC followed by 

SOF/RBV for 24 weeks. In patients who reached SVR 12, 

70% (n=7) achieved SVR 12 & 30% (n=3) relapsed. SVR12 is 

pending for 3 patients. Full results of SVR12 will be presented. 

CONCLUSION: Early results suggest that the treatment of CHC 

in patients with HCC, particularly those undergoing concomitant 

HCC therapy may be lower than other advanced liver 

disease patients. Immune modulating agents and liver targeted 

therapies may reduce the effectiveness of HCV interferon free 

therapies.


Disclosures: 





The following authors have nothing to disclose: Shruti Mony, Shirin Nafisi, Justin 

A. Reynolds, Ann Moore, Ashley Hepner, Karen Arendt, Yvette Cummings, Anita 

Kohli 

1156 

Sofosbuvir-based treatments for patients with hepatitis 

C virus (HCV) mono-infection and human immunodeficiency 

virus (HIV)-HCV co-infection with genotype 1 and 

4 in clinical practice – Results from the GErman hepatitis 

C COhort (GECCO) 

Stefan Mauss 1 , Knud Schewe 2 , Jürgen K. Rockstroh 3 , Dietrich 

Hueppe 4 , Axel Baumgarten 5 , Guenther Schmutz 1 , Karl-Georg 

Simon 6 , Thomas Lutz 8 , Heiner W. Busch 7 , Patrick Ingiliz 5 , Stefan 

Christensen 7 ; 1 Center for HIV and Hepatogastroenterology, 

Duesseldorf, Germany; 2 ICH, Hamburg, Germany; 3 Department I 

University Hospital Bonn, Bonn, Germany; 4 Center for Gastroenerology, 

Herne, Germany; 5 MIB, Berlin, Germany; 6 Practice for 

Gastroenterology, Leverkusen, Germany; 7 CIM, Muenster, Germany; 

8 Infektiologikum, Frankfurt, Germany 

Introduction: The first direct acting antivirals (DAA) were 

approved in Europe in 2014 based on limited study data. 

In particular, HIV-HCV co-infected patients and pre-treated 

patients had not been systematically studied. Here, we present 

real-life data on efficacy and safety of sofosbuvir (SOF)-based 

therapies from Germany. Methods: In this multicenter cohort, 

patients who were started on sofosbuvir-based treatments were 

documented consecutively. For the current analysis, patients 

included had HCV genotype 1 and 4 treated with: SOF/ 

PegIFN/RBV (ribavirin) 12 weeks, SOF/DCV (daclatasvir) 

12/24 weeks, SOF/SMV (simeprevir) 12 weeks and SOF/LDV 

(ledipasvir) 8/12 weeks. Patients within the GECCO cohort 

are part of the prospective German hepatitis C registry. Results: 

Overall, 836 patients on SOF-containing regimens have been 

enrolled so far. Of those, 629 (75%) are HCV-monoinfected 

and 207 (25%) HIV-HCV co-infected. 536 patients (60%) were 

male, the median age was 52 years. 421 patients (52%) had 

been pretreated, mainly with PegIFN and RBV. Liver cirrhosis 

was reported in 217 patients (27%), in the majority of cases 

(71%) defined by a transient elastography value (Fibroscan®) 

>12.5kPa. 17/217 (8%) cirrhotic patients had Child-Pugh 

stage B or C. HCV genotype 1 was present in 584 patients 

(69%), and genotype 4 in 48 patients (6%). Most HIV-HCV 

co-infected patients (205/207) were on antiretroviral therapy 

and had HIV-RNA 2) were less likely to achieve 

SVR12 compared to non-cirrhotic patients (APRI


sion] The combination therapy of DCV and ASV for patients 

with HCV genotype 1 had highly effectiveness. The efficacy 

and the safety of this therapy for old patients were equal to the 

therapy for young patients. NS5B polymerase inhibitor sofosbuvir 

is not available for patients with severe renal dysfunction. 

We may positively select the combination therapy DCV and 

ASV for elderly patients, because elderly patients tended to 

have decreased renal function. 

Disclosures: 

The following authors have nothing to disclose: Kenta Motomura, Masayoshi 

Yada, Taiji Mutsuki, Masayuki Miyazaki, Takeshi Senju, Motoyuki Kohjima, 

Makoto Nakamuta, Akihide Masumoto 

1158 

Sofosbuvir-based antiviral therapy in HCV patients with 

severe renal failure 

Jérôme Dumortier 1 , François Bailly 2 , Georges-Philippe Pageaux 3 , 

Anaïs Vallet-Pichard 4 , Sylvie Radenne 2 , François Habersetzer 5 , 

Marie-Claude Gagnieu 1 , Jean Didier Grange 6 , Anne Minello 7 , 

Nassim Kamar 8 , Laurent Alric 9 , Vincent Leroy 10 ; 1 Hôpital Edouard 

Herriot, Lyon, France; 2 Croix-Rousse Hospital, Lyon, France; 3 Saint 

Eloi University Hospital, Montpellier, France; 4 Cochin Hospital, 

Paris, France; 5 University Hospital of Strasbourg, Strasbourg, 

France; 6 Tenon Hospital, Paris, France; 7 Dijon University Hospital, 

Dijon, France; 8 Rangueil University Hospital, Toulouse, France; 

9 Purpan University Hospital, Toulouse, France; 10 CHU A Michallon, 

Grenoble, France 

Background: Chronic hepatitis C virus (HCV) infection is the 

most common chronic liver disease in patients with end stage 

renal disease (ESRD). During the last year, second generation 

direct acting antivirals have been a revolution for the treatment 

of hepatitis C, and sofosbuvir (SOF) is the cornerstone 

of modern treatment. Since SOF is eliminated through kidney, 

the aim of this multicentre retrospective study was to assess its 

antiviral efficacy and tolerability in HCV infected patients with 

severe renal failure (including haemodialysed). Patients: Fifty 

patients (36 male, mean age 60.5±7.5 years) with chronic 

HCV-infection (7 GT1a, 21 GT1b; 6 GT2; 5 GT3; 9 GT4; 2 

GT5; cirrhosis: 27/54%) with severe renal failure, i.e. GFR 

< 35 mL/min, including 35 haemodialysed, were included 

in this study. There were 17 patients with history of kidney 

transplantation, 11 patients with history of liver transplantation, 

and 27 patients were on waiting list for kidney transplantation. 

Fourteen patients were naïve of antiviral treatment. Antiviral 

treatment consisted in SOF/ribavirin (RBV) (n=7), SOF/RBV/ 

PEG-IFN (n=2), SOF/daclatasvir (DCV) ±RBV (n= 30), or SOF/ 

simeprevir (SMV) ±RBV (n= 11). Treatment duration was 12 or 

24 weeks according to regimen. Reduced dose of SOF (400 

mg 3 times a week or 400 mg every other day) was given 

in all haemodialysed patients. Results. On-treatment response 

(rate of undetectable RNA) was as follows: week 4 : 36/44 

(82%), week 8 : 38/40 (95%), week 12: 50/50 (100%), 

week 24 : 10/10 (100%). Sustained virological response rate 

was 27/29 (93%) at 4 weeks (SVR4) and 24/26 (92%) at 12 

weeks (SVR12). The 2 patients with virological relapse were 

both G3, with severe fibrosis, who were treated for 12 weeks 

without RBV. Dose of RBV (n=13) ranged from 200 mg 3 times 

a week to 600 mg/day. The mean baseline hemoglobin level 

was 11.6±1.4 g/dL and 19 patients were on Epoetin before 

starting antiviral therapy. At the end of treatment, the mean 

hemoglobin level was 11.7±1.8 g/dL and 19 patients were on 

Epoetin. Hemoglobin level was not different in patients receiving 

RBV. No patient experienced severe anemia (hemoglobin 

level < 8 g/dL) during treatment. In non-haemodialysed 

patients, GFR was not significantly modified during treatment 

(29.6±6.2 mL/min at base line vs. 27.9±6.5 mL/min at the 

end of treatment). No patient had treatment discontinuation 

and antiviral therapy was well tolerated. Conclusion. Our 

results strongly suggest that SOF-based antiviral therapy, with 

reduced dose of SOF, is safe and effective for the treatment of 

HCV patients with ESRD, including haemodialysed, similarly 

to HCV patients without ESRD. Final SVR12 will be presented. 

Disclosures: 

Jérôme Dumortier - Board Membership: Novartis, Astellas, Roche; Consulting: 

Novartis; Grant/Research Support: Novartis, Astellas, Roche, MSD, GSK 

François Bailly - Board Membership: ABBVIE, MSD, BMS, GILEAD; Speaking and 

Teaching: JANSSEN 

Georges-Philippe Pageaux - Advisory Committees or Review Panels: Roche, 

Roche, Roche, Roche; Board Membership: Astellas, Astellas, Astellas, Astellas 


Roche 

François Habersetzer - Advisory Committees or Review Panels: Cytheris; Board 

Membership: Gilead, BMS; Speaking and Teaching: Gilead, Janssen, Roche, 

BMS 




MSD, Abbvie 



The following authors have nothing to disclose: Sylvie Radenne, Marie-Claude 

Gagnieu, Jean Didier Grange, Anne Minello, Nassim Kamar 

1159 

Therapeutic effect of dual oral therapy with Daclatasvir 

and Asunaprevir 

Motoyuki Kohjima 1,2 , Kenta Motomura 1 , Toshimasa Koyanagi 1 , 

Seiya Tada 1 , Takeaki Satoh 1 , Naoki Yamashita 1 , Masaki Yokota 1 , 

Rie Sugimoto 1 , Syoji Nagase 1 , Syusuke Morizono 1 , Nobito Higuchi 

1 , Tsuyoshi Yoshimoto 1 , Shigeki Tashiro 1 , Yuki Tanaka 1 , Kunitaka 

Fukuizumi 1 , Kazuhiro Kotoh 1 , Makoto Nakamuta 1 ; 1 Fukuoka 

Kanzo Treatment research (FKT) group, Fukuoka, Japan; 2 Gastroenterology, 

Clinical Research Center, Kyushu Medical Center, 

National Hospital Organization, Fukuoka, Japan 

Background: The dual oral therapy with the first-in-class NS5A 

replication complex inhibitor Daclatasvir (DCV) and the 

potent NS3 protease inhibitor Asunaprevir (ASV) has recently 

approved for the treatment of chronic hepatitis C genotype 1 

patients. Previous trials for the treatment was shown promising 

results for improving SVR to more than 80% in patients with 

HCV genotype 1b. We conducted a prospective, multicenter 

study to investigate the effectiveness of the dual oral therapy 

with DVC and ASV. Methods: The patients with HCV genotype 

1b have been treated with DCV and ASV since Sep. 2014 

(n=523). The enrolled population was generally older (median 

69 years old) that was consistent with HCV epidemiology in 

Japan and predominantly female (63%). HCV genotype and 

IL-28B polymorphisms were determined by PCR amplification 

and sequencing. HCV resistant associated polymorphisms 

were analyzed by direct sequence of the HCV NS3 and NS5A 

domains. Results: HCV-RNA declined rapidly after the initiation 

of the DCV + ASV treatment, and HCV-RNA was lower than 

detection sensitivity limit in 69% of the patients after 2 weeks 

and 82% of patients achieved RVR. The viral response of HCV 

was not affected by age, sex, liver fibrosis, previous treatment 

response, or IL-28B SNPs. The positive rate for HCV-RNA at 

4 weeks of treatment was significantly higher in patients with 

resistance-associated polymorphism in NS3 or NS5A and RVR 

rate was significantly higher in patients with add-on lipid modulators, 

pitavastain and EPA. Although 19 patients experienced 

viral breakthrough, 8 patients had no resistance-associated 

polymorphism in NS3 or NS5A among the patients with viral


breakthrough. Only 2 cases experienced viral relapse and 

93% of patients achieved SVR 4w. The response rate was 

tend to be higher in patients treated with pitavastain and EPA 

and significantly lower for patients with resistance-associated 

polymorphism (80%, p= 0.002) or treated cases with simeprevir 

(20%, p


ropoietin. Among pts receiving TPV+pegIFN/RBV, 12 (10%) 

were transfused and 1 received erythropoietin due to anemia. 

Conclusions: In MALACHITE-I and MALACHITE-II, 3D+/-RBV 

had more favorable AE profiles than TPV+pegIFN/RBV. With 

3D+/-RBV, anemia was less frequent and less severe, and 

never required transfusion or use of erythropoietin. RBV was 

better tolerated in the context of newer DAAs than of TPV and 

pegIFN. 

Adverse events and hemoglobin decreases in MALACHITE-I and 

MALACHITE-II. 

*P1755μg.h/L after the first dose is predictive of sustained virological 

response (SVR) in HCV-patients treated with peginterferon/RBV*. 

The aim of this study was to test the clinical benefit 

of RBV early dose adjustment based on this threshold (Bayesian 

estimation) in under-exposed naïve genotype 1-infected 

patients. Methods: multicenter, randomized, controlled trial 

with two parallel groups; SC-group: standard of care in 2010- 

2011, i.e. peginterferon-α2a 180μg/week and weight-based 

RBV 1000-1200mg/day during 48 weeks; AD-group: personalized 

increase of RBV dose if D0AUC 0-4H 


Disclosures: 




Pierre Marquet - Advisory Committees or Review Panels: Astellas France, Sandoz 

France, BMS France; Consulting: Chiesi 

The following authors have nothing to disclose: Marianne Maynard-Muet, Sylvie 

Thevenon, Pierre Pradat, Annick Rousseau, Marie-Claude Gagnieu, Sophie 

Alain, Paul Deny, Françoise Lunel-Fabiani, Nicolas Picard, Irène Zublena, Christian 

Trepo 

1163 

Treatment of Hepatitis C Genotype 4 patients with Simeprevir 

and Sofosbuvir: Preliminary Results from a Phase 

IIa, Partially Randomised, Open-label Trial conducted in 

Egypt (OSIRIS) 

Maissa El Raziky 2 , Mohamed Gamil 2 , Radi Hammad 3 , Mohammed 

Saad Hashem 4 , Marwa Khairy 2 , Aisha Elsharkawy 2 , Asmaa 

Gomaa 4 , Sofia Keim 5 , Gino Van Dooren 1 , Robert Ryan 6 , Ralph 

DeMasi 6 , Isabelle Londjon-Domanec 7 , Mohamad Hassany 3 , 

Wahid H. Doss 3 , Imam Waked 4 ; 1 Janssen Infectious Diseases 

BVBA, Beerse, Belgium; 2 Cairo University, Cairo, Egypt; 3 National 

Hepatology and Tropical Medicine Research Institute, Cairo, 

Egypt; 4 National Liver Institute, Menoufiya, Egypt; 5 Janssen-Cilag 

Portugal, Lisbon, Portugal; 6 Janssen Research & Development LLC, 

Titusville, NJ; 7 Janssen-Cilag, Paris, France 

Background and Aims Egypt has the highest prevalence of 

chronic hepatitis C virus (HCV) infection in the world, with 

>90% of patients infected with HCV G4. OSIRIS is a Phase IIa, 

open-label trial evaluating the efficacy and safety of 8 or 12 

weeks’ treatment with once-daily simeprevir (SMV) + sofosbuvir 

(SOF) for HCV genotype 4-infected patients in Egypt. Methods 

Sixty-three treatment-naïve or prior peg-interferon/ribavirin (PR) 

treatment-experienced patients with HCV G4 were treated with 

SMV 150 mg + SOF 400 mg daily, and were assigned to 

one of two groups; Group A (n=40): patients without cirrhosis 

were stratified by treatment history and fibrosis score, and 

randomised 1:1 to receive either 8 or 12 weeks of treatment; 

Group B (n=23): patients with cirrhosis, received 12 weeks 

of treatment. Absence of cirrhosis was documented by liver 

biopsy using METAVIR score. Cirrhosis was documented by 

either liver biopsy or FibroScan score >14 kPa; cirrhotic 

patients had to have platelet count >50 000/mm 3 and serum 

albumin >3 g/dL. The primary endpoint was the proportion 

of patients with HCV RNA 6 x10 6 IU/mL. One 

treatment-naïve patient in the 12-week arm with F1 fibrosis and 

BVL 1.2 x10 6 IU/mL relapsed. Initial safety data showed no 

discontinuations due to adverse events (AE), and no grade 3 

or 4 treatment-related adverse events have been reported. One 

serious AE (Grade 2 pleural effusion and pulmonary hypertension) 

was observed in a 69-year-old cirrhotic female patient 

after 6 weeks of treatment. This was unrelated to treatment and 

resolved in 2 weeks without treatment discontinuation. Conclusions 

SMV+SOF for 12 weeks in patients with HCV G4 is 

generally safe and well tolerated. High SVR4 rates (95–100%) 

were seen with 12 weeks’ treatment independently of prior PR 

response or cirrhosis. Final SVR12 data will be presented. 

Disclosures: 

Maissa El Raziky - Grant/Research Support: Janssen pharmaceuticals 

Mohamed Gamil - Speaking and Teaching: Bristol-Myers Squibb, Gilead Sciences, 

Janssen Pharmaceuticals, Merck Sharp & Dohme, Roche 

Radi Hammad - Grant/Research Support: Gilead Sciences,Inc, Janssen Pharmaceuticals, 

Inc. 

Sofia Keim - Employment: Janssen-Cilag Farmaceutica; Stock Shareholder: Johnson 

& Johnson 

Gino Van Dooren - Stock Shareholder: Johnson and Johnson 

Robert Ryan - Employment: Janssen Pharmaceuticals; Stock Shareholder: Janssen 

Pharmaceutical 

Ralph DeMasi - Employment: Janssen Pharmaceuticals 

Isabelle Londjon-Domanec - Employment: Janssen 


The following authors have nothing to disclose: Mohammed Saad Hashem, 

Marwa Khairy, Aisha Elsharkawy, Asmaa Gomaa, Wahid H. Doss, Imam 

Waked 

1164 

Limited Effectiveness of Sofosbuvir and Ribavirin for 

the Treatment of HCV GT-3 in Patients with Cirrhosis: A 

Real-World GT-3 HCV Treatment Experience 

Shirin Nafisi, Garrett Fante, Ann Moore, Ashley Hepner, Justin A. 

Reynolds, Karen Arendt, Yvette Cummings, Robert Gish, Richard 

A. Manch, Anita Kohli; Hepatology, St. Joseph’s Hospital and 

Medical Center, Scottsdale, AZ 

Recently approved direct-acting antivirals (DAA) regimens have 

improved the outcomes for many chronic hepatitis C (CHC) 

infected patients, particularly with genotypes 1, 2 and 4. Realworld 

non-clinical trial data on patients with CHC genotype 

3 (GT3) and new DAA regimens is lacking. AIM: To evaluate 

the effectiveness of DAA containing regimens for treatment 

of HCV GT3 in a real-world patient population and to compare 

to outcomes with previous use of pegylated interferon 

(PEG) and ribavirin (RBV). METHODS: We performed a retrospective 

analysis of patients with CHC GT3 treated between 

1/2012-6/2015 (n=73) at a community based clinic within 

an academic medical center in Phoenix, AZ or by ECHO telemedicine 

to rural clinic sites. Demographics, CHC treatment 

regimens, liver fibrosis, virologic data, and psychiatric history 

were collected throughout treatment and post-treatment from 

patient charts. RESULTS: Since 2012, 73 GT3 patients started 

treatment for CHC and are included in this analysis. 81% of 

patients were Caucasian, 14% Hispanic, and 1.7% Native 

American. 66% of patients were men, the mean age was 53 

yrs. (range: 21-72 yrs.) and mean HCV viral load 2,015,535 

IU/mL (range: 72-25,000,000 IU/mL). 27% of patients had 

cirrhosis by imaging, Fibrosure or liver biopsy. 55% of patients 

had a psychiatric illness. 35.1% of patients were previously 

treated with PEG containing therapy. 71.2% (n=52) were 

treated with the sofosbuvir (SOF)/RBV for 24 weeks, 1.3% 

(n=1) treated with SOF/RBV for 48 weeks, 5.4% (n=4) were 

treated with SOF/PEG/RBV for 12 weeks, 20.5% (n=15) were 

treated with PEG/RBV for 24 weeks, and 1.3% (n=1) were 

treated with ledipasvir (LDV)/SOF/RBV for 12 weeks. Overall, 

66% percent of patients treated with SOF/RBV for 24-48 

weeks and who have reached the SVR12 timepoint (n=30) 

have achieved SVR12 (pending for 23 patients). Comparing 

patients with or without cirrhosis who have reached SVR12


and were treated with SOF/RBV, 33% of these patients with 

cirrhosis achieved SVR12 as compared to 91% of patients 

without cirrhosis. 100% of patients treated with PEG/SOF/RBV 

who reached the SVR12 timepoint (n=2) have achieved SVR12 

(pending for 2 patients). 80% of patients treated with PEG/RBV 

achieved SVR12. Complete SVR12 results will be presented. 

CONCLUSION: Treatment of CHC GT3 using the PEG sparing 

regimen of SOF/RBV has poor results in cirrhotic patients in a 

real-world clinic setting. Consideration of alternative regimens, 

including SOF/PEG/RBV may be reasonable for these select 

patients or to wait for new treatments. Further development 

of PEG sparing DAA regimens for the treatment CHC GT3 is 

necessary for this difficult to treat population. 

Disclosures: 





The following authors have nothing to disclose: Shirin Nafisi, Garrett Fante, Ann 

Moore, Ashley Hepner, Justin A. Reynolds, Karen Arendt, Yvette Cummings, 

Anita Kohli 

1165 

Efficacy and Safety of Sofosbuvir-based Regimens in 

Asian-Americans With Chronic Hepatitis C Virus (HCV) 

Mono-Infection: A Multi-Center Study in the United 

States 

Calvin Q. Pan 1 , Elsa Ouyang 2 , Myron Tong 3 , Albert Min 4 , Ke-Qin 

Hu 5 , James Park 1 ; 1 Division of Gastroenterology and Hepatology, 

NYU Langone Medical Center, NYU School of Medicine, Flushing, 

NY; 2 Columbia University, New York, NY; 3 Digestive Diseases 

and Liver Center, David Geffen School of Medicine at UCLA, Los 

Angeles, CA; 4 Gasterenterology and Hepatology, Mount Sinai 

Beth Israel, Mount Sinai School of Medicine, New York, NY; 5 Division 

of Gastroenterology and Hepatology, UC Irvine Medical Center, 

Orange, CA 

Background Treatment with sofosbuvir (SOF)-based regimens 

for HCV infection has resulted in sustained virologic response 

(SVR) rates of approximately 90% in pivotal trials, in which 

Asian patients were underrepresented. This study aims to 

assess the efficacy and safety of SOF-based therapy in the 

Asian-American patients. Methods Asian patients with HCV 

genotype 1-6 mono-infection, who received SOF-based therapy 

for 12 or 24 weeks, were retrospectively enrolled from 

multiple centers throughout the United States. The primary endpoint 

was SVR 12. Secondary endpoints were the safety and 

tolerability of SOF-based treatment regimens. Results Among 

the 80 patients enrolled, 45 were treated with SOF+ribavirin 

(RBV), 15 with SOF+simprevir (SIM), 10 with SOF+RBV+peginterferon, 

8 with ledipasvir+SOF and 2 with SIM+SOF+RBV. 

Patient baseline values are shown in Table 1. By week 4, a 

rapid decrease in HCV RNA levels to


patients. Cox analyses showed that SVR was associated with 

reduced occurrence of cirrhosis-related complications (adjusted 

HR 0.19, 95%CI 0.10-0.35). The overall 5-year event rate was 

22.4% (95%CI 18.3-26.5) among those without SVR. With 

95% SVR the NNT to prevent 1 cirrhosis-related complication 

in 5 years was 27, 14, 9 or 7 at a 5-year complication rate of 

5, 10, 15 or 20%, respectively. With 85% SVR this was 30, 

15, 10 or 8, respectively. CONCLUSION The clinical efficacy 

of interferon-free therapy among patients with advanced but 

compensated liver disease is high, especially in case of an 

increased risk of cirrhosis-related complications. Small differences 

in the high SVR rates of these regimens have only limited 

effect on the clinical efficacy 

Disclosures: 


Gilead 

















Novartis 






Bart J. Veldt - Board Membership: GSK, Janssen Therapeutics 






The following authors have nothing to disclose: Giovanna Fattovich, Frank Lammert, 

Wolf P. Hofmann, Donatella Ieluzzi, Bettina E. Hansen 

1167 

Early response and efficacy of dual oral therapy with 

asunaprevir and daclatasvir for elderly patients with 

hepatitis C 

Takashi Honda, Masatoshi Ishigami, Yoji Ishizu, Teiji Kuzuya, 

Kazuhiko Hayashi, Yoshiki Hirooka, Hidemi Goto; Gastroenterology 

and Hepatology, Nagoya University Graduate School of 

Medicine, Nagoya, Japan 

Background and aim: In Japan, patients with hepatitis C virus 

(HCV)-associated liver disease including hepatocellular carcinoma 

(HCC) are getting older and many patients die of such 

disease. The efficacy of dual oral therapy by using asunaprevir 

(NS3 protease inhibitor) and daclatasvir (NS5A replication 

complex inhibitor) for elderly patients with HCV genotype 1b 

has not been clarified. The aim of this study was to evaluate 

the early response and efficacy of dual oral therapy in such 

patients. Methods: Four hundred forty two consecutive chronic 

patients with HCV genotype 1b including compensated cirrhosis 

were treated with dual oral therapy with asunaprevir 

and daclatasvir in our hospital and affiliated hospital. These 

patients were divided into two groups according to age: 

elderly patients (aged ≥ 70 years, n = 240) and non-elderly 

patients (aged < 70 years, n = 202). Clinical characteristics, 

the rapid virological response (RVR) and the sustained virological 

response at post-treatment week 4 (SVR4) obtained 

by intention-to-treat analysis were compared between the two 

groups. Biochemical response and α-fetoprotein were compared 

at the start point and 4weeks later. Results: The ratio of 

female and compensated cirrhosis was significantly higher in 

elderly patients than in non-elderly patients (p = 0.006, 0.003, 

respectively). ALT, gGTP and hemoglobin level was significantly 

lower in elderly patients than that in non-elderly patients 

(P =0.0001, 0.010, 0.0001, respectively). However, other 

clinical characteristics of patients were not significantly different 

between two groups. The RVR and SVR4 rates didn’t differ 

significantly between elderly patients and non-elderly patients 

(RVR: 94.3 % vs 97.0 %, SVR4: 84.6 % vs 92.1 %). According 

to multivariate analysis, only RVR proved to be significantly 

associated with SVR4 while patient age did not affect SVR4. 

Multivariate analysis also indicated there is no significant factor 

associated with SVR4 in elderly patients. HCVRNA, ALT, 

γGTP and α-fetoprotein was rapidly and significantly reduced 

from start point to 4 weeks later in both elderly patients and 

non-elderly patients. Conclusions: Treatment of chronic hepatitis 

C including compensated cirrhosis with dual oral therapy was 

comparably effective between elderly patients and non-elderly 

patients. This dual oral therapy rapidly reduced ALT, γGTP and 

α-fetoprotein and may reduce incidence of HCC even in elderly 

patients. 

Disclosures: 



The following authors have nothing to disclose: Takashi Honda, Masatoshi Ishigami, 

Yoji Ishizu, Teiji Kuzuya, Kazuhiko Hayashi, Yoshiki Hirooka


1168 

Resistance Analyses of Phase 3 Studies in Korean and 

Taiwanese Patients with Chronic Hepatitis C Receiving 

Sofosbuvir or Ledipasvir/Sofosbuvir Containing Regimens 

Kwang-Hyub Han 2 , Brian Doehle 1 , Hadas Dvory-Sobol 1 , Evguenia 

S. Svarovskaia 1 , Ramakrishna K. Chodavarapu 1 , Jenny C. Yang 1 , 

Steven J. Knox 1 , Michael D. Miller 1 , Hongmei Mo 1 , Wan-Long 

Chuang 3 ; 1 Gilead Sciences, Foster City, CA; 2 Severance Hospital, 

Yonsei University Health System, Korea (the Republic of); 3 Kaohsiung 

Medical University Hospital, Kaohsiung City, Taiwan 

Background: The prevalence of chronic HCV infection varies 

globally, with approximately 1% and 4% of the population 

infected in Korea and Taiwan, respectively. Sofosbuvir (SOF) 

containing regimens can provide safe and highly effective all 

oral treatment regimens for patients infected with genotype 

(GT) 1 or 2 HCV. In this study we evaluated the impact of preexisting 

resistant-associated variants (RAVs) on treatment outcome 

and emergence of RAVs at relapse in patients retreated 

with SOF+RBV or ledipasvir (LDV)/SOF for 12 weeks in 

Korea and Taiwan. Methods: LDV/SOF (n=178) or SOF+RBV 

(n=216) was administered for 12 weeks to Korean or Taiwanese 

patients chronically infected with GT1 or 2 HCV. NS5A 

and/or NS5B deep sequencing analysis (cut-off of 1%) was 

performed for patients at baseline according to the assigned 

treatment regimen and at the time of virologic failure in patients 

failing to achieve SVR12. Data were compared to reference 

sequences and patient baseline sequence for patients failing 

to achieve SVR. Results: Virologic failure rates were low 

in Korean and Taiwanese patients; 1.1% (2/178) in LDV/ 

SOF (GT1) recipients and 0.9% (2/216) in those receiving 

SOF+RBV (GT2). HCV GT1: Of the 178 patients with GT1 

HCV infection, 5 patients were incorrectly genotyped by LiPA 

and corrected to GT6 by sequence analysis; all 5 achieved 

SVR. Thirty-eight of 172 patients (22%) with available NS5A 

sequence data were observed to have baseline NS5A RAVs 

with 37/38 (97%) achieving SVR. The Y93H (n=30) and 

L31F/I/M/V (n=8) NS5A RAVs as single variants were the 

most commonly observed in both populations. Four of the 170 

patients with available sequence data (2%) were observed to 

have the NS5B RAV L159F at baseline and all achieved SVR. 

In the two GT1 patients that relapsed post treatment, one was 

observed to have NS5A RAVs at baseline and relapse, and the 

other had treatment emergent NS5A RAVs. No NS5B RAVs 

were observed in these two patients. HCV GT2: Six of 184 

patients (3%) with available baseline NS5B sequencing data 

were observed to have M289I/L variants, and all achieved 

SVR when treated with SOF+RBV. No other baseline NS5B 

RAVs were observed. Two GT2 patients experienced virologic 

failure, and no baseline or treatment emergent NS5B RAVs 

were observed. Conclusions: Low rates of virologic failure were 

observed in Korean and Taiwanese patients and the presence 

of NS5A and NS5B RAVs did not affect the treatment outcome 

to LDV/SOF and SOF+RBV in patients with GT1 and GT2 

infection, respectively. No S282T or other SOF treatment-associated 

variants have been detected in the patients that experienced 

virologic failure during treatment. 

Disclosures: 



Sciences 

Evguenia S. Svarovskaia - Employment: Gilead Sciences Inc.; Stock Shareholder: 

Gilead Sciences Inc. 

Ramakrishna K. Chodavarapu - Employment: Gilead Sciences, Inc 








The following authors have nothing to disclose: Kwang-Hyub Han 

1169 

Sustained Virologic Response (SVR) of Liver Transplant 

Recipients (OLT) with Advanced Hepatitis C Recurrence 

(HCVR), All Genotype 1, after Combination Antiviral 

Therapy (Rx) with either Telaprevir(TVR) or Sofosbuvir/ 

Simeprevir (SOF/SIM): A Single Center Experience 

Carlos Fasola, Parvez S. Mantry, Jeffrey S. Weinstein, Hector E. 

Nazario, Adil Habib, Maisha Barnes, Alejandro Mejia, Edward 

A. Dominguez, Richard Dickerman, Stephen Cheng; Liver Institute, 

Methodist Dallas Medical Center, Dallas, TX 

Aim: Liver allografts of viremic HCV-OLT have a higher risk of 

failure. Timely intervention is necessary. However, long-term 

outcome reports, especially using newer Rx, is lacking. Methods: 

in a 4-year period, we compared 2 groups of OLT-HCVR 

Rx: A) TVR (n=19) + Pegylated interferons (P-IFN) + Ribavirin 

(RBV) for 12 weeks, followed by P-IFN+RBV for 12-36 weeks 

or, B) SOF/SIM (n=28) Rx for 12-24 weeks. Rx criteria: OLT 

graft dysfunction and/or HCVR histology (stage 0-4, Batts&Ludwig). 

Immunosuppression based on tacrolimus or cyclosporine 

+ mycophenolic acid + low-dose prednisone (limited time). 

Demographics, laboratory tests, including HCV-RNA levels, 

graft and OLT survivals were analyzed. Minimal follow: 24 

weeks post end of Rx. Attention focused on rapid virologic 

response (RVR: aviremic in < 4 weeks), early virologic response 

(EVR: aviremic in < 8 weeks) and SVR (aviremic for > 24 weeks 

post Rx). Statistics: t-tests, Chi2-tests and Kaplan-Meier. Results: 

The majority of the patients (14/19 and 21/28, 74-75%) were 

non-responder or intolerant to previous Rx. Only a quarter of 

patients were naive to HCV Rx on each group. No demographic 

or laboratory differences were found, except for a 

similar incidence of anemia that required epoetin Rx. Liver tests 

remained normal in 26/28 (93%) of the SOF/SIM group. The 

pre-Rx advanced stage was significantly higher in the SOF/ 

SIM group (table). No differences were observed regarding 

patient and allograft survivals. Three patients (16%) died due 

to allograft failure in group A. In group B, 2 (7%) died, one 

of them of trauma with stable LFT’s. Three patient relapsed in 

the TVR group. No differences, between groups, were found 

in the rate of HCV-RNA levels decrease log during the first 8 

weeks, however there was a significant difference in the SVR 

rates favoring the SOF/SIM combination (table). Conclusions: 

Newer anti-HCV therapies have significantly improved the SVR 

and outcome of OLT recipients with HCVR. The benefit of SOF/ 

SIM combination therapy has shown, in our experience, to be 

an important addition to the effort of rescueing liver allograft 

function post OLT in HCVR recipients. 

Characteristics and Outcome of Liver Transplant Recipients with 

Hepatitis C Recurrence after Antiviral Therapy 

P value < 0.05: 1 vs. 1 and 2 vs. 2


Disclosures: 





Salix 

Jeffrey S. Weinstein - Speaking and Teaching: Merck 

Hector E. Nazario - Advisory Committees or Review Panels: Gilead, Janssen; 

Speaking and Teaching: Gilead, Merck, Abbvie, Janssen 

Edward A. Dominguez - Advisory Committees or Review Panels: Gilead, Pfizer; 

Grant/Research Support: Cubist; Speaking and Teaching: Amgen, Astelleas 

The following authors have nothing to disclose: Carlos Fasola, Adil Habib, Maisha 

Barnes, Alejandro Mejia, Richard Dickerman, Stephen Cheng 

1170 

Treatment of Chronic HCV Genotype 1 (G1) Infection 

with Boceprevir: Frequency, Severity, Predictability and 

Management of Anemia in German Real-Life 

Gerlinde Teuber 1 , Peter Buggisch 2 , Hanns F. Loehr 3 , Hermann 

Steffens 4 , Michael R. R. Kraus 5 , Peter R. Geyer 6 , Bernd Weber 7 , 

Thomas Witthoeft 8 , Uwe Naumann 9 , Elmar Zehnter 10 , Dagmar 

Hartmann 11 , Bernd Dreher 11 , Manfred Bilzer 11 ; 1 Gastroenterological 

Practice, Frankfurt, Germany; 2 IFI Liver Center Hamburg, 

Hamburg, Germany; 3 Gastroenterological Practice, Wiesbaden, 

Germany; 4 Practice of Internal Medicine, Berlin, Germany; 

5 Department II, Klinikum Burghausen, Burghausen, Germany; 

6 Gastroenterological Practice, Fulda, Germany; 7 Competence 

Center Addiction, Kassel, Germany; 8 Gastroenterological Practice, 

Stade, Germany; 9 Center of Medicine, Berlin, Germany; 10 Gastroenterological 

Practice, Dortmund, Germany; 11 MSD Pharma 

GmbH, Haar, Germany 

Background: Anemia is a frequent complication in patients 

(pts) undergoing triple therapy with the HCV protease inhibitor 

boceprevir (BOC). Data regarding the frequency, severity, 

predictability and management of anemia caused by triple 

therapy in real-life are still scarce and were evaluated in the 

present interim analysis. Methods: From April 2012 until January 

2014, 536 pts with HCV G1 infection were recruited in 

the ongoing NOVUS observational study by 97 practices and 

hospitals in Germany. Pts were treated with pegylated interferons 

(PegIFN) and ribavirin (RBV) together with BOC for 24 to 

44 weeks after a 4 weeks lead-in period with PegIFN/RBV. The 

present interim analysis was restricted to 265 treatment-naïve 

and 132 previously treated pts with documented hemoglobin 

(Hb) levels at baseline and during therapy. Results: During 

BOC triple therapy anemia (Hb


past regimens in achieving SVR in this difficult to treat population. 

Table 1. Main demographic, baseline clinical, virological and 

laboratory (values expressed as mean ± standard deviation) characteristics 

of HCV-MC patients 

or ombitasvir/paritaprevir/ritonavir/dasabuvir for GT1 and 

sofosbuvir/ribavirin for GT 2/3. Data was included from 13 

additional patients who are still awaiting medication approval 

for a total of 54 patients. Among these 54 patients, HCV 

medications were approved in 84% of patients with private 

insurance, 71% of Medicare and 50% of Medicaid patients 

(p=0.11). There was no difference in approval rate based 

on age, race, gender, genotype, treatment regimen or presence 

of cirrhosis. The median time spent per patient was 92.5 

minutes (IQR 80.0-105.0). The median cost spent per patient 

was $60.54 (IQR 53.00-72.90). There was no difference in 

time spent acquiring HCV medications or cost per patient by 

genotype, insurance status, treatment regimen or presence of 

cirrhosis. Conclusion: In a Hepatology practice with experienced 

staff using a specialty pharmacy to improve efficiency, 

the median provider time needed to obtain DAAs was 90 minutes 

per patient. This relatively labor intensive process will need 

to be streamlined to encourage more clinicians to engage in 

HCV therapy for the large number of potential candidates in 

the United States. 

Disclosures: 

The following authors have nothing to disclose: Veronica Loy, Tamara Benyashvili, 

Megan OMahony, Douglas Pavkov, William Adams, Robin Todus, Michelle 

Watts, Natasha Von Roenn, Scott Cotler 

HCV:hepatitis c virus 

MC: mixed cryoglobulinemia 

Disclosures: 


The following authors have nothing to disclose: Rasham Mittal, Zoe Bider-Canfield, 

Amandeep K. Sahota 

1172 

The Time and Cost Investment Required to Obtain and 

Initiate Direct Acting Antiviral Therapy 

Veronica Loy, Tamara Benyashvili, Megan OMahony, Douglas 

Pavkov, William Adams, Robin Todus, Michelle Watts, Natasha 

Von Roenn, Scott Cotler; Hepatology, Loyola Medical Center, Chicago, 

IL 

The biggest barrier to direct-acting antiviral agent (DAA) therapy 

for hepatitis C (HCV) is cost. Initiating treatment with DAAs 

can be time intensive for the provider, including obtaining 

prior authorization. The purpose of this study was to assess the 

amount of unbillable time and cost needed to obtain DAAs for 

HCV. Methods: We prospectively enrolled patients for whom 

DAA therapy was planned from 9/30/2014- 3/19/2015. 

Providers recorded the amount of time spent in minutes to 

obtain medication and to initiate HCV therapy for each patient. 

The position of each provider (APN, RN, LPN) was noted. Data 

included age, gender, race, HCV genotype, type of insurance 

carrier, and presence of cirrhosis. Patients who subsequently 

defered therapy and those with incomplete time data were 

excluded from data analysis. In our practice, 2 LPNs and 6 

RNs collect patient information and use a specialty pharmacy 

to obtain approval for DAAs. An APN reviews potential medication 

interactions and conducts a patient education session 

by phone. Time spent was converted to cost per patient based 

on average hourly medical wages in Illinois by Salary.com. 

Administrative time and cost per patient was compared by 

insurance carrier, HCV genotype, treatment regimen and presence 

of cirrhosis. Results: 79 patients consented to participate 

in the study. 25 were excluded due to incomplete time data 

(n=17) or subsequent deferral of therapy (n=8). 41 patients 

were started on therapy consisting of sofosbuvir/ledipasvir 

1173 

Health resource usage in peri-transplant patients during 

12 weeks of therapy with sofosbuvir (SOF), with or 

without ribavirin (RBV) and an NS5A inhibitor 

Suman Verma 1 , Graham R. Foster 2 , William Irving 3 , John McLauchlan 

5 , Michelle C. Cheung 2 , Benjamin E. Hudson 4 , Ivana Carey 1 , 

Kosh Agarwal 1 ; 1 Institute of liver studies, Kings College Hospital, 

London, United Kingdom; 2 The Blizard Institute, Queen Marys University 

of London, London, United Kingdom; 3 Virology, University 

of Nottingham, Nottingham, United Kingdom; 4 University of 

Nottingham, Nottingham, United Kingdom; 5 MRC University of 

Glasgow, London, United Kingdom 

Introduction Direct acting antiviral (DAA) therapy for HCV 

decompensated disease is a reality. However, health economic 

benefit in this advanced peritransplant population remains 

unclear. This study aims to assess healthcare resource utilization 

and quality returns from a clinical and cost perspective 

during 12 weeks (wks) of DAA therapy in this challenging population. 

Methods The UK NHS England Early Access Program 

(EAP) mandated antiviral therapy to HCV positive patients with 

advanced liver disease via 17 centres. Twelve weeks of therapy 

with SOF and an NS5A inhibitor (Ledipasvir or Daclatasvir), 

with or without RBV was administered to a cohort of 

~500 decompensated cirrhostics, with 179 peritransplant (PT). 

PT was defined as meeting the NHSBT criteria for liver transplantation 

(LT) (n=56); on the national LT waiting list (n=50); 

or a previous LT with advanced HCV recurrence in graft and 

portal hypertension (n=45). Choice of therapy was clinician 

discretion. Data collection was prospective during the 12 wks 

of therapy via HCV Research UK on use of blood products; 

patient phone calls made to healthcare professionals including 

specialist hepatitis nurses (SHN) and consultants; unscheduled 

clinic appointments; and elective and emergency admissions 

into hospital and intensive care (ITU). Results Of the179 PT 

patients enrolled, 151 had completed 12 wks of therapy at 

the time of abstract submission. During this, 148 phone calls 

were made to SHN; 5 to hepatology consultants and 42 to 

other healthcare professionals (OHP) including GPs and clinical 

pharmacists, with a total duration of 993, 25, 280 minutes 

respectively. A total of 367 unscheduled clinic appointments


were required during the treatment period with 178 delivered 

by consultants, 112 by SHN and 77 by OHP. Support for 

anemia with erythropoietin, cosmofer or blood transfusion was 

required equally across the PT cohorts (previous LT 7, fulfilling 

NHSBT criteria 5, listed for LT 7). In addition there were 

133 hospital admissions with 25% electively for ascites drainage, 

and as an emergency 5% for variceal bleed and 9% for 

encephalopathy. Furthermore, 21 patients underwent LT, with 

complete inpatient stay data available for 13. This demonstrated 

an average stay of 18.07 days, with 2.85 days on ITU 

and 1.53 days ventilated. Further data will be presented at 

AASLD specifically addressing financial costs. Conclusion This 

preliminary analysis of health resource usage by PT patients 

during SOF and NS5A inhibitor +/- RBV demonstrates that this 

challenging advanced disease population still utilize significant 

healthcare resource despite better tolerability of the new DAA 

therapies. 

Disclosures: 







William Irving - Advisory Committees or Review Panels: Novartis, MSD, Janssen 

Cilag, Bristol Myers Squibb; Grant/Research Support: GSK, Pfizer, Janssen 

Cilag, Gilead Sciences; Speaking and Teaching: Janssen Cilag, Roche 


BMS 

Kosh Agarwal - Advisory Committees or Review Panels: Gilead, Novartis, 

Abbott; Grant/Research Support: Roche, MSD; Speaking and Teaching: BMS, 

Astellas, Janssen 

The following authors have nothing to disclose: Suman Verma, John McLauchlan, 

Michelle C. Cheung, Benjamin E. Hudson 

1174 

Outcomes of ribavirin free regimens for treatment of 

recurrent hepatitis C (genotype1) infection post liver 

transplant 

Mohamed G. Shoreibah 1 , DeAnn Jones 2 , Sarah Baggett 2 , Brendan 

M. McGuire 1 , Omar Massoud 1 ; 1 GI and Hepatology, University 

of Alabama at Birmingham, Birmingham, AL; 2 Pharmacy, 

University of Alabama at Birmingham, Birmingham, AL 

Background. The development of direct-acting antivirals in 

the treatment of Hepatitis C Virus (HCV) is a rapidly evolving 

field. Data on the use of the newer agent in the treatment 

of recurrent HCV infection post liver transplant (LT) is scarce. 

Methods. We performed a retrospective analysis comparing 

two groups treated with RIBA free regimens for recurrent HCV 

infection, genotype 1, post LT: Group A, treated with SIM/ 

SOF and Group B, treated with LDV/SOF (without RIBA). 

Patients received treatment between October 2014 and April 

2015. Patients were identified through hepatology department 

records. Data collection included genotype, prior HCV therapies, 

cirrhosis, and virologic response at 4 and 12 weeks on 

treatment. A one sided t-test was used to determine statistical 

significance. Results. A total of 78 patients met inclusion criteria 

with mean time from LT of 61 months. Descriptive statistics 

were: Group A (N:36, mean age: 58 years, 69% male, 58% 

treatment experienced, 73% GT1a, 11% cirrhotics, 1 patient 

with fibrosing cholestatic HCV) and Group B (N:42, mean 

age: 60 years, 69% male, 55% treatment experienced, 77% 

GT1a, 7% SOF failure, 10% cirrhotics, ). Group A patients 

were treated for 12 weeks compared to planned 12 weeks in 

76% and 24 weeks in 24% of patients in Group B. In group A, 

100% completed treatment. End of treatment response (EOTR) 

was 94% and available sustained virologic response (SVR12) 

rate was 91%. One patient with fibrosing cholestatic HCV 

achieved SVR12. All three patients who experienced relapse in 

group A are currently enrolled in Group B with a planned 24 

week treatment duration. A total of 50% of patients in Group B 

have completed treatment. Available EOTR is 100 (N:20) and 

available SVR12 is 100% (N:4). A total of 22% of patients 

had renal insufficiency with a mean glomerular filtration rate 

of 45 ml/min at the start of treatment. The mean albumin was 

3.64 gm/dL at the start of treatment and 3.9 gm/dL at SVR12 

(P=0.013), which suggests improved synthetic liver function. 

No discontinuation of treatment due to adverse events occurred 

in either group. Data collection is ongoing. Conclusion. The 

combination of SIM/SOF showed a high efficacy of 91% in the 

treatment of recurrent HCV post LT with excellent tolerability. 

The use of LDV/SOF in this group of patients (without RIBA) is 

being evaluated in our institution and appears to have excellent 

virologic response and tolerability. Additional SVR12 data will 

be reported at the time of the meeting. 

Disclosures: 

Mohamed G. Shoreibah - Advisory Committees or Review Panels: Gilead 

Brendan M. McGuire - Grant/Research Support: bayer healthcare, salix 

The following authors have nothing to disclose: DeAnn Jones, Sarah Baggett, 

Omar Massoud 

1175 

Predictors of Treatment Adherence and Discontinuation 

in Department of Defense (DoD) Health Care Beneficiaries 

Treated for Chronic Hepatitis C 2004-2013 

Dana Y. Teltsch 2 , David R. Walker 1 , Beth L. Nordstrom 2 , Kathy 

Fraeman 2 , Karl Kronmann 3 , Kristina St Clair 3 ; 1 HEOR, Abbvie, 

North Chicago, IL; 2 Retrospective Observational Studies, Evidera, 

Lexington, MA; 3 Naval Medical Center Portsmouth, Portsmouth, 

VA 

Background and objective: New treatments for chronic hepatitis 

C virus (HCV) promise excellent sustained virologic response 

and cure rate. Treatment effectiveness, however, depends on 

regimen compliance. Our goal was to identify predictors of 

adherence and discontinuation in a veteran, dependent and 

active duty military population receiving chronic HCV treatment. 

Methods: We conducted a retrospective cohort study 

of chronic HCV patients covered by the DoD health system 

who initiated HCV treatments during 2004-2013. The defined 

regimens included: peg-interferon (peg)+ribavirin (rib) alone 

or in combination with boceprevir (boc)/telaprevir (tel)/simeprevir 

(sim); or sofosbuvir (sof) in combination with rib/sim/ 

rib+sim/peg+rib; all regimens were studied for 12 weeks after 

initiation. The time to regimen discontinuation was calculated 

allowing up to a 60 (peg and rib) or 15 (other drugs) day gap, 

and predictors of time to discontinuation identified using multivariate 

Cox models. Adherence was measured by proportion 

of days covered (PDC), with PDC > 80% considered adherent. 

Multivariate logistic regression models sought predictors of 

adherence. Potential predictors included patient demographics, 

liver disease status, comorbidities, and baseline resource 

utilization. Results: 6,196 patients initiated treatment with a 

regimen of interest (4,756 peg+rib; 212 peg+rib+boc; 631 

peg+rib+tel; 238 sof+sim; 24 sof+sim+rib; 178 sof+rib+peg; 

157 sof+rib; 0 sim+peg+rib ). Mean age was 49.9 years, and 

47.3% were female. 71.9% of patients had non-cirrhotic HCV; 

26.5% compensated or decompensated cirrhosis; 1.6% were 

post liver transplant. There was a small but consistent association 

between increased time from diagnosis in years and 

both earlier treatment discontinuation (hazard ratio (HR) 1.03; 

95% confidence interval (CI) 1.00-1.06) and lower adherence 

(odds ratio (OR) 0.96; (0.93-0.99)). Predictors of discontinua-


tion included age >60 years at index date (1.18; 1.00-1.38, 

compared to age 51-60); chronic kidney disease (1.51; 1.13- 

2.02), diabetes (1.28; 1.09-1.50), HIV status (1.86; 1.24- 

2.79), and psychiatric disorders (1.14; 1.02-1.28). Predictors 

of adherence included decompensated cirrhosis (0.75; 0.61- 

0.94 compared to non-cirrhotic HCV); chronic kidney disease 

(0.70; 0.50-0.99); diabetes (0.80; 0.68-0.95); and HIV status 

(0.59; 0.36-0.96). Conclusions: Later treatment time from diagnosis, 

patient comorbidities, and older age were associated 

with poor adherence and with discontinuation prior to the recommended 

regimen duration. Results suggest that delaying 

treatment for many years after HCV diagnosis may adversely 

impact treatment compliance. 

Disclosures: 

Dana Y. Teltsch - Consulting: AbbVie; Employment: Evidera 


Healthcare 

The following authors have nothing to disclose: Beth L. Nordstrom, Kathy Fraeman, 

Karl Kronmann, Kristina St Clair 

study suggests as a result of a good tolerability profile, monitoring 

and AE related costs are minimal in LDV/SOF regimens. 

This study also suggests that, when the 8w regimen is used, 

the cost per SVR is significantly lower in naïve and NC when 

compared to TE and cirrhotic patients, indicating an economic 

benefit of selecting high effectiveness and well tolerated first 

line therapies and early treatment. 

Disclosures: 



Karsten Wursthorn - Grant/Research Support: BMS 

Albrecht Stoehr - Board Membership: MSD, B√∂hringer Ingelheim; Speaking 

and Teaching: Janssen, MSD, Gilead, Abbvie, B√∂hringer Ingelheim, BMS, VIIV 

Aline Gauthier - Consulting: Gilead 

Petar K. Atanasov - Consulting: Gilead 





1176 

Real-world effectiveness and cost per SVR of ledipasvir/ 

sofosbuvir chronic hepatitis C treatment 

Peter Buggisch 1 , Karsten Wursthorn 1 , Albrecht Stoehr 1 , Aline 

Gauthier 2 , Petar K. Atanasov 2 , Joerg Petersen 1 ; 1 Asklepios Klinik 

St. Georg Haus L, IFI Institut für Interdisziplinäre Medizin, Hamburg, 

Germany; 2 Amaris Consulting, London, United Kingdom 

Background and Aims: Ledipasvir/Sofosbuvir (LDV/SOF) single 

tablet regimen (STR) is approved in Europe and the US for 

the treatment of chronic hepatitis C (CHC) patients. With the 

emergence of novel, highly effective, and safe therapies and 

the expected demand for them, the need for optimal resource 

allocation is high. The cost per sustained viral response (SVR) 

is a measure which provides insights into the amount spent for 

the achievement of success in CHC therapy. This study aims 

to assess the safety, effectiveness, and the cost per SVR associated 

with LDV/SOF therapy in clinical practice in Germany. 

Methods: The first CHC patients treated with LDV/SOF in a 

single centre (and for whom SVR after 12 weeks of follow-up 

(SVR12) will be available in November 2015) were included 

in the analysis. Baseline characteristics, prior treatment history, 

safety, effectiveness and cost per SVR were investigated 

using descriptive statistics. Results: 115 patients met the inclusion 

criteria for this analysis. 8w (51.3%), 12w (44.3%) or 

24w (4.4%) treatment with LDV/SOF was initiated between 

21/11/2014 - 03/03/2015. 21% of patients had ribavirin 

(RBV) added to the STR (79% of which F4). The mean (SD) age 

was 52 (12.2) years, 60% were male, 89.6% had at least one 

comorbidity. Genotype (GT) distribution: 57%, 32%, 6% and 

4% for 1a, 1b, 3 and 4, respectively. METAVIR stage distribution: 

34%, 19%, 13%, 11% and 23% for F0, F1, F2, F3 and 

F4, respectively. Median (IQR) HCV RNA at baseline was 0.97 

(0.28-1.90) million IU/ml. 6% (2%) of patients were HIV (HBV) 

co-infected. In patients with available outcome data, the SVR4 

was 99% (n=92/93) and SVR12 was 100% (n=13/13). One 

treatment experienced (TE) F4 patient on 12w STR+RBV did not 

achieve SVR4. 6.9% (n=8) experienced grade 3 or 4 adverse 

events (AE) and 5.2% (n=6) were assessed as treatment-related. 

No AE lead to discontinuation. 1.0% of costs were 

non-therapy costs. The median cost per SVR4 was €53,025 (PP 

like approach). 73% of naïve patients and 66% of non-cirrhotic 

(NC) were on 8w duration; median cost per SVR4 was 59% 

lower in NC (€46,775) than in F4 patients and 58% lower in 

naïve (€46,272) versus TE patients. SVR12 and cost per SVR12 

will be available at the time of presentation. Conclusion: This 

1177 

Hope and Hopelessness in HCV patients in the era of 

DAA therapy 

Sophie K. Afdhal 1 , Michael Penn 1 , Zobair M. Younossi 2 , Michael 

P. Curry 3 ; 1 Psychology, Franklin & Marshall College, Charlestown, 

MA; 2 Inova Fairfax Hospital, Falls Church, VA; 3 Beth Israel Deaconess 


Hope and hopelessness are important patient responses to 

chronic illness and can influence recovery from disease and 

effect patient reported outcomes (PRO’s). In the era of Direct 

Acting Antiviral (DAA) therapy with high SVR rates greater 

than 90% and good tolerability we hypothesized that treatment 

expectations might alter the hope of patients with HCV. AIMS: 

To evaluate hope and hopelessness in a HCV population and 

to correlate it with disease variables and PRO’s such as fatigue, 

depression and QOL. PATIENTS and METHODS: Consecutive 

adult patients with chronic HCV were enrolled from the 

Liver Center at BIDMC and underwent education about the 

new DAA therapies and then completed the Beck Hopelessness 

survey (BHS), Beck Depression index (BDI), fatigue questionnaire 

(FACIT-F) and the SF-36 health survey. Demographic, 

social, educational and disease state data was collected. Data 

was analyzed by Chi square for comparison between groups, 

regression analysis and ANOVA. RESULTS: 158 patients were 

enrolled over June to August 2014 and were predominantly 

male (63%), Caucasian (75%) with over 56% having at least 

some college level education. 52% of patients believed that 

HCV was responsible for causing or worsening health related 

issues, 40% had a history of depression, 30% had a history of 

chronic pain, 17% had PTSD, 21% had current drug use and 

35% current alcohol use. Mode of acquisition of HCV was drug 

use in 38%. RESULTS: Mean hopelessness score (range 0 – 20) 

was 4.25 + 4.5 and 25 patients had severe hopelessness with 

a score > 10. BDI mean score 12.6 + 10.6 and 39 patients 

had moderate to severe depression. Patients with a history of 

depression or active depression were strongly correlated with 

hopelessness; P < 0.0001. Patients with PTSD were more likely 

to suffer from hopelessness (p


related to coexisting factors such as depression, PTSD and disease 

beliefs. The effect of successful treatment on hopelessness 

in this cohort is being evaluated. 

Disclosures: 






The following authors have nothing to disclose: Sophie K. Afdhal, Michael Penn 

1178 

Sofosbuvir is well tolerated and effective in chronic hepatitis 

C patients with advanced renal disease and/or on 

hemodialysis 

Geoffrey Kitzman 1 , Eric Davis 1 , Jesus Monico 2 , Evelyn Arendale 1 , 

Brian B. Borg 1 ; 1 Digestive Disease, University of Mississippi Medical 

Center, Jackson, MS; 2 Pathology, University of Mississippi 

Medical Center, Jackson, MS 

Background: Sofosbuvir (Sof) is a nucleotide analog inhibitor 

of HCV NS5B polymerase. It is used in combination therapies 

with ribavirin or other DAAs. The safety and efficacy of sofosbuvir 

in patients with advanced renal disease (GFR


could potentially select F0–2 treatment-naïve G4 patients suitable 

to shorten SMV+PR therapy to 12 weeks. 

Disclosures: 

Tarik Asselah - Consulting: Jannsen, AbbVie, Achillion, BMS, Gilead, Merck, 

Roche; Speaking and Teaching: Jannsen, AbbVie, Achillion, BMS, Gilead, 

Merck, Roche 



















Faisal M. Sanai - Advisory Committees or Review Panels: Merck Sharpe Dohme, 

Bristol Myers Squibb, Janssen Pharmaceuticals, Gilead Sciences; Grant/ 

Research Support: Roche Pharmaceuticals, Bristol Myers Squibb; Speaking and 

Teaching: Roche Pharmaceuticals, Janssen Pharmaceuticals, Gilead Sciences, 

Bayer Schering 

Robert Ryan - Employment: Janssen Pharmaceuticals; Stock Shareholder: Janssen 

Pharmaceutical 

Oliver Lenz - Employment: Janssen; Stock Shareholder: Janssen (J&J) 

Gino Van Dooren - Stock Shareholder: Johnson and Johnson 

Isabelle Londjon-Domanec - Employment: Janssen 

Catherine Nalpas - Employment: Janssen pharmaceuticals 

Michael Schlag - Employment: Janssen Cilag Pharma; Stock Shareholder: Johnson 

& Johnson 



Janssen, BMS 

The following authors have nothing to disclose: Antonio Craxi 

1180 

Rapid drop in serum glucose and hypoglycemia in 

chronic hepatitis C patients with diabetes during oral 

HCV therapy 

Laura M. Benítez-Gutiérrez 1 , Vincent Soriano 2 , Isabella Esposito 1 , 

Pablo Barreiro 2 , Ana Maria Duca 1 , Silvia Requena 1 , Isolina 

Baños 1 , Ana Treviño 1 , Valentin Cuervas-Mons 1 , Carmen de Mendoza 

1 ; 1 Internal Medicine, Hospital Universitario Puerta de Hierro 

Majadahonda, Madrid, Spain; 2 Infectious Disease Unit,, La Paz 

University Hospital & IdiPAZ, Madrid, Spain 

Introduction: Chronic HCV infection is associated with metabolism 

abnormalities, including insulin resistance. Up to 15-25% 

of chronic hepatitis C patients will develop diabetes lifelong. 

New DAA allow HCV suppression within a very short-time 

frame. Little is known on the metabolic effects seen in chronic 

hepatitis C patients on DAA. Methods: All chronic hepatitis C 

patients that received sofosbuvir (SOF)-based regimens during 

at least 4 weeks at two reference hepatitis clinics in Madrid 

were retrospectively examined. Results: A total of 61 chronic 

hepatitis C patients begun SOF-based regimens and had 

weeks 0 and 4 controls. Regimens were as follows: SOF+simeprevir 

(16), SOF+daclatasvir (13), SOF+ledipasvir (27) and 

SOF+ribavirin (5). At 4 weeks 63% had undetectable viremia 

and the rest had


and 8( 22.9) received everolimus. 31(24 genotype 1a, 9 genotype 

1b) patients completed a 24 week course of therapy, 

the remaining 4 patients completed an 8 (genotype 1a),12 

(genotype 1a),15 (genotype 1b) and 16 (genotype 1a) weeks 

of therapy individually. Side effects were reported in six of the 

patients and they included fatigue, somnolence, headache, 

anemia, photosensitivity and weight gain. In genotype 1a, 

4(16%) patients had RVR, 35 (100%) patients had ETR and 

SVR at 4 and 12 weeks. In genotype 1b, 2 (20%) patients had 

RVR, 8(80%) patients had ETR and SVR at 4 and 12 weeks. 

One of the non responders discontinued treatment before completion 

due to decompensation. Overall response rate for genotype 

1a and 1b was 94.3%. No patient required adjustment in 

IS and no episodes of rejection were documented during treatment. 

Conclusion: In patients with recurrent genotype 1 (a or b) 

hepatitis C after LT, the combination of once daily sofosbuvir 

and simeprevir is generally well tolerated with very high overall 

response rates. Discontinuation of treatment only occurred in 

two patients due to decompensation of cirrhosis. There were 

no interactions that required adjustments in IS. 

Disclosures: 

Nikolaos Pyrsopoulos - Advisory Committees or Review Panels: GILEAD, BMS, 

ABBVIE, VITAL THERAPIES, Quest; Grant/Research Support: ABBVIE 

The following authors have nothing to disclose: Nneoma O. Okoronkwo, George 

Protopopas, Kiran V. Rao, Vivek A. Lingiah, Maliha Ahmad, Lizza Bojito-Marrero, 

Kathleen Ruping, Raj Edula 

was very common in both groups (48%). RBV dose reduction 

occurred in 8 Asians (26%) and 20 non-Asians (19%). SAE 

occurred in 5 non-Asian and all were unrelated to treatment. 

Conclusion: Asian and non-Asian CHC-1 patients treated with 

SMV+SOF showed similar high treatment response and good 

tolerability, despite high rates of advanced disease and prior 

treatment failure. Anemia and fatigue were common in both 

Asians and non-Asians treated with SOF+RBV. 

1182 

Similar Tolerability and Effectiveness with SMV+SOF 

Therapy in Asians and Non-Asians with Chronic Hepatitis 

C Genotype 1 (CHC-1) but Anemia and Fatigue were 

Common with SOF+RBV in Both Groups 

Christine Y. Chang 1 , Nghia H. Nguyen 2,1 , Changqing Zhao 1,3 , 

Glen A. Lutchman 1 , Aijaz Ahmed 1 , Tami Daugherty 1 , Gabriel 

Garcia 1 , Radhka Kumari 1 , W. Ray Kim 1 , Huy N. Trinh 4 , Vinh D. 

Vu 1 , Winston Ku 1 , My T. Nguyen 4 , Andrew Huynh 4 , Mindie H. 


University Medical Center, Palo Alto, CA; 2 Department of 

Medicine, University of California, San Diego, San Diego, CA; 

3 Institute of Liver Disease, Shugang Hospital, Department of Cirrhosis, 

Shanghai, China; 4 San Jose Gastroenterology, San Jose, CA 

Purpose: Asian CHC-1 patients can have much higher SVR 

with IFN-based therapy but more likely to develop anemia than 

non-Asians. Our goal is to investigate for possible ethnic-related 

differences in SVR12 or tolerability with DAA-based therapy. 

Methods: We evaluated outcomes of DAA-based therapy 

(SOF+SMV, SOF+RBV, or SOF+LDV±RBV for 8-24) in 401 

patients (330 non-Asians and 71 Asians) seen at 2 US centers 

from 12/2013-12/2014. Results: Asians were mostly Vietnamese 

(38%) and Chinese (24%). Non-Asians were mostly 

Caucasian (62%) and Hispanic (26%). Compared to non- 

Asians, Asians were older (mean age: 63±11 vs. 60±9 years, 

p=0.004) with lower BMI (26±7 vs. 28±5 kg/m 2 , p=0.01) 

and more likely to have HCV-2 (30% vs. 19%) or HCV-6 (8% 

vs. 0%), and less likely to have HCV-1 (54% vs. 68%) or 4 (0% 

vs. 1%), p


sis. 14 patients were treated after liver transplantation, 2 of 

those with a cholestatic recurrence of HCV, one with advanced 

graft failure. 1 patient was treated post-kidney transplantation. 

Results: Efficacy: At end of treatment (EOT), 97% (160/165) 

were HCV negative, while 89% (92/103) reached SVR 12. 

Subgroup analysis: HCV negativity at EOT: SOF/RBV +/- PEG- 

IFN: 100% (54/54); SOF, DAC +/- RBV 98% (64/65); SOF, 

SMP +/- RBV 90% (27/30); SOF/LED +/- RBV 94% (14/15). 

Subgroup analysis for SVR 12: SOF/RBV +/- PEG-IFN: 93% 

(40/44); SOF, DAC +/- RBV 87% (27/31); SOF, SMP +/- 

RBV 89% (24/27); SOF, LED +/- RBV n.a.. SVR 12 in patients 

with cirrhosis: 74% (23/31), SVR 12 in pretreated patients: 

85% (44/52), SVR 12 in patients after liver transplantation 

75% (6/8). 5 patients discontinued prematurely (3 patients 

died during treatment, 2 patients had a virological relapse 

after premature discontinuation), 6 patients had a virological 

relapse after EOT. No virological breakthrough occurred. 

Safety: 1 patient being treated in the course of an advanced 

graft failure after liver transplantation died due to complications 

of transplant failure and re-transplantation during treatment. 

2 patients with advanced cirrhosis died during treatment 

(sepsis n=1, right heart failure n=1). Conclusions: New DAA 

are highly effective with respect to EOT, and SVR 12. Groups 

of patients with previous treatment failure, after liver transplantation, 

or cirrhosis achieve significantly higher SVR rates than 

with previous treatments. However, in “real-life”, patients with 

advanced liver cirrhosis still seem to be “difficult to treat” with 

respect to treatment efficacy. Since near all patients will have 

reached SVR 12 until September, 2015, uni - and multivariate 

analyses will be presented at the AASLD Liver Meeting 2015. 

Disclosures: 

The following authors have nothing to disclose: Christoph R. Werner, Daniel P. 

Egetemeyr, Nisar P. Malek, Ulrich M. Lauer, Christoph P. Berg 

1184 

Sofosbuvir Combination Therapy and Chronic Hepatitis 

C Infection: Is Cultural Background a Predictor of Sustained 

Virologic Response? 

Amber Tierney, John R. Lake, Amar Mahgoub, Carolyn Schmitt, 

Qi Wang, Mohamed A. Hassan; University of Minnesota, Minneapolis, 

MN 

Background: Treatment for hepatitis C infection has rapidly 

evolved over the past few years. Although sustained virologic 

response (SVR) rates in clinical trials are high, it is unknown 

if this response translates accurately to clinical practice. Our 

aim was to detail our initial experience treating patients with 

hepatitis C infection with sofosbuvir containing regimens with 

an emphasis on genotype 4 infection. Methods: A retrospective 

chart review was performed on all patients treated for 

hepatitis C infection from December 2013 to May 2015 at a 

single academic center with a large population of East African 

immigrants. Results: During the study period, 254 patients were 

treated for hepatitis C infection. Treatment regimens included 

either sofosbuvir/ribavirin (RBV) +/- pegylated interferon (PEG- 

IFN), sofosbuvir/simeprevir or ledipasvir/sofosbuvir +/- RBV. 

179 patients had 3 month follow-up after treatment with an 

overall SVR rate of 79%. Advanced fibrosis was associated with 

relapse in genotype 1 infection (p=0.02). Prior treatment, viral 

load, and duration of treatment did not have a statistically significant 

effect on SVR (p=0.64, 0.40, and 0.23, respectively). 

Successful response to treatment was associated with improvement 

in alanine and aspartate aminotransferases (p


(14/42) were female, 21% (9/42) were Hispanic, and 9% 

(4/42) were Black. The median time from transplant to treatment 

initiation was 5.4 years (IQR: 2.1-8.8 years). Thirteen 

patients experienced one or more episodes of hepatic decompensation 

and/or SAE. Anemia requiring transfusion, the most 

common event, occurred in 8/13 (62%), while 7/13 (54%) 

decompensated. The cumulative incidence of hepatic decompensation/SAE 

was 31% [95% CI, 16-41%]. Risk factors for 

decompensation/SAE included pre-treatment hemoglobin [OR 

= 0.61 per g/dL; 95% CI: 0.40-0.88; p


All DAAs regimen were analyzed together. For TN and TE 

patients, 2 analyses evaluated the RBV use (regardless of the 

treatment duration and for a fixed 12 weeks treatment duration) 

and 3 analyses evaluated the treatment duration (regardless 

of, with, or without RBV). Another analysis compared 12 

wks with RBV vs. 24 wks without RBV. Meta-analyses were 

performed according to the Der Simonian and Laird method 

and reported weight-adjusted SVR12 gains. Results: 10 RCTs 

(1307 GT1 cirrhotic patients (573 TN, 734 TE)) were selected. 

The distribution of DAA combinations was as follows: SIM+SOF 

(COSMOS, N=168), 3D Abbvie (TURQUOISE II, N=380), 

DCV-TRIO BMS (UNITY 2, N=202), GPV+EBV (C-WOR- 

THY, N=253), and LDV+SOF (6 RCTs, N=496). In G1 TN 

cirrhotic patients, the use of ribavirin (ΔSVR=+2.45%;95%CI: 

-1.27 to +6.16%,NS,N=411) or an extended duration 

(ΔSVR=+0.64%;95%CI:-3.03 to +4.32%,NS,N=434) did not 

increase SVR12. In GT1 TE cirrhotic patients, the use of ribavirin 

did not increase SVR12 (ΔSVR=+0.23%;95%CI:-3.54 

to +4.01%,NS,N=494), even in analyses restricted to 

12wks treatment duration (ΔSVR=+1.85%;95%CI:-3.75 to 

+7.44%, NS,N=267). Conversely, extended duration was 

associated with higher SVR rates (ΔSVR=+6.35%;95%CI: 

+1.49 to +11.20%,p=0.018,N=532), even with ribavirin 

use (ΔSVR=+8.26%;95%CI: +2.29 to +14.23%, p=0.028, 

N=278). The magnitude of SVR gain was similar between the 

different DAA regimen suggesting that a 18 wks extended 

duration (as performed in the C-WORTHY study) would be 

sufficient for other combos. Only four studies (229 patients) 

compared 12wks +RBV vs. 24wks w/o RBV. Extended duration 

was associated with a 5.09% SVR gain that did not reach 

significance, probably because of a type II error and the overweight 

of the SIRIUS study. Conclusion: In GT1 TN patients with 

compensated cirrhosis, RBV as well as extending treatment 

duration are useless. In GT1 TE patients with compensated 

cirrhosis, RBV does not increase SVR rates but extending treatment 

duration (18 or 24 wks) significantly increases SVR rates 

Disclosures: 




Thong Dao - Board Membership: Schering-Plough, Schering-Plough, Schering-Plough, 

Schering-Plough; Speaking and Teaching: Roche, Gilead, Roche, 

Gilead, Roche, Gilead, Roche, Gilead 




Odile Goria - Speaking and Teaching: Gilead, Janssen 


MSD, BMS 

The following authors have nothing to disclose: Carine Richou, Jean Paul Cervoni, 

Delphine Weil, Claire Vanlemmens, Anne Minello, Christine Silvain, Thierry 

Thevenot 

1188 

Real World Effectiveness Of Sofosbuvir Based Therapy 

In Chronic HCV Infected Patients: The Results From Data 

Anlaysis Of 167 Patients In Qatar 

Moutaz F. Derbala 1 , Aliaa Amer 2 , Saad R. Alkaabi 1 , Nazeeh 

Z. Dweik 1 , Khaleel H. Sultan 1 , Yasser M. Kamel 1 , Khalid Al-Ejji 1 , 

Hamid Wani 1 , Fuad I. Pasic 1 ; 1 GASTROENTEROLOGY & HEPA- 

TOLOGY, Hamad Hospitaland Weill Cornell Medical College in 

Qatar,Consultant Gastroenterology & Hepatology HMC, Doha, 

Qatar; 2 Hematopathology, Laboratory Medicine and pathology, 

Hematology Section, Hamad Medical Corporation, Doha, Qatar 

Background and Aims: However, limited data were available; 

Sofosbuvir (SOF) was approved in Qatar in early 2014. In both 

interferon (IFN) based triple therapy and IFN-free combinations, 

sofosbuvir have been used to treat different patient populations 

with various liver disease severity and comorbidities. Our aim 

is to assess the effectiveness of SOF based therapy in a real 

world setting. Methods: In the Qatar HCV registry, all patients 

who started on the following treatment regimens were included 

in the analysis: SOF/ribavirin (RBV), SOF/daclatasvir (DCV), 

SOF/simeprevir (SMV), and SOF/PegIFN/RBV. Demographics, 

history of liver disease were collected, and laboratory tests 

were performed at baseline. Clinic, adverse events and virological 

data were collected throughout treatment and post-treatment 

follow-up. Results: 167 patients were treated with SOF 

based therapy. 36%of the patients had cirrhosis and 52.8% 

were previously treated with PR or Protease inhibitor (PI) +PR. 

14.1% of the patients underwent liver transplantation.13.8 % 

patients were treated with SOF+PegIFN/RBV triple therapy. 

The overall SVR4 rate was 96%, 92%, and 87% in patients 

treated within genotype 1, 4 and 3 respectively. The overall 

SVR4 rate differed according to the prior history and fibrosis. 

The SOF based therapy achieved 89% SVR4 rate in cirrhotic 

patients and a 97% SVR4 in non-cirrhotic patients. One patient 

discontinued all medications due to viral breakthrough with no 

discontinuation due to adverse events. The most common side 

effects are Anemia and neutropenia, headache and myalgia 

in both SOF/RBV and SOF/Peg-IFN/RBV groups. Conclusions: 

SOF based Therapy is safe and effective in the treatment of 

HCV Genotypes 1, 2, 3 and 4 in a real world setting. However, 

both SOF/RBV and SOF/Peg-IFN/RBV regimens are less 

reliable in cirrhotic patients and genotype 3 

Demographic Data Of The Studied Patients 

Disclosures: 

The following authors have nothing to disclose: Moutaz F. Derbala, Aliaa Amer, 

Saad R. Alkaabi, Nazeeh Z. Dweik, Khaleel H. Sultan, Yasser M. Kamel, Khalid 

Al-Ejji, Hamid Wani, Fuad I. Pasic 

1189 

IFN and/or RBV-Free Therapy (Rx) with Simeprevir+Sofosbuvir 

(SMV+SOF) was Well-Tolerated and Effective 

for Chronic Hepatitis C Genotype 1 (CHC-1) Including 

Post-Liver Transplant (LT) and Decompensated Non-Liver 

Transplant (Non-LT) Patients: A Single-Center Experience 

Glen A. Lutchman 1 , Nghia H. Nguyen 1,3 , Christine Y. Chang 1 , 

Aijaz Ahmed 1 , Tami Daugherty 1 , Gabriel Garcia 1 , Radhka 

Kumari 1 , W. Ray Kim 1 , Soumi Gupta 4 , Dilesh Doshi 2 , Mindie H. 


University Medical Center, Palo Alto, CA; 2 Health Economics 

& Outcomes Research, Janssen Scientific Affairs, Titusville, NJ; 

3 Department of Medicine, University of California, San Diego, San 

Diego, CA; 4 Medical Affairs, Janssen Scientific Affairs, Titusville, 

NJ 

Purpose: In the U.S., HCV is the leading cause of liver-related 

death and CHC-1 is the most common type. Until recently, 

effective and safe Rx options were limited. Our goal is to 

evaluate outcomes of SMV+SOF Rx in a diverse single-center 

patient cohort from 12/2013/12/2014. Methods: A total of


234 consecutive patients were identified, of which 36 were 

excluded due to prior SOF-based or current IFN-based Rx or 

pending SVR12 (n=25), yielding 198 for study inclusion: 50 LT 

and 148 non-LT. Results: Mean age was ~60 and the majority 

(80%) in both groups were Caucasian or Hispanic. In non-LT, 

10% had liver cancer, 71% cirrhosis (84% had decompensation 

with clinical symptoms or MELD≥10 or CPT≥7), while only 

8% of LT group had cirrhosis. In LT group, SVR12 was similar 

for both GT 1a (88%) and 1b (87%), and in cirhrosis (87%) 

but lower in those with prior-Rx (73%) (Fig 1). In non-LT group, 

SVR12 was higher for GT 1b, noncirrhosis, Rx-naïve compared 

to 1a, cirrhosis, and prior-Rx, though none of these differences 

were statistically significant (Fig 1). Rx was well tolerated in 

all groups with 6 Rx-unrelated SAEs. On multivariate analysis, 

also inclusive of sex and age, the presence of at least 2 difficult-to-treat 

features (prior Rx, cirrhosis, or GT 1a) was a significant 

negative predictor for SVR12 (HR=0.06, CI 0.01-0.73, 

p=0.027), but not the presence of just one of such factors or LT. 

Conclusions: SMV+SOF Rx (no RBV) was safe and effective for 

both LT and non-LT patients with SVR12 of ~80% for those with 

difficult-to-treat features and 90% for those without. Significant 

predictor for lower SVR12 was presence of 2 or more of such 

features. 

Disclosures: 




Inc. 

Gabriel Garcia - Board Membership: Health Metrics Systems 



Dilesh Doshi - Employment: Janssen 





The following authors have nothing to disclose: Glen A. Lutchman, Nghia H. 

Nguyen, Christine Y. Chang, Tami Daugherty, Radhka Kumari, Soumi Gupta 

1190 

Virological and Clinical Response in Patients with 

HCV-Related Mixed Cryoglobulinemia Treated with 

Interferon-Free Regimens: Preliminary Results of a Prospective 

Pilot Study. 

Laura Gragnani, Alessia Piluso, Teresa Urraro, Alessio Fabbrizzi, 

Elisa Fognani, Luisa Petraccia, Monica Monti, Anna Linda Zignego; 

Dept. Experimental and Clinical Medicine, University of 

Florence, Firenze, Italy 

Background: Hepatitis C virus (HCV) infection is often associated 

with extrahepatic manifestations: the most frequent is 

represented by mixed cryoglobulinemia (MC). MC is an autoimmune/B-cell 

lymphoproliferative disorder, characterized by 

the presence of circulating immune complexes, called cryoglobulins. 

In 5–30% of cases, MC patients showed symptoms due 

to a systemic vasculitis of small/medium size vessels (mixed 

cryoglobulinemia syndrome, MCS). The etiologic therapy was 

considered the first-line option in MCS patients. However, interferon 

(IFN)-based therapy is frequently not tolerated. Recently, 

the introduction of direct acting antivirals (DAAs) substantially 

changed the treatment of HCV infection, allowing IFN-free 

regimens. No data at present exist about the use of IFN-free 

regimens in MC patients. Our aim was to analyze the efficacy 

and safety of new generation DAAs in IFN-free regimens in MC 

patients. Methods: Pilot prospective study, including 17 patients 

with HCV-associated MC with or without symptoms treated 

with new generation DAAs in IFN-free regimens. Patients were 

treated, for 12 or 24 weeks, with different all-oral antiviral 

regimen: ombitasvir, paritaprevir+ritonavir and dasabuvir ± 

ribavirin; sofosbuvir plus daclatasvir and sofosbuvir plus ribavirin. 

Results: Patients were divided in two different cohorts as follows: 

(a) 10 patients (3 [27.3%] males, mean age 64.73±6.6 

yrs) in the MCS-HCV group; (b) 7 patients (4 [57.1%] males, 

mean age 55.57±10.52 yrs) in the MC-HCV group. The main 

clinical baseline manifestation of MCS included purpura (80%), 

arthralgia (80%), asthenia (100%), peripheral neuropathy 

(90%), renal involvement (40%) and cutaneous ulcers (30%). 

Regarding previous anti-HCV treatment, 7 (41%) patients were 

naïve, 6 (35%) were partial or non responders, 1 (6%) patient 

was virological relapser and 3 (18%) discontinued previous 

treatment for severe adverse events. At week 8 of treatment, all 

patients were HCV RNA negative. Furthermore, a reduction of 

the cryocrit values was evident in all cases (p


1191 

Evaluation of the efficacy and safety to dual oral therapy 

with daclatasvir and asunaprevir in Japanese reallife 

settings 

Hitomi Sezaki 1 , Fumitaka Suzuki 1 , Tetsuya Hosaka 1 , Shunichiro 

Fujiyama 1 , Hideo Kunimoto 1 , Yushi Sorin 1 , Yusuke Kawamura 1 , 

Norio Akuta 1 , Masahiro Kobayashi 1 , Yoshiyuki Suzuki 1 , Satoshi 

Saitoh 1 , Yasuji Arase 1 , Kenji Ikeda 1 , Mariko Kobayashi 2 , Hiromitsu 

Kumada 1 ; 1 Hepatology, Toranomon Hospital, Tokyo, Japan; 

2 Research institute for hepatology, Toranomon Hospital, Kawasaki, 

Japan 

Aim: The first IFN-free dual oral therapy in Japan with daclatasvir 

(DCV) and asunaprevir (ASV) for patients with chronic 

HCV-1b infection was approved since Sep. 2014. Since the 

real-world population is heterogeneous, it is important to indicate 

treatment outcomes in patients excluded from clinical trials 

(CTR). The aims of this study were to evaluate the efficacy 

and safety to 24-week DCV/ASV therapy in Japanese reallife 

settings. Methods: 604 GT1b patients started dual oral 

therapy with DCV/ASV until Jan. 31, 2015 in our hospital. 

423 patients achieved end of treatment (EOT) until May 16. 

180 of 423 patients met Phase3-CTR inclusion criteria and 

243 patients unmet it. This criteria is as following; age ≤ 75 

yr, absence of HCC, Hb ≥ 8.5g/dL, platelet counts ≥ 50 x 

10 3 /mL, albumin ≥ 3.5 g/dL, HCV RNA levels ≥ 5 logIU/mL, 

and absence of prior DAAs therapy. We evaluated SVR rates 

after treatment and the adverse events (AE) during therapy. 

Results: CTR-unmet patients were older than CTR-met patients 

(median 73 yr vs. 65 yr, p


1193 

ITPA polymorphisms are predictors of anemia but no 

contribution to outcomes in chronic hepatitis C patients 

who treated with interferon plus ribavirin 

Xiumei Chi 1 , Mo-li Wang 3 , Yu Pan 1 , Jing Jiang 2 , Hongqing Yan 1 , 

Ruihong Wu 1 , Xiaomei Wang 1 , Junqi Niu 1 ; 1 Hepatology, The first 

hospital of Jilin University, Changchun, China; 2 Epidemoology, 

The first hospital of Jilin University, Changchun, China; 3 The fourth 

hospital of Jilin University, Changchun, China 

Background and Aims: In a genome-wide association study of 

patients being treated for chronic hepatitis C (HCV), two single 

nucleotide polymorphisms (SNPs) within or adjacent to the 

inosine triphosphatase (ITPA) gene (rs1127354, rs6051702) 

were shown to protect against ribavirin induced hemolytic anemia 

during early stages of treatment. We then evaluated these 

polymorphisms/variants as predictors of anemia or treatment 

outcomes (SVR) in a large cohort of naive HCV patients who 

were treated with interferon plus ribavirin. Methods: Genetic 

material was available from 351 HCV patients. These patients 

were enrolled in the first hospital of Jilin University who underwent 

interferon plus ribavirin combination therapy and they had 

no history of any other interferon-based therapy. rs6051702 

in C20orf194, rs1127354 in ITPA and rs8099917 in IL-28B 

were genotyped and tested for their association with hemoglobin 

(Hb) reduction during treatment and with treatment 

outcomes. A stepwise multivariate regression analysis was 

performed to identify factors associated with outcome of the 

therapy. Results: The ITPA rs1127354 minor variants were 

significantly associated with protection against reduction Hb 

level at week 4 (p=2.65×10 -4 during HCV combination therapy. 

SNP rs6051702 was not associated with the hemoglobin 

decline to >3 g/dL at week 4 in our study (p=0.861). 

rs1127354 was strongly and independently associated with 

protection from quantitative Hb reduction at week 4. The minor 

alleles of each variant protected against HB reduction. The 

ITPase deficiency genotype was associated with lower rates of 

anemia over the entire treatment period (72 weeks). No association 

with sustained virological response (SVR) was observed. 

Conclusion: ITPA polymorphisms influences hemoglobin levels 

during ribavirin combination therapy. rs1127354 was conrmed 

to be a useful predictor of ribavirin-induced anemia in 

HCV patients but not a predictive factor for treatment outcomes. 

Disclosures: 

The following authors have nothing to disclose: Xiumei Chi, Mo-li Wang, Yu Pan, 

Jing Jiang, Hongqing Yan, Ruihong Wu, Xiaomei Wang, Junqi Niu 

1194 

Cardiac arrhythmia in patients with chronic hepatitis C 

virus infection treated with sofosbuvir-including regimen 

Hélène Fontaine 1,2 , Arnaud Lazarus 3,4 , Caroline Pecriaux 5 , 

François Bagate 3 , Philippe Sultanik 1,2 , Estelle Boueyre 1,2 , Marion 

Corouge 1,2 , Vincent Mallet 1,2 , Anaïs Vallet-Pichard 1,2 , 

Philippe Sogni 1,2 , Denis Duboc 3 , Stanislas Pol 1,2 ; 1 Hepatology 

Unit, Cochin Hospital, Paris, France; 2 Université Paris Descartes, 

INSERM USM20, Institut Pasteur et Assistance Pubique - Hôpitaux 

de Paris, Paris, France; 3 Cardiology Unit, Hôpital Cochin, Assistance 

Publique -Hôpitaux de Paris, Paris, France; 4 Rhythmology 

Unit, Clinique Ambroise Paré, Neuilly Sur Seine, France; 5 Unité de 

Pharmacovigilance, Hôpital Cochin, Assistance Publique - Hôpitaux 

de Paris, Paris, France 

Introduction: Treatment of chronic hepatitis C virus (HCV) 

infection has dramatically improved since the advent of direct 

anti-viral agents, including Sofosbuvir. Sustained virological 

response, which corresponds to a viral cure, is achievable 

in more than 90 % of patients. To date, several thousands of 

patients have been treated with Sofosbuvir-based regimens 

worldwide. Since their approval in France, oral anti-viral therapies 

were prescribed for patients with complications, including 

cirrhosis and life-threatening extra-hepatic manifestations. 

Methods: We analyzed prospectively the incidence of arrhythmias 

and conduction disorder in a cohort of patients treated 

by Sofosbuvir (new antiviral HCV) in our hospital from January 

2, 2014 to December, 31, 2014. Results: We observed five 

cases (1.2 %) of severe arrhythmia, including three cardiac 

conduction defects, within the first days of Sosfobuvir-based 

therapy among 415 patients who had been treated in our 

unit. There were two females and three males. The median 

age was 58 (50-75) years. Two patients were co-infected with 

HIV. Arrhythmia relapsed when Sofosbuvir was reintroduced in 

one patient. Among the 3 conduction disorders, two patients 

were treated with beta-blocker and another with amiodarone. 

A pacemaker was implanted in all 3 patients with conduction 

disorders. The three patients who underwent a pacemaker 

implantation were not pacemaker-dependent on the controls 

after stopping antiviral treatments. Conclusion: Sofosbuvir 

seems to have cardiac toxicity. The mechanisme underlying 

Sofosbuvir cardiotoxicity has to be determined. A particular 

attention should be given to patients with a medical history of 

rhythm or conduction abnormalities, including careful monitoring 

(e.g., Holter ECG recordings) of cardiac rhythm during the 

first days of Sofosbuvir therapy. 

Disclosures: 

Hélène Fontaine - Board Membership: Gilead, BMS, Janssen, Abbvie; Speaking 

and Teaching: Gilead, BMS, Janssen, Abbvie, MSD 

Arnaud Lazarus - Board Membership: Boston Scientific, Medtronic; Consulting: 

Sorin Group; Employment: Biotronik 


Roche 

Philippe Sogni - Board Membership: BMS, Gilead; Consulting: Gilead, Roche, 

MSD, BMS, Janssen, Mayoli-Spindler 

Stanislas Pol - Board Membership: Sanofi, Bristol-Myers-Squibb, Boehringer Ingelheim, 

Tibotec Janssen Cilag, Gilead, Glaxo Smith Kline, Roche, MSD, Novartis; 

Grant/Research Support: Glaxo Smith Kline, Gilead, Roche, MSD; Speaking and 

Teaching: Sanofi, Bristol-Myers-Squibb, Boehringer Ingelheim, Tibotec Janssen 

Cilag, Gilead, Glaxo Smith Kline, Roche, MSD, Novartis 

The following authors have nothing to disclose: Caroline Pecriaux, François 

Bagate, Philippe Sultanik, Estelle Boueyre, Marion Corouge, Vincent Mallet, 

Denis Duboc 

1195 

Utility of acoustic radiation force impulse (ARFI) for predicting 

treatment response of patients with chronic hepatitis 

C receiving triple therapy with protease inhibitor 

plus pegylated interferon and ribavirin 

Ryoko Yamada 1 , Naoki Hiramatsu 1 , Tsugiko Oze 1 , Ayako Urabe 1 , 

Yuki Tahata 1 , Naoki Morishita 1 , Takayuki Yakushijin 1 , Yuichi 

Yoshida 1 , Tomohide Tatsumi 1 , Yasuharu Imai 2 , Tetsuo Takehara 1 ; 

1 Department of Gastroenterology and Hepatology, Osaka University 

Graduate School of Medicine, Suita, Japan; 2 Ikeda Municipal 

Hospital, Ikeda, Japan 

Background & Aim: With interferon-based therapy, liver fibrosis 

has been reported to correlate with the virologic response 

in patients with chronic hepatitis C (CH-C). Acoustic radiation 

force impulse (ARFI) is known as a non-invasive procedure for 

assessment of liver fibrosis. In this study, we investigated the 

utility of ARFI for CH-C patients receiving triple therapy with 

protease inhibitor (PI) plus pegylated interferon (Peg-IFN) and 

ribavirin (RBV). Patients & Methods: A total of 105 genotype 1 

CH-C patients were enrolled. They started triple therapy (telaprevir/simeprevir, 

26/79) between February 2011 and June 

2014, following liver biopsy and ARFI assessment. The base-


line characteristics were as follows: age, 60.9 ± 10.0 years 

old; median HCV RNA, 6.6 log IU/mL; platelet count, 15.8 ± 

5.1 x10 4 /mm 3 ; ALT, 56.4 ± 42.0 IU/L; Vs value, 1.47 ± 0.46 

m/sec; Fibrosis score (F1/2/3/4), 55/18/15/10. Single 

nucleotide polymorphisms located near the IL28B (rs8099917) 

gene were determined using a real-time PCR system. Results: 

Vs values at baseline showed positive correlation with Fibrosis 

score (p


wave PI, telaprevir (TVR)-based triple therapy of patients with 

genotype 1b HCV infection. Methods: In this study, 23 patients 

infected with HCV genotype 1b who had not achieved sustained 

virologic response (SVR) after TVR-based therapy (17 

relapsers, 1 non-responder and 5 patients who discontinued 

treatment) received SMV-based therapy at Osaka University 

Hospital and institutions participating in the Osaka Liver Forum. 

Among them, 19 patients had experienced Peg-IFN plus RBV 

therapy (9 relapsers and 10 non-responders) before TVR-based 

therapy. Resistance testing of the NS3 region was performed 

by ultra-deep sequencing at the start of SMV-based therapy. 

Results: In per-protocol, the SVR12 rates were 52% (11/21), 2 

patients were non-responders and 8 patients relapsed. The only 

factor that was significantly associated with SVR was the virologic 

response to Peg-IFN plus RBV (relapser vs. non-responder, 

OR: 0.048, p = 0.024, SVR rates: 88% vs 25%) in univariate 

analysis, while that to TVR-based therapy had no association 

with SVR. Ultra-deep sequencing at the start of SMV-based 

therapy revealed that substitution at the R155 position, which 

is cross-resistant to TVR and SMV, was detected in only one 

patient with low frequency (R155G, 0.4%) and this patient 

achieved SVR. RAVs of D168 at the start of SMV-based therapy 

were detected in four patients with low frequency (~ 1%). The 

SVR rates of patients with and without RAVs of D168 were 50% 

(2/4) and 53% (9/17). In two non-response patients for SMVbased 

therapy, emergent TVR-resistant variants, V36G (92.2%) 

and T54A (82.9%), were undetectable at the start of SMVbased 

therapy, but de novo variants of D168 were detected 

during the therapy. Conclusions: In this study, no correlation 

was observed between baseline RAVs and the effect of SMVbased 

therapy. The efficacy of retreatment with SMV-based 

therapy after failure of TVR-based therapy was suggested to 

depend on previous virologic response to Peg-IFN plus RBV. 

Disclosures: 






The following authors have nothing to disclose: Naoki Morishita, Naoki Hiramatsu, 

Tsugiko Oze, Yuki Tahata, Ryoko Yamada, Takayuki Yakushijin, Akira 

Yamada, Eiji Mita, Toshihiko Nagase, Yukinori Yamada, Toshifumi Ito, Masahiko 

Tsujii, Masami Inada, Yasuharu Imai, Ryotaro Sakamori, Yuichi Yoshida, 

Tomohide Tatsumi, Norio Hayashi 

1198 

Better Work Productivity and Activity in Patients on 

Ombitasvir/ Paritaprevir/Ritonavir and Dasabuvir with 

or without Ribavirin (3D+RBV or 3D) in Treatment-Naïve 

Adults with Genotype 1(GT1) Chronic Hepatitis C 

Yan Liu 1 , Yan Luo 1 , Xuan Liu 1 , Danielle Sullivan 1 , Timothy R. 

Juday 1 , Brian Conway 2 ; 1 Abbvie Inc, North Chicago, IL; 2 Vancouver 

Infectious Diseases Centre, Vancouver, BC, Canada 

Background Interferon-containing regimens for treatment of 

Chronic Hepatitis C can further worsen the work productivity 

and activity impairment (WPAI) of patients. We report the 

WPAI of the 311 patients who participated in the phase 3b 

MALACHITE-I study. Methods GT1a-infected patients were randomized(2:1) 

to receive 12 weeks of 3D+RBV or 12 weeks 

of Telaprevir plus peginterferon/ribavirin(TPV+pegIFN+RBV) 

and 12-36 additional weeks of pegIFN/RBV. GT1b-infected 

patients were randomized(2:2:1) to 3D+RBV, 3D, or TPV+pegIFN/RBV. 

Baseline and change from baseline during treatment 

and post treatment(PT) periods were measured using 

the WPAI instrument (hepatitis C module, WPAI-HCV) for all 

five treatment groups The statistical significance of differences 

between treatment groups in mean change from baseline to 

Week 12 and to PT Week 12 was assessed by subgenotype, 

respectively. Results The analyses included 69 patients on 

3D+RBV and 34 on TPV+pegIFN/RBV in GT1a, and 84 on 

3D+RBV, 83 on 3D and 41 on TPV+pegIFN/RBV in GT1b. 

Mean baseline overall work impairment(OWI) and activity 

impairment scores for Week 12 and PT Week 12 analyses 

varied from 2.5 to 9.1 and 9.3 to 13.4 across treatment 

groups. Mean changes from baseline in WPAI-HCV scores 

are summarized in the Table. Numerically smaller additional 

impairments were observed in OWI at Week 12 with 3D+RBV 

in GT1b(p=0.004) and GT1a(p


pharmacokinetics studies,H2-blocker famotidine (40 mg q12 

hours) & PPI omeprazole (20 mg single dose) decreased LDV 

Cmax by 17-20% & 4-11% respectively but had no effect on 

AUC 1 . In the ION trials, which evaluated the effect of SOF/ 

LDV in genotype 1 patients, patients were not allowed to be 

on any acid-suppressing agents for 28 days before initiation of 

therapy. Hence, currently there is no clinical data on the effect 

of acid suppressing agents on SVR in patients treated with 

SOF/LDV. Aim: To evaluate the effect of co-administration of 

PPI (at doses recommended in the package insert) with LDV/ 

SOF on SVR12 in hepatitis C genotype 1 patients. Methods: 

Data from a combined treatment cohort of three Hepatology 

referral centers were obtained on all hepatitis C genotype1 

patients on PPI treated with SOF/LDV with or without ribavirin 

(RBV). Treatment duration (8-24 weeks) & addition of RBV was 

at the discretion of the treating hepatologist Results: Of the 50 

patients in this cohort,30 (60%) were male, 35 (70%)had genotype 

1a, 29 (58%) were Caucasians, & 68% had cirrhosis. 

Mean age was 58 years, with a mean BMI of 31 kg/m 2 . 25 

(50%) were treatment naïve & the remaining were treatment 

experienced with peg-interferon (peg-IFN) + RBV with or without 

Telaprevir/Bocepravir, peg-IFN + RBV + SOF, Simeprevir 

+SOF with or without RBV, SOF+RBV. PPI was co-administered 

at the recommended package insert dosing primarily 

for GERD (86%). 22/43 had undetectable viral load after 4 

weeks of treatment initiation, & 92% (22/24) who completed 

treatment achieved end-of treatment (EOT) response defined by 

undetectable viral load. Treatment was very well tolerated with 

treatment discontinuation in only one patient due to intractable 

nausea & vomiting requiring hospitalization. SVR12 results to 

follow Conclusion: The co-administration of a PPI with LDV/ 

SOF at recommended doses does not seem to effect EOT 

response. Symptoms of GERD are also generally well tolerated 

at the package insert PPI dosing while on treatment 1 German 

P, et al. Effect of food & acid reducing agents on the relative 

bioavailability & pharmacokinetics of ledipasvir/sofosbuvir 

fixed dose combination tablet.15th International Workshop on 

Clinical Pharmacology of HIV & Hepatitis Therapy; Washington, 

D.C. 2014 

Disclosures: 

Hector E. Nazario - Advisory Committees or Review Panels: Gilead, Janssen; 

Speaking and Teaching: Gilead, Merck, Abbvie, Janssen 

Manjushree Gautam - Speaking and Teaching: Gilead 

Stevan A. Gonzalez - Consulting: AbbVie; Speaking and Teaching: Gilead, 

Salix, AbbVie 

Ruben Ramirez-Vega - Speaking and Teaching: Gilead 

Apurva A. Modi - Speaking and Teaching: Salix, Merck, Gilead, Abbvie, Janssen 

The following authors have nothing to disclose: Jamil S. Alsahhar, Mohammad 

Ashfaq, Milka Ndungu, Olga Teachenor, Gabriel R. Gonzales 

1200 

Pre-Existing Co-Morbidities and Co-Medications of 

Patients Undergoing Treatment of Chronic HCV G1 

Infection in German Real-Life 

Peter Buggisch 1 , Hanns F. Loehr 2 , Gerlinde Teuber 3 , Hermann 

Steffens 4 , Michael R. R. Kraus 5 , Peter R. Geyer 6 , Bernd Weber 7 , 

Thomas Witthoeft 8 , Uwe Naumann 9 , Elmar Zehnter 10 , Dagmar 

Hartmann 11 , Bernd Dreher 11 , Manfred Bilzer 11 ; 1 IFI Liver Center 

Hamburg, Hamburg, Germany; 2 Gastroenterological Practice, 

Wiesbaden, Germany; 3 Gastroenterological Practice, Frankfurt, 

Germany; 4 Practice of Internal Medicine, Berlin, Germany; 

5 Department II, Klinikum Burghausen, Burghausen, Germany; 

6 Gastroenterological Practice, Fulda, Germany; 7 Competence 

Center Addiction, Kassel, Germany; 8 Gastroenterological Practice, 

Stade, Germany; 9 Center of Medicine, Berlin, Germany; 10 Gastroenterological 

Practice, Dortmund, Germany; 11 MSD Pharma 

GmbH, Haar, Germany 

Background: Information about co-morbidities of patients (pts) 

currently treated for chronic HCV genotype 1 (G1) infection in 

real-life is scarce. The present interim analysis of the NOVUS 

observational study was therefore aimed to investigate the 

frequency of pre-existing and ongoing co-morbidities of pts 

treated for chronic HCV G1 infection in German real-life and 

to determine the frequency of co-medications. Methods: From 

April 2012 until January 2014, 536 pts with HCV G1 infection 

were recruited in the ongoing NOVUS study by 97 practices 

and hospitals in Germany. Until now, pre-existing co-morbidities 

before triple therapy of HCV G1 infection with boceprevir 

(BOC) were documented for 469 pts. Results: Ongoing 

co-morbidities were reported for 329 of 469 pts (70%) before 

treatment of HCV G1 infection. Overall, 599 ongoing co-morbidities 

(multiple answers allowed) were documented. The most 

frequently reported co-morbidities were obesity (BMI >30 kg/ 

m 2 ) (19%), cardiovascular diseases (18%), psychiatric disorders 

(14%), opiate substitution (13%), gastrointestinal diseases 

(11%), metabolic disorders (8%), thyroid gland diseases (7%), 

bone and joint diseases (6%), skin diseases (5%), HIV- co-infection 

(4%) and kidney diseases (2%). When co-morbidities 

were analyzed by gender (female vs. male), thyroid diseases 

occurred more frequently in females (13% vs. 3%, P


Michael R. R. Kraus - Advisory Committees or Review Panels: Merck/MSD, 

Roche, Gilead, BMS, Janssen, ABBVIE; Consulting: Merck/MSD, Roche; Speaking 

and Teaching: Merck/MSD, Gilead, BMS, Janssen, ABBVIE 

Bernd Weber - Consulting: Mundipharma, Indivior, Hexal, Janssen, Bristol Myers 

Squibb, AbbVie, MSD, Gilead Sciences 

Uwe Naumann - Speaking and Teaching: MSD, Roche, BMS, Abbott, VIIV, Janssen, 

Boehringer Ingelheim, Gilead 

Dagmar Hartmann - Employment: MSD Germany 

Bernd Dreher - Employment: MSD 

Manfred Bilzer - Consulting: MSD Germany 

The following authors have nothing to disclose: Hanns F. Loehr, Hermann Steffens, 

Peter R. Geyer, Thomas Witthoeft, Elmar Zehnter 

1201 

High ribavirin dose and extended treatment can raise 

sustained virologic response in chronic hepatitis C genotype 

1 patients treated with simeprevir, pegylated interferon 

plus ribavirin 

Yuki Tahata 1 , Naoki Hiramatsu 1 , Tsugiko Oze 1 , Ayako Urabe 1 , 

Naoki Morishita 1 , Ryoko Yamada 1 , Takayuki Yakushijin 1 , Yukiko 

Saji 2 , Yoshinori Doi 3 , Hideki Hagiwara 4 , Akira Kaneko 5 , Hiroyuki 

Fukui 6 , Masami Inada 7 , Kazuhiro Katayama 8 , Atsuo Inoue 9 , Yuichi 

Yoshida 1 , Tomohide Tatsumi 1 , Norio Hayashi 4 , Tetsuo Takehara 1 ; 

1 Department of Gastroenterology, Osaka University Graduate 

School of Medicine, Suita, Japan; 2 Itami City Hospital, Itami, 

Japan; 3 Otemae Hospital, Osaka, Japan; 4 Kansai Rosai Hospital, 

Amagasaki, Japan; 5 NTT West Osaka Hospital, Osaka, Japan; 

6 Yao Municipal Hospital, Yao, Japan; 7 Toyonaka Municipal Hospital, 

Toyonaka, Japan; 8 Osaka Medical Center for Cancer and 

Cardiovascular Diseases, Osaka, Japan; 9 Osaka General Medical 

Center, Osaka, Japan 

Background and Aim: Combination therapy with simeprevir 

(SMV), pegylated interferon (Peg-IFN) plus ribavirin (RBV) is 

recommended as first-line interferon-based therapy for patients 

with chronic hepatitis C (CH-C) genotype 1 in Japan. However, 

the factors associated with a sustained virologic response (SVR) 

have not been fully examined. In this study, we investigated 

factors associated with SVR in patients with CH-C treated with 

SMV, Peg-IFN plus RBV combination therapy. Patients and 

Methods: This multicenter study was conducted at Osaka University 

Hospital and institutions participating in the Osaka Liver 

Forum. A total of 531 patients with CH-C treated with SMV, 

Peg-IFN plus RBV were enrolled in this study (244 males, 287 

females; mean age: 62.2 ± 10.0 y.o.; 229 naïve patients, 212 

Peg-IFN plus RBV-experienced patients). SVR was evaluated by 

HCV-RNA negativity at 12 weeks after the end of treatment 

(SVR12). As a rule, naïve patients and prior relapsers were 

treated for 24 weeks, and patients with non-response (NR) to 

previous therapies were treated for 24 or 48 weeks (24 weeks: 

64 patients, 48 weeks: 23 patients). Results: The discontinuance 

rate due to adverse effect was 8.0% (42/523). In per-protocol, 

the SVR12 rates were 87% for naïve patients, 95% for 

relapsers and 42% for non-responders. Among naïve patients, 

IL28B genotype (rs8099917, TT vs. non-TT, OR: 0.044, p = 

0.001) and RBV dose (< 10/10-12/≥ 12 mg/kg/day, OR: 

4.513, p = 0.041) were significantly associated with SVR on 

multivariate analysis including baseline factors and drug adherence. 

In patients with IL28B TT genotype, the SVR12 rates were 

high regardless of RBV dose (93-100%), while in patients with 

IL28B non-TT genotype, stratified analysis of RBV dose showed 

that SVR12 rates were affected by the RBV dose (p = 0.046), 

being 44% (8/18) for patients given < 10 mg/kg/day of RBV, 

78% (14/18) for those given 10-12 mg/kg/day of RBV and 

100% (3/3) for those given ≥ 12 mg/kg/day of RBV. In NR 

patients with HCV-RNA negativity at 12 and 24 weeks, the 

SVR12 rates were significantly higher in patients treated for 

48 weeks than those treated for 24 weeks (88%, 15/17 vs. 

58%, 25/43; p = 0.03). As for SVR12 rates according to the 

timing of HCV-RNA disappearance, the SVR12 rates of 24- 

and 48-week treatment were 91% (10/11) and 100% (1/1) in 

patients with undetectable HCV-RNA at 2 weeks, 38% (9/24) 

and 89% (8/9) at 4 weeks, 43% (6/14) and 83% (5/6) at 

5-12 weeks, respectively. Conclusion: Higher RBV dose for 

naïve patients with unfavorable IL28B genotype and extended 

treatment for NR patients can raise SVR in CH-C genotype 

1 patients treated with SMV, Peg-IFN plus RBV combination 

therapy. 

Disclosures: 

Kazuhiro Katayama - Consulting: Nobel pharma; Speaking and Teaching: 

Bayer, MSD, Otsuka pharmaceutical Co.,Ltd., Glaxo, Zeria pharmaceutical Co., 

Ltd., Bristol Meyers 





The following authors have nothing to disclose: Yuki Tahata, Naoki Hiramatsu, 

Tsugiko Oze, Ayako Urabe, Naoki Morishita, Ryoko Yamada, Takayuki Yakushijin, 

Yukiko Saji, Yoshinori Doi, Hideki Hagiwara, Akira Kaneko, Hiroyuki Fukui, 

Masami Inada, Atsuo Inoue, Yuichi Yoshida, Tomohide Tatsumi, Norio Hayashi 

1202 

Hepatitis C Cure Results in a Modest, but Durable, 

Decrease in Liver Stiffness 

Sweta Chekuri, Kian Bichoupan, Sanders Chang, Neal M. Patel, 

Ponni V. Perumalswami, David P. Del Bello, Rachna Yalamanchili, 

Alyson Harty, Douglas Dieterich, Andrea D. Branch; Division of 

Hepatology, Mount Sinai, New York, NY 

Background & Aims: We investigated the impact of HCV cure 

on liver stiffness (LS). Methods: FibroScan was used to measure 

LS of 61 patients treated with an interferon (IFN)-containing or 

IFN-free regimen (2008-2015) who achieved a sustained virological 

response (SVR24). Scores ≥ 14 kPa denoted cirrhosis. 

Mann-Whitney and Wilcoxon signed-rank tests were used for 

analysis. Results: The median age was 60 years (± 11). The 

median pre-treatment LS score was 9.1 kPa (IQR: 6.1 - 15 kPa), 

the median post-SVR score was 7.3 kPa (IQR: 5.3 – 10 kPa), 

and the median change was -2.1 kPa (IQR: -5.3 – -0.4 kPa), a 

modest, but statistically significant decrease, p


C trough 

levels were higher in patients with more advanced liver 

fibrosis; they correlated particularly strongly with type 4 collagen 

7S, WFA + -M2BP, and FIB-4 levels. ASV C trough 

prediction 

of hepatic transaminase elevations or VBT was difficult, and 

elevated ASV C trough 

levels were not observed even in patients 

with compromised renal function. 

Disclosures: 

Itaru Ozeki - Speaking and Teaching: BMS 

Disclosures: 

Kian Bichoupan - Consulting: Janssen Pharmaceuticals, Gilead Sciences 


Jannsen, Acaria Pharmacy, Abbvie 




The following authors have nothing to disclose: Sweta Chekuri, Sanders Chang, 

Neal M. Patel, Ponni V. Perumalswami, David P. Del Bello, Rachna Yalamanchili 

1203 

Clinical Significance of Blood Asunaprevir Concentrations 

Levels during Combined Daclatasvir and Asunaprevir 

Therapy 

Itaru Ozeki; Department of Gastroenterology, Sapporo Kosei General 

Hospital, Sapporo, Japan 

[Objectives] This study evaluated the clinical significance of 

blood asunaprevir (ASV) concentrations levels during combined 

daclatasvir+ASV therapy in patients with genotype 1 

chronic hepatitis C. [Subjects and Methods] The study included 

116 patients (36 males [31.0%]; median age, 73 years) starting 

this combination therapy. Liver cirrhosis (LC) was present 

in 62 (53.4%) patients. The median estimated glomerular filtration 

rate (eGFR) was 65.5 mL/min/1.73 m 2 . ASV C trough 

levels 

were analyzed by background variables, and then analyzed 

according to the presence/absence of hepatic transaminase 

elevations, virological breakthrough (VBT), and renal dysfunction. 

Liver fibrosis was determined using Fibroscan ® EI values as 

well as FIB-4 and Wisteria floribunda agglutinin-positive human 

Mac-2-binding protein (WFA + -M2BP) levels. Serum WFA + - 

M2BP levels were measured with HISCL ® M2BPGi reagent (Sysmex 

Co., Kobe, Japan). ASV C trough 

levels were measured using 

HPLC in patients receiving blood sampling 8-12 hours after 

oral administration of ASV 1 week after starting ASV treatment. 

Hepatic transaminase elevations was defined as cases higher 

than twice the upper limit of the normal range. Each parameter 

was expressed as the median. [Results] 1) Analysis of ASV 

C trough 

levels by background variables revealed that the levels 

did not vary with age, gender, body mass index, or eGFR, but 

it was significantly higher in the LC group (119 ng/mL; n=62) 

than in the non-LC group (52 ng/mL; n=54) (p=0.001) . 2) 

Analysis of the correlation of ASV C trough 

levels with factors possibly 

associated with liver fibrosis (platelet, albumin, hyaluronic 

acid, type 4 collagen 7S, FIB-4, WFA + -M2BP, and FS EI values) 

revealed that all were significant (p












The following authors have nothing to disclose: Bernhard Scheiner, Philipp 

Schwabl, Sebastian Steiner, David Chromy, Theresa Bucsics, Berit A. Payer, 

Maximilian C. Aichelburg, Armin Rieger, Katharina Grabmeier-Pfistershammer 

1205 

Real-world effectiveness of ledipasvir/sofosbuvir 8 

weeks chronic hepatitis C treatment 

Peter Buggisch 1 , Karsten Wursthorn 1 , Albrecht Stoehr 1 , Aline 

Gauthier 2 , Petar K. Atanasov 2 , Joerg Petersen 1 ; 1 Asklepios Klinik 

St. Georg Haus L, IFI Institut für Interdisziplinäre Medizin, Hamburg, 

Germany; 2 Amaris Consulting, London, United Kingdom 

Background and Aims: Ledipasvir/Sofosbuvir (LDV/SOF) single 

tablet regimen (STR) is approved in Europe for chronic 

hepatitis C (CHC) patients with genotypes (GT) 1, 3 and 4. In 

the ION-3 study 8 weeks (8w) of LDV/SOF was non-inferior 

to 12w in previously untreated GT1 patients without cirrhosis 

with no benefit for the addition of ribavirin (RBV). According 

to the summary of product characteristics (SmPC) 8w may 

be considered in treatment naïve, non-cirrhotic patients with 

HCV RNA< 6million IU/mL. The aim of the present analysis 

is to characterise the population receiving 8w LDV/SOF and 

to describe outcomes in clinical practice. Methods: The first 

CHC patients treated with 8w LDV/SOF in a single centre in 

Germany, and for whom SVR12 will be available for presentation, 

were included. Baseline characteristics, prior treatment 

history, safety and effectiveness were investigated. Analysis 

was performed using descriptive statistics. Results: 86 patients 

met the inclusion criteria and initiated 8w treatment with LDV/ 

SOF between 21/11/2014 to 07/04/2015. No patient 

had ribavirin added to the STR. The mean (SD) age was 49.8 

(12.1) years and 44.2% were males. Genotype distribution 

was 52.3%, 45.4% and 2.3% for GT1a, GT1b and GT4. The 

METAVIR stage distribution at baseline was 51.2%, 26.7%, 

17.4% and 4.7% for F0, F1, F2 and F3. No patients had cirrhosis. 

Median HCV RNA at baseline was 794,328 (Q1-Q3: 

169,824-2,041,738, Min-Max: 11 -18,620,871) IU/ml, two 

patients had HCV RNA ≥ 6 million IU/mL (11,481,536 and 

18,620,871 IU/ml, both METAVIR stage F0). 4.7% of patients 

were HIV co-infected and no patients were HBV co-infected. 

Overall, 99% of the patients were treatment-naïve. One patient 

had relapsed after previous interferon (INF)/RBV therapy. To 

date, one patient discontinued therapy due to lack of adherence. 

Two patients experienced adverse events: one assessed 

as unrelated and one as possibly related to treatment. Among 

patients for whom data is available, SVR4 was 100% (69/69) 

and SVR12 was 100% (14/14). Complete results for SVR12, 

adverse events and discontinuations will be available at the 

time of presentation. Conclusion: 8w LDV/SOF is predominantly 

prescribed according to the SmPC for treatment-naïve 

non-cirrhotic CHC patients with HCV RNA


Disclosures: 



The following authors have nothing to disclose: Eduardo A. Rodriguez, Cynthia 

Levy, Kalyan R. Bhamidimarri 

1207 

Assessment of serum hepatitis C virus RNA 24 weeks 

after the end of treatment using TaqMan PCR is more 

relevant than after 12 weeks for predicting sustained 

virological response by simeprevir- versus telaprevir-based 

triple therapy 

Tatsuo Kanda, Shingo Nakamoto, Reina Sasaki, Yuki Haga, 

Masato Nakamura, Shin Yasui, Makoto Arai, Fumio Imazeki, 

Osamu Yokosuka; Department of Gastroenterology and Nephrology, 

Chiba University, Graduate School of Medicine, Chiba, 

Japan 

Background and Aims: Hepatitis C virus (HCV) infection causes 

acute and chronic hepatitis and results in cirrhosis and hepatocellular 

carcinoma (HCC). Direct-targeting antiviral agents 

(DAAs) against HCV with or without peginterferon plus ribavirin 

result in higher rates of eradication of this virus and shorter 

treatment duration. We examined which is better for predicting 

persistent virological response, the assessment of serum HCV 

RNA 12 or 24 weeks after the end of treatment for predicting 

sustained virological response (SVR12 or SVR 24, respectively) 

in patients treated with peginterferon plus ribavirin and HCV 

NS3/4A protease inhibitor. Patients and Methods: In all, 145 

Japanese patients infected with HCV genotype 1 treated with 

peginterferon plus ribavirin with HCV NS3/4A protease inhibitor 

were retrospectively analyzed: 59 patients (57.6 years; 

male, 42; and IL28B rs8099917TT, 40) and 86 patients (60.8 

years; male, 42; and IL28B rs8099917TT, 58) were treated 

by telaprevir- and simeprevir-including regimens, respectively. 

Written informed consent was obtained from all patients, and 

this study conformed to the ethical guidelines of the 1975 

Declaration of Helsinki and was approved by the Ethics Committee 

of Chiba University, School of Medicine (No.523 and 

No.1462). HCV RNA was measured by TaqMan HCV Test, 

version 2.0, real-time PCR assay. IL28B rs8099917 was analyzed 

by TaqMan SNP assay. SVR12 and SVR24, respectively, 

were defined as HCV RNA negativity at 12 week and 24 week 

after stopping treatment. Results: Total SVR rates were 78.0% 

and 70.7%, in telaprevir and simeprevir groups, respectively. 

In telaprevir group, SVR rates of treatment-naïve, previous-treatment 

relapsers and others were 76.7%, 87.0% and 50.0%, 

respectively. In simeprevir group, SVR rates of treatment-naïve, 

previous-treatment relapsers and others were 75.0%, 100% 

and 25.0%, respectively. In telaprevir group, all 46 patients 

with SVR12 finally achieved SVR24. In simeprevir group, only 

4 (7.1%) of total 56 patients with SVR12 achieved SVR24, and 

these 4 patients were all previous-treatment relapsers (67.5 

years; male, 2; and IL28rs8099917TT, 2). Conclusion: SVR12 

were suitable for predicting persistent virological response in 

almost cases. In simeprevir-including regimens, SVR12 could 

not always predict persistent virological response. Clinician 

should use SVR24 for predicting persistent virological response 

in the use of DAA treatment for a group of real-world patients 

chronically infected with HCV. 

Disclosures: 

Tatsuo Kanda - Grant/Research Support: MSD K.K.; Speaking and Teaching: 

AJINOMOTO PHARMACEUTICALS CO., LTD, CHUGAI PHARMACEUTICAL 

CO., LTD, GlaxoSmithKlein, BMS, Jansen, Daiichisankyo; Stock Shareholder: 

Mitsubishi Tanabe Pharma 

Osamu Yokosuka - Grant/Research Support: Chugai, Taiho, Bristol Myers, 

Takeda 

The following authors have nothing to disclose: Shingo Nakamoto, Reina Sasaki, 

Yuki Haga, Masato Nakamura, Shin Yasui, Makoto Arai, Fumio Imazeki 

1208 

Reduced Survival in Elderly Liver Transplant Recipients: 

How Old is Too Old? 

Nae-Yun Heo 1,2 , Ajitha Mannalithara 1 , Prowpanga Udompap 1 , 

Donghee Kim 1 , Waldo Concepcion 3 , Carlos O. Esquivel 3 , W. 

Ray Kim 1 ; 1 Division of Gastroenterology and Hepatology, Stanford 

University School of Medicine, Stanford, CA; 2 Department 

of Internal Medicine, Inje University College of Medicine, Busan, 

Korea (the Republic of); 3 Department of Surgery, Stanford University 

School of Medicine, Stanford, CA 

Background/Aims: Liver transplantation (LTx) is increasingly 

performed in patients over 65 years of age, an age group 

traditionally considered to be high risk. Prognostic models specifically 

designed to predict post-LTx outcome in elderly patients 

may help select optimal candidates. Methods: Records of all 

adult (>18 years) LTx recipients were extracted from the Organ 

Procurement and Transplantation Network (OPTN) database 

for 2004-2013. After exclusion of recipients of multi-organ or 

repeated LTx, recipients were divided into three groups:


1209 

The Small Molecule TLR9 Antagonist COV08-0064 

Protects from Warm Ischemia Reperfusion Injury of the 

Liver 

Mohamed E. Shaker 1,2 , Bobby N. Trawick 3 , Wajahat Z. Mehal 1 ; 

1 Section of Digestive Diseases, Yale School of Medicine, New 

Haven, CT; 2 Department of Pharmacology & Toxicology, Faculty of 

Pharmacy, Mansoura University, Mansoura 35516, Egypt; 3 Center 

for Organic Chemistry, Mallinckrodt Pharmaceuticals, St. Louis, 

MO 

Warm ischemia/reperfusion (IR) injury is a major reason for 

failure of liver surgeries, and also limits the use of steatotic livers 

for transplantation. A significant portion of the tissue injury 

in IR is mediated by activation of an innate immune response 

upon reperfusion. Minimizing this innate immune response is an 

attractive goal to limit IR injury. Here, we present a novel and 

selective small molecule Toll-like receptor (TLR9) antagonist with 

the ability to protect from IR injury. Methods: Male C57Bl/6 

mice (8-10 weeks) were subjected to 1h of warm hepatic ischemia 

by occlusion of the vasculature supplying the left and 

median lobes of the liver. Control mice had the same surgical 

procedure, but without interruption of liver blood flow. COV08- 

0064 was administered at a dose of 80 mg/kg intraperitoneally 

1h prior starting ischemia. At 3 h and 12 h after resuming 

blood flow, the livers were examined by standard histology, 

scored for injury, and qPCR for pro- and anti-inflammatory cytokines 

performed. The ability of COV08-0064 to regulate inflammatory 

cytokine production was also tested in vitro on mouse 

macrophages and plasmacytoid dendritic cells (pDCs). Results: 

COV08-0064 pretreatment for I/R-mice resulted in limiting the 

rise in serum alanine aminotransferase activity (595 ± 124 vs 

3233 ± 785, P


Cox proportional hazards analysis to assess post-transplant 

survival. Results Between 2002 and 2014, the mean age of 

liver transplant registrants increased from 51.2 to 55.7, with 

a more prominent increase in HCV-positive (50.9 to 57.9) 

than HCV-negative (51.3 to 54.3) registrants. The proportion 

of registrants aged ≥ 60 years increased from 19% to 41% 

and the proportion aged ≥ 65 years increased from 8.1% to 

17%. Almost identical trends were observed among liver transplant 

recipients. Among transplant registrants, increasing age 

was associated with increased mortality before transplantation 

(adjusted sub-hazard ratio 1.30 for age 50-59, 1.49 for age 

60-64, 1.73 for age 65-69 and 2.04 for age ≥70 relative 

to age 18-49) and decreased likelihood of transplantation. 

Among transplant recipients, increasing age was associated 

with increased post-transplant mortality (adjusted hazard ratio 

1.16 for age 50-59, 1.34 for age 60-64, 1.61 for age 65-69 

and 1.87 for age ≥70 relative to age 18-49). Conclusions 

Dramatic aging of liver transplant recipients and registrants 

occurred from 2002 to 2014, most prominently in HCV-infected 

patients. The combination of higher waitlist mortality and 

higher post-transplant mortality in elderly patients suggests that 

listing of these patients should be pursued cautiously. 

Proportion of liver transplant registrants and recipients >=60 

years of age by HCV status, 2002-2014 

Grade 1 [47(38.2%)], ACLF Grade 2 [47(38.2%)], and ACLF 

Grade 3 [29(23.6%)]. Results: ACLF patients were younger 

(52yrs. Vs 54 yrs., p=0.018) and have significantly higher 

MELD score at LT (30.47±6.98 vs. 19.74±4.44, p=0.001). 

Higher proportion of underlying liver disease included alcohol, 

autoimmune hepatitis and Wilson’s disease in ACLF patients. 

ACLF recipients needed longer hospitalization (20.13±23.87 

vs. 11.51±11.41 days, p=0.001), ICU stays (9.43±17.91 

vs. 4.23±8.99 days, p=0.001), higher early (≤90 days) readmissions 

(0.51±0.73 vs. 0.35±0.68, p=0.008), and higher 

early (≤90 days) surgical re-explorations (26 vs 14.8%, 

P=0.002). Graft loss was significant in the ACLF group (35.6 

% vs. 27.5%, p=0.044, median follow up =1476 days), and 

was pronounced at 1 year follow up (18.7 % vs. 10.3%, 

p=0.009). Graft (p=0.035, Figure 1A) and patient survival 

(p=0.025, Fig.1B) was significantly lower in the ACLF group, 

with incremental poor graft (p=0.013, Fig.1A) and patient 

survival (p=0.022, Figure 1B) in higher grades of ACLF. Graft 

(p=0.086, Fig. 1C) and patient survival (p=0.118, Fig. 1D) 

based on MELD severity categories were not significant by 

Kaplan-Meier analysis. Conclusion: ACLF recipients defined by 

EASL-CLIF score have inferior overall graft and patient survival, 

and their length of post-LT hospitalization, length of ICU stay, 

early readmissions, and early surgical re-explorations were 

significantly higher. 

Figure 1. Cumulative graft (1A) and patient survival (1B) in ACLF 

grades. Cumulative graft (1C) and patient survival (1D) in various 

MELD categories. 

Disclosures: 


The following authors have nothing to disclose: Feng Su, Lei Yu, Kristin Berry, Iris 

W. Liou, Jorge Reyes, Stephen C. Rayhill, George N. Ioannou 

1212 

Outcomes of acute-on-chronic liver failure in liver transplant 

recipients based on EASL-CLIF consortium definition 

Anuj Sharma 1 , William J. Goldkamp 1 , Satheesh Nair 2,1 , Jason 

Vanatta 2,1 , James Eason 2,1 , Sanjaya K. Satapathy 2,1 ; 1 University 

of Tennessee Health Science Center, Memphis, TN; 2 Methodist 

Transplant Institute, Memphis, TN 

Background: Outcomes of liver transplant (LT) recipients with 

acute-on-chronic liver failure (ACLF) with recently defined 

EASL-CLIF consortium definition has not been studied. We 

aimed to study the long term outcomes of LT patients with 

ACLF. Methods: After retrospective review of 833 consecutive 

LT recipients (04/2006 to 03/2013), and exclusion of 

patients without chronic liver disease, re-LTs, SLK recipients, 

and acute liver failure, 123 ACLF and 582 without ACLF were 

included. LT recipients with ACLF were divided into ACLF 

Disclosures: 

Satheesh Nair - Advisory Committees or Review Panels: Jansen; Grant/Research 

Support: Gilead; Speaking and Teaching: Abbvie, Valeant, BMS 

Sanjaya K. Satapathy - Advisory Committees or Review Panels: Gilead, Intercept; 

Grant/Research Support: Genfit, Gilead, Biotest 

The following authors have nothing to disclose: Anuj Sharma, William J. Goldkamp, 

Jason Vanatta, James Eason


1213 

Prediction of Posthepatectomy Liver Failure based on 

Preoperative Liver Fibrosis Assessment: Comparison of 

Mac-2 Binding Protein Glycosylation Isomer and Liver 

Stiffness Measurement 

Takahiro Nishio, Kojiro Taura, Kazutaka Tanabe, Gen Yamamoto, 

Yukihiro Okuda, Yoshinobu Ikeno, Satoru Seo, Kentaro Yasuchika, 

Etsuro Hatano, Hideaki Okajima, Toshimi Kaido, Shinji Uemoto; 

Department of Surgery, Graduate School of Medicine, Kyoto University, 

Kyoto, Japan 

Posthepatectomy liver failure (PHLF) is one of the major 

causes of hepatectomy-related mortality, and accurate prediction 

of PHLF based on preoperative liver function test is 

essential. Recently, Mac-2 binding protein glycosylation isomer 

(M2BPGi) has been proposed as a useful liver fibrosis 

marker, while liver stiffness (LS) measurement was also well 

accepted as a noninvasive assessment tool for liver fibrosis. 

We evaluated the usefulness of these novel liver fibrosis markers, 

M2BPGi and LS, for predicting PHLF. One hundred and 

thirty-eight patients with hepatocellular carcinoma undergoing 

hepatectomy between August 2011 and October 2014 were 

analyzed. Preoperative serum M2BPGi level was measured 

in addition to a routine blood test, and LS was measured by 

acoustic radiation force impulse-based Virtual Touch Tissue 

Quantification (VTTQ) technology. Remnant liver volume rate 

(REM) was calculated by computed tomography volumetry. 

PHLF was diagnosed according to the definition of the International 

Study Group of Liver Surgery and was graded as A, 

B, or C. The accuracy of predictors was assessed by the area 

under the receiver operating characteristic curve (AUROC). The 

mean serum M2BPGi level (cut off index) was 1.0±0.5 in F0-1, 

1.7±1.1 in F2-3, and 4.5±3.6 in F4; and the mean VTTQ value 

(m/s) was 1.4±0.3, 1.7±0.6, and 2.7±0.9, respectively; both 

of which significantly increased according to the progression 

of liver fibrosis stage (p < .01). M2BPGi had AUROCs of 0.80 

for the diagnosis of F≥2, 0.82 for F≥3, and 0.83 for F4, which 

were better than any other fibrosis markers including the VTTQ 

value. PHLF occurred in 34 patients (24.6%): grade A, 15 

patients (10.9%); grade B, 14 patients (10.1%); grade C, 5 

patients (3.6%). The AUROCs of M2BPGi were 0.61 for the 

prediction of PHLF grade ≥A, 0.70 for grade ≥B, and 0.64 

for grade C; while those of VTTQ value were 0.66, 0.77, and 

0.72, respectively. Multivariate stepwise selection identified 3 

best fit significant factors for the prediction of PHLF: M2BPGi 

(odds ratio [OR], 1.6; 95% confidence interval [CI], 1.1-2.4, 

p = .013), platelet count (OR, 0.42; 95% CI, 0.23-0.72, p < 

.01), and REM (OR, 0.37; 95% CI, 0.23-0.59, p < .01). The 

predictive model including these 3 variables had AUROCs 

of 0.77 for predicting PHLF grade ≥A, 0.81 for grade ≥B, 

and 0.80 for grade C; and this model enabled the estimation 

of safe resection liver volume according to the risk of PHLF 

by the preoperative M2BPGi and platelet count. Conclusions. 

M2BPGi and LS were useful markers for predicting PHLF as 

well as liver fibrosis progression. 

Disclosures: 


The following authors have nothing to disclose: Takahiro Nishio, Kojiro Taura, 

Kazutaka Tanabe, Gen Yamamoto, Yukihiro Okuda, Yoshinobu Ikeno, Satoru 

Seo, Kentaro Yasuchika, Hideaki Okajima, Toshimi Kaido, Shinji Uemoto 

1214 

Two-Year Results of a Pilot Program in Early Liver Transplantation 

for Severe Alcoholic Hepatitis 

Brian P. Lee 1 , Po-Hung Chen 1 , Ruben Hernaez 1 , Ahmet Gurakar 1 , 

Benjamin Philosophe 2 , Zhiping Li 1 ; 1 Medicine, Johns Hopkins University 

School of Medicine, Baltimore, MD; 2 Surgery, Johns Hopkins 

University School of Medicine, Baltimore, MD 

OBJECTIVE: Six months of alcohol abstinence is typically 

required before a patient is considered for liver transplant (LT). 

In October 2012, our LT center initiated a pilot program to 

transplant selected patients with acute alcoholic hepatitis. The 

purpose of this study was to assess the results of this pilot program 

and the effects on organ allocation. METHODS: Data 

was collected from all patients with alcohol-related liver disease 

(ALD) since initiation of our pilot program in October 

2012. These patients were stratified into three groups based 

on indication for LT: severe alcoholic hepatitis as first liver 

decompensation (Group 1), alcoholic cirrhosis with ≤6 months 

abstinence (Group 2), alcoholic cirrhosis with ≥6 months 

abstinence (Group 3). Follow-up period was defined as time 

between date of transplant and date of last follow-up or death. 

Recidivism was any evidence of alcohol consumption following 

LT. The number of organs allocated for ALD within 2 years of 

the pilot program was compared to the 2-year period immediately 

prior to implementation. RESULTS: 36 patients underwent 

LT. All patients in Group 1 had Maddrey’s Discriminant 

Function greater than 32. Table 1 has patient characteristics 

and outcomes. 18 of 36 (50%) LTs performed for ALD had 

less than 6 months of abstinence prior to listing. Compared 

to the 2-year period prior to the pilot program, there was an 

increase in the proportion of organs indicated for ALD (20% 

vs. 12%; p=0.13). 6-month survival was 100%, 100%, 83% 

in Group 1, 2, 3, respectively (p=0.16). CONCLUSIONS: For 

carefully selected patients presenting with alcoholic hepatitis, 

early LT resulted in similar 6-month survival and alcohol relapse 

rates compared to patients undergoing LT after 6 months of 

abstinence. Patients transplanted for acute alcoholic hepatitis 

(Group 1) had significantly higher MELD score and shorter 

length of time on the transplant list compared to both other 

groups. The implementation of a policy allowing early LT for 

ALD resulted in an increase in proportion of organs allocated 

for alcohol-related indications, but did not reach statistical significance. 

Table 1: Patient Characteristics and Outcomes 

*Excluding two patients who died during the post-operative 

period 

Disclosures: 

Ahmet Gurakar - Advisory Committees or Review Panels: Gilead; Grant/ 

Research Support: BMS 

The following authors have nothing to disclose: Brian P. Lee, Po-Hung Chen, 

Ruben Hernaez, Benjamin Philosophe, Zhiping Li


1215 

Early Liver Transplantation in Severe Alcoholic Hepatitis: 

A Multi-Center, Retrospective, Cohort Study 

Brian P. Lee 1 , David W. Victor 2 , Darryn R. Potosky 3 , Ibrahim 

A. Hanouneh 4 , Mary E. Rinella 5 , Michael D. Voigt 6 , Sheila L. 

Eswaran 7 , Howard P. Monsour 2 , R. Mark Ghobrial 2 , Jessica A. 

Keppel 7 , Zhiping Li 1 ; 1 Johns Hopkins University School of Medicine, 

Baltimore, MD; 2 Houston Methodist Hospital, Houston, 

TX; 3 University of Maryland School of Medicine, Baltimore, MD; 

4 Cleveland Clinic, Cleveland, OH; 5 Northwestern University Feinberg 

School of Medicine, Chicago, IL; 6 University of Iowa Carver 

College of Medicine, Iowa City, IA; 7 Rush Medical College, Chicago, 

IL 

OBJECTIVE: Liver transplant (LT) centers typically require 6 

months of alcohol abstinence before a patient is considered 

for LT. Severe alcoholic hepatitis (SAH) refractory to medical 

treatment has a 6-month survival estimated at 30%. A European 

study showed that early LT of SAH in a highly selected 

group of patients could improve survival. The purpose of this 

study was to examine the outcomes of early LT for SAH in U.S. 

transplant centers. METHODS: All 111 U.S. LT centers were surveyed 

for patients who underwent LT for acute SAH (Maddrey’s 

Discriminant Function >32) as the first presentation of liver 

decompensation, and listing for LT prior to 6 months of alcohol 

abstinence. 48 centers responded to the survey. 10 centers 

reported such experience, of which 7 submitted retrospective 

data. All patients presented from October 2006 to November 

2014. Alcohol relapse was defined as any alcohol consumption 

following LT. Binge drinking was defined as 6 units of 

alcohol in a day for men, 4 units for women. Frequent drinking 

was defined as any alcohol consumption more than 4 days a 

week. RESULTS: 41 patients underwent early LT for acute SAH. 

Median follow-up was 2.4 years. Median period of alcohol 

abstinence prior to LT listing was 53 days. Median MELD at 

listing was 34. Median Maddrey’s discriminant function was 

69. Only 12 (29%) received steroids. Of those excluded from 

steroid therapy (n=29), 11 (38%) had confirmed infection, 

6 (21%) had presumed infection without confirmed source, 

5 (17%) had GI bleed, 7 (24%) were excluded due to provider 

choice. Six-month and 1-year survival were both 100%. 

Overall survival was 89%. 9 (22%) had alcohol relapse; of 

these patients (n=9), 6 (67%) reported binge drinking, 7 (78%) 

reported frequent drinking, 4 (44%) reported both binge and 

frequent drinking, and 4 (44%) had re-achieved sobriety by 

time of last follow-up. Compared to the cohort of Mathurin et 

al, steroid use was lower (29% vs. 92%, p


Table 1 

Disclosures: 

James W. Ferguson - Advisory Committees or Review Panels: Astellas, Novartis 

The following authors have nothing to disclose: Suman Verma, Fiona M. Hand, 

Matthew J. Armstrong, Marie de Vos, Douglas Thorburn, Terry Pan, John R. 

Klinck, Rachel Westbrook, Georg Auzinger, Andrew Bathgate, Steven Masson, 

Andrew Holt, Diarmaid D. Houlihan 

1217 

Sublingual administration of tacrolimus is a feasible 

route and requires a lower dose to achieve similar drug 

exposition in adult liver transplant recipients. 

Sandra Solari 2 , Alejandra Cancino 1 , Blanca M. Norero 1 , Rodrigo 

Wolff 1 , Francisco Barrera 1 , Alejandro Soza 1 , Marco Arrese 1 , Juan 

Francisco Guerra 3 , Nicolás Jarufe 3 , Jorge A. Martínez 3 , Carlos E. 

Benítez 1 ; 1 Gastroenterology, Pontificia Universidad Católica de 

Chile, Santiago, Chile; 2 Laboratorio Clínico, Pontificia Universidad 

Católica de Chile, Santiago, Chile; 3 Digestive Surgery, Pontificia 

Universidad Católica de Chile, Santiago, Chile 

Introduction: Oral administration of immunosuppressive drugs 

is not always feasible after liver transplantation and parenteral 

administration of tacrolimus (TAC) implies a significantly higher 

expense of resources. The aim of this study is to evaluate the 

feasibility of sublingual (sl) administration of tacrolimus in adult 

liver transplant recipients. Methods: In patients on per oral 

administration (po) serum TAC levels were determined at different 

time points (0, 0.5, 1, 2, 4, 6 and 12 hours), then patients 

were switched to sl administration and TAC dose was tapered 

(starting with a 20-30% reduction of the po dose) to obtain a 

similar trough level that in po administration. Then TAC levels 

were determined at the same time points. The exposition to 

TAC (AUC) was estimated and compared for SL and po administration. 

Results: Seventeen recipients were enrolled. The mean 

time required to taper de sl dose to obtain a proper trough level 

was 16.41±10.5 days (range 6-40 days). There were no differences 

on trough levels on po and sl administrations (7.06±2.2 

vs 7.19±2.0 ng/ml respectively, p=ns). Remarkably, when 

AUC was evaluated, no differences were found in both groups 

(111.4±32.23 ng/ml/h vs 111.4±35.65 ng/ml/h, p=ns). 

However, the dose required to achieve a similar trough level 

was significantly higher when TAC was administered on po vs 

sl route (4.68±2.32 vs 2.94±1.7 mg/d, p=0.018), meaning a 

38% reduction. The maximum concentration after TAC administration 

(Cmax) was similar in sl and po groups (19.65±9.97 

vs 15.88±6.91 ng/ml, p=ns). No severe adverse effects were 

registered on sl group and was well tolerated. In one patient 

very few and small blisters were transiently observed for a few 

days and in 4 patients a spicy taste was transiently observed 

with the meals on sl group. Conclusion: Sublingual administration 

of tacrolimus is a feasible alternative to po administration 

and can also be monitored employing trough levels requiring 

a lower dose to achieve the same drug exposition. These findings 

also suggest that parenteral TAC administration could be 

replaced by sl route. These findings could imply a significantly 

reduction on the cost of immunosuppression therapy. 

Disclosures: 

Alejandro Soza - Advisory Committees or Review Panels: Merck, Roche, Gilead; 

Speaking and Teaching: Merck, BMS, Roche; Stock Shareholder: Gilead, 

Achillion 

The following authors have nothing to disclose: Sandra Solari, Alejandra Cancino, 

Blanca M. Norero, Rodrigo Wolff, Francisco Barrera, Marco Arrese, Juan 

Francisco Guerra, Nicolás Jarufe, Jorge A. Martínez, Carlos E. Benítez 

1218 

Pre-transplant echocardiographic parameters as a tool 

to evaluate post-transplant survival and cardiac outcomes 

in liver transplant recipients 

Daniel W. Bushyhead 1 , James N. Kirkpatrick 2 , David S. Goldberg 

3 ; 1 Internal Medicine, Hospital of the University of Pennsylvania, 

Philadelphia, PA; 2 Cardiology, Hospital of the University of 

Pennsylvania, Philadelphia, PA; 3 Gastroenterology, Hospital of the 


Background Despite advances in liver transplantation and 

pre-operative risk stratification, there remains significant 

post-transplant morbidity and mortality from cardiovascular 

disease. There are limited and conflicting data on the role of 

pre-transplant echocardiography in risk stratifying patients with 

cirrhosis. Moreover, the relationship between pre-transplant 

cardiac function and post-transplant cardiovascular and renal 

disease is unknown. The purpose of our study was to determine 

if pre-transplant echocardiography predicts post-transplant outcomes 

in liver transplant recipients. Methods We conducted a 

retrospective analysis of adult liver transplant recipients at the 

University of Pennsylvania from January 1, 2005 to September 

30, 2014. We included only patients with pre-transplant echocardiograms 

that were read by a cardiologist at our institution. 

Patients with acute liver failure and those without a diagnosis 

of cirrhosis were excluded. Pre-transplant echocardiographic 

parameters were: ejection fraction, diastolic dysfunction, aortic 

stenosis, regional wall motion abnormalities, mitral and tricuspid 

regurgitation, left atrial size, right ventricular size and 

function, pulmonary artery systolic pressure (PASP) and right 

atrial pressure. Post-transplant outcomes were: patient survival, 

major adverse cardiac events (MACE; myocardial infarction 

and heart failure) and chronic kidney disease (CKD) stage 4 or 

5 (estimated glomerular filtration rate


1219 

Reporting Physical Function in UNOS: The Weakness of 

the Karnofsky Performance Status Scale 

Connie W. Wang, John P. Roberts, Jennifer C. Lai; University of 


Background: Currently, physical function (fxn) is reported in 

UNOS using the Karnofsky Performance Status (KPS) scale, 

which ranges from 0-100% as determined by the liver transplant 

(LT) team. Given its subjective nature, we aimed to evaluate 

factors influencing the KPS score in the UNOS registry. 

Methods: Included were all adult LT recipients from 9/08- 

9/13. Physical fxn was categorized as low (0-20%), medium 

(30-70%), or high (80-100%) on the KPS. Cox models assessed 

the relationship between KPS and post-LT mortality, ordinal 

logistic models the relationship between KPS with patient (pt)/ 

center (ctr)-level factors. Results: Of 25,632 LT pts, 21%, 56% 

and 23% had low, medium, and high physical fxn at LT. Low, 

medium, and high physical fxn-ing pts differed by: %female 

(41 vs. 33 vs. 26%), %hospitalized in ICU (54 vs. 4 vs. 1%), 

median lab MELD (35 vs. 20 vs. 15), and median Child score 

(12 vs. 10 vs. 8) [p


1221 

Renal Recovery Outcomes after Liver Transplantation 

in Patients Requiring Pre-operative Renal Replacement 

Therapy 

Rex Cheng 1 , Kimberly Muczynski 2 , Lena Sibulesky 3 , Renuka Bhattacharya 

1 ; 1 Gastroenterology/Hepatology, University of Washington, 

Seattle, WA; 2 Nephrology, University of Washington, Seattle, 

WA; 3 Transplant Surgery, University of Washington, Seattle, WA 

Background: The purpose of this study is to characterize the 

long-term outcomes of renal recovery in liver transplant (OLT) 

recipients who required preoperative renal replacement therapy 

(RRT). Methods: A single center retrospective review was 

performed for OLT recipients who were transplanted from January 

2009 to April 2015 and required RRT at least one day 

prior to transplant. Patients were excluded if they received a 

combined liver-kidney transplant or required a repeat transplant. 

Patient demographics, clinical data, pre- and post-transplant 

laboratory data, number of days on RRT, time to RRT 

cessation were extracted from an electronic medical record. 

Outcomes measured include early renal recovery (cessation 

of RRT within 30 days after OLT), delayed renal recovery (cessation 

of RRT more than 30 days after OLT) or development 

of end-stage renal disease, as well as residual renal function, 

characterized by serum creatinine at 30 and 90 days after 

cessation of RRT. Results: Excluding combined liver-kidney 

transplants (n=17) and patients requiring repeat transplant (n 

= 14), 454 patients underwent liver transplantation during this 

time period. Sixty patients (mean age 51.9 ± 9.7 years, 55.0% 

male, 62% with acute tubular necrosis, mean Model of End- 

Stage Liver Disease score 36.7 ± 4.7 at transplant) received 

RRT prior to OLT. 34 (57%) achieved early renal recovery, 15 

(25%) achieved delayed renal recovery over a mean of 57.6 

days and 11 (18%) were deemed to have end-stage renal 

disease with RRT dependence. Patients who achieved early 

renal recovery had a shorter duration of pre-transplant RRT 

than those who did not, 17 days, [95% confidence interval 

(CI), 12-21 days] versus 26 days [(95% CI, 19-33 days), p = 

0.044]. At 30 days after cessation of RRT (n= 48), 70.5% of 

patients with early renal recovery vs 40.0% of patients with 

delayed renal recovery attained a serum creatinine < 2 (p = 

0.046). At 90 days after cessation of RRT (n=44), 93.3% of 

patients with early renal recovery vs 78.5% of patients with 

delayed renal recovery attained a serum creatinine < 2 (p = 

0.148). Conclusion: Over three-quarters of patients requiring 

pre-transplant RRT achieve renal recovery. Shorter duration 

of RRT is associated with earlier renal recovery. Early renal 

recovery is associated with more rapid improvement in residual 

renal function. The majority of patients who achieve renal 

recovery have improved residual renal function at 90 days 

after RRT cessation. 

Disclosures: 

The following authors have nothing to disclose: Rex Cheng, Kimberly Muczynski, 

Lena Sibulesky, Renuka Bhattacharya 

1222 

Coffee consumption promotes survival and protects 

against hepatocellular carcinoma recurrence following 

liver transplantation: putative mechanisms involve modulation 

of adenosinergic signalling 

Georg Wiltberger 1 , Ruoyu Miao 2 , Undine G. Lange 1 , Rudi 

Ascherl 1 , Hans-Michael Hau 1 , Felix Krenzien 1 , Georgi Atanasov 1 , 

Christian Benzing 1 , Elliot B. Tapper 2 , Linda Feldbrügge 2 , Johannes 

Broschewitz 1 , Michael Bartels 1 , Sven Jonas 3 , Johann Pratschke 4 , 

Yan Wu 2 , Simon C. Robson 2 , Moritz Schmelzle 4 ; 1 Department of 

Visceral, Transplant, Thoracic and Vascular Surgery, University 

Hospital Leipzig, Leipzig, Germany, Leipzig, Germany; 2 Department 

of Medicine, Beth Israel Deaconess Medical Center, Harvard 

Medical School, Liver Center and The Transplant Institute, Div. of 

Gastroenterology, Boston, MA; 3 Department of Hepato-, Pancreato- 

and Biliary Surgery, 310Klinik, Nürnberg, Nürnberg, Germany; 

4 Department of General, Visceral, and Transplant Surgery, 

Charité - Universitätsmedizin Berlin, Campus Virchow Klinikum, 

Berlin, Germany 

Background: Increasing evidence suggests that coffee consumption 

might decrease liver cirrhosis death risk and reduce 

the risk for hepatocellular carcinoma (HCC). Caffeine is a natural 

antagonist to adenosine-mediated receptor signaling and 

has tumoricidal activity in experimental settings. Aim: To determine 

whether coffee consumption decreases HCC recurrence 

and promotes survival following orthotopic liver transplantation 

(OLT) and to examine putative mechanisms of action. 

Methods: Patients who underwent OLT for HCC from January 

2002 to December 2012 were interviewed on the numbers 

of consumed cups of coffee per day before and after OLT. 

Coffee consumption was correlated with patients’ data with 

regard to HCC recurrence and patient survival. For in vitro 

mechanistic assays, human HepG2 HCC cells were analyzed 

for expression of adenosine receptors and ectoenzymes. Cells 

were also treated with adenosine, in the presence or absence 

of 8-(3-Chlorostyryl)-caffeine (CSC) or alloxazine, followed by 

analyses of proliferation, metastasis and alterations in key adenosine-mediated 

cancer pathways inclusive of MAPK (ERK and 

JNK) and NF-kappa B. Results: 90 patients with a median 

age of 60 years (range 73 – 37) and a median BMI of 26.95 

(range 18.3 – 39.4) underwent OLT for histologically confirmed 

HCC. 16 (17.8 %) patients experienced HCC-recurrence after 

median time of 11.5 months (range 1 - 40.5) post-transplant. 

In patients with a high preoperative or postoperative coffee 

intake HCC recurrence was observed less frequently than 

in those with low coffee intake (p=0.018). In a multivariate 

Cox’ proportional hazard model for recurrence-free survival 

only postoperative coffee intake ≥3 cups daily showed a significant 

relevance. HepG2 cells express abundant levels of 

A2A and A2B as well as several key ectoenzymes (including 

ENTPD3, ENTPD5, ENTPD8, CD73, and adenosine deaminase 

1 (ADA1)). These cells show intrinsic capacity to generate 

extracellular adenosine. Exogenous adenosine promotes HCC 

cell growth and metastasis in vitro, which is blocked by CSC, 

but not by alloxazine. Moreover, adenosine induces activation 

of MAPK (ERK and JNK) and NF-kappaB pathways, which can 

be prevented by antagonism of A2A and A2B receptors. Conclusions: 

Coffee consumption is associated with a decreased 

risk of HCC recurrence and increased survival following OLT. 

Experimental data suggest that these results might be, at least 

in part, associated with the antagonist activity of caffeine on 

adenosine-A2AR mediated growth-promoting effects in HCC 

cells. If validated further, our findings have implications for 

future recommendations for HCC patients after OLT.


Disclosures: 





Moritz Schmelzle - Grant/Research Support: Novartis GmbH 

The following authors have nothing to disclose: Georg Wiltberger, Ruoyu Miao, 

Undine G. Lange, Rudi Ascherl, Hans-Michael Hau, Felix Krenzien, Georgi Atanasov, 

Christian Benzing, Elliot B. Tapper, Linda Feldbrügge, Johannes Broschewitz, 

Michael Bartels, Sven Jonas, Johann Pratschke, Yan Wu 

1223 

Recipient obesity is an independent risk factor for 

unplanned reoperations and prolonged operative time 

in orthotopic liver transplant 

Eric S. Wise 1 , Malena Outhay 2 , Michael Maggart 2 , Saad Rehman 

2 , Sir Norman T. Melancon 2 , James Leathers 2 , Kristina Jacomino 

2 , Michelle Izmaylov 2 , Amol Utrankar 2 , Kyle M. Hocking 1 , Sunil 

K. Geevarghese 1,3 ; 1 Surgery, Vanderbilt, Baltimore, MD; 2 School 

of Medicine, Vanderbilt, Nashville, TN; 3 Transplant Center, Vanderbilt, 

Nashville, TN 

Background: Selection of orthotopic liver transplant recipients 

is increasingly inclusive of patients with higher body-mass indices 

(BMI), as BMI has not been implicated as a determinant 

of patient or graft survival. As larger patients present a surgical 

challenge, we aim to characterize the influence of obesity 

on important, yet ill-characterized surgery-specific outcomes. 

Methods: 382 included OLT patients with BMI values reflecting 

obesity (>30 kg/m 2 ) or normal weight (20-25 kg/m 2 ) from 

2005-2014, from a single instituiton, were reviewed for rationally 

chosen recipient-based preoperative factors potentially 

influencing the need for an unplanned reoperation (UR), or 

the length of operation. Factors included BMI, demographics, 

serum laboratory studies, comorbidities, etiology and subjective 

measures of disease severity (e.g. ascites, inpatient status). 

Factors that trended (P30 kg/m 2 (P=.07), younger age 

(P=.04), higher MELD (P=.05) and recent renal replacement 

(P=.08). Younger age (OR 0.97/year, P=.01) and BMI>30 

kg/m 2 (OR 1.72, P=.03) remained independently significant. 

Conclusions: Obesity may make OLT more technically challenging, 

and thus represents an independent risk factor for UR 

and prolonged operative time. 

Disclosures: 

The following authors have nothing to disclose: Eric S. Wise, Malena Outhay, 

Michael Maggart, Saad Rehman, Sir Norman T. Melancon, James Leathers, 

Kristina Jacomino, Michelle Izmaylov, Amol Utrankar, Kyle M. Hocking, Sunil 

K. Geevarghese 

1224 

Ursodeoxycholic Acid as Adjunctive Therapy to Endoscopic 

Management of Non Anastomotic Biliary Lesions 

after Liver Transplant: Effectiveness Depends on Etiology 

Mark Pedersen 1 , David W. Backstedt 2 , Bobby Kakati 2 , Myunghan 

Choi 3 , Anil B. Seetharam 4 ; 1 Internal Medicine, University of 

Arizona College of Medicine-Phoenix, Phoenix, AZ; 2 Gastroenterology, 

University of Arizona College of Medicine-Phoenix, Phoenix, 

AZ; 3 Arizona State University College of Nursing and Health 

Innovation, Phoenix, AZ; 4 Banner Transplant and Advanced Liver 

Disease Center, University of Arizona College of Medicine, Phoenix, 

AZ 

Rationale: Non anastomotic biliary lesions (NABLs) are a cause 

of morbidity after liver transplant (LT) and are postulated to 

occur as a result of diverse mechanisms including insufficiency 

of allograft arterial supply and occult cytomegalovirus (CMV) 

infection. Ursodeoxycholic acid (UDCA) abrogates bile duct 

injury in chronic cholestatic conditions and study is needed 

to evaluate utility as adjunctive therapy for NABLs after LT. 

Aim: Evaluate effect of post-operative UDCA use on markers of 

cholestasis and management of NABLs after LT. Methods: Retrospective, 

unmatched cohort study identifying consecutive LT 

recipients over a 5 year period (2009-2014) with 1 year follow 

up. Recipients with Roux-en-Y biliary reconstruction excluded. 

Biliary complications were defined as endoscopically assessed 

non anastomotic stricture or leak. “UDCA group” was defined 

as subjects with uninterrupted usage of UDCA (300mg PO 

TID) for a minimum of one month post-LT. “Non-UDCA group” 

had no recorded exposure after LT. Baseline demographics 

included: age, gender, indication for LT, Model for End Stage 

Liver Disease (MELD) score at LT, blood type, body mass index 

(BMI), cytomegalovirus (CMV) donor status, need for take-back 

surgery for non biliary complication 1 month from LT, warm 

ischemic time, cold ischemic time, and donation after cardiac 

death (DCD). Outcomes compared between UDCA and non- 

UDCA groups included: number of biliary complications, number 

of ERCPs, number of stents placed, and serum bilirubin at: 

hospital discharge post-LT, first ERCP, 3, 6, 9, and 12 months. 

Categorical variables were analyzed using Fisher’s exact tests. 

Continuous variables were compared with student’s t test with 

p


The following authors have nothing to disclose: Mark Pedersen, David W. Backstedt, 

Bobby Kakati, Myunghan Choi 

The following authors have nothing to disclose: Craig Haifer, Lijia Yu, Julie Pavlovic, 

Paul Gow, Kumar Visvanathan, Adam Testro 

1225 

Quantiferon-CMV testing after cessation of antiviral 

therapy predicts late CMV reactivation after Liver Transplantation 

Siddharth Sood 2,1 , Craig Haifer 1 , Lijia Yu 3 , Julie Pavlovic 1 , Paul 

Gow 1 , Robert M. Jones 1 , Kumar Visvanathan 3 , Peter W. Angus 1 , 

Adam Testro 1 ; 1 Department of Gastroenterology and Liver Transplant 

Unit, Austin Health, Heidelberg, VIC, Australia; 2 Department 

of Gastroenterology & Hepatology, Royal Melbourne Hospital, 

Melbourne, VIC, Australia; 3 Innate Immunity Laboratory, University 

of Melbourne, Melbourne, VIC, Australia 

Introduction CMV is the most common infection to reactivate 

following liver transplantation (OLT). Although antivirals are 

effective in preventing reactivation, some patients develop late 

CMV after prophylaxis or treatment has ceased. Quantiferon– 

CMV (QFN-CMV, Qiagen, USA) measures immune response 

towards CMV. We have previously shown that early non-reactive 

(n.r.) QFN-CMV (suggesting insufficient immunity) at 2 

weeks post-OLT predicts early CMV reactivation, usually within 

the first 2 months post-OLT. This study investigates whether 

n.r. QFN-CMV at 6 months (M6) is associated with late CMV 

after antiviral cessation. Methods 75 OLT recipients were 

monitored as part of a prospective blinded study. Absolute 

QFN-CMV1000 copies/mL). Treatment 

or prophylaxis was continued until M6. After M6, patients 

were monitored with CMV PCR monthly and treated for late 

CMV if they developed recurrent DNAemia. Fisher’s exact test 

was used. No organs were donated from prisoners. Results 

Fifty-six OLT recipients (D+ and/or R+) were monitored to 12 

months post-OLT. 47/56(84%) had a reactive QFN-CMV by 

M6. 24/56(43%) received prophylaxis or treatment before M6 

(either high-risk or developed early CMV DNAemia). 18/24 

had developed a reactive QFN-CMV by M6, of whom 78% 

had already developed a reactive QFN-CMV by 4 months. 

3/56(5%) of patients developed late CMV (median 251 days 

post-OLT). 1 asymptomatic patient commenced treatment with 

viral load 1100 copies/mL, while two developed CMV disease 

with very high viral loads (79,970 and 276,380 copies/mL). 

All 3 had a n.r. M6 QFN-CMV, suggesting they were high risk 

of late CMV following cessation of therapy. A M6 n.r. QFN- 

CMV was significantly associated with late CMV p=0.003, 

with an OR of 51.2 (sens 1.00, spec 0.89, PPV = 0.33, NPV = 

1.00). Conclusion Only 5% of recipients developed late CMV, 

but 2 of 3 suffered CMV disease. A n.r. M6 QFN-CMV is significantly 

associated with risk of late CMV, and QFN-CMV-ve 

patients likely require either extension of antiviral treatment 

or more regular monitoring. Conversely, M6 QFN-CMV has 

a high NPV, suggesting patients with robust ex-vivo immune 

response towards CMV at M6 can cease further monitoring. 

We also show that 78% of patients on antivirals may have 

protective immunity towards CMV before 4 months post-OLT, 

and antiviral prophylaxis could be discontinued early in this 

group with potential to reduce both drug side-effects and costs. 

Disclosures: 

Siddharth Sood - Grant/Research Support: Cellestis Ltd 

Robert M. Jones - Speaking and Teaching: Novartis, Astellas, Roche, Novartis, 

Astellas, Roche, Novartis, Astellas, Roche, Novartis, Astellas, Roche 



1226 

Everolimus is Associated with Lower Weight Gain Two 

Years Following Liver Transplantation – Results of a 

Randomized Multicenter Study 

Michael R. Charlton 1 , Mary E. Rinella 2 , Julie Heimbach 3 ; 1 Intermountain 

Medical Center, Salt Lake City, UT; 2 Northwestern University, 

Chicago, IL; 3 Mayo Clinic, Rochester, MN 

Background: The prevalence and severity of obesity increase 

following liver transplantation. Complications of obesity, including 

posttransplant metabolic syndrome, are an important cause 

of post-transplant morbidity and mortality. mTOR inhibitors have 

been shown to lead to lower body mass when compared to 

calcineurin inhibitors in animal studies. In contrast, calcineurin 

inhibitor-based immunosuppression is associated with steady 

weight gain in liver transplant recipients. Aims: To determine 

the comparative impact of mTOR inhibition on the course of 

post-transplant weight gain in humans following liver transplantation 

in the context of a randomized, controlled trial. Methods: 

Per protocol data on change in weight from baseline during 

a 24-month period were analyzed from a prospective, randomized, 

multicenter, open-label study, in which de novo liver 

transplant patients were randomized at 30 days to one of three 

immunosuppression protocols: 1) everolimus (EVR) + reduced 

tacrolimus (TAC; n = 245), or 2) TAC control (n = 243) or 3) 

TAC elimination (n = 231). Results: The treatment groups were 

well balanced in terms of age, gender, baseline weight, BMI, 

pre-transplant HCV infection and the incidence of diabetes. 

Randomization to TAC elimination was stopped prematurely 

due to a significantly higher rate of treated biopsy-proven acute 

rejection (tBPAR). Among patients who remained on treatment, 

mean increase in weight was significantly greater at month 12 

and 24 months from baseline in the TAC control arm than in the 

EVR + reduced TAC and the TAC elimination group (p=0.037- 

0.001, see table). Study medication was discontinued due to 

adverse events in 28.6% of EVR + reduced TAC and 18.2% 

of TAC control patients. Average daily prednisone doses were 

similar between treatment arms. The frequency of cardiovascular 

events and new onset diabetes mellitus was similar between 

treatment arms. Hyperlipidemia was more frequent in the EVR 

containing arms (p


1227 

Glycine is graft protective and preserves kidney function 

after liver transplantation: Data of the HEGPOL-Trial 

[ISRCTN69350312] 

Peter Schemmer 1 , Arash Nickkholgh 1 , Georgios Polychronidis 1 , 

Steffen P. Luntz 2 , Ertan Mayatepek 3 , MarkusW. Buechler 1 , Ernst 

Klar 4 ; 1 General, Abdominal and Transplant Surgery, Ruprecht- 

Karls University of Heidelberg, Heidelberg, Germany; 2 Coordination 

Center for Clinical Trials, Ruprecht-Karls University of 

Heidelberg, Heidelberg, Germany; 3 Department of General Pediatrics, 

Heinrich-Heine-University,, Düsseldorf, Germany; 4 Department 

of Surgery, University of Rostock, Rostock, Germany 

Background. Glycine, a non-essential amino acid, prevents 

tissues from several types of injury in various experimental 

models. Thus the HEGPOL-Trial was designed to assess both 

the efficacy and safety of glycine in liver transplantation (LTx). 

Methods/Materials. A total of 130 patients undergoing primary 

LTx were randomized in a two-arm placebo-controlled 

multicenter double-blinded trial. While sixty six recipients 

were treated with i.v. glycine (4.4 %; 250 mL) controls (n=64) 

received injectable water (250 mL) during the anhepatic phase. 

The same infusion was repeated once a day for the following 

7 postoperative days (POD 1-7). Primary endpoints were peak 

levels of aspartate-aminotransaminase (AST), alanine-aminotransaminase 

(ALT) and graft survival. Secondary endpoints 

were liver injury based on histology, liver perfusion, AUC of 

both AST and ALT, early graft dysfunction and cyclosporine 

A-induced nephrotoxicity. Results. Per protocol analysis has 

shown no difference between groups; however, patients with 

cut off plasma glycine values of ≥7000 mmol/L (n=29) prior to 

reperfusion showed significantly lower serum ALT levels during 

the first 25 hours after LTx. Further the eGFR was significantly 

better with glycine during the study. Patient survival was 86.2% 

compared with 72.6% in controls (p=0.08). Conclusion. 

Although the per protocol analysis could not verify the hypothesized 

effects of glycine, very high plasma concentrations of 

glycine achieved after its i.v. administration at the anhepatic 

phase and early after liver transplantation proved not only to 

be safe, but also hepatoprotective and nephroprotective. Trial 

Registration: HEGPOL; ISRCTN69350312. 

Disclosures: 

The following authors have nothing to disclose: Peter Schemmer, Arash Nickkholgh, 

Georgios Polychronidis, Steffen P. Luntz, Ertan Mayatepek, MarkusW. 

Buechler, Ernst Klar 

1228 

The Combination of Strongly (+) C4d Staining and 

High Titer Donor Specific Antibody (DSA) Suggests that 

Plasma Cell Hepatitis is a Result of Antibody-Mediated 

Rejection 

M. Isabel Fiel 1 , Sander S. Florman 2 , Thomas D. Schiano 2 ; 1 Department 

of Pathology, Icahn School of Medicine at Mount Sinai, New 

York, NY; 2 Recanati-Miller Transplant Institute, Icahn School of 

Medicine, New York, NY 

Background and Aim: Plasma cell hepatitis (PCH), is a form 

of post liver transplant (LT) allograft dysfunction. Antibody-mediated 

rejection (AMR) is being increasingly recognized as a 

cause of graft failure and is typified by high DSA titers, and 

histologically by (+) C4d immunostaining. Since PCH is felt to 

be a variant of rejection, we sought to determine whether it is 

a result of AMR. Design: PCH (characterized by >30% plasma 

cell infiltrate and severe necroinflammation) cases were identified; 

cases of acute (ACR) and chronic rejection (CR) served as 

controls. Immunostaining for C4d (ARP, Waltham MA; 1:200 

dilution) was performed on FFPE tissue using a Dako Autostainer. 

Assessment of C4d staining was performed blindly by 

a single hepatopathologist using the following scoring system: 

0=negative, 1=equivocal; 2 = 10-50% (+); 3 = moderate/ 

strong diffuse (+) in endothelial cells lining the portal venules 

and portal capillaries. Testing for DSA was performed for each 

patient; an MFI >2000 was considered significant (+). Results: 

A total of 30 (15 PCH, 15 control) cases were included; 9/15 

(60%) PCH, 4/15 (27%) controls were (+) for both C4d and 

DSA (p=0.065). A combination of strong C4d staining and 

high DSA MFI (DSA+) was associated with the diagnosis of 

PCH when compared to C4d+/DSA- cases (p=0.03). Six of 9 

(67%) PCH cases and 2/4 controls had (+) HLA Class 2 DSA 

alone; 2/9 PCH and 2/4 controls had both HLA Class 1 and 

2 (+) DSA (See Table). Four PCH patients had been receiving 

interferon therapy for recurrent HCV. One control case was 

documented to have rejection one week post-LT and had strong 

(3+) C4d staining and extremely high DSA MFI titers (class 

1=21167, class 2=22276) suggestive of true AMR. Conclusions: 

Patients with PCH have high titer DSA and strongly positive 

C4d staining of their liver allograft biopsy. Although some 

cases of ACR and CR may have high DSA MFI, C4d staining 

is less frequent and has less intensity in comparison to PCH 

cases. These findings suggest that PCH is a result of AMR. All 

HCV patients diagnosed with PCH while on interferon therapy 

showed (+) C4d staining and high DSA, suggesting that AMR 

played a role in their allograft dysfunction. Based on this data, 

the histologic diagnosis of PCH should prompt a work-up for 

AMR. 

Disclosures: 



The following authors have nothing to disclose: M. Isabel Fiel, Sander S. Florman 

1229 

The coming of age of the graft in pediatric orthotropic 

liver transplant (OLT) recipients 

Shewit Giovanni, J. Michael Millis, Andrew I. Aronsohn, John F. 

Renz, Ruba K. Azzam, Pamela Boone, Kathleen Dasgupta, Helen 

S. Te; University of Chicago Medicine, Chicago, IL 

Background: While graft survival is increasing in adults and 

children after OLT, data on long term graft function >15 years 

after OLT is scant. Aim: To describe long term graft and patient 

outcomes (>15 years) of patients who had OLT at 15 

years prior were identified from the OLT database. Patient 

demographic and laboratory data, as and radiographic and 

histologic data when available, were collected. Current immunosuppression 

and clinical outcomes were also noted. Results: 

25 patients who received OLT at a median age of 3.9 years 

(range, 0.2-16.2) were included in the study. Biliary atresia 

was the indication for OLT in 48% of cases, followed by inborn 

errors of metabolism in 16%. 22 (88%) patients received cadaveric 

grafts, of which 16 were whole and 6 were partial. The 

median duration of follow-up was 21.4 yrs (range, 15.1-28.1). 

Current immunosuppression were tacrolimus-based in 60%, 

cyclosporine-based in 12%, sirolimus alone in 8%, prednisone 

alone in 4%, and none in 16%. The latest laboratory tests are 

summarized in Table 1. 68% of patients had abnormal liver 

tests, 47% of which was cholestastic, 18% hepatocellular and


the rest was a mixed pattern. Radiologic cholangiopathy was 

seen in 9 (36%) of patients. Of 17 patients who had liver biopsies, 

chronic rejection was seen in 6 and nonspecific hepatitis 

in 4; stage 3-4 fibrosis was present in 7 patients. A total of 

76% of patients had either liver test, radiographic, or histologic 

abnormalities of the graft. There were 4 graft failures, leading 

to death in 2 patients and a repeat OLT in 1, while one is 

awaiting repeat OLT. Another patient died from sepsis related 

to hepatic abscesses. Conclusions: Liver grafts in children continue 

to function >15 years later in adulthood, but abnormal 

liver tests, radiographic or histologic findings are present in 

majority of these grafts. Chronic rejection was common, and 

graft failure that required repeat OLT or led to death occurred 

in a few patients. The presence of donor-specific antibodies in 

the patients with graft abnormalities will be investigated further. 

Disclosures: 

J. Michael Millis - Advisory Committees or Review Panels: Vital Therapies; Board 

Membership: Vital Therapies 



The following authors have nothing to disclose: Shewit Giovanni, Andrew I. 

Aronsohn, John F. Renz, Ruba K. Azzam, Pamela Boone, Kathleen Dasgupta 

1230 

Lack of effect of CMV reactivation in the outcome of 

HCV-infected liver transplant(LT)patients 

Victoria Aguilera 1,2 , Tommaso Di Maira 1 , Isabel Conde 3 , Angel 

Rubin 1,2 , Carmen Vinaixa 1,2 , Salvador Benlloch 1,2 , Martin Prieto 

1,2 , Marina Berenguer 4,2 ; 1 Hepatology Section, Hospital Universitari 

La Fe, Valencia, Spain; 2 CIBERehd, Instituto de Salud Carlos 

III, Grant PI13/01770, Valencia, Spain; 3 Hepatologia, Instituto 

de Investigación Sanitaria Hospital Universitari La Fe, Valencia, 

Spain; 4 Facultad de Medicina, Universidad de Valencia, Valencia, 

Spain 

Background: The role of CMV reactivation in recurrent hepatitis 

C after LT is controversial. Other aspects such as development of 

infections, diabetes mellitus (DM), rejection and cardio-vascular 

events (CVE) have also been associated with CMV reactivation 

post-LT. Aims: To evaluate if CMV reactivation is associated 

with (1) aggressive recurrent hepatitis C and (2) post-LT infections, 

rejection, DM and CVE. Patients and methods: Patients LT 

from Jan 2011 to Dec 2013 due to HCV-cirrhosis with HCV-positive 

viremia. CMV Reactivation was defined as CMV viral 

load(VL) >400c/ml. CMV VL was determined in a clinical basis 

during the first 12 months after LT. Outcome variables were: 

(i) aggressive recurrent hepatitis C (defined as F>1, F=0 with 

moderate inflammatory activity or cholestatic hepatitis), acute 

rejection (Banff criteria), infections, de novo DM post-LT and 

CVE. Demographics, donor, immunosuppression(IS), surgical, 

hematological, biochemical and virological related variables 

were also recorded. Results: 77 patients (mean age:56yr (25- 

68), 67% men, 46% HCC) were included. Donor and recipient 

CMV serologies were as follows: D+/R+:n=61,79%; D -/ 

R+:n=6,8%; D+/R-:4, 5%; D-/R-:n=6, 8%. CMV prophylaxis 

(during 90 days) was administered in 7(9%) patients. Reasons 

for prophylaxis were: CMV-D +/R - n=3, Methylprednisolone 

boluses or basiliximab use, n=4. CMV reactivation occurred in 

26 patients(33%), 24(34%) without prophylaxis and 2(28%) 

with prophylaxis (p=ns). Time to reactivation was 138 vs 45 

days in those with and without prophylaxis (P=.026). Reactivation 

led to anti-CMV treatment in 16(61%) pts. Severe recurrent 

hepatitis C occurred in 55 pts (71%), 9 (12%) pts developed 

rejection, 39 pts (51%) infections, 15(20%) de novo DM and 

6 (8%) a CVE. Nor CMV reactivation nor CMV treatment were 

associated with aggressive recurrent hepatitis C, rejections, 

infections, de novo DM or CVE. Graft and patient survival were 

not related either with CMV reactivation. There was a trend 

towards a potential protection effect of sirulimus-SRL- based IS 

and CMV reactivation (20% in SRL group vs 39% in non SRL 

group, p=0.1) but triple therapy or basiliximab use were not 

associated with reactivation. Conclusions: CMV reactivation 

occurs frequently after LT. About half of cases require therapy. 

In patients with prophylaxis, CMV reactivation tends to occur 

at later time points. Given the lack of association between 

CMV reactivation and post-transplant outcome, we believe it 

is unnecessary to prolong CMV prophylaxis for more than 3 

months, as suggested by some authors. mTor inhibitors seem to 

protect against CMV reactivation; larger studies are needed to 

confirm this finding.. 

Disclosures: 

The following authors have nothing to disclose: Victoria Aguilera, Tommaso Di 

Maira, Isabel Conde, Angel Rubin, Carmen Vinaixa, Salvador Benlloch, Martin 

Prieto, Marina Berenguer 

1231 

Blood Phosphatidyl ethanol identifies alcohol use 

among liver transplant recipients 

Michael F. Fleming 2 , Jenny Vue 1 , Michael R. Lucey 1 ; 1 Medicine, 

Univ. of Wisconsin School of Medicine and Public Health, Madison, 

WI; 2 Family Medicine, Northwestern University, Chicago, IL 

All patients with liver failure due to alcohol use disorder (AUD) 

who undergo liver transplantation (LT) are advised to abstain 

from alcohol. Monitoring alcohol use in AUD -LT recipients is 

hampered by lack of easily usable methods to identify alcohol 

relapses. Phosphatydylethanol (PEth), a red cell-membrane 

phospholipid that is released into circulation after alcohol use, 

accurately identifies recent alcohol use (i.e. in the prior 30 

days) in healthy subjects and AUD patients. The whole blood 

concentration of PEth varies in a dose-response fashion with 

consumption, with >8ng/ml being considered a positive test 

for recent exposure. There are no known false positive tests 

and PEth is considered 100% specific. PEth can detect daily 

drinking of > 2 drinks or intermittent binge drinking. We report 

a prospective study of blood PEth as a marker of alcohol use 

among 213 LT recipients at 2 US liver transplant centers. All 

subjects gave informed consent. The protocol was approved by 

the IRBs at each institution. Methods: The sample included 147 

men and 66 women with a mean age of 59 years. 131 had 

a history of alcohol dependence prior to LT and 82 served as 

non-alcohol dependent controls. Subjects participated in a face 

to face interview to assess recent alcohol use using a 30-day 

timeline follow-back calendar procedure and dried blood spot 

collection at study entry, and at 6 and 12 months thereafter. 

PEth was determined using an Agilent Zorbax chromatography-tandem 

mass spectrometry procedure and was performed 

by the United States Drug Testing Laboratories in Chicago. 

Results: among the available data, 49 subjects (23% of the 

total sample), including 39 AUD subjects (30% of the AUD 

group) and 10 controls (12% of the control group), had a positive 

test of >8 ng/ml, either at baseline or during the follow-up 

period. 149 patients reported no alcohol use and had a negative 

PEth assays. 32 subjects (15% of the total sample) reported 

no alcohol use and had a positive test indicating recent alcohol 

use. Of the 25 subjects who reported recent alcohol use, 11


had a positive test and 14 were negative. The 14 drinking subjects 

who were PEth-negative reported low levels of use. This 

study confirms that, despite advice to abstain, more than one 

quarter of alcohol-addicted LT recipients return to alcohol use. 

PEth identifies alcohol use in LT recipients who deny drinking. 

Liver transplant centers may want to include PEth as part of 

routine post-LT care in order to detect alcohol use in recipients 

who deny drinking and thereby provide them with early interventions, 

designed to restore sobriety. 

Disclosures: 

Michael R. Lucey - Advisory Committees or Review Panels: Galectin; Grant/ 

Research Support: Vertex, Abbvie, Gilead, Salix 

The following authors have nothing to disclose: Michael F. Fleming, Jenny Vue 

1232 

A New Score based on explant pathology allows an 

individualized prediction of HCC-recurrence after liver 

transplantation 

Charlotte E. Costentin 1 , Giuliana Amaddeo 1 , Christophe Duvoux 1 , 

Julien Calderaro 2 , Alexis Laurent 3 , Ariane Mallat 1 , Françoise 

Roudot-Thoraval 4 ; 1 Hepatology, Hôpital Henri Mondor, Creteil, 

France; 2 pathology, Henri Mondor Hospital, Creteil, France; 

3 Surgery, Henri Mondor Hospital, Creteil, France; 4 Public Health, 

Henri Mondor Hospital, Creteil, France 

Aim: After liver transplantation (LT) for hepatocellular carcinoma 

(HCC), a standardized prediction of recurrence would 

valuable in order to define patients who might benefit from post 

LT adjuvant therapy or early changes in immunosuppressive 

regimens. We recently compared the accuracy of 4 explantbased 

models for prediction of recurrence (EASL 2015). In 

this study, the “Up to seven model”, using 2 variables (size 

of largest nodule and number of nodules) appeared to be the 

best tool to identify patients with a high risk of recurrence at 

5 years post LT. The aim of this study was to design a new 

explant-based model including vascular invasion and tumor differentiation 

as predictors of recurrence and to test its accuracy 

against the “Up to seven” model. Methods: Following uni- and 

multi-variate analysis of pathological predictors of recurrence 

in a series of 372 liver explants of patients transplanted for 

HCC between 2003 and 2005 and followed prospectively for 

5 years, we built 2 predictive models. New score 1 included 

all independent pathological predictors (number of nodules, 

size of the largest, tumor differentiation, micro/macrovascular 

invasion, tumor burden (uni or bilobar)) and ranged from 0 to 

9 points. New score 2 included all above mentioned variables 

except tumor differentiation (a variable that is not available 

when tumor is totally inactivated after bridging therapy) and 

ranged from 0 to 11 points. The accuracy of these 2 scores 

and the “Up to seven” model without vascular invasion to predict 

5 year-HCC recurrence was assessed by comparison of 

AUCs of ROC curves which were subsequently compared using 

the Hanley&McNeil method. Results: The “Up to seven model” 

identified two distinct risk groups for 5 year-recurrence of HCC 

(10.8±2.2% if score ≤7 ; 54.5±4.5% si score >7). New scores 

1 and 2 identified 4 groups of patients with different levels 

of risks of recurrence ranging from 10 to >70% at 5 years. 

AUCs were 0.7915 [0.73394 - 0.84907] for the “Up to seven 

model, 0.7881 [0.73135 - 0.8448] for New score 1 and 

0.7889 [0.7352 - 0.8427] for New score 2 (p=0.743). Tumor 

differentiation did not improve accuracy to predict 5 year-HCC 

recurrence (New score 1 vs New score 2, p=0.49). Using 

New score 2, a nomogram was built, giving the probability of 

recurrence at 1, 3 and 5 years post transplantation, according 

to the score value (fig) Conclusion: A new score using number 

of nodules, size of the largest, vascular invasion and tumor 

burden on explant has similar accuracy compared to the “Up 

to seven” model to predict 5 year-HCC recurrence but allows a 

more individualized prediction of recurrence based on patients’ 

profile of explant findings. 

Disclosures: 






Roche 

The following authors have nothing to disclose: Charlotte E. Costentin, Giuliana 

Amaddeo, Julien Calderaro, Alexis Laurent, Ariane Mallat 

1233 

Comparative analysis of renal dysfunction in recipients 

with NASH compared to controls with alcoholic liver 

disease after liver transplantation 

Cheri Ogwo 1 , Satheesh Nair 2,1 , Jason Vanatta 2,1 , Vinaya Rao 2,1 , 

Chibuzor Iwelu 1 , James Eason 2,1 , Sanjaya K. Satapathy 2,1 ; 1 Surgery, 

University of Tennessee Health Sciences Center, Memphis, 

TN; 2 Surgery, Methodist University Hospital, Memphis, TN 

BACKGROUND: Association between NAFLD/NASH and 

chronic kidney disease (CKD) has been well recognized. 

Release of pro-inflammatory mediators from steatotic liver or 

through the contribution of metabolic syndrome (Obesity, type 

2 DM, dyslipidemia, and hypertension) has been postulated 

as the etiology. AIM: We aim to assess if the predisposition 

for renal dysfunction persists after liver transplantation (LT) in 

recipients with NASH compared to controls with alcoholic liver 

disease (ALD). METHODS: Charts of 806 adult LT recipients 

(6/2006- 12/2012) reviewed; 108 recipients with NASH, 

and 122 with ALD were identified. After exclusion of SLK transplants 

(9/11), re-transplants (3/5), death ≤ 90 days (3/7); 92 

NASH recipients were compared with 99 recipients with ALD. 

eGFR before LT, at 3 months, 1 yr, and 3 yrs. were compared. 

RESULTS: NASH patients are more likely to be older (57.7 yrs. 

vs 55 yrs., p=0.01), obese (32±5.4 vs. 28.4±5.6, p=0.001), 

and diabetic (42.4% vs. 38.4%, p=0.003), and have lower 

MELD score at listing (20.63±7.32 vs. 22.08±6.12, p=0.04). 

Both groups received our standard steroid-free protocol with 

rabbit ATG induction with low-dose Tacrolimus. eGFR pre-LT in 

NASH and ALD group were comparable (76±45 vs. 78±33, 

p=0.19). A significant decline in eGFR (Figure 1) was noted 

in NASH recipients at 3 mos. (61±30 vs. 69±36, p=0.15), 1 

yr. (60±20 vs. 70±25, p=0.03), and 3 yrs. (51±17 vs.68±18, 

p=0.001). Based on eGFR groups (Gr 1: eGFR < 45, Gr II: 

eGFR ≥45-60, eGFR >60), a distinct pattern of either inferior 

recovery of renal function or decline in renal function was noted 

in NASH recipients post LT (Figure 1). Declining eGFR in the 

NASH group was more likely in diabetics (46±16 vs. 67±23, 

p=0.02), but not in non-diabetics (57±17 vs. 68±17, p=0.17) 

at 3 yrs. of follow up. CONCLUSION: LT recipients with NASH 

phenotype have an increased propensity for renal dysfunction 

compared to ALD particularly in diabetics. Early conversion to 

calcineurin inhibitor sparing agents in LT recipients with NASH 

and diabetes should be considered as a strategy to preserve 

renal function.


Disclosures: 

Satheesh Nair - Advisory Committees or Review Panels: Jansen; Grant/Research 

Support: Gilead; Speaking and Teaching: Abbvie, Valeant, BMS 

Sanjaya K. Satapathy - Advisory Committees or Review Panels: Gilead, Intercept; 

Grant/Research Support: Genfit, Gilead, Biotest 

The following authors have nothing to disclose: Cheri Ogwo, Jason Vanatta, 

Vinaya Rao, Chibuzor Iwelu, James Eason 

1234 

Long-Term Outcome of Extrahepatic Biliary Atresia into 

adult life 

Javaid Sadiq 1 , Aditi Kumar 4 , Hifsa Sohail 3 , Carla Lloyd 3 , James 

W. Ferguson 4 , Darius Mirza 2 , Khalid Sharif 1 , Gideon Hirschfield 4 , 

Deirdre A. Kelly 3 ; 1 Paediatric Hepatobiliary Transplant Surgery, 

Birmingham Children’s Hospital Birmingham UK, Birmingham, 

United Kingdom; 2 Liver Surgery, University Hospital Birmingham, 

Birmingham, United Kingdom; 3 Liver unit, Birmingham Children’s 

Hospital, Birmingham, United Kingdom; 4 Liver unit, University Hospital 

Birmingham, Birmingham, United Kingdom 

Background:Biliary Atresia(BA) is the single commonest cause 

of neonatal cholestasis leading to cirrhosis,portal hypertension 

and liver failure and is the main indication for pediatric liver 

transplant(LT). Aim:Evaluate the long-term outcome of children 

with BA transitioning to adult life Subjects & Methods:Records 

of patients of BA managed over a period of 34 years(1980- 

2014) at a single institution were retrospectively reviewed 

.Patients with more than 10 years of follow-up were included in 

the study.Data collection included demographics,age at Kasai 

Portoenterostomy(KPE),associated malformations, survival with 

native liver or post-LT,mortality, current education/work/marital/family 

status. Results: 493 BA patients were managed 

during this period(260 F & 233 M).Median age at kasai was 

53 days(range:7-183 days). 92 % had isolated BA while 8 % 

had BA polysplenia malformation syndrome.332 patients were 

included in this study (1980 – 2004). 11 patients were lost to 

follow-up. Median patient survival is 17.3 yrs(0.32- 34.6) & 

median survival with native liver is 2.25 yrs(0.07-34.6). 53 

patients(16.5%) died in pediatric care; 26 with their native 

livers & 27 after LT.135 patients(50.3%) are still in pediatric 

care(Group A).57 are surviving with their native liver(A1) 

while 78 children have been transplanted(A2).7 patients are 

awaiting transplant in Group A1.133(49.6 %) patients were 

transferred to adult services(Group B); 49 with native livers(B1) 

and 84 after LT(B2). 28 patients in group B1 had portal hypertension(PH);20 

treated with beta blockers,esophageal banding 

or shunts. 9 patients transferred to adult services with native 

liver(B1) subsequently required LT & 7 are listed for LT due to 

decompensated liver disease.6 patients in group B2 required 

retransplant. After transfer to adult care, 3 patients in Group B1 

died( one due to ruptured splenic aneurysm & 2 due to decompensated 

liver disease) while 5 patients in Group B2 died from 

post-transplant lymphoproliferative disorders(PTLD),Hepatopulmonary 

syndrome, ruptured psoas cyst and bleeding & chronic 

rejection).Out of 268 patients in this series, majority participated 

in normal school education while 32(12 %) required 

special needs support. 29 transferred went to university, 18 

obtained non-vocational qualifications and 33 joined various 

training courses. Conclusion: Improved medical and surgical 

techniques have improved the outcome and quality of life for 

patients with BA, allowing them to live into adult life, complete 

their education & function as useful members of the society 

Disclosures: 





Deirdre A. Kelly - Consulting: Sanofi Pasteur; Grant/Research Support: BMS, 

Astellas, Acitllion, MSD, Roche 

The following authors have nothing to disclose: Javaid Sadiq, Aditi Kumar, Hifsa 

Sohail, Carla Lloyd, Darius Mirza, Khalid Sharif 

1235 

Assessment of Energy Metabolism in Liver Transplant 

Recipients 

Ajay Singhvi 1 , H. Steven Sadowsky 2 , Ayelet Cohen 1 , Alysen 

Demzik 1 , Mary E. Rinella 1 , Lisa B. VanWagner 1 , Josh Levitsky 1 ; 

1 Division of Gastroenterology & Hepatology, Northwestern Memorial 

Hospital, Chicago, IL; 2 Physical Therapy & Human Movement 

Sciences, Feinberg School of Medicine, Chicago, IL 

Background: The reasons for accelerated weight gain and metabolic 

syndrome following liver transplantation (LT) are not 

well understood. We hypothesized that this may be related 

to ineffective metabolic rates compared to healthy patients, 

particularly during exercise. The purpose of this pilot study was 

to assess energy expenditure during exercise in LT recipients. 

Methods: We consented ten subjects who were transplanted for 

NASH or cryptogenic cirrhosis (>1 year post) to undergo analysis 

of body composition, resting energy expenditure (REE), and 

peak oxygen uptake (VO 2 max 

). The latter was conducted using 

a ramped-Bruce protocol. VO 2 max 

was assessed by standard 

parameters, including respiratory exchange ratio and a rating 

of perceived exertion. Predicted VO 2 

was determined using the 

Wasserman-Hansen equations and REE was determined using 

the Harris-Benedict equation. Measured VO 2 max 

values were 

then compared to data from the NHANES database for ageand 

sex-matched healthy individuals. Results: Subjects (50% 

male) were mean age 60.1±4.3 years, BMI 30.7±3.22 kg/ 

m 2 , time post-LT 5.2±2.6 years. Average percent body fat was 

22.6±8.4% in males and 28.5±2.9% in females. Mean REE 

was 97.3% of predicted REE in males and 77.9% of predicted 

REE in females. Average VO 2 max 

(mL/kg/min) in LT males was 

16.9±5.0 compared to a predicted value of 21.0±3.1 (80.4% 

of predicted VO 2 max 

, p=0.04). LT females had a mean VO 2 

max 

(mL/kg/min) of 15.1±2.2 compared to a mean predicted 

value of 20.1±1.1 (75.4% of predicted VO 2 max 

, p=0.004). 

These values were lower than those of historical healthy controls 

who had a mean VO 2 max 

(mL/kg/min) of 31 in males 

and 23 in females (Figure 1). Discussion: Our results suggest 

that LT recipients, particularly females, have lower resting and 

exercise energy expenditure compared to predicted values and 

healthy controls. This may contribute to post-LT weight gain and 

inability to lose adequate weight with diet and exercise. Further 

study is necessary to investigate the pathophysiology of this 

phenomenon, the metabolic change from pre- to post-LT, and


interventional exercise/weight loss trials tailored to LT recipients’ 

metabolic capacities. 

Disclosures: 

Josh Levitsky - Consulting: Transplant Genomics Incorporated 

The following authors have nothing to disclose: Ajay Singhvi, H. Steven Sadowsky, 

Ayelet Cohen, Alysen Demzik, Mary E. Rinella, Lisa B. VanWagner 

1236 

Retained HLA Class II Donor Specific Antibody after 

Liver Transplantation Increases Risk of Acute Cellular 

Rejection: A Case Control Study 

Mohamed Elfeki 1,3 , Mahmoud El-Bendary 3 , Petrina V. Genco 2 , 

Raghda Farag 3 , Youssef Mosaad 4 , Justin H. Nguyen 1 , Denise M. 

Harnois 1 , Surakit Pungpapong 1 ; 1 Transplant, Mayo Clinic, Jacksonville, 

FL; 2 Laboratory Medicine and Pathology, Mayo Clinic, 

Jacksonville, FL; 3 Tropical Medicine and Hepatology, Mansoura 

University, Faculty of Medicine, Mansoura, Egypt; 4 Clinical pathology 

and Immunology, Mansoura University, Faculty of Medicine, 

Mansoura, Egypt 

Background: Liver is an organ that is well-known for its immune 

tolerant capacity. Conflicting results regarding liver graft 

function and rejection risk affected by pre-transplant positive 

lymphocyte cross match (LCM) or post-transplant circulating 

donor specific antibody (DSA) have been reported according 

to individual institutions. We aimed to evaluate the clinical 

significance of positive LCM coupled with positive DSA at time 

of and following orthotopic liver transplant (OLT) on outcome 

of graft function as regards to biopsy-proven acute cellular 

rejection (BPACR) and early allograft dysfunction (EAD). EAD 

was defined as the presence of one or more of the following 

postoperative laboratory analyses reflective of liver injury and 

fuction: bilirubin ≥10mg/dl on day 7, International normalized 

ratio ≥1.6 on day 7, and alanine or aspartate aminotransferases 

〉2000 IU/L within the first 7 days. Methods: This is a 

retrospective case control study of prospectively collected data 

from 586 OLT performed at our institution between December 

2009 and December 2013. Total of 37 cases with positive 

LCM and DSA at the time of OLT were identified. Each case 

was matched to two controls (74 controls) based on diagnosis 

of liver disease, age and sex of recipients. Results: Majority of 

the cases (90%) were females and they had a higher number 

of parity when compared to female controls (mean 2.6 vs. 

1.7, p=0.003). BPACR occurred significantly more among the 

cases than the controls (51% vs. 24%, p=0.006) with an odds 

ratio (OR) of 3.28 and 95% confidence interval (CI) of 1.42- 

7.57. Higher number of the cases experienced EAD compared 

to the controls (22% vs. 9%, p=0.08) with an OR of 2.64 (95% 

CI = 0.88-7.96). Despite similar immunosuppressive regimen 

per our institutional protocol, the cases who retained HLA class 

II DSA post-OLT experienced BPACR more often than the controls 

(80% vs. 24%, p


tion and patient and graft survival. Implications for the clinical 

setting are conscientiously monitoring of the GFR in the first 

months after liver transplantation and taking action as early 

as possible to prevent worsening of the creatinine clearance. 

Disclosures: 

Faouzi Saliba - Advisory Committees or Review Panels: Novartis, Roche, Genzyme, 

Vital therapies; Grant/Research Support: Astellas; Speaking and Teaching: 

Gambro, MSD, Gilead 

Rene Adam - Grant/Research Support: Merck Serono, Roche, Sanofi aventis, 

astellas, novartis, Merck Serono, Roche, Sanofi aventis, astellas, novartis, Merck 

Serono, Roche, Sanofi aventis, astellas, novartis, Merck Serono, Roche, Sanofi 

aventis, astellas, novartis 



The following authors have nothing to disclose: Noémie Minczeles, Valérie Delvart, 

Teresa Antonini, Rodolphe Sobesky, Philippe Ichai, Audrey Coilly, Bruno 

Roche, Marc Boudon, Eric Vibert, Denis X. Castaing, Daniel Cherqui, Gilles 

Pelletier, Jean-Charles Duclos-Vallee 

1238 

Value of Magnetic Resonance Cholangiography in 

Assessment of Non-Anastomotic Biliary Strictures after 

Liver Transplantation 

A. Claire den Dulk 1 , Martin N.J.M. Wasser 2 , Francois Willemssen 

3 , Melanie A. Monraats 2 , Marianne de Vries 3 , Rivka van den 

Boom 2 , Jan Ringers 4 , Hein W. Verspaget 1 , Herold J. Metselaar 5 , 

Bart van Hoek 1 ; 1 Gastroenterology and Hepatology, Leiden University 

Medical Center, Leiden, Netherlands; 2 Radiology, Leiden 

University Medical Center, Leiden, Netherlands; 3 Radiology, Erasmus 

Medical Center, Rotterdam, Netherlands; 4 Transplant Surgery, 

Leiden University Medical Center, Leiden, Netherlands; 5 Gastroenterology 

and Hepatology, Erasmus Medical Center, Rotterdam, 

Netherlands 

Background Non-anastomotic biliary strictures (NAS) remain 

a frequent complication after orthotopic liver transplantation 

(OLT). The aim of this study was to evaluate whether Magnetic 

Resonance Cholangiography (MRCP) could be used to detect 

or exclude NAS and grade the severity of biliary strictures. 

Methods In total, 58 patients after OLT from two transplantation 

centres in whom endoscopic (ERCP) or percutaneous transhepatic 

cholangiography (PTC) and MRCP were performed 

within less than 6 months were included in the study. Of these 

patients, 41 had NAS and 17 were without NAS based on 

ERCP or PTC and follow-up. Four radiologists – two in each 

center – used an adapted validated classification –termed 

Leiden Biliary Stricture Classification (LBSC)– to evaluate the 

MRCP independently and assess NAS severity on a scale from 

0 to 3 points in three hepatobiliary regions.(Fig 1) A maximum 

of 15 points could be obtained. Interobserver agreement of the 

severity score and intra-observer agreement between ERCP/ 

PTC and MRCP for each region was calculated with the kappa 

(κ) statistic. Results Optimal cut-off value of the LBSC to detect 

the presence of NAS with MRCP was calculated at ≥ 3 points 

for all readers. Applying this cut-off, sensitivity for each reader 

was >90%, with a corresponding specificity of 50-82%, positive 

predictive value (PPV) of 86-91%, and negative predictive 

value (NPV) of 80-100%. When the cut-off value was applied 

to the radiologists’ mean scores sensitivity was 98%, specificity 

65%, PPV 87% and NPV 92%. MRCP performance was better 

in evaluation of the intrahepatic bile ducts than of the extrahepatic 

bile ducts. The additional value of MRCP for grading 

severity (κ= 0.2 – 0.7) and localizing NAS (κ= 0.2 – 0.9) was 

limited. Conclusion MRCP is a reliable tool to detect or exclude 

non-anastomotic biliary strictures after OLT. MRCP cannot be 

used to reliably grade the severity of these strictures. 

Disclosures: 



The following authors have nothing to disclose: A. Claire den Dulk, Martin N.J.M. 

Wasser, Francois Willemssen, Melanie A. Monraats, Marianne de Vries, Rivka 

van den Boom, Jan Ringers, Hein W. Verspaget, Herold J. Metselaar 

1239 

Are cirrhotic patients awaiting liver transplantation protected 

against vaccine preventable diseases? 

Mazzola Alessandra 1,2 , Margherita Tran Minh 1,3 , Raluca Pais 1 , 

Pascal Lebray 1 , Denis Bernard 4 , Claire Goumard 5 , Yvon Calmus 1 , 

Filomena Conti 1 ; 1 Unité Médicale de Transplantation Hépatique, 

APHP, Hôpital Pitié Salpétrière, Paris, France; 2 Sezione di Gastroenterologia, 

University of Palermo, Palermo, Italy; 3 Clinical 

and experimental Medecine, Università del Piemonte Orientale, 

Novara, Italy; 4 Service d’Anesthésie Réanimation, APHP, Hôpital 

Pitié Salpétrière, Paris, France; 5 Sevice de Chirurgie Hépatobiliaire 

et de Transplantation, APHP, Hôpital Pitié Salpétrière, Paris, 

France 

Background: Cirrhotic patients are spontaneously immunocompromised 

and are at increased risk of infection with higher 

morbidity and mortality. Vaccination may reduce the mortality 

related to infectious complications in those patients. Only 

few data regarding protection against vaccine preventable 

diseases and the results of vaccination are available in cirrhotic 

patients awaiting liver transplantation (LT). Aims: The aims of 

this prospective non-randomized study were to prospectively 

assess the prevalence of hepatitis A, B and varicella viruses 

(HAV,HBV,VZV) serological markers and to the evaluate the 

immunization status in a cohort of cirrhotic patient awaiting LT 

to determine the need of vaccination in this cohort. Patients and 

methods: All the cirrhotic patients registered on the LT waiting 

list in a single center were questioned about the prevalence of 

previous diseases and vaccinations by a questionnaire. The 

enzyme linked immunosorbent assay (ELISA) method was used 

to assess the presence of HBsAg, anti-HBc, anti-HBs, anti-HAV 

and anti-VZV antibodies. The positivity of serology was defined 

as: HBsAg ≥1 mUI/mL, anti-HBs ≥10 mUI/L, anti-HBc≥1 mUI/L, 

anti-HAV(IgG)≥1mUI/mL and anti-VZV(IgG)≥165 mUI/mL for 

HBV, HAV and VZV viruses, respectively. Results: 201 patients 

(male: 74%, mean age: 54±11years) have been evaluated. 

The indication for LT was HCV-related cirrhosis in 42.4%, HBV 

cirrhosis in 9%, alcoholic cirrhosis in 43.1%, NASH in 6.1%, 

hepatocellular carcinoma in 41.9%, and other indications in 

21.2%. Forty-five percent were class A, 33.2% B and 18.9% 

C according to Child-Pugh score; median MELD score was 12 

(6-33). Only 2.5% of patients had a vaccination book, 50% 

were anaware of having or having not a pasthistory of HBV, 

HAV or VZV infection. Twenty-nine percent, 6.7% and 4.0%, 

respectively, reported a vaccination against HBV, HAV and 

VZV. Seroprevalence was 11.1% for HBsAg, 32.2% for anti-


HBc, 34% for anti-HBs, 19.8% for anti-HBc-/anti-HBs+, 79.4% 

for anti-HAV(IgG), and 91.7% for anti-VZV(IgG). Conclusion: 

Despite vaccination recommendations in cirrhotic patients 

awaiting LT, only few patients are informed and vaccinated 

while on the LT waiting list in a large Parisian center. Serological 

protection for VZV and HAV was high (91.7 and 79.4 

respectively), but it was low for HBV (34%), demonstrating the 

urgent need of vaccination management in this population. 

Disclosures: 

Pascal Lebray - Grant/Research Support: Merck, astellas, Biotest, BMS; Speaking 

and Teaching: Janssen, MSD, Gilead 

The following authors have nothing to disclose: Mazzola Alessandra, Margherita 

Tran Minh, Raluca Pais, Denis Bernard, Claire Goumard, Yvon Calmus, Filomena 

Conti 

1240 

Increase in Red Cell Distribution Width Post Liver Transplant 

is Associated with Patient Mortality 

Thure Caire 1 , Kirbylee K. Nelson 2 , R. Todd Stravitz 4 , Ambuj 

Kumar 2 , Nyingi M. Kemmer 3 ; 1 Gastroenterology, USF, Brandon, 

FL; 2 Internal Medicine, USF, Tampa, FL; 3 Hepatology, Tampa General 

Medical Group, Tampa, FL; 4 VCU, Richmond, VA 

Background: Following liver transplantation (LT), patients are at 

significant risk of mortality, however, prognostication following 

LT is limited. The red cell distribution width (RDW) is a measure 

of anisocytosis, and has been associated with morbidity and 

mortality in several chronic diseases associated with chronic 

systemic inflammation. The association of changes in RDW with 

mortality in patients following transplant surgery is unknown. 

As a potential surrogate for immunopathology, we hypothesize 

an increase in RDW may be positively correlated with mortality 

risk, therefore, the aim of this study is to determine the association 

between changes in RDW following LT and patient mortality. 

Methods: We performed a retrospective cohort study of all 

LT recipients performed at our center from Jan 1 2012 – Dec 

31 2012. The data collected included demographics (age, 

gender, ethnicity/race, indication for LT), pre-transplant RDW 

lab data (MELD score, hematologic parameters including MCV 

and RDW, Liver panel, renal profile) with a 24 month follow-up 

period including lab data at 6 months post-LT, and mortality 2 

years post-LT. We retrospectively analyzed data for associations 

with survival. Our primary end point was association of 

change in RDW (pre-LT to 6 months post-LT) with post-transplant 

mortality at 2 years. Statistical analysis was done using chisquare, 

fischer exact test, regression analysis. A p-value 30kg/m 2 (OR 1.089, p = 0.048), 

use of rapamycin (OR 9.898 p < 0.001), prior intra-arterial 

HCC therapy (OR 3.761, p < 0.001), take back surgery (OR 

9.038, p < 0.001), and cold ischemic time (OR 1, p = 0.209) 

were considered significant. On multivariate analysis, only biologic 

MELD at LT (OR 0.896, p = 0.028), BMI>30kg/m 2 (OR 

1.122, p = 0.043), take back within 1 st month (OR 14.559, 

p < 0.001), and rapamycin use (OR 12.450, p < 0.001) 

remained significant. Pre-LT intra-arterial therapy was not a 

significant predictor on multivariate analysis (OR 2.546, p = 

0.140). Conclusion: Pre-transplant intra-arterial HCC therapies 

to downstage into or maintain potential LT recipients within 

Milan criteria are not associated with increased prevalence of 

HAT post-LT. With respect to pre-LT HCC intra-arterial treatment 

factors, no increased risk for HAT is conferred with modality 

(chemo vs. radio-embolization), number of sessions, or number 

of HCC foci. 

Disclosures: 

Anil B. Seetharam - Advisory Committees or Review Panels: Bristol Meyers 

Squibb; Speaking and Teaching: Gilead, Janssen, Merck, Bayer 

The following authors have nothing to disclose: Mark Pedersen, Meera Ramanathan, 

Myunghan Choi


1242 

The Association of Pre-Transplant Sarcopenia and Cirrhotic 

Cardiomyopathy with Postoperative Complications 

after Liver Transplant. 

Tripti Gupta 1,2 , Ryan M. Durel 1 , Qingyang Luo 1 , Daniel Devun 1 , 

John Seal 1 , Emily Ahmed 1 , David S. Bruce 1 , Trevor W. Reichman 1 , 

Shobha Joshi 1 , Natalie H. Bzowej 1 , Nigel Girgrah 1 , Humberto E. 

Bohorquez 1 , Ari J. Cohen 1 , Ian C. Carmody 1 , George E. Loss 1 , 

George Therapondos 1 ; 1 Ochsner Clinic Foundation, New Orleans, 

LA; 2 University of Queensland, Brisbane, QLD, Australia 

Background: Sarcopenia in patients listed for liver transplantation 

(LT) is associated with increased death on the wait list 

and may affect post-operative recovery. These patients also 

have cirrhotic cardiomyopathy but its effect on post-operative 

outcomes remains unclear. Aims: To describe the prevalence of 

sarcopenia and cirrhotic cardiomyopathy in a cohort of adult 

patients with cirrhosis who are waitlisted for LT. To explore a 

correlation between these two conditions. To investigate their 

effect on post-operative outcomes. Methods: This is a retrospective 

study of 327 cirrhotic patients who underwent LT at 

Ochsner Medical Center between Jan 2012-Dec 2013. Pre-LT 

data such as demographics, clinical characteristics, etiology 

of liver disease, MELD score, 2D echocardiographic data (Left 

ventricular volume index- LAVI), E/A ratio, QTc interval were 

collected. CT abdomen/pelvis was used to calculate psoas 

muscle cross-sectional area (at L4 level). Post-LT data were also 

collected- no of days in hospital (first admission), no of ICU 

admissions, no of infections and creatinine level. Results: The 

M:F ratio was 68/32 and the mean age was 55 yrs. The mean 

MELD score was 23.5. Data for the first postoperative year 

were collected. The MELD scores and other demographics as 

well as 1-yr patient and graft survivals in the sarcopenia vs 

non-sarcopenia groups were similar. Cardiac assessment: 36% 

of our cohort had an abnormal LAVI. Sarcopenia: 19.5% of 

patients were sarcopenic. There was no correlation between 

the presence of sarcopenia and any of the abnormal cardiac 

measurements. 12.5% of patients with sarcopenia were discharged 

to a rehab facility vs only 6.8% of patients without 

sarcopenia. See Table for other post-LT outcomes. Summary: 

There was no correlation between the presence of sarcopenia 

and cirrhotic cardiomyopathy. Sarcopenia was shown to 

be associated with prolonged hospital stay, more episodes of 

infection and was more likely to lead to a discharge to a rehabilitation 

facility rather than home. The presence of cirrhotic 

cardiomyopathy pre-LT was associated with worse renal function 

at 1 year post LT. Conclusions: Sarcopenia is associated 

with increased post-LT morbidity and increased health care 

utilization while cirrhotic cardiomyopathy is associated with 

post-LT renal impairment. Although both conditions are seen 

in patients with cirrhosis, they do not appear to be associated 

with each other. 

Post-LT outocmes 

Disclosures: 

Shobha Joshi - Grant/Research Support: Salix, Eisai; Speaking and Teaching: 

Merck, Gilead, Bristol-Myers Squibb 

Natalie H. Bzowej - Grant/Research Support: Gilead Sciences, Ocera Therapeutics 

Nigel Girgrah - Speaking and Teaching: Merck, Bayer, Vertex, Gilead, Merck, 

Bayer, Vertex, Gilead, Merck, Bayer, Vertex, Gilead, Merck, Bayer, Vertex, 

Gilead 

The following authors have nothing to disclose: Tripti Gupta, Ryan M. Durel, 

Qingyang Luo, Daniel Devun, John Seal, Emily Ahmed, David S. Bruce, Trevor 

W. Reichman, Humberto E. Bohorquez, Ari J. Cohen, Ian C. Carmody, George 

E. Loss, George Therapondos 

1243 

Outcomes Post-Liver Transplant for Primary Sclerosing 

Cholangitis: Colitis Activity and Malignancy 

Astrid-Jane Greenup 1 , Gokulan Pavendranathan 1,2 , Mathew J. 

Keegan 1 , Sean Griffin 1 , David Bowen 1,4 , Warwick Selby 1,3 , Nicholas 

A. Shackel 1,4 , Simone I. Strasser 1,3 , Geoff McCaughan 1,4 , 

David J. Koorey 1,3 ; 1 A W Morrow GI and Liver Centre, Royal 

Prince Alfred Hospital, Sydney, NSW, Australia; 2 University of 

New South Wales, Sydney, NSW, Australia; 3 University of Sydney, 

Sydney, NSW, Australia; 4 Centenary Institute of Cancer Medicine 

& Cell Biology, Sydney, NSW, Australia 

Background The reported risk of colorectal cancer(CRC) in 

patients transplanted for primary sclerosing cholangitis(PSC) 

is higher than in other transplant indications and non-transplanted 

patients with PSC.Yearly endoscopic surveillance is 

recommended.Furthermore, inflammatory bowel disease(IBD) 

activity varies despite immunosuppressive therapy. Aims The 

aims of this retrospective observational study were to examine 

IBD colitis activity and rates of colonic dysplasia and cancer 

in patients post-liver transplantation for PSC. Methods Data 

for patients undergoing liver transplantation for PSC at Royal 

Prince Alfred Hospital from January 1986 to December 2014 

was collected and analysed. Results One hundred and fourteen 

patients were transplanted for PSC during this 29-year period. 

Thirty-one patients were excluded for less than 2 years of follow 

up post-transplant (deceased(n=20;19 prior to 2008) or transplantation 

since January 2013(n=11)).Of the remaining 83 

patients,14 patients had a pre-transplant colectomy,care of 4 

patients was transferred to another interstate transplant centre 

and 3 had limited available medical records.Eighteen of 62 

patients did not have IBD pre-transplant,of whom 1 required 

a colectomy(for sporadic CRC) and 4 subsequently developed 

IBD.Accordingly,post-transplant IBD outcomes were available 

for 48 patients with mean age 58 years,male to female ratio 

2:1 and diagnosis of ulcerative colitis in 28 and Crohn’s 

disease in 20.Mean follow-up was 11 years.Forty-seven of 

48 had had at least one colonoscopy;median interval 1.74 

years.Colitis was histologically active in 68%(n=32) post-transplant,including 

22 patients with quiescent disease preceding 

transplant.Three patients received tumour necrosis factor 

antagonist therapy,inducing remission in two whilst the other 

required colectomy.A further four patients needed colectomy 

for refractory colitis.In the remaining 15,colitis was quiescent 

post-transplant.Six patients had a total or hemi colectomy for 

the indications of CRC(3),invisible dysplasia(1),endoscopically 

unresectable polypoid dysplasia(1) and caecal volvulus(1).An 

additional 4 patients had a history of invisible dysplasia and 

4 endoscopically resectable polypoid dysplasia. Conclusions 

Colitis has been active in 68% of patients post-transplant for 

PSC.Dysplasia and/or CRC have been detected in 14 patients. 

Colectomy has been necessary in 11 patients overall.These 

findings emphasise the need for aggressive surveillance and 

colitis management post-transplantation, including vigilance 

for de novo IBD development.Median colonoscopic interval 

of 1.74 years remains longer than recommended and is an 

opportunity for quality improvement. 

Disclosures: 

Simone I. Strasser - Advisory Committees or Review Panels: Janssen, AbbVie, 

Roche Products Australia, MSD, Bristol-Myers Squibb, Gilead, Norgine, Bayer 

Healthcare; Speaking and Teaching: Bayer Healthcare, Bristol-Myers Squibb, 

MSD, Roche Products Australia, Gilead, Janssen, Abbvie



The following authors have nothing to disclose: Astrid-Jane Greenup, Gokulan 

Pavendranathan, Mathew J. Keegan, Sean Griffin, David Bowen, Warwick 

Selby, Nicholas A. Shackel, David J. Koorey 

1244 

The Gap Between Assessed Physical Capacity And 

Actual Activity In Advanced Cirrhosis 

Michael A. Dunn 1,2 , Deborah A. Josbeno 3 , Amy R. Schmotzer 1 , 

Doug Landsittel 1,2 , Anthony Delitto 3 ; 1 Medicine, University of Pittsburgh, 

Pittsburgh, PA; 2 Thomas E Starzl Transplantation Institute, 

University of Pittsburgh, Pittsburgh, PA; 3 Physical Therapy, University 

of Pittsburgh, Pittsburgh, PA 

Background: Muscle wasting in cirrhosis is a major cause of 

deterioration, transplant waitlist removals and deaths. It is 

associated with deconditioning and impaired physical performance, 

and becomes apparent as anatomic sarcopenia on 

imaging. Physical inactivity is a known driver of muscle wasting 

in chronic diseases. We instruct waitlisted patients to maintain 

regular activity and mandate the assessment of physical performance 

capacity for listing. However, very little is known about 

whether such assessments predict or reflect the extent of physical 

activity that patients actually perform to sustain health while 

awaiting transplantation. Methods: We asked 43 waitlisted 

patients to self-assess their ability to perform ordinary physical 

tasks using the Rosow-Breslau survey. We also recorded their 

Karnofsky physical capacity assessments from UNOS listing 

records. We then measured actual physical activity using a 

wearable armband with accelerometer and thermal sensing to 

record (a) percentage of waking time spent in sedentary (0-1.5 

METs), light (1.5-3.0 METS) and moderate-to-vigorous (>3.0 

METS) activity, (b) daily steps, and (c) daily energy expenditure 

in kcal. Results: We were able to acquire analyzable 

data for a 4 to 7 day wear period in 30 of the 43 patients 

(21 men, 9 women; age 57.5 ± 8.9; MELD 17.2 ± 6.2). Their 

measured levels of physical activity were among the lowest 

reported in chronic disease, comparable with that of COPD 

patients using oxygen or chronic renal failure patients on dialysis. 

Their percentages of waking hours spent in sedentary, 

light, and moderate-to-vigorous activity were 80.8 ± 14.7%, 

16.6 ± 13.0%, and 2.5 ± 2.7%, respectively. Compared with 

a range of 7,000-13,000 steps/day in healthy adults, their 

mean steps/day were 3132 ± 2022, range 552-8414. Both 

the activity percentage and step data were typical of other 

severely inactive populations. Mean daily energy expenditure 

was 2287 ± 468 kcal, as expected for their age range. Neither 

the self-assessed Rosow-Breslau scores (mean 2.5 ± 0.8, 

maximum 3.0) nor the provider-generated Karnofsky scores 

(mean 79.7 ± 11.3, maximum 100) indicated major impairment 

or showed a significant correlation with patients’ actual 

performance measured either by activity percentage or daily 

steps. Conclusion: Physical activity in cirrhotic patients is highly 

sedentary. Self and provider assessments of capacity do not 

correlate with actual performance. The extent to which a potential 

gap between assessed capacity and performance may be 

favorably modified by motivational, exercise, and nutritional 

interventions to improve activity and ameliorate muscle wasting 

is unclear, and merits further study. 

Disclosures: 

The following authors have nothing to disclose: Michael A. Dunn, Deborah A. 

Josbeno, Amy R. Schmotzer, Doug Landsittel, Anthony Delitto 

1245 

Liver Transplantation in Older Recipients 

Denise M. Harnois, Kaitlyn R. Musto, Bhupendra Rawal, Justin H. 

Nguyen; Mayo Clinic, Jacksonville, FL 

Introduction: The role of liver transplantation (LT) in elderly 

recipients (≥70 years) is controversial. Over the last decade, 

longevity has improved so that individuals who reach 70 years 

of age can expect an additional 15 years of life (http://cdc. 

gov/NCHS/products/nvsr.htm#vol58/). To address this issue, 

we review our experience of LT at a single large volume center 

to determine if LT is a reasonable option for older recipients to 

reclaim their expected lifespan if liver failure can be overcome. 

Methods: Retrospective analysis of 2524 LT recipients from 

1998 to 2013 was completed. Recipients of primary, whole, 

non-DCD, liver alone transplants, non-CCCA, and recipient 

age ≥18 were included. Clinically available variables including 

patient and graft survival along with donor and recipient 

characteristics were used. Results: Median recipient age was 

56 years (Range: 18-80) with a total of 1853 patients that 

included 129 older (≥70 years), 1482 middle-aged (46-69 

years), and 242 young (18-45 years) LT recipients. 1221 (66%) 

recipients were male. Median raw MELD was 16 (Range: 6-51) 

with median of 15 (Range: 6-43) for elderly. 40% had history 

of HCV and 22% with HCC. Post-LT median LOS was 7 days 

(Range: 0-446) and 8 days (Range: 0-105) for older recipients. 

933 (50.4%) of all recipients were sent to ICU post-LT, 65 

(50.4%) of older patients. Overall patient survival post-LT was 

95%. The 5-year patient survival was 85.1% (graft 73.5%) in 

young, 79.4% (graft 74.8%) in middle-aged, and 72.7% (graft 

69.0%) in older recipients. The 10-year survival was 72.1% 

(graft 59.5%) in young, 69.1% (graft 65.1%) in middle-aged, 

and 51.2% (graft 48.5%) in older recipients. Conclusion: LT 

offers the elderly a chance to reclaim their expected lifespan as 

they regain their liver function. Moreover, mortality, graft loss, 

LOS and requirement for ICU in older recipients are similar to 

those of young recipients. Therefore, LT in older recipient can 

be successful and should be considered in selected patients. 

Disclosures: 

The following authors have nothing to disclose: Denise M. Harnois, Kaitlyn R. 

Musto, Bhupendra Rawal, Justin H. Nguyen


1246 

IgG4 status in explanted livers may affect the outcome 

of Primary sclerosing cholangitis (PSC) after liver transplantation 

Mansour G. Alghanem 1 , Bandar Al-Judaibi 2 , Paul Marotta 2 , Subrata 

Chakrabari 3 , Chaturika Herath 3 ; 1 mansour alghanem, university 

of western ontario, London, ON, Canada; 2 Hepatology, 

Western university, London, ON, Canada; 3 Western university, 

London, ON, Canada 

BACKGROUND: The outcome of Primary Sclerosing Cholangitis 

(PSC) after liver transplantation can be affected by recurrent 

PSC (rPSC) and subsequent graft failure. IgG4 related sclerosing 

disease is a recent entity that has a similar morphological 

appearance to PSC, making the distinction difficult. However, 

IgG4 related sclerosing cholangitis has an excellent prognosis 

since, it is steroid sensitive, but the impact of IgG4 on rPSC 

after liver transplant is still unknown. AIM: To determine the 

association between IgG4 immunochemical staining in liver 

explants and recurrence of primary sclerosing cholangitis postliver 

transpantation METHODS: All adult patients who underwent 

liver transplantation for PSC,from 1990 to 2014, were 

identified. Clinical inforamtion and immunochemical staining 

were obtained among the patietns with PSC recurrence postliver 

transplantation. IgG4 immunochemical staining was perforemed 

on the porta-hepatis region. Immunochemical staining 

is considered to be positive if the score was >5 cells/HPF. 

RESULTS: A total of eighty patients fulfilled the criteria for the 

study. IgG4 staining was positive in 21/80 (26%) subjects 

(>5cells/HPF). Median time for follow-up in IgG4 positive 

group was 99.6 months compared to 152.6 months in the 

IgG4 negative There were more episodes of rPSC in IgG4 

positive group (P


patients had ascites, 14 had encephalopathy, and 10 had 

varices. The accelerometer data showed that liver transplant 

candidates performed remarkably little daily physical activity 

(Table). There was no correlation between MELD and physical 

activity. Additionally, physical activity was not different 

based on the presence of ascites, hepatic encephalopathy, 

esophageal varices, or etiology of liver disease. Mean moderate-vigorous 

physical activity was correlated with right (r=0.51, 

p=0.017) and left (r=0.63, p=0.002) hand-grip strength. 

There was no correlation between physical activity and six-minute 

walk test distance. CONCLUSION: Liver transplant candidates 

with a relatively low mean MELD score were extremely 

sedentary, averaging only 5 minutes/day of physical activity. 

Moderate-vigorous activity was associated with greater grip 

strength. The lack of physical activity and large amount of time 

spent sedentary offer a potential for intervention to improve 

fitness in liver transplant candidates. 

76%, P=0.32, HR [95% CI]: 0.85 [0.60-1.2]). However, at 

MELD


Comparing centers performing transplants for alcoholic hepatitis 

to centers not performing such transplants 

LT: Liver transplantation; AH: Alcoholic hepatitis; NRS: Non-response 

to steroids 

Disclosures: 


The following authors have nothing to disclose: Mohsen Hasanin, Derek Dubay, 

Thomas D. Schiano, Ashwani Singal 

1251 

15 first-days over-immunosuppression (IS) is NOT associated 

with immune-outcome measures post-liver transplantation 

(LT) 

Tommaso Di Maira, Victoria Aguilera, Angel Rubin, Carmen 

Vinaixa, Maria Garcia, Salvador Benlloch, Eva Montalva, Angel 

Moya, Fernando San Juan, Rafae Lopez-Andujar, Martin Prieto, 

Marina Berenguer; La Fe University Hospital, Valencia, Spain 

Body: Background & Aims: A recent study has described a 

strong association between early post liver transplantation (LT) 

(IS) and outcome. We aimed to confirm these findings in a 

cohort with long-term follow up. Methods: Tac and CsA trough 

levels obtained during the first 15 postLT days of patients 

transplanted between 2006-2008 were collected. High IS 

was defined by median Tac or CsA higher than 10 ng/ml or 

250 ng/ml respectively and/or the presence of a peak of Tac 

or CsA greater than 20 ng/ml and 400 ng/ml, respectively. 

Optimal IS was defined by median Tac or CsA levels between 

7-10 ng/ml or 150-250 ng/ml. Exclusion criteria were: lack of 

available data,


strategies to improve post-LT survival of patients with cirrhosis 

with history of pre-transplant acute kidney injury. 

Disclosures: 

The following authors have nothing to disclose: Paul S. Fitzmorris, Kirk B. Russ, 

Donny Kakati, Ashwani Singal 

Disclosures: 






Inc. 

The following authors have nothing to disclose: Maria Aguilar, Edward W. Holt, 

Taft Bhuket, Benny Liu, Robert J. Wong 

1253 

Renal Function Recovery and Outcomes after Liver 

Transplantation Alone among Patients with Pre-Transplant 

Acute Kidney Injury 

Paul S. Fitzmorris 1 , Kirk B. Russ 1 , Donny Kakati 1 , Ashwani Singal 

2 ; 1 Internal medicine, University of Alabama at Birmingham, 

Birmingham, AL; 2 Gastroenterology and Hepatology, University of 

Alabama at Birmingham, Birmingham, AL 

Purpose: Data on recovery of renal function recovery after liver 

transplantation (LT) alone are controversial. Methods: From an 

ongoing prospective study to define urine or serum biomarkers 

predictive of renal function recovery after LT, high risk patients 

with history of acute kidney injury (AKI) while waiting for LT 

were prospectively followed for renal function recovery at 6 mo. 

and for patient survival at 1 yr. after LT. Glomerular filtration 

rate was estimated using the modified diet in renal disease-6 

(MDRD-6) equation. Results: Of 109 patients with pre-LT AKI, 

52 successfully received LT. After excluding 3 with simultaneous 

kidney and 2 with lack of pre-transplant data, 47 (38 with 

1, 6 with 2, and 3 with 3 episodes of AKI) receiving LT alone 

were reviewed. Of these, 13 were excluded from analysis for 

lack of MDRD-6 data at 6 months after LT (n=4), not made up to 

6 months after LT (n=5), and death within 6 months of LT (n=4, 

1 each due to operative on table, sepsis, pulmonary embolism, 

and exploratory laparotomy for hemorrhage). Of remaining 34 

patients, 11 with recovery of renal function (improved MDRD-6 

by 50% compared to MDRD-6 at LT) differed from 23 who 

without recovery of renal function for MDRD-6 at LT (28±15 

vs. 60±30, p=0.002) and length of stay after LT (15±6 d vs. 

8±3 d, p=0.0002) without differences on age at LT, sex comorbidities, 

pre-existent chronic kidney disease, and number of 

AKI episodes prior to LT. Of 25 patients with MDRD-6


1255 

Role of Coronary Artery Calcium (CAC) Score in Identifying 

Patients at Risk for Cardiac Events Within the 30 

Day Postoperative Period Following Deceased Donor 


Theodore W. James 1 , Joban Vaishnav 1 , Mohammad U. Malik 2 , 

Aliaksei Pustavoitau 3 , Andrew M. Cameron 4 , Stuart D. Russell 1 , 

Ahmet Gurakar 1 ; 1 Medicine, Johns Hopkins Hospital, Baltimore, 

MD; 2 Medicine, Conemaugh Memorial Medical Center, 

Jonestown, PA; 3 Anesthesiology and Critical Care Medicine, Johns 

Hopkins Hospital, Baltimore, MD; 4 Transplant Surgery, Johns Hopkins 

Hospital, Baltimore, MD 

Objective: To determine the relationship between CAC score 

and cardiac events encountered within the 30 day period 

following deceased donor liver transplantation (LT). Methods: 

Patients with a history of hypertension, Diabetes Mellitus, smoking 

or NASH underwent CAC scoring as part of their pretransplant 

evaluation. Following IRB approval, the records of 

adult patients who underwent CAC scoring and subsequent 

deceased donor LT between 1/1/2012 and 3/1/2015, with 

a minimum of 90 day follow-up, were retrospectively reviewed. 

The discharge summaries and progress notes were searched 

for cardiac events during the index admission or 30 day postoperative 

period. Cardiac events were categorized as new 

arrhythmia, acute coronary syndrome (ACS), new heart failure 

symptoms, new valvular disease or other cardiac disease. Postoperative 

cardiac events were compared at different Agatston 

unit cutoffs via Receiver Operating Characteristic (ROC) analysis. 

Results: This study included 75 subjects; (68% males, age 

range 41-69, median age 57, median biological MELD at the 

time of transplant 20; 45 Hepatitis C, 11 Alcohol, 6 NASH, 

4 Autoimmune, 1 Primary Sclerosing Cholangitis, 1 Primary 

Biliary Cirrhosis, 7 other). Twenty recipients experienced a cardiac 

event within the 30 day postoperative period (70% males, 

median age 58, median biological MELD 20; 11 Hepatitis 

C, 3 Alcohol, 2 NASH, 1 Autoimmune, 3 other). Six patients 

experienced a new arrhythmia, 13 experienced ACS, and 

one patient experienced myopericarditis. ROC analysis demonstrated 

a maximal sensitivity and specificity for 30 day postoperative 

cardiac events (85% and 58% respectively) at 80 

Agatston units. At this cut point, the negative predictive value 

was 91% (p


were seen and evaluated for LT, 138 (59%) underwent RHC 

based on RVSP on echocardiography of >35 mm Hg. Patients 

with and without RHC did not differ for demographics, comorbidities, 

liver disease etiology, complications of liver disease 

(ascites, hydrothorax, congestive heart failure, dialysis, hepatocellular 

carcinoma) and MELD score. Of 123 patients with 

available data on pulmonary artery pressure (PAP) measurements, 

51 (41%) had pulmonary hypertension (PHT) with mean 

PAP>25 mm Hg. Patients with mean PAP>25 compared to 25 and pulmonary vascular resistance >240 

dynes) was identified in only 5 (4.7%) patients. Conclusion: 

On screening echocardiography among patients with cirrhosis 

who are candidates for LT, RVSP cut-off of 40 mm Hg is most 

optimal for further evaluation for PHT with RHC. About 5% of 

these patients have true PHT. 

Disclosures: 

The following authors have nothing to disclose: Natalie Mitchell, Fakhar U. Islam, 

Nicholas Piazza, Danisa M. Clarett, Ashwani Singal 

1258 

Pre-transplant portal vein thrombosis is an independent 

risk factor for graft loss due to hepatic artery thrombosis 

in liver transplant recipients 

Jonathan G. Stine 1 , Shawn Pelletier 4 , Timothy Schmitt 2 , Robert J. 

Porte 3 , Patrick Northup 1 ; 1 Gastroenterology & Hepatology, University 

of Virginia, Charlottesville, VA; 2 Surgery, University of Kansas, 

Kansas City, KS; 3 Surgery, University of Groningen, Groningen, 

Netherlands; 4 Surgery, University of Virginia, Charlottesville, VA 

Purpose: To examine hepatic artery thrombosis risk in liver 

transplant recipients. We hypothesize that recipients with portal 

vein thrombosis are at increased risk. Methods: Data on all 

transplants in the United States during the MELD era through 

September 2014 were obtained from UNOS. Status one, 

multi-visceral, living donor, re-transplants, pediatric recipients 

and donation after cardiac death were excluded. Recipients 

were sorted into two groups: those with HAT and those without 

and univariate comparisons were performed. Incomplete HAT 

data was excluded. Multivariable logistic regression models 

were constructed for hepatic artery thrombosis with resultant 

graft loss within 90 days of transplantation. Findings: 6,134 

recipients underwent transplantation with known hepatic artery 

thrombosis status. 662 recipients had early hepatic artery 

thrombosis; of those, 91 (13.8%) had pre-transplant portal 

vein thrombosis, versus 6.8% with portal vein thrombosis but 

no hepatic artery thrombosis (p


Multivariate Model: WL Removal Death/Too Sick 

Variables not significant: BMI, blood type, sex, dialysis, PVT, 

prior abdominal surgery 

Disclosures: 




The following authors have nothing to disclose: Jeanne-Marie Giard 

1260 

Rotational Thromboelastometry (ROTEM) versus Conventional 

Coagulation Tests during Orthotopic Liver Transplantation: 

Comparison of Intra-operative Blood Loss, 

Transfusion Requirements, and Cost 

Laura Smart 1 , Danielle T. Scharpf 2 , Nicole O’Bleness Gray 1 , 

Daniel Traetow 2 , Sylvester Black 3 , Anthony Michaels 1 , Elmahdi 

Elkhammas 3 , Robert B. Kirkpatrick 1 , Khalid Mumtaz 1 , A. James 

Hanje 1 ; 1 Gastroenterology,Hepatology, and Nutrition, The Ohio 

State University Medical Center, Columbus, OH; 2 Anesthesiology, 

The Ohio State University Medical Center, Columbus, OH; 3 Transplant 

Surgery, The Ohio State University Medical Center, Columbus, 

OH 

Purpose: Orthotopic liver transplantation (OLT) can be associated 

with significant bleeding requiring multiple blood product 

transfusions, especially in patients with severe liver dysfunction. 

Rotational thromboelastometry (ROTEM) is a point-of-care 

device that has been used successfully to monitor coagulation 

on whole blood samples during OLT. Whether it reduces 

blood loss and transfusions during OLT remains controversial. 

Methods: ROTEM or conventional coagulation tests (activated 

partial thromboplastin time (aPTT), prothrombin time (PT), international 

normalized ratio (INR), platelet count, and fibrinogen) 

were used to guide transfusion of platelets, cryoprecipitate, 

and fresh frozen plasma (FFP) during OLT. 68 consecutive 

patients were included in this non-randomized retrospective 

study. 34 historic controls had transfusions guided by conventional 

labs and 34 patients had transfusions guided by 

ROTEM during OLT over 3 years. Patient characteristics as well 

as pre- and post- transplant laboratory data were collected. 

Intra-operative blood loss, type and amount of blood products 

transfused, and cost were compared between the two groups. 

Results: The ROTEM group had significantly less intra-operative 

blood loss compared to the conventional group (2.0L vs 3.0L, 

p=0.04). The total amount of blood products transfused was 

less in the ROTEM group but did not reach statistical significance 

(14.5 units vs. 17 units, p=0.11). Patients in the ROTEM 

group received significantly less FFP (4 units vs. 6.5 units, 

p=0.02) but more cryoprecipitate (2 units vs. 1 units, p=0.04). 

Patient characteristics and pre-transplant laboratory results 

were not statistically different between groups. Post-transplant, 

both the INR and platelet count were significantly higher in the 

ROTEM group (2.0 vs. 1.7, p=0.01 and 98,000 vs. 63,000, 

p=0.002, respectively). The direct cost of blood products was 

also less in the ROTEM group compared to the conventional 

group ($103,786.09 vs $123,067.01). Conclusion: Implementation 

of a ROTEM-guided transfusion algorithm resulted 

in a reduction in intra-operative blood loss and a trend toward 

decreased total blood product usage with resultant decrease in 

cost during orthotopic liver transplantation. 

Disclosures: 

Anthony Michaels - Advisory Committees or Review Panels: Gilead, BMS, Janssen; 



The following authors have nothing to disclose: Laura Smart, Danielle T. Scharpf, 

Nicole O’Bleness Gray, Daniel Traetow, Sylvester Black, Elmahdi Elkhammas, 

Robert B. Kirkpatrick, Khalid Mumtaz 

1261 

Systolic Dysfunction Post-Liver Transplantation is associated 

with Increased All-Cause Mortality 

Michael Cheung 2 , She-Yan Wong 1 , Arun Mathew 3 , Dina Halegoua-De 

Marzio 1 ; 1 Medicine, Division of Gastroenterology and 

Hepatology, Thomas Jefferson University, Philadelphia, PA; 2 Medicine, 

Division of Cardiology, Lankenau Medical Center, Philadelphia, 

PA; 3 Medicine, Thomas Jefferson University Hospital, 


Introduction Left ventricular systolic dysfunction has been 

described after liver transplantation (LT), and is associated with 

a critically ill status, high pre-transplant Model of End-Stage 

Liver Disease (MELD) score and malnutrition. Previous studies of 

this condition have reported a high likelihood of ejection fraction 

recovery and survival at 1 year. The aim of this study was 

to evaluate all-cause mortality of systolic dysfunction post-LT at 

1 year and associated risk factors. Methods A retrospective 

chart review from a single liver transplant center was completed 

on all adult patients who received a liver transplant 

between 1/2002 and 1/2015. Patients with systolic dysfunction, 

defined as left ventricular ejection fraction (EF)


Disclosures: 

The following authors have nothing to disclose: Michael Cheung, She-Yan Wong, 

Arun Mathew, Dina Halegoua-De Marzio 

1262 

Downstaging or Bridging Therapy of Hepatocellular 

Cancer Does Not Increase Incidence of Biliary Complications 


Meera Ramanathan 2 , Mark Pedersen 2 , Myunghan Choi 3 , Anil B. 

Seetharam 1 ; 1 Banner Transplant and Advanced Liver Disease Center, 

University of Arizona College of Medicine-Phoenix, Phoenix, 

AZ; 2 Internal Medicine, University of Arizona College of Medicine-Phoenix, 

Phoenix, AZ; 3 Arizona State University College of 

Nursing and Health Innovation, Phoenix, AZ 

Background: Donor and vascular risk factors predisposing to 

bile duct ischemia are causes of biliary complications after liver 

transplant (LT). Manipulation of the hepatic artery prior to LT as 

often occurs in the management of hepatocellular cancer (HCC) 

may represent a novel risk factor for non anastomotic biliary 

complications. Aim: Evaluate effect of pre-LT hepatic artery 

based locoregional therapy on incidence of non anastomotic 

biliary complications after LT. Methods: Retrospective cohort 

study of consecutive LT recipients between 2009 and 2013. 

Incidence of non anastomotic biliary complications (stricture 

or leak) tabulated by review of radiologic or endoscopic cholangiogram. 

Univariate regression evaluated donor, recipient, 

surgical, and locoregional treatment variables associated with 

increased risk of biliary complication: cytomegalovirus (CMV) 

donor status, organ source (brain or cardiac death), donor 

age, native model for end stage liver disease (MELD) and body 

mass index (BMI) at LT, etiology of cirrhosis (viral vs. non-viral), 

warm ischemic time, cold ischemic time, need for take back surgery 

to control bleed in the 1 st month post LT, number of lesions 

at the time of HCC diagnosis, modality of arterial locoregional 

treatment (chemo vs radio-embolization) and number of treatment 

sessions. Variables of interest (p


1264 

Severity and Predictive Factors of Chronic Renal Dysfunction 


Yi Huang 1,2 , Leon A. Adams 1,2 , Oscar Arauz 2 , Peter W. Angus 3 , 

Marie Sinclair 3 , Graeme A. Macdonald 4 , Uthayanan Chelvaratnam 

4 , Alan J. Wigg 6 , Sze Pheh Yeap 6 , Nicholas A. Shackel 5 , 

Linda I. Lin 5 , Spiro C. Raftopoulos 2 , Geoff McCaughan 5 , Gary P. 

Jeffrey 2,1 ; 1 The University of Western Australia, Perth, WA, Australia; 

2 Sir Charles Gairdner Hospital, Perth, WA, Australia; 3 The 

Austin Hospital, Melbourne, VIC, Australia; 4 Princess Alexandria 

Hospital, Brisbane, Brisbane, QLD, Australia; 5 Royal Prince Alfred 

Hospital, Sydney, NSW, Australia; 6 Flinders Medical Centre, Adelaide, 

SA, Australia 

Background and aims: Renal dysfunction is one of the common 

complications after orthotropic liver transplantation (OLT). The 

aim of this study was to evaluate the prevalence and the predictive 

factors of chronic renal dysfunction and the requirement 

of dialysis post-OLT. Methods: Patients who underwent OLT 

in Australia in 2003-2009 were included. Exclusion criteria 

included: multi-organ transplantation, second OLT and a follow 

up time less than 6 months after OLT. Estimated glomerular 

filtration rate (eGFR), creatinine and dialysis status were 

recorded at the end of the follow up. Chronic renal dysfunction 

was defined as an eGFR less than 30 ml/min per 1.73 m 2 or 

the requirement of dialysis. Results: 474 patients were included 

in the final analysis: 338 (71%) were male and 136 (29%) 

were female, mean age was 50, one (0.2%) had dialysis pre- 

OLT, 71 (15%) had diabetes, 41 (9%) had hypertension. The 

mean follow up was six years (range: 0.5 to 10.3). 56 patients 

died during follow up: 23 patients died from liver related 

causes, 16 from non-HCC malignancy, 6 from infection, 2 from 

cardiovascular disease and 9 from other causes. Nine (2%) 

patients required a dialysis. Among patients without dialysis, 

the mean increase in creatinine post-OLT compared to pre-OLT 

was 22.8 umol/L. 114 (24%) patients had a creatinine reduction, 

308 (65%) patients had no change in creatinine level, 36 

(8%) patients had 1-fold creatinine increase, 7 (1.5%) patients 

had 2-fold increase and 8 (1.7%) patients had 3-fold increase. 

339 patients had eGFR measured at the end of follow up with 

a mean of 66 ml/min per 1.73 m 2 . 15 (4%) patients had an 

eGFR < 30 ml/min per 1.73 m 2 , 63(19%) patients had an 

eGFR < 50 ml/min per 1.73 m 2 and 261 (77%) patients had 

an eGFR ≥ 50 ml/min per 1.73 m 2 . 24 (7%) patients had 

chronic renal dysfunction. Univariate analysis found that diabetes, 

pre-OLT creatinine, hypertension, cholesterol were significantly 

correlated with chronic renal dysfunction with odds 

ratio of 3.3, 1.01, 3.5 and 0.7 respectively. Diabetes, pre-OLT 

creatinine, hypertension and days in ICU were significantly correlated 

with the requirement of dialysis post-OLT with odds ratio 

of 4.7, 1.01, 5.5 and 1.06 respectively. Multivariate analysis 

found that diabetes and pre-OLT creatinine were independent 

predictors for chronic renal dysfunction. Diabetes, pre-OLT creatinine 

and days in ICU were independent predictors for the 

requirement of dialysis post-OLT. Conclusion: This study showed 

a lower than expected rate of chronic renal dysfunction and 

dialysis post-OLT in Australia. Diabetes and pre-OLT creatinine 

were independent predictors for chronic renal dysfunction. 

Disclosures: 

Leon A. Adams - Patent Held/Filed: Quest diagnostics 



The following authors have nothing to disclose: Yi Huang, Oscar Arauz, Marie 

Sinclair, Graeme A. Macdonald, Uthayanan Chelvaratnam, Alan J. Wigg, 

Sze Pheh Yeap, Nicholas A. Shackel, Linda I. Lin, Spiro C. Raftopoulos, Geoff 

McCaughan, Gary P. Jeffrey 

1265 

Ohio Solid Organ Transplantation Consortium (OSOTC) 

criteria for liver transplantation in patients with alcoholic 

liver disease 

Kaveh Hajifathalian 1 , Zubin Arora 2 , Annette Humberson 2 , David 

S. Barnes 2 , Arthur J. McCullough 2 , Nizar N. Zein 2 , Bijan Eghtesad 

2 , Ibrahim A. Hanouneh 2 ; 1 Internal Medicine, Cleveland Clinic 

Foundation, Cleveland, OH; 2 Cleveland Clinic Foundation, Cleveland, 

OH 

Background A minimum of six to 12 months of abstinence and 

three months of participation in an alcohol rehabilitation program 

are typically required before patients with alcoholic liver 

disease are eligible for transplantation. Some patients are too 

ill to participate in a rehab program. The state of Ohio Solid 

Organ Transplantation Consortium (OSOTC) provides mechanisms 

for medically urgent patients to be listed for transplant 

after only one to three months of abstinence. Methods From 

our center’s liver transplant program, we selected patients with 

alcoholic liver disease who were deemed eligible for transplant 

based on OSOTC exception criteria. These are patients with 

low to medium risk of recidivism, who are willing to sign a contract 

to commit to a recovery program. Survival was compared 

between these patients and matched patients with alcohol liver 

disease from Organ Procurement and Transplant Network 

(OPTN) data records. Results 19 transplant patients with alcoholic 

liver disease who were listed based on OSOTC exception 

criteria were included in the analysis. Patients had a mean (SD) 

MELD score of 26 (8.8) before transplant. Median follow-up 

after transplant was 3.3 years (IQR 2.4-4.8). These patients 

were randomly matched based on exact MELD score to 38 

transplant patients from OPTN data records with a median 

follow up of 3.9 years months (IQR 1.6-6.5). At one year, 

cumulative survival rates (±SE) were 90±7% and 92±4% in 

OSOTC exception criteria and general liver transplant populations, 

respectively. The cumulative survival rates were 81±10% 

and 80±7% at three years. There was no difference in overall 

survival between two groups (p for log rank test=0.907). Only 

one patient from OSOTC group resumed drinking by the last 

follow-up visit. Conclusions Compared to traditional criteria 

for assessment of risk of recidivism in patient with alcohol liver 

disease, a careful selection process with more flexibility to evaluate 

eligibility on a case to case basis can lead to similar 

survival rates after transplant. 

Disclosures: 

The following authors have nothing to disclose: Kaveh Hajifathalian, Zubin 

Arora, Annette Humberson, David S. Barnes, Arthur J. McCullough, Nizar N. 

Zein, Bijan Eghtesad, Ibrahim A. Hanouneh 

1266 

Recipient Risk Factors for Acute Cellular Rejection After 

Orthotopic Liver Transplant in Patients with Average 

MELD More Than 25 

Mengyuan Liu 1 , Kelly Galen 1 , Grace Guzman 2 , Hoonbae Jeon 3 , 

Costica Aloman 1 ; 1 Division of Gastroenterology/Liver Diseases 

(MC716), University of Illinois at Chicago, Chicago, IL; 2 Pathology, 

University of Illinois at Chicago, Chicago, IL; 3 Transplant 

Surgery, University of Illinois at Chicago, Chicago, IL 

Background & Aims: The use of MELD score for liver organ 

allocation has resulted in transplanting sicker patients. Risk 

factors of acute cellular rejections (ACR) in liver transplant were 

reviewed in other previous studies before MELD era and in 

patients with MELD score less than 20. It is unclear whether 

the timing, severity, and risk factors of ACR have changed as 

a result of progressive increasingly MELD score at transplantation. 

Our aim was to assess ACR characteristics in our center,


where the average MELD score at transplantation is higher than 

previously published studies. Methods: This is a single center 

retrospective study designed to asses risk factors associated 

with ACR status post adult orthotopic liver transplant (OLT). 

Recipient demographics, preoperative clinical and laboratory 

data and cytomegalovirus (CMV) immune status were recorded 

for each transplant. Diagnosis of ACR was confirmed by biopsy 

using the Banff schema for grading liver allograft rejection. Univariate 

and multivariate regressions were preformed to look for 

variables that are significant predictors of ACR. Results: This 

study included 178 consecutive OLT patients transplanted in 

our center from January 2008 to June 2013 at the University 

of Illinois Hospital. Only 49.4% (88/178) of patients were 

Caucasians. In this study population, the average MELD at 

transplantation was 27.6, which is much higher than previously 

published studies that looked into risk factors of ACR. 

The average time from transplant to ACR diagnosis was 283.9 

days. Majority of ACR episodes were mild/moderate and only 

8.5% were graded as severe based on Banff classification. 

Of the twenty-two characteristics analyzed, creatinine was a 

significant predicator for ACR in both univariate and multivariate 

models. Patients with creatinine elevation greater than 2.7 

mg/dL have lower rates of ACR. Underlying liver etiology was 

also studied and patients with either primary biliary cirrhosis 

(PBC) or primary sclerosing cholangitis (PSC) have significantly 

higher rates of ACR. Conclusion: The study confirmed a change 

regarding the timing and severity of ACR in MELD era. Recipient 

characteristics such as elevated creatinine and PBC/PSC 

background may affect the risk of ACR and should be taken 

into consideration when managing immunosuppression. 

Disclosures: 

The following authors have nothing to disclose: Mengyuan Liu, Kelly Galen, 

Grace Guzman, Hoonbae Jeon, Costica Aloman 

1267 

Pre-Transplant Sarcopenia Does Not Reverse After Successful 


Trushar Patel, Valery Vilchez, Roberto Gedaly, Rashmi T. Nair, 

Anna Christina Dela Cruz, Terrence Barrett; University of Kentucky, 

Lexington, KY 

Objective: To study the evolution of pre-transplant Sarcopenia 

after successful Liver Transplantation and compare survival of 

sarcopenic and non-sarcopenic patients Methods: Patients who 

underwent liver transplantation between 2008 and 2013 at 

our center were analyzed. We used CT scan at third lumbar 

vertebrae to analyze with the SliceOmetic 5.0 software (Tomovision, 

Canada) which enables specific tissue demarcation using 

previously reported Hounsfield unit (HU) thresholds. Cutoffs for 

sarcopenia were based on a CT-based sarcopenia study for 

patients with malignancies (Skeletal Muscle Index (SMI)


significant CMV infection seen mostly within 1 st month in seropositive 

population. Post-Tx CMV infection does not correlate 

with pre-Tx CMV immunity. 

Disclosures: 

The following authors have nothing to disclose: Ekta Gupta, Viniyendra Pamecha, 

Nadeem Hasnain, Yogita Verma, Ajeet S. Bhadoria, Niteen Kumar, Shiv 

K. Sarin 

1269 

Good results of liver transplantation for decompensated 

cirrhosis with multi-organ failure in ICU: a retrospective 

study in two French centers 

Florent Artru 1 , Alexandre Louvet 1 , Isaac Ruiz 2 , Julien Labreuche 3 , 

Eric Levesque 2 , Philippe Ichai 2 , Sebastien Dharancy 1 , Guillaume 

Lassailly 1 , Emmanuel Boleslawski 4 , Gilles Lebuffe 5 , Audrey Coilly 2 , 

Eric Vibert 6 , Philippe Mathurin 1 , Didier Samuel 2 , Faouzi Saliba 2 ; 

1 Hepatology, CHRU Lille, Lille, France; 2 Hepatology, Hopital Paul 

Brousse, Villejuif, France; 3 Biostatistics Unit, CHRU Lille, Lille, 

France; 4 Surgery Transplant Unit, CHRU Lille, Lille, France; 5 Anaesthesiology 

Department, CHRU Lille, Lille, France; 6 Surgery Transplant 

Unit, Hopital Paul Brousse, Villejuif, France 

Resuscitation and transfer in intensive care unit (ICU) is often 

debated in patients with end-stage liver disease (ESLD), a condition 

associated with a very poor outcome. Few data are 

available on the results of liver transplantation (LT) in cirrhotic 

patients with multiorgan failure (MOF) and ESLD, admitted to 

the ICU. Aims: 1) to describe outcome of patients with ESLD 

and MOF transplanted while in ICU 2) to compare the outcome 

of these patients to those who were not hospitalized in 

ICU at time of LT and to those admitted to ICU but who did not 

benefit from LT. Methods: Patients were retrospectively selected 

from two French centers if they underwent LT for ESLD while in 

ICU with at least 2 non-hematological organ failures defined 

by SOFA score or if they had only liver failure but required 

mechanical ventilation at time of LT. We excluded combined 

liver-kidney transplantation. In order to evaluate the impact 

of MOF at time of LT, we performed a case-control study in 

which each patient with MOF was matched A) to 4 control 

patients transplanted outside ICU (matching on age and gender) 

B) to 1 (or 2 when possible) patients hospitalized in ICU 

with MOF, not transplanted (matching on age, gender, SOFA 

score). Results: From 2008 to 2014, 41 transplant recipients 

with a median age of 53.4 years (19.5% female) met the 

inclusion criteria. At time of LT, MELD score was 40 (95%CI: 

33-40), SAPS 2 score was 51 (49-63) and SOFA score was 

14 (13-15). Median duration of ICU stay before LT was 9 

days (5-12). Prior to LT, 39% had vasopressive therapy, 51% 

had mechanical ventilation and 51% had renal replacement 

therapy. Duration of stay after LT was 12 days in ICU and 43 

days in total. No PNF was observed. 6 patients underwent a 

surgical procedure during the first month. 32 patients (78%) 

developed bacterial infection post-LT, 12 (29%) CMV reactivation 

and 7(17%) fungal infection. The 1-year patient survival 

of the cohort was 85.04±5.6%. Based on the case-control 

study, this good survival at 1 year was not different from that 

of matched controls transplanted outside ICU (n=164): 85.04% 

vs. 87.04%, p=0.8. As expected, patients transplanted with 

MOF had a higher MELD score at time of LT as compared 

to patients transplanted outside ICU: 40 vs. 16 (p


MS was diagnosed at 3, 6 and 12 months post-LT respectively 

in 6/26 (23%), 7/23 (30%) e 7/16 (44%) patients. After 12 

months post-LT 11/16 patients (66%) had an HOMA2 test 

suggestive for IR, whereas at 6 months post-LT that finding 

was present in 10/23 (43%). No cardiovascular events were 

recorded in the study cohort. In conclusion, these data show 

that post-LT MS affects nearly half of LT recipients, starting early 

after LT. Lifestyle modifications, should be recommended to 

transplanted recipients, starting in the early post-LT period. This 

would facilitate prevention of body weight gain and the associated 

abnormalities, thus reducing incidence of post-LT MS and 

the related cardio-vascular events. 

Disclosures: 

The following authors have nothing to disclose: Veronica Pepe, Giacomo Germani, 

Alberto Ferrarese, Alberto Zanetto, Elena Nadal, Ilaria Bortoluzzi, Francesco 

P. Russo, Marco Senzolo, Umberto Cillo, Patrizia Burra 

Intraoperative Factors 

1271 

How intra-operative factors affect kidney recovery after 

simultaneous liver-kidney transplantation (SLK) 

Mario Spaggiari 1 , Mohamed Shaaban 2 , Galal El-Gazzaz 1 , Lisa 

Louwers 1 , Emmanouil Palaios 1 , Cristiano Quintini 1 , Federico N. 

Aucejo 1 , Koji Hashimoto 1 , Teresa Diago 1 , Masato Fujiki 1 , Bijan 

Eghtesad 1 , Charles M. Miller 1 , Mauricio Perilla 2 , Dympna Kelly 1 ; 

1 Cleveland Clinic, Cleveland, OH; 2 Department of Anesthesia, 

Cleveland Clinic Foundation, Cleveland, OH 

Introduction: The present study by analyzing our SLK population 

wants to identify the modifiable risk factors that affect 

kidney recovery after transplant with particular focus on the 

intraoperative events. Material and Methods: This a retrospective 

analysis performed between 2004 and 2014. The 

patient/graft survival of SLK were compared with the group of 

liver transplant alone (OLT) and kidney transplant alone (KT) 

in the same era. DGF was defined as the need of dialysis in 

the first week post-transplant. Results: 64 SLK were performed 

from 2004 to 2014.21.8% of patients (14/64) didn’t require 

dialysis at the time of the transplant. When compared to OLT 

(n=936) and KT (n=632) there was no significant differences 

in patient and graft survival. The incidence of DGF was higher 

in the SLK group (27%) compared to 20% in the KT (P= 0.3). 

Pre-operative factors associated with DGF were higher recipient 

BMI (p


1273 

Morbid Obesity and Diabetes (DM) are Associated with 

Increased Risk of Death on the Liver Transplant (LT) 

Waiting List 

Ani A. Kardashian 1 , Jennifer L. Dodge 2 , John P. Roberts 2 , Danielle 

Brandman 1 ; 1 Medicine, University of California, San Francisco, 

San Francisco, CA; 2 Surgery, University of California, San Francisco, 


Purpose: Obesity is a growing problem in LT candidates, paralleling 

the US obesity epidemic and the increase in LT for 

nonalcoholic steatohepatitis (NASH). While post-LT survival 

appears to be similar in obese and non-obese patients, data 

is scarce regarding risk of waitlist dropout in morbidly obese 

patients. We aimed to determine the impact of obesity on 

waitlist outcomes and evaluate predictors of dropout in those 

with morbid obesity or NASH. Methods: We retrospectively 

evaluated LT candidates listed between 3/02-12/13. We 

performed a competing risk analysis to evaluate predictors of 

removal or death on the waitlist. Variables with p-value40. Compared to those 

with BMI 25-29.9, patients with BMI≥40 were more likely to 

be female (46% vs 28%), diabetic (25% vs 18%), have NASH 

(35% vs 13%), and have shorter median waitlist times (161 vs 

208 days); all p


Disclosures: 

The following authors have nothing to disclose: Kazuhiro Takahashi, David 

A. Bruno, Marwan Kazimi, Atsushi Yoshida, Marwan S. Abouljoud, Gabriel 

Schnickel 

1275 

Post-Transplant Lymphoproliferative Disease is more 

common in primary sclerosing cholangitis patients after 

liver transplant 

Ahmed Abu Shanab 1,2 , Yasser Gayed 1 , Naeem Ullah 1 , Diarmaid 

D. Houlihan 1 , Aiden McCormick 1 ; 1 Liver Unit, St-Vincent University 

Hospital, Dublin, Ireland; 2 Internal Medicine, Menofiya University, 

Shebin Alkom, Egypt 

Background: Post transplant lymphoproliferative disorder (PTLD) 

is a well recognized complication of therapeutic immunosuppression 

in solid organs transplant recipients. It represents a 

broad spectrum of abnormalities ranging from a benign infectious 

mononucleosis like illness to malignant lymphoma and 

is associated with high mortality. Methods: A retrospective 

study was performed by collecting data of liver transplant (LT) 

patients in a single institution from December 1993 to December 

2014. Data was analyzed to identify PTLD patients and 

determine their demographic details, the indication for liver 

transplant, presenting symptoms, immunosuppression regimens 

and patient survival and PTLD outcome. Results: From a total of 

705 liver transplants recipients, 20 patients (2.8%) were diagnosed 

with PTLD. There were more males (n=17, 85%) than 

females (n=3, 15%), with median time from LT to diagnosis of 

PTLD was 83 months. The primary indication for LT of the PTLD 

cohort was PSC (n=8, 40%) of overall 67 PSC post LT patients. 

The rate of occurrence of PTLD in PSC was significantly higher 

than the overall LT patients with p value


The mean body mass index (BMI) at transplant was 27±4.2kg/ 

m 2 with mean albumin 2.96±0.8g/dL. The mean TPA was 

927.1±280mm 2 , which is much lower than the expected normal 

mean of 1211.9±242.3mm 2 . During the study time period, 

13 (54%) patients died at a mean of 1.2±2 years. Conclusion: 

Although LVSD is an uncommon complication post-LT, patients 

with high MELD and malnutrition maybe at highest risk. Sarcopenia, 

measured by TPA, maybe a better marker for malnutrition 

than traditional markers. BMI can be falsely elevated by 

volume overload, which can explain the normal BMI seen in 

our patient group. Sarcopenia should be studied further as a 

possible predictor for LVSD. Pre-operative nutritional optimization 

and close post-LT echocardiogram monitoring is advisable 

in this population at risk for increased mortality post-LT. 

Disclosures: 

The following authors have nothing to disclose: Arun Mathew, She-Yan Wong, 

Michael Cheung, Flavius Guglielmo, Cataldo Doria, David A. Sass, Dina Halegoua-De 

Marzio 

1277 

Risk Stratification for Optimal Timing of Liver Transplantation 

Following Repeated Locoregional Therapies in 

Patients With Hepatitis B-Related Hepatocellular Carcinoma 

Hwi Young Kim 1 , Won Kim 1 , Yong Jin Jung 1 , Hyeyoung Kim 2 , 

Nam-Joon Yi 2 , Kwang-Woong Lee 2 , Hae Won Lee 3 , Kyung-Suk 

Suh 2 ; 1 Department of Internal Medicine, Seoul National University 

Boramae Medical Center, Seoul, Korea (the Republic of); 2 Department 

of Surgery, Seoul National University Hospital, Seoul, Korea 

(the Republic of); 3 Department of Surgery, Seoul National University 

Boramae Medical Center, Seoul, Korea (the Republic of) 

BACKGROUND: Decision of timing for liver transplantation (LT) 

in patients with HCC often depends on the responses to pre-LT 

locoregional therapies especially in cases of organ shortage. 

However, prediction of favorable tumor biology before LT 

is difficult, and supporting data for decision of LT timing is 

scarce. The aims were to prognosticate HCC patients who 

underwent LT according to their responses to pre-LT locoregional 

therapies, and to shed light on the decision of optimal 

timing for LT. METHODS: Between January 2005 and 

December 2011, a total of 264 patients with hepatitis B-related 

HCC underwent LT. Excluding 52 patients with their 

tumors beyond Milan criteria, 212 patients were included. 

Responses to pre-LT locoregional therapies were assessed using 

the modified RECIST criteria. Pre-LT risk scores were generated 

as follows: [(No. of recurrence following radiofrequency 

ablation or ethanol injection + 2x(No. of progressive disease 

following transarterial chemoembolization(TACE)) + (No. of 

recurrence following previous response to TACE)]. Cox proportional 

hazard model was used to investigate prognostic 

factors for recurrence-free survival. RESULTS: Median age at 

the time of HCC diagnosis was 51 years, and male patients 

were 81.1%. Median time to LT was 6.5 months(interquartile 

range(IQR), 1.3-27.6). Median follow-up duration after LT 

was 55.3 months(IQR, 39.6-81.4). Median maximal diameter 

of the largest tumor was 2.0cm(range, 1.1-5). Hepatitis 

B viral DNA was 1875 IU/mL(median) at baseline. Baseline 

serum alpha-fetoprotein was 19 ng/mL(range, 1.8-23200). 

Baseline Child-Pugh classes were A in 101(47.6%) and B in 

61(28.8%), respectively. Before LT, 68 patients received at 

least one session of radiofrequency ablation or ethanol injection, 

and 100 patients received at least one session of TACE, 

respectively. Overall, 127 patients(59.9%) received pre-LT 

locoregional therapies. Pre-LT risk scores were as follows: 0, 

n=126(59.4%); 1-2, n=36(17.0%); 3-5, n=30(14.2%); ≥6, 

n=20(9.4%). From multivariable Cox analysis, pre-LT risk 

scores and maximal tumor diameter were independently significant 

prognostic factors for recurrence-free survival: risk score 

0vs.1-2, hazard ratio(HR)=1.422(95% confidence interval(CI), 

0.523-3.867), P=0.490); 0vs.3-5, HR=2.424(95%CI, 0.922- 

6.369; P=0.072); 0vs.≥6, HR=4.805(95%CI, 1.782-12.957; 

P=0.002); maximal diameter, HR=1.514(95%CI, 1.097- 

2.090; P=0.012). CONCLUSION: Increasing pre-LT risk score 

may represent a warning signal toward prompt LT rather than 

repeated locoregional therapies, especially in case of score≥6. 

Pre-LT risk score could be used as a surrogate marker of tumor 

biology for the decision of LT timing. 

Disclosures: 


GreenCross 

The following authors have nothing to disclose: Hwi Young Kim, Won Kim, Yong 

Jin Jung, Hyeyoung Kim, Nam-Joon Yi, Hae Won Lee, Kyung-Suk Suh 

1278 

Synergistic anticancer effect of metformin in combination 

with immunosuppressant on hepatocellular carcinoma 

cell lines 

Suk-Won Suh 1 , Kwang-Woong Lee 2 , Yoo-Shin Choi 1 ; 1 Department 

of Surgery, Chung-Ang University College of Medicine, Seoul, 

Korea (the Republic of); 2 Seoul National University College of 


After liver transplantation (LT), immunosuppression is needed 

to avoid rejection and graft loss, however, it can stimulate 

hepatocellular carcinoma (HCC) recurrence and progression. 

Previous studies have shown that metformin had an anticancer 

effect on several cancers, including HCC. The aim of this 

study was to evaluate the interactions between metformin and 

immunosuppressive agents including sirolimus, tacrolimus and 

mycophenolate mofetil (MMF) for antitumor activity. Three cell 

lines (Huh7, HepG2 and Hep3b) were tested. Cell viability 

was determined using a MTT assay and Western blot analysis 

for mammalian target of rapamycin (mTOR) pathway related 

proteins were performed to reveal their mechanism. Metformin 

and sirolimus had synergistic antiproliferative effect and sirolimus 

plus metformin supplemented with MMF also showed 

synergistic antiproliferative effect in specific HCC cells. Synergistic 

effect of metformin and sirolimus through the inhibition 

of mTOR and its down-stream, p70S6K, and p-4EBP1 were 

demonstrated. Metformin and sirolimus also showed synergistic 

effect for the down-regulation of Livin and Survivin expressions 

in HepG2 and Hep3b cells. In conclusion, metformin had synergistic 

interactions with sirolimus in terms of anticancer effects 

for HCC cells and the mechanism explaining this synergistic 

inhibition might be related with mTOR pathway. These results 

may provide a foundation for further studies for patients with 

HCC who underwent LT in clinical era. 

Disclosures: 


GreenCross 

The following authors have nothing to disclose: Suk-Won Suh, Yoo-Shin Choi


1279 

Evaluation of risk indices to determine 1 year graft survival 

after liver transplantation using the UNOS database 

Shahzad Ahmed, Vinay Sundaram; Cedars-Sinai Medical Center, 

Los Angeles, CA 

Aim: Given the shortage of viable organs available for liver 

transplantation (LT), relative to the supply, it is critical to ensure 

optimal post-transplant graft survival in recipients of LT. The 

Model for End-Stage Liver Disease (MELD) scoring systems was 

developed to determine 3-month survival prior to LT. Additional 

scoring systems have been developed to determine post-transplant 

outcomes including Donor Risk Index (DRI), Organ Patient 

Index (OPI) and Balance of Risk (BAR) score. The aim of this 

study was to evaluate the accuracy of these risk indices in evaluating 

post-transplant graft survival at 1 year. Methods: We 

analyzed the UNOS database from 2006-2009. We excluded 

patients under age 18 or who underwent multi-organ transplantation 

Results. A total of 24,388 patients were studied. Among 

these patients, 35% had hepatitis C virus, 15% had alcoholic 

liver disease and 12% had cholestatic liver disease (primary 

biliary cirrhosis and primary sclerosing cholangitis). Regarding 

1 year graft surivival, our findings indicated that all indices performed 

with modest accuracy as follows: MELD (>30/=16/


with adequate control of dyslipidemia in both groups. Blood 

glucose was higher in CNI (p


apeutic potential for the management of liver IRI in transplant 

recipients. 

Disclosures: 

Ronald W. Busuttil - Grant/Research Support: Bayer, Fujisawa, Novartis, Roche/ 

Genentech, NIH 

The following authors have nothing to disclose: Shi Yue, Changyong Li, Xingliang 

Zhou, Qilong Ying, Jerzy Kupiec-Weglinski, Bibo Ke 

1284 

The long noncoding RNA VL30-1 is a key regulator of 

liver injury via enhancement of HIF1a mediated inflammation 

Xinshou Ouyang, Sheng-Na Han, Irma Garcia-Martinez, Luke 

Cai, Richard A. Flavell, Wajahat Z. Mehal; Yale University, New 

Haven, CT 

Introduction: Inflammation is the key feature of liver injury in 

response to many insults. The inflammatory gene expression 

program is comprised of several specific transcription factors 

involving long noncoding RNAs (lncRNAs). HIF1a is well 

known regulator of inflammatory gene transcription, and is 

required for persistent liver inflammation. LncRNAs play important 

roles in diverse biological processes. However, the role of 

lncRNAs in regulating hepatic inflammation is not known. Aim: 

To identify lncRNA (s) that regulates liver inflammation and 

injury. Methods: Primary mouse macrophages were treated 

with bacterial lipopolysaccharide (LPS), and whole-transcriptome 

analysis (RNA-seq) was conducted. Numerous lncRNAs 

were differentially expressed including the short form of the 

endogenous retrotransposon VL30-1. Further analysis determined 

the functional role and molecular mechanisms of VL30-1 

in regulation of immune genes. This was done by siRNA knockdown,, 

transcriptome microarray, promoter luciferase assay, 

ELISA, RT-PCR, confocal imaging and RNA immunoprecipitation 

PCR (RIP)-PCR. The in vivo role of VL30-1 in liver injury 

was demonstrated by knockdown in the LPS/D galactosamine 

(LPS/D GlN) model. Results: VL30-1 is strongly induced by LPS 

in mouse macrophages (fold > 90 vs control). Stable depletion 

of VL30-1 in Raw264.7 cells by shRNA significantly down-regulated 

LPS induced Il1b mRNA and protein levels. IL-1b secretion 

was significantly reduced by VL30-1 depletion in mouse 

peritoneal macrophages in response to typical inflammasome 

activation (808+/-45 pg/ml in control vs 214+/-125 pg/ml 

in VL30-1 siRNA). VL30-1 depletion also markedly reduced 

mIL-1b luciferase activity under forced expression of HIF1a 

and HIF2a. Conversely, forced VL30-1 expression dose-dependently 

trans-activated mIL-1b reporter luciferase activity, 

confirming that VL30-1 augments HIF1a mediated IL-1b 

transcription. RIP-PCR assay showed a direct VL30-1 binding 

with HIF1a. VL30-1 co-localized with HIF1a by confocal 

imaging analysis. Depletion of VL30-1 in mice significantly 

decreased LPS/ D-GalN induced serum IL-1b protein concentration 

(188+/-76 pg/ml in control vs 23+/- 26 pg/ml) as 

well as reduced liver damage as judged by morphology and 

serum ALT level (418+/-297 in control vs 151+/- 76 in VL30-1 

siRNA). Conclusions: These studies reveal a critical role of the 

lncRNA VL30-1 as a broad-acting regulatory component in 

the control of HIF1a induced inflammatory pathway activation 

in mouse macrophages and liver injury. This occurs via direct 

interaction of VL30-1 and HIF-1a, and up-regulation of HIF-1a 

mediated inflammatory transcripts. 

Disclosures: 

The following authors have nothing to disclose: Xinshou Ouyang, Sheng-Na Han, 

Irma Garcia-Martinez, Luke Cai, Richard A. Flavell, Wajahat Z. Mehal 

1285 

Galectin-3 mediates NLRP3 inflammasome signaling 

and contributes to pathogenesis of primary biliary cirrhosis 

Jijing Tian 1,2 , Guo-Xiang Yang 1 , Fu-Tong Liu 1 , M. Eric Gershwin 

1 , Natalie J. Torok 1 , Joy Jiang 1 ; 1 UCDMC, Sacramento, CA; 

2 Research Center for Eco-environmental Sciences, Chinese Academy 

of Sciences, Beijing, China, Beijing, China 

Macrophages play a significant role in early inflammatory 

changes in primary biliary cirrhosis (PBC). However, their 

role in modulating fibrogenic responses has not been evaluated. 

We and others have shown that macrophages are 

a major source of galectin-3, a profibrogenic lectin. As the 

pathogenesis of PBC is not well understood, we hypothesized 

that galectin-3 is required to activate inflammasome signaling 

contributing to inflammation, fibrosis and progression in PBC. 

Methods: Liver tissues from PBC patients and healthy controls; 

and from the dnTGFβRII transgenic mouse model of PBC and 

wt controls were collected for RT-PCR and Western blot to analyze 

the expression of galectin-3, NLRP3, ASC and IL-1β. The 

cleavage of caspase-1 and IL-1β in PBC patients was examined 

by Western blots. Primary hepatic macrophages were 

isolated from wt and the galectin3 -/- mice, and treated with 

Deoxycholic Acid (DCA) with/without recombinant galectin-3. 

RT-PCR was done to analyze the activation of inflammasome-related 

transcripts; immunofluorescence (IF) to detect 

the activation of inflammasomes; and immunoprecipitation (IP) 

to detect the association of galectin-3 and NLRP3. DnTGFβRII/ 

gal3 -/- mice were generated by crossing the dnTGFβRII mice 

with galectin3 -/- mice and the liver tissues were analyzed for 

inflammasome activation and fibrosis. Results: The transcripts 

of galectin-3, NLRP3, ASC and IL-1β significantly increased in 

the livers of PBC patients (N=4, p


1286 

Kupffer cells signature in alcoholic steatohepatitis in 

mice encompasses loss of signalling/ chaperone and 

enrichment of cytoskeleton proteins in the lipid rafts 

Angela Dolganiuc 1 , Tracie Lo 2 ; 1 Medicine/GI, University of Florida, 

Gainesville, FL; 2 New York Medical College, New York, NY 

The mechanisms of alcoholic liver disease (ALD) are complex 

and largely related to dysfunctional Kupffer cells (KCs). Here 

we set to define the role of lipid rafts (LR) in alcohol exposed 

KCs. Mice were fed alcohol-containing or pair-fed Lieber-De- 

Carli Diet for 4 weeks. Plasma and liver tissue were analyzed 

for Tg and ALT by enzymatic methods; RNA by PCR. KCs were 

isolated by tissue digestion, gradient centrifugation and adherence 

to plastic. KCs and their model cell line RAW 264.7 

were further stimulated in vitro with LPS; cell migration was 

assayed with RTCA DP method; detergent-resistant membranes 

fractions, representative of lipid rafts (LR), were separated by 

gradient centrifugation and analyzed with LC-MS/MS and 

MALDI-TOF mass spectrometry. Alcohol consumption, unlike 

pair-fed diet, was associated with elevated serum ALT and liver 

Tg, suggestive of ALD steatohepatitis. KCs of alcohol-fed mice 

produced higher TNFa, suggestive of imbalanced signaling, 

and migrated slower on plastic, suggestive rigidity of cytoskeleton, 

compared to alcohol naive counterparts. The analysis 

of Log2 relative expression of proteins in the lipid raft protein 

profile resulted from mass spectrometry revealed that 93% of 

all detected proteins were similar in resting alcohol naïve and 

alcohol-treated RAW 264.7 cells. Among proteins enriched 

in the LRs of alcohol-treated cells compared to alcohol naïve 

counterparts were cytoskeleton-related proteins which are 

known to anchor to the cytoplasmic side of the LRs, including 

tubulin, beta actin, moesin, talins and synaptic protein VAT- 

1, alongside with those exposed on the extracellular side of 

the LRs including Fermitin family homolog pleckstrin homology 

domain-containing family C member 1. In contrast, we 

observed a significant loss of a cohort of signaling proteins in 

LPS-activated alcohol-treated RAW 264.7 cells compared to 

alcohol naïve counterparts; among most prominently depleted 

were iron-sulfur binding proteins, PARK chaperones, nontransforming 

monomeric GTP-binding proteins and sorting nexins. 

Only few signaling proteins were upregulated in alcohol 

exposed LPS-activated cells however these did not reach statistical 

significance. The RNA levels and the total cellular content of 

all proteins above were comparable in KCs of alcohol exposed 

mice and their pair-fed control counterparts. In conclusion, our 

novel results suggest impaired recruitment of signaling proteins 

and chaperones likely leading to functional impairment of lipid 

rafts in activated alcohol-exposed KCs, possibly secondary to 

increased LR rigidity secondary to enrichment of cytoskeleton 

proteins. These results point to novel therapeutic targets in ALD. 

Disclosures: 

The following authors have nothing to disclose: Angela Dolganiuc, Tracie Lo 

1287 

IL28B genetic variants determine the extent of monocyte-induced 

activation of NK cells in hepatitis C 

Benjamin Krämer 1 , Beatriz Sastre Turrión 2 , Claudia Finnemann 1 , 

Philipp Lutz 1 , Andreas Glässner 1 , Franziska Wolter 1 , Pavlos 

Kokordelis 1 , Dominik J. Kaczmarek 1 , Felix Goeser 1 , Christian P. 

Strassburg 1 , Ulrich Spengler 1 , Jacob Nattermann 1 ; 1 Department of 

Internal medicine I, Univeritiy of Bonn, Bonn, Germany; 2 Infectious 

Diseases Department, Hospital Ramón y Cajal - IRYCIS, Madrid, 

Spain 

Background: Genetic variants in IL28B (IFNλ3) in combination 

with the natural killer (NK) cell-associated killer cell Ig-like 

receptors (KIR) gene KIR2DS3 synergistically increased the 

risk of chronic infection over either factor alone. A proposed 

direct functional link between NK cells and λ-IFNs could not 

be confirmed since in line with the known limited expression of 

IFN-λR1, the natural λ-IFN receptor, on epthelial cells. NK cells 

do not express IFN-λR1 and do not respond to λ-IFNs. Thus, the 

synergism between IL28B genotype and NK cell activity must 

be mediated by indirect mechanisms. Here, we studied differential 

interactions between monocytes and NK cells which can 

contribute to the observed synergism between polymorphisms 

in the KIR and IFNλ genes. Methods: A total of 74 HCV(+) 

patients and 80 healthy controls were enrolled into this study. 

IL28B (rs 12979860) genotypes were determined by rtPCR. 

Peripheral NK cells and monocytes, respectively, were isolated 

using MACS technology. Total PBMC, isolated monocytes, and 

NK cells were stimulated with IL28B, the TLR7/8 agonist R848, 

or a combination of both. NK cell IFN-g response was analysed 

by FACS. IL12 and IL-18 secretion of monocytes was studied by 

ELISA. In blocking experiments anti-IL12 and/or anti-IL18 were 

used. Results: Following stimulation of total PBMCs with R848 

we found CD56Bright NK cell IFN-g responses to be associated 

with the IL28B genotype, with carriers of a T/T genotype 

displaying the lowest frequency of IFN-g(+) CD56 Bright NK 

cells (C/C vs. T/T, p


1288 

High baseline expression of type 1 interferon (IFN)-inducible 

genes involved in cell-autonomous immunity 

predicts mortality in patients with decompensated alcoholic 

cirrhosis 

Marc Pineton de Chambrun 1 , Emmanuel Weiss 1 , Pierre-Emmanuel 

Rautou 2 , Magali Fasseu 1 , Mikhael Giabicani 1 , Rakhi Maiwall 

3 , Dominique Valla 2,1 , Didier Lebrec 2,1 , Francois Durand 2,1 , 

Pierre de la Grange 4 , Sophie Lotersztajn 1,2 , Richard Moreau 1,2 ; 

1 UMR S1149, Center for Research on Inflammation (CRI), Inserm 

and Paris Diderot University, Clichy, France; 2 DHU Unity, Service 

d’Hépatologie, Hôpital Beaujon, Clichy, France; 3 Hepatology 

Department, Institute of Liver and Biliary Science, New Delhi, 

India; 4 Genosplice, Paris, France 

Background & Aims: Although systemic inflammation is 

believed to be a major driver of mortality in patients with 

decompensated alcoholic cirrhosis, its mechanisms are unclear. 

We hypothesized that baseline expression levels of genes 

involved in cell-autonomous immunity (most of which are type 

1 IFN-inducible) and/or of genes encoding secreted inflammatory 

cytokines or chemokines by circulating mononuclear cells 

in patients with cirrhosis may be related to patient outcome. 

Thus, we measured the baseline gene expression in peripheral 

blood mononuclear cells (PBMCs) from patients with decompensated 

alcoholic cirrhosis and investigated their relationship 

with the risk of death. Methods: PBMCs were isolated in 98 

non infected patients (MELD score: 19 (15-24)) and 50 healthy 

subjects. Using RT-qPCR, we monitored 52 type 1 IFN-inducible 

genes and 10 IFN-non-inducible genes encoding ‘traditional’ 

secreted inflammatory cytokines (e.g., IL6) or chemokines (e.g., 

CXCL1). We also used a prespecified IFN score (a composite 

of type 1 IFN-inducible genes: OAS2, MX2, DDX58, GBP4, 

IFIH1, TRIM22, IFIT1, CXCL10). Gene expression in ‘cirrhotic 

cells’ was normalized using expression in ‘healthy’ cells. 

Patients were then followed-up for 10±12 months, until death, 

transplantation or the end of the study. Results: In univariate 

analysis, a higher MELD score, higher baseline expression of 

9 IFN-inducible genes as well as higher IFN score values were 

significant predictors of death. In contrast, cytokine or chemokine 

gene expressions were not significantly related to death. In 

bivariate analysis including the IFN and MELD scores, only the 

former significantly predicted death (RR=3.58; 95% CI, 1.18- 

10.91; P=0.02). These results were mainly due to the elevated 

intrinsic prognostic value of OAS2 (RR=2.49; P=0.04) and 

MX2 (RR=1.35; P=0.01). It is interesting to note that IFI35 and 

IFI44, two genes not included in the IFN score were significant 

predictors of death independent of the MELD score. Additional 

experiments performed in blood from patients with early septic 

shock with (n=7) or without (n= 7) cirrhosis showed that the 

IFN score was 2.3 higher in patients with than in those without 

cirrhosis (due to higher values of OAS2, IFIT1, GBP4). Conclusions: 

In PBMCs from patients with decompensated alcoholic 

cirrhosis, higher baseline levels of type 1 IFN-inducible genes 

involved in cell-autonomous immunity, but not expression of 

genes encoding secreted inflammatory cytokines/chemokines, 

are highly predictive of the risk of death. Deregulation of type 1 

IFN-inducible gene expression in circulating immune cells may 

play a role in the mechanisms resulting in death from cirrhosis. 

Disclosures: 


Dominique Valla - Advisory Committees or Review Panels: Sequana medical; 

Consulting: IRIS 



Pierre de la Grange - Management Position: GenoSplice 

The following authors have nothing to disclose: Marc Pineton de Chambrun, 

Emmanuel Weiss, Magali Fasseu, Mikhael Giabicani, Rakhi Maiwall, Didier 

Lebrec, Sophie Lotersztajn, Richard Moreau 

1289 

Proportion of Intrahepatic CD56 Bright Natural Killer Cells 

Correlates with Well-preserved Liver Function 

Erin H. Doyle 1 , Adeeb Rahman 2 , Arielle L. Klepper 1 , Sang Kim 3 , 

Brandy M. Haydel 4 , Sander S. Florman 5 , M. Isabel Fiel 6 , Thomas 

D. Schiano 5 , Andrea D. Branch 1 ; 1 Department of Liver Diseases, 

Icahn School of Medicine at Mount Sinai, New York, NY; 2 Human 

Immune Monitoring Core, Icahn School of Medicine at Mount 

Sinai, New York, NY; 3 Department of Anesthesiology, The Mount 

Sinai Hospital, New York, NY; 4 Center for Translational Transplant 

Research, Icahn School of Medicine at Mount Sinai, New York, 

NY; 5 Recanati Miller Transplantation Institute, The Mount Sinai 

Hospital, New York, NY; 6 Department of Pathology, The Mount 

Sinai Hospital, New York, NY 

Background/Aim: HCV-related liver damage is largely 

immune-mediated, but immunosuppression increases liver damage. 

We investigated this paradox, seeking innate immune 

cells whose prevalence correlates with well-preserved liver 

function. Methods: Immune cells were isolated from 5-25 g 

of tissue from 20 HCV-infected liver transplant patients using 

mechanical disruption and enzymatic digestion. Peripheral 

blood (obtained prior to surgery) and liver mononuclear 

leukocytes were isolated on ficoll gradients and analyzed 

by FACS and microarray. 8 subsets of innate immune cells 

were enumerated. Pearson’s correlation coefficient was used 

to find subsets whose prevalence correlated with liver status. 

Microarrays were performed on RNA from cells with or without 

exposure to TLR ligands. Results: Intrahepatic CD56 Bright / 

CD16 - NK cells were strongly correlated with well-preserved 

liver function and inversely related to markers of liver dysfunction: 

MELD score (R 2 =0.41, p=0.007), total bilirubin (R 2 =0.74, 

p


These findings have direct implications for our understanding 

of the roles of IL-22 and IL-22BP in regulating liver immunity. 

Disclosures: 

The following authors have nothing to disclose: Wagdi Almishri, Julie Deans, 

Mark Swain 

Disclosures: 




The following authors have nothing to disclose: Erin H. Doyle, Adeeb Rahman, 

Arielle L. Klepper, Sang Kim, Brandy M. Haydel, Sander S. Florman, M. Isabel 

Fiel 

1290 

Activation of hepatic innate immunity markedly alters 

the IL-22/IL-22R1/IL-22BP axis within the liver: Implications 

for hepatic immunity. 

Wagdi Almishri, Julie Deans, Mark Swain; Medicine, University of 

Calgary, Calgary, AB, Canada 

IL-22 is an innate cytokine that has been broadly implicated 

in the regulation of hepatic immunity, showning both pro- and 

anti-inflammatory effects in animal models of liver injury. IL-22 

exerts its’ biological functions by interacting with its’ receptor 

IL-22R1 which is widely expressed on epithelial cells and 

hepatocytes. Importantly, IL-22 activity is regulated by an 

endogenous inhibitor, IL-22BP, which binds IL-22 and prevents 

its’ biological function. Therefore, we performed a series of 

experiments to delineate alterations that occur in the IL-22/ 

IL-22BP/IL-22R1 axis in the liver, using a mouse model of 

hepatic innate immune activation due to NKT cell activation. 

In addition, we determined changes in IL-22BP and IL-22R1 

expression in liver biopsy samples obtained from patients with 

hepatitis B and C, PBC, PSC and autoimmune hepatitis (AIH). 

Results: We found (by flow cytometry) the rapid and striking 

recruitment of IL-22 producing innate lymphoid cells (ILC3’s) 

into the liver, followed later by other IL-22 producing immune 

cells, including Th22 cells. In addition, using IL-22 knockout 

mice we demonstrate a proinflammatory role of IL-22 in the 

liver during innate immune activation. The recruitment of IL-22 + 

immune cells was paralleled by recruitment of IL-22BP + immune 

cells into the liver, and the marked up-regulation of IL-22BP 

expression in hepatocytes. Moreover, despite our findings of 

a robust influx of IL-22 + immune cells into the liver after NKT 

cell activation, bioavailable hepatic IL-22 levels actually significantly 

decreased in the 1-2 days after activation of hepatic 

innate immunity. Within the liver IL-22BP expression in recruited 

immune cells and in hepatocytes was largely dependent upon 

activation of the inflammasome. Viral and autoimmune liver diseases 

in patients have been linked to the activation of NKT cells 

and an enhancement of hepatic innate immunity. Therefore, we 

examined IL-22BP and IL-22R1 expression in liver biopsy specimens 

obtained from patients with liver disease, and found a 

striking increase in the hepatic expression of both IL-22BP and 

IL-22R1 in both viral and autoimmune liver diseases (ie. PBC, 

PSC, AIH, hepatitis B and C). Conclusion: These observations 

highlight a tight co-regulation of the IL-22/IL-22BP/IL-22R1 

axis during hepatic innate immune responses, and suggest 

that IL-22BP plays an important role in regulating IL-22 bioactivity 

within the liver in the context of innate immune activation. 

1291 

Impaired TRAF6 methylation and TLR responses in liver 

tissue and circulating monocytes from patients with 

spontaneous bacterial peritonitis 

Irina Tikhanovich, Jody C. Olson, Ryan Taylor, Brian Bridges, Kenneth 

Dorko, Benjamin R. Roberts, Steven A. Weinman; University 

of Kansas Medical Center, Kanas City, KS 

Patients with cirrhosis have a number of abnormalities in their 

response to infection including failure to clear bacteria from 

the peritoneal space or blood and an exaggerated systemic 

inflammatory response. Cirrhotic patients with a history of 

spontaneous bacterial peritonitis (SBP) have particularly severe 

defect in bacterial clearance. Our previous work has shown 

that the arginine methyltransferase PRMT1 and the demethylation 

enzyme JMJD6 regulate innate immune signaling 

pathways by controlling the arginine methylation of TRAF6, 

a critical ubiquitin ligase for TLR responses. We found that a 

decrease in the PRMT1/JMJD6 ratio results in a loss of TRAF6 

methylation causing preactivation of TLR pathways but reduced 

responses to bacterial antigens. The AIM of this study was to 

determine whether cell type specific defects in TRAF6 arginine 

methylation occur in cirrhosis and can contribute to susceptibility 

to infections. METHODS: Liver tissue sections and peripheral 

blood monocytes from patients without cirrhosis and cirrhotic 

patients with or without a history of SBP were analyzed for 

the levels of PRMT1, TRAF6 methylation and cytokine production 

in response to LPS using IHC staining, western blotting 

and proximity ligation assay. RESULTS: PRMT1 to JMJD6 protein 

ratios was lower in total liver protein from cirrhosis than 

normal donors (2.2±1.1 vs 0.68±0.43) and lower in patients 

with history of SBP compared to those with no history of SBP 

(0.92±0.48 vs 0.43±0.16). Tissue staining revealed that in 

patients with SBP compared to cirrhotics without an SBP history, 

PRMT1 was lower in non-parenchymal cells (p-value 0.029), 

and JMJD6 was higher both in hepatocytes and non-parenchymal 

cells (p-values 0.0009 and 0.025 respectively). PRMT1/ 

JMJD6 ratio was significantly lower both in hepatocytes and 

non-parenchymal cells of patients with history of SBP (p-values 

0.044 and 0.005 respectively). TRAF6 methylation levels 

were directly analyzed by PLA and were similarly lower in 

livers of patients with history of SBP. Similar to the situation 

in liver, blood monocytes from patients with a history of SBP 

showed a similar defect in PRMT1 levels compared to monocytes 

from patients with cirrhosis without SBP (p-value 0.032) 

and the associated decrease in PRMT1/JMJD6 ratio correlated 

with decreased TRAF6 methylation and cytokine production 

in response to LPS. CONCLUSION: PRMT1 and JMJD6 levels 

control TRAF6 methylation and antibacterial innate immune 

responses. TRAF6 methylation is decreased in multiple cell 

types of patients with a history of SBP, which may contribute to 

the abnormal infection responses seen in those patients. 

Disclosures: 

Jody C. Olson - Advisory Committees or Review Panels: Baxter 


The following authors have nothing to disclose: Irina Tikhanovich, Ryan Taylor, 

Brian Bridges, Kenneth Dorko, Benjamin R. Roberts


1292 

Implants of matrix-embedded endothelial cells rescue 

ischemic tissue and liver engraftment in hepatectomized 

mice 

Pedro Melgar-Lesmes 1 , Mercedes Balcells 2,1 , Elazer R. Edelman 1,3 ; 

1 Edelman Lab, Institute for Medical Engineering and Science, 

Massachusetts Institute of Technology, Cambridge, MA, USA, 

Cambridge,, MA; 2 Bioengineering department, Institut Químic de 

Sarrià, Ramon Llull Univ, Barcelona, Spain; 3 Cardiovascular Division, 

Brigham and Women’s Hospital, Harvard Medical School, 

Boston, MA 

Background and aims: Ischemia during liver transplantation 

promotes a cascade of cellular injury responses that lead to 

inflammation, cell death, and organ dysfunction. Matrix-embedded 

endothelial cells (MEECs) have immunomodulatory 

effects in vitro and in vivo. Therefore we investigated the benefits 

of implants of MEECs as modulators of inflammation and 

regeneration after liver engraftment in mice. Methods: Partial 

hepatectomy (70%) was performed by excising the left lobe 

and half the median lobe of the mouse liver. Four groups of 

10 animals were distributed as follows: 1) acellular matrices 

in the interface between the remaining median lobe and an 

autograft from the left lobe; 2) MEECs between the remaining 

median lobe and an autograft from the left lobe; 3) acellular 

matrices between the remaining median lobe and an allograft 

from the left lobe of another mouse; 4) MEECs between the 

remaining median lobe and an allograft from the left lobe of 

another mouse. Vascular architecture was analysed using angiography 

with FITC dextran 2000 kDa by fluorescent multiphoton 

microscopy 7 days post-op. Hepatic DNA fragmentation 

and apoptosis were quantified in the median lobe and grafts 

by TUNEL assay and Western Blot of activated caspase 3, 

respectively. Serum markers of liver damage Alanine Aminotransferase 

(ALT) and Aspartate Aminotransferase (AST) were 

quantified in all animal groups. The phenotype of lymphocyte 

subsets in the liver after implantation of allografts was quantified 

by gene expression of Th1 (interferon gamma: INFγ; interleukin 

2: IL-2) and Th2 (interleukin 4: IL-4; interleukin 10: IL-10) 

genes by Real-Time PCR. Results: Implants with MEECs created 

a functional vascular splice between the ischemic median lobe 

and autografts or allografts improving the rate of liver donor 

regeneration by 15% compared to acellular controls. MEECs 

prevented apoptosis by 85% in donor liver lobes, by 85% 

in autologous grafts and by 79% in allogeneic engraftments. 

As a result, serum levels of ALT and AST were significantly 

reduced after autologous or allogeneic engraftments. The beneficial 

immunomodulatory effects of MEECs promoted allograft 

immunotolerance expressed as a significant reduction of Th1 

(INFγ and IL-2) and increase of Th2 (IL-4 and IL-10) cytokine 

expression. Conclusions: Implants of MEECs are endothelial 

cell-based scaffolds with potential to be used to improve current 

surgical procedures in liver transplantation and to reduce 

ischemic and inflammatory disorders. Their application may 

contribute obtaining functional liver grafts of adequate size for 

the recipient without subjecting the donor to undue risk. 

Disclosures: 

The following authors have nothing to disclose: Pedro Melgar-Lesmes, Mercedes 

Balcells, Elazer R. Edelman 

1293 

Identification of liver monocytic myeloid-derived suppressor 

cells and elucidation of their roles in non-alcoholic 

fatty liver disease 

Liying Yao, Masanori Abe, Teruki Miyake, Yoshiko Nakamura, 

Yusuke Imai, Yohei Koizumi, Takao Watanabe, Osamu Yoshida, 

Masashi Hirooka, Yoshio Tokumoto, Bunzo Matsuura, Yoichi 

Hiasa; Department of Gastroenterology and Metabology, Ehime 

Universiy Graduate School of Medicine, Ehime, Japan 

Background/Aim: Myeloid-derived suppressor cells (MDSCs) 

comprise a heterogeneous population of myeloid cells and are 

recognized as suppressors of T cell functions. In mice, these 

cells identified by the co-expression of CD11b and Gr-1 surface 

markers. Previous studies revealed that liver MDSC exhibit 

phenotypic and functional diversity; however, information 

regarding their characteristics and mechanisms of suppression, 

reported in these studies, was conflicting in nature. Immune 

responses may contribute to the pathogenesis of non-alcoholic 

fatty liver disease (NAFLD); however, little is known regarding 

the role of myeloid regulatory cells. In this study, we characterized 

the phenotype of liver MDSCs in a murine model of 

NAFLD and explored the mechanism underlying their immunosuppression. 

Methods: C57BL/6 mice were fed a normal 

diet (ND) or a high-fat diet (HFD) for 12 months. Different subtypes 

of CD11b + Gr-1 + cells were sorted from liver non-parenchymal 

cells by FACS. CD11b + Gr-1 + cells were co-cultured 

with T cells in the presence of anti-CD3 beads or allogeneic 

dendritic cells, and suppressive functions were investigated 

using a carboxyfluorescein succinimidyl ester assay. Nitric 

oxide (NO) concentrations in culture supernatants were measured 

using Griess reagent. In some experiments, L-NIL, an 

inducible NO synthase (iNOS) inhibitor, was added at the 

start of the culture. Results: In the livers of HFD-fed mice, the 

frequency of CD11b + Gr1 dim cells (monocytic myeloid cells), 

but not CD11b + Gr1 hi cells, was higher than that in the livers 

of ND-fed mice over time. CD11b + Gr1 dim cells were divided 

into SSC high and SSC low populations. The frequency of SSC low- 

CD11b + Gr1 dim cells increased in the livers of HFD-fed mice 

to a greater extent than that observed in the livers of ND-fed 

mice. In addition, SSC low CD11b + Gr1 dim cells suppressed T cell 

proliferation in a dose-dependent manner; however, this suppression 

was not observed in SSC low CD11b + Gr1 high cells. We 

also found that SSC low CD11b + Gr1 dim cells expressed iNOS 

after co-culture with T cells, and the supernatants obtained from 

the co-culture of SSC low CD11b + Gr1 dim cells with T cells had 

higher concentrations of NO. By adding iNOS inhibitor L-NIL 

to the SSC low CD11b + Gr1 dim cells and T cells co-culture, T cell 

proliferation was restored. Conclusion: SSC low CD11b + Gr1 dim 

cells represent authentic monocytic MDSCs in the liver, and 

their numbers were increased in the livers of NAFLD mice. The 

suppressive function of monocytic MDSCs was dependent on 

NO production by iNOS. These cells might have a role in the 

pathogenesis of NAFLD. 

Disclosures: 

The following authors have nothing to disclose: Liying Yao, Masanori Abe, Teruki 

Miyake, Yoshiko Nakamura, Yusuke Imai, Yohei Koizumi, Takao Watanabe, 

Osamu Yoshida, Masashi Hirooka, Yoshio Tokumoto, Bunzo Matsuura, Yoichi 

Hiasa


1294 

Chronic alcohol consumption skews hepatic dendritic 

cell homeostasis and reveals a novel potential cellular 

source of TNFα 

Holger Fey, Costica Aloman; Division of Gastroenterology/Liver 

Diseases (MC716), University of Illinois at Chicago, Chicago, IL 

Background/Aim: Alcohol abuse is one of the leading causes 

of chronic liver disease and liver-related mortality. Tumor necrosis 

factor alpha (TNFα) is considered a central mediator in the 

pathogenesis of alcoholic liver damage. Dendritic cells (DC) 

are mononuclear phagocytes and critical regulators of innate 

and adaptive immunity. Several populations of DC coexist in 

mouse and humans: classic DC (cDC) and plasmacytoid DC 

(pDC). In this study we investigated the effects of chronic alcohol 

exposure on hepatic DC homeostasis and their contribution 

to the hepatic cytokine environment. Methods: Alcohol (EtOH) 

was delivered in the drinking water to C57BL/6 mice at 20% 

v/v (Meadows-Cook diet). Hepatic DC sets and bone marrow 

(BM) progenitors of the myeloid lineage were analyzed by flow 

cytometry. Circulating growth factors involved in DC development 

(Flt3L, GM-CSF, M-CSF) were measured by ELISA. TNFα 

production was assessed by intracellular cytokine staining of 

hepatic leukocytes isolated after in vivo toll-like receptor (TLR) 

stimulation with TLR4 and TLR9 agonists. Results: C57BL/6 

mice receiving alcohol in drinking water for 12 weeks have a 

70% increase (P


recruitment under conditions of shear stress identified inflammatory 

cells that migrated into HSEC and subsequently exhibited 

novel cell-to-cell crawling. This process was dependent on 

IFN-γ and occurred predominantly in HSEC when compared 

to vascular endothelium. Expression of chemokine transcripts 

and transcellular permeability were similar in both endothelia, 

whereas distinct differences were identified in junctional molecule 

expression, with HSEC lacking occludin and expressing 

lower levels of JAM-A than vascular endothelium. In support 

of these findings immunofluorescent analysis of human liver 

tissue demonstrated CD3+/CD4+ cells within sinusoidal endothelial 

cells. Conclusion - We demonstrate a novel behaviour 

of lymphocyte migration which occurs during interactions with 

primary human HSEC. This process appears to be mediated by 

the unique junctional expression pattern of HSEC and microenvironmental 

stimuli such as IFN-γ. We believe this is a novel 

step in the adhesion cascade that could have implications for 

treating chronic inflammatory liver disease and modulating 

intrahepatic immunity. 

Disclosures: 

David H. Adams - Advisory Committees or Review Panels: GSK, Proximagen; 

Grant/Research Support: Takeda, Biotie Therapies, Novimmune, ChemoCentryx, 

Novimmune, Biotie Therapies; Patent Held/Filed: biotie therapies, Biotie Therapies, 

Biotie Therapies, Biotie Therapies, Biotie Therapies, Biotie Therapies, Biotie 

Therapies, Biotie Therapies 

The following authors have nothing to disclose: Shishir Shetty, Daniel A. Patten, 

Chris J. Weston 

1297 

Inhibition of the FGL2:FcγRIIB Immunosuppressive Pathway 

Restores Antiviral T and B Cell Responses During 

Chronic Viral Infection 

Ramzi Khattar, Olga Luft, Kaveh Farrokhi, Vanessa Rojas Luengas, 

Hassan Sadozai, Gary Levy, Nazia Selzner; University Health 

Network, Toronto, ON, Canada 

Background and Aims: HBV and HCV utilize host immunosuppressive 

pathways to limit antiviral T cell responses and promote 

the generation of dysfunctional T cells in a process termed 

T cell exhaustion. Recent reports have implicated the upregulation 

of immunosuppressive cytokines and involvement of Tregs 

as causative agents of persistent HBV and HCV infection. FGL2 

is a potent immunosuppressive effector molecule of CD4 + C- 

D25 + TIGIT + FOXP3 + Treg. The role for FGL2 in establishing T 

cell exhaustion in chronic viral infection caused by LCMV Cl-13 

was assessed. Methods: Fgl2 +/+ and fgl2 -/- mice were infected 

with 2x10 6 PFU of LCMV Cl-13. FGL2 was measured in plasma 

by ELISA. Frequencies of LCMV specific T cells were determined 

following stimulation with viral peptide. Neutralizing 

antibody titers were evaluated by plaque reduction assay. Viral 

titers in plasma and liver were assessed by an LCMV focus 

forming assay. Fgl2 +/+ were treated with neutralizing antibodies 

toward FGL2 and FCγRIIB and subsequently infected with 

2x10 6 PFU of LCMV Cl-13. LCMV antiviral immune responses 

were measured. Results: Following infection with 2x10 6 PFU of 

LCMV Cl-13, plasma levels of FGL2 in Fgl2 +/+ mice increased 

(51.6±2.4 ng/mL) reaching maximum levels on day 7 pi 

(135.6±7.0 ng/mL; P


αEβ7 suggest homing to the peri-biliary region where LI MAIT 

cells are activated in response to bacteria-exposed BEC in an 

MR1 dependent manner, leading to liver injury through their 

release of cytolytic enzymes and inflammatory cytokines. 

Disclosures: 






The following authors have nothing to disclose: Hannah C. Jeffery, Bonnie Van 

Wilgenburg, Kathryn Stirling, Krishan Parekh, Sheree Roberts, Ayako Kurioka, 

Stuart Hunter, Paul Klenerman, Ye H. Oo 

1299 

Alterations in liver dendritic cell subtypes during non-alcoholic 


Eva-Carina Heier, Hannes Borchardt, Henrike Julich, Christoph 

Eckert, Niklas Klein, Thomas Tschernig, Veronika Lukacs-Kornek; 

Department of Medicine II, Saarland University, Homburg, Germany 

BACKGROUND/AIMS: Dendritic cells (DCs) represent a heterogeneous 

subpopulation that is primarily identified as 

CD11c + MHCII + cells in the liver. However, these markers are 

not exclusively identifying DCs rather they are relevant to a 

mixture of myeloid/lymphoid cells. Nevertheless, CD11c + cells 

limited inflammation by reducing the destructive effect of innate 

cells and promoting the clearance of cellular debris in non-alcoholic 

steatohepatitis (NASH). The various DC subsets have different 

roles in liver homeostasis and divergent factors determine 

their functional operation during inflammation (Lukacs-Kornek 

V. J Hepatol. 2013 Nov;59(5):1124-6). In order to clarify the 

nature and subset of DCs involved in NASH, the study aimed 

to characterize the CD11c + myeloid infiltrates and dissect the 

role of the various DC subtypes during disease progression. 

METHODS: CD11c + cell infiltrate was analyzed in methionine 

choline deficient (MCD) diet induced liver steatohepatitis in the 

progression and regression phase. Using multicolor FACS analyses, 

the myeloid compartment was mapped during disease 

progression and the individual DC subtypes, and precursor 

DCs were identified and distinguished from monocytes, macrophage 

and neutrophils. RESULTS: During disease progression, 

CD11c + cells were increased and among these cells, the 

CD11c + CD11b + myeloid DC subtype increased an average of 

2.1 fold in 1wk and 2.2 fold in 5wks MCD treated animals. 

Not only the myeloid but also the lymphoid DC subtypes followed 

similar changes. Accordingly, the frequency of lymphoid 

DCs (CD103 + CD11c + CD11b - ) increased during disease progression 

(2.2 and 2.3 fold increase in 1wk and 5wks MCD 

treated animals). During regression of steatohepatitis, these 

changes were reduced and after 1wk of regression period 

DC parameters were similar to control animals. Functionally, 

both DC subtypes produced high amount of inflammatory cytokines 

(e.g. TNFα), (30%-40% of myeloid DCs and 10% of lymphoid 

DCs). Besides, the precursor DC population was more 

abundant in 1wk MCD treatment and further elevated in 5wks 

MCD treatment. CONCLUSIONS: During MCD diet induced 

steatohepatitis both DC subtypes show increased abundance 

and exhibited similar fold changes suggesting the importance 

of both DC subtypes in the pathomechanism of NASH. DC 

changes are paralleled by alterations observed in the precursor 

DC population suggesting the relevance of local DC differentiation 

during steatohepatitis. Studies were supported by 

the Alexander von Humboldt Foundation – Sofja Kovalevskaja 

Award 2012 to VLK. 

Disclosures: 

The following authors have nothing to disclose: Eva-Carina Heier, Hannes 

Borchardt, Henrike Julich, Christoph Eckert, Niklas Klein, Thomas Tschernig, 

Veronika Lukacs-Kornek 

1300 

Soluble CD163 plasma concentration is related with 

necroinflammatory activity and liver function impairment 

in compensated cirrhotic patients and is not 

related with bacterial translocation 

Francesco Figorilli 2,1 , Karen Louise Thomsen 2 , Jane Mac- 

Naughtan 2 , I.Jane Cox 3 , Alba Moratalla 4 , Isidora Ranchal 2 , 

Holger J. Møller 5 , Rajeshwar Mookerjee 2 , Nathan Davies 2 , Rajiv 

Jalan 2 ; 1 Universita’ di Cagliari, Cagliari, Italy; 2 Liver Failure 

Group, UC, Institute for Liver and Digestive Health, UCL Medical 

School, Royal Free Campus, London, United Kingdom; 3 nstitute of 

Hepatology, London, Foundation for Liver Research, 69-75 Chenies 

Mews, London, United Kingdom; 4 Liver unit, Hospital General 

Universitario de Alicante, CIBER de Enfermedades Hepáticas y 

Digestivas (CIBERehd), Alicante, Spain; 5 Department of Clinical 

Biochemistry, Aarhus University Hospital, Aarhus, Denmark 

Background: CD163 is a scavenger receptor expressed on 

monocytes and macrophages e.g. Kupffer cells in the liver. 

Plasma levels of soluble CD163 (sCD163, normal range 0.89- 

3.95 mg/L) are associated with macrophage activation and 

grade of portal hypertension and provide prognostic information 

in cirrhosis. The mechanism underlying this increase 

in sCD163 is not clear. In this study we aimed to determine 

whether there was a relationship between the level of sCD163 

and gut permeability and markers of bacterial translocation 

or severity of liver disease. Methods: 66-clinically-stable liver 

cirrhosis out-patients were included (Male 69.7%; Age 57 

±9.15; Alcoholic liver disease 57.6%). Exclusion criteria were 

a Child Pugh score >10 and clinical or microbiological evidence 

of infection 7 days prior to enrolment. sCD163 plasma 

level was measured by an ELISA test, gut permeability using 

the lactulose/rhamnose test, bacterial DNA by PCR reaction 

and LPS concentration was obtained by HEK-Blue LPS Detection 

Kit 2s. Urine was collected for proton magnetic resonance 

spectroscopy ( 1 HMRS). Patients were divided accordingly to 

their sCD163 level (3.95). For the statistical analysis 

we used Chi-square, Student T-test, Man Whitney test and 

Spearman correlation. Results: The median value of sCD163 

was 4.33mg/L with a range of 1.44-15.43. We did not find 

any significant correlation between the levels of sCD163, gut 

permeability (0.028, range 0.002-0.112), or LPS concentration 

(0.02ng/ml, range 0-14.1). There was no difference in 

the prevalence of bacterial DNA positivity in patients with an 

abnormal value of sCD163 (11.1% vs. 2.6%). 39/66 (59.1%) 

patients with a sCD163>3.95mg/L showed a significant 

higher MELD value (9.0 vs. 7.5) and higher levels of AST (43 

vs. 30 U/L), ALP (102 vs. 70U/L), GGT (98 vs. 48 U/L) and 

lower level of platelets (107 vs. 178 x 10 9 ). Child-Pugh B was 

more common in patients with high sCD163 levels (17,9 vs. 

0%)). No significant differences were observed between the 

two groups regarding aetiology, alcohol consumption, drug 

use, comorbidity, previous episode of liver decompensation or 

infection in the previous month. At 1 HMRS analysis, patients 

with sCD163>3.95 showed no significant difference in urinary 

level of hippuric acid, formate, citrate or lactate. Conclusion: 

Our results suggest that Kupffer cell activation is an early 

event in the pathogenesis of chronic liver disease, which is 

not dependent of gut permeability or bacterial translocation. 

sCD163 concentration might reflect the grade of necroinflammation 

and liver function impairment in compensated cirrhotic 

patients.


Disclosures: 

Holger J. Møller - Grant/Research Support: Danish Council for Strategic 

Research; Independent Contractor: IQ-Products, NL; Patent Held/Filed: Aarhus 

University; Stock Shareholder: Affinicon Aps 

Rajiv Jalan - Consulting: Ocera Therapeutics, Conatus; Grant/Research Support: 

Grifols, Gambro; Patent Held/Filed: Yaqrit, Cyberliver 

The following authors have nothing to disclose: Francesco Figorilli, Karen Louise 

Thomsen, Jane MacNaughtan, I.Jane Cox, Alba Moratalla, Isidora Ranchal, 

Rajeshwar Mookerjee, Nathan Davies 

1301 

Hepatitis B X-interacting protein (HBXIP) negatively 

regulates IFN-β production and antiviral response by 

promoting proteasomal degradation of TANK-binding 

kinase 1 

Hang Zhang; Basic medical, Hangzhou Normal University, Hangzhou, 

China 

TANK-binding kinase 1 (TBK1) plays an essential role in Tolllike 

receptor (TLR)– and retinoic acid–inducible gene I (RIG-I)– 

mediated induction of type I interferon (IFN; IFN-α/β) and host 

antiviral responses. How TBK1 activity is negatively regulated 

remains largely unknown. We report that Hepatitis B virus X-interacting 

protein (HBXIP) promotes proteasomal degradation of 

TBK1 and inhibits RIG-I-induced IFN-β signaling. HBXIP knockdown 

resulted in augmented activation of IFN regulatory factor 

3 (IRF3) and enhanced expression of IFN-β in RIG-I-activated 

primary hepatocyte cells, whereas overexpression of HBXIP 

had opposite effects. Consistently, HBXIP impaired vesicular 

stomatitis virus (VSV) infection–induced IRF3 activation and 

IFN-β production and promoted VSV replication. HBXIP negatively 

regulated the cellular levels of TBK1 by directly binding 

to and promoting K48-linked polyubiquitination of TBK1. Therefore, 

we identified HBXIP as a negative regulator in RIG-I–triggered 

antiviral responses and suggested HBXIP as a potential 

target for the intervention of diseases with uncontrolled IFN-β 

production. 

Disclosures: 

The following authors have nothing to disclose: Hang Zhang 

1302 

Toll-like receptor 9 and B-cell activating factor signals 

induce impaired immunological memory in cirrhosis 

Hiroyoshi Doi 1 , David E. Kaplan 3 , Eiichi Hayashi 1 , Jun Arai 1 , Risa 

Omori 1 , Manabu Uchikoshi 1 , Kenichi Morikawa 4 , Junichi Eguchi 2 , 

Takayoshi Ito 2 , Hitoshi Yoshida 1 ; 1 Gastroenterology, Showa University, 

Shinagawa, Japan; 2 Digestive Disease Center, Showa University 

Koto Toyosu Hospital, Toyosu, Japan; 3 Gastroenterology, 

University of Pennsylvania, Philadelphia, PA; 4 Gastroenterology 

and Hepatology, Hokkaido University, Sapporo, Japan 

[Background] Cirrhosis is an immunocompromised state and 

their immune cells are thought to be impaired. We previously 

reported peripheral CD27+ memory B-cells in cirrhotic patients 

are decreased and dysfunctional compared to those in even 

non-cirrhotic chronic liver disease patients. We also found Tolllike 

receptor (TLR) 4 and TLR 9 signals play important roles for 

B-cell activation and survival. (Doi et al. Hepatology 2012). 

Clinically, we confirmed elevated immunoglobulin levels in cirrhosis 

regardless of the etiology. The abnormal humoral immunity 

could be caused by antigen non-specific way such as TLR 

signals and results in polyclonal B-cell activation. The purpose 

of this study is to investigate the impact of antigen non-specific 

signals on immunological change and immunodeficiency 

in cirrhosis. [Methods] We isolated peripheral blood mononuclear 

cells (PBMC) and serum from whole blood. Then we 

measured memory B-cell and memory T-cell defined by CCR7 

and CD45RA expressions using PBMC by flow cytometry. As 

we previously reported, B-cell stimulants such as TLR ligands 

increased in cirrhotic plasma probably due to bacterial translocation 

and we investigated TLR 4 and TLR 9 expressions on 

B-cell. B-cell activating factors (BAFF and APRIL) in sera were 

measured by ELISA. To analyze B-cell signaling, we negatively 

isolated CD19+ B-cell from healthy donor PBMC and cultured 

the B-cell for 72 hours with LPS (TLR4 agonist), CpG-ODN 

(TLR9 agonist), anti-IgG/A/M (B-cell receptor agonist) and 

combinations. Then we measured activation marker expressions 

and apoptosis by flow cytometry. [Results] In accordance 

with our previous study, cirrhotic patients (CIR) had decreased 

peripheral memory B-cell compared to non-cirrhotic chronic 

liver disease patients (CLD). (CLD vs CIR; p=0.021) They also 

showed lower frequency of both CD4+ and CD8+ effector 

memory T-cells. (p= 0.003, 0.027, respectively) TLR9 but not 

TLR4 expression on memory B-cell was increased in cirrhotic 

patients. (p=0.043) We also found serum BAFF but not APRIL 

level was elevated as well. (p=0.003) Memory B-cell frequency 

tended to be associated with the TLR 9 expression and BAFF 

levels. (p=0.016, 0.025, respectively) These findings are more 

apparent in advanced cirrhosis. In vitro, B-cell stimulants shown 

above didn’t make much impact on CD27 expression but 

CD27- B-cells, which relatively dominant in cirrhosis, tended 

to be apoptotic by TLR 4 and TLR 9 stimulations. [Conclusion] 

Cirrhotic patients especially in advanced stage have elevated 

BAFF and TLR9 expression on memory B-cell. Those antigen 

non-specific signals have an impact on dysfunctional immunological 

memory observed in cirrhosis. 

Disclosures: 

David E. Kaplan - Grant/Research Support: Bayer Pharmaceuticals, Inovio Pharmaceuticals 

The following authors have nothing to disclose: Hiroyoshi Doi, Eiichi Hayashi, 

Jun Arai, Risa Omori, Manabu Uchikoshi, Kenichi Morikawa, Junichi Eguchi, 

Takayoshi Ito, Hitoshi Yoshida 

1303 

Low Dose Zinc Sulfate (220mg) Supplementation for 

Three Months Normalizes Zinc Levels, Endotoxeima, 

Pro-Inflammatory/Fibrotic Biomarkers & Improves Clinical 

Parameters in Alcoholic Cirrhosis– A Double-Blind 

Placebo Controlled – (ZAC) Clinical Trial 

Mohammad K. Mohammad 1,2 , Keith C. Falkner 1 , Ming Song 1 , 

Craig J. McClain 1,2 , Matthew C. Cave 1,2 ; 1 Hepatology, gastroenterology 

and nutrition, university of louisville, Louisville, KY; 2 Hepatology, 

Robely Rex Veteran Hospital, Louisville, KY 

Purpose: Zinc deficiency occurs in human subjects with alcoholic 

cirrhosis (AC), and zinc supplementation attenuates liver 

injury/inflammation in murine models of alcoholic liver disease. 

The aim of this NIH-funded clinical trial was to determine 

if zinc sulfate therapy improves serologic biomarkers of liver 

injury/inflammation & clinical status in AC Methods: 22 consented 

subjects with Child-Pugh class A-B alcoholic cirrhosis 

were randomized to placebo (n=11) or zinc sulfate 220 mg 

daily (n=11) in the single center, IRB approved, double-blind, 

placebo-controlled clinical trial. 10 non-drinking, healthy volunteers 

were recruited as controls (HC). Serum cytokines (IL-1β, 

IL-2, IL-4, IL-6, IL-8, IL-10, IL-17, IL-18, IFN-γ, PAI-1, MCP-1 & 

TNF-α) and whole blood ex-vivo unstimulated and lipopolysacharide-stimulated 

(LPS) cytokine production were measured 

by Luminex, while TGF-β & LPS were measured by ELISA. Differences 

between means were evaluated by t-test, p < 0.05 

was considered significant Results: Of the 22 AC subjects, 

20 subjects completed 3 months treatment: placebo (n=10);


zinc (n=10). Demographic variables were similar between 

groups. At baseline, means for Zinc, AST, ALT,Albumin, CTP 

& MELD were 56.9±14; 42.3±15.88; 17.5±10.11; 3.5±0.7; 

6.9±1.22; 9.4±2.11 in the Zinc group, and 62.58±22.52; 

51.7±24.42; 27.3±17; 3.9±0.4, 5.6±0.9; 8.5±0.85 for 

placebo. There were more subjects with active EtOH drinking, 

varices & ascites (7/9/4) in the zinc arm vs. (2/6/1) 

in the placebo At baseline, when compared to HC, AC had 

increased mean serum LPS, TNF-α, IL-6, IL-8 while decreased 

mean Zinc. AC patients had increased mean ex-vivo unstimulated 

& LPS-stimulated production (Priming) of IL-6, IL-8, IL-18 

and TNF-α. No differences were observed for IL-1β, IL-2, IL-4, 

IL-10, IL-17 and IFN-γ in AC vs. HC with & without LPS stimulation. 

At 3 months, mean Zinc, LPS, TGF-β, TNF-α, IL-18 & Child- 

Pugh significantly improved in the Zinc group, while only IL-18 

improved in the placebo arm. In the zinc group, ex-vivo unstimulated 

& LPS stimulated production of TNFα & IL-6 normalized 

at 3 months together with unstimulated IL-18 & LPS stimulated 

production of IL-1β, no difference was observed in placebo. 

There were no side effects for zinc treatment Conclusions: Subjects 

with alcoholic cirrhosis had increased biomarkers of liver 

injury, and inflammation compared to healthy controls which 

improved with zinc supplementation. To our knowledge, this 

was first human study to demonstrate the safety & efficacy of 3 

months Zinc supplementation in improving zinc levels, endotoxemia, 

pro-inflammatory biomarkers (TNF-α, TGF-β, IL-1β, IL-6 

and IL-18) & clinical status in alcoholic cirrhosis 

Disclosures: 








The following authors have nothing to disclose: Mohammad K. Mohammad, 

Keith C. Falkner, Ming Song 

1304 

Acrolein, a lipid-derived aldehyde, is a critical pathogenic 

mediator and potential therapeutic target for alcoholic 

liver disease 

Wei-Yang Chen 1 , Jingwen Zhang 1 , Shirish Barve 1 , Craig J. 

McClain 1,2 , Swati Joshi-Barve 1 ; 1 Department of Medicine/GI, University 

of Louisville, Louisville, KY; 2 Robley Rex VAMC, Louisville, 

KY 

Purpose: Alcoholic liver disease (ALD) remains a major cause 

of morbidity and mortality, with no FDA approved therapy. 

Chronic alcohol consumption causes a pro-oxidant environment 

in the liver and increases hepatic lipid peroxidation. Acrolein 

is the most reactive and toxic aldehyde generated by lipid 

peroxidation, and is known to form covalent protein adducts. 

We have shown that acrolein exposure in hepatocytes triggers 

endoplasmic reticulum (ER) stress, which is an important etiologic 

factor in ALD. This study (i) investigates the pathogenic 

role of acrolein in alcohol-induced hepatic ER stress, steatosis, 

and liver injury in experimental ALD, and (ii) tests acrolein 

elimination/scavenging (using hydralazine) as a potential 

therapeutic strategy against ALD. Methods: We used in vitro 

(rat hepatic H4IIEC cells) and in vivo (chronic+binge (NIAAA) 

alcohol feeding in C57Bl/6 mice) models to study the role of 

acrolein in ALD. Also, the potential protective effects of the 

acrolein scavenger, hydralazine, were examined in vitro and 

in vivo. Alcohol-induced acrolein accumulation was detected 

by immunostaining for acrolein-protein adducts. The effects of 

alcohol-induced acrolein were assessed on (i) hepatic steatosis 

(H&E and Oil Red O); (ii) ER stress (ATF3, ATF4, GRP78, 

GRP94 mRNA and protein); (iii) pro-apoptotic signaling (activation 

of JNK and caspase-12 and pro-apoptotic CHOP); (iv) 

cell death (MTT) and hepatocyte apoptosis (TUNEL); (v) liver 

injury (serum ALT and AST). Results: We demonstrate that alcohol 

exposure resulted in substantial hepatic accumulation of 

acrolein-protein adducts, which was dependent on alcohol 

metabolism via cytochrome P4502E1 and alcohol dehydrogenase. 

Alcohol-induced acrolein accumulation led to hepatic 

ER stress and upregulation of ATF3 and ATF4, with minimal 

induction of the protective chaperones GRP78 and GRP94. 

A concomitant increase was seen in proapoptotic signaling 

with activation of JNK and caspase12, and CHOP, resulting 

in significant hepatic steatosis, hepatocyte cell death and liver 

injury. Acrolein was able to mimic the in vivo adverse effects 

of alcohol in cultured H4IIEC cells. Importantly, hydralazine, a 

known acrolein scavenger, protected against alcohol-induced 

ER stress, steatosis, apoptosis and liver injury, both in vitro and 

in vivo. Conclusions: Our study demonstrates that acrolein is a 

major mediator of alcohol-induced hepatic ER stress, cell death 

and liver injury. Notably, our study also shows that removal/ 

clearance of acrolein by scavengers may have therapeutic 

potential in attenuating the adverse effects of alcohol consumption, 

and preventing ALD. 

Disclosures: 





The following authors have nothing to disclose: Wei-Yang Chen, Jingwen Zhang, 

Swati Joshi-Barve 

1305 

Intestinal HIF-1α Deletion Exacerbates Alcohol-induced 

Hepatic Steatosis Mediated by Intestinal Dysbiosis and 

Barrier Dysfunction 

Tuo Shao 1 , Liming Liu 1 , Fengyuan Li 1 , Lihua Zhang 1 , Craig J. 

McClain 1,2 , Wenke Feng 1 ; 1 Department of Medicine/GI, University 

of Louisville, Louisville, KY; 2 Robley Rex VAMC, Louisville, KY 

Background: Alcoholic liver disease (ALD) is characterized by 

increased gut permeability and bacterial translocation. Hypoxia 

Induced Factor 1α (HIF-1α) has been implicated in transcriptional 

regulation of intestinal barrier integrity and inflammation. 

However, the link between intestinal HIF-1α regulation and ALD 

is unclear. We tested our hypothesis that HIF-1α plays a critical 

role in gut microbiota homeostasis and the maintenance of 

intestinal barrier integrity in a mouse model of ALD. Methods: 

Wide type (WT) and intestinal-specific HIF-1α knockout mice 

(HIF-1α -/- ) were pair-fed modified Lieber-DeCarli liquid diets 

containing 5% (w/v) alcohol (AF) or isocaloric maltose dextrin 

(PF) for 24 days. Blood samples were collected for determination 

of ALT and AST, endotoxin level and bacteria DNA. 

The liver and intestine tissues were used for histology staining, 

gene expression and hepatic triglyceride (TG) content measurements. 

Ex vivo ileal permeability was determined. Intestinal 

microbiota were determined in cecel samples Results: Alcohol 

feeding significantly increased serum levels of ALT, AST and 

LPS in the WT mice, indicating liver injury and increased endotoxemia 

by alcohol exposure. These elevations were more pronounced 

in HIF1α -/- mice. Histology examination revealed that 

hepatic lipid accumulation was increased by alcohol exposure, 

and it was exaggerated in HIF1α -/- mice. Liver triglyceride 

concentrations were increased by alcohol exposure in the WT 

mice and further elevated in the KO mice, and these were associated 

with up-regulation of hepatic SREBP-1c and down-regulation 

of PPAR-α. Intestinal HIF-1α deletion resulted in a marked


increase in hepatic IL-6 and IL-1β expression and neutrophil 

infiltration as showed by CAE staining, which were associated 

with TLR4 up-regulation. In the HIF-1α -/- mice, alcohol exposure 

disrupted intestinal structure integrity as shown by H&E staining 

of ileal sections. 16s RNA analysis of the cecal and fecal 

samples from HIF1α -/- mice exposed to alcohol showed an elevated 

dysbiosis. PCR analysis revealed that alcohol exposure 

decreased the expression of tight junction gene, occludin, and 

CRAMP, P-gp, and ITF, which are HIF-1α transcription target 

genes and important for gut microbiota homeostasis and intestinal 

barrier integrity. Ex vivo assay demonstrated that HIF1α 

-/- 

increased alcohol-induced ileal permeability. Conclusions: 

Our results demonstrated that intestinal HIF-1α is required for 

the adaptation response to alcohol exposure-induced changes 

in intestinal microbiota and barrier function leading to elevated 

endotoxemia and hepatic steatosis and injury. 

Disclosures: 




The following authors have nothing to disclose: Tuo Shao, Liming Liu, Fengyuan 

Li, Lihua Zhang, Wenke Feng 

1306 

Fibroblast growth factor 21 is critically involved in alcohol-induced 

hepatic lipid metabolism and adipose tissue 

lipolysis in mice 

Cuiqing Zhao 1 , Liming Liu 1 , Ying Yang 1 , Craig J. McClain 1,2 , 

Wenke Feng 1 ; 1 Department of Medicine/GI, University of Louisville, 

Louisville, KY; 2 Robley Rex VAMC, Louisville, KY 

Purpose: Fibroblast growth factor 21 (FGF21) is a novel 

metabolic regulator produced primarily in the liver that regulates 

hepatic fat metabolism in various models of metabolic 

syndromes. Alcoholic liver disease (ALD) is characterized by 

hepatic lipid accumulation. Here, we sought to determine 

the role of FGF21 in mice exposed to alcohol and to discern 

underlying mechanisms. Methods: Male FGF21 knockout (KO) 

and control (WT) mice were fed either the Lieber DeCarli diet 

containing 5% alcohol (AF) or an isocaloric diet (PF). For the 

chronic alcohol exposure, mice were fed for 4 weeks. For the 

chronic-binge alcohol exposure, mice were fed for 12 days, 

followed by a single dose of alcohol (5g/kg) or isocaloric 

maltose dextrin treatment by gavage. Mice were sacrificed 

6 hours later. One group of AF mice were administered with 

recombinant human FGF21 (rhFGF21) for the last 5 days. 

Liver steatosis and inflammation and epididymal white adipose 

tissue (eWAT) lipolysis were investigated. Results: Alcohol 

exposure induced plasma FGF21 elevation. FGF21 knockout 

exacerbated chronic alcohol-induced hepatic steatosis and 

liver injury, which was associated with increased activation 

of genes involved in lipogenesis mediated by SREBP1c and 

decreased expression of genes involved in fatty acid (FA) 

β-oxidation mediated by PGC1α. Chronic alcohol exposure 

induced a moderate adipose tissue lipolysis in eWAT. rhFGF21 

administration reduced alcohol-induced hepatic steatosis and 

inflammation in WT mice. Surprisingly, in the chronic-binge 

model, alcohol consumption-induced fatty liver is blunted in 

the KO mice. Chronic-binge alcohol exposure-induced in situ 

hepatic lipogenesis and FA β-oxidation were comparable to 

that in chronic alcohol-exposed mice. However, FGF21 deficiency 

markedly reduced chronic-binge alcohol exposure-induced 

eWAT lipolysis, which may contribute to hepatic FA 

uptake. Further analysis showed that in the KO mice alcohol 

induced significant elevation in systemic catecholamine, which 

is known to promote adipose lipolysis. rhFGF21 administration 

increased alcohol- induced fat loss in parallel with an increase 

of circulating norepinephrine concentration in KO mice. Conclusion: 

Our results demonstrated that the role of FGF21 in ALD 

is alcohol exposure model dependent. Lack of FGF21 exacerbates 

chronic alcohol exposure-induced fatty liver, and this 

exaggeration is overridden in chronic-binge alcohol exposed 

mice through a reduction of adipose lipolysis mediated by 

systemic catecholamine release involving sympathetic nervous 

system activation. Pharmacologic strategies targeting hepatic 

FGF21 elevation and reduction in adipose tissue lipolysis are 

warranted. 

Disclosures: 




The following authors have nothing to disclose: Cuiqing Zhao, Liming Liu, Ying 

Yang, Wenke Feng 

1307 

Early prediction of response in patients with severe 

acute alcoholic hepatitis by using Lille model on day 4 

Mauricio Garcia-Saenz de Sicilia 1 , Chitharanjan Duvoor 2 , Jose 

Altamirano 4,5 , Veronica Prado 3,4 , Juan Caballeria 3,4 , Andres 

Duarte-Rojo 1 ; 1 Gastroenterology and Hepatology, University of 

Arkansas for Medical Sciences, Little Rock, AR; 2 Internal Medicine, 

University of Arkansas for Medical Sciences, Little Rock, AR; 3 Liver 

Unit, Hospital Clinic, Barcelona, Spain; 4 Institut d’Investigacions 

Biomediques August Pi I Sunyer (IDIBAPS), Barcelona, Spain; 5 Vall 

d’Hebron Institut de Recerca, Barcelona, Spain 

BACKGROUND: Corticosteroids are the mainstay of treatment 

for severe alcoholic hepatitis (SAH) associated with a lower 

28-day mortality. Lille model (LM) can identify non-responders 

after 7 days of treatment, however this requires longer 

exposure and increases the risk of infection. We aimed to 

evaluate LM at day 4(LM4) of treatment with corticosteroids 

in comparison to day 7(LM7). METHODS: We performed a 

retrospective analysis of a multinational cohort with SAH based 

on a discriminant function (DF)>32. Response to corticosteroids 

was assessed with LM7 and LM4 according to the validated 

cutoff value (CUV>0.45). ROC-curves were constructed to 

determine an ideal CUV for LM4, and to compare accuracy 

of LM4, LM7, MELD, and ABIC. Logistic regression models 

were constructed to predict 90-day mortality. RESULTS: A total 

of 132 (83.9%) out of 162 patients with SAH received corticosteroids. 

Mean age was 47.87±10.58, 98 (60.49%) were 

males, and BMI was 27.58±6 kg/m 2 . On admission the mean 

bilirubin was 17±10 mg/dL, creatinine 0.97 (0.7-1.8) mg/ 

dL, INR 1.92±0.63. Severity parameters on admission were 

DF 58.34±33.38, MELD 24.98±8.79, and ABIC 8.18±1.56. 

Although mean LM4 and LM7 were statistically different 

(0.56±0.03 and 0.52±0.03, p=0.005), the proportion of 

responders was similar 45%(n=53) and 49%(n=56) p=0.08, 

and there was agreement in 90% of cases (k=0.80). Three 

patients became responders between days 4 and 7 (all died). 

Accuracy of SAH scores to predict 90-day mortality (AUROC) 

was: MELD 0.687 (0.595-0.778), ABIC 0.729 (0.648-0.809), 

LM4 0.686 (0.593-0.775), LM7 0.723 (0.633-0.813). In univariate 

analysis LM4, SIRS, infection, hepatic encephalopathy, 

and gastrointestinal bleeding predicted mortality, however, in 

the multivariate analysis only LM4 (OR=4.67 [1.40-15.54]) 

and GIB (OR=4.94 [1.40-15.25]) remained significant, with a 

trend for HE (OR=1.86 [0.96-4.55]). The operational characteristics 

of LM4 and LM7 as predictors of 90-day mortality are 

shown in table-1. CONCLUSION: LM4 is good as LM7 as predictor 

of 90-day mortality or response to corticosteroids. Using 

LM4 instead of LM7 would help decrease exposure to corticosteroids, 

infection risk, and health care costs. Although there


was a 4% difference in corticosteroid response between LM4 

and LM7, the rate of responders that survived was unchanged 

(36%). Whether earlier responders at day 4 have a favorable 

prognosis need to be investigated. 

Table-1 Operational characteristics of Lille Model on day 4 and 

day 7 

a known inhibitor of autophagy, and AMPK, that activates 

autophagy due to dephosphorylation of these counter regulatory 

signaling molecules. We also observed that the activity of 

protein phosphatase 2A, a critical dephosphorylating enzyme, 

increased while the activity of PI3Kγ, an upstream inhibitor of 

PP2A, was reduced by alcohol. 

Disclosures: 

The following authors have nothing to disclose: Gangarao Davuluri, Samjhana 

Thapaliya, Avinash Kumar, Megan R. McMullen, Rebecca L. McCullough, Laura 

E. Nagy, Sathyamangla V. Naga Prasad, Srinivasan Dasarathy 

Disclosures: 



The following authors have nothing to disclose: Mauricio Garcia-Saenz de 

Sicilia, Chitharanjan Duvoor, Jose Altamirano, Veronica Prado, Juan Caballeria 

1308 

Ethanol-impairs mTOR activity by a novel non-canonical 

PI3Kγ-PP2A axis activating skeletal muscle autophagy 

that underlies sarcopenia. 

Gangarao Davuluri 2 , Samjhana Thapaliya 2 , Avinash Kumar 2 , 

Megan R. McMullen 2 , Rebecca L. McCullough 2 , Laura E. Nagy 2 , 

Sathyamangla V. Naga Prasad 3 , Srinivasan Dasarathy 1 ; 1 Department 

Of Gastroenterology and Hepatology, Cleveland Clinic, 

Cleveland, OH; 2 Pathobiology, Cleveland Clinic, Cleveland, OH; 

3 Molecular Cardiology, Cleveland Clinic, Cleveland, OH 

Background. Sarcopenia or loss of skeletal muscle mass in 

alcoholic cirrhosis is frequent and contributes to morbidity and 

mortality. No therapeutic interventions to reverse sarcopenia 

in alcoholic cirrhosis exist because the mechanisms are poorly 

understood. We have previously observed an alcohol mediated, 

AMPK independent, PI3Kγ dependent inhibition of mTOR 

signaling that activates skeletal muscle autophagy. In the present 

studies we show that ethanol-induced mitochondrial reactive 

oxygen species impairs mTOR activity by a novel non-canonical 

PI3Kγ-protein phosphatase 2 A (PP2A) axis that is responsible 

for increased skeletal muscle autophagy that underlies 

sarcopenia. Methods. Skeletal muscle mass in patients and 

controls and myotube diameter were quantified by image analysis. 

Phosphorylation of AMPK, mTOR and its activity was measured 

by Immunoblots of protein from C2C12 murine myotubes 

exposed to 100 mM ethanol for 6 hr and skeletal muscle from 

mice fed ethanol or pair fed for 25 days. Thin layer chromatography 

blots for PI3Kγ activity was quantified in C2C12 myotubes 

exposed to ethanol and gastrocnemius muscle from mice 

fed ethanol or pair fed controls. Results. Here we report that 

ethanol increases skeletal muscle autophagy in humans, mouse 

models and murine myotubes with unaltered or reduced proteasome 

activity and contributes to sarcopenia. The two major 

regulators of autophagy are mTOR that inhibits autophagy and 

AMPK that activates autophagy. Surprisingly, our data showed 

that both mTOR signaling (autophagy inhibitor) and AMPK 

activation (autophagy activation) were decreased in ethanol 

treated myotubes and in mice exposed to ethanol. The surprising 

signaling response via 2 pathways that converge on autophagy, 

suggested that dephosphorylation may be a potential 

mechanism for reduced phosphorylation of both mTOR and 

AMPK. Consistently, we observed that skeletal muscle PP2A 

activity was increased and was accompanied by a decreased 

PI3Kγ activity, an inhibitor of PP2A. These data showed that 

ethanol activates PP2A dependent dephosphorylation of mTOR 

and AMPK. We confirmed this by using fostriecin, a PP2A 

inhibitor, that reversed ethanol-induced inactivation of mTOR 

and AMPK. Conclusions. Alcohol inhibits activation of mTOR, 

1309 

Differential Changes In Pro-Apoptotic Plasma Glycochenodeoxycholate 

And Fecal Glycodeoxycholate Characterize 

Bile Acid Metabolome In Alcoholic Liver Disease 

Puneet Puri 2 , Andrew R. Joyce 1 , Amon Asgharpour 2 , Mohammad 

S. Siddiqui 2 , Richard K. Sterling 2 , R. Todd Stravitz 2 , Velimir A. 

Luketic 2 , Scott C. Matherly 2 , Kalyani Daita 2 , Susan A. Walker 2 , 

Stephanie Taylor 2 , Christian E. Ammons 2 , Faridoddin Mirshahi 2 , 

Arun J. Sanyal 2 ; 1 Venebio, Richmond, VA; 2 Virginia Commonwealth 

University, Richmond, VA 

BACKGROUND: Bile acid (BA) derangements are implicated 

in the pathophysiology of alcoholic (ALD) and nonalcoholic 

fatty liver disease (NAFLD). Two NAFLD phenotypes, fatty liver 

(NAFL) and steatohepatitis (NASH) relate to disease severity. 

Recently, a BA analogue showed encouraging therapeutic 

potential for NASH. We hypothesized that ALD and NAFLD 

are associated with dysregulated BA metabolism. AIMS: 1) To 

characterize plasma and fecal BA metabolome compared to 

controls (CTL), and 2) To evaluate BA metabolites as potential 

biomarkers of ALD, NAFL and NASH. METHODS: Controls, 

biopsy confirmed NAFL and NASH, and ALD subjects were 

enrolled. Pertinent clinical data, plasma and stool samples 

were collected. High throughput mass spectrometry was used 

for metabolomics. Data were analyzed using pairwise and multiple 

group comparisons by ANOVA, regularized Hotelling’s 

T 2 , and partial least square discriminant analysis with variable 

importance projection (VIP) scores. A VIP score >1 is significant. 

Higher VIP score indicates importance of that variable contributing 

to the change. RESULTS: Four groups with mean(±SD) 

age (yrs): CTL (n=24, 39±12), NAFL (n=22, 53±10), NASH 

(n=37, 58±9) and ALD (n=18, 50±11) were studied. Plasma 

BA metabolome: ALD subjects had markedly increased total 

primary (p


also offer potential for novel biomarkers and therapeutic targets. 

Disclosures: 

Puneet Puri - Advisory Committees or Review Panels: Health Diagnostic Laboratory 

Inc.; Consulting: NPS Pharmaceuticals Inc. 

Andrew R. Joyce - Independent Contractor: Venebio Group, LLC, algorithmRx, 

LLC; Management Position: Venebio Group, LLC, algorithmRx, LLC 

Richard K. Sterling - Advisory Committees or Review Panels: Merck, Vertex, Salix, 

Bayer, BMS, Abbott, Gilead, Baxter, Jansen; Grant/Research Support: Merck, 

Roche/Genentech, Pfizer, Gilead, Boehringer Ingelheim, Bayer, BMS, Abbott 






The following authors have nothing to disclose: Amon Asgharpour, Mohammad 

S. Siddiqui, R. Todd Stravitz, Velimir A. Luketic, Scott C. Matherly, Kalyani Daita, 

Susan A. Walker, Stephanie Taylor, Christian E. Ammons, Faridoddin Mirshahi 

1310 

Ethanol Exposure Inhibits Hepatocellular Lipophagy by 

Inactivating the Small Regulatory GTPase Rab7 

Ryan J. Schulze 1 , Shaun Weller 1 , Micah B. Schott 1 , Karuna 

Rasineni 2 , Barbara Schroeder 1 , Carol A. Casey 2 , Mark A. 

McNiven 1 ; 1 Biochemistry and Molecular Biology, Mayo Clinic, 

Rochester, MN; 2 University of Nebraska Medical Center, Omaha, 

NE 

Alcohol consumption is a well-established risk factor for the 

onset and progression of fatty liver disease, in which an estimated 

90% of heavy drinkers are thought to develop significant 

liver steatosis. For these reasons, an increased understanding 

of the molecular basis for alcohol-induced hepatic steatosis 

is urgently required. Recently, it has become clear that autophagy, 

a catabolic process of intracellular degradation and 

recycling, plays a key role in hepatic lipid metabolism. We 

have previously shown that the small GTPase Rab7, known 

to regulate membrane trafficking and fusion events in the late 

endocytic pathway, also acts as a central orchestrator of hepatocellular 

lipophagy, a selective form of autophagy in which 

lipid droplets (LDs) are specifically targeted for turnover by 

the autophagic machinery (Schroeder et al., 2015 and article 

highlight by Carmona-Gutierrez et al., 2015). Nutrient starvation 

resulted in Rab7 activation on the surface of the LD as 

well as on degradative compartments such as the lysosome. 

This activation resulted in the mobilization of triglycerides 

stored within the LDs for energy production. The GOAL of 

this study was to test whether the steatotic effects of alcohol 

exposure are a result of perturbations in the Rab7-mediated 

lipophagic pathway. Rats fed a 6-week Lieber-DeCarli ethanol-containing 

diet accumulated a significantly higher amount 

of fat in their hepatocytes. Interestingly, primary hepatocytes 

isolated from either these ethanol-fed rats or cultured VA-13 

hepatocytes (HepG2 cells expressing alcohol dehydrogenase) 

contained enlarged and juxtanuclear-localized multivesicular 

bodies (MVBs) and lysosomes that exhibited impaired motility 

and fewer productive interactions with LDs. Importantly, similar 

morphological changes in these organelles are observed 

in hepatocytes subjected to an siRNA-mediated knockdown 

of Rab7. Consistent with these defects in the MVB and lysosomal 

compartments, we observed a marked 80% reduction 

in Rab7 activity in cultured hepatocytes as well as a complete 

block in starvation-induced Rab7 activation in primary hepatocytes 

isolated from chronic ethanol-fed animals. In summary, 

these findings support a direct mechanism whereby acute or 

chronic ethanol exposure inhibits Rab7 activity, resulting in 

the impaired transport, targeting, and fusion of the autophagic 

machinery with LDs, leading to an accumulation of hepatocellular 

lipids and hepatic steatosis. This study was generously supported 

by funding from the NIDDK [5R37DK044650 (MAM)], 

the NIAAA [5R01AA020735 (MAM and CAC)], the Mayo 

Clinic Center for Cell Signaling in Gastroenterology, and the 

Robert and Arlene Kogod Center on Aging. 

Disclosures: 

The following authors have nothing to disclose: Ryan J. Schulze, Shaun Weller, 

Micah B. Schott, Karuna Rasineni, Barbara Schroeder, Carol A. Casey, Mark 

A. McNiven 

1311 

Alcohol modifies the composition of the intrahepatic 

macrophage pool through FOXO3-dependent and cell 

type specific apoptosis 

Zhuan Li, Jie Zhao, Steven A. Weinman; Department of Internal 

Medicine, University of Kansas Medical Center, Kansas City, KS 

Hepatic macrophages are a heterogeneous population consisting 

of both resident and infiltrating macrophages that carry 

out diverse pro- and anti-inflammatory functions necessary 

for homeostasis, disease progression and injury repair. The 

mechanisms that regulate macrophage subsets in alcoholic 

liver disease are not yet understand. We recently showed that 

FOXO3 protects the liver from alcohol-induced inflammation. 

It also induces S574 phosphorylation of FOXO3 which selectively 

enhances expression of pro-apoptotic genes and induces 

apoptosis. FOXO3-dependent apoptosis was induced in macrophages 

by LPS. The AIMS of this study were to determine 

whether FOXO3-dependent macrophage apoptosis protects 

the liver from alcohol through changes in intrahepatic macrophage 

populations. METHODS: ChIP assays assessed promoter 

binding. qPCR was used to measure gene expression. Mouse 

peritoneal macrophages were treated with LPS for 24 hours or 

differentiated with INF-γ or IL-4 before LPS treatment. Mice were 

gavaged once with alcohol or fed a Lieber-DiCarli alcohol diet 

for 3 wks. RESULTS: LPS treatment of THP-1 monocytes induced 

FOXO3 S574 phosphorylation, enhanced expression of TRAIL, 

suppressed Bcl-2 and induced apoptosis. A similar result was 

obtained with peritoneal macrophages from wild-type (wt) 

mice but none of these effects occurred in FOXO3 -/- macrophages. 

To determine if apoptosis was selective for macrophage 

type, we separately measured LPS-induced apoptosis in 

INF-γ differentiated macrophages (M1) and IL-4 differentiated 

macrophages (M2). Only M1 macrophages underwent apoptosis 

and this was FOXO3 dependent. Acute alcohol gavage 

resulted in nearly a 50% decrease of hepatic macrophage 

number by 9 hours in wt but not FOXO3 -/- mice and this loss 

of macrophages was selective for cells with M1 markers. 

Unlike acute gavage, chronic ethanol feeding had no effect 

on hepatic macrophage number or M1/M2 ratio in wt mice 

but increased the M1/M2 ratio by 50% in FOXO3 -/- mice. 

In the FOXO3 -/- mice there was a strong positive correlation 

between M1 pro-inframammary macrophage number and ALT 

(P


The following authors have nothing to disclose: Zhuan Li, Jie Zhao 

1312 

PDE4 inhibition attenuates alcohol induced hepatic oxidative 

stress by increasing antioxidant enzyme expression 

Diana Avila 1 , Jingwen Zhang 1 , Craig J. McClain 1,2 , Shirish 

Barve 1 , Leila Gobejishvili 1 ; 1 Department of Medicine/GI, University 

of Louisville, Louisville, KY; 2 Robley Rex VAMC, Louisville, KY 

Alcohol metabolism leads to generation of free radicals and 

oxidative stress with a resultant formation of lipid peroxidation 

products, which contribute to the development of alcoholic 

liver disease (ALD). Additionally, alcohol decreases antioxidant 

capacity of liver, specifically nuclear factor erythroid 2– 

related factor 2 (Nrf2) dependent enzymes. cAMP increasing 

agents have been shown to effectively mitigate oxidative stress 

both in vivo and in vitro. Our previous work has shown that 

alcohol increases the expression of PDE4, cAMP degrading 

enzyme, in macrophages and hepatic Kupffer cells. Hence, 

we hypothesized that compromised cAMP levels due to alcohol 

might contribute to the decreased antioxidant capacity 

of liver, and restoring cAMP signaling could prevent alcohol 

induced oxidative stress. To test this hypothesis, we employed 

two approaches to block degradation of cAMP by phosphodiesterases 

(PDEs): genetic (by using PDE4B knockout mice) 

and pharmacological inhibition of PDE4 (Rolipram) in a mouse 

model of ALD. C57BL/6 and Pde4b knockout (pde4b -/- ) male 

mice were pair-fed Lieber-DeCarli liquid diet containing either 

alcohol (AF) or isocaloric maltose dextrin (PF) for 4 weeks. 

Additional groups of mice were treated with Rolipram at 5 

mg/kg, 3 times a week for 4 weeks. Mice were sacrificed 

at 1, 2 and 4 weeks after starting alcohol. Liver cAMP levels 

were measured by cAMP ELISA kit; Hepatic inflammation and 

oxidative stress was examined by immunohistochemical staining 

of liver tissue with 4HNE (4-Hydroxynonenal) and F4/80 

(monocyte/macrophage marker) antibodies. Effect of alcohol 

on activation of Nrf2 was evaluated by examining nuclear 

Nrf2 levels. Further, expression of Nrf2-dependent hepatic antioxidant 

enzymes, superoxide dismutase 1 and 2 (SOD1/2), 

Catalase and glutathione peroxidase (GPx) was analyzed by 

Western blot. Alcohol feeding resulted in a significant increase 

in hepatic PDE4 expression and a decrease in cAMP levels. 

Liver 4HNE and F4/80 staining was significantly increased in 

alcohol fed WT mice demonstrating increased oxidative stress, 

but was significantly attenuated by PDE4 inhibition. Alcohol 

reduced Nrf2 activation in wild type mice but not in Rolipram 

treated and PDE4B knockout mice. Catalase and SOD2 levels 

did not seem to be affected by alcohol, but SOD1 and GPx1/2 

were decreased in alcohol fed wild type mice. Notably, in comparison 

with wild type, Pde4b knockout and Rolipram treated 

alcohol fed mice had higher levels of SOD1/2, and GPx1/2. 

In summary, these data indicate that alcohol effect on hepatic 

cAMP levels contribute to compromised antioxidant capacity of 

the liver, which could be restored by PDE4 inhibition. 

Disclosures: 





The following authors have nothing to disclose: Diana Avila, Jingwen Zhang, 

Leila Gobejishvili 

1313 

Hepatocyte-specific deletion of the essential circadian 

clock transcription factor BMAL1 enhances chronic ethanol-induced 

depletion of hepatic glycogen in mice 

Uduak S. Udoh, Telisha Swain, Shannon Bailey; Pathology, University 

of Alabama at Birmingham, Birmingham, AL 

The molecular circadian clock plays a central role in regulating 

fundamental physiological and metabolic processes and allows 

organisms/organs/cells to anticipate and adapt to changes in 

their 24 hr environment. Previously, we found that chronic alcohol 

consumption disrupts the liver molecular clock mechanism 

and daily rhythms in liver glycogen metabolism. Herein, our 

goal was to determine the effect chronic alcohol consumption 

has on hepatic glycogen metabolism using a model of hepatocyte 

clock dysfunction; i.e., the hepatocyte-specific BMAL1 

knockout (HBK) mouse. For these studies, single-housed male 

HBK and wild type (WT) littermates were kept under a 12:12 

hr light-dark cycle and fed either the Lieber-DeCarli control 

diet or the ethanol-containing (3% w/v) diet using a pair-fed 

isocaloric study design for 5 weeks. Livers were collected every 

4 hr for a 24 hr period at Zeitgeber time (ZT) 3, 7, 11, 15, 

19, and 23 where ZT 0 = lights on (beginning of less active 

phase) and ZT 12 = lights off (beginning of more active phase), 

and used for glycogen determination and gene expression. 

Chronic alcohol consumption significantly decreased glycogen 

content in livers of WT mice at all times of the day. The diurnal 

rhythm in glycogen was also altered in livers of WT ethanol-fed 

mice as compared to the rhythm measured in livers of WT control-fed 

mice [Mesor/Mean: WT-Control (40.30) vs. WT-Ethanol 

(24.87), p


inhibition of two ceramide synthetic pathways on hepatic Plin2 

expression, steatosis, and glucose and lipid homeostasis in 

vivo. In vitro, we investigated the individual effects of de novo 

synthetic enzymes on Plin2 regulation. METHODS Mice were 

fed a Lieber-DeCarli control or ethanol diet for 4 weeks and 

pharmacologically treated i.p. with imipramine (an inhibitor 

of sphingomyelinase (SMASE)); myriocin (an inhibitor of serine 

palmitoyl transferase (SPT), the rate limiting enzyme of de 

novo ceramide synthesis); or saline. Various metabolic parameters, 

serum chemistries and glucose tolerance were measured. 

Liver samples were taken for immunoblotting, mRNA analysis 

and ceramide measurement by mass spectrometry. In vitro, we 

incubated VL-17A cells (ethanol-metabolizing human hepatoma 

cells) with control or ethanol media and inhibited key enzymes 

in the de novo ceramide synthetic pathway with myriocin, 

fumonisin B1 (FB-1, an inhibitor of ceramide synthase), or 

GT-11 (an inhibitor of dihydroceramide desaturase). Plin2 

mRNA and protein, lipids, and markers of translation were 

quantified. RESULTS SPT and SMASE inhibition in alcohol-fed 

mice reduced hepatic triglycerides (TG) by approximately 37% 

(p


BMI 29, 44% coffee users, 13% black tea users, 4% green tea 

users, mean ALT 26, mean AST 27, mean bilirubin 0.5 mg/ 

dL, mean albumin 3.8 mg/dL) were recruited. Apart from liver 

tests, cases and controls differed for age (P=0.046), education 

level (P=0.03), coffee use (P=0.0002). On a logistic regression 

analysis model after controlling for relevant co-variates, 

white race was associated with AH with OR (95% CI) of 5.5 

(1.8-16.5). In contrast, odds of AH were lower by about 80% 

among coffee users; 0.21 (0.10-0.45). Recidivism was self-reported 

by 41 (16 cases) and 14 (6 cases) at 6 and 12 months 

respectively. On a logistic regression analysis model, low (


intestinal crypts that are enriched in Paneth cells resulted in a 

dose- and time-dependent increase in IL-17 protein production. 

Conclusions: Our results highlight substantial hepatic immune 

cell changes due to alcohol and indicate that increased number 

of Paneth cells in the small intestine is an early event in alcohol-induced 

intestinal pathology. For the first time, we describe 

that IL-17 production by Paneth cells is induced by alcohol in 

small intestinal crypts and represents an early intestinal inflammatory 

marker in ALD. These observations identify a novel role 

of Paneth cells and intestinal IL-17 in ALD and highlight the 

importance of the gut-liver axis in alcoholic liver disease. 

Disclosures: 

The following authors have nothing to disclose: Benedek Gyongyosi, Patrick 

Lowe, Arvin Iracheta-Vellve, Abhishek Satishchandran, Aditya Ambade, Banishree 

Saha, Gyongyi Szabo 

1319 

Microbiota protects mice against acute alcohol-induced 

liver injury 

Peng Chen, Yukiko Miyamoto, Magdalena Mazagova, Lars Eckmann, 

Bernd Schnabl; University of California San Diego, La Jolla, 

CA 

Background: Chronic alcohol abuse is associated with intestinal 

bacterial overgrowth, increased intestinal permeability, 

and translocation of microbial products from the intestine to 

the portal circulation and liver. There is strong evidence that 

experimental alcoholic liver disease depends on translocated 

microbial products. The aim of our study was to investigate 

the physiological relevance of the intestinal microbiota in 

alcohol-induced liver injury using conventional and germ-free 

C57BL/6 mice. Methods: We employed germ-free mice in a 

model of acute alcohol exposure that mimics binge drinking. 

Acute alcoholic liver injury was induced by a single gavage of 

300 μl of 30% (vol/vol) ethanol. Control mice were gavaged 

with an isocaloric amount of dextrose (300 μl, 0.42 g/ml). 

Mice were harvested 9 hours after gavage. Results: Germ-free 

mice showed significantly greater liver injury as determined 

by plasma ALT levels and H&E staining of liver sections after 

oral gavage of ethanol compared with conventional mice. 

This was accompanied by increased hepatic expression of 

macrophage and neutrophil markers. Several proinflammatory 

cytokines and chemokines including TNFα, Ccl2, Ccl4, Ccl5, 

Cxcl1, Cxcl2 and Cxcl5 were increased in alcohol-binged 

germ-free relative to conventional mice. In parallel, germ-free 

mice exhibited increased hepatic steatosis and upregulated 

expression of genes involved in fatty acid and triglyceride synthesis 

compared with conventional mice after acute ethanol 

administration. The absence of microbiota was also associated 

with increased hepatic expression of ethanol metabolizing 

enzymes, which led to faster ethanol elimination from the 

blood and lower plasma ethanol concentrations. In particular, 

hepatic Cyp2e1 gene and protein expression were significantly 

higher in germ-free mice before and after an alcohol 

binge. Furthermore, expression of aldehyde dehydrogenase 2 

(Aldh2), which catalyzes the transformation of acetaldehyde 

to acetic acid, was significantly higher in control and alcohol 

challenged germ-free mice. Intestinal levels of ethanol metabolizing 

genes showed regional expression differences, and 

were overall higher in germ-free relative to conventional mice. 

In conclusion, our findings indicate that absence of the intestinal 

microbiota increases hepatic ethanol metabolism and the 

susceptibility to binge-like alcohol drinking. The commensal 

microbiota has therefore a beneficial role in protection against 

acute alcoholic liver disease. 

Disclosures: 

The following authors have nothing to disclose: Peng Chen, Yukiko Miyamoto, 

Magdalena Mazagova, Lars Eckmann, Bernd Schnabl 

1320 

Hepatocyte-specific StARD1 deletion attenuates alcoholic 


Vicent Ribas 1,2 , Susana Nuñez 2,1 , Carmen Garcia-Ruiz 2,1 , Jose 

Fernandez-Checa 2,1 ; 1 Liver Unit, Hospital Clínic i Provincial de 

Barcelona, IDIBAPS, Barcelona, Spain; 2 Instituto de Investigaciones 

Biomédicas de Barcelona (IIBB-CSIC)), Barcelona, Spain 

Previous studies have shown that chronic alcohol feeding 

results in the depletion of mitochondrial GSH due to the accumulation 

of cholesterol in mitochondria. Mitochondrial GSH 

depletion stimulates reactive oxygen species and oxidative 

stress, and sensitizes hepatocytes to hypoxia and inflammatory 

cytokines-mediated apoptosis. Steroidogenic acute regulatory 

domain 1 (StARD1) protein was identified in steroidogenic 

tissues as a key protein responsible for cholesterol trafficking 

to the mitochondrial inner membrane for the synthesis of steroids. 

Mice with global StARD1 deletion die within 10 days 

due to adrenal lipoid hyperplasia. To circumvent this limitation, 

we generated mice with hepatocyte-specific deletion of 

StARD1 to study its role in alcoholic steatohepatitis induced 

by a novel model in which the Lieber DeCarli ethanol liquid 

diet was supplemented with 0.2% cholesterol. METHODS: We 

generated mice with floxP sites flanking exons 2-5 of StARD1 

(floxed mice) and were crossed with Albumin-Cre transgenic 

mice. StARD1 liver-specific knockout animals (SLKO) and their 

corresponding StARD1 floxed control littermates were fed a 

Lieber DeCarli ethanol liquid diet supplemented with 0.2% cholesterol 

(L-C+Ch) for four weeks. At the end of this period, we 

determined parameters of liver damage, mitochondrial cholesterol 

and GSH homeostasis, markers of endoplasmic reticulum 

(ER) stress, inflammation, fibrosis and expression of lipogenic 

genes. RESULTS: In agreement with previous studies, StARD1 

mRNA was induced by L-C+Ch feeding in StARD1 floxed 

animals (2-4 fold) while StARD1 expression was effectively 

ablated in SLKO liver. In parallel, L-C+Ch diet increased serum 

ALT (200-250U/L) in StARD1 floxed animals, an event that 

was significantly reduced by 50-60% in SLKO animals. Interestingly, 

total liver cholesterol was increased by L-C+Ch treatment 

in both genotypes, but mitochondrial cholesterol loading 

was increased only in StARD1 floxed animals. In line with 

this, SLKO liver mitochondria exhibited increased mGSH levels 

after L-C+Ch consumption, when compared to StARD1 floxed 

mice. Additionally, L-C+Ch feeding increased expression of ER 

stress markers (Chop, Atf6, Pdi), genes involved in lipogenesis 

(Srebp1, Srebp2, Dgat2, Fasn), fibrosis (Col1a1, Acta2) and 

inflammation (Ccl2, Tnfa, IL-6 and IL-1b) in StARD1 floxed mice 

(3-6 fold), whereas the rise in these markers was markedly 

attenuated (40-70%) in SLKO mice. CONCLUSIONS: These 

results suggest a key role of StARD1-mediated mitochondrial 

cholesterol trafficking in the pathogenesis of alcoholic steatohepatitis, 

indicating that StARD1 may be a novel target for the 

treatment of alcohol induced liver disease. 

Disclosures: 

The following authors have nothing to disclose: Vicent Ribas, Susana Nuñez, 

Carmen Garcia-Ruiz, Jose Fernandez-Checa


1321 

Infection in alcoholic hepatitis: a major prognostic indicator 

and potential therapeutic target 

Richard Parker 1,4 , Fiona Jones 2 , Daniel J. Wheatley 3 , Ian A. 

Rowe 1 , Christopher Corbett 3 , Andrew Holt 4 , Stephen Stewart 2 ; 

1 Centre for Liver Research, University of Birmingham, Birmingham, 

United Kingdom; 2 Mater Misericordiae University Hospital, Dublin, 

Ireland; 3 New Cross Hospital, Wolverhampton, United Kingdom; 

4 University Hospitals Birmingham NHS Foundation Trust, Birmingham, 


Introduction We sought to detail the incidence, consequences 

and causes of infection in a cohort of patients with Alcoholic 

Hepatitis (AH). Methods Three centres in the UK and Ireland 

gathered data from patients with a clinical diagnosis of alcoholic 

hepatitis. Clinical, biochemical and microbiological 

records were reviewed retrospectively. Infection was defined 

by evidence of consolidation on chest x-ray, positive urinalysis, 

ascitic fluid analysis showing 250 neutrophils/mm 3 or 

positive microbiological culture. Survival was analysed with 

Kaplan-Meier (KM) curves and Cox proportional hazard analysis 

to control for confounding factors. Characteristics of patients 

with or without evidence of infection were compared with twotailed 

student’s t-test. Results In total 215 patients were included 

with an median follow-up period of 30 months. Average discriminant 

function (DF) was 72.7 and 85% of patients had 

severe disease (DF >32). Evidence of infection was seen in 97 

patients (45%). Patients with infection had significantly higher 

serum urea (p


1323 

Ethanol-Induced Alterations In The Intestinal Methionine 

Metabolic Pathway Promote Tight Junction Disruption 

Dan Feng 1,2 , Paul G. Thomes 3 , Natalia A. Osna 1,2 , Dean J. 

Tuma 1,2 , Kusum K. Kharbanda 1,2 ; 1 Research Service, Veterans 

Affairs Nebraska-Western Iowa Health Care System, Omaha, NE; 

2 Department of Internal Medicine, University of Nebraska Medical 

Center, Omaha, NE; 3 Liver Pathobiology Laboratory, Cannon 

Research Center, Carolinas Healthcare System, Charlotte, NC 

The gut-liver interaction has emerged as a critical component 

in the pathogenesis of alcoholic liver disease (ALD). The central 

mediator of ALD progression is the gut luminal antigens, especially 

endotoxins that translocate to the liver via compromised 

gut epithelial tight junctions (TJ). We have previously shown 

that ethanol consumption can alter the liver methionine metabolic 

pathway to impair several methylation reactions leading 

to the generation of many hallmark features of alcoholic liver 

injury. This study was undertaken to examine whether alcohol 

exposure alters the intestinal methionine metabolic pathway 

and to explore whether these alterations play a causal role in 

disrupting the gut barrier integrity. Adult male Wistar rats were 

pair-fed the Lieber DeCarli control or ethanol diet for 8 weeks. 

At the end of the feeding regimen, intestines were removed and 

analyzed. We observed the level of methionine synthase (MS, 

an enzyme of the methionine metabolic pathway that prevents 

buildup of a harmful metabolite, S-adenosylhomocysteine, 

SAH) was decreased by 50% in the ileum of ethanol-fed rats 

compared to controls. This decrease was accompanied by a 

compensatory increase in betaine homocysteine methyltransferase 

(BHMT), another enzyme that maintains the level of a vital 

methylating agent, S-adenosylmethionine (SAM), and prevents 

SAH buildup. However, while the adaptive increase in BHMT 

maintained SAM levels in the ileum of the ethanol-fed rat, there 

was a significant elevation in SAH levels ultimately lowering 

the SAM:SAH ratio compared to controls. All these changes 

were similar to those previously reported in the livers of ethanol-fed 

rats. Concomitant with the changes in the crucial components 

of the methionine metabolic pathway that controls the 

cellular methylation potential, we saw the disassembly of TJs 

in the ileum of the ethanol-fed rats. There was a disorganized 

localization of the TJ proteins, occludin and claudin -1, which 

normally seal the intercellular spaces to prevent the paracellular 

translocation of luminal antigens into portal circulation. Further 

studies using an inhibitor of lysine methyltransferase identified 

that the methylation reactions catalyzed by this enzyme 

were important for maintaining the barrier integrity. We further 

observed that occludin is methylated and this post-translational 

modification occurred on the lysine and not arginine residue(s). 

Taken together, our results indicate that alcohol-induced alterations 

in the intestinal methionine metabolic pathway and the 

resulting impairments in methylation reactions contribute to the 

disruption of the TJs, thereby promoting gut leakiness and progressive 

liver injury. 

Disclosures: 

The following authors have nothing to disclose: Dan Feng, Paul G. Thomes, 

Natalia A. Osna, Dean J. Tuma, Kusum K. Kharbanda 

1324 

Role of Pre-mRNA Splicing Regulator SLU7 in Mediating 

Detrimental Action of Ethanol in the Liver 

Jiayou Wang 1,2 , Xudong Hu 1,3 , Alvin Jogasuria 1 , Kwangwon Lee 1 , 

Jiashin Wu 1 , Min You 1 ; 1 Pharmaceutical Sciences, Northeast Ohio 

Medical University (NEOMED), Rootstown, OH; 2 Department of 

Anatomy, Guangzhou University of Chinese Medicine, Guangzhou, 

China; 3 Department of Biology, Shanghai University of Traditional 

Chinese Medicine, Shanghai, China 

Precursor mRNA (pre-mRNA) splicing is a critical step in gene 

expression that removes intronic sequences from immature 

mRNA, producing mature mRNA that can be translated into 

protein. Errors in splicing contribute to at least 15% of human 

genetic disorders. Aberrant pre-mRNA splicing plays a significant 

role in liver diseases, but very little is known about its 

role in alcoholic steatosis/steatohepatits. Here, we identified 

a new target of ethanol action in the liver, namely, pre-mRNA 

splicing regulator SLU7. The effects of ethanol on splicing factor 

SLU7 were assessed in cultured hepatocytes and in animal 

liver. Feeding mice a modified Lieber-DeCarli ethanol-containing 

liquid diet for 4-wks resulted in significant decreases in the 

mRNA and protein expression levels of hepatic SLU7. More 

importantly, ethanol exposure robustly reduced the amount of 

splicing factor SLU7 in the nucleus in cultured hepatocytes and 

in mouse liver. Slu7 governs the splicing and/or expression of 

multiple genes essential for hepatic lipid metabolism, including 

serine/arginine-rich splicing factor 3 (SRSF3) and hepatocyte 

nuclear factor 4α (HNF4α). SRSF3 pre-mRNA alternative splicing 

generates the full-length isoform lacking exon 4 (Iso1) and 

an alternative isoform including exon 4 (Iso2). Ethanol-mediated 

disruption of splicing factor SLU7 triggered altered SRSF3 

pre-mRNA splicing by inhibiting the mRNAs of total SRSF3, 

SRSF3_Iso 1, or SRSF3_ Iso2 and HNF-4α in the livers of ethanol-fed 

mice. Remarkably, the alteration of hepatic SLU7-SRSF3 

axis by ethanol exposure was closely associated with the development 

of excess accumulation of hepatic triglycerides in mice. 

Taken together, our findings suggest that splicing regulator 

SLU7 may play an essential role in regulating the effects of ethanol 

on hepatic lipid metabolism and development of alcoholic 

fatty liver. 

Disclosures: 

The following authors have nothing to disclose: Jiayou Wang, Xudong Hu, Alvin 

Jogasuria, Kwangwon Lee, Jiashin Wu, Min You 

1325 

Impaired Proteasome Function is Associated with 

Alcohol-Induced Hepatic Receptor Interacting Protein 

3-Mediated Necroptosis, Steatosis and Liver Injury 

Wen-Xing Ding, Shaogui Wang, Hong-Min Ni; Pharmacology, 

Toxicology and Therapeutics, The University of Kansas Medical 

Center, Kansas City, KS 

Both hepatocyte apoptosis and necrosis are hallmarks of alcohol-induced 

liver injury. Recent evidence suggests that receptor-interacting 

protein kinase (RIP) 3-mediated necroptosis is 

important in chronic alcohol feeding-induced liver injury in 

mice. However, the mechanisms by which alcohol activates 

RIP3-mediated necroptosis are unclear. Using the recently established 

chronic alcohol feeding plus binge (Gao-binge) model, 

we demonstrated that Gao-binge alcohol treatment increased 

RIP3 but not RIP1 protein levels in mouse livers based on both 

western blot analysis and immunohistochemical staining for 

RIP1 and RIP3. 7-OCl-Nec 1, a recently developed specific 

RIP1 inhibitor, failed to protect against Gao-binge alcohol-induced 

steatosis and liver injury, suggesting alcohol-induced


necroptosis is RIP1-independent. In contrast, RIP3 knockout 

mice had decreased neutrophil infiltration, serum alanine 

amino transferase (ALT) activity and steatosis compared to wild 

type mice after Gao-binge alcohol treatment, suggesting that 

RIP3 but not RIP1 contributes to alcohol-induced necroptosis 

and liver injury. Gao-binge alcohol treatment did not change 

hepatic mRNA levels of RIP3 but decreased mRNA levels of 

RIP1, suggesting the increase in hepatic RIP3 protein level in 

alcohol-treated mouse livers was likely due to post-translational 

regulation. In primary cultured mouse and human hepatocytes, 

we found that alcohol treatment decreased proteasome activity 

and increased the protein level of RIP3. Similarly, Bortezomib, 

a proteasome inhibitor, also increased the protein level of RIP3 

in cultured hepatocytes. Consistent with these in vitro findings, 

we also found that Gao-binge alcohol treatment decreased 

the levels of proteasome subunit alpha type-5 (PSMA5) and 

proteasome subunit beta type-5 (PSMB5), two key proteasome 

subunits that are important for proteasome function. As a result, 

Gao-binge treatment also decreased proteasome activity in 

the mouse livers. More importantly, we also found decreased 

expression of PSMA5 and PSMB5 and increased protein levels 

of RIP3 in human alcoholic liver biopsy samples compared 

to healthy human livers. In conclusion, results from this study 

suggest that impaired hepatic proteasome function by alcohol 

exposure may contribute to alcohol-induced steatosis and liver 

injury by inducing necroptosis through blocking proteasomal 

degradation of RIP3. 

Disclosures: 

The following authors have nothing to disclose: Wen-Xing Ding, Shaogui Wang, 

Hong-Min Ni 

1326 

Characteristic features of microbiomes in alcoholic 

liver disease analyzed by 16S ribosomal RNA gene 

pyrosequencing and transcriptional fragment length 

polymorphism methods using fecal samples: the impact 

of abstinence 

Makiko Taniai, Etsuko Hashimoto, Kuniko Yamamoto, Yuichi 

Ikarashi, Kazuhisa Kodama, Tomomi Kogiso, Nobuyuki Torii, Katsutoshi 

Tokushige; Internal Medicine, Institute of Gastroenterology, 

Tokyo Women’s Medical University, Tokyo, Japan 

Intestinal bacterial overgrowth and dysbiosis 

induced by ethanol ingestion appears to play an important 

role in the pathogenesis of alcoholic liver disease (ALD). 

Recently, determination of microbiomes by 16S ribosomal RNA 

(rRNA) gene pyrosequencing and terminal restriction fragment 

length polymorphism (TRFLP) using stool samples has been 

widely used in place of conventional culture methods for the 

assessment of the diversity of complex bacterial communities 

and rapid comparison of the community structure. In this study, 

we evaluated the characteristics of intestinal microbiomes in 

ALD patients and the impact of abstinence on it using TRFLP 

analysis. We investigated samples from 20 patients 

clinicopathologically diagnosed as ALD (75% males, 21 to 

76 years, including 7 patients with cirrhosis) and 10 healthy 

volunteers as controls matched with age and sex. Five mg of 

feces were collected for analysis. Principle component analysis 

(PCA) and phylogenetic cluster analysis were employed to 

assess the comparison about the component of microbiomes 

in two groups. In 6 cases who successfully stopped drinking, 

we rechecked the stool samples at the point from 3 to 6 weeks 

after abstinence. Large differences at microbial phylum, 

family, and genus levels were noted between the patients 

with ALD and control subjects. The most striking feature about 

microbiomes in ALD patients was the variability of microbiomes 

were significantly decreased and very simplified compared to 

that of control subjects. In ALD patients, the increase of Bacteroides 

fragilis group and Clostridium coccoides group and the 

decrease of Lactobacillus group were prominent. Interestingly, 

in all cases who stopped drinking, the variability of microbiomes 

dramatically improved and the components approached 

to those of controls. Intestinal microbiomes 

in ALD patients showed distinct composition and significant 

decrease in the variability of microbiomes compared to that of 

control subjects. Those dysbiosis improved after relatively shortterm 

abstinence. This findings could lead to the elucidation of 

pathogenesis and new therapeutic approach for ALD. 

Disclosures: 

The following authors have nothing to disclose: Makiko Taniai, Etsuko Hashimoto, 

Kuniko Yamamoto, Yuichi Ikarashi, Kazuhisa Kodama, Tomomi Kogiso, 

Nobuyuki Torii, Katsutoshi Tokushige 

1327 

LPS-TLR4 pathway mediates ductular reaction expansion 

in alcoholic hepatitis 

Gemma Odena 1 , Raluca Dumitru 2 , Jiegen Chen 1 , Jose Altamirano 

3,4 , Veronica L. Massey 1 , Hiroshi Matsushita 5 , Daniel Rodrigo-Torres 

3 , Oriol Morales-Ibanez 3 , Juan Caballeria 6,3 , Pere 

Gines 6,3 , Ekihiro Seki 5 , Pau Sancho-Bru 3 , Ramon Bataller 1,3 ; 

1 Division of Gastroenterology and Hepatology, Departments of 

Medicine and Nutrition and Bowles Center For Alcohol Studies, 

University of North Carolina at Chapel Hill, Chapel Hill, NC; 

2 UNC Human Pluripotent Stem Cell Core, Neuroscience Center 

and Department of Genetics, University of North Carolina at 

Chapel Hill, Chapel Hill, NC; 3 Institut d’Investigacions Biomediques 

August Pi i Sunyer (IDIBAPS), CIBER de Enfermedades Hepaticas y 

Digestivas (CIBERehd), Barcelona, Spain; 4 Liver Unit-Internal Medicine 

Department, Vall d’Hebron University Hospital, Vall d’Hebron 

Institut de Recerca, Barcelona, Spain; 5 Division of Gastroenterology, 

Department of Medicine, Cedars-Sinai Medical Center, Los 

Angeles, CA; 6 Liver Unit, Hospital Clínic, Barcelona, Spain 

Alcoholic hepatitis (AH) is the most severe form of alcoholic 

liver disease and targeted therapies are urgently needed. We 

recently showed that severe AH is characterized by progenitor 

cell expansion and inefficient hepatic regeneration. Because 

LPS is a major molecular driver in AH and its serum levels 

correlate with patient outcome, we investigated whether the 

LPS-TLR4 pathway mediates progenitor cell expansion. We first 

assessed the potential role of LPS in patients with biopsy-proven 

AH (n=28). LPS serum levels correlated with disease severity 

(ABIC score; r=0.48, p=0.02) and markers of progenitor cell 

expansion (KRT23, r=0.58, p


ulation of LPC markers gene expression (EpCam, KRT7 and 

KRT23). Importantly, LPS stimulated gene and protein expression 

of LPCs markers. Interestingly, human LPCs also showed 

a profibrogenic profile, as indicated by increased expression 

of collagen and α-smooth muscle actin (7.3 and 3.2-fold, 

respectively; p5 MTs among parents was observed in 219 (21.7%), 305 

(30.3%) and 109 (10.8%) cases, respectively. 59 (5.8%) and 

261 (25.9%) cases had a history of cirrhosis and alcoholism 

among parents, respectively. Alcoholic cirrhotics with FH of 

MTs had significantly younger age at diagnosis (42.8±9.3 

vs 48.3±10.4 years, p


high utilization of governmental programs. More effective early 

identification, prevention and treatment of ETOH liver disease 

are needed. 

Disclosures: 

Rolland C. Dickson - Advisory Committees or Review Panels: Biotest; Speaking 

and Teaching: gilead 

The following authors have nothing to disclose: Jennifer Saad, Neil Volk, Shawn 

Shah, Joseph Shatzel, Harley Friedman 

1330 

Gut Bacterial Load Influences Neutrophil Infiltration 

And Liver Inflammation In An Acute-On-Chronic Alcohol 

Feeding Model In Mice 

Patrick Lowe, Benedek Gyongyosi, Abhishek Satishchandran, 

Arvin Iracheta-Vellve, Aditya Ambade, Gyongyi Szabo; Medicine, 

UMass Medical School, Worcester, MA 

Background/Aims: The triggers for alcoholic hepatitis involve 

both metabolic and inflammatory danger signals. Previous studies 

showed that gut-derived LPS activates Kupffer cells, but the 

effect of the gut microbiome on neutrophil infiltration, a major 

factor in alcoholic hepatitis, is not known. In this study, we 

investigated the role of gut bacterial load on alcohol-induced 

liver inflammation and immune cell infiltration. Methods: 6-8 

week-old C57BL/6 female mice received 10 days of 5% ethanol 

in liquid diet (Lieber DeCarli) followed by one ethanol 

gavage (5g ethanol/kg body weight) or pair-fed diet followed 

by sugar gavage. Some mice received an antibiotic cocktail 

(ampicillin, neomycin, metronidazole and vancomycin) twice 

daily orally five days prior to and during alcohol consumption. 

Mice were sacrificed 9 hours after the alcohol gavage. 

Results: The alcohol-induced increase in gut bacterial load was 

reduced 18-fold in antibiotic-treated mice as measured by bacterial 

culture of cecum stool samples. Alcohol feeding increased 

liver TNFα (4-fold), MCP-1 (8-fold) and CXCL1 (15-fold) mRNA 

compared to control diet and gut bacterial reduction eliminated 

these increases. Serum MCP-1 protein was increased in alcohol-fed 

mice (1409 pg/mL compared to 818 pg/mL in pairfed) 

while antibiotic-treated alcohol-fed animals showed no 

elevation in serum MCP-1 (689 pg/mL) compared to pair-fed 

animals. Further, we found elevated E-selectin mRNA and MPO 

immunohistochemical staining, indicators of neutrophil activation, 

in livers of alcohol-fed compared to control mice and 

these neutrophil markers were significantly reduced in antibiotic-treated 

alcohol-fed mice. This suggests a reduction in liver 

neutrophil infiltration and activation upon antibiotic treatment. 

Unlike inflammation, liver injury was not dependent on gut 

bacterial load as ALT levels were elevated in alcohol-fed mice 

compared to pair-fed controls with and without antibiotic treatment 

(177 and 190 IU/L, respectively). However, antibiotics 

did provide protection from lipid accumulation as alcohol-fed 

antibiotic treated mice had decreased liver Oil-Red O staining 

compared with non-treated alcohol-fed mice. Conclusions: Our 

results suggest that gut bacterial load correlates with the extent 

of liver inflammation, neutrophil infiltration, systemic cytokine 

increase and steatosis following alcohol consumption, while 

alcohol-induced liver damage (increased ALT) is independent 

of gut bacterial load. These observations highlight the importance 

of gut microbiome in the dynamics of neutrophil infiltration 

and indicate a role of gut- and pathogen-derived signals in 

neutrophil activation in alcoholic hepatitis. 

Disclosures: 

The following authors have nothing to disclose: Patrick Lowe, Benedek Gyongyosi, 

Abhishek Satishchandran, Arvin Iracheta-Vellve, Aditya Ambade, Gyongyi 

Szabo 

1331 

Tributyrin, a butyrate pro-drug, attenuates alcohol-induced 

inflammation and neutrophil-mediated hepatic 

injury 

Hridgandh Donde 1 , Smita Ghare 1 , Jingwen Zhang 1 , Swati Joshi- 

Barve 1 , Craig J. McClain 1,2 , Shirish Barve 1 ; 1 Department of Medicine/GI, 



Background: Emerging evidence has shown that neutrophils 

and hepatic inflammation are major drivers of alcoholic liver 

disease (ALD). Neutrophils infiltration is critically involved in 

the liver dysfunction and cellular injury in ALD. Hence, attenuation 

of these may prove to be beneficial in the treatment of ALD. 

Our recent work indicated that chronic alcohol consumption 

decreased intestinal short chain fatty acids, particularly butyrate, 

which has a major impact on the development of intestinal 

barrier dysfunction and ALD. The aim of this study was to investigate 

whether oral administration of tributyrin (Tb; a butyrate 

prodrug) results in protection against ALD by targeting alcohol 

induced inflammation and its effects in the liver. Tb is a triglyceride 

that is rapidly absorbed and metabolized to butyrate 

and has favorable pharmacokinetics compared with butyrate, 

with low toxicity. Methods: In the present study, we used a 

mouse model of ALD to examine the effects of Tb (2g/kg) oral 

administration on ethanol-induced changes in hepatic inflammation 

and injury. 8–10-week old C57BL/6 male mice were 

pair-fed the Lieber-DeCarli liquid diet (Research Diets, NJ). containing 

alcohol (AF, n =8) or isocaloric maltose dextrin (PF, n 

=8) for 2 and 7 weeks. Tb was administered to a sub-group of 

alcohol-fed animals by oral gavage for 5 days/week. Serum 

and liver tissue samples were analyzed for hepatic inflammation 

and injury. Results: At the end of 7 weeks Tb significantly 

attenuated the ethanol-induced neutrophil infiltration as shown 

by choline esterase staining. Quantification of neutrophil infiltration 

by myeloperoxidase (MPO) activity showed similar 

results wherein Tb administered mice had a significant drop 

in MPO activity as compared to alcohol-fed mice at 7 weeks 

but not in 2 weeks. F4/80 staining of liver sections revealed 

increased macrophage infiltration in 7 weeks of alcohol feeding 

whereas Tb administration markedly blocked macrophage 

infiltration. Further, neutrophil chemoattractant chemokines 

such as CXCL-2 and CCL-2 were significantly upregulated in 

alcohol-fed mice. Importantly, Tb administration significantly 

attenuated alcohol-induced induction of these pro-inflammatory 

chemokines. Additionally, alcohol-induced TNF-α, a hallmark 

pro-inflammatory cytokine was also attenuated by Tb. Notably, 

attenuation of neutrophil infiltration and inflammation by Tb 

also led to a significant decrease in liver injury as indicated by 

serum AST & ALT levels. Conclusion: The present work demonstrates 

that Tb prevents ethanol-induced inflammation and neutrophil-mediated 

hepatic injury, and has therapeutic potential 

in the prevention/treatment of ALD. 

Disclosures: 





The following authors have nothing to disclose: Hridgandh Donde, Smita Ghare, 

Jingwen Zhang, Swati Joshi-Barve


1332 

Differential microRNA expression profiles in patients 

with alcoholic hepatitis (AH): Preliminary findings from 

the Translational Research and Evolving Alcoholic Hepatitis 

Treatment (TREAT) consortium 

Suthat Liangpunsakul 1 , Naga P. Chalasani 1 , Guanglong Jiang 2 , 

Yunlong Liu 2 , Vijay Shah 3 , Arun J. Sanyal 4 , Svetlana Radaeva 5 , 

David W. Crabb 1 ; 1 Division of Gastroenterology/Hepatology, 

Indiana University School of Medicine, Indianapolis, IN; 2 Department 

of Medical and Molecular Genetics, Center for Computational 

Biology and Bioinformatics, Indiana University, Indianapolis, 

IN; 3 Division of Gastroenterology and Hepatology, Department of 

Medicine, Mayo Clinic College of Medicine, Rochester, MN; 4 Division 

of Gastroenterology and Hepatology, Department of Medicine, 

Virginia Commonwealth University, Richmond, VA; 5 NIH/ 

NIAAA, Rockville, MD 

Background: It is poorly understood why only a subset of subjects 

who drink excessively will develop AH. microRNAs (miR- 

NAs) are major regulators of multiple biological processes. 

Alterations in the miRNAs might play a role in the pathogenesis 

of AH. Aim: To determine differentially expressed serum miR- 

NAs in subjects with AH compared to heavy drinking controls 

without liver disease. Methods: AH was diagnosed as those 

with (i) alcohol consumption of >40 g/d for women and >60 

g/d for men for a minimum of 6 mos and (ii) total bilirubin>2 

mg/dL and AST > 50 U/L. Controls were heavy drinkers with 

normal hepatic panel and no evidence of liver disease. RNA 

was isolated from the serum of 19 AH (mean age 45 yrs, 50% 

women, and 90% White) cases and 20 controls (mean age 44 

yrs, 50% women, and 70% White) and custom miRNA profiling 

was done by microarray analyses (LC Sciences, Houston, 

TX). Results: Array analysis consisted of 2,615 probes. 111 

miRNAs were > 2 fold differentially expressed between both 

groups with false discovery rate < 0.05. Table 1 describes top 

miRNAs with increased and decreased expression in AH compared 

to heavy drinking controls. Validated targets for these 

miRNAs were sought in the literature: Of the up-regulated miR- 

NAs, miR-765 targets HNF4, a transcription factor regulating 

many genes determining liver differentiation. Of the down-regulated 

miRNAs, the following targets have been reported: IL1 

beta for miR-376c; mTOR, c-Met (hepatocyte growth factor), 

junB, and IkB kinase for miR-199a; TGF beta and IRS1 for 

miR-487; Wnt/beta catenin for miR-432b; and multiple innate 

immunity pathway genes for miR-146b. Conclusion: We identified 

the serum miRNAs that differentially expressed between 

heavy drinkers and AH subjects. Several of these miRNAs have 

been reported to be involved in regulation of processes known 

to be important in the pathogenesis of AH, such as cytokine 

signaling, hepatic growth and differentiation, and innate immunity, 

suggesting the changes may provide clues to mechanisms 

of liver injury in AH, as well as serving as biomarkers. 

Table 1: Serum miRNAs differentially expressed between patients 

with AH and heavy drinking controls 

Disclosures: 








The following authors have nothing to disclose: Suthat Liangpunsakul, Guanglong 

Jiang, Yunlong Liu, Vijay Shah, Svetlana Radaeva, David W. Crabb 

1333 

Oral pentamidine (VLX103) prevents the development 

of alcoholic liver disease in mice 

Ghulam Ilyas 1 , Enpeng Zhao 1 , Kathryn Tanaka 3 , Francois Ravenelle 

2 , Mark J. Czaja 1 ; 1 Medicine, Albert Einstein College of 

Medicine, Bronx, NY; 2 Verlyx Pharma Inc, Montreal, QC, Canada; 

3 Pathology, Albert Einstein College of Medicine, Bronx, NY 

In alcoholic liver disease (ALD) abnormal lipid metabolism and 

hepatocyte injury are driven not only by the direct toxic effects 

of alcohol and its metabolites, but also by an overactive innate 

immune response generated by stimulation from intestinal lipopolysaccharide 

(LPS). Activated macrophages generate factors 

such as the proinflammatory cytokine tumor necrosis factor 

(TNF) which in the setting of alcohol exposure become cytotoxic 

to hepatocytes. Previously we have demonstrated that a 

novel form of oral pentamidine, VLX103, significantly reduces 

liver injury and improves survival from galactosamine/LPS. The 

efficacy of VLX103 in this form of toxic liver injury, together 

with the known anti-inflammatory and LPS-binding properties 

of pentamidine, suggested that VLX103 may be effective 

in the treatment of ALD. We therefore tested the hypothesis 

that VLX103 would reduce liver injury in the NIAAA mouse 

model of ALD. Methods: A modification of the NIAAA model 

was employed in C57BL/6 mice that consisted of 10 days of 

control or ethanol diet followed by administration of a single 

binge dose of maltose or ethanol, respectively. During the diet 

feeding mice received vehicle or 75 mg/kg of VLX103 by 

daily gavage. Mice were analyzed 9 h after the maltose/ 

ethanol gavage. Results: Treatment with VLX103 significantly 

decreased the degree of ethanol-induced steatosis as indicated 

by a 34% reduction in triglyceride accumulation (64.8 vs. 97.7 

mg/g liver; P


1334 

Inflammatory and apoptotic markers in alcoholic hepatitis 

patients: interim analyses 

Leila Gobejishvili 1 , Vatsalya Vatsalya 1 , Mohammad K. Mohammad 

1 , Shirish Barve 1 , Craig J. McClain 1,2 ; 1 Department of Medicine/GI, 



This study is part of a NIAAA UO1 consortium to identify novel 

biomarkers and unique drug targets for treatments for AH. 17 

male and female acute AH patients within the age group of 

21-70 yrs. and 10 healthy controls were included in this initial 

pilot investigation. Clinical data for demographic, drinking 

history and liver injury markers, MELD, Maddrey DF, and CTP 

were recorded. Among AH patients, 6 had a MELD score in a 

non-severe range (≤ 19) and 11 were in the severe range (≥ 

20). Milliplex analysis of serum analytes including cytokines, 

soluble FasL, Osteopontin (OPN) and TNF-related apoptosis-inducing 

ligand (TRAIL) was performed. A major finding of 

our study demonstrates significant inverse correlation of TRAIL 

expression with severity of AH represented by MELD and neutrophillia. 

AH patients often have hepatic polymorphonuclear 

leukocyte infiltration and neutrophilia which plays a major role 

in determining whether patients develop acute inflammation. 

Importantly, resolution of acute inflammation involves neutrophil 

apoptosis mediated by TRAIL. In view of this, association 

of the decreasing levels of TRAIL with increasing severity of AH 

suggests that this could be a significant pathogenic mechanism 

that contributes to the development and severity of AH. In comparison, 

there was significant direct correlation of sFasL levels 

with the severity of AH in our patient population. Fas/FasL 

mediated apoptosis plays a major role in hepatocyte death 

that occurs in the proinflammatory cytokine milieu of alcoholic 

hepatitis. Similar increase in circulating sFasL has been 

reported in severe AH patients (Maddrey score ≥32). Further, 

as reported previously, AH patients showed significantly higher 

serum Osteopontin levels compared to controls and positively 

correlated with MELD. Overall, the findings of our initial pilot 

study have begun to identify relevant AH-specific pathogenic 

determinants that could serve as prognostic biomarkers and 

targets for intervention. 

Significant inverse correlation between serum TRAIL levels and 

MELD in AH patients. 

1335 

Zinc deficency inactivate hepatic mitochondrial biogensis 

pathway in rats chronically fed alcohol: A mechanism 

of perturbed hepatic mitochondrial respiratory 

complexes and ROS generation 

Qian Sun, Wei Zhong, Wenliang Zhang, Zhanxiang Zhou; Nutrition, 

University of North Carolina at greensboro, Kannapolis, NC 

Clinical studies demonstrated that patients with alcoholic liver 

disease (ALD) have a decreased hepatic zinc level, however, 

the effect of zinc deficiency on the function of hepatic mitochondria 

has not been well studied. The present study was 

undertaken to determine if zinc deficiency causes mitochondrial 

electron transport chain (ETC) defect and if defected ETC 

function could increase hepatic oxidative stress in the presence 

of fatty acid and acetaldehyde. Wistar rats were pair-fed with 

the Lieber-DeCarli control or ethanol diet for 5 month. Chronic 

alcohol exposure significantly increased hepatic triacylglycerol, 

free fatty acid and 4-hydroxynonenal (4HNE) levels, meanwhile 

hepatic mitochondrial 4HNE level was also increased 

and mitochondrial zinc level was reduced. In addition, hepatic 

mitochondrial respiratory complex I, III, IV and V were significantly 

decreased by chronic alcohol exposure, which was in 

consistence with decreased hepatic ATP production. Mitochondrial 

biogenesis signal transduction pathway was impaired 

by chronic alcohol feeding as indicated by decreased AMPK, 

PGC1αa, NRF1 and TFAM levels and mitochondrial DNA 

expression. In order to define the link between zinc deficiency 

and the defect of mitochondrial respiratory complexes, hepG2 

cells were treated with TPEN for 6h. The results indicated that 

zinc deficiency significantly decreased mitochondrial respiratory 

complex I, III, IV expression. In addition, AMPK, PGC1α, 

NRF1 and TFAM levels, and mitochondrial DNA expression 

were significantly decreased by TPEN treatment. Consequently, 

mitochondrial respiratory complex I, III, IV was silenced by 

shRNA, respectively. The transfected hepG2 cells all showed 

a decrease in mitochondrial membrane potential and an 

increase in free radical production after challenged with 500 

mM linoleic acid or 200mM acetaldehyde for 6h. These results 

suggest that alcohol exposure induced hepatic zinc deficiency 

could inactivate mitochondrial biogenesis signal transduction 

pathway and decrease mitochondrial DNA production, which, 

in turn, decreases mitochondrial complex protein expression. 

The defect of mitochondrial respiratory complexes could 

worsen alcohol consumption induced free radical production 

while metabolizing fatty acid and acetaldehyde. 

Disclosures: 

The following authors have nothing to disclose: Qian Sun, Wei Zhong, Wenliang 

Zhang, Zhanxiang Zhou 

Disclosures: 





The following authors have nothing to disclose: Leila Gobejishvili, Vatsalya Vatsalya, 

Mohammad K. Mohammad 

1336 

Nuclear receptor Rev-Erbα functions as a transcriptional 

activator of Chop to control SHP mediated alcoholic 

fatty liver 

zhihong yang 1 , Hiroyuki Tsuchiya 2 , Sangmin Lee 1 , Yuxia Zhang 3 , 

Li Wang 1 ; 1 PNB, university of Connecticut, Hamden, CT; 2 School 

of Medicine, university of Utah, Salt Lake City, UT; 3 Pharmacology, 

Toxicology & Therapeutics, University of Kansas Medical Center, 

Kansas City, KS 

[Purpose] Excessive consumption of alcohol results in development 

steatosis and steatohepatitis, which can progress to 

cirrhosis and hepatocellular carcinoma. The endoplasmic reticulum 

(ER) stress contributes to metabolic disturbances and is 

associated with alcoholic liver disease (ALD). This project aims 

at identifying new molecular mechanisms in nuclear receptor


mediated circadian regulation of alcoholic fatty liver. [Methods] 

The ethanol-binge model (NIAAA model) used in the 

current study is described previously. Briefly, two months old 

C57BL/6 (WT) and nuclear receptor Shp-/- mice were fed 

with Lieber-DeCarli diet containing 5% ethanol ad libitum or 

pair- fed with the isocaloric control diet for 10 days. On day 

11, mice were oral-gavaged with a single dose of 5g of ethanol/kg 

of body weight (EtOH) or maltose dextrin (CTRL). After 

nine hours, the serum and liver samples were collected every 

six hours for twenty-four hours, subjected to mRNA analysis by 

qPCR and RNA-seq or protein expression analysis by Western 

Blotting. [Results] Liver TG levels, lipid accumulation and glycogen 

increased significantly in WT mice after ethanol binge 

compared to control group fed isocaloric diet, but decreased 

in Shp -/- mice. Surprisingly, Shp-/- mice receiving control diet 

exhibited higher basal levels of TG and lipid accumulation. 

Srebp1c mRNA and protein expression were also elevated 

in Shp -/- mice, but were reduced in EtOH group, as well as 

Fasn, one of Srebp1c’s targets. ER stress markers p-eIf2a and 

Atf4 protein increased due to alcohol treatment in WT mice 

and the increase is abrogated in Shp-/- mice. The transcription 

factor C/EBP homologous protein (Chop) is an evidenced 

downstream target of ATF4. Unexpectedly, the basal Chop 

expression was higher in Shp-/- mice receiving control diet, 

Chop was decreased following alcohol binge in both WT and 

Shp-/- mice and the expression was in a circadian fashion. 

Rev-Erbα functions as a negative-regulator of several clock proteins. 

Interestingly, its levels exhibited a similar changes as 

Chop. We further demonstrated that Rev-Erbα activated Chop 

promoter activity and induced Chop mRNA and protein expression, 

which was inhibited by co-expression of Shp. In addition, 

knockdown Rev-Erbα by shRNA reduced Srebp1c protein 

cleavage. [Conclusions] Our study identified a novel function 

of Rev-Erbα to modulate SHP mediated Chop expression and 

Srebp1c cleavage, thus providing a new regulatory mechanism 

in circadian clock control of alcoholic fatty liver. 

Disclosures: 

The following authors have nothing to disclose: zhihong yang, Hiroyuki Tsuchiya, 

Sangmin Lee, Yuxia Zhang, Li Wang 

1337 

The Alcoholic Hepatitis Histology Score Performs Poorly 

in a Cohort of Severe Alcoholic Hepatitis in the United 

States 

Gene Y. Im 1 , Aparna Goel 1 , Stephen C. Ward 2 , Yujin Hoshida 1 , 

Thomas D. Schiano 1 , Scott L. Friedman 1 , Swan N. Thung 2 ; 1 Medicine, 

Division of Liver Diseases, Icahn School of Medicine at Mount 

Sinai, New York, NY; 2 Pathology, Icahn School of Medicine, New 

York, NY 

Prediction models for alcoholic hepatitis (AH) that are commonly 

used only include clinical variables. The Alcoholic Hepatitis 

Histology Score (AHHS) is a newly derived and validated 

histologic prediction model. Our aim was to evaluate the performance 

of the AHHS in a real-world cohort of severe AH 

patients in the United States. Methods: Patients hospitalized 

at a high volume liver transplantation (LT) center for severe 

AH were prospectively identified from 1/2012 to 6/2015 for 

our AH database. Patients with severe AH on liver histology 

obtained early in their hospitalization and with a Maddrey’s 

discriminant function (DF) ≥32 were included. Liver histology 

slides were reviewed by expert hepatobiliary pathologists 

who were blinded to patient characteristics. Clinical data 

were reviewed to calculate DF, Lille, model for end-stage liver 

disease (MELD), Glasgow alcoholic hepatitis score (GAHS) 

and age, bilirubin, INR, creatinine (ABIC) scores at presentation. 

The primary outcome of analysis was 90-day mortality or 

LT. Results: Over the 3.5 year study period, 120 consecutive 

patients with severe AH were admitted or transferred to our 

liver service and prospectively evaluated. Twenty-six patients 

had available liver tissue for analysis. The median age was 44 

years (IQR 38-53) and two-thirds Caucasian with female predominance. 

Median DF, MELD, Lille, GAHS and ABIC scores 

were 78 (IQR 58-93), 33 (IQR 26-39), 0.970 (IQR 0.41- 

0.99), 7.5 (7-9) and 8.9 (7.8-9.7), respectively, indicative of 

a cohort with high risk of mortality. Standard medical care for 

advanced liver disease was provided, with less than one-quarter 

receiving glucocorticoid-based therapies and half ineligible 

due to severe illness. The cumulative mortality or LT rates at 

day 30, 60, 90 and 180 were 42%, 46%, 65% and 69%, 

respectively. The AHHS model performed poorly in predicting 

90-day mortality or LT, with an area under the curve (AUC) of 

0.57. The Lille and GAHS models performed well with AUCs 

of 0.83 and 0.71, respectively, while MELD and ABIC had 

AUCs of 0.56 and 0.55, respectively. Conclusions: The AHHS 

model performed poorly in predicting 90-day mortality or LT in 

a real-world cohort of severe AH patients in the United States. 

As a static histologic score, the AHHS may lack the precision to 

capture the dynamic clinical complexity of severe AH. 

Disclosures: 













The following authors have nothing to disclose: Gene Y. Im, Aparna Goel, Stephen 

C. Ward, Yujin Hoshida, Swan N. Thung 

1338 

Genetic Ablation of MitoNEET Alleviates Alcoholic Steatosis/Steahepatitis 

in Mice 

Alvin Jogasuria 1 , Werner J. Geldenhuys 1 , Jiayou Wang 1,2 , Xudong 

Hu 1,3 , Kwangwon Lee 1 , Prabodh Sadana 1 , Jiashin Wu 1 , 

Min You 1 ; 1 Pharmaceutical Sciences, Northeast Ohio Medical 

University (NEOMED), Rootstown, OH; 2 Department of Anatomy, 

Guangzhou University of Chinese Medicine, Guangzhou, China; 

3 Department of Biology, Shanghai University of Traditional Chinese 

Medicine, Shanghai, China 

MitoNEET (CDGSH iron-sulfur domain-containing protein 1 

or CISD1) is an outer mitochondrial membrane protein that 

donates 2Fe-2S clusters to apo-acceptor proteins, and regulates 

mitochondrial matrix iron metabolism. Clinically, aberrant 

hepatic iron metabolism is frequently occurred in patients 

with alcoholic liver disease. It is also reported that, in vitro 

cultured hepatocytes, ethanol along with fructose induces a 

large increase in the expression of mitoNEET, which primed the 

fructose and ethanol exposed hepatocytes for TNFα-induced 

necroptosis. Therefore, in the present study, using a mitoNEET 

knock out (MitoNEETKO) mouse model, we aimed to investigate 

the in vivo functional role of mitoNEET in the development 

of alcoholic steatosis/steatohepatitis and explore the underlying 

mechanisms. Alcoholic fatty liver injury was achieved 

by pair feeding wild-type (WT) and MitoNEETKO mice with 

Lieber-DeCarli ethanol-containing diets for 4-wks. Remarkably, 

we found that chronically ethanol-fed MitoNEETKO mice were 

resistant to ethanol-induced fatty liver injury demonstrating by


dramatically reduced hepatic triglycerides, decreased hepatic 

cholesterol amount and normalized serum liver enzymes compared 

to ethanol-fed WT mice. Our studies revealed that genetically 

ablation of mitoNEET in mice alleviated alcoholic fatty 

liver injury by way of turning on multiples signaling pathways, 

including stimulated ileal fibroblast growth factor 15 (FGF15) 

synthesis, activated a pre-mRNA splicing regulator SLU7, and 

suppressed hepatic lipin-1-medaited phosphatidate phosphatase 

(PAP) activity. Altogether, our novel findings suggest that 

mitoNEET plays a pivotal role in development and progression 

of alcoholic fatty liver injury in mice. Hence, Pharmacological 

or nutritional modulation of mitoNEET may be beneficial for 

the prevention or treatment of human alcoholic steatosis/steatohepatitis. 

Disclosures: 

The following authors have nothing to disclose: Alvin Jogasuria, Werner J. 

Geldenhuys, Jiayou Wang, Xudong Hu, Kwangwon Lee, Prabodh Sadana, 

Jiashin Wu, Min You 

1339 

Cross talk between hepatocytes and macrophages in 

Alcoholic Liver Disease: which language do they use? 

Veronica Marin 1 , Kyle L. Poulsen 2 , Laura E. Nagy 2,3 , Claudio 

Tiribelli 1,4 , Natalia Rosso 1 ; 1 Fondazione Italiana Fegato, Trieste, 

Italy; 2 Department of Pathobiology, Cleveland Clinic, Cleveland, 

OH; 3 Department of Molecular Medicine, Case Western Reserve 

University, Cleveland, OH; 4 Department of Medical Sciences, University 

of Trieste, Trieste, Italy 

Macrophage migration inhibitory factor (MIF) is a key cytokine 

involved several inflammatory diseases that, depending on 

the target cell and inammatory context, can engage different 

receptors. It has been reported that interaction between MIF 

and CD74 could exert hepatoprotective effects. In the present 

study, we explored specifically MIF and CD74 expression in 

hepatocytes and macrophage in response to ethanol (EtOH) 

exposure. Co-culture of hepatocytes (HuH7) and differentiated 

macrophages (THP1+100nM PMA) were exposed to 25mM 

EtOH for 24h; monocultures of each cell type were used as 

controls (CTRL). Gene expression of MIF, CD74 and TNF-α 

was analyzed by real time PCR. The amount of MIF and TNF-α 

released in the cultured-media was quantified by ELISA. CD74 

protein expression was detected with anti-human CD74-FITC 

antibody by flow cytometry. In hepatocytes monoculture, EtOH 

induces an up-regulation of TNF-α and MIF mRNA expression, 

not translated in cytokines’ release. Interestingly, CD74 expression 

(gene and protein) was barely detected in these cells. 

On the other hand, in macrophages monoculture EtOH did 

not induce relevant changes in any of the parameters under 

study. When co-cultured, EtOH increased MIF release (with 

no changes at mRNA level).both in hepatocytes and macrophages. 

Interestingly, co-cultured hepatocytes showed a significant 

up-regulation of CD74 mRNA expression compared with 

monoculture CTRL and the same trend was also confirmed at 

CD74 protein level in both co-cultured cell types. Regarding 

TNF-α even if mRNA was dramatically upregulated its release 

was unchanged in hepatocytes and significantly decreased in 

macrophages. In conclusion, these data clearly demonstrate 

that there is a differential cellular response to EtOH when 

hepatocytes and macrophages are cultured together. We 

hypothesize that EtOH-challenged hepatocytes (in presence 

of macrophages) might be the main source of MIF production 

and that CD74 play a determinant role in this mechanism. The 

reduction in macrophages TNF-α release lead to the speculation 

that in this model MIF through CD74 interaction might 

exert hepatoprotective effects. 

Table 1- Summary of the obtained data both in monoculture and 

co-culture system in response to ethanol exposure 

a p


careful cut-offs from HC, was lower than in literature, adipopenia 

was more pronounced. 

Disclosures: 

The following authors have nothing to disclose: Onan Perez-Hernandez, Geraldine 

del Carmen Quintero-Platt, Carlos Jorge-Ripper, Camino Maria Fernandez-Rodriguez, 

Maria Blanca Monereo Muñoz, Maria Jose Sanchez-Perez, 

Emilio Gonzalez-Reimers, Ruben Hernandez-Luis 

Data expressed as mean± SD/ median (IQR) # (C vs. DC), *(HC 

vs.C), $ (HC vs. DC) 

Disclosures: 


inc 

The following authors have nothing to disclose: Varsha Shasthry, Jaya Benjamin, 

Chethan Kalal, Lovkesh Anand, Ankit Bhardwaj, Vanshja Pandit, Ankur Arora, S. 

Rajesh, Viniyendra Pamecha, Vikas Jain, Guresh Kumar, Y.K Joshi, Shiv K. Sarin 

1341 

The role of proinflammatory cytokines, adipokines and 

lipid peroxidation in short term mortality in patients 

with Severe Alcoholic Hepatitis 

Onan Perez-Hernandez 1,2 , Geraldine del Carmen Quintero-Platt 1 , 

Carlos Jorge-Ripper 1,2 , Camino Maria Fernandez-Rodriguez 1,2 , 

Maria Blanca Monereo Muñoz 1 , Maria Jose Sanchez-Perez 1 , 

Emilio Gonzalez-Reimers 1,2 , Ruben Hernandez-Luis 1,2 ; 1 Medicina 

Interna, Hospital Universitario de Canarias, San Cristóbal de La 

Laguna, Spain; 2 Medicina Interna, Universidad de La Laguna, San 

Cristobal de La Laguna, Spain 

INTRODUCTION Acute alcoholic hepatitis is a serious disease 

associated with a high short term mortality ranging from 20 to 

50%. The factors involved in mortality in patients with acute 

alcoholic hepatitis are still controversial. It is well known that 

acute alcoholic hepatitis is a disease in which cytokine activation 

and oxidative damage play important roles. The aim of 

this study is to prospectively assess the relationship between 

cytokines, lipid peroxidation, and adipokines with mortality 

of patients with alcoholic hepatitis at 30, 90, and 180 days. 

PATIENTS AND METHODS Sixty patients with severe alcoholic 

hepatitis (Maddrey index ≥32) were successively included. 

Cytokine levels (IL-4, IL-6, IL-8, TNF-α, INF-γ, adiponectin, 

resistin, leptin, insulin and MDA) were measured within the first 

48 hours after admission and one week later. Patients were followed-up 

during 180 days; 12 patients died within the first 30 

days, 21 within 90 days, and 23 within 180 days. RESULTS 

The mean age was 51.9 years (SD=±10.7). Serum levels at 

admission of TNF-α were significantly higher among those 

who died at admission (p=0.041). IL-4 were significantly lower 

among those who died within 30 days (p=0.048) whereas 

levels of IL-8 were associated with an increased mortality at 

90 and 180 days (p=0.022 and 0.027, respectively). Serum 

MDA levels at admission were significantly associated with 

mortality at admission, 30, 90, and 180 days. Serum leptin 

levels were also associated with mortality at 180 days and 

insulin levels were related to mortality at admission, 30, 90, 

and 180 days. CONCLUSION Twenty percent of patients died 

within the first 30 days after admission; 35.5% died within 90 

days, and 38.7% died within 180 days. Increased proinflammatory 

cytokine levels at admission and especially increased 

MDA levels are significantly associated with short-term mortality 

among alcoholics. Lower IL-4 levels were also associated 

with mortality. No relation was observed between mortality 

and adiponectin levels but indeed between mortality and leptin 

levels. Higher mortality was observed among those with low 

insulin levels. 

1342 

Nosocomial Infection in Severe Alcoholic Hepatitis 

Onan Perez-Hernandez 1,2 , Geraldine del Carmen Quintero-Platt 1 , 

Emilio Gonzalez-Reimers 1,2 , Maria Jose Sanchez-Perez 1 , Pedro 

Abreu-Gonzalez 3 , Maria Jose de la Vega-Prieto 4 , Carlos Jorge-Ripper 

1,2 , Francisco Santolaria-Fernandez 1,2 ; 1 Medicina Interna, 

Hospital Universitario de Canarias, San Cristóbal de La Laguna, 

Spain; 2 Medicina Interna, Universidad de la Laguna, San Cristobal 

de La Laguna, Spain; 3 Fisiologia, Universidad de La Laguna, 

San Cristobal de La Laguna, Spain; 4 Laboratorio Central, Hospital 

Universitario de Canarias, San Cristobal de La Laguna, Spain 

INTRODUCTION Both alcoholism and liver cirrhosis increases 

the risk of infection. In acute alcoholic hepatitis, especially 

in severe cases (Maddrey ≥32), intestinal permeability is 

increased. This fact triggers an greater risk of bacterial translocation 

and, on the one hand, more infections and on the other, 

increased lipid peroxidation mediated proinflammatory activation. 

OBJECTIVE The aim of this study is to prospectively assess 

the nosocomial infections in severe acute alcoholic hepatitis 

(sAAH) and its prognostic value. Furthermore, we examine 

the relationship between inflammatory cytokines, adipokines 

and lipid peroxidation with nosocomial infections. MATERIAL 

AND METHOD Sixty-two patients with sAAH were successively 

included. Nosocomial infections were registered. Cytokine levels 

(IL-4, IL-6, IL-8, TNF-α, INF-γ, adiponectin, resistin, leptin, 

insulin and MDA) were measured within the first 48 hours 

after admission and one week later. Patients were followed-up 

during 180 days. RESULTS The mean age was 52.2 years 

(±10.7). Maddrey’s index on admission was 55.6 (±19.8). 

29.5% of patients had a infection during hospitalization 

(66.7% were pneumonia), which caused 42.9% of deaths. A 

higher percentage of patients with nosocomial infection died 

at 180 days (58.8 vs 28.9%; p = 0.03). Infection appeared 

after 22.8 days (±16.7). There was a trend to hyponatremia 

at admission was more frequent (68.8 vs 31.3%; p = 0.064). 

After the first week, infected patients had a worsening of bilirubin 

(76.5 vs 48.8%; p=0.048), ABIC≥8 (73.3 vs 40.5%; 

p=0,032), platelets 7.91; p=0.031). CONCLUSION Nosocomial 

infections were a common complication in patients admitted for 

sAAH and were the main cause of death. Data exist relating of 

develop it as unfavorable evolution in the first week, measured 

by ABIC index, and worsening of bilirubin levels. Thrombocytopenia 

and hypofibrinogenemia also associated with increased 

risk. Infected patients had lower adiponectin levels, both at 

admission and first week. Finally, we also observed lipid peroxidation 

was higher in this group. 

Disclosures: 

The following authors have nothing to disclose: Onan Perez-Hernandez, Geraldine 

del Carmen Quintero-Platt, Emilio Gonzalez-Reimers, Maria Jose Sanchez-Perez, 

Pedro Abreu-Gonzalez, Maria Jose de la Vega-Prieto, Carlos 

Jorge-Ripper, Francisco Santolaria-Fernandez


1343 

Effects of Alcohol and Abstinence on Liver Stiffness as 

measured by Transient Elastography 

Izabella Pokorski 1 , Yang Wu 2 , Martin Weltman 2 , Guy D. Eslick 3 ; 

1 Centre for Addiction Medicine, Nepean Blue Mountains Local 

Health District, Penrith, NSW, Australia; 2 Gastroenterology and 

Hepatology, Nepean Hospital, Penrith, NSW, Australia; 3 Surgery, 

The University of Sydney, Penrith, NSW, Australia 

The purpose of this study is to examine the relationship between 

alcohol consumption, monitored abstinence, relapse and liver 

stiffness as measured by Transient Elastogprahy (TE) to elucidate 

the best time to accurately record liver fibrosis in patients 

with Alcoholic Liver Disease (ALD). TE is a potential alternative 

to biopsy to measure liver stiffness, an indirect estimation of 

liver fibrosis. Patients presenting for voluntary detoxification 

of their alcohol dependence at Nepean Hospital were consecutively 

approached from June 2014 until May 2015. Liver 

stiffness measurements were recorded on admission, discharge 

and at least 7 days post discharge. Relapse information after 

discharge was recorded. Blood tests as well as BMI, hepatitis 

B and C status were collated for sub-analysis. 123 patients 

were recruited.13% were excluded due to their body habitus 

and early discharge limited second scan data. 76% of participants 

were male (53/70) and 24% of participants were female 

(17/70). 70 patients had 2 valid scans, with an average mean 

reduction of -2.25kPa in liver stiffness from day of admission 

to second TE scan (statistically significant P>0.001). 47% of 

patients showed a decrease in at least 1 fibrosis stage (statistically 

significant P < 0.001).The average length between admission 

and discharge scan was 4 days (range 1-31). Patients 

were put into 2 groups based on days between TE scans (3 

days and under, more than 3 days). The difference in TE score 

for patients who were scanned after 3 days and above was 

more statistically significant (P < 0.001) compared to patients 

scanned in 3 days and under (P =0.004). 19 (27%) patients 

attended a third follow-up scan with a mean time of 96 days 

from discharge. Difference in measurements was not statistically 

significant. Fibrosis severity among patients with ALD not 

infected with HCV (n=40) was variable (F0-1 = 55%, F2-F3 

= 35%, F4 = 10%). 19% of patients reported having been 

diagnosed with HCV with 46% of these patients initially found 

to have cirrhosis (14.8kPa – 66.4 kPa). The study suggests that 

liver stiffness as measured by TE declines in patients with ALD, 

Hepatitis C and Cirrhosis while undergoing monitored abstinence 

from alcohol. A decrease in stage of fibrosis is clinically 

significant. The small number of patients presenting for a third 

TE scan after discharge (n=19) has made it difficult to evaluate 

the effect of relapse of alcohol consumption. However, it is 

clear that ongoing consumption of alcohol facilitates over-reading 

of TE scores. To obtain the most accurate TE reading, a 

scan should be performed at least 3-5 days post complete 

alcohol abstinence. 

Disclosures: 

The following authors have nothing to disclose: Izabella Pokorski, Yang Wu, 

Martin Weltman, Guy D. Eslick 

1344 

Microparticles Are Markers Of Immune Cell Stress And 

Predict Severity And Outcomes In Patients With Alcoholic 

Liver Disease 

Sukriti Sukriti 1 , Jaswinder S. Maras 1 , Shvetank Sharma 1 , Madhumita 

Premkumar 2 , Sukanta Das 1 , Shabir Hussain 1 , Guresh Kumar 1 , 

Naminita Gogoi 3 , Ashok K. Choudhury 2 , Chinmay Mukhopadhyay 

3 , Nirupma Trehanpati 1 , Shiv K. Sarin 1,2 ; 1 Research, Institute 

of Liver and Biliary Sciences, New Delhi, India; 2 Hepatology, 

Institute of Liver and Biliary Sciences, New Delhi, India; 3 Special 

Center for Molecular Medicine, JNU, New Delhi, India 

Background and Aims: Microparticles (MPs) are the membrane 

bound vesicles released during cellular stress. Disease 

specific MP signatures may aid disease progression or treatment 

outcomes. Severe alcoholic hepatitis (SAH) has a rapid 

progression and poor response to current therapies. Reliable 

tools, preferably noninvasive, are needed for early assessment 

of progression and treatment response. We investigated the 

correlation between MPs released in SAH, steroid responders 

(R) and non-responders (NR). Patients and Methods: MPs 

were isolated from plasma using differential ultracentrifugation 

followed by flow cytometry. MPs were enumerated by addition 

of known size beads (0.22,0.44,0.88,1.45um). This size 

gate was combined with Annexin V staining and, various cell 

specific markers were used. Reference counting beads were 

added to determine absolute count of MPs/ml. MPs were determined 

at day 0 and 7 in all 40 patients with SAH; untreated 

(n=8), steroid responder (R)(n=20), NR (n=12), and healthy 

controls (n=20). The MP counts were correlated with MELD 

score, Lille score, serum bilirubin and ALT/AST. Results: MPs 

associated with HSCs (CD34+), macs (CD68+) and Tcells 

(CD3+CD8+) were higher in NR at day0 (p


1345 

An alarmin cytokine IL-33 inhibits type I NKT cells and 

neutrophil accumulation into liver and mediates attenuation 

of alcoholic liver disease 

Igor Maricic, Ekihiro Seki, Idania Marrero, Vipin Kumar; Medicine, 

University of California, La Jolla, CA 

Natural killer T (NKT) cells, comprised of at least two distinct 

subsets, type I and type II, recognize different lipid antigens 

presented by CD1d molecules and are involved in inflammatory 

liver diseases. We have recently found that type I but not 

type II NKT cells become activated and mediate liver injury following 

chronic plus binge feeding of Lieber-DeCarli liquid diet 

in male C57BL/6 mice. Here we have analyzed the hepatic 

cytokine gene expression using quantitative PCR, and found 

that IL-33 is significantly upregulated during alcoholic liver disease 

(ALD). Interestingly elevated levels of IL-33 are dependent 

upon the presence of type I NKT cells as no upregulation was 

observed in Jα18-/- mice. Additionally, inhibition of type I NKT 

cells by sulfatide or retinoids also blunts IL-33 levels. Next we 

examined the effect of IL-33 treatment on ALD. Surprisingly i.p. 

administration of IL-33 (2 μg/mouse) resulted in the amelioration 

of liver injury whereas anti-IL-33 (5 μg/mouse) treatment 

leads to exacerbation of liver enzymes and hepatosteatosis 

following alcohol ingestion. Notably IL-33 inhibits the proliferation 

and effector function of type I NKT cells in both in vitro 

and in vivo assays. Immunohistological analysis of liver sections 

showed that the IL-33 administration leads to a significant 

inhibition of Ly6G+ neutrophils accumulation into liver. 

In contrast anti-IL-33 treatment results in a significant decrease 

in neutrophils and an increase in F4/80+ macrophage populations. 

These studies indicate that in this model of ALD, innate 

cytokines such as IL-33 secreted by liver cells have a protective 

role from injury and is regulated by interactions with type I NKT 

cells and myeloid cells. Collectively these results suggest that 

complex interactions of different innate cells play a central role 

in mediating inflammatory liver disease and may offer novel 

therapeutic targets. 

Disclosures: 

Ekihiro Seki - Consulting: Merck, Tobira; Grant/Research Support: Nippon Zoki 

Vipin Kumar - Management Position: GRI bio 

The following authors have nothing to disclose: Igor Maricic, Idania Marrero 

1346 

Macrophage Migration Inhibitory Factor is Protective in 

a Model of Chronic-Binge Ethanol Feeding in Mice 

Kyle L. Poulsen 3 , Natalia Rosso 1 , Veronica Marin 1 , Megan 

R. McMullen 3 , Adam R. Morris 3 , Claudio Tiribelli 1,2 , Laura E. 

Nagy 3,4 ; 1 Fondazione Italiana Fegato-Centro Studi Fegato, Italian 

Liver Foundation - Liver Research Center, Trieste, Italy; 2 Clinica 

Patologie del Fegato, Universita degli Studi di Trieste, Trieste, Italy; 

3 Pathobiology, Center for Liver Disease Research - Cleveland Clinic 

Foundation, Cleveland, OH; 4 Gastroenterology, Center for Liver 

Disease Research - Cleveland Clinic Foundation, Cleveland, OH 

Chronic alcohol abuse is a leading cause of preventable morbidity 

and mortality worldwide. Ongoing and continued alcohol 

intake by alcohol abusers is the most significant risk factor 

associated with the development and progression of Alcoholic 

Liver Disease (ALD), which encompasses a spectrum of liver-associated 

pathologies. Macrophage Migration Inhibitory Factor 

(MIF), a regulator of innate immunity with chemokine- and 

cytokine-like activities, was previously identified as a key contributor 

to the early stage of ALD after chronic ethanol feeding 

to mice. While chronic ethanol feeding in mice results in mild 

liver injury and steatosis, the recently developed Chronic-Binge 

(Gao-binge) ethanol feeding protocol may better model of the 

more severe condition of acute alcoholic hepatitis in humans. 

The goal of the current study was to investigate the role of MIF 

in liver injury in response to Chronic-Binge ethanol exposure 

utilizing female C57Bl/6J and MIF -/- mice. In contrast to the 

protection of MIF-/- mice from chronic ethanol feeding, MIF - 

/- 

mice were not protected from ethanol-mediated liver injury 

in response to chronic-binge exposure, exhibiting exacerbated 

liver injury as indicated by plasma ALT and AST activities. 

Expression of pro-inflammatory cytokines TNFα and IL-1β, as 

well as chemokines MCP-1 and MIP2, were enhanced in livers 

of MIF -/- mice, further implicating MIF-mediated protection 

following Chronic-Binge ethanol feeding. Adhesion molecules 

CD62E and ICAM-1 were also enhanced in MIF -/- mice suggesting 

that MIF limits leukocyte infiltration into the liver parenchyma. 

Flow-cytometric analysis of liver non-parenchymal cells 

confirmed that Chronic-Binge ethanol feeding increased infiltration 

of CD45 + CD11B + Ly6C + (monocyte/ macrophage) and 

CD45 + CD11B + Ly6G + (neutrophil) cells into the liver. While 

MIF did not affect recruitment of Ly6C + cells, Ly6G + cells were 

increased in MIF -/- mice, consistent with a role for neutrophils 

in mediating inflammation in response to chronic binge ethanol 

exposure. Taken together, these results suggest that MIF 

protects from ethanol-mediated liver damage by limiting pro-inflammatory 

cytokine/chemokine synthesis, adhesion marker 

expression and neutrophil infiltration following Chronic-Binge 

ethanol feeding. 

Disclosures: 

The following authors have nothing to disclose: Kyle L. Poulsen, Natalia Rosso, 

Veronica Marin, Megan R. McMullen, Adam R. Morris, Claudio Tiribelli, Laura 

E. Nagy 

1347 

Creatine Supplementation Does Not Prevent Alcoholic 

Steatosis 

Dan Feng 1,2 , Ryan W. Barton 2 , Paul G. Thomes 3 , Dean J. Tuma 1,2 , 

Natalia A. Osna 1,2 , Kusum K. Kharbanda 1,2 ; 1 Research Service, 

Veterans Affairs Nebraska-Western Iowa Health Care System, 

Omaha, NE; 2 Department of Internal Medicine, university 

Nebraska Medical Center, Omaha, NE; 3 Liver Pathobiology Laboratory, 

Cannon Research Center, Carolinas Healthcare System, 

Charlotte, NC 

In our ongoing investigation in understanding the pathogenesis 

of alcohol induced injury, we have reported that chronic 

ethanol intake lowers the hepatocellular S-adenosylmethionine 

(SAM) to S-adenosylhomocysteine (SAH) ratio. The reduced 

ratio significantly impairs the activities of several SAM-dependent 

methyltransferases, leading to the generation of many 

hallmark features of alcoholic liver injury (Kharbanda, Sem 

Liv Dis, 2009). One such methyltransferase, guanidinoacetate 

methyltransferase (GAMT), is severely impaired after ethanol 

exposure that results in reduced hepatic creatine production. 

This causes detrimental consequences in the liver but also affect 

distal organs such as the heart, muscle and brain that depend 

on a steady supply of creatine from the liver. Since creatine 

supplementation has been recently reported to prevent high-fat 

diet-induced hepatic steatosis (Deminice, J Nutr, 2011), we 

sought to examine whether creatine supplementation could prevent 

alcoholic liver injury. Further, since GAMT is a major consumer 

of SAM, we were also interested in examining whether 

creatine supplementation could spare this metabolite to preserve 

hepatocellular SAM:SAH ratio as well as restore the 

depleted hepatic and extrahepatic creatine levels. Adult male 

Wistar rats were pair-fed the Lieber DeCarli control or ethanol 

diet in the presence or absence of 1% creatine in these respec-


tive diets for 4-5 weeks. At the end of the feeding regimen, the 

rats were sacrificed and blood, livers and relevant extrahepatic 

tissues were collected and processed for biochemical and histological 

analyses. We observed micro- and macro-vesicular 

steatosis and a 4 fold-increased triglyceride accumulation in 

the livers of rats fed the ethanol diet compared to the controls. 

Creatine supplementation neither prevented alcoholic 

steatosis nor decreased elevated plasma ALT levels. The lower 

hepatocellular SAM:SAH ratio seen in the ethanol-fed rats was 

also not normalized when these rats were fed the creatine 

supplemented ethanol diet nor were SAM levels increased in 

these rats. However, a >10-fold increased level of creatine 

was observed in the liver, serum, muscles and hearts of rats fed 

the creatine-supplemented control and ethanol diet. Overall, 

dietary creatine supplementation did not prevent alcoholic liver 

injury despite its known efficacy in preventing high-fat diet-induced 

steatosis. To conclude, creatine is ineffective in protecting 

against the development of alcoholic steatosis. Betaine, a 

pro-methylating agent that maintains hepatocellular SAM:SAH 

and prevents alcoholic steatosis still remains our best option for 

this treatment. 

Disclosures: 

The following authors have nothing to disclose: Dan Feng, Ryan W. Barton, Paul 

G. Thomes, Dean J. Tuma, Natalia A. Osna, Kusum K. Kharbanda 

1348 

The L-carnitine alleviate hepatic fibrosis in a non-alcoholic 


Hajime Sunagozaka 1 , Masao Honda 1 , Taro Yamashita 1 , Hikari 

Okada 1 , Naoki Oishi 1 , Tetsuro Shimakami 1 , Kazuya Kitamura 1 , 

Kuniaki Arai 1 , Yoshio Sakai 1 , Tatsuya Yamashita 1 , Naoto 

Nagata 2 , Toshinari Takamura 2 , Eishiro Mizukoshi 1 , Shuichi 

Kaneko 1 ; 1 Department of gastroenterology, kanazawa university 

Hospital, Kanazawa, Japan; 2 Department of Disease Control and 

Homeostasis, Kanazawa University Graduate School of Medical 

Sciences, kanazawa Ishikawa, Japan 

BACKGROUND & AIMS Non-alcoholic steatohepatitis (NASH) 

is a known metabolic disorder of the liver. No treatment has 

been conclusively shown conclusively to improve NASH or 

prevent disease progression. The function of L-carnitine to 

modulates the lipid profile, oxidative stress, and inflammatory 

responses. In this study, we evaluated the efficacy of L-carnitine 

for the prevention of NASH. METHODS Eight-week-old male 

C57BL/6J mice were divided into four groups: (1) basal diet, 

(2) atherogenic high-fat (Ath+HF) diet, (3) Ath+HF+0.5% carnitine 

diet, and (4) Ath+HF+1% carnitine diet. Liver histology 

and gene expression profiles were evaluated at base line and 

after 12, 30weeks of L-carnitine administration. Furthermore, 

in the clinical research, we administered 1800 mg L-carnitine 

for 24 weeks to the patients who have beenwere diagnosed 

histologically with histologically NASH for 24 weeks. 

We evaluated liver enzymes, lipid and glucose profiles, and 

histological scores at the start and end of treatment. Gene and 

protein expression were was evaluated by qRT-PCR and western 

blotting, respectively. Metabolome analysis was performed 

with mice mouse and Hhuman liver tissue samples to detect the 

changes of in metabolic path ways with induced by L-carnitine 

administration. RESULTS the L-carnitine significantly decreased 

liver weight and triglyceride TG and total cholesterol TC levels 

compared with the Ath+HF groups in a dose- dependent 

manner. Furthermore, the L-carnitine, as compared with the 

Ath+ & HF group, was associated with significant reductions 

in hepatic steatosis, lobular inflammation, and fibrosis score 

at 12 and 30 weeks in a dose- dependent manner. qRT-PCR 

analysis demonstrated that L-carnitine significantly reduced 

the alteredation of expression of the pro-fibrotic, inflammatory, 

and fat synthesis-associated genes, whereas the expression 

of PPAR-alpha expression was significantly increased. In 

Human researchthe NASH patients, L-carnitine -treated patients 

showedadministration significantly improvedments in the liver 

enzymes, and histological scores at the end of the treatment 

period. In metabolome analysis, the pentose phosphate pathway 

was accelerated with L-carnitine, whereas the glycolysis 

pathway was not activated directlyory. In this pathway, a 

large amount of NADPH was produced and the glutathione 

redox ratio was increased in the liver tissue. CONCLUSIONS 

L-carnitine supplementation improved hepatic steatosis, inflammation, 

fibrosis. L-carnitine could reduce an oxidativeon stress 

through the modification of the pentose phosphate pathway 

in the NASH liver. L-carnitine could be serve as a candidate 

therapeutic strategy for NASH. 

Disclosures: 







The following authors have nothing to disclose: Hajime Sunagozaka, Masao 

Honda, Taro Yamashita, Naoki Oishi, Tetsuro Shimakami, Kazuya Kitamura, 

Kuniaki Arai, Yoshio Sakai, Tatsuya Yamashita, Naoto Nagata, Toshinari 

Takamura, Eishiro Mizukoshi 

1349 

Functional role of the Lin28/let-7 system during alcoholic 

liver injury 

Kelly McDaniel 2,1 , Tami Annable 2,1 , Yuyan Han 3,2 , Tianhao 

Zhou 3,2 , Julie Venter 3,2 , Ying Wan 1,2 , Jessica S. Garner 1 , Nan 

Wu 3,2 , Shannon S. Glaser 3,2 , Heather L. Francis 1,2 , Gianfranco 

Alpini 3,2 , Fanyin Meng 1,2 ; 1 Scott & White Hospital, Texas A&M 

HSC College of Medicine, Temple, TX; 2 Central Texas Veterans 

Healthcare System, Temple, TX; 3 Texas A&M HSC College of Medicine, 

Temple, TX 

Background: Alcoholic liver disease (ALD) is a serious health 

concern affecting millions of patients each year. Progression of 

alcoholic liver disease (ALD) involves steatosis, inflammation 

and subsequent fibrosis leading to cirrhosis of the liver. microR- 

NAs including let-7 family have the therapeutic potentials to 

recover the liver injury and fibrosis. Lin28 has been shown 

to bind to the let-7 pre-microRNA and subsequently block the 

maturation of let-7. Our aim is to evaluate the role of lin28/ 

let-7 system in alcoholic-induced fibrotic liver disease. Methods: 

Lin28a conditional mutant mice (Lin28a tm1.2Gqda/J ) and WT 

controls underwent chronic and binge ethanol feeding (NIAAA 

model). RNA was extracted from whole liver and analyzed 

via PCR array and qPCR for fibrosis markers and microRNA. 

PCR array data was further analyzed with the GO Enrichment 

Analysis tool. Serum from mice was analyzed via ELISA for 

TGF-beta levels. Immunohistochemistry (IHC) was performed 

on liver sections for fibrosis markers and morphological analysis. 

Results: The total liver histopathology score significantly 

increased in ethanol WT group relative to control WT mice, 

and decreased in Lin28 mutant group relative to WT control 

mice with ethanol treatment. By PCR array, ethanol treated 

Lin28a mutant mice showed increase in genes associated with 

low-density lipoprotein particle receptor, inflammation, vitamin 

D synthesis and calidiol 1-monooxygenase activity compared 

to Lin28a mutant mice. Lin28a mutant mice treated with ethanol 

also showed a decrease in genes associated with primary 

miRNA processing and foregut morphogenesis compared to 

Lin28a mutant mice. Compared to wild type animals treated 

with ethanol, Lin28a mutant mice treated with ethanol showed


decreased levels of genes associated with wound healing, 

MMP regulation and angiogenesis. Quantitative PCR demonstrated 

a significant decrease in alpha-SMA and TIMP3 and a 

significant increase in let-7a in Lin28a mutant ethanol treated 

mice compared to Lin28a mutant controls. Furthermore, H&E 

staining of liver sections showed increased vascularization 

and steatosis in all animals treated with ethanol compared 

to untreated animals. Significantly reduced Sirius red staining 

was observed in ethanol treated Lin28a mutant mice relative 

to WT ethanol controls, and in Lin28a mutant mice compared 

to wild type animals. Conclusions: Lin28a/let-7 axis regulates 

innate inflammation and fibrotic prone processes and diminishes 

alcoholic liver injury. Modulation of Lin28a/let-7 system 

could be the potential therapeutic approach for alcoholic liver 

disease. 

Disclosures: 

The following authors have nothing to disclose: Kelly McDaniel, Tami Annable, 

Yuyan Han, Tianhao Zhou, Julie Venter, Ying Wan, Jessica S. Garner, Nan Wu, 

Shannon S. Glaser, Heather L. Francis, Gianfranco Alpini, Fanyin Meng 

1350 

Elevated leukocyte count combined with radiologic evidence 

of a nodular liver surface in uninfected patients 

strongly predicts histologic alcoholic hepatitis, obviating 

the need for liver biopsy 

Nitzan Roth 1 , Behnam Saberi 2 , Jared Macklin 3 , John Donovan 1 , 

Andrew Stolz 1 , Gary C. Kanel 4 , Neil Kaplowitz 1 ; 1 Division of GI/ 

Liver Diseases, University of Southern California, Los Angeles, CA; 

2 Division of Gastroenterology and Hepatology, The Johns Hopkins 

University, Baltimore, MD; 3 Department of Medicine, University of 

Southern California, Los Angeles, CA; 4 Department of Pathology, 

University of Southern California, Los Angeles, CA 

Background: The clinical presentation of alcoholic hepatitis 

(AH) can be mimicked by other alcoholic liver diseases. Our 

aim was to identify the clinical features that predict having histologic 

evidence of AH in order to define a profile that would 

obviate the need for liver biopsy. Methods: An expert hepatopathologist 

blinded to clinical data retrospectively reviewed 

the liver biopsies of 95 active alcoholic patients hospitalized 

for presumed severe AH with a discriminant function of >=32 

and a bilirubin level of >=5 mg/dL. Patients were defined as 

having definite histologic evidence of AH if lobular inflammation 

and hepatocyte damage as indicated by moderate to 

severe ballooning or easily seen Mallory-Denk bodies were 

found. Patients were defined as having no AH if there was 

no ballooning or Mallory-Denk bodies and absent or minimal 

lobular inflammation. Patients were defined as having possible 

AH if they did not meet the histologic criteria for either definite 

AH or no AH. Results: Fifty patients (53%) had definite AH and 

33 patients (35%) had possible AH. Of the 12 patients (12%) 

without AH, five had cirrhosis; the other seven with non-cirrhotic 

liver disease had alcoholic foamy degeneration or alcoholic 

fatty liver with cholestasis. Thirty-day survival was 94% 

for definite AH, 91% for possible AH, and 100% for those with 

no AH. Liver surface nodularity on imaging was 98% sensitive 

and 38% specific for histologic cirrhosis. In an analysis limited 

to uninfected patients with definite AH or no AH, greater leukocyte 

count at admission and liver surface nodularity were 

independent predictors of definite AH on biopsy (P=14x10 9 /L had 

a sensitivity of 43% and a specificity of 100% for diagnosing 

definite AH. If a nodular liver surface was present, a leukocyte 

count of >=10x10 9 /L had a sensitivity of 80% and a specificity 

of 100% for diagnosing definite AH. Using these cut offs, 

44% of our cohort met clinical criteria for AH, including 12 

of the 33 patients (36%) initially classified as possible AH on 

biopsy. Conclusions: The combination of an elevated leukocyte 

count and a nodular liver surface in the absence of infection 

retrospectively identified patients with a high likelihood of having 

histologic AH and suggests that a liver biopsy may not be 

necessary in this subset. For patients with suspected severe AH 

who do not fulfill these criteria, liver biopsy remains important, 

as 23% of these patients have no histologic AH. 

Disclosures: 

The following authors have nothing to disclose: Nitzan Roth, Behnam Saberi, 

Jared Macklin, John Donovan, Andrew Stolz, Gary C. Kanel, Neil Kaplowitz 

1351 

Molecular chaperone Hsp90 regulates cell-specific 

induction of epigenetic genes important in macrophage 

activation and lipid metabolism during alcoholic liver 

injury 

Pranoti Mandrekar, Aditya Ambade, Arlene Lim; Medicine, University 

of Massachusetts Medical School, Worcester, MA 

Background and Aims: Acetylation and methylation of chromatin-modifying 

enzymes and epigenetic genes by alcohol 

influence transcriptional activity of genes and impact alcoholic 

liver disease (ALD). Cellular stress induced molecular chaperones 

particularly hsp90 are being recognized as mediators of 

environmental impacts on epigenetic states and transgenerational 

inheritance. We hypothesized that chronic alcohol exposure 

modulates expression of chromatin modifying enzymes 

chaperoned by hsp90 during liver injury. Methods: To test 

our hypothesis, C57BL/6 mice were subjected to the NIAAA- 

Gao chronic-binge alcohol feeding model using Lieber-deCarli 

diet with 5% v/v ethanol. A single injection of hsp90 inhibitor, 

17-DMAG [17-Dimethylamino-ethylamino-17-demethoxygeldanamycin] 

was administered (30-50 mg/kg BW) i.p. 

before the binge. Serum ALT, liver cytokine expression and 

steatosis were assessed. Hsp90α was estimated in liver nuclear 

fractions. Chromatin modifying enzyme expression was analyzed 

in whole livers, isolated hepatocytes and Kupffer cells 

using PCR arrays. Results: Gene ontology analysis reveals 

alterations in histone acetyltransferase, histone methyltransferase, 

SET domain proteins and histone deacetylases in the 

alcoholic liver. Five genes upregulated include ATF2 (histone 

acetyltransferase), PRMT6 and SETD7 (histone methyltransferases), 

RPS6KA3 (kinase) and HDAC3 (histone deacetylase) 

whereas HDAC9 (histone deacetylase) was downregulated 

during ALD. Analysis of expression in isolated hepatocytes and 

Kupffer cells exhibits cell type specific regulation of expression. 

ATF2 was exclusively upregulated (2 fold) in alcohol exposed 

Kupffer cells while PRMT6 (3.2 fold) increased in hepatocytes. 

HDAC9, on the other hand, was down regulated (7 fold) in 

alcohol exposed hepatocytes. HDAC3, SETD7 and RPS6KA3 

were increased in KCs and hepatocytes. Inhibition of hsp90 

using 17-DMAG after chronic alcohol exposure significantly 

alleviated liver injury measured by reduced serum ALT and liver 

triglycerides. 17-DMAG altered expression of ATF2, PRMT6, 

HDAC3 and HDAC9 in the liver. Phosphorylation of ATF2 in 

LPS stimulated macrophages activates transcription of pro-inflammatory 

cytokines. 17-DMAG treatment after alcohol feeding 

prevented an increase in ATF2 expression (P


hsp90 impacts epigenetic chromatin modifying enzymes 

important in innate immune activation and lipid metabolism in 

a cell-specific manner in ALD. 

Disclosures: 

The following authors have nothing to disclose: Pranoti Mandrekar, Aditya 

Ambade, Arlene Lim 

1352 

Cyr61 protection of hepatocytes from alcohol-induced 

endoplasmic reticulum stress 

Sang Geon Kim, Tae Hyun Kim, Hyun Joo Kim; College of Pharmacy, 

Seoul National University, Seoul, Korea (the Republic of) 

The endoplasmic reticulum (ER), the major organelle responsible 

for the biosynthesis, folding, assembly and modification 

of soluble proteins and membrane proteins, senses cellular or 

extracellular stresses. Since the liver plays a key role in energy 

metabolism and toxicant detoxification as the portal organ, a 

variety of harmful stimuli, including imbalance of energy supply, 

alcohol intake, oxidative stress, and inflammatory mediators 

induces ER stress, which may facilitate hepatocyte dysfunction 

and injury. Cysteine-rich angiogenic protein 61 (Cyr61), a 

member of matricellular cysteine-rich protein (CCN) family, 

regulates cell survival, differentiation and inflammation. This 

study investigated whether Cyr61 inhibits alcoholic steatohepatitis 

(ASH), and if so, what the underlying mechanism is using 

the database of ASH patients, animal and hepatocyte models. 

Of the CCN family members, the transcript levels of CCN1 

encoding for Cyr61 were uniquely and markedly diminished 

in patients with alcoholic hepatitis. In the liver tissue or hepatocytes 

prepared from mice fed on Lieber-DeCarli alcohol diet for 

4 weeks or exposed to binge alcohol, Cyr61 expression levels 

were substantially decreased with reciprocal increases of fatty 

acid synthase and acetyl-CoA carboxylase. Cyr61 knockdown 

augmented ER stress in hepatocytes, as indicated by changes 

in ER stress markers, CHOP, Grp78, XBP-1 and IRE-1a. Conversely, 

overexpression of Cyr61 suppressed ER stress. Moreover, 

Cyr61 knockdown promoted PARP cleavage with the 

induction of CYP2E1. Cyr61 overexpression diminished ethanol-induced 

PARP-1 cleavage. Overall, our results demonstrate 

that Cyr61 repression by alcohol intake promotes hepatocyte 

injury through increase of ER stress, providing a novel link 

between Cyr61 and ER stress in the progression of ASH. 

Disclosures: 

Sang Geon Kim - Consulting: Nature’s Sunshine Products, Inc 

The following authors have nothing to disclose: Tae Hyun Kim, Hyun Joo Kim 

1353 

IL-13Rα1 signaling and M2 macrophages but not Th2 T 

cells drive progression of CCL 4 

-induce fibrosis 

Shih-Yen Weng 1 , Xiaoyu Wang 1 , Yong Ook Kim 1 , Leonard Kaps 1 , 

Yilang Tang 2 , Olena Molokanova 1 , Jeff R. Crosby 3 , Michael L. 

McCaleb 3 , Brombacher Frank 4 , Tobias Bopp 5 , Hans-Joerg Schild 5 , 

Ari Waisman 2 , Detlef Schuppan 1,6 ; 1 Institute of Translational 

Immunology, University Medicine, Johannes Gutenberg University, 

Mainz, Germany, Mainz, Germany; 2 Institute for Molecular 

Medicine, Mainz, Germany; 3 Isis Pharmaceuticals, Carlsbad, 

CA; 4 Institute of Infectious Disease and Molecular Medicine, Cape 

Town, South Africa; 5 Institute of Immunology and Research Center 

for Immunotherapy at UMC Mainz, Mainz, Germany; 6 Beth Israel 

Deaconess Medical Center, Boston, MA 

Background and aims: Liver fibrosis progression and regression 

are modulated by cells of the immune system. CD4 T cells 

regulate functional macrophages polarization by secreting 

cytokines such as IFNg, IL-12, IL-4 and IL-13. To investigate the 

roles of IL-4 and IL-13 in liver fibrosis development, systemic 

IL-4/IL-13 double KO mice (IL-4/IL13 -/- ) and T cell specific deletion 

of IL-4/IL-13 (IL-4/IL13 ∆CD4 ) and IL-4Rα (IL-4Rα ∆CD4 ) were 

generated and subjected to liver fibrosis analysis. Methods: 

Liver fibrosis progression was modeled by administration of 

oral CCL 4 

in increasing doses for 6 weeks. Antisense oligonucleotides 

(ASO) were injected intraperitoneally at 40mg/kg 3 

times per week during the 6 weeks of CCL 4 

treatment . Results: 

Histology analysis of IL-4/IL-13 -/- mice at 6 week of CCL 4 

treatment 

revealed significant reduction of collagen deposition indicated 

by less Sirius Red-stained area (50% decrease) in. This 

was accompanied by 6-fold reduction of procollagen α1(I) 

and α-SMA transcripts, and a decreased ALT indicating suppressed 

liver injury. IL-4/IL-13 -/- mice also showed an increase 

in CD4 and CD8 T cells, monocytes and macrophages/Kupffer 

cells as assessed by FACS. However, IL-4/IL-13 ΔCD4 mice and 

IL-4Rα ΔCD4 mice showed no significant difference in liver fibrosis 

compared to control mice. Therefore Th2 cells appear to 

play only a minor role in CCL 4 

-induced fibrosis, suggesting 

other fibrogenic cells responding to IL-4/13. IL-13Rα1 was 

previously characterized as an M2 macrophage marker. We 

therefore applied an ASO against IL-13Rα1 to CCL 4 

-treated 

mice. Compared to control ASO treated mice, liver fibrosis as 

assessed by collagen quantification and Sirius red morphometry, 

was significantlyy attenuated by this treatment. Conclusions: 

We demonstrate an important role of IL-4/IL-13 signaling 

in CCL 4 

-induced fibrogenesis. However, IL-4/13 producing 

Th2 T cells do not play a significant role in fibrosis progression, 

while M2 macrophages, responsive to IL-13, apparently produced 

by other sources, are central to fibrosis progression. This 

qualifies IL-13 and IL-13Rα1 as important targets for antifibrotic 

therapies. 

Disclosures: 

Jeff R. Crosby - Employment: ISIS Pharmaceuticals 

Michael L. McCaleb - Employment: Isis Pharmaceuticals; Stock Shareholder: Isis 


The following authors have nothing to disclose: Shih-Yen Weng, Xiaoyu Wang, 

Yong Ook Kim, Leonard Kaps, Yilang Tang, Olena Molokanova, Brombacher 

Frank, Tobias Bopp, Hans-Joerg Schild, Ari Waisman, Detlef Schuppan


1354 

Myofibroblastic conversion of portal fibroblasts and 

mesothelial cells isolated from adult mouse liver 

Ingrid A. Lua, Kinji Asahina; University of Southern California, 

Department of Pathology, Keck Scool of Medicine, Los Angeles, 

CA 

Purpose: Hepatic stellate cells (HSCs) are believed to be the 

major source of myofibroblasts in liver fibrosis; however, other 

cellular sources, such as portal fibroblasts (PFs) in the portal 

triad and mesothelial cells (MCs) on the liver surface, have also 

been suggested. It is unknown how these cells contribute to 

fibrogenesis due to insufficient availability of markers and isolation 

methods. Methods: We isolated HSCs, PFs and MCs from 

collagen1a1 (Col1a1)-GFP transgenic mouse livers by fluorescence-activated 

cell sorting (FACS) and identified their markers 

by microarray analysis. Primary HSCs, PFs and MCs were 

cultured in the presence of TGF-β1 or PDGF-BB to examine their 

myofibroblastic conversion and proliferation. Results: Immunohistochemistry 

showed that Col1a1-GFP mouse livers broadly 

express GFP in quiescent HSCs, PFs, and MCs. Combining 

vitamin A (VitA) lipid autofluorescence with GFP expression, 

we separated the VitA+ HSCs and VitA-GFP+ population from 

the Col1a1-GFP livers through FACS. We further subtracted 

glycoprotein M6A (GPM6A)+ MCs from the heterogeneous 

VitA-GFP+ population and obtained a PF-enriched fraction as 

VitA-GFP+GPM6A- cells. Microarray analysis identified selective 

expression of Reelin in HSCs. PFs exclusively expressed 

Ectonucleoside triphosphate diphosphohydrolase-2 (ENTPD2/ 

NTPDase2). Isolated PFs exhibited fibroblastic morphology 

without storing VitA lipids in culture. MCs showed a round 

morphology and expressed CD200, mesothelin, podoplanin, 

and uroplakin 1b. HSCs, PFs, and MCs that were isolated from 

normal Col1a1-GFP livers transformed into myofibroblasts that 

expressed α-smooth muscle actin induced by TGF-β1 in culture. 

TGF-β1 suppressed Cyclin D mRNA expression in PFs but not in 

HSCs and MCs. By contrast, PDGF-BB induced Cyclin D mRNA 

only in HSCs. Conclusion: HSCs, PFs, and MCs have the potential 

to differentiate into myofibroblasts, and their proliferation is 

regulated differently by TGF-β1 and PDGF-BB. 

Disclosures: 

The following authors have nothing to disclose: Ingrid A. Lua, Kinji Asahina 

1355 

Selective disruption of dynamin GTPase activity in HSC 

increases murine fibrogenesis in vivo and promotes HSC 

activation in vitro 

Qian Ding 1 , Ruisi Wang 1 , Thiago de Assuncao 1 , Meng Yin 2 , 

Sheng Cao 1 , Usman Yaqoob 1 , Richard Ehman 2 , Robert C. Huebert 

1 , Vijay Shah 1 ; 1 Gastroenterology Research Unit, Mayo Clinic, 

Rochester, MN; 2 Department of Radiology, Mayo Clinic, Rochester, 

MN 

Background: Dynamin-2 (Dyn2) is a large GTPase responsible 

for endocytosis. A point mutation of Dyn2 at lysine 44 to 

alanine disrupts dynamin GTPase activity (Dyn2K44A). Prior 

studies using a mouse we generated that conferred Dyn2K44A 

expression selectively in endothelial cells impaired angiogenesis 

in vivo by disrupting receptor tyrosine kinase endocytosis 

and signaling. We hypothesized that expression of Dyn2K44A 

in hepatic stellate cells (HSC) would similarly disrupt tyrosine 

kinase dependent HSC activation and fibrogenesis in vivo. 

Methods and Results: Dyn2K44A fl/fl mice were crossed with 

Collagen 1-Cre (Col1 Cre ) mice to generate offspring with HSC 

selective expression of Dyn2K44A (Col1 Cre /Dyn2K44A fl/fl ). 

Primary HSC isolates showed Dyn2K44A expression by realtime 

PCR. Col1 Cre /Dyn2K44A fl/fl mice and littermate controls 

were subjected to carbon tetrachloride (CCl 4 

) or olive oil vehicle 

for 6 weeks, or bile duct ligation (BDL) or sham operation 

for 3 weeks to induce fibrosis and then sacrificed. Contrary 

to our hypothesis, immunofluorescence for α-smooth muscle 

actin (α-SMA) and fibronectin (FN), and sirius red staining from 

fixed liver tissues was significantly greater in response to both 

BDL and CCl 4 

in Col1 Cre /Dyn2K44A fl/fl mice compared with 

littermate controls (~1.5-fold; p


exosome binding to HSC by RGD (p < 0.05) but not by RGE. 

Binding of PKH26-stained HSC-derived exosomes to PKH67- 

stained HSC, assessed over 12 hrs, was also dose-dependently 

inhibited by EDTA (0-500 μM; p < 0.05). Since these data suggested 

exosome-HSC interactions involve integrins and since 

all five αV integrins (β1, β3, β5, β6, β8) and two β1 integrins 

(α5, α8) are RGD-dependent, recipient HSC were transfected 

for 24 hrs with siRNA to αV or β1 and then incubated with 

PKH26-stained exosomes for 24 hrs. Each treatment alone was 

effective in blocking >95% exosome binding. Finally, binding 

of HSC-derived exosomes to recipient HSC was reduced by 

heparin but not chondroitin sulfate (100μg/ml) while exosomal 

miR-214-mediated suppression of CTGF 3’-UTR activity, accomplished 

by 24-hr co-culture of miR-214-transfected donor HSC 

with CTGF 3’-UTR luciferase reporter-transfected recipient HSC 

was reversed in the presence of heparin but not chondroitin 

sulfate (100μg/ml) (p < 0.05). Thus exosome-HSC interactions 

or downstream gene activity are heparin- or integrin-dependent, 

the latter of which requires expression of αV and/or β1 

subunits by recipient HSC. 

Disclosures: 


The following authors have nothing to disclose: Li Chen, Ruju Chen, Sherri Kemper 

1357 

Potential of connective tissue growth factor as a novel 

therapeutic target against hepatic fibrosis 

Yuki Makino, Hayato Hikita, Yugo Kai, Yasutoshi Nozaki, Tasuku 

Nakabori, Yoshinobu Saito, Satoshi Tanaka, Ryotaro Sakamori, 

Naoki Hiramatsu, Tomohide Tatsumi, Tetsuo Takehara; Department 

of Gastroenterology and Hepatology, Osaka University 

Graduate School of Medicine, Suita, Japan 

Background and aim: Connective tissue growth factor (CTGF) 

is a multifunctional matricellular protein reported to be up-regulated 

in the liver of patients with chronic liver diseases. However, 

the association of CTGF and liver fibrosis has not been 

fully studied. We analyzed the significance of CTGF and its 

potential as a therapeutic target in hepatic fibrosis. Methods/ 

Results: Using clinical resected livers of 93 patients under the 

approve of Research Ethics Committee, we revealed that CTGF 

expression levels in non-tumorous liver tissue increased along 

with the progression of fibrotic degrees of the liver and positively 

correlated with those of other fibrosis-related genes such 

as α-SMA, TGF-β1, and Col1a1. In a mouse experiment, CTGF 

was up-regulated in the liver 3 weeks after bile duct ligation 

(BDL). CTGF expression levels showed positive correlation with 

those of other fibrosis-related genes listed above. Immunochemistry 

of CTGF after BDL showed positive reaction both in parenchymal 

cells (PC) and non-parenchymal cells (NPC). Thus, we 

subsequently isolated PC and NPC from mice underwent BDL 

or sham, and compared respective CTGF expression levels 

between BDL and sham groups. CTGF was up-regulated both 

in PC and NPC in BDL groups. In vitro experiments revealed 

that treatment of TGF-β, an inducing factor of CTGF, increased 

CTGF expression and secretion not only in hepatocyte cell 

lines, Huh7 and PLC/PRF/5 cells but in hepatic stellate cell 

(HSC) cell lines, LX-2 cells. These findings indicated that both 

hepatocytes and HSC produce CTGF under the stimulation of 

BDL. We finally analyzed the potential of CTGF as a therapeutic 

target in liver fibrosis. In vitro CTGF treatment induced the 

activation and collagen production in LX-2 cells, suggesting 

CTGF has a promoting effect for liver fibrosis. We subsequently 

evaluated the degree of fibrosis after BDL in 3 strains of conditional 

CTGF knockout mice; HSC-specific CTGF deficient mice 

(GFAP-Cre+ CTGF flox/flox mice; CTGFΔHSC mice), hepatocyte-specific 

CTGF deficient mice (Alb-Cre+ CTGF flox/flox 

mice; CTGFΔHep mice), and poly IC-induced PC and NPC 

CTGF deficient mice (MX1-Cre+ CTGF flox/flox mice; CTG- 

FΔPC+NPC mice). As a result, compared with Cre- littermates, 

CTGF expression levels in the liver were significantly reduced 

to 55.8% and 27.7% in CTGFΔHep and ΔPC+NPC mice, 

respectively. No difference was observed in CTGFΔHSC mice. 

Liver fibrosis was attenuated only in CTGFΔPC+NPC mice, evidenced 

by reduced Collagen expression and Sirius Red-stained 

area. Conclusion: CTGF is produced from both PC and NPC 

and promotes hepatic fibrosis. CTGF is a promising therapeutic 

target in liver fibrosis. 

Disclosures: 






The following authors have nothing to disclose: Yuki Makino, Yugo Kai, Yasutoshi 

Nozaki, Tasuku Nakabori, Yoshinobu Saito, Satoshi Tanaka, Ryotaro Sakamori, 

Naoki Hiramatsu, Tomohide Tatsumi 

1358 

Exosome-mediated activation of TLR3 in stellate cells 

stimulates IL-17A production of γδ T cells in liver fibrosis 

of mice 

Wonhyo Seo, Hyuk-Soo Eun, So Yeon Kim, Seol-Hee Park, Jong- 

Min Jeong, Won-Mook Choi, Myung-Ho Kim, Won-IL Jeong; 

KAIST, Daejeon, Korea (the Republic of) 

Background: During liver injury, hepatocytes secrete numerous 

exosomes including diverse types of self-RNAs. Recently, 

self-noncoding RNA has been considered as an activator to tolllike 

receptor 3 (TLR3). However, the roles of hepatic exosome 

and its detection by TLR3 have not been fully understood in the 

pathogenesis of liver fibrosis. In the present study, we identified 

that exosome-induced activation of TLR3 in hepatic stellate cells 

(HSCs) stimulated production of interleukin-17A (IL-17A) in γδ 

T cells, subsequently leading to acceleration of liver fibrosis 

in mice. Methods: Liver injury and fibrosis was induced with 

single or 2-week injection of carbon tetrachloride (CCl 4 

) in wild 

type (WT) and TLR3-deficient (TLR3 -/- ) mice. Liver immune cells, 

HSCs and hepatocytes were freshly isolated for flow cytometry 

and in vitro experiments. Results: In acute and chronic liver 

injuries, increased IL-17A production was mainly detected in 

hepatic γδ T cells of WT mice, whereas those of TLR3 -/- mice did 

not show any increase of IL-17A production in both liver injuries. 

In addition, liver fibrosis of TLR3 -/- mice was significantly 

attenuated compared with WT mice. More interestingly, IL-17A 

producing γδ T cells were in close contact with activated HSCs, 

suggesting implication of HSCs and TLR3 in IL-17A production 

of γδ T cells. Furthermore, in vitro treatments of exosomes 

derived from CCl 4 

- or ethanol-treated hepatocytes significantly 

increased expression of IL-1β, IL-23 and IL-17A in HSCs, and 

remarkable increase of IL-17A production in γδ T cells was 

observed as they were co-cultured with exosome-treated WT 

HSCs or conditioned medium from TLR3-activated WT HSCs. 

However, all these finding were not detected as γδ T cells were 

co-cultured with exosome-treated HSCs of IL-17A -/- or TLR3 - 

/- 

mice. Using reciprocal bone marrow transplantation of WT 

and TLR3 -/- mice, we demonstrated TLR3 deficiency in HSCs 

contributed to decreased liver fibrosis and IL-17A production 

in γδ T cells. Conclusion: In liver injury, exosome-mediated activation 

of TLR3 in HSCs accelerates liver fibrosis by enhancing 

IL-17A production of γδ T cells, which might be associated 

with IL-17A production of HSCs by unknown self-TLR3 ligands


released from damaged hepatocytes. Therefore, TLR3 might be 

a novel therapeutic target for liver fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Wonhyo Seo, Hyuk-Soo Eun, So 

Yeon Kim, Seol-Hee Park, Jong-Min Jeong, Won-Mook Choi, Myung-Ho Kim, 

Won-IL Jeong 

Effects of ASK1 inhibitors on fatty acid metabolism 

1359 

Efficacy of an ASK1 Inhibitor to Reduce Fibrosis and 

Steatosis in a Murine Model of NASH is Associated with 

Normalization of Lipids and Hepatic Gene Expression 

and a Reduction in Serum Biomarkers of Inflammation 

and Fibrosis 

Satyajit Karnik 1 , Michael R. Charlton 2 , Li Li 1 , Michelle Nash 1 , 

Maisoun Sulfab 1 , David Newstrom 1 , Erik G. Huntzicker 1 , Dorothy 

French 1 , Zachary D. Goodman 3 , Tracy Shafizadeh 4 , Steve Watkins 

4 , David Breckenridge 1 , Daniel Tumas 1 ; 1 Biology Research, 

Gilead Sciences, Foster City, CA; 2 Gastroenterology and Hepatology, 

Mayo Clinic, Rochester, MN; 3 Center for Liver Diseases, 

Inova Fairfax Hospital, Falls Church, VA; 4 Metabolon Inc, West 

Sacramento, CA 

BACKGROUND & SIGNIFICANCE: ASK1 is a kinase that is activated 

by various stimuli including oxidative stress, TGF-β, and 

hyperglycemia. ASK1 induces apoptosis, fibrosis, and metabolic 

dysfunction by activating p38 and JNK1. We previously 

demonstrated that ASK1 inhibition is efficacious in preclinical 

models of NASH and liver fibrosis. GS-4997, a first-in-class, 

oral small molecule ASK1 inhibitor, is being investigated in a 

Phase 2 clinical trial in NASH. Here, we further characterize 

the efficacy of ASK1 inhibition in a murine model of NASH by 

evaluating hepatic gene expression, hepatic and serum lipids, 

and serum biomarkers and cytokines. METHODS: NASH was 

induced in male C57BL/6 mice by administration of a diet 

high in fat, cholesterol, and sugar for 240 days. Animals were 

subsequently treated with either placebo or a selective, small 

molecule inhibitor of ASK1 for 90 days (n=15/group). Endpoints 

included steatosis evaluated by morphometry, clinical 

pathology, liver hydroxyproline levels, liver and plasma lipids 

by chromatography, hepatic gene expression by microarray, 

serum levels cytokines and fibrosis biomarkers by Luminex and 

ELISA. RESULTS: ASK1 inhibition reduced hepatic fibrosis, steatosis, 

and insulin resistance. ASK1 inhibitor treated animals 

had increased lipid metabolism and decreased de novo lipogenesis 

vs. controls as evaluated by lipid profiling in liver. 

ASK1 inhibitor treated animals had a 26% decrease in lipogenesis, 

a 21% decrease in stearoyl-CoA desaturase, and a 

78% increase in delta-5 desaturase relative to placebo controls 

as reflected in the plasma lipidome (Table). Consistent with 

these effects, gene networks regulating fatty acid synthesis, 

lipid metabolism, and cholesterol biosynthesis were differentially 

expressed in ASK1 inhibitor treated animals vs. placebo 

controls. Serum levels of IL-6, and the fibrosis markers osteopontin, 

hyaluronic acid, and TIMP-1 were reduced by 50%, 

35%, 33% and 41%, respectively, in ASK1 inhibitor treated 

animals vs. controls. CONCLUSIONS: The therapeutic benefit 

of an ASK1 inhibitor in reducing hepatic steatosis and fibrosis 

was associated with normalization of fatty acid synthesis and 

lipid metabolism. These data enhance our understanding of the 

mechanisms of action of ASK1 inhibition and further support 

evaluation of the ASK1 inhibitor, GS-4997, in patients with 

NASH. 

Disclosures: 

Satyajit Karnik - Consulting: Monsoon Dx; Employment: Gilead Sciences 



Li Li - Employment: Gilead Sciences 

Michelle Nash - Employment: Gilead Sciences 

Erik G. Huntzicker - Employment: Gilead Sciences; Stock Shareholder: Gilead 

Sciences 

Dorothy French - Employment: Gilead Sciences; Stock Shareholder: Genentech 

- Roche 




David Breckenridge - Employment: Gilead Sciences 

Daniel Tumas - Employment: Gilead Sciences, Inc 

The following authors have nothing to disclose: Maisoun Sulfab, David Newstrom, 

Tracy Shafizadeh, Steve Watkins 

1360 

Endothelial Niche Derived Notch Signaling Pathway Balanced 

Liver Regeneration and Fibrosis in Distinct Ways 

Lin Wang; Hepatic Surgery, Fourth Military Medical University, 

Xi`an, China 

Abstract: Traumatic or toxic damage to the liver is often 

associated with liver regeneration and fibrosis. Liver sinusoidal 

endothelial niche has been addressed to modulate liver 

regeneration and fibrosis. Previously we found, Notch signaling 

was involved into the determination of liver sinusoidal 

endothelial cell (SEC) proliferation and differentiation by an 

angiocrine manner, and consequently influenced liver regeneration. 

However, the specific modulations and cell communications 

by endothelial niche-derived Notch signaling on 

liver reconstruction are still unknown. In this study, by using 

endothelial specific Notch Intracellular Domain (NICD) activated 

transgenic models (VeCad-CreERT-NICD) to persistently 

activate Notch signal, we firstly found SEC fenestration and 

sieve plates dramatically decreased both in vivo and in vitro. 

Meanwhile, sinusoidal perfusion and SEC endocytosis ability 

were significantly compromised compared to the control. 

Intriguingly, incapable hepatocyte proliferation, activated 

hepatic stellate cells (HSC) and liver malfunction were more 

remarkable in VeCad-CreERT-NICD mice. In 70% hepatectomy 

models, hepatocyte regeneration was largely blocked by NICD 

activation. By contrast, hepatic fibrosis induced by CCl 4 

injection 

was apparently aggravated in Notch activated models. 

Excitingly, in different co-culture systems, isolated primary SEC 

with loss of fenestrations by Notch activation disrupted wild 

type hepatocyte proliferation, but initiated HSC activation. To 

illustrate the underlying mechanisms, we detected Wnt2, which 

was considered as a mitogen of hepatocytes, was downregulated 

in VeCad-CreERT-NICD SEC. Overexpression of Wnt2 

successfully restored incapable hepatocyte proliferation in 

Notch activated mice. Besides, prominently increased ET1 was 

noticed in Notch activated SEC and potentially accelerated 

liver fibrosis in a Notch-ET1 dependent manner. Conclusion: 

We demonstrated Notch signal as a bi-directional switch in 

balancing hepatic regeneration and fibrosis through sinusoidal 

endothelial niche derived pro-regenerative Notch-Wnt2 versus 

pro-fibrotic Notch-ET1 pathways.


Disclosures: 

The following authors have nothing to disclose: Lin Wang 

1361 

Liver fatty acid-binding protein is required for Perilipin 

5 to restore lipid droplets in activated hepatic stellate 

cells 

Jianguo Lin 1 , Elizabeth P. Newberry 2 , Nicholas O. Davidson 2 , 

Anping Chen 1 ; 1 Pathology, Saint Louis University, St. Louis, MO; 

2 Medicine, Washington University in St. Louis, St. Louis, MO 

Hepatic stellate cell (HSC) activation is coupled to the loss of 

lipid droplets (LDs) which are specialized organelles composed 

of neutral lipids surrounded by perilipins (Plins), among which 

Plin5 plays a key role in regulating aspects of intracellular 

trafficking, signaling and cytoskeletal organization in hepatocytes. 

Recent work in Plin5 ko mice suggests a role in high fat 

diet induced hepatic lipotoxicity. We now show that high fat 

diet induced liver fibrosis is accompanied by >70% reduction 

in HSC Plin5 and that spontaneous activation of primary 

HSCs produces temporally coincident loss of Plin5 expression 

and LD depletion. Since modulating lipid content in HSCs is 

a suggested strategy for inhibiting HSC activation and treatment 

of hepatic fibrosis we asked whether exogenous Plin5 

expression in primary HSCs would reverse the activation phenotype 

and promote LD formation. Lentiviral Plin5 expression 

in primary wild type (WT) mouse HSCs restored LDs, increased 

lipid content and suppressed activation (~2-fold reduced procollagen 

expression), coincident with ~2 fold upregulation of 

both PPARgamma and L-Fabp expression. Since HSC specific 

expression of L-Fabp promotes FA accumulation and LDs and 

inhibits HSC activation in vitro, we next asked if L-Fabp expression 

is required for Plin5 rescue of the activated phenotype. 

Lentiviral Plin5 transduction of HSCs from L-Fabp –/– mice failed 

to restore lipogenic enzyme expression (FAS, SREBP1c) and 

showed significantly attenuated induction of PPARgamma (WT 

~5-fold induction vs KO ~2.5 fold). Lentiviral Plin5 transduction 

also failed to induce LD formation in HSCs from L-Fabp –/– mice 

and failed to reverse the activation phenotype (no change in 

aSMA or procollagen 1 mRNA, vs significant ~2-fold reduction 

in WT HSCs). To further verify the functional role of L-Fabp in 

Plin5 mediated pathways, we introduced adenoviral L-Fabp 

into HSCs from L-Fabp –/– mice, which completely rescued the 

ability of lentiviral Plin5 to restore LD formation and increase 

lipid content. Control, lacZ transduced HSCs from L-Fabp –/– 

mice failed to demonstrate increased lipid content or visible LDs 

after lentiviral Plin5 transduction. Taken together, our results 

indicate that Plin5 expression in HSCs plays a critical role in 

the activation phenotype both in vivo and in vitro. Furthermore, 

our findings demonstrate that HSC L-Fabp has a critical role in 

the pathways by which Plin5 modulates lipogenic gene expression, 

restoring LD formation and overall lipid content and in 

turn modulating HSC activation. 

Disclosures: 

The following authors have nothing to disclose: Jianguo Lin, Elizabeth P. Newberry, 

Nicholas O. Davidson, Anping Chen 

1362 

Deletion of Wntless in macrophages exacerbates liver 

fibrosis and the ductular reaction in chronic liver injury 

Katharine Irvine 1 , Andrew D. Clouston 1 , Antje Blumenthal 3 , Elizabeth 

E. Powell 1,2 ; 1 Centre for Liver Disease Research, The University 

of Queensland, Brisbane, QLD, Australia; 2 Department of 

Gastroenterology and Hepatology, Princess Alexandra Hospital, 

Brisbane, QLD, Australia; 3 UQ Diamantina Institute, The University 

of Queensland, Brisbane, QLD, Australia 

Introduction: Macrophages play critical roles in liver regeneration, 

fibrosis development and resolution. They are among the 

first responders to liver injury, and are implicated in orchestrating 

the fibrogenic response via multiple mechanisms. Macrophages 

are intimately associated with the activated hepatic 

progenitor cell (HPC) niche, or ductular reaction, that develops 

in parallel with fibrosis. Among the many macrophage-derived 

mediators implicated in liver disease progression, a key role 

for macrophage-derived Wnt proteins in driving pro-regenerative 

HPC activation towards a hepatocellular fate has been 

suggested. Wnt proteins, in general, however, have been 

associated with both pro- and anti-fibrogenic activities in the 

liver and other organs. We investigated the role of macrophage-derived 

Wnt proteins in fibrogenesis and HPC activation 

in murine models of chronic liver disease. Methods: We 

generated mice deficient in Wnt secretion from macrophages 

(MΦ) by deleting the Wls gene, which is essential for Wnt 

secretion, in LysM-Cre-expressing cells. Liver injury and fibrosis 

were induced by administration of thioacetamide (TAA, 12 

wks) in drinking water, or a choline-deficient ethionine-supplemented 

diet (CDE, 6 wks). Fibrosis and the ductular reaction 

were assessed histologically, and gene expression in whole 

liver and flow cytometry-sorted macrophages and HPC was 

assessed in MΦ-Wls-knockout mice and littermate controls. 

Results: Fibrosis and HPC activation were exacerbated in 

MΦ-Wls mice compared to littermate controls in TAA and CDE 

treated mice (TAA, p


1363 

Mast cells interact with proliferating cholangiocytes to 

activate hepatic stellate cells and promote fibrosis via 

TGF-β1 signaling during cholestatic injury 

Lindsey Kennedy 2 , Laura Hargrove 1 , Jennifer Owens 2 , Heather 

L. Francis 2,1 ; 1 Scott and White Memorial Hospital, Temple, TX; 

2 Research, Central Texas Veteran’s Health Care System, Temple, 

TX 

Background: Hepatic fibrosis is marked by activation of hepatic 

stellate cells (HSCs) and the release of TGF-β1. Cholestatic 

injury is a precursor to liver fibrosis and, following injury cholangiocytes 

acquire a neuroendocrine phenotype and interact 

with HSCs to promote fibrosis. Mast cells (MCs) infiltrate the 

liver following injury and release pro-fibrogenic factors including 

histamine that increases biliary proliferation and fibrosis. 

We tested the hypotheses that (i) MCs induce biliary proliferation, 

HSC activation and promote fibrosis and (ii) inhibition of 

MC-derived histamine decreases fibrosis via TGF-β1. Methods: 

To demonstrate that MCs promote fibrosis, wild-type (WT) and 

HDC -/- mice (characterized by lower MC count) were subjected 

to sham surgery or bile duct ligation (BDL) and injected with 

cultured MCs via tail vein. Further, WT and MDR2 -/- mice (a 

model of Primary Sclerosing Cholangitis characterized by 

increased MC count) were treated with cromolyn sodium to 

block MC-derived histamine. We collected liver blocks, serum, 

cholangiocytes and biliary supernatants. Biliary mass and proliferation 

were evaluated by immunohistochemistry for CK-19 

and PCNA, respectively. Fibrosis was evaluated in tissue sections 

by Sirius Red/Fast Green Collagen staining and by qPCR 

in total liver and cholangiocytes for α-SMA, fibronectin, collagen 

type 1a and TGF-β1. HSC activation was evaluated by 

qPCR in total liver and immunofluorescent staining in tissues 

for synaptophysin 9 (SYP9). TGF-β1 secretion was measured in 

serum from all groups. Downstream from TGFβ-1, we measured 

SMAD2 and SMAD3 by qPCR and western blotting in total 

liver and cholangiocytes. In vitro, cultured MCs were transfected 

with HDC shRNA to decrease histamine secretion and 

subsequently co-cultured with cholangiocytes or HSCs prior to 

measuring fibrosis markers, proliferation and TGF-β1 secretion. 

Results: Injected mast cells were found in close proximity to proliferating 

bile ducts. Biliary proliferation, fibrosis and HSC activation 

were increased in HDC -/- and BDL HDC -/- mice following 

MC injection. Inhibition of MC-derived histamine decreased 

biliary proliferation, fibrosis, TGF-β1 secretion and SMAD2/3 

expression in MDR2 -/- mice. In vitro, knockdown of MC HDC 

decreased (i) MC TGF-β1 secretion, (ii) biliary proliferation and 

fibrotic marker expression and (iii) HSC activation and TGF-β1 

secretion. Conclusion: MCs are recruited to proliferating cholangiocytes 

and promote HSC activation increasing fibrosis via 

TGF-β1/SMAD signaling. Inhibition of MC-derived histamine 

decreases fibrosis and regulation of MC mediators may be a 

therapeutic target for fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Lindsey Kennedy, Laura Hargrove, 

Jennifer Owens, Heather L. Francis 

1364 

Hepatocyte autophagy impairment promotes ductular 

reaction and fibrogenesis by hepatic stellate cells 

Youngmin A. Lee 1 , Luke A. Noon 1,2 , Tingfang Lee 1 , Marie-Luise 

Berres 3,6 , Fatemeh P. Parvin-Nejad 1 , Kemal M. Akat 4 , Hsin I 

Chou 1 , Varinder Athwal 1,7 , M. Isabel Fiel 5 , Ronald E. Gordon 5 , 

Scott L. Friedman 1 ; 1 Liver Diseases, Icahn School of Medicine at 

Mount Sinai, New York, NY; 2 Centro de Investigación Principe 

Felipe (CIPF), Valencia, Spain; 3 Department of Internal Medicine 

III, University Hospital, RWTH Aachen, Aachen, Germany; 4 Laboratory 

of RNA Molecular Biology, HHMI/Rockefeller University, 

New York, NY; 5 Department for Pathology, Icahn School of Medicine 

at Mount Sinai, New York, NY; 6 Department of Oncological 

Sciences, Icahn School of Medicine at Mount Sinai, New York, 

NY; 7 Institute of Human Development, Faculty of Medicine and 

Human Sciences, University of Manchester, Manchester, United 

Kingdom 

Background Ductular reaction (DR), an expansion of ductular 

progenitor-like cells, is often correlated with fibrosis in chronic 

liver disease and fulminant liver injury. Murine models in which 

autophagy, a homeostatic lysosomal pathway, is abrogated 

in hepatocytes by deletion of the autophagy related protein 7 

(Atg7) display a prominent DR with marked elevation of liver 

transaminases, steatohepatitis and fibrosis, with eventual tumorigenesis. 

Although hepatic stellate cells (HSCs) are associated 

with the DR, their interactions have not been well characterized 

and their relative contribution to fibrosis is uncertain. Our 

aim was to determine the contribution of ductular and HSCs 

to fibrosis when hepatocyte autophagy is impaired. Methods: 

Olig1-Cre:Atg7 F/F mice, which have selective hepatocyte 

depletion of the autophagy protein Atg7, were analyzed by 

immunohistochemistry (IHC), qRT-PCR and microarray for fibrogenic 

markers. Primary HSCs and ductular cells were isolated 

following collagenase perfusion and gradient purification. 

Enrichment of respective cell populations was confirmed by 

flow analysis and IHC. Gene expression was then analyzed by 

qRT-PCR. To identify fibrogenic cells in vivo, Olig1-Cre:Atg7 F/F 

mice were crossed to transgenic Col1a1-GFP mice, in which 

GFP expression is regulated by the Collagen1a1 promoter. 

Livers of Tg(Col1-GFP)cKO(Olig1-Cre:Atg7 F/F ) were analyzed 

by IHC for GFP as well as markers for ductular cells (EpCAM, 

CD133) and HSCs (desmin). Results: Olig1-Cre:Atg7 f/f mice 

develop extensive ductular reaction (Epcam + , CD133 + by IHC) 

and marked spontaneous fibrosis at 3 months, as assessed by 

sirius red staining (8.4 ± 2.4 vs 1.1±0.2 % fibrotic tissue area 

in controls), with significant fibrogenic gene induction in whole 

liver RNA based on qRT-PCR and array. Strikingly, collagen 

fibers were closely aligned with ductular cells (EpCAM + , Ck7 + ) 

that were also intimately associated with HSCs (desmin + ). 

HSCs and ductular cells isolated from Olig1-Cre:Atg7 F/F both 

expressed aSMA, a marker of activated HSCs, and Col1, 

whereas none was expressed in control livers. In transgenic 

Col1a1-GFP reporter mice with hepatocyte autophagy loss, 

GFP expression was greater in desmin positive cells than in 

cells expressing progenitor markers (EpCAM, CD133), establishing 

HSCs as the primary fibrogenic cells. Conclusion: 

Hepatic fibrosis is a prominent and novel feature of hepatocyte 

Atg7 KO mice. HSCs intimately associated with ductular cells 

are the main fibrogenic cell population. These findings help 

clarify the basis of fibrogenesis associated with the DR and 

establish an experimental model for elucidating cell-cell interactions 

in this response.


Disclosures: 











The following authors have nothing to disclose: Youngmin A. Lee, Luke A. Noon, 

Tingfang Lee, Marie-Luise Berres, Fatemeh P. Parvin-Nejad, Kemal M. Akat, Hsin 

I Chou, Varinder Athwal, M. Isabel Fiel, Ronald E. Gordon 

1365 

Hepatocyte- and hepatic stellate cell-specific deletion of 

liver fatty acid binding protein (L-Fabp) reveals divergent, 

cell-type specific roles in hepatic lipid trafficking 

and fibrogenesis 

Elizabeth P. Newberry 1 , Susan M. Kennedy 1 , Gianfranco Alpini 3 , 

Anping Chen 2 , Nicholas O. Davidson 1 ; 1 Washington University 

School of Medicine, St Louis, MO; 2 St. Louis University, St Louis, 

MO; 3 Central Texas Veterans Health Care System and Texas A&M 

HSC College of Medicine, Temple, TX 

Liver fatty acid binding protein (L-Fabp) modulates fatty acid 

(FA) trafficking, high fat diet-induced obesity, and steatosis. We 

previously showed that germline L-Fabp –/– mice have reduced 

FA uptake and incorporation into complex lipid in both hepatocytes 

and stellate cells (HSC), with attenuated diet-induced 

hepatic steatosis and fibrosis. Since L-Fabp is expressed in 

both hepatocytes and HSC, we examined its cell-specific role in 

lipid trafficking and fibrotic injury. Germline L-Fabp –/– mice fed 

a trans-fat, fructose (TFF) diet exhibit reduced steatosis and liver 

fibrosis, with a 70% decrease in Col1a1 and αSMA mRNAs. 

Hepatocyte-specific (Alb-Cre) L-Fabp deletors (Hep KO, >95% 

decreased L-Fabp mRNA) exhibit reduced hepatic TG (f/f, 560 

± 71mg/mg; Hep-KO, 284 ± 91; p=0.046), with decreased 

liver size and expression of Col1a1 and αSMA (~50%), suggesting 

that HSC activation in TFF-fed mice is linked to hepatocyte 

lipid accumulation. In contrast, HSC-specific (GFAP-Cre) 

L-Fabp deletion (HSC KO, ~75% reduction) did not affect liver 

size, hepatic steatosis or expression of Col1a1, but αSMA 

mRNA was reduced. Because GFAP Cre may target other cell 

types, we verified that WT cholangiocytes express undetectable 

L-Fabp. Freshly isolated HSCs from HSC-KO contain abundant 

lipid droplets, in contrast to germline L-Fabp –/– HSC which are 

lipid droplet depleted. Together these data indicate that TFF 

diet induced fibrotic injury is linked to hepatocyte TG content 

and suggest that lipid trafficking to HSCs may reflect FA compartmentalization 

in hepatocytes. In a distinct model of liver 

injury, carbon tetrachloride (CCl 4 

) administration produced 

worse injury in L-Fabp –/– mice vs C57BL/6J controls (ALT WT, 

666 ± 194 IU/L; KO, 1247 ± 179), with increased hepatocyte 

proliferation and enhanced expression of fibrogenic genes, 

yet no difference in fibrosis (WT, 2.6 ± 0.5%; KO, 2.4 ± 0.3, 

n=6). We then used HSC- and Hep- KOs to understand how 

L-Fabp deletion exacerbates CCl 4 

-induced liver injury. CCL 4 

- 

treated HSC KO mice exhibited higher serum ALT (f/f, 733 

± 306 IU/L; HSC-KO, 1450 ± 270, p=0.09) and hepatocyte 

proliferation (f/f, 7.2 ± 3.1 % BrdU+; HSC-KO 16.6 ± 1.6, 

p=0.02) with ~3-fold increased Col1a1 and αSMA expression. 

In contrast, CCl 4 

-treated Hep KO mice exhibited no difference 

in ALT (548 ± 186 IU/L) or hepatocyte proliferation 

versus controls. Together these data indicate that HSC L-Fabp 

expression plays a protective role against CCl 4 

injury, whereas 

hepatocyte L-Fabp promotes TFF diet-induced steatotic injury. 

Furthermore, the findings point to an unanticipated, cell specific 

role for L-Fabp in high fat diet induced versus toxin-mediated 

liver injury. 

Disclosures: 

The following authors have nothing to disclose: Elizabeth P. Newberry, Susan M. 

Kennedy, Gianfranco Alpini, Anping Chen, Nicholas O. Davidson 

1366 

Fibrous network composition and cell heterogeneity 

modulate long-range force transmission by cells in 

matrices 

Robert N. Kent, Jude D. Han, Rebecca G. Wells; Gastroenterology, 


Long-range force transmission is implicated in the large-scale 

architectural rearrangements occurring in fibrotic diseases like 

pulmonary fibrosis and liver cirrhosis. It was previously shown 

that matrix organized in a fibrous network is required and 

that collagen becomes aligned and compacted as part of this 

process. We hypothesized that the characteristics of the fibrous 

network as well as the heterogeneity of the cell population 

modulate long-range force transmission between cells. Fibroblasts 

(murine 3T3) were cultured as spheroids by suspension 

in a hanging droplet over 5 days. These were seeded in pairs 

500-1000 μm apart on thick gels of either denatured collagen 

(gelatin) or type I collagen (Col1) containing varying concentrations 

of type III collagen (Col3) or plasma fibronectin (pFN). 

Heterotypic spheroids containing varying ratios of primary rat 

hepatic stellate cells (HSCs) and hepatocytes were cultured in 

hanging droplets for 3 days and seeded on Col1 gels. Once 

seeded, spheroids were incubated or imaged via time-lapse 

microscopy for 24 hours and then fixed. Collagen alignment 

and compaction were visualized using second harmonic generation 

imaging, and the extent of alignment was quantified by 

calculating the anisotropy index in ImageJ (NIH). Gels without 

spheroids showed minimal collagen organization or alignment 

by second harmonic generation imaging. Col1 gels with spheroids 

showed marked alignment, contraction, and compaction 

of collagen along the axis between spheroids, with migration of 

cells along the compacted fibrils. Spheroids of 3T3 fibroblasts 

seeded on gelatin substrates showed minimal organization, 

suggesting that collagen cross-linking is required for cell-mediated 

network reorganization. On substrates consisting of 60% 

Col1 and 40% Col3, 3T3 spheroids generated significantly 

less fiber alignment when compared to spheroids on pure Col1 

substrates (p < 0.01). Col1 gels containing pFN (10% v/v), 

however, yielded a significant increase in alignment relative to 

pure Col1 (p < 0.05). Spheroids of primary HSCs reorganized 

the fibrous network such that the anisotropy index was significantly 

larger than the no-spheroid control (p < 0.05). The inclusion 

of hepatocytes in ratios of 1:20 and 1:1 hepatocyte:HSC 

yielded even greater network anisotropies than spheroids of 

HSCs alone (p < 0.01, 0.05). Collagen cross-linking, Col3 and 

pFN content, and heterogeneity of cell populations modulate 

long-range force transmission in fibrous networks. Determining 

the role of matrix cross-linking and cell heterogeneity in matrix 

reorganization may contribute to understanding the underlying 

mechanisms driving bridging in liver cirrhosis. 

Disclosures: 

The following authors have nothing to disclose: Robert N. Kent, Jude D. Han, 

Rebecca G. Wells


1367 

Dual combination therapy directed against lysyl oxidase-like 

2 (LOXL2) and apoptosis signal-regulating 

kinase 1 (ASK1) potently inhibits fibrosis and portal 

hypertension in a new mouse model of PSC-like liver 

disease 

Naoki Ikenaga 1 , Susan B. Liu 1 , Zhen-Wei Peng 1 , Andrew E. 

Greenstein 2 , Dorothy French 2 , Victoria Smith 2 , Yury Popov 1 ; 

1 Gatroenterology, Beth Israel Deaconnes, Boston, MA; 2 Gilead 

Sciences, Inc, Foster City, CA 

BACKGROUND/AIMS: We have previously characterized 

novel LOXL2 and ASK1 inhibitors as effective monotherapies 

for liver fibrosis in vitro and in vivo. They impact distinct, 

non-overlapping pro-fibrogenic signal transduction pathways, 

with LOXL2 mediating collagen cross-linking and ASK1 promoting 

hepatocyte injury and fibrogenic stellate cell activation 

via the p38 and JNK pathways. Here, the therapeutic efficacy 

of combined anti-LOXL2 and anti-ASK1 agents was evaluated 

in a mouse model of biliary fibrosis and portal hypertension. 

METHODS: We developed an improved BALBc.Mdr2-/- mouse 

model resembling human primary sclerosing cholangitis (PSC) 

with rapidly progressive fibrosis and early-onset portal hypertension 

(Ikenaga et al, 2015). 6 weeks of ASK1 inhibitor (GS- 

444217, 0.15% in diet), anti-LOXL2 monoclonal antibody 

(AB0023 mAb/GS-607601, 30mg/kg/week i/p), or their 

combination were administered to 6 weeks old BALB/c.Mdr2- 

/- mice with pre-established fibrosis (n=9-11/group). Portal 

venous pressure (PVP) was measured via direct cannulation of 

the portal vein with a micro-tip pressure monitor at the end of 

the study. Liver fibrosis was evaluated by histology and biochemical 

determination of collagen. RESULTS Both anti-LOXL2 

and anti-ASK1 mono-therapies significantly improved histological 

signs of fibrosis, with reduced hepatic collagen deposition 

(by 37 and 38% of hydroxyproline, respectively) in BALB/c. 

Mdr2-/- mice compared to placebo control group (p


sues and accelerates regression of CCl 4 

-induced fibrosis by 

stimulating expression of matrix metalloproteinase (MMP)-13 

and MMP-9. A subsequent study has identified opioid growth 

factor receptor-like 1 (OGFRL1) as a G-CSF-induced novel 

bone marrow cells-derived factor. To establish the molecular 

bases for future therapeutic application, we examined, in the 

present study, tissue distribution of endogenous OGFRL1 and 

the therapeutic effects of exogenous OGFRL1 administration. 

We also characterized the functions of OGFRL1 by generating 

its null mice. Methods: Total RNA was extracted from various 

organs/tissues of mice, and expression levels of Ogfrl1 gene 

were quantified using real time RT-PCR. Human bone marrow 

mesenchymal stem cells were infected with recombinant 

OGFRL1-expressing lentiviruses and injected into the spleen of 

CCl 4 

-treated nude mice. Hepatic expression of Mmp-13 and 

Mmp-9 mRNAs was examined by real time RT-PCR, and liver 

regeneration was estimated by BrdU uptake into hepatocytes. 

Ogfrl1 gene targeting was conducted using the CRISPR/Cas9 

system with a guide RNA set in the first exon of Ogfrl1 gene. 

Results: Endogenous OGFRL1 was expressed predominantly in 

neural tissues and, to the lesser extent, in hematopoietic organs 

such as bone marrow and spleen. Among the peripheral blood 

cells, monocytes and granulocytes were the major sources of 

OGFRL1. Liver tissues contained a small but significant amount 

of Ogfrl1 mRNA, which was decreased following repeated 

CCl 4 

injections. When fibrotic mice underwent partial hepatectomy, 

expression levels of endogenous Ogfrl1 gene were 

further declined. An intrasplenic injection of OGFRL1-expressing 

cells stimulated hepatic expression of MMP-13 and MMP-9, 

and increased the BrdU uptake into hepatocytes in fibrotic liver 

after partial hepatectomy. It also caused a remarkable increase 

in gene expression of cyclins and other cell cycle-related factors 

as well as that of AFP. OGFRL1 null mice exhibited no abnormalities 

in their developmental process or liver tissue architecture. 

However, mitosis of hepatocytes was delayed following 

partial hepatectomy in OGFRL1-null mice compared with wild 

type littermates. Conclusions: OGFRL1 plays a critical role in 

hepatocyte proliferation, and administration of OGFRL1-expressing 

cells could be a potential regeneration therapy for 

advanced liver fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Takayo Yanagawa, Hideaki Sumiyoshi, 

Sachie Nakao, Masato Ohtsuka, Minoru Kimura, Yousuke Chiba, Shintaro 

Satake, Tadashi Moro, Hiromi Chikada, Akihide Kamiya, Yutaka Inagaki 

1370 

Interleukin 15 regulates transforming growth factor 

β1-induced collagen production in hepatic stellate cells 

by controlling a network of suppressors of collagen 

transcription 

Kohtaro Ooka 3 , Huanzi Han 1 , Holger Fey 1 , Costica Aloman 1 , Jingjing 

Jiao 2 ; 1 Division of Gastroenterology/Liver Diseases (MC716), 

University of Illinois at Chicago, Chicago, IL; 2 University of Texas 

MD Anderson Cancer Center, Houston, TX; 3 Medicine, University 

of Illinois at Chicago, Chicago, IL 

Background & Aim: Interleukin 15 (IL15) signaling is unique 

in that IL15 binds to the high affinity IL15 receptor a subunit 

(IL15Rα) which it shuttles to the cell surface and stimulates 

opposing cells through the β/γ receptor complex. Our group 

has identified a protective role for IL15 signaling in murine models 

of liver fibrosis using IL15Rα knockout mice (IL15RαKO). 

In addition to the established effects of IL15 on maintaining 

natural killer (NK) cell homeostasis, a cell type well known to 

limit fibrogenesis, a direct anti-fibrotic effect of IL15 signaling 

on radio-resistant liver cells has also been observed. We aim 

to understand in detail the mechanism of the anti-fibrotic effect 

of IL15 signaling on hepatic stellate cells (HSC), the major type 

of fibrogenic cells in the liver,. Methods: HSCs were isolated 

from wild type (WT) and IL15RαKO mice by gradient centrifugation 

followed by depletion of contaminating leukocytes and 

endothelial cells. Purity of HSC was validated by the absence 

of mRNA for CD31 and CD45. HSC from WT and IL15RαKO 

were assessed by quantitative real time PCR (qPCR) and western 

blot to measure fibrogenic markers and well-known negative 

regulators of collagen transcription. Results: IL15RαKO 

HSCs display enhanced fibrogenesis both in the steady-state 

and under transforming growth factor β1 (TGF-β1) stimulation 

as shown by qPCR of collagen 1A1 (COL1A1), collagen 

3A1 (COL3A1) and platelet-derived growth factor receptor 

(PDGFR). Treatment with the IL15/IL15Rα complex reduced 

expression of COL1A1 and alpha smooth muscle actin (αSMA) 

in IL15RαKO HSCs and reduced the magnitude of αSMA 

expression induced by TGF-β1. Expression of several negative 

regulators involved in the collagen expression pathway including 

TP53, serine-threonine kinase receptor-associated protein 

(STRAP) and ski-like (SKIL) were lower in IL15RαKO HSCs and 

were further down-regulated by TGF-β1 treatment. Conclusion: 

IL15 signaling regulates suppressors of collagen transcription 

to have a direct anti-fibrotic effect on HSCs. Our study highlights 

a possible therapeutic effect of IL15 and IL15/IL15Rα in 

liver fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Kohtaro Ooka, Huanzi Han, 

Holger Fey, Costica Aloman, Jingjing Jiao 

1371 

NLRP3 inflammasome modulation of hepatic stellate cell 

activation: role in liver fibrosis 

Alexander Wree 1,2 , Matthew D. McGeough 1 , Davide Povero 1 , 

Akiko Eguchi 1 , Hal M. Hoffman 1 , Ariel E. Feldstein 1 ; 1 Pediatrics, 

UCSD, La Jolla, CA; 2 Internal Medicine III, University Hospital, 

RWTH-Aachen, Aachen, Germany 

Background: NLRP3 inflammasome activation has been 

increasingly recognized as an important mechanism of liver 

damage. Our group has recently demonstrated that isolated 

NLRP3 inflammasome activation is sufficient to induce liver 

inflammation and fibrosis (Hepatology 2014 Mar). Here we 

tested the hypothesis that the NLRP3 inflammasome in hepatic 

stellate cells (HSCs) can directly modulate their activation and 

contribution to liver fibrosis. Methods: An immortalized human 

HSC line (LX-2) was incubated with the NLRP3 inflammasome 

inducers, LPS (as initial priming signal) and ATP (as activating 

signal). Additionally, primary HSCs were isolated from 

Nlrp3 D301NneoR/+ knock-in mice on a C57BL/6 background 

which were crossed to C57BL/6.Cg-Tg(Cre/Esr1)–Estrogen 

Receptor Cre (CreT) mice allowing for temporal control of 

NLRP3 activation. In a third set of experiments, mutant Nlrp3 

was activated in the aforementioned mice in vivo for over 16 

weeks. Activation of the NLRP3 inflammasome and HSCs was 

assessed via fluorescent imaging techniques and mRNA analysis. 

Results: LX-2 cells incubated with LPS (1ug/ml, over 3h) 

showed increased mRNA levels of Nlrp3 (1.5 fold compared 

to control), Incubation with ATP (5mM) resulted in an additional 

increase (2.0 fold). Collagen-1 mRNA levels were only 

raised in LX-2 cells when incubated with LPS and ATP (1.5 

fold compared to control or LPS only). Fluorescently labeled 

active Caspase 1 was present in LX-2 cells when incubated 

with LPS + ATP. The NLRP3 inflammasome was induced in 

vitro in cultured primary HSCs via 4OH-tamoxifen treatment. 

Immunofluorescence for F-actin showed cytoskeleton reorga-


nization as early as 6h after 4OH-tamoxifen incubation with 

mRNA levels of Nlrp3, alpha smooth muscle actin and tissue 

inhibitor of metalloproteinase 1 being increased. Activation of 

the NLRP3 inflammasome in Nlrp3 D301N/+ /CreT mice over 16 

weeks resulted in hepatomegaly while tamoxifen-injected control 

mice showed regular liver weights (liver to body weight % 

- 6.1% ± 0.32% vs. 4.6% ± 0.03%, p < 0.05). Analysis of liver 

histology revealed increased inflammation with neutrophilic 

infiltration and increased mRNA levels of myeloperoxidase 

(fold-change to WT 52.8 ± 13.75, p


by the same parameters as above. We conclude that simultaneous 

targeting of peripheral CB 1 

R and iNOS by novel hybrid 

inhibitors offers improved efficacy and safety in the pharmacotherapy 

of liver fibrosis. 

Disclosures: 

Resat Cinar - Employment: National Institutes of Health; Patent Held/Filed: 

National Institutes of Health 

George Kunos - Patent Held/Filed: US Patent 

The following authors have nothing to disclose: Malliga Iyer, Ziyi Liu, Tony Jourdan, 

Zongxian Cao, Katalin Erdelyi, Grzegorz Godlewski, Gergo Szanda, Jie 

Liu, Pal Pacher, Kenner Rice 

1375 

Proof of liver regeneration after resolution of fibrosis by 

siRNA against HSP47 encapsulated in vitamin A-coupled 

liposome employing CCl 4 

mice engineered with 

Sox9-Cre ERT2 /ROSA-YFP genes 

Naoki Fujitani 1 , Akihiro Yoneda 2 , Yuya Murakami 1 , Miyuki 

Nishimura 1 , Miyono Miyazaki 3 , Keiko Kajiwara 3 , Kenjiro 

Minomi 3 , Yasuaki Tamura 2 , Yoshiro Niitsu 1 ; 1 School of Medicine, 

Department of Molecular Exploration, Sapporo Medical University, 

Sapporo, Japan; 2 Department of Molecular Therapeutics, Center 

for Food and Medical Innovation, Institute for the Promotion of 

Business-Regional Collaboration, Sapporo, Japan; 3 Hokkaido Laboratory, 

Nitto Denko Corporation, Sapporo, Japan 

Background and Aim We have previously shown that hepatic 

fibrosis could be resolved by the administration of siRNA 

against HSP47 encapsulated in vitamin A-coupled liposome 

(VA-lip-siRNA HSP47) in various liver cirrhosis rodent models 

(Sato et al. Nat. Biotechnol. 2008). However, a real clinical 

benefit of such anti-fibrosis modality would be a recovery of 

impaired liver function with tissue regeneration. We therefore, 

in the present study intended to demonstrate that fibrotic liver 

is capable to regenerate after resolution of fibrosis, employing 

bile duct ligation (BDL) model rats, dimthylnitrosoamine (DMN) 

model rats and carbon tetrachloride (CCl 4 

) model of transgenic 

mice carrying Sox9-Cre ERT2 /ROSA-YFP genes. Methods BDL 

and DMN models of rats were established as reported earlier 

(Sato et al. Nat. Biotechnol. 2008). Transgenic mice with 

Sox9-Cre ERT2 /ROSA-YFP genes were produced by repeated 

crossing of Sox9-Cre ERT2 mice onto ROSA-YFP mice obtained 

from The Jackson Laboratory. Fibrosis was induced with the 

intraperitoneal administration of CCl 4 

for 10 weeks. VA-lipsiRNA 

HSP47 was formulated as described earlier (Sato et al. 

Nat. Biotechnol. 2008). Immuno-stainings for Sox9, αSMA, 

EpCAM, YFP, HNF4α, BrdU and PCNA were carried out using 

each specific antibodies. Results and discussion In the liver of 

BDL rats, EpCAM positive; progenitors were localized at portal 

area and were surrounded by deposited collagen and αSMA 

positive cells, activated stellate cells (aHSC) as revealed by 

immune-staining. In the liver of DMN rats which were administrated 

with VA-lip-siRNA HSP47 for 4 times, both aHSC and 

progenitors markedly diminished, and PCNA staining onto 

HNF4α positive hepatocytes was significantly increased as 

compared with that of control rats, suggesting a concept that 

in the cirrhotic liver progenitors surrounded by aHSCs forming 

niche like structure underwent differentiation into hepatocytes 

upon destruction of niche structure with administration 

of VA-lip-siRNA HSP47. To verify this concept, fate-tracing of 

progenitor after resolution of fibrosis was conducted with Sox9- 

Cre ERT2 /ROSA-YFP mice. When the CCl 4 

mice were injected 

with tamoxifen, followed by the treatment of VA-lip-siRNA 

HSP47 for 10 times, initiating at 10 th week of CCl 4 

administration, 

and YFP expression was examined at 13 th week of 

CCl 4 

administration, HNF4α positive hepatocytes were clearly 

immune-stained for YFP. Thus, present study demonstrates that 

cirrhotic liver is capable of undergoing regeneration with our 

anti-fibrosis treatment even under the condition of continuous 

exposure to hepatotoxic agent. 

Disclosures: 

Miyuki Nishimura - Grant/Research Support: Nitto Denko 

The following authors have nothing to disclose: Naoki Fujitani, Akihiro Yoneda, 

Yuya Murakami, Miyono Miyazaki, Keiko Kajiwara, Kenjiro Minomi, Yasuaki 

Tamura, Yoshiro Niitsu 

1376 

Oral hepatotropic DPP4 inhibitors attenuate NASH and 

bilary fibrosis in part through macrophage modulation 

Xiaoyu Wang 1 , Shih-Yen Weng 1 , Yong Ook Kim 1 , Thomas Klein 3 , 

Detlef Schuppan 1,2 ; 1 Institue of Translational Immunology, Mainz, 

Germany; 2 Division of Gastroenterology, Boston, MA; 3 Boehringer 

Ingelheim Pharma, Biberach an der Riss, Germany 

Background and aims: Non-alcoholic steatohepatitis (NASH) is 

characterized by steatosis, lobular inflammation and progressive 

parenchymal fibrosis. Glucagon like peptide 1 (GLP-1) 

analogues are proven drugs to treat type 2 diabetes and insulin 

resistance. GLP-1 levels are increased by agents that inhibit cell 

surface dipeptidyl peptidase-4 (DPP-4) which is ubiquitously 

expressed and rapidly inactivates secreted GLP-1. Here, we 

studied the effect of two hepatotropic DPP4-inhibitors on liver 

inflammation and fibrosis in models of NASH and biliary fibrosis. 

Methods: Linagliptin and Sitagliptin were administered 

daily by oral gavage to Mdr2KO mice and to C57BL/6mice 

fed a methionine and choline deficient (MCD) diet for 6 weeks. 

Hepatic fibrosis was assessed by morphometric analysis of 

Sirius red stained collagen and measurement of hydroxyproline 

content. Hepatic inflammation and fibrosis were assessed by 

semiquantitative immunohistochemistry. Fibrosis and inflammation 

related transcript levels were measured by quantitative 

real-time polymerase chain reaction (qPCR). Ex vivo analysis 

of hepatic inflammatory cells was done by FACS. Results: In 

the MCD model, both Linagliptin and Sitagliptin lead to a significantly 

attenuated hepatic fat accumulation a reduction of 

transcript levels of SREBP-1c, FAS and LPL, lowered serum ALT, 

AST, ALP and LDH and significantly suppressed fibrosis and 

inflammation related transcripts (aSMA, procollagen α1(I), 

TIMP-1, TGFβ1, MMP-13, CD68, CCL3 and TNFα) compared 

to vehicle-treated controls. This was accompanied by a significant 

decrease of collagen area (Sirius red), αSMA, procollagen 

type III, CD68, F4/80, Caspase 3, as determined by semiquantitive 

immunohistochemistry. Boih DPPIV-inhibitors significantly 

decreased hepatic CD11b + Ly6C + high (proinflammatory monocytes/macrophages) 

and total F4/80 + cells (macrophages, 

Kupffer cells) compared to mice on the MCD diet without treatment. 

In nonsteatotic, spontaneously fibrotic Mdr2KO mice 10 

mg/kg/day of Linagliptin significantly decreased procollagen 

α1(I), TGFβ1, TIMP-1, MMP-8 transcript levels, and increased 

putatively anti-fibrotic MMP-9 and -13. Conclusion: In mice fed 

the MCD diet for 6 weeks, the DPPIV-inhibitors significantly 

decreased inflammation and apoptosis. In both the MCD and 

Mdr2KO model Linagliptin and Sitagliptin mildly decreased 

liver injury and fibrosis through inhibition of inflammatory and 

apoptosis pathways and via a decrease of proinflammatory 

and profibrotic monocytes-macrophages. 

Disclosures: 

Thomas Klein - Employment: Boehringer Ingelheim 




The following authors have nothing to disclose: Xiaoyu Wang, Shih-Yen Weng, 

Yong Ook Kim


1377 

Branched-chain amino acids prevent hepatic fibrosis 

and the development of liver tumors by suppressing 

TGF-β signaling 

Kai Takegoshi 1 , Masao Honda 1 , Hikari Okada 1 , Riuta Takabatake 

1 , Takayoshi Shirasaki 1 , Naoto Matsuzawa 1 , Toshinari 

Takamura 1 , Takuji Tanaka 2 , Shuichi Kaneko 1 ; 1 Kanazawa University, 

Kanazawashi, Japan; 2 Tohkai Cytopathology Institute, Gifu, 

Japan 

BACKGROUND & AIMS Oral supplementation with branchedchain 

amino acids (BCAAs) in patients with liver cirrhosis 

potentially suppresses the incidence of hepatocellular carcinoma 

and improves event-free survival. However, the detailed 

mechanisms by which BCAAs act on hepatic fibrosis have not 

been elucidated fully. METHODS BCAAs were administered to 

atherogenic and high-fat (Ath&HF) diet-induced nonalcoholic 

steatohepatitis (NASH) model mice and platelet-derived growth 

factor C transgenic (Pdgf-c Tg) mice. Liver histology, tumor 

incidence, and GeneChip expression profiles were evaluated. 

To investigate the mechanisms of the effect of BCAAs, in vitro 

studies were performed using a human stellate cell line (Lx-2), 

rat liver progenitor cell-like cells (WB-F344), mouse primary 

hepatic stellate cells (MHSC), and mouse primary hepatocytes 

(MPH). RESULTS Mice fed the Ath&HF diet for 30 weeks developed 

liver fibrosis, whereas mice fed the Ath&HF diet containing 

BCAAs showed markedly reduced fibrosis. After 60 weeks, 

73.5% of mice fed the Ath&HF diet developed liver tumors, 

while BCAAs reduced tumor incidence to 30.8%. BCAAs also 

prevented liver fibrosis of Pdgf-c Tg mice and reduced tumor 

incidence. Gene expression analysis demonstrated the significant 

resolution of pro-fibrotic genes, pro-oncogenic genes, cancer 

stem cell markers (EpCAM and CD90), and impaired lipid 

metabolism gene expression by BCAA supplementation. In 

vitro, in stellate cells, BCAAs restored the transforming growth 

factor (TGF)-β1-stimulated expression of pro-fibrotic genes and 

inhibited the trans-differentiation of MHSC to myofibroblast-like 

cells. In hepatocytes, BCAAs prevented TGF-β1 and palmitate 

co-stimulated apoptosis signaling and restored impaired fatty 

acid β oxidation gene expression in MPH. Moreover, in progenitor 

cells, BCAAs reduced the TGF-β1-stimulated spheroid 

formation of WB-F344 cells. We found BCAAs inhibited TGF-β 

signaling by preventing the nuclear localization of the histone 

acetylating transcription factor NF-Y. Knockdown of NF-YA 

by small interfering RNA attenuated the TGF-β1-stimulated 

expression of p-AKT and p-Smad2/3. CONCLUSIONS BCAA 

supplementation prevents hepatic fibrosis and the development 

of liver tumors in the NASH mouse model and Pdgf-c Tg mice, 

possibly by suppressing TGF-β signaling through an interaction 

with NF-Y. These results highlight a new mechanism of the 

anti-fibrotic and anti-tumor effect of BCAAs in the liver and 

could be utilized for the treatment of chronic liver disease. 

Disclosures: 







The following authors have nothing to disclose: Kai Takegoshi, Masao Honda, 

Riuta Takabatake, Takayoshi Shirasaki, Naoto Matsuzawa, Toshinari Takamura, 

Takuji Tanaka 

1378 

Membrane type I-matrix metalloprotease-cleaved collagens 

derived from hepatic stellate cells play critical role 

in liver regeneration after partial hepatectomy in rats 

Akihiro Yoneda 1,2 , Miyuki Nishimura 2 , Naoko Urushihara 2,3 , 

Tomoko M. Sudo 2,3 , Kenjiro Minomi 2,3 , Yasuaki Tamura 1 , Yoshiro 

Niitsu 2 ; 1 Department of Molecular Therapeutics, Center for Food & 

Medical Innovation, Institute for the Promotion of Business-Regional 

Collaboration, Hokkaido University, Sapporo, Japan; 2 Department 

of Molecular Target Exploration, School of Medicine, Sapporo 

Medical University, Sapporo, Japan; 3 Hokkaido Laboratory, Nitto 

Denko Corporation, Sapporo, Japan 

Background and Aims: The liver is well known to exhibit a 

remarkable ability to regenerate after partial heptatectomy 

(PHx). Liver regeneration is regulated by various events such as 

the remodeling of extracellular matrices (ECMs) and proliferation 

of hepatic cells stimulated by growth factors and cytokines. 

However, the precise molecular mechanism remains controversial. 

Since hepatic stellate cells (HSCs) are main producer of 

ECMs and also hepatic growth factor (HGF) in the damaged 

liver, HSCs has been postulated to play an important role in 

liver regeneration. We have recently demonstrated that membrane 

type I-matrix metalloprotease (MT1-MMP) of activated 

HSCs cleaved collagens secreted from themselves to expose 

RGD motif which interacted with integrins, thereby transduced 

survival signal and sustained their proliferation in an autocrine 

manner (Birukawa et al. J. Biol. Chem. 2014). The purpose of 

this study was to explore the possibility that MT1-MMP-cleaved 

collagens stimulate liver regeneration after PHx in a paracrine 

manner. Methods: Proliferations of hepatic cells after 70% PHx 

in rats were examined by immunohistochemistry and flow cytometric 

analysis. Hepatocytes isolated from EGFP-transgenic 

rats were co-cultured with activated HSCs. In vivo gene silencing 

system was performed using vitamin A-coupled liposome 

reported previously (Sato et al. Nature Biotechnol. 2008). 

Results: Proliferation of hepatocytes was coincide with activation 

of HSCs during the early phase of liver regeneration 

after 70% PHx. The proliferative hepatocytes localized in the 

vicinity of activated HSCs. Co-culture system demonstrated 

that activated HSCs stimulated the proliferation of hepatocytes 

while gene silencing of MT1-MMP or Hsp47, collagen-specific 

chaperone, in activated HSCs completely inhibited their 

proliferation. In vivo silencing of MT1-MMP or Hsp47 genes 

also resulted in remarkable suppression of liver regeneration 

after 70% PHx in rats. MT1-MMP-cleaved collagen I and HGF 

induced hepatocyte proliferation via the engagement of integrin 

b1 with c-Met tyrosine kinase receptor followed by the phosphorylation 

of c-Met and activation of focal adhesion kinase. 

We further found that proliferation of Sox9+ hepatic progenitor 

cells (HPCs) in close proximity to activated HSCs was transiently 

detected during the late phase of liver regeneration after 

70% PHx. Co-culture system demonstrated that activated HSCs 

induced the proliferation of HPCs while gene silencing of MT1- 

MMP or Hsp47 in activated HSCs completely inhibited their 

proliferation. Conclusion: MT1-MMP-cleaved collagens from 

HSCs play a pivotal role in liver regeneration after PHx. 

Disclosures: 

Miyuki Nishimura - Grant/Research Support: Nitto Denko 

The following authors have nothing to disclose: Akihiro Yoneda, Naoko Urushihara, 

Tomoko M. Sudo, Kenjiro Minomi, Yasuaki Tamura, Yoshiro Niitsu


1379 

Selective antibody targeting of lysyl oxidase-like 2 

(LOXL2) suppresses hepatic fibrosis progression and 

accelerates its reversal via crosslinking-dependent and 

independent mechanisms 

Naoki Ikenaga 1 , Susan B. Liu 1 , Zhen-Wei Peng 1 , Shuhei Yoshida 1 , 

Deanna Sverdlov 1 , Satyajit Karnik 2 , Amanda Mikels-Vigdal 2 , Victoria 

Smith 2 , Detlef Schuppan 1 , Yury Popov 1 ; 1 Gatroenterology, 

Beth Israel Deaconess Medical Center, Boston, MA; 2 Gilead Sciences, 

Inc, Foster City, CA 

BACKGROUND/AIMS: LOXL2 plays a role in collagen cross-linking 

and epithelial cell differentiation. We studied the role of 

LOXL2 in collagen cross-linking and hepatic progenitor cell 

(HPC) differentiation, and the therapeutic efficacy of anti-LOXL2 

mAb on liver fibrosis progression/reversal in mice. METHODS: 

Anti-LOXL2 antibody (AB0023 mAb, 30mg/kg), control antibody 

(M64, 30mg/kg) or placebo was administered i.p. 

twice a week during thioacetamide (TAA)-induced fibrosis progression 

(delayed treatment, week 6 to 12 of TAA) or during 

recovery (1-12 weeks off TAA). Therapeutic efficacy in biliary 

fibrosis was tested in BALB/c.Mdr2-/- and 3,5-diethoxycarbonyl-1,4-dihydrocollidine(DDC)-fed 

mice. Collagen cross-linking 

and fibrosis were assessed by histological and biochemical 

methods. HPC differentiation was studied in primary EpCAM(+) 

cells in vitro. RESULTS: LOXL2 was virtually absent from healthy 

but strongly induced in fibrotic livers, with predominant localization 

within fibrotic septa. Delayed anti-LOXL2 treatment 

of pre-established TAA-induced fibrosis suppressed collagen 

crosslinking, and histological signs of bridging fibrosis in the 

anti-LOXL2-treated group improved, with a 53% reduction in 

connective tissue deposition by morphometry. During recovery, 

anti-LOXL2 mAb promoted fibrosis regression, histological 

signs of fibrotic septa remodeling (splitting and thinning), 

and a 45% decrease in connective tissue area (p=0.021) at 

early recovery stages (4 weeks). In Mdr2-/- and DDC-induced 

models of biliary fibrosis, anti-LOXL2 mAb achieved significant 

antifibrotic efficacy but did not affect collagen cross-linking. 

Instead, ductular reaction and proliferation was suppressed 

in mice with LOXL2 inhibition, whereas hepatocyte replication 

increased. The LOXL2-blocking antibody had a profound effect 

on primary EpCAM(+) HPC in vitro, promoting their differentiation 

towards a hepatocyte lineage while inhibiting ductal cell 

lineage commitment. CONCLUSIONS: LOXL2 mediates collagen 

cross-linking and fibrotic matrix stabilization during liver 

fibrosis, and regulates hepatic progenitor cell differentiation. A 

therapeutic anti-LOXL2 antibody inhibits the progression of liver 

fibrosis and promotes fibrosis reversal. 

Disclosures: 

Satyajit Karnik - Employment: Gilead Sciences 

Victoria Smith - Employment: Gilead Sciences Inc 




Yury Popov - Grant/Research Support: Gilead Sciences, Inc, Takeda 

The following authors have nothing to disclose: Naoki Ikenaga, Susan B. Liu, 

Zhen-Wei Peng, Shuhei Yoshida, Deanna Sverdlov, Amanda Mikels-Vigdal 

1380 

IL-22 enhances TGF-beta pro-fibrotic function in hepatic 

stellate cells in a p38/MAPK dependent manner. 

Thomas Fabre 1,2 , Manuel Flores Molina 1,2 , Naglaa H. Shoukry 1,3 ; 

1 Centre de recherche du CHUM, Montrèéal, QC, Canada; 2 Département 

de microbiologie, immunologie et infectiologie, Université 

de Montréal, Montréal, QC, Canada; 3 Département de Médecine, 

Université de Montréal, Montréal, QC, Canada 

Background & Hypothesis: Activation of hepatic stellate 

cells (HSCs) is a key event in liver fibrosis, characterized by 

enhanced extracellular matrix (ECM) production and altered 

degradation. The immune system may modulate activation of 

HSCs through different cytokines. We have previously demonstrated 

that IL-17A enhances the expression of profibrotic genes 

through upregulation of the TGF-β receptor on hepatic stellate 

cells in a JNK-dependent manner (Fabre, Journal of Immunology 

2014). IL-22 is another Th17 enigmatic cytokine, from the 

IL-10 family, with both pro- and anti-inflammatory properties. 

IL-22 deficient mice develop high levels of hepatic inflammation 

during acute injury compared to their wild type littermates. 

In contrast, IL-22 is also elevated in the sera of patients with 

liver cirrhosis and carcinoma. However, in a mouse model 

of hepatitis B, IL-22 indirectly induces fibrosis by recruiting 

the pro-fibrotic inflammatory Th17 cells. We hypothesized that 

IL-22 may modulate activation and induction of the fibrogenic 

process in HSCs. Methods & Results: The human HSC line 

LX2 and primary human HSCs were stimulated with increasing 

doses of IL-22 and compared to TGF-β- and PBS- treated 

cells as positive and negative controls, respectively. IL-22 did 

not induce activation of HSCs. However, IL-22 enhanced the 

response of HSCs to suboptimal doses of TGF-β as observed by 

strong induction of alpha-smooth muscle actin (-SMA), collagen 

type I (COL1A1) and tissue inhibitor of matrix metalloproteinase 

(TIMP-I). IL-22 stimulation, in contrast to IL-17A, did not 

enhance cell surface expression of TGF-β-RII. To characterize 

the cellular mechanisms by which IL-22 enhances the TGF-β 

response we performed RNA-seq on primary human HSCs. 

However, pre-treatment of HSCs with IL-22 led to increase 

phosphorylation of SMAD2/3 in response to suboptimal doses 

TGF-β. Gene expression analysis revealed an increased in activation 

of p38/MAPK activity related pathways in IL-22 with 

TGF-β lo - as compared to PBS- or TGF-β lo treated cells. Activity 

was confirmed at the protein level and was associated with 

increased phosphorylation of SMAD2/3. Chemical inhibition 

of p38 significantly reduces HSCs activation in response to 

IL-22 with TGF-β lo . Conclusion: Our results suggest that IL-22 

enhances TGF-β signalling in a p38/MAPK dependent manner 

leading to increased activation of HSCs. 

Disclosures: 

The following authors have nothing to disclose: Thomas Fabre, Manuel Flores 

Molina, Naglaa H. Shoukry 

1381 

MeCP2 exerts global control over the myofibroblast 

transcriptome and reveals new regulators of fibrosis. 

EVA MORAN-SALVADOR, Pier P. Paoli, Graham R. Smith, Agata 

Page, Rachel M. Howarth, Fiona Oakley, Jelena Mann, Derek 

Mann; Institute of Cellular Medicine, Newcastle University, Newcastle 

upon Tyne, United Kingdom 

Background: Hepatic stellate cells (HSCs) are considered 

the central extracellular matrix (ECM)-producing cells in the 

wound-healing response process driven by a persistent liver 

injury. Previous studies in our laboratory identified MeCP2 

(methyl-CpG binding protein 2) as the epigenetics master reg-


ulator of HSCs activation in liver fibrogenesis. Nowadays, 

the epigenetic machinery embraces an extensive network of 

non-coding RNAs (ncRNAs), such as long non coding RNAs 

(lncRNAs) and microRNAs (miRNAs). The transcription of 

ncRNAs establishes new regulatory systems affecting chromatin 

structure and gene expression. The aim of this study was to 

identify potential targets and a novel ncRNA regulatory network 

controlled by MeCP2 in HSCs. Materials and methods: 

RNA from Wt (n=3) and MeCP2-null (n=3) mice HSCs was 

subjected to a microarray based profiling analyses (Arraystar 

Mouse LncRNA Microarray V2.0 platform) and to a small RNA 

next-generation sequencing (NGS) (Illumina platform). Results: 

Using the microarray platform we found 161 upregulated and 

83 downregulated lncRNAs in MeCP2-null HSCs. We focused 

our attention to Gm3893 and AK080187; their deregulated 

expression was confirmed by qPCR and correlated with the 

expression of their associated gene transcripts (cytokines Ccl- 

21b/c and IL11ra2 receptor and the profibrogenic marker 

Acta2, respectively). Pathway analysis based on KEGGs 

database denoted that most of 284 downregulated mRNAs 

in MeCP2-null HSCs were significantly enriched at DNA replication 

and cell cycle pathways, while most of 124 upregulated 

mRNAs were associated with pathways involved in the 

immune system and metabolism. Most importantly, MeCP2- 

null HSCs presented a robust downregulation in five members 

of the MCM protein complex and a remarkable decrease in 

other components of the replisome multisubunit complex. The 

resulting altered pattern of mRNAs also emphasized the downregulation 

of the hyaluronan synthase 2 (Has2) upon loss of 

MeCP2 in HSCs. In liver, Has2 was predominantly expressed 

in RNA of HSCs and its protein expression increased over 

the HSCs transdifferentiation process. In primary HSCs, small 

interfering RNA for Has2 decreased the gene expression of 

ECM proteins. Of note, miRNA-328-3p pinpointed from the 

14 significantly altered miRNA identified by NGS analyses 

as has been proved to target and reduce the hyaluronic acid 

receptor CD44 expression. Conclusions: Our findings successfully 

evidence a vast ncRNA network controlled by MeCP2 

and highlight the complex role that MeCP2 elicits in the HSC 

transdifferentiating process. 

Disclosures: 

The following authors have nothing to disclose: EVA MORAN-SALVADOR, Pier P. 

Paoli, Graham R. Smith, Agata Page, Rachel M. Howarth, Fiona Oakley, Jelena 

Mann, Derek Mann 

1382 

Cartilage oligomeric matrix protein participates in the 

pathogenesis of liver fibrosis 

Fernando Magdaleno 1,2 , Elena Arriazu 2 , Marina Ruiz de Galarreta 

2 , Yu Chen 1 , Xiadong Ge 1,2 , Laura Conde de la Rosa 2 , Natalia 

Nieto 1,2 ; 1 Pathology, University of Illinois at Chicago, Chicago, 

IL; 2 Medicine, Icahn School of Medicine at Mount Sinai, New 

York, NY 

BACKGROUND & HYPOTHESIS: Liver fibrosis is characterized 

by excessive accumulation of extracellular matrix (ECM) proteins, 

mainly fibrillar collagen-I, as a result of persistent liver 

injury. Cartilage oligomeric matrix protein (COMP) is a glycoprotein 

typically found in the ECM of skeletal tissue. Increased 

COMP expression has been associated with fibrogenesis in 

systemic sclerosis, lung fibrosis and chronic pancreatitis. Since 

COMP is expressed in cirrhotic livers and it is significantly 

induced in hepatocellular carcinoma we hypothesized that 

COMP could induce fibrillar collagen-I protein deposition and 

participate in matrix remodeling contributing to the pathophysiology 

of liver fibrosis. METHODS: Chronic thioacetamide (TAA) 

administration (4 mos) and carbon tetrachloride (CCl 4 

) injection 

(1 mo) were used as models to induce liver fibrosis in 

wild-type (WT) littermates and in global Comp -/- mice. Controls 

received water or mineral oil, respectively. To dissect the mechanism 

for COMP signaling, in vitro experiments were carried 

out with primary rat hepatic stellate cells (HSCs) and qPCR, 

gelatin zymography or western blot analysis were performed. 

To determine if physical interaction occurred between COMP 

and collagen-I and how this conditioned collagen-I cleavage 

by matrix metalloproteinases (MMPs), in vitro reconstituted systems 

were used followed by immunoprecipitation and immunoblotting. 

RESULTS: COMP expression was detected in livers 

from control WT littermates and was up-regulated in response 

to chronic TAA- or CCl 4 

-induced liver injury. TAA-treated or 

CCl 4 

-injected Comp -/- mice showed less liver damage (ALT and 

AST activities, necrosis, inflammation and hepatocyte ballooning 

degeneration) and fibrosis compared to their matching 

control WT littermates. Challenge of HSCs with recombinant 

COMP (rCOMP) up-regulated intra- and extracellular collagen-I 

protein expression and increased MMP2, 9 and 13. However, 

rCOMP neither affected TGFβ protein, a well-known pro-fibrogenic 

factor, nor altered Tnfα nor tissue inhibitor of metalloproteinase-1 

(Timp1) mRNAs. Moreover, COMP showed major 

ability to bind collagen-I; yet, the binding did not preclude from 

cleavage by MMP1, the MMP with the highest affinity for collagen-I 

cleavage. Last, we identified that rCOMP significantly 

up-regulated collagen-I protein expression in HSCs via CD36 

receptor binding and activation of the MEK1/2-pERK1/2 

signaling pathway. CONCLUSION: These results suggest that 

COMP contributes to liver fibrosis by up-regulating collagen-I 

protein synthesis but not its cleavage by MMP1; thus, resulting 

in significant collagen-I deposition. 

Disclosures: 

The following authors have nothing to disclose: Fernando Magdaleno, Elena 

Arriazu, Marina Ruiz de Galarreta, Yu Chen, Xiadong Ge, Laura Conde de la 

Rosa, Natalia Nieto 

1383 

IL-10-producing regulatory B cells from chronic hepatitis 

B patients suppressed CD4+T cells proliferation and 

enhanced regulatory T cells function 

Li-sha Cheng, Yun Liu, Wei Jiang; Gastroenterology, Zhongshan 

hospital, Shanghai, China 

Background & aims: Chronic hepatitis B (CHB) infection is a 

leading cause of liver fibrosis, cirrhosis and even hepatocellular 

carcinoma. Although hepatitis B virus (HBV) itself is non-cytopathic, 

both HBV-related liver damage and viral control are 

immune-mediated. There were abundant evidences to believe 

that regulatory T (Treg) cells play pivotal roles in the immunopathogenesis 

of HBV-related liver diseases. Nowadays, a 

new subset of IL-10-producing B cells has been shown to play 

vital roles in modulating autoimmune diseases. Here, we evaluated 

the role of regulatory B (Breg) cells in the pathogenesis 

of HBV-related liver fibrosis (HBV-LF) and assess their impact 

on the proliferation of CD4+T cells the immunoregulatory 

effects on Treg cells. Methods: A total of 67 treatment-naïve 

patients who were diagnosed as chronic HBV (CHB) and 

25 healthy donors took part in the study. Firstly, we determined 

the changes in number and function of Breg cells in 

CHB patients compared with that of healthy controls. For the 

in-vitro experiments, purified CD4+T cells were cultured alone 

or with autologous Breg cells and their proliferation response 

was determined by the thymidine method. Simultaneously, to 

elucidate the exact effects of Bregs on Tregs and the potential 

methanism, human Breg and Treg cells were co-cultured in the


stimulation with HBcAg, anti-IL-10 and anti-TGF-β antibody. 

Results: B cells and Breg cells were both enriched in patients, 

Breg cells frequencies had a close relationship with viral load, 

liver inflammation and degree of fibrosis. Stimultaneously, we 

demonstrated the phenotype of these Breg cells: memory B cells 

(CD24 hi CD27 + B cell)mature B cells(CD24 int CD38 int B cell) and 

transitional B cells (CD24 hi CD38 hi B cell), and the phenotype 

was pridominantly characterized as CD19 + CD24 hi CD38 hi B 

cell. In the co-culture of Breg cells from CHB patients with autologous 

CD4 + T cells, the proliferation capacity of CD4 + T cells 

was significantly reduced in a dose-dependent manner compared 

with controls. In addition, Breg cells from patients were 

found to have further enhanced effect on the proliferation of 

Treg cells as well as the release of IL-10 and TGF-β cytokines 

than such cells from controls. Furthermore, blockage of IL-10 

and CTLA-4 but not TGF-β decreased the inhibitory effect of 

Breg cells on Treg cells. The regulatory function of Breg cells on 

Treg cells was dependent on direct cell-cell contact as well as 

IL-10 but not TGF-β-dependent manner. Conclusions: Our data 

revealed that elevated Breg cells, which can inhibit the proliferation 

of CD4+T cells and regulate Treg cells immunitymaybe 

an indicator for the development of HBV-related liver fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Li-sha Cheng, Yun Liu, Wei Jiang 

1384 

SNAP-23 Heterozygous Mice Are Resistant To Experimental 

Fibrosis 

Haleigh B. Eubanks, Jessica R. Goree, Michel Fausther, Jonathan 

A. Dranoff; Gastroenterology and Hepatology, University of 

Arkansas for Medical Sciences, Little Rock, AR 

Background. Upon liver injury, quiescent hepatic stellate cells 

(HSC) become “activated” and converted to myofibroblasts. 

Hepatic stellate cell derived myofibroblasts possess characteristics 

such as increased proliferation capability and collagen 

synthesis. Several molecular targets have been identified 

in liver fibrosis. Most current therapies are aimed towards 

inhibiting or impeding accumulation of fibrogenic cells or 

deposition of extracellular matrix (ECM) proteins. However, 

the general molecular mechanism and proteins that function 

in aiding excessive ECM secretion remain to be elucidated. 

SNARE (soluble N-ethlmaleimide sensitive fusion attachment 

protein receptor) proteins have been proposed to be a conserved 

group of machinery that mediate intracellular trafficking 

and exocytosis. We found that HSC lack the t-SNARE protein, 

synaptosomal associated protein-25 (SNAP-25), but express a 

related protein, SNAP-23. Specific Aims. We are testing the 

hypothesis that SNAP-23 is essential for matrix release by HSC 

myofibroblasts by testing whether mice lacking one allele of the 

SNAP-23 gene are resistant to experimental fibrosis. (Of note, 

the SNAP-23 knockout mouse is embryonically lethal). Methods. 

Wild type and SNAP-23 heterozygous mice were treated 

with CCl4 or olive oil (control) for two (early fibrosis) and six 

weeks (late fibrosis/early cirrhosis). Fibrosis in liver tissue was 

assessed by Trichrome and Sirius red staining. Assessment of 

the expression of collagen, tissue inhibitor of metalloproteinase-1 

(TIMP-1), and alpha-smooth muscle actin (α-SMA) was 

determined by qRT-PCR. At the protein level, immunofluorescence 

and western blot were used to assess α-SMA secretion. 

Results. SNAP-23 +/- mice receiving CCl4 were significantly 

less fibrotic than WT mice; moreover, no SNAP-23 +/- mice 

were cirrhotic at the six week time point. Collagen quantification 

showed similar amounts of collagen content in WT and 

SNAP-23 +/- mice. While there was no difference in collagen 

or alpha-SMA mRNA expression between WT and SNAP-23 

+/- mice, SNAP-23 +/- mice evidenced decreased collagen 

and alpha-SMA protein. Conclusion. Single allelic deletion of 

SNAP-23 protected mice from experimental fibrosis/cirrhosis, 

likely via inhibition of exocytosis of matrix proteins. The reduction 

in alpha-SMA protein in heterozygote mice may reflect 

impaired HSC survival when the SNARE complex is disrupted. 

Disclosures: 

The following authors have nothing to disclose: Haleigh B. Eubanks, Jessica R. 

Goree, Michel Fausther, Jonathan A. Dranoff 

1385 

GP38/PODOPLANIN marks a novel subset of progenitor 

cells in chronic liver injury 

Henrike Julich 1 , Christoph Eckert 1 , Yong Ook Kim 2 , Hannes 

Borchardt 1 , Niklas Klein 1 , Eva-Carina Heier 1 , Thomas Tschernig 3 , 

Detlef Schuppan 2 , Veronika Lukacs-Kornek 1 ; 1 Department of 

Medicine II, Saarland University, Homburg, Germany; 2 Institute 

of Translational Immunology and Research Center for Immunotherapy, 

Institute of Translational Immunology, University Medical 

Center, Johannes Gutenberg University, Mainz, Germany; 3 Insitute 

of Anatomy and Cell Biology, Saarland University, Homburg, 

Germany 

BACKGROUND/AIMS: Gp38/podoplanin + stromal cells present 

in lymphoid organs play central role in the formation and 

reorganization of the extracellular matrix providing the three 

dimensional scaffold for immune responses. Additionally, 

gp38 + stromal cells are crucial in the functional regulation of T 

cells during immune responses (V. Lukacs-Kornek Nat Immunol 

2011 12(11):1096-104). Gp38 + cells have been identified 

during embryogenesis and more recently in human livers of primary 

biliary cirrhosis. Since, little is known about the function 

of these cells, we aimed to study the phenotypic and functional 

characteristics of gp38 + cells during chronic liver inflammation. 

METHODS: Gp38 + stromal cells have been analyzed in models 

of biliary fibrosis (MDR2 KO), CCL 4 

induced liver fibrosis 

(both during progression and regression) and in a model of 

steatohepatitis (methionine choline deficient (MCD) diet). Stromal 

cells were analyzed via flow cytometry and via confocal 

microscopy. Moreover, various subsets of stromal cells were 

sorted with high purity (>95%) for gene expression analyses 

and gp38 + stromal cells were tested functionally in in vitro 

assays. RESULTS: Gp38 + stromal cells are present in the healthy 

liver and significantly expand in all three models of liver injury. 

During regression of inflammation and fibrosis, the presence 

of these cells returns to the levels of healthy animals. Gp38 + 

cells localized primarily in the portal area and occasionally 

imposed as round ductular structures. Using multicolor FACS 

analysis, two subsets of gp38 + stromal cells could be identified: 

gp38 hi and gp38 low . The gp38 hi cells expressed multiple 

mesenchymal markers while the gp38 low cells could be further 

divided using CD133 and EPCAM labeling. Importantly, the 

gp38 low subset showed surface markers similar to liver progenitor 

(oval) cells, which was confirmed by qPCR array analyses. 

Importantly, gp38 low cells displayed inflammatory genes 

during liver injury different from classical oval cells identified as 

CD45 - gp38 - CD133 + cells. CONCLUSIONS: 1.Gp38 + cells are 

present in healthy liver. 2. Gp38 + stromal cells expand during 

liver injury. 3. Multiple subsets of gp38 + stromal cells could 

be distinguished: the gp38 low cells represent a novel oval cell 

subset with a distinct inflammatory gene expression compared 

to classical oval cells which after further characterization may 

represent a novel therapeutic target for liver inflammation and 

fibrosis. Studies were supported by the Alexander von Humboldt 

Foundation – Sofja Kovalevskaja Award 2012 to VLK, 

and an ERC Advanced Grant to DS.


Disclosures: 




The following authors have nothing to disclose: Henrike Julich, Christoph Eckert, 

Yong Ook Kim, Hannes Borchardt, Niklas Klein, Eva-Carina Heier, Thomas 

Tschernig, Veronika Lukacs-Kornek 

1386 

Characterization of obesity-related modifications in 

hepatic stellate cells 

Guanhua Xie 1 , Marzena Swiderska-Syn 1 , Mariana V. Machado 1 , 

Mark Jewell 1 , Richard T. Premont 1 , Gregory A. Michelotti 1 , Raffaele 

Teperino 2 , Anna Mae Diehl 1 ; 1 Division of Gastroenterology, 

Duke University Medical Center, Durham, NC; 2 Institute of Experimental 

Genetics (IEG) - HDC, Neuherberg, Germany 

Background: Obesity is an independent risk factor for 

advanced fibrosis in older NAFLD patients. Paradoxically, ob/ 

ob mice with genetic leptin deficiency are obese but protected 

from liver fibrosis. Hedgehog-responsive myofibroblasts (MF) 

derived from hepatic stellate cells (HSC) are the major fibrogenic 

cells in obesity-related cirrhosis. The anti-obesogenic hormone, 

leptin, promotes transdifferentiation of HSC into MF. 

Aim: To determine if genetic leptin deficiency induces inherent 

differences in HSC that account for the “fibrosis-protected” 

liver phenotype in ob/ob mice. Methods: Primary HSCs were 

isolated from wild type (WT) and ob/ob mice by in situ pronase/collagenase 

perfusion followed by histodenz density gradient 

centrifugation. To exclude the possibility of hepatocyte 

contamination, some experiments were repeated in ultrapure 

HSCs obtained by additional retinoid-based FACS sorting. 

mRNA and/or protein expression of myofibroblastic markers 

and Hedgehog (Hh) signaling components/targets were compared 

by qRT PCR and FACS analysis when HSCs were either 

freshly isolated or cultured for 7 days to induce myofibroblastic 

transition. Scratch and Brdu assays were used to examine the 

migration and proliferation of HSCs. Studies were repeated 

with HSC harvested from leptin-treated ob/ob mice, and with 

cultured ob/ob HSC that were treated with leptin or Sonic 

Hedgehog (SHh) ligand. Results: Ob/ob HSCs were less myofibroblastic 

(e.g., ≥ 50% reduced mRNA and protein expression 

of α smooth muscle actin, collagen 1α1, and platelet-derived 

growth factor) and had inhibited Hh pathway activity (e.g., 

less patched 1, Gli1, Gli2, and SHh at mRNA and/or protein 

level) compared to WT HSCs, both when quiescent and culture-activated. 

In contrast, a 2-fold greater elevated expression 

of quiescent HSCs markers (peroxisome proliferator-activated 

receptor γ and glial fibrillary acidic protein) were found in 

ob/ob HSCs. Functional analysis by Brdu assay and migration 

assay also showed that ob/ob HSCs were significantly 

less proliferative and migratory (p


would use human liver, or a biologically relevant 3D in vitro 

model of human liver cells grown on an extracellular matrix 

(ECM) derived from healthy or cirrhotic liver tissue. Aim: The 

aim of this study was to develop a rapid protocol for the decellularisation 

of small samples of healthy and cirrhotic human 

liver and demonstrate repopulation with cultured human liver 

cell lines, namely hepatocarcinoma (HepG2) and hepatic stellate 

(LX2) cells. Methods: Liver tissue cubes (125mm 3 ) were 

dissected from human livers unsuitable for transplantation or 

explanted liver. The decellularisation of the liver scaffolds was 

completed within 3hrs (healthy) and 5hrs (cirrhotic) of agitation 

in decellularization solutions i.e. detergents/enzymes. The 

decellularization efficiency was determined by immunohistochemistry 

for ECM components and residual DNA, scanning 

electron microscopy, biomechanics, as well as DNA and ECM 

protein quantification. The liver scaffolds were repopulated by 

HepG2 and LX2 for up to 14 days. RESULTS: This innovative 

protocol resulted in liver scaffolds with a preserved 3D structure 

and ECM composition, while DNA and cellular residues 

were successfully removed. The cirrhotic liver structure was 

maintained after decellularization and the biomechanical properties 

were remarkably different when comparing healthy liver 

scaffolds with cirrhotic scaffolds. Liver scaffolds were progressively 

repopulated for up to 14 days with LX2 and HepG2 cells 

which showed remarkable viability, motility and proliferation 

associated with remodelling effects on the surrounding ECM. 

Notably, the expression of several genes and proteins involved 

in liver fibrosis and cancer was different between the healthy 

and cirrhotic 3D-system. CONCLUSION: This is the first report 

describing an efficient protocol to completely decellularize 

human liver scaffolds obtained from healthy and cirrhotic liver. 

This decellularization protocol maintained the natural 3D-structure 

and ECM composition and organisation of both healthy 

and cirrhotic human liver tissue. This is a key advance in the 

development of 3D-technologies for the study of the progression 

of human liver fibrosis into cirrhosis and hepatocellular 

carcinoma. 

Disclosures: 




Gilead, BMS 

The following authors have nothing to disclose: Giuseppe Mazza, Lisa Longato, 

Walid Al-Akkad, Andrea Telese, Luca Urbani, Andrew R. Hall, Benjamin Robinson, 

Luca Frenguelli, Oliver Willacy, Marco Curti, Domenico Tamburrino, Gabriele 

Spoletini, Massimo Malago, Tu Vinh Luong, Armando E. Del Rio Hernandez, 

Paolo De Coppi, Krista Rombouts 

1389 

Rev-erb and TGF-β Differentially Regulate Autophagy in 

Hepatic Stellate Cells 

Paul G. Thomes 1 , Nicole A. Feilen 1 , Jennifer H. Benbow 1 , Elizabeth 

Brandon-Warner 1 , Cathy Culberson 1 , Terrence M. Donohue 2 , 

Laura W. Schrum 1 ; 1 Liver Pathobiology Laboratory, Carolinas 

Medical Center, Charlotte, NC; 2 Omaha VA Medical Center & 

Univ. of Nebraska Medical Center, Omaha, NE 

Background: Recently, we demonstrated that ligand-activated 

nuclear receptor Rev-erbα mitgates the fibrogenic phenotype of 

hepatic stellate cells (HSCs) (Li et al., Hepatology, 2014). Reverbα 

is also a novel regulator of macroautophagy (autophagy) 

(Woldt et al., Nat Med, 2013), a crucial lysosomal degrading 

system that likely provides substrates for HSC activation. Here, 

we examined whether pharmacological activation of Rev-erb 

with SR9009 or treatment with the pro-fibrotic cytokine, TGFβ, 

each differentially modulates autophagy in activated rat 

primary HSCs and in HSC or fibroblast cell lines. Methods: 

We treated activated human HSC cell lines, LX2 and TWNT-4, 

primary rat hepatic stellate cells (rHSCs) or mouse embryonic 

fibroblast cells (3T3) with TGF-β (5 ng/ml) or Rev-erb ligand, 

SR9009 (10 mM), for 24 hours. Autophagy was quantified 

by protein markers and autophagosome (AV) flux by Western 

blot analyses (WB) and immunohistochemistry (IHC). Results: 

IHC demonstrated decreased AV following TGF-β exposure 

in all cell types. Rev-erb activation with SR9009 decreased 

AV in TWNT-4, 3T3 and rHSCs but not in LX2 cells. WBs 

confirmed lower levels of AV marker protein, LC3II, in all TGFβ- 

treated cells even after 4hr of cytokine exposure. In LX2, 

TWNT-4 and rHSCs, TGF-β simultaneously decreased levels of 

P62, an adaptor protein whose levels reflect the degree of AV 

degradation. SR9009-treated LX2, TWNT-4 and 3T3 cells had 

decreased LC3II protein levels and an insignificant decrease 

in rHSCs. P62 levels were unaffected by SR9009 exposure 

in all cell types. TGF-β treatment of rHSC was associated with 

higher lysosome (Lys) (LAMP1) numbers and a higher co-localization 

frequency of AVs with Lys. In contrast, SR9009 treatment 

caused reduced Lys and AV-Lys co-localization. In rHSCs, 

SR9009 but not TGF-β, decreased lysosomal cathepsin B activity. 

When cells were co-treated with TGF-β and bafilomycin 

(blocks AV degradation), rHSCs exhibited numerically higher 

LC3II levels than cells treated with bafilomycin alone. Rat HSCs 

treated with SR9009 and bafilomycin exhibited significantly 

lower LC3II levels than cells treated with bafilomycin alone. 

In rHSC, TGF-β and the autophagy inducers, rapamycin and 

starvation decreased Rev-erbα nuclear staining. SR9009 treatment 

decreased Rev-erbα cytoplasmic staining. Conclusion: 

Our findings indicate that TGF-β accelerates AV degradation 

and SR9009 retards AV synthesis and degradation in activated 

HSCs. We propose that Rev-erbα is a transcriptional regulator 

of autophagy in HSCs and that its ligand, SR9009, slows autophagic 

degradation of the macromolecular substrates that fuel 

fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Paul G. Thomes, Nicole A. Feilen, 

Jennifer H. Benbow, Elizabeth Brandon-Warner, Cathy Culberson, Terrence M. 

Donohue, Laura W. Schrum 

1390 

Molecular mechanism responsible for tissue stiffness in 

advanced chronic liver fibrogenesis 

Takao Sakai; Department of Molecular and Clinical Pharmacology, 

Institute of Translational Medicine, University of Liverpool, 

Liverpool, United Kingdom 

Tissue fibrosis is a part of wound-healing response that maintains 

organ structure and integrity following tissue damage 

and characterized by extracellular matrix (ECM) remodeling 

and stiffening. However, functional contribution of tissue stiffening 

to non-cancer pathogenesis remains largely unknown. In 

particular, how remodeling of ECM by myofibroblasts results 

in changes in mechanical tension that support the activation 

of pathogenic signaling pathways remain to be elucidated. 

Fibronectin is an ECM glycoprotein substantially expressed 

during adult tissue repair. We have addressed the molecular 

mechanism responsible for tissue stiffness in advanced chronic 

liver fibrogenesis induced by carbon tetrachloride (16 weeks) 

using a mouse model lacking fibronectin in the adult liver. 

Fibronectin-null livers exhibited constitutively elevated local 

TGF-β activity, induced more myofibroblast phenotypes, and 

accumulated highly disorganized/diffuse collagenous ECM 

networks composed of thinner and significantly increased number 

of collagen fibrils during advanced chronic liver damage. 

Consequently, fibronectin-null livers lead to more extensive liver


cirrhosis, which was accompanied by significantly increased 

liver tissue stiffness and deteriorated hepatic functions. Mechanistically, 

mutant livers showed elevated lysyl oxidase expressions, 

and a significant amount of active lysyl oxidase was 

released in fibronectin-null hepatic stellate cells in response 

to TGF-β1 through both canonical Smad- and non-canonical 

such as PI3 kinase-mediated pathways. Furthermore, inhibition 

of lysyl oxidase resulted in the disruption of TGF-β-induced 

fine collagen fibril networks and significantly reduced 

formed collagen fibril stiffness, and treatment of fibronectin in 

fibronectin-null stellate cells recovered collagen fibril stiffness to 

wild-type levels. Thus, our novel findings demonstrate an indispensable 

role for fibronectin in chronic liver fibrosis/cirrhosis 

in negatively regulating TGF-β bioavailability, which in turn 

modulates ECM remodeling and stiffening, and consequently 

preserves adult tissue functions. 

Disclosures: 

The following authors have nothing to disclose: Takao Sakai 

1391 

Antifibrogenic properties of monoacylglycerol lipase 

inhibitors 

Aida Habib 1 , Jinghong Wan 1 , Pushpa Hegde 1 , Emmanuel Weiss 1 , 

Arthur Brouillet 2 , Richard Moreau 1 , Sophie Lotersztajn 1 ; 1 Inserm 

U1149, Center for Research on inflammation, Paris, France; 

2 Inserm U955, Paris, France 

Background and aims Monoacylglycerol lipase (MAGL) is the 

rate-limiting enzyme in the degradation of monoacylglycerols. 

In addition to its role in lipid metabolism, MAGL is a pivotal 

component of the endocannabinoid system, since this enzyme 

metabolizes 2-arachydonoyl-glycerol, an endogenous cannabinoid 

receptor ligand, into arachidonic acid. Recent studies 

have identified inhibition of MAGL as an interesting anti-inflammatory 

strategy in several experimental models of chronic 

inflammatory diseases. In the present study, we investigated 

the impact of MAGL inhibition on inflammation and fibrosis 

regression. Methods: Liver fibrosis regression was studied in 

C57BL/6 mice exposed to chronic administration of carbon 

tertrachloride (CCl 4 

) for 6 weeks. Injections were discontinued 

and mice were daily administered with the MAGL inhibitor 

JZL184 (15 mg/k, i.p.) or vehicle for up to 4 days. In vitro 

studies were performed on isolated murine peritoneal macrophages 

and hepatic myofibroblasts, and on PBMC from alcoholic 

cirrhotic and healthy subjects. Sirius red staining was 

quantified by morphometric analysis, fibrogenic and inflammatory 

gene expression by RT-PCR analysis, and the identity 

of intrahepatic leucocytes was analyzed by FACS. Results: 

JZL184 accelerated fibrosis regression 4 days after cessation 

of CCl 4 

administration, as shown by a reduction of sirius red 

staining and smooth muscle alpha actin immunolabeling. JZL 

184 also decreased the expression of fibrogenic genes in 

treated vs untreated animals. Moreover, JZL184 significantly 

decreased the influx of Ly6C + infiltrating monocytes into the 

liver of CCl4-exposed mice. Accordingly, JZL 184 reduced the 

hepatic expression of Ly6C mRNA and decreased the hepatic 

mRNA expression of inflammatory genes including IL1-b, CCl2, 

and CCL4. In vitro studies demonstrated that JZL184 reduced 

hepatic myofibroblast proliferation and down-regulated 

LPS-stimulation of inflammatory gene expression in peritoneal 

macrophages. Finally, MAGL mRNA expression was increased 

in PBMC from patients with alcoholic cirrhosis as compared to 

healthy subjects and further enhanced in LPS-exposed PBMC. 

Conclusions: Pharmacological inhibition of MAGL accelerates 

liver fibrosis regression in a model of toxin-induced liver injury, 

most probably by a mechanism involving reduction of infiltrating 

pro-inflammatory monocytes into the liver and a decrease 

in inflammatory mediators produced by macrophages. These 

results unravel MAGL inhibition as a novel promising antifibrogenic 

approach. 

Disclosures: 

The following authors have nothing to disclose: Aida Habib, Jinghong Wan, 

Pushpa Hegde, Emmanuel Weiss, Arthur Brouillet, Richard Moreau, Sophie Lotersztajn 

1392 

Induction of XBP1 Leads to Hepatic Stellate Cell Activation 

Rosa S. Kim, Daisuke Hasegawa, Xiaochen Sun, Takuma 

Tsuchida, Dipankar Bhattacharya, Hsin I Chou, David Y. Zhang, 

Yujin Hoshida, Scott L. Friedman; Liver Diseases, Icahn School of 

Medicine at Mount Sinai, New York, NY 

BACKGROUND AND AIM: We have previously established 

that ER stress leading to the Unfolded Protein Response (UPR) 

promotes hepatic stellate cell (HSC) activation in liver injury 

(Hernandez-Gea, V., et al., J Hepatol, 2013. and Hasegawa, 

D., et al., PLoS ONE 2010.). UPR has three effector pathways, 

PERK, ATF6 and IRE1-XBP1. IRE1 splices uXBP1 into its active 

transcriptional effector, sXBP1. Our aim was to define the 

relative contribution of XBP1 to HSC activation. METHODS: 

We overexpressed XBP1 splice isoforms 

by lentiviral infection for 

96 hours in primary human HSCs and immortalized human 

HSC lines (TWNT4, LX2). Fibrogenic targets were assessed by 

qPCR and immunoblot (collagen I, αSMA, βPDGFR, MMP2, 

TIMP1). Genome-wide transcriptome profiling was performed 

using HumanHT-12 beadarrays (Illumina) to define changes in 

global gene expression and pathway engagement downstream 

of XBP1 splice isoforms 

. Stringent criteria (FDR


1393 

Role of CREBH activation induced by HCV infection in 

up-regulation of TGF-β2 expression and fibrogenesis 

Takeshi Chida 1,2 , Masahiko Ito 1 , Kazuhito Kawata 2 , Yoshimasa 

Kobayashi 2 , Tetsuro Suzuki 1 ; 1 Infectious disease, Hamamatsu university 

school of medicine, Hamamatsu, Japan; 2 2nd department 

of internal medicine, Hamamatsu university school of medicine, 

Hamamatsu, Japan 

Background and Aim: The mechanisms by which infection 

with hepatitis C virus (HCV) modulates the process of liver 

fibrosis still remains poorly defined. Increased expression of 

pro-fibrogenic growth factors including transforming growth 

factor-β (TGF-β) and extra cellular matrix proteins has been 

correlated with the degree of fibrosis in the liver. The aim of 

this study was to elucidate the mechanism of increased expression 

of TGF-β mediated by HCV infection. Methods: Fibrogenic 

responses caused by HCV (J6/JFH-1 isolate) infection was 

assessed using co-culture system. Hepatocellular carcinoma 

Huh7 cells infected with HCV were co-cultured with hepatic stellate 

TWNT4 cells. TGF-β promoter activities were analyzed by 

the luciferase reporter assay. Expression of mRNAs of fibrotic 

markers was analyzed by real-time RT-PCR. Protein expression 

was assessed by Western blotting and immunofluorescence. 

Knockdown and knockout for targeted genes were performed 

using siRNA and CRISPR-Cas9 systems, respectively. Results: 

Expression of TGF-β1 and TGF-β2 was significantly higher 

in HCV-infected cells than in non-infected cells; in particular, 

gene expression and promoter activity of TGF-β2 were markedly 

increased in cells with HCV infection or expression of the 

viral polyproteins such as Core through NS2. Compared to 

HCV-infected Huh7 alone or TWNT4 alone, co-culture of the 

infected Huh7 and TWNT4 cells showed significantly higher 

expression of collagen type1A1 (COL1A1). The upregulation 

of COL1A1 was suppressed by TGF-β2 knockout in Huh7 cells. 

From mutagenesis together with search for transcription factor 

binding sites, two regions; previously-identified CRE and 

temporal “CRE binding protein, hepatocyte-specific (CRE- 

BH)-response” element (CREBHRE) within the proximal TGF-β2 

promoter are important for its transcription regulation. Gel shift 

assay showed CREBH binding to the regions. TGF-β2 promoter 

activity was decreased by CREBH knockdown or knockout, and 

was increased by expression of the active form of CREBH. ChIP 

assay showed that binding of CREBH and ATF2 to TGF-β2 promoter 

was increased when the HCV Core-NS2 was expressed, 

under which precursor CREBH was processed into its active 

form. Discussion: CREBH, an endoplasmic reticulum (ER)-localized, 

liver-enriched transcriptional factor, is well-known to be 

activated by ER-stress. Our findings suggest that the expression 

of HCV proteins in the virus-infected cells potentially induces 

ER-stress and CREBH activation. Increased expression of TGFβ2 

by CREBH activation and its release from the infected cells 

contributes to the induction of fibrogenic responses via paracrine 

signaling to nearby hepatic stellate cells. 

Disclosures: 

The following authors have nothing to disclose: Takeshi Chida, Masahiko Ito, 

Kazuhito Kawata, Yoshimasa Kobayashi, Tetsuro Suzuki 

1394 

Expression of ENTPD1/CD39 is protective in a mouse 

model of biliary fibrosis 

Linda Feldbrügge 1,3 , Shuji Mitsuhashi 1 , Eva Csizmadia 1 , Xiaofeng 

Sun 1 , Moritz Schmelzle 2 , Simon C. Robson 1 ; 1 Gastroenterology, 

BIDMC, Boston, MA; 2 Department for Surgery, Charité, Berlin, 

Germany; 3 Department for Surgery, University Hospital Leipzig, 

Leipzig, Germany 

Introduction. Ecto-nucleoside triphosphate diphosphohydrolases 

(ENTPD) comprise a family of cell surface located 

transmembrane proteins that regulate purinergic signaling by 

catalyzing extracellular nucleotides, such as ATP, ADP, to ultimately 

generate adenosine. These mediators are important signaling 

molecules in hepatic injury and inflammation. ENTPD1/ 

CD39 is expressed on endothelium, and sinusoidal immune 

cells, and closely regulates liver regeneration by generating 

AMP from ATP and ADP. ENTPD2/CD39L1 is expressed on 

portal fibroblasts and perivascular cells and is a preferential 

ecto-ATPase, generating ADP. The role of ENTPD-mediated 

catalysis in modulating liver and biliary fibrosis is currently 

unclear as both ATP and adenosine could promote fibrogenic 

signals in portal myofibroblast and stellate cells and ADP is a 

potent platelet agonist. Methods. C57BL6 wild type, ENTPD1 

null mice and ENTPD2 null mice were subjected to bile duct 

ligation (BDL). Blood and liver tissue were harvested at 2 and 

4 weeks after BDL. Liver function, tissue injury and extent of 

cholestasis were determined by serum liver function tests and 

histopathology. To assess the extent of biliary fibrosis, we also 

analyzed tissue hydroxyproline content and performed Masson’s 

trichrome staining. Results. We observe a trend towards 

decreased survival in the ENTPD1 null mice after BDL. ENTPD1 

null mice further show significantly more weight loss, develop 

more pronounced cholestasis, and more severe liver fibrosis 

than wild type mice. No major profibrogenic effects are noted 

in ENTPD2 null mice, as determined by hydroxyproline content 

and morphometric analysis of peribiliary collagen deposition. 

Conclusions. Purinergic signaling regulated by ENTPD1 limits 

biliary fibrosis. In contrast, ENTPD2 seems to have less pronounced 

effects. These opposing roles are most likely due to 

differential catalytic function and localization of both enzymes 

with ENTPD1 as the dominant, vascular endothelial ecto-enzyme. 

Our findings further suggest a dominant role of ATP-mediated 

purinergic receptor signal transduction during biliary 

fibrogenesis. 

Disclosures: 

Moritz Schmelzle - Grant/Research Support: Novartis GmbH 





The following authors have nothing to disclose: Linda Feldbrügge, Shuji Mitsuhashi, 

Eva Csizmadia, Xiaofeng Sun


1395 

Inhibition of microRNA-214 ameliorates hepatic fibrosis 

and tumor incidence in platelet-derived growth factor 

C transgenic mice by rescuing Mig-6, EGFR/MET signal 

inhibitor. 

Hikari Okada 1 , Masao Honda 1 , Jean S. Campbell 2 , Kai Takegoshi 

1 , Yoshio Sakai 1 , Taro Yamashita 1 , Takayoshi Shirasaki 1 , 

Riuta Takabatake 1 , Mikiko Nakamura 1 , Takuji Tanaka 3 , Shuichi 

Kaneko 1 ; 1 Gastroenterology, Kanazawa University Graduate 

School of Medical Science, Kanazawa, Japan; 2 Department of 

Pathology, University of Washington School of Medicine, Seattle, 

WA; 3 Cancer Research and Prevention, The Tohkai Cytopathology 

Institute, Gifu, Japan 

Objective Overexpression of platelet-derived growth factor-C 

(PDGF-C) in the liver of mice (Pdgf-c Tg) induces hepatic fibrosis, 

followed by the development of hepatocellular carcinoma. 

We identified differentially expressed miRNAs in Pdgf-c Tg 

mice and evaluated their functional relevance in the progression 

of hepatic fibrosis and the development of HCC. Materials 

and Method Pdgf-c Tg mice at 9 weeks and 32 months 

of age were injected with LNA-antimiR-214 via the tail vein 

twice at an interval of 7 days and six times (50 μg each) with 

3-week intervals by using Invivofectamine® 2.0. The degree 

of hepatic fibrosis, tumor incidence, tumor number, and liver 

weight were calculated. The expression of collagens I and IV, 

alpha-smooth muscle actin (α-SMA), Mig-6 (Errfi1), p-SMAD3, 

p-EGFR, p-MET, p-AKT and p-ERK was evaluated by immunohistochemical 

staining, real-time PCR, and Western blotting. To 

determine the target of miR-214, we used human HCC cell lines 

Huh-7 and HLF and immortalized human stellate cells Lx-2 in 

vitro. Results MiR-214 was the most significantly up-regulated 

miRNA in the liver of Pdgf-c Tg mice. Moreover, we showed the 

expression of miR-214 was up-regulated with the progression 

of hepatic fibrosis in a diet-induced (Ath+HF) NASH mouse 

model and in patients with chronic hepatitis B or C. Experiments 

of co-culture of hepatocytes (Huh-7 cells) and stellate cells 

(Lx-2 cells) revealed that miR-214 is transferred to hepatocytes 

by exosomes, which are released from activated stellate cells. 

Pdgf-c Tg mice treated with LNA-antimiR-214 showed a marked 

reduction in fibrosis and tumor incidence compared with salineor 

LNA-miR-control-injected control mice. We found mir-214 

targeted a negative regulator of EGFR signaling, Mig-6. Mimic-miR-214 

decreased the expression of Mig-6 and increased 

the levels of EGF-mediated p-EGFR (Y1173 and Y845) and 

p-Met (Tyr1234/1235), then increased growth of Huh-7 cells. 

Conversely, LNA-antimiR-214 repressed the expression of these 

genes and decreased cell growth. Similarly, Mig-6 could be a 

target of miR-214 in Lx-2 cells. The expression of Mig-6 was 

induced by LNA- antimiR-214 and over expression of Mig-6 

repressed the expression of COLA2, COL4A1, PDGFRβ, p-AKT 

and p-ERK in Lx-2 cells. Conclusion These results demonstrate 

that miR-214 participates in the development of hepatic fibrosis 

and tumor by targeting EGFR/MET signal inhibitor, Mig6. 

LNA-antimiR-214 is therefore potentially useful in the prevention 

of hepatic fibrosis and HCC. 

Disclosures: 


Jean S. Campbell - Employment: OncoSec Medical 






The following authors have nothing to disclose: Masao Honda, Kai Takegoshi, 

Yoshio Sakai, Taro Yamashita, Takayoshi Shirasaki, Riuta Takabatake, Mikiko 

Nakamura, Takuji Tanaka 

1396 

Essential Roles of RNA-binding Protein HuR in Activation 

of Hepatic Stellate Cells Induced by Transforming 

Growth Factor-β1 

Jingjing Ge, Na Chang, Zhongxin Zhao, Lei Tian, Lin Yang, Liying 

Li; Department of Cell Biology, Capital Medical University, Beijing, 

China 

Sphingosine kinase 1 (SphK1) is involved in transforming 

growth factor-β1 (TGF-β1)-induced activation of hepatic stellate 

cells (HSCs), which is characterized by increased expression 

of α-smooth muscle actin (α-SMA) and Collagen α1(I) in 

liver fibrogenesis. RNA-binding protein HuR up-regulates TGFβ1 

expression and mediates TGF-β1-induced profibrogenic 

actions. However, the molecular mechanism by which TGFβ1 

regulates SphK1 remains undefined. Here we investigated 

the role of HuR in SphK1 expression. Mice liver fibrosis were 

induced by bile duct ligation or carbon tetrachloride treatment. 

Expressions of HuR, SphK1, α-SMA and Collagen α1(I) in the 

fibrotic liver or human HSCs cell line (LX-2) were measured by 

real-time RT-PCR or Western blot. RNA immunoprecipitation 

was performed to detect the association between HuR and 

SphK1 mRNA. We found that HuR mRNA was increased in 

the damaged liver and had positive correlation with mRNA 

of TGF-β1, SphK1, α-SMA and Collagen α1(I), respectively. 

Importantly, up-regulation of SphK1 and activation of HSCs 

stimulated by TGF-β1 depended on HuR cytoplasmic accumulation, 

as HuR cytoplasmic translocation was blocked by Leptomycin 

B (LMB, a specific nuclear export inhibitor), the effects 

of TGF-β1 were diminished. In addition, silencing HuR significantly 

abolished the up-regulation of SphK1 and activation of 

HSCs evoked by TGF-β1, and overexpressing HuR mimicked 

the effects of TGF-β1. Furthermore, TGF-β1 prolonged half-life 

of SphK1 mRNA by promoting its binding with HuR. Pharmacological 

or siRNA-induced inhibition of SphK1 abrogated 

HuR-mediated activation of HSCs. In conclusion, these data 

suggest that HuR binds with SphK1 mRNA and plays a crucial 

role in TGF-β1-induced activation of HSCs. 

Disclosures: 

The following authors have nothing to disclose: Jingjing Ge, Na Chang, Zhongxin 

Zhao, Lei Tian, Lin Yang, Liying Li 

1397 

HIV infected Kupffer cells mediate pro-fibrogenic effects 

on stellate cells through secretion of TGFβ1 

Ankur Panchal, Arevik Mosoian, Lumin Zhang, M. Isabel Fiel, 

Sander S. Florman, Myron E. Schwartz, Andrea D. Branch, Meena 

B. Bansal; Icahn School of Medicine at Mount Sinai, New York, 

NY 

Background: Liver disease is the most common cause of 

non-AIDS related mortality in HIV-infected patients receiving 

effective anti-retroviral therapies. TGFβ1 is an important profibrogenic 

cytokine in chronic liver disease and both serum 

and intrahepatic levels of TGFβ1 are increased in both HIV 

monoinfected and HIV/HCV coinfected patients, respectively. 

As the resident liver macrophage, the Kupffer cell, has been 

shown to be susceptible to HIV infection in vitro and in vivo 

and is the predominant cellular source of intrahepatic TGFβ1, 

we hypothesized that HIV infection of Kupffer cells (KCs) results 

in increased TGFβ1 secretion with subsequent paracrine pro-fibrogenic 

effects on hepatic stellate cells (HSCs), the primary 

collagen producing cell in liver fibrosis. Methods: Primary KCs 

and HSCs were isolated by density centrifugation from normal 

livers in patients undergoing hepatic resection for primary 

benign tumors or a single metastasis from colon cancer. KC


preps were enriched by rapid adherence. Purity of KCs was 

assessed by RT-PCR and FACS for CD68, CD14, TLR4, CD3, 

and CD31. Primary KCs were infected with HIV-BaL (MOI=0.1) 

or mock infected and supernatants were collected at serial 

time points for p24, a marker of HIV replication, and TGFβ1 

ELISA. Pro-fibrogenic effects of conditioned media from KCs on 

HSCs was assessed by qRT-PCR and specific effects of TGFβ1 

were assessed using TGFβ1 blocking/neutralizing antibody. 

Recombinant TGFβ1 (10ng/mL) was used as a positive control. 

Results: 1) HIV Infection of KCs resulted in an 1.5 fold increase 

in TGFβ1 secretion (p


sustained to be increased in TAA+Vector group at week 3 

whereas TAA+Relaxin rats had suppressed TIMP-2 expression 

by then. Liver α-smooth muscle actin (α-SMA) mRNA in 

both TAA+Vector and TAA+Relaxin rats was increased after 

3 days but reversed to the normal level at week 3. Analysis 

of TGFβ and MMP9 mRNA demonstrated the similar pattern. 

Conclusions: A single adenoviral delivery of relaxin in the liver 

attenuated established hepatic fibrosis by suppressing collagen 

cross-links and enhancing collagen degradation. 

Disclosures: 

The following authors have nothing to disclose: Jung Il Lee, Ja Kyung Kim, Hye 

Young Chang, Kwan Sik Lee 

1400 

Galectin-1 promotes the activation of hepatic stellate 

cell and liver fibrosis through glycosylation-dependent 

interactions 

Ming-Heng Wu; Graduate Institute of Translational Medicine, College 

of Medical Sciences and Technology, Taipei Medical University, 


The activation of hepatic stellate cell (HSC) is a rate-limiting 

step in hepatic fibrosis. Therefore, developing approaches 

to destroy or normalize activated HSCs is important for liver 

fibrosis therapy. The coevolution of cell-surface glycans and 

glycan-binding proteins regulate the progression of physiologic 

and pathologic processes but this issue has not been 

well addressed in HSC activation and liver fibrosis. Here, we 

reported that galectin-1 (Gal-1), a β-galactoside binding protein, 

was highly expressed in the fibrotic livers. Gal-1 binding 

glycans [β1,6 N-acetylglucosamine-branched N-glycans and 

poly-N-acetylactosamine (LacNAc)] were highly presented in 

activated HSCs and facilitated Gal-1 binding. In contrast, the 

amounts of α2,6 sialylation glycan (which masks the Gal1 

binding glycan) is lower in activated HSCs than in quiescent 

HSCs. Two glycosyltransferases, Mgat5 (N-acetylglucosaminyltransferase 

5) and GCNT1 (core 2 N-acetylglucosaminyltransferase 

1), mediated N- and O-glycans were required for 

Gal-1 stimulated migration and activation of HSCs. In addition, 

Gal-1 regulated PDGF and TGF-β1 induced signaling and 

HSC activation through interacting with neuropilin-1 (NRP-1). 

Using a mouse model, we found that thioacetamide (TAA), an 

organosulfur compounds, induced liver fibrosis was attenuated 

in Gal-1 null mice. In summary, our results demonstrated that 

the crucial role of Gal-1 in HSC activation and liver fibrosis. 

Strategies that disrupt the Gal-1/NRP-1 interactions could be 

developed for liver fibrosis therapy. 

Disclosures: 

The following authors have nothing to disclose: Ming-Heng Wu 

1401 

TRPC6 Channels Contribute to Hepatic Stellate Cell Activation 

Leading to Liver Fibrosis 

Kyu-Hee Hwang, Ji-Hee Kim, Soo-Jin Kim, Kyu-Sang Park, Seung- 

Kuy Cha; Physiology, Yonsei University Wonju College of Medicine, 

Wonju, Korea (the Republic of) 

Activation of hepatic stellate cells (HSCs) is a main cause of 

liver cirrhosis and portal hypertension. In response to injury, 

HSCs are activated by diverse hormones and growth factors 

such as angiotensin II and endothelin-1 whose receptors are 

linked to phospholipase activation leading to Ca 2+ influx 

termed receptor-operated Ca 2+ entry (ROCE). The Ca 2+ -mediated 

signaling pathways are implicated either directly or 

indirectly in activation of HSCs causing de novo expression of 

a-smooth muscle actin (αSMA) and/or profibrotic ligand TGFβ. 

However, the molecular identity and underlying mechanism of 

ROCE involving HSC activation are ill-defined. Here, we report 

that TRPC6 channel is a predominant molecular component 

of ROCE mediating HSC activation. Among TRPC sub-family, 

the TRPC6 expression was significantly increased in the bile 

duct ligation- and TAA-induced liver fibrosis animal models. 

Functionally, TRPC6-stimulated Ca 2+ influx was confirmed by 

specific blockade of TRPC6 with the siRNA against Trpc6 and 

pharmacological inhibitor SKF96365 in cultured human HSCs. 

Furthermore, overexpression of TRPC6 by gene delivery in 

mouse liver induced de novo expression of αSMA and TGFβ 

suggesting that TRPC6-mediated Ca 2+ influx may involve in 

HSC activation and liver fibrosis. These results provide a new 

perspective on the pathogenesis of liver fibrosis and may provide 

clues to treatment of the liver cirrhosis. 

Disclosures: 

The following authors have nothing to disclose: Kyu-Hee Hwang, Ji-Hee Kim, Soo- 

Jin Kim, Kyu-Sang Park, Seung-Kuy Cha 

1402 

Targeting Activated Stromal Fibroblasts in Primary Sclerosing 

Cholangitis 

Anja Moncsek 1 , Achim Weber 2 , Beat Müllhaupt 1 , Joachim C. 

Mertens 1 ; 1 Gastroenterology and Hepatology, University Hospital 

Zürich, Zürich, Switzerland; 2 Surgical Pathology, University Hospital 

Zürich, Zürich, Switzerland 

Background: Primary Sclerosing Cholangitis (PSC) is a chronic 

cholestatic condition with diffuse biliary inflammation and fibrosis 

that can affect the whole biliary system. The disease has 

a progressive course and results in biliary cirrhosis with all 

its sequelae. PSC is also one of the main known risk factors 

for the development of cholangiocarcinoma (CCA). We have 

previously shown that activated stromal fibroblasts (ASF) in 

cholangiocarcinoma, which have been implicated in tumor 

progression and metastasis display an activated phenotype 

with increased sensitivity to apoptotic stimuli and can be selectively 

targeted with pro-apoptotic BH3 mimetics (Mertens et.al. 

Cancer Res 2013). However, the role of ASF in PSC and progression 

to CCA is not well understood. Thus, our aim was 

to characterize ASF in PSC and explore selective pro-apoptotic 

targeting of this cell population. Methods: Human PSC 

specimens were examined by histology, immunofluorescence, 

Western blot and qRT-PCR. The MDR2 -/- mouse model was 

employed for in vivo studies. Results: The PSC tissue specimens 

we examined contain an abundance of activated stromal fibroblasts. 

By immunofluorescence the stromal compartment stains 

positive for Tenascin C and Collagen I, while the stromal fibroblasts 

are positive for alpha smooth muscle actin (aSMA). This 

phenotype resembles ASF in CCA. We were able to confirm 

the phenotype of PSC-ASF by PCR. The MDR2 -/- mouse model 

of PSC develops a periductal and bridging fibrosis within 8 to 

12 weeks. In analogy to previous experiments in a rat model 

of CCA, MDR2 -/- mice were treated with the pro-apoptotic BH3 

mimetic navitoclax for 2 weeks. Assessment of liver fibrosis by 

Hydroxyproline assay and Sirius red staining demonstrated a 

reduction in fibrosis. Preliminary results revealed a reduction of 

activated stromal fibroblasts. Conclusions: ASF in PSC resemble 

stromal fibroblasts in cholangiocarcinoma and display apoptotic 

sensitization that could be targeted therapeutically. 

Disclosures: 

The following authors have nothing to disclose: Anja Moncsek, Achim Weber, 

Beat Müllhaupt, Joachim C. Mertens


1403 

Modulation of Extracellular Matrix Remodeling by Activation 

of the Aryl Hydrocarbon Receptor 

Cheri L. Lamb 1,2 , Kristen A. Mitchell 1,2 ; 1 Biological Sciences, Boise 

State University, Boise, ID; 2 Biomolecular Sciences Ph.D. Program, 

Boise State University, Boise, ID 

The aryl hydrocarbon receptor (AhR) is a soluble, ligand-activated 

transcription factor involved in developmental processes, 

xenobiotic metabolism, and adaptation to environmental stress. 

Recent evidence indicates that this receptor may also contribute 

to wound healing processes. We recently found that exposure 

to the potent AhR agonist 2,3,7,8 tetrachlorodibenzo-p-dioxin 

(TCDD) increases activation of hepatic stellate cells, which 

contribute to hepatic wound healing and fibrosis through the 

deposition of extracellular matrix material, namely collagen 

type I. The goal of the present study was to determine how 

TCDD treatment impacts matrix deposition and remodeling 

activities during experimental liver fibrosis elicited by carbon 

tetrachloride (CCl 4 

). Male C57Bl/6 mice were treated with 

CCl 4 

(5 ml/kg diluted 1:10 in corn oil) twice a week for eight 

weeks. During the final two weeks, mice were treated with 

TCDD (20 μg/kg by gavage) or vehicle (peanut oil). Collagen 

deposition was measured by cytohistological staining and 

evaluated by polarized microscopy. Levels of matrix enzymes 

and associated regulatory proteins were measured by qPCR, 

and activity was assessed by in situ zymography. Results indicate 

that TCDD treatment increased Col1a1 mRNA levels and 

altered the pattern of collagen deposition in the fibrotic septa. 

Exposure to TCDD also increased in situ collagenase activity 

in the fibrotic liver with a concomitant 4-fold increase in matrix 

metalloproteinase-13 mRNA levels. Inhibitors of matrix metalloproteinases, 

namely PAI-1 and TIMP-1, were increased in 

TCDD-treated mice, which could reflect direct AhR transcriptional 

activity or else a compensatory response to elevated 

collagenase activity. Collectively, these results indicate that 

TCDD-induced AhR activation not only increases fibrogenesis, 

but also perturbs extracellular matrix remodeling activities in 

the fibrotic liver. Hence, these findings implicate a previously 

unidentified role for the AhR in modulating wound healing 

responses in the liver and may provide a basis for designing 

therapeutic strategies to limit fibrosis by modulating AhR activation. 

Disclosures: 

The following authors have nothing to disclose: Cheri L. Lamb, Kristen A. Mitchell 

1404 

MicroRNA profiling of circulating exosomes in HBV cirrhosis 

patients following anti-HBV therapy 

Min Cong 1 , Li Chen 2 , Tianhui Liu 1 , Ping Wang 1 , Jidong Jia 1 , David 

Brigstock 2 , Hong You 1 ; 1 Liver Research Center, Beijing Friendship 

Hospital, Beijing, China; 2 The research Institute at Nationwide 

Children’s Hospital, Columbus, OH 

Aim: The objective of our study is to establish serum exosomal 

microRNA (miR) signatures to predict progression and reversion 

of fibrosis in hepatitis B (HBV)–induced cirrhosis undergoing 

anti-viral therapy. Methods: Twelve HBV patients with 

compensated cirrhosis were selected based on their cirrhosis 

regression (decreased Fibroscan values of > 5kPa) or progression 

(increased or stable Fibroscan values) after treatment for 6 

months with Entecavir (0.5mg/day oral), respectively. Twelve 

matched healthy blood donors were used as a reference control 

group. Serum (0.5ml) from the HBV patients, before or 

after therapy, or from the controls was individually processed 

to purify exosomes from which small RNA was then isolated. 

Samples in each group were pooled and analyzed using a 

Human miRNome PCR array to profile the1066 most abundantly 

expressed human miR sequences using SYBR Green real 

time PCR. Circulating exosomalmiRs were isolated from 8 other 

compensated cirrhosis HBV patients who presented with cirrhosis 

regression or progression to validate the change of selected 

exosomalmiRs by qRT-PCR. Results: Compared with control 

subjects, 59 out of 1066 miRs from 12 regressed cirrhosis 

HBV patients were up- or down-regulated at least 15-fold in the 

cirrhotic patients, with 46 out of 59 miRs showing correction 

(to more healthy) after therapy. Among the miRs identified in 

this unbiased analysis were miR-7, -15b, -16, -22, -26a, -27b, 

-29, -92b, -101, -183, -191, -196a, -200a, -370, -378a and 

-936 and let-7a-g which have been reported in the literature to 

be fibrosis-related in the liver or other organ systems. Analysis 

of exosomalmiRs from 12 progressed cirrhosis HBV patients 

showed that these miRs were less corrected as compared to 

the regressed patients and that different miRs served to discriminate 

these patients from the healthy controls. qRT-PCR analysis 

of exosomalmiRs from 8 cirrhosis HBV patients, before or after 

therapy, showed that miR-16,-20a,-25,-148a,-223,-320d were 

significantly decreased while miR-126,-191 were significantly 

increased in progressed patients. Conclusions: These data 

reveal that the circulating exosomalmiR content varies according 

to the stage of HBV fibrosis and that exosomalmiRs show 

quantitative and qualitative changes that reflect the patients’ 

clinical course of fibrosis regression or progression after anti-viral 

treatment. Since circulating exosomalmiRs change dynamically 

with disease status, these findings highlight the possibility 

of identifying discriminatory miR signatures that are diagnostic 

or predictive for fibrosis and that can be obtained longitudinally 

and non-invasively in individual patients. 

Disclosures: 

Jidong Jia - Consulting: BMS, MSD, Roche 


The following authors have nothing to disclose: Min Cong, Li Chen, Tianhui Liu, 

Ping Wang, Hong You 

1405 

Identification of Axon guidance signaling pathway 

mediators in hepatic stellate cells and liver fibrogenesis 

Jinsheng Guo 1 , David Zhang 2 , Yujing Wu 1 , Daisuke Hasegawa 2 , 

Scott L. Friedman 1,2 ; 1 Division of Digestive Diseases, Zhong Shan 

Hospital, Fu Dan University, Shanghai, China; 2 Division of Liver 

Diseases, Icahn School of Medicine at Mount Sinai, New York, NY 

Background&Aims: Hepatic stellate cells (HSC) have many 

features of neural cells including expression of glial fibrillary 

protein, nestin and neurotrophin receptors. We recently used 

an informatics based approach to identify novel transcripts 

expressed by stellate cells encoding cell surface and secreted 

proteins (Zhang DY et al, Gut, 2015, in press). Among these, 

Robo2 is a cell surface molecule implicated in axon guidance. 

The aim of our study was to identify the novel signaling pathway 

and key molecules in liver fibrogenesis by dynamic network 

analysis of liver transcriptomes. Methods&Results: Two 

experimental models of hepatic fibrosis were employed by 

intraperitoneally injection of diethylnitrosamine (DEN, 10mg/ 

kg/week) in mice for 14 weeks, or by intraperitoneal injections 

of carbon tetrachloride (CCl4, 0.9ml/kg/biw) for four weeks. 

In the DMN model, the liver samples were collected at 0, 6, 8 

and 14 weeks post DEN treatment for RNA sequencing. The 

transcriptomic data associated with development of liver fibrosis 

(i.e., injury, early fibrotic, late fibrotic or cirrhotic stages) 

were applied to time-series analysis, followed by network 

and pathway analyses. Among markedly up-regulated genes


during fibrosis and cirrhosis, 48 were components of axon 

guidance signaling pathways (P=4.65E-07, Enrichment=2.53), 

which transcripts encoding ligands (slit2, Sema4D), membrane 

receptors (Robo1 and 2, EphA and B, CXCR4, Plexin A4), and 

downstream kinases (PAK1, 2,3 and 6, Gnai 1 and 2, Rock2, 

and Mapk3). By immunoflurescence in immortalized human 

and mouse hepatic stellate cell (HSC) lines LX-2 and JS1 in 

vitro, and in the CCl4 induced fibrotic liver samples in vivo, 

Robo2, a key receptor mediating axon guidance signaling, 

was localized to the HSC surface. The amount and localization 

of its protein expression was correlated well with fibrotic septa 

in fibrotic livers. Conclusion: There is significant upregulation 

and activation of axon guidance signaling pathway members 

in liver during fibrogenesis. This signaling may have regulate 

actin cytoskeleton, cell attraction and outgrowth of HSC. The 

study also revealed RoBo2 as a novel HSC surface marker and 

a potential therapeutic and diagnostic target of liver fibrosis. 

Disclosures: 











The following authors have nothing to disclose: Jinsheng Guo, David Zhang, 

Yujing Wu, Daisuke Hasegawa 

1406 

PDGFRα ubiquitination leads to its p62 mediated autophagic 

degradation in a synectin dependent manner in 

HSC 

Haibin Yu, Mary Drinane, Usman Yaqoob, Vikas K. Verma, Thiago 

de Assuncao, Sheng Cao, Vijay Shah; Gastroenterology Research 

Unit, Mayo Clinic, Rochester, MN 

Background/Aim: The platelet derived growth factor receptor 

(PDGFR) signaling pathway is critical for hepatic stellate cell 

(HSC) activation and liver fibrosis. While the critical role of 

PDGFRβ is well studied, recent work suggests importance of 

PDGFRα as well, making its regulation of interest. Synectin is a 

scaffold protein that is a key regulator of endocytic trafficking 

of multiple tyrosine kinases. Prior studies showed that knockdown 

of synectin from HSC led to autophagic degradation 

of PDGFRα. p62 is a ubiquitin-binding autophagy receptor 

which selectively recognizes autophagic cargo and mediates 

engulfment into autophagosomes in coordination with LC3B. 

We hypothesized that synectin regulates selective autophagic 

degradation of PDGFRα by recognizing specific ubiquitin sites 

which in turn govern p62 binding with PDGFRα. Methods/ 

Results: Co-immunoprecipitation (co-IP) of PDGFRα and PDG- 

FRβ was used to investigate p62 binding using HSC lysates. 

p62 co-precipitated with PDGFRα, but not PDGFRβ. In response 

to PDGF ligand, co-precipitation of PDGFRα with p62 was further 

increased by 4.4 fold (p < 0.05). Synectin knockdown led 

to a 2-fold increase in binding of p62 with PDGFRα (p < 0.05) 

and increased co-localization of p62 and PDGFRα by confocal 

microscopy. Sequence analysis of PDGFRα and PDGFRβ was 

performed which identified two ubiquitin sites present within 

α but not β subunits. Site directed mutagenesis of PDGFRβ 

was performed to mutate the two corresponding residues to 

lysine (Q613K and R979K) and generate a mutant PDGFRβ 

construct (PDGFRβ- Q613K, R979K) that could be used to test 

the hypothesis that these two lysine residues may be critical for 

ubiquitin dependent PDGFR autophagic degradation. PDGFRβ- 

Q613K, R979K underwent autophagic degradation, akin to 

PDGFRα, while the wild-type PDGFRβ did not (p < 0.05). Conclusion: 

The specificity of interaction of the autophagic receptor 

p62 with PDGFR is conferred by specific ubiquitin sites and is 

mediated by synectin. 

Disclosures: 

The following authors have nothing to disclose: Haibin Yu, Mary Drinane, Usman 

Yaqoob, Vikas K. Verma, Thiago de Assuncao, Sheng Cao, Vijay Shah 

1407 

Reactive gamma-ketoaldehydes induce a proinflammatory 

phenotype, oxidative stress, and activation of autophagy 

in primary human hepatic stellate cells 

Lisa Longato 1 , Fausto Andreola 1 , Sean S. Davies 2 , Jackson L. Roberts 

2 , Giuseppe Fusai 1 , Massimo Pinzani 1 , Kevin Moore 1 , Krista 

Rombouts 1 ; 1 Medicine, University College London, London, United 

Kingdom; 2 Vanderbilt University, Nashville, TN 

Background and aims. Products of oxidative stress such as 

4-hydroxynonenal are key activators of hepatic stellate cells 

(HSCs). γ-Ketoaldehydes (γ-KAs), including levuglandins and 

their isomers, are a family of acyclic aldehydes, which are 

formed during oxidation of arachidonic acid, or as a by-product 

of the cyclooxygenase pathway. γ-KAs are highly reactive 

and form protein adducts and cross-links at a rate > 100-fold 

compared to 4-HNE. Increased serum antibodies against proteins 

cross-linked to γ-KAs are present in patients with alcoholic 

liver disease. Since the contribution of γ-KAs to liver injury has 

not been studied, we sought to investigate the effects of γ-KA 

levuglandin E 2 

(LGE 2 

) on the cell biology of primary human 

HSCs. Methods. Primary human HSCs were exposed to LGE 2 

(0.5 pM- 5 mM) for up to 48 hours and analyzed for proliferation, 

cytotoxicity (lactate dehydrogenase, MTS, and Neutral 

Red assays), RNA/protein expression, reactive oxygen 

species (ROS) production, and ultrastructure. Results. Exposure 

to a 5 μM dose of LGE 2 

was profoundly cytotoxic and 

resulted in apoptotic cell death, as indicated by LDH leakage, 

reduced MTS and Neutral Red incorporation, increased levels 

of cleaved PARP, CHOP, and JNK phosphorylation. Lower, 

non-cytotoxic doses (50 nM-500 nM) of LGE 2 

induced selected 

indices of HSCs activation, such as increased expression of 

a-smooth muscle actin (α-SMA), and activation of signaling 

pathways (ERK1/2). In contrast, LGE 2 

had no effect on DNA 

synthesis or pro-fibrogenic markers, particularly collagen type 

I, and lysyl oxidase (LOX). In contrast, HSCs exposed to LGE 2 

displayed a marked increase in expression for various cytokines/chemokines 

including interleukin-8, -6, -1β, and MCP-1. 

This was accompanied by increased secretion of bioactive 

IL1β/IL-18, as measured by HEK-Blue reporter cells. Pre-incubation 

of cells with the either chemicals inhibitors of JNK 

(SP600125), or NFkB (PDTC), but not p38MAPK (SB203580), 

partially reduced these increases in cytokines and chemokines. 

Ultrastructural analyses revealed that LGE 2 

-treated HSCs had 

prominent formation of autophagic vescicles, suggesting ongoing 

autophagy, which was confirmed by immunofluorescence 

for LC3B, as well as by accumulation of LC3II. Lastly, shortterm 

exposure to LGE 2 

promoted a dose-dependent formation 

of ROS in the cells, as measured by the carboxy-H 2 

DCFDA 

fluorescent probe. Conclusions. γ-KAs represent a newly identified 

class of activators of HSCs in vitro, which are biologically 

active at concentrations as low as 50 nM, and are particularly 

effective at promoting a pro-inflammatory response, as well as 

oxidative stress, and autophagy. 

Disclosures: 



Gilead, BMS



The following authors have nothing to disclose: Lisa Longato, Fausto Andreola, 

Sean S. Davies, Jackson L. Roberts, Giuseppe Fusai, Krista Rombouts 

1408 

Sequence Profiling of miRNAs Reveals mir-150-5p as a 

Regulator of Transdifferentiation 

Luigi Locatelli, Pier P. Paoli, Timothy Hardy, Rachel M. Howarth, 

Fiona Oakley, Derek Mann; Institute of Cellular Medicine, Newcastle 

University, Newcastle upon Tyne, United Kingdom 

Cellular transdifferentiation is important in many physiological 

processes (e.g. wound healing), and is implicated in pathologies 

such as cancer and tissue fibrosis. To-date the role of 

miRNAs, a class of small noncoding RNAs that regulate gene 

expression by binding to target mRNAs suppressing its translation 

or initiating its degradation, in transdifferentiation has not 

been fully addressed. We use a cell culture model for transdifferentiation 

of human and rat hepatic stellate cells (HSC) into 

myofibroblasts to determine functional changes in miRNAs. The 

aims of this study were: a) perform Next Generation Sequencing 

(NGS) to generate a miRNA profile in human and rat 

activated HSCs to identify potential miRNA-mRNA regulatory 

networks in common with both; b) replicate NGS results based 

on gene profile and biological validation of specific miRNA 

target genes. The most critical step in miRNA biology lies in the 

definition of the rules for miRNA target recognition. Data generated 

by sequencing were analysed via the Seq-Imp pipeline 

(http://www.ebi.ac.uk/research/enright/software/kraken), to 

identify individual miRNA and an initial statistical analysis. 

Further analysis was performed with a custom R script to generate 

data correlation and heatmap matrices. Finally, targets for 

each miRNA were identified from TargetScan, microCOSM, 

and miRTar, by picking genes that were in at least two out of 

three databases, revealing 11 significantly upregulated and 11 

downregulated miRNAs that were identical in both activated 

human and rat HSC. From the list of differentially regulated 

miRNAs, we focused our attention on mir-150-5p, which regulates 

a series of target proteins such as the Zinc finger E-box 

binding homeobox (ZEB1) involved in the cell cycle, cellular 

development, and apoptosis. Particularly mir-150-5p has been 

found to be downregulated in both human and rat activated 

HSCs. A significant inverse correlation between ZEB1 and mir- 

150-5p was found at the RNA and protein level. In vitro we 

showed that the overexpression of mir-150-5p in human and 

rat HSCs by a mir-150-5p mimic was able to downregulate 

the expression of ZEB1 with an observed reduction of extracellular 

matrix proteins collagen type I (COL1A1) and α-smooth 

muscle actin (α-SMA). Moreover, the level of mir-150-5p in the 

serum of 10 Alcoholic Liver Disease (ALD) patients has been 

found to be significantly downregulated compared to healthy 

controls. In conclusion, HSC transdifferentiation is associated 

with discrete alterations in the expression of miRNAs, and in 

particular we have identified mir-150-5p as a regulator of the 

myofibroblast phenotype and potential biomarker of fibrosis in 

ALD patients. 

Disclosures: 

The following authors have nothing to disclose: Luigi Locatelli, Pier P. Paoli, Timothy 

Hardy, Rachel M. Howarth, Fiona Oakley, Derek Mann 

1409 

Human precision Cut Liver Slices model for testing 

anti-fibrotic drugs 

Lynda Aoudjehane 1,2 , Margaux Legrand 1 , Grégoire Bisch 1 , Rolland 

Delelo 2 , Chantal Housset 2 , Pierre Balladur 3 , Claire Goumard 

4 , Jérôme Becquart 1 , Yvon Calmus 2,5 , Filomena Conti 2,5 ; 

1 Human HepCell, Faculté de Médecine Pierre et Marie Curie, 

Paris, France; 2 UMPC UNIV Paris 6 & INSERM, UMR_S 938, 

CDR Saint-Antoine, Paris, France; 3 AP-HP, Hôpital Saint Antoine, 

Service de Chirurgie Digestive, Paris, France; 4 AP-HP, Hôpital Pitié- 

Salpêtrière, Service de Chirurgie Digestive, Paris, France; 5 AP- 

HP, Hôpital Pitié-Salpêtrière, Unité de Transplantation Hépatique, 

Paris, France 

Background: Fibrosis is the common end stage of all chronic 

liver diseases. Understanding the molecular mechanisms underlying 

liver fibrogenesis is mandatory to develop new antifibrotic 

therapies. Current methods for evaluating the efficacy 

of anti-fibrotic drugs rely mostly on immortalized cell lines and 

animal models. Human precision-cut liver slices (PCLS) preserve 

the cell types and proportions, and particularly the cell-cell 

interactions. Several studies have recently suggested that PCLS 

are potentially useful as in vitro model to study liver fibrosis. 

The current study aimed to test the efficacy of human PCLS to 

evaluate the effect of anti-fibrotic drugs. Methods: Human PCLS 

of 250 μm thickness were obtained from normal or cirrhotic 

liver samples. PCLS were incubated for 1-48h at 37°C under 

85% O2 and, after 24h, were treated with different anti-fibrotic 

drugs: ciclosporin A (CsA), pirfenidone (PFD) or SMAD3 

inhibitor (SIS3). The viability of PCLS was assessed by ATP production. 

Hepatic metabolism and fibrosis markers (α-SMA and 

Coll1, HSP47, TIMP-1, MMP-2 and TGF-beta) were determined 

by RT-PCR. Results: PCLS from normal and cirrhotic human liver 

samples could be cultured with an excellent viability over 48h 

and this culture induced a spontaneous fibrosis. The expression 

of fibrotic markers (α-SMA and Coll1, HSP47, TIMP-1, 

MMP-2 and TGF-beta) significantly increased at 24 and 48h 

when compared to 1h. Fibrosis markers significant decreased 

(α-SMA and Coll1 and TIMP1 mRNA expression) when normal 

or cirrhotic human PCLS were treated for 24h with PFD 

and SIS3. However, CsA had no significant effect on fibrosis 

marker expression on normal or cirrhotic PCLS. Conclusion: 

These preliminary results suggest that human PCLS can be 

maintained in culture for 48 hours without significant modification 

of viability. The culture of PCLS induces a spontaneous 

activation of the fibrotic process. This feature can be used to 

study the effect of antifibrotic molecules and thus constitutes 

an interesting physiological human model for the screening of 

antifibrotic drugs. 

Disclosures: 

Jérôme Becquart - Management Position: Human Hepcell 

The following authors have nothing to disclose: Lynda Aoudjehane, Margaux Legrand, 

Grégoire Bisch, Rolland Delelo, Chantal Housset, Pierre Balladur, Claire 

Goumard, Yvon Calmus, Filomena Conti 

1410 

The AMPK-related kinase NUAK2 interacts with TGF-β 

and regulates the activation of hepatic stellate cells 

Cristina Tosti Guerra 1 , Alessandra Caligiuri 1 , Angela Provenzano 1 , 

Krista Rombouts 2 , Massimo Pinzani 2 , Fabio Marra 1 ; 1 University 

of Florence, Florence, Italy; 2 University College London, London, 


Background and Aims: Nuak2 is a member of AMPK related 

kinases (ARKs), that act as energy sensors and controllers of 

cellular structure, with different effects on cell motility and cytoskeletal 

organization, depending on the cell types. AMPK pos-


sesses anti-fibrogenic activity. A recent study showed a link 

between Nuak2 and TGF-β, a major driver of hepatic fibrogenesis. 

However, little information is available on the role 

of Nuak2 in liver fibrosis, and in particular on HSC trans-differentiation. 

Aims of this study was to elucidate the involvement 

of Nuak2 in HSC transactivation, and to investigate the 

possible interaction between Nuak2 and TGF-β signaling. 

Methods: HSC were isolated from normal rat and human livers 

and activated by culture on plastic. Knockdown of Nuak2 was 

achieved by siRNA. Cell migration was evaluated in modified 

Boyden Chambers. Protein expression and signaling pathways 

were analyzed by Western blotting. Results: In fully activated 

HSC, down-regulation of Nuak2 positively modulated cell 

migration and induced changes in the expression of molecules 

involved in cytoskeletal organization. Knockdown of Nuak2 

increased α-SMA expression and phosphorylation of SMAD3, 

in HSC exposed to TGFβ. Moreover, Nuak2 expression was 

up-regulated in HSC following TGFβ treatment. Conclusions: 

The AMPK related kinase, Nuak2, is modulated during the activation 

process of HSC, regulates cytoskeletal organization and 

cell motility and interacts with TGF-β in regulating the pro-fibrogenic 

properties of HSC. 

Disclosures: 



Gilead, BMS 

Fabio Marra - Advisory Committees or Review Panels: Abbvie; Consulting: Bayer 

Healthcare; Grant/Research Support: ViiV 

The following authors have nothing to disclose: Cristina Tosti Guerra, Alessandra 

Caligiuri, Angela Provenzano, Krista Rombouts 

1411 

The bile acid-phospholipid conjugate Ursodeoxycholyl 

Lysophosphatidylethanolamide (UDCA-LPE) disturbs 

pro-fibrogenic Integrin and TGFβ signaling 

Jie Su, Hongying Gan-Schreier, Walee Chamulitrat, Wolfgang 

Stremmel, Anita Pathil; Department of Internal Medicine IV, University 

of Heidelberg, Heidelberg, Germany 

BACKGROUND: Integrin receptors, which are involved in cellcell 

and cell-matrix interaction emerge as crucial mediators of 

TGFβ1 activation in liver fibrosis. Ursodeoxycholyl Lysophosphatidylethanolamide 

(UDCA-LPE) is a synthetic bile acid-phospholipid 

conjugate with hepatoprotective and anti-fibrogenic 

functions in vitro and in vivo. In this study we aim to elucidate 

signaling pathways, which mediate anti-fibrogenic action of 

UDCA-LPE. RESULTS: In order to promote pro-fibrogenic signaling 

upon extracellular matrix binding integrins recruit focal 

adhesion kinase (FAK) and SRC kinase, which are phosphorylated 

in response to integrin engagement. Incubation of CL48 

liver cells with UDCA-LPE altered the localization of integrin α2, 

α3, α5, αv, β1, β4, β5 and β6 as observed by immunofluorescence. 

In contrast, incubation with the integrin blocking peptide 

RGD reduced integrin protein levels, but did not change its 

localization. After UDCA-LPE treatment integrins were transported 

to the ER and the nuclear envelop, which led to a loss of 

co-localization of integrins with SRC resulting in dephosphorylation 

of FAK (Tyr 925 and Tyr 576/577) and SRC (Tyr416). 

This translocalization of integrins was not induced by UDCA 

and/or LPE treatment, but was exclusively achieved by UDCA- 

LPE. To further dissect whether UDCA-LPE would mediate a 

shift of integrins to certain membrane microdomains such as 

lipid raft and non-raft regions lipid fractioning was performed. 

Integrin α2, α3, α5, αv, β1, β4 and β6, but not SRC, was 

significantly reduced in transitional fractions and increased in 

lipid raft fractions. Taken together, UDCA-LPE disturbed integrin-FAK 

signaling after short activation by modifying the localization 

of integrins. UDCA-LPE further led to a shift of TGFβ1 

receptor I and II to the ER and the nuclear envelope resulting 

in dephosphorylation of Smad2/3. According to lipid fractionation 

TGFβ1 receptor I and II were also reduced in transitional 

fractions and increased in lipid raft fractions after UDCA-LPE 

treatment. Analysis by HPLC-MS revealed that UDCA-LPE shows 

an integrated fractionation of UDCA and LPE, suggesting that 

the localization of UDCA-LPE depends on both UDCA and LPE 

end. CONCLUSIONS: UDCA-LPE mediated translocation of 

integrins and TGFβ1 receptors into lipid rafts leads to a loss of 

colocalization with their down-stream signaling proteins SRC 

and Smad2/3. By inhibiting crucial pro-fibrogenic signaling 

pathways UDCA-LPE emerges as a promising experimental 

drug-candidate for the treatment of liver fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Jie Su, Hongying Gan-Schreier, 

Walee Chamulitrat, Wolfgang Stremmel, Anita Pathil 

1412 

Curcumin blocks the loss of lipid droplets in activated 

hepatic stellate cells by increasing perilipin 5 gene 

expression in vitro 

DingYu Zhang; Hepatology, Wuhan Medical Treatment Center, 

Wuhan, China 

BACKGROUND: Activation of hepatic stellate cells(HSCs) are 

the major players during liver fibrogenesis, and featured by the 

loss of intracellular lipid droplets(LD). Currently, accumulating 

evidence showed that blockade of LD loss could inhibit HSC 

activation including decreasing cell proliferation and extracellular 

matrix production. Curcumin, a phytochemical from 

turmeric, has been demonstrated to inhibit HSC activation and 

protect the liver from fibrogenesis in vitro and in vivo. However, 

whether curcumin affects intracellular LD formation in 

HSCs and underlying mechanism remain largely undefined. 

AIM AND HYPOTHESIS: The aims of this study are to evaluate 

roles of curcumin in the formation of LD in HSC, and to further 

explore the underlying mechanisms. It is hypothesized that curcumin 

might eliminate the loss of LD in activated HSCs, rather 

than hepatocytes, by increasing perilipin 5(plin5) gene expression 

and inducing lipogenesis. RESULTS: Curcumin restored 

intracellular LD formation, and increased triglyceride (TG) and 

free fatty acid(FFA) levels as well as induced expression of 

genes closely relevant to lipogenesis including SREBP-1c, fatty 

acid synthase(FAS), PPAR-coactivator (PCA), in passaged mice 

HSCs. In contrast, under the same condition, the LD formation 

and lipid contents have been unaffected by curcumin in 

mice hepatocytes. Furthermore, curcumin dose-dependently 

increased plin5 gene expression which has been suppressed 

in activated state in HSCs. Meanwhile, transduction with lentiplin5-YFP 

in passaged HSCs had the similar effect as curcumin 

treatment. On the contrary, in HSCs transfected with plin5- 

siRNA prior to curcumin treatment, LD contents and lipid levels 

were not changed significantly compared with the normal cells. 

CONCLUSIONS: Our results demonstrate that curcumin restores 

the LD formation in activated HSCs in vitro, which is carried 

out by inducing lipid synthesis via increasing lipogenesis-related 

gene expression. The process is mediated by activating 

plin5 gene expression. Our results provide novel insight into 

the mechanism of the role of curcumin in the inhibition of HSC 

activation and hepatic fibrogenesis and potential therapeutic 

strategies for treatment of hepatic fibrogenesis without increasing 

lipotoxicity in hepatocytes. 

Disclosures: 

The following authors have nothing to disclose: DingYu Zhang


1413 

Versican: A Novel Modulator of Hepatic Fibrosis 

Terence N. Bukong 1 , Sean B. Maurice 1,2 , Barinder Chahal 2,1 , 

David Schaeffer 2 , Paul J. Winwood 2,1 ; 1 Northern Medical Program, 

University of Northern BC, Prince George, BC, Canada; 

2 University of British Columbia, Vancouver, BC, Canada 

Little is known about the deposition and turnover of proteoglycans 

in liver fibrosis, despite their abundance in the extracellular 

matrix. Versican plays diverse roles in modulating cell 

behaviour in other fibroproliferative diseases, but remains 

poorly described in the liver. Hepatic fibrosis was induced by 

carbon tetrachloride (CCl 4 

) treatment of C57BL/6 mice over 

4 weeks followed by recovery over a 28 day period. Primary 

mouse hepatic stellate cells (HSCs) were activated in culture 

and versican was transiently knocked down in human (LX2) 

and mouse HSCs. Expression of versican, A Disintegrin-like and 

Metalloproteinase with Thrombospondin-1 motifs (ADAMTS)-1, 

-4, -5, -8, -9, -15 and -20, and markers of fibrogenesis were 

studied using qRT-PCR and Western blotting. CCl 4 

treatment 

led to significant increases in versican expression and the proteoglycanases 

ADAMTS-5, -9, -15 and -20, alongside TNF-α, 

alpha-smooth muscle actin (α-SMA), collagen-1, and TGF-β 

expression. During recovery, expression of many of these genes 

returned to control levels. However, expression of ADAMTS-5, 

-8, -9 and -15 showed delayed increases in expression at 28 

days of recovery which corresponded with decreases in versican 

V0 and V1 cleavage products (G1-DPEAAE 1401 and 

G1-DPEAAE 441 ). Activation of primary HSCs in-vitro significantly 

increased versican, α-SMA, and collagen-1 expression. 

Transient knockdown of versican in HSCs led to decreases in 

markers of fibrogenesis and reduced cell proliferation, without 

inducing apoptosis. Versican expression increases during 

HSC activation and liver fibrosis, and proteolytic processing 

occurs during the resolution of fibrosis. Knockdown studies in 

vitro suggest a possible role of versican in modulating hepatic 

fibrogenesis. 

Disclosures: 

The following authors have nothing to disclose: Terence N. Bukong, Sean B. 

Maurice, Barinder Chahal, David Schaeffer, Paul J. Winwood 

1414 

Hepatic stellate cell-derived retinoid enhances functional 

maturity of human embryonic stem cell-derived hepatocytes 

on drug metabolism 

Hyuk-Soo Eun, Wonhyo Seo, Won-IL Jeong; KAIST, Daejeon, 


Background: Human embryonic stem cell-derived hepatocyte 

(hES-Hep) could be used for the screening tool of drug hepatotoxicity. 

However, drug metabolic functions of hES-Hep 

are still poor. Retinoids including retinol and its metabolites 

play important roles not only in organ development but also 

metabolic regulation and they are enriched in hepatic stellate 

cells (HSCs) of liver. Therefore, in this study, we evaluate the 

effects of mouse HSCs (mHSCs) on the expression and activity 

of drug metabolic enzymes in hES-Hep. Methods: Analyses 

of microarray, co-culturing with mHSCs or human HSC cell 

lines (LX-2 and hTERT), retinoid treatment, flow cytometry analysis, 

protein and gene expression, and drug metabolic activity 

were performed in hES-Hep. Results: In microarray analyses, 

gene expression of drug metabolic enzymes such as CYP1A2, 

CYP2C9, CYP2D6, CYP2E1 and CYP3A4 in hES-Hep was 

specifically down-regulated compared with normal hepatocyte. 

However, that expression was significantly increased in 

co-cultured hES-Hep with retinol-storing mHSCs compared to 

control hES-Hep. In co-cultured mHSCs, expression of LRAT was 

down-regulated while expression of retinaldehyde dehydrogenase-1 

(Raldh1) and cellular retinoic acid binding protein-1 

was significantly increased, indicating decreased retinol storage 

but increased production and delivery of retinoic acids in 

HSCs. Similarly, direct treatments of retinoic acids increased 

expression of metabolic enzymes in hES-Hep compared with 

control hES-Hep. In contrast, retinoic acid deficiency in LX-2, 

hTERT and Raldh1-depleted HSCs did not affect gene expression 

of metabolic enzymes in hES-Hep, suggesting that retinoic 

acid might be a critical factor for functional maturation of hES- 

Hep. In flow cytometry, two biggest populations of hES-Hep 

depending on cell size and granularity expressed major types 

of drug metabolic enzymes and one smallest population still 

highly expressed mesodermal markers such as SOX17, FOXA2 

and GATA4. In western blotting, protein levels of CYP1A2, 

CYP2E1, and CYP3A4 were increased in co-cultured hES-Hep 

with retinol-storing HSCs. Consistently, metabolic activities of 

CYP1A2 (phenacetin), CYP3A4 (testosterone) and CYP2E1 

(chlorozoxazone) were significantly improved in co-cultured or 

retinoic acid-treated hES-Hep compared with controls. Conclusions: 

Based on our data, retinol-storing mHSCs could enhance 

functional maturation of hES-Hep through retinoic acid-mediated 

increased expression and activity of drug metabolic 

enzymes. Thus, co-culturing with mHSCs or treatments of retinoic 

acids could be useful ways to adapt immature hES-Hep for 

the application of drug toxicity screening system. 

Disclosures: 

The following authors have nothing to disclose: Hyuk-Soo Eun, Wonhyo Seo, 

Won-IL Jeong 

1415 

Calcium Mobilization through L-type Channels in 

Hepatic Stellate Cell is Essential for TGF-b-mediated 

CTGF Induction in Pyk2-dependent Manner 

Jonghwa Kim 1 , SoHee Kang 1 , Soohyun Park 1 , Ju-Yeon Cho 1 , Won 

Sohn 1 , David A. Brenner 2 , Yong-Han Paik 1 ; 1 Samsung medical 

center, Seoul, Korea (the Republic of); 2 UC San Diego, La Jolla, 

CA 

Background: In hepatic fibrogenesis hepatic stellate cell (HSC) 

is a major cell type responsible for producing a major profibrogenic 

cytokine TGF-b, and connective tissue growth factor 

(CTGF), a major fibrogenic mediator in several organs. The 

multi-functional nature of TGF-b signaling in hepatic fibrogenesis 

is still elusive. At the previous AASLD (2014) we reported 

that Pyk2 is essential for TGF-b-mediated, Smad-independent 

CTGF induction. Pyk2 is known to be calcium-sensitive, and 

TGF-b was reported to increase intracellular calcium level. 

Therefore, we investigated if TGF-b activates Pyk2 by increasing 

intracellular calcium levels through L-type voltage-gated 

calcium channel in hepatic stellate cell. Methods: Immortalized 

human stellate cell line, LX-2, has been cultured. After TGF-b 

treatment, expression of CTGF and a-SMA were assessed with 

RT-PCR and western blot. Pharmacological inhibitor and siR- 

NA-mediated knockdown were used to modulate the activities 

and expression levels of protein. Intracelllular calcium mobilization 

was measured with Fura-2/AM. Activation of Pyk2 

was addressed in western blot using different phosphorylation 

site-specific antibodies. Results: CTGF expression was up-regulated 

within 1hr in TGF-b stimulated LX-2. This up-regulation 

was greatly suppressed by siRNA-mediated knockdown and 

pharmacological inhibitor of Pyk2. TGF-b treatment increased 

phosphorylation of Pyk2 on tyrosine 402, 579/580, and 881. 

Consistent with the previous reports, TGF-b increased intracellular 

calcium concentration in Fura-2-preloaded LX-2. CTGF


induction by TGF-b was blocked in dose-dependent manner 

by pre-treatment with BAPTA-AM, an intracellular calcium chelator. 

Treatment of LX-2 with A23187, a calcium ionopore, 

increased CTGF induction in the absence of TGF-b, suggesting 

that increase of intracellular calcium level is enough to induce 

CTGF expression. In addition, A23187 increased activation of 

Pyk2 in western. The CTGF induction by A23187 is also greatly 

reduced by siRNA of Pyk2. Pre-treament of LX-2 with Nifedipine 

(an L-type calcium channel blocker) suppressed CTGF induction 

by TGF-b in dose-dependent manner, while FPL64176 (L-type 

channel activator) increased CTGF expression without TGF-b. 

The CTGF up-regulations by TGF-b, A23187, and FPL64176 

were all suppressed by siRNA-mediated knockdown and pharmacological 

inhibitors of Pyk2. Conclusions: In hepatic stellate 

cell, TGF-b increases the intracellular calcium level through 

L-type calcium channel, leading to downstream Pyk2 signaling 

for CTGF induction. 

Disclosures: 

The following authors have nothing to disclose: Jonghwa Kim, SoHee Kang, 

Soohyun Park, Ju-Yeon Cho, Won Sohn, David A. Brenner, Yong-Han Paik 

1416 

Paracrine suppression of proliferation and activation of 

hepatic myofibroblasts by adipose-derived stem cells 

Danial Afsharzadeh 2 , Shiva Shajari 1 , Linda Brouwer 2 , Han 

Moshage 1 , Martin C. Harmsen 2 , Klaas Nico Faber 1 ; 1 MDL, University 

medical center Groningen, Groningen, Netherlands; 2 Medical 

Biology, University Medical Center Groningen, Groningen, 

Netherlands 

BACKGROUND AND AIM: Chronic liver diseases lead to liver 

fibrosis that may progress to cirrhosis and predisposes for liver 

cancer. Liver fibrosis is the result of excessive extracellular 

matrix production by hepatic myofibroblasts that may originate 

from hepatic stellate cells (HSC) and portal myofibroblasts 

(PMF). Though fibrosis is reversible, no drug-based therapy is 

available to cure fibrosis and liver transplantation is the only 

life-saving option for patients with advanced cirrhosis. Transplantation 

of mesenchymal stem cells has been shown to significantly 

improve liver function in end-stage liver cirrhosis. Among 

mesenchymal stem cells, adipose-derived stem cells (ADSC) 

are especially attractive in the context of future clinical applications. 

Indeed, recent studies demonstrate that ADSC ameliorate 

chemical- and diet-induced liver fibrosis, but the mechanistic 

understanding is limited. It remains to be determined whether 

ADSC produced factors that act in a paracrine fashion or that 

the ADSCs engraft and differentiate to functional liver cells. 

Here, we investigated the interaction between human ADSC 

and primary rat HSC and PMF. METHODS: Human ADSC were 

isolated from Human subcutaneous adipose tissue and primary 

rat HSC and PMF from Wistar rats. ADSC and HSC or PMF 

were cocultured in a transwell system. Moreover, HSC and 

PMF were cultured in conditioned medium from ADSC (24 

h culture). Cell proliferation and migration was assessed by 

real time cell analysis (xCELLigence). Expression of markers 

of fibrosis, e.g. Col1a1 and Acta2, was analyzed by Q-PCR. 

Chemokine receptor antagonists (SB225002, NBI74330 and 

AMD 3465) were used to block CXCR2, CXCR3 and CXR4, 

respectively. RESULTS: ADSC showed a strong migratory 

potential towards activated HSC and PMF, but not to quiescent 

HSC. None of the individual chemokine receptor antagonists 

affected ADSC migration towards HSC/PMF. Only the combination 

of all 3 antagonists showed a partial inhibition of the 

migratory phenotype of ADSC. Proliferation of activated HSC 

and PMF, as well as the expression of Col1a1 and Acta2, was 

suppressed when cocultured with ADSC. A similar effect was 

observed when monocultures of activated HSC and PMF were 

exposed to conditioned medium of ADSC. CONCLUSIONS: 

ADSC are highly migratory towards hepatic myofibroblasts 

through a combination of chemokine signaling pathways. 

ADSC produce factors that suppress proliferation and activation 

of hepatic myofibroblasts in a paracrine manner. Identifying 

the antifibrotic factor(s) in the ADSC secretome holds 

promise for novel antifibrotic therapies. 

Disclosures: 

The following authors have nothing to disclose: Danial Afsharzadeh, Shiva Shajari, 

Linda Brouwer, Han Moshage, Martin C. Harmsen, Klaas Nico Faber 

1417 

WITHDRAWN 

1418 

Deactivation of human and rat cirrhotic hepatic stellate 

cells: a novel alternative use for the glucagon-like 1 

receptor agonist Liraglutide. 

Fernanda C. de Mesquita 1,2 , Sergi Guixé-Muntet 1 , Jose Luis 

Rosa 3 , Jaime Bosch 1 , Jarbas R. de Oliveira 2 , Jordi Gracia-Sancho 

1 ; 1 Barcelona Hepatic Hemodynamic Lab, IDIBAPS - Hospital 

Clinic de Barcelona - CIBEREHD, Barcelona, Spain; 2 Laboratório 

de Biofísica Celular e Inflamação, PUCRS, Porto Alegre-RS, Brazil; 

3 Departament de Ciències Fisiològiques II, Institut d’Investigació 

Biomèdica de Bellvitge (IDIBELL), University of Barcelona, L’Hospitalet 

de Llobregat, Spain 

Background and Aims: Activation of hepatic stellate cells 

(HSC) plays a key role in hepatic fibrogenesis, thus improving 

HSC phenotype may help to promote fibrosis regression 

and ameliorate chronic liver disease. Glucagon-like peptide 1 

(GLP-1) receptor agonists, like liraglutide, are well established 

in the control of type 2 diabetes. Considering recent studies 

demonstrating anti-inflammatory and anti-oxidant properties 

to this kind of drugs, our study aimed to evaluate the effects of 

liraglutide on activated HSC phenotype. Methods: Liraglutide 

(10-50mM), or its vehicle, was administered to human and rat 

cirrhotic HSC (24h and 72h). HSC activation phenotype was 

assessed by mRNA and protein expression of alpha smooth 

muscle actin (α-SMA) and type I collagen. Effects of liraglutide 

on HSC viability and proliferation were evaluated by cell counting 

and platelet-derived growth factor receptor (PDGFR) expression. 

Results: Liraglutide markedly improved HSC phenotype as 

demonstrated by the down-regulation of a-sma (-40% mRNA, 

-40% protein expression vs. vehicle) and collagen (-60% mRNA 

expression) after 72 hours of treatment. Liraglutide did not 

affect HSC viability but significantly diminished cell proliferation 

(-32% in cell proliferation, -50% in PDGFR expression vs. 

vehicle-treated cells). All p


1419 

Serine Protease Omi/HtrA2 Reduces Oxidative Stress 

and Preserves Mitochondrial Function in Liver Fibrosis 

Seung Kew Yoon 1,2 , Wonhee Hur 1 , Joon Ho Lee 1 , Sung Woo 

Hong 1 , Jung-Hee Kim 1 , Jung Eun Choi 1 , Sung Min Kim 1 , Eun Byul 

Lee 1 ; 1 The Catholic University Liver Research Center & WHO Collaborating 

Center of Viral Hepatitis, Seoul, Korea (the Republic 

of); 2 Department of Internal Medicine, St. Mary’s Hospital, The 

Catholic University of Korea, Seoul, Korea (the Republic of) 

Background: Liver fibrosis is one of chronic liver diseases 

caused by various causes. A mitochondrial serine protease 

Omi/high temperature requirement protein A2 (HtrA2) is 

involved in several disease, but has been poorly understood 

in relation to liver fibrosis. The aim of this study is to investigate 

the role of Omi/HtrA2 in the liver fibrosis. Methods: Protein 

expression of Omi/HtrA2 was analyzed with liver tissues 

from CCl - 4 

induced mice and liver cirrhosis patients. Reactive 

oxygen species (ROS) damage was measured by H 2 

DCF-DA 

and mito-Sox staining in isolated hepatocytes from mnd2 mice 

which have the S276C mutation in the protease domain of 

Omi/HtrA2. The Omi/HtrA2 gene was delivered by hydrodynamic 

injection in the CCl 4 

induced-liver fibrosis mice model 

which had lower Omi/HtrA2 expression. The change of collagen 

accumulation was measured by Masson’s trichrome (MT) 

staining and hydorxyproline assay. Morphological alteration 

of mitochondria was analyzed by using transmission electron 

microscopy (TEM). Result: Protein expression of Omi/HtrA2 

was decreased in liver tissues from CCl 4 

induced mice and 

liver cirrhosis patients. In mnd2 mice, oxidative stress increased 

compared with wild type mice and abnormal morphology of 

mitochondria was observed under TEM. Mitochondrial damage 

induced with CCl 4 

was protected by hydrodynamic based 

gene delivery of Omi/HtrA2. In addition, the overexpression 

of Omi/HtrA2 decreased collagen accumulation in the liver 

tissue. Conclusion: Our findings suggest that Omi/HtrA2 

overexpression modulates mitochondrial ROS generation in 

liver fibrosis. Omi/HtrA2 gene delivery in liver fibrosis mice 

model showed anti-oxidative effect through preserving liver 

mitochondrial functions. Therefore, the modulation of Omi/ 

HtrA2 through the mitochondrial antioxidant defense mechanism 

might be promising in anti-fibrotic therapeutic approach. 

Disclosures: 

The following authors have nothing to disclose: Seung Kew Yoon, Wonhee Hur, 

Joon Ho Lee, Sung Woo Hong, Jung-Hee Kim, Jung Eun Choi, Sung Min Kim, 

Eun Byul Lee 

1420 

Cancer associated fibroblasts in intrahepatic cholangiocarcinoma 

is derived from portal fibroblasts, but 

not hepatic stellate cells; immunohistochemical study in 

human tissues 

Naoki Uyama 1 , Rei A. Ito 1 , Ami Kurimoto 1 , Tomohiro Okamoto 1 , 

Akito Yada 1 , Hideaki Sueoka 1 , Seikan Hai 1 , Hisashi Kosaka 1 , 

Yuichi Kondo 1 , Ikuo Nakamura 1 , Kazuhiro Suzumura 1 , Yasukane 

Asano 1 , Toshihiro Okada 1 , Tadamichi Hirano 1 , Junichi 

Yamanaka 1 , Norifumi Kawada 2 , Jiro Fujimoto 1 ; 1 Hepato-biliary-pancreas 

surgery, Hyogo College of Medicine, Nishinomiya, 

Japan; 2 Department of Hepatology, Osaka City University Graduate 

School of Medicine,, Osaka, Japan 

(Background and aim) Cancer associated fibroblasts (CAFs), 

a key player in the extracellular matrix production of cancer 

tissue, is known to be involved in the regulation of cancer 

behavior. Although origin of CAFs in intrahepatic cholangiocarcinoma 

(ICC) is suspected to be composed of hepatic 

stellate cells (HSCs), portal fibroblasts (PF), or bone marrow 

derived cells such as fibrocytes, detailed analysis has lacked 

so far. Until now, it has been reported that fascin is a specific 

marker for HSC and fibrin-2 and Thy-1 are specific markers 

for PFs, while α-SMA and PDGFRβ are expressed by both 

cells. However, there has been no report on the expression 

of PDGFRα in hepatic mesenchymal cells. In this study, we 

performed immunohistochemistry in order to characterize 

CAFs in human ICC. (Materials and Methods) Ten pairs of ICC 

cancerous and noncancerous tissues were prepared. Tissue 

sections were Zinc-fixed and paraffin-embedded. Immunohistochemistry 

of α-SMA, PDGFRα, PDGFRβ, fascin, fibrin-2 and 

Thy-1 in cancerous and noncancerous area was performed. 

As for noncancerous area, peri-portal area and sinusoidal 

area were intensively observed. To investigate the expression 

of individual markers in PFs, HSCs and CAF, double-labeled 

immunofluorescent staining with α-SMA was performed under 

confocal microscope. (RESULTS) In immunohistochemistry of 

noncancerous area of ICC, PDGFRα expression was detected 

at periportal area, but not in sinusoidal area. Double immunofluorescent 

staining for PDGFRα and α-SMA revealed that both 

protein expressions were overlapped in PFs, but not in HSCs, 

indicating PDGFRα as a novel marker for PFs. Further immunohistochemical 

analyses showed PFs are characterized as 

α-SMA + , PDGFRβ + , fascin - , fibrin-2 + and Thy-1 + and HSCs are 

as α-SMA + , PDGFRβ + , fascin + , fibrin-2 - and Thy-1 - . In cancerous 

area, immunohistochemistry revealed that cancer stroma 

was positive for all of α-SMA, PDGFRα, PDGFRβ, fibrin- and 

Thy-1. As for fascin, 3 samples were positive, but the others 

were negative. α-SMA expression was overlapped with any of 

PDGFRα, PDGFRβ, fibrin-2 or Thy-1 in CAFs. (Conclusion) Our 

human study revealed that PDGFRα is a specific marker for 

PFs. HSCs are α-SMA + , PDGFRβ + , fascin + , fibrin-2 - , Thy-1 - and 

PDGFRα - , while PFs are α-SMA + , PDGFRβ + , fascin - , fibrin-2 + , 

Thy-1 + and PDGFRα + . CAFs associated with ICC are positive 

for PF markers (fibrin-2 + , Thy-1 + and PDGFRα + ) in addition to 

α-SMA and PDGFRβ. Taken together, it is plausible that CAFs 

are derived from PFs rather than HSCs. 

Disclosures: 



The following authors have nothing to disclose: Naoki Uyama, Rei A. Ito, Ami 

Kurimoto, Tomohiro Okamoto, Akito Yada, Hideaki Sueoka, Seikan Hai, Hisashi 

Kosaka, Yuichi Kondo, Ikuo Nakamura, Kazuhiro Suzumura, Yasukane Asano, 

Toshihiro Okada, Tadamichi Hirano, Junichi Yamanaka, Jiro Fujimoto 

1421 

Human neutrophil peptide-1 enhances alcohol-induced 

hepatocyte apoptosis in vivo and in vitro. 

Rie Ibusuki 2,1 , Hirofumi Uto 2,3 , Masaya Kozono 2 , Kohei Oda 2 , 

Akihiko Oshige 2 , Kotaro Kumagai 2 , Seiichi Mawatari 2 , Tsutomu 

Tamai 2 , Akihiro Moriuchi 2 , Hirohito Tsubouchi 4,5 , Takezaki 

Toshiro 1 , Akio Ido 2,4 ; 1 Department of International Island and 

Community Medicine, Kagoshima University Graduate School of 

Medical and Dental Sciences, Kagoshima, Japan; 2 Digestive and 

Lifestyle Diseases, Department of Human and Environmental Science, 

Kagoshima University Graduate School of Medical and Dental 

Sciences, Kagoshima, Japan; 3 Center for Digestive and Liver 

Diseases, Miyazaki Medical Center Hospital,, Miyazaki, Japan; 

4 Department of HGF Tissue Repair and Regenerative Medicine, 

Kagoshima University Graduate School of Medical and Dental Sciences, 

Kagoshima, Kagoshima, Japan; 5 Kagoshima City Hospital,, 

Kagoshima, Japan 

Background Neutrophil infiltration of the liver is a typical 

feature of alcoholic liver injury and nonalcoholic steatohepatitis. 

Human neutrophil peptide (HNP)-1 is an antimicrobial 

peptide secreted by neutrophils. We previously reported that


transgenic expression of HNP-1 enhances hepatic fibrosis 

and hepatocyte apoptosis in a murine steatohepatitis model 

established by ethanol administration or a choline-deficient, 

L-amino acid–defined diet. We also showed that HNP-1 exerts 

an additive effect on ethanol-induced hepatocyte apoptosis in 

SK-Hep1 hepatocellular carcinoma cells in vitro. The aim of 

this study was to elucidate the mechanism of hepatocyte apoptosis 

induced by HNP-1. Materials and Methods Transgenic 

(Tg) mice expressing HNP-1 under the control of a β-actin– 

based promoter were established. Ethanol was orally administered 

to HNP-1 Tg or wild-type C57BL/6N (WT) mice for 24 

weeks. Gene expression and protein levels in liver tissue were 

evaluated by RT-PCR and Western blot analysis, respectively. 

Apoptosis-related microRNAs were analyzed using miScript 

miRNA PCR Array and real-time PCR. SK-Hep1 hepatocellular 

carcinoma cells were used to investigate the effect of HNP-1 

on hepatocytes in vitro. Results After 24 weeks of ethanol 

administration, hepatocyte apoptosis, as assessed by TUNEL 

assay, was significantly more severe in TG mice than in WT 

mice. Apoptosis was accompanied by higher protein levels of 

caspase-3, caspase-8, cleaved PARP, and Bax in liver tissue. 

Levels of CD14, TLR4, NFκB-p65, and IL-6 were higher in TG 

mice than in WT mice. In addition, p-ASK1, ASK1, p-JNK, 

JNK1, JNK2, and CHOP were all more abundant in TG mice 

than in WT mice. By contrast, the level of anti-apoptotic Bcl-2 

protein in the liver was significantly lower in TG mice than 

in WT mice. Analysis of apoptosis-related microRNAs in liver 

tissue revealed that miR-34a-5p expression was significantly 

higher in TG mice than in WT mice. Furthermore, in the presence 

of ethanol, HNP-1 increased the levels of caspase 3 and 

Bax and decreased the level of Bcl-2 in a concentration-dependent 

manner in vitro. Conclusion HNP-1 secreted by neutrophils 

may exacerbate hepatocyte apoptosis following alcoholic liver 

injury via elevated microRNA expression. 

Disclosures: 

The following authors have nothing to disclose: Rie Ibusuki, Hirofumi Uto, Masaya 

Kozono, Kohei Oda, Akihiko Oshige, Kotaro Kumagai, Seiichi Mawatari, 

Tsutomu Tamai, Akihiro Moriuchi, Hirohito Tsubouchi, Takezaki Toshiro, Akio Ido 

1422 

Liver Fibrosis in a Mouse Model of Non-alcoholic Fatty 

Liver Disease is Mediated by Hepatocyte Hypoxia Inducible 

Factor-1 

Omar Mesarwi, Mi-Kyung Shin, Shannon Bevans-Fonti, Vsevolod 

Polotsky; Johns Hopkins University, Baltimore, MD 

Introduction: Obstructive sleep apnea (OSA) is associated with 

the progression of non-alcoholic fatty liver disease (NAFLD) to 

steatohepatitis and fibrosis. This progression correlates with the 

severity of OSA and, specifically, with the burden of hypoxia. 

In mice with diet induced obesity, hepatic steatosis leads to 

liver tissue hypoxia, which is worse with exposure to intermittent 

hypoxia. Intermittent hypoxia in mice also results in more 

severe liver fibrosis. We hypothesized that hepatocyte specific 

knockout of the oxygen sensing α subunit of hypoxia inducible 

factor-1 (HIF-1), a master regulator of the global response to 

hypoxia, may be protective against the development of liver 

fibrosis. Methods: Hepatocytes were isolated from wild-type 

C57BL6/J mice and were exposed to sustained hypoxia (1% 

O 2 

) or normoxia (16% O 2 

) for 24 hours. The culture media 

was used to reconstitute type I collagen and the resulting 

matrices were examined using confocal microscopy. This was 

repeated using hepatocytes from mice with hepatocyte-specific 

HIF-1α knockout (Hif1a -/- hep mice). Additionally, wild-type 

(n=7) and Hif1a -/- hep (n=10) mice were fed a high trans-fat 

diet for six months, to induce hepatic steatosis. Hepatic fibrosis 

was evaluated by Sirius red stain and collagen quantification 

by hydroxyproline assay. Liver enzymes, fasting insulin, 

and hepatic triglyceride content were also assessed. Results: 

Culture media from wild-type mouse hepatocytes exposed to 

hypoxia allowed for avid collagen cross-linking, but very little 

cross-linking was seen when hepatocytes were exposed to normoxia, 

or when hepatocytes from Hif1a -/- hep mice were used 

in hypoxia or normoxia. Wild-type mice on a high trans-fat diet 

had 80% more hepatic collagen than Hif1a -/- hep mice (2.21 

mg collagen/mg liver tissue, versus 1.23 mg collagen/mg liver 

tissue, p=0.03), which was confirmed by Sirius red staining. 

Liver enzymes were similarly elevated in both mouse groups. 

Body weight, liver weight, mean hepatic triglyceride content, 

and fasting insulin were similar between groups. Conclusions: 

HIF-1 is necessary for collagen cross-linking in an in vitro model 

of fibrosis. HIF-1 also mediates an increase in liver fibrosis 

in mice with diet induced obesity, even in the absence of IH 

exposure, perhaps due to liver tissue hypoxia in hepatic steatosis. 

This suggests that hepatocyte HIF-1 may play a role in the 

progression of liver fibrosis in patients with NAFLD and OSA, 

and ongoing work will further delineate the precise mechanism 

for this relationship. 

Disclosures: 

The following authors have nothing to disclose: Omar Mesarwi, Mi-Kyung Shin, 

Shannon Bevans-Fonti, Vsevolod Polotsky 

1423 

Treatment of Hepatic Fibrosis using amniotic fluid 

derived mesenchymal stem cells overexpressing interleukin-17 

receptor like molecule 

Peipei Wang 1,2 , Qiyi Zhao 1,2 , Liang Peng 1,2 , Zhanlian Huang 1,2 , 

Shicheng Su 3 , Zhiliang Gao 1,2 ; 1 Department of Infectious Diseases, 

Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 

China; 2 GuangDong Provincial Key Laboratory of Liver Disease, 

Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 

China; 3 Sun Yat-sen University, Guangzhou, China 

Background: Stem cell transplantation has been shown to have 

the capacity to partly reverse liver fibrosis based on its repairing 

and reconstructing effects. However, therapeutic effects of liver 

fibrosis with genetically modified stem cells which have both 

repairing effects and immunotherapy remain uncertain. Our 

study aimed to investigated the therapeutic effects and immunological 

changes in hepatic fibrosis rats after transplantation 

of amniotic fluid derived mesenchymal stem cells over-expressing 

interleukin-17 receptor like molecule (IL-17RLM). Materials 

and methods: Amniotic fluid derived-mesenchymal stem cells 

(AF-MSC) were isolated, and then AF-MSC cell line stably 

over-expressing IL-17RLM (AFMSC-IL-17RLM) was established 

via lentiviral transfection. Liver fibrosis model was established 

in Wista rats after subcutaneous injection of CCl4 twice weekly 

for 9 weeks. AFMSC- IL-17RLM were transplanted twice weekly 

for 3 weeks via tail vein injection. Results: Genetically modified 

stem cells transplantation was safety for liver fibrosis rats; it 

could home to the damaged liver after tail vein injection, with 

no damage to other organs (e.g. brain, lung, spleen). Lower 

hepatic fibrosis score and inflammation grade were observed 

in pathological section of AFMSC- IL-17RLM group, compared 

with the control groups. IL-17, MCP-1, IL-1α, TNF-α showed 

an obvious decrease(p


among the three groups(p=NS).Th17 cells were obviously 

reduced in AFMSC- IL-17RLM group as compared with blank 

groups(p6.5) relative to NAFL or NASH subjects 

without diabetes (ANOVA p=0.056), and correlated with 

HbA1c (r 2 =0.57). FSR of circulating plasma lumican, a proteoglycan 

involved in collagen fibril assembly, correlated with 

liver collagen FSR (r 2 =0.57) and with liver stiffness (r 2 =0.73). 

Conclusion: Hepatic collagen remodeling and synthesis rates 

are higher in NASH than NAFL, increase with fibrosis severity 

and track with diabetes, a risk factor for fibrosis progression. 

Plasma lumican FSR correlates closely with liver collagen FSR 

and liver stiffness. Non-invasive kinetic measurements may be 

helpful as early read outs of treatment response in anti-fibrotic 

trials. The elevated hepatic fibrosis rates in advanced disease 

suggest that hepatic scar is dynamic and potentially reversible.


Disclosures: 

Claire Emson - Employment: KineMed 

Martin Decaris - Employment: KineMed Inc. 

Scott M. Turner - Employment: KineMed, Inc. 

Carine Beysen - Employment: KineMed, Inc; Stock Shareholder: KineMed, Inc 

Kelvin Li - Employment: KineMed, Inc; Stock Shareholder: KineMed, Inc 






The following authors have nothing to disclose: Carolyn Hernandez, Jeffrey Y. 

Cui, Leander Lazaro, David A. Brenner, Marc Hellerstein 

1426 

In vivo cell specific gene silencing in the liver using 

novel siRNA-loaded nanohydrogel particles 

Leonard Kaps; Institute of Translational Immunology and Research 

Center for Immunotherapy (FZI), Mainz, Germany 

Background: Efficient in vivo transport of active siRNA to specific 

liver cells remains challenging. Compared to lipoplex or 

polyplex formulations, cationic nanohydrogel particles (NHP) 

can serve as superior oligonucleotide carriers [Siegwart DJ et 

al, PNAS 2011]. We have designed NHP [Nuhn L et al, ACS 

Nano 2012] that serve as stable carriers for in vivo siRNA 

delivery [Nuhn L et al, Biomacromolecules 2014]. Results: NHP 

composed of block copolymers loaded with negative control 

siRNA up to 600 nM siRNA and of a size of 30 to 40 nm did 

not show cytotoxicity for murine 3T3 fibroblasts, Raw or MHS 

macrophages, HepG2 human liver cells or AML-12 murine 

benign hepatocytes. In full media FITC-labelled NHP were 

taken up efficiently reaching up to 90% after 1 h incubation as 

determined by FACS analysis. NHP loaded with procollagen 

a1(I) or CD68 targeted siRNA yielded a robust knockdown 

in 3T3 and Raw cells, respectively, after 48 h of incubation 

as determined by qPCR. In vivo studies using near infrared 

labelled NHP loaded with Cy5 labelled procollagen a1(I) 

siRNA were performed in lCCl4 treated mice. After i.v. injection 

NHP accumulated in the liver (80%), as determined by in 

vivo and ex vivo near infrared imaging. IHC and ex vivo FACS 

analysis revealed a preferential colocalization with myofibroblasts 

(α-SMA+) and macrophages (CD45+ F4/80+ CD11b+). 

NHP loaded with procollagen a1(I) siRNA generated an up to 

80% in vivo knockdown in CCl4-induced liver fibrosis. Efficient 

knockdown was further confirmed by decreased liver collagen 

(Sirius red staining and hydroxyproline content). Conclusions: 

siRNA loaded NHP accumulate in the liver and especially in 

(myo-) fibroblasts and macrophages/Kupffer cells. Due to their 

intrinsic specificity for nonparenchymal cells, an apparent lack 

of toxicity and a high loading capacity, they are attractive 

carriers for siRNA-based antifibrotic or immune modulatory 

therapies. 

Disclosures: 

The following authors have nothing to disclose: Leonard Kaps 

1427 

Host microbiota dictates the profibrotic impact of PAMPs 

in the liver 

Suriguga Suriguga 1 , Floris Fransen 2 , Dorenda Oosterhuis 1 , Geny 

M. Groothuis 3 , Paul D. Vos 2 , Henricus A. Mutsaers 1 , Peter Olinga 1 ; 

1 Division of Pharmaceutical Technology and Biopharmacy, Department 

of Pharmacy, University of Groningen, Groningen, Netherlands; 

2 Division of Medical Biology, Department of Pathology and 

Medical Biology, University Medical Center Groningen, University 

of Groningen, Groningen, Netherlands; 3 Division of Pharmacokinetics, 

Toxicology and Targeting, Department of Pharmacy, University 

of Groningen, Groningen, Netherlands 

Background Pathogen-associated molecular patterns (PAMPs), 

e.g. lipopolysaccharide (LPS) and flagellin, might promote liver 

fibrosis via the gut-liver axis. However, how these bacterial 

components initiate fibrogenesis needs further clarification. 

In germ-free (GF) mice, the liver has no history of interaction 

with PAMPs, while in colonized (specific pathogen free, SPF) 

mice the liver is exposed to PAMPs. Therefore, this study was 

designed to investigate the possible fibrogenic effect of PAMPs 

in SPF and GF mice, using precision-cut liver slices (PCLS) as 

fibrosis model. Methods PCLS from SPF and GF mice were 

incubated with or without 10 mg/ml LPS or 50 ng/ml flagellin 

for 48h. Viability of PCLS was assessed by measuring their 

ATP content. Gene expression of key fibrosis markers Procollagen1α1 

(Col1α1), α-Smooth Muscle Actin (α-SMA), Heat 

Shock Protein 47 (Hsp47), and Plasminogen Activator Inhibitor-1 

(PAI-1) were determined by real-time qPCR. Results Both 

SPF and GF mice liver slices were viable up to 48h. Solely in 

SPF PCLS, we observed spontaneous onset of fibrosis during 

culture up to 48h. Yet, treatment with LPS or flagellin did not 

augment the fibrotic response. In contrast, in GF PCLS, Hsp47 

expression was significantly up-regulated in the presence of 

LPS: 1.4-fold, while Col1α1 and α-SMA tended to increase 

compared to control slices incubated for 48h. PAI-1, a marker 

for transforming growth factor-β (TGF-β) pathway activation, 

was significantly elevated up to 81.8-fold in SPF and 37.2-fold 

in GF PCLS after culture (48h) compared to directly after slicing. 

Exposure to LPS further increased PAI-1 gene expression in 

GF PCLS (2.9-fold), but not in SPF mice. Moreover, in GF liver 

slices treatment with flagellin significantly increased expression 

of Col1α1 (1.8-fold), α-SMA (1.8-fold), and Hsp47 (1.2-fold), 

while it did not effect PAI-1. Conclusion Both LPS and flagellin 

could exacerbate fibrogenesis in GF mouse liver slices but not


in SPF mouse liver slices. These results indicate that PAMPs are 

profibrotic in the absence of microbiota, whereas in SPF mice 

the microbiota might confer hepatoprotection, which attenuates 

the onset of fibrosis by PAMPs in SPF PCLS. Moreover, LPS 

may rely on the TGF-β signaling pathway to elicit a fibrotic 

response, while flagellin might depend on other pathways. 

Disclosures: 

The following authors have nothing to disclose: Suriguga Suriguga, Floris Fransen, 

Dorenda Oosterhuis, Geny M. Groothuis, Paul D. Vos, Henricus A. Mutsaers, 

Peter Olinga 

1428 

Correlations between hepatic morphometric collagen 

content, histologic fibrosis staging, and serum markers 

in patients with advanced fibrosis due to nonalcoholic 

steatohepatitis (NASH) 

Zachary D. Goodman 1 , Lakshmi Alaparthi 1 , Fanny Monge 1 , Keyur 

Patel 2 , Rohit Loomba 3 , Stephen H. Caldwell 4 , Eric Lawitz 5 , Stephen 

A. Harrison 6 , Mitchell L. Shiffman 7 , Manal F. Abdelmalek 2 , 

Robert P. Myers 8 , Raul E. Aguilar Schall 8 , Mani Subramanian 8 , 

John G. McHutchison 8 , Vlad Ratziu 9 , Arun J. Sanyal 10 , Nezam 

H. Afdhal 11 ; 1 Inova Fairfax Hospital, Falls Church, VA; 2 Duke 

Clinical Research Institute, Durham, NC; 3 University of California 

at San Diego, San Diego, CA; 4 University of Virginia, Charlottesville, 

VA; 5 Texas Liver Institute, University of Texas Health Science 

Center, San Antonio, TX; 6 San Antonio Military Medical Center, 

Fort Sam Houston, TX; 7 Liver Institute of Virginia, Richmond, VA; 

8 Gilead Sciences, Inc., Foster City, CA; 9 Hopital Pitie-Salpretriere, 

Paris, France; 10 Virginia Commonwealth University, Richmond, 

VA; 11 Beth Israel Deaconess Medical Center and Harvard Medical 


Background: Our aim was to determine the relationships 

between hepatic collagen assessed by morphometry with serum 

fibrosis markers, Ishak fibrosis staging, and other histologic 

features of NASH in patients with advanced fibrosis. Methods: 

The study included adults with NASH and bridging fibrosis 

(Ishak stage 3 or 4) or cirrhosis (stage 5 or 6) enrolled in two 

phase 2b trials of simtuzumab, a monoclonal antibody against 

lysyl oxidase-like-2 (LOXL2). Liver biopsies were graded centrally 

according to the NAFLD Activity Score (NAS) and hepatic 

collagen in sirius red-stained biopsies was quantified via computer-assisted 

morphometry. Serum LOXL2 (sLOXL2) was measured 

using an immunoassay (VIDAS® LOXL2; bioMérieux, 

Marcy L’Etoile, France). The associations between hepatic collagen 

and Ishak fibrosis stage, noninvasive fibrosis markers 

(sLOXL2, FibroTest, ELF, APRI, FIB-4, and NAFLD Fibrosis Score 

[NFS]), and the NAS and its components were determined. 

Results: 429 of 477 randomized patients (89.9%) with biopsies 

acceptable for computerized morphometry were included. 

The median age was 56 years (IQR 50-60), 62% were female, 

52% had cirrhosis, and the median hepatic collagen content 

was 8.4% (IQR 5.2-14.7%). Hepatic collagen was moderately 

correlated with fibrosis stage (Spearman ρ=0.59; P


1429 

Concerted effects of the two major genetic risk factors 

for cholestatic (ABCB4) and fatty liver disease (PNPLA3) 

on liver fibrosis: elastography-based study in chronic 

liver disease 

Marcin Krawczyk 2,1 , Frank Grünhage 2 , Frank Lammert 2 ; 1 Laboratory 

of Metabolic Liver Diseases, Department of General, Transplantation 

and Liver Surgery, Medical University of Warsaw, 

Warsaw, Poland; 2 Department of Medicine II, Saarland University 

Medical Center, Homburg, Germany 

Background: The liver responds uniformly with liver fibrosis 

to any chronic injury. Genome-wide association studies 

have established the common polymorphism p.I148M of the 

PNPLA3 gene as the major genetic determinant of non-alcoholic 

and alcoholic fatty liver disease. Whereas rare mutations 

of the hepatobiliary phosphatidylcholine transporter ABCB4 

are known to cause biliary cirrhosis, large-scale whole-genome 

sequencing lead to the identification of the common ABCB4 

variant c.711A>T as general risk factor for cholestasis and cirrhosis 

(Nat Genet 2015). Hence, our hypothesis now was that 

the combination of these two most relevant genetic risk factors 

should aggravate fibrosis across chronic liver diseases (CLD). 

Patients and methods: To address this issue, we studied 713 

patients (age 50±12 years, 434 men) recruited for our non-invasive 

elastography-based study (J Hepatol 2011), which confirmed 

that the PNPLA3 mutation represents a common fibrosis 

risk gene. Liver stiffness was determined non-invasively using 

transient elastography (TE, Fibroscan). The PNPLA3 c.444C>G 

(p.I148M) and ABCB4 c.711A>T (splice region variant) polymorphisms 

were genotyped by PCR-based assays. Association 

with transient elastography results was tested in contingency 

tables, and TE values in carriers of distinct PNPLA3 and ABCB4 

genotypes were compared by non-parametric tests. Results: 

The ABCB4 c.711 ([TT] n = 23, [AT] n = 204, [AA] n = 486) 

and the PNPLA3 p.I148M ([CC] = 382, [CG] = 280 and [GG] 

= 50) genotype distributions were in Hardy-Weinberg equilibrium. 

The median liver stiffness in the entire cohort was 6.7 

kPa and ranged from 2.2 to 75.0 kPa; 156 patients presented 

with TE ≥ 13.0 kPa. Of note, the procholestatic allele ABCB4 

c.711A was present in the majority of the patients and did not 

significantly influence TE per se. The combination of ABCB4 

and PNPLA3 risk alleles lead to incremental increases of mean 

TE from 5.6 kPa in carriers of 0 to 7.8 kPa in carriers of 4 risk 

alleles. In heterozygous carriers of the PNPLA3 risk variant 

p.I148M, the presence of the procholestatic ABCB4 geneotype 

[AA] was significantly (P = 0.04) associated with increased 

TE; in contrast, this effect was absent in PNPLA3 homozygotes. 

Conclusions: This is the first study to assess simultaneously the 

effects of the currently known most common mutations in two 

distinct core pathways simultaneously in patients with CLD. 

Our results suggest that the procholestatic ABCB4 variant 

might boost the harmful effects of the major PNPLA3 mutation 

and vice versa. Prospective studies are warranted to evaluate 

ABCB4-PNPLA3 compound genotypes as genetic determinants 

of liver vulnerability. 

Disclosures: 

Frank Grünhage - Grant/Research Support: Gilead; Speaking and Teaching: 

Falk, Abbvie 

The following authors have nothing to disclose: Marcin Krawczyk, Frank Lammert 

1430 

Innate Lymphoid Cell (ILC) Subset Composition of 

Human Liver: Enrichment for ILC1s in Hepatic Cirrhosis 

Lucy Golden-Mason 1 , Silvia Giugliano 1 , Linling Cheng 1 , Hugo R. 

Rosen 1,2 ; 1 GI/Hepatology, Univ Colorado Denver, Aurora, CO; 

2 Veteran’s Affairs Medical Center, Denver, CO 

Introduction: Innate lymphoid cells (ILCs) are emerging as 

important immune effectors that, in addition to stimulating proand 

anti-inflammatory responses, are involved in tissue remodeling. 

Three groups of ILCs have been identified in humans. 

These include pro-inflammatory Group 1 (ILC1) populations 

which produce IFN-γ and ILC3s which produce IL-17 and or 

IL-22. ILC2s are characterized by the production of anti-inflammatory 

cytokines, predominantly IL-13. In an acute liver injury 

model, natural cytotoxicity receptor (NCR) negative ILC3s are 

tissue protective. The ILC composition of human liver is unexplored. 

We hypothesized that the human liver, an established 

site for innate immunity, would harbor pro-inflammatory ILCs 

(ILC1/ILC3) which would be increased in hepatic inflammation 

and cirrhosis. Methods: Single cell suspensions were prepared 

from liver tissues using mechanical disruption and enzymatic 

digestion. Our study group consisted of normal (n=12), alcoholic 

liver disease (ALD, n=6), cholestatic liver disease (PBC/ 

PSC, n=6) and HCV (n=18). The majority of subjects (88%) 

were Caucasian and 68% were male. Multi-parameter flow 

cytometric analysis was used to characterize human hepatic 

ILCs. Total ILCs were identified by their lymphoid morphology, 

absence of markers for myeloid, T, NK, B and stem cells 

and by co-expression of CD45 and the IL-7 receptor (CD127). 

ILC subsets were identified by the expression pattern of c-kit 

(CD117), NCR NKp44 and CRTH2. Results: In normal liver 

ILCs represent 4.45% (1.39-9.1) of CD45-positive lineage 

negative lymphoid cells. ILC1 are the predominant subset 

detected in normal liver followed by NCR neg ILC3s. Anti-inflammatory 

ILC2s comprise 2.39-16.8% of total ILCs in normal 

liver. In cirrhotic liver tissues levels of total ILCs are stable in 

HCV-induced and cholestatic liver disease-related cirrhosis. 

However, in alcohol-induced cirrhosis a greater than 3-fold 

increase in ILC levels was observed (p


1431 

Risk of cirrhosis in first degree relatives of patients with 

NAFLD cirrhosis: A prospective study 

Rohit Loomba 1 , Jeffrey Y. Cui 2 , Ricki Bettencourt 3 , Nicholas 

Schork 4 , Chi-Hua Chen 5 , Mahdi Al Ikhwan 2 , Archana Bhatt 2 , 

Lisa M. Richards 2 , Claude B. Sirlin 5 , David A. Brenner 2 ; 1 Division 

of Gastroenterology and Epidemiology, University of California 

at San Diego, La Jolla, CA; 2 Gastroenterology, UCSD School of 

Medicine, La Jolla, CA; 3 Family and Preventive Medicine, UCSD 

School of Medicine, La Jolla, CA; 4 J Craig Venter Institute, La Jolla, 

CA; 5 Radiology, UCSD School of Medicine, La Jolla, CA 

Background: The risk of cirrhosis in the first-degree relative of 

patients (pts) with NAFLD-cirrhosis is unknown & needs to be 

determined. This determination would help develop guidelines 

for cirrhosis screening in first-degree relatives of patients with 

NAFLD-cirrhosis. Aim: To determine the risk of NAFLD-cirrhosis 

in first-degree relatives of pts with known NAFLD cirrhosis. 

Methods: This is a cross-sectional analysis of two prospectively 

recruited familial cohorts: the familial NAFLD-cirrhosis cohort & 

the control-families cohort. The familial cirrhosis cohort included 

pts with known NAFLD-cirrhosis (probands) recruited from the 

UCSD NAFLD Clinic & their first-degree relatives. The controls 

were derived from a community-dwelling cohort of normal 

twin/sib-sib/parent-offspring pair with one random participant 

as a control & the respective pair as the first-degree relative 

recruited from the general population of Southern California. 

Probands had documented evidence of NAFLD (based upon 

AASLD Guidelines) & either biopsy-proven or imaging-proven 

cirrhosis. The first-degree relatives of pts with NAFLD-cirrhosis 

& controls underwent a research visit & advanced MRI +MRE 

to detect advanced fibrosis (MRE liver stiffness >3.63 kPa has 

an AUROC 0f 0.93). Multivariable-adjusted logistic regression 

(odds ratio [OR]) was done to examine the risk of cirrhosis in 

first-degree relatives of pts with cirrhosis. Results: This study 

included 19 probands (mean age 62.5 yrs & BMI 32.3 kg/ 

m 2 ) with NAFLD-cirrhosis, & 36 first-degree relatives (mean 

age 47.4 yrs & BMI 30.9 kg/m 2 ) of probands. The controls 

(mean age 40.9 yrs & BMI 25.5 kg/m 2 ) included 61 pairs 

(total n=122) of either twin/sib-sib/parent-offspring pair; 

with 61 without any evidence of NAFLD or cirrhosis, and 61 

first-degree relatives of these randomly ascertained unaffected 

first-degree relative pairs. The prevalence of NAFLD-cirrhosis in 

the first degree relatives of pts with NAFLD-cirrhosis was significantly 

higher than the controls (13.9 % vs. 1.6%, p


1433 

Safety, Pharmacokinetics, and Biologic Activity of 

ND-L02-s0201, a Novel Targeted Lipid-Nanoparticle to 

Deliver HSP47 siRNA for the Treatment of Patients with 

Advanced Liver Fibrosis: Interim Results from Clinical 

Phase 1b/2 Studies 

Eric Lawitz 2 , Yasunobu Tanaka 3 , Fred Poordad 2 , Julio A. Gutierrez 

2 , Kathy Carr 4 , Wenbin Ying 1 , Yoshiro Niitsu 5 , Kageshi 

Maruyama 3 ; 1 NDT, Oceanside, CA; 2 Texas Liver Institute, University 

of Texas Health Science Center, San Antonio, TX; 3 Nitto Denko 

Corporation, Tokyo, Japan; 4 RRD International, RRD International, 

LLC, Rockville, MD; 5 Sapporo Medical University, Sapporo, Japan 

Objectives: Therapy for liver fibrosis is a major unmet medical 

need. We have reported a novel therapeutic modality that 

inhibits expression of the collagen-specific chaperone, heat 

shock protein 47 (HSP47), with stellate cell (SC)-specific drug 

delivery (Sato et al. 2008 Nature Biotech), with a goal of 

reversal of collagen deposition. NDL02s0201 is an injectable 

lipid nanoparticle formulation with a novel vitamin A analogue 

targeting agent for SC and a chemically modified small interfering 

ribonucleic acid (siRNA) active ingredient that inhibits 

HSP47. We report safety, PK and preliminary biologic activity 

results from a Phase 1a study and an ongoing Phase 1b/2 

study. Methods: The Phase 1a study was a randomized, double-blind, 

placebo-controlled study to evaluate the safety and 

PK of escalating single doses of ND-L02-s0201 from 0.03 to 

0.8 mg/kg in normal volunteers. The Phase 1b/2 clinical trial 

is an ongoing, open label, multiple-dose, escalation study to 

evaluate safety, PK, and biological activity in patients with 

METAVIR F3-4. NDL02s0201 is intravenously infused once or 

twice weekly for 5 weeks at 0.2 to 0.6 mg/kg/week. Results: 

The Phase 1a study included 56 subjects (42 ND-L02-s0201, 

14 placebos). Seven subjects treated with ND-L02-s0201 at 

dose levels of 0.03 to 0.6 mg/kg had treatment-related mild/ 

moderate adverse events characteristic of hypersensitivity infusion 

reactions. After premedication with H1 and H2 blockers 

and a slower infusion rate, no infusion reactions occurred 

at 0.8 mg/kg. PK results of the siRNA were linear over the 

range of doses studied. The mean elimination half-life across 

the 7 doses was 27.3 h with no apparent dose-related trends. 

Available data from the Phase 1b/2 study includes 8 subjects 

from the 1 st dose cohort: 4 with NASH and 4 with HCV. The 

baseline (within 12 months before enroll) METAVIR score was 

F3 for 3 subjects and F4 for 5 subjects. Cohort 1 (0.2 mg/kg/ 

week) of the Phase 1b/2 study was complete. Two possibly 

treatment-related gastrointestinal AEs were reported (epigastric 

discomfort and abdominal pain). At the lowest planned dose 6 

out of 8 subjects had at least 1 stage reduction in Ishak score 

(mean reduction 1.25, range 0-4, N=8), and 1 subject showed 

reduction of METAVIR score from F-3 to F-0 right after 5 weeks 

dosing. Conclusion: ND-L02-s0201 was well tolerated in both 

studies with no dose limiting toxicities up to 0.8 mg/kg (single) 

and of 0.2 mg/kg/week (multiple doses). The PK results were 

consistent between healthy subjects and patients as well as for 

single and multiple doses with no drug accumulation. Histologic 

improvement was seen in the majority of patients at the 

lowest dose studied. 

Disclosures: 













Wenbin Ying - Management Position: Nitto Denko Technical Corporation 

The following authors have nothing to disclose: Yasunobu Tanaka, Julio A. Gutierrez, 

Kathy Carr, Yoshiro Niitsu, Kageshi Maruyama 

1434 

Antifibrotic efficacy of a TGF-β kinase inhibitor on 

early-onset and end-stage of fibrosis in precision-cut 

human liver slices 

Theerut Luangmonkong 1,2 , Suriguga Suriguga 1 , Emilia Bigaeva 1 , 

Dorenda Oosterhuis 1 , Koert P. de Jong 3 , Detlef Schuppan 4,5 , Henricus 

A. Mutsaers 1 , Peter Olinga 1 ; 1 Pharmaceutical Technology 

and Biopharmacy, University of Groningen, Groningen, Netherlands; 

2 Pharmacy, Mahidol University, Bangkok, Thailand; 3 Hepato-Pancreato-Biliary 

Surgery and Liver Transplantation, University 

Medical Center Groningen, Groningen, Netherlands; 4 Medicine, 

Translational Immunology and Research Center for Immunotherapy, 

University of Mainz Medical Center, Mainz, Germany; 5 Gastroenterology, 

Beth Israel Deaconess Medical Center, Harvard 

Medical School, Boston, MA 

Background Advanced liver fibrosis is a major cause of liver 

related morbidity and mortality, but to date, preclinically effective 

antifibrotic compounds have failed to show efficacy in 

clinical trials. Galunisertib, a TGF-β receptor kinase inhibitor, is 

currently studied for the treatment of cancers, including hepatocellular 

carcinoma. Due to its mode of action, galunisertib is 

also a potential candidate for the treatment of liver fibrosis. Precision-cut 

liver slices (PCLS) are an ideal human model to test 

the antifibrotic efficacy of compounds. Therefore, we aimed 

to assess the antifibrotic potency of galunisertib in both the 

early-onset and end-stage of fibrosis in human PCLS. Methods 

Early-onset fibrosis was studied in PCLS prepared from surgical 

excess material of donor livers for reduced-size liver transplantations. 

End-stage fibrosis was studied in PCLS from explanted 

cirrhotic livers of patients undergoing liver transplantation. 

PCLS were incubated with galunisertib (0.625-10 μM) for 

48h. Viability was assessed by ATP content of the slices, and 

gene expression of the key fibrosis markers Procollagenα1(I) 

(COL1α1), α-smooth muscle actin (α-SMA), Heat shock protein 

47 (HSP47), and Plasminogen activator inhibitor-1 (PAI-1), 

was assessed by real-time qPCR. Results Liver slices remained 

viable for 48h. In the early-onset of fibrosis, COL1α1 gene 

expression increased significantly 3.5-fold, while expression 

of α-SMA, HSP47, and PAI-1 remained unaltered. Following 

treatment with galunisertib, only the highest concentration lowered 

the ATP content by about 20 %, while the fibrosis markers 

decreased in a concentration-dependent manner up to 82 ± 3 

%, 55 ± 7 %, and 71 ± 4 %, for COL1α1, HSP47, and PAI-1, 

respectively. α-SMA gene expression did not show a concentration-dependent 

inhibition, and only the highest concentration 

of galunisertib significantly decreased gene expression by 22 

± 9 %. In the cirrhotic PCLS, COL1α1 gene expression significantly 

increased 6.5-fold after 48h of incubation, while the 

other genes remained unchanged. The highest concentration 

of galunisertib in the early-onset of fibrosis study (10 μM) significantly 

inhibited the expression of COL1α1 by 81 ± 11 % 

and PAI-1 by 78 ± 4 % in cirrhotic PCLS, without toxicity, while 

HSP47 and α-SMA expression remained unchanged. Conclusion 

Human PCLS that reflect early and late stage fibrosis, 

appear to be a viable tool to predict antifibrotic effects prior to 

clinical studies. Galunisertib is an attractive potential drug for 

the treatment of human liver fibrosis.


Disclosures: 

The following authors have nothing to disclose: Theerut Luangmonkong, Suriguga 

Suriguga, Emilia Bigaeva, Dorenda Oosterhuis, Koert P. de Jong, Detlef Schuppan, 

Henricus A. Mutsaers, Peter Olinga 

1435 

Serum lysyl oxidase-like-2 (sLOXL2) levels correlate with 

fibrosis stage in patients with nonalcoholic steatohepatitis 

(NASH) 

Stephen A. Harrison 1 , Zachary D. Goodman 2 , Vlad Ratziu 3 , Rohit 

Loomba 4 , Anna Mae Diehl 5 , Eric Lawitz 6 , Holger Hinrichsen 7 , 

Kiran Bambha 8 , Manal F. Abdelmalek 5 , Robert P. Myers 9 , Raul 

E. Aguilar Schall 9 , Mani Subramanian 9 , John G. McHutchison 9 , 

Nezam H. Afdhal 10 , Andrew J. Muir 5 ; 1 San Antonio Military 

Medical Center, Fort Sam Houston, TX; 2 Inova Fairfax Hospital, 

Falls Church, VA; 3 Hôpital Pitié-Salpêtrière, Paris, France; 4 University 

of California at San Diego, San Diego, CA; 5 Duke Clinical 

Research Institute, Durham, NC; 6 Texas Liver Institute, University 

of Texas Health Science Center, San Antonio, TX; 7 Gastroenterologisch-Hepatologisches 

Zentrum, Kiel, Germany; 8 University of 

Colorado Denver, Aurora, CO; 9 Gilead Sciences, Inc., Foster City, 

CA; 10 Beth Israel Deaconess Medical Center and Harvard Medical 


Background: LOXL2 plays a central role in liver fibrosis by 

catalyzing collagen cross-linkage. Our aim was to examine 

the association between sLOXL2 and disease severity in 

patients with advanced fibrosis due to NASH. Methods: The 

study included adults with NASH and bridging fibrosis (Ishak 

stage 3 or 4) or cirrhosis (stage 5 or 6) enrolled in two phase 

2b trials of simtuzumab, a monoclonal antibody against lysyl 

oxidase-like-2 (LOXL2). sLOXL2 was measured using an immunoassay 

(VIDAS® LOXL2; bioMérieux, Marcy L’Etoile, France) 

in NASH patients and 50 healthy controls. Liver biopsies were 

graded centrally according to the NAFLD Activity Score (NAS) 

and fibrosis was staged according to the Ishak classification. 

The associations between sLOXL2 and fibrosis stage, NAS, 

demographics, body mass index (BMI), diabetes, liver biochemistry, 

MELD, and fibrosis markers (FibroTest, ELF, APRI, 

FIB-4, NAFLD Fibrosis Score [NFS]) were determined. Results: 

474 of 477 randomized patients (99.4%) with available 

sLOXL2 and stage 3-6 fibrosis were included. The median age 

was 56 years (IQR 50-60), 63% were female, 68% had diabetes, 

the median BMI was 33.6 kg/m 2 (IQR 29.8-38.2), and the 

median serum ALT was 39 U/L (IQR 28-62). Median sLOXL2 

was higher in patients with NASH than controls (107 pg/mL 

[IQR 82-147] vs. 81 pg/mL [IQR 71-108]; P


no change after treatment. All patients were divided equally 

into training and testing cohorts. The fibrosis scoring index for 

CHC was analyzed against Ishak staging system as the gold 

standard. Results: 27 out of 100 parameters showed high-level 

correlation with Ishak staging, mainly in the portal and total 

collagen compartments. Systematic area under curve (AUC) 

optimization analysis further identified 6 parameters with the 

highest correlation out of the 27. These 6 parameters were as 

efficient (AUC: 0.86–0.94) in identifying differences between 

all Ishak brosis stages as using all 27 parameters (AUC: 0.85– 

0.94). Systematic analysis performed for the 39 paired biopsies 

identified 8, 1 and 2 parameters for the respective Ishak 

staging decrease, increase, and no change groups. These 11 

parameters from the paired biopsies comprised 7 which were 

already identified in the 27 parameters, and 4 new ones. Taking 

the best correlating parameter from these 4 new ones with 

Ishak staging, and adding it to the 6 highest correlation parameters, 

we developed a 7-parameter indexing model (comprising 

Strlength, Portal, NoShortStrP, NoStrPA, NoLongStrPA, 

StrLengthPA, NoXlinkP). This model can faithfully and reliably 

recapitulate Ishak brosis scores in CHC patients, including the 

paired pre- and post-treatment biopsies. Conclusions: A modified 

qFibrosis model was developed and validated for CHC 

patients and for treatment response assessment. An indexing 

system with 7 parameters was shown to be highly accurate 

and reproducible for the objective and quantitative analysis of 

fibrosis progression and/or regression in CHC patients. 

Disclosures: 

Lai Wei - Advisory Committees or Review Panels: Gilead, AbbVie; Grant/ 


The following authors have nothing to disclose: Huiying Rao, Feng Liu, Aileen -. 

Wee, Bo Feng, Ming Yang 

1437 

Collagen and tissue turnover as function of age – implications 

for fibrosis 

Morten A. Karsdal 1 , Federica Genovese 1 , Emilie A. Madsen 1 , 

Tina Manon-Jensen 1 , Detlef Schuppan 2,3 ; 1 Nordic Bioscience, Herlev, 

Denmark; 2 Institute of Translational Immunology and Research 

Center for Immunotherapy, University Medical Center, Mainz, 

Germany; 3 Division of Gastroenterology, Beth Israel Deaconess 

Medical Center, Harvard Medical School, Boston, MA 

Background: The extracellular matrix (ECM) is the backbone of 

all tissues. It is a complex grid consisting of multiple structural 

proteins which each play a vital role for the function and maintenance 

of normal tissue function. In development and growth, 

tissue is being formed and elaborated (tissue modeling), while 

in adult life, tissues are being maintained and remodeled. 

These processes involve likely different mechanisms. During 

tissue modeling and remodeling, small fragments of proteins 

are released into the circulation, where they may be used as 

biomarkers of tissue turnover. Aim: The aim of the study was 

to investigate ECM turnover in rodents as a function of age. 

Materials and Methods: Serum of rats of 1, 2, 3, 4, 5, 6, 

10 and 12 months of age was profiled for 15 markers of 

ECM turnover, including: fragments of type I, II, III, IV, V and 

VI collagen formation (P1NP, Pro-C3, P4NP_7S, Pro-C5, Pro- 

C6) and degradation (C1M, C2M, C2M-beta, C3M, C4M, 

C5M, C6M); biglycan (BGM) and elastin (ELM7) degradation; 

and the type I and II collagen telopeptides CTX-I and CTX-II. 

Results: Type I and II collagen turnover was up to 93% and 

97% downregulated in old (one year) compared to young (one 

month) old animals (p


1439 

Comparison of 16 blood and/or elastometric fibrosis 

tests in 5 causes of chronic liver diseases: too much? 

Towards a simplification 

Jerome Boursier 1,2 , Alexandra Ducancelle 3,2 , Vincent Leroy 4 , Julien 

Vergniol 5 , Nathalie Sturm 6 , Brigitte Le Bail 7 , Jean-Pierre H. Zarski 4 , 

Victor de Ledinghen 5 ; 1 Hepato-Gastroenterology Departement, 

University Hospital, Angers, France; 2 HIFIH Laboratory EA3859, 

University, Angers, France; 3 Virology Department, University Hospital, 

Angers, France; 4 Hepato-Gastroenterology Department, 

University Hospital, Grenoble, France; 5 Hepatology Department, 

University Hospital, Bordeaux, France; 6 Pathology Department, 

University Hospital, Grenoble, France; 7 Pathology Department, 

University Hospital, Bordeaux, France 

Introduction: Most non-invasive fibrosis tests were developed 

in chronic hepatitis C (CHC), but are frequently used in other 

causes. A lot of tests are available. Therefore, we compared 

the accuracy of several usual tests between main etiologies to 

try to determine whether a test could be the most performant 

in several etiologies. Methods: Populations included 1660 

patients: 698 with CHC, 178 with hepatitis B (CHB), 444 

with HIV/CHC, 225 with NAFLD, and 115 with alcoholic liver 

disease (ALD). 16 tests (13 blood tests, 1 elastometry by VCTE 

-Fibroscan- and 2 combining blood markers and elastometry) 

were evaluated. Reference was Metavir fibrosis (F) stage by 

liver biopsy. Results: Accuracy was sensitive to biopsy length. 

Obuchowski indices, reflecting accuracy for all F stages, were 

by decreasing order, CHC: Elasto-FibroMeter VCTE2G/3G : 0.812, 

FibroMeter VIRUS2G : 0.797, FibroMeter VIRUS3G : 0.785, CirrhoMeter 

VIRUS2G : 0.771, Fibrotest: 0.762, CirrhoMeter VIRUS3G : 

0.756, Fibroscan: 0.754, Hepascore: 0.752, FibroMeter ALD : 

0.750, APRI: 0.742, FIB4: 0.741. CHB: Obuchowski indices 

were roughly the same as in CHC. HIV/CHC: Obuchowski 

indices were moderately decreased compared to CHC except 

for Fibrotest (p=0.03). NAFLD: 7 tests had a substantial accuracy 

loss compared to CHC, especially Fibrotest (p=0.006), 

while 5 tests had a substantial gain especially FibroMeter ALD 

(p=0.039) and Zeng score (p=0.030). ALD: 2 tests had a 

dramatic accuracy loss compared to CHC: APRI (p


1441 

Comparing the liver stiffness by transient elastography 

in cirrhosis patients with PBC and HBV to predict the 

risk of esophageal variceal bleeding 

Ying Huang 1 , Haiyun Zhang 1 , Li Xiao 1 , Liqing Zhu 1 , Qingjing 

Zhu 2 , Xiaohua Hou 1 , Ling Yang 1 ; 1 Division of Gastroenterology,Department 

of Internal Medicine, Huazhong University of Science&Technology, 

Wuhan, China; 2 Wuhan Medical Treatment 

Center, Wuhan, China 

Background&Aim: Staging hepatic fibrosis is critical for the 

evaluation and deternination of prognosis in chronic liver diseases. 

Variceal bleeding is the servere complications of liver 

cirrhosis. Endosocopy as the only means to evaluate the varices 

and risk of bleeding is not an easy examination to be 

accepted by patients with expensive price and poor tolerance. 

Transient elastography (TE) is useful as non-invasive diagnositic 

method for predicting liver stiffness and portal hypertension. 

Our aim was to analyze the diagnostic accuracy of liver stiffness 

measurement(LSM) and the esophageal variceal grading 

and the risk of esophageal variceal bleeding in different etiological 

cirrhosis patients with HBV infection or primary billiary 

cirrhosis by transient elastography. Patients and Methods: 

Endoscopy and transient elastography were performed in 24 

primary biliary cirrhosis patients and 66 hepatitis B related 

cirrhosis. The relationships of the liver stiffness between esophageal 

variceal degree and the risk of esophageal variceal 

bleeding, as well as the hematological and biochemical laboratory 

parameters were analyzed. Results: The grade of 

esophageal varices is highly correlated with the stiffness of 

the liver(P


Disclosures: 





Christophe Hézode - Speaking and Teaching: Roche, BMS, MSD, Janssen 




Steven L. Flamm - Advisory Committees or Review Panels: Gilead, Bristol Myers 

Squibb, AbbVie, Janssen, Salix; Consulting: Merck, Janssen, Bristol Myers 

Squibb, AbbVie, Salix, Gilead; Grant/Research Support: Janssen, Bristol Myers 

Squibb, Gilead, AbbVie; Speaking and Teaching: Salix 





Sciences, Inc 



Sciences 



Sciences 








Squibb 

1443 

How gold is the “gold standard” method for staging 

liver fibrosis? 

Ioan Sporea; University of Medicine and Pharmacy Timisoara, 

Timisoara, Romania 

Backgrounds and aim: Liver biopsy (LB) is considered the gold 

standard method for staging liver fibrosis. The aim of our study 

was to assess the quality of liver samples obtained by percutaneous 

LB in a large cohort and to determine the sensitivity of 

this method for staging liver fibrosis. Material and method: We 

performed a retrospective study on echo-assisted LB performed 

in our Department, using 1.4 mm and 1.6 mm Menghini modified 

needles, with 2 liver passages. 1012 LBs performed were 

analyzed (41.4% male, 58.5% female, mean age of 46±12 

years). Based on the length of the liver specimens obtained we 

divided the LBs into 4 groups (< 15 mm; 15- 24 mm; 25-39 

mm; > 40 mm). We calculated the length means for every 

group and usingBedossa’s (1) study we analyzed the percentage 

of expectedcorrectly classified biopsies according to the 

length of biopsy specimen. Results: The overall mean length of 

liver specimen obtained in our cohort was 33±9 mm in size, 

with mean number of portal tracts of 20±10. 1%(10) of the LBS 

were included in the first group (< 15 mm) with mean length of 

98±2 mm. 13% (135) LBS were included in the second group 

(15- 24 mm) obtaining a mean length of 20±1.8 mm.41% 

(418) of the LBs had between 25 and 39 mm with a mean 

length of 30±3 mm. 45% (449) of the LBs obtained specimens 

larger than 40 mm with mean length of 42±5 mm.Using 

Bedossa’s study (1) and diagram considering the sensitivity of 

LB for staging liver fibrosis according to the length of biopsy 

specimen we obtained the following sensitivities: Group 1 (< 

15 mm) 55%; Group 2 (15-24 mm) 70%; Group 3 (25-39 

mm) 75%; Group 4 (> 40 mm) 83% and an overall sensitivity 

of 80%. Conclusions:Despite the fact that good liver specimens 

are obtained using Menghini needles with 2 passages technique 

the overall sensitivity of liver biopsy is only around 80% 

using Bedossa criteria. References: 1. Bedossa P et al. Sampling 

variability of liver fibrosis in chronic hepatitis C. Hepatology 

2003;38:1449–1457. 

Disclosures: 

The following authors have nothing to disclose: Ioan Sporea 

1444 

Reproductive Aging, Independent of Chronologic Age, 

Is Associated with Accelerated Fibrosis Progression in 

HIV/HCV Co-Infected Women 

Monika Sarkar 1 , Jennifer L. Dodge 1 , Ruth Greenblatt 1 , Mark H. 

Kuniholm 2 , Jack Dehovitz 3 , Michael Plankey 9 , Andrea Kovacs 4 , 

Audrey French 7 , Eric C. Seaberg 8 , Igho Ofotokun 5 , Margaret Fischl 

6 , Edgar T. Overton 10 , Erin Kelly 1 , Peter Bacchetti 1 , Marion G. 

Peters 1 ; 1 University of California, San Francisco, San Francisco, 

CA; 2 Albert Einstein College of Medicine, Bronx, NY; 3 SUNY 

Downstate Medical Center, Brooklyn, NY; 4 University of Southern 

California, Alhambra, CA; 5 Emory University, Atlanta, GA; 6 University 

of Miami, Miami, FL; 7 Cook County Health and Hospitals 

System, Chicago, IL; 8 Johns Hopkins University, Baltimore, MD; 

9 Georgetown University Medical Center, Washington, DC; 10 University 

of Alabama, Birmingham, AL 

Cross sectional studies note less fibrosis in pre- versus post 

menopausal women, though data lack serial fibrosis measures 

and adequate adjustment for chronologic aging. Methods: 

This is a retrospective cohort study of HIV/HCV co-infected 

women, followed in the multicenter Women’s Interagency HIV 

Study. Eligible women were pre-menopausal at entry (n=411), 

confirmed by detectable anti-mullerian hormone (AMH). Subsequent 

AMH loss during follow-up indicated menopausal 

transition. We assessed fibrosis rate using the validated fibrosis 

marker, FIB4 (level < 1.45 indicates < stage 3 fibrosis). 

Fibrosis rate was evaluated in each woman as she transitioned 

through menopause using random-intercept-random-slopes, 

accounting for chronologic age with flexible linear splines. 

Reproductive age was defined as pre- (detectable AMH), peri- 

(≤ 5 yrs after AMH loss) and post-menopause (> 5 yrs after 

AMH loss). Covariates included race, metabolic factors, alcohol, 

and antiretroviral therapy (ART). Results: Median follow-up 

was 11.5 (IQR 6-18.5) years with mean age of 44.5 (+/- 7.0) 

and median entry FIB4 of 1.71 (IQR 1.2-2.6). Adjusted for 

chronologic aging, fibrosis was 0.126 FIB4 per year faster 

(95% CI 0.029-0.224, p=0.01) during peri-menopause and 

0.149 FIB4 per year faster (95% CI 0.003-0.295, p=0.046) 

during post- as compared to pre-menopause. After adjusting 

for race, alcohol, and ART, accelerated fibrosis persisted in 

peri- compared to pre-menopause (0.103 FIB4/year faster, 

95% CI 0.007-0.200, p=0.04), though not significant (0.117, 

95% CI -0.029-0.0262, p=0.12) for post- vs pre-menopausal 

comparison. Conclusion: In HIV/HCV co-infected women, liver 

fibrosis accelerates through peri-menopause, independent of 

chronologic aging. Peri-menopausal women may have more 

urgent need for HCV therapy. Longitudinal analyses of hepatic 

fibrosis across reproductive age in women with non HCV-related 

liver disease are warranted.


when AZA 1nΜ was applied in SFM as well as in TGF induced 

HSCs: For COL1A1: SFM 1.45 vs AZA 0.47 (p


1447 

A novel canine liver cirrhosis model to develop less 

invasive liver regeneration therapy using cultured bone 

marrow-derived cells 

Takashi Matsuda 1,2 , Taro Takami 1 , Yuki Aibe 1 , Toshihiko Matsumoto 

1,4 , Tsuyoshi Ishikawa 1 , Naoki Yamamoto 1,5 , Shuji Terai 3 , 

Isao Sakaida 1,6 ; 1 Department of Gastroenterology and Hepatology, 

Yamaguchi University Graduate School of Medicine, Ube, 

Yamaguchi, Japan; 2 Sanyo-Onoda General Hospital, Sanyo-Onoda, 

Japan; 3 Division of Gastroenterology and Hepatology, 

Niigata University, Niigata, Japan; 4 Department of Oncology and 

Laboratory Medicine, Yamaguchi University Graduate School of 

Medicine, Ube, Yamaguchi, Japan; 5 Yamaguchi University Health 

Administration Center, Yamaguchi University, Ube, Yamaguchi, 

Japan; 6 Center for Reparative Medicine, Yamaguchi University 

Graduate School of Medicine, Ube, Yamaguchi, Japan 

Background/Aims: We developed autologous bone marrow 

cell infusion (ABMi) therapy for decompensated liver cirrhosis 

patients using non-cultured autologous whole bone marrow 

(BM) cells aspirated under general anesthesia, and then 

reported the safety and efficacy of this approach. We have 

also been developing a new, less invasive liver regeneration 

therapy using cultured autologous BM-derived mesenchymal 

stem cells (BMSCs) from small amounts of BM fluid aspirated 

under local anesthesia. Before human clinical trials, the safety 

and efficacy of cultured autologous BMSC infusion in mediumto-large 

animals must be confirmed; thus, we aimed to develop 

a canine liver cirrhosis model. Methods: Eight 1- to 2-year-old 

beagles were studied. A small amount of BM fluid was aspirated 

from the canine humerus to assess the characteristics of 

BMSCs. We implanted a human peripherally inserted central 

venous catheter (PICC) in the stomach and a subcutaneous 

infusion port in the back of the neck of each canine. Repeated 

injection of carbon tetrachloride (CCl 4 

) through the implanted 

catheter (high-dose period: 0.75 mL/kg body weight, twice a 

week for 6 weeks; low-dose period: 0.20 mL/kg body weight, 

twice a week for 4 weeks) was performed to induce liver cirrhosis. 

After 10 weeks of CCl 4 

injection, eight canines were 

equally divided into two groups: no cell infusion (control group) 

and autologous BMSC infusion via the peripheral vein (BMSC 

group). Low-dose CCl 4 

was continued after BMSC infusion, and 

blood examinations, ultrasonography-guided liver biopsies, 

and indocyanine green (ICG) tests were carried out before and 

at 4 weeks after the infusion. Results: Canine BMSCs cultured 

in standard DMEM with 10% FBS adhered to plastic and were 

CD44+, CD90+, and CD45−. They differentiated into adipocytes 

and osteocytes, consistent with known characteristics 

of BMSCs. In the BMSC group 4 weeks after BMSC infusion, 

the area of Sirius red-stained liver fibrosis was significantly 

reduced (fibrotic area (%); BMSCs: −1.62 ± 0.63; controls: 

+1.44 ± 1.01, p < 0.05), consistent with a significantly shortened 

half-life of ICG (half-life of ICG (min); BMSCs: −1.45 ± 

0.42; controls: +1.23 ± 0.87, p < 0.05). After BMSC infusion, 

no pro-coagulation activity and no thrombosis were seen in 

lung arteries using contrast-enhanced computed tomography. 

Conclusions: We established a useful canine liver cirrhosis 

model with repeated CCl 4 

administration through a PICC and 

confirmed that cultured autologous BMSC infusion improved 

liver cirrhosis and promoted liver regeneration without adverse 

effects. 

Disclosures: 

The following authors have nothing to disclose: Takashi Matsuda, Taro Takami, 

Yuki Aibe, Toshihiko Matsumoto, Tsuyoshi Ishikawa, Naoki Yamamoto, Shuji 

Terai, Isao Sakaida 

1448 

A novel glycobiomarker, Wisteria floribunda agglutinin 

Macrophage Colony-Stimulating Factor Receptor can 

predict carcinogenesis and survival of liver cirrhosis 

with hepatitis C virus 

Etsuko Iio 1,3 , Makoto Ocho 2 , Akira Togayachi 2 , Masanori 

Nojima 4 , Atsushi Kuno 2 , Masashi Mizokami 5 , Hiroshi Yatsuhashi 6 , 

Tatsuya Ide 7 , Shunsuke Nojiri 3 , Hisashi Narimatsu 2 , Yasuhito 

Tanaka 1 ; 1 Virology and Liver Unit, Nagoya City University Graduate 

School of Medical Sciences, Nagoya, Japan; 2 Research Center 

for Medical Glycoscience, National Institute of Advanced Industrial 

Science and Technology, Tsukuba, Japan; 3 Gastroenterology and 

Metabolism, Nagoya City University Graduate School of Medical 

Sciences, Nagoya, Japan; 4 Advanced Medicine Promotion, 

The Advanced Clinical Research Center, The Institute of Medical 

Science, University of Tokyo, Tokyo, Japan; 5 The Research Center 

of Japan, Hepatitis and Immunology, Kohnodai Hospital, International 

MedicalCenter, Ichikawa, Japan; 6 Clinical Research Center, 

National Nagasaki Medical Center, Omura, Japan; 7 Division of 

Gastroenterology, Department of Medicine, Kurume University, 

Kurume, Japan 

[Background/Aims] Recently, we identified a novel liver fibrosis 

glycobiomarker Wisteria floribunda agglutinin (WFA)-reactive 

colony stimulating factor 1 receptor (WFA + -CSF1R) using a glycoproteomics-based 

strategy. The aim of the present study was 

to assess the value of measuring WFA + -CSF1R levels for hepatocarcinogenesis 

and outcome in liver cirrhosis (LC) patients 

with hepatitis C virus (HCV). [Methods] WFA + -CSF1R and total 

CSF1R levels were measured by an antibody-lectin sandwich 

ELISA in serum samples from 214 consecutive HCV infected 

patients (99 CH and 115 LC) from January 1998 to April 2013 

in order to evaluate impact on carcinogenesis and survival of 

LC patients. Among 115 patients with LC, 56 (48.7%) patients 

without hepatocellular carcinoma (HCC). Ultimately, the 56 LC 

patients and independent 45 LC patients without HCC were 

assessed. [Results] LC patients without HCC of training cohort 

(n = 56) and validation cohort (n = 45) at baseline were analyzed 

for survival and carcinogenesis rates. The optimal cut-off 

of the WFA + -CSF1R were determined to be 310 ng/ml according 

to the smallest P value screened by Cox regression analysis 

in the training set. The AUC of WFA + -CSF1R for predicting 

overall survival, calculated by time-dependent ROC analysis, 

was 0.691 and the HR (per 1-SD increase) was 1.80 (95% CI, 

1.23-2.62, p < 0.001) of combined with the training set and 

validation set. The survival rate of LC patients with high WFA + - 

CSF1R levels (≥ 310 ng/ml) was significantly worse than those 

with lower levels (p < 0.01). Next, we analyzed the percentage 

of WFA + -CSF1R among total-CSF1R (WFA + -CSF1R%) for 

the potential of HCC. The optimal cut-off for the WFA + -CSF1R% 

for predicting the cumulative carcinogenesis rate was 35.0%. 

We further verified the utility of WFA + -CSF1R%, using the validation 

set. In the combined data the AUC of WFA + -CSF1R% for 

predicting the cumulative carcinogenesis rate was 0.760, with 

an HR of 1.66 (95% CI 1.26-2.20, p < 0.001). The AUC of 

WFA + -CSF1R% was superior to other fibrosis and tumor markers 

(i.e. Fib4, APRI, albumin, AFP, AFP-L3 and PIVKA-II) for predicting 

the cumulative carcinogenesis rate. LC patients in the 

training cohort with high WFA + -CSF1R% (≥ 35.0%, n = 6) had 

cumulative carcinogenesis rates of 33.3%, 50.0%, and 75.0% 

at 1, 3 and 5 years, respectively. In contrast, patients with 

low WFA + -CSF1R% (< 35.0%, n = 50) had cumulative rates 

of 4.6%, 18.0%, and 35.0%, respectively (p = 0.006). These 

data were supported in the validation cohort. [Conclusions] 

Assessing serum levels of WFA + -CSF1R has diagnostic value 

for predicting carcinogenesis and the survival of LC patients.


Disclosures: 



Squibb 

The following authors have nothing to disclose: Etsuko Iio, Makoto Ocho, Akira 

Togayachi, Masanori Nojima, Atsushi Kuno, Masashi Mizokami, Hiroshi Yatsuhashi, 

Tatsuya Ide, Shunsuke Nojiri, Hisashi Narimatsu 

1449 

Second harmonic generation microscopy based quantitative 

assessment for HBV-associated liver fibrosis 

Kai-Wen Huang; National Taiwan University Hospital, Taipei, Taiwan 

Background Liver fibrosis is associated with an abnormal 

increase in an extracellular matrix in chronic liver diseases. 

Quantitative characterization of collagen in intact tissue is 

essential for both fibrosis studies and clinical applications. 

Commonly used methods, histological staining followed by 

either semi-quantitative or computerized image analysis, have 

limited sensitivity, accuracy, and operator-dependent variations. 

The collagen in livers could be observed through second-harmonic 

generation (SHG) microscopy. The two photon 

excited fluorescence (TPEF) images, recorded simultaneously 

with SHG, clearly revealed the hepatocyte morphology and 

associated collagen progression dynamics. Purpose To access 

liver fibrosis using SHG/TPEF microscopy on human liver 

biopsy and resection samples, and to identify and quantitatively 

measure the morphological features, and to generate 

as an index for describing the severity of fibrosis. Methods A 

total of 95 specimens were sampled from patients with hepatitis 

B viral infections underwent liver biopsy or liver resection 

in National Taiwan University Hospital between 2010-2012. 

Liver fibrosis was graded by pathologist in NTUH using the 

Metavir scoring system. Quantitative analysis was also done 

on the same sample using SHG/TPEF microscopy to generate 

100 quantitative measurements on morphological features. 

These 100 quantitative measurements are used to develop the 

algorithm to fit the staging results from Metavir using non-linear 

support vector machine (SVM) model. Results Out of 100 quantitative 

measurements, 53 showed strong correlation with Metavir 

staging and could statistically distinguish patients of stage 

0 from stage 1, and 67 for distinguishing patients of stage 3 

from stage 4. Out of these quantitative measurements, 10 were 

selected to develop the algorithm using SVM model using half 

of the data for model training, and area under curve (AUC) 

generated from the remaining half samples showed 0.76 with 

these 10 selected features. Conclusion Quantitative assessment 

methodology combining non-stain SHG/TPEF imaging technology 

and non-linear fitting model showed great correlation 

with traditional pathology staging system. This process can be 

automated to minimize the potential inter and intra observer 

discrepancies and give highly quantitative and reproducible 

results for clinical diagnosis and treatment response assessment. 

Disclosures: 

The following authors have nothing to disclose: Kai-Wen Huang 

1450 

Hepatitis C treatment and Progression of Liver Stiffness 

in Australia, subsequent to the introduction of protease 

inhibitors 

Yang Wu 1 , Hank H. Chen 1 , Martin Weltman 1 , Guy D. Eslick 2 , 

Bilel Jideh 1 , Izabella Pokorski 1 ; 1 Gastroenterology, Nepean Hospital, 

Kingswood, NSW, Australia; 2 Surgery, Nepean Hospital, 

Kingswood, NSW, Australia 

Background: Transient elastography (TE) is a noninvasive and 

well-validated method for measurement of liver stiffness in 

patients with chronic Hepatitis C. Aim: In the previous era of 

interferon-based therapy, studies have shown a strong association 

between Sustained Virological Response (SVR) and 

improvement in Liver stiffness (LS) 1 .Our current study aims 

to explore the kinetics of liver stiffness in Australian patients 

receiving hepatitis C treatment including protease inhibitors, an 

area not examined previously. Method: Consecutive patients at 

Nepean Hospital treated for Hepatitis C from 2011 onwards 

were included in the study. Patients were treated according to 

standard of care for their respective genotypes, which included 

protease inhibitors in patients with Genotype 1. The patient’s 

initial LS measurement was taken prior to their treatment and 

a second measurement was made at an interval at least 12 

weeks after the end of their treatment. Other patient factors 

such as gender, age, presence or absence of cirrhosis, alcoholism 

and blood tests were also collated. Results: Of the 36 

patients that were included in the study, 26 (72%) patients 

achieved SVR. Overall, 21(58%) patients were treated with 

protease inhibitors. The mean intra-patient change relative to 

baseline at the follow-up elastography was -4.97 kPa in the 

patient group who achieved SVR, vs +1.55 kPa in the patients 

who did not achieve an SVR (p = 0.07056). In uni-variate 

analysis (Mann-Whitney U test), other parameters such as protease 

inhibitor use (p=0.29), viral genotype, treatment experience 

(p=0.89), gender (p=0.09), age, alcohol intake (p=0.62) 

and presence of cirrhosis (p=0.94) were not predictive of LS 

improvement. Similarly, viral load and ALT measurement at 

the start of treatment didn’t reveal any significant relationship 

with post-treatment LS during Logistic regression. Conclusion: 

Measurement of liver stiffness with TE after hepatitis C treatment 

showed that although the SVR group had more improvement in 

the liver stiffness score compared to the non-SVR group, the difference 

was not statistically significant in this study. Our study 

is one of the first to include patients treated with protease inhibitors. 

References: Andersen ES, Moessner BK, Christensen PB, 

Kjær M, Krarup H, Lillevang S, Weis N. Lower liver stiffness 

in patients with sustained virological response 4 years after 

treatment for chronic hepatitis C. Eur J Gastroenterol Hepatol. 

2011 Jan;23(1):41-4. 

Disclosures: 

The following authors have nothing to disclose: Yang Wu, Hank H. Chen, Martin 

Weltman, Guy D. Eslick, Bilel Jideh, Izabella Pokorski


1451 

The Attention-to-Burden Index (ABI): a New Tool that 

Reveals Profound Disparities between Research Attention 

and Disease Burden among Liver Diseases. 

Nambi J. Ndugga 1 , Teisha G. Lightbourne 2 , Kavon Javaherian 2 , 

Neha Verma 2 , Eric S. Orman 3 , David Pesci 2 , Ramon Bataller 4,2 ; 

1 Center for Gastrointestinal Biology and Disease, University of 

North Carolina at Chapel Hill School of Medicine, Chapel Hill, 

NC; 2 Nutrition, University of North Carolina at Chapel Hill, Chapel 

Hill, NC; 3 University of North Carolina at Chapel Hill, Chapel Hill, 

NC; 4 Medicine, University of North Carolina at Chapel Hill School 

of Medicine, Chapel Hill, NC 

Rationale: In recent years, there have been great advances 

in the management of HBV and HCV. Conversely, there are 

no approved therapies for fatty liver diseases (NAFLD and 

ALD). We hypothesize that these unequal advances are due 

to disparities in the research attention surrounding different 

liver diseases. Methods: To test this hypothesis, we developed 

the Attention-to-Burden Index (ABI), a comprehensive tool comprised 

of main research activities, and an estimate of disease 

burden for the main types of liver diseases: HCV, HBV, ALD, 

NAFLD. A systematic study of the research attention was carried 

out for the years 2010-2015 on these 4 liver diseases. The 

mean research attention for each liver disease was calculated 

from 5 different levels: reference in the two major annual liver 

meetings (AASLD and EASL), number of drugs in the pipeline of 

38 major pharmaceutical companies, funded grants (USA and 

EU), ongoing clinical trials (ClinicalTrials.gov) and scientific 

publications (PubMed). The mean disease burden was calculated 

from the following parameters: hospitalization costs in 

USA, etiology of liver transplantation in USA and Europe, etiology 

of cirrhosis, and etiology of liver fibrosis. The ABI for each 

liver disease was calculated as the mean attention divided by 

the mean disease burden. Results: The research attention in 

both AASLD and EASL meetings to the main liver diseases was 

similar: HCV (55 and 54% of total attention, respectively), HBV 

(26 and 26%), NAFLD (13 and 15%), and ALD (6 and 4%). 

The number of drugs in the pipeline with specific indications 

for liver diseases was: 13 for HCV, 4 for HBV, 3 for NAFLD, 

1 for ALD. The percentage of funded grants in the US and EU 

was: HCV (54 and 62% of total grants, respectively), HBV 

(24 and 33%), NAFLD (17 and 5%), and ALD (5 and 1%). 

A similar pattern of research attention was found in ongoing 

clinical trials and scientific publications. The calculated overall 

burden for the main liver disease was 34% for HCV, 4% for 

HBV, 37% for ALD and 24% for NAFLD. When comparing the 

ratio between the overall research attention and the disease 

burden of main liver diseases, we found striking differences. 

The calculated ABI for HCV and HBV revealed a 1 and 7-fold 

over-attention, respectively, while NAFLD and ALD received an 

infra-attention of 2 and 8-fold. Conclusions: The ABI reveals disparities 

between research attention and disease burden among 

liver diseases, with a clear over-attention to viral hepatitis compared 

to fatty liver diseases. Notably, the attention to ALD is 

minimal. There is a critical need to increase awareness of ALD 

in the liver research community. 

Disclosures: 

Ramon Bataller - Advisory Committees or Review Panels: Sandhill; Consulting: 

VTI, Oncozyme Pharma 

The following authors have nothing to disclose: Nambi J. Ndugga, Teisha G. 

Lightbourne, Kavon Javaherian, Neha Verma, Eric S. Orman, David Pesci 

1452 

Hospital Based Initiative to Decrease Readmission Rates 

in End-Stage Liver Disease 

Sheela S. Reddy 1 , Chad Gorn 2 , Stephania Dottin 2 , Ann E. Burke 1 , 

Karen Newell 2 , Karen Pine 2 , Ursula Hobbs 2 , Maria Neff 2 , Jonathan 

M. Fenkel 1 , David A. Sass 1 , Steven K. Herrine 1 , Jesse M. 

Civan 1 , She-Yan Wong 1 , Dina Halegoua-De Marzio 1 ; 1 Medicine, 

Division of Gastroenterology and Hepatology, Thomas Jefferson 

University, Philadelphia, PA; 2 Transplant Services, Thomas Jefferson 

University Hospital, Philadelphia, PA 

Introduction: The Hospital Readmission Reduction Program, a 

part of the Affordable Care Act, requires reduced payments to 

hospitals with excess readmissions, or admission to a hospital 

within 30 days of discharge. This penalization is based on a 

hospital’s expected readmission rate, an adjusted value of the 

national average. Studies have shown a ~37% readmission 

rate for patient with end stage liver disease (ESLD). Due to the 

burden of ESLD readmissions at our institution, a care coordination 

(CC) program was created to improve transitions of 

care from the inpatient to outpatient setting. The aim of this 

study is to assess whether the implementation of the CC program 

reduced 30-day readmissions. Methods: Our institution 

is an urban quaternary care liver transplant center performing 

approximately 50 liver transplants yearly in highly competitive 

UNOS Region 2. Our CC program was created February 

2014, with a focus on patient education, short-term outpatient 

follow-up visits, and post-discharge follow-up calls by a transplant 

nurse coordinator. During inpatient admission, patients 

deemed high-risk by attending hepatologists were enrolled. 

High-risk was defined as cirrhotic patients admitted with fluid 

overload, hepatic encephalopathy and/or renal disease requiring 

hemodialysis. Once discharged, the patient was scheduled 

for an office visit within 7 days and received follow-up phone 

calls at 48 hours after discharge, and on days 7, 14, 21, and 

30. Patients who were transplanted, transferred, or expired 

before 30 days were excluded from final analysis. Results: Our 

hepatology service readmission rate was 33.67% in 2013. 

Since February 2014, 90 high-risk patients were identified and 

enrolled in the CC program. Of these 90 high-risk patients, 

30-day readmission was prevented in 37.9% of patients, but 

62.1% still had at least one 30-day readmission. CC protocol 

compliance was 87.5%. From October 2014 to March 2015, 

encephalopathy was the most likely reason for readmission 

(13 admissions), followed by bleeding (6), infection (6), and 

shortness of breath (6). There was no improvement seen in the 

hepatology readmission rate, with an overall rate of 36% in 

Q4 of 2014. Conclusion: Preventing readmission in patients 

with ESLD and high-risk decompensating events remains challenging, 

especially in a high MELD at transplant region. Liver 

transplantation is the only way to reduce readmissions in this 

population, as the natural history of ESLD lends itself to frequent 

hospitalization in the last stages of the disease. The target of a 

hospital-based initiative to prevent readmission should perhaps 

target those with a moderate risk for readmission in order to 

have an impact. 

Disclosures: 

Jonathan M. Fenkel - Advisory Committees or Review Panels: Merck; Consulting: 

Gilead Pharmaceuticals, Janssen Therapeutics, Bristol-Myers Squibb, Abbvie 

Steven K. Herrine - Board Membership: ABIM; Grant/Research Support: BMS, 

Gilead, Merck, Vertex 


The following authors have nothing to disclose: Sheela S. Reddy, Chad Gorn, 

Stephania Dottin, Ann E. Burke, Karen Newell, Karen Pine, Ursula Hobbs, Maria 

Neff, David A. Sass, She-Yan Wong, Dina Halegoua-De Marzio


1453 

Cost-effectiveness of Sofosbuvir plus Ledipasvir Antiviral 

Treatment for Elderly Patients with Chronic Hepatitis C 

Genotype 1 

Antonio Ciaccio 2 , Paolo Cortesi 3 , Giuseppe Bellelli 4 , Matteo 

Rota 5 , monica Rota 2 , Lorenzo G. Mantovani 3 , Giorgio Annoni 4 , 

Mario Strazzabosco 1,2 ; 1 Yale University, Section of Digestive Diseases, 

New Haven, CT; 2 Department of Surgery and Translational 

Medicine, University of Milano-Bicocca, Milan, Italy; 3 Research 

Centre on Public Health (CESP), University of Milan-Bicocca, 

Milan, Italy; 4 Department of Health Science, University of Milan-Bicocca, 

Milan, Italy; 5 Department of Epidemiology, IRCCS-Istituto di 

Ricerche Farmacologiche Mario Negri, Milan, Italy 

A relevant proportion of patients affected by Chronic Hepatitis 

C (CHC) is older than 65 years. In the interferon era, comorbidities 

and a higher susceptibility to interferon/ribavirin adverse 

events have limited treatment in these patients. Recent approval 

of interferon-free regimens, with high efficacy and limited toxicity, 

provide unprecedented chances for these patients to be 

treated. However, the cost-effectiveness of all-oral Direct-Acting 

Antivirals (DAAs) has not been addressed in the elderly 

population. We have performed a cost-effectiveness analysis 

taking into account the severity of liver disease, the age of the 

patient and the geriatric (frailty) status. METHODS: A semi-Markov 

model of CHC natural history was built. The study focuses 

on CHC Genotype 1 patients older than 65 years, stratified 

according to liver fibrosis (METAVIR F3 and F4), age (65 to 90 

years old) and frailty phenotype defined by Fried’s (not frail, 

pre-frail and frail) for a total of 30 cohorts simulated. Treatment 

with sofosbuvir plus ledipasvir (SOF/LDV) versus no treatment 

was assessed for each cohort. The model estimated costs, Life 

Years and Quality-Adjusted Life Years (QALY) using a lifetime 

time horizon and the Health System perspective. Results are 

presented as incremental cost-effectiveness ratios (ICERs) per 

QALY gained. Cost-effectiveness was defined as an ICER under 

the willingness-to-pay threshold of 40,000 € per QALY gained. 

RESULTS: At each fibrosis score, ICER increased with age and 

frailty index. Among patients with F3 fibrosis, ICER ranged from 

€5,530/QALY in not-frail 65 years old and €149,355 in frail 

85 years old patients. Among F4 patients ICER ranged from 

€6,552/QALY in not frail 65 years old and €85,559 in frail 

85 years old patients. In F3 and F4 cohorts ICER was below 

€40,000/QALY up to age 84 and 87 years in non-frail pts, up 

to age 80 and 83 years in pre-frail patients, and up to age 76 

and 80 in frail patients, respectively. Adopting an alternative 

scenario with a 20% discount of SOF/LDV treatment, cost-effectiveness 

increases accordingly. CONCLUSION: This analysis 

shows that, assuming that efficacy and safety of SOF/LDV are 

not age-dependent, SOF/LDV treatment is cost-effective in most 

CHC patients older than 65 years, although a careful assessment 

of the patient status is mandatory. Furthermore, under 

conditions of restricted budget, it should be considered that 

ICER varies considerably between two non-frail F4 patients, 

one at age 65 and the other at 85 years (between 8,000 and 

33,000 EUR/QALY, respectively). This cost-effectiveness analysis 

should promote a prospective study to verify efficacy and 

side effects in elderly HCV patients. 

Disclosures: 

Antonio Ciaccio - Grant/Research Support: Gilead Sciences 

Lorenzo G. Mantovani - Advisory Committees or Review Panels: Bayer; Grant/ 

Research Support: Jansen, Merck and Co; Speaking and Teaching: Bayer 

The following authors have nothing to disclose: Paolo Cortesi, Giuseppe Bellelli, 

Matteo Rota, monica Rota, Giorgio Annoni, Mario Strazzabosco 

1454 

Living Donor Liver Transplants (LDLT), compared to 

Deceased Donor Liver Transplants (DDLT), and MELD 

Score Increase Liver Transplant Resource Utilization at 

an Experienced Transplant Center from June 2008 to 

July 2013 

Grace Kim 1 , Yael J. Coppleson 2 , Robert S. Brown 3,2 , Thresiamma 

Lukose 1 , Jean C. Emond 1,2 , Benjamin Samstein 2 ; 1 Surgery, Columbia 

University, New York, NY; 2 NewYork Presbyterian, New York, 

NY; 3 Medicine, Columbia University, New York, NY 

Purpose: To examine liver transplant charges at an experienced 

transplant center. Methods: This retrospective analysis 

used data for liver transplant recipients at NewYork-Presbyterian 

from hospital billing charges, medical records, and 

UNOS. The patient population (n=526) included both adult 

(n=458) and pediatric patients (n=68) undergoing a first liver 

transplant, but excluded combined transplants and status 1 

listed patients. Total hospital charges were used as a proxy 

for resource utilization, and included all clinical services and 

resources for transplant day, and inpatient and outpatient preand 

post-transplant care. Transplant day charges included both 

donor and recipient operating and recovery services. Multivariate 

linear regressions were utilized to examine the impact of 

living donor transplantation and MELD on transplant day and 

total transplant charges, controlling for gender and age, and 

diagnosis of HCC. Results: The average overall MELD score 

of the cohort was 24.3, while the average MELD was 25.4 

for DDLT (n=474), and 18.1 for LDLT (n=87) (p=0.00). Age, 

gender, and etiology of liver disease were similar across donor 

type. For adult patients, transplant day charges were greater 

for LDLT patients (p=.04) and higher MELD patients (p=0.00). 

Total charges were greater but not statistically significant 

(p=.14) for LDLT patients, though still increased by each MELD 

point (p=0.00). For all patients, transplant day charges were 

higher for LDLT patients (p=.04) and higher MELD patients, 

($13K per point, p =0.00). For total costs, LDLT charges were 

higher (p=.16) and MELD point increased charges (p=.01). 

The difference between LDLT and DDLT in total transplant 

charges were equalized by an 8-point differential in MELD 

score. Conclusion: Living donor transplants are associated with 

greater transplant resource utilization, but the difference in total 

transplant charges is not statistically significant. The higher 

charges associated with the increased complexity of LDLT cases 

are mitigated by shorter waiting time and fewer post-operative 

days until discharge. Patients listed with higher MELD scores 

utilize significantly more resources for transplant care. 

Disclosures: 


The following authors have nothing to disclose: Grace Kim, Yael J. Coppleson, 

Thresiamma Lukose, Jean C. Emond, Benjamin Samstein


1455 

A probabilistic decision model using non-invasive fibrosis 

markers in Primary Care NAFLD pathways predicts 

increased cirrhosis detection rates and reduced overall 

healthcare expenditure 

Ankur Srivastava 1,2 , Simcha Jong 3 , Anna Gola 4 , Laura Fenlon 5 , 

Petra Scantlebury 5 , Sudeep Tanwar 1,2 , Hannah Liu 7 , Ruth E. 

Gailer 6,7 , Sarah Morgan 6 , Alex Warner 6 , Karen Sennett 7 , Julie 

Parkes 1 , James O’Beirne 1,2 , Emmanuel Tsochatzis 1,2 , William M. 

Rosenberg 1,2 ; 1 Institute of Liver and Digestive Health, University 

College London (UCL), London, United Kingdom; 2 Hepatology, 

Royal Free London NHS Foundation Trust, London, United Kingdom; 

3 Management Science & Innovation, University College 

London, London, United Kingdom; 4 Health Economics, University 

College London, London, United Kingdom; 5 Department of 

Finance, Royal Free London NHS Foundation Trust, London, United 

Kingdom; 6 Primary Care, Camden Clinical Commissioning Group, 

London, United Kingdom; 7 Primary Care, Islington Clinical Commissioning 

Group, London, United Kingdom 

Background: Risk factors for Non-Alcoholic Fatty Liver Disease 

(NAFLD) have reached epidemic proportions and whilst only 

a minority of at-risk individuals develop significant liver disease 

warranting specialist referral, ensuring identification of 

this group is a primary care challenge. Non-invasive tests (NIT) 

for liver fibrosis enable community-based doctors to identify 

patients with NAFLD who have advanced fibrosis and would 

benefit from early referral for specialist care and interventions 

to limit the complications of cirrhosis. As part of an evaluation 

of a novel primary care NAFLD pathway in two London boroughs 

(Camden and Islington, C&I), which uses NIT to stratify 

patients for advanced liver fibrosis, we have constructed an 

analytical model to examine the potential cost implications of 

this strategy on an intention-to-treat basis. Methods: A probabilistic 

decision analytical model was constructed to explore 

the financial dimensions of two strategies in community based 

management of NAFLD from a healthcare payer perspective: 

1) Standard of care (SOC) comprising physicians’ clinical judgment 

based on clinical history, examination, blood tests and 

ultrasound 2) SOC plus Non-Invasive Testing for liver fibrosis 

using FIB-4, and the ELF test (to resolve indeterminate FIB-4 

tests) to guide referral. The model was populated from the literature, 

national UK data and expert opinion. A five year time 

horizon was applied. Results: The C&I population is 434,958. 

It was assumed that 30% of the NAFLD population consult their 

general practitioner (GP) annually. >F2 fibrosis prevalence in 

NAFLD was set at 5%. Local clinical audit estimated SOC sensitivities 

and specificities of 0.35 and 0.70. The model estimated 

24% needed ELF test. Utilizing NIT reduced referrals of 

≤F2 disease by over 80% and resulted in over 50% increase 

in the detection of cirrhosis. There was a significant reduction 

in liver transplantation rates by approximately 15%. The 

overall expenditure was reduced by a fifth, primarily through 

reductions in referrals for low risk cases, costs related to HCC 

management and emergency inpatient admissions. Discussion: 

The use of NIT led to a reduction in inappropriate referrals and 

improvement in the early detection of cirrhosis and its complications. 

These changes are likely to result in significant health 

gains and economic cost savings. With NAFLD prevalence 

likely to increase, policy needs to address these challenges. 

Modeling suggests that the Camden and Islington NAFLD pathway 

employing FIB-4 and ELF to guide GPs’ risk stratification 

of patients will achieve these goals. A full clinical effectiveness 

and cost consequence analysis is underway. 

Disclosures: 

Simcha Jong - Grant/Research Support: Siemens 





The following authors have nothing to disclose: Ankur Srivastava, Anna Gola, 

Laura Fenlon, Petra Scantlebury, Sudeep Tanwar, Hannah Liu, Ruth E. Gailer, 

Sarah Morgan, Alex Warner, Karen Sennett, James O’Beirne, Emmanuel Tsochatzis 

1456 

Impact of Daclatasvir-Sofosbuvir Combination Treatment 

on Medical Events and Costs in Patients Infected with 

Genotype 3 Hepatitis C Virus 

Boris Gorsh 1 , Bruce E. Sill 1 , Shalini Hede 1 , Catherine St-Laurent 

Thibault 2 , Divya Moorjaney 3 , Michael Ganz 3 , Anupama 

Kalsekar 1 , Yong Yuan 1 ; 1 Bristol-Myers Squibb, Plainsboro, NJ; 

2 Evidera, Montreal, QC, Canada; 3 Evidera, Boston, MA 

Purpose: Patients infected with genotype (GT) 3 of the hepatitis 

C virus (HCV) fail antiviral therapy more often than patients 

infected with other GTs. Combination daclatasvir-sofosbuvir 

(DCV+SOF) is a significant advancement as it requires 12 

rather than 24 weeks for other combination therapies. Our 

objective is to estimate the number of medical events and 

direct medical costs avoided if, otherwise untreated, GT3 HCV 

patients were treated with DCV+SOF. Methods: We used a 

published Markov model [i] to estimate the number of medical 

events and costs experienced by a hypothetical cohort of 

1,000 HCV GT3 patients (mean age of 54 years, 65% male) 

who received DCV+SOF or no treatment. The patient distribution 

we used was based on McGarry et al (2012). [ii] We 

assumed that 89% of the DCV+SOF-treated patients would 

achieve a sustained viralogic response based on results of the 

ALLY-3 study. Medical events and costs were assessed for three 

scenarios: overall HCV population (F0-F4) receives no treatment, 

only the F3/F4 subpopulation receives DCV+SOF, and 

all patients (F0-F4) receive DCV+SOF. Total medical events 

were defined as the sum of compensated cirrhosis, decompensated 

cirrhosis, hepatocellular carcinoma, liver transplant, or 

liver-related death. Medical costs include monitoring costs and 

costs for treating complications and adverse events. Disease 

progression and costs were modelled over time horizons of 20 

and 50 years. Results: Medical events and direct medical costs 

were lowest when all patients were treated with DCV+SOF (76 

events; $14,343,786). When only F3/F4 patients received 

DCV+SOF, medical events and costs were substantially higher 

(335 events; $68,414,213). In the scenario where no one 

was treated, events and costs were the highest (729 events, 

$129,838,658). Similar results were seen over 50 years. 

Conclusion: Substantially fewer medical events and lower 

medical costs are accrued when all GT3 patients are treated 

with DCV+SOF compared with scenarios in which no one is 

treated or treatment is limited to F3/F4 patients. [i] McEwan P, 

Ward T, Yuan Y, et al. The impact of timing and prioritization 

on the cost-effectiveness of birth cohort testing and treatment 

for hepatitis C virus in the United States. Hepatology. 2013 

Jul;58(1):54-64. [ii] McGarry LJ, et al. Economic model of a 

birth cohort screening program for hepatitis C virus. Hepatology. 

2012 May;55(5):1344-55. 

Medical Events and Costs (Per Cohort) 

Disclosures: 

Boris Gorsh - Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers 

Squibb


Bruce E. Sill - Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers 

Squibb 

Anupama Kalsekar - Employment: Bristol Myers Squibb 

Yong Yuan - Employment: Bristol Myers Squibb Company 

The following authors have nothing to disclose: Shalini Hede, Catherine St-Laurent 

Thibault, Divya Moorjaney, Michael Ganz 

1457 

Hospitalisation for primary biliary cirrhosis: a UK-PBC 

analysis of hospital episodes 

Gideon Hirschfield 2 , Luke Vale 3 , Tracy J. Mayne 1 , George F. 

Mells 4 , David Shapiro 1 , David Jones 5 ; 1 Intercept Pharmaceuticals, 

San Diego, CA; 2 Centre for Liver Research, University of Birmingham, 

Birmingham, United Kingdom; 3 Institute of Health & Society, 

Newcastle University, Newcastle, United Kingdom; 4 Division of 

Gastroenterology and Hepatology, Department of Medicine, University 

of Cambridge, Cambridge, United Kingdom; 5 Institute of 

Cellular Medicine, Newcastle University Medical School, Newcastle, 


Background: The advent of new therapies for patients with 

primary biliary cirrhosis (PBC) highlights the need to understand 

the current health burden of PBC. Objective: To assess 

the frequency and nature of hospitalisations associated with 

primary biliary cirrhosis (PBC). Methods: UK-PBC analysed all 

records from 2009 through 2014 in the Hospital Episodes Statistics 

database, containing information on all hospitalisation 

across the National Health Service in England, where the ICD- 

10 code for PBC (K74.3) appeared. We characterised primary 

diagnosis for each hospitalisation. Results: There were 21,275 

admissions with a PBC code over the 5-year observational 

period covering 1631 unique ICD-10s listed as the primary 

hospitalisation code. PBC was the primary code for 17% of 

admissions. The number of relevant hospitalisations increased 

from 3368 in 2009/2010 to 4995 in 2012/2014. The number 

per 100,000 hospitalisations increased from 23 to 32 over 

this period (Figure). The increase was almost completely driven 

by the inclusion of PBC as a secondary diagnosis. Liver transplants 

represented 5-10% of admissions with PBC as a primary 

code, and 1- 2% of any PBC-related admission. The next 20 

ICD-10 codes accounted for an additional 17% of admissions. 

The top 20 codes included known sequelae of PBC: ascites 

(2%), varices (2%), and liver cancer (1%). Remaining codes 

included: anaemia (iron deficiency and unspecified) 3%; respiratory 

(pneumonia, COPD and lower respiratory infections) 

3%; osteoporosis and fractures 2%; and various abdominal 

(unspecified abdominal pain, gastritis and gastroenteritis) 2%. 

Conclusion: Our UK-PBC data confirms a significant clinical 

need for patients with PBC, including a rising healthcare burden 

from disease. The increase in PBC-related diagnoses may 

be due to more accurate coding, greater co-morbidity as the 

result of increased longevity, or may represent the true impact 

of disease. New therapies preventing the progression of PBC to 

end-stage cirrhosis, and its complications, may help to reduce 

this rising burden. 

Disclosures: 





Falk, Shire 

The following authors have nothing to disclose: Gideon Hirschfield, Luke Vale, 

George F. Mells 

1458 

Obeticholic Acid (OCA) has Incremental Efficacy in 

Patients with Higher Baseline Alkaline Phosphatase 

(ALP) Levels 

Tracy J. Mayne, Richard Pencek, Tonya Marmon, David Shapiro; 

Intercept Pharmaceuticals, Inc., San Diego, CA 

Objective: Examine ALP lowering stratified by baseline ALP 

among patients with primary biliary cirrhosis (PBC) treated with 

10 mg OCA. Methods: We pooled data from the 10 mg treatment 

arms of two phase 2 trials (n=20 and n=38) and one 

phase 3 trial (n=73) randomized, double-blind, placebo-controlled 

trials of OCA in patients with PBC. The percent change 

in ALP at 3 months was calculated based on baseline ALP stratified 

by 50 U/L. We ran a linear regression predicting percent 

change in ALP based on baseline ALP. Results: As shown in the 

first figure, both absolute and percent reduction in ALP were 

greater at higher baseline ALP levels. As shown in the second 

figure, for each incremental 100 U/L in baseline ALP, there 

was an incremental 5% reduction in ALP at 3 months (intercept 

= 17%) in patients treated with 10 mg OCA. Conclusion: A 

standard measure of drug efficacy is percent reduction in a 

specified marker. Statin efficacy is measured as percent reduction 

in low density lipid cholesterol (LDL-C); blood pressure 

medications are assessed by percent reduction in systolic and 

diastolic blood pressure. At a fixed percent reduction, absolute 

reduction in a biomarker will be greater with higher baseline 

levels: a 50% reduction in LDL-C at a baseline of 200 mg/dL 

is 100 mg/dL; a 50% reduction at 100 mg/dL is 50 mg/dL. 

In the case of OCA, the percent reduction in ALP increases as 

a function of baseline ALP. As shown by the regression model, 

OCA produces an average 59% reduction at a baseline ALP 

of 800 U/L, and an average 38% reduction at a baseline ALP 

of 400 U/L. Achieved level of ALP has been shown to be associated 

with improved progression-free survival (Lammers et al, 

Gastroenterology 2014;147:1338). These data indicate that 

patients with more progressed PBC, as measured by higher 

serum ALP, may receive incremental benefit when treated with 

OCA.


Disclosures: 








1459 

Impact of Comorbidity on Inpatient Outcomes for 

Decompensated Cirrhosis 

Brandon Nokes 2,1 , Amit Kashyap 3 , Thomas D. Boyer 4,3 , Archita P. 

Desai 4,3 ; 1 Internal Medicine, Mayo Clinic Arizona, Phoenix, AZ; 

2 College of Medicine, University of Arizona, Tucson, AZ; 3 Gastroenterology 

and Hepatology, University of Arizona, Tucson, AZ; 

4 Liver Research Institute, University of Arizona, Tucson, AZ 

Background: Decompensated cirrhosis is an increasingly 

common reason for admission. Inpatients with cirrhosis and 

comorbidities have worse outcomes. While current comorbidity 

indices used for risk adjustment report number of comorbidities, 

the complex interaction between cirrhosis, comorbidity and outcomes 

may be better understood by capturing naturally occurring 

subgroups of cirrhosis patients with common comorbidity 

profiles. Aim: We sought to describe inpatient health care utilization 

due to decompensated cirrhosis and to create comorbidity 

profiles for patients admitted with decompensated cirrhosis. 

Methods: We examined the 2008-2011 NIS and included all 

admissions with a diagnosis of cirrhosis and a complication of 

cirrhosis. Means and proportions with standard errors and confidence 

intervals were used to describe the sample. Latent class 

mixture modeling was used to identify distinct and common 

comorbidity profiles among binary categorical variables indicating 

the presence or absence of comorbidities. Generalized 

linear model analysis was used to quantify associations among 

comorbidity profiles and outcomes. Results: Of the full NIS, 

414521 (1.3%) admissions had a cirrhosis diagnosis of which 

237553 (57%) met inclusion criteria. The average patient was 

58 years old, white (58%), male (62%) with hepatitis C (29%) 

or alcohol related liver disease (51%), insured by Medicare 

(40%) and cared for in a large (66%), non-teaching (51%), 

rural (90%) hospital. Complications of cirrhosis were most often 

ascites (57%), portal hypertension (40%), hepatic encephalopathy 

(28%), and acute renal failure (23%). Compared to 

those with Elixhauser comorbidity index of 0, those with an 

index of 3 or more had 72% higher inpatient mortality (5.1% 

vs. 8.7%, p


Figure. Changes in fatigue and work productivity scores during 

and after treatment with different anti-HCV regimens. 

Disclosures: 



The following authors have nothing to disclose: Maria Stepanova, Linda Henry, 

Fatema Nader, Sharon L. Hunt 

1461 

Cost-effectiveness of Daclatasvir in Combination with 

Sofosbuvir for the Treatment of Subjects with Genotype 

3 Chronic Hepatitis C Infection in the United States 

Catherine St-Laurent Thibault 1 , Divya Moorjaney 1 , Michael Ganz 1 , 

Bruce E. Sill 2 , Shalini Hede 2 , Yong Yuan 2 , Anupama Kalsekar 2 , 

Boris Gorsh 2 ; 1 Modeling and Simulation, Evidera, St-Laurent, QC, 

Canada; 2 Bristol-Myers Squibb Company, Plainsboro, NJ 

BACKGROUND: An open label phase III trial evaluated the 

efficacy and safety of daclatasvir (DCV) in combination with 

sofosbuvir (SOF) for treatment of subjects with genotype (GT) 

3 hepatitis C virus (HCV). This study evaluated the cost-effectiveness 

of DCV+SOF versus SOF in combination with ribavirin 

(RBV) over a time horizon of 20 years from the payer perspective 

in the United States (US). METHODS: A published Markov 

model was adapted to reflect US baseline demographic 

characteristics, treatment patterns, costs of drug acquisition, 

monitoring, disease management and adverse event management, 

and mortality risks. Clinical inputs were based on the 

ALLY3 trial for DCV+SOF and the VALENCE trial for SOF+RBV. 

The primary cost-effectiveness outcome was the incremental 

cost-utility ratio (ICUR). Life-years, incidence of complications 

(compensated cirrhosis, decompensated cirrhosis, hepatocellular 

carcinoma, liver transplant and liver-related death), 

number of patients achieving sustained virological response 

(SVR) and the total cost per SVR were used as the secondary 

outcomes. Costs (2014 USD) and QALYs were discounted at 

3% per year. An estimated cost was used for DCV based on 

the current standard of care. Deterministic sensitivity analyses 

(DSA) (varying SVR probabilities [+/-10%], health state and 

therapy costs [+/-20%]), probabilistic sensitivity analyses (PSA) 

(with probabilistic sampling of key inputs at each of the 1,000 

iterations using their respective standard error) and scenario 

analyses (evaluating the impact of various time horizons [5, 10 

and 80 years] along with analyses for specific subpopulations 

[treatment-naïve, treatment-experienced, cirrhotic or non-cirrhotic]) 

were conducted to assess the robustness of the results 

to changes in inputs and assumptions. RESULTS: Compared 

with SOF+RBV, DCV+SOF was a dominant treatment in the 

base case as well as in almost all scenarios (i.e., treatment-experienced. 

non-cirrhotic, lower DCV price, 5 and 10 year time 

horizon). In the cirrhotic and treatment naïve population scenarios, 

DCV+SOF was less costly but also slightly less effective 

compared to SOF+R. DSA for the overall population showed 

that ICURs were sensitive to variations in the probability of 

achieving SVR for DCV+SOF and SOF+RBV. PSA for the overall 

population indicated that DCV+SOF yielded an ICUR below 

$50,000 per QALY gained in 78.7% of all iterations compared 

to SOF+RBV. CONCLUSION: DCV+SOF is a dominant 

option compared with SOF+RBV in the US for the overall GT3 

HCV patient population. It may be important to understand the 

real-world SVR rates, as variations in this efficacy parameter 

can have a significant impact on the ICUR. 

Disclosures: 

Bruce E. Sill - Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers 

Squibb 

Yong Yuan - Employment: Bristol Myers Squibb Company 

Anupama Kalsekar - Employment: Bristol Myers Squibb 

Boris Gorsh - Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers 

Squibb 

The following authors have nothing to disclose: Catherine St-Laurent Thibault, 

Divya Moorjaney, Michael Ganz, Shalini Hede 

1462 

The cost-effectiveness of ombitasvir/paritaprevir/ritonavir, 

dasabuvir and ribavirin (3D+R) in patients with 

liver transplantation after genotype 1 hepatitis C virus 

infection 

Sammy Saab 2 , Yuri Sanchez 3 , Caroline Huber 1 , Alice Wang 3 , 

Timothy R. Juday 3 ; 1 Precision Health Economics, Los Angeles, CA; 

2 Pfleger Liver Institute, University of California, Los Angeles, Los 

Angeles, CA; 3 AbbVie, Inc., North Chicago, IL 

Background: Hepatitis C virus (HCV) is the most common indication 

for liver transplantation, and HCV infection recurs in 

over 90% of patients within a month of transplant. Liver fibrosis 

progresses more rapidly after recurrence, and treatment 

options historically have been limited in this population. New 

interferon-free regimens have demonstrated safety and efficacy 

in the post-transplant HCV population; we explore the cost-effectiveness 

of regimens with FDA-approval or Phase III trial 

results in the post-liver transplant setting from a US healthcare 

system perspective. Methods: A cohort of post-liver transplant 

patients with recurrent HCV genotype 1 and fibrosis stage 

F0-F2 was modeled using a two-phase Markov model. In the 

first phase, all patients underwent one of three treatment scenarios: 

no treatment (NT), pegylated interferon and ribavirin 

for 48 weeks (PR48), or ombitasvir/paritaprevir/ritonavir,


dasabuvir and ribavirin for 24 weeks (3D+R). This phase distributed 

patients into post-treatment disease states, including 

sustained virologic response (SVR), based on treatment efficacy. 

The second phase followed the cohort’s post-treatment 

progression through further liver fibrosis and death. Outcome 

measures included lifetime costs (medical and pharmaceutical), 

quality-adjusted life years (QALYs) and incremental cost-effectiveness 

ratios (ICERs) associated with each treatment. SVR 

and adverse event rates were based on phase III clinical trials. 

Transition probabilities, utilities and medical costs were 

extracted from published literature; treatment costs assumed 

100% adherence and reflected wholesale acquisition costs 

from December 2014 Red Book. Results: In a post liver-transplant 

setting, the use of 3D+R had the lowest discounted overall 

costs ($423,585) and highest gain in QALYs (11.3) compared 

to NT ($724,757 cost, 8.25 QALYs) or PR48 ($658,411 cost, 

8.73 QALYs). Use of 3D+R also generated net cost-savings 

for the healthcare system given the lower cost of this regimen 

($169,018) compared to the incremental discounted costs of 

NT ($301,172) and PR48 ($234,826). Conclusions: Compared 

to other regimens for treatment of post-liver transplant 

genotype 1 HCV recurrence, 3D+R is a cost-effective option 

that generates significantly higher quality-adjusted survival and 

lower costs. Use of 3D+R in the post-liver transplant population 

could also generate net cost savings for the healthcare system 

compared to previous standards of care. 

1463 

Gut Microbiota Alterations are an Independent Prognostic 

Factor for 90-day hospitalization in Cirrhosis 

Jasmohan S. Bajaj 1 , Naga Betrapally 2 , Douglas M. Heuman 1 , Melanie 

White 1 , Ariel Unser 1 , Edith A. Gavis 1 , Phillip B. Hylemon 1 , 

Leroy Thacker 1 , Masoumeh Sikaroodi 2 , Patrick M. Gillevet 2 ; 1 VCU 

and McGuire VAMC, Richmond, VA; 2 Microbiome Analysis Center, 

Geoge Mason University, Manassas, VA 

Gut microbiota abnormalities (dysbiosis) worsens with the progression 

of cirrhosis in cross-sectional studies. However the 

impact of fecal dysbiosis on hospitalizations in cirrhotics is 

unclear. Aim: Define the association of dysbiosis with 90-day 

hospitalizations in cirrhosis. Methods: 284 cirrhotics (56 yrs, 

MELD 11, 35% alcohol, 38% prior HE, 30% Diabetes) underwent 

stool multi-tagged sequencing analysis to determine the 

relative microbial family abundances including beneficial commensals 

such as Lachnospiraceae, Ruminococceae & Clostridiales 

XIV. Subjects were followed for 90 days and non-elective 

hospitalizations were noted. Binary logistic regression using 

90-day hospitalizations as the dependent variable with MELD, 

PPI use, HE, alcohol etiology, diabetes, and bacteria significant 

on univariate analyses as predictors was performed. 

Results: Of the 284, 25 were lost to follow-up and 5 had elective 

hospitalizations. 94 (37%) were non-electively hospitalized 

within 90 days (median 35, IQR 21-78 days). The major 

(n=87) hospitalization reason were liver-related, thus no further 

sub-analysis was done(HE=46, Infection=14, Anasarca=13, 

GI bleed=10, others=4). Those who were hospitalized had 

a worse cirrhosis severity, were younger, and higher PPI use 

(Table). Alcoholic etiology was similar (25% vs. 18%, p=0.4). 

Stool microbiota showed a lower relative abundance of Bacteroidaceaeae 

(11% vs 26%, p=0.002), Lachnospiraceae (8% 

vs 14%,p=0.001), Ruminococceae (3% vs. 7%,p


years; HBV/HCV/alcohol/NASH/others = 1/16/9/4/6). 

Data were statistically compared and examined both before 

and 1 month after B-RTO. Results: Preoperative Child-Pugh (CP) 

score was higher in the HE group than in the GV group (8.4 

vs. 6.2 pts., p < 0.01), and both dilatation of splenic vein 

(12.7 vs. 9.0 mm, p < 0.01) and constriction of portal vein 

(9.6 vs. 12.0 mm, p < 0.01) pre-B-RTO were significant in the 

HE group compared with in the GV group. Moreover, while 

splenic venous blood flow in the GV group was all hepatopetal, 

blood flow in 80% of the patients in the HE group was 

hepatofugal before the procedure. Whereas increased portal 

blood flow, elevated portal venous pressure, and expanded 

portal vein diameter following B-RTO were statistically significant 

in both groups, an improvement in CP score was more 

marked in the HE group (HE: 8.4 to 7.0 pts., p < 0.01; GV: 

6.2 to 5.9 pts., p < 0.05). Splenic vein diameter was significantly 

reduced in the HE group (12.7 to 9.9 mm, p < 0.01), 

meanwhile it was markedly increased in the GV group (8.9 to 

9.4 mm, p = 0.07), suggesting that splenic venous blood flow 

in the HE group changed from hepatofugal to hepatopetal after 

B-RTO. The decrease in blood ammonia levels in response to 

shunt occlusion was also significant in both groups, but it was 

statistically more marked in the HE group (HE: 161.5 to 95.2 

μg/dL, p < 0.01; GV: 90.8 to 69.4 μg/dL, p < 0.05). This 

effect lasted at least until 6 months after the procedure (before: 

161.5 μg/dL, after 1 month: 95.2 μg/dL, after 3 months: 

69.7 μg/dL, after 6 months: 64.6 μg/dL). Conclusions: Much 

like the dramatic effect on GV, B-RTO has an extremely strong 

and long-lasting effect on refractory HE. Moreover, compared 

with those treated for GV, patients treated for HE exhibit highly 

characteristic “hemodynamics in the portal–splenic venous system” 

both before and after the B-RTO procedure, which may 

lead to markedly improved hepatic function including ammonia 

metabolism. 

Disclosures: 

The following authors have nothing to disclose: Tatsuro Nishimura, Tsuyoshi 

Ishikawa, Ryo Sasaki, Yuki Aibe, Shogo Shiratsuki, Kazuhito Matsunaga, Takuya 

Iwamoto, Taro Takami, Isao Sakaida 

1465 

Lactulose treatment improves cognition, quality of life 

and intestinal microbiota in minimal hepatic encephalopathy 

patients: results from a multi-center, randomized, 

controlled trial 

Jiyao Wang 1 , Jiang-bin Wang 2 , Jia Shang 3 , Xin-Min Zhou 4 , Xiao- 

Lin Guo 5 , Xuan Zhu 6 , Li-Na Meng 7 , Hai-Xing Jiang 8 , Yu-Qiang 

Mi 9 , Jian-Ming Xu 10 , Jin-Hui Yang 11 ; 1 GI & Hepatology, Zhong 

Shan Hospital,Fudan University, Shanghai, China; 2 Gastroenterology, 

China-Japan Union Hospital, Jilin University, Changchun, 

China; 3 Infectious Disease, Departmentof Infectious Diseases, 

Henan Provincial People’s Hospital, Zhengzhou, China; 4 Digestive 

Diseases,, Xijing Hospital, the Fourth Military Medical University, 

Xi’an, China; 5 Digestive Diseases, The First Hospital of Jilin University, 

Changchun, China; 6 Digestive Diease, The first Affiliated 

Hospital of Nanchang University, Nanchang, China; 7 Digestive 

Diseases, Zhejiang Provincial Hospital of Tranditional Chinese 

Medicine, Hangzhou, China; 8 Digestive Diseases, The First AffiliatedHospital 

of Guangxi Medical University,, Nanning, China; 

9 Hepatology, Tianjin Second People’s Hospital, Tianjin, China; 

10 Digestive Diseases, The First Affiliated Hospital of Anhui Medical 

University, Hefei, China; 11 Hepatology, The second Affiliated Hospital 

of Kunming Medical University, Kunming, China 

Aims As we investigated recently, minimal hepatic encephalopathy 

(MHE) is high prevalent in China (39.8 %) and those 

patients with low quality of life (QoL). Following previous findings, 

this study is to assess the efficacy and safety of lactulose 

in treatment of MHE in aspect of cognitive function, QoL, and 

intestinal microbiota. Methods It is a nationwide, multi-centered, 

randomized controlled trial. Patients with liver cirrhosis 

were screened by number connection test A (NCT-A) and digit 

symbol test (DST) both positive to determine MHE diagnose. 

These MHE patients were 2: 1 randomized to receive lactulose 

30-60ml/twice per day (Gp-L) or no therapy (Gp-NL). At month 

2, the psychometric tests (NCT-A and DST), the QoL questionnaire, 

the Child-Pugh score, the ammonia and the intestinal 

flora (metastats, QIIME) were reassessed. The primary endpoint 

was the recovery rate defined as the normalization of 

at least one of the psychometric tests. Results Of 575 patients 

screened, 98 (17.04%) met the inclusion and exclusion criteria 

(Gp-L n=67, Gp-NL n=31). In all, 11(11.22%) patients were 

lost to follow-up (Gp-L n=8, Gp-NL n=3). At month 2, the MHE 

recovery rate in Gp-L (41/59, 69.49%) is significantly higher 

than in Gp-NL (6/28, 21.43%) (p=0.0000). The QoL in Gp-L 

was significantly improved from baseline (p=0.0000) in all 5 

aspects of physical function, psychological well-being, relief of 

symptoms, social function and self-evaluation. The improved 

score of QoL questionnaire in Gp-L (12.63±15.10) was larger 

than Gp-NL (6.36±12.16) (p=0.0523). Two-month lactulose 

treatment significantly improved the severity of liver cirrhosis 

(assessed by Child-Pugh score) (p=0.0013) decreased ammonia 

level (delta= -63.16±3.60) (p=0.0000) from baseline. 

Furthermore, the intestinal microbiota changed in both autochthnous 

taxa and non-autochthnous taxa in Gp-L, an increase of 

Bifidobacteriaceae (2.9% to 5.8%), Lactobacillaceae (4.3% to 

6%), Ruminococcaceae Ruminococcus (1.2% to 2.2%), and a 

decrease of Rikenellaceae Alistipes (4.5% to 3.3%) were seen 

at the phylum/order level. All patients were tolerated lactulose, 

no severe side effects was reported. Conclusion Lactulose is 

effective for the recovery of MHE in aspect of cognition, QoL 

in patients with cirrhosis, which may be associated with the 

microbial changes. Regard this, the prophylactic therapy by 

lactulose is suggested for MHE patients. Clinical trial nr: ChiC- 

TR-TRC-12002342 

Disclosures: 

The following authors have nothing to disclose: Jiyao Wang, Jiang-bin Wang, 

Jia Shang, Xin-Min Zhou, Xiao-Lin Guo, Xuan Zhu, Li-Na Meng, Hai-Xing Jiang, 

Yu-Qiang Mi, Jian-Ming Xu, Jin-Hui Yang 

1466 

Transient elastography related cut off parameters in 

patients with advanced chronic liver disease independently 

predict decompensation and mortality but 

lack prediction of varices. 

Rafael Paternostro, Monika Ferlitsch, Alexandra Etschmaier, 

Remy Schwarzer, Thomas Reiberger, Mattias Mandorfer, Michael 

Trauner, Markus Peck-Radosavljevic, Arnulf Ferlitsch; Vienna 

Hepatic Hemodynamic Lab, Div. of Gastroenterology and Hepatology, 

Internal Medicine III, Medical University of Vienna, Vienna, 

Austria 

Background & Aims: Transient elastography (TE) is a validated 

tool to stage liver fibrosis and identify patients with advanced 

chronic liver disease (ACLD) by liver stiffness >10kPa. The 

Baveno VI consensus conference on portal hypertension 

proposed to select a TE value of 20kPa to stratify patients 

with ACLD. Aim of this study was to evaluate the predictive 

value of TE regarding (i) decompensation and (ii) mortality. 

Methods: 227 patients with chronic liver diseases were evaluated 

by TE. 55 patients had liver stiffness


included. 132 (85%) underwent additional HVPG measurements. 

Hepatic decompensation (ascites, HE, jaundice) and 

mortality was recorded during follow-up. Results: Baseline 

characteristics: age: 52±11years, 73% male, main etiologies: 

47.7% viral, 31% ALD, 21.3% others. Median TE values were 

29.1kPa (11.3-75). Mean HVPG was:15±6.5mmHg. Median 

follow up was 967 days(89.2-1609.2). In total 56/155 

patients decompensated or died during follow up. Hepatic 

decompensation or death occurred significantly more often in 

patients with TE>20kPa: 49/106 (46.2%) than in patients with 

TE20kPa is an independent risk factor 

for decompensation or mortality in ACLD patients. Considering 

that numbers of ACLD patients with TE150G/L are low, still around 50% showed varices – not 

supporting the Baveno VI recommendation to avoid screening 

endoscopy in this constellation. The 20kPa stratification strategy 

should be prospectively evaluated in larger cohorts. 

Disclosures: 













The following authors have nothing to disclose: Rafael Paternostro, Monika Ferlitsch, 

Alexandra Etschmaier, Remy Schwarzer, Arnulf Ferlitsch 

1467 

Development of an Electronic Diary (E-Diary) for Caregivers 

(CG) of Patients with Overt Hepatic Encephalopathy 

(OHE) 

R Todd Frederick 1 , Marwan Ghabril 2 , Karin Coyne 3 , Mary Kay 

Margolis 3,4 , Michael Santoro 5 , Dion F. Coakley 5 , Masoud Mokhtarani 

5 , Bruce F. Scharschmidt 5 , Marzena Jurek 5 ; 1 California Pacific 

Medical Center, San Francisco, CA; 2 Indiana University, Indianapolis, 

IN; 3 Evidera, Bethesda, MD; 4 Patient-Centered Outcomes 

Research Institute (PCORI), Washington, DC; 5 Horizon Therapeutics 

(formerly Hyperion Therapeutics, Inc.), Brisbane, CA 

Care of patients with OHE requires education and involvement 

of the CG, as most episodes occur outside of the hospital or 

clinic. However, some CG may try to manage the episodes 

without informing or receiving input from the medical provider. 

Moreover, lack of systematic patient evaluation and/or communication 

with the provider by CG may hinder identification of 

OHE episodes in clinical practice and represents a challenge 

in the design and interpretation of OHE endpoints in clinical 

trials. Study Design: A 3 part approach was used to develop a 

daily e-diary for CG of OHE patients. Part 1 was conducted at 

2 clinical sites and included concept elicitation, i.e. interviews 

with 12 CG who had cared for a patient with OHE within the 

last three months, to determine the signs and symptoms CG use 

to identify OHE. In Part 2, a daily e-diary for CG was developed 

that included questions with branching logic and skip 

patterns for standardized feedback, automatic reminders for 

completion, and real time alerts sent to the site if CG answers 

were suggestive of OHE. This e-diary was evaluated for comprehensiveness 

and understandability by 10 CG in semi-structured 

interviews. Part 3 was a 3-day field test among 10 CG 

to assess real-world usability on their own devices. The final 

e-diary was translated and culturally adapted with CG input 

into 16 languages. Results: Part 1 CG collectively described 

33 different HE-related symptoms experienced by the patients 

they cared for. Forgetfulness/confusion and sleep difficulties 

(e.g., day-night inversion) were reported by all CG; most also 

described speech problems (difficulty finding words [92%] or 

repeating words or phrases [50%]). No one symptom emerged 

as most important and most CG (83%) reported that they did 

not contact a physician unless there was progression or non-resolution 

of OHE symptoms. Part 2 interviews identified 7 items 

(speech difficulties, unusual behavior, forgetfulness/confusion, 

orientation, and level of consciousness), which adequately and 

comprehensively captured what CG observe and monitor at 

home and that the e-format was user friendly. Part 3 confirmed 

the usability of the e-diary on the CG’ personal devices, including 

web-enabled smart phones (iPhone, Android), laptop, and 

desktop computers. Conclusion: A comprehensive, easy to 

use CG e-diary was developed for use in clinical trials of OHE 

and in clinical practice. Real time communication with health 

care providers and standardized evaluation of manifestations 

of OHE by CG will improve documentation of endpoints in 

trials of OHE and may lead to earlier medical intervention and 

improved patient outcomes in clinical practice. 

Disclosures: 

R Todd Frederick - Advisory Committees or Review Panels: Gilead, Bristol Myers 

Squibb, Salix, Vital Therapies, Hyperion, AbbVie 

Marwan Ghabril - Grant/Research Support: Salix 

Karin Coyne - Consulting: Evidera 

Michael Santoro - Employment: Horizon Therapeutics, Inc. 

Dion F. Coakley - Employment: Horizon Therapeutics 

Masoud Mokhtarani - Employment: Hyperion; Stock Shareholder: Hyperion 

Marzena Jurek - Employment: Horizon Therapeutics, Inc. 

The following authors have nothing to disclose: Mary Kay Margolis, Bruce F. 

Scharschmidt 

1468 

Muscle mass in liver transplant candidates: From the Fitness, 

Life Enhancement, and Exercise in Transplantation 

(FLEXIT) Consortium 

Jennifer C. Lai 1 , Elizabeth J. Carey 2 , Connie W. Wang 1 , Srinivasan 

Dasarathy 3 , Michael A. Dunn 4 ; 1 UCSF, San Francisco, CA; 

2 Mayo Clinic Arizona, Scottsdale, AZ; 3 Cleveland Clinic, Cleveland, 

OH; 4 University of Pittsburgh, Pittsburgh, PA 

Background: Sarcopenia, frailty, and deconditioning are distinct 

terms for anatomic and functional disturbances related to 

muscle wasting, a frequent and often lethal manifestation of 

advanced cirrhosis. We hypothesized that sarcopenia measured 

on cross sectional imaging as an indicator of muscle 

wasting could be consistently evaluated as a common parameter, 

and that its clinical significance in liver transplant candidates 

from multiple centers would confirm earlier single-center


reports. Methods: Adult liver transplant candidates listed in 

2012 with an abdominal CT scan within 3 months of listing 

were included. Our 4 centers executed a data use agreement 

and agreed on common definitions of clinical variables, key 

parameters to measure muscle mass, and outcomes. Abdominal 

skeletal muscle area was measured as the sum of cross-sectional 

areas of psoas, paraspinal, and abdominal wall muscles 

at L3, normalized for height (SMI, cm 2 /m 2 ). Associations 

between SMI and baseline characteristics were evaluated 

by Pearsons correlations, Wilcoxon, or Kruskal-Wallis tests. 

Adjusted competing risks analysis evaluated association of SMI 

with death/delisting for being too sick for LT. Results: Included 

were 224 subjects: 25% female, 54% non-Hispanic White, 

54% HCV, 14% ETOH, 11% NASH, 46% HCC, median age 

59y, median BMI 28. Disease-related characteristics: median 

MELD 14, median albumin 3.1, 49% with ascites. Median 

(IQR) total abdominal skeletal muscle index was 46 cm 2 /m 2 

(36-55). SMI had low correlation with MELD (r=-0.27;p


analysis identified altered levels of metabolites associated with 

bile acids, heme metabolism, sphingosine and lipids in HPS 

relative to controls. Levels of 6 of 8 primary bile acids and 

only 2 of 19 secondary bile acids were increased in HPS relative 

to control (others unchanged) suggesting altered excretion 

and gut microbial metabolism that may affect the vasculature. 

Products of heme metabolism including bilirubin, biliverdin and 

urobilinogen were also increased in HPS suggesting enhanced 

hemoxygenase driven heme metabolism and/or impaired 

clearance which may increase carbon monoxide production. 

Levels of sphingosine metabolites, potent regulators of endothelin 

and nitric oxide synthase signaling, were also elevated in 

HPS. Finally, global increases in fatty acid levels with reduced 

monoglycerol production were found in HPS suggesting 

enhanced fatty acid release or decreased clearance. Random 

forest analysis showed that selected metabolites distinguished 

HPS from controls with 71% accuracy. Conclusion: Human HPS 

has plasma metabolic signatures consistent with alterations 

in sphingosine, bile acid, heme and fatty acid metabolism, 

suggesting novel mechanistic pathways and possible links to 

experimental HPS. Validation in larger cohorts is required. 

Disclosures: 

Michael B. Fallon - Grant/Research Support: Bayer/Onyx, Eaisi, Gilead, Grifolis 

The following authors have nothing to disclose: Michael J. Krowka, Kimberly A. 

Forde, Karen Krok, MAMTA PATEL, Grace Lin, Jae K. Oh, Carl Mottram, Paul D. 

Scanlon, Sachin Batra, David S. Goldberg, Steven M. Kawut 

and of BMI 27.12 ± 5.24. The frequency of malnutrition was 

14.7% for BIA, 25% for BIVA, 32% for MAMC, 13% for TSF, 

and 14 for % BMIa, In the Rxc graph only patients with clinical 

ascites displayed fluid retention by BIVA (55.1% vs 44.9% 

without ascites, p=0.005). Also, higher stages of Child-Pugh 

were significantly associated with a higher rate of malnutrition 

and fluid retention (data not shown). Kaplan-Meier curves disclosed 

a higher mortality in the malnourished group identified 

by BIVA (39.7% vs. 21.4% in non malnourished, p=


were significant. On multi-variate analysis, recent alcohol (OR 

4.0, p=0.003) and illegal turns (OR 1.4, p=0.005) remained 

significant predictors of real-life crashes. Conclusion: In this 

largest-studied cirrhosis cohort, MHE subjects were likely to 

have worse simulator and real-life crash rates that remains 

stable over 1 year. Only MHE patients who performed poorly 

on navigation and had quit alcohol within 3 years were likely 

to be unsafe drivers. 

Summarized of results 

Table shows summarized of results from driving simulation, real 

life traffic history and 6-month follow up in 163 patients. 

Disclosures: 

Richard K. Sterling - Advisory Committees or Review Panels: Merck, Vertex, Salix, 

Bayer, BMS, Abbott, Gilead, Baxter, Jansen; Grant/Research Support: Merck, 

Roche/Genentech, Pfizer, Gilead, Boehringer Ingelheim, Bayer, BMS, Abbott 


Inc.; Consulting: NPS Pharmaceuticals Inc. 






Douglas M. Heuman - Consulting: Bayer, Grifols, Genzyme; Grant/Research 

Support: Exilixis, Novartis, Bayer, Bristol Myers Squibb, Scynexis, Ocera, Mannkind, 

Salix, Globeimmune, Roche, SciClone, Wyeth, Otsuka, Ikaria, UCB, Celgene, 

Centocor, Millenium, Osiris, AbbVie, Gilead; Speaking and Teaching: 

Otsuka, Astellas 

Jasmohan S. Bajaj - Advisory Committees or Review Panels: Salix, Merz, otsuka, 

ocera, grifols, american college of gastroenterology; Grant/Research Support: 

salix, otsuka, grifols 

The following authors have nothing to disclose: Mette M. Lauridsen, Vishwadeep 

Ahluwalia, Ariel Unser, Melanie White, R. Todd Stravitz, Scott C. Matherly, 

Velimir A. Luketic, Mohammad S. Siddiqui, Michael Fuchs 

1473 

Randomized Controlled Trial on Efficacy of Nutritional 

Therapy on Cognitive Functions and Health Related 

Quality of Life in Patients of Liver Cirrhosis with Minimal 

Hepatic Encephalopathy 

Sudhir Maharshi, Barjesh C. Sharma, Sanjeev Sachdeva, Siddharth 

Srivastava; Gastroenterology, G B Pant Institute of Post 

graduate Medical Education and Research, New delhi, India 

Background & Aims: Minimal hepatic encephalopathy impairs 

health related quality of life (HRQOL), predicts development of 

overt hepatic encephalopathy and associated with poor prognosis. 

There is no study on nutritional management in patients 

with minimal hepatic encephalopathy. We assessed the effects 

of nutritional therapy on cognitive functions and HRQOL in 

patients of cirrhosis with minimal hepatic encephalopathy. 

Methods: A randomized controlled trial conducted in a tertiary 

care setting on patients of cirrhosis with minimal hepatic 

encephalopathy who were randomized to nutritional therapy 

(group A: 30-35 kcal/kg/day and 1 . 0-1 . 5 gram of vegetable 

protein/kg/day) and no nutritional therapy (group B: diet as 

patients were taking before) for 6 months. Minimal hepatic 

encephalopathy was diagnosed based on psychometry hepatic 

encephalopathy score (PHES). HRQOL was assessed by sickness 

impact profile (SIP) questionnaire. Primary endpoints were 

improvement or worsening in minimal hepatic encephalopathy 

and improvement in HRQOL. Results: 120 patients were 

randomized to group-A (n = 60, age 42 . 1±10 . 3 yr, 48 men) 

and group-B (n = 60, age 42 . 4±9 . 6 yr, 47 men). There was 

no significant difference in baseline characteristics between 

the two groups. Baseline PHES (-8 . 12±1 . 32 vs -8 . 53±1 . 38; 

p=0 . 08) and SIP score (14 . 25±5.8 vs 15 . 44±5 . 03; p=0 . 85) 

were comparable in both the groups. Reversal of minimal 

hepatic encephalopathy was higher in group A (71 . 1% vs 

22 . 8%; p=0 . 001). Improvement in PHES (ΔPHES 3 . 86±3 . 58 

vs 0 . 52±4 . 09; p=0 . 001) and HRQOL (Δ SIP 3 . 24±3 . 63 vs 

0 . 54±3 . 58; p=0 . 001) were also higher in group A compared 

to group B. Overt hepatic encephalopathy developed in 10% 

of patients in group A vs 21 . 7% in group B (P=0 . 04). Conclusions: 

Nutritional therapy is effective in treatment of minimal 

hepatic encephalopathy and associated with improvement in 

HRQOL. 

Disclosures: 

The following authors have nothing to disclose: Sudhir Maharshi, Barjesh C. 

Sharma, Sanjeev Sachdeva, Siddharth Srivastava 

1474 

Usefulness of Balloon-Occluded Retrograde Obliteration 

(B-RTO) as a Therapeutic Procedure to Improve Liver 

Function in Cirrhotic Patients with Porto-Systemic Shunts 

Manabu Nakazawa, Yukinori Imai, Satsuki Ando, Kayoko Sugawara, 

Mie Inao, Nobuaki Nakayama, Satoshi Mochida; Gastroenterology 

& Hepatology, Saitama medical university hospital, 

Saitama, Japan 

Aim: Porto-systemic shunts cause refractory hepatic encephalopathy 

in patients with liver cirrhosis. The shunts also contribute 

to derange liver function through reduction of blood flow in 

the portal vein. The usefulness of B-RTO as a therapeutic procedure 

for improvement of liver function as well as attenuation of 

refractory hepatic encephalopathy was evaluated in cirrhotic 

patients. Methods: The subjects were 34 cirrhotic patients with 

porto-systemic shunts showing refractory hepatic encephalopathy. 

The extent of liver dysfunction classified according to the 

Child-Pugh classification were grade A, B and C in 3, 18 and 

13 patients, respectively. A balloon catheter was inserted into 

the shunts followed by injection of 5% ethanolamine oleate 

through the catheter under balloon inflation. The balloon was 

kept inflation for between 6 and 48 hours depending on 

sizes of the shunts. Results: Drainage vessels of the targeted 

shunts were the splenorenal, mesocaval, paraumbilical, azygos 

and splenorenal plus azygos veins in 20, 4, 6, 2 and 

2 patients, respectively. B-RTO was successfully done in 29 

patients (85%), and hepatic encephalopathy did not recur in 

24 (83%) patients. In patients receiving successful B-RTO procedures, 

HH15, LHL15 and LU15 values assessed by Tc-99m 

GSA scintigraphy were not different between at baseline and 

1 months after the procedures, while doppler ultrasound examinations 

revealed that blood flows in the portal vein (cm 3 /sec; 

mean±SD) increased significantly (from 2.2±0.9 to 8.4±2.4, 

p


therapeutic procedure to improve liver function as well as to 

attenuate hepatic encephalopathy through increase of portal 

venous flows in cirrhotic patients with porto-systemic shunts. 

Disclosures: 



The following authors have nothing to disclose: Manabu Nakazawa, Yukinori 

Imai, Satsuki Ando, Kayoko Sugawara, Mie Inao, Nobuaki Nakayama 

1475 

Six-week administration of lactulose in patients with cirrhosis 

does not alter the gut bacterial flora 

Amit Goel 1 , Aditya N. Sarangi 2 , Ankur Singh 1 , S. Avani 1 , Rakesh 

Aggarwal 1,2 ; 1 Gastroenterology, Sanjay Gandhi Postgraduate 

Institute of Medical Sciences, Lucknow, India; 2 Biomedical informatics 

Center, Sanjay Gandhi Postgraduate Institute of Medical 

Sciences, Lucknow, India 

Background: Benefit of lactulose in hepatic encephalopathy 

(HE) may be mediated, at least partially, by alterations in gut 

flora. However, data on change in gut flora post-lactulose are 

sparse. We therefore studied gut microbial composition in 

patients with liver cirrhosis (LC) and the effect of 6 weeks of 

lactulose use on it, using next-generation sequencing. Methods: 

30 patients with LC (male 22; median [range] age: 42 [29-65] 

y; body mass index 22.7 [17.3-32.3] Kg/m 2 ) of any cause 

or severity but no prior history of HE, and 18 healthy controls 

(HC; male 14; 45 [24-67] y; 23.3 [20.0-25.0] Kg/m 2 ) were 

enrolled. Persons with gastrointestinal symptoms, or recent (preceding 

6 weeks) use of lactulose or another drug affecting 

gut motility or flora were excluded. A morning stool specimen 

was collected from each subject; 19 patients also provided 

a follow-up specimen after 6 weeks of lactulose intake. A 

cDNA library for V3 region of bacterial 16S rRNA from each 

stool was subjected to paired-end Illumina sequencing, and 

abundances of various bacterial operational taxonomic units 

(OTUs) were determined. Relationship of specimens with each 

other was studied using principal co-ordinate analysis (PCoA). 

Abundances of various OTUs, and indices of α and β diversity 

were compared between groups using t-test with Bonferroni 

correction; data before and after lactulose were compared 

using tests for paired data. Results: The 67 specimens yielded a 

median (range) of 504,182 (171,799-1,267,206) high-quality 

reads. In PCoA, gut flora from LC clustered separately from 

HC and showed lower diversity (Chao 1 index: 1975 vs 2802; 

observed species: 1082 vs 1485; Shannon index: 4.8 vs 5.6; 

p=0.0001 each). Median abundances of major phyla (Bacteroidetes 

[75.2% vs 67.2%], Firmicutes [17.5% vs 18.4%] 

and Proteobacteria [6.6% vs 8.3%]) were similar in LC and 

HC; however, Lentisphaerae [0% vs 0.01%], Cyanobacteria 

[0.01% vs 0.52%] and Tenericutes [0.01% vs 0.05%] were 

less abundant in LC than HC (p0.005% 

of all the bacterial sequences had comparable abundances 

before and after lactulose. The pre- and post-lactulose specimens 

showed no separation on PCoA, and had similar α diversity 

indices (Chao 1: 2755 vs 2788; observed species 1732 

vs 1761; Shannon index: 4.9 vs 4.9; all p=ns). Conclusion: 

Gut flora in LC differs from healthy persons. However, administration 

of lactulose for 6 weeks did not produce a discernible 

change in gut flora in patients with LC, suggesting that alteration 

in gut flora may play a minor role in improving HE. 

Disclosures: 

The following authors have nothing to disclose: Amit Goel, Aditya N. Sarangi, 

Ankur Singh, S. Avani, Rakesh Aggarwal 

1476 

Erectile Dysfunction in patients with cirrhosis 

Sergio Maimone 1 , Giovanni Oliva 1,2 , Antonino Di Benedetto 2 , 

Roberto Filomia 1,2 , Gaia Caccamo 1 , Carlo Saitta 1 , Irene Cacciola 

1,2 , Giovanni Squadrito 1,3 , Giovanni Raimondo 1,2 ; 1 Division 

of Clinical and Molecular Hepatology, Universitary Hospital of 

Messina, Messina, Italy; 2 Department of Clinical and Experimental 

Medicine, University Hospital of Messina, Messina, Italy; 3 Department 

of Human Pathology, University Hospital of Messina, Messina, 

Italy 

BACKGROUND AND AIMS - Erectile dysfunction (ED) is a clinical 

disorder frequently observed in men suffering from several 

chronic diseases as diabetes, obesity, metabolic syndrome, 

cardiovascular diseases. There is little information, however, 

about ED in patients with advanced chronic liver disease. Aims 

of our study were to evaluate the prevalence of ED in cirrhotic 

patients and to investigate clinical and biochemical factors 

possibly associated with ED occurrence in these patients. 

PATIENTS AND METHODS - We prospectively analyzed 147 

consecutive cirrhotic patients (mean age 61.5± 15.5 years) 99 

of whom were in class A and 48 in class B of Child-Pugh (CP). 

Physical examination, electrocardiography, biochemistry as 

well as serum levels of thyroid stimulating hormone (TSH), total 

testosterone (TT), free testosterone (FTT), sex hormon binding 

globulin (SHBG), and prolattin (PRL) were evaluated in each 

patient. Known ED risk factors such as smoking habit, alcohol 

abuse, diabetes, cardiovascular diseases (i.e., hypertensive 

cardiovascular disease, ischemic heart disease), depression, 

hypogonadism, treatment with diuretics and beta blockers 

were evaluated as well. All patients completed the following 

questionnaires: International Index of Erectile Function (IIEF) 

scale (that classifies ED into 4 categories: absent, mild, moderate 

and severe), Self-rating Anxiety Scale (SAS) (that evaluates 

grades of depression), and ANDROTEST (that assesses 

the presence of hypogonadism jointly to serum level of FTT). 

Data processing and statistical analysis were performed by 

SPSS software. RESULTS - ED was revealed in 86/147 (58.5%) 

patients, 31 (36%) of whom had mild, 7 (8.1%) moderate and 

48 (55.9%) severe ED. No statistically significant differences 

were found when patients with and those without ED were 

compared for age, smoking habit, alcohol consumption, ethiology 

of liver disease, concomitant arterial hypertension, therapy 

with diuretics and beta blockers, depression, thyroid and sexual 

hormone profiles. In analogy, SAS test and ANDROTEST 

provided similar results in the two groups. On the contrary, 

at univariate logistic regression analysis ED was significantly 

associated with CP class B (p=0.04), presence of diabetes 

(p=0.04), and cardiovascular diseases (p=0.01). However, 

only presence of cardiovascular disease maintained the significant 

association with ED occurrence at the multivariate logistic 

regression analysis (OR 3.7, CI 95%1.06-13.44, p=0.03). 

CONCLUSIONS - Erectile dysfunction affects the majority of 

patients with cirrhosis and it is significantly associated with 

cardiovascular diseases in these patients. 

Disclosures: 



The following authors have nothing to disclose: Sergio Maimone, Giovanni 

Oliva, Antonino Di Benedetto, Roberto Filomia, Gaia Caccamo, Carlo Saitta, 

Irene Cacciola, Giovanni Squadrito


1477 

Perceptions of the ‘cirrhotic coagulopathy’ influence low 

yield testing for cerebral hemorrhage in patients with 

cirrhosis and hepatic encephalopathy: an international, 

multi-specialty survey 

Elliot B. Tapper 1 , Laura Mazer 2 ; 1 Gastroenterology, Beth Israel 

Deaconess Medical Center, Boston, MA; 2 Surgery, Beth Israel Deaconess 


Patients with cirrhosis and hepatic encephalopathy (HE) are 

often evaluated for intracranial hemorrhage (ICH) even in the 

absence of focal neurologic findings or trauma due to perceptions 

of a tendency toward bleeding - the so-called ‘cirrhotic 

coagulopathy.’. There is limited data on the prevalence and 

clinical impact of such perceptions. Methods: We conducted 

a prospective survey of physicians in internal medicine (IM), 

emergency medicine (EM), general surgery (GS) and gastroenterology 

(GI) at 35 centers in 5 countries. All respondents 

were presented with a standard HE patient presentation (somnolence, 

no focal neurologic findings, no evidence of a fall/ 

trauma). Subjects rated their likelihood to order computed 

tomography (CT) of the head to exclude ICH on a 7 point 

scale from -3 to +3.They were asked to re-rate their likelihood 

after seeing the patient’s labs (including INR 2.4 and platelet 

count 59) and again after seeing the results of a study 

demonstrating the low yield of CT for this presentation (CGH 

2015;13(1):165). Results: 1033 physicians responded, 86% 

from the US. The proportions of IM, ED, GS and GI were 55%, 

29%, 7% and 9%. The proportion of attendings, fellows and 

residents were 42%, 7% and 49%. The baseline tendency to 

order a CT was lowest among GI attendings, median -2 interquartile 

range (IQR) (-2 – 0.75) and highest among EM physicians, 

2 IQR (1-3). At baseline 56.0% of respondents were 

likely to order a head CT. All groups (by specialty, level of 

training and familiarity with cirrhosis) were significantly more 

likely (p < 0.001 for all and to 0.03 for attending GI) to order 

a head CT after seeing the labs. Of the subjects unlikely to 

order a head CT at baseline, 114 (25.4%) would order after 

the labs. All groups were less likely to order head CTs after 

seeing the research data (mean difference -1.51 95% CI (-1.61 

- -1.41), p < 0.0001). Of the 651 likely to order a head CT 

after the labs, 313 (48.1%) were dissuaded by the published 

yield data. 61.2% of IM physicians interested in head CTs 

were most likely to report disinterest after exposure to the data 

compared to 31.3% of EM physicians, p = 0.0002. Conclusions: 

Misperceptions regarding the coagulopathy of cirrhosis 

are highly prevalent and affect clinical decision-making, but 

are amenable to education. 

Disclosures: 

The following authors have nothing to disclose: Elliot B. Tapper, Laura Mazer 

1478 

Prospective Multicenter Observational Study of Overt 

Hepatic Encephalopathy (OHE): Nomenclature and Disease 

Burden 

Charles S. Landis 1 , Marwan Ghabril 2 , Vinod K. Rustgi 3 , Adrian 

M. Di Bisceglie 4 , Benedict Maliakkal 5 , Don C. Rockey 6 , John M. 

Vierling 7 , Richard Rowell 8 , Michael Santoro 8 , Aimee Enriquez 8 , 

Marzena Jurek 8 , Masoud Mokhtarani 8 , Dion F. Coakley 8 , Bruce F. 

Scharschmidt 8 ; 1 University of Washington, Seattle, WA; 2 Indiana 

University, Indianapolis, IN; 3 University of Pittsburgh, Pittsburgh, 

PA; 4 St Louis University, St Louis, MO; 5 University of Rochester, 

Rochester, NY; 6 Medical University of South Carolina, Charleston, 

SC; 7 Baylor College of Medicine, Houston, TX; 8 Horizon Therapeutics 

(formerly Hyperion Therapeutics, Inc.), Brisbane, CA 

Overview: OHE, a complication of advanced liver disease, is 

devastating to patients and families, burdensome to society, 

and challenging with respect to both diagnosis and nomenclature, 

evidenced by recent ISHEN and AASLD/EASL guidelines. 

However, there is little prospectively collected data regarding 

either the presenting manifestations of OHE or its associated 

economic burden, as reflected, for example, by frequency of 

recurrence and hospitalizations. Study Design: Surveys pertaining 

to OHE were sent to over 200 centers in 23 countries; 

176 centers (67% university and 29% public), which followed 

a median of ~40 patients with OHE responded. A subset of 

centers, which followed a median of ~100 OHE patients was 

invited to participate in an observational study. To be eligible 

for enrollment, patients were required to have a clinical diagnosis 

of cirrhosis and at least 1 episode of OHE within 30 

days prior to consent (qualifying OHE). Once enrolled, patients 

were followed to capture OHE recurrence (on-study OHE). Clinical 

manifestations of both qualifying and on-study OHE episodes 

as recorded in the patients’ charts were noted. Results: 

269 patients with a qualifying OHE event were enrolled at 31 

sites in North America and Europe; data from 260 patients 

who were followed for up to 8 months (mean=73 days) were 

available for this interim analysis (median age=61; median 

time since OHE diagnosis=1.1 years, 61% male, 87% Caucasian). 

The most frequently documented clinical manifestations 

of both qualifying and on-study OHE episodes included confusion 

(73%), change in mental status (56%), disorientation 

(46%), lethargy (44%) and asterixis (44%). West Haven grade 

was recorded in only 27% of all OHE episodes. Fifty seven 

patients had 101 on-study OHE episodes. The median time 

for occurrence of 2 nd , 3 rd and 4 th OHE episodes was 28, 19 

and 11 days, respectively. Of 87 and 47 patients followed for 

at least 3 or 4 months, 32% and 34% experienced a mean 

(SD) of 0.7 (1.5) or 0.9 (1.8) on-study OHE episodes, respectively. 

57% of qualifying and 84% of on-study OHE episodes 

occurred while patients were on rifaximin; 65% of qualifying 

and 91% of on-study OHE episodes were associated with 

hospitalization. Conclusions: Among patients enrolled in this 

multi-center and multi-national observational study, clinical disorientation, 

level of consciousness and asterixis were the most 

commonly documented manifestations of OHE, whereas West 

Haven was infrequently recorded. Despite the availability of 

rifaximin, OHE, as reflected by frequent recurrence and hospitalizations, 

continues to represent a major unmet clinical need 

and societal burden. 

Disclosures: 


Marwan Ghabril - Grant/Research Support: Salix 

Vinod K. Rustgi - Advisory Committees or Review Panels: Abbvie, BMS, Merck; 

Grant/Research Support: Gilead 


Novartis, Bayer, BTG; Grant/Research Support: Gilead, AbbVie




Don C. Rockey - Grant/Research Support: Gilead, Actelion, Hyperion 









Michael Santoro - Employment: Horizon Therapeutics, Inc. 

Aimee Enriquez - Employment: Horizon Pharma 




The following authors have nothing to disclose: Richard Rowell, Bruce F. 

Scharschmidt 

1479 

Sarcopenia predicts moratility after transjugular intrahepatic 

portosystemic shunt placement 

Joshua Anderson 1,3 , Alagappan A. Annamalai 1 , Joseph L. Robinson 

1 , Tram T. Tran 1 , Walid S. Ayoub 1 , Jonah Hirschbein 2 , Georgios 

Voidonikolas, 1 , Irene K. Kim 1 , Vladamir Donchev 1 , Nicholas 

N. Nissen 1 , Andrew S. Klein 1 , Vinay Sundaram 1 ; 1 Cedars-Sinai 

Medical Center, Los Angeles, CA; 2 Radiology, Univeristy of California 

Davis Medical Center, Sacramento, CA; 3 University of Florida 

Health Science Center, Jacksonville, FL 

Aim: Sarcopenia is a marker of frailty and predicts mortality 

in cirrhotic patients. Our aim was to determine if sarcopenia 

predicted mortality after transjugular intrahepatic portosystemic 

shunt (TIPS) placement, when adjusting for Model for Endstage 

Liver Disease (MELD) score Methods: We retrospectively 

reviewed patients who underwent TIPS procedure at a single 

center from 2003-2014. CT images were used to determine 

sarcopenia, using measurement of L3 skeletal muscle index 

compared to gender specific thresholds of ≤38.5 cm2/m2 for 

women and ≤52.4 cm2/m2 for men. Cox proportional hazards 

regression was used to determine risk factors for mortality 

after TIPS. Results: A total of 96 patients were included, 

of which 52 had sarcopenia and 44 did not. Median length 

of follow-up was 303 days (range 3-4035 days). Indication 

for TIPS were primarily ascites (n=34, 35.8%) and variceal 

bleeding (n=56, 58.9%). Sarcopenic patients had a greater 

proportion of males (77% vs 38%, p


Myocardial blood flow pre and post-exercise in patients 

with compensated cirrhosis compared with healthy controls 

(HV=healthy, CLD=liver disease) 

6.2, P=0.001, and OR 7.0, 95% CI 2.4-21, P


for primary and secondary prophylaxis should target this predisposition 

and points to an important area of future research. 

Disclosures: 



The following authors have nothing to disclose: Rosaria Spinella, Rohit Sawhney, 

Peter Holland-Fischer, Naina Shah, Rajeshwar Mookerjee 

1483 

Progressive Loss of Hematopoietic Stem Cells and Associated 

Niche Cells in Advanced Cirrhosis 

Chhagan Bihari, Sheetalnath B. Rooge, Dhananjay Kumar, Priyanka 

Saxena, Lovkesh Anand, Smriti Shubham, Archana Rastogi, 

Anupam Kumar, Shiv K. Sarin; Institute of Liver and Biliary Sciences, 

Delhi, India 

Background: Bone marrow (BM) is a reservoir for immunological 

and hematological cells and dysfunction of both has been 

reported in cirrhosis. The mechanisms underlying these dysfunctions 

are not well understood. It is important to know the state 

of BM in cirrhotics, as hepatic regeneration using growth factors 

shown some promise. Aim: To study the effects of disease 

severity and portal hypertension on bone marrow response in 

cirrhosis. Patients and Methods: BM were done as part of a 

work up of cirrhotics (n=168) undergoing regenerative therapy 

protocol or pancytopenia over a period of 4 years. BM without 

any pathology from non-liver, pyrexia of unknown origin cases 

served as controls (n=44). BM assessed for routine examination 

and immunohistochemistry (IHC) for HSCs (CD34), and 

associated niche cells using cell type specific markers; Nestin 

(mesenchymal stem cells), S-100(Schwann cells), GFAP(neural 

fibres). BM HSCs were enumerated by fluorescence activated 

cell sorting. Cytokines and growth factors were analyzed in 

BM plasma (Cirrhosis=26, control= 6). Results: Routine BM in 

cirrhotics showed low cellularity, greater reticulinization, erythroid 

colony disarray as compared to controls. IHC showed 

significant reduction in CD34+ HSCs with increase in MELD (r= 

-0.545; p


1485 

Development of a Novel Clinician Reported Outcome 

Tool for Assessment of Hepatic Encephalopathy 

Jasmohan S. Bajaj 1,2 , Nathan M. Bass 3 , R Todd Frederick 4 , 

Karin Coyne 5 , Mary Kay Margolis 5,6 , Dion F. Coakley 7 , Masoud 

Mokhtarani 7 , Marzena Jurek 7 , Bruce F. Scharschmidt 7 ; 1 Virginia 

Commonwealth University, Richmond, VA; 2 McGuire VA Hospital, 

Richmond, VA; 3 University of California, San Francisco, CA; 

4 California Pacific Medical Center, San Francisco, CA; 5 Evidera, 

Bethesda, MD; 6 Patient-Centered Outcomes Research Institute 

(PCORI), Washington, DC; 7 Horizon Therapeutics (formerly Hyperion 

Therapeutics, Inc.), Brisbane, CA 

The West Haven scale (WH), used for decades in clinical practice, 

was neither developed as an outcome assessment tool 

using methodology acceptable to regulatory authorities, nor 

is it consistently applied. Although recent ISHEN and EASL/ 

AASLD Guidelines have clarified HE nomenclature and treatment, 

there is no standardized tool for assessing overt HE 

(OHE). A standardized clinician reported outcome (ClinRO) 

tool to diagnose and grade OHE was developed, in compliance 

with methodologies accepted by regulatory authorities, 

for use in clinical trials as well as daily clinical practice. Study 

Design: A two part iterative Delphi method was employed 

using a 3-member steering committee and a panel of 40 

practicing hepatologists in the US, Canada, Europe, South 

America, and Australia to develop a new ClinRO instrument 

(HE Grading Instrument, HEGI) for OHE identification and 

grading. Part 1 involved open-ended concept elicitation from 

the panel regarding clinical findings they use in their daily 

practice to diagnose and rate OHE severity. Part 2 included 

testing the HEGI for clarity, usability, and inter- and intra-rater 

reproducibility. To assess inter-rater reproducibility, 34 panelists 

viewed 5 videos depicting HE episodes of differing severity 

and determined the presence/absence of OHE, as well as its 

severity. Intra-rater reproducibility was assessed among a subset 

of the panelists who viewed the video vignettes a second 

time, in random order, and rated each episode again. Results: 

Of the 40 panelists, 97.5% had participated in clinical trials, 

52.5% saw > 10 HE patients/week, and 32% were from outside 

the US. Major OHE criteria recommended in Part 1 for 

inclusion in the HEGI included disorientation to time (90%), 

place (72.5%), and person (70.0%), asterixis (97.5%), lethargy 

(92.5%), and coma (100%). Assuming exclusion of other 

causes, consensus was reached that any of the following constituted 

a minimum requirement for diagnosis of OHE: (1) any 

disorientation, (2) presence of both lethargy and asterixis, or 

(3) coma. The overall adjudicated concordance in Part 2 for 

discriminating between presence/absence of OHE using HEGI 

was 97.1%; the overall concordance for rating episode severity 

was 87.1%; and the overall concordance for reproducibility 

was 98%. Conclusions: The HEGI, a ClinRO tool for diagnosis 

and grading of OHE episodes, was developed using the principles 

contained in the FDA Guidance for Development of Patient 

Reported Outcomes for instrument validity and reproducibility. 

It is anticipated to aid in the identification of OHE in clinical 

practice and enhance the reliability of endpoint assessment in 

clinical trials for OHE. 

Disclosures: 




Nathan M. Bass - Advisory Committees or Review Panels: Quest Diagnostics 



Karin Coyne - Consulting: Evidera 




The following authors have nothing to disclose: Mary Kay Margolis, Bruce F. 

Scharschmidt 

1486 

Usefulness of Balloon-Occluded Retrograde Obliteration 

(B-RTO) as a Consolidation Procedure after Anticoagulation 

Therapy in Cirrhotic Patients with Portal Vein 

Thrombosis 

Mie Inao, Kazuki Hirahara, Kayoko Sugawara, Nobuaki 

Nakayama, Yukinori Imai, Satoshi Mochida; Department of Gastroenterology 

& Hepatology, Saitama Medical University, Moroyama-Machi, 

Japan 

Aim: Although anticoagulation therapies with Xa inhibitors and 

antithrombin concentrates were shown to be effective for attenuation 

of portal vein thrombosis in cirrhotic patients, aggravation 

or recurrence of the lesions may occur following the 

therapies leading to derangement of liver function. Decrease 

of blood flow in the portal vein as a consequence of porto-systemic 

shunts may responsible for thrombosis development. 

Thus, the usefulness of B-RTO as a consolidation procedure 

after anticoagulation therapies was evaluated. Methods: The 

subjects were 43 patients (23 men and 20 women, aged from 

40 to 76 years old) with liver cirrhosis complicating portal vein 

thrombosis. Both danaparoid Na (2,500 units/day) and antithrombin 

concentrates (1,500 units/day) were intravenously 

administrated for 3 days followed by danaparoid Na injections 

for further 11 days. Patients seen in April 2013 and later 

received B-RTO procedures after anticoagulation therapies, 

when porto-systemic shunts were observed on CT and/or MRI 

imaging. A balloon catheter was inserted into the shunts followed 

by injection of 5% ethanolamine oleate through the catheter 

under balloon inflation. The balloon was kept inflation for 

6 to 48 hours depending on sizes of the shunts. Results: Immediately 

after anticoagulation therapies, portal vein thrombosis 

was completely disappeared in 11 patients (25%) and the 

sizes of thrombosis were attenuated in 15 patients (35%), while 

the lesions did not change in 17 patients (40%). B-RTO was 

additionally done in 4 patients; 2 patients showing complete 

thrombosis disappearance and 2 patients failing to achieve 

thrombosis attenuation. Following B-RTO procedures, thrombosis 

did not recur in both of the former patients and the lesions 

disappeared in both of the latter patients despite that anticoagulation 

therapies were ineffective. In contrast, in 39 patients 

without additional B-RTO procedures, thrombosis recurred in 

4 among 9 patients after thrombosis disappearance and was 

aggravated in 6 among 15 patients achieving thrombosis 

attenuation. Conclusion: B-RTO was effective as a consolidation 

procedure after anticoagulation therapies for patients with 

portal vein thrombosis even in those failing to achieve attenuation 

of the lesions when porto-systemic shunts responsible for 

decrease of blood flows in the portal vein were observed. 

Disclosures: 



The following authors have nothing to disclose: Mie Inao, Kazuki Hirahara, 

Kayoko Sugawara, Nobuaki Nakayama, Yukinori Imai


1487 

Osteopontin is a new prognostic marker of liver cirrhosis 

Radan Bruha 1 , Marie Jachymova 2 , Jaromir Petrtyl 1 , Karel Dvorak 1 , 

Martin Lenicek 2 , Petr Urbanek 3 , Libor Vitek 2 ; 1 4th Intenal Clinic, 

Charles University in Prague, 1st Faculty of Medicine, Prague, 

Czech Republic; 2 Institute of Clinical Biochemistry and Laboratory 

Diagnostics, Charles University in Prague, 1st Faculty of medicine, 

Prague, Czech Republic; 3 Internal Clinic, Central Military Hospital, 

Prague, Czech Republic 

Background and Aims: Portal hypertension leads to major 

complications of cirrhosis. Invasively measured hepatic venous 

pressure gradient (HVPG) is a strong prognostic indicator in 

patients with cirrhosis and portal hypertension. Recently, osteopontin 

has emerged as a new marker with a possible relation 

to fibrosis and cirrhosis and close correlation to portal hypertension. 

Our aim was to evaluate the relationship of osteopontin 

plasma concentrations to the prognosis of patients with 

cirrhosis. Methods: A cohort of 154 patients with liver cirrhosis 

(112 ethylic, 108 men, age 34-72 years) were enrolled. The 

HVPG was measured by standard catheterization using the 

balloon wedge technique. The mean follow-up of the patients 

was 3.7±2.6 years (maximal length 7 years). Osteopontin was 

measured by ELISA method. Results: The mean value of HVPG 

was 16.18±5.6 mmHg. Patients with osteopontin values above 

80 ng/ml had a significantly lower cumulative survival rate 

compared to those with osteopontin ≤80 ng/ml (37% vs. 56%, 

p=0.00035; OR 2.23, 95% CI 1.06-4.68; Fig 1). Similarly, 

the HVPG cut-off value of 10 mm Hg divided patients into 2 

groups with significantly different probabilities of cumulative 

survival (39% for those with HVPG>10 mm Hg compared to 

65% for those with HVPG≤10 mm Hg; p=0.0086, OR 2.92, 

95% CI 1.09-7.76). Mortality in patients with at least one risk 

factor (HVPG above 10 mm Hg or plasma OPN above 80 

ng/l) was more than twice as high compared to patients without 

any risk factors (OR=2.34); in those with both risk factors, 

mortality was more than five times as high (OR=5.1) compared 

to patients without any risk factors. Conclusions: Osteopontin is 

a strong prognostic factor for overall survival in patients with 

cirrhosis. Supported by SVV 260 156/2015. 

Figure 1: Cumulative proportion of surviving patients with plasma 

OPN levels below and above 80 ng/ml using the Kaplan-Meier 

method. 

1488 

Liver Injury Independently Predicts Incident Cardiovascular 

Disease (CVD) Risk 

Kaku So-Armah 1 , Janet Tate 3,2 , Joseph K. Lim 2 , Vincent Lo Re 14 , 

Vincent Marconi 4 , Jeffrey Samet 1 , Cynthia Gibert 8 , David Leaf 9,10 , 

Adeel Butt 6,7 , Matthew B. Goetz 9,10 , David Rimland 4,11 , Maria C. 

Rodriguez-Barradas 12,13 , Matthew Budoff 5 , Chung-Chou Chang 15 , 

Amy Justice 2,3 , Matthew Freiberg 16 ; 1 Boston University School of 

Medicine, Boston, MA; 2 Yale University School of Medicine, New 

Haven, CT; 3 VA Connecticut Healthcare System, West Haven, 

CT; 4 Emory University School of Medicine, Atlanta, GA; 5 LA 

Biomed Research Institute, Torrance, CA; 6 VA Pittsburgh Healthcare 

System, Pittsburgh, PA; 7 Hamad Healthcare Quality Institute 

and Hamad Medical Corporation, Doha, Qatar; 8 Washington DC 

VA Medical Center, Washington, DC; 9 David Geffen School of 

Medicine at UCLA, Los Angeles, CA; 10 VA Greater Los Angeles 

Healthcare System, Los Angeles, CA; 11 Atlanta VA Medical Center, 

Atlanta, GA; 12 Michael E. DeBakey VA Medical Center, Houston, 

TX; 13 Baylor College of Medicine, Houston, TX; 14 University of 

Pennsylvania School of Medicine, Philadelphia, PA; 15 University of 

Pittsburgh Medical Center, Pittsburgh, PA; 16 Vanderbilt University 

School of Medicine, Nashville, TN 

Background: Multiple etiologies of liver injury are associated 

with cardiovascular disease (CVD) e.g., hepatitis C or B (HCV, 

HBV) and HIV co-infection, alcohol, and obesity. We hypothesize 

that liver injury is a mechanism of increased CVD risk. 

We assessed whether liver fibrosis index 4 (FIB4) predicted 

incident CVD. Liver-related comorbidities may have extra-hepatic 

contribution to CVD risk. Thus, we performed sensitivity 

analyses excluding people with HCV, HBV, HIV, alcohol use 

disorder, and obesity. Methods: Participants from the Veterans 

Aging Cohort Study Virtual Cohort (VACS VC), without CVD 

at baseline (first clinical visit after 4/1/2003) were included. 

Liver injury was categorized using baseline FIB4 (calculated 

using age, liver transaminases and platelets) and ICD-9 codes 

for cirrhosis and hepatic decompensation. Total non-fatal and 

fatal CVD was assessed using VA, VA Fee For Service and 

Medicare ICD-9 and procedure codes for myocardial infarction, 

heart failure, coronary heart disease and stroke, and 

National Death Index cause of death data. Follow-up ended 

after a CVD event, or death, or on 9/30/2012. Cox regression 

analyses were used to estimate CVD risk adjusting for 

confounders including renal disease. Results: After excluding 

21488 people with prevalent CVD, 104731 people remained. 

Mean (SD) age was 49.6 (9.5) years in this largely male (90%) 

cohort followed for a median (IQR) of 7 (3, 9) years. LDL cholesterol 

and body mass index decreased with increasing FIB4. 

African American race, HIV, HCV, HBV, current smoking, type 

2 diabetes, alcohol use disorder, cocaine use, and anemia 

were more common among those with FIB4>1.45 than those 

with FIB43.25 or a clinical diagnosis of cirrhosis or 

hepatic decompensation had significantly elevated CVD risk 

despite also having higher mortality rates (see Table below). 

This association persisted among those without heart failure or 

without HCV, HBV, HIV, alcohol abuse/dependence diagnosis 

and BMI>30 kg/m 2 . Conclusions: FIB4 and severe liver injury 

independently predict risk of CVD in VACS VC. Future studies 

should assess if treatment of liver injury reduces CVD risk. 

Disclosures: 

The following authors have nothing to disclose: Radan Bruha, Marie Jachymova, 

Jaromir Petrtyl, Karel Dvorak, Martin Lenicek, Petr Urbanek, Libor Vitek


GPC in culture-positive SBP and/or bacteremia were observed. 

Patients with GNB had more advanced liver disease and more 

severe inflammatory response compared to GPC ones. A variety 

of drug-resistant microorganisms and a high rate of E. Faecium 

have emerged. Due to high rates of resistance in currently 

recommended therapy and prophylaxis, the choice of optimal 

antibiotic therapy should be individualized. 

Disclosures: 

The following authors have nothing to disclose: Alexandra Alexopoulou, Larisa E. 

Vasilieva, Sotiria Papadaki, Danai Agiasotelli, Sophia Pouriki, Athanasia Tsiriga, 

Marina G. Toutouza, Spyros P. Dourakis 

Disclosures: 




Matthew Budoff - Grant/Research Support: GE, NIH 

The following authors have nothing to disclose: Kaku So-Armah, Janet Tate, Vincent 

Lo Re, Vincent Marconi, Jeffrey Samet, Cynthia Gibert, David Leaf, Adeel 

Butt, Matthew B. Goetz, David Rimland, Maria C. Rodriguez-Barradas, Chung- 

Chou Chang, Amy Justice, Matthew Freiberg 

1489 

Increasing frequency of gram-negative multi-drug resistant 

bacteria and Enterococcus faecium in spontaneous 

bacterial peritonitis and bacteremia in patients with 

decompensated cirrhosis 

Alexandra Alexopoulou 1 , Larisa E. Vasilieva 1 , Sotiria Papadaki 1 , 

Danai Agiasotelli 1 , Sophia Pouriki 1 , Athanasia Tsiriga 2 , Marina 

G. Toutouza 2 , Spyros P. Dourakis 1 ; 1 2nd Dept of Medicine, Medical 

School University of Athens, Athens, Greece; 2 Microbiology 

Department Hippokration GNA, Athens, Greece 

Purpose/Background: Spontaneous bacterial peritonitis (SBP) 

and bacteremia in cirrhotic patients are historically caused 

by Gram-negative bacteria (GNB) almost exclusively Enterobacteriaceae. 

Recently, an increasing rate of infections with 

Gram-positive cocci (GPC) and multi-drug resistant (MDR) 

microorganisms was demonstrated. The purpose of this study 

was to assess possible recent changes of the bacteria causing 

SBP or bacteremia in patients with decompensated cirrhosis. 

Methods: We prospectively recorded 130 cases (68.5% 

males) with culture-positive SBP or spontaneous bacteremia 

without SBP during a 3-year-period (2012-2014). Results: The 

76.2% of patients had health care-associated (HCA) or nosocomial 

infections. GPC were found in half of the cases. The most 

prevalent organisms in a descending order were E. coli (33), 

Enterococcus spp (30, including 17 E. Faecium), Streptococcus 

spp (25), K. pneumonia (16), S. aureus (8), P. aeruginosa 

(5), other GNB (11) and anaerobes (2). Twenty five (19.2%) 

of the isolated bacteria were MDR, including extended-spectrum-beta-lactamase-producing 

(ESBL)-GNB (9), carbapenemase-producing 

(KPC)-K. pneumonia (5), P. aeruginosa (5), A. 

baumannii (2), E. Faecium Vancomycin-resistant (2), KPC colistin-Resistant-K. 

pneumonia (1) and metallo-Β-Lactamase producing 

(MBL)-E. Coli (1). MDR bacteria were more frequently 

isolated in HCA/nosocomial than in community-acquired infections 

(100% vs 70.5%) (P=0.002). E. Faecium was always 

associated with HCA/nosocomial infections. Patients with MDR 

bacteria or E. Faecium had longer hospitalization compared 

to the rest of patients (P=0.013). Patients with GNB had more 

advanced liver disease and higher neutrophil-to-leucocyte ratio 

compared to those with GPC [MELD 21 (17-26) vs 18 (13- 

25) respectively, (P=0.059), total bilirubin 4.98 (2,2-11,1) vs 

2.75 (1.79-5.90) respectively, (P=0.046] and [84 (74-91) vs 

78 (69-85) respectively, (P=0.018)]. Third-generation cephalosporin-resistance 

was observed in 43.1% and quinolone-resistance 

in 49.6%. Conclusion: Similar frequencies of GNB and 

1490 

Non-invasive assessment of portal hypertensive gastropathy: 

feasibility of contrast-enhanced ultrasonography 

for stomach wall 

Hitoshi Maruyama, Soichiro Kiyono, Takayuki Kondo, Tadashi 

Sekimoto, Osamu Yokosuka; Department of Gastroenterology 

and Nephrology, Chiba University Graduate School of Medicine, 

Chiba, Japan 

Background/Aim: Portal hypertensive gastropathy (PHG) is one 

of the major complications in cirrhosis, and endoscopy may be 

the only diagnostic tool. The analysis of microbubble-induced 

enhancement of the stomach wall on the sonogram may detect 

the hemodynamic difference between patients with and without 

PHG. The aim was to elucidate the efficacy of transabdominal 

contrast-enhanced ultrasound (CEUS) as a non-invasive 

diagnostic tool of PHG. Methods: This is a prospective study 

performed in 54 subjects, 40 cirrhosis patients (60.4 ± 10.5 

years; 11 females; Child A 25, B 15) with esophageal varices 

who underwent the measurement of hepatic venous pressure 

gradient (HVPG) and 14 controls (41.4 ± 13.1 years; female 

4). Contrast effects using perflubutane microbubble agent 

were assessed at the upper stomach wall by CEUS. The peak 

enhancement time (PT, time from contrast onset to maximum 

enhancement) and the intensity ratio between pre-enhancement 

and peak-enhancement (IR) were calculated using time-intensity 

curve. Results: PHG was detected in 15 cirrhosis patients 

(37.5 %; mild 12, severe 3). The PT and IR were 8.5s and 

8.3s (p=0.348), and 1.22 and 1.56 (p


Disclosures: 

The following authors have nothing to disclose: Hitoshi Maruyama, Soichiro 

Kiyono, Takayuki Kondo, Tadashi Sekimoto, Osamu Yokosuka 

1491 

Videogame Training Improves Brain Plasticity In Covert 

Hepatic Encephalopathy Through Changes In Brain Connectivity 

Vishwadeep Ahluwalia, James Wade, Frederick G. Moeller, Joel 

L. Steinberg, Michael Lennon, Ariel Unser, Melanie White, Douglas 

M. Heuman, Edith A. Gavis, Jasmohan S. Bajaj; VCU and 

McGuire VAMC, Richmond, VA 

Cognitive improvement using videogames could be a non-pharmacologic 

covert HE(CHE) therapy but its effect on brain networks 

is unclear. Aim: Assess the impact of videogame-related 

cognitive therapy on brain networks in CHE. Methods: CHE 

cirrhotics underwent an 8-wk trial with 4 visits with CHE-related 

[Paper/pencil & inhibitory control test (ICT)] & unrelated 

tests (Hopkins Verbal learning test HVLT). Visit 1 & 2 were 2 

wks apart to gauge learning.At visit 2 an iPod loaded with 2 

games; IQ Boost & Arcade Challenge was given for 4 wks. Visit 

3 was 4 wks post-iPod when cognitive testing was repeated & 

trial ended. A subgroup (n=13) underwent MRI at visit 2 & 3 

to define functional/structural connectivity change. Changes in 

brain MRI & tests related/unrelated to videogames were analyzed.Resting 

state analysis produces functional brain network 

maps linked with attention, memory &other domains. Tractography 

estimates the spatial probability distribution of white 

matter tracts between two regions;it was performed between 

memory-related (HVLT-related) regions such as hippocampus, 

angular (AG) & anterior middle temporal gyri (AMTG). Pre-/ 

post-ipod differences between networks were tested after 

region of interest creation. Results: 20 CHE pts (12 men, 56 

yrs, MELD 10) were recruited. There was a significant improvement 

in paper-pencil tests between visits 1 & 2. ICT & HVLT 

improved only during the iPod phase. 27% of pts’ performance 

normalized at visit 4 (p


Disclosures: 

The following authors have nothing to disclose: Tsuyoshi Ishikawa, Ryo Sasaki, 

Tatsuro Nishimura, Yuki Aibe, Shogo Shiratsuki, Kazuhito Matsunaga, Takuya 

Iwamoto, Taro Takami, Isao Sakaida 

1493 

Inadequate Food And Water Intake In Patients With Cirrhosis: 

Need Of A Personalized Nutritional Approach 

Ferdinando A. Giannone 1,3 , Marco Domenicali 1,3 , Maurizio Baldassarre 

1,3 , Paola De Pasquale 1 , Silvia Boffelli 1,2 , Maristella Laggetta 

1,3 , Maurizio Biselli 1,2 , Mauro Bernardi 1,2 , Paolo Caraceni 1,3 ; 

1 Department of Surgical and Medical Sciences, Alma Mater Studiorum 

University of Bologna, Bologna, Italy; 2 U.O. Semeiotica 

Medica, Azienda Ospedaliero-Universitaria di Bologna - Policlinico 

S. Orsola-Malpighi, Bologna, Italy; 3 Center for Applied 

Biomedical Research (CRBA), Alma Mater Studiorum University of 

Bologna, Bologna, Italy 

Background/Aim. Malnutrition is a frequent complication in 

patients with cirrhosis, contributing to excess morbidity and 

mortality. Although practical guidelines have been published, 

its management at the individual level is quite complex since 

cirrhotic patients constitute a very heterogeneous population in 

terms of needs, intake and utilization of nutrients as well as motivation 

and adherence to dietary programs. This study aimed to 

assess whether the food and water intake of cirrhotic patients 

is adequate to the needs of their specific clinical condition 

according to the recommended nutritional indications. Methods. 

Outpatients with cirrhosis were consecutively screened 

for inadequate nutrition by using a scored algorithm, including 

as risk factors altered BMI, recent body weight changes, 

presence of diabetes, ascites or hepatic encephalopathy (HE). 

All patients at risk were offered to be evaluated by a certified 

nutritionist. Patients were then asked to fill up a weekly food 

diary, which provided information on the amount and origin 

of the food, water intake, and, physical activity, and to perform 

a hand-grip strength (HGS) test to assess muscle wasting. 

Results. Of the 144 pts screened, only 16% showed no risk of 

inadequate nutrition, while 37% presented 1 risk factor and 

47% ≥2 risk factors. Child-Pugh and MELD scores were 5.5 

and 7.4 in the group with no risk, 6.8 and 11.7 in pts with 

one risk factor, and 7.6 and 12.8 in those with ≥2 risk factors. 

Of the 121 pts at risk, 86 pts accepted the nutritional evaluation: 

reduced muscle strength was found in 56% of pts, while 

65% properly returned the weekly food diary. Interestingly, 

the adherence was higher in those with ≥2 risk factors and/or 

more advanced disease, particularly if transplant candidates. 

Despite all patients had received general nutritional indications 

by their hepatologists, the analysis of the diary showed that 

70% of pts had an inadequate intake of calories, 41% of water 

and 36% of salt. Daily and/or weekly distribution of macronutrients 

was also inadequate in the vast majority: carbohydrates 

in 88%, sugar in 73%, proteins in 86%, lipids in 36 %, and 

fibers in 95%. No physical activity was present in 79% of pts. 

Conclusions. This study shows that the vast majority of cirrhotic 

patients have an inadequate intake of food, water and macronutrients, 

which was often associated to muscle wasting and 

often related to a scarce knowledge of nutritional facts. This 

highlights the need of a personalized nutritional approach, 

including educational and motivational aspects, which takes 

into account the complex interacting, often conflicting, nutritional 

needs of each individual patient. 

Disclosures: 

Paolo Caraceni - Advisory Committees or Review Panels: GSK; Consulting: BMS; 

Grant/Research Support: Grifols; Speaking and Teaching: Baxter, Kedrion 

The following authors have nothing to disclose: Ferdinando A. Giannone, Marco 

Domenicali, Maurizio Baldassarre, Paola De Pasquale, Silvia Boffelli, Maristella 

Laggetta, Maurizio Biselli, Mauro Bernardi 

1494 

Myocardial strain as a noninvasive measure of circulatory 

dysfunction and systolic function in liver cirrhosis. 

Carolina F. Pimentel 1,4 , Maria Lucia Ferraz 1 , Miguel Osman D. 

Aguiar 2 , Adriano M. Gonzalez 1,2 , Gabriel D. Branco 2 , Marcel 

Superbia 2 , Ana Cristina A. Feldner 1 , Michelle Lai 5 , Wilson 

Mathias 3 , Mario Kondo 1 ; 1 Gastroenterology, Federal University of 

Sao Paulo, Sao Paulo, Brazil; 2 Echocardiology, Sao Luiz Hospital, 

Sao Paulo, Brazil; 3 Heart Institute, University of Sao Paulo, Sao 

Paulo, Brazil; 4 Liver Transplant, Euryclides de J Zerbini Transplant 

Hospital, Sao Paulo, Brazil; 5 Gastroenterology and Hepatology, 

Beth Israel Deaconess Medical Center, Harvard Medical School, 

Boston, MA 

Cirrhosis is related to hyperdynamic circulation with insufficient 

organ perfusion, activation of RAAS and compensatory 

increase in cardiac output. Left ventricular longitudinal myocardial 

strain is a non-invasive echocardiographic marker of 

myocardial contractility, able to identify subclinical systolic dysfunction 

(SD). We evaluated cirrhotic patients and measured 

strain rate, parameters for cardiac dysfunction and serum renin 

levels and analyzed them by Child Classes. Method: A cohort 

of 103 cirrhotic outpatients was studied. SD was defined as 

strain>-18% or ejection fraction (EF)< 55%. and diastolic dysfunction 

(DD) findings as E/A8. Results: Non-alcoholic 

liver cirrhosis was the most common etiology (70%). 

Child stages were A (66%), B (21%) and C(13%). Alcohol 

consumption was unrelated to any parameters. SD was more 

frequently diagnosed by strain rate than EF (5% vs. 0.9%). 

All patients had normal ventricular measurements and 24% 

evidence of DD. These patients had strain rates consistent with 

decreased myocardial contractility, even without anatomical 

abnormalities. Both strain rate and serum renin levels were 

significantly correlated with Child scores (p=0.002, r=-0.3; 

p=0.03, r=0.3, respectively), showing increased myocardial 

contractility and progressive RAAS activation with increasing 

Child scores. Mean strain rates and mean serum renin levels 

were statistically different among Child classes (see figure). 

Conclusions: Myocardial strain rate and serum renin level correlated 

with Child stages consistent with a progressively more 

hyperdynamic circulatory state with worsening liver decompensation. 

Strain rate was more sensitive than ejection fraction in 

diagnosing SD and is a promising tool for early diagnosis of 

subclinical systolic dysfunction and circulatory dysfunction in 

cirrhosis.


this cohort, with 64% of the cohort having at least one highrisk 

parameter. Table 1 shows prevalence of high risk HRV 

parameters. SDNN and SDANN correlated with Child scores 

(p


and dry BMI, mid-arm muscle circumference, SMI and VMI. 

Logistic regression analysis revealed that INR [O.R. 6.581 

(95% C.I. 1.330 – 32.569), p = 0.021] and log-transformed 

leptin [O.R. 0.425 (0.221 – 0.816), p < 0.010] were independently 

associated with malnutrition. Conclusions: Low serum 

leptin and elevated INR are associated with malnutrition in hospitalized 

patients with cirrhosis and may represent biomarkers 

for early identification of malnutrition in this population. 

Disclosures: 

The following authors have nothing to disclose: Vikrant Rachakonda, Jaideep 

Behari 

Disclosures: 




The following authors have nothing to disclose: Thoetchai Peeraphatdit, Niyada 

Naksuk, Roongruedee Chaiteerakij 

1497 

The World Congress of Gastroenterology Guidelines for 

the Diagnosis of Left Ventricular Diastolic Dysfunction 

Differ from the American Society of Echocardiography 

Standard Recommendation 

Thoetchai Peeraphatdit 1,2 , Niyada Naksuk 3 , Lewis R. Roberts 2 , 

Roongruedee Chaiteerakij 2,4 ; 1 Internal Medicine, University of 

Minnesota, Minneapolis, MN; 2 Gastroenterology and Hepatology, 

Mayo Clinic College of Medicine, Rochester, MN; 3 Cardiovascular 

Diseases, Mayo Clinic College of Medicine, Rochester, 

MN; 4 Medicine, Chulalongkorn University, Bangkok, Thailand 

Purpose: Diagnostic criteria for left ventricular diastolic dysfunction 

(LVDD) proposed by the World Congress of Gastroenterology 

(WCG) has never been tested and are different from 

the American Society of Echocardiography (ASE) criteria. This 

study determined the agreement of LVDD diagnosed by these 

two guidelines, the performance of the WCG criteria, and 

their prognostic values. Methods: We retrospectively enrolled 

914 consecutive patients with normal left ventricular ejection 

fraction (LVEF) and available data on diastolic function undergoing 

liver transplant (LT) at a tertiary medical center between 

1994 and 2014. According to the ASE, LVDD was diagnosed 

based on a stepwise approach using multiple parameters. 

However, by the WCG, LVDD was defined as an early to 

late ventricular filling velocities ratio (E/A) 200 ms. The agreement of LVDD diagnosed by 

the two guidelines was analyzed using the Kappa statistic. 

The performance of the WCG criteria was determined using 

the ASE criteria as a gold standard. Results: Mean age was 

53.7±10.6 years; 65.2% were male. During 5.6±4.2 years 

follow up after LT, 21.6% patients had died. Overall LVEF was 

65.8±5.1%. By the ASE criteria, 47.2% of patients had LVDD; 

whereas the prevalence of LVDD was 19.6%, 61.0%, 64.5% 

and 13.9% by the WCG criteria of E/A 200 ms, 

either one of the criteria, or both criteria, respectively. There 

was minimal agreement between LVDD diagnosed by the 2 

WCG criteria (Kappa=0.10). Moreover, there was generally 

poor or slight agreement between the ASE and the WCG criteria 

(Kappa=0.08-0.18, Table). The WCG criteria had sensitivities 

of 50-70% and specificities of 58-62% (Table). However, 

in the multivariate Cox regression, only LVDD diagnosed by 

the WCG criterion of E/A


in sickle hepatopathy. Despite diagnostic challenges with HB 

in SCD, DB, ALP and PLT appear to be respectively associated 

with increasing portal pressures and mortality. 

Disclosures: 

The following authors have nothing to disclose: David M. Chascsa, Ohad Etzion, 

Hawwa Alao, Courtney Fitzhugh, John F. Tisdale, Caterina Minniti, Matthew 

Hsieh, David E. Kleiner, Ahmed M. Gharib, Varun K. Takyar, Jason L. Eccleston, 

Theo Heller, Christopher Koh 

1499 

A half of hepatic hydrothorax in cirrhotic patients were 

caused by porous diaphragm syndrome due to radiofrequency 

ablation for hepatocellular carcinoma 

Kenichi Miyoshi, Masahiko Koda, Toshiaki Okamoto, Tomomitsu 

Matono, Takaaki Sugihara, Keiko Hosho, Jun-ichi Okano; Tottori 

University School of Medicine, Yonago, Japan 

Aims Hepatic hydrothorax is defined as a significant pleural 

effusion in cirrhotic patients without underlying pulmonary or 

cardiac disease. One of causes of refractory hepatic hydrothorax 

is porous diaphragm syndrome. However, the frequency 

of porous diaphragm syndrome in hepatic hydrothorax and 

the cause of porous diaphragm syndrome are unknown. We 

aimed to evaluate its frequency, clinical background, and therapeutic 

value of them. Methods From August 2005 to April 

2015, 124 cirrhotic patients hospitalized due to hepatic 

ascites are enrolled. Patient’s data hospitalized several times 

during its period were referred to the most serious cases. We 

compared clinical background, and therapeutic value between 

patients with and without hepatic hydrothorax. Four patients 

with the rupture of hepatocellular carcinoma (HCC), 3 hemodialysis 

patients, and 4 patients taking warfarin were excluded. 

In 17 of 115 patients from September 2007 with hydrothorax, 

we examined the presence of diaphragmatic defects with 

intraperitoneal injection of perflubutane (approved by the local 

ethics committee). We analyzed clinical background between 

patients with and without porous diaphragm syndrome. Results 

Forty-one patients (34.2%) in 124 cirrhotic patients with ascites 

had hepatic hydrothorax. There was no difference between 

patients with and without hydrothorax on clinical background 

and therapeutic value. Ten (58.8%) of 17 patients with hydrothorax 

had porous diaphragm syndrome. Accordingly, 10 

(8.7%) of 115 cirrhotic patients with ascites complicated diaphragmatic 

defects. Patients with porous diaphragm syndrome 

had higher serum albumin levels and fewer hepatic encephalopathy 

events, and more past history of radiofrequency ablation 

(RFA) for HCC adjacent to diaphragm than those without 

porous diaphragm syndrome. Multivariate analysis showed 

that only RFA was an independent factor for porous diaphragm 

syndrome. Tour of 10 patients with porous diaphragm syndrome 

underwent thoracoscopic diaphragm plication. In all 

these patients, pleural effusion disappeared or decreased. 

Conclusions 58.8% of patients with hepatic hydrothorax were 

caused by porous diaphragm syndrome. Porous diaphragm 

syndrome was related to RFA for HCC. 

Disclosures: 

The following authors have nothing to disclose: Kenichi Miyoshi, Masahiko 

Koda, Toshiaki Okamoto, Tomomitsu Matono, Takaaki Sugihara, Keiko Hosho, 

Jun-ichi Okano 

1500 

Sarcopenic obesity impairs prognosis of patients with 

cirrhosis 

Motoh Iwasa, Ryosuke Sugimoto, Yoshinao Kobayashi, Yoshiyuki 

Takei; Department of Gastroenterology and Hepatology, Mie University 

Graduate School of Medicine, Tsu, Japan 

Aims: It has been reported that there is a high incidence of serious 

events and poor outcomes when sarcopenia accompanies 

cirrhosis. In terms of body fat, it was recently reported that the 

prognosis is poor after resection of hepatocellular carcinoma 

in cirrhosis patients with decreased visceral fat. In the present 

study, skeletal muscle mass and visceral fat mass of patients 

with cirrhosis were measured, and the relationships between 

these body composition measurements and prognosis were 

investigated. Furthermore, time-dependent changes in body 

composition measurements were also examined. Methods: A 

total of 161 patients with cirrhosis (94 men, 67 women; 67 ± 

9 years) who received medical care and nutritional assessment 

were investigated. For body composition, in addition to height 

and weight, visceral fat area (VFA, cm 2 ) and upper limb skeletal 

muscle mass (kg) were measured using a multi-frequency 

bioelectrical impedance analysis device. Subsequently, setting 

the cut-off value of VFA at 100 cm 2 for visceral obesity and the 

cut-off value of sarcopenia at 1.7 kg/m 2 for men and 1.2 kg/ 

m 2 for women, patients were classified, and their outcomes 

were observed. Furthermore, 60 patients with cirrhosis who 

underwent multiple body composition measurements during 

the observation period were extracted, and their initial and 

final measurements were compared. Results: Although there 

was a weak positive correlation between VFA and albumin, 

significant correlations were not observed between muscle 

mass and blood test results. Patients were classified into the 

following 4 groups: normal body composition; visceral obesity; 

sarcopenia; and sarcopenic obesity. There were 60, 61, 25, 

and 15 patients, respectively, in each group. Sarcopenia and 

sarcopenic obesity were more common in men, and sarcopenic 

obesity was more common in elderly individuals. In the comparison 

of 3 groups (visceral obesity, sarcopenia, and sarcopenic 

obesity groups), 22 (36%), 12 (48%), and 10 (67%) deaths 

were noted, respectively, during a mean observation period 

of 1005 days, and the prognosis was significantly worse in 

sarcopenic obesity, followed by sarcopenia and visceral obesity, 

in that order (P < 0.05). A significant positive correlation 

was observed only between percent change in skeletal muscle 

mass and percent change in serum albumin. Sarcopenia is a 

common feature of advanced cirrhosis, and transitions were 

observed from normal body composition to sarcopenia and 

from obese to sarcopenic obesity. Conclusions: Body composition 

is a prognostic factor for cirrhosis, and a better body 

composition may be advantageous for long-term survival in 

patients with cirrhosis. 

Disclosures: 

The following authors have nothing to disclose: Motoh Iwasa, Ryosuke Sugimoto, 

Yoshinao Kobayashi, Yoshiyuki Takei


1501 

Portal vein thrombosis, mortality and hepatic decompensation 

in patients with cirrhosis: A meta-analysis 

Jonathan G. Stine 1 , Puja M. Shah 2 , Scott L. Cornella 3 , Sean R. 

Rudnick 1 , George R. Stukenborg 4 , Patrick Northup 1 ; 1 Gastroenterology 

& Hepatology, University of Virginia, Charlottesville, VA; 

2 Surgery, University of Virginia, Charlottesville, VA; 3 Medicine, 

University of Virginia, Charlottesville, VA; 4 Public Health Sciences, 

University of Virginia, Charlottesville, VA 

Background: Non-neoplastic portal vein thrombosis is a common 

complication of cirrhosis. Treatment options can be risky 

and are not without complications. To make the most appropriate 

choice, clinicians need to know what the evidence 

says about the clinical importance of portal vein thrombosis 

in patients with cirrhosis. Objectives: To determine the clinical 

impact of portal vein thrombosis in terms of both mortality 

and hepatic decompensation (variceal hemorrhage, ascites, 

portosystemic encephalopathy) in adult patients with cirrhosis. 

Methods: We identified observational research studies 

through February 2015 from MEDLINE, EMBASE, Scopus, Science 

Citation Index, AMED, Google Scholar, the Cochrane 

Library and the relevant grey literature. Two independent 

reviewers screened citations and extracted data. We specifically 

excluded any studies involving liver transplantation. We 

graded the strength of evidence and determined the magnitude 

of effect by calculating DerSimonian and Laird random effects 

odds ratios to obtain aggregate estimates of effect size and 

95% confidence intervals. Between-study variability and heterogeneity 

were assessed. Main results: After reviewing 226 

citations, we included 3 studies with 2,436 participants. The 

majority of included patients were Caucasian. Viral hepatitis 

was the most common etiology of cirrhosis (42%) and alcoholic 

liver disease was the second most prevalent (21%). Median 

MELD scores ranged from 10-15 for each individual study. The 

prevalence of portal vein thrombosis in aggregate was 7.2%. 

Patients with portal vein thrombosis had an increased risk of 

mortality (OR 1.69, 95% CI 1.18-2.42, p


1503 

Real Time Pressure Volume Loops in Cirrhosis: Characterization 

of Systolic and Diastolic Function and Validation 

of Doppler Indices with the Gold Standard 

Cristina Ripoll 2,1 , Raquel Yotti 3 , Yolanda Benito 3 , Diego Rincón 2 , 

Maria Vega Catalina 2 , Jaime Elizaga 3 , Javier Bermejo 3 , Rafael 

Bañares 2 ; 1 Innere Medizin I, Martin-Luther-Universität Halle-Wittenberg, 

Halle (Saale), Germany; 2 Digestive Diseases, Hospital Universitario 

Gregorio Marañón, IiSGM. CiberEHD, Madrid, Spain; 

3 Cardiology, Hospital General Universitario Gregorio Marañón, 

IiSGM, Madrid, Spain 

Studies reporting abnormal left ventricular (LV) properties in 

cirrhosis have frequently used load-dependent indices. Aims: 

1) to evaluate systolic and diastolic LV properties with the gold 

standard, LV conductance catheter, which allows obtention 

of real-time pressure volume loops, 2) to test the validity of 

Doppler-echocardiography indices compared to the gold standard, 

3) to analyze the impact of severity of liver disease, 

neurohormonal activation and beta-blocker (BB) treatment on 

LV properties. Methods. Pressure-volume loops were obtained 

in 9 patients with cirrhosis (Child A:3;B:2;C:4; refractory ascites:2) 

in the context of liver transplant evaluation and 9 controls 

undergoing LV catheterization. Invasive gold-standard systolic 

(maximal elastance, Emax) and diastolic indexes (relaxation 

and stiffness) were correlated to Doppler-echocardiography 

indices including peak ejection intraventricular pressure difference 

(EIVPD) and strain rate (SR). Gold-standard validated 

Doppler indexes in cirrhosis (n=59; Child A:15;B:25;C:19; 

refractory ascites:9) were compared to matched controls. 

The influence of the severity of liver disease, neurohormonal 

activation and BB was evaluated. Nonparametric tests were 

used. IRB approval was obtained. Results: Emax correlated 

only with EIPVD (r=0.75, p


1505 

Comparison Of Thrombin Generation After Addition Of 

Thrombomodulin Or Activated Protein C: Looking For A 

Marker Of Hypercoagulability With Low Variability In 

Patients With Cirrhosis 

Cédric Duron 1 , Armando Abergel 1 , Géraldine Lamblin 1 , Aurelien 

Lebreton 2 , Thomas Sinegre 3 ; 1 Hepatogastroenterolgy, CHU, Clermont 

Ferrand, France; 2 CHU Clermont Ferrand, Clermont ferrand, 

France; 3 CHU Clermont Ferrand, Clermont Ferrand, France 

Background and Aims: Cirrhosis is characterized by a defective 

hemostasis, and may result in bleeding complications as 

well as in thrombotic events. Conventional coagulation tests 

such as the Prothrombin Time and aPTT are prolonged in 

patients with cirrhosis, assessing only the defects of procoagulants. 

Thrombin Generation (TG) testing consist in a dynamic 

study of the coagulation, measuring generation and inhibition 

of the thrombin by pro and anticoagulant factors. This new 

approach reveals that TG could be equal or higher in patients 

with cirrhosis than in healthy volunteers. The aim of this study 

was: 1) to assess the variability of TG over time, and 2) the 

effect of Child-Pugh on TG. Methods: Fifty two patients with a 

diagnosis of cirrhosis, without hepatocellular carcinoma, were 

included in the study, in 2 groups: uncomplicated cirrhosis and 

complicated disease defined by the presence of hemorrhage, 

infection or alcoholic hepatitis in the last 3 months. 18 healthy 

volunteers were recruited as controls. Patients or controls using 

drugs known to modify the coagulation system and/or having 

a history of spontaneous thrombosis were excluded. The 

stability of TG in platelet-poor plasma testing, with or without 

addition of thrombomodulin (TM) or activated Protein C 

(PCa), has been assessed over 12 weeks by 3 repeated Calibrated 

Automatic Thrombinography measurements at week 

0, 6 and 12. Results: The intra-individual variability of endogenous 

thrombin potential (ETP) after addition of TM was low, 

about 5%, in uncomplicated cirrhosis and healthy volunteers. 

The inter-individual variability was about 100%, and was not 

influenced by age, sex or cirrhosis etiology (alcoholic or not). 

TG, after addition of PCa, varies over time in cirrhosis and 

healthy groups. In contrast, TG significantly varies over time 

in patients with complicated cirrhosis. TG was significantly 

higher in patients with cirrhosis Child B (n=20) than in Child 

A (n=21) and healthy volunteers (n=18). No difference was 

found between Child B and Child C. The ratio TG without/with 

TM increases with hepatic function. No coagulation imbalance 

persists after in vitro protein C (PC) normalisation. Conclusions: 

TG should be evaluated after addition of thrombomodulin, and 

not after addition of activated Protein C in patients with cirrhosis. 

Thrombin Generation increased according to the hepatic 

function. Thrombin generation over time, in the same patient, 

after addition of thrombomodulin should be evaluated as a risk 

factor of hypercoagulability. Hypercoagulability appears to be 

related to PC deficiency. A largest study should identify factors 

affecting inter and intra individual variabilities. 

Disclosures: 


The following authors have nothing to disclose: Cédric Duron, Géraldine Lamblin, 

Aurelien Lebreton, Thomas Sinegre 

1506 

In Vivo Assessment Of Portal Hypertensive Colopathy 

And Clinical Outcome Of Patients With Liver Cirrhosis 

With Confocal Laser Endomicroscopy (CLE) 

Steffen Zopf 1 , Gian Eugenio Tontini 2 , Michael Vieth 3 , Markus 

F. Neurath 1 , Helmut Neumann 1 ; 1 Medical Department 1, Friedrich-Alexander 

University of Erlangen, Erlangen, Germany; 2 IRCCS 

Policlinico San Donato, San Donato Milanese, Italy; 3 Institute of 

Pathology, Klinikum Bayreuth, Bayreuth, Germany 

Background and Aims: Recent data has highlighted the role 

of mucosal integrity for bacterial translocation in the gut which 

is also discussed as a major cause for development of spontaneous 

bacterial peritonitis (SBP) and/ or hepatic encephalopathy 

(HE) in patients with liver cirrhosis. CLE has emerged as a 

valuable tool for real time diagnosis of mucosal integrity and 

allows in vivo imaging of commensal bacteria in the gut. Our 

aim was to prospectively assess the value of CLE for in vivo 

diagnosis of portal hypertensive colopathy and its association 

to Child-Pugh class, Model for End Stage Liver Disease (MELD), 

HE and development of SBP. Methods: Patients with established 

diagnosis of liver cirrhosis and portal hypertension were 

prospectively included. Clinical, biochemical, and ultrasound 

criteria, including portal vein thrombosis, ascites and collateral 

portosystemic vessels were assessed in addition to endoscopic 

criteria (e.g. esophageal varices, portal gastropathy) and betablocker 

intake. Fluoresceine aided CLE was performed in every 

patient in the sigmoid colon, rectosigmoid junction, and rectum. 

Afterwards biopsies were taken, unless contraindicated, 

for corresponding histopathological analysis. Results: Overall, 

more than 14,700 CLE images were collected. Confocal 

imaging revealed dilation and/or ectasia of microvessels, congestion 

of blood flow, edema, and a non-specific increase of 

the cellular infiltrate within the lamina propria. These findings 

were directly correlated to Child-Pugh class and MELD score 

with patients at higher scores showing more distinct changes 

of the microarchitecture. Of note, disturbed mucosal integrity, 

as observed by CLE, was strongly correlated with occurrence 

of HE and SBP, even in the follow-up of the patients. The procedure 

was well tolerated by the patients, and no adverse 

events were observed. Conclusions: Fluoresceine guided CLE 

in patients with liver cirrhosis and portal hypertensive colopathy 

is safe and well tolerated. In vivo imaging revealed similar 

microscopic changes of portal colopathy as conventional histology 

without the need of physical biopsies. Of note, confocal 

imaging corresponds to clinical outcome parameters, including 

development of HE and/or SPB. 

Disclosures: 

The following authors have nothing to disclose: Steffen Zopf, Gian Eugenio Tontini, 

Michael Vieth, Markus F. Neurath, Helmut Neumann 

1507 

Conjugated bile acids suppress the HPA axis via activation 

of hypothalamic glucocorticoid receptors in a 

rodent model of chronic biliary obstruction 

Matthew McMillin 1,2 , Gabriel A. Frampton 1 , Stephanie Grant 1 , 

Matthew A. Quinn 3 , Sharon DeMorrow 2,1 ; 1 Texas A&M Health 

Science Center, College of Medicine, Temple, TX; 2 Central Texas 

Veterans Healthcare System, Temple, TX; 3 Department of Health 

and Human Services, National Institute of Environmental Health 

Sciences, NIH, Research Triangle Park, NC 

Suppression of the hypothalamic pituitary adrenal axis (HPA) 

occurs during cholestasis. We previously demonstrated that 

serum bile acids gain entry to the hypothalamus in a model 

of cholestasis. The aim of the current study was to determine


the effects of bile acid signaling on the HPA axis. Methods: 

Sprague Dawley rats were fed cholestyramine for 3 days 

prior to sham or bile duct ligation (BDL) surgery; corticosterone 

levels, hypothalamic corticotropin releasing hormone 

(CRH) expression, and serum and hypothalamic bile acid levels 

were assessed. Apical sodium-dependent bile acid transporter 

(ASBT)- and glucocorticoid receptor (GR)-specific Vivo 

morpholinos were infused in the lateral ventricle (1mg/kg/ 

day) for 3 days, followed by injection of the bile acids cholic 

acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid 

(CDCA), taurocholic acid (TCA) or glycochenodeoxycholic 

acid (GCDA) into the 3 rd ventricle (20 pmol) and HPA activity 

was assessed after 6 hr. Hypothalamic neurons were treated 

with various bile acids (10 mM) with or without the GR antagonist 

RU486 or after ASBT knockdown, and CRH expression 

and secretion was assessed. Bile acid uptake was assessed 

with the fluorescent bile acid derivative cholyl-lysyl fluorescein 

(CLF). The ability of bile acids to activate GR was assessed ex 

vivo using a commercially available competition assay and in 

vitro by subcellular localization of GR. Results: Cholestyramine 

attenuated the HPA axis suppression and increase in hypothalamic 

bile acid levels observed during cholestasis. Treatment of 

hypothalamic neurons with various bile acids suppressed CRH 

expression and secretion in vitro. In vivo, HPA axis suppression 

was only evident after injection of conjugated bile acids 

(TCA or GCDA). Uptake of CLF was evident in hypothalamic 

neurons but was inhibited in ASBT-silenced neurons. TCA- or 

GCDA-driven suppression of hypothalamic CRH expression 

and circulating corticosterone could be prevented in vitro and 

in vivo by inhibiting ASBT-mediated bile acid uptake. Both TCA 

and GCDA compete for GR binding sites with endogenous 

ligands in an ex vivo competition assay and caused nuclear 

translocation of GR in vitro. Inhibiting GR activation in vitro 

and in vivo prevented bile acid-driven suppression of the HPA 

axis. Conclusions: Taken together our data support the hypothesis 

that during cholestatic liver injury, bile acids gain entry to 

the brain, are transported into neurons through the ASBT transporter 

and can activate GR to suppress the HPA axis. These 

data also support the broader hypothesis that bile acids may 

act as central modulators of hypothalamic peptides that may be 

altered during liver disease. 

Disclosures: 

The following authors have nothing to disclose: Matthew McMillin, Gabriel A. 

Frampton, Stephanie Grant, Matthew A. Quinn, Sharon DeMorrow 

1508 

Characteristics, indications and outcomes in cirrhotic 

patients with portal hypertension undergoing endoscopic 

retrograde cholangiography: a case-control 

study 

Ricardo Macías-Rodríguez, Jose Luis Rodriguez-Garcia, Astrid 

Ruiz-Margáin, Aldo Torre; Gastroenterology, INCMNSZ, Mexico, 

Mexico 

Background: Endoscopic retrograde cholangiography (ERC) 

is a useful tool for diagnostic and therapeutic biliary tract diseases. 

In patients with cirrhosis, portal hypertension leads to 

clinical and biochemical alterations which can exert higher 

risk for complications related to ERC. However, scarce data 

regarding ERC in cirrhosis are available in medical literature. 

Aim:to evaluate the indications, general characteristics and 

outcomes of ERC in patients with cirrhosis and portal hypertension. 

Methods: This was a case-control study including 37 

patients (71 procedures) with cirrhosis and portal hypertension 

(C) and 37 age and sex-matched controls without cirrhosis 

(NC), undergoing ERC and evaluated 30 days post-ERC. Characteristics 

related to ERC (cannulation, sphincterotomy, stenting 

and complication rate) and cirrhosis (Child-Pugh and MELD 

scores; presence of ascites, hepatic encephalopathy (HE) and 

esophageal varices) were recorded. Mann-Whitney’s U and X 2 

were used as appropriate.Logistic regression and ROC analysis 

with Youden index were used to analyze complications 

in cirrhotics. Results: Main etiology of cirrhosis was hepatitis 

C infection (22%). Patients were classified as Child A/B/C 

(4/18/15), mean MELD was 17.8±6. Complications of cirrhosis 

were ascites (46%), esophageal varices (63%; large 

esophageal varices 43.7%) and HE (16%). Main differences 

in baseline characteristics between groups showed changes 

inherent to cirrhosis in biochemical parameters(higher bilirubin 

and lower PT, platelets, sodium and albumin). Main indication 

in C and NC group for ERC was choledocholithiasis in 46% 

and 48%, respectively p=0.816. Cannulation rate was the 

same in both groups, (97%, p=1.000). Biliary sphincterotomy 

was performed more frequently in NC patients compared to 

C (60 vs 35%, p=0.036); there was no difference in complications 

related to ERC between C and NC (10% vs 8%, 

p=0.677). Factors related to complications in cirrhosis were 

lower alkaline phosphatase levels(230± 71 vs 619± 550mg/ 

dL) and sphincterotomy rate(71 vs 20% for complication and 

no complication subgroup, p=0.010). In the multivariable analysis 

only sphincterotomy was independently associated to the 

presence of complications (OR 9.8[1.7-56.3], p=0.011). ROC 

analysis yielded a MELD score>16 to best predict complications 

after ERC in cirrhosis (Se 71.4%, Sp 57.8%, AUC 0.616). 

Conclusion:Outcomes after ERC in patients with cirrhosis are 

similar to non-cirrhotics despite of the alteration in coagulation 

parameters and the presence of disease-specific complications, 

however a more cautiously approach in patients with cirrhosis 

undergoing sphincterotomy and MELD>16 is needed. 

Disclosures: 

The following authors have nothing to disclose: Ricardo Macías-Rodríguez, Jose 

Luis Rodriguez-Garcia, Astrid Ruiz-Margáin, Aldo Torre 

1509 

Efficacy of Ornithine Phenylacetate (OP) in lowering 

plasma ammonia after upper gastrointestinal bleeding 

(UGIB) in cirrhotic patients: a multicenter, randomized, 

double blind trial 

Meritxell Ventura-Cots 1 , German Soriano 3,4 , Macarena Simon-Talero 

1 , Maria Torrens 1 , Albert Blanco 2 , Jose Antonio Arranz 2 , Mar 

Concepción 3 , Edilmar A. Alvarado 3 , Cristina Gely 3 , Eva Roman 3 , 

Joan Genescà 1,4 , Juan Cordoba 1,4 ; 1 Liver Unit, Hospital Vall d’Hebron, 

Barcelona, Spain; 2 Metabolopathies Unit, Hospital Vall 

d’Hebron, Barcelona, Spain; 3 Department of Gastroenterology, 

Hospital de la Santa Creu i Sant Pau, Barcelona, Spain; 4 Instituto 

de Salud Carlos III, CIBEREHD, Madrid, Spain 

OP has been proposed to decrease ammonia in uncontrolled 

studies. Methods: Thirty-eight consecutive cirrhotic patients, 

aged between 18-85 years and with a creatinine level lower 

than 1.5 mg/dL were enrolled within 24 h of an UGIB. Patients 

were randomized (1:1) to receive OP (10g/24h continuous 

infusion for 5 days) or placebo, in addition to the usual treatment, 

and followed up for 28 days. Ammonia and related 

metabolites were assessed by HPLC and mass spectroscopy. 

The hypothesis was that OP would produce a 25μmol/L venous 

ammonia difference in the means between the groups during 

the first 24h. Secondary outcomes included: plasma ammonia 

changes at any time point, pharmacokinetic profile, hepatic 

encephalopathy and clinical outcome, and confirmation of 

safety and tolerability of OP. Treatment groups were compared 

using a one-sided exact Wilcoxon rank-sum test. Results: a


progressive decrease in ammonia concentration was observed 

in both groups, being higher in OP group at each time point, 

without significant differences. Decrease in ammonia in the first 

24h in the OP group [-20.40 μmol/L (-39.20, -2.80); median 

(IQR)] was twice that of the placebo group [-11.88 μmol/L 

(-36.75, 26.35)], without statistical significance. The subanalysis 

by Child-Pugh score showed a statistically significant 

ammonia decrease in Child-Pugh C treated patients at 36 h, as 

well as in the time normalized area under the curve (TN-AUC) 

0-120h in the OP group [40.16 μmol/L (37.7, 42.6 )] vs placebo 

group [ 65.5 μmol/L (54, 126); p=0.036]. A progressive 

decrease in plasma glycine and glutamine were observed in 

the treated group as compared to the placebo group. These 

differences reached statistical significance after 24 h of treatment 

and were maintained thereafter. Glutamine decrease 

associated with the appearance of phenylacetylglutamine in 

urine: u-PAG accumulated TN-AUC 12-120h placebo [5768 

μmol (1769,8808)] vs TN-AUC 12-120h OP [106851μmol 

(78802, 146666); p50 mmol/L). Ammonemia was higher in patients 

with HE than in those without (108 (95%CI: 86-123) vs 57 

(95%CI: 50-81), p=0.0004). In patients with HE, ammonemia 

was significantly correlated to severity of HE assessed by GCS 

(p=0.001). Overall, 34 HE patients experienced worsening of 

their neurological status during hospitalisation. Ammonemia 

at admission was significantly higher in those patients (128 

(95%CI: 108-147) vs 87 (95%CI: 64-110), p=0.007). In multivariate 

analysis, parameters independently associated with 

worsening of neurological status were MELD score and ammonia 

levels at admission (p=0.01 and 0.02, respectively). Conclusion: 

Ammonia levels at admission are correlated with the 

severity of HE and with outcome in cirrhotic patients with HE. 

Disclosures: 

Marika Rudler - Speaking and Teaching: Gilead, Mayoly 

Thierry Poynard - Grant/Research Support: Gilead; Stock Shareholder: BioPredictive 

Nicolas Weiss - Speaking and Teaching: Norgine, Alpha Wasserman 

The following authors have nothing to disclose: Simona Tripon, Maxime Mallet, 

Denis Monneret, Francoise Imbert-Bismut, Dominique Thabut 

1511 

Use of betablockers, previous hepatic encephalopathy 

and low albumin levels as risk factors of portal vein 

thrombosis in a cohort of cirrhotic patients 

Marta Lopez Gomez 1 , Elba Llop 1 , Angela Puente 2 , Juan de la 

Revilla 1 , Carlos Fernández-Carrillo 1 , Fernando Pons 1 , Jose Luis 

Martinez 1 , Natalia Fernández 1 , Maria Trapero 1 , Javier Crespo 2 , 

José Luís Calleja 1 ; 1 Gastroenterology and Hepatology, Hospital 

Universitario Puerta de Hierro (MADRID. SPAIN), Pozuelo de 

Alarcon, Spain; 2 Hospital Universitario Marqués de Valdecilla, 

Santander, Spain 

Portal vein thrombosis(PVT) is a complication of liver cirrosis(LC). 

The aim of our study was to evaluate anual incidence 

of PVT and related risk factors.Methods: We retrospectively 

reviewed clinical and radiological data collected prospectively 

of consecutive cirrhotic patients included in the database 

of two Universitary Hospitals. Patients out of Milan criteria 

HCC, known PVT, TIPS and pregnancy were excluded. All 

patients with ultrasound diagnosis of PVT underwent MR or 

CTangiography.Results: From September 2013 to September 

2014, 747 cirrhotic patients were reviewed, 179 had exclusion 

criteria. Baseline characteristics are described in Table 

1. 23(4%) patients presented PVT during the inclusion period. 

Significant differences between patients with/without PVT 

were observed in: albumin (3.4SD0.8vs4.0SD0.5;p


HE(OR3.2 IC 1.1-8.; p0.03) were risk factors and high albumin 

levels(OR0.3IC0.2-0.8p=0.01) was as a protective factor. 

Besides, significant differences were observed in PVD(12.2SD- 

5vs10.7SD2;p=0.02) and SD(15SD3vs13SD2.6;p


Room air PaO2, 100% FiO2 PaO2 and MAA are poor predictors 

of overall survival in those considered for MELD exception 

for HPS. Although post-LT mortality is higher in those with a 

100% FiO2 PaO2


1516 

Linkage and interaction between portal hemodynamics 

and serum sodium concentration in cirrhosis 

Hitoshi Maruyama, Takayuki Kondo, Tadashi Sekimoto, Soichiro 

Kiyono, Osamu Yokosuka; Department of Gastroenterology and 

Nephrology, Chiba University Graduate School of Medicine, 

Chiba, Japan 

Background/Aim: Although a hyponatremia is frequently 

observed in patients with cirrhosis, the prognostic influence 

with respect to the presence of concomitant portal hemodynamic 

abnormality has yet to be determined. The study aimed 

to examine the linkage between portal hemodynamics and 

hyponatremia, and their interactions on the prognosis of cirrhosis. 

Methods: This study was based on the subgroup analysis 

using medical records collected in the prospective study 

regarding the influence of portal hemodynamics on clinical 

outcomes from November 2007 to November 2012. Clinical 

findings including portal hemodynamic parameters measured 

by Doppler ultrasound and serum sodium concentrations were 

examined with respect to the prognosis. Following patients 

were excluded: i) with hepatocellular carcinoma (HCC) diagnosed 

by contrast-enhanced CT or magnetic resonance imaging, 

or a treatment history of HCC, ii) receiving medications 

with vasoactive drugs such as β-blockers, antiviral therapy, or 

interventional procedures such as a transjugular intrahepatic 

portosystemic or peritoneo-venous shunt, prior to or during, 

the study period. Results: There were 153 cirrhosis patients 

(62.2 ± 12.0 years; median observation period, 34.1 m). Sixteen 

patients were accompanied with hyponatremia (Na < 

135 mEq/L), who showed a significantly greater frequency of 

possessing a splenorenal shunt (SRS; p = 0.0068), and 137 

patients without hyponatremia. Serum sodium concentrations 

were significantly lower in patients with a SRS than in those 

without (p = 0.0193). An increased prothrombin time-international 

normalized ratio was a significant predictive factor for 

developing hyponatremia a year later (8/96; Hazard ratio 

14.415; p = 0.028). The cumulative survival rate was significantly 

lower in patients with hyponatremia (46.7% at 1 and 

3 years) than in those without (91.8% at 1 year, 76.8% at 3 

years; P < 0.001), and that was significantly lower in patients 

who had developed hyponatremia after one year (100% at 1 

year, 62.5% at 3 years) than those who had not (100% at 1 

year, 89.0% at 3 years; P < 0.001). In addition, the cumulative 

survival rate was significantly worse in patients with both hyponatremia 

and a SRS (20% at one year) than in others (91.8% 

at 1 year, 76.3% at 3 years, and 63.1% at 5 years in patients 

with neither hyponatremia nor a SRS, p < 0.0001; 77.4% at 1 

year, 72.6% at 3 years and 58.1% at 5 years in patients with 

hyponatremia or a SRS, p = 0.014). Conclusions: There was 

a close linkage between the serum sodium concentration and 

portal hemodynamic abnormality, a presence of SRS, and their 

interaction may negatively influence the prognoses in cirrhosis. 

Disclosures: 

The following authors have nothing to disclose: Hitoshi Maruyama, Takayuki 

Kondo, Tadashi Sekimoto, Soichiro Kiyono, Osamu Yokosuka 

1517 

Ammonia Produces Pathological Changes And Activation 

In Human Hepatic Stellate Cells And Is A Target For 

Therapy In Portal Hypertension 

Francesco De Chiara, Rajiv Jalan, Vairappan Balasubramaniyan, 

Fausto Andreola, Massimo Malago, Massimo Pinzani, Rajeshwar 

Mookerjee, Krista Rombouts; UCL, London, United Kingdom 

Background: Hepatic stellate cells (HSC) are vital to hepatocellular 

function and the liver’s response to injury. They share 

a phenotypic homology with astrocytes that are central in the 

pathogenesis of hepatic encephalopathy, a condition in which 

hyperammonemia plays a pathogenic role. This study tested 

the hypothesis that ammonia modulates human HSC activation 

in vitro and in vivo, and evaluated whether ammonia lowering, 

by using L-ornithine phenylacetate (OP), modifies HSC activation 

in vivo and reduces portal pressure in a bile duct ligation 

(BDL) model. Methods: Primary human HSCs were isolated 

and cultured. Proliferation (BrdU), metabolic activity (MTS), 

morphology (TEM, light and immunofluorescence microscopy), 

HSC activation markers by protein analysis, and changes in 

oxidative status (ROS) and endoplasmic reticulum (ER) were 

evaluated to identify effects of ammonia challenge (50 mM, 

100 mM, 300 mM) over 24-72hrs. Changes in plasma ammonia 

levels, markers of HSC activation, portal pressure, hepatic 

eNOS activity and expression of its regulatory proteins were 

quantified in hyperammonemic BDL animals, and after OP 

treatment. Results: Pathophysiological ammonia concentrations 

caused significant and reversible changes in cell proliferation, 

metabolic activity and activation markers of hHSC 

in vitro. Highly significant alterations in cellular morphology, 

characterised by cytoplasmic vacuolisation, ER enlargement 

and ROS production, pro-inflammatory gene expression were 

observed together with HSC-related activation markers such as 

α-SMA, myosin IIa, IIb, and PDGF-Rβ. Treatment with OP significantly 

reduced plasma ammonia (BDL 199.1mmol/L±43.65 

vs. BDL+OP 149.27mmol/L±51.1, P


by, bacterial infections. We recently reported the effects of 

sepsis in the liver microcirculation of healthy rats; however the 

influence of bacterial infections on the liver sinusoidal circulation 

in cirrhotic livers remains unknown. Statins have shown 

beneficial effects ameliorating liver endothelial dysfunction in 

cirrhosis. The present study aimed at characterizing the effects 

of simvastatin on liver microvascular dysfunction in a cirrhotic 

rat model of endotoxemia. Methods: Lipopolysaccharide (LPS; 

1mg/kg) or saline was given to: (1) CCl 4 

-cirrhotic rats treated 

with placebo; (2) cirrhotic rats treated with simvastatin (25 

mg/kg/24 h, orally) from 3 days before LPS/saline injection. 

Hemodynamic parameters (Mean Arterial Pressure-MAP, Portal 

Pressure-PP, Portal Blood Flow-PBF, Hepatic Vascular Resistance-HVR), 

followed by Portal Perfusion Pressure (PPP) and 

microvascular functionality in response to acetylcholine were 

assessed. Underlying molecular/cellular mechanisms (liver 

fibrosis, HSC activation, inflammation, oxidative stress, NO 

release) were investigated. Results: LPS administration markedly 

aggravated hepatic microvascular dysfunction in cirrhotic rats 

(PPP +17%; vasodilatory response to Ach -27%; p


liver sections. Ascitic fluid was evaluated for bacterial growth 

in thioglycolate medium. Ex-vivo liver perfusion experiments 

were performed for assessing endothelium-dependent and – 

independent reactivity. Results: Compared with controls, rats 

with oral CCl4 gavage tended to show decreased survival and 

body weight gain, both of which were further worsened by 

enoxaparin 180 U/kg bw (p< 0.01). Rats with CCL4-induced 

cirrhosis showed altered laboratory parameters (increased 

INR, AST, ALT, bilirubin / decreased albumin, total proteins, 

glucose and platelets) regardless of enoxaparin treatment. In 

all experimental models, cirrhotic rats receiving saline and 

those receiving enoxaparin showed similar increases in the 

area of liver fibrosis compared with controls (p< 0.001). Rats 

with cirrhosis induced by oral CCl4 gavage and by BDL surgery 

developed increases of portal pressure and spleen-to-bw 

ratios compared with control rats (p< 0.001), regardless of 

enoxaparin treatment. Among rats with ascites, a similar proportion 

presented positive bacterial cultures (CCl4+saline 2 

of 7 vs. CCl4+enoxaparin 4 of 8, NS; BDL+saline 1 of 6 vs. 

BDL+enoxaparin 2 of 7, NS). Potential effects of enoxaparin 

on hepatic vascular reactivity were only observed in rats 

receiving enoxaparin 180 U/kg bw from the beginning of 

CCl4 administration, and consisted of increased portal venous 

resistance after addition of acetylcholine or S-nitroso acetylpenicillamine 

(SNAP, both p< 0.05) and increased sinusoidal 

resistance after addition of SNAP (p< 0.05). Conclusion: Our 

experimental data do not support a role of long-term treatment 

with enoxaparin for improving liver fibrosis, portal hypertension 

or endothelial dysfunction in cirrhosis. 

Disclosures: 

The following authors have nothing to disclose: Jose Ignacio Fortea, Alexander 

Zipprich, Carolina Fernandez Mena, Christopher F. Rose, Juan Bañares, Marta 

Puerto, Cristina R. Bosoi, Jorge Almagro, Marcus Hollenbach, Marc-André Clément, 

Javier Vaquero, Rafael Bañares, Cristina Ripoll 

1521 

Neuro-inflammation in Cirrhotic Mice is Dependent on 

Gut Microbial Colonization: Implications for Hepatic 

Encephalopathy 

Dae Joong Kang 1 , Siddhartha Ghosh 1 , Daniel Carl 1 , Arun J. 

Sanyal 1 , Ryan B. Sartor 2 , Phillip B. Hylemon 1 , Huiping Zhou 1 , Runping 

Liu 1 , Xiang Wang 1 , Jing Yang 1 , Chunhua Jiao 1 , Jasmohan S. 

Bajaj 1 ; 1 VCU and McGuire VAMC, Richmond, VA; 2 Gnotobiotic 

Rodent Research Center, University of North Carolina, Chapel 

Hill, NC 

Neuro-inflammation has been proposed as a mechanism for 

hepatic encephalopathy (HE) but the source for this is not clear. 

Specifically, brain inflammation in cirrhotic models in the germfree 

mice (GF) condition has not been studied. Aim: Assess the 

impact of intestinal microbiota on brain inflammation in cirrhotic 

mice in a conventional (conv) and GF setting. Methods: 

We used C57BL/6 mice in a conv and GF setting. Sixteen mice 

were in the GF condition; of which 6 remained GF while 10 

were administered CCL4 1ml/kg in sesame oil using gavage 

twice/week till week 16. An age-balanced group of 10 conv 

mice were included; 5 remained healthy while cirrhosis using 

CCL4 was induced in 5 mice using 1ml/kg CCL4 administration 

twice a week for 12 weeks. On sacrifice, brain tissue was 

separated into cortex (Cx) and cerebellum (Cbl). Analysis was 

performed for mRNA of inflammation (IL-6, IL-1β) and activation 

of microglia and glia (IBA-1, GFAP) using qPCR (adjusted for 

GADPH) in both brain regions of all mice using Kruskall-Wallis 

tests. Results: All mice survived till end of the experiment. Both 

CCL4-treated groups (conv & GF) showed cirrhosis on sacrifice 

on visual and histological examination. Comparison within GF 

group: There was no significant difference in inflammation or 

microglial/glial activation in both brain regions in GF controls 

vs. CCL4 GF cirrhotics (table) Comparison within Conv 

group: There was a significant increase in mRNA of IL-1β, 

GFAP and IBA-1 in both Cbl and Cx of CCL4 conv cirrhotic 

mice compared to their conv non-cirrhotic counterparts. IL-6 

was unchanged. Comparison between GF Cirr and Conv cirr: 

Conv CCL4 cirrhotic mice showed a significant increase in Cbl 

and Cx IL-1β, GFAP and IBA-1compared to GF cirrhotic mice. 

IL-6 was again not different between groups. Conclusions: Glial 

and microglial activation and induction of IL-1β occurs in cirrhotic 

mice only in the presence of gut microbiota. This suggests 

an essential role of the gut microbiota in the development 

of neuro-inflammation in cirrhosis and could have implications 

for the gut-brain axis management in hepatic encephalopathy. 

Neuroinflammation and Glial/Microglial Expression between 

Groups 

Disclosures: 









The following authors have nothing to disclose: Dae Joong Kang, Siddhartha 

Ghosh, Daniel Carl, Ryan B. Sartor, Phillip B. Hylemon, Huiping Zhou, Runping 

Liu, Xiang Wang, Jing Yang, Chunhua Jiao 

1522 

Emricasan, a pan caspase inhibitor, improves survival 

and portal hypertension in a murine model of long-term 

common bile-duct ligation 

Akiko Eguchi 1 , Alexander Wree 1 , Yukinori Koyama 1 , Casey Johnson 

1 , Ryota Nakamura 1 , Patricia C. Contreras 2 , Ariel E. Feldstein 1 ; 

1 UCSD, La Jolla, CA; 2 Conatus Pharmaceuticals, San Diego, CA 

Development of portal hypertension (PTH) is a central prognostic 

factor in patients with cirrhosis. Current pharmacotherapy 

remains limited and novel therapies are greatly needed. Circulating 

microparticles (MPs) are released by hepatocytes in 

a caspase dependent manner, are increased in circulation of 

patients with cirrhosis and contribute to PTH via induction of 

impair vasoconstrictor responses Here we tested the hypothesis 

that Emircasan (IDN-6556) a pan-caspase inhibitor ameliorates 

PTH via its anti-fibrotic effects and reduction in release 

of MPs. Methods: Secondary biliary cirrhosis was induced by 

long-term common bile-duct ligation (CBDL) in C57BL/6 mice. 

Controls were sham-operated, the abdominal cavity opened, 

but no ligature placed. Mice were treated with 10 mg/kg/ 

day of IDN-6556 or placebo for 3 wks via i.p. injection (4 

groups, n= 7 -12 in each group).. After 3 wks, portal pressure 

was measured in each mouse and serum and livers were collected. 

Circulating MPs were isolated and characterized using 

various approaches including FACS analysis. Portal pressure 

was measured using catheter from ileocolic vein. Hepatocellular 

damage was assessed by liver histopathology, serum ALT 

levels, and TUNEL assay. Liver fibrosis was assessed by digital 

image analysis of Sirius red stained sections. Results: In


contrast to CBDL-Placebo group, nearly all CBDL-IDN-6556 

survived (p


HVPG ≥10mmHg (R=0.818, p=0.007). Conclusion We have 

demonstrated that parameters related to both hepatic architecture 

and splanchnic haemodynamics derived from non-contrast 

quantitative MR imaging techniques correlate significantly with 

HVPG. 

Disclosures: 

The following authors have nothing to disclose: Naaventhan Palaniyappan, Eleanor 

Cox, Andrew Austin, Indra Neil Guha, Susan Francis, Guruprasad P. Aithal 

1525 

The soluble guanylyl cyclase stimulator riociguat 

reduces liver fibrosis and portal pressure in cirrhotic rats 

Philipp Schwabl 1 , Ksenia Brusilovskaya 1 , Florian Riedl 1 , David 

J. Bauer 1 , Bastian Strobel 1 , Paul Supper 1 , Hubert Hayden 1 , 

Michael Trauner 1 , Bruno K. Podesser 2 , Katharina Wochner 2 , 

Markus Peck-Radosavljevic 1 , Thomas Reiberger 1 ; 1 Division of Gastroenterology 

and Hepatology, Department of Internal Medicine 

III, Medical University of Vienna, Vienna, Austria; 2 Department 

for Biomedical Research, Medical University of Vienna, Vienna, 

Austria 

Background: In liver cirrhosis nitric oxide (NO) signaling as 

well as the function of its key downstream target soluble guanylyl 

cyclase (sGC) is distorted. The sGC stimulator riociguat 

(RIO) is used for treatment of pulmonary hypertension. Since 

RIO has been shown to have additional antifibrotic activity, 

we aimed to investigate effects of RIO in rats with cirrhotic 

portal hypertension. Methods: Cirrhosis was induced by carbontetrachloride 

injections (CCl4, 8 weeks) or by bile duct ligation 

(BDL, 3 weeks) in Sprague Dawley rats. Controls received 

olive-oil (OO) or underwent sham operation (SO), respectively. 

RIO (1mg/kg/d, gavage) or vehicle (VEH) was administered 

from weeks 6-8 in CCl4/OO and from weeks 2-3 in BDL/ 

SO animals. Mean arterial pressure (MAP), heart rate (HR), 

portal pressure (PP) and superior mesenteric artery blood flow 

(SMABF) were measured. Porto-systemic and spleno-renal shunting 

was assessed by colored microspheres technique. Hepatic 

fibrosis was quantified by hepatic hydroxyproline content (HP) 

and histology. Results: CCl4 rats presented with marked cirrhosis, 

hyperdynamic circulation, portal hypertension and elevated 

SMABF compared to OO animals. In CCl4 rats, RIO had no 

effect on HR, MAP and PP but significantly decreased SMABF 

(74±11 vs. 43±6 mL/min; p=0.036). Further, in CCl4 cirrhosis 

RIO significantly decreased spleen (1.3±0.1 vs. 1.0±0.1g; 

p=0.008) and liver (16.7±1.2 vs. 11.5±2.4g; p=0.015) 

weight. BDL rats presented with hepatosplenomegaly and portal 

hypertension. In the BDL model RIO significantly increased 

MAP (85±17 vs. 108±14mmHg; p=0.016) and decreased PP 

(13.2±2.5 vs. 10.1±2.4; p=0.048), but had no effect on HR, 

SMABF or shunting. HP was significantly decreased (286±147 

vs. 144±74 μg; p=0.039) in BDL-RIO rats, compared to BDL- 

VEH animals. Conclusions: RIO treatment significantly reduced 

liver fibrosis and ameliorated hyperdynamic circulation as well 

as portal pressure in biliary cirrhosis. In toxic cirrhosis RIO 

prevented development of hepatosplenomegaly and reduced 

splanchnic blood inflow compared to controls. RIO could be 

a good candidate for combination studies with other agents 

known to improve hepatic hemodynamics in cirrhosis of various 

etiologies. 

Disclosures: 











The following authors have nothing to disclose: Philipp Schwabl, Ksenia Brusilovskaya, 

Florian Riedl, David J. Bauer, Bastian Strobel, Paul Supper, Hubert 

Hayden, Bruno K. Podesser, Katharina Wochner 

1526 

Portal tract fibrosis is induced by hepatic arterial buffer 

response in portal hypertensive rats: A role for macrophage 

miR154 released via exosome 

Li Gong 1,2 , Chao Dong 1,3 , Teruo Utsumi 1 , Yasuko Iwakiri 1 ; 1 Internal 

Medicine, Section of Digestive Diseases, Yale University School 

of Medicine, New Haven, CT; 2 Anesthesiology, The third Xiangya 

Hospital, Central South University, Changsha, China; 3 General 

Surgery, Xiangya Hospital, Central South University, Changsha, 

China 

Background: The hepatic arterial buffer response increases 

hepatic arterial (HA) flow in response to decreased portal 

venous flow in portal hypertension. Hemodynamic stresses 

associated with increased HA flow may influence intrahepatic 

microenvironments around the HA, including resident fibroblasts 

of the portal tract region, portal myofibroblast (PMF), 

which is a major contributor to portal tract fibrosis. The relation 

between increased HA flow and portal tract fibrosis has 

not been determined. We hypothesize that hemodynamic 

stresses associated with increased HA flow promote infiltration 

of macrophages that modulate PMF function and contribute 

to the development of portal tract fibrosis in portal hypertension. 

Method: Male Sprague Dawley rats underwent sham 

operation or partial portal vein ligation (PPVL) to induce portal 

hypertension. Livers were isolated 10 days after the surgery 

for histological, immunohistochemical and biochemical analyses. 

Microarray analysis was performed to determine changes 

in gene profiling between sham and PPVL rats. To examine 

exosome transfer from macrophages to PMF, co-culture experiments 

were performed using macrophage-rich cell population 

isolated from spleens of rats that express green fluorescent 

protein (GFP) and PMF of normal rats. Exosomes were verified 

by scanning electron microscope, CD63 (an exosome marker), 

and nanoparticle tracking analysis. Results: Sirius red staining 

of livers determined that portal tract fibrosis significantly 

developed in rats 10 days after PPVL (2-fold, p


Hepatic arterial buffer response induces portal tract fibrosis in 

portal hypertension. miR154 in exosomes released from macrophages 

to PMF may play a role in PMF proliferation and 

contribute to portal tract fibrosis 

Disclosures: 

The following authors have nothing to disclose: Li Gong, Chao Dong, Teruo 

Utsumi, Yasuko Iwakiri 

1527 

Diabetes is Associated with Stool and Sigmoid Mucosal 

Microbiota Dysbiosis Independent of MELD score in Cirrhosis: 

Implications for Prognostication 

Jasmohan S. Bajaj 1 , Naga Betrapally 2 , Douglas M. Heuman 1 , Melanie 

White 1 , Kalyani Daita 1 , Ariel Unser 1 , Edith A. Gavis 1 , Phillip 

B. Hylemon 1 , Swati S. Almeida-Dalmet 2 , Masoumeh Sikaroodi 2 , 

Patrick M. Gillevet 2 ; 1 VCU and McGuire VAMC, Richmond, VA; 

2 Microbiome Analysis Center, George Mason University, Manassas, 

VA 

Cirrhotics have an unfavorable gut microbial profile (dysbiosis) 

that is linked to a pro-inflammatory milieu and several 

complications. Type 2 diabetes(DM) can increase the risk of 

complications but the mechanisms are unclear. Aim: Define 

the effect of DM on gut microbiota in cirrhotics and its linkage 

with cirrhosis and DM severity.Methods: Cirrhotic outpts (with/ 

without DM) underwent stool & sigmoid mucosal microbiota 

analysis (multi-tag pyrosequencing). DM severity (insulin use, 

HgbA1c within 6 mths) & cirrhosis severity (MELD) data were 

collected. The cirrhosis dysbiosis ratio (CDR: Lachnospiraceae 

+ Ruminococceae + Clostridiales XIV + Veillonellaceae/Enterobacteriaceae 

+ Bacteroidaceae, low score is worse) ratio of 

commensal vs. potentially pathogenic taxa in stool was calculated. 

LFSe analysis with multiple comparisons adjustment for 

stool/mucosal microbiota were performed between cirrhotics 

with/without DM & those on/not on insulin. Results: 287 cirrhotics 

(57yrs, MELD 11, 39% hepatic encephalopathy (HE), 

40% HCV, 17% NASH) gave stool while 72 cirrhotics (29% 

DM, 36% HE) underwent sigmoidoscopy. 30% (n=87) of cirrhotics 

had DM (45% on insulin, HgbA1c 6.9). MELD & HE% 

was similar but DM pts had higher %NASH & BMI (Table). 

Microbiome analysis: Stool: showed significant dysbiosis in 

DM with a low CDR. This was due to a higher Bacteroidaceae 

(29% vs 14%, p=0.04) &Streptococcaceae (0% vs 2%, 

p=0.02) with lower commensal abundance. The lower bacteria 

in DM were Ruminococcaeae (3% vs 8%, p


1529 

Elevated galectin-3 in cirrhotic heart increases TNFα 

and inhibits cardiac contractility in rats 

Hongqun Liu, Samuel S. Lee; university of calgary, Calgary, AB, 

Canada 

Background: Galectin-3 plays a key pathogenic role in heart 

failure. However, the role of galectin-3 in cirrhotic cardiomyopathy 

is unclear. The present study aimed to investigate the 

effects of galectin-3 on cardiomyopathy. Methods: Two sets of 

rats were used in this study, one to study structural changes and 

another for a functional study. There were 4 groups (n=6 in 

each group) in each set: bile duct ligation (BDL); BDL + galectin-3 

inhibitor (N-acetyllactosamine, LacNAc, 5 mg/kg); sham 

operation and sham + LacNAc. The rats were sacrificed 4 

weeks after surgery. Cardiac mRNA and protein contents were 

evaluated by RT-PCR and Western blots (Galectin-3, PKCα, 

fibronectin, collagen I and III). Heart and serum TNFα was 

measured by ELISA. Isolated cardiomyocyte contractility was 

measured by an IonOptix cell-shortening video system. Results: 

the galectin-3, fibronectin and collagen I/ III ratio were significantly 

increased in cirrhotic hearts. TNFα was significantly 

increased in BDL rats in both serum and heart compared with 

sham controls (744 ± 84 pg/ml vs 566 ± 61 pg/ml in serum, 

p< 0.01 and 310 ± 32 pg/mg protein vs 189 ± 21 pg/ 

mg protein in heart, p


Adipose triglyceride lipase (ATGL) is the rate-limiting enzyme 

for triglyceride hydrolysis in adipose tissue, muscle, and liver. 

In a murine model of cancer cachexia, global deletion of ATGL 

prevented tumor-induced weight loss by inhibiting both adipose 

tissue and skeletal muscle wasting. Whether tissue-specific 

ATGL inactivation can ameliorate chronic-liver disease 

associated cachexia is unknown. Our aim was to determine 

the role of adipose tissue ATGL on cachexia development in 

a murine model of biliary injury (bile duct ligation). Methods: 

Adipose-tissue specific ATGL-knockout (AAKO, N = 10) and 

wild type C57/BL6 (N = 10) male mice 10-15 weeks old were 

treated with bile duct ligation (BDL) or sham laparotomy. Calorie 

intake and body weight were measured weekly. Body composition 

was determined weekly using Echo-MRI, and 36-hour 

respiratory exchange ratio (RER) and activity were measured 

in an Oxymax Comprehensive Laboratory Animal Monitoring 

System (CLAMS). Animals were sacrificed 4 weeks after surgery. 

Results: At baseline, AAKO mice exhibited increased 

adipose tissue mass compared to WT mice. BDL-treated WT 

and AAKO mice developed moderate fibrosis by Sirius Red 

staining. There were no differences in caloric intake between 

genotypes and treatment groups. BDL significantly reduced fat 

mass and lean mass (% total body weight) in AAKO mice 

only (p


values of the spleen was 0.84. VS values of the spleen were 

significantly lower in the group of patients with F2 varices with 

the presence of the major shunt. Conclusion; In patients with 

liver cirrhosis, Vs values of the spleen by VTQ were useful in 

the detection of EGV. They were extremely useful in particular 

in detecting esophageal and gastric varices which require 

treatment. 

Disclosures: 

The following authors have nothing to disclose: Chikage Nakano, Takashi 

Nishimura, Tomoko Aoki, Kyohei Kishino, Yoshihiro Shimono, Kunihiro Hasegawa, 

Ryo Takata, Kazunori You, Akio Ishii, Tomoyuki Takashima, Yoshiyuki 

Sakai, Nobuhiro Aizawa, Naoto Ikeda, Hiroki Nishikawa, Yoshinori Iwata, 

Hirayuki Enomoto, Shuhei Nishiguchi, Hiroko Iijima 

1534 

All-oral direct acting antivirals can offer significant cost 

savings and cost effectiveness for treating hepatitis C 

in medically underserved areas in the United States: 

Impact of task-shifting on access 

Channa R. Jayasekera 1 , Nathaniel Smith 2 , Ryan B. Perumpail 1 , 

Robert J. Wong 3 , Rachel Beckerman 2 , Zobair M. Younossi 4,5 , 

Aijaz Ahmed 1 ; 1 Division of Gastroenterology and Hepatology, 

Stanford University Medical Center, Stanford, CA; 2 CB Partners, 

New York, NY; 3 Division of Gastroenterology and Hepatology, 

Alameda County Health System - Highland Hospital, Oakland, CA; 

4 Center for Liver Diseases, Inova Fairfax Hospital, Falls Church, 



Purpose: The superior safety and tolerability of all-oral 

direct-acting antivirals (DAAs) for chronic hepatitis C (HCV) 

allows specialist physicians to devolve treatment monitoring to 

mid-level healthcare providers, particularly in areas of health 

worker shortage (task-shifting). Based on our task-shifting experiences 

in rural clinics, we used a decision-analytic Markov 

model to assess 1) budgetary impact and 2) cost-effectiveness 

of task-shifted HCV treatment with second generation all-oral 

DAAs (2nd Gen DAA, e.g. ledipasvir/sofosbuvir) vs. first generation 

DAA (e.g. boceprevir)+peginterferon+ribavirin (PR+1st 

Gen DAA) Methods: We modelled a hypothetical cohort of 

1000 patients with HCV genotype 1 with demographics mirroring 

those in our rural clinics, over a lifetime horizon. Cost of 

treatment monitoring, cost per sustained virological response 

(SVR), cost per patient, and overall budget impact were compared. 

Cost-effectiveness of 2nd Gen DAA over PR+1st Gen 

DAA were evaluated from a US third-party payer perspective, 

using published clinical trial and real world SVR rates for the 

regimens. Treatment with 2nd Gen DAA was assumed to double 

treatment capacity; that is 1,000 patients receiving 2nd 

Gen DAA (intervention arm), 500 patients receiving PR+1st 

Gen DAA, and 500 untreated patients (comparator arm). 

Transition probability, utility, and cost estimates (2014 dollars) 

were based on literature and hepatologists consensus. 

Results: Driven by limited monitoring, shorter treatment duration, 

and lower reimbursement rates, 2nd Gen DAA treatment 

monitoring by mid-level providers led to reductions of 74% in 

monitoring costs, 38% in cost per SVR, and 25% in total cost 

of treatment compared with PR+1st Gen DAA. Assuming a willingness-to-pay 

threshold of $50,000 per quality-adjusted life 

year (QALY), 2nd Gen DAA treatment monitoring by mid-level 

providers demonstrated higher efficacy and lower total costs 

(i.e. a dominant strategy). Conclusions: In this model based 

on rural clinic populations, task-shifting of 2nd Gen DAA treatment 

monitoring from specialists to mid-level providers led to 

significant cost savings and was cost-effective. These findings 

support task-shifting as a potential strategy to overcome significant 

gaps in access to all-oral DAA regimens in medically 

underserved areas in the United States. 

Disclosures: 

Nathaniel Smith - Consulting: Gilead Sciences 

Rachel Beckerman - Consulting: Gilead Sciences 






Inc. 

The following authors have nothing to disclose: Channa R. Jayasekera, Ryan B. 

Perumpail, Robert J. Wong 

1535 

Inappropriate Laboratory Utilization: Acute Viral Hepatitis 

Serologies 

Kamran Hussaini 1 , Muhammad B. Hammami 1 , Edward Charbek 

4,1 , Robin Chamberland 2 , Hamza Khalid 3,1 , Brent A. Neuschwander-Tetri 

3,1 ; 1 Internal Medicine, Saint Louis University, St. 

Louis, MO; 2 Pathology, Saint Louis University, St. Louis, MO; 3 Gastroenterology 

& Hepatology, Saint Louis University, St. Louis, MO; 

4 Pulmonary, Critical Care & Sleep Medicine, Saint Louis University, 

St. Louis, MO 

Purpose: Inappropriate laboratory tests contribute to excessive 

health care costs. We have observed acute viral hepatitis serologic 

testing in patients with chronic liver disease, sometimes 

repetitively, in the absence of substantially elevated aminotransferases. 

Our aim was to determine the rate of unnecessary 

testing for acute hepatitis A (HAV) and B (HBV) infections in 

our institution and estimate the impact on health care costs. 

Methods: The results of serum anti-HAV IgM and anti-HBc IgM, 

and alanine aminotransferase (ALT) and aspartate aminotransferase 

(AST) were collected from the Saint Louis University Hospital 

Electronic Medical record systems, Epic and Meditech, 

between January 2010 and Nov 2014. Due to an error in data 

collection no data was collected for June to December 2013. 

The highest value of ALT or AST within the 30 days before testing 

anti-HAV IgM and anti-HBc IgM were extracted. Serological 

tests without ALT and AST values were excluded. Results were 

categorized as shown in the table below; ALT or AST level > 

100 Units/L was chosen to be the lowest level appropriate for 

ordering the serologies. The cost of inappropriate testing was 

estimated based on Medicare reimbursement of $15.32 for 

anti-HAV IgM and $58.92 for anti-HBc IgM. Results: Anti-HAV 

IgM and anti-HBc IgM were tested a total of 2439 and 4462 

times, respectively over the approximately 5 year period. As 

shown in the table below, 72.7% of anti-HAV IgM and 82.7% 

of anti-HBc IgM tests were inappropriate. Medicare payment 

would have been on average $5,435 per year (3.37% of 

charges) for anti-HAV IgM and $43,494 per year (13.57% 

of charges) for anti-HBc IgM. If the threshold for inappropriate 

testing is set at an ALT and AST < 250 U/L, then 90.7% of anti- 

HAV IgM and 93.7% anti-HBc IgM tests were inappropriate. 

The actual health care costs may be higher based on higher 

reimbursement rates from commercial insurers. Conclusions: 

Most testing for acute viral hepatitis is ordered inappropriately. 

We predict this is a nationwide practice that needs to be 

addressed through education and other interventions that might


include warnings provided in the electronic ordering system 

and instituting a policy for laboratory personnel to confirm the 

test indication with the ordering provider before collecting and 

processing the specimen. 

Disclosures: 

Robin Chamberland - Advisory Committees or Review Panels: Bacterioscan, Inc. 



Scholar Rock 

The following authors have nothing to disclose: Kamran Hussaini, Muhammad B. 

Hammami, Edward Charbek, Hamza Khalid 

1536 

Cost-effectiveness analysis of sofosbuvir plus ledipasivir 

in the treatment of chronic hepatitis C virus genotype 1 

infection in real life practice in China 

Bing Li 1 , Jing Chen 2 , Cheng Wang 2,3 , Vanessa Wu 2 , April Wong 2 , 

Qing Shao 1 , Dong Ji 1 , Fan Li 1 , Yudong Wang 2 , Xiaoxia Niu 1 , Jialiang 

Liu 1 , Wucai Yang 1 , Yiming Fu 1 , Guofeng Chen 1 , George K. 

Lau 2 ; 1 Liver Fibrosis Diagnosis and Treatment Center, 302 Hospital, 

Beijing, China; 2 Gastroenterology & Hepatology, Humanity & 

Health Medical Group, Hong Kong, China; 3 Department of Infectious 

Diseases and Liver Unit, Nanfang Hospital, Southern Medical 

University, Guangzhou, China 

Background and Aims: China has about 50 million individuals 

infected with chronic hepatitis C virus (HCV), among which 

genotype 1 accounts for almost 60%. New interferon (IFN)- 

free regimens to treat HCV genotype 1 are more effective than 

the standard of care (SOC) but substantially more expensive. 

Sofosibuvir (SOF), for example, has demonstrated high efficacy 

in patients with HCV genotype 1, 2, 3, or 4 infection. We 

aimed to evaluate the cost-effectiveness of 12-week sofosibuvir 

(Sovaldi®, Gilead) plus daclatasvir (Daklinza®, BMS) (SOF/ 

DCV) compared to SOC in treatment-naïve HCV genotype 1b 

patients in real life practice in Chinese. Methods: A Markov 

transition model was constructed based on the natural history 

of HCV infection from the perspective of third-party payer. A 

cohort of patients were stratified by sex and cirrhosis stages 

and followed from age 50 to simulate the lifetime cost of HCV 

infection and IFN-free treatment as well as the health outcome 

in quality adjusted life years (QALYs). SVR rates were from 

real life data. Utility scores for QALYs and transition rates 

among different stages of HCV were from literature. Direct 

medical costs in China were used and costs were in US$. Both 

costs and health outcome were discounted at 3%. One-way 

sensitivity analysis and second order Monte Carlo simulation 

were undertaken to assess parameter uncertainty. Results: The 

SOF/LDV regimen added 0.84 QALY compared to SOC, at 

an incremental cost effectiveness ratio (ICER) of US$68,397 

per additional QALY. The ICERs ranged from US$108,672 to 

US$3,280 with respect to sex and presence of cirrhosis. The 

ICERs were lower in patients with cirrhosis than those without 

cirrhosis. One-way sensitivity analysis showed that the results 

were most sensitive to utility scores after successful treatment, 

cost of SOF/LDV treatment, discount rate, and transition probability 

of decompensate cirrhosis to death status. At a willingness 

to pay threshold of US$40,842 (6 times GDP per capita 

in China) per QALY, SOF/LDV regimen had a 58% probability 

of being cost-effectiveness. Threshold analysis showed that the 

cost of SOF/LDV treatment needs to be reduced by at least 

25% to make the regimen cost-effective in Chinese patients. 

Conclusion: Though the efficacy is significantly improved using 

IFN-free regimen compared to SOC, the IFN-free regimen is 

not cost-effective in most Chinese HCV genotype 1b patients 

mainly due to the high cost of IFN-free regimen and relatively 

low GDP per capital in China. Reduce the cost and shorten the 

duration of treatment may improve the cost-effectiveness in the 

future. 

Disclosures: 

George K. Lau - Consulting: Roche, Novartis, Roche, Novartis, Roche, Novartis, 

Roche, Novartis 

The following authors have nothing to disclose: Bing Li, Jing Chen, Cheng Wang, 

Vanessa Wu, April Wong, Qing Shao, Dong Ji, Fan Li, Yudong Wang, Xiaoxia 

Niu, Jialiang Liu, Wucai Yang, Yiming Fu, Guofeng Chen 

1537 

Cost-effectiveness analysis of sofosbuvir-based regimens 

for chronic hepatitis C virus genotype 1 infection 

in China 

Fan Li 1 , Guofeng Chen 1 , Qing Shao 1 , Dong Ji 1 , Bing Li 1 , Zhongbin 

Li 1 , Jialiang Liu 1 , Chunxiao Liu 1 , Tingting Wang 1 , Qiaomin 

Wang 1 , Jing Chen 2 , Vanessa Wu 2 , April Wong 2 , Cheng Wang 2 , 

Yudong Wang 2 , George K. Lau 1,2 ; 1 Liver Fibrosis Diagnosis and 

Treatment Center, 302 Hospital, Beijing, China; 2 Gastroenterology 

& Hepatology, Humanity & Health Medical Group, Hong 

Kong, China 

Background and Aims: The nucleotide polymerase inhibitor 

sofosbuvir (SOF) has demonstrated high sustained virologic 

response (SVR) rates in treatment-experienced HCV infection 

but very costly. We aimed to evaluate the cost-effectiveness of 

sofosbuvir (SOF)-based regimens i.e. 12-week SOF (Sovaldi®, 

Gilead) plus peginterferon/ribavirin (SOF+PEG/RBV) and 

12-week SOF (Sovaldi®, Gilead) plus Daclatasvir (Daklinza®, 

BMS) (SOF+DCV) compared to SOC in treatment-experienced 

HCV genotype 1b patients in Chinese. Methods: A Markov 

model with a lifetime, third-party payer’s perspective was constructed 

to simulate the disease progression of a cohort of 50 

year old HCV patients. The patients were stratified by sex and 

cirrhosis status. Outcome measures included quality adjusted 

life years (QALYs), lifetime costs of HCV infection and incremental 

cost-effectiveness ratios (ICERs). Both cost and health 

outcome were discounted at 3%. The robustness of the results 

was tested by one-way sensitivity analysis and probabilistic 

sensitivity analysis. Results: Compared to SOC, SOF+DCV 

were cost-effective in patients with cirrhosis with an ICER of 

US$13,304 (15,842 and 10,233 for males and females 

respectively)(Table). SOF-based treatments were not cost-effective 

in patients without cirrhosis. The cost-effectiveness of SOFbased 

treatments was better in females than males across all 

types of patients. The results were sensitive to post SVR utility, 

costs of SOF-based treatments, and discount rate. At the WTP 

threshold of US$20,421 (3 times of GDP per capita in China), 

SOF+PEG/RBV were cost-effective in 88% and 97% non-cirrhotic 

and cirrhotic patients; SOF+DCV were cost-effective in 

40% and 98% non-cirrhotic and cirrhotic patients. Conclusion: 

In treatment-experienced HCV patients, SOF-based treatments 

may be a cost-effective alternative to SOC depending on the 

costs of SOF-based drugs.


Disclosures: 

George K. Lau - Consulting: Roche, Novartis, Roche, Novartis, Roche, Novartis, 

Roche, Novartis 

The following authors have nothing to disclose: Fan Li, Guofeng Chen, Qing 

Shao, Dong Ji, Bing Li, Zhongbin Li, Jialiang Liu, Chunxiao Liu, Tingting Wang, 

Qiaomin Wang, Jing Chen, Vanessa Wu, April Wong, Cheng Wang, Yudong 

Wang 

1538 

The Value of Cure Associated with Treating Treatment-naïve 

(TN) Chronic Hepatitis C (CH-C) Genotype 1 

(GT1): Are the New All Oral Regimens a Good Value to 

Society? 

Zobair M. Younossi 1,2 , Haesuk Park 3 , Douglas Dieterich 4 , Sammy 

Saab 5 , Aijaz Ahmed 6 , Stuart C. Gordon 7 ; 1 Center For Liver 



Inova Health System, Falls Church, VA; 3 University of Florida, 

Gainesville, FL; 4 Icahn School of Medicine at Mount Sinai, New 

York City, NY; 5 University of California Los Angeles, Los Angeles, 

CA; 6 Stanford University, Standford, CA; 7 Henry Ford Hospital, 

Detroit, MI 

Background and Aim: The approval of all-oral regimens has 

led to substantially high cure rates. Our aim was to assess the 

value of cure to society by treating HCV GT1. Methods: A Markov 

model for HCV GT1 projected long-term health outcomes, 

life-years (LYs), and quality-adjusted life-years (QALYs) gained 

for a lifetime horizon. The model compared current therapies in 

the US [2nd generation triple (sofosbuvir+PR & simeprevir+PR) 

and all-oral therapy treatments (ledipasvir/sofosbuvir and 

ombitasvir+paritaprevir/ritonavir+dasabuvir±ribavirin] with 

no treatment option. Sustained virologic response (SVR) rates 

were based on Phase III clinical trials. We assumed that 80% 

and 95% of HCV GT1 patients are eligible for 2nd generation 

triple and all-oral regimens. Transition probabilities, utility and 

mortality were based on literature review and consensus by 

hepatologists. Results: In 2015, 1.11 million individuals were 

TN with HCV-GT1. The model estimated that treating all eligible 

HCV GT1 patients with 2nd generation triple and all-oral 

therapies can result in 1.8 million and 2.9 million additional 

QALYs gained. Using a threshold of $50,000 to $150,000 

for the value of a QALY, these regimens led to savings of $91- 

$274 billion and $144-$432 billion. Also, the costs of these 

regimens were $80 billion and $93 billion. The value of cure 

was assessed by subtracting the costs of the regimens from the 

economic gains associated with the value of QALYs. The value 

of cure with 2nd generation triple and all oral regimens was 

estimated at $12-$194 billion and $51-$340 billion. Each 

HCV GT1 patient cured with 2nd generation triple and all oral 

regimens could save the U.S. between $10,381-$174,473 

and $46,141-$305,021. Conclusions: The recent evolution 

of regimens for HCV GT1 has increased efficacy and value of 

cure. Curing all eligible HCV-GT1 patients with all-oral regimen 

leads to substantial savings to the U.S. society. 

Disclosures: 



Haesuk Park - Consulting: Gilead Science 










Inc. 




1539 

Rethinking pricing policy of direct antiviral agents to 

cure chronic hepatitis C infection at a population-level 

Francois Girardin, Nathalie Vernaz, Nicolas Goossens, Francesco 

Negro; Geneva University Hospital, Geneva, Switzerland 

Background Although cost-effectiveness of new direct antiviral 

agents for chronic hepatitis C viral infection (HCV) is a 

prerequisite for reimbursement, a population-level cure would 

induce staggering upfront costs. We investigated the correlation 

between sustained virological response (SVR) and drug 

costs. Methods We derived medication costs from Swiss public 

reimbursement prices and extracted published SVR data of 

currently available HCV therapies (ribavirin, pegylated interferon, 

simeprevir, boceprevir, telaprevir, sofosbuvir, ledipasvir, 

ombitasvir, paritaprevir, dasabuvir). We stratified cost-analysis 

by HCV genotype and presence/absence of cirrhosis (in treatment 

naïve patients). We measured the correlation between 

the costs and SVR. All cost results are reported in USD or CHF, 

as with the rounded exchange rate of 1 USD=1 Swiss franc 

might be considered (2015). Results For both patient groups, 

we found a correlation between SVR and cost, where R 2 was 

higher for naive cirrhotic patients (67.8%) compared to naive 

the non-cirrhotic patient group (56.4%) indicating that the 

more one spends on a treatment strategy, the higher the SVR. 

The costs extended between USD12,690 and USD133,750 

per SVR (Fig.1). Discussion There is a significant correlation 

between the rate of SVR and the costs of treatment strategy. 

Payment per patient recovery, regardless of dose and treatment 

duration, including risk-sharing with drug manufacturers 

would allow better budget control and long-term projections. 

Alternatively, pricing volume per year would also minimize 

resources consumption and maximize health benefits and allow 

the manufacturer to provide large amount of drug before the 

era of DAA generics.


(IQR 24-52 months) with negative HBV-DNA. HCC incidence 

was higher in cirrhotic patients (global – 15.6%; 3.8% per 

year) than in patients without cirrhosis (global 1%; 0.2% per 

year), p


intrahepatic virological and inflammatory parameters (Median 

of HBV tDNA: 7.90 vs. 7.45Log 10 

copies/10 6 cells, P = 0.593; 

Median of cccDNA: 6.1 vs. 5.0Log 10 

copies/10 6 cells, P = 

0.052; Percentage of inflammation ≥ G2: 77.8% (21/27) 

vs. 84.6% (11/13), P = 0.933). Furthermore, comparable 

levels of intrahepatic HBV tDNA (Median: 7.89 vs. 7.90Log- 

10 copies/106 cells, P = 0.395) and significantly lower cccDNA 

(Median: 4.75 vs. 6.82Log 10 

copies/10 6 cells, P = 0.003) were 

presented in the patients with undetectable serum HBV DNA 

and normal ALT compared to the patients wits undetectable 

serum HBV DNA and abnormal ALT, and 61.5% (8/13) of the 

patients with undetectable serum HBV DNA and normal ALT 

were found intrahepatic inflammation ≥ G2. Conclusion: More 

than 60% HBV-related compensated cirrhosis patients with 

serum HBV DNA undetectable or at low levels and normal ALT 

had active intrahepatic inflammation and HBV replication, thus 

the administration of antiviral therapy might be strengthened. 

Disclosures: 

The following authors have nothing to disclose: Taotao Yan, Xiaoying Sha, Jing 

Wang, Li Jin, Yu Zhang, Ruitian Yi, Furong Cao, Yuan Yang, Jinfeng Liu, Yingli 

He, Tianyan Chen, Yingren Zhao 

1542 

Comparing antiviral efficacy of Tenofovir monotherapy 

and Tenofovir based combination therapy in antiviral 

resistant chronic hepatitis B: A interim result of multicenter 

cohort study 

Kyu sik Chung 1 , Beom Kyung Kim 1 , Ki Tae Yoon 2 , Hana Park 3 , 

Chang Wook Kim 4 , Jin-Woo Lee 5 , Young Seok Kim 6 , Jung Il Lee 7 , 

Jun Yong Park 1 , Seung Up Kim 1 , Do Young Kim 1 , Sang Gyune 

Kim 6 , Young-Joo Jin 5 , Hee Yeon Kim 4 , Seong Gyu Hwang 3 , Mong 

Cho 2 , Kwan Sik Lee 7 , Kwang-Hyub Han 1 , Sang Hoon Ahn 1 ; 

1 Department of Internal Medicine, Yonsei University College of 

Medicine, Seoul, Korea (the Republic of); 2 Department of Internal 

Medicine, Pusan National University School of Medicine, Yangsan, 

Korea (the Republic of); 3 Department of Internal Medicine, 

CHA Bundang Medical Center, Bundang, Korea (the Republic of); 

4 Department of Internal Medicine, The Catholic University of Korea 


of Internal Medicine, Inha University College of Medicine, Incheon, 

Korea (the Republic of); 6 Department of Internal Medicine, Soonchunhyang 

University College of Medicine, Cheonan, Korea (the 

Republic of); 7 Department of Internal Medicine, Gangnam Severance 

Hospital, Seoul, Korea (the Republic of) 

Background: We aimed to compare antiviral efficacy between 

tenofovir (TDF) monotherapy and TDF-based combination therapy 

as a rescue therapy for chronic hepatitis B with resistance 

to antiviral agents. Methods: A total of 847 consecutive CHB 

patients treated with TDF monotherapy or TDF-based combination 

therapy (TDF plus entecavir, lamivudine or telbivudine) 

as a rescue therapy for resistance to antiviral agents were 

analyzed. Complete virological response (CVR) and biochemical 

response were defined as undetectable serum HBV-DNA 

(detection limit 20 IU/mL) and normalization of serum alanine 

aminotransferase (ALT) level, respectively. Results: At the time 

of rescue therapy, the median serum HBV DNA and ALT level 

were 3.36 log IU/mL and 28 IU/mL, respectively. During the 

follow-up (median 18.3 month), cumulative rates of CVR at 

24 months among patients with resistance to lamivudine only 

(n=474) were similar between those with TDF monotherapy 

and those with TDF-based combination therapy (95.6% vs. 

92.4%, respectively). Similar results (95.5% vs. 94.0%, respectively) 

were maintained in patients with multidrug-resistance 

(n=373) (resistance to lamivudine and entecavir [n=189], resistance 

to lamivudine and adefovir [n=137], and resistance to 

lamivudine, adefovir, and entecavir [n=47]) (all p>0.05). In 

HBeAg- positive patients, cumulative HBeAg seroconversion 

rate at 24 months was comparable between those with TDF 

monotherapy and those with TDF-based combination therapy 

(7.6% vs. 11.8%, P=0.258). Conclusions: TDF monotherapy 

showed similar antiviral efficacy compared with TDF-based 

combination therapy as a rescue therapy, even in patients with 

multi-drug resistance. Further studies with a longer follow-up 

are required to validate these results 

Disclosures: 

Ki Tae Yoon - Grant/Research Support: Handok; Speaking and Teaching: Gilead, 

BMS, MSD, Roche, GSK 

Chang Wook Kim - Consulting: Gilead Korea, MSD; Grant/Research Support: 

BMS Korea, Daewoong, Handok, Pharmicell, Pharmaking, Gilead Korea; Speaking 

and Teaching: BMS Korea, Daewoong 

The following authors have nothing to disclose: Kyu sik Chung, Beom Kyung Kim, 

Hana Park, Jin-Woo Lee, Young Seok Kim, Jung Il Lee, Jun Yong Park, Seung Up 

Kim, Do Young Kim, Sang Gyune Kim, Young-Joo Jin, Hee Yeon Kim, Seong Gyu 

Hwang, Mong Cho, Kwan Sik Lee, Kwang-Hyub Han, Sang Hoon Ahn 

1543 

Hepatitis B virus infection in irregular and refugee 

migrants in Naples, Italy 

Nicola Coppola 1 , Loredana Alessio 1 , Luciano Gualdieri 2 , Mariantonietta 

Pisaturo 3 , Caterina Sagnelli 4 , Nunzio Caprio 5 , Carmine 

Minichini 1 , Mario Starace 1 , Giuseppe Signoriello 1 , Giuseppe 

Pasquale 1 , Evangelista Sagnelli 1 ; 1 Mental Health and Public Medicine, 

Second University of Naples, Naples, Italy; 2 Medical Center, 

Centro per la Tutela della Salute degli Immigrati, Naples, Italy; 

3 Medical Center, Centro di Accoglienza “La tenda di Abramo”, 

Caserta, Italy; 4 Medical Center, Centro Sociale ex Canapificio, 

Caserta, Italy; 5 Medical center, Centro Suore Missionarie della 

Carità, Naples, Italy 

Aim: to define the characteristic of HBV infection in a cohort of 

irregular or refugee immigrants living in Italy, a country with an 

HBsAg prevalence nearly 1% and a HBV universal vaccination 

covering subjects under 35. Methods: A screening for HBV, 

HCV and HIV infections was offered free of charge and of 

bureaucratic procedures to 1,254 illegal or refugee immigrants 

living from 5 years or more in Naples or in its surroundings 

(Italy). Of these 1,254, 1,212 (96.6%), 831 irregular and 

381 refugees, accepted to be screened at one of 4 first-level 

clinical centers operating for years in this setting, with proven 

experience in clinical, psychological and legal management 

of vulnerable groups. The median age of screened subjects 

was 32 years, range 12-74, 75.2% were males, 52% came 

from Sub-Saharan Africa, 18% from Eastern-Europe, 13% from 

Indo-Pakistan Area, 7% from Northern-Africa and the remaining 

10% from others countries. Positive subjects ware further 

investigated at two third-level liver units. Results: One-hundred 

sixteen migrants (9.6%) were HBsAg positive, 490 (40.4%) 

HBsAg negative/anti-HBc positive and 606 (50%) sero-negative 

for both. All positive subjects ignored their serological 

status. A high HBsAg sero-prevalence was found in migrants 

from sub-saharan Africa (13.8%) and an intermediate one in 

those from Eastern Europe (6%), Northern Africa (3.4%) and 

Indo-Pakistan area (2.8%). A logistic regression analysis identified 

as factors independently associated with HBV infection 

(both HBsAg positive and HBsAg negative/anti-HBc positive) 

the male gender (p=0.002), the sub-Saharan African origin 

(p


of infectious diseases: only 4 (5.1%) were HBeAg positive and 

2 (2.5%) anti-Delta positive. According the EASL criteria, 58 

(73.4%) were considered asymptomatic carriers, 17 (21.5%) 

with chronic hepatitis, 2 with liver cirrhosis and 2 with liver 

cirrhosis plus HCC. Conclusions: The HBsAg sero-prevalence is 

very high in the immigrants investigated, particularly in those 

from Sub-Saharan Africa who are introducing HBV-genotype E 

in Italy. The data suggest a stronger action of the Healthcare 

Authorities to limit the diffusion of HBV infection in Italy by 

supporting screenings, educational programs, HBV vaccination 

and treatments in favor of migrant populations. 

Disclosures: 

The following authors have nothing to disclose: Nicola Coppola, Loredana 

Alessio, Luciano Gualdieri, Mariantonietta Pisaturo, Caterina Sagnelli, Nunzio 

Caprio, Carmine Minichini, Mario Starace, Giuseppe Signoriello, Giuseppe 

Pasquale, Evangelista Sagnelli 

1544 

Four years real-life experience with antiviral adherence 

in chronic hepatitis B infection in a tertiary university 

hospital 

Rodrigo M. Abreu 1,2 , Leda C. Bassit 3 , Sijia Tao 3 , Yong Jiang 3 , 

Denise Paranaguá-Vezozzo 1 , Raymond F. Schinazi 3 , Flair J. Carrilho 

1 , Suzane K. Ono 1 ; 1 Gastroenterology, University of São 

Paulo School of Medicine, São Paulo, Brazil; 2 Division of Pharmacy, 

Clinical Hospital, University of São Paulo School of Medicine, 

São Paulo, Brazil; 3 Pediatrics, Emory University School of 

Medicine, and Veterans Affairs Medical Center, São Paulo, Brazil 

BACKGROUND: Evidence shows that antiviral treatment for 

chronic hepatitis B (CHB) infection suppress viral load, but does 

not eradicate hepatitis B virus (HBV). Among the factors directly 

linked to therapeutic success is adherence to treatment. The purpose 

of this study was to evaluate the causes of non-response 

to antiviral treatment in chronic HBV infection. METHODS: 

A prospective cohort study with CHB treated with adefovir, 

entecavir, lamivudine and / or tenofovir was performed in 

a Brazilian reference tertiary hospital. Treatment adherence 

was evaluated by a validate questionnaire named CEAT-HBV 

within December 2010 and August 2011. HBV drug resistance 

mutation and single-dose pharmacokinetics were determined 

by PCR sequencing and LC-MS respectively. The IRB approved 

the research study. RESULTS: Of the 183 individuals enrolled, 

CEAT-HBV identified 104 (57%) on HBV treatment adherence. 

Among the 79/183 (43%) individuals with non-adherence 

to antiviral treatment, 53/79 (67.1%) were more frequently 

viral load positive. The frequencies of HBV genotypes were A 

(14%), A1 (34%), A2 (3%), C (18%) and D (14%). However, 

38% (70/183) had positive viral loads suggesting non-response 

to antiviral treatment. In the adherence group (n = 14) 

with positive viral load, 9 (64%) individuals had HBV with drug 

resistance mutations. In the positive viral load and non-adherence 

group (n = 40), 33 (83%) individuals had drug resistance 

HBV mutations. Sixteen out of 70 HBV positive plasma samples 

could not be amplified. The main factors involved with 

non-response to antiviral treatment were drug resistance mutations 

in HBV (51%), non-adherence without drug resistance 

mutations in HBV (37%), short treatment duration (8%) and 

non-determined (4%). The single-dose pharmacokinetics was 

used to confirm antiviral non-adherence. Two years after the 

first evaluation, the CEAT-HBV showed that 106/143 (74.1%) 

of individuals were on treatment adherent. However, 21.2% 

(40/183) individuals could not be evaluated and excluded. 

The main reasons for exclusion were death (20/183), 10 out 

20 deaths due to hepatocellular carcinoma, loss to follow up 

(11/183) and others (9/183). It is worth noting that these 10 

dead persons were adherent to antiviral treatment and had 

undetectable HBV DNA. Conclusion: Non-adherence was high 

among CHB individuals receiving antiviral treatment. Drug 

resistance mutations and non-adherence to anti-HBV therapy 

were the main cause of treatment failure in CHB individuals. 

This study demonstrates the urgency to implement more effective 

procedures to improve adherence to antiviral treatment for 

chronic hepatitis B. 

Disclosures: 

Raymond F. Schinazi - Board Membership: Cocrystal Pharma, Inc.; Stock Shareholder: 

Cocrystal Pharma, Inc. 

The following authors have nothing to disclose: Rodrigo M. Abreu, Leda C. Bassit, 

Sijia Tao, Yong Jiang, Denise Paranaguá-Vezozzo, Flair J. Carrilho, Suzane 

K. Ono 

1545 

Factors Associated with the Development of Hepatocellular 

Carcinoma after HBeAg Seroclearance 

James Fung, Ching-Lung Lai, Danny Wong, Wai-Kay Seto, Man- 

Fung Yuen; University of Hong Kong, Hong Kong, China 

Background: The role of core promoter mutation, precore mutation, 

and genotype in the development of hepatocellular carcinoma 

(HCC) in patients after HBeAg seroclearance is unclear. 

Methods: The CHB E-Seroclearance Study (C.H.E.S.S.) cohort 

follows up a large population of patients with documented 

HBeAg seroclearance. Surveillance for HCC was performed at 

regular intervals with alpha-fetoprotein and ultrasonography. 

This substudy includes patients who had precore and core promoter 

mutation and genotype determined at 3 to 24 months 

after the time of HBeAg seroclearance. Results: A total of 293 

CHB subjects were included with a median follow up of 124 

months (range, 15 to 254) after HBeAg seroclearance, totaling 

3,200 patient-years. The median age of HBeAg seroclearance 

was 34 years, of which 164 (56%) were male. The presence 

of core promoter and precore mutation was observed in 181 

(61.8%) and 249 (85.0%) patients respectively. The majority of 

patients were of genotype C (88.1%). The cumulative rates of 

HCC at 5, 10 and 20 years after HBeAg seroclearance were 

3.5%, 6.9%, and 8.8% respectively. There were a total of 19 

cases of HCC, with a median time to HCC development (from 

the time of HBeAg seroclearance) of 57 months (range, 17 

to 192). The median age of HCC development was 50 years 

(range, 33 to 72). Males had a significantly higher rate of HCC 

compared to females (12.5% vs 3.8% at 20 years respectively, 

p=0.037). The median age of HBeAg seroclearance was significantly 

higher for those who subsequently developed HCC 

compared to those without HCC (43 vs 34 years respectively, 

p


Wai-Kay Seto - Advisory Committees or Review Panels: Gilead Science; Speaking 

and Teaching: Bristol-Myers Squibb 








The following authors have nothing to disclose: James Fung, Danny Wong 

1546 

Assessing the impact of hepatitis B immune globulin 

(HBIG) on responses to the hepatitis B vaccine during 

co-administration 

Qingwen Cui 1 , Iryna Zubkova 1 , Trudy Murphy 2 , Sarah F. Schillie 2 , 

Marian E. Major 1 ; 1 CBER/FDA, Alexandria, VA; 2 Division of Viral 

Hepatitis, Centers for Disease Control and Prevention, Atlanta, GA 

Background: Administration of hepatitis B vaccine (HepB) and 

hepatitis B immune globulin (HBIG) is recommended for infants 

born to hepatitis B virus (HBV) carrier mothers and for exposed 

persons with no known HBV protection. The recommended prophylaxis 

for infants typically consists of HepB and HBIG at birth 

followed by vaccination at 1 and 6 months; recommendations 

specify administering HepB and HBIG via the intramuscular 

(i/m) route at different sites. However, there are documented 

instances of HepB and HBIG being administered at the same 

site (i.e., in the same limb) and the impact of this on immune 

responses to the vaccine remains unanswered. Our goal was 

to address this issue through a series of studies immunizing 

mice with a licensed HepB (GSK Engerix-B) alone or in combination 

with HBIG (Nabi-HB, Biotest Pharmaceuticals). BALB/c 

mice have been routinely used for in vivo potency assays for 

HepB and provide a viable model to study responses. Methods: 

Six week old mice (15/group) were immunized i/m with 

HepB and/or HBIG at a 1:1 v/v ratio at 0, 4, and 16 weeks. 

Groups consisted of: 1) HepB alone 2) HepB in one leg, HBIG 

in a different leg (HepB boost at 4 and 16 wks) 3) HepB and 

HBIG at the same site in the same leg (HepB boost at 4 and 16 

wks) 4) HepB and HBIG mixed prior to administration (O/N 

+4C) (HepB boost at 4 and 16 wks) 5) HBIG alone (no HepB 

boost) 6) Untreated. Animals were bled at 2, 6 and 26 weeks 

and sera were assessed for anti-HBs titers using a commercial 

assay. Results: At week 26 we observed 100% seroconversion 

(>10mIU/mL) in all groups that received HepB, regardless 

of how HepB was administered with HBIG. However, we 

observed significantly lower anti-HBs titers at 26 weeks when 

HBIG and HepB were administered in the same limb (Gp3) 

or after incubation overnight (Gp4) compared to HepB alone 

(Gp1) (p=0.01 and p=0.02, respectively). At week 2 post 

inoculation, prior to the development of responses to vaccine, 

circulating levels of HBIG were lower in Gp3 mice (p=0.07; 

87% seropositive) and Gp4 mice (p=0.001; 66% seropositive) 

that received HepB and HBIG in the same limb compared to 

those that received HBIG and HepB in different limbs (Gp2). 

We observed similar results following inoculation of 3 week 

old mice to simulate neonates. Conclusions: Administration of 

HepB and HBIG in the same limb does not impact the long-term 

response to HepB in either adult or young mice. However, 

this incorrect method of administration can impact the initial 

levels of circulating HBIG prior to the development of adaptive 

immune responses which in some individuals could result in no 

protective antibody levels during the first weeks after exposure 

to HBV. 

Disclosures: 

The following authors have nothing to disclose: Qingwen Cui, Iryna Zubkova, 

Trudy Murphy, Sarah F. Schillie, Marian E. Major 

1547 

End-of-therapy Serum Gradient of Hepatitis B Surface 

Antigen Predicts Off-therapy Durability after Discontinuation 

of Nucleos(t)ide Analogues 

Yao-Chun Hsu 1,2 , Chun-Ying Wu 3 , Chi-Yang Chang 1 , Ming-Shiang 

Wu 7 , Jia-Horng Kao 4 , Tzeng-Huey Yang 5 , Hsu-Wei Hung 8 , 

Jaw-Town Lin 6 ; 1 Division of Gastroenterology, E-Da Hospital, Kaohsiung, 

Taiwan; 2 School of Medicine, I-Shou University, Kaohsiung 

City, Taiwan; 3 Division of Gastroenterology, Taichung Veterans 

General Hospital, Taichung City, Taiwan; 4 Graduate Institute of 

Clinical Medicine, National Taiwan University, Taipei City, Taiwan; 

5 Division of Gastroenterology, Lotung Poh-Ai Hospital, Yilan 

County, Taiwan; 6 School of Medicine, Fu Jen Catholic University, 

New Taipei City, Taiwan; 7 Department of Internal Medicine, 

National Taiwan University, Taipei City, Taiwan; 8 Taipei Institute 

of Pathology, Taipei City, Taiwan 

Background and Aims: Relapse of chronic hepatitis B (CHB) 

is common but remains unpredictable after discontinuation of 

nucleos(t)ide analogues (NAs). This multicenter prospective 

research aimed to explore the association between serum gradient 

of hepatitis B surface antigen (HBsAg) and off-therapy 

durability following NA cessation. Methods: From July 2011 

through December 2014, 228 consecutive CHB patients who 

were about to discontinue NAs were screened from 3 teaching 

hospitals in Taiwan. Those who had achieved undetectable 

viral DNA after a minimum of 3-year therapy were included. 

Participants were monitored until February 2015 for occurrence 

of clinical relapse, stringently defined as viral DNA >2,000IU/ 

mL plus ALT >2 folds upper normal limit for ^3 months, and 

viral relapse, simply defined as viral DNA>2,000 IU/mL. Incidences 

of relapse were evaluated by the Kaplan Meier method 

and risk predictors determined by the Cox proportional hazard 

analysis. Results: Among 158 eligible patients observed for a 

mean duration of 14.6 (range, 3-42.2) months, clinical and 

viral relapses occurred in 44 and 103 patients, respectively, 

corresponding to 2-year cumulative incidences of 46.2% (95% 

CI, 35.2-58.8%) and 81.6% (95% CI, 72.9-88.8%), respectively. 

In 119 patients with negative HBeAg at NA cessation, 

end-of-therapy HBsAg was associated with both clinical and 

viral relapse in a dose-response manner (Ptrend=0.02 for clinical 

and 0.0005 for virological relapse). Patients whose serum 

HBsAg fell below 100 IU/mL did not develop clinical relapse 

but could experience viral relapse with a cumulative incidence 

of 33.4% (95% CI, 14.6-64.7%) at 2 years. In those with 

HBsAg


Disclosures: 

Yao-Chun Hsu - Speaking and Teaching: GlaxoSmithKline, Novartis, Bristol-Myers 

Squibb Company, Harvester Trading Company 

The following authors have nothing to disclose: Chun-Ying Wu, Chi-Yang Chang, 

Ming-Shiang Wu, Jia-Horng Kao, Tzeng-Huey Yang, Hsu-Wei Hung, Jaw-Town 

Lin 

1548 

Liver fibrosis progression, new-onset metabolic syndrome 

and risk of hepatocellular carcinoma in chronic 

hepatitis B – an 8-year prospective cohort study of 

1,178 patients with paired transient elastography 

examination 

Grace LH Wong, Henry Lik-Yuen Chan, Angel ML Chim, Vincent 

W. Wong; Institute of Digestive Disease, The Chinese University of 

Hong Kong, Shatin, Hong Kong 

Background: New-onset metabolic syndrome (nMES) was 

recently found to increase the risk of liver fibrosis progression 

in patients with chronic hepatitis B (CHB). We aimed to investigate 

the effect of nMES on the risk of hepatocellular carcinoma 

(HCC) in CHB patients. Methods: 1,466 CHB patients 

underwent liver stiffness measurement (LSM) by transient elastography 

in 2006-2008; 1,178 patients had 2 nd LSM in 2010- 

2012. Liver fibrosis progression was defined as an increase in 

LSM ≥30% at the second assessment. Patients were prospectively 

followed up with regular ultrasonographic examinations 

and 6-monthly alpha-fetoprotein for HCC surveillance since the 

first LSM. The impact of liver fibrosis progression and nMES 

on HCC development was evaluated. Results: At 1 st LSM, the 

mean age of these 1,178 patients was 46±11 years, 63% 

were males, body-mass index (BMI) 23.2±3.3kg/m 2 , serum 

alanine aminotransferase (ALT) 63±40IU/l, 26% HBeAg positive, 

HBV DNA 4.5±2.0logIU/ml, and LSM 8.0±3.6kPa. At 

2 nd LSM, the mean BMI was 23.2±3.5kg/m 2 , ALT 38±35IU/l, 

HBV DNA 2.5±1.8logIU/ml, and LSM was 6.2±3.7kPa; 44% 

had received antiviral treatments. Metabolic syndrome was 

diagnosed in 153 (13%) and 350 (30%) patients at 1 st and 

2 nd LSM; 230 (20%) and 33 (3%) patients had nMES and 

resolved MES respectively. After an interval of 44±7 months, 

155 (13%) patients developed liver fibrosis progression. At 

93±9 months from 1 st LSM, 28 patients (2.4%) developed HCC 

and 5 patients (0.4%) died. Liver fibrosis progression but not 

nMES was the independent risk factor of HCC. The risk of HCC 

in patients with or without liver fibrosis progression at 3 years 

after 2 nd LSM was 4.5% (95% CI 1.1%-9.9%) and 1.6% (95% 

CI 1.0%-2.2%) respectively (P=0.008). The 3-year risk of HCC 

in patients with nMES, compared to those with no MES at both 

time points, was 1.3% (95% CI 0.1%-2.5%) vs. 2.3% (95% CI 

1.3%-3.3%) respectively (P=0.28). Conclusions: Liver fibrosis 

progression but not nMES increases the risk of HCC in CHB 

patients. 

Cumulative probability of HCC in patients with or without liver 

fibrosis progression and new-onset MES. 

Disclosures: 

Grace LH Wong - Advisory Committees or Review Panels: Otsuka, Gilead; 

Speaking and Teaching: Echosens, Furui, Gilead, Janssen, Bristol-Myers Squibb, 

Otsuka, Abbvie 




Vincent W. Wong - Advisory Committees or Review Panels: AbbVie, Gilead, 

Janssen; Consulting: Merck, NovaMedica; Speaking and Teaching: Gilead, 

Echosens 

The following authors have nothing to disclose: Angel ML Chim 

1549 

High Rates of Surface Antigen Loss and Low Rates of 

Seroreversion in Asian Chronic Hepatitis B Patients 

Treated with Oral Antiviral Therapy: Community-Based 

Real World Outcomes 

Robert J. Wong 1 , My T. Nguyen 2 , Huy N. Trinh 3 , Andrew Huynh 2 , 

Mytop Ly 2 , Huy Nguyen 3 , Khanh Nguyen 3 , Jenny D. Yang 3 , Ruel 

Garcia 3 , Brian S. Levitt 3 , Eduardo DaSilvera 3 , Robert Gish 4 ; 1 Gastroenterology 

and Hepatology, Alameda Health System - Highland 

Hospital, Oakland, CA; 2 Silicon Valley Research Institute, San 

Jose, CA; 3 San Jose Gastroenterology, San Jose, CA; 4 Gastroenterology 

and Hepatology, Stanford University Medical Center, 

Stanford, CA 

Background:Chronic hepatitis B virus (HBV) infection is a global 

epidemic with over 240 million persons chronically infected 

with the majority from Asian-Pacific regions. The most definitive 

endpoint of antiviral therapy, surface antigen (HBsAg) loss, is 

rare. It is not clear if HBsAg loss correlates well with baseline 

patient characteristics, undetectable virus and normalization 

of alanine aminotrasferase (ALT) on treatment, or risk of seroreversion 

or detectable virus after stopping therapy. Aim:To 

evaluate rates of HBsAg loss, baseline correlates of HBsAg 

loss, normalization of ALT and undetectable virus, and rates 

of HBsAg seroreversion or re-emergence of detectable virus 

among chronic HBV patients. Methods:A retrospective cohort 

study evaluated 949 adults (age >18) with chronic HBV on 

oral antiviral therapy at a large community gastroenterology 

clinic from June 1997 to May 2015. The majority (99.6%) 

were Asian ethnicity. Rates of HBsAg loss, normalization of 

ALT, achieving undetectable virus, and developing surface 

antibody (anti-HBs) were stratified by HBeAg status. Following 

HBsAg loss, rates of HBsAg seroreversion or re-emergence of 

detectable virus were analyzed. Multivariate logistic regression 

models evaluated for baseline and on-treatment predictors of 

HBsAg loss. Results:With a mean follow up of 86.6 months 

and mean duration of treatment of 81.5 months, overall rate 

of HBsAg loss was 4.74% (n=45), with similar rates between 

HBeAg positive and negative patients (Table). Among HBsAg 

loss patients, 30.9% developed anti-HBs, 91.1% achieved 

undetectable virus, and 64.4% normalized ALT. Only 2.22% 

(n=1) who achieved HBsAg loss had detectable virus, whereas 

13.3% (n=6) of HBsAg loss patients had seroreversion of positive 

HBsAg within 1-17 months. All 6 patients who seroreverted 

had sustained undetectable virus. While there was a 

trend towards higher odds of HBsAg loss among men (OR 

2.05, 95% CI 0.92-4.59, p=0.08), no significant associations 

between baseline characteristics (age, ALT, platelets, HBeAg, 

FIB-4 score, APRI score) and HBsAg loss were seen. Conclusion:Among 

a large community-based gastroenterology clinic, 

treatment of chronic HBV achieved high rates of HBsAg loss 

among Asian patients. While 13.3% of patients experienced 

seroreversion, all of these patients remained viral load undetectable, 

demonstrating the sustainability of HBsAg loss in most 

patients and stable suppression of HBV viral load.


Disclosures: 







The following authors have nothing to disclose: Robert J. Wong, My T. Nguyen, 

Andrew Huynh, Mytop Ly, Huy Nguyen, Khanh Nguyen, Jenny D. Yang, Ruel 

Garcia, Brian S. Levitt, Eduardo DaSilvera 

1550 

The role of hepatitis B core-related antigen in predicting 

HBV reactivation among HBsAg-negative, anti-HBc-positive 

individuals undergoing immunosuppressive therapy 

Wai-Kay Seto, Danny Wong, Sze Hang Kevin Liu, James Fung, 

Ching-Lung Lai, Man-Fung Yuen; Medicine, The University of Hong 

Kong, Queen Mary Hospital, Hong Kong, Hong Kong 

Background: Hepatitis B core-related antigen (HBcrAg) detects 

an identical amino-acid sequence shared by hepatitis B e 

antigen, hepatitis B core antigen and 22kDa precore protein. 

Serum HBcrAg correlates with disease activity of chronic hepatitis 

B (CHB), and can be detected in a considerable proportion 

of CHB patients after hepatitis B surface antigen (HBsAg) 

seroclearance. Its role in predicting HBV reactivation among 

HBsAg-negative, antibody to hepatitis B core antigen (anti-HBc) 

positive has not been explored. Methods: We retrieved stored 

plasma samples of HBsAg-negative, anti-HBc positive, patients 

with undetectable HBV DNA at baseline who were enrolled into 

two prospective studies, which aimed at investigating the rate 

of HBV reactivation during rituximab-containing chemotherapy 

and hematopoietic stem-cell transplantation (HSCT) (ClinicalTrials.gov 

identifier NCT01502397 and NCT01481647 

respectively). HBV reactivation was defined as detectable HBV 

DNA (≥10 IU/mL). Serum HBcrAg (lower limit of detection 

100 U/mL) was measured using a chemiluminscent enzyme 

immunoassay (Fujirebio Inc, Tokyo, Japan) at baseline and 

at every 3 months up to the date of last follow-up. Results: 

We included 131 HBsAg-negative, anti-HBc positive patients 

(rituximab/HSCT 47.3%/52.7%), with a mean age of 57.4 

(±15.1) years and a mean follow-up duration of 53.3 (±35.0) 

weeks. 32 patients developed HBV reactivation (rituximab/ 

HSCT 56.3%/43.7%); the 2-year cumulative rate of HBV 

reactivation was 39.2%. 21 patients (16.0%) and 25 patients 

(19.1%) had detectable serum HBcrAg at either baseline or 

any time point before reactivation respectively. The median 

detectable baseline HBcrAg level was 400 (range 200-5,300) 

U/mL. Baseline serum HBcrAg positivity, compared to baseline 

HBcrAg-negative patients, had a significantly higher 2-year 

cumulative rate of HBV reactivation (75.8% versus 29.9% 

respectively, p


come of HBsAg loss/seroconversion, with a PPV of 83%. NCP 

on treatment was associated with a NPV of 69%. Discussion 

The measurement of the loss or gain of epitope recognition on 

TDF therapy revealed selection pressures directed against the 

HBsAg. Detection of the HBsAg CP by epitope mapping may 

be a potential viral biomarker, possibly reflecting emerging 

anti-HBs selection pressure. These findings may help individualising 

patient therapy, though further validation in expanded 

datasets and with other HBV genotypes is needed. 

Disclosures: 

Julianne Bayliss - Grant/Research Support: Gilead Inc. 

Peter A. Revill - Grant/Research Support: Gilead Sciences 

Thomas Leary - Employment: Abbott Laboratories 

Anuj Gaggar - Employment: Gilead Sciences, Inc. 


Kathryn M. Kitrinos - Employment: Gilead Sciences; Stock Shareholder: Gilead 

Sciences 




Abbvie, BMS 


Health 

The following authors have nothing to disclose: Renae Walsh, Rachel Hammond, 

Lilly Yuen, Howard Thomas 

1552 

Comparison between spontaneous and post-nucleos(t) 

ide analogue HBsAg seroclearance in chronic HBV 

infection 

Yi-Cheng Chen, Rachel Wen-Juei Jeng, Rong-Nan Chien, Chia- 

Ming Chu, Yun -Fan Liaw; Liver Research Unit, Chang Gung 

Memorial Hospital, Taoyuan, Taiwan 

Background and Aims The prognosis following spontaneous 

HBsAg seroclearance in patients with chronic hepatitis B is 

excellent, except in those with cirrhosis or concurrent hepatitis 

C or D virus (HCV or HDV) infection. HBsAg seroclearance 

after nucleos(t)ide analogue (NUC) treatment is associated 

with favorable clinical outcomes. Whether there is difference 

between spontaneous and post-NUC HBsAg seroclearance is 

unknown. Methods Eight-eight patients with HBsAg seroclearance 

after NUC treatment (group A) and 88 age (±1 year) 

and gender-matched patients with spontaneous HBsAg seroclearance 

(group B) were included for analysis. Patients with 

coinfection of HCV or HDV, exposure to interferon and organ 

transplantation have been excluded. HBsAg seroclearance 

was defined as persistent absence of HBsAg for at least 12 

months and until last visit. Results The median age at HBsAg 

seroclearance was 49 years and 78% were males. The proportion 

of parenchymal fatty change at HBsAg seroclearance was 

comparable in both groups (p=1.000). The rate of cirrhosis 

when HBsAg seroclearance occurred was significantly higher 

in group A (21.6% vs. 4.5%, p=0.002). During a median 

follow-up period of 67.8 months in group A and 82.2 months 

in group B, one hepatocellular carcinoma (HCC) developed in 

each group with an estimated annual incidence of 0.2% and 

0.16%, respectively (p=1.000). The cumulative incidence of 

HBsAg seroconversion at 1 year, 3 years and 5 years after 

HBsAg seroclearance was 37.6% and 22%, 60.2% and 

39.9%, 67.1% and 53.5% in group A and group B, respectively 

(p=0.003 at 5 years). Conclusions There was significant 

higher rate of cirrhosis in patients with NUC treatment when 

HBsAg seroclearance occurred. Even in low incidence, HCC 

could develop in patients after HBsAg seroclearance. Long-term 

follow-up is needed to elucidate this untoward event. 

Disclosures: 

Yun -Fan Liaw - Advisory Committees or Review Panels: Roche; Grant/Research 


The following authors have nothing to disclose: Yi-Cheng Chen, Rachel Wen-Juei 

Jeng, Rong-Nan Chien, Chia-Ming Chu 

1553 

Different DNA methylation patterns by viral status in 

liver cancer 

Min-Ae Song 2,4 , Sandi Kwee 3 , Maarit Tiirikainen 2 , Gordon S. 

Okimoto 2 , Brenda Y. Hernandez 2 , Linda L. Wong 2,1 , Naoky CS 

C. Tsai 2 , Herbert Yu 2 ; 1 Surgery, Univ of Hawaii School of Medicine, 

Honolulu, HI; 2 Cancer Center, University of Hawaii, Honolulu, 

HI; 3 PET research, Queens Medical Center, Honolulu, HI; 

4 College of Public Health, Ohio State University, Columbus, OH 

Hepatocellular carcinoma (HCC) is one of the most common 

human cancers with high fatality. There is limited understanding 

of genome-wide differences in DNA methylation by viral 

status in HCC. Methods:We performed genome-wide methylation 

profiling in a total of 33 HCC tumor samples, including 

10 HBV-HCC, 13 HCV-HCC, and 10 non-viral HCC, using the 

Illumina HumanMethylation450 BeadChip. Results:Thirty-nine 

loci were found significantly different in methylation among 

the three groups (p


Disclosures: 

Sandi Kwee - Grant/Research Support: Bayer Healthcare 

Linda L. Wong - Speaking and Teaching: Bayer, Bayer 

Naoky CS C. Tsai - Advisory Committees or Review Panels: Bristol-Myers Squibb, 

Gilead, AbbVie; Consulting: BMS, Gilead; Grant/Research Support: BMS, 

Gilead, AbbVie; Speaking and Teaching: Bristol-Myers Squibb, Gilead, Salix, 

Bayer, AbbVie 

The following authors have nothing to disclose: Min-Ae Song, Maarit Tiirikainen, 

Gordon S. Okimoto, Brenda Y. Hernandez, Herbert Yu 

ng/mL) in this study population, suggesting an opportunity to 

improve the current HCC screening for AFP-negative HCC. To 

validate the urine DNA test for the early detection of primary 

and recurrent HCC, an on-going blinded study was conducted 

from a population of HBV-infected cirrhosis patients, with or 

without previous HCC. All patients were monitored for severe 

liver diseases including HCC. Urine samples were collected 

at visits with a hepatologist and were barcoded and tested 

for three DNA markers. After a course of 6 to 18 months of 

follow-ups, four HCC cases, including three recurrent cases, 

were detected by MRI. More significantly, urine DNA markers 

detected three of four HCC cases at 6 and 9 months prior to 

the time of MRI diagnosis. In conclusion, HCC DNA markers 

can be detected in urine of patients with HCC by short-amplicon, 

PCR-based assays. Furthermore, we have demonstrated 

that in combination with serum AFP, a urine DNA test detected 

at least 30% more HCC in an open-labeled study as compared 

to serum AFP alone, and therefore, this test has the potential 

to detect HCC prior to MRI imaging in a blinded pilot study. 

Su, et. al., J Mol Diag, 6, 101-107. (2004) Lin, et al., J Mol 

Diag 13: 474-484 (2011) Jain, et al., Hepatology Research, 

(2014), doi: 10.1111/hepr.12449 

Disclosures: 

Hie-Won Hann - Advisory Committees or Review Panels: Gilead; Grant/Research 

Support: Gilead, Bristol-Myers Squibb 

Surbhi Jain - Employment: JBS Science, Inc 

Wei Song - Stock Shareholder: JBS Science Inc 

Ying-Hsiu Su - Advisory Committees or Review Panels: JBS Science Inc 

The following authors have nothing to disclose: Selena Lin, Sooji Yi, Ting-Tsung 

Chang, Chi-Tan Hu 

1554 

Detection of genetic and epigenetic DNA markers in 

urine for the early detection of primary and recurrent 


Hie-Won Hann 1 , Surbhi Jain 2 , Selena Lin 6 , Sooji Yi 1 , Ting-Tsung 

Chang 3 , Chi-Tan Hu 4 , Wei Song 2 , Ying-Hsiu Su 5 ; 1 Liver Disease 

Prevention Center, Division of Gastroenterology and Hepatology, 

Thomas Jefferson University Hospital, Philadelphia, PA; 2 JBS Science, 

Inc, Doylestown, PA; 3 National Cheng Kung Universtiy Medical 

College, Tainan, Taiwan; 4 Buddhist Tzu Chi General Hospital 

and Tzu Chi University, Hualien, Taiwan; 5 The Baruch S. Blumberg 

Institute, Doylestown, PA; 6 Drexel University College of Medicine, 


The purpose of this study is to explore the potential of urine 

DNA biomarkers for the early detection of primary and recurrent 

hepatocellular carcinoma (HCC). HCC is an aggressive 

disease with a 5-year survival rate of 26%, in early-stage cancers, 

and a mere 2% in later stages, with an approximate 

50% recurrence rate in the first 2 years of treatment. The most 

commonly used screening biomarker is serum alpha-fetoprotein 

(AFP), which detects only 40-60% of cases. We have 

previously shown that urine contains fragmented, circulation-derived, 

cell-free DNA that can be used for detection of 

cancer-related DNA markers, if the tumor is present (1). In 

order to detect circulation-derived, cell-free DNA markers in 

urine, we have developed short amplicon (~50 bp) PCR-based 

assays for mutations in TP53 (249T) (2) and for aberrant DNA 

methylation in GSTP1 (mGSTP1) and RASSF1A (mRASSF1A 

(3)). In addition, we have shown that the AUROC of three urine 

DNA markers, mGSTP1, mRASSF1A, and TP53 249T mutations, 

in combination with AFP, is 0.98 in detecting primary 

HCC (n=74) from non HCC [(cirrhosis (n=45) and hepatitis 

(n=42)] patients by a logistic regression-based combination 

algorithm. Furthermore, these 3 DNA markers detected 35 of 

the 46 (76%) HCC samples that were AFP negative (< 20 

1555 

Prevalence and serological features of chronic hepatitis 

B patients with concurrent HBsAg and anti-HBs in the 

Hepatitis B Research Network (HBRN). 

William M. Lee 1 , Yona K. Cloonan 2 , Kathleen B. Schwarz 3 , Doan 

Y. Dao 1 , Anna S. Lok 4 ; 1 Division of Digestive and Liver Diseases, 

University of Texas Southwestern Medical Center at Dallas, Dallas, 

TX; 2 Epidemiology, University of Pittsburgh, Pittsburgh, PA; 3 Pediatrics, 

Johns Hopkins University School of Medicine, Baltimore, MD; 

4 Gastroenterology, University of Michigan, Ann Arbor, MI 

Background: Some patients with chronic HBV infection have 

concurrent HBsAg and anti-HBs. We examined the HBRN 

cohort to determine the prevalence of anti-HBs in participants 

with chronic HBV infection, and compared demographic and 

clinical factors between participants with and without anti-HBs. 

Methods: Data from 1402 (1115 adults and 287 pediatric) 

participants with chronic HBV infection enrolled in the HBRN, 

who had been tested for both HBsAg and anti-HBs within 1 

year of enrollment were included. 139 participants belonging 

to specifically targeted groups (e.g., pregnant women) were 

excluded from prevalence estimates. Characteristics of anti-HBs 

positive and anti-HBs negative participants were compared. 

Results: 93 participants were anti-HBs positive, overall prevalence 

of concurrent HBsAg and anti-HBs was 6.6% (7.7% 

pediatric and 6.3% adult). Age, sex, race and birthplace did 

not differ significantly between anti-HBs positive and anti-HBs 

negative participants. Similarly, there were no significant differences 

in mode of transmission, estimated years of infection, 

past HBV treatment, prevalence of HBeAg, HBV genotypes, 

AST, ALT or other biochemical markers of liver injury[SB1] in 

the 2 groups. HBsAg levels were significantly lower in anti-HBs 

positive participants: median 3.0 vs. 3.6 log 10 

IU/mL (p


noted in pediatric and adult groups. Conclusion: Concurrent 

HBsAg and anti-HBs is not uncommon in persons with chronic 

HBV infection. The prevalence of anti-HBs is similar for adults 

and children, and is not associated with markers of disease 

activity or a specific clinical phenotype. Anti-HBs positive participants 

had lower HBsAg titers but similar HBV DNA levels 

suggesting a role for anti-HBs in decreasing sub-virion particles 

or in immune control of HBV infection. Further studies are 

planned to examine surface gene variants and anti-HBs titers in 

this [SB2] population. 

Comparison Data between anti-HBs pos and neg HBsAg positive 

individuals 

Disclosures: 



Kathleen B. Schwarz - Consulting: Novartis; Grant/Research Support: Bristol-Myers 

Squibb, Gilead, Roche/Genentech, Synageva, Vertex, Roche 



The following authors have nothing to disclose: Yona K. Cloonan, Doan Y. Dao 

1556 

Clinical Outcomes of Patients with Private vs. Public 

Drug Coverage for Treatment of Chronic Hepatitis B 

Aman V. Arya, Mayur Brahmania, Matthew Kowgier, Hemant 

Shah, Orlando Cerocchi, David K. Wong, Jordan J. Feld, Harry 

L. Janssen; Gastroenterology, University of Toronto, Toronto, ON, 

Canada 

Background Canada has universal health coverage for basic 

medical care; however, prescription drugs are covered by a 

private insurance (if eligible) or through an exceptional access 

program (EAP) for those without insurance. Coverage for 

chronic hepatitis B treatment (CHB) is restricted by the EAP. 

Aim To compare clinical outcomes between individuals covered 

by EAP and those covered by private insurance for the 

management of CHB. Methods A retrospective chart audit was 

conducted of adult patients with CHB who were on treatment 

with a nucleos(t)ide analogue at a large tertiary care liver clinic 

from April 2010 to March 2015. Rates of HBeAg seroconversion, 

HBsAg loss, time to undetectable HBV DNA and ALT normalization, 

HBV DNA breakthrough, and ALT flares (defined as 

5xULN) on therapy were compared. Results 588 patients (285 

EAP, 303 non-EAP) met study inclusion criteria. There were 409 

males (70%) and the mean age was 52 (range = 19-89). 78% 

were Asian, 15% Caucasian and 4% African. 144 patients 

had cirrhosis (25%), The mean age of EAP patients receiving 

treatment was 56 years compared with 48 for patients with 

private insurance (p < 0.001). EAP patients were less likely 

to be born in Canada (2% vs. 10%, p < 0.001), more likely 

to have cirrhosis (29% vs 20%, p=0.02), and were less likely 

to be employed (49% vs. 77%, p < 0.001). LAM, TDF and 

ETV were used in 64%, 22%, 3% of EAP patients respectively, 

compared to 29%, 39%, 21% in patients with private insurance 

(p < 0.001). HBeAg seroconversion occurred in 42% of 

EAP patients compared to 46% of private insurance patients 

(p=0.89). HBsAg loss occurred in 5% versus 7% (p=0.39) and 

the mean time to HBV DNA negativity was 5 months (range: 

0-46) versus 5 months (range: 0-54) (p=0.17). Time to ALT 

normalization was 3 months and not different between the two 

groups (p=0.55). HBV DNA breakthrough leading to a change 

in therapy was more common among EAP patients (38% vs. 

22%, p


onymous (NS), genotype specific viral substitutions from wild 

type (WT) and entropy scores, were calculated across the 

whole genome (WG) and within specific structural (Envelope; 

Polymerase; precore-core, and; HBx) and regulatory regions 

(negative regulatory element- basal core promoter NRE-BCP) 

and correlated with baseline clinical and virological data and 

serological response at W192. Results 94 patients were successfully 

analysed (median age 32yrs; 75% male; genotype 

A/B/C/D 26%/23%/23%/28%; ALT 139 IU/mL; 24% cirrhosis; 

HBV DNA 8.43 log 10 

IU/mL; HBsAg 50,793 IU/mL; 

HBeAg 1,990 PEIU/mL; 77% TDF therapy W0-W48, 23% 

ADV therapy W0-W48). Median WG coverage, substitutions 

from WT and entropy scores are listed in Table 1. Entropy 

scores were similar across the genome, but three-fold greater 

in genotype B/C infection as compared to A/D (p< 0.0001). 

An increase in WG viral diversity was associated with lower 

baseline HBV DNA, HBeAg and HBsAg titres, independent 

of genotype (p< 0.0001 for all). Higher baseline viral diversity, 

specifically in BCP-NRE, was also associated with a lower 

likelihood of achieving HBeAg SC (p=0.04) and HBsAg loss 

(p


1559 

Genotype-Specific Prevalence and Importance of Naturally 

Occurring Precore-, Basal Core Promoter- and 

preS-Mutations in a Large European Study Cohort of 

Patients Chronically Infected with Hepatitis B Virus 

Lisa Sommer 1 , Kai-Henrik Peiffer 1 , Julia Dietz 1 , Simone Susser 1 , 

Joerg Petersen 2 , Peter Buggisch 2 , Markus Cornberg 3 , Stefan 

Mauss 4 , Hartwig Klinker 5 , Martin F. Sprinzl 6 , Florian van Bömmel 

7 , Eberhard Hildt 8 , Caterina Berkowski 1 , Dany Perner 1 , Sandra 

Passmann 1 , Stefan Zeuzem 1 , Christoph Sarrazin 1 ; 1 Medical 

Clinic 1, J. W. Goethe University Hospital, Frankfurt, Germany; 

2 IFI-Institut an der Asklepiosklinik St. Georg, Hamburg, Germany; 

3 Gastroenterology, Hepatology and Endocrinology, Hannover 

Medical School, Hannover, Germany; 4 Center for HIV and Hepatogastroenterology, 

Düsseldorf, Germany; 5 Internal Medicine II, 

University of Wuerzburg Medical Center, Würzburg, Germany; 

6 I. Medizinische Klinik und Poliklinik, Universitätsmedizin der 

Johannes Gutenberg-Universität Mainz, Mainz, Germany; 7 University 

Hospital Leipzig, Leipzig, Germany; 8 Paul-Ehrlich-Institut, 

Langen, Germany 

Background/Aims In several studies from Asia mutations in 

HBV basal core promoter (BCP) and in the preS region are 

associated with the development of liver fibrosis, cirrhosis and/ 

or hepatocellular carcinoma. For precore (PC) mutations data 

are controversially discussed. In EU/US only limited data about 

the significance of these mutations in chronically infected HBV 

patients (pts.) is available. To establish prognostic markers 

the genotype-specific prevalence of these mutations and their 

importance for progression of disease were examined in a 

large European study cohort of pts. infected with HBV genotypes 

(gt) A to E. Methods In the prospective, longitudinal 

ALBATROS study HBsAg carriers without antiviral therapy will 

be followed for more than 10 years. In the present interim 

analysis baseline (BL) sera of 340 pts. (gtA: 81, gtB: 28, gtC: 

16, gtD: 192, gtE: 23) were examined. Progression of infection 

was controlled up to 5 years follow-up (FU). BCP/PC and 

preS regions were amplified via nested PCR and analyzed 

by population-based sequencing. Results BCP double mutation 

A1762T/G1764A was found frequently in our population 

(203/340; 60%) with the highest prevalence in gtE (91%), 

with moderate frequency (56-69%) in gtA, gtC and gtD and 

with the lowest prevalence in gtB (29%). The PC G1896A 

mutation was detected in 42% (142/340) with the highest 

prevalence in gtB, gtC and gtD (71, 56 and 54%) and the 

lowest prevalence in gtA and gtE (10 and 9%). In addition, the 

PCG1896A/G1899A double mutation was found with moderate 

frequency in gtD, gtB and gtC (27, 18 and 13%) but only 

infrequently in gtA (3%) and was absent in gtE (0%). Deletions 

in the preS region varying in length from 1aa to 41aa occurred 

in 8% of investigated patients with the highest prevalence in 

gtE (40%) and gtC (29%). Seven patients (gtA: 2; gtD: 4; gtE: 

1) started antiviral therapy after 1-4 years of FU. BCP double 

mutation was observed in 6/7 and PC double mutation in 

1/7 patients at BL. Conclusion BCP and PC mutations were 

found frequently in our study cohort in a HBV genotype-specific 

pattern. Deletions in the preS region were found with a lower 

prevalence but were observed predominantly in HBV gtE and 

gtC. Interestingly, 6/7 HBsAg carriers who had to start treatment 

during FU were positive for the BCP double mutation at 

BL, whereas PC mutations were found only infrequently in these 

patients. Therefore, BCP and PC mutations might be of clinical 

interest as potential prognostic markers in HBsAg carriers. 

Disclosures: 













Hartwig Klinker - Advisory Committees or Review Panels: Bristol-Myers Squibb, 

Janssen, Hexal; Grant/Research Support: Gilead, Abbott, AbbVie, Janssen, 

Novartis, Bristol-Myers Squibb; Speaking and Teaching: Gilead, Bristol-Myers 

Squibb, Merck Sharp & Dohme 

Martin F. Sprinzl - Grant/Research Support: GILEAD 




Inc., Abbvie 









The following authors have nothing to disclose: Lisa Sommer, Kai-Henrik Peiffer, 

Julia Dietz, Simone Susser, Eberhard Hildt, Caterina Berkowski, Dany Perner, 

Sandra Passmann 

1560 

Individualizing Hepatitis B Prophylaxis in Liver Transplant 

(LT) Recipients: Diminished Need for Hepatitis B 

Immunoglobulin (HBIG) 

Kavita Radhakrishnan 1 , Aileen Chi 2 , David J. Quan 2 , John P. 

Roberts 3 , Norah Terrault 2 ; 1 Medicine, UCSF, San Francisco, CA; 

2 Transplant Hepatology, UCSF, San Francisco, CA; 3 Transplant 

Surgery, UCSF, San Francisco, CA 

Background & Aims: HBIG has been central to the prevention 

of hepatitis B virus (HBV) recurrence post-LT for two decades. 

However, its high cost and need for IV or IM administration 

have prompted HBIG minimization strategies. Elimination of 

HBIG and use of antiviral therapy (AVT) alone is associated 

with HBsAg recurrence in ~20% at 2 yrs (Fung, Gastroenterology. 

2011). Achieving a higher rate of HBsAg negativity is 

highly desirable. We report our experience with an individualized 

protocol for HBIG prophylaxis. Methods: All HBsAg-positive 

patients undergoing LT from 2009-2014 were included. 

Patients at high risk for recurrence [defined as HBV DNA>100 

IU/mL at time of LT (33%, 2 acute), prior AVT resistance (33%), 

HDV+ (22%), or HIV+ (12%)] received HBIG 5K-10K U in anhepatic 

phase & daily X 5 days, and every 4-12 wks to maintain 

trough anti-HBs >100 U/L. In contrast, low risk patients [all 

others] received HBIG 5K U in anhepatic phase & daily x 5 

days only. Both groups received ATV therapy indefinitely. HBV 

recurrence was defined as HBsAg positivity post-LT. Results: Of 

79 LT recipients, 63% and 37% met criteria for high and low 

risk protocols. Pre-LT characteristics and post-LT prophylaxis 

are shown in Table. Median (IQR) follow-up was 2.9 (1.3-4.4) 

years. The 1 and 3-yr cumulative incidences of HBsAg positivity 

post-LT were 2.0% (CI 95 

: 0.3 -13.4) and 5.4% (CI 95 

: 1.3- 

20.7) in the high risk vs. 0% (upper CI 95 

: 11.9) in the low risk 

groups (p=0.28). Only those who developed metastatic HCC 

became HBsAg-positive (n=2). All patients were HBV DNA 

negative post-LT, except one with recurrent HCC. Post-LT survival 

was 91% and 84% at 3 and 5 years respectively, with no 

difference between prophylaxis groups (p=0.55). Conclusion: 

In patients without HIV and HDV who have HBV DNA levels 


is highly effective in preventing HBV recurrence. With current 

preferred antivirals (high barrier to resistance), more patients 

are likely to have such a profile and utilizing an individualized 

approach to HBIG prophylaxis offers a better balance between 

prophylaxis costs and prevention of HBV recurrence. 

ULN: 575.0 vs 127.9; ALT


ily was detected by Western blot, and the DNA binding level of 

hepatocyte nuclear factor 4α(HNF4α) was detected by EMSA. 

Summary: Luteolin decreased the levels of HBsAg and HBeAg 

and the HBV DNA load both in the cultured medium and in 

serum, and it also suppressed the HBV DNA RI and the expression 

of HBsAg and HBcAg, which demonstrated that luteolin 

can effectively inhibit HBV replication both in vitro and in vivo. 

In addition, luteolin effectively downregulated the expression of 

HNF4α and decreased its binding level with HBV preC/C promoter 

in HepG2.2.15 cells. The extracellular signal-regulated 

kinase(ERK) and c-Jun N-terminal kinase(JNK) signal pathways, 

but not the p38 signal pathway in the MAPK family, were 

activated by luteolin, which can be blocked by their inhibitors, 

respectively. However, it was the ERK inhibitor(U0126) rather 

than the JNK inhibitor(SP600125) can attenuate the down-regulation 

of HNF4α by luteolin and restore the protein level of 

HNF4α. In addition, blocking the ERK pathway with inhibitor 

can cripple the inhibition of HBV replication by luteolin, while 

blocking the JNK pathway do not affect the inhibitory effect 

of luteolin on HBV replication. Conclusions: Luteolin inhibits 

HBV replication through down-regulating HNF4α expressin via 

activating ERK signal pathway. Keywords: luteolin; hepatitis B 

virus; replication; hepatocyte nuclear factor 4α; extracellular 

signal-regulated kinase 

Disclosures: 

The following authors have nothing to disclose: Yunhong Nong, Ying Shi, Miao 

Liu, Libo Yan, Yong Lin, Lang Bai, Hong Tang 

1563 

Redefining the Natural History of Chronic Hepatitis B: 

Using Viral Diversity to Classify Disease and Predict 

Serological Response to Nucleos(t)ide Analogue (NA) 

Therapy 

Julianne Bayliss 1 , Gillian Rosenberg 1 , Lilly Yuen 1 , Anuj Gaggar 2 , 

Kathryn M. Kitrinos 2 , Mani Subramanian 2 , Edward J. Gane 3 , 

Henry Lik-Yuen Chan 4 , Rachel Hammond 1 , Scott Bowden 1 , Peter 

A. Revill 1 , Stephen Locarnini 1 , Alex J. Thompson 5 ; 1 Molecular 

R&D, VIDRL, Melbourne, VIC, Australia; 2 Gilead Sciences, Foster 

City, CA; 3 New Zealand Liver Transplant Unit, Auckland City 

Hospital, Auckland, New Zealand; 4 Medicine and Therapeutics, 

Chinese University of Hong Kong, Hong Kong, Hong Kong; 5 Gastroenterology, 

St Vincent’s Hospital, Melbourne, VIC, Australia 

Background The presence of basal core promoter (BCP) and 

precore (PC) mutations in immune tolerant (IT) patients with 

chronic hepatitis B (CHB) suggests a transition between IT 

and immune clearance (IC) disease. Here, next generation 

sequencing (NGS) was used to analyse the impact of baseline 

viral diversity on CHB natural history and its association 

with baseline serology and response to NA treatment. Methods 

GS-US-203-0101 and GS-US-174-0103 evaluated tenofovir 

DF (TDF) containing regimens in IT and IC patients, respectively. 

Only genotype B/C patients were included (B= 83; 

C= 102). Patients were classified as True IT (ALT 20-40-80IU/mL). Viral diversity, defined as non-synonymous 

(NS), genotype specific viral substitutions from wild 

type (WT) and entropy scores across the whole genome (WG), 

were calculated using Illumina MiSeq NGS and correlated 

with baseline clinical and virological data and W192 serological 

response. Results 185 patients were analysed (baseline: 

median age 32yrs, 55% male; median ALT 29 IU/mL, HBV 

DNA 8.38 log 10 

IU/mL, HBsAg 51,590 IU/mL, HBeAg 3,358 

PEIU/mL). Increased viral diversity was associated with lower 

HBV DNA (p= 0.001), HBeAg (p< 0.0001) and HBsAg (p< 

0.0001) titres and higher ALT levels (p< 0.0001). Although 

HBV DNA, HBeAg and HBsAg titres did not differ significantly 

between True IT and Late IT patients (Table 1), a significantly 

higher percentage of Late IT patients had BCP and PC mutations 

(p= 0.001 and p< 0.0001, respectively). Late IT patients 

also had more NS substitutions (p= 0.001), higher entropy 

scores (p= 0.002), and were more likely to achieve HBeAg 

seroconversion by W192 compared to True IT patients (True 

IT= 3%, Late IT= 14%, p< 0.0001). Discussion There is a subset 

of IT CHB patients with a virological profile similar to IC 

patients. This subset of patients may be transitioning between 

IT and IC disease and are more likely to respond favourably 

to TDF therapy. 

Disclosures: 

Julianne Bayliss - Grant/Research Support: Gilead Inc. 

Gillian Rosenberg - Grant/Research Support: Gilead Sciences 



Sciences 

Mani Subramanian - Employment: Human Genome Sciences, Human Genome 

Sciences, Human Genome Sciences, Human Genome Sciences; Stock Shareholder: 

Human Genome Sciences, Human Genome Sciences, Human Genome 

Sciences, Human Genome Sciences 







Peter A. Revill - Grant/Research Support: Gilead Sciences 


Health 




Abbvie, BMS 

The following authors have nothing to disclose: Lilly Yuen, Rachel Hammond, 

Scott Bowden 

1564 

Angiopoetin-like protein 2 independently associated 

with liver inflammation and fibrosis in patients chronically 

infected with hepatitis B virus 

Hong Zhao, Yong-Qiong Deng, Gui-Qiang Wang; Department of 

infectious disease, Peking University First Hospital, Beijing, China 

Background: Guidelines recommend antiviral therapy for 

HBeAg(+)patients with HBV DNA >=20000IU/ml or HBeAg(-) 

patients with HBVDNA >=2000IU/ml, and alanine aminotransferase 

(ALT) levels more than 2 times of upper limit of normal 

(ULN). For patients who don’t meet the criteria above, therapy 

decision depends on hepatic histology, an invasive process. 

The available non-invasive assessments could only predict the 

stages of liver fibrosis. Therefore, a non-invasive method or 

biomarker predicting moderate and more inflammation or significant 

fibrosis is urgently needed. Angiopoetin-like protein 2 

(Angptl2) is a mediator of chronic inflammation contributing to 

extracellular matrix remodeling. Aim: The aim of the study was 

to explore the predicting value of serum angptl2 for moderate


and more inflammation or significant fibrosis. Methods: One 

hundred and seventy-three patients with chronic hepatitis B 

virus (HBV) infection, whose treatment decisions depended on 

liver biopsies, were enrolled in the study (NCT01962155 and 

ChiCTR-DDT-13003724). The levels of serum angptl2 were 

detected by Human ANGPTL2 Assay kit (Immuno-Biological 

Laboratories Co., Japan). Liver biopsies were performed in 

all patients. Histological Activity Index (HAI) and Liver fibrosis 

stage were assessed according to Ishak criteria independently 

by 2 pathologists. Results: 1) Serum Angptl2 levels were significantly 

associated with not only HAI scores (p=0.000, spearman 

rho=0.327) but also the liver fibrosis stages (P=0.000, 

spearman rho=0.318). 2) Patients with moderate and more 

inflammation or significant fibrosis (HAI>=5 or F>=3)who 

should be treated immediatelyhad higher serum angptl2 (5.76 

± 2.62 vs 4.31±1.82 ug/ml), AST, GGT and lower platelet 

counts, HBsAg. Multivariate analysis confirmed that only 

serum angptl2 level could independently predict urgent antiviral 

therapy. 3) Angptl2 showed areas under the receiver 

operating characteristics curve of 0.69 (95% CI: 0.60, 0.76) 

for indicating moderate and more inflammation or significant 

fibrosis. Leave-one-out cross-validation showed 64% cross-validation 

grouped cases correctly classified. Using a cutoff value 

of ≥7.36ug/ml, moderate and more inflammation or significant 

fibrosis could be correctly identified with 91.76% specificity 

and 72.0% PPV. Similarly, a lower angptl2 level (3.95 ug/ 

ml) could excluded urgent therapy with 78.41% specificity 

and 70.8% PPV. Conclusion: Serum angptl2, a novel single 

biomarker, independently predict moderate and more inflammation 

or significant fibrosis, saving half of patients from liver 

biopsies. 

Disclosures: 

The following authors have nothing to disclose: Hong Zhao, Yong-Qiong Deng, 

Gui-Qiang Wang 

1565 

PreS mutations analyzed by deep sequencing are 

correlated with the progression of liver diseases and 

HBsAg production. 

Yuichiro Suzuki, Shinya Maekawa, Nobutoshi Komatsu, Mitsuaki 

Sato, Masaru Muraoka, Shuya Matsuda, Mika Miura, Yasuhiro 

Nakayama, Taisuke Inoue, Minoru Sakamoto, Nobuyuki Enomoto; 

University of Yamanashi, Chuo, Japan 

Background and Aim: Recently, it is considered that achieving 

HBsAg seroclearance is the ultimate therapeutic goal in 

the treatment of chronic hepatitis B since serum HBsAg titer 

is suspected to reflect HBV-cccDNA titer in the liver. On the 

other hand, preS region of HBV genome responsible for HBsAg 

production was reported to be often mutated in advanced liver 

disease by direct sequencing. However, the interrelationship 

among HBsAg, preS mutation and liver disease progression is 

complicated and remains unclear. In the present study, to clarify 

the interrelationship among HBsAg titer, preS mutation and 

disease progression, we performed deep sequencing study 

for preS mutations. Methods: A total of 96 patients including 

32 inactive carriers (IC), 28 with chronic hepatitis (CH) and 

36 with cirrhosis or hepatocellular carcinoma (LC/HCC) were 

analyzed. All of inactive carriers were HBeAg negative, and 

22 patients were with HCC. They were all nucleotide analogue 

naïve. Deep sequencing was performed targeting 522nt in the 

preS region from patient serum, and proportion of preS1 and 

preS2 start codon mutations, preS1 and preS2 deletions was 

determined in each patient. Results: In IC group, CH group, 

and LC/HCC group, median HBsAg was respectively 688 IU/ 

ml, 2075 IU/ml, and 1065IU/ml. HBsAg level


ent association of the genotype 5 isolates with region/country 

of birth. Conclusion: This study demonstrates the increasing 

proportion of HDV infections which were acquired overseas 

prior to migration and compares phylogenetic analysis with 

country of birth data from surveillance notifications. Detection 

of genotype 5 strains (predominantly found in African patients) 

are increasing in Australia, reflecting changing migration patterns. 

Understanding the genotypes and country of origin of 

HDV may play a role in the diagnosis and management of an 

infection that is poorly recognized. 

Disclosures: 


Health 

The following authors have nothing to disclose: Kathy Jackson, Margaret Littlejohn, 

Lilly Yuen, Benjamin C. Cowie, Jennifer H. MacLachlan, Scott Bowden 

1567 

Kinetics of Serum Hepatitis B Surface Antigen During 

the First Year of Tenofovir Treatment in Patients With 

Chronic Hepatitis B 

Choong Kyun Noh, Soon Sun Kim, Sun Young Park, Joohan Park, 

Hyo Jung Cho, Jae Youn Cheong, Sung Won Cho; Ajou University 

School of Medicine, Suwon, Korea (the Republic of) 

Aims The predictive role of quantitative hepatitis B surface antigen 

(HBsAg) levels for HBsAg or hepatitis B e antigen (HBeAg) 

loss/seroconversion in patients receiving potent oral antiviral 

therapy is suggested. However, data on patients with genotype 

C hepatitis B virus is limited. We evaluated the HBsAg 

kinetics in chronic hepatitis B patients treated with tenofovir 

and correlation with treatment response. We also compared 

HBsAg kinetics with those of chronic hepatitis B patients 

treated with entecavir. Methods Serial HBsAg levels were 

determined in sera from 66 tenofovir (29 HBeAg-positive and 

37 HBeAg-negative) and 101 entecavir (59 HBeAg-positive 

and 42 HBeAg-negative)-treated chronic hepatitis B patients at 

baseline, 3, 6 and 12 months during treatment. Results During 

12 months of tenofovir treatment, the mean HBsAg levels did 

not significantly decrease in both HBeAg-positive and negative 

patients (3.40, 3.24, 3.26 and 3.30 log 10 

IU/ml and 3.31, 

3.39, 3.37 and 3.34 log 10 

IU/ml at baseline, 3, 6, 12 months 

of treatment, respectively). Only two HBeAg-positive patients 

exhibited a significant (≥ 1.0 log) decline in HBsAg levels 

until 12 months. However, decline of HBsAg levels between 

baseline and 3 months of treatment showed a tendency to be 

larger in HBeAg-positive patients than those in HBeAg-negative 

patients (0.09 log 10 

IU/ml decrement vs. 0.07 log 10 

IU/ 

ml increment, P = 0.66). The on-treatment HBsAg levels did 

not correlate with HBeAg loss/seroconversion at 12 months in 

HBeAg-positive tenofovir-treated patients. Changes of HBsAg 

levels during tenofovir and entecavir were not significantly different 

in both HBeAg-positive and HBeAg-negative patients. 

Conclusions Decline of HBsAg levels during the 12 months 

of tenofovir treatment was insignificant in chronic hepatitis 

B patients with genotype C. Long-term oral antiviral therapy 

will be needed in majority of chronic hepatitis B patients even 

though antiviral therapies are very potent. 

Disclosures: 

The following authors have nothing to disclose: Choong Kyun Noh, Soon Sun 

Kim, Sun Young Park, Joohan Park, Hyo Jung Cho, Jae Youn Cheong, Sung 

Won Cho 

1568 

The PAGE-B Score Stratifies Chronic Hepatitis B Patients 

with Compensated Cirrhosis at High Risk of Hepatocellular 

Carcinoma Development with Good Accuracy 

Willem Pieter Brouwer 1 , Bettina E. Hansen 1 , Elena Raffetti 2 , 

Francesco Donato 2 , Giovanna Fattovich 3 ; 1 Gastroenterology & 

Hepatology, Erasmus Medical Center Rotterdam, Rotterdam, Netherlands; 

2 Epidemiology and Public Health, Institute of Hygiene, 

Brescia, Italy; 3 Medicine, Azienda Ospedaliera Universitaria Integrata, 

Verona, Italy 

Background & aims. The PAGE-B score has been associated 

with HCC development in Caucasian patients treated with 

potent nucleos(t)ide analogues. We aimed to assess the performance 

of this marker in a cohort of untreated Western chronic 

hepatitis B (CHB) patients with Child Pugh A cirrhosis. Methods. 

In this retrospective cohort study conducted at 9 tertiary care 

centres in Europe, CHB patients with biopsy-proven, compensated 

cirrhosis and available laboratory measurements were 

included. The PAGE-B, CU-HCC, GAG-HCC and REACH-B 

scores were calculated and associated with the occurrence 

of liver failure, HCC development, transplant-free survival or 

any clinical event (combined endpoint including all the respective 

events). Results. Of 134 patients, mean age at inclusion 

was 47±13 years, 70% was HBeAg-negative and 87% male. 

In total, 16 developed an HCC, 21 died or underwent liver 

transplantation and 41 developed any clinical event during 

a median follow-up of 6.2 years (IQR 3.1-8.6). The 5-year 

(95%CI) HCC-free and transplant-free survival was 90% (84- 

95) and 87% (81-93), respectively. The PAGE-B score was 

significantly associated with HCC development (HR 1.34, 

95%CI:1.2-1.6, p


1569 

Health care disparity in delivering optimal care to 

chronic hepatitis B pregnant mothers 

Rasham Mittal, Amandeep K. Sahota; Kaiser Permanente Los 

Angeles Medical Center, Los Angeles, CA 

Background Roughly 24,000 chronic hepatitis B (CHB) pregnant 

women give birth in United States every year. Since perinatal 

acquisition of CHB contributes significantly to prevalence 

of CHB, it becomes imperative to provide optimal antenatal 

care to these mothers. Study Aim: - Determine if health care 

disparities exist in antenatal care of CHB pregnant mothers at 

a large integrated health care system in United States. Methods: 

Retrospective study (January 2005- June 2010) at a large 

integrated health care system in Southern California. All CHB 

pregnant mothers identified using the regional perinatal database. 

Inclusion criteria: age > 18 years and positive serum 

HBsAg (hepatitis B surface antigen). They were excluded if 

tested negative for HBsAg; tested positive for preliminary 

enzyme-linked immunosorbent assay (ELISA) HBsAg but had 

non-reactive HBsAg neutralization confirmatory antibody assay 

(false positive); termination of pregnancy or lost to follow up. 

Only first live-birth pregnancy was considered for CHB mothers 

who delivered more than once during the study timeline. Chart 

review was performed to determine maternal antenatal hepatitis 

Be antigen (HBeAg) serology, antenatal hepatitis B virus (HBV) 

viral load and specialist consultation visits. Results: A total of 

462 CHB pregnant mothers were identified. The mean maternal 

age in years was 32.5 ± 4.8 (range 18-45, median 33). 

Highest prevalence of CHB was noted in Asian Pacific Islander 

mothers (78.8%), followed by non-Hispanic white (9.7%), Hispanic 

white (5.8%), non-Hispanic black (5.2%) and Hispanic 

black (0.4%). Antenatal hepatitis Be antigen (HBeAg) serology 

testing was performed in 274 (59.3%) and hepatitis B virus 

(HBV) viral load was tested in 156 (33.8%) mother only. Only 

93 (20.1%) mothers had a specialist consultation during their 

pregnancy. Among these 93 mothers, 90 consulted gastroenterologist 

and 3 hepatologist. Additionally 104 (26.2%) infants 

born to these CHB mothers had no HBsAg serological testing 

at 9-18 months follow up. Conclusion: - Significant health care 

disparities in standard care of CHB pregnant mothers identified 

at a large integrated health care system in United States. 

Disclosures: 

The following authors have nothing to disclose: Rasham Mittal, Amandeep K. 

Sahota 

1570 

Ultrasound predicts steatosis in patients with chronic 

hepatitis B 

Erin Kelly 2,1 , Vickie A. Feldstein 3 , Rebecca Hudock 1 , Marion G. 

Peters 1 ; 1 Hepatology, University of California San Francisco, San 

Francisco, CA; 2 Gastroenterology and Hepatology, University of 

Ottawa, Ottawa, ON, Canada; 3 Radiology, UCSF, San Francisco, 

CA 

Inflammation and fibrosis may impair the ability of ultrasound 

to identify steatosis in patients with chronic hepatitis B (CHB). 

We determined the accuracy of ultrasound (US) in grading 

steatosis in patients with CHB compared to liver biopsy, and 

examined clinical factors associated with steatosis. This was a 

single-center, retrospective study of all non-transplanted CHB 

patients undergoing US and same day liver biopsies from 

2004-2014. Steatosis was graded by US as none, mild, moderate 

or severe. Liver histology graded steatosis (0:


greatest risk. Here we investigate whether a qHBsAg threshold 

(


1573 

Application of Highly Sensitive Chemiluminescent 

Enzyme Immunoassay for Hepatitis B Surface Antigen to 

Detect Occult HBV infection: an Appropriate Method as 

Pre-Transfusion Testing. 

Takako Inoue 1 , Noriyo Ochi 1 , Takaaki Goto 1 , Shintaro Ogawa 2 , 

Noboru Shinkai 3 , Kumiko Ohne 1 , Yukio Wakimoto 1 , Yasuhito 

Tanaka 2,1 ; 1 Clinical Laboratory, Nagoya City University Hospital, 

Nagoya, Japan; 2 Department of Virology & Liver unit, Nagoya City 

University Graduate School of Medical Sciences, Nagoya, Japan; 

3 Department of Gastroenterology and Metabolism, Nagoya City 


Background: We developed highly sensitive chemiluminescent 

enzyme immunoassay (CLEIA) for detection of hepatitis B 

surface antigen (HBsAg). As recently reported, this assay uses 

for quantitative HBsAg detection by combining monoclonal 

antibodies, each specific for a different epitope of the antigen, 

and employing an improved conjugation technique. Here, 

we demonstrate that highly sensitive HBsAg CLEIA (Lumipulse 

HBsAg-HQ) is an appropriate and precise way as pre-transfusion 

testing to detect occult HBV viremia. Methods: In Japan, 

two quantitative HBsAg detection systems are available: Architect 

HBsAg-QT (Abbott Japan) (detection range, 50 to 250,000 

mIU/mL) and HISCL HBsAg (Sysmex) (detection range, 30 to 

2,500,000 mIU/mL). The sensitivity of highly sensitive HBsAg 

CLEIA (Lumipulse HBsAg-HQ) (Fujirebio, Inc.) (5150,000,000 

mIU/mL) is approximately 6-fold higher than that of HISCL 

HBsAg (30 mIU/mL). In this study, the performance of Lumipulse 

HBsAg-HQ was compared with that of HISCL HBsAg. 

This study protocol was approved by the appropriate institutional 

ethics review committees. Results: As pre-transfusion 

testing, 4,733 serum samples were measured HBsAg by HISCL 

HBsAg in our hospital between July 1st 2013 and February 

28th 2015. Of the 4,733 samples, 117 samples (2.47%) were 

HBsAg seronegative but showed low concentration of HBsAg 

(10 to 20 mIU/mL) as determined by HISCL HBsAg. Of the 

117 samples, 17 (14.5%) were positive for anti-hepatitis B 

core antigen (anti-HBc), and 15 of the 17 samples were available 

for additional assay. They were measured by Lumipulse 

HBsAg-HQ retrospectively and 3 (20%) were detectable for 

HBsAg. Two samples were from chronic hepatitis B carriers 

with HBsAg seroclearance and one was from an adult T-cell 

leukemia/lymphoma patient who was diagnosed as HBV reactivation. 

Their HBsAg concentrations by Lumipulse HBsAg-HQ 

were 8.0, 38.5 and 7.2 mIU/mL, respectively. Conclusion: As 

pre-transfusion testing, Lumipulse HBsAg-HQ is very appropriate 

and precise system for detection of occult HBV viremia as 

well as HBV reactivation at early phase. 

Disclosures: 



Squibb 

The following authors have nothing to disclose: Takako Inoue, Noriyo Ochi, 

Takaaki Goto, Shintaro Ogawa, Noboru Shinkai, Kumiko Ohne, Yukio 

Wakimoto 

1574 

Collaborative Public Health Efforts to Test and Link Foreign-Born 

Persons with Chronic Hepatitis B Virus Infection 

to Care 

Aaron M. Harris, Ben Schoenbachler, Gilberto Ramirez, Claudia 

Vellozzi, Geoff Beckett; Division of Viral Hepatitis, Centers for 

Disease Control and Prevention, Atlanta, GA 

Background: Hepatitis B virus (HBV) infection continues to 

be a public health threat. An estimated 1 million persons are 

chronically infected with HBV in the United States. Of these, 

70% are foreign-born and more than two-thirds are unaware 

of their infection. We launched a public health initiative among 

foreign-born persons for HBV testing and linkage to care. 

Methods: Nine U.S. sites participated in the initiative during 

October 2012–September 2014. Sites partnered with healthcare 

centers and community-based organizations for recruitment 

of foreign-born persons. Blood samples were collected 

and tested for hepatitis B surface antigen (HBsAg). Information 

regarding risk factors, follow-up counseling, specialty referrals, 

and medical appointment attendance was collected. Results: 

Of 23,144 individuals tested across all sites, 1,317 (5.7%) 

were HBsAg-positive. Of these, median age was 47 years, 

91% had at least one risk factor for HBV infection, 85% were 

provided counseling, 83% were referred to care, and 46% 

attended a first medical visit. The proportion of HBsAg-positive 

persons by region of origin included: Africa (9.7%), Western 

Pacific (6.4%), Eastern Mediterranean (5.2%), South-East Asia 

(4.9%), South America (2.4%), Eastern Europe (2.3%), and 

North America (0.9%). Conclusions: Community-based HBV 

testing initiatives can identify substantial numbers of persons 

with chronic HBV infection. However, strategies are needed to 

improve linkage to HBV-directed medical care for foreign-born 

individuals living with chronic HBV infection. 

Disclosures: 

The following authors have nothing to disclose: Aaron M. Harris, Ben Schoenbachler, 

Gilberto Ramirez, Claudia Vellozzi, Geoff Beckett 

1575 

Hepatitis B viral load in dried blood spots: a validation 

study in Zambia 

Michael J. Vinikoor 1,2 , Samuel Zürcher 3 , Kalo Musukuma 4,2 , Obert 

Kachuwaire 2 , Andri Rauch 3 , Benjamin Chi 5 , Meri Gorgievski 3 , 

Marcel Zwahlen 3 , Gilles Wandeler 3 ; 1 Medicine, University of 

North Carolina at Chapel Hill, Lusaka, Zambia; 2 Centre for Infectious 

Disease Research in Zambia, Lusaka, Zambia; 3 University of 

Bern, Bern, Switzerland; 4 University of Zambia, Lusaka, Zambia; 

5 Obstetrics & Gynecology, University of North Carolina at Chapel 

Hill, Chapel Hill, NC 

Study Purpose: Access to hepatitis B viral load (VL) testing is 

poor in sub-Saharan Africa (SSA) due to economic and logistical 

reasons. In a laboratory in Lusaka, Zambia, we compared 

hepatitis B VLs between plasma samples and dried blood spots 

(DBS), a potential alternative specimen type for such settings. 

Methods: Paired plasma and DBS samples from HIV-hepatitis B 

virus (HBV) co-infected Zambian adults were analyzed for HBV 

VL using COBAS AmpliPrep/COBS Taqman HBV test version 

2.0 and genotype by direct sequencing. We used Bland-Altman 

analysis to compare VLs between DBS and plasma and by 

HBV genotype. Logistic regression analysis was conducted to 

assess the probability of an undetectable DBS result by plasma 

VL. Results: Among 68 participants, median age was 34 years, 

61.8% were men, and plasma HBV VLs ranged from 170,000,000 IU/ml. Among sequenced viruses, 28 were 

genotype A1 and 27 were genotype E. Bland-Altman plots 

suggested strong agreement between DBS and plasma VLs (see 

Figure). On average VLs were 1.59 log 10 

IU/ml (95% CI: 1.48 

to 1.70) lower in DBS compared to plasma. The probability of 

an undetectable DBS result at a plasma VL of ≥2,000 IU/ml 

was 1.8% (95% CI: 0.5-6.6) and 0.2% (95% CI: 0.03-1.7) at 

≥20,000 IU/ml. Conclusions: We demonstrated the feasibility 

and performance of HBV VL testing with DBS in a Zambian 

laboratory and observed a strong agreement between sample 

types and high sensitivity in DBS at plasma VL ≥2,000 IU/ml.


As HBV treatment expands, DBS could increase access to HBV 

VL testing in SSA settings. 

Bland-Altman plot of the agreement between HBV viral loads in 

plasma and dried blood spots 

Disclosures: 

Andri Rauch - Advisory Committees or Review Panels: Gilead Sciences, Abbvie, 

MSD, Janssen Cilag 

The following authors have nothing to disclose: Michael J. Vinikoor, Samuel 

Zürcher, Kalo Musukuma, Obert Kachuwaire, Benjamin Chi, Meri Gorgievski, 

Marcel Zwahlen, Gilles Wandeler 

1576 

Utility of quantitative hepatitis B surface antigen (qHBsAg) 

testing for predicting maternal viremia associated 

with mother to child transmission of HBV in a multiethnic 

cohort of pregnant chronic hepatitis B (CHB) carriers 

in Canada 

Golasa Samadi Kochaksaraei 1 , Carmen L. Charlton 2 , Stephen 

Congly 1 , Trudy Matwiy 1 , Eliana Castillo 1 , Steven R Martin 3 , Carla 

S. Coffin 1 ; 1 Medicine, University of Calgary, Calgary, AB, Canada; 

2 Provincial Laboratory for Public Health (ProvLab), University 

of Alberta Hospital, Edmonton, AB, Canada; 3 Pediatrics, Alberta 

Children’s Hospital, University of Calgary, Calgary, AB, Canada 

Background and Aims: Mother-to-child transmission of hepatitis 

B despite complete passive-active immunoprophylaxis is linked 

to high maternal viremia. Available data from our clinic and 

others suggest that anti-HBV therapy (i..e, Tenofovir, TDF) in 

highly viremic (HBV DNA ≥7 log IU/ml) mothers is safe and 

can reduce HBV DNA levels to below the threshold associated 

with vaccine failure. Quantitative HBsAg (qHBsAg) monitoring 

is increasingly used in management of chronic hepatitis B 

(CHB), but there is limited data in pregnancy. We conducted 

a prospective observational study to determine whether qHBsAg 

can be used as a valid surrogate marker of HBV DNA. 

Methods: CHB pregnant patients were recruited from a hepatology 

outpatient practice or an obstetrics internal medicine 

clinic. Demographics and laboratory data (i.e., HBeAg and 

ALT), HBV DNA and quantitative HBsAg were assessed in the 

second-third trimester. Statistical analysis was performed by 

Spearman’s rank correlation and student’s t-test. Results: In 96 

pregnant CHB patients enrolled to date (of which 4 received 

TDF therapy), median age 32 years (IQR 28-35), 66% Asian, 

23% African, and 11% other (Hispanic or Caucasian). Overall, 

22% (21/96) were HBeAg (+), median ALT was 18.5 

U/L (IQR 14-31), median HBV DNA was 2.46 log IU/ml (IQR 

1.44-3.54). In 69/96 with qHBsAg testing to date, there was a 

statistically significant difference in qHBsAg in HBeAg positive 

vs. HBeAg negative CHB patients (3.58 log IU/ml [IQR 2.76- 

4.28] vs 3.38 log IU/ml [IQR 2.84-3.77], p=0.007) and HBV 

DNA (2.57 log IU/ml [IQR 0-7.89] in HBeAg positive vs 2.1 

log IU/ml [IQR 1.56-2.92] in HBeAg negative, p=0.0001). 

In HBeAg positive patients, there was a significant correlation 

between qHBsAg titer and HBV DNA level (r=0.508, p=0.013) 

whereas no significant correlation was noted in HBeAg negative 

group (r=0.105, p=0.39). In receiver operating characteristic 

(ROC) analysis, the optimal qHBsAg cut-off values 

for predicting HBV DNA level believed to be associated with 

immunoprophylaxis failure (i.e., HBV DNA ≥7 log IU/ml) was 

3.58 log IU/ml (accuracy 85.5%, sensitivity 87.5%, specificity 

80%). Conclusion: Quantitative HBsAg positively correlates 

with serum HBV DNA level in HBeAg positive CHB pregnant 

patients and could also be used as a marker for high maternal 

viremia, and the need for anti-HBV NA therapy to prevent HBV 

immunoprophylaxis failure. 

Disclosures: 

Carla S. Coffin - Advisory Committees or Review Panels: Janssen, GSK; Grant/ 

Research Support: BMS, Gilead Sciences, Roche 

The following authors have nothing to disclose: Golasa Samadi Kochaksaraei, 

Carmen L. Charlton, Stephen Congly, Trudy Matwiy, Eliana Castillo, Steven R 

Martin 

1577 

Nationwide Prospective Survey in Japan to Clarify the 

Incidence of Viral Reactivation in Patients with Prior 

HBV Infection Receiving Immunosuppressive Agents 

Masamitsu Nakao 1 , Nobuaki Nakayama 1 , Yoshihito Uchida 1 , 

Masashi Mizokami 2 , Satoshi Mochida 1 ; 1 Gastroenterology 

& Hepatology, Saitama Medical University, Saitama, Japan; 

2 Research Center for Hepatitis and Immunology, National Center 

for Global Health and Medicine, Ichikawa, Japan 

Aim: More than 20% of the population of individuals over 50 

years of age in Japan have a history of transient HBV infection. 

In such patients, serum HBV-DNA may become detectable following 

immunosuppressive therapies, potentially associated 

with the development of severe hepatitis. While HBV reactivation 

is known to occur at a high incidence in patients receiving 

rituximab, the incidence in those with immunosuppressive 

agents are yet to be investigated. We conducted 2 types 

nationwide survey to determine the incidence of HBV reactivation. 

Methods: A total of 420 patients from 59 institutions, who 

tested negative for serum HBs-antigen, but positive for serum 

anti-HBc and/or anti-HBs antibodies were enrolled, and were 

givin immunosuppressive agents such as glucocorticoids (≥0.5 

mg/kg of prednisolone), methotrexate and/or biologic agents. 

Among them, 131 patiens received the therapies following 

the enrollement (Part-1), and the remaining 289 patients were 

enrolled at 6 months or later after the initiation of the therapy 

when HBV reactivation has not yet developed (Part-2). In all 

patients serum HBV-DNA levels were measured evry 1 month, 

and the incidences if HBV reactivation, defined as the increase 

of serum HBV-DNA levels of 2.1 Log copies/mL or more, were 

evaluated by Kaplan-Meier method; within 6 months after the 

initiation of therapies in patients in Part-1 study, and later than 

6 months in those enrolled in both Part-1 and Part-2 studies. 

Results: HBV reactivation was found in 5 patients in Part-1 

study during mediam periods for 38 (1-62) months, and in 5 

patients in Part-2 study after the initiarion of therapies from 7 

to 341 months. HBV reactivation occurred even in those who 

received monotherapy with methotrexate or glucocorticoid as 

well as the therapies with biologic agents. These patients were 

given entecavir at least within 1 month after HBV reactivation, 

and liver injuries did not occur in all of them. Kaplan-Meier


analysis revealed that the cumulative incidences of HBV reactivation 

were 0.8%, 3.2% and 3.2% at 1, 3 and 6 months, 

respectively, after the intiation of therapies through Part-1 

study. Also, the analysis thorugh both Part-1 and Part-2 studies 

demonstrated that the increase of incidence later than 6 

months were 0%, 0.5%, 1.5% and 1.5% at 12, 24, 36 and 48 

months, respectively. Conclusion: HBV reactivation developed 

mainly within 6 months after initiation of immunosuppressive 

therapies, and the risk was reduced then later. Monitoring of 

HBV reactivation by serum HBV-DNA measurement is recommended 

to be done especially within 6 months after initiation 

of immonusuppresive therapies and/or alteration of agents for 

the therapies. 

Disclosures: 




The following authors have nothing to disclose: Masamitsu Nakao, Nobuaki 

Nakayama, Masashi Mizokami 

1578 

HBV-related cirrhosis in the Chronic Hepatitis Cohort 

Study (CHeCS) 

Stuart C. Gordon 2,3 , Loralee B. Rupp 1 , Joseph A. Boscarino 4 , 

Mark A. Schmidt 5 , Connie M. Trinacty 6 , Lois Lamerato 7 , Nancy 

Oja-Tebbe 7 , Mei Lu 7 ; 1 Center for Health Policy & Health Services 

Research, Henry Ford Health System, Detroit, MI; 2 Division of Gastroenterology 

and Hepatology, Henry Ford Health System, Detroit, 

MI; 3 School of Medicine, Wayne State University, Detroit, MI; 

4 Center for Health Research, Geisinger Health System, Danville, 

PA; 5 Center for Health Research, Kaiser Permanente Northwest, 

Portland, OR; 6 Center for Health Research, Kaiser Permanente 

Hawaii, Honolulu, HI; 7 Public Health Sciences, Henry Ford Health 

System, Detroit, MI 

PURPOSE: The overall spectrum of liver disease among Americans 

with chronic hepatitis B (CHB) infection remains unknown, 

and staging liver biopsy has become increasingly uncommon. 

We used four different methods, including liver biopsy, to 

identify potential cirrhosis among CHB patients enrolled in the 

Chronic Hepatitis Cohort Study (CHeCS), an ongoing observational 

study at four large, integrated health systems in the 

United States. METHODS: We included patients who met predefined 

inclusion criteria for CHB whose case status had been 

confirmed through chart abstraction. We excluded patients 

coinfected with hepatitis C and liver transplant recipients. Data 

were collected through 2012 from liver biopsy reports, lab 

results, and diagnosis/procedure codes contained in the electronic 

health record. We calculated FIB-4 scores based on most 

recently available aminotransferase and platelet count values 

collected when a patient was not on antiviral therapy. Cirrhosis 

was identified via four methods: (1) liver biopsy reports, (2) 

presence of a diagnosis code for cirrhosis (ICD-9 diagnosis 

codes 571.2 and 571.5), (3) presence of a diagnosis or procedure 

code for end stage liver disease (ESLD), and (4) FIB-4 

score >5.17, a cut-off value previously validated as predictive 

of cirrhosis in this cohort. We compared the extent to which 

cirrhosis was indicated by each method individually and by 

multiple methods. RESULTS: Among the 2,731 CHB patients, 

24% were 60 years old, 54% were male, and 

53% were Asian. Median follow-up time was 6.0 years. Of the 

total, 564 (21%) patients had at least one biopsy report during 

follow-up, and of those, 85 (15% of those who had a biopsy) 

had a recent biopsy less than two years old. Overall, cirrhosis 

was indicated for 520 (19%) patients by at least one of the 

four methods: by liver biopsy for 66 patients (2% of patients 

overall, and 12% among those with any liver biopsy report); 

by FIB-4 score for 281 (10%); by diagnosis code for cirrhosis 

for 318 (12%); and by diagnosis or procedure code for ESLD 

for 216 (8%), 145 of whom had a diagnosis code for both cirrhosis 

and ESLD. Of the 281 patients with FIB-4 score >5.17, 

only 26 (9%) had been diagnosed with cirrhosis via biopsy, 

135 (48%) via ICD-9 code for cirrhosis, and 113 (40%) via 

diagnosis or procedure code indicating ESLD. CONCLUSION: 

An estimated 19% of CHB patients among our cohort had 

indication of cirrhosis at some point in their follow-up through 

2012. The use of additional parameters, including a previously 

validated biomarker as a surrogate for more advanced liver 

disease, potentially identifies additional unrecognized cirrhosis 

in untreated CHB patients. 

Disclosures: 




The following authors have nothing to disclose: Loralee B. Rupp, Joseph A. Boscarino, 

Mark A. Schmidt, Connie M. Trinacty, Lois Lamerato, Nancy Oja-Tebbe, 

Mei Lu 

1579 

HBsAg transferring from mother to infant through placenta 

account for non-response to hepatitis B vaccine in 

infant from HBsAg(+) mother 

Jing Wang 1 , Jinfeng Liu 2 , Yingli He 2 , Dongfang Jin 3 , Yuan Yang 2 , 

Li Jin 1 , Taotao Yan 1 , Furong Cao 1 , Shulin Zhang 2 , Yingren Zhao 2 , 

Tianyan Chen 2 ; 1 Xi’an Jiaotong University, Xi’an, China; 2 The First 

Affiliated Hospital of Medical College, Xi’an Jiaotong University, 

Xi’an, Shaanxi Province, China, Xi’an, China; 3 Shaanxi Kangfu 

Hospital, Xi’an, China 

Background: It is common for infants from HBsAg(+) mothers 

to fail to mounting protective immunity after standard hepatitis 

B vaccination and then develop infection. Maternal HBV 

markers crossing placenta may lead to neonatal temporal HBV 

markers positivity rather than infection, whether these markers 

affecting infantile HB vaccine response remains unclear. Methods: 

A total of 328 HBsAg(+) mothers and their offspring were 

enrolled. All infants received combined immunoprophylaxis 

and then followed up to 12 months. HBsAg, anti-HBs titer and 

HBV DNA load were assayed. Immunohistochemical staining 

for HBsAg in placenta was employed to explore HBsAg transferring. 

Results: Firstly, 22 (6.7%) were non-responders to HB 

vaccine were identified from those 328 infants. By stastical 

analysis, we found that: 1. HBsAg(+) newborns showed higher 

risk of non-response than HBsAg(−) infants ( 13.0% vs. 5.0%, 

P = 0.016). 2. Infants from high HBsAg titer mothers displayed 

higher risk of HBsAg positive at birth than those from low 

titer mothers (45.3% vs. 2.8%, P < 0.001). 3. HBsAg titer in 

mothers of HBsAg(+) newborns was much higher than mothers 

of HBsAg(−) newborn (P < 0.001). All those data supporting 

HBsAg can be transferring through placenta. Moreover, 

our hypothesis was reinforced by immunostaining with specific 

antibody against HBsAg, showing placenta of HBsAg(+) 

infants demonstrated higher prevalence (87.5% vs. 30.8%, 

P = 0.024) and stronger immunostaining (P = 0.008) than 

placenta of HBsAg(−) infants. Conclusion: HBsAg can transfer 

though placenta from mother to infant and the neonatal HBsAg 

positivity was associated with non-response to HB vaccine. 

Disclosures: 

The following authors have nothing to disclose: Jing Wang, Jinfeng Liu, Yingli 

He, Dongfang Jin, Yuan Yang, Li Jin, Taotao Yan, Furong Cao, Shulin Zhang, 

Yingren Zhao, Tianyan Chen


1580 

Aldehyde dehydrogenases-2 rs671 polymorphism is 

associated with an increased risk of hepatocellular carcinoma 

in patients with hepatitis B virus 

Jing Sun 1 , Hongqin Xu 1 , Xiuting He 1 , Jingyun Wang 1 , Xiaomei 

Wang 1 , Bin Gao 2 , Junqi Niu 1 , Yanhang Gao 2 ; 1 The first hospital 

of Jilin University, Changchun, China; 2 National Institutes of Health 

NIH/NIAAA, Rockville, MD 

Background and aims: Hepatocellular carcinoma (HCC) is 

the second-most common cancer in China and accounts for 

51% of the deaths from liver cancer worldwide. Up to 80% of 

HCC cases in China are attributable to hepatitis B virus (HBV), 

but the exact pathogenesis is still not completely understood. 

Aldehyde dehydrogenase 2(ALDH2) is well known about 

its role on detoxifying aldehydes in ethanol metabolism. An 

ALDH2 inactivating mutation is the most common single point 

mutation in humans, mostly found in East Asians. There is a 

good geographical correlation between the prevalence of the 

ALDH2 gene mutation and HBV infection, but the relationship 

between these two factors is unclear. Methods: In the present 

study, we investigated the association between ALDH2 (rs671, 

rs10849970 and rs886205) polymorphisms, chronic alcohol 

consumption, and the risk of HBV-related HCC among Han 

populations in Northeast China in a case-control study (2012- 

2015). A total of 930 subjects were included in the study, 

including 102 cases with HBV-related chronic hepatitis, 264 

cases with HBV-related cirrhosis, 281 cases with HBV-related 

HCC, and 283 healthy individuals as controls. The odds ratios 

(OR) and corresponding 95% confidence intervals (CI) were 

calculated using logistic regression models. Results: ALDH2 

rs671 includes three genotypes, GG (wild type), AA (homozygous 

mutation), GA (heterozygous mutation). The distribution 

difference of genotype frequencies of ALDH2 rs671 in the four 

groups we studied was statistically significant (P


between selected parameters of lipid metabolism and hepatic 

steatosis measured using CAP option of transient elastography 

(FibroScan), serum HBV-DNA and HBsAg levels during treatment 

with tenofovir (TDF), after switching from any antiHBV 

nucleos(t)ide analogue (NUC). Methods: The study included 

45 HBsAg-positive and HBeAg-negative pts, who within last 

3 months have been switched to TDF due to the lack of efficacy 

or intolerance to prior treatment. We evaluated the level 

of total cholesterol, low-density lipoprotein (LDL), high-density 

lipoprotein (HDL), triglycerides (TG), HBV-DNA and HBsAg 

levels. Liver steatosis and fibrosis were assessed FibroScan with 

CAP option. In statistical analysis parametric Pearson correlation 

and Rang Spearman correlation were used. Results were 

considered statistically significant if p


and 6.15-log copies/mL (4.0-above 8.1) among HBeAg-negative 

patients. Among HBeAg positive and negative patients, 

the percent ratios of cases whose HBV DNA concentration 

fell below 3-log copies/mL were 29.0/60.7 at 3 months, 

58.1/96.4 at 6 months, 87.1/100 at 12 months, 90.3/100 

at 18 months, and 90.3/100 at 24 months of therapy. The percent 

ratios of cases whose HBV DNA levels were undetectable 

were 0/7.1 at 3 months, 3.2/17.9 at 6 months, 12.9/23.1 

at 12 months, 9.7/53.6 at 18 months, and 25.8/71.4 at 24 

months of therapy. (2) A strong correlation of HBsAg values 

between the Architect and Lumipulse assays was observed in 

both HBeAg-positive and HBeAg-negative patients (r=0.88 and 

0.93, respectively). However, more than 1-log reduction of 

HBsAg values (IU/mL in the Architect and cutoff index [COI] in 

the Lumipulse) after 24 months of therapy was observed in only 

11.9% of samples by either assay. Conclusion: HBsAg values 

measured by the Architect and Lumipulse assays were directly 

correlated. However, HBsAg values decreased slowly during 

the 2-year ETV therapy whereas HBV DNA concentrations 

decreased more than 3-log copies/mL in all patients. Thus, 

neither of these assays are effective predictors of the efficacy 

of short-term ETV therapy. 

Disclosures: 

The following authors have nothing to disclose: Kazuma Shinkai, Hisashi Ishida, 

Ryousuke Kiyota, Taku Tashiro, Kentarou Nakagawa, Keisuke Fukutomi, Akio 

Ishihara, Tetsuya Iwasaki, Ryuichiro Iwasaki, Kumiko Nishio, Yuko Sakakibara, 

Takuya Yamada, Shoichi Nakazuru, Eiji Mita 

qHBsAg: -0.33 log IU/mL vs. -0.06 log IU/mL, p=0.02; Figure). 

The area under the curve for a prediction model based 

on week 12 for HBcrAg, ALT, and qHBsAg was 0.70 (CI-95% 

0.57-0.83, p=0.005). Conclusion. On-treatment HBcrAg level 

decline in the first 12 weeks is associated with response to 

peginterferon in HBeAg-negative CHB patients. HBcrAg levels 

are strongly correlated with HBV DNA. 

Disclosures: 





Roche, Gilead 






The following authors have nothing to disclose: Margo J. van Campenhout, Willem 

Pieter Brouwer, Vincent Rijckborst, Bettina E. Hansen 

1585 

Hepatitis B Core-Related Antigen Level Decline in the 

First 12 Weeks of Peginterferon Treatment is Associated 

with Response in HBeAg-Negative Chronic Hepatitis B 

Margo J. van Campenhout 1 , Willem Pieter Brouwer 1 , Vincent Rijckborst 

1 , Robert J. de Knegt 1 , Andre Boonstra 1 , Harry L. Janssen 2,1 , 

Bettina E. Hansen 1,3 ; 1 Gastroenterology & Hepatology, Erasmus 

MC University Medical Center Rotterdam, Rotterdam, Netherlands; 

2 Toronto Centre for Liver Disease, Toronto Western and General 

Hospital, University Health Network, Toronto, ON, Canada; 3 Public 

Health, Erasmus MC University Medical Center Rotterdam, Rotterdam, 

Netherlands 

Background. Markers for prediction of response in HBeAg-negative 

chronic hepatitis B (CHB) patients are scarce. Hepatitis B 

core-related antigen (HBcrAg), which is a new serum marker 

for the combined measure of HBcAg, HBeAg, and p22cr, 

may be useful for response prediction as it correlates with 

cccDNA in HBeAg-negative patients. Methods. We studied 

133 HBeAg-negative CHB patients treated with 48 weeks of 

peginterferon alfa-2a +/- ribavirin. Serum HBcrAg levels were 

measured at baseline and at week 12, and we assessed the 

correlation with sustained response (SR; ALT normalization 

& HBV DNA


was lower in patients with F4-F6 (n=27) compared to those 

with F0-F3 (7.6 vs. 8.1 log U/mL, p=0.02), and declined with 

increasing fibrosis stage (Figure). HBcrAg levels were independently 

associated with advanced fibrosis when adjusting 

for age and serum ALT (OR 0.5, CI-95% 0.3-0.8, p=0.003). 

In HBeAg-negative patients, fibrosis stage was available for 

all patients. The mean HBcrAg was 5.5 (SD 0.1), median HAI 

score 5 (IQR 3), and median Ishak fibrosis stage 3 (IQR 2). 

Genotypes A/B/C/D/other were present in 13/1/2/81/3%. 

In these patients, log HBcrAg was not correlated with Ishak 

fibrosis score (r s 

=-0.1, p=0.59), similar to fibrosis correlation 

to qHBsAg (r s 

=-0.1, p=0.19), and HBV DNA (r s 

=0.2, 0.07). 

When adjusting for age and serum ALT, HBcrAg levels were 

not associated with advanced fibrosis (OR 0.9, CI-95% 0.5- 

1.6, p=0.83) and did not change with increasing fibrosis 

stage (Figure). For both HBeAg-positive and HBeAg-negative 

patients, variation of qHBsAg levels across fibrosis stages 

was comparable to that of HBcrAg levels (Figure). Conclusion. 

In this study, serum HBcrAg levels were inversely correlated 

with Ishak fibrosis stage in HBeAg-positive CHB, but not in 

HBeAg-negative CHB. 

obtainable mutation analysis in 2 centers. Of 177 HBeAg(+) 

patients, 6(3.39%) were genotype A, 65(36.72%) genotype 

B, 104(58.76%) genotype C, 1(0.57%) genotype B/C, and 

1(0.57%) genotype C/D. Of 90 HBeAg(-) patients, 1(1.11%) 

were genotype A, 50(55.56%) genotype B, 37(41.11%) genotype 

C and 2(2.22%) genotype D. When compared to HBeAg(-) 

patients, HBeAg(+) patients were significantly younger in mean 

age (37.93 vs. 44.40; P


able on post-delivery flares in patients treated with tenofovir 

(TDF) during pregnancy. Aim: Cross-sectional observational, 

single centre study to compare the frequency of post-delivery 

flares in untreated CHB mothers and in CHB patients 

requiring therapy during pregnancy (due to liver disease or 

prevent HBV transmission) and evaluate the importance virological 

(HBV DNA), serological (HBeAg and HBsAg levels) 

and immunological markers (IP10 levels) during pregnancy 

(2 nd trimester) on predicting post-delivery flares. Patients: 297 

CHB patients (median age 31.1years, 16% HBeAg+) were 

assessed at 2 nd pregnancy trimester (1 st visit median gestation 

week 26); 35 patients were treated with TDF prior to pregnancy, 

39 patients started TDF from gestation week 28 to prevent 

HBV transmission and 223 untreated patients (all HBV 

DNA38IU/l and increase by two-fold pregnancy level. 36 

patients stopped therapy shortly after delivery. Results: Treated 

patients had higher frequency of flares than untreated mothers 

(51% vs. 26%,p200pg/ml) (OR 

4.6, 1.8-11.4), HBV DNA (>10000IU/ml) (OR 9.2, 4.9-16.9), 

HBsAg (>20000IU/ml) (OR 5.5, 2.5-12.2) and higher HBeAg 

(>1000S/CO) levels (OR 5.7, 0.5-60) at 2 nd pregnancy trimester. 

While only high IP10 was predictive of HF in untreated 

mothers (OR 4.2, 1.5-10.2); higher HBV DNA, HBsAg, HBeAg 

and IP10 levels and younger age ( Methods: Based on Taiwan National Health Insurance 

Research Database, 176,120 patients diagnosed with RA or 

psoriasis were screened for eligibility. Only those having past 

HBV infection were screened. After excluding those having 

other hepatitis, taking antiviral drugs for viral hepatitis or those 

with malignant diseases, we enrolled a total of 354 eligible 

patients who received TNF-α antagonists (biologics cohort) 

and matched them 1: 2 with 708 patients who received nbD- 

MARDs alone by baseline characteristics and propensity scores 

(DMARDs cohort). Both cohorts were followed up for the occurrence 

of HBV hepatitis flare after starting TNF-α antagonists. 

Results: The 5-year cumulative incidences of HBV hepatitis flare 

were 19.3% (95% confidence interval [CI] 13.8-24.9%) and 

12.9% (95% CI 9.2-16.6%) for the Biologics and DMARDs 

cohorts, respectively (P=0.004). On modified Cox proportional 

hazards analysis, patients in the biologics cohort had 

significantly higher risk of HBV hepatitis flare (adjusted HR 

2.10, 95% CI 1.3-3.3, P Conclusion: 

Biologics use is associated with significantly higher risk of 

HBV hepatitis flare than DMARDs in RA or psoriasis patients. 

Disclosures: 

The following authors have nothing to disclose: Chun-Ying Wu, Yi-Ju Chen, Jaw- 

Town Lin 

1590 

The PAGE-B Score Accurately Predicts Clinical Outcome 

and Outperforms Other Biomarkers over 15 Years of 

Follow-up in a Diverse Cohort of Chronic Hepatitis B 

Patients 

Willem Pieter Brouwer 1 , Adriaan J. van der Meer 1 , Andre Boonstra 

1 , Elisabeth P. Plompen 1 , Suzan D. Pas 1 , Robert J. de Knegt 1 , 

Robert A. de Man 1 , Fiebo J. ten Kate 1 , Harry L. Janssen 2,1 , Bettina 

E. Hansen 1 ; 1 Gastroenterology & Hepatology, Erasmus Medical 

Center Rotterdam, Rotterdam, Netherlands; 2 Gastroenterology and 

Hepatology, UHN Liver Clinic, Toronto, ON, Canada 

Background & aims. Multiple non-invasive markers have been 

associated with hepatocellular carcinoma (HCC) development 

in chronic hepatitis B (CHB) patients. We aimed to compare the 

prognostic performance of these markers and to assess whether 

liver histology could improve this performance. Methods. Liver 

biopsies from consecutive treatment-naïve CHB patients were 

scored by a single experienced hepato-pathologist. Laboratory 

values at the time of biopsy were used to calculate the PAGE-B, 

REACH-B, GAG-HCC, and CU-HCC scores. A clinical event 

was defined as a combined endpoint including HCC development, 

liver failure, liver transplantation, and all-cause mortality. 

Event data was obtained from national database registries. 

Results. Of 557 patients, mean age at biopsy was 35±13 

years, 47/31/19% was Caucasian/Asian/African, 63% 

received antiviral therapy (AVT) after liver biopsy and 113 

(20%) had advanced fibrosis (F3/F4). Forty patients developed 

a clinical event within a median follow-up of 10.1 years 

(IQR 5.7-15.9, complete follow-up 93%). The PAGE-B score 

predicted any clinical outcome (C-statistic [C] 0.87, 95%CI: 

0.81-0.92), HCC development (C: 0.89) and reduced transplant-free 

survival (C: 0.84) with good accuracy, also when 

stratified by ethnicity, AVT after biopsy or advanced fibrosis. 

The REACH-B, GAG-HCC, and CU-HCC predicted clinical outcome 

less accurate (C: 0.70, 0.82, and 0.73, respectively).


The event prediction with the PAGE-B improved modestly with 

the Ishak fibrosis score (C: 0.88), but the net reclassification 

improvement appeared not clinically significant. Conclusion. 

The PAGE-B score showed the best performance to assess the 

likelihood to develop a clinical event among a diverse CHB 

population over 15 years of follow-up. Additional liver histologic 

characteristics did not appear to improve this event risk 

assessment. 

27.6%, 12.1%, 4.0% and 1.4% for patients with LC aged ≥ 

50 years, those with LC aged < 50 years, those without LC 

aged ≥ 50 years, and those without LC aged < 50 years, 

respectively. HBV DNA levels and quantitative hepatitis B surface 

antigen (qHBsAg) levels were independent predictors for 

inactive carriers in patients without LC. The 5- year cumulative 

incidence of an inactive carrier was 67.9% in non-LC patients 

with both low qHBsAg and HBV DNA levels, while the disease 

progression rate was 2.2 %. Conclusions: HBeAg negative 

patients without LC can be monitored for becoming an inactive 

carrier when both HBV DNA levels and qHBsAg levels are 

low, as the risk of disease progression is low while incidence 

of inactive carrier is high. 

Cumulative incidence of inactive carriers according to HBV DNA 

and qHBsAg levels in patients without cirrhosis. A-D represent low 

HBV DNA/low qHBsAg, low HBV DNA/high qHBsAg, high HBV 

DNA/low qHBsAg and high HBV DNA/high qHBsAg, respectively. 

Disclosures: 


Gilead 


Roche, Gilead 











The following authors have nothing to disclose: Willem Pieter Brouwer, Elisabeth 

P. Plompen, Suzan D. Pas, Fiebo J. ten Kate, Bettina E. Hansen 

1591 

Prediction of Clinical Outcomes in Hepatitis B E Antigen 

Negative Chronic Hepatitis B Patients with Elevated 

Hepatitis B Virus DNA Levels 

Jem Ma Ahn, Dong Hyun Sinn, Geum-Youn Gwak, Yong Han 

Paik, Moon Seok Choi, Joon Hyeok Lee, Kwang Cheol Koh, Seung 

Woon Paik; Department of Medicine, Samsung Medical Center, 

Sungkyunkwan University School of Medicine, Seoul, Korea (the 

Republic of) 

We investigated whether long-term clinical outcomes such as 

disease progression or inactive hepatitis B virus (HBV) carrier 

can be predicted by baseline factors in hepatitis B e antigen 

(HBeAg)-negative HBV infected patients with elevated viral 

loads. A retrospective cohort of 527 HBeAg-negative chronic 

HBV infected patients with elevated viral loads (HBV DNA ≥ 

2,000 IU/ml) were assessed for disease progression defined 

by development of hepatocellular carcinoma or cirrhosis (LC) 

complications, as well as becoming an inactive carrier, defined 

by a decrease of HBV DNA < 2,000 IU/ml for ≥ 6 months in 

absence of antiviral therapy. During a median 3.6 years of 

follow-up, disease progression was detected in 46 patients, 

and 79 patients became inactive carriers. Old age and cirrhosis 

were independent predictors for disease progression. 

The 5-year cumulative incidence of disease progression were 

Disclosures: 

The following authors have nothing to disclose: Jem Ma Ahn, Dong Hyun Sinn, 

Geum-Youn Gwak, Yong Han Paik, Moon Seok Choi, Joon Hyeok Lee, Kwang 

Cheol Koh, Seung Woon Paik 

1592 

The Role of Surface Antibody in Hepatitis B Virus Reactivation 

in Patients with Resolved Infection: A Systematic 

Review and Meta-Analysis 

Sonali Paul 1 , Aaron Dickstein 1 , Akriti P. Saxena 1 , Norma Terrin 1 , 

Kathleen Viveiros 1 , Ethan M. Balk 2 , John B. Wong 1 ; 1 Tufts Medical 

Center, Cambridge, MA; 2 Brown University School of Public 

Health, Providence, RI 

Background: Patients with resolved hepatitis B virus (HBV) and 

hematological malignancies are at risk for HBV reactivation. 

Data about whether hepatitis B surface antibody (HBsAb+) 

reduces the risk of reactivation are conflicting and limited. 

This systematic review with meta-analysis assesses whether the 

presence of HBsAb+ reduces the risk of HBV reactivation in 

patients with resolved HBV infection (negative surface antigen, 

HBsAg-, and positive core antibody, HBcAb+) receiving 

chemotherapy for hematological malignancies. Methods: We 

included English language studies through March 31, 2015 

that examined reactivation in patients with resolved HBV 

infection receiving chemotherapy for all hematological malignancies 

from MEDLINE, Web of Science, Cochrane Central 

Register of Controlled Trials, TOXNET, and Scopus. The pri-


mary outcome was HBV reactivation defined as an increase 

in HBV DNA levels from baseline or re-emergence of HBsAg+ 

when previously negative. The odds ratio (OR) of reactivation 

with versus without HBsAb+ was estimated with random-effects 

model meta-analyses. Results: Fourteen observational studies 

with 836 patients (range 8-354 with 7 patients receiving antiviral 

prophylaxis) met inclusion criteria. There were 9 prospective 

and 5 retrospective studies; 11 were from Asia and 3 from 

Italy; 10 included only lymphoma and 4 had mixed hematologic 

malignancies including leukemia and multiple myeloma. 

When reported, the median patient age across studies was 62 

years (range 18-90) with 334 males and 261 females. Chemotherapy 

regimens varied across studies with rituximab in 8 

studies. The overall pooled OR was 0.20 (95% CI 0.11–0.35; 

with no heterogeneity, I 2 =0%) indicating a protective effect 

of HBsAb+ on the risk of reactivation versus HBcAb+ alone. 

The absolute risk of reactivation with HBcAb+ alone was 17% 

(95% CI 10-25%). Similar results were found when the analysis 

was limited to rituximab-based chemotherapy (OR 0.17, 95% 

CI 0.08–0.34) and lymphoma (OR 0.17, 95% CI 0.09–0.32). 

Sensitivity analysis found a significant protective effect of 

HBsAb+ in 9 prospective studies but not in 5 retrospective studies. 

The ORs continued to remain significant when analyzed by 

geographic region (Asia OR 0.22, 95% CI 0.12–0.40; Italy 

OR 0.07, 95% CI 0.01–0.54). Conclusions: In patients with 

resolved HBV receiving chemotherapy for hematologic tumors, 

HBsAb+ decreases the risk of reactivation. Our results support 

the need for future studies examining the effect of HBsAb titers 

and booster vaccinations prior to chemotherapy in this patient 

population. 

Disclosures: 

The following authors have nothing to disclose: Sonali Paul, Aaron Dickstein, 

Akriti P. Saxena, Norma Terrin, Kathleen Viveiros, Ethan M. Balk, John B. Wong 

and the death certification system was performed. Cox proportional 

hazards models were used to estimate the multivariate-adjusted 

hazard ratio of developing LC and HCC. Results: 

Incidence rates of LC per 100,000 person-years were 341.2, 

565.5, 654.1, 920.0, 1477.9 for serum anti-HBc levels of 


D. At baseline, serum levels of HBV DNA, HBsAg and HBV 

RNA did not differ in patients with and without subsequent 

HBeAg SC. On treatment, HBV DNA and HBsAg levels did not 

differ between patients with and without HBeAg SC (Table). 

However, HBeAg positive patients achieving HBeAg SC had 

rapid HBV RNA declines by week 12 with most below LLOD by 

week 24. In contrast, HBeAg positive patients without SC had 

significantly higher levels of HBV RNA at all time points tested 

during treatment (p


tive samples. Out of which 280 (48.4%) samples were found 

to be HBV DNA positive and genotyping was done for 228 

samples. The frequency distribution of genotypes were genotype 

D=214 (93.8%), genotype C=12 ( 5.2%), Genotype 

I=2(0.8%). Genotype D (93.8%) was found to be predominant 

genotype followed by C (5.2%) in the North eastern states. 

Conclusion:- HBV prevalence rate in North eastern states was 

found to be 5.9%. Genotype D (93.8%) was found to be predominant 

genotype followed by C (5.2%) in the North eastern 

states. This analysis of population-based data provides 

evidence of the prevalence of HBV infection in North eastern 

states. This important finding adds to the existing data of HBV 

prevalence in India. These results are relevant to public health 

policy makers and highlight the importance of promoting hepatitis 

B vaccination programs in the endemic areas of high HBV 

prevalence. 

Disclosures: 

The following authors have nothing to disclose: Premashis Kar, Vijay Karra 

1597 

Performance of the new cobas ® HBV Assay on the 

cobas ® 6800 system in comparison to different 

TaqMan ® assays 

Benjamin Maasoumy 1 , Kevin Luk 2 , Birgit Bremer 1 , Patrick Lehmann 

1 , Ed G. Marins 2 , Veronique Michel-Treil 3 , Ekaterina Gelman 

3 , Merlin Njoya 2 , Jesse A. Canchola 2 , Christian O. Simon 4 , 

Heiner Wedemeyer 1 ; 1 Gastroenterology, Hepatology and Endocrinology, 

Hannover Medical School, Hannover, Germany; 2 Roche 

Molecular Systems Inc., Roche Diagnostics, Pleasanton, CA; 

3 Covance Central Laboratory Services SA, Geneva, Switzerland; 

4 Roche Diagnostics International, Ltd, Rotkreuz, Switzerland 

Background: Management of patients with hepatitis B (HBV) 

critically relies on robust and sensitive HBV DNA assays. Various 

commercial HBV DNA assays that are based on amplifying 

and detecting HBV DNA have received CE Mark or FDA 

Approval. In 2014, the new cobas ® 6800/8800 systems were 

introduced and are now used in routine diagnostic practice. 

For this platform, a new HBV viral load test was developed. 

We here aimed to investigate the performance of this new 

HBV DNA assay in a routine diagnostic setting. Methods: We 

evaluated the performance of the new cobas ® HBV test on the 

cobas ® 6800 system in comparison to the version 2.0 of the 

COBAS ® AmpliPrep/ COBAS ® TaqMan ® HBV test with automated 

nucleic acid extraction (HBV TaqMan ® v2) and to the 

manual extraction COBAS ® TaqMan ® HBV test For Use With 

The High Pure System (HBV HPS). Up to 336 HBV WHO 3 rd 

International Standard dilutions as well as 285 clinical samples 

were tested each with all the three assays. Results: The lower 

limits of detection were 2.7 IU/ml, 9.6 IU/ml and 6.2 IU/ 

ml for cobas ® HBV, HBV TaqMan ® v2 and HBV HPS assays, 

respectively, based on up to 48 samples tested for each dilution 

(2000 IU/mL, 200 IU/mL, 60 IU/ml, 20 IU/mL, 10 IU/ 

mL, 5 IU/mL, and 2.5 IU/mL). Cobas ® HBV detected 48/48 

samples with a nominal titer of 5 IU/mL and 40/48 samples 

with a titer of 2.5 IU/mL. The assay was highly reproducible 

and linear even in samples with very low viremia with an average 

log10 IU/mL difference between observed and nominal 

titers of 0.04 (respective values were 0.05 and 0.18 for the 

HBV TaqMan ® v2 and the HBV HPS assays). Parallel testing 

of clinical samples showed largely comparable values across 

different viral loads with minimally lower values revealed by 

cobas ® HBV compared to HBV TaqMan ® v2 (mean of paired 

difference -0.099 log10 IU/mL; 95% CI -0.132; -0.065), while 

no significant difference was observed compared to the HBV 

HPS (mean of paired difference -0.011 log10 IU/mL; 95% CI 

-0.044; +0.023). Classification of samples at critical treatment 

decision cut-offs showed an overall percentage of agreement 

between the cobas ® HBV and the HBV TaqMan ® v2 assays 

of 94.4% for both the 2000 IU/mL and 20000 IU/mL thresholds 

while the respective values were 95.4% and 97.3% for 

cobas ® HBV vs. HBV HPS. Conclusions: The new cobas ® HBV 

test demonstrated an improved sensitivity compared to current 

TaqMan ® HBV assays, however, the clinical meaning of very 

low HBV viremia detected with new assay needs to be determined. 

Quantitative HBV DNA values are highly comparable 

between the assays, which demonstrates that the cobas ® HBV 

test can be reliably used in clinical practice to guide treatment 

decisions. 

Disclosures: 

Benjamin Maasoumy - Advisory Committees or Review Panels: Abbott Molecular, 

Janssen-Cilaq; Grant/Research Support: Abbott Molecular; Speaking and Teaching: 

MSD, Roche Diagnostics, Roche Pharma, Janssen-Cilaq, Fujirebio, BMS 

Ed G. Marins - Employment: Roche Molecular Systems 

Merlin Njoya - Employment: Roche 

Jesse A. Canchola - Employment: Roche Molecular Diagnostics Inc. 

Christian O. Simon - Employment: Roche Diagnostics International 





The following authors have nothing to disclose: Kevin Luk, Birgit Bremer, Patrick 

Lehmann, Veronique Michel-Treil, Ekaterina Gelman 

1598 

Seroprevalence and Clinical Features of Hepatitis D 

Virus (HDV) Infection in a North American Cohort 

Norah Terrault 1 , Marc G. Ghany 2 , Chaeryon Kang 3 , Stewart 

Cooper 4 , Adrian M. Di Bisceglie 5 , Michael W. Fried 6 , Steven 

H. Belle 3 , Jay H. Hoofnagle 2 ; 1 UCSF, San Francisco, CA; 2 NIH- 

NIDDK, Bethesda, MD; 3 University of Pittsburgh, Pittsburgh, PA; 

4 California Pacific Medical Center, San Francisco, CA; 5 St. Louis 

University, St. Louis, MO; 6 University of North Carolina, Chapel 

Hill, NC 

Background Hepatitis D virus (HDV) infection is estimated to 

affect ~4-5% (15 million) of persons with chronic hepatitis B 

virus (HBV) worldwide. The burden and clinical characteristics 

of HDV infection in North America is unknown. Aim: To determine 

the prevalence and clinical features of HDV in a well-characterized 

cohort of adults in North America with chronic HBV. 

Methods: All HBsAg-positive adults in the HBRN cohort with 

serum samples at enrollment were tested for total anti-HDV 

using a commercially available assay (DiaSorin, Italy). Prevalence 

of HDV was examined with respect to demographic and 

clinical features at enrollment. Results: Of 1165 adults tested, 

30 (2.6%) were anti-HDV positive and 4 (0.3%) equivocal. For 

analysis of clinical features, 14 with HCV (13 HDV-negative; 

one triple-infected HBV/HCV/HDV) were excluded. Prevalence 

of anti-HDV positivity was similar in men (2.3%) and women 

(2.7%) and among whites (3.2%), blacks (4.9%) and Asians 

(1.8%) (p=0.08). Prevalence of HDV did not differ significantly 

by age (median 42 vs. 50 yrs) or presumed route of infection. 

Prevalence differed by genotype (p=0.001) being highest in 

participants with genotypes D (10.7%) and E (8.0%) which 

was also reflected in the differing prevalence of HDV by region 

of birth (p


ALT or AST levels, platelet counts, HBsAg levels, or HBeAg 

prevalence were similar between HDV positive and negative 

participants. Conclusions: HDV coinfection was infrequent 

(~3%) among adults in this North American HBV cohort. A distinctive 

risk profile for acquisition was not apparent, highlighting 

the challenges in using a risk-based screening algorithm to 

identify HDV-infected adults. 

Laboratory/Viral Features 

*median (range) 

Disclosures: 









Steven H. Belle - Grant/Research Support: Rottapharm!Madaus 

The following authors have nothing to disclose: Marc G. Ghany, Chaeryon Kang, 

Stewart Cooper, Jay H. Hoofnagle 

1599 

Comparative Study for Anti-Hepatitis B Surface Antigen 

Titers Based on Two Measurement Methods: Using 

Monoclonal Antibodies Isolated from Hepatitis B Vaccinated 

Japanese Recipients. 

Takako Inoue 1 , Shuko Murakami 2 , Susumu Tsutsumi 2 , Kumiko 

Ohne 1 , Kazuto Tajiri 3 , Hiroyuki Kishi 4 , Shintaro Ogawa 2 , Noboru 

Shinkai 5 , Takaaki Goto 1 , Yukio Wakimoto 1 , Yasuhito Tanaka 2,1 ; 

1 Clinical Laboratory, Nagoya City University Hospital, Nagoya, 

Japan; 2 Department of Virology & Liver unit, Nagoya City University 

Graduate School of Medical Sciences, Nagoya, Japan; 

3 Department of Immunology, University of Toyama, Toyama, Japan; 

4 Department of Immunology, Graduate School of Medicine and 

Pharmaceutical Sciences, University of Toyama, Toyama, Japan; 

5 Department of Gastroenterology and Metabolism, Nagoya City 


Background: Since anti-hepatitis B surface antigen (anti-HBs) 

titers are various depending on measurement methods, we 

compared two different methods to quantity anti-HBs titers in 

sera and HBs monoclonal antibodies. Methods: 1) The measurement 

of anti-HBs was compared using either Lumipulse 

G1200 (Fujirebio Inc.) or Architect i2000SR (Abbott Japan). 

Previously, we isolated human monoclonal antibodies (mAbs) 

against HBV from Japanese healthy volunteers who had been 

immunized with a genotype C recombinant HBV vaccine 

(Biimugen, Kaketsuken), using a cell-microarray system. A following 

report revealed that among these mAbs, HB0116 and 

HB0478, recognize the first N-terminal peptide loop within 

the “a” determinant and have HBV-neutralizing activities. In 

this study, HB0116, HB0478 and four other mAbs were used. 

2) The sera from 182 HBV-resolved patients in our hospital, 

who were negative for hepatitis B surface antigen (HBsAg) 

but positive for anti-hepatitis B core antigen (anti-HBc) and/or 

anti-HBs, were obtained. This study protocol was approved by 

the appropriate institutional ethics review committees. Results: 

1) Measuring 2 mAbs (HB0116 and HB0478) with HBV-neutralizing 

activities, the anti-HBs titers of HB0116 (1.0 μg/mL) 

were comparable (440.7 mIU/mL by Architect and 689.3 

mIU/mL by Lumipulse), whereas those of HB0478 (1.0 mg/ 

mL) were much lower by Architect than by Lumipulse (42.6 

mIU/mL by Architect and 818.6 mIU/mL by Lumipulse). Of 

the other 4 mAbs without HBV-neutralizing activities, equal 

titers were observed for one; two mAbs had less anti-HBs titers 

by Architect; and one was below the cut-off index (


(95%CI 1.01 − 1.09), p=0.0018), AFP > 20 ng/mL (HR 3.2 

(95%CI 1.01 − 9.99), p=0.049), WFA(+)-M2BP (per 1 COI) 

(HR 1.3 (95%CI 1.01 – 1.63), p=0.039) as independent risk 

factors for the development of HCC. Conclusion: WFA(+)-M2BP 

can be applied as a simple and reliable surrogate marker for 

the risk of HCC development, in addition to liver fibrosis stage 

as determined by liver biopsy. 

Disclosures: 

Seigo Abiru - Grant/Research Support: CHUGAI PHARMACEUTICAL CO.,LTD 

The following authors have nothing to disclose: Kazumi Yamasaki, Shigemune 

Bekki, Yuki Kugiyama, Shinjiro Uchida, Akira Saeki, Satoru Hashimoto, Shinya 

Nagaoka, Atsumasa Komori, Hiroshi Yatsuhashi 

1601 

Identification of a non-invasive model based on serum 

levels of miR-122 and miR-222 to predict severe fibrosis 

and cirrhosis in patients with chronic hepatitis B 

Kevin Appourchaux 5,2 , Emilie Estrabaud 5,2 , Matthieu Resche-Rigon 

3,4 , Martine Lapalus 5,2 , Michelle Martinot-Peignoux 5,2 , Nathalie 

Boyer 2 , Michel Vidaud 6 , Pierre Bedossa 7,1 , Patrick Marcellin 5,2 , 

Tarik Asselah 5,2 ; 1 INSERM, Paris, France; 2 Hepatology, Beaujon 

Hospital, Clichy, France; 3 Service de Biostatistique et information 

médicale, Saint-Louis Hospital, Paris, France; 4 INSERM UMR1153, 

Paris, France; 5 INSERM UMR1149, Paris, France; 6 INSERM 

UMR745, Paris, France; 7 Service d’Anatomie Pathologique, Beaujon 

Hospital, Clichy, France 

Background and aims Patients with chronic hepatitis B (CHB) 

are at high risk to develop cirrhosis and hepatocellular carcinoma 

(HCC). Staging fibrosis is mandatory, for both the prognosis 

and the need for treatment. The liver-enriched miR-122 

has been suggested to inhibit HBV replication. MicroRNAs are 

increasingly investigated as biomarkers because of their high 

stability. We aimed to identify miRNAs differentially expressed 

during fibrosis in patients with CHB. Patients and Methods A 

total of 103 patients with CHB were consecutively enrolled. All 

the patients had at least one biopsy to determine the stage of 

fibrosis (METAVIR scoring system) and no HCC. 85,4% of the 

patients were males, the mean age was 41.8 years. The mean 

HBV DNA level was 5.74 logUI/mL and 60.2% of the patients 

were negative for HBe antigen. Serums and biopsies were 

available for respectively 88 and 83 patients (69 paired liver/ 

serum). Among patients with available serums 2.3%, 26.4%, 

27.6%, 24.1% and 19.6% had respectively F0, F1, F2, F3 

and F4 ; and among patients with available biopsies, 2.4%, 

27.7%, 27.7%, 20.5% and 21.7% had respectively F0, F1, 

F2, F3 and F4. The expression of miR-27b, -29a, -92a, -122, 

-146a, -222 and -224 which are related to liver fibrosis was 

assessed by RT-qPCR. Results A reduced expression of hepatic 

miR-122 (p=0.004) and miR-27b (p=0.038) was observed in 

patients with severe fibrosis and cirrhosis (F3-F4) compared 

to those with no, mild and moderate fibrosis (F0-F1-F2). An 

increased expression of hepatic miR-222 (p=0,018) and 

miR-224 (p=0.0001) was observed in patients with F3-F4 

compared to those with F0-F1-F2. A reduced expression of 

circulating miR-122 (p=0.049), miR-92a (p=0.031), and miR- 

29a (p=0.048) was observed in patients with F3-F4 compared 

to those with F0-F1-F2. An increased expression of circulating 

miR-146a (p=0.015) and miR-222 (p=0.040) was observed 

in patients with F3-F4 compared to those with F0-F1-F2. The 

multivariate analysis of clinical data and circulating miRNAs, 

allowed us to build a model combining circulating miR-122 

and miR-222, platelets count and alkaline phosphatase (ALP). 

The AUC of the model was 0,86 while FIB-4 and APRI had 

an AUC of 0,81 and 0,70, respectively. Conclusions Hepatic 

miR-27b, -122, -222 and -224 were differentially expressed in 

patients with F0-F1-F2 and in those with F3-F4. The increased 

expression of miR-222 in patients with F3-F4 might suggest 

a premalignant condition to HCC. The model combining the 

assessment of the following non-invasive biomarkers, circulating 

miR-122 and miR-222, platelets count and ALP was more 

accurate than FIB-4 and APRI to distinguish patients with F3-F4 

from those with F0-F1-F2. 

Disclosures: 







Abbvie 



The following authors have nothing to disclose: Kevin Appourchaux, Emilie 

Estrabaud, Matthieu Resche-Rigon, Martine Lapalus, Michelle Martinot-Peignoux, 

Michel Vidaud, Pierre Bedossa 

1602 

Interleukin-2 Receptor and Transforming Growth Factor 

Alpha Independently Predict Significant Fibrosis in 

Chronic Hepatitis B Patients with Normal or Mildly Elevated 

Alanine Aminotransferase 

Gui-Qiang Wang, Yong-Qiong Deng, Hong Zhao; Department of 

infectious disease, Peking University First Hospital, Beijing, China 

Background: Patients with normal or mildly elevated serum alanine 

aminotransferase (ALT) are not guaranteed to be free from 

liver damage. Invasive liver biopsy is the current gold standard 

for assessing histological liver injury. There is an increasing 

demand for developing a noninvasive marker or index for diagnosis 

of liver inflammation and fibrosis in patients with normal 

or mildly elevated ALT.Aim: The aim of this study was to investigate 

the association of circulating cytokines and chemokines 

with liver inflammation and fibrosis, and to deeply assess their 

diagnostic value in CHB patients with ALT less than 2 times of 

upper limit of normal(ULN).Methods: Two hundred and twenty 

seven CHB patients with qualified biopsy were prospectively 

enrolled. There were 151 patients with ALT≥2×ULN and 76 

persons with ALT 2×ULN. All patients underwent liver biopsy. 

The serum levels of cytokines and chemokines were determined 

by simultaneous multianalyte detection methods. Histological 

Activity Index (HAI) and Liver fibrosis stage were assessed 

according to Ishak criteria.Results: Patients with moderate and 

more inflammation showed significantly higher levels of CXCL- 

11, CXCL-10 and interlerkin-2 receptor(IL-2 R)than the opposite 

(P0.001). Patients with significant fibrosis had higher levels of 

interleukin-8(IL-8) (P=0.027), Transforming growth factor alpha 

(TGF-a) (P=0.011), IL-2R (P=0.002) and CXCL-11(P=0.032) 

than no significant fibrosis. In 151 patients with ALT≤2×ULN, 

31.8% and 29.1% showed moderate and more inflammation 

and significant fibrosis respectively. Multivariate analysis indicated 

CXCL-11 independently associated with moderate and 

more inflammation, and TGF-a, L-2R with significant fibrosis 

in patients with ALT≤2×ULN. Based on CXCL-11, TGF-a, L-2R 

and clinical parameters, we developed inflammation-index 

and fib-index which showed areas under the receiver operating 

characteristics curve (AUROC) of 0.75 (95% CI 0.66- 

0.84) for moderate and more inflammation, and 0.82(95% CI 

0.75, 0.90) for significant fibrosis respectively in patients with 

ALT≤2×ULN. Leave-one out cross-validation showed 74.5% 

and 81.3% cases correctly classified by inflammation-index 

and fib-index. Compared to existing scores, fib-index was 

significantly superior to aspartate aminotransferase (AST) to 

platelet ratio index (APRI) and FIB-4 score for significant fibro-


sis.Conclusion: IL-2R and TGF-a were independent predictors 

for significant fibrosis in CHB patients with normal or mildly 

elevated ALT. An IL-2R and TGF-a based score, fib-index, was 

superior to APRI, FIB-4 for the diagnosis of significant fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Gui-Qiang Wang, Yong-Qiong 

Deng, Hong Zhao 

1603 

Characterization of Nucleic acid testing reaction samples 

and Occult Hepatitis B infection Among blood 

donors in three cities of China 

Xiaomei Wang, Xiumei Chi, Ruihong Wu, Xiuzhu Gao, Junqi 

Niu; Hepatology, The First Hospital of Jilin University, Changchun, 

China 

Introduction. In China, HBV infection is common. With increasing 

voluntary blood donation and the still-prevalent infectious 

diseases in donors, we need to augment transfusion-transmitted 

infections testing before use. Our study was aimed to compare 

the seroprevalence of HBV, HCV and HIV among the 

donors of Chineses tested by ELISA and nucleic acid testing 

(NAT) . A variant of hepatitis B virus (HBV) having a specific 

mutation within the S gene has been found in occult HBV infection 

(OBI) . To know whether similar variants were involved in 

blood donors, we analyzed 18 blood samples HBV S region 

sequence. Materials and Methods. Enzyme linked immunosorbent 

assay (ELISA) was used for detection of HBsAg, HBeAg, 

anti-HBs, anti-HBe, anti-HBc,anti-HIV, and anti-HCV in all donor 

serum. The blood samples which were negative on ELISA were 

also subjected to NAT testing for HBV, HCV, and HIV. Nested 

PCR were used to amplify the S regions and products were 

sequenced and analyzed. Results. A total of 482370 donations 

were tested in studied, NAT yielded 156 HBV-DNA-positive 

donations in the HBsAg-negative (1:2365). NAT yielded 2 

HIV-RNA-positive donations were p24 Ag reactive. There were 

no NAT positive in anti-HCV negative donations. There were 

no escape mutations observed in the S gene. Conclusion. In 

regions with a high prevalence of HBV,there are likely to be 

a significant number of OBI donations that can be identified 

by NAT. Therefor, in order to provide safe blood, NAT should 

be applied in blood centers for testing HBV.In 18 occult HBV 

infections, HBV sequences lacked G145R and other escape 

mutations in S region, which may be caused by wild-type HBV 

strains. 

Disclosures: 

The following authors have nothing to disclose: Xiaomei Wang, Xiumei Chi, 

Ruihong Wu, Xiuzhu Gao, Junqi Niu 

1604 

Baseline quantitative hepatitis B core antibodies can 

predict HBV DNA and HBsAg seroclearance in treatment-naïve 

HBeAg negative chronic hepatitis B patients 

Hui-Han Hu 1 , Jessica Liu 1 , Chia-Lin Chang 1 , Chin-Lan Jen 1 , Mei- 

Hsuan Lee 2 , Sheng-Nan Lu 3 , Li-Yu Wang 4 , San-Lin You 1 , Pei-Jer 

Chen 5,6 , Hwai-I Yang 1 , Chien-Jen Chen 1,7 ; 1 Genomics Research 

Center, Academia Sinica, Taipei, Taiwan; 2 Institute of Clinical 

Medicine, National Yang-Ming University, Taipei, Taiwan; 

3 Department of Gastroenterology, Chang-Gung Memorial Hospital, 

Kaohsiung, Taiwan; 4 MacKay College of Medicine, Taipei, 

Taiwan; 5 Division of Gastroenterology, Department of Internal 

Medicine, National Taiwan University Hospital, Taipei, Taiwan; 

6 National Taiwan University, Graduate Institute of Clinical Medicine, 

Taipei, Taiwan; 7 Graduate Institute of Epidemiology and 

Preventative Medicine, College of Public Health, National Taiwan 

University, Taipei, Taiwan 

Hepatitis B core antibody (anti-HBc) is one of the classical seromarkers 

for HBV infection. Several lines of evidence showed 

that among chronic hepatitis B (CHB) patients, quantitative 

anti-HBc levels were strongly correlated with serum ALT levels, 

suggesting that anti-HBc is a surrogate indicator of immune 

activation. In addition, among treated HBeAg-seropositive CHB 

patients, baseline quantitative anti-HBc levels were strong predictors 

for treatment responses including HBeAg seroconversion. 

Here, we investigated the impact of baseline quantitative 

anti-HBc levels on spontaneous HBV DNA and HBsAg seroclearance 

in treatment-naïve HBeAg negative CHB patients 

from the REVEAL-HBV cohort (n=2503). Baseline anti-HBc levels 

were assessed using a newly developed double-sandwich 

anti-HBc immunoassay. HBV DNA and HBsAg levels were longitudinally 

evaluated with repeated measurement in the longterm 

follow-up samples. The associations of baseline anti-HBc 

levels with HBV DNA and HBsAg seroclearance were assessed 

by multivariate-adjusted Cox proportional hazard regression 

models. The rate ratios (RR adj 

) with 95% confidence intervals 

(CI) were estimated. Baseline HBV DNA levels were significantly 

and positively associated with increasing anti-HBc levels. 

Mean log 10 

(anti-HBc) levels were 2.82, 3.64, 3.95, 4.14, 

and 4.29, respectively, for patients with HBV DNA levels of 


The following authors have nothing to disclose: Hui-Han Hu, Jessica Liu, Chia-Lin 

Chang, Chin-Lan Jen, Mei-Hsuan Lee, Sheng-Nan Lu, Li-Yu Wang, San-Lin You, 

Hwai-I Yang, Chien-Jen Chen 

1605 

Risk assessment of hepatocellular carcinoma development 

using transient elastography vs. liver biopsy in 

chronic hepatitis B patients starting antiviral therapy 

Mi Na Kim 1 , Yeon Seok Seo 2 , Seung Up Kim 3 , Sang Gyune Kim 4 , 

Soon Ho Um 2 , Kwang-Hyub Han 3 , Young Seok Kim 4 ; 1 Department 

of Internal Medicine, CHA Gangnam Medical Center, CHA 

University, Seoul, Korea (the Republic of); 2 Department of Internal 

Medicine, Korea University College of Medicine, Seoul, Korea (the 

Republic of); 3 Department of Internal Medicine, Yonsei University 


of Internal Medicine, Soonchunhyang University Bucheon Hospital, 

Bucheon, Korea (the Republic of) 

Background/Aims: Liver stiffness (LS) assessed using transient 

elastography (TE) can assess the risk of developing hepatocellular 

carcinoma (HCC) in patients with chronic hepatitis B (CHB). 

We evaluated whether TE, when compared with histological 

data as a reference standard, can predict the risk of HCC 

development in CHB patients starting antiviral therapy. Methods: 

Database of 381 patients with CHB who underwent liver 

biopsy (LB) and TE starting antiviral therapy were reviewed. All 

patients underwent surveillance for HCC development using 

ultrasonography and alpha-fetoprotein. Results: During the 

median follow-up period of 48.1 (range 6.0-109.8) months, 

HCC developed in 34 (8.9%) patients. In patients with HCC 

development, age, proportion of diabetes mellitus, histological 

fibrosis stage, and LS value were significantly higher than those 

in patients without (all P


age-platelet index (API) for significant fibrosis (Metavir F2-4) 

in low-replicative (HBV DNA 4, 94.8 vs. 93.8%). At ROC-defined thresholds, APRI 

(≥0.33), AAR (≥0.93), FIB-4 (≥0.70) and API (>2) showed 

greater AUROCs for F2-4 diagnosis in low replicative (0.80, 

0.62, 0.81 and 0.71, respectively) vs. high-replicative patients 

(0.73, 0.52, 0.67 and 0.69, respectively). Conclusion: All 4 

biomarkers in both, low and high-replicative HBV demonstrate 

modest accuracy for fibrosis diagnosis at conventional cut-offs. 

Lowering the cut-offs may increase the diagnostic relevance of 

these biomarkers, particularly for APRI and FIB-4 in low-replicative 

disease. 

Disclosures: 

Faisal M. Sanai - Advisory Committees or Review Panels: Merck Sharpe Dohme, 

Bristol Myers Squibb, Janssen Pharmaceuticals, Gilead Sciences; Grant/ 

Research Support: Roche Pharmaceuticals, Bristol Myers Squibb; Speaking and 

Teaching: Roche Pharmaceuticals, Janssen Pharmaceuticals, Gilead Sciences, 

Bayer Schering 

Abdulrahman A. Aljumah - Advisory Committees or Review Panels: Gilead; 

Speaking and Teaching: Bristol-Myers Squibb 

Ibrahim H. Altraif - Consulting: MSD, ABBVIE, BRISTOL MYERS; Grant/Research 

Support: ROCHE, JANSSEN; Speaking and Teaching: GSK 

The following authors have nothing to disclose: Taha Farah, Khalid Albeladi, 

Faisal Batwa, Yaser Dahlan, Mohammed A. Babatin, Hamad I. Al-ashgar, 

Hadeel Al-mana, Khaled Alsaad, Khalid A. Alswat, Khalid I. Bzeizi 

1608 

Barriers to obtaining appropriate Hepatitis B care 

among US Veterans 

Tatyana Kushner, David E. Kaplan, Marina Serper; Division of 

Gastroenterology and Hepatology, Hospital of the University of 

Pennsylvania, Philadelphia, PA 

Background: Studies have demonstrated poor follow-up and 

adherence to guidelines in chronic hepatitis B (HBV) care. We 

sought to identify patient and health system-specific risk factors 

for gaps in quality of HBV care within the VA. Methods: 

We performed a retrospective analysis of 125 US Veterans 

with chronic HBV within 4 VA sites in Pennsylvania and Delaware 

from 1999-2012. Patients with acute HBV were excluded 

(n=12). Demographic and clinical data were obtained from 

medical records. Process of care outcomes were: specialist consultation, 

appropriate lab testing, and liver imaging. Results: 

The patients were 98% male, 50% African American, with a 

mean age of 51 (SD=13). A total of 18% were disabled and 

72% had ≥1 psychiatric comorbidity (depression, anxiety, or 

PTSD), 52% had ≥1 psychiatric hospitalization within 2 years 

of HBSAg+ result. A total of 8% reported marijuana use, 10% 

IV drug use, and 39% alcohol use; 28% of new cases were 

diagnosed as inpatients. A total of 7% had cirrhosis and 6% 

had hepatic decompensation. The median ALT was 38 (IQR 

25, 68). A total of 85% of subjects had primary care follow-up, 

with a mean 4 primary care visits per year; 16% were seen 

by infectious disease (ID) and 41% by gastroenterology (GI) 

specialists. The median time to specialist consult was 39 days 

(IQR: 4 to 255), 71% of patients did not have a consult ordered 

within 12 months. The median time to HBV follow up testing 

was 21 months (IQR 10 to 47); 33% of patients had complete 

laboratory testing with HBV DNA, HBeAg, and HBeAb 

testing. The median time to liver imaging was 4 months (IQR 

0.5 months to 14); 36% had confirmatory imaging within an 

ultrasound within 12 months. In bivariate analyses, psychiatric 

hospitalization was associated lower likelihood of specialist 

consultation (OR 0.75, 95% CI 0.56-0.98, p=0.03). African 

American race was associated with less confirmatory testing 

(OR=0.25 95% CI, 0.09-0.77, p=0.02). Receiving care at an 

outpatient vs. hospital VA facility was associated with lower 

rates of liver imaging (OR 3.5, 95% CI 1.2-10.2, p=0.02) and 

associated with lower receipt of either specialist consultation, 

appropriate lab testing, or liver imaging (OR 0.29, 95% CI 

0.09-0.94, p=0.04). Conclusion: Veterans with chronic HBV 

are significantly affected by psychiatric comorbidity and substance 

abuse. Delays in timely specialist referral, appropriate 

lab testing, and imaging were noted among certain subgroups 

(psychiatric comorbidity, African American race, outpatient VA 

facility). Prospective studies should confirm these relationships 

so that interventions to educate providers and improve care 

among high risk groups can be appropriately targeted. 

Disclosures: 

David E. Kaplan - Grant/Research Support: Bayer Pharmaceuticals, Inovio Pharmaceuticals 

The following authors have nothing to disclose: Tatyana Kushner, Marina Serper 

1609 

The Usefulness of CLIF-SOFA Score for Diagnosis and 

Evaluating the Patients with HBV-related Acute-on- 

Chronic Liver Failure 

Zhihong Wan, Chen Li, Shaoli You, HongLing Liu, ShaoJie Xin, 

Bing Zhu; Liver Failure Treatment and Research Center, 302 hospital 

of PLA, Beijing, China 

Background and Aims: Acute-on-chronic liver failure (ACLF) 

is a unique clinical entity. The definitions of ACLF used differed 

between Eastern and Western countries. Recently, a 

large prospective observational study (CANONIC study) was 

performed by EASL Chronic Liver Failure (EASL-CLIF) consortium. 

However, most of the enrolled patients were alcoholic 

cirrhosis, the criteria are necessary to be validated in China, 

in which the majority of ACLF is caused by chronic hepatitis B 

infection. In this regard, our present study was aimed to investigate 

clinical characteristics and prognosis in patients of HBV- 

ACLF diagnosed by EASL-CLIF criteria. Methods: Consecutive 

patients with HBV-ACLF were enrolled. All of the patients met 

the Chinese diagnostic criteria. EASL-CLIF criteria were used to 

further identify the patients with HBV-ACLF (HBV-ACLF-EASL). 

CLIF-SOFA score was used to assess organ failure and define 

ACLF grades. The mortality of the patients was recorded at 

28 days. Results: Total of 167 patients with HBV-ACLF was 

enrolled. Among them, 139 (83.2%) had liver cirrhosis and 

133 (79.6%) had acute decompensation of cirrhosis (AD). The 

28-day mortality rate was 24.6% in these patients. Among 

167 patients, thirty-six (21.6%) met the EASL-CLIF diagnostic 

criteria (diagnosed as HBV-ACLF-EASL). The 28-day mortality 

rate of these patients was 61.1%, which was significant higher 

than those with HBV-ACLF (24.6%, P < 0.01), indicating that 

patients with HBV-ACLF-EASL had more severity of disease. 

Of patients with HBV-ACLF-EASL, nine (25%) had ACLF grade 

1, 24 (66.7%) had ACLF grade 2, and 3 (8.3%) had ACLF 

grade 3. The most frequent organ failures were liver (97.2%)


and coagulation failures (58.3%) followed by cerebral (13.9%) 

and kidney failures (11.1%). The mortality was 44.4%, 62.5%, 

and 100% in patients with ACLF grade 1, ACLF grade 2 and 

ACLF grade 3, respectively, which was significant higher in 

comparison with patients in CANONIC study in corresponding 

grade (22.1% in grade 1, 32% in grade 2 and 76.7% 

in grade 3, respectively). The results indicated that the EASL- 

CLIF criteria may be too strict for diagnosis HBV-related ACLF. 

Of note, there were 21.4% of HBV-ACLF patients underlying 

chronic HBV hepatitis had more than one organ failure (assessing 

by CLIF-SOFA), and the mortality was 66.7% among them. 

The results suggested that diagnosis of HBV-ACLF should not 

exclude those patients without cirrhosis. Conclusions: ESAL-CLIF 

diagnostic criteria may not be suitable for assessing the patients 

with HBV-related ACLF in our study. A large, prospective, and 

multicenter study is needed to further validate the criteria. 

Disclosures: 

The following authors have nothing to disclose: Zhihong Wan, Chen Li, Shaoli 

You, HongLing Liu, ShaoJie Xin, Bing Zhu 

1610 

Anti-platelet therapy reduces the incidence of hepatocellular 

carcinoma in patients with chronic hepatitis B 

Minjong Lee 2 , Jeong-Hoon Lee 2 , Sohee Oh 1 , YOUNG CHANG 2 , 

Joon Yeul Nam 2 , Young Youn Cho 2 , Jeong-Ju Yoo 2 , Yuri Cho 2 , 

Donghyeon Lee 2 , Eun Ju Cho 2 , Su Jong Yu 2 , Yoon Jun Kim 2 , Jung- 

Hwan Yoon 2 ; 1 Department of Biostatistics, Seoul Metropolitan 

Government Seoul National University Boramae Medical Center, 

Seoul, Korea (the Republic of); 2 Department of Internal Medicine 

and Liver Research Institute, Seoul National University College of 


Background: Platelets contribute to liver damage by promoting 

the intrahepatic accumulation of virus-specific CD8 T cells in 

mouse models of viral hepatitis. A recent study showed that 

the long-term use of the anti-platelet drugs in a mouse model 

of chronic hepatitis B can prevent hepatocarcinogenesis. We 

aimed to compare the incidence of hepatocellular carcinoma 

in patients not treated with anti-platelet drugs to those treated 

with anti-platelet drugs. Methods: We performed a retrospective 

analysis of data from 3,479 consecutive patients with 

chronic hepatitis B who had HBV DNA completely suppressed 

with antiviral treatment. The population was divided into two 

groups: Group 1(n=2,891) not treated with anti-platelet drugs; 

Group 2 (n=588) treated with aspirin and/or clopidgrel. Data 

were collected from patients analyzed by a multivariable Cox 

proportional hazards model for the entire cohort. Results: 

During the study period, hepatocellular carcinoma was developed 

in 229 patients (7.9%) in Group 1 and 15 (2.5%) in 

group 2. In multivariable analyses, Group 2 patients showed 

a significantly lower risk of HCC than that in Group 1 (hazard 

ratio [HR]=0.21, 95% confidence interval[CI]=0.11–0.39, 

P5 ×ULN) were excluded 

due to potential overestimation of fibrosis by TE. Results: Serum 

M30 levels were significantly increased in significant fibrotic 

patients compared with no/minor fibrosis or NC (259.9 [IQR: 

122.6-501.2] vs 70.6 [IQR: 41.3 - 105.3] U/L, P


Disclosures: 

The following authors have nothing to disclose: ZhuJun Cao, Hui Wang, Xiaogang 

Xiang, FengDI Li, KeHui Liu, Weiliang Tang, Yuhan Liu, Lanyi Lin, Qing 

Guo, Qing Xie 

1612 

Statin and the risk of hepatocellular carcinoma and 

death in a hospital-based hepatitis B-infected population: 

a propensity-score landmark analysis 

John C Hsiang, Grace LH Wong, Vincent W. Wong, Henry Lik- 

Yuen Chan; Chinese University of Hong Kong, Hong Kong, Hong 

Kong 

BACKGROUND & AIMS The use of statin in hepatocellular carcinoma 

(HCC) and death prevention is still uncertain among 

hepatitis B-infected (HBV) patients. This study aimed to examine 

the effect of statin on HCC and death in a hospital-based HBV 

population METHODS We conducted a population study of 

HBV patients using the Hospital Authority registry database 

containing data of patients attending 43 public hospitals in 

Hong Kong. We defined statin use by landmark analysis 

to abrogate “immortal time bias” and propensity score (PS) 

weighting to minimise baseline confounders and “indication 

bias”. Multiple imputation analysis were performed for missing 

laboratory data. The weighted Cox regression analyses 

was performed for the risk of HCC (adjusting for competing 

mortality) and death. RESULTS A total of 73,499 patients, with 

a crude HCC incidence of 1.75 per 100 patient-years, were 

entered into the 2-year landmark analysis. After landmark analysis 

and PS weighting of baseline covariates, statin users had 

32% risk reduction in HCC (weighted sub-hazard ratio (SHR) 

0.68; 95% CI 0.48-0.97, p=0.033). In a PS weighted cohort 

of statin users and non-users, there was no difference in mortality 

compared statin users to non-users (weighted HR 0.92; 

0.76-1.11, p=0.386). In subgroup analysis, concurrent statin 

and nucleos(t)ide analogue (NA) use was associated with 

59% risk reduction in HCC (weighted SHR 0.41; 0.19-0.89, 

p=0.023) compared to NA use alone. CONCLUSION In this 

HBV cohort adjusted for confounders and biases, statin use 

is associated with reduced HCC risk by 32%. Additive HCC 

chemopreventive effect was seen with the concomitant use of 

NA and statin. Further prospective studies are warranted to 

investigate the potential use of statin in HBV-infected NA users. 

Weighted cumulative incidence of hepatocellular carcinoma 

(2-year landmark analysis, for a single multiple imputation dataset); 

p=0.033 

Disclosures: 

Grace LH Wong - Advisory Committees or Review Panels: Otsuka, Gilead; 

Speaking and Teaching: Echosens, Furui, Gilead, Janssen, Bristol-Myers Squibb, 

Otsuka, Abbvie 



Echosens 




The following authors have nothing to disclose: John C Hsiang 

1613 

Influence of the ethnic status in chronic hepatitis B 

patients: comparing the Netherlands, Belgium and Turkey 

Özgür M. Koc 1 , Sarah Konsten 1 , Geraldine Jansen 1 , Geert 

Robaeys 2,3 , Beytullah Yildirim 4 , Dirk Posthouwer 5 , Ger H. Koek 6 ; 

1 Faculty of Health, Medicine and Life Sciences, University Maastricht, 

Maastricht, Netherlands; 2 Department of Gastroenterology 

and Hepatology, East Limburg Hospital, Genk, Belgium; 3 Faculty 

of Medicine and Life Sciences, Limburg Clinical Research 

Program, Hasselt, Belgium; 4 Department of Gastroenterology, 

Ondokuz Mayis University, School of Medicine, Samsun, Turkey; 

5 Department of Internal Medicine, Infectious Diseases and Medical 

Microbiology, Maastricht University Medical Centre, Maastricht, 

Netherlands; 6 Department of Internal Medicine, Division of Gastroenterology 

and Hepatology, Maastricht University Medical Centre, 

Maastricht, Netherlands 

Background & Aims Hepatitis B virus (HBV) is a global threat 

affecting different ethnic groups. In some diseases, ethnicity 

is an essential predictor of disease outcome. Therefore, this 

study aimed to understand the influence of ethnic status on the 

natural history of chronic hepatitis B (CHB) infections in the 

Netherlands, Belgium and Turkey. Methods In this multicentre 

retrospective cohort study, 269 CHB patients from the Hepatology 

Outpatient Departments of three hospitals one in the 

Netherlands, Belgium and Turkey were included. Variables as 

baseline characteristics, route of transmission, laboratory characteristics 

and HBV endpoints were collected. Ethnicity was 

defined as country of birth. Chi-square and ANOVA tests were 

used for categorical and continuous variables, respectively. 

Results The CHB population in the Netherlands, Belgium and 

Turkey consisted of respectively 98, 68 and 103 patients. In 

the Netherlands 32.7 and 16.3% were respectively of Dutch 

and Chinese origin. In Belgium mainly Belgian (36.8%) and 

Turkish (33.8%) patients were included, whereas the population 

in Turkey consisted in 99% of patients of Turkish origin. 

The CHB population in the Netherlands, Belgium and Turkey 

differed significantly in the number of vertical (40.8, 17.6 and 

15.5% respectively, p= .000), sexual (16.3, 13.2 and 3.9% 

respectively, p= .013) and intra-familial transmission (1.0, 

11.8 and 26.2% respectively, p= .000). In addition, the countries 

showed significant differences in HBV endpoints such as 

the number of cirrhosis (11.2, 17.6 and 38.8% respectively, 

p= .000) and hepatocellular carcinoma (2.0, 2.9 and 11.7% 

respectively, p= .008). Subsequently, patients from Belgium 

and the Netherlands were categorized as of Dutch / Belgian 

(n=46), Turkish (n=27) and Chinese (n=18) origin. There was 

a significant difference in the number of vertical transmission 

(12.1, 22.2 and 83.3% respectively, p= .000), sexual transmission 

(27.6, 7.4 and 0.0% respectively, p= .007) and 

cirrhosis (8.6, 22.2 and 0.0% respectively, p= .045). Conclusions 

The Netherlands, Belgium and Turkey differed regarding 

ethnicity, routes of transmission and disease progression. Chinese 

patients were characterized by vertical transmission and


absence of cirrhosis. In addition, Turkish and Dutch/Belgian 

patients had significantly more cirrhosis and acquired the disease 

by intra-familial and sexual transmission, respectively.This 

study therefore implies the importance of ethnicity as there is an 

association between ethnic groups, routes of transmission and 

disease progression. 

Disclosures: 

Geert Robaeys - Advisory Committees or Review Panels: Gilead; Speaking and 

Teaching: Merck, Janssens 

The following authors have nothing to disclose: Özgür M. Koc, Sarah Konsten, 

Geraldine Jansen, Beytullah Yildirim, Dirk Posthouwer, Ger H. Koek 

1614 

High-throughput and ultrasensitive assay for the quantification 

of HBV precore and basal core promoter 

mutants and their clinical significance 

Motokazu Mukaide, Kazumoto Murata, Masaya Sugiyama, 

Masashi Mizokami; National Center for Global Health and Medicine, 

Ichikawa, Japan 

Background/Aim: Precore (PC) G1896A and basal core promoter 

(BCP) A1762T/G1764A mutations of HBV is associated 

with HBeAg seroconversion. However, in the previous 

studies, the percentage of PC and BCP mutant were examined 

by semi-quantitative assay, and lacked sufficient sensitivity, 

which make difficult for precise prediction for the response to 

therapies in each patient. Therefore, our aim of this study is 

to develop a novel quantitative assay for HBV, with special 

reference to PC and BCP, using new generation nanoliter-sized 

droplet technology paired with digital PCR (ddPCR), and to 

investigate its clinical significances. Methods: We collected 

serum samples from 95 HBV patients [mean: 54 years, sex 

male 69%, ALT 45U/L(range 14 to 333), DNA 3.6 logIU / 

mL, HBeAg positive 27.4%, HBV genotypes (B: 11, C: 84) . 

Serum samples were stored at -80°C until use. Among them, 

57 patients were under treatment with nucleos(t)ide analogs 

and no patients were treated with IFN. ddPCR assays were 

performed using the QX100 ddPCRsystem. ddPCR primers and 

probes were designed on the basis of the conserved nature of 

these sequences using the Hepatitis Virus Database. Results: 

The assay using standard plasmid linearly detected HBV from 

2.5 to 10 5 copies, and the limit of detection was 0.005% of 

mutant (MT) in the presence of wild-type (WT). The total viral 

load (MT+WT) quantified by ddPCR showed a linear correlation 

with HBV DNA levels as measured by COBAS TaqMan 

HBV Test (range 2.1 to 9.1 log 10 

IU/mL: PC: R 2 =0:86, P


factors independently predict immune reactive phase, i.e. alanine 

aminotransferase (< 80 U/L, 0; ≥ 80 U/L, 1), HBV DNA 

(≥ 8 log IU/mL, 0; 6-8 log IU/mL, 2; < 6 log IU/mL, 3) and 

HBsAg (> 4.25 log IU/mL, 0; < 4.25 log IU/mL, 3); and a prediction 

score ranging from 0 to 7 was established. Cutoff values 

of 4 best discriminated immune reactive phase with area 

under the receiver operating characteristic curves (AUROC) of 

0.830 (0.794-0.862) in training cohort. By applying the cutoff 

value of 4, the score differentiates immune reactive phase with 

AUROC of 0.812 (0.757-0.859) and positive predictive value 

of 89% in validation cohort, respectively. Conclusion: A simply 

non-invasive diagnostic score constructed from routine laboratory 

parameters is accurate in differentiating immune phases 

in HBeAg-seropositive persons. Future validation is warranted. 

Disclosures: 

The following authors have nothing to disclose: Qing-Lei Zeng, Yan-Ling Fu, Jun 

Lv, Zu-Jiang Yu 

1617 

Postpartum hepatic flare after telbivudine withdrawal in 

pregnant women with chronic hepatitis B 

Jinfeng Liu 2 , Jing Wang 1 , Dongfang Jin 3 , Yingli He 2 , Li Jin 1 , Taotao 

Yan 1 , Furong Cao 1 , Yuan Yang 2 , Shulin Zhang 2 , Yingren Zhao 2 , 

Tianyan Chen 2 ; 1 Xi’an Jiaotong University, Xi’an, China; 2 The First 

Affiliated Hospital of Medical College, Xi’an Jiaotong University, 

Xi’an, Shaanxi Province, China, Xi’an, China; 3 Shaanxi Kangfu 

Hospital, Xi’an, China 

Background: The efficacy of telbivudine for breaking vertical 

transmission of HBV has been established. Data on the risk of 

postpartum flare after telbivudine withdrawal and optimal duration 

of antiviral therapy after delivery are limited. Methods: 

We enrolled 312 women who received telbivudine beginning 

at week 24 or 28 of gestation and then followed up to 52 

weeks postpartum. Virological and biochemical parameters 

were assessed. Results: Of the 312 women enrolled, 26.3% 

had elevated serum alanine aminotransferase (ALT) during 

pregnancy. Telbivudine administration resulted in ALT normalization 

in 80.5% before delivery. Compared with women 

having a normal ALT level throughout pregnancy, those with 

elevated ALT had a significantly higher rate of ALT flare after 

telbivudine withdrawal (27.8% vs. 7.0%; P = 0.032). Multivariate 

analysis indicated only ALT elevation during pregnancy 

correlated with postpartum flare after telbivudine withdrawal. 

Those women with elevated ALT during pregnancy, continued 

antiviral treatment to 52 weeks postpartum, had a significantly 

higher HBeAg seroconversion rate (P = 0.008) and a notable 

decrease in HBsAg titers (P = 0.046). Conclusion: It is safe for 

the majority of women to withdraw telbivudine after delivery; 

whereas its withdrawal is not prudent for women with elevated 

ALT during pregnancy. 

Disclosures: 

The following authors have nothing to disclose: Jinfeng Liu, Jing Wang, Dongfang 

Jin, Yingli He, Li Jin, Taotao Yan, Furong Cao, Yuan Yang, Shulin Zhang, Yingren 

Zhao, Tianyan Chen 

1618 

Performance Assessment of Common Anti-hepatitis B 

Core Antigen Assays in Japan for Prevention of HBV 

Reactivation 

Takako Inoue 1 , Tomoya Iwase 1 , Satomi Kani 1 , Kumiko Ohne 1 , 

Takaaki Goto 1 , Yukio Wakimoto 1 , Yasuhito Tanaka 2,1 ; 1 Clinical 

Laboratory, Nagoya City University Hospital, Nagoya, Japan; 

2 Department of Virology & Liver unit, Nagoya City University 

Graduate School of Medical Sciences, Nagoya, Japan 

Background: The measurement of anti-hepatitis B core antigen 

(anti-HBc) is a significant method to predict the risk of hepatitis 

B virus (HBV) reactivation in patients with immunosuppressive 

drug therapies. In Japan, several commercially assays for anti- 

HBc are available. The aim of this study is to compare the 

anti-HBc measurement methods and to evaluate those clinical 

utilities. Methods: Three kinds of anti-HBc assays were evaluated 

with 77 clinical samples which showed low anti-HBc titers 

around the cutoff values (from 0.1 to 2.0 C.O.I). The anti-HBc 

assays used in this study were two chemiluminescent enzyme 

immunoassay (HISCL anti-HBc [Sysmex] and Lumipulse anti- 

HBc N [Fujirebio, Inc.]) and one chemiluminescent immunoassay 

(Architect HBc II [Abbott Japan]). This study protocol was 

approved by the appropriate institutional ethics review committees. 

Results: In the direct comparison between HISCL anti- 

HBc and Architect HBc II, the concordance rate was 79.2% 

(61/77). Between HISCL anti-HBc and Lumipulse anti-HBc N, 

the concordance rate was 80.5% (62/77). Between Lumipulse 

anti-HBc N and Architect HBc II, the concordance rate was 

96.1% (74/77). In the 77 clinical samples, there were 17 

discrepant samples (TABLE). Of 9 discrepant samples negative 

only by HISCLE, 6 samples were positive for anti-HBs without 

the history of HB vaccination, indicating that they have been 

exposed to HBV. Of 8 discrepant samples positive by HISCLE, 

7 samples were positive by absorption test for HISCLE anti- 

HBc, implying that the results by Lumipulse and Architect were 

false negative. One of the discrepant samples positive only for 

HISCLE anti-HBc was HBsAg seropositive, and HBV-DNA was 

also detected (chronic hepatitis B carrier). Conclusions: In this 

study, the results of anti-HBc show discrepancies between each 

of the anti-HBc assays. In cases with near the cutoff values of 

anti-HBc, it would need to be retested with other assays or 

molecular methods. Future studies should standardize the anti- 

HBc titer measurement system to prevent HBV reactivation. 

Discrepancies (17 samples) 

Disclosures: 



Squibb 

The following authors have nothing to disclose: Takako Inoue, Tomoya Iwase, 

Satomi Kani, Kumiko Ohne, Takaaki Goto, Yukio Wakimoto 

1619 

Does Cesarean section and formula feeding reduce 

vertical transmission in chronic hepatitis B pregnant 

mothers? 

Rasham Mittal, Amandeep K. Sahota; Kaiser Permanente Los 

Angeles Medical Center, Los Angeles, CA 

Background Perinatal acquisition of hepatitis B virus (HBV) carries 

85% to 95% risk of chronic infection. While few studies 

have supported cesarean section (C-section) and bottle-feed-


ing, others have challenged it. Limited data is available from 

United States. Study Aim: Effect of C-section and formula feeding 

on vertical transmission in a cohort of CHB pregnant mothers 

from United States. Methods: Retrospective study (January 

2005- June 2010) at a large integrated health care system in 

Southern California. CHB pregnant mothers identified using 

regional perinatal database. Inclusion criteria: age > 18 years, 

positive serum HBsAg (hepatitis B surface antigen). Exclusion 

criteria: negative/ false positive HBsAg; termination of pregnancy 

or lost to follow up. Data Collection; demographics, 

first live-birth pregnancy reviewed, mode of delivery, breast 

feeding (defined as breastfeeding for ≥2 weeks after birth), 

maternal hepatitis B e antigen (HBeAg), and HBsAg testing of 

infants at 9-18 months of age. Lost to follow up if child not seen 

at 9-18 months of age. Fisher’s exact test used to determine 

statistical significance. Results: A total of 472 infants were born 

to 462 CHB mothers (mean maternal age 32.5 years, range 

18-45; 78.8% Asian-Pacific Islanders). 308 (66.7%) delivered 

vaginally and 154 (33.3%) through C-section. All C-sections 

were performed for obstetrical indications, with labor arrest 

being most common. 384 (83.1%) mothers breast-fed after 

delivery. Mode of feeding could not be established for 13 

(2.8%) mothers. Hepatitis Be antigen (HBeAg) status of mothers 

outlined in table 1. Only 1 infant (delivered vaginally and 

breast-fed) born to HBeAg positive mother (unknown antenatal 

HBV viral load) had reactive HBsAg. No statistically significant 

difference found for risk of vertical transmission between 

mothers who breastfed vs. formula feeding (1/230 vs. 0/35; 

p>.05); normal vaginal vs. cesarean-section delivery (1/185 

vs. 0/85; p>.05). Limitation of data: 104 (26.2%) infants did 

not have HBsAg tested at 9-18 months follow up. Missing antenatal 

HBeAg and HBV viral load data for mothers. Conclusion: 

C-section and formula feeding did not reduce vertical transmission 

risk in cohort of CHB pregnant mothers from United States. 

Table 1 Maternal practices: Infant feeding, delivery mode 

HBV-related chronic liver diseases needs to be further studied, 

especially regarding the role of hepcidin. Methods: In the 

present study, we investigated the association between serum 

hepcidin, iron metabolism and the clinical indictors in the 

patients with HBV-related chronic liver diseases in a case-control 

study(2013-2015). A total of 318 subjects were included 

in the study, including 78 cases with HBV-related chronic hepatitis, 

85 cases with HBV-related liver cirrhosis, 77 cases with 

HBV-related hepatocellular carcinomar(HCC), and 78 healthy 

individuals as controls. Results: Compared with the healthy controls, 

all groups of cases had significantly higher levels of serum 

ferritin, and much lower levels of serum hepcidin. Compared 

with HBV-related chronic hepatitis, the levels of HBV-DNA loading, 

serum iron, total iron binding force(TIBC) and serum transferrin 

were more lower in the patients with cirrhosis and HCC, 

and the serum hepcidin levels were obviously higher. The level 

of serum ferritin was increased with the severity of liver fibrosis 

and inflammation activity. The multiple linear regression analysis 

showed that serum hepcidin levels had obvious correlation 

with HBV-DNA loading levels (β=-0.277, t=-4.438, P


.Only results obtained with at least ten successful acquisitions, 

success rate of at least 60% and IQR/LSM ratio less than 30% 

were considered reliable. Metavir liver fibrosis stages were 

assessed by the same pathologist experienced blinded to the 

results of LSM. Statistical analysis was made according to SAS. 

RESULTS: Statistical analysis was conducted in 109 patients, 

26 patients were excluded for unreliable LB or LSM. Sex ratio 

M/F was 1.65, mean age was 38.6 years. The mean biopsy 

length was 26.4mm. The distribution of liver fibrosis according 

to the METAVIR score were: F0 (n = 17) (15.5%),F1: n = 51 

(46.7% ) F2: n = 20 (18.3%), F3: n = 15 (13.7%) and F4: n 

= 6 (5.5%). The mean delay between LSM and LB was 6 days, 

the mean acquisition success rate was 95% and the median 

IQR / LSM ratio was 14.6%. The area under receiver –operating 

characteristic curves were 0.87 (95%confidence intervals 

0.80-0.94) for F≥2, 0.90 (0.82-0.98) for F≥ 3 and 0.93 

(0.83-1)for F4.Optimal LSM cut-off values were 7.1 for ≥F2,8.6 

for≥F3 and 12 for F4 by maximizing of sensitivity and specificity.In 

univariate analysis, LSM was significantly correlated with 

Metavir fibrosis stage,activity grade,gender, and biopsy length 

.In multivariate analysis, LSM was significantly correlated only 

with Metavir fibrosis stage,activity grade, and biopsy length. 

CONCLUSION: LSM is an accurate method for assessment liver 

fibrosis in patients with chronic hepatitis B e negative with high 

performance diagnostic for cirrhosis. 

Disclosures: 

Salah Sahraoui - Employment: bms, roche 

Nabil Debzi - Speaking and Teaching: roche, msd, bms, Janssen, Abbvie 

The following authors have nothing to disclose: Nawal Guessab, Nawel Afredj, 

Rafik Kerbouche, Ibtissem Ouled Cheikh, Sonia Ait Younes, Samir Gourari, 

Yazid Chikhi, Salima Cheraitia, Mohamed Merniz, François Montestruc, Saadi 

Berkane 

1622 

Evaluation of HBV infection status in spouses of 

HBsAg-positive individuals 

Ebru Dik, Selma Tosun, Seher A. Coskuner, Alpay Ari, Meltem 

Avci; Infectious Diseases, Bozyaka Education and Research Hospital, 

Izmir, Turkey 

Objective: The transmission of infection through sexual intercourse 

is one of the most common routes of transmission for 

hepatitis B virus. Therefore the spouses of HBsAg-positive 

individuals must be screened for HBsAg positivity. The aim 

of the present study was to evaluate HBV infection status in 

spouses of HBsAg-positive individuals. Method: Using face-toface 

interview technique, a questionnaire was administered to 

HBsAg-positive cases who were followed in Viral Hepatitis Outpatient 

Clinics at zmir Bozyaka Education and Research Hospital 

in order to evaluate HBV infection status in their spouses, 

and the results were recorded if tests were available. Results: 

The study included a total of 949 HBsAg-positive cases who 

were aged 17 to 76 years. The spouses of 409 (43%) cases 

were never tested for HBV infection. When the results of the 

tested individuals were evaluated, the rate of HBsAg positivity 

was 6% (n=32), rate of antiHBcIgG+AntiHBs positivity was 

18%(n:96), rate of antiHBs positivity was 12%, rate of antiHBcIgG 

positivity alone was 1%, and rate of negative results in all 

HBV parameters was 62%. Conclusion: The data suggest that 

not all spouses of the individuals were tested for HBV infection. 

It is a striking finding that HBV infection status was unknown 

in 43% of the spouses. The rate of HBsAg positivity was 6% 

and the rate of immunization was very low (12%). AntiHBcIg- 

G+AntiHBs positivity was 18%. These results suggest that HBV 

transmission to the spouses from HBsAg-positive individuals is 

common, but the awareness of the individuals was very low. 

Low rate of testing and high rate of HBsAg positivity among 

the spouses suggest that screening of spouses for HBV infection 

must not be neglected. 

Disclosures: 

The following authors have nothing to disclose: Ebru Dik, Selma Tosun, Seher A. 

Coskuner, Alpay Ari, Meltem Avci 

1623 

Is booster beneficial after universal HBV vaccination? 

Selma Tosun 1 , Erhun Kasirga 2 ; 1 Infectious Diseases, Bozyaka Education 

and Research Hospital, Izmir, Turkey; 2 Pediatric Gastroenterology, 

Celal Bayar University The Faculty of Medicine, Manisa, 

Turkey 

Objective: The infants are administered hepatitis B vaccination 

after birth at 0, 1, and 6 months. HBV vaccine-induced 

antibody titers are maintained in the blood for a long time 

(possibly lifelong) in immunocompetent individuals. However, 

there are some studies suggesting a considerable decrease 

in antiHBs titers below protective level 7 to 8 years after HBV 

vaccine administered immediately after birth. The studies also 

indicate an excellent antiHBs response with a booster dose 

of HBV vaccine. The aim of the present study was to evaluate 

antiHBs response in children with or without a booster vaccine 

dose after universal HBV vaccination. Method: The study 

included primary school children who received HBV vaccine 

after birth. The study subjects were divided into 2 groups and 

of the students in Group 1, 157 received three booster doses of 

HBV vaccine 7 to 9 years after national vaccination program. 

Totally 142 students in Group 2 did not receive any vaccine 

after the initial HBV vaccines administered after birth. AntiHBs 

titers were studied in blood samples, and the students with a 

antiHBs titer below 10 IU/mL were also tested for HBsAg and 

antiHBcIgG . Results: The study included a total of 299 student 

aged 7 to 12 years. The rate of positive antiHBs titers in students 

that received three booster doses of vaccine in Group 1 

was statistically significantly higher compared to the students 

in the second group (p


1624 

Assessment Of Treatment Response By Elastography 

During The Tenofovir Treatment In Nucleos(t)ide-Naïve 

Chronic Hepatitis B Patients 

Taeheon Lee, Byung Ik Kim, Yong Kyun Cho, Woo Kyu Jeon, Hong 

Joo Kim; Kangbuk Samsung Hospital, Seoul, Korea (the Republic 

of) 

Aim: Several longitudinal studies have demonstrated that elastrograpy 

could be a useful tool for monitoring of liver fibrosis 

during the treatment of chronic hepatitis B. However, not 

enough studies assessing regression of liver fibrosis by elastography 

are available to date. Therefore, the purpose of this study 

was to evaluate the liver stiffness before and during the treatment 

and assess the usefulness of this non-invasive modality. 

Methods: Fourty-eight treatment-naïve patients started treatment 

with tenofovir. Liver stiffness measurement by a shear wave 

elastography and a conventional ultrasonography was performed 

at baseline, after median 6 and 12 months. Cut-off levels 

to Metavir classification for fibrosis stage F0-1, F2, F3 and 

F4 were ≤5.6 kPa, ≤7.44 kPa, ≤8.71 kPa, and ≥10.87 kPa, 

respectively. Result: Median liver stiffness decreased from 6.68 

kPa(SD, ±3.90), 6.27(SD, ±3.88) and 5.69 kPa(SD, ±5.01) in 

patients treated with tenofovir at baseline, after median 6 and 

12 months. There were no significant difference in the degree 

of regression of liver stiffness after median 6 (P =0.29) and 12 

months (P =0.14). Twenty-two(45.9%) subjects were classified 

as F0-1, 3 (6.3%) as F2, 20 (41.7%) as F3, and 3 (6.3%) as 

F4. After median 6 and 12 months later, liver stiffness value of 

patients recorded as F0-1 was decreased mean 0.60 kPa(SD, 

±2.38) and 0.05 kPa(SD, ±2.17), F2 as 0.81 kPa(SD, ±1.69) 

and 0.99 kPa(SD, ±3.01), F3 as 0.81 kPa(SD, ±3.75) and 

1.02 kPa(SD, ±5.67)a and F4 as 13.23 kPa(SD, ±7.59) and 

4.50 kPa (P for trend =0.01 and 0.04 after 6months and 

12 months) Conclusion: Liver stiffness was not significantly 

improved during the tenofovir treatment in patients with chronic 

hepatitis B during 6 and 12 months. But these result showed a 

trend that in a patient with advanced fibrosis on baseline stiffness 

stage was more improvedcompared to that of mild fibrosis 

during 6 and 12 months. Long term prospective studies are 

required to investigate the usefulness of elastography for relationship 

between elastography stiffness and treatment response 

in chronic hepatitis B patients treated with tenofovir. 

Disclosures: 

The following authors have nothing to disclose: Taeheon Lee, Byung Ik Kim, Yong 

Kyun Cho, Woo Kyu Jeon, Hong Joo Kim 

1625 

Inadequate Treatment of Chronic Hepatitis B in Pregnant 

Women at High Risk for Vertical Transmission 

Diana Y. Wu 1,2 , muhammad I. Shafi 3,2 , Mohammed M. Aboelsoud 

3,2 , Colin Woodard 4,2 ; 1 Gastroenterology, Women and 

Infants Hospital, Providence, RI; 2 Brown University, Alpert Medical 

School, Providence, RI; 3 Internal Medicine, Memorial Hospital, 

Pawtucket, RI; 4 Internal Medicine, Kent Hospital, Warwick, RI 

Aim: To describe the clinical presentation and management of 

a cohort of pregnant women with hepatitis B in a single center 

specializing in obstetrics care. Methods: Pregnant women 

with positive hepatitis B surface antigen who were seen at 

Women and Infants Hospital, Providence, Rhode Island from 

2009-2014 were included. Medical records were reviewed for 

demographic and clinical information from initial presentation 

for positive hepatitis B surface antigen until time of delivery. 

Results: 112 women were included, median age 28 yrs. Of the 

76 women who had their birth countries recorded in their medical 

record, only seven (9%) were born in the United States. 

Median follow up was 4.8 months. 19 (17%) women had ALT 

levels more than twice the upper limit of normal for women (19 

U/L) during their pregnancy. 20 (18%) women were hepatitis 

B e-antigen positive. 15 women (13%) had HBV DNA levels 

> 200,000 IU/ml in the third trimester of pregnancy, which 

qualified them for initiation of treatment for chronic hepatitis B 

to prevent vertical transmission to their fetus. Five of these pregnant 

women were initiated on Lamivudine, three on Tenofovir, 

and one on Adefovir in their third trimesters. However, five of 

the eligible 15 (33%) women were not initiated on treatment 

during pregnancy to prevent vertical transmission, and a sixth 

woman declined therapy. Two of the 112 women in the study 

were already on therapy for chronic hepatitis B before pregnancy, 

one on Lamivudine and one on Adefovir, which were 

continued throughout pregnancy. One woman was on Entecavir 

before pregnancy and was not switched to a different oral 

agent during pregnancy. Another woman on Entecavir before 

pregnancy was switched to Telbivudine during pregnancy. 

Conclusions: Although there is universal screening of pregnant 

women in prenatal care in the US for chronic hepatitis B, there 

remains inadequate initiation of treatment in the third trimester 

for patients with high viral load, who are at high risk of vertical 

transmission. Furthermore, this study shows that there should 

be improved knowledge of safe oral antiviral therapies for 

hepatitis B in pregnancy. 

Disclosures: 

The following authors have nothing to disclose: Diana Y. Wu, muhammad I. 

Shafi, Mohammed M. Aboelsoud, Colin Woodard 

1626 

NK cell function is restored with sequential NUC therapy 

and correlates with treatment response in Chronic Hepatitis 

B 

Upkar S. Gill 1 , Dimitra Peppa 2 , Lorenzo Micco 2 , Harsimran D. 

Singh 2 , Ivana Carey 3 , Graham R. Foster 1 , Mala K. Maini 2 , Patrick 

T. Kennedy 1 ; 1 Hepatology Unit, Centre for Immunobiology, 

Blizard Institute, Barts and The London, School of Medicine & Dentistry, 

QMUL, London, United Kingdom; 2 Division of Infection & 

Immunity, UCL, London, United Kingdom; 3 Institute of Liver Studies, 

Kings College London, London, United Kingdom 

INTRODUCTION: New treatment strategies to achieve HBsAg 

loss in Chronic Hepatitis B (CHB) are required. Sequential/ 

combined therapeutic approaches comprising Pegylated-Interferon 

(Peg-IFNα) and nucleot(s)ide analogues (NUC) are being 

given greater consideration. We previously demonstrated 

boosting of NK cells in eAg- patients treated with Peg-IFNα 

(Micco et al, J. Hepatol, 2013). Here we studied differential 

NK cell responses in patients receiving sequential NUC therapy, 

after Peg-IFNα exposure, to elucidate a possible treatment 

advantage over current therapy strategies. PATIENTS & 

METHODS: PBMC from 43 eAg+ patients were analysed. 21 

patients underwent a 48-week course of Peg-IFNα and were 

studied longitudinally; 12/21 progressed to sequential NUC 

therapy and were followed until viral suppression. 12 patients 

receiving de novo NUC therapy, without Peg-IFNα exposure 

were compared for analysis. In addition 10 patients exposed 

to Peg-IFNα, and not progressing to sequential NUC therapy, 

were also studied for comparison. Phenotypic and functional 

analysis of NK cell subsets was performed by multicolour 

flow-cytometry and findings correlated with on-therapy HBsAg 

changes. RESULTS: There was cumulative expansion of activated 

and functional CD56 bright NK cells throughout 48-weeks 

of Peg-IFNα (p=0.0001), maintained at higher than pre-treatment 

levels for at least 9-months after switching to sequential


NUCs. Proliferating NK cells with antiviral capacity increased 

significantly, and HBsAg significantly decreased, when NUC 

treatment was preceded by Peg-IFNα compared to de-novo 

NUCs. Interestingly, the frequency of antiviral NK cells, in 

those patients exposed to Peg-IFNα, without further therapy, 

reduced to baseline, 9-12 months post treatment cessation. 

In addition, increased IFNγ production and cytotoxicity, but 

reduced TRAIL expression, were noted in those patients (9/12) 

with HBsAg decline on sequential NUCs. CONSLUSIONS: 

The potent and cumulative expansion of activated NK cells 

with antiviral potential induced by Peg-IFNα is maintained on 

sequential NUCs. Patients receiving sequential NUCs achieved 

a greater decline in HBsAg than those treated with de novo 

NUCs, particularly those with TRAIL low IFNγ/CD107 hi NK cells. 

TRAIL expression in this setting may be pathogenic, in line with 

our previous data showing that TRAIL+ NK cells can delete 

antiviral T cells (Peppa et al, JEM 2013), suggesting the TRAIL 

pathway as a potential future therapeutic target. These findings 

provide mechanistic insights to support further consideration of 

sequential Peg-IFNα/NUC therapy to enhance HBsAg reduction, 

and achieve sustained treatment responses. 

Disclosures: 


BMS 







Patrick T. Kennedy - Grant/Research Support: Roche, Gilead; Speaking and 

Teaching: BMS, Roche, Gilead 

The following authors have nothing to disclose: Upkar S. Gill, Dimitra Peppa, 

Lorenzo Micco, Harsimran D. Singh, Mala K. Maini 

1627 

Analysis of the effect on HBV life cycle by HBV genome 

editing using two different CRISPR/Cas9 systems 

Hiromi Abe, Keiichi Masaki, Tetsushi Sakuma, Masataka Tsuge, 

Nobuhiko Hiraga, Michio Imamura, C. Nelson Hayes, Hiroshi 

Aikata, Takashi Yamamoto, Kazuaki Chayama; Hiroshima University, 

Hiroshima, Japan 

Background and aim: Interferon and nucleoside analogue therapy 

effectively controls HBV replication in chronically infected 

patients. However, it is still difficult to eradicate HBV completely 

because covalently closed circular DNA (cccDNA) stably 

remains in the nucleus of hepatocytes. Recently, a new 

genome editing system, Cas9-nickase, which has mutagenic 

Cas9 activity, has been developed to target specific regions 

of double stranded DNA sequences with less off target effects 

compared with the original CRISPR/cas9. Using this approach 

we developed an anti-HBV system that targets three different 

portion of the HBV genome using only one plasmid. The aim 

of this study is to investigate the efficacy of these two CRISPR/ 

Cas9 systems. Method: Two CRISPR/cas9 plasmids, nuclease 

and nickase types, were constructed to target three regions 

of the HBV genome: HBs, polymerase, and core. The original 

CRISPR-nuclease digests the HBV genome at these three 

regions using three target guide RNAs whereas the nickase 

type enzyme nicked both strands of the target site DNA using 

three pairs of guide RNAs transcribed from one plasmid. 

HepG2 cells grown in 6 well-plates in DMEM containing 10% 

FBS were co-transfected with 1.4×HBV genome and one of 

the two CRISPR-Cas9 encoding plasmids in a 1:2 ratio. Three 

days after co-transfection, cells and culture medium were harvested 

to evaluate the efficacy of the genome editing. Core 

associated HBV DNA was measured by qPCR after immunoprecipitation 

with anti-HBc antibody. The CRISPR/Cas9 plasmid 

was introduced into Baculovirus infection system. 14 days after 

infection, we measured HBsAg, eAg, and HBV DNA in the 

supernatant and intracellular HBV DNA and core-associated 

HBV DNA. Results: More than 78-95% of double stranded 

DNA of HBs, core and polymerase cleavage by both nuclease 

and nickase activity of CRISPR/Cas9 were confirmed by reporter-based 

assay. When we co-transfected 1.4×HBV genome 

and each CRISPR/Cas9-encoding plasmid, both HBsAg and 

eAg levels declined significantly in comparison with control 

plasmids in both types of CRISPR/Cas9 systems. Nuclease type 

Cas9 more efficiently reduced both HBsAg and eAg levels than 

nuclease type to less than 10% of control. Cas9-nickase also 

reduced HBsAg and eAg to approximately 10% of control cells 

(P


HBV core-related antigen level. In the human hepatocytes, 

HDI decreased HBs antigen level by 70%, HBV DNA level 

by 90% and 3.5kb HBV RNA level by 50% in the cell. HDI 

increased HBs antigen level in the HepG2 cells transfected 

with the entire HBV genome and also increased 3.5 kb HBV 

RNA level by 3~4-folds. HDI increased HBV DNA associated 

with acetylated histone H4 as well as acetylation of histone H4 

in the HepG2.2.15 cells. Furthermore, histone acetyltransferase 

inhibitor garcinol partially inhibited the increase in 3.5kb 

HBV RNA level by HDI indicating the possible role of histone 

acetylation on the regulation of HBV replication. Conclusion: 

HDI increases HBV replication but decreases HBs antigen production 

in hepatoma cell line with HBV DNA integration. In 

non-tumor hepatocytes infected with HBV, HDI inhibits HBV 

production as well as HBV replication. Epigenetic mechanism 

may be involved in the regulation of HBV replication at multisteps 

in human hepatocytes. 

Disclosures: 

Tatsuo Kanda - Grant/Research Support: MSD K.K.; Speaking and Teaching: 

AJINOMOTO PHARMACEUTICALS CO., LTD, CHUGAI PHARMACEUTICAL 

CO., LTD, GlaxoSmithKlein, BMS, Jansen, Daiichisankyo; Stock Shareholder: 

Mitsubishi Tanabe Pharma 

Osamu Yokosuka - Grant/Research Support: Chugai, Taiho, Bristol Myers, 

Takeda 

The following authors have nothing to disclose: Shingo Nakamoto, Shuang Wu, 

Yuki Haga, Reina Sasaki, Masato Nakamura, Hiroshi Shirasawa 

1629 

CRISPR/Cas9 “double”-nickase mediated inactivation of 

hepatitis B virus replication 

Madina Karimova 2 , Niklas Beschorner 2 , Werner Dammermann 1 , 

Jan Chemnitz 2 , Daniela S. Indenbirken 2 , Adam Grundhoff 2,3 , Stefan 

Lueth 1 , Frank Buchholz 4 , Schulze zur Wiesch Julian 1,3 , Joachim 

Hauber 2,3 ; 1 1. Department of Medicine, University Medical Center 

Hamburg-Eppendorf, Hamburg, Germany; 2 Heinrich Pette Institute 

– Leibniz Institute for Experimental Virology, Hamburg, Germany; 

3 German Center for Infection Research (DZIF), partner site Hamburg, 

Hamburg, Germany; 4 Department of Medical Systems Biology, 

University Hospital and Medical Faculty Carl Gustav Carus, 

TU Dresden, Dresden, Germany 

Background: Current antiviral therapies cannot cure hepatitis B 

virus (HBV) infection, since successful HBV eradication would 

require the inactivation of the viral genome, which primarily 

persists in host cells as episomal covalently closed circular 

DNA (cccDNA) and, to a lesser extent, as chromosomally integrated 

sequences. However, novel designer-enzymes, such as 

the CRISPR/Cas9 RNA-guided nuclease system, provide the 

technology for the development of advanced therapy strategies 

that directly attack the HBV genome. Methods: We report here 

the identification of cross-genotype conserved HBV sequences 

in the HbS and HbX region of the HBV genome that are specifically 

and effectively inactivated by a Cas9 double-nickase 

approach. Pairs of appropriately spaced Cas9 nickase 

mutants introduced two single-strand breaks on the opposite 

DNA strands that were subject to NHEJ repair in order to avoid 

off-target mutations by improving specificity by up to 1,500-fold 

relative to the wild-type Cas9 enzyme. Results: We show that 

this approach equally inactivated episomal cccDNA as well as 

chromosomally integrated HBV target sites in reporter cell lines 

as well as in chronically infected hepatoma cell lines. Analysis 

of Cas9n activity on ORF S and X target sites by next generation 

sequencing revealed efficient editing of cccDNA molecules 

targeted by either gRNA in HBV-infected HepG2.2.15, 

HepG2-H1.3 and HepG2-NTCP cells. The efficiency of X-specific 

sgRNAs was particularly high, with approximately 90% 

of all amplicons reads showing clearly discernible indel signatures. 

Conclusion: Our data support the feasibility of using 

the CRISPR/Cas9 nickase system for novel therapy strategies 

aiming to provide a cure for HBV infection. 

(A) Depiction of the HBV genome with location of amplicons 

spanning the S- and X-specific sgRNA target regions. The two 

amplicons are shown as dark gray boxes labeled S- and X-amplicon, 

respectively. Arrows shown at the top indicate the genomic 

location of gRNA target sequences. 

Disclosures: 

Stefan Lueth - Advisory Committees or Review Panels: BMS, Gilead, MSD, Janssen 

The following authors have nothing to disclose: Madina Karimova, Niklas Beschorner, 

Werner Dammermann, Jan Chemnitz, Daniela S. Indenbirken, Adam 

Grundhoff, Frank Buchholz, Schulze zur Wiesch Julian, Joachim Hauber 

1630 

GORASP2 regulates HCV and HBV through RNA and/or 

DNA replication and virion trafficking 

Dachuan Cai 1,2 , Jian Hong 1 , Xiao Liu 1 , Sae Hwan Lee 1 , Dahlene 

Fusco 1 , Esperance A. Schaefer 1 , Cynthia Brisac 1 , Anna Lidofsky 1 , 

Wenyu Lin 1 , Raymond T. Chung 1 ; 1 Gastrointestinal Unit, Massachusetts 

General Hospital, Harvard Medical School, Boston, MA; 

2 Department for infectious diseases, The second affiliated hospital 

of Chongqing medical university, Chongqing, China 

Background/Aims: Hepatitis B virus (HBV) and HCV constitute 

major causes of blood transmitted hepatitis. Chronic HCV 

and HBV infection induce liver cirrhosis and hepatocellular 

carcinoma (HCC). However, the mechanisms by which these 

infections alter host immunity are not well understood. In a 

previous RNA-Seq analysis, we identified Golgi reassembly 

stacking protein 2 (GORASP2) as a host factor that participates 

in IFN-a regulation of HCV replication (Lin et al, J Hepatology 

2015, 62:1024). GORASP2 is involved in establishing the 

Golgi apparatus structure. We hypothesized that GORASP2 

protein regulates HCV and HBV through HCV RNA replication, 

HBV DNA and pregenomic RNA replication, and HCV/ 

HBV virion trafficking. Methods: We performed siRNA knockdown 

or overexpression of GORASP2 in JFH1 HCV-infected 

Huh7.5.1 cells or HepG2.2.15 HBV replicon cells with or 

without IFN treatment. Selected gene mRNA variants and their 

proteins, together with HBV replication in cells and HBV virions 

in supernatant, were monitored by qRT-PCR and Western blot. 

HBV replication was assessed by measuring HBV X and S protein 

levels, HBV DNA levels and pregenomic RNA in cells, and 

supernatant virus DNA. Results: We found that GORASP2 has 

two isoforms based on RNA-Seq exon splicing bioinformatics 

analysis. GORASP2 V2 includes exon #2 while V1 does not. 

We found that siRNA knockdown of GORASP2 increased HCV 

RNA and protein levels in JFH1-infected Huh7.5.1 cells and 

HBV levels in DNA and pregenomic RNA in HepG2.2.15 cells. 

GORASP2 mRNA and protein expression were not affected by 

IFN treatment, indicating that GORASP2 expression is independent 

of direct IFN regulation. We found that GORASP2 

V1 overexpression reduced HCV RNA and protein levels in 

JFH1-infected cells and HBV DNA and pregenomic RNA levels 

in HepG2.2.15 cells. In contrast, overexpression of GORASP2 

V2 had no significant effect on HCV RNA or protein levels 

in JFH1 cells. Golgi phosphoprotein 3 (GOLPH3) has been


reported to regulate HCV virion secretion through interaction 

with phosphatidylinositol 4-phosphate. We confirmed that 

GOLPH3 knockdown reduced HCV virion secretion in JFH1 cell 

culture supernatants and HBV DNA and pregenomic RNA levels 

in HepG2.2.15 cells. Moreover, we found that GORASP2 

knockdown increased both intracellular HCV RNA replication 

and virion trafficking and release to the supernatant. Conclusions: 

We conclude that GORASP2 protein regulates HCV and 

HBV replication through RNA replication and virion trafficking. 

Further exploration of the mechanism by which GORASP2 or 

GOLPH3 alters viral nucleic acid replication and virion trafficking 

are warranted. 

Disclosures: 



Merck, BMS 

The following authors have nothing to disclose: Dachuan Cai, Jian Hong, Xiao 

Liu, Sae Hwan Lee, Esperance A. Schaefer, Cynthia Brisac, Anna Lidofsky, 

Wenyu Lin 

compared to those without sustained HBeAg seroconversion 

were shown in table. Multivariate analysis found pre-treatment 

HBVDNA < 7 logIU/ml (OR=5.7, 95%CI=1.1-28.6, p=0.03) 

and rs12794714 CYP2R1 TT genotype (OR=4.1, 95%CI=1.1- 

15.5, p=0.04) strongly predicted HBeAg seroconversion at 24 

weeks post treatment. HBVDNA before treatment and at week 

12 was significant lower in rs12794714 CYP2R1 TT genotype 

than non-TT genotype. There was no association between 

the remaining 12 SNPs and treatment response, including sustained 

HBeAg seroconversion, combined response (HBeAg 

loss and HBVDNA


of 1,653 Japanese samples representing 892 HBV patients 

and 761 healthy controls; we used the data from these 2,605 

individuals to assess the effects of HLA genotype on CHB infection. 

PATIENTS/METHODS: Of the 2,605 Japanese genomic 

DNA samples used in this study, 1,653 samples were newly 

obtained from healthy volunteers (n=761) or HBV patients 

(n=892 each patient was categorized as either inactive carrier 

(IC), chronic hepatitis (CH), or acute exacerbation of CHB, liver 

cirrhosis and hepatocellular carcinoma); the 892 patients were 

treated at 14 multi-center hospitals throughout Japan; the other 

952 patient samples were those used in our previous study. For 

each newly collected sample (n=1,653), HLA-DPB1 genotyping 

was performed by PCR-Luminex method. The associations of 

HLA-DPB1 alleles or genotypes with CHB infection or disease 

progression were assessed via a χ2 test. RESULTS: Significant 

associations between CHB infection and five DPB1 alleles (two 

susceptibility alleles, DPB1 * 05:01 and * 09:01, and three protective 

alleles, DPB1 * 02:01, * 04:01, and * 04:02) were confirmed 

in a population comprising 2,605 Japanese individuals. 

Odds ratios for CHB were higher for those with both DPB1 

susceptibility alleles than for those with only one susceptibility 

allele; therefore, effects of susceptibility alleles were additive 

for risk of CHB infection. Similarly, protective alleles showed an 

additive effect on protection from CHB infection. Moreover, heterozygotes 

of any protective allele showed stronger association 

with CHB than did homozygotes, suggesting that heterozygotes 

may bind a greater variety of hepatitis B-derived peptides, 

and thus present these peptides more efficiently to T cell receptors 

than homozygotes. Notably, compound-heterozygote of 

DPB1*04:02 (protective) and DPB1*05:01 (susceptibility) was 

significantly associated with protection against CHB infection, 

which indicated that one protective HLA-DPB1 molecule can 

provide dominant protection. CONCLUSIONS: Identification of 

the HLA-DPB1 genotypes associated with susceptibility to and 

protection from CHB infection is essential for future analysis of 

the mechanisms responsible for immune recognition of hepatitis 

B virus antigens by HLA-DPB1 molecules. 

Disclosures: 






Namiki Izumi - Speaking and Teaching: MSD Co., Chugai Co., Daiichi Sankyo 

Co., Bayer Co., Bristol Meyers Co. 

Naoya Sakamoto - Grant/Research Support: Gilead Sciences, TRSS; Speaking 

and Teaching: Bristol Myers Squibb, Janssen Pharm, Chugai co ltd 

The following authors have nothing to disclose: Nao Nishida, Jun Ohashi, Masaya 

Sugiyama, Takayo Tsuchiura, Ken Yamamoto, Keisuke Hino, Masao Honda, 

Hiroshi Yatsuhashi, Kazuhiko Koike, Osamu Yokosuka, Eiji Tanaka, Akinobu 

Taketomi, Masayuki Kurosaki, Yuichiro Eguchi, Takehiko Sasazuki, Katsushi 

Tokunaga, Masashi Mizokami, Tatsuya Kanto 

influence the outcome of the infection. Furthermore, SNPs in 

HLA-C have been implicated in hepatitis B chronicity. However, 

whether there is a role for these variations in prediction of 

treatment outcome in chronic hepatitis B (CHB) patients is currently 

unknown. Methods: We included 86 CHB patients (41 

HBeAg-positive; 45 HBeAg-negative) from a previously conducted 

study who completed 48 weeks of peginterferon alfa-2a 

and adefovir combination therapy followed by a treatment-free 

follow-up (week 72). Patient DNA was isolated from PBMCs. 

Twelve SNPs in the HLA-C gene (±500 base pairs) were genotyped 

with the Illumina Human Omni1-Quad BeadChip. Genotyping 

of KIRs was performed by PCR. Combined response at 

week 72 (HBeAg negativity, HBV-DNA ≤2,000 IU/mL, and 

ALT normalization) was achieved in 14/41 HBeAg-positive 

and 17/45 HBeAg-negative patients. Results: One SNP in 

HLA-C (rs2308557; A allele) was significantly associated with 

combined response in HBeAg-positive patients after correction 

for multiple testing (p=0.003), but not in HBeAg-negative 

patients. This SNP is in high linkage with the SNP defining the 

presence of an Asn or Lys at position 80 of the α 1 

domain of 

HLA-C, important for binding specific KIRs (HLA-C group C1 or 

C2, respectively). The distribution of C1C1, C1C2, C2C2 genotypes 

in all patients was 36 (42%), 36 (42%), and 17 (20%), 

respectively. HBeAg-positive patients with the combination 

KIR2DL1-C2 had higher rates of combined response (13/24 

vs 1/17, p=0.001), and higher baseline ALT levels (median 

ALT 136 vs 50 U/L, p=0.002) compared to patients with 

only C1 alleles. In HBeAg negative patients, the combination 

KIR2DS2-C1 was more frequent in combined responders versus 

non-responders (11/17 versus 9/28 respectively, p=0.037) In 

multivariable analysis, presence of HLA-C2/KIR2DL1 predicted 

response independent of HBV genotype A (p=0.012) and ALT 

levels at baseline (p=0.005). Furthermore, the presence of HLA- 

C1/KIR2DS2 predicted response in HBeAg negative patients 

independent of HBV DNA (p=0.045) and HBsAg levels at 

baseline (p=0.040). Conclusion: KIR/HLA-C combination was 

strongly associated with combined response in both HBeAg 

positive and negative CHB patients treated with combination 

therapy. These findings support an important role for the interaction 

of NK cell receptors and their HLA-C ligands in determining 

host immune activity and response to interferon-based 

therapy in chronic hepatitis B. 

Disclosures: 

Bart Takkenberg - Speaking and Teaching: Roche, Gilead, BMS 

Hendrik W. Reesink - Consulting: Abbvie, Alnylam, BMS, Gilead, Janssen-Cilag, 

Merck, PRA-International, Regulus, Replicor, Roche, R-Pharm; Grant/Research 

Support: AbbVie, BMS, Boehringer Ingelheim, Gilead, Janssen-Cilag, Merck, 

PRA-International, Regulus, Replicor, Roche 

The following authors have nothing to disclose: Femke Stelma, Louis Jansen, 

Marjan J. Sinnige, Karel A. van Dort, Ester M. van Leeuwen, Neeltje A. Kootstra 

1633 

The Combination of KIR and HLA-C Is Associated with 

Response to Peginterferon and Adefovir Combination 

Therapy in Chronic Hepatitis B Patients. 

Femke Stelma 1,2 , Louis Jansen 1,2 , Marjan J. Sinnige 2 , Karel A. van 

Dort 2 , Bart Takkenberg 1 , Ester M. van Leeuwen 2 , Neeltje A. Kootstra 

2 , Hendrik W. Reesink 1 ; 1 Gastroenterology and Hepatology, 

Academic Medical Center, Amsterdam, Netherlands; 2 Experimental 

Immunology, Academic Medical Center, Amsterdam, Netherlands 

Introduction: Killer cell immunoglobulin-like receptors (KIR) 

expressed on NK cells interact with human leukocyte antigen 

class C (HLA-C) ligands. From acute hepatitis C studies, it is 

known that genetic variations in KIR/HLA-C combinations can 

1634 

NFATc3 exerts anti-viral and anti-tumor dual function 

in hepatitis B virus-related hepatocellular carcinoma by 

activating interferon pathway 

Xiangmei Chen, Jin Cheng, Zhenzhen Zeng, Fengmin Lu; Department 

of Microbiology & Infectious Disease Center, Peking Univercity 

Health Science Center, Beijing, China 

Purpose: This study aimed to explore the aberrant status of 

innate immune response-related genes, NFATc3 and ADAR1, 

and their potential function in HBV-related hepatocellular carcinoma(HCC). 

Methods: Real-time PCR was used to determine 

the mRNA level of NFATc3 and ADAR1 in HCC samples. The 

transcriptional regulation of IFN-related genes by NFATc3 or 

ADAR1 was detected by luciferase reporter assay and realtime 

PCR. The anti-HBV function of NFATc3 was evaluated


in vitro andin vivo. CCK-8 assay, wound healing, and transwell 

assay were used to determine the effect of NFATc3 

on HCC oncogenic properties. Results: Our previous aCGH 

results showed that NFATc3 gene was frequently deleted in 

HCC samples(7/24), while ADAR1 gene was largely amplified(10/24). 

NFATc3 was significantly downregulated in HCC 

tissues compared to paired non-tumor tissues(P=0.002, N=75), 

and the decrease of NFATc3 in tumor tissues was significantly 

correlated with poor prognosis(P=0.021). ADAR1 was significantly 

upregulated in HCC tissues(P=0.007, N=66). NFATc3 

overexpression induced IFN-α, β and γ transcriptional activity 

and upregulated the mRNA level of IFN stimulated genes 

in HCC cells, while ADAR1 suppressed the transcriptional 

activity of IFN-β promoter. After mutating of NFAT binding 

site within the IFN-β gene promoter, NFATc3 failed to induce 

its transcriptional activity, demonstrating that NFATc3 exerted 

its function through directly binding with IFN gene promoter. 

In Hep3B cells transiently transfected with 1.2x HBV replicon 

or HepAD38 cells stably expressing HBV, NFATc3 overexpression 

significantly decreased HBsAg, HBeAg, HBV pregenomic 

RNA and HBV total DNA level, and adding IFN receptor 

blocker in the culture medium abolished this inhibitory effect. 

While in immune deficient HCC cell line Huh-7, ectopic expression 

of NFATc3 couldn’t activate IFN pathway or suppress HBV 

replication. The above observations demonstrated that NFATc3 

exerted its anti-HBV function by upregulating IFN pathway. In 

mouse model established by hydrodynamic tail vein injection of 

1.2x HBV replicon, co-injection of NFATc3 expression plasmid 

significantly decreased HBsAg and HBeAg level in peripheral 

blood of the mice. More importantly, for HCC patients with 

lower NFATc3 expression, the HBV DNA level in their peripheral 

blood was significantly higher than patients with higher 

NFATc3(P=0.040). Lastly, restoration of NFATc3 significantly 

reduced the proliferation and migration ability of HCC cells. 

Conclusion: Our results suggest that NFATc3 exerts anti-viral 

and anti-tumor dual function by activating IFN pathway, which 

provided a novel option of clinical treatment for HBV-related 

HCC by targeting NFATc3. 

Disclosures: 

The following authors have nothing to disclose: Xiangmei Chen, Jin Cheng, 

Zhenzhen Zeng, Fengmin Lu 

1635 

Generation of human peripheral blood mononuclear 

cells-engrafted mice for the study of immunological 

response against hepatitis B virus 

Satoshi Aono 1 , Tomohide Tatsumi 1 , Seiichi Tawara 1 , Yoshiki Onishi 

1 , Akira Nishio 1 , Shigeki Suemura 1 , Hayato Hikita 1 , Ryotaro 

Sakamori 1 , Naoki Hiramatsu 1 , Hiroshi Suemizu 2 , Takeshi Takahashi 

2 , Tetsuo Takehara 1 ; 1 Osaka University, Suita, Japan; 2 Central 

Institute for Experimental Animals, Kawasaki, Japan 

Background&Aims: Immunologically humanized mouse model 

is required for the assessment of immunological responses 

against hepatitis B virus (HBV) . Human peripheral blood mononuclear 

cells (PBMC) –engrafted mouse model using immunodificient 

NOG mice resulted in severe graft-versus-host disease. 

Recently, we generated NOG-Iaβ/β2m double KO mice which 

were NOG mice deficient in both MHC class I and II (DKO- 

NOG mice) . In this study, we established a new humanized 

mouse model capable of evaluating immune response against 

HBV derived proteins. Methods and Results: We used NOG 

(NODShi.Cg-Prkdc scid Il12rg tm1sug ) mice and DKO-NOG mice. 

Human PBMC were inoculated into these mice via tail vein. 

After injection of human immune cells, both infiltration into the 

liver of human immune cells and hepatocyte damage were 

observed in NOG mice, but not in DKO-NOG mice. Serum 

ALT levels were severely elevated in NOG mice, but not in 

DKO-NOG mice (510±299 IU/l vs, 14.5±4.12 IU/L at day 

15, 939±444 IU/l vs, 36.4±19.7 IU/L at day 29, respectively) 

. Seven of 8 NOG mice died within 2 months after injection of 

human PBMC whereas all DKO-NOG mice survived more than 

70 days. Liver mononuclear cells (MNCs) isolated from these 

mice were subjected to flow cytometry. On day 28 after injection 

of human PBMC, replacement rates of human immune cells 

in the liver increased up to 85% in DKO-NOG mice. On day 

15, the expressions of PD-1 and Tim-3 on T cells from DKO- 

NOG mice were significantly lower than those from NOG 

mice. On day 15, the frequencies of B cells and dendritic cells 

(DCs) in DKO-NOG mice were significantly higher than those 

in NOG mice. Next, we evaluated the production of anti-HBs 

antibody (anti-HBs) in sera of human PBMC-engrafted DKO- 

NOG mice. Inoculations of recombinant hepatitis B vaccine 

resulted in the production of anti-HBs in 2 of four vaccinated 

mice and also in the increase of CD138, a plasma cell marker, 

positive B cells in splenocytes. Finally, We evaluated the induction 

of HBc-derived peptide-specific cytotoxic T lymphocytes 

(CTLs) by using specific tetramer in human PBMC-engrafted 

DKO-NOG mice. Vaccination of HBc-derived peptide-pulsed 

DCs resulted in significant increase of HBc-derived peptide-specific 

CTLs in vaccinated mice. Moreover, HBV expression in the 

liver by hydrodynamic injection resulted in significant increase 

of HBc-derived peptide-specific CTLs. Conclusion: The present 

study demonstrates that immunological responses against HBV 

could be induced in the human PBMC-engrafted DKO-NOG 

mice. This system would provide us with a new promising 

model evaluating immunological responses against HBV in 

human chimeric livers. 

Disclosures: 






The following authors have nothing to disclose: Satoshi Aono, Tomohide Tatsumi, 

Seiichi Tawara, Yoshiki Onishi, Akira Nishio, Shigeki Suemura, Ryotaro Sakamori, 

Naoki Hiramatsu, Hiroshi Suemizu, Takeshi Takahashi 

1636 

NK cells play important roles in the pathogenesis of 

HBV infection via alternation of NKp46 high NKG2A high 

subset 

Teppei Yoshioka, Yoshinobu Yokoyama, Akira Nishio, Yoshiki 

Onishi, Kaori Mukai, Satoshi Aono, Minoru Shigekawa, Hayato 

Hikita, Ryotaro Sakamori, Naoki Hiramatsu, Takuya Miyagi, 

Tomohide Tatsumi, Tetsuo Takehara; Gastroenterology and Hepatology, 

Osaka university graduate school of medicine, Suita, Japan 

Background/Aim: Natural Killer (NK) cells attract attention in 

the control of viral infection. However, the role of NK cells 

against HBV infection remains controversial. The aim of this 

study was to investigate the role of NK cells in the pathogenesis 

of chronic hepatitis B (CHB), focusing on an activating receptor 

NKp46 and an inhibitory receptor NKG2A. Method: Untreated 

CHB patients (n = 53) and healthy subjects (HS; n = 35) were 

enrolled. Peripheral blood lymphocytes were obtained and 

the frequencies of NKp46 and NKG2A were analyzed by 

flow cytometry. To evaluate the function in each NK subsets, 

we examined the cytotoxicity by RT-PCR and flow cytometry. 

NK cells were co-cultured with HepG2.2.15 (2215), which 

express HBV related protein, or HepG2 (G2), and the role of 

NKp46 was evaluated by neutralizing antibody of NKp46 

(9E2). Finally, alternation of the frequencies of NK cell subset


in patients treated with nucleos(t)ide analogue (NA; n = 17) or 

interferon-α (IFNα; n = 9) were analyzed by flow cytometry. 

Result: CHB patients were divided into inactive CHB group 

(I-CHB, n = 27; patients showed both serum ALT less than 40 

IU/ml and HBV-DNA less than 4 LC /ml) and active CHB group 

(A-CHB, n = 26; remaining patients). The expression of NKp46 

was higher in A-CHB than in HS or I-CHB. A unique subset 

which strongly expressed both NKp46 and NKG2A was identified 

(NKp46 high NKG2A high subset). The frequencies of this 

subset in NK cells were 4.8 ± 2.9%, 4.9 ± 3.6% and 10.3 ± 

6.3% in HS, I-CHB and A-CHB, respectively. The frequencies of 

this subset positively correlated with serum ALT level (P


using Chi-square test and logistic regression analysis. Results: 

In the CHB group, 28.5% were inactive carriers, 67.7% were 

HBe-antigen negative, 17.2% had cirrhosis, 6.5% had hepatocellular 

carcinoma and 4.4% were liver transplanted. 85.6% 

had already received treatment for CHB. The mean age and 

sex distribution differed significantly between the CHB and the 

SC groups (45.38±15.4 vs. 61.35±14.5; p


HBV-specific CD8+ T cells was almost completely diminished in 

the absence of MHC class I expression on non-hematopoietic 

cells, resulting in more than 2,000 fold decrease in the number 

of HBV-specific CD8 T cells compared with their counterparts 

in MHC class I positive control animals. HBV-specific CD8+ 

T cells did not express granzyme B, either, even though they 

clearly recognized antigen and expressed activation marker 

CD69. Conclusion: These results suggest that cross-presentation 

by hematopoietic cells is required for the optimum expansion 

of HBV-specific CD8 T cells, but endogenous antigen presentation 

by non-hematopoietic cells, presumably hepatocytes, is far 

more essential in the expansion and cytolytic differentiation of 

HBV-specific CD8+ T cells. 

Disclosures: 



Squibb 

The following authors have nothing to disclose: Masanori Isogawa, Yasuhiro 

Murata, Keigo Kawashima 

1641 

Initial sites of hepadnavirus-host genome integration 

after de novo infection of HepaRG cells and woodchuck 

livers 

Ranjit Chauhan, Norma D. Churchill, Thomas I. Michalak; Molecular 

Virology and Hepatology Research Group, Memorial University, 

St. John’s, NF, Canada 

Backgound: HBV is highly oncogenic virus and its integration 

into human genome precedes development of hepatocellular 

carcinoma (HCC). It is expected that the sites and the spread 

of HBV integrations across human genome might be relevant to 

the establishment and progression of HCC. The initial sites of 

HBV-host DNA junctions remain unknown. Aim: To identify the 

earliest sites of hepadnavirus integration into host’s genome by 

exploring de novo infection of human hepatocyte-like HepaRG 

cells with authentic HBV and woodchucks infected with woodchuck 

hepatitis virus (WHV). Methods: HepaRG cells infected 

with HBV were investigated in 15 min, 30 min, 1 h, 3 h, 24 

h, 72 h, 7 days (d), 10 d, 2 weeks (wk), 4 wk and 7 wk 

post-infection. Liver biopsies were obtained from woodchucks 

infected with WHV at 1 h or 3 h and at 6 wk post-infection (JVI 

2008;82:8579). HepaRG cells were analyzed for HBV X gene 

integrations, while liver biopsies for WHV X and preS sequence 

integrations applying inverse-PCR followed by amplicon cloning 

and sequencing. Results: Intracellular HBV DNA and its replication 

intermediates became detectable in HepaRG cells from 

1 h and remained present up to 7 wk post-exposure. HBV DNA 

integrations became identifiable in 1 h post-infection. Junctions 

of HBV X gene in retrotransposon sequences, such as LINE1 

(long-interspersed nuclear element 1) and LINE2, became 

prominent from 3 days post-infection and they represented 

approximately half the virus-host junctions detected at that time. 

Insertions of HBV X integrant into other genes, including human 

satellite HSAT-II sequence, neurotrimin on chromosome (Ch) 

11q25, ZNF-782 on Ch-9q22.3, myosin 3B on Ch-2q31.1, 

and ten-eleven translocation-1 gene (TET-1) on Ch-10q21.3 

were also detected. In woodchucks, integration of WHV preS 

genomic sequence into MAML2 gene on Ch-11q21 (based on 

88% homology with human) was identified in 1 h post-infection 

and WHV X gene integrants into unidentified so far woodchuck 

genes were evident in 1 h and 3 h post-infection. The 

HBV core promoter/enhancer-II region followed by enhancer I 

region and its WHV equivalent were predisposed to form the 

earliest junctions with host DNA. In addition, multiple virus-virus 

DNA joints were identified from 1 h post-infection onward in 

both HBV-infected cells and WHV-infected woodchuck livers. 

Conclusions: HBV DNA integrates very early into the host’s 

genome and may utilize retrotransposon elements and translocation 

genes from the early stages of infection for spread and 

induction of pro-oncogenic perturbations throughout the host’s 

genome. The predisposition to very early integration was also 

evident in natural WHV infection in woodchucks. 

Disclosures: 

The following authors have nothing to disclose: Ranjit Chauhan, Norma D. Churchill, 

Thomas I. Michalak 

1642 

An impact of cellular core-fucosylation in hepatitis B 

virus infection into hepatoma cell lines 

Eiji Miyoshi 2 , Shinji Takamatsu 1 , Mayuka Shimomura 1 , Yoshihiro 

Kamada 1,3 , Haruka Maeda 1 , Tomoaki Subajima 2 , Hayato Hikita 3 , 

Iijima Mayumi 4 , Yuta Okamato 5 , Ryo Misaki 5 , Kazuhito Fujiyama 5 , 

Tetsuo Takehara 3 , Keiji Ueda 6 , Shunichi Kuroda 4 ; 1 Department of 

Molecular Biochemistry & Clinical Investigation, Osaka University 

Graduate School of Medicine, Suita, Japan; 2 Department of 

Molecular Biochemistry and Clinical Investigation, Osaka University 

Graduated School of Medicine, Suita, Japan; 3 Department of 

Gastroenterology and Hepatology, Osaka University Graduated 

School of Medicine, Suita, Japan; 4 Department of Bioengineering 

Sciences, Graduate School of Bioagricultural Sciences, Nagoya 

University, Nagoya, Japan; 5 Applied Microbiology Laboratory, 

International Center for Biotechnology, Osaka University, Suita, 

Japan; 6 Department of Microbiology, Osaka University Graduated 

School of Medicine, Suita, Japan 

Background & Aims: It is a well known fact that oligosaccharides 

on cell surface proteins such as growth factor receptors 

as well as virus/bacteria entry receptors regulate their functions 

dynamically. These oligosaccharide structures are regulated 

by a set of expression of several kinds of glycosyltransferase. 

In the present study, we investigated the involvement of glycosylation 

in hepatitis B virus (HBV) entry onto hepatoma cells. 

Methods: We used several kinds of hepatoma cell lines such 

as Huh7, Hep3B, Huh6 and HB611 cells. HB611 cells are 

established by transfection of tandem repeats of HBV genome 

DNA into Huh6 cells and secrete HBV-related antigens into 

the conditioned medium. Bio nanocapsules (BNC) are hollow 

particles consisting of HBV surface antigen (HBs) large proteins 

embedded in a unilamellar liposome, and used as pseudo-HBV 

virus. Incorporation of BNC in oligosaccharide remodeling 

hepatoma cells was evaluated by flow cytometer and con-focal 

microscopy. Oligosaccharide structure was analyzed with 

lectin blot, lectin microarray and mass spectrometry. To identify 

association molecules with sodium taurocholate cotransporting 

polypeptide (NTCP), which is one of major candidate for HBV 

receptors, immune-precipitation with NTCP antibody followed 

by LC-MS analyses was performed. Results: Transfection of 

glycosyltransferase genes into several kinds of hepatoma cells 

changed the level of BNC uptake. Among them, HB611 cells 

showed dramatic increases in BNC incorporation, compared 

to parental cell, Huh6 cells. As a result from glycomic analyses, 

increases in cellular fucosylation and sialylation were observed 

in HB611 cells. Knock down of fucosyltransferase 8 (FUT8) 

gene as well as sialidase treatment decreased BNC entry into 

HB611 cells. In contrast, overexpression of FUT8 in Huh6 cells 

increased BNC incorporation. Addition of core-fucose specific 

lectin, Pholiota squarrosa lectin (PhoSL) in the conditioned 

medium inhibited the entry of BNC into HB611 cells. While 

expression of sodium taurocholate cotransporting polypeptide 

(NTCP) was lower in HB611 cells than Huh6 cells, levels of 

co-precipitated molecules with NTCP were dependent on levels 

of core fucosylation, suggesting that core-fucosylation regu-


lates BNC entry into hepatoma cells. As a result from confocal 

microscopy observation, endocytosis of NTCP seemed to be 

up-regulated in HB611 cells in terms of core fucosylation. Conclusions: 

Core-fucosylation and sialylation have pivotal roles in 

HBV infection into hepatoma cells. Especially, downregulation 

of core-fucosylation with Fut8 knockdown or treatment with 

core-fucose specific lectin might be a novel target for HBV 

therapy. 

Disclosures: 






The following authors have nothing to disclose: Eiji Miyoshi, Shinji Takamatsu, 

Mayuka Shimomura, Yoshihiro Kamada, Haruka Maeda, Tomoaki Subajima, 

Iijima Mayumi, Yuta Okamato, Ryo Misaki, Kazuhito Fujiyama, Keiji Ueda, 

Shunichi Kuroda 

1643 

Indoleamine-2, 3-dioxygenase as the intrinsic anti-HBV 

effector that leads to durable immune responses in 

patients with acute hepatitis B: A comparative analysis 

with the sequence in hepatic flare 

Sachiyo Yoshio 1 , Masaya Sugiyama 1 , Hirotaka Shoji 1 , Yohei 

Mano 1 , Yoshihiko Aoki 1 , Toru Okamoto 2 , Yoshiharu Matsuura 2 , 

Masashi Mizokami 1 , Tatsuya Kanto 1 ; 1 The Research Center for 

hepatitis and immunology, National Center for global health and 

medicine, Chiba, Japan; 2 Molecular Virology, Research Institute 

for Microbial Diseases, Osaka University, Suita, Japan 

BACKGROUD&AIMS: Primary HBV infection to immune competent 

adults is mostly a self-limited disease, resulting in the HBV 

clearance. However, HBV is hard-to-eradicate once it attains 

persistent infection. The factors that distinguish these conditions 

should be crucial for the control of HBV. Indoleamine-2, 3-dioxygenase 

(IDO) is induced in hepatocytes by IFN-γ and catalyzes 

tryptophan to form kynurenine. IDO exerts dual function 

in infectious diseases; acting as a suppressor of intracellular 

pathogens and as an immune regulator. We aimed to explore 

the factors, including IDO, capable of suppressing HBV, by 

comprehensive immunological analysis between patients with 

acute hepatitis with HBV clearance and those at hepatic flare 

with persistent infection. METHODS: We enrolled HBV-positive 

patients with acute hepatitis (AH, N=26), chronic hepatitis 

(CH, 12), hepatic flare (HF, 14) and healthy volunteers (HV, 

10) in this study. We longitudinally compared serum levels of 

40 cytokines and chemokine by a multiplexed assay. Systemic 

IDO activity was determined by the ratio of kynurenine to tryptophan 

measured by HPLC. In order to examine the mechanism 

of IDO induction and its impact on HBV replication, we set up 

the co-culture consisting of HBV transfected Huh7 (HBV+Huh7), 

freshly-isolated human NK cells and plasmacytoid dendritic 

cells (pDCs). The IDO-knock out (IDO-KO) Huh7 was established 

by CRISPR-Cas9 system. RESULTS: The AH group showed 

a robust increase of IDO activity prior to the peak of ALT elevation, 

the levels of which were higher than those in the CH, 

HF or HV group. The IDO activity subsided overtime in accordance 

with the decline of ALT and HBVDNA. In the AH group, 

the levels of CXCL9, CXCL10 and CXCL11 were increased in 

the acute phase, followed by the sustained increase of IFN-γ, 

TNF-α, CCL19 and CCL22 until the disappearance of HBsAg. 

In contrast, such sequential transition of the immune factors, 

beginning from the robust IDO activation, was not observed 

in the HF group. In the co-culture, the coexistence of NK and 

pDCs synergistically increased the production of IFN-γ and 

IFN-α thereby enhancing IDO activity in HBV+Huh7, resulting 

in HBV suppression. In IDO-KO HBV+Huh7, the suppressive 

effect of NK and pDCs on HBV replication was abrogated 

(P


MX1 N-terminal domain to specifically identify its effects on 

HBV and HCV replication is warranted. 

Disclosures: 



Merck, BMS 

The following authors have nothing to disclose: Wenyu Lin, Dachuan Cai, Jian 

Hong, Xiao Liu, Hong Zhao, Chuanlong Zhu, Esperance A. Schaefer, Cynthia 

Brisac, Sae Hwan Lee, Anna Lidofsky 

Disclosures: 








The following authors have nothing to disclose: Masataka Tsuge, Nobuhiko 

Hiraga, Hiromi Abe, Daiki Miki, Michio Imamura, Hidenori Ochi, C. Nelson 

Hayes 

1645 

Difference of intracellular responses by HBV genotype A 

and C infection in humanized mouse model 

Masataka Tsuge 1 , Nobuhiko Hiraga 1 , Hiromi Abe 1 , Daiki Miki 2 , 

Michio Imamura 1 , Hidenori Ochi 1 , C. Nelson Hayes 1 , Kazuaki 

Chayama 1 ; 1 Department of Gastroenterology and Metabolism, 

Hiroshima university, Hiroshima, Japan; 2 Institute of Physical and 

Chemical Research (RIKEN), Hiroshima, Japan 

Background & Aims: It is well known that the characteristics of 

chronic hepatitis B, including the clinical course and responsiveness 

to anti-viral treatments, differ among hepatitis B virus 

(HBV) genotypes. However, the cause of these differences has 

not been clarified. We have previously established a chronic 

HBV infection model using human hepatocyte transplanted 

uPA-SCID mice lacking human immune cells. We previously 

reported the effects of HBV infection in human hepatocytes 

using cDNA microarray. In the present study, we analyzed 

mRNA expression profiles using next generation sequencing in 

this mouse model and compared the responses between HBV 

genotypes A and C. Methods: Fifteen chimeric mice were prepared 

and divided into the following three groups: uninfected 

control mice, HBV genotype A infection, and HBV genotype C 

infection. Ten mice were inoculated with HBV genotype A or C, 

and human hepatocytes in the mouse livers were collected after 

mouse serum HBV DNA titers had reached a plateau of around 

8 Log copies/ml. To analyze changes in gene expression in 

human hepatocytes, we performed next generation sequencing 

and compared gene expression profiles. Results: HBV genotype 

A and C infection resulted in significant changes in mRNA 

expression levels in HBV-infected hepatocytes in 780 and 208 

genes, respectively (P


B, where the liver damage is T-cell mediated, our data suggest 

that a unique type of humoral immunity against HBcAg plays a 

primary role in the pathogenesis of HBV-ALF. 

Disclosures: 

The following authors have nothing to disclose: Zhaochun Chen, Huaying Zhao, 

Peter Schuck, Ronald E. Engle, Fausto Zamboni, Giacomo Diaz, David E. Kleiner, 

Robert H. Purcell, Patrizia Farci 

1647 

The expansion of CD11b - CD27 - NK-cell subset contributes 

to HBV persistence via TGF-β1 

Qiongfang Zhang 1,2 , Wenwei Yin 1,2 , Jian-ying Shao 1,2 , Qian 

Liu 1,2 , Peng Hu 1,2 , Huaidong Hu 1,2 , Hong Ren 1,2 , Dazhi Zhang 1,2 ; 

1 The institute of viral hepatitis,Department of infectiouse diseases, 

Chongqing Medical University, Chongqing, China; 2 The Second 

Affiliated Hospital of Chongqing Medical University, Chong qing, 

China 

Background /purposeThe mechanism underlying persistent 

hepatitis B virus (HBV) infection remains unclear. Natural killer 

(NK) cells, as a major component of the antiviral immune 

response, has been reported to alter their distribution of NK-cell 

subsets. We reported a novel mechanism associated with viral 

persistence from NK cell-mediated antiviral immunity in patients 

in immune tolerant (IT) phase. Method We investigated the 

role of NK subsets to persistent HBV infection in 109 chronic 

hepatitis B (CHB) patients and 38 healthy controls (HC) by flow 

cytometry. Serum cytokine concentrations were analyzed using 

a Cytometric Bead Array (CBA) Inflammation Kit and a standard 

sandwich enzyme-linked immunosorbent assay (ELISA) 

Kit. Result We found a substantial CD11b - CD27 - (DN) NK-cell 

population harboured in patients in IT phase compared with 

patients in immune active (IA) phase, patients in immune inactive 

(IN) phase and HC. And there existed a positive correlation 

between the expanding DN NK-cell subtype and HBV-DNA 

load. This DN NK subset displayed an immature and inactive 

phenotype and showed poor degranulation and IFN-γproduction. 

Higher concentrations of transforming growth factor beta1 

(TGF-β1) were found in sera from IT-phase patients, which positively 

correlated with HBV-DNA load. In vitro, TGF-β1 up-regulated 

the presence of DN NK-cell population. Moreoverthe, 

percentage of DN NK-cell subset in fresh NK cells preincubated 

with sera from IT patients was markedly higher compared with 

IA phase patients, IN phase patients and HC. Anti-TGF-β1 antibodies 

could partially restore the altered NK-cell subsets. Conclusion 

Our findings demonstrated that the IT microenvironment 

may render circulating NK cells into an inefficient subtype by 

up-regulating the levels of TGF-β1 in serum, which was correlated 

with persistent HBV infection. 

Disclosures: 

The following authors have nothing to disclose: Qiongfang Zhang, Wenwei Yin, 

Jian-ying Shao, Qian Liu, Peng Hu, Huaidong Hu, Hong Ren, Dazhi Zhang 

1648 

Comprehensive analysis of the Rab family to screen 

membrane traffic pathways utilized by hepatitis B virus 

Jun Inoue, Yasuteru Kondo, Teruyuki Umetsu, Takuya Nakamura, 

Takayuki Kogure, Yu Nakagome, Yuta Wakui, Tatsuki Morosawa, 

Yasuyuki Fujisaka, Satoshi Takai, Tooru Shimosegawa; Division 

of Gastroenterology, Tohoku University Graduate School of Medicine, 

Sendai, Japan 

Background/aim: The trafficking pathways of hepatitis B virus 

(HBV) components in the host cells have remained largely 

unknown. Previously, we showed that the small GTPase Rab7 

regulates HBV secretion from multivesicular bodies (MVBs) 

by facilitating viral degradation by lysosomes (Inoue at al., 

J Cell Sci 2015). It is thought that the HBV core particles are 

enveloped in MVBs, but the transport pathway of the envelope 

HBs proteins to MVBs has not been determined. In this study, 

we performed a comprehensive screening of Rab proteins to 

identify important pathways in the assembly/release of HBV. 

Methods: Using HepG2.2.15 cells stably expressing HBV, 61 

Rab proteins were depleted separately with siRNA-mediated 

knockdown. With an siRNA concentration that maintained cell 

viability of more than 80%, HBV DNA and HBsAg in the culture 

supernatant were quantified 3 days after siRNA transfection. 

Results: The effects of Rab knockdown were divided into 7 

groups as follows: 1) decreased HBV DNA, 2) increased HBV 

DNA, 3) decreased HBsAg, 4) increased HBsAg, 5) decreased 

both HBV DNA and HBsAg, 6) increased both HBV DNA and 

HBsAg, 7) increased HBV DNA but decreased HBsAg. Group 

5 Rab proteins, which were considered to play roles in the 

assembly/release of both HBsAg subviral particles (SVPs) 

and HBV particles included Rab8, Rab13, and Rab38. These 

were reported to be associated with the vesicle transport from 

Golgi. Interestingly, the depletion of Rab5B, which is one of 

three isotypes of Rab5 and is known to be associated with 

early endosomes, increased both HBV DNA and HBsAg. The 

Rab5B depletion increased HBV DNA significantly more than 

the depletion of other Rab5 isotypes and the effect was rescued 

by the expression of GFP-Rab5B. Sucrose density gradient centrifugation 

revealed that the density of increased HBV particles 

after the Rab5B depletion was lowered from 1.24 g/ml to 

1.17-1.19 g/ml. These data indicate that Rab5B depletion 

may increase the release of immature HBV particles whose 

envelope components are altered. The significance of Golgi-related 

Rab proteins and Rab5B in early endosomes for the 

formation of mature HBV particles suggests the presence of a 

novel HBV life cycle pathway: HBs proteins are transported to 

the cellular membrane once via Golgi, and are endocytosed to 

early endosomes and late endosomes/MVBs, where the HBV 

core particles are thought to be enveloped. Conclusion: The 

findings of the Rab family screening suggested the importance 

of both the Golgi-related pathway and endocytotic pathway in 

the formation of mature HBV particles. This is a novel concept 

in the HBV life cycle and these steps could become targets of 

antiviral therapies. 

Disclosures: 

The following authors have nothing to disclose: Jun Inoue, Yasuteru Kondo, 

Teruyuki Umetsu, Takuya Nakamura, Takayuki Kogure, Yu Nakagome, Yuta 

Wakui, Tatsuki Morosawa, Yasuyuki Fujisaka, Satoshi Takai, Tooru Shimosegawa 

1649 

Genome-wide scan of miRNA polymorphisms revealed 

a suggestive association of miR-3143 with chronic hepatitis 

B virus infection 









Metabolism, Hiroshima University Hostital, Hiroshima, Japan 

Background & Aims: Recent genome-wide association studies 

(GWAS) have revealed several host genetic factors associated 

with various liver diseases, but the majority of genetic factors 

remain unclear. MicroRNAs (miRNAs) are small non-coding 

RNAs that control gene expression post-transcriptionally and


may explain some of the missing heritability. Single nucleotide 

polymorphisms in mature or precursor miRNAs (miRNA-SNPs) 

could alter expression levels or binding-affinity to their target 

genes, resulting in changes of expression levels of many target 

genes. In this study, we conducted a large scale case-control 

study of chronic hepatitis B virus (HBV) infection focusing 

on miRNA-SNPs. Methods: We extracted autosomal SNPs 

in mature or precursor miRNA sequences using dbSNP137 

and miRBase19 and excluded indel variations and miR-SNPs 

with minor allele frequencies (AFs) < 1% in the Asian population, 

as reported by the 1000 Genomes database. These 

miR-SNPs were then genotyped in 2,564 chronic HBV patients 

and 1,128 healthy controls by multiplex-PCR-based Invader 

assay. Results: Based on a database search, we selected 315 

autosomal miR-SNPs with minor AFs > 1%, of which 93 were 

in mature miRNAs and the remaining 222 were in precursor 

miRNAs. We successfully genotyped 205 miR-SNPs and analyzed 

them using an additive model for genotype-phenotype 

association. We detected only one SNP that reached significance 

after adjustment for multiple testing (P = 7.7 × 10 -6 

< 0.05/205 = 2.4 × 10 -4 ). This SNP, rs17737028, located 

in the precursor of miR-3143, showed a significantly higher 

minor AF in HBV patients than in healthy controls (0.048 and 

0.027, respectively) with odds ratio 1.81. Because this miR- 

SNP was located on chromosome 6 and was not far from 

the MHC region containing HLA-DPs previously reported by 

GWAS as the locus most associated with chronic HBV infection, 

we decided to examine whether our result were affected 

by HLA-DP genotype. After adjustment for rs3077 in HLA-DPA1 

and rs9277535 in HLA-DPB1, we still found significant association 

between rs17737028 in pre-miR-3143 and chronic 

HBV infection, although the significance level was substantially 

reduced (P = 0.027, OR = 1.42) despite weak linkage 

between this miR-SNP and the 2 HLA-DP SNPs. Conclusions: 

We performed a genome-wide scan of miRNA polymorphisms 

and found that rs17737028 in pre-miR-3143 was significantly 

associated with chronic HBV infection, although the association 

might be affected considerably by HLA-DP genotype. Further 

validation studies and functional analyses of miR-3143 as well 

as its target genes are needed. 

Disclosures: 













1650 

Hepatitis delta virus-induced T-cell recruiting chemokines 

contribute to inflammatory liver injury in chronic 

hepatitis delta 

Eui-Cheol Shin 1,2 , Nydiaris Hernandez-Santos 1 , Christopher Koh 1 , 

Mary DeMino 1 , Theo Heller 1 , Barbara Rehermann 1 ; 1 Liver Diseases 

Branch, NIDDK, National Institutes of Health, Bethesda, MD; 

2 Korea Advanced Institute of Science and Technology, Daejeon, 


Hepatitis delta virus (HDV) is a small defective RNA virus that 

depends on hepatitis B virus (HBV) surface antigen to envelope 

its genome. Superinfection of chronic hepatitis B with 

HDV severely enhances inflammatory liver injury and progression 

to liver cirrhosis. The reason for the enhanced disease 

pathogenesis remains unknown. Here, we compared HDV 

and HBV-specific T cell and chemokine responses of 16 HDV/ 

HBV co-infected patients to those of 25 HBV-infected patients. 

HDV/HBV co-infected patients had significantly higher ALT 

values than HBV-infected patients (medium 92 vs. 36 IU/L, 

p=0.001). Using IFN-gamma Elispot assays with overlapping 

HDV peptides we detected strong HDV-specific CD4 and CD8 

T cell responses in the peripheral blood of a patient with acute 

HDV/HBV infection and a patient who spontaneously cleared 

chronic HDV infection. However, HDV-specific T cell responses 

were barely detectable in chronic HDV/HBV co-infection, and 

were not unmasked by depletion of regulatory T cells or generation 

of T cell lines. Likewise, HBcore and HBsurface-specific 

CD4 and CD8 T cell responses were weak in HDV/HBV co-infected 

patients and did not differ from those of HBV-infected 

patients. Next, we quantitated serum chemokines known to 

recruit T cells to inflammatory sites. The serum levels of the 

two CXCR3-ligands CXCL9 and CXCL10, but not the chemokines 

CCL2, CCL3, CCL4 and CCL5 were significantly higher 

in HDV/HBV co-infected patients than in HBV-mono-infected 

patients. Serum CXCL9 and CXL10 levels correlated with ALT 

levels and histological activity score. Furthermore, intrahepatic 

CXCL9 and CXCL10 expression was greater in HDV/ 

HBV co-infected patients than in HBV-mono-infected patients. 

To investigate whether HDV specifically induced these T cell-recruiting 

chemokines, we infected Huh-7 cells with recombinant 

HDV and demonstrated the production of CXCL9 and CXCL10 

at both RNA and protein level in a type I IFN-dependent manner. 

In summary, differences in the disease severity of HDV/ 

HBV co-infection and HBV-infection were not attributed to the 

strength of the HDV and HBV-specific T cell response, but to 

HDV’s capacity to specifically induce T-cell recruiting chemokines 

such as CXCL9 and CXCL10. 

Disclosures: 

The following authors have nothing to disclose: Eui-Cheol Shin, Nydiaris Hernandez-Santos, 

Christopher Koh, Mary DeMino, Theo Heller, Barbara Rehermann 

1651 

Deletion of APOBEC3B is Associated With Hepatitis B 

eAg Status in Patients with Chronic HBV Infection 

Alex J. Thompson 1 , Ondrej Podlaha 2 , Zhaoshi Jiang 2 , Xin Guo 2 , 

Luting Zhuo 2 , Dongliang Ge 2 , Anuj Gaggar 2 , Mani Subramanian 

2 , David B. Goldstein 3 , Henry Lik-Yuen Chan 4 , Edward J. 

Gane 5 , Harry L. Janssen 6 ; 1 St. Vincent’s Hospital and the University 

of Melbourne, Melbourne, VIC, Australia; 2 Gilead Sciences, 

Inc., Foster CIty, CA; 3 Institute for Genomic Medicine, Columbia 

University, New York, NY; 4 The Chinese University of Hong Kong, 

Hong Kong, China; 5 Auckland City Hospital, Auckland, New Zealand; 

6 University of Toronto, Toronto General Hospital, Toronto, 

ON, Canada 

Background: The prevalence of HBV infection varies across 

regions, with a higher proportion of HBeAg positive patients 

among Asian populations. APOBEC3 genes play a major 

role in intracellular defense against viral infection including 

HBV. Here we investigated the association between APO- 

BEC3B (A3B) deletions and HBeAg status among Asian and 

Caucasian patients. Methods: Asian (n=70) and Caucasian 

(n= 167) patients with available genomic samples from TDF 

HBV study (GS-US-174-0102/0103) were genotyped using 

Illumina Omini5M+Exome BeadChip platform. A3B deletions 

were inferred from SNP data using the PennCNV software. 

Allele frequency spectrum and linkage equilibrium methods 

were implemented on public human population data (1000 

Genomes Project) to test for signatures of recent positive selec-


tion of A3B deletion. Results: A3B deletions were detected in 

30% of Asian patients and 5% of Caucasian patients. We 

identified a significant association between A3B deletion and 

positive HBeAg status in Asian HBV patients (p = 0.002). 

Patients with this deletion were more likely to be HBeAg positive 

and remained HBeAg positive at an older age (Figure). A 

similar trend was observed among Caucasians patients though 

was not significant, likely due to a low A3B allele deletion frequency 

(p = 0.2). By analyzing the SNP data from the 1000 

Genomes Project, we observed a significant reduction of variation 

around A3B deletion region, suggesting a recent positive 

selection on A3B deletion allele in Asian, Indian, and Caucasian 

populations. Comparing A3B sequences across primates 

revealed a long-term positive selection signature, particularly in 

the human lineage. Conclusion: Structural variation of the APO- 

BEC3B gene is associated with HBeAg status among Asian 

patients. The high frequency of A3B deletion allele in Asian 

populations may be partially explained by positive selection 

forces unrelated to HBV. Further confirmation in independent 

patient cohorts is required. 

Disclosures: 




Abbvie, BMS 

Zhaoshi Jiang - Employment: Gilead Sciences, Inc.; Stock Shareholder: Gilead 

Scieces, Inc. 

Dongliang Ge - Employment: Gilead Sciences, Inc 



David B. Goldstein - Advisory Committees or Review Panels: Sever Adverse 

Events Consortium; Grant/Research Support: Gilead, Astra Zeneca, Biogen, 

Johnson and Johnson, Labcorp, Merck, Fundazione Telethon, NIH, CURE, UCB, 

IL28B and ITPA finds 












The following authors have nothing to disclose: Ondrej Podlaha, Xin Guo, Luting 

Zhuo 

1652 

Effect of a novel synthetic FXR agonist EYP001 on hepatitis 

B virus replication in HepaRG cell line and primary 

human hepatocytes 

Pauline Radreau 2 , Marine Porcherot 1 , Jacky Vonderscher 2 , Vincent 

Lotteau 1 , Patrice André 1 ; 1 CIRI U1111, INSERM, Lyon, France; 

2 EnyoPharma, Lyon, France 

HBV replication is tightly linked to bile acids (BA) metabolism. 

1) BA nuclear receptor FXR binds to two response elements in 

the HBV core promoter region and its activation by ligands 

regulates the HBV core promoter activity 2) the Na-taurocholate 

cotransporting polypeptide (NTCP) responsible of BA 

uptake by hepatocytes was identified as a functional receptor 

for HBV and 3) reciprocally, HBV infection induces changes 

of the expression of several genes involved in the BA synthesis 

pathway in the liver of infected patients. We investigated the 

effect of FXR activity modulation on HBV replication by a novel 

synthetic non-steroidal molecule EYP001 and compare its activity 

to the BA derived 6-ethyl-chenodeoxycholic acid (6-ECDCA) 

and the synthetic FXR agonist GW4064. First, we tested the 

effect of EYP001 1) on the expression of genes under the control 

of FXR in HepaRG cells and in primary human hepatocytes 

(PHH) and 2) on activation of the membrane associated BA 

receptor GpBAR1. Second, we tested the effect of these FXR 

modulators on HBV replication in HepaRG and in PHH, two 

cell culture models that support complete HBV replication cycle. 

Cells were infected with HBV produced by the HepG2.2.15 

line and treated from day 2 to 12 post infection with FXR modulators. 

EYP001 up-regulates the expression of SHP mRNAs 

and down-regulates the expression of the APOA1 mRNA and 

thus behaves as a FXR agonist. In addition treatment for 10 

days also reduces the expression of FXR mRNA, likely by an 

SHP mediated negative feedback. On the opposite, EYP001 

does not activate GpBAR1 and thus appears as a specific 

FXR agonist. Treatment of HBV infected dHepaRG and PHH 

with EYP001 as well as with 6-ECDCA or GW4064 strongly 

inhibited, to similar extent, the secretion of HBV DNA, HBsAg, 

HBeAg and of HBcAg synthesis in a dose dependent manner 

(70 to 80 % inhibition at 1 or 10 microMol) as well as the viral 

pregenomic RNA synthesis, cccDNA copies number and cellular 

total HBV DNA. Cyclosporine A, an NTCP ligand and HBV 

entry inhibitor, did not modify the effect of agonists suggesting 

that the effect did not depend on entry inhibition. Treatment 

consistently reversed the HBV induced modification of FXR and 

related genes expression. In conclusion, this novel FXR agonist 

led to a sustained repression of HBV replication in the HepaRG 

and PHH cell culture systems. This effect was likely mediated by 

the modulation of FXR activation that could perturb the complex 

FXR network of transcription factors, which is highly targeted 

and controlled by HBx. These data stress out the importance to 

exploit drug regulation of metabolism pathways in controlling 

HBV replication. 

Disclosures: 

Pauline Radreau - Employment: ENYO Pharma 

Vincent Lotteau - Consulting: enyopharma; Stock Shareholder: enyopharma 

Patrice André - Stock Shareholder: Enyo Pharma 

The following authors have nothing to disclose: Marine Porcherot, Jacky Vonderscher


1653 

HBcrAg during nucleos(t)ide analog therapy are related 

to intra-hepatic HBV replication and development of 


Masao Honda, Takayoshi Shirasaki, Takeshi Terashima, Kazunori 

Kawaguchi, Mikiko Nakamura, Naoki Oishi, Tetsuro Shimakami, 

Hikari Okada, Kuniaki Arai, Taro Yamashita, Yoshio Sakai, Tatsuya 

Yamashita, Eishiro Mizukoshi, Shuichi Kaneko; Gastroenterology, 

Kanazawa University Graduate School of Medical Science, 


Background & Aim Although nucleos(t)ide analog (NA) therapy 

effectively reduces hepatitis B virus (HBV)-DNA in serum and 

improves liver histology in patients with chronic hepatitis B (CH- 

B), it does not completely reduce the incidence of hepatocellular 

carcinoma (HCC). The objective of this study is to elucidate clinical 

and virological features associated with the development 

of HCC during NA therapy. Material & Methods We enrolled 

109 patients with CH-B who started to receive NA therapy from 

2001 to 2011 at our hospital. All patients received NA therapy 

for more than 2 years, and a liver function test and virological 

markers before treatment and at the end of the follow-up period 

were compared. HBsAg was measured quantitatively using an 

Architect HBsAg-QT assay (Abbot). The amount of HBV-DNA in 

serum was measured with COBAS AmpliPrep-COBAS TaqMan 

HBV Test. HBcrAg was measured by CLEIA using a Lumipulse 

HBcrAg assay (Fujirebio, Inc., Tokyo, Japan). HBV-DNA, HBV- 

RNA and cccDNA in thirteen liver tissue samples taken before 

treatment and 23 liver tissue samples taken during treatment 

were analyzed. Gene expression profiling was performed by 

using an Affymetrix GeneChip. Results During NA therapy for 

6.5 ± 2.8 years, 36 patients (33%) developed HCC. Multivariate 

cox regression analysis showed age (>56 years, HR 

= 3.1) and fibrosis stage (F3–4, HR = 3.4) before treatment 

and the presence of hepatitis core-related antigen (HBcrAg; 

HR = 3.53) during treatment were significantly associated with 

the development of HCC. There was no significant difference 

in the amount of covalently closed circular DNA in HBcrAg(+) 

patients (n = 16) and HBcrAg(-) patients (n = 12); however, 

the amount of HBV-DNA and RNAs (pregenome, preS/S, and 

HBx) were significantly higher in HBcrAg(+) patients than in 

HBcrAg(-) patients, suggesting the presence of active HBV replication 

in HBcrAg(+) liver. Hepatic gene expression profiling 

before and during treatment (n = 13) showed a significant 

improvement of inflammation signaling during treatment; however, 

in HBcrAg(+) liver, viral replication-related signaling such 

as pro-apoptosis and oxidative phosphorylation were up-regulated 

along with cancer-related signaling (n = 11 vs. 12). 

Interestingly, transcription factors that activate HBV promoter 

activity, such as HNF4α and PPARα, were up-regulated in 

HBcrAg(+) liver. Metformin efficiently repressed HBV-RNA and 

HBcrAg levels by repressing the expression of HNF4α and 

PPARα in primary human hepatocytes. Conclusions: Modulating 

HBV transcription factors by metformin in combination with 

NA therapy would potentiate anti-HBV activity and reduce the 

incidence of HCC in HBcrAg (+) patients. 

Disclosures: 







The following authors have nothing to disclose: Masao Honda, Takayoshi Shirasaki, 

Takeshi Terashima, Kazunori Kawaguchi, Mikiko Nakamura, Naoki 

Oishi, Tetsuro Shimakami, Kuniaki Arai, Taro Yamashita, Yoshio Sakai, Tatsuya 

Yamashita, Eishiro Mizukoshi 

1654 

Free episomal and integrated HBV DNA In HBsAg-negative 

patients with intrahepatic cholangiocarcinoma 

Teresa Pollicino 1 , Cristina Musolino 2 , Gianluca Tripodi 2 , Marika 

Lanza 2 , Giuseppina Raffa 2 , Carlo Saitta 2 , Salvatore Benfatto 1 , 

Concetta Beninati 1 , Giuseppe Navarra 3 , Pietro Invernizzi 4 , Domenico 

Alvaro 5 , Giovanni Raimondo 2 ; 1 Pediatric, Gynecological, 

Microbiological and Biomedical Sciences, University of Messina, 

Messina, Italy; 2 Clinical and Experimental Medicine, University 

Hospital of Messina, Messina, Italy; 3 Human Pathology, University 

Hospital of Messina, Messina, Italy; 4 Medicine, Humanitas 

Clinical and Research Center, Milan, Italy; 5 University of Rome 

“Sapienza”, Roma, Italy 

Backgroud: Intrahepatic cholangiocarcinoma (ICC) is a fatal 

primary liver cancer with very poor prognosis. Genome-wide 

studies have made major advances in understanding the molecular 

basis of this disease, although most aspects remain unclear. 

Accumulating evidence indicates that chronic HBV infection is 

associated with an increased risk of ICC development and 

suggests an etiological role of HBV in the development of this 

tumor. Aims of the study were to investigate the prevalence of 

occult HBV infection (OBI) in cases with ICC and to characterize 

the molecular status of HBV in OBI-positive ICC specimens. 

Methods: Frozen paired tumor and non-tumor tissue specimens 

from 40 HBsAg-negative patients with ICC, who underwent 

surgical resection were tested for OBI by 4 different HBV-specific 

nested PCR. To reveal HBV cccDNA, DNA extracts were 

digested with a plasmid-safe ATP-dependent DNase and amplified 

by nested PCR with cccDNA-specific primers. Finally, for 

the detection of HBV DNA integrations the Alu-PCR technique 

was coupled to deep-sequence analysis. Results: HBV genomic 

sequences were detected in tumor and/or non-tumor specimens 

from 28 of the 40 (70%) ICC patients analysed. In particular, 

20/40 (50%) tumors and 13/23 (56.5%) non-tumor tissues 

were HBV DNA positive. HBV cccDNA was detected in tissue 

specimens from 10/28 OBI-positive patients (36%) (both in 

tumor and non-tumor specimens in 3 patients; only in tumor 

tissues in 4 patients; only in non-tumor tissues in 3 patients). 

HBV integrants were detected in 3 of 10 cases examined so 

far, and included portions of the HBx gene sequence (including 

the Basic Core Promoter/Enhancer II) in 2 cases and part of the 

core gene sequence in one case. The analysis of the integration 

sites revealed that the HBx sequences were located 3,374 

nucleotides upstream the sequence encoding the cat eye syndrome 

critical region protein 5 isoform and within the coding 

sequence of the thromboxane A synthase 1, respectively, and 

that the core gene sequence was located within the cystinosin 

isoform 1 precursor coding sequence. Conclusion: Occult HBV 

infection is highly prevalent in patients with ICC. Both free viral 

genomes and integrated HBV DNA can be detected in these 

cases. These results suggest an involvement of HBV in the carcinogenic 

process leading to ICC development even in cases 

with occult infection. 

Disclosures: 

Teresa Pollicino - Speaking and Teaching: Gilead, Roche, Janssen, BMS, MSD, 

Bayer, Abbvie 



The following authors have nothing to disclose: Cristina Musolino, Gianluca 

Tripodi, Marika Lanza, Giuseppina Raffa, Carlo Saitta, Salvatore Benfatto, Concetta 

Beninati, Giuseppe Navarra, Pietro Invernizzi, Domenico Alvaro


1655 

A Novel Entecavir Resistance Mutation RtA186T Causes 

Viral Breakthrough in Chronic Hepatitis B Patients 

Sanae Hayashi 1 , Shuko Murakami 1 , Katsumi Omagari 1 , Takeshi 

Matsui 2 , Etsuko Iio 1 , Masanori Isogawa 1 , Tsunamasa Watanabe 

1 , Yoshiyasu Karino 3 , Yuki Takamatsu 4 , Satoru Kohgo 5 , Kenji 

Maeda 5 , Hiroaki Mitsuya 4,5 , Yasuhito Tanaka 1 ; 1 Department of 

Virology & Liver unit, Nagoya City University Graduate School of 

Medical Sciences, Nagoya, Japan; 2 Center for Gastroenterology, 

Teine Keijinkai Hospital, Sapporo, Japan; 3 Department of Gastroenterology, 

Sapporo Kosei General Hospital, Hokkaido, Japan; 

4 Experimental Retrovirology Section, HIV and AIDS Malignancy 

Branch, National Cancer Institute, National Institutes of Health, 

Bethesda, MD; 5 National Center for Global Health and Medicine, 

Tokyo, Japan 

Background & Aim: Entecavir (ETV) is approved for the first-line 

treatment of chronic hepatitis B virus (HBV) infections due to a 

high genetic barrier. The aim of this study is to characterize 

two novel reverse transcriptase (RT) mutations associated with 

viral breakthrough (VBT) in an ETV-refractory patient. Methods: 

Serum HBV from an ETV refractory patient was sequenced 

before ETV treatment and after VBT. The clones with candidate 

ETV resistance (ETVr) mutations (rtI163T and rtA186T) 

were analyzed for replication efficacy and susceptibility to 

ETV, Adefovir (ADV), Tenofovir Disoproxil Fumarate (TDF), and 

novel nucleoside analogues (NAs) (4’-C-cyano-2-amino-2’-deoxyadenosine 

(CAdA), 4’-C-cyano-2’-deoxyguanosine (CdG)) 

with similar efficacy to ETV in vitro. Moreover, chimeric mice 

with human hepatocytes were inoculated with the patient’s 

serum at VBT, and monitored for viral mutation pattern by 

next-generation sequencing. To confirm the prevalence of the 

novel mutations, 21 ETV-refractory patients were also examined. 

Results: The novel mutations, rtI163V and rtA186T, were 

detected together with LAMr at VBT, but not before ETV treatment. 

RtA186T significantly reduced viral replication efficacy, 

while rtI163V had no impact on it. RtA186T plus LAMr reduced 

susceptibility to ETV as strongly as previously reported ETVr 

mutations rtS202G plus LAMr, resulting in more than 111.1- 

fold resistance to ETV compared with the wild-type clone. In 

contrast, rtI163V plus LAMr resulted in a 20.4-fold resistance 

to ETV, suggesting that rt186T was mainly responsible for the 

VBT. The novel ETVr mutations did not confer cross-resistance 

to ADV, TDF, as well as novel NAs, CAdA and CdG. The viral 

mutation pattern in the chimeric mice indicated dominant proliferation 

of a clone containing rtI163V and rtA186T mutations 

plus LAMr under ETV treatment. In silico docking simulation 

indicated that rtA186T reduced the RT binding ability to ETV. 

In clinical practice, rtA186T, but not rtI163V, was detected 

in another ETV-refractory patient with VBT, suggesting that 

rtA186T could confer ETV resistance independently of rtI163V. 

Conclusion: A novel ETV resistance mutation rtA186T causes 

VBT, and should be closely monitored when chronic hepatitis B 

patients exhibit VBT during prolonged ETV treatment. 

Disclosures: 




Squibb 

The following authors have nothing to disclose: Sanae Hayashi, Shuko Murakami, 

Katsumi Omagari, Takeshi Matsui, Etsuko Iio, Masanori Isogawa, Tsunamasa 

Watanabe, Yuki Takamatsu, Satoru Kohgo, Kenji Maeda, Hiroaki Mitsuya 

1656 

Hepatitis B Virus Surface Protein-induced hPIAS1 Transcription 

Requires TAL1, E47, MYOG, NFI, andMAPK 

Signal Pathways 

Hongyan Wang, Di Wu, Xiaofeng Wang, Guang Chen, Yuanya 

Zhang, Weiming Yan, Meifang Han, Qin Ning; Department and 

institute of Infectious Disease,, Tongji Hospital, Tongji Medical College, 

Huazhong University of Science and Technology, Wuhan, 

China 

The protein inhibitor of activated STAT1 (PIAS1) is ubiquitous 

in mammals and fulfills key functions in cell proliferation and 

inflammation responses. Despite the importance of PIAS1 in 

regulating virus-induced or IFN-stimulated chronic hepatitis, little 

is known about the molecular mechanisms of activation in 

response to hepatic virus infection. Here, we report that the 

hepatitis B virus surface protein (HBs) enhancedhPIAS1 promoter 

activity in luciferase reporter assays. A strong regulatory 

region from −712 to −507 (relative to the transcriptional start 

site) was responsible forhPIAS1 gene transcription in response 

to HBs. TAL1, E47, myogenin (MYOG), and NFI were highly 

expressed and localized to the nucleus in response to HBs. 

Binding of TAL1, E47, MYOG, and NFI to a cis-regulatory 

element in the hPIAS1 promoter was confirmed by electrophoretic 

mobility shift assays and chromatin immunoprecipitation. 

siRNA knockdown of TAL1, E47, MYOG, and NFI 

expression inhibited hPIAS1transcription in response to HBs. 

Increased ERK and p38MAPK phosphorylation was observed 

in HBs-transfected HepG2 cells, and treatment with the ERK 

inhibitor PD098059 or p38MAPK inhibitor SB203580 abolished 

increase in nuclear TAL1, E47, MYOG, and NFI levels 

and hPIAS1 induction. Peripheral blood mononuclear cells from 

patients withhigh HBsAg levels (also withhigh levels of HBV 

DNA ) displayed increased ERK and p38MAPK phosphorylation 

and high levels of TAL1, E47, MYOG, and NFI, compared 

to those with low HBsAg levels or healthy controls. Increased 

hPIAS1 expression was detected in liver biopsies from CHB 

patients with high HBV replication compared to those of undetectable 

for HBV or healthy controls.These findings suggest that 

the HBs protein enhances hPIAS1transcription through activities 

of TAL1, E47, MYOG, and NFI that are dependent on MAPK 

signaling pathways. 

Disclosures: 





The following authors have nothing to disclose: Hongyan Wang, Di Wu, 

Xiaofeng Wang, Guang Chen, Yuanya Zhang, Weiming Yan, Meifang Han 

1657 

The activating receptor NKP46 is essential for the development 

of chronic hepatitis B 

Wanyu Li, Xiuzhu Gao, Ruqi Mei, Jinglan Jin, Junqi Niu; the first 

hospital of jilin university, Changchun, China 

BACKGROUND & AIMS: Our pervious study indicated that 

NKp46 expression regulated NK cell cytolytic function. NKp46 

may moderate NK cell activity during HBV replication suppression 

and HBV-associated liver damage in peripheral blood. 

Here we studied the functions of NKP46 during hepatitis B 

virus (HBV) infection in vivo and in the liver of patients. METH- 

ODS: Intrahepatic NKP46 was investigated in chronic hepatitis 

B patients(n=25) and healthy controls(n=15) by immunohistochemistry.We 

examined the effects of blocking antibodies 

against NKP46(50ug) in immunocompetent mice that express 

HBV from a pAAV/HBV1.2 plasmid(10ug) and are positive


for serum hepatitis B surface antigen (a mouse model of HBV 

infection)(n=10).Goat IgG isotype control antibody was administered 

to control groups.Serum HBsAg titers and HBVDNA 

levels were tested at baseline and post-injection. RESULTS: 

Intrahepatic NKP46 expression is up-regulated in patients with 

CHB compared with healthy control(p=0.032).After injecting 

HBV plasmid,mice that remained HBsAg-positive were used to 

test whether anti-NKP46 mAb treatment affected HBsAg and 

HBVDNA levels during HBV infection.Serum different value 

of HBsAg levels(1508.57±1654.94vs 5597.50±4729.13 

IU/ml;p


entially expressed Genes & miRNAs between patient groups, 

key pathways & biological processes regulated were identified 

by using Volcano plot, miRTarbase & GOElite tool. Differentially 

expressed genes & miRNA along with pathways & 

biological categories regulated, were subjected to Integrome 

analysis to identify disease baseline & stage specific signatures. 

Results: In AVH-B compared to controls,1178 genes,42 

miRNAs were up-regulated & 1348 genes,9 miRNAs were 

down regulated. In CHB-PNALT compared to controls,167 

genes,1 miRNA were-up regulated & 307 genes,11 miRNAs 

were down-regulated. In CHB-RALT compared to controls,82 

genes,4 miRNAs were up-regulated & 58 genes,17 miRNAs 

were down-regulated. In CHB-PNALT compared to AVH-B,500 

genes,9 miRNAs were up regulated & 1307 genes,12 miR- 

NAs were down regulated. In CHB-RALTcompared to AVH-B 

1132 genes,26 miRNAs were up regulated &1694 genes,21 

miRNAs were down regulated. In CHB-RALT compared to CHB- 

PNALT, 183 genes, 3 miRNAs were up regulated & 70 genes, 

3 miRNAs were down regulated.We identified 267 genes & 

32 differentially expressed miRNAs,13 key pathways, biological 

categories & gene families in one or more of the infection 

stagesthat could play major role either in clearance or persistence 

of HBV infection (Fig.1). Conclusion: Integrome analysis 

revealed induction of unique cluster of miRNAs & repression 

of their target genes that differentiates acute & chronic infection 

stage, chronic infection with & without hepatic injury in DCs in 

HBV infection. 

Disclosures: 

The following authors have nothing to disclose: Avishek K. Singh, Robert Geffers, 

Sheetalnath B. Rooge, Aditi Varshney, Madavan Vasudevan, Ankit Bhardwaj, 

Manoj Kumar, Pawan Malhotra, Sunil K. Mukherjee, Raj K. Bhatnagar, Nirupma 

Trehanpati, Shiv K. Sarin 

1660 

Functional restoration of CD56 bright NK cells via IL-15 

and NKG2D correlates with antiviral treatment efficacy 

in chronic hepatitis B patients 

Tao Chen 1 , Aichao Shi 1 , Xiaoping Zhang 1 , Lin Zhu 1 , Zeguang 

Wu 1 , Weiming Yan 1 , Xiaojing Wang 1 , Hong Ma 2 , Jidong Jia 2 , 

Wei Guo 1 , Qin Ning 1 ; 1 Department and Institute of Infectious 

Disease, Tongji Hospital, Tongji Medical College, Huazhong 

University of Science and Technology, Wuhan, China; 2 Beijing 

Friendship Hospital, Beijing, China 

Hepatitis B virus is intrinsically immunogenic, and the immune 

status of the host has a prognostic impact on patients with 

chronic infection and influences the effects of antiviral treatment. 

This current study aimed to investigate the dynamic 

changes of NK cells post antiviral treatment and its potential 

influence on treatment efficacy. This study involved 52 hepatitis 

B e antigen (HBeAg) - positive chronic hepatitis B (CHB) 

patients who received telbivudine (Ldt) for 52 weeks. Blood 

samples were collected at baseline and week 12, 24, 36 and 

48 of treatment, and tested for HBV DNA, HBsAg, HBeAg, ALT, 

AST, and additional immunological markers. Compared with 

baseline, the percentages and absolute number of peripheral 

CD3 - CD56 + NK cells were significantly higher from week 36 

to week 48, especially CD3 - CD56 bright NK cells. The expression 

(percentage and MFI) of activating receptors NKG2D 

and NKP46 was enhanced, while inhibitory receptor NKG2A 

decreased. NKG2D expression was significantly enhanced on 

peripheral NK cells in patients with HBeAg seroconversion, 

particularly in CD3 - CD56 bright NK cell. The serum interleukin 

15 (IL-15) level significantly elevated during Ldt treatment, 

especially in patients with HBeAg seroconversion. Co-culture 

of Ldt with purified peripheral NK cells from treatment naïve 

HBeAg positive CHB patients showed significantly enhanced 

expression of NKG2D and IL-15. These findings indicate that 

antiviral treatment exerted by Ldt in CHB patients may play as 

an “adjuvant” role capable of inducing or restoring NK cell 

function via upregulated NKG2D and IL-15 expression, and 

this in turn correlated with HBeAg seroconversion. 

Disclosures: 

Jidong Jia - Consulting: BMS, GSK, MSD, Novartis, Roche 





The following authors have nothing to disclose: Tao Chen, Aichao Shi, Xiaoping 

Zhang, Lin Zhu, Zeguang Wu, Weiming Yan, Xiaojing Wang, Hong Ma, Wei 

Guo 

1661 

Knockdown of NTCP reduces susceptibility to HBV infection 

in humanized-liver mice 

Tasuku Nakabori, Hayato Hikita, Satoshi Aono, Yoshinobu 

Yokoyama, Kazuhiro Murai, Takuo Yamai, Yugo Kai, Yuki Makino, 

Yasutoshi Nozaki, Yoshinobu Saito, Satoshi Tanaka, Ryotaro 

Sakamori, Tomohide Tatsumi, Tetsuo Takehara; Osaka University 

Graduate School of Medicine, Suita, Japan 

Background and Aim: Hepatitis B Virus (HBV) infects very limited 

species such as humans and chimpanzees, which hamper 

HBV research. Humanized-liver mice have potential to resolve 

these difficulties. Recently, Na+-taurocholate cotransporting 

polypeptide (NTCP) was reported as a candidate of HBV entry 

receptor. However, the effect of its inhibition on HBV infection 

remains unclear. In this study, we clarified it using humanized-liver 

mice. Methods: TK-NOG mice were administrated 

ganciclovir followed by transplantation of human primary 

hepatocytes. Humanized-liver TK-NOG mice (humanized mice) 

were inoculated with HBV obtained from a chronic hepatitis B 

patient. The institutional ethics committee approved the study. 

For in vitro study, primary hepatocytes isolated from humanized 

mice were inoculated with HBV derived from the culture supernatant 

of 2.2.15 cells. Results: Serum HBV-DNA were detected 

in humanized mice from 2 weeks after HBV inoculation. After 

8 to 10 weeks post-inoculation, HBV-DNA levels reached a 

plateau (6-8 log copies/ mL) and almost all of human hepatocytes 

were positive for HBc antigen by immunohistochemistry. 

Administration of siRNA against human NTCP by tail vein successfully 

reduced its expression level in human hepatocytes, 

evidenced by immunohistochemistry and real time RT-PCR. 

After 2 weeks of HBV inoculation following administration of 

NTCP siRNA, the levels of serum HBV-DNA, serum HBs antigen 

and intracellular covalently closed circular DNA (cccDNA) were 

significantly reduced. In sharp contrast, siRNA-mediated NTCP 

knockdown at 12 weeks after HBV inoculation did not affect 

them. Primary hepatocytes from humanized mice were maintained 

in culture over 2 months. After inoculation with HBV for 

24 hours, HBV-DNA, HBs antigen and HBe antigen levels in 

culture medium, gradually increased and kept detectable over 

a month. cccDNA was also detected. Immunohistochemistry


using anti-HBc antibody confirmed their infection with HBV. 

siRNA-mediated knockdown of NTCP efficiently downregulated 

its expression level. Consistent with in vivo data, transfection of 

NTCP siRNA before inoculation with HBV significantly reduced 

HBs antigen, HBe antigen levels in the culture medium, and 

intracellular cccDNA, while its transfection after 20 days of 

HBV inoculation did not affect them. Conclusion: Humanized 

mice and their isolated primary hepatocytes are susceptible to 

HBV infection. Our results suggest that NTCP inhibition would 

be useful for decrease of susceptibility to HBV infection but that 

NTCP inhibition alone might not have an effect on persistent 

infection with HBV. 

Disclosures: 






The following authors have nothing to disclose: Tasuku Nakabori, Satoshi Aono, 

Yoshinobu Yokoyama, Kazuhiro Murai, Takuo Yamai, Yugo Kai, Yuki Makino, 

Yasutoshi Nozaki, Yoshinobu Saito, Satoshi Tanaka, Ryotaro Sakamori, Tomohide 

Tatsumi 

1662 

The Association of vitamin D metabolic pathway 

related genes polymorphisms with virologic response in 

HBeAg-negative patients treated with pegylated interferon: 

Multicenter study 

Kessarin Thanapirom 1 , Sirinporn Suksawatamnuay 1 , Wattana 

Sukeepaisarnjaroen 2 , Tawesak Tanwandee 3 , Satawat Thongsawat 

4 , Teerha Piratvisuth 5 , Rattana Boonsirichan 6 , Chalermrat 

Bunchorntavakul 7 , Chaowalit Pattanasirigool 8 , Bubpha Pornthisarn 

9 , Supot Tantipanichtheerakul 10 , Ekawee Sripariwuth 11 , Woramon 

Jeamsripong 12 , Theeranun Sanpajit 13 , Piyawat Komolmit 1 ; 

1 Medicine, Chulalongkorn University, Bangkok, Thailand; 2 Medicine, 

Khon Kaen University, Khon Kaen, Thailand; 3 Medicine, 

Mahidol University, Bangkok, Thailand; 4 Medicine, Chiang Mai 

University, Chiang Mai, Thailand; 5 Medicine, Prince of Songkla 

University, Songkla, Thailand; 6 Medicine, Vajira Hospital, Bangkok, 

Thailand; 7 Medicine, Rajavithi Hospital, Bangkok, Thailand; 

8 Medicine, Police General hospital, Bangkok, Thailand; 9 Medicine, 

Thammasat University Hospital, Pathum thani, Thailand; 

10 Medicine, Bhumibol Adulyadej Hospital, Bangkok, Thailand; 

11 Medicine, Naresuan University, Phitsanulok, Thailand; 12 Medicine, 

Buddhachinaraj Hospital, Phitsanulok, Thailand; 13 Medicine, 

Phramongkutklao Hospital, Bangkok, Thailand 

Background/aims: In chronic hepatitis B (CHB)-infected 

patients, low serum vitamin D level are associated with high 

level of hepatitis B virus (HBV) replication. From GWAS data, 

the number of single nucleotide polymorphisms (SNPs) within 

the 7-dehydrocholesterol reductase (DHCR7), 1-a-hydroxylase 

(CYP27B1), Cytochrome P450, family 2, subfamily R, polypeptide 

1 (CYP2R1), vitamin D binding protein (GC) and vitamin 

D receptor (VDR) of the vitamin D synthetic pathway is related 

with vitamin D level and function. This study aimed to determine 

the association between the SNPs of vitamin D cascade 

and response to peginterferon (PegIFN) therapy in patients 

with HBeAg-negative CHB infection. Methods: 115 patients 

with HBeAg-negative CHB infection treated for 48 weeks with 

PegIFN-alfa 2a from 13 hospitals were prospectively enrolled. 

Thirteen SNPs across the vitamin D cascade related genes, 

including DHCR7 (rs12785878), CYP27B1 (rs10877012), 

CYP2R1 (rs2060793, rs12794714), GC (rs4588, rs7041, 

rs222020, rs2282679) and VDR (FokI, BsmI, Tru9I, ApaI, 

TaqI) were genotyped. The virologic response was defined 

as HBV-DNA < 2,000 IU/ml. Results: Majority of patients 

(81.7%) had HBV genotype C and 5.3% (n=5) had liver cirrhosis. 

At 24 weeks after therapy, 55.7% (n=54) achieved 

sustained virologic response (SVR) and 6.3% (n=6) cleared 

HBsAg. Seventy-two patients (79.1%) gained end-of-therapy 

virologic response (ETVR). The non-CC allele of FokI (85.3%) 

was significant associated with higher ETVR than CC allele of 

FokI (60.9%)(p


PlasmaBcells(4.1±3.8%Vs12.1±6.45%, p=0.03)&humoral 

immunity were also reduced in Gr A(Fig.B,C).RNA sequencing 

showed decreased expression (>2 fold) of TFH and B cell 

related genes;CD4, BCL6, ICOS, SLAMF1,CXCR5 and BAF- 

F,BATF,CD40, BAFFR in GrA than B(Fig1A).This was validated 

by qRT-PCR showing down-regulation of TFH related genes 

(Fig1D).ROC showed significance of CD4,TFH and B cells 

with cut-off values for prediction of transmission; with highest 

specificity&sensitivity with CD4+ICOS+ & CD4+CXCR5+cells 

as predictors for a transmitting mother(Table).Conclusions:Humoral 

Immunity is essential for viral neutralization.Decreased 

CD4 helper&TFH cells lead to impaired plasma B cell function&facilitate 

HBV transmission to the newborn. Plasma B cell 

can be used as a marker to predict HBV Transmission. 

beclin-1 (6 fold), SIRT-1 (4 fold), c-myc (4 fold), and PTEN with 

HA treatment (100 μg/ml). The expression of p-akt, p-erk-1/2 

were also inhibited while the expression of the caspase-3 and 

β-catenin were found to be up-regulated. The autophagosome 

formation was inhibited significantly (p


1666 

Presence of naïve-like CD8+ T cells specific for subdominant 

hepatitis B virus epitopes in chronically infected 

patients 

Anita Schuch 1,2 , Muthamia Kiraithe 1,2 , Julia Lang 1 , Christoph Neumann-Haefelin 

1 , Robert Thimme 1 ; 1 Department of Medicine II, University 

Hospital Freiburg, Freiburg, Germany; 2 Faculty of Biology, 

University of Freiburg, Freiburg, Germany 

In chronically HBV-infected patients, virus-specific CD8+ T 

cells are rarely detectable ex vivo by conventional tetramer 

stainings. We aimed to analyze whether his lack of HBV-specific 

CD8+ T-cell responses may be explained by insufficient 

priming or terminal deletion. To phenotypically characterize 

HBV-specific CD8+ T cells that are not detectable by conventional 

methods we used a combination of tetramer-associated 

magnetic bead enrichment and multiparametric flow cytometry 

and additionally performed viral sequence analyses. We analyzed 

CD8+ T-cell populations specific for 4 well-described 

HLA-A*02:01 restricted HBV epitopes (Core 18-27 

, Env 183-191 

, 

Env 335-343 

and Pol 455-463 

) in a heterogeneous cohort of 34 

chronically HBV-infected patients. We were able to detect 108 

of 136 (79%) HBV-specific CD8+ T-cell populations. Of note, 

each patient had at least one detectable CD8+ T-cell population, 

suggesting that these cells are not completely deleted 

in chronic infection. Enriched CD8+ T cells specific for the 

immunodominant Core 18-27 

epitope predominantly showed an 

antigen-experienced effector-memory phenotype (CD45RA - , 

CCR7 - , CD27 + , CD11a + ) which was also detectable in patients 

with resolved HBV infection. In contrast, CD8+ T cells specific 

for Env 183-191 

, Env 335-343 

and Pol 455-463 

often displayed a naïvelike 

phenotype (15%, 44% and 19% respectively) defined by 

high expression of CD45RA, CCR7 and CD27. Absence of 

CD11a and CD95 expression additionally indicated a naïve 

status of these enriched HBV-specific CD8+ T cells. Importantly, 

the presence of wild type sequences in patients with naïve-like 

HBV-specific CD8+ T cells indicated that the presence of naïvelike 

cells is not due to infection with a variant HBV containing 

mismatched epitopes. It is also important to note that the presence 

of HBV-specific CD8+ T cells with naïve-like phenotype 

did not correlate with any clinical parameter such as viral load, 

transaminase levels or HBeAg status, and was independent 

of antiviral treatment. Taken together, our results indicate that 

HBV-specific CD8+ T cells are not completely deleted in chronically 

infected patients, but are rather maintained at a very low 

frequency. Furthermore, a substantial fraction of CD8+ T cells 

specific for subdominant HBV epitopes display a naïve-like 

phenotype despite ongoing viral replication. This suggests that 

insufficient priming and/or antigen ignorance of HBV-specific 

CD8+ T cells may also contribute to CD8+ T-cell failure in 

chronic HBV infection and that therapeutic vaccination may be 

a feasible approach in this patient cohort. 

Disclosures: 

Christoph Neumann-Haefelin - Advisory Committees or Review Panels: AbbVie; 

Speaking and Teaching: AbbVie, Gilead, Bristol-Myers Squibb 

The following authors have nothing to disclose: Anita Schuch, Muthamia Kiraithe, 

Julia Lang, Robert Thimme 

1667 

Precore and Basal Core Promoter mutations in HBeAg 

negative chronic hepatitis B virus infection in Cape 

Town, South Africa 

Mark W. Sonderup 1 , Heidi Smuts 2 , Ruud A. Roos 3,1 , Helen 

Wainwright 4 , Neliswa A. Gogela 1 , Mashiko Setshedi 1 , Henry N. 

Hairwadzi 1 , C. W. Spearman 1 ; 1 Department of Medicine and 

Division of Hepatology, University of Cape Town, Cape Town, 

South Africa; 2 Division of Medical Virology, University of Cape 

Town and NHLS, Cape Town, South Africa; 3 Vrije Universiteit, 

Amsterdam, Netherlands; 4 Department of Anatomical Pathology, 

University of Cape Town, Cape Town, South Africa 

Chronic HBeAg negative HBV represents the immune control 

or escape phase of infection. Precore (PC) and basal core 

promoter (BCP) mutations are associated with progressive liver 

disease or HCC although this is poorly studied in sub-Saharan 

Africa. Our aim was to characterize mutations in HBeAg negative 

patients. Methods: Prospectively, over 2 years, HBeAg negative 

patients were analysed by amplifying viral PC and BCP 

regions using nested primers. Amplicons were direct Sanger 

sequenced in the forward & reverse directions and compared 

to reference sequences. Relevant demographic, clinical, virological 

and histological data were captured. Results: 124 

patients, median age 35.1 yrs (IQR 29-43.2), 60.5% (n=75) 

male, 3.2% (n=4) HIV co-infected, were analysed. Ethnically, 

52% were mixed ancestry, 28% Black African, 16% White 

and 4% Asian. Genotype distribution - A 51%, B 2.3%, C 

1.1%, D 42% and E 3.6%. Mutations were detected in 98.4% 

(n=122); 89% (n=109) with PC; 53.3% (n=65) BCP and 46% 

(n=56) both. Mutation frequencies are listed. Median ALT (U/L) 

was significantly higher when both PC/BCP than PC only mutations 

were present, [44 (IQR 41-47); 27 (19-52)] p=0.0007, 

but not with BCP only mutations [32 IQR (22-65)]; p=0.069. 

With dual A1762T G1764A BCP mutations, median ALT and 

HBV DNA (log 10 

IU/ml) was significantly higher compared to 

the T1753C only BCP mutation, [52 (IQR 25-134); 24 (IQR21- 

49)]; p=0.019 and [5.2 (IQR 4-7.6); 3.8 (3-5.3)]; p=0.02. 

In those biopsied (n=31), median Ishak necro-inflammatory 

stage and fibrosis grade scores were 5 (IQR 3-7) & 2 (IQR 1-3) 

respectively with no differences observed in PC, BCP or PC/ 

BCP groups. Six patients, 4.8%, developed HCC with 5 (83%) 

having both PC/BCP mutations, 3 (50%) the dual A1762T 

G1764A BCP mutation and 4 (66%) the G1896A PC mutation. 

Conclusion: In HBeAg negative, mostly A or D genotype CHB 

patients, PC were present more frequently than BCP mutations; 

almost half having both. Dual PC/BCP predicted for higher ALT 

and HBV DNA; a similar trend observed for the dual A1762T 

G1764A BCP mutation. Mutation analysis may have a role in 

guiding the need for antiviral therapy.


Disclosures: 

The following authors have nothing to disclose: Mark W. Sonderup, Heidi Smuts, 

Ruud A. Roos, Helen Wainwright, Neliswa A. Gogela, Mashiko Setshedi, Henry 

N. Hairwadzi, C. W. Spearman 

1668 

Differential Serum Cytokine Profiles in Patients with 

Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), and 

Non-Viral Non-Autoimmune Liver Disease, with or without 

Hepatocellular Carcinoma (HCC) 

Vincent L. Chen 2 , Philip Vutien 3,5 , Biao Li 4 , Ondrej Podlaha 4 , 

Ellen T. Chang 5 , Zhaoshi Jiang 4 , Dongliang Ge 4 , Anuj Gaggar 4 , 



of Medicine, Stanford University Medical Center, Stanford, 

CA; 3 Rush University Medical Center, Chicago, IL; 4 Gilead Sciences, 

Inc., Foster City, CA; 5 Department of Health Research and 

Policy (Epidemiology), Stanford University School of Medicine, 

Stanford, CA 

Aims: HBV and HCV are the most common causes of HCC. 

Differences in immune responses to HBV, HCV, non-viral disease, 

and HCC remain incompletely defined. Our goal was 

to compare the cytokine profiles of patients with HBV, HCV, or 

non-viral liver disease, with or without HCC. Methods: Serum 

samples were obtained from 381 patients seen at a US medical 

center from 2000-2007: 78 patients with HCC (33 HBV, 

45 HCV) and 303 without HCC (119 HBV, 131 HCV, 53 

non-viral). Microplex analysis (Luminex 200 IS) was used to 

measure serum levels of 51 common cytokines. Random forest 

machine learning was used to generate receiver operator 

characteristic (ROC) curves and determine individual cytokine 

importance, using Z scores of mean fluorescence intensity for 

individual cytokines. Results: Among non-HCC patients, 60% 

were male with mean age 52 years. Most HBV patients were 

Asian (89%), while only 18% of HCV and 4% of non-viral 

patients were Asian. Among HCC patients, 85% were male 

with mean age 62 and similar ethnic distribution. Cytokine concentrations 

distinguished non-HCC HBV and HCV patients, with 

area under the ROC curve (AUC) of 0.91 (Fig. 1A). The cytokines 

with greatest predictive power were soluble FasL, M-CSF, 

TNF-β, and IP-10. Cytokine patterns also differed between non- 

HCC HBV and non-viral patients (Fig. 1A; AUC 0.84), and the 

most important cytokines were leptin, resistin, IL-8, and M-CSF. 

Cytokine profiles were less distinct between HCV vs. non-viral 

liver disease (AUC 0.72). Of note, the cytokine pattern of HCC 

patients was different from that of non-HCC patients in HBV 

(Fig. 1B; AUC 0.77) but not HCV cohort (Fig. 1B; AUC 0.48). 

Finally, cytokine profiles differed between non-HCC patients 

with alanine aminotransferase ≥ 2x vs. < 2x upper limit of normal 

(AUC 0.75) but not between Asian and non-Asian patients 

(AUC 0.55 for HBV and 0.51 for HCV patients). Conclusions: 

Serum cytokine profile distinguishes non-HCC patients with 

HBV from those with HCV or non-viral disease, and between 

HBV patients with and without HCC. Future work will expand 

the cohort, perform additional subgroup analysis, and identify 

cytokine profiles that correlate with disease progression. 

Disclosures: 

Biao Li - Employment: Gilead Sciences, Inc. 

Ellen T. Chang - Employment: Exponent, Inc. (a for-profit science and engineering 

consulting company that works on behalf of numerous health care clients, among 

others) 

Zhaoshi Jiang - Employment: Gilead Sciences, Inc.; Stock Shareholder: Gilead 

Scieces, Inc. 







The following authors have nothing to disclose: Vincent L. Chen, Philip Vutien, 

Ondrej Podlaha 

1669 

Possible involvement of hepatic steatosis in the elevation 

of aminotransferases in hepatitis B virus-infected 

patients who are HBeAg-negative and have low DNA 

levels 

Hirayuki Enomoto, Nobuhiro Aizawa, Yoshiyuki Sakai, Naoto 

Ikeda, Kazunori Yoh, Ryo Takata, Chikage Nakano, Kunihiro Hasegawa, 

Kyohei Kishino, Yoshihiro Shimono, Akio Ishii, Tomoyuki 

Takashima, Takashi Nishimura, Hiroki Nishikawa, Yoshinori 

Iwata, Hiroko Iijima, Shuhei Nishiguchi; Division of Hepatobiliary 

and Pancreatic Medicine, Department of Internal Medicine, Hyogo 

College of Medicine, Nishinomiya, Japan 

Background: The clinical significance of hepatic steatosis in 

hepatitis B virus (HBV)-infected patients who are HBeAg-negative 

and have low HBV-DNA levels (referred to as asymptomatic 

carriers) has not been sufficiently clarified. Methods: 

Among a total of 112 treatment-naïve HBV-infected patients 

with a liver biopsy, we assessed the histological and laboratory 

findings to evaluate whether hepatic steatosis and its 

related metabolic disorders were associated with an elevation 

in ALT levels in HBeAg-negative patients (N=70). The histological 

degrees of hepatic steatosis were classified as Grade 0 

(33%-66%), and Grade 

3 (>66%) as described by Kleiner et al. Clinical variables with 

suggested associations with hepatic steatosis, such as the levels 

of homeostatic model assessment insulin resistance (HOMA-IR) 

and serum ferritin were measured. We also calculated the 

NAFIC score, which is a simple scoring system determined by 

the levels of three variables (serum ferritin concentration, fasting 

insulin, and type IV collagen 7S) that has been proposed 

for predicting nonalcoholic steatohepatitis (NASH) in Japanese 

patients. The study conformed to the ethical guidelines 

of the Declaration of Helsinki, and written informed consent 

was obtained from all patients on admission. This study was 

approved by the ethics committee of the institutional review 

board at our institution. Results: In HBeAg-negative patients 

with high HBV-DNA levels (HBV-DNA ≥ 2000 IU/ml), the elevation 

of the ALT levels was related to the HBV-DNA levels 

independent of the histological severity of hepatic steatosis


and the levels of hepatic steatosis-related metabolic variables. 

However, in HBeAg-negative patients with low DNA levels 

(HBV-DNA < 2000 IU/ml), the ratio of patients with moderate 

to severe hepatic steatosis (Grades 2 or 3) and the levels of 

hepatic steatosis-related variables were significantly higher in 

patients with elevated ALT levels than those without (steatosis: 

53.8% vs. 5.0%, p


untreated cells. Finally, the HBV enhancer-1 activity was examined 

by luciferase assay using luciferase vector inserted HBV 

enhancer-1 region. GGA treatment resulted in approimately 

50% reduction of HBV enhancer-1 activity in Huh7 cells compared 

to untreated cells. (Conclusion) GGA reduces HBV 

related protein and mRNA in Huh7 hepatoma cells by suppressing 

HBV enhancer-1 activity. GGA may increase anti-HBV 

effect in combination with other therapies. 

Disclosures: 

The following authors have nothing to disclose: Masafumi Haraguchi, Satoshi 

Miuma, Yuko Akazawa, Hidetaka Shibata, Takuya Honda, Hisamitsu Miyaaki, 

Naota Taura, Tatsuki Ichikawa, Kazuhiko Nakao 

1672 

Dynamic Analysis Of Cellular Factors Modulated By 

Hepatitis B Virus Protein Expression And Replication 

Kenichi Morikawa 1 , Tomoe Shimazaki 1 , Takaaki Izumi 1 , Machiko 

Umemura 1 , Masato Nakai 1 , Goki Suda 1 , Darius Moradpour 2 , 

Naoya Sakamoto 1 ; 1 Department of Gastroenterology and Hapatology, 

Hokkaido University, Sapporo, Japan; 2 University of Lausanne, 

Lausanne, Switzerland 

Background: Many viruses manipulate host factors for their 

purposes. The hepatitis B virus (HBV) is believed to regulate 

host factors for viral replication, persistence and pathogenesis. 

However, the mechanisms by which HBV establishes and 

maintains chronic infection are poorly understood. The aim of 

this study was to perform a dynamic analysis of cellular factors 

modulated by HBV for a better understanding of the life cycle 

and pathogenesis of HBV. Methods: Huh7 cells inducibly replicating 

HBV were analyzed by stable isotopic labeling using 

amino acids in cell culture (SILAC) coupled with mass spectrometry. 

In these cells, the core and the polymerase are inducibly 

expressed upon tetracycline withdrawal from the culture 

medium while pre-S1, pre-S2, and S as well as X are constitutively 

expressed from endogenous HBV promoters. Metabolites 

were analyzed by mass spectrometry in the same cell lines. 

Oxidative stress was assessed by the GSH/GSSG ratio. Mitochondrial 

superoxide was measured by MitoSOX using flow 

cytometry. Morphological changes of the endoplasmic reticulum 

and mitochondria, as well as autophagosome induction 

were visualized by negative stain transmission electron microscopy 

(TEM). Results: SILAC coupled with mass spectrometry 

identified > 3800 proteins, and mass spectrometry measured 

112 metabolites. We observed an increase in the abundance 

of mitochondrial proteins and a decrease of ribosomal proteins 

upon replication of HBV. There was no significant difference 

in mitochondrial morphology between cells replicating 

vs. not replicating HBV. More cells containing lysosomes and 

autophagosomes were observed in the presence of replicating 

HBV. HBV replication was associated with the production of 

mitochondrial superoxide. However, only a slight increase of 

oxidative stress was noted. Conclusions: The observed changes 

suggest a slow down or stop of cell cycle and proliferation for 

viral fitness. The defense mechanism against oxidative stress 

induced by HBV might be maintained by autophagy. Analyses 

of cellular dynamics should yield new insights into the HBV life 

cycle and the pathogenesis of hepatitis B and may reveal novel 

angles for therapeutic intervention. 

Disclosures: 



The following authors have nothing to disclose: Kenichi Morikawa, Tomoe Shimazaki, 

Takaaki Izumi, Machiko Umemura, Masato Nakai, Goki Suda, Darius 

Moradpour 

1673 

IFN-λ3 induction by nucleotide, not nucleoside, analogs 

and its clinical significance in HBV patients 

Kazumoto Murata 1 , Akihiro Matsumoto 2 , Taisuke Inoue 3 , Minoru 

Sakamoto 3 , Masaya Sugiyama 1 , Nobuyuki Enomoto 3 , Eiji 

Tanaka 2 , Masashi Mizokami 1 ; 1 The Research Center for Hepatitis 

and Immunology, National Center for Global Health and Medicine, 

Ichikawa, Japan; 2 Department of Medicine, Shinshu University 

School of Medicine, Matsumoto, Japan; 3 First Department of 

Medicine, University of Yamanashi, Chuo, Japan 

BACKGROUND & AIM: Clinical significances of IL-28B polymorphisms 

in patients with hepatitis B virus (HBV) infection are 

controversial while its pivotal roles were established in chronic 

hepatitis C. Therefore, we sought to clarify the physiological 

roles of its protein, IFN-λ3 levels in HBV patients. METHODS & 

RESULTS: We measured serum IFN-λ3 levels together with IFNλ1 

or IFN-λ2 in asymptomatic carriers (n=83), chronic hepatitis 

(n=113), and liver cirrhosis (n=58). No patients were treated 

with IFN when serum was taken. IFN-λ3 was measured by 

our newly developed chemiluminescence enzyme immunoassay 

(CLEIA) kit, which demonstrated little or no cross reactivity 

between IFN-λ2 and IFN-λ3. IFN-λ1 or IFN-λ2 was measured 

by commercial ELISA kit. No significant differences in serum 

IFN-λ3 levels were observed in disease progression, IL-28B 

polymorphisms or HBeAg positivity. Unexpectedly, however, 

we found higher serum IFN-λ3 levels in patients treated with 

adefovir pivoxil (ADV) or tenofovir disoproxil fumarate (TDF), 

compared with lamivudine (LAM) or entecavir (ETV) (35.4 ± 

27.6 pg/ml vs 3.0 ± 4.9 pg/ml, p < 0.0001) although no differences 

in their clinical background were observed. IFN-λ1 or 

IFN-λ2 did not reveal these associations. Serial measurements 

of serum IFN-λ3 in each patient demonstrated its elevation after 

initiation of ADV or TDF and decline after cessation of it, suggesting 

ADV or TDF itself induced IFN-λ3. These were further 

confirmed by in vitro experiments with colon cancer cell lines 

that ADV or TDF, not LAM or ETV, induced IFN-λ3 in a dose 

dependent manner with additive effects of IFN. In addition, 

immunohistochemistry of IFN-λ3 showed clear staining in the 

cytoplasm. No nucleos(t)ide analogs (NUC) induced IFN-λ3 in 

other cell lines originated from blood, liver, lung, and skin. In 

another setting, 83 HBV patients were prospectively switched 

to PEG-IFN treatment for 48 weeks after cessation of NUC 

(sequential therapy). Low HBsAg (< 3.0 Log IU/ml) and high 

serum IFN-λ3 (>19.0 pg/ml) before PEG-IFN were identified 

as significant factors associated with HBsAg reduction at the 

end of PEG-IFN by logistic regression analysis. CONCLUSIONS: 

This is a first report that nucleotide analogs (ADV or TDF), not 

nucleoside analogs (LAM or ETV), have an additional effect 

to induce IFN-λ3 as well as HBV polymerase inhibition. It is 

possible that IFN-λ3 induced in the gastrointestinal tracts by 

orally administered ADV or TDF reaches to the liver via portal 

tract and has immuno-modulatory effects against HBV. These 

results strongly indicate that prospective studies with additional 

PEG-IFN on ADV or TDF after achievement of 2 factors above 

would be warranted. 

Disclosures: 

The following authors have nothing to disclose: Kazumoto Murata, Akihiro Matsumoto, 

Taisuke Inoue, Minoru Sakamoto, Masaya Sugiyama, Nobuyuki Enomoto, 

Eiji Tanaka, Masashi Mizokami


1674 

Long-term follow-up study of hepatitis delta virus quasispecies 

complexity 

Maria Homs 1,2 , Maria Buti 1,3 , Alicia Ruiz 4 , Pilar Reimundo 4 , Josep 

Gregori 5 , Maria Blasi 1,2 , Rosario Casillas 5 , David Tabernero 1,2 , 

Marta Vila 5 , Leonardo Nieto 2 , Josep Quer 1,5 , Mar Riveiro-Barciela 

3 , Rafael Esteban 1,3 , Francisco Rodriguez-Frias 1,2 ; 1 Centro 

de Investigacion Biomedica en Red de Enfermedades Hepaticas y 

Digestivas, CIBERehd, Barcelona, Spain; 2 Microbiology, Hospital 

Vall d’Hebron, Barcelona, Spain; 3 Hepatology, Hospital Vall d’Hebron, 

Barcelona, Spain; 4 Biochemistry, Hospital Vall d’Hebron, 

Barcelona, Spain; 5 Liver Diseases, Research Institute of Vall d’Hebron, 


Background The hepatitis delta virus (HDV) genome encodes 

two antigens essential for viral cycle, small and large (S- and 

L-HDAg). Genomes with a stop codon in position 196 encode 

S-HDAg, and with a tryptophan encode L-HDAg. Immune 

responses likely play a key role in controlling HDV infection; 

but changes occurring in the quasispecies (QS) complexity and 

in the percentage of genomes encoding S-HDAg and L-HDAg 

in the circulating HDV QS are unknown. Aim Evaluate changes 

in the complexity of HDV QS and in the percentage of genomes 

encoding S-HDAg and L-HDAg during long-term chronic HDV 

infection. Patient and methods Twelve consecutive serum samples 

from a patient with chronic HDV followed for 12 years 

were included. The patient was HDV genotype 1, HBeAg-negative, 

and HBV-DNA levels were


1676 

Noninvasive markers of liver fibrosis remain stable in 

the majority of hepatitis B virus and human immunodeficiency 

virus co-infected patients undergoing tenofovir-containing 

antiretroviral therapy 

Anders Boyd 1 , Karine Lacombe 1,2 , Pierre-Marie Girard 1,2 , Caroline 

Lascoux-combe 3 , Patrick Miailhes 4 , Hayette Rougier 2 , Lawrence 

Serfaty 2 ; 1 Inserm UMR_S1136, Paris, France; 2 Hôpital 

Saint-Antoine, Paris, France; 3 Hôpital Saint-Louis, Paris, France; 

4 Hôpital Croix-Rousse, Lyon, France 

Long-term tenofovir (TDF) use has been associated with significant 

regression of liver fibrosis during hepatitis B virus (HBV) 

monoinfection. However, there are few data in TDF-treated 

patients infected with both HBV and the human immunodeficiency 

virus (HIV), in whom HIV-associated immunosuppression 

could affect liver repair. In this prospective study, 168 HIV-HBV 

co-infected patients enrolled in the French HIV-HBV cohort were 

included if they initiated TDF-containing antiretroviral therapy 

and had liver fibrosis measurements at baseline and ≥1 during 

treatment. Patients with hepatitis C of D virus were not included. 

Fibrosis was assessed at baseline and every 6-12 months and 

≥F3 fibrosis determined from two noninvasive measures: biochemical 

score (≥0.59, FibroTest®) and/or transient elastography 

(≥7.6kPa, FibroScan). Determinants for fibrosis regression 

or progression were evaluated using Cox proportional hazards 

regression. Patients were predominately male (86.3%) with 

a median age of 41 years (IQR=36-48). At baseline, HBV- 

DNA was detectable in 134 (80.2%) patients with a median 

HBV-DNA viral load of 5.02 IU/mL (IQR=2.95-7.15) and 104 

(61.9%) had HBeAg-positive serology. After a median follow-up 

of 59.3 months (IQR=36.1-92.7), 140 (83.3%) patients 

were able to maintain or achieve undetectable HBV-DNA viral 

loads. Among patients with F3-F4 baseline liver fibrosis, 24/53 

(45.3%) remained at the same level of fibrosis during treatment 

and 29/53 (54.7%) regressed to F0-F1-F2 fibrosis after 

a median 16.9 months (IQR=8.6-32.8). Of the latter group, 

10/29 (34.5%) maintained F0-F1-F2 fibrosis until the end of 

follow-up. Liver fibrosis regression was significantly associated 

with CD4+ cell count ≥500/mm 3 (HR=2.09, 95%CI=1.13- 

3.88) in multivariable analysis. Among patients with F0-F1-F2 

baseline liver fibrosis, 70/115 (60.9%) remained at the same 

level of fibrosis and 45/115 (39.1%) progressed to F3-F4 

fibrosis after a median 29.0 months (IQR=18.0-47.4). Of the 

latter group, 28/45 (59.6%) maintained F3-F4 fibrosis. Liver 

fibrosis progression was significantly associated with higher 

age (HR/year=1.10, 95%CI=1.06-1.14), alanine aminotransferase 

>2X ULN (HR=2.10-9.28), and tended to be associated 

with an AIDS-defining event (HR=1.79, 0.98-3.27) in 

multivariable analysis. In conclusion, liver fibrosis levels are 

stable for the majority of HIV-HBV co-infected patients undergoing 

TDF-containing ART, despite extensive control of HBV 

replication. Immunosuppression plays a strong role in reducing 

fibrosis levels, supporting the need for earlier ART-initiation in 

this patient population. 

Disclosures: 

Lawrence Serfaty - Advisory Committees or Review Panels: MSD, Janssen, Roche, 

Gilead, BMS, Abbvie; Speaking and Teaching: Aptalis 

The following authors have nothing to disclose: Anders Boyd, Karine Lacombe, 

Pierre-Marie Girard, Caroline Lascoux-combe, Patrick Miailhes, Hayette Rougier 

1677 

Factors associated with decreases in estimated glomular 

filtration rates for patients co-infected with hepatitis B 

virus and human immunodeficiency virus undergoing 

tenofovir-containing antiretroviral therapy 

Anders Boyd 1 , Karine Lacombe 2,1 , Patrick Miailhes 3 , Caroline 

Lascoux-combe 4 , Hayette Rougier 2 , Julie Bottero 2,1 , Pierre-Marie 

Girard 2,1 ; 1 Inserm UMR_S1136, Paris, France; 2 Hôpital Saint-Antoine, 

Paris, France; 3 Hospices Civils de Lyon, Lyon, France; 4 Hôpital 

Saint-Louis, Paris, France 

Renal toxicity remains one of the more common tolerance 

issues encountered during prolonged treatment with tenofovir 

(TDF). Although determinants for renal impairment during TDF 

have been well established among patients infected with hepatitis 

B virus (HBV) or human immunodeficiency virus (HIV), little 

is known how these specific factors influence renal function in 

HIV-HBV co-infected patients. In this prospective study, 175 

HIV-HBV co-infected patients enrolled in the French HIV-HBV 

cohort were included if they initiated TDF-containing antiretroviral 

therapy (ART) and had creatinine levels available at 

baseline and ≥1 during treatment. Patients with hepatitis C 

or D virus were not included. Estimated glomular filtration 

rates (eGRF) were calculated using the CKD-EPI equation 

from creatinine values obtained at baseline and every 6-12 

months. Mixed-effect linear regression models were used to 

evaluate differences in eGRF changes from baseline (ΔeGFR) 

between certain risk-factors. Determinants towards impaired 

renal function (90 (p=0.002), males (p=0.04), an AIDS-defining illness 

(p=0.03), shorter duration of ART at baseline (p=0.01), concomitant 

therapy with a protease inhibitor (p=0.005), and 

baseline liver cirrhosis (p=0.03). Among patients with normal 

baseline renal function, 51/114 (44.7%) maintained eGFR 

levels >90 mL/min/1.73m 2 , while 63/114 (55.3%) developed 

either mild (60-89 mL/min/1.73m 2 , n=59) or moderate 

(30-59 mL/min/1.73m 2 , n=4) renal impairment after 15.0 

months (IQR=7.1-35.8). In multivariable analysis, higher age 

(HR/year=1.05, 95%CI=1.01-1.10) and AIDS-defining illness 

(HR=1.68, 95%CI=1.05-2.69) were associated with developing 

renal impairment, whereas undetectable HBV-DNA was 

significantly protective (HR=0.41, 95%CI=0.23-0.76). In conclusion, 

liver fibrosis and AIDS-defining illnesses negatively 

and independently affect renal function in HIV-HBV co-infected 

patients undergoing long-term TDF. Controlled HBV-infection 

prevents further progression towards renal impairment, stressing 

the need for effective therapy in this population. 

Disclosures: 

The following authors have nothing to disclose: Anders Boyd, Karine Lacombe, 

Patrick Miailhes, Caroline Lascoux-combe, Hayette Rougier, Julie Bottero, 

Pierre-Marie Girard


1678 

No Detectable Resistance to Tenofovir Disoproxil Fumarate 

(TDF) when Given Alone or in Combination with 

Emtricitabine (FTC) in Chronic Hepatitis B (CHB) Patients 

with Documented Resistance to Lamivudine (LAM): Final 

5 Year Results 

Thomas Berg 2 , Edward J. Gane 3 , Maciej S. Jablkowski 4 , Petr 

Urbanek 5 , Amoreena C. Corsa 1 , Yang Liu 1 , Kyungpil Kim 1 , John 

F. Flaherty 1 , Cihan Yurdaydin 6 , Scott Fung 7 , Kathryn M. Kitrinos 1 ; 

1 Gilead Sciences, Foster City, CA; 2 University Hospital Leipzig, 

Leipzig, Germany; 3 New Zealand Liver Transplant Unit, Auckland, 

New Zealand; 4 Medical University of Lodz, Lodz, Poland; 5 Central 

Military Hospital Prague, Prague, Czech Republic; 6 Ankara University 

Medical School, Ankara, Turkey; 7 Toronto General Hospital, 


Aim: To compare amino acid changes within HBV polymerase/reverse 

transcriptase (pol/RT) after up to 5 years (240 

weeks) of treatment with TDF or the combination of FTC and 

TDF. Methods: Patients enrolled in the study were receiving 

LAM, harbored LAM resistance mutations in pol/RT (rtM204V/ 

I±rtL180M) as determined by INNO-LiPA Multi-DR v2/v3, and 

were randomized 1:1 to receive TDF or FTC/TDF for up to 

240 weeks. A subset of enrolled patients received prior CHB 

treatment in addition to LAM: 13 received entecavir (ETV) and 

61 received adefovir (ADV). In addition, a subset of enrolled 

patients harbored ETV resistance mutations at baseline (TDF = 

14, FTC/TDF = 11). Virologic breakthrough (VB) was defined 

as confirmed HBV DNA >1 log 10 

increase from nadir or HBV 

DNA ≥69 IU/mL after


Disclosures: 








The following authors have nothing to disclose: Yuji Ishida, Chihiro Yamasaki, 

Ami Yanagi, Yasumi Yoshizane, Kazuyuki Fujikawa, Koichi Watashi, Hiromi 

Abe, Takaji Wakita, Nelson Hayes, Chise Tateno 

1680 

Modeling Hepatits B Virus infection in stem cell derived 

hepatocytes reveals role of perturbations in NTCP genotype 

and expression in HBV permissiveness 

Angela Frankel, Robert E. Schwartz; Weill Cornell Medical College, 

New York, NY 

Worldwide, hepatitis B virus (HBV) infection is the most common 

viral hepatitis having infected over two billion people and 

chronically infecting more than 400 million, putting them at 

increased risk to develop cirrhosis and hepatocellular carcinoma. 

HBV research has been hampered by the virus’s narrow 

host range and cellular tropism for hepatocytes, which has led 

to a paucity of robust and reliable infectious systems for HBV 

study. Current model systems to study HBV include hepatoma 

cell lines which do not faithfully recapitulate adult hepatocyte 

phenotype or function as they have undergone a variety of 

genetic and metabolic changes. As a consequence, major components 

of the viral entry process and viral life cycle - including 

the establishment and persistence of a nuclear cccDNA pool 

- and many aspects of virus-host interactions have been poorly 

understood. We have recently shown that micropatterned 

co-cultures of primary human hepatocytes with stromal cells 

and inducible pluripotent stem cells differentiated into hepatocyte-like 

cells (iHeps) are permissive to HBV infection. iHeps 

become permissive only during late stages of differentiation, 

following the activation of the HBV transcription machinery, 

and the appearance of the recently discovered HBV receptor 

sodium-taurocholate co-transporter polypeptide (NTCP). Mutations 

of NTCP have been shown to severely impair bile acid 

uptake but the impact that particular variants play in HBV infection 

is largely unknown. In particular the NTCP Ser267Phe variant 

has recently been shown to be associated with resistance 

to the development of chronic HBV in a recently completed 

genome-wide association study. We hypothesized that mutations 

in NTCP may be directly responsible for this association. 

A gene editing approach using CrispR (and guide RNA’s targeting 

the NTCP locus) along with homology directed repair 

enabled the production of induced pluripotent stem cells (iPS) 

with variant mutations in NTCP in the genomic NTCP locus. 

Wild-type and variant NTCP iPS were differentiated into iHeps 

and then inoculated with HBV containing serum. NTCP variants 

had variable permissiveness for HBV infection. In particular the 

Ser267Phe mutation impacts NTCP expression and HBV permissiveness 

to HBV infection. These results exemplify the utility 

of a physiologically relevant infectious system for studying HBV 

interactions with relevant host cell genetics and physiology. 

Combining these systems with gene editing technologies will 

enable the dissection of clinically relevant mutations in physiologically 

relevant human infectious systems. 

Disclosures: 

The following authors have nothing to disclose: Angela Frankel, Robert E. 

Schwartz 

1681 

Genetic association study failed to confirm an association 

between the NTCP S267F mutation and persistence 

of hepatitis B virus or development of hepatocellular 

carcinoma in the Japanese population 










Background & Aims: Sodium taurocholate cotransporting 

polypeptide (NTCP) has long been known as a liver bile acid 

transporter but was also recently identified as a hepatocyte 

receptor for hepatitis B virus (HBV). The NTCP S267F mutation 

has been reported to reduce taurocholate transporting activity 

and result in defective HBV receptor function. This amino acid 

substitution corresponds to a single nucleotide polymorphism 

(SNP), rs2296651 is found in the Asian population but not in 

European, African and American populations, according to the 

1000 Genomes database. Moreover, this SNP frequency varies 

widely even among Asian populations: 12% (Chinese Dai), 

11% (Kinh (Vietnam)), 8% (Southern Han Chinese), 3% (Han 

Chinese), and 2% (Japanese). In this study, we investigated 

the importance of NTCP in HBV persistence and the development 

of HBV-induced hepatocellular carcinoma (HCC) by 

focusing on the S267F mutation using a large scale case-control 

association analysis. Methods: We genotyped SNPs in 

2,687 Japanese patients with chronic HBV infection (631 with 

HCC and 2,056 without HCC) and 5,489 control individuals 

without liver disease or cancer. Results: We found that minor 

allele (T) frequencies of rs2296651 were 0.013 and 0.010 in 

controls and chronic HBV patients, respectively. We observed 

3 minor allele homozygotes (TT genotype) in controls. In contrast, 

we did not observe any HBV patients with TT genotype. 

However, we found no significant differences in rs2296651 

frequencies between chronic HBV patients and controls in any 

genetic models. We also compared chronic HBV patients with 

and without HCC and found that their T allele frequencies were 

0.007 and 0.011, respectively. Finally, we found no significant 

association between the SNP genotype and development 

of HCC with or without adjustment for age and gender. These 

results might be partially affected by a low allele frequency 

of rs2296651. Therefore, we examined another NTCP SNP, 

rs4646287, which was reported as an intronic SNP associated 

with HBV-induced HCC susceptibility in the Han Chinese 

population and is more frequent in Japanese population than 

rs2296651. Previously reported risk allele (A) frequencies of 

rs4646287 were 0.144 and 0.142 in Japanese HBV patients 

with and without HCC, respectively, and we again found no 

significant association between SNP genotype and development 

of HCC. Conclusions: We found no genetic association 

between the S267F mutation of NTCP and HBV persistence or 

the development of HBV-induced HCC in the Japanese population. 

Further genetic analysis in populations with a higher 

frequency of rs2296651 would improve understanding about 

the role of NTCP in chronic HBV infection.


Disclosures: 













1682 

Serial Changes of Th1 and Th17 Cytokines and a SNP 

in HLA class II DPB1 Gene Associated with Hepatitis B 

Virus Reactivation in Patients Treated with Immunomodulatory 

Agents 

Hidetaka Matsuda, Tomoyuki Nemoto, Yoshihiko Ozaki, Tatsushi 

Naito, Masahiro Ohtani, Katsushi Hiramatsu, Hiroyuki Suto, 

Yasunari Nakamoto; Second Department of Internal Medicine, 

University of Fukui, Fukui, Japan 

BACKGROUND: Hepatitis B virus (HBV) reactivation can be 

triggered by various chemotherapies, in which the patients’ 

immunological status may be suppressive. As the patients’ 

immunity are influenced by human leukocyte antigen (HLA) 

class II mediated interactions between CD4 + helper T cells and 

antigen presenting cells, polymorphisms of HLA class II genes 

might affect the reactivation of HBV. In this study, we aimed to 

assess the cytokine levels reflecting immunosuppressive status 

and the candidate single nucleotide polymorphisms (SNPs) of 

HLA class II genes associated with the risks of HBV reactivation 

in patients treated with immunomodulatory therapies. METH- 

ODS: STUDY 1: Five patients with malignant lymphoma (ML) 

who undergone R-CHOP or CHOP and achieved complete 

remission (CR) in University of Fukui Hospital between 2013 

and 2014 were enrolled. The sera collected consecutively from 

the subjects were measured for a total of 27 cytokines, chemokines 

and growth factors by multiplex cytokine analysis. Immune 

profiles after the initiation of chemotherapies were investigated, 

and levels of the 27 factors were compared with those 

of 20 healthy controls. STUDY 2: A total of 43 patients with 

resolved HBV infection treated with immunosuppressive chemotherapies 

(e.g. R-CHOP, CHOP, methotrexate, infliximab, and 

cyclosporin) during 1999 to 2014 were investigated. All were 

followed up for more than 2 years; HBV had reactivated in 10 

of the 43 subjects, but not in the other 33. Genotyping of 22 

SNPs located within HLA class II DPA1, DPB1, DQA1, DQB1, 

and DRB1 genes was conducted by quenching probe PCR 

(geneCube). Genotype frequencies of the tested SNPs were 

compared between the two groups, whose latently infected 

HBV had reactivated and not. RESULTS: STUDY 1: In the tested 

patients with ML, serial levels of IL-17, IL-1β, IFN-γ, and G-CSF 

at the timing of CR with R-CHOP and CHOP were significantly 

decreased in contrast to those of the controls (123.1, 3.7, 

96.0, and 194.1 pg/ml vs. 158.2, 6.8, 146.2, and 306.5 

pg/ml, respectively; p 22.5% was 

significant higher than those with PD-1 expression increasing 

≤ 22.5% (46.15% vs 13.64%, P=0.033) from week 12 to 

week 24. Interestingly, the patients with both TLR2 expression 

< 18% at week 12 and PD-1 expression increasing > 22.5% 

from week 12 to week 24 had a significantly higher rate of 

virological breakthrough than the patients who met either of 

conditions mentioned above (83.33% vs. 9.09%, P=0.002), 

and the patients who failed to meet any condition (83.33% 

vs. 11.11%, P=0.005). Conclusions Downregulation of TLR2 

on CD14 + monocytes and upregulation of PD-1 on CD8 + T cells 

correlate with virological breakthrough during the treatment 

with Peg IFN alfa-2a after switching them from ETV therapy. 

Combination of TLR2 expression < 18% at week 12 and PD-1 

expression increasing > 22.5% from week 12 to week 24 has 

a high positive predictive value (PPV) for virological breakthrough 

in entecavir suppressed CHB patients with sequential 

Peg IFN alfa treatment.


Disclosures: 





The following authors have nothing to disclose: Di Wu, Weiming Yan, Guang 

Chen, Meifang Han 

1684 

IFN-α-mediated Base Excision Repair Pathway Correlates 

with Antiviral Response Against Hepatitis B Virus 

Infection 

Yong Li 1 , Yuchen Xia 3 , Meifang Han 2 , Guang Chen 2 , Wolfgang 

E. Thasler 4 , Xiaoping Luo 1 , Ulrike Protzer 3 , Qin Ning 2 ; 1 Tongji 

Hospital, Tongji Medical College, Huazhong University of Science 

and Technology, Wuhan, China; 2 Department and Institute 

of Infectious Diseases, Tongji Hospital, Tongji Medical College, 

Huazhong University of Science and Technology, Wuhan, China; 

3 Institute of Virology, TechnischeUniversität München–Helmholtz 

Zentrum München, Munich, Germany; 4 Department of General, 

Visceral, Ludwig Maximilians University, Munich, Germany 

Previous studies identified APOBEC family as base excision 

repair (BER) proteins with enzymatic activity through deamination 

of a cytidine base in DNA and/or RNA. As an 

enzyme, APOBEC is therefore part of an antiviral effector repertoire 

against hepatitis B virus (HBV) infection by interferon 

(IFN). Additionally, other BERs, namely NEIL3 and TDG, are 

expressed in liver and are regulated by IFN-α in hepatocytes. 

We thus hypothesized that the responses to IFN-α treatment of 

chronic hepatitis B (CHB) patients are relevant to IFN-induced 

deaminases and BER genes. Ten CHB patients treated with 

PEGylated IFN-α for 48 or 96 weeks, and six CHB-treatment 

naïve patients as controls were recruited. Response to IFN 

treatment was defined as HBV DNA 1 log10IU/ 

ml. Blood and liver samples were collected, and APOBEC3 

and other BER genes measured by real-time PCR. The correlations 

between BER gene expression levels and IFN treatment 

responses were studied in these patients as well as in primary 

human hepatocytes (PHH) and terminally differentiated HepRG 

cells in vitro. Compared to treatment-naive patients, APO- 

BEC3-A,-B, -C, -DE, and -G mRNA levels were up-regulated in 

IFN-treated subjects. APOBEC3-A was significantly increased 

in IFN-treated responders than non-responders. In contrast, 

other BER genes, NEIL3 and TDG, were down-regulated in both 

IFN-treated patients and in IFN-treated PHH and HepRG cells. 

APOBEC3 and BER gene expression at treatment endpoints 

partially correlated with the corresponding degree of HBsAg/ 

HBV DNA decline and DNA level. Our study suggests that the 

expression levels of editing enzymes APOBEC3-A, -C, -F, -G, 

and NEIL3 and TDG correlates with IFN treatment responses in 

CHB patients, which may serve as biomarkers for CHB disease 

management. 

Disclosures: 

Ulrike Protzer - Advisory Committees or Review Panels: GILEAD, Roche, MedImmune; 

Board Membership: University Hospital Cologne; Grant/Research Support: 

Janssen, Roche 





The following authors have nothing to disclose: Yong Li, Yuchen Xia, Meifang 

Han, Guang Chen, Wolfgang E. Thasler, Xiaoping Luo 

1685 

Hepatitis B virus down regulates the expression levels of 

Vitamin D receptor in hepatocellular carcinoma (HepG2) 

transfected cells thereby preventing the inhibition of 

viral transcription and production by Vitamin D 

Neta Gotlieb 1 , Irina Tachlytski 1 , Maya Sultan 1 , Michal Safran 1 , 

Ziv Ben Ari 1,2 ; 1 Liver Disease Center, Sheba Medical Center, Tel 

Hashomer, Israel; 2 The Sackler School of Medicine, Tel Aviv University, 

Tel-Aviv, Israel 

Vitamin D is an important immune modulator that plays an 

emerging role in both the innate and adaptive immune systems. 

Certain viruses as the HIV-1 are capable to impair the innate 

immune defense by down regulating the vitamin D receptor 

(VDR) pathway. Recently it has been demonstrated that low 

25(OH)D3 serum levels are associated with high levels of hepatitis 

B Virus (HBV) replication in patients with HBV infection. 

Our aim was to study in vitro the relationship between HBV 

transcription and production and Vitamin D signaling pathway 

and to explore the associated mechanism(s). Methods: To measure 

HBV transcription and production we used qRT-PCR on 

RNA purified from HBV-transfected hepatocellular carcinoma 

HepG2 cells (HepG2 215) using primers specific to RNA- 

HBV. In addition, the level of HBV replication was evaluated 

by determining the concentrations of HBsAg levels in the cell 

culture medium of HepG2 215 using specific enzyme-linked 

immunosorbent (Elisa) assay. cccDNA levels were quantified 

using qRT-PCR on DNA extracted from HepG2 215 cells. The 

level of HBV transcription and production was assessed with 

and without the administration of vitamin D (0-200mM) or its 

active metabolite calcitriol (0-100 nM) for 48 or 72 hours. VDR 

and IFNβ transcripts expression level were assessed by qRT- 

PCR. Levels were compared in RNAs purified from HepG2 cells 

(control) versus HepG2 215. Results: The expression level of 

VDR transcript in HepG2 215 cells was statistically much lower 

compared with non-transfected HepG2 cells. Furthermore, the 

administration of vitamin D or calcitriol did not suppress the 

secretion of HBsAg, the cccDNA expression and the HBVRNAs 

level in HepG2 215 compared to cells which were not treated 

with Vitamin D. Increasing the dose of both vitamin D and 

calcitriol did not affect these findings. In addition, the administration 

of Vitamin D or calcitriol induced interferon signaling 

pathway, resulting in up-regulation of IFNβ expression level in 

HepG2 cells, however, no such increase was observed in the 

HepG2 215 HBV transfected cells. Conclusions: Our results 

indicate for the first time that VDR expression level is down-regulated 

in HBV-transfected HepG2 cells thereby preventing vitamin 

D to affect transcription and production of the HBV in 

vitro. Furthermore, these findings suggest that HBV might use 

this mechanism to avoid the immunological defense system by 

affecting the interferon signaling pathway and the expression 

of the IFNβ. 

Disclosures: 

The following authors have nothing to disclose: Neta Gotlieb, Irina Tachlytski, 

Maya Sultan, Michal Safran, Ziv Ben Ari 

1686 

The role of HBV quasispecies evolution in HBeAg-negative 

reactivation 

Guan Huei Lee 1,2 , Hui Heng Chong 2 , Seng Gee Lim 2 ; 1 Medicine, 

National University Health System, Singapore, Singapore; 2 Medicine, 

Yong Loo Lin School of Medicine, Singapore, Singapore 

Background: HBeAg negative chronic hepatitis B reactivation is 

characterized by increased levels of HBV DNA accompanied 

by abnormal ALT, and occasionally severe hepatitis B flare.


We have shown previously that viral quasispecies evolution 

and diversity may play a pathogenic role in development of 

HBeAg seroconversion. However, the role of viral quasispecies 

in pathophysiology of reactivation in HBeAg-negative chronic 

hepatitis B is uncertain. Methods: Twenty-two HBeAg-negative 

chronic hepatitis B patients who had serial serum samples 

collected before and after HBV reactivation were analysed. 

Full length HBV genomes were amplified from serum extracted 

DNA and were cloned into pCR®2.1-TOPO vector. At least 

20 positive clones were randomly selected and sequenced 

using Big Dye Terminator. The samples were analyzed for point 

mutation in precore stop codon, core promoter- Enhancer II, S1 

promoter, S2 promoter and Enhancer I- X promoter regions. 

Ten inactive HBV carriers were also analysed as controls. Fulllength 

HBV genome collected before and after reactivation 

were transfected into Huh7 cells for functional analysis of their 

replication fitness. Results: In subjects with clear clinical reactivation 

profile, clonal selection was demonstrated (9 out of 

10 subjects) by DNA phylogenetic analysis (p=0.005). The 

genetic diversity and viral evolution rates were not statistically 

different from the inactive controls except except at the precore/core 

ORF (p=0.035, Mann-Whitney U test). 80% of reactivation 

patients had precore stop codon G to A substitution at 

nucleotide position 1896. In addition, other mutations in the 

promoter/enhancer regions were identified with more than fifty 

percent difference between time point 1 and time point 2 (and 

not found in control subjects). HBV clones isolated during HBV 

reactivation showed higher HBV DNA and HBsAg production 

rates than the clones isolated during inactive phase of the same 

patient in transfected Huh7 cell line. Conclusions: Significant 

clonal selection occurred during HBeAg-negative reactivation. 

Precore stop codon substitution G1896A occurs significantly 

in majority of the patients. The nucleotide variability in the precore 

promoter, and possibly other promoter/enhancer regions 

may play an important role in the progression of HBV disease, 

although more functional analyses of individual mutations are 

required. 

Disclosures: 






The following authors have nothing to disclose: Guan Huei Lee, Hui Heng Chong 

1687 

Hepatic macrophage IL-1 beta expression and bile and 

sterol transporter suppression precede cholestasis in a 

parenteral nutrition mouse model 

Karim C. El Kasmi, Aimee Anderson, Michael W. Devereaux, 

Ronald J. Sokol; Pediatrics, UC Denver, Aurora, CO 

Background: Parenteral Nutrition Associated Cholestasis 

(PNAC) is a serious complication of long term PN infusion in 

children and adults with intestinal failure. We have previously 

reported in a PNAC mouse model that activation of hepatic 

macrophages and up-regulation of pro-inflammatory cytokines 

(IL-1 beta) was associated with suppression of hepatic bile 

(Abcb11/BSEP, Abcc2/MRP2) and sterol (ABCG5/8/sterolin) 

transporters. IL-1R-/- mice were protected from PNAC, with 

normalization of hepatic transporter expression, supporting 

a causal role for IL-1 beta. The objective of this study was 

to determine the role of IL-1 beta at early stages of PNAC 

by examining the temporal relationship at early time points 

between IL-1 beta, transporter expression, and cholestasis. 

Methods and Results: PNAC model: wild type C57/B6 mice 

were exposed to dextran sulfate sodium (DSS) in drinking 

water (to induce intestinal injury) for 4 days followed by infusion 

of soy lipid emulsion-based PN through a central venous 

catheter for 3 or 7 days (DSS-PN mice); appropriate controls 

were conducted. Cholestasis (increased serum bile acids and 

bilirubin) and hepatocyte injury (increased AST and ALT) 

were absent in DSS/PN3d mice but developed in all DSS/ 

PN7d mice. IL-1 beta transcription was significantly induced 

in liver homogenate and in isolated hepatic mononuclear cells 

(hMNCs) at 3 days in DSS/PN mice and persisted at 7 days. 

Concomitantly, transcription of Abcb11, Abcc2, and Abcg5/8 

was suppressed at both of these time points in DSS/PN mice, 

indicating that macrophage activation and transporter suppression 

preceded the onset of biochemical cholestasis. To further 

determine if IL-1 beta was responsible for the transcriptional 

suppression, wild type mice were injected i.p. with recombinant 

IL-1 beta (200ng/mouse) and sampled after 4 hrs, and 

HuH7 and HepG2 cells (human hepatocyte cell lines) were 

incubated with IL-1b (10-50ng/ml) for 4 hrs, and gene expression 

measured. IL-1 beta treatment significantly suppressed 

expression of Abcb11, Abcc2, and Abcg5/8 in the mice and 

in both hepatocyte cell lines. Incubation of these cell lines with 

LPS (100-1000ng/ml) had no effect on these transporters. Conclusions: 

Activation of hepatic macrophages and generation of 

IL-1 beta, as well as suppression of bile and sterol transporters, 

occurred after only 3 days of PN infusion and preceded onset 

of cholestasis. Furthermore, IL-1 beta was sufficient to suppress 

these transporters in cultured hepatocytes and in vivo. These 

data support IL-1 beta as a critical early mediator in the pathogenesis 

of PNAC and suggest IL1-beta signaling as a potential 

therapeutic target in this disorder. 

Disclosures: 

Ronald J. Sokol - Advisory Committees or Review Panels: Yasoo Health, Inc.; Consulting: 

Roche, Ikaria, Otsuka American Pharmaceuticals, Alnylam, Retrophin; 

Grant/Research Support: Mead Johnson Nutritionals, Lumena, FFF Enterprises 

The following authors have nothing to disclose: Karim C. El Kasmi, Aimee Anderson, 

Michael W. Devereaux 

1688 

Molecular mechanism of IL-1 beta mediated transcriptional 

suppression of the sterol transporter Abcg5/8 in 

a parenteral nutrition mouse model 

Karim C. El Kasmi, Aimee Anderson, Padade M. Vue, Michael 

W. Devereaux, Natarajan Balasubramaniyan, Frederick J. Suchy, 


Background: Parenteral nutrition associated cholestasis (PNAC) 

is the leading indication for combined intestine/liver transplant. 

A possible role of intravenous soy lipid emulsions in PNAC 

pathogenesis has led to concern about the hepatic toxicity of 

phytosterols contained in soy lipid emulsions. We have demonstrated 

in a novel PNAC mouse model that soy lipid emulsions 

in PN are associated with induction of hepatic macrophage 

IL1b expression concomitant with accumulation of hepatic phytosterols 

and transcriptional suppression of hepatocyte Abcg5/ 

g8, coding for the canalicular transporter of sterols, as well 

as BSEP and MRP2. The aim of this study was to elucidate the 

role of IL1b signaling in suppression of ABCG5/8 transcription 

during PNAC. Methods and Results: Wild type C57/B6 mice 

were exposed to dextran sulfate sodium (DSS) (to induce intestinal 

injury) for 4d followed by infusion of phytosterol-containing 

(soy lipid) PN solution through a central venous catheter for 14 

d (DSS-PN mice) and developed cholestasis (increased serum 

bile acids and bilirubin) and hepatocyte injury (increased AST 

and ALT). DSS-PN mice had hepatic IL1b induction and markedly 

reduced hepatic mRNA for Abcb11, Abcc2, Abcg5/8 

compared to control mice. To further elucidate the role of IL1b 

in this transcriptional suppression, mice with genetic deletion


of the receptor for IL-1 (IL1R-/-) were exposed to DSS-PN for 

14d. DSS-PN/IL1R-/- mice had significantly reduced serum 

AST, ALT, bile acids, and bilirubin compared to DSS-PN mice. 

Hepatic gene mRNA for Abcb11, Abcc2, and Abcg5/8 was 

not reduced in DSS-PN IL1R-/- mice. Chromatin-immuno-precipitation 

assays (ChIP) on liver demonstrated that in WT DSS-PN 

mice binding of NFkB (downstream of IL1 signaling) to the 

Abcg5/8 promoter was increased concomitant with reduced 

binding of LXR (the transcription factor that promotes Abcg5/8 

expression). In contrast, in DSS-PN IL1R-/- mice, binding of 

NFkB was reduced concomitant with increased binding of LXR 

to the Abcg5/8 promoter. Supporting these in vivo studies, 

incubation of HepG2 and HuH7 cells with IL1b significantly 

suppressed Abcg5/8 mRNA while markedly increasing NFkB 

reporter activity. Finally, incubation of HepG2 cells with IL1b 

suppressed the ability of an LXR agonist to induce activity 

ABCG5/8 promoter linked luciferase. Conclusion: Hepatic IL1b 

is a critical mediator in the pathogenesis of PNAC by promoting 

suppression of the phytosterol transporter Abcg5/8 through 

increased binding of NFkB and decreased binding of LXR to 

the Abcg5/8 promotor. Reduced Abcg5/8 causes accumulation 

of phytosterols which then interfere with hepatocyte FXR 

signaling, thus promoting cholestasis. 

Disclosures: 





Padade M. Vue, Michael W. Devereaux, Natarajan Balasubramaniyan, 

Frederick J. Suchy 

1689 

Enteral Obeticholic Acid Promotes Intestinal Growth in 

Total Parenteral Nutrition Fed Neonatal Pigs 

Yanjun Jiang 1 , Zhengfeng Fang 2 , Barbara Stoll 1 , Gregory J. Guthrie 

1 , Jens Holst 3 , Bolette Hartmann 3 , Douglas Burrin 1,4 ; 1 USDA 

Children’s Nutrition Research Center, Department Pediatrics, Baylor 

College of Medicine, Houston, TX; 2 Animal Nutrition Institute, 

Sichuan Agricultural University, Chengdu, China; 3 The NNF Center 

for Basic Metabolic Research and the Department of Biomedical 

Sciences, University of Copenhagen, Copenhagen, Denmark; 

4 Pediatric Gastroenterology, Hepatology and Nutrition, Department 

Pediatrics, Baylor College of Medicine, Houston, TX 

Intestinal atrophy is an adverse outcome associated with prolonged 

total parenteral nutrition (PN) partly due to disruption 

of normal enterohepatic circulation of bile acids. Previously 

we showed that enteral treatment with chenodeoxycholic acid 

(CDCA), a dual agonist for the nuclear receptor, farnesoid X 

receptor (FXR) and G protein-coupled receptor TGR-5, induced 

intestinal mucosal growth in a parenteral nutrition associated 

liver disease (PNALD) piglet model. We hypothesized that the 

intestinal trophic CDCA effects were mainly mediated by TGR5 

receptor-mediated glucagon-like peptide 2 (GLP-2) released 

from enteroendocrine cells. However, CDCA may also exert 

trophic effects via FXR signaling in intestine. The aim of the current 

study was to compare the physiological effects of a selective 

and potent FXR agonist, obeticholic acid (OCA) vs CDCA 

on intestinal growth in TPN-fed pigs. Term, newborn pigs were 

assigned to receive complete TPN (PN), PN + enteral CDCA 

(30 mg/kg), or PN +enteral OCA (0.5, 5, 15 mg/kg) daily for 

19 d. The daily parenteral lipid was Intralipid given at 10 g/ 

kg. Intestinal growth and crypt cell proliferation (in vivo BrdU 

labeling) were measured. We found that both CDCA and OCA 

treatments significantly increased small intestinal weight, compared 

to PN pigs, but OCA15 was higher than CDCA (146% 

vs. 118%). The OCA-induced increase in jejunal and Ileal 

weight was dose-dependent, yet the trophic effects of OCA and 

CDCA were greater in the ileum than jejunum. Ileal villus height 

and crypt depth were increased by OCA and CDCA. The percentage 

of BrdU positive crypt cells in PN, CDCA, OCA0.5, 

OCA5 and OCA15 groups were 31%, 40%, 40%, 46% and 

46% respectively, suggesting OCA and CDCA increased crypt 

cell proliferation. Portal plasma GLP-1 and -2 concentrations 

were significantly increased in pigs treated with CDCA, but not 

OCA, compared to PN pigs suggesting differential intestinal 

activation of TGR5 signaling. OCA, but not CDCA, treatment 

dose-dependently increased ileal transcriptional expression of 

FXR target genes, small heterodimer partner (SHP), ileal lipid 

binding protein (ILBP), and fibroblast growth factor 19 (FGF19). 

The expression of fibroblast growth factor receptor 4 (FGFR4) 

and β-Klotho mRNA were relative abundant in ileal tissue in all 

groups. We conclude that the intestinal trophic effects of OCA 

are greater than CDCA in TPN-fed pigs. The trophic effects of 

CDCA appear to occur via TGR5-mediated GLP-2 secretion 

rather than FXR signaling. We show novel evidence that OCA 

exerts trophic effects in the neonatal intestine and this appears 

to act mainly via FXR-dependent mechanisms. 

Disclosures: 

The following authors have nothing to disclose: Yanjun Jiang, Zhengfeng Fang, 

Barbara Stoll, Gregory J. Guthrie, Jens Holst, Bolette Hartmann, Douglas Burrin 

1690 

Human iPSC-derived hepatocyte-like cells generated 

from patients with bile salt export pump deficiency 

recapitulate the phenotype of progressive familial intrahepatic 

cholestasis type 2 

Kazuo Imagawa 1,2 , Kazuo Takayama 2,3 , Shigemi Isoyama 1 , Ken 

Tanikawa 4 , Masato Shinkai 5 , Masayoshi Kage 4 , Kenji Kawabata 

6,7 , Ryo Sumazaki 1 , Hiroyuki Mizuguchi 2,3 ; 1 Department of 

Child Health, Faculty of Medicine, University of Tsukuba, Tsukuba, 

Japan; 2 Laboratory of Hepatocyte Regulation, National Institute of 

Biomedical Innovation, Osaka, Japan; 3 Laboratory of Biochemistry 

and Molecular Biology, Graduate School of Pharmaceutical 

Sciences, Osaka University, Osaka, Japan; 4 Department of 

Diagnostic Pathology, Kurume University Hospital, Kurume, Japan; 

5 Department of Surgery, Kanagawa Children’s Medical Center, 

Yokohama, Japan; 6 Laboratory of Stem Cell Regulation, National 

Institute of Biomedical Innovation, Osaka, Japan; 7 Laboratory of 

Biomedical Innovation, Graduate School of Pharmaceutical Sciences, 

Osaka, Japan 

Background: The bile salt export pump (BSEP) is a key molecule 

that transports bile acid into the biliary canaliculi in 

humans. Progressive familial intrahepatic cholestasis type 2 

(PFIC2) is mainly characterized by BSEP deficiency, which is 

followed by intrahepatic cholestasis. In many cases, liver transplantation 

is the most effective therapy available. Hence, it is 

necessary to identify novel therapeutic alternatives. To clarify 

the underlying disease mechanisms and discover the novel 

therapeutic options, we generated iPSCs from BSEP deficient 

(BD) patients and analyzed the differentiated hepatocyte-like 

cells (HLCs). Methods: One healthy donor and two BD patients 

participated in present study. Patient 1 was diagnosed with 

normal-gamma glutamyl transferase PFIC and presented with 

the PFIC2 phenotype in infancy. Although her clinical features, 

liver histology results, and BSEP deficiency were indicative 

of the PFIC2 phenotype, exonic mutations were not found in 

the BSEP gene. Patient 2 was diagnosed with PFIC2 based 

on the presence of compound heterozygous mutations in the 

BSEP gene (c.-24C>A and c.2416G>A) and liver histology 

results. The iPSCs were generated from blood cells obtained 

from all three participants (Control-iPSC and BD-iPSC) by using


a Sendai virus vector expressing Yamanaka factors. To examine 

whether these iPSCs expressed pluripotent markers, realtime 

RT-PCR and immunostaining analyses were performed. 

The iPSCs were then differentiated into hepatocyte-like cells. 

To examine the BSEP expression levels in HLCs derived from 

BD-iPSC (BD-iPS-HLCs) and their biliary excretion capacity, we 

performed real-time RT-PCR, immunostaining analyses, and 

cholyl-lysyl-fluorescein (CLF) excretion experiments. Results: We 

successfully obtained iPSCs from blood cells. These cells were 

positive for OCT4 and expressed markers specific to human 

embryonal stem cells. The HLCs were obtained on day 25 after 

hepatocyte differentiation. Bile canaliculi between the cells 

were observed by electron microscopy. BSEP was expressed 

in the membranes of Control-iPSC derived HLCs (Control-iPS- 

HLCs), while BD-iPS-HLCs did not show localized expression of 

BSEP in its membranes. The accumulation of CLF in bile canaliculi 

was observed in Control-iPS-HLCs, but not in BD-iPS-HLCs, 

suggesting that the biliary excretion capacity was impaired 

in BD-iPS-HLCs. Thus, the data implied that BD-iPS-HLCs could 

reproduce the pathophysiology of PFIC2. Conclusions: BD-iPS- 

HLCs recapitulated the PFIC2 phenotype, mislocalization of 

BSEP, and impairment of biliary excretion. Thus, this PFIC2 

model can be applied to elucidate disease mechanisms and to 

determine novel therapeutic options. 

Disclosures: 

The following authors have nothing to disclose: Kazuo Imagawa, Kazuo 

Takayama, Shigemi Isoyama, Ken Tanikawa, Masato Shinkai, Masayoshi Kage, 

Kenji Kawabata, Ryo Sumazaki, Hiroyuki Mizuguchi 

1691 

King’s-Variceal Prediction Score (K-VaPS); a non invasive 

assessment tool of severe portal hypertension in 

cirrhotic children 

Peter Witters, Dominic A. Hughes, Palaniswamy Karthikeyan, 

Alastair J. Baker, Mark Davenport, Anil Dhawan, Tassos Grammatikopoulos; 

Paediatric Liver GI & Nutrition Centre, King’s College 

Hospital NHS Foundation Trust, London, United Kingdom 

Background & aim: Variceal hemorrhage is a serious complication 

of chronic liver disease(CLD) in children. There is limited 

knowledge around the best prophylactic management and 

selection criteria of children undergoing surveillance endoscopy 

(OGD). We derive a new prediction model of clinically 

significant varices (CSV) and validate it in a separate cohort. 

Methods: All patients presenting in our centre with suspected 

PHT or gastrointestinal (GI) bleeding due to CLD and undergoing 

a first OGD between January 2005- December 2012 were 

included. A validation cohort from 2013-2015 was obtained. 

Clinical, biochemical and radiological data were collected. 

Results: Data on 124(67M) treatment-naïve patients were collected. 

Mean age was 8.81 ± 5.60 years. 50% had biliary 

atresia and the remainder various other CLD types. 35(28%) 

presented with GI bleeding and overall 79(64%) were in 

CSV+ve. Mean values in CSV+ve vs. CSV-ve group were for 

haemoglobin(Hb)(mg/dL) [10,58 vs. 11,62 p=0.008], platelet 

count(10 9 /ml) [111 vs. 167, p=0.001], white cell count(10/ 

ml) [5.3 vs. 6.9, p=0.019], INR[1.22 vs. 1.13, p=0.004], bilirubin(mg/dL) 

[53.33 vs. 28.22, p=0.014], albumin(g/L) [37 

vs. 41, p=0.0001], AST(IU/L) [121 vs. 104, p=0.815], spleen 

size(cm) [16.41 vs. 14.49, p=0.073], spleen size(z-score) 

[9.1 vs. 6.35, p=0.009] and equivalent adult spleen size(E- 

ASS) (cm) [23.21 vs. 19.3, p=0.003], respectively. Previously 

published predictions rules had, at optimal cut-off, a sensitivity 

and specificity of 76% and 59%(CPR), 60% and 55%(APRI), 

80% and 59%(VPR), respectively. To construct a new prediction 

score Pearson’s bivariate correlation was performed. 

Variables significantly correlating with CSV were Hb(R=-.250, 

p= 0.005), platelet count(R=-.276, p=0.002), INR(R=0.223, 

p=0.010), albumin(R=-.334, p


Th1 cytokines (TNF-α and IFN-γ) and innate cytokines (IL-1, IL-6) 

in WT-RRV mice but not BSS controls or Ig-α -/- RRV groups. Conclusions: 

The RRV-induced mouse model of BA is associated 

with an abnormal decrease in the expression of the majority 

of transporters involved in bile excretion. The high levels of 

retained bile acids may contribute to liver injury. The lack of 

inflammation within the Ig-α -/- RRV mice is associated with normal 

expression of the majority of bile acid transporter proteins, 

suggesting that the inflammation present in RRV-induced BA 

mice inappropriately inhibits bile acid transporter expression. 

Disclosures: 

The following authors have nothing to disclose: Asokan Rengasamy, Karim C. 

El Kasmi, Brianna Traxinger, Jonathan Roach, Natarajan Balasubramaniyan, 

Cara L. Mack 

1693 

Exome Sequencing and de novo Mutation Analysis 

Reveal a Highly Connected Gene Network in Isolated 

Biliary Atresia 

Kathleen M. Loomes 5,1 , Ramakrishnan Rajagopalan 2 , Christopher 

Grochowski 2 , Ying Chen 3,4 , Melissa Gilbert 2 , Henry C. Lin 5,1 , Marcella 

Devoto 4,1 , Nancy B. Spinner 2,6 ; 1 Department of Pediatrics, 

University of Pennsylvania Perelman School of Medicine, Philadelphia, 

PA; 2 Department of Pathology and Laboratory Medicine, The 

Children’s Hospital of Philadelphia, Philadelphia, PA; 3 Genomics 

and Computational Biology Graduate Group, University of Pennsylvania 

Perelman School of Medicine, Philadelphia, PA; 4 Division 

of Human Genetics, The Children’s Hospital of Philadelphia, 

Philadelphia, PA; 5 Division of Gastroenterology, Hepatology and 

Nutrition, The Children’s Hospital of Philadelphia, Philadelphia, 

PA; 6 Department of Pathology and Laboratory Medicine, University 

of Pennsylvania Perelman School of Medicine, Philadelphia, PA 

Biliary atresia (BA) is a severe pediatric liver disease resulting 

in necroinflammatory obliteration of the extrahepatic biliary 

tree. The majority of BA patients develop biliary cirrhosis and 

50% will require liver transplantation in the first two years of 

life. Incidence of BA is reported to be between 1 in 18,000 

and 1 in 8,000 depending on ethnicity. The etiology of BA 

is unknown, with evidence for infectious, environmental, and 

genetic risk factors described. The purpose of this study was to 

investigate possible genetic etiologies of BA by whole exome 

sequencing of non-syndromic BA probands and their parents 

(trios). DNA samples were obtained from 10 well-characterized 

non-syndromic BA probands and their parents from the 

NIDDK-funded Childhood Liver Disease Research Network 

(ChiLDReN). We performed whole-exome sequencing in these 

10 trios and looked for de novo mutations under the assumption 

that mutations only seen in probands, but not the parents, 

are more likely to be involved in the etiology of the disease. 

We used a custom in-house computational analysis pipeline to 

call variants and identify de novo mutations. We found a total 

of 33 exonic de novo variants in 33 different genes. These 

were 2 loss-of-function mutations, 3 in-frame insertions/deletions, 

2 splice-site mutations and 26 missense mutations. The 

number of de novo changes per proband ranged from 1 to 9 

with a mean of 3.3. Seven of the 10 probands carried more 

than 1 de novo mutation. However, none of the mutations was 

seen in more than one proband. We used protein-interaction 

network analysis tools such as GeneMANIA, DAPPLE and Ingenuity 

Pathway Analysis (IPA) to assess 1) whether the genes 

with de novo mutations are connected with each other directly 

or through a single common partner, 2) whether the genes 

with de novo mutations form a network with genes previously 

implicated in BA. Gene network analysis with GeneMANIA 

suggested a highly interconnected network of proteins that 

are co-expressed. Protein interaction analysis with DAPPLE 

suggested that 17 out of 33 genes (51%) interact with each 

other directly or through a single common partner to form a 

tight network. We used IPA to explore the paths between the 

genes with de novo mutations and genes previously implicated 

in BA. Network connectivity analyses with IPA suggest that 

29 out of 33 genes (88%) are at most 2 nodes away from a 

group of 19 selected BA candidate genes. Results from network 

analysis suggest a shared etiology to seemingly sporadic de 

novo mutations observed in probands with isolated BA. To our 

knowledge, this is the first report of trio analysis for isolated BA 

using next-generation sequencing technology. 

Disclosures: 

The following authors have nothing to disclose: Kathleen M. Loomes, Ramakrishnan 

Rajagopalan, Christopher Grochowski, Ying Chen, Melissa Gilbert, Henry 

C. Lin, Marcella Devoto, Nancy B. Spinner 

1694 

DXA Bone Density in Alagille Syndrome Correlates with 

Fracture History and Degree of Cholestasis 

Kathleen M. Loomes 2 , Cathie Spino 14 , Nathan P. Goodrich 1 , 

Thomas N. Hangartner 15 , Amanda E. Marker 15 , James E. Heubi 16 , 

Binita M. Kamath 13 , Benjamin Shneider 17 , Philip Rosenthal 3 , 

Paula M. Hertel 17 , Saul J. Karpen 4 , Nanda Kerkar 5,19 , Jean P. 

Molleston 6 , Karen F. Murray 7 , Kathleen B. Schwarz 8 , Jeffrey Teckman 

18 , Yumirle P. Turmelle 9 , Peter F. Whitington 20 , Averell H. 

Sherker 10 , John C. Magee 11 , Ronald J. Sokol 12 ; 1 Arbor Research 

Collaborative for Health, Ann Arbor, MI; 2 Pediatric Gastroenterology, 

Hepatology and Nutrition, Children’s Hospital of Philadelphia, 

Philadelphia, PA; 3 Division of GI, Hepatology and Nutrition, 

UCSF-University of California, San Francisco, San Francisco, CA; 

4 Pediatric Gastroenterology, Hepatology and Nutrition, Emory 

University School of Medicine/Children’s Healthcare of Atlanta, 

Atlanta, GA; 5 Children’s Hospital of Los Angeles, Los Angeles, CA; 

6 Pediatric Gastroenterology, Hepatology and Nutrition, Indiana 

University School of Medicine/Riley Hospital for Children, Indianapolis, 

IN; 7 Division of Gastroenterology and Hepatology, University 

of Washington Medical Center, Seattle Children’s, Seattle, 

WA; 8 Johns Hopkins School of Medicine, Baltimore, MD; 9 Washington 

University School of Medicine, Saint Louis, MO; 10 Liver Disease 

Research Branch, National Institute of Diabetes and Digestive 

and Kidney Diseases, National Institutes of Health, Bethesda, MD; 

11 University of Michigan Medical School, Ann Arbor, MI; 12 Section 

of Pediatric Gastroenterology, Hepatology and Nutrition, 

Department of Pediatrics, University of Colorado, Aurora, CO; 

13 Division of Gastroenterology, Hepatology and Nutrition, Hospital 

for Sick Children and University of Toronto, Toronto, ON, Canada; 

14 University of Michigan, Ann Arbor, MI; 15 Department of 

Biomedical, Industrial, & Human Factors Engineering, Wright State 

University, Dayton, OH; 16 Division of Pediatric Gastroenterology, 

Hepatology and Nutrition, Cincinnati Children’s Hospital Medical 

Center, Cincinnati, OH; 17 Pediatric Gastroenterology, Hepatology 

and Nutrition, Baylor College of Medicine, Houston, TX; 18 Saint 

Louis University, Cardinal Glennon Children’s Medical Center, 

St. Louis, MO; 19 Mount Sinai, New York, NY; 20 Department of 

Pediatrics, Ann & Robert H. Lurie Children’s Hospital of Chicago, 

Northwestern University Feinberg Medical School, Chicago, IL 

Osteopenia and bone fractures (fx) are significant causes of 

morbidity in children with cholestatic liver disease. We performed 

dual energy X-ray absorptiometry (DXA) analysis in 

a cohort of children with inherited cholestatic diseases and 

explored associations with anthropometrics, laboratory measurements 

and fx history. Methods: Subjects were enrolled 

in the Longitudinal Study of Genetic Causes of Intrahepatic 

Cholestasis (LOGIC) in the NIDDK-funded Childhood Liver


Disease Research Network (ChiLDReN). DXA was performed 

on subjects age > 5 years (with native liver), diagnosed with 

Bile Acid Synthetic Disorder (BAD; n=14), Alpha-1 antitrypsin 

deficiency (A1AT; n=44) and Alagille syndrome (ALGS; 

n=49). Pearson correlation coefficients examined associations 

between subject reference (adjusted for age, gender and race) 

and anthropometrically-adjusted DXA Z-scores and laboratory 

values (total bilirubin (TB), serum bile acids (SBA) and 25-OH 

vitamin D (vitD)) and fx history. Results: Weight, height and 

BMI Z-scores were all lower, and TB and SBA higher, in the 

ALGS group (p1 fx; 21% and 7% in BAD; 

20% and 2% in A1AT). Bone mineral density (BMD) reference 

measures did not differ among disease groups except for 

total hip BMD and femoral neck BMD, which were lower in 

ALGS (p=0.007 and p=0.0002). Total body and total body 

minus head bone mineral content (BMC) reference measures 

were lower in the ALGS subjects (p


1696 

Diagnostic determination system for high-risk screening 

for inborn errors of bile acid metabolism: Results during 

20 years in East Asia 

Nakayuki Naritaka 1 , Mitsuyoshi Suzuki 1 , Hajime Takei 2 , Akihiko 

Kimura 3 , Takao Kurosawa 4 , Takashi Iida 5 , Hiroshi Nittono 

2 , Toshiaki Shimizu 1 ; 1 Pediatrics, Juntendo University Faculty 

of Medicine, Tokyo, Japan; 2 Junshin Clinic bile acid institute, 

Tokyo, Japan; 3 Pediatrics, Kurume University School of Medicine, 

Fukuoka, Japan; 4 Faculty of Pharmaceutical Sciencies, Health Services 

University of Hokkaido, Hokkaido, Japan; 5 Nihon University 

College of Humanities and Sciences, Tokyo, Japan 

BACKGROUND: Some patients with cholestasis of unknown 

cause may have inborn errors of bile acid metabolism (IEBAM), 

leading to abnormalities of bile acid biosynthesis. Although 

many types of bile acid synthesis defects have been reported 

for this disorder, no detailed information on its incidence and 

other aspects in East Asia has been available. To elucidate the 

current status of IEBAM, in July 1996, a diagnostic determination 

system was established for high-risk screening for IEBAM. 

METHODS: The target patients included children with hepatic 

disorders for which the cause could not be identified on conventional 

liver function testing or hepatitis virus testing, patients 

with liver cirrhosis of unknown etiology, sibling cases, and 

patients with cholestasis who exhibited serum levels of direct 

bilirubin (D-Bil) was ^2.0mg/dL,besides total bile acid and 

γ-glutamyltranspeptidase (GGT) were within the normal range. 

Urine samples were sent to the Bile Acid Institute which located 

in Tokyo, via refrigerated delivery service or impregnated filter 

paper with sufficient urine volume. The samples were analyzed 

during the 20 years between July 1996 and May 2015.Urinary 

bile acids were analyzed via gas chromatography–mass 

spectrometry (GC/MS) or liquid chromatography-tandem mass 

spectrometry (LC-MS/MS). RESULTS: In 960 cases analyzed 

over a 20-year period, 12 samples were differentially diagnosed 

with IEBAM, including: 3β-hydroxy-Δ 5 -C 27 

-steroid dehydrogenase/isomerase 

deficiency in 5 (2 in Japan, 2 in China, 

1 in Thailand), 3-oxo-Δ 4 -steroid 5β-reductase deficiency in 3 

(all in Japan), oxysterol 7α-hydroxylase deficiency in 3 (1 in 

Japan, 2 in Taiwan), bile acid-CoA amino acid N-acyltransferase 

deficiency in 1 (speculated in Thailand). Especially in 

Japanese cases, 2 cases each of 3β-hydroxy-Δ 5 -C 27 

-steroid 

dehydrogenase/isomerase deficiency and 3-oxo-Δ 4 -steroid 

5β-reductase deficiency were effectively treated with oral bile 

acid therapy. The third case of 3-oxo-Δ 4 -steroid 5β-reductase 

deficiency had spontaneous remission without oral therapy. The 

Japanese case of oxysterol 7α-hydroxylase deficiency underwent 

successful heterozygous living donor liver transplantation. 

CONCLUSIONS: Twelve patients were identified with cholestasis 

of unknown cause in East Asia during 20 years. IEBAM is 

a rare hereditary disease; therefore, much cost and labor is 

spent to make a definitive diagnosis via genetic analysis. The 

diagnostic determination system with urinalysis is considered 

useful for diagnosis and treatment planning, and furthermore, 

contributes to improved treatment efficacy for IEBAM. 

Disclosures: 

The following authors have nothing to disclose: Nakayuki Naritaka, Mitsuyoshi 

Suzuki, Hajime Takei, Akihiko Kimura, Takao Kurosawa, Takashi Iida, Hiroshi 

Nittono, Toshiaki Shimizu 

1697 

Role of transient elastography to differentiate children 

with biliary atresia from other causes of neonatal 

cholestasis 

Bikrant B. Lal, Rajeev Khanna, Vikrant Sood, Dinesh Rawat, Seema 

Alam; DEPARTMENT OF PEDIATRIC HEPATOLOGY, INSTITUTE 

OF LIVER AND BILIARY SCIENCES, New Delhi, India 

Transient elastography is an important non-invasive tool to 

detect fibrosis. Children with biliary atresia need to be diagnosed 

early as outcome post Kasai surgery depends on age at 

operation. Fibroscan can serve as an important non-invasive 

tool and an alternative to liver biopsy. Objective: To study the 

role of transient elastography to differentiate biliary atresia 

from neonatal hepatitis. To evaluate its efficacy in predicting 

outcome post kasai portoenterostomy. Methodology: All 

patients with neonatal cholestasis were included in the study 

and evaluated as per the instituitional protocol. Transient elastography 

(TE) was done in a 4-hour fasting state using Fibro- 

Scan502, Echosens, Paris, France. Median of 10 successful 

readings was taken and values with IQR>30% or success rate 

of


in experimental and human BA. Methods: Neonatal mice were 

challenged with saline or RRV soon after birth. Gene expression 

was determined by real-time PCR and immune cells were quantified 

by flow cytometry. Histology was performed using H/Estained 

sections. Immunofluorescence was performed using 

antibodies against C5aR, CD68, pan-cytokeratin (CK) and 

Vimentin. Liver wedge biopsies were obtained from patients at 

Kasai and explants at transplantation. Genome-wide expressions 

were obtained from GEO Series: GSE46995. Results: In 

diseased mice, expression levels of Cd68, Cd14 and Mpeg1 

progressively increased from the inception of epithelial injury 

to complete duct atresia (days 3–14, 1.7–4.5 fold above 

saline-injected mice; P=0.001). Proinflammatory macrophage 

polarization correlated with increased expressions of Ccl2, 

Ccl3, Ccl4, Ccr7, Tlr2 and Tlr6 (1.3–8.3 fold above controls; 

P


and 1764 (p=0.04) in the basal core promoter region. In 

patients >12 years (N=75), 3 groups of baseline mutations 

were significantly associated with long-term ALT normalization 

(p


1703 

Dissecting cellular entry mechanisms by enveloped and 

nonenveloped hepatitis E virus 

xin yin, Zongdi Feng; Research Institute at Nationwide Children’s 

Hospital, Columbus, OH 

Hepatitis E virus (HEV) infection is a significant global health 

issue. Recent studies show that while nonenveloped HEV virions 

are shed in feces, the virus circulating in blood is cloaked 

in host membrane and hidden from neutralizing antibodies. 

To better understand HEV infection and pathogenesis, we 

investigate the mechanism by which enveloped HEV (eHEV) 

enters the cell. Gradient purified enveloped and non-enveloped 

HEV (genotype 3, Kernow C1/P6) were inoculated to 

HepG2 cells and cellular pathways and factors involved in 

HEV entry were studied by using pharmacological inhibitors 

and siRNA mediated gene depletion. Compared to non-enveloped 

HEV, cellular attachment of eHEV is much less efficient 

and its entry is also much slower. Entry of eHEV is also more 

sensitive to dynamin-2 and clathrin depletion, suggesting eHEV 

internalization involves clathrin-mediate endocytosis. Knockdown 

of Rab7A and Rab5A had a greater inhibitory effect 

on the entry of eHEV than nonenveloped HEV. In addition, 

treatment of cells with lysosomotropic agents bafilomycin A or 

NH 4 

Cl almost completely blocked the entry of eHEV, but had 

no effect on the entry of nonenveloped HEV, indicating eHEV 

also requires endosomal acidification for its entry. Finally, 

depletion of NPC-1 (Niemann-Pick disease, type C1), a host 

protein involved in cholesterol transport in late endosomes/ 

lysosomes and Ebolavirus entry, greatly reduced entry of eHEV, 

but not nonenveloped HEV. Conclusion: Entry of eHEV involves 

dynamin-2 and clathrin-mediated endocytosis and requires 

endosomal acidification. eHEV envelope is likely degraded in 

endolysosomes in order for the viral capsid to interact with its 

cellular receptor. These results have important implications for 

understanding HEV infection and pathogenesis. 

Disclosures: 

The following authors have nothing to disclose: xin yin, Zongdi Feng 

1704 

Pre-core region mutations analysis helps to delineate 

immune stage in HBeAg+ children with chronic hepatitis 

B 

Ivana Carey, Sanjay Bansal, Sarah Tizzard, Matthew J. Bruce, 

Mary Horner, Kosh Agarwal, Diego Vergani, Giorgina Mieli-Vergani; 

Institute of Liver Studies, Kings College School of Medicine at 

King’s College Hospital, London, United Kingdom 

Interaction between the hepatitis B virus (HBV) and the immune 

system is crucial for the outcome of chronic hepatitis B (CHB). A 

recent study shows that presence of pre-core region mutations 

helps distinguishing between the immune stages of HBeAg+ 

CHB in adults, immunotolerant patients having no mutations 

and higher HBsAg plasma levels. There are no data on pre-core 

region mutations in relation to the immune stage in HBeAg+ 

CHB children. In this retrospective cross-sectional study on 

HBeAg+ CHB children with different alanine-aminotransferase 

(ALT) profiles, we aimed to evaluate the frequency of precore 

region mutations – basal core promoter (BCP) (A1762T/ 

G1764A) and stop codon (A1896G) - and its relationship with 

semi-quantitative HBeAg and HBsAg levels, HBV DNA viral 

load, HBV genotypes and levels of interferon-gamma-inducible 

protein 10 (IP10), a chemokine reflecting the activation status 

of the innate immune system in CHB. Patients: 137 children 

(median age 9.55 years) with HBeAg+ CHB were followed 

for at least 2 years (median 5 years) and, based on their ALT 

profiles, were divided into 3 groups: persistently normal (N) 

ALT (n=64), fluctuating (F) ALT (n=43) and persistently abnormal 

(A) ALT (n=30). Material and methods: Plasma samples at 

diagnosis were used to evaluate BCP and stop codon 1896 

mutations and HBV genotypes by direct population sequencing; 

semi-quantitative HBeAg and HBsAg plasma levels were 

measured by Abbott ARCHITECT® assay [S/CO & log 10 

IU/ 

ml]; HBV DNA viral load by real-time PCR [log 10 

IU/ml] and 

IP10 plasma levels by ELISA [pg/ml]. Results: Frequencies of 

BCP and stop codon mutations were higher in patients with 

fluctuating (F) and abnormal (A) ALT profiles than in patients 

with normal (N) ALT levels (p


Statistical analysis was performed using STATA software 

version 12.0. Statistical significance was defined as P-value 


on established exposure-safety analyses. Other PK parameters 

were similar between the two populations. Conclusion In 

HCV-infected adolescents (12 to< 18 years), SOF 400 mg 

(+RBV) or LDV/SOF 90 mg/400 mg were well tolerated and 

provided comparable plasma exposures to those observed in 

adults. These data confirm the appropriateness of SOF or LDV/ 

SOF adult clinical doses in adolescents. 

Disclosures: 

Brian Kirby - Employment: Gilead Sciences 

Polina German - Employment: GIlead Sciences, Inc; Stock Shareholder: GIlead 

Sciences, Inc 

Bittoo Kanwar - Employment: Gilead Sciences 

Isobel Lakatos - Management Position: Gilead Sciences, INC; Stock Shareholder: 



The following authors have nothing to disclose: Liyun Ni, John Ling 

1708 

HCV infection shifts CD8+ T cell Naïve and Memory 

Subsets in the Maternal Fetal Interface (MFI) 

Melissa A. Sheiko 1 , Rachel McMahan 2 , Lucy Golden-Mason 2 , 

Christine Waasdorp Hurtado 1 , Mona Krull 3 , Silvia Giugliano 2 , 

Angela M. Mitchell 2 , Michael R. Narkewicz 1 , Hugo R. Rosen 2 ; 

1 Pediatrics, Digestive Health Institute, University of Colorado Denver 

School of Medicine, Aurora, CO; 2 Department of Medicine, 

Gastroenterology, University of Colorado Denver School of Medicine, 

Aurora, CO; 3 Ob/Gyn, DenverHealth Medical Center, Denver, 

CO 

Introduction: Women with Hepatitis C (HCV) infection have low 

rates of vertical transmission (VT) to their infants (2-6%) compared 

to HIV and hepatitis B. It has been postulated that this is 

due to an immune response at the maternal fetal interface (MFI). 

The MFI is comprised of the maternal blood, the decidua (the 

part of the placenta that contacts the maternal blood), the placenta, 

and the infant’s blood. Methods: Samples were collected 

from mother and infant pairs with and without HCV. Mononuclear 

cells were isolated from the maternal blood, decidua, 

placenta, and cord blood. Multi-parameter flow cytometry 

was performed, using antibodies to CD3, CD8, CCR7, and 

CD45RA. All cells that were analyzed were CD3+ and CD8+ T 

cells. Naïve cells were also CCR7+CD45RA+, central memory 

cells were CCR7+CD45RA, effector memory cells were CCR7- 

CD45RA-, and terminally-differentiated effector memory cells 

were CCR7-CD45RA+. GraphPad Prism software was used 

for analysis: medians are reported, and p values were determined 

by the Mann Whitney U test. Results: The CD8+ T cells 

from the decidua showed differences in all subsets of CD8+ 

T cells between controls and HCV-exposed subjects: there 

were fewer naïve cells in HCV-exposed (15.7% versus 36.0%; 

p


for inflammatory bowel disease (250-450). Our study did not 

reveal specific goal levels to aim for in the treatment of AIH, but 

would suggest that patients should be maintained at the lowest 

level that keeps them in remission. 

of PSC and 64% of Overlap. Conclusion: The nomenclature of 

autoimmune liver disease remains inadequate and in need of 

consensus. Of young adult PSC patients, half presented with an 

AISC phenotype, and of young adult AIH-PSC overlap, nearly 

half presented as AIH. Evaluating whether immunosuppressive 

therapy in childhood prevents ultimate disease progression 

remains an important goal for prospective studies. 

Characteristics of children with AIH, PSC and AISC at diagnosis 

Disclosures: 

The following authors have nothing to disclose: Melissa A. Sheiko, Kelley E. 

Capocelli, Shikha Sundaram, Zhaoxing Pan, Annette McCoy, Cara L. Mack 

Disclosures: 







Deirdre A. Kelly - Consulting: Sanofi Pasteur; Grant/Research Support: BMS, 

Astellas, Acitllion, MSD, Roche 




The following authors have nothing to disclose: Jeremy K. Rajanayagam, Ye H. 

Oo 

1710 

Long-term follow up of childhood autoimmune liver disease: 

the importance of resolving nomenclature 

Jeremy K. Rajanayagam 1,3 , James W. Ferguson 2 , Ye H. Oo 2,3 , 

David H. Adams 2,3 , Deirdre A. Kelly 1 , Gideon Hirschfield 2,3 ; 1 Liver 

Unit, BIrmingham Children’s Hospital, Birmingham, United Kingdom; 

2 Liver Unit, Queen Elizabeth Hospital, Birmingham, United 

Kingdom; 3 Centre for Liver Research, University of Birmingham, 

Birmingham, United Kingdom 

Background: The nomenclature and long term outcomes for 

patients with autoimmune liver diseases transitioned to adult 

practice lacks consensus. Aim: To study the nature and progression 

of childhood onset autoimmune liver disease in patients 

transitioned from pediatric to adult liver care. Methods: Clinical 

review of transitioned patients with standard definitions 

of AIH and PSC was performed. AISC was defined as cholangiographic 

and/or histologic evidence of biliary involvement, 

clinical features of autoimmunity (increased IgG, and positive 

autoantibodies), and histological findings of AIH. The term AIH- 

PSC overlap was used for concomitant occurrence of clinical, 

biochemical, serological and/or histological features of PSC 

and AIH. Results: 73 children with pediatric autoimmune liver 

disease were transitioned over a median follow-up of 13 years 

(IQR 8-16 years). At presentation, 52 were diagnosed with AIH 

(38F), 4 with PSC (1F); 17 with AISC (8F) (Table). IBD occurred 

in 4% of AIH, 25% of PSC and 65% of AISC at diagnosis with 

liver disease. Median IAIHG score at diagnosis was 19 (IQR 

17-21), 4 (IQR 3.5-5) and 12 (IQR 10-14) for AIH, PSC and 

AISC, respectively. Children with AIH & AISC were treated 

with corticosteroids and azathioprine, with ursodeoxycholic 

acid added in AISC. 10/52 (19%) patients initially presenting 

as AIH had phenotypic progression by last adult review: 6 

(11.5%) developed an overlap syndrome whilst 4 (7.6%) had 

clinically dominant PSC. 8/17 (47%) AISC patients retained 

an adult overlap phenotype, but 9 (53%) had clinically dominant 

PSC. Those presenting as childhood PSC (n=4) remained 

phenotypically as PSC in adulthood. Thus by last review in 

adult care the clinical diagnoses were AIH n=42, PSC n=17 

and Overlap AIH/PSC n=14. IBD occurred in 5% of AIH, 65% 

1711 

Autoimmune Liver disease in Children with Sickle Cell 

Disease 

Suttiruk Jitraruch 2 , Emer Fitzpatrick 2 , Maesha Deheragoda 2 , Giorgina 

Mieli-Vergani 2 , Sue Height 1 , Nedim Hadzic 2 , Marianne 

Samyn 2 ; 1 Paediatric Haematology, King’s College Hospital, London, 

United Kingdom; 2 Institute of Liver Studies, King’s College 

Hospital, London, United Kingdom 

Sickle cell disease (SCD) is an autosomal recessive haemoglobinopathy 

resulting in intermittent haemolysis and microvascular 

occlusions. Hepatic involvement varies from asymptomatic gallstones 

to life-threatening acute liver failure (ALF) and cirrhosis. 

Aim: To characterise the clinical features, natural history and 

outcome of autoimmune liver disease (AILD) in patients with 

SCD. Methods: We performed a retrospective review of SCD 

patients with hepatic dysfunction referred to our centre from 

1999-2015. The demographic, clinical, laboratory, histological 

and radiological features, management and outcome were 

studied. The diagnostic criteria included: positive serum autoantibodies, 

hypergammaglobulinemia, compatible histology 

and absence of viral/metabolic causes. Results: Of 83 SCD 

patients with hepatic dysfunction, 13 (16%), (10 female) were 

diagnosed with AILD at a median age of 11 (range, 3.4-16) 

years. Eleven were homozygote for HBSS and 2 required regular 

transfusions. Family history for autoimmune disease was 

positive in 2. Two patients presented with ALF (INR 2.7 and 

1.8). In two patients parvo B19 induced aplastic crisis preceded 

AILD diagnosis, 2 and 6 months, respectively. At presentation, 

anti-nuclear and anti-smooth muscle autoantibodies 

were in range 1/20-1/2560 and 1/10-1/320, respectively, 

median AST 294 (range, 67-814) U/L, IgG 33.5 (range, 13.7- 

43.7) g/L and INR 1.32 (range, 1.01-2.7). Ultrasonography 

showed enlarged lymph nodes at porta/superior to pancreas 

in 4, gall stones in 3, and splenomegaly in 5 patients. On 

MRCP five children had radiological features of cholangiopathy; 

4 at presentation and 1 three years later. Liver biopsy 

was performed in 11 (6 via transjugular route) without complications; 

9 showed interface hepatitis without cholangiopathy,


one obtained after treatment and another one was inadequate. 

Treatment included ursodeoxycholic acid (12), prednisolone 

(12), azathioprine (8) and mycophenolate mofetil (1). After a 

median follow up of 3.8 (range, 0.2-14.3) years, 10 patients 

are alive with 2 lost to follow up. One patient died following 

intracranial haemorrhage. One patient required liver transplantation, 

6.4 years after diagnosis due to recurrent biliary 

sepsis. Four patients were in full and five in partial remission. 

Four patients (2 males) were diagnosed with ulcerative colitis 

2 before and 2 after AILD. Conclusion: AILD is not uncommon 

in patients with SCD with a strong female preponderance. It 

responds well to standard treatment. Liver biopsy can be helpful 

to confirm the diagnosis. Ulcerative colitis was more common 

in the boys and should be excluded. 

Disclosures: 

Nedim Hadzic - Consulting: Alnylam, Cambridge, MA; Speaking and Teaching: 

Synegeva UK 

The following authors have nothing to disclose: Suttiruk Jitraruch, Emer Fitzpatrick, 

Maesha Deheragoda, Giorgina Mieli-Vergani, Sue Height, Marianne 

Samyn 

1712 

“Hepatic Granulomas: 18 Year Experience of a Tertiary 

Paediatric Liver Centre in the UK” 

Marumbo P. Mtegha, Katie Hunt, Nedim Hadzic, Maesha Deheragoda, 

Sanjay Bansal; Department of Paediatric Hepatology, 

Gastroenterology and Nutrition, King’s College Hospital, London, 


Objective Hepatic granulomas are reported in 2-15% of all 

liver biopsies in adult studies . Up to 30% have no identifiable 

etiology. The incidence in the paediatric population is not well 

established. We describe the prevalence, character and aetiology 

of this pathology at our centre. Methods Retrospective 

analysis of all liver biopsies showing granulomas and their 

clinical data between 1996-2014. Results Granulomas were 

identified in 44 (0.32%) of 13542 paediatric liver biopsies 

perfomed. Native Liver 31 (70.4%) patients had granuloma 

within their native liver (Male 52%). The median age at presentation 

was 3.7yrs (range 0.3–11.8). The commonest presentation 

was abnormal liver function tests (LFTs) with or without 

cholestasis. 4 patients presented with isolated or a combination 

of hepatomegaly, splenomegaly and lymphadenopathy. 

Etiologies are listed in Table 1. Histologically, non-caseating 

and epithelioid granulomas were found in 26 cases. Lipogranuloma 

were described in 2 cases, one each in Non-alcoholic 

fatty liver disease and HIV infection. There were 3 cases of 

caseating granuloma (2 Sarcoidosis and Tuberculosis). Transplanted 

Liver 13 (29.6%) patients had granulomas within their 

allograft. The median age at liver transplant was 1.4 years 

(range 0.3-15.9). Median time from transplant to granuloma 

detection was 0.2yrs (range 8 days to-8.1yrs). All granulomas 

were non-caseating and epithelioid. Acute cellular rejection 

was noted in 3 cases, chronic rejection in 2. 7 cases showed 

non-specific lobular hepatitic changes, the last showed early 

erythrophagocytosis. Conclusions Hepatic granulomas in children 

are rare with idiopathic neonatal hepatitis being the most 

common condition in the native liver. In the post transplant 

setting, they are more common in the younger age group and 

in the first few months after transplant. In this setting, they may 

be associated with acute or chronic rejection. 

*= Persistent biochemical or radiological abnormalities 

**=Normalization of biochemistry and radiology 

***=Incomplete follow-up data 

Disclosures: 

Nedim Hadzic - Consulting: Alnylam, Cambridge, MA; Speaking and Teaching: 

Synegeva UK 

The following authors have nothing to disclose: Marumbo P. Mtegha, Katie Hunt, 

Maesha Deheragoda, Sanjay Bansal 

1713 

Differentiating autoimmune hepatitis from lupus-associated 

hepatitis: A paradigm shift in approach to elevated 

liver enzymes 

Andreanne Benidir 1 , Brian M. Feldman 3 , Anna Goldenberg 2 , Earl 

Silverman 3 , Sindhu Johnson 4 , Binita M. Kamath 1 ; 1 Gastroenterology, 

Hepatology and Nutrition, The Hospital for Sick Chidlren, 

Toronto, ON, Canada; 2 Computer Science, Research Institute - The 

Hospital for Sick Children, Toronto, ON, Canada; 3 Rheumatology, 

The Hospital for Sick Children, Toronto, ON, Canada; 4 Rheumatology, 

Toronto Western Hospital, Toronto, ON, Canada 

Background:Whether autoimmune hepatitis(AIH), originally 

“lupoid hepatitis”, and lupus(SLE)-associated hepatitis correspond 

to two distinct entities or are part of the same disease 

spectrum is controversial . Both are diagnosed based on 

expert-consensus and complex constellations of clinical, serological 

and histological features. There is the potential for an 

inherent flaw in defining these diseases based on human-driven 

criteria. We performed a rigorous hypothesis-generating study 

that removes patients’ current diagnostic labels and clusters 

them based on their broad clinical data. Aim:To determine if 

distinct disease entities can be objectively determined from a 

heterogeneous group of children with non-infectious, -toxic, 

-genetic causes of elevated liver enzymes. Methods:A retrospective 

cohort of pediatric patients(age


grated clusters with diagnosis-based groups was 0.44(perfect 

similarity =1). Within the 8 groups, children with PSC were 

divided into 2. Those with AIH clustered across multiple groups 

that mostly did not include lupus patients. Conclusion:Our data 

suggests AIH and SLE-associated hepatitis exist as 2 distinct 

conditions that cluster differently with other causes of elevated 

liver enzymes. Identifying key differentiating groups of features 

that drive the similarities between children with elevated liver 

enzymes may assist in better future management and understanding 

of their disease. 

Disclosures: 

The following authors have nothing to disclose: Andreanne Benidir, Brian M. 

Feldman, Anna Goldenberg, Earl Silverman, Sindhu Johnson, Binita M. Kamath 

1714 

Screening for Mitochondrial Disease in Pediatric Acute 

Liver Failure: The Role of Lactate and Pyruvate 

Amy G. Feldman 1,5 , Ronald J. Sokol 1,5 , Regina M. Hardison 4 , 

Estella M. Alonso 2,6 , Robert H. Squires 3,7 , Michael R. Narkewicz 

1,5 ; 1 Pediatric Gastroenterology, Hepatology and Nutrition, 

Children’s Hospital Colorado, Aurora, CO; 2 Department of Pediatrics, 

Ann and Robert H Lurie Children’s Hospital of Chicago, 

Chicago, IL; 3 Division of Gastroenterology, Children’s Hospital 

of Pittsburgh, Pittsburgh, PA; 4 Graduate School of Public Health, 

University of Pittsburgh, Pittsburgh, PA; 5 University of Colorado 

School of Medicine, Aurora, CO; 6 Northwestern University Feinberg 

School of Medicine, Chicago, IL; 7 University of Pittsburgh 

School of Medicine, Pittsburgh, PA 

Objectives: Mitochondrial disorders can cause pediatric acute 

liver failure (PALF). Screening for mitochondrial disorders using 

serum lactate and lactate/pyruvate (L:P) molar ratio has been 

recommended, yet the diagnostic utility of these tests in PALF 

has not been systemically analyzed. The goals of this study 

were to determine whether 1) the L:P ratio is a sensitive and 

specific test to screen for a mitochondrial, respiratory chain, or 

fatty acid oxidation (Mito) disorder in children with PALF and 

(2) the L:P ratio correlates with clinical outcome at 21 days. 

Study Design: A retrospective review was conducted of demographic, 

clinical, laboratory, and outcome data for patients in 

the NIH-supported PALF Study dataset who had lactate and 

pyruvate levels collected on the same day (within 7 days of 

enrollment) and a serum lactate level ≥2.5 mmol/L. The Kruskal-Wallis 

test and Pearson chi-square were used to compare 

continuous variables and proportions, respectively, among the 

diagnosis groups. Spearman correlations were used to estimate 

the association between L:P and clinical labs. Multinomial 

logistic regression was used to estimate the effect of L:P group 

on outcome. Results: Of 986 subjects, 110 had lactate and 

pyruvate levels collected on the same day, 74 (67 %) of whom 

had a lactate level ≥2.5 mmol/L. Of these 74 participants, 

39 (53%) were male; median age was 2.0 years (IQR 0.36- 

5.9). Six (8%) had a Mito disorder, 22 (29.7%) had another 

confirmed diagnosis, and 46 (62.2%) had an indeterminate 

diagnosis. There were no significant differences in the lactate 

levels, pyruvate levels, or L:P ratios between these three groups. 

Although 6 Mito group subjects had lactate ≥2.5mmol/L, only 

2 had L:P ratio ≥25. The percent with elevated L:P ratio≥ 25 did 

not differ by diagnostic group (33% Mito, 64% other diagnosis 

and 54% indeterminate diagnosis). There was no significant 

relationship between the L:P ratio and the AST/ALT ratio, INR, 

ammonia, creatinine kinase, acylcarnitine profile, or glucose 

levels nor was there a significant relationship between initial 

L:P ratio and clinical outcomes at 21 days (alive with native 

liver, alive with transplanted liver, dead). Conclusions: A high 

serum lactate and/or elevated L:P ratio were common in all 

causes of PALF, were not diagnostic for a mito cause, and did 

not predict 21 day clinical outcome. This suggests that either 

mito dysfunction is common in all causes of PALF, that mito 

disorders are misdiagnosed, or that L:P ratio is not specific for 

mito disorders. Patient age and clinical features remain the 

most helpful diagnostic clues for mito disorders, which must be 

confirmed by genetotyping or tissue enzymology. 

Disclosures: 




Michael R. Narkewicz - Consulting: Vertex; Grant/Research Support: Novartis, 

Vertex; Stock Shareholder: Merck 

The following authors have nothing to disclose: Amy G. Feldman, Regina M. 

Hardison, Estella M. Alonso, Robert H. Squires 

1715 

Nocturnal Hypoxia is associated with Hedgehog Signaling 

and Severity of Pediatric Non-Alcoholic Fatty Liver 

Disease (NAFLD) 

Shikha Sundaram 1 , Marzena Swiderska-Syn 2 , Ronald J. Sokol 1 , 

Zhaoxing Pan 1 , Ann C. Halbower 1 , Kelley E. Capocelli 1 , Kristen 

N. Robbins 1 , Anna Mae Diehl 2 ; 1 Children’s Hospital Colorado, 

Aurora, CO; 2 Duke University, Durham, NC 

Chronic intermittent nocturnal hypoxia is associated with 

NAFLD progression. Deregulation of the Hedgehog (Hh) pathway 

has also been implicated in NAFLD pathogenesis and 

progression. Objective: To determine relationships between 

obstructive sleep apnea (OSA), hypoxia, and Hh signaling in 

pediatric NAFLD. Methods: 31 adolescents with biopsy-proven 

NAFLD (mean age: 13 ± 1.9 yrs; mean BMI z score: 2.2 ± 

0.3, 65% male, 87% Hispanic) underwent polysomnogram, 

liver histology scoring, and laboratory testing. Sonic hedgehog 

(SHh), Indian Hh (IHh), Glioblastoma associated oncogene 

2 (Gli2), and markers of progenitors (keratin 7- K7) and 

myofibroblasts (α-SMA) were evaluated by immunohistochemistry. 

Results: Subjects with (68%) and without (32%) OSA/ 

hypoxia had similar aminotransferases, serum lipids, inflammatory 

and insulin resistance markers. As expected, α-SMA 

generally correlated with fibrosis stage (r=0.44, p=0.02). Steatosis 

was inversely correlated with IHh (r=-0.48, p=0.009) 

and the NAS score correlated with SHh (r=0.39, p=0.03) and 

α- SMA (r=0.42, p=0.02). SHh correlated with AST (r=0.64, 

p=0.0001) and ALT (r=0.51, p=0.003). Gli2 also correlated 

with AST (r=0.47, p=0.007) and ALT (r=0.43, p=0.02). AST 

correlated with K7 (r=0.45, p=0.01), and ALT with α-SMA 

(r=0.51, p=0.005). Subjects with OSA/hypoxia had higher 

mean Apnea Hypopnea Index (AHI), % time oxygen saturation 

(SaO 2 

)


and relates to NAFLD disease severity. We speculate that tissue 

hypoxia allows for functional activation of Hypoxia Inducible 

Factor 1α with subsequent induction of genes important in 

epithelial-mesenchymal transition, including SHh, and NAFLD 

progression. 

Disclosures: 




The following authors have nothing to disclose: Shikha Sundaram, Marzena 

Swiderska-Syn, Zhaoxing Pan, Ann C. Halbower, Kelley E. Capocelli, Kristen N. 

Robbins, Anna Mae Diehl 

1716 

Risk Factors Associated with Cystic Fibrosis-associated 

Liver Disease (CFLD) 

Tamir A. Miloh 1,2 , Amanda Pope 1 , John Daye 3 , Chengcheng Hu 3 , 

Peggy Radford 1 ; 1 Phoenix Children, Phoenix, AZ; 2 Mayo Clinic, 

Scottsdale, AZ; 3 University of Arizona, Tucson, AZ 

Cystic fibrosis-associated liver disease (CFLD) affects approximately 

30% of patients and is a significant cause of morbidity 

and mortality. Aims: Analyze data from the CF registry to 

determine the predictors significant towards the development 

and progression of CFLD. We analyzed 39642 unique and 

valid observations documented on routine annual visits from 

1986 to present. Categorical, quantitative and ordinal variables 

were analyzed using Pearson’s Chi-squared test, 2-group 

t-test with Welsh degree of freedom modification and the 

independent 2-group Mann-Whitney U-test, respectively. The 

following phenotypes were further analyzed (n and %): any 

liver disease (11,002, 28%), gallstones (480, 1.7%), cirrhosis 

(1735, 4.5%), steatosis (150, 0.5%), non-cirrhotic liver disease 

(3513, 9%) and other (5124, 13.2%). Complications of cirrhosis 

(n and % of patients with cirrhosis) include gastric or esophageal 

varices (191, 22.9%), gastrointestinal bleeding (292, 

22.3%), splenomegaly and hypersplenism (302, 36%) and 

ascites (82, 9.8%). Variables were statistically significant at 

the p-valueG (54) and 3876delA (8.4). 

The following reduced risk of CFLD: Caucasian (0.65) and 

diagnosis on newborn screening (NS) (0.4). (b) Gallstones: FTT 

(1.43), steatorrhea (1.73), respiratory infection with pseudomonas 

(2.79), burkholderia cepacia (1.7), fungal (1.84). Protective: 

male gender (0.6) and NS (0.17). (c) Cirrhosis: male 

gender (1.4), FTT (1.43), meconium ileus (1.75), steatorrhea 

(1.33), respiratory infection with staphylococcal (1.4), pseudomonas 

(3.35), burkholderia cepacia (1.65), fungal (1.7) and 

F508 mutations (1.44). Mean chloride sweat concentration 

was 5.62 mmol/L higher than those without cirrhosis. Protective: 

NS (0.28) and mutations: R117H (0.23), 2789+5G>A 

(0.05) and 3849+10kbC>T (0.16). (d) Steatosis: mutations 

Q890X (26.66) and 218delA (8.86). The mean age of death 

for patients with cirrhosis was 2.24 years less than those without 

cirrhosis. Conclusion: Analyzing the largest CF database 

revealed that CFLD leads to earlier mortality and male gender, 

non-Caucasian, meconium ileus, malnutrition, increased sweat 

chloride, certain respiratory infections and mutation were associated 

with increased risk of CFLD and other mutations were 

protective. 

Disclosures: 

The following authors have nothing to disclose: Tamir A. Miloh, Amanda Pope, 

John Daye, Chengcheng Hu, Peggy Radford 

1717 

Hepatic Manifestations in Adenosine Deaminase-Deficient 

Severe Combined Immune Deficiency 

Shilpa Lingala 1 , Elizabeth Garabedian 2 , Fabio Candotti 2 , David 

E. Kleiner 4 , Kenneth J. Wilkins 3 , Theo Heller 1 , Christopher Koh 1 ; 

1 Liver Diseases Branch, National Institute of Diabetes and Digestive 

and Kidney Diseases, NIH, Bethesda, MD; 2 National Human 

Genome Research Institute, NIH, Bethesda, MD; 3 Office of the 

Director, National Institute of Diabetes, Digestive and Kidney Diseases, 

Bethesda, MD; 4 Laboratory of Pathology, National Cancer 

Institute, Bethesda, MD 

Background: Adenosine deaminase-deficient severe combined 

immune deficiency (ADA-SCID) is a rare autosomal recessive 

metabolic disorder that occurs in


1718 

Autosomal Recessive Polycystic Kidney Disease: Not just 

a Renal Disease 

Sarika Rohatgi 1 , William E Sweeney 2 , Emma Schwasinger 2 , Nicholas 

Kampa 2 , Ellis D. Avner 2 ; 1 Pediatric Gastroenterology, Medical 

college of Wisconsin, Milwaukee, WI; 2 Pediatric Nephrology, 

Medical college of Wisconsin, Wauwatosa, WI 

Introduction: Autosomal Recessive Polycystic Kidney Disease/ 

Congenital Hepatic Fibrosis (ARPKD/CHF) is an inherited hepatorenal 

fibrocystic disease with a wide spectrum of dual organ 

developmental abnormalities, secondary to PKHD1 mutations. 

Renal pathophysiology of ARPKD/CHF is well understood and 

clinical trials targeting renal disease are near. CHF is not well 

studied and warrants investigation due independent disease 

progression in the two organs. We hypothesize that proliferation 

and fibrosis are two separate but overlapping processes 

and identification of the ‘Critical stage’ of disease is important 

to achieve goal therapeutic results in CHF. Aim and Methods: 

PCK rat, an orthologous model with PKHD1 mutation, consistently 

develops dual organ abnormalities as in human disease, 

providing the best model for investigation. We performed a 

systematic characterization of morphological changes in development 

and progression of CHF in PCK Rats, correlating them 

with biliary ductal ectasia, proliferation, apoptosis and periportal 

fibrosis at progressive stages of disease. Our goal was to 

identify ‘critical stages’ of the disease to develop liver specific 

therapies. Five male rats each at 0, 30, 60, 90, 120 and 

150 days of age were evaluated and physiologic data (body, 

liver, kidney, intestine and spleen weights) collected. Serial 

sections of these livers were stained with Masson’s Trichrome 

stain to score fibrosis and immunohistochemistry (IHC) was 

done to identify cholangiocytes, proliferation and apoptosis. 

Morphometric analysis was done using ImageJ to quantitate 

disease progression. Results: Liver weight to body weight ratios 

increased as disease progressed. Morphometric analyses of 

liver tissues show consistent enlargement and deformation of 

biliary ducts with increasing age. A 50% increase in cyst numbers 

and cyst area per surface area of the liver with increased 

proliferation of cholangiocytes and mesenchymal cells was 

observed between ages of 30 and 90 days. Thereafter, proliferation 

gradually decreased to negligible by day 150. Periportal 

fibrosis index increased to 39% at day 90 and to 47% 

at 150 days. Fibrosis score increased from 0 to 2 between 

30 and 90 days and to 4 at 150 days. Fibrosis was more 

prominent in hilum around larger cysts. Apoptosis increased 

steadily with age. Conclusion: Rapid increase in proliferation 

and fibrosis at day 90 and decrease thereafter suggest that 

‘critical stage’ of the disease is before 90 days. Given the wide 

variability in rate of disease progression in CHF, therapies 

aimed at reducing proliferation will require a histopathological 

assessment of the liver disease to provide maximum benefit. 

Disclosures: 

The following authors have nothing to disclose: Sarika Rohatgi, William E Sweeney, 

Emma Schwasinger, Nicholas Kampa, Ellis D. Avner 

1719 

Recurrent recessive mutation in DGUOK causes non-cirrhotic 

intrahepatic portal hypertension in the absence of 

liver synthetic dysfunction 

Silvia Vilarinho 4,1 , Sinan Sari 2 , Guldal Yilmaz 3 , Buket Dalgic 2 , 

Richard P. Lifton 1,5 ; 1 Department of Genetics, Yale University 

School of Medicine, New Haven, CT; 2 Department of Pediatrics, 

Gazi University - Faculty of Medicine, Ankara, Turkey; 3 Department 

of Pathology, Gazi University - Faculty of Medicine, Ankara, 

Turkey; 4 Department of Internal Medicine, Section of Digestive 

Diseases, Yale University School of Medicine, New Haven, CT; 

5 Department of Internal Medicine, Yale University School of Medicine, 

New Haven, CT 

Liver cirrhosis is the most frequent cause of portal hypertension, 

but a variety of other diseases may cause elevated pressure 

in the portal venous system. However, when all known causes 

of portal hypertension have been ruled out, the diagnosis of 

idiopathic non-cirrhotic intrahepatic portal hypertension is 

established. In the literature, the terminology of this medical 

condition is ambiguous and several other terms such as hepatoportal 

sclerosis, obliterative venopathy, idiopathic portal 

hypertension, incomplete septal cirrhosis and nodular regenerative 

hyperplasia are used interchangeably. Despite progress 

in the diagnosis and management of this clinical entity, 

its etiopathogenesis remains elusive. Interestingly, idiopathic 

non-cirrhotic intrahepatic portal hypertension has also been 

reported in infancy and childhood, occasionally affecting more 

than one family member, suggesting that an unrecognized 

Mendelian trait may underpin the onset and development of 

portal hypertension in a subset of these subjects. To investigate 

this hypothesis, we applied unbiased whole-exome capture 

and DNA sequencing, in contrast to a hypothesis-driven 

approach, to nine Turkish subjects with onset of intrahepatic 

portal hypertension of indeterminate etiology during infancy 

or early childhood. Whole-exome sequencing allows us to 

determine the existence of recurrent mutations across samples 

and to assess the burden of rare variants in individual gene(s) 

greater than expected by chance. We found three individuals, 

in two unrelated families, that shared an identical rare 

homozygous mutation in deoxyguanosine kinase, encoded by 

the DGUOK nuclear gene, at a highly conserved amino acid 

position (p.Asn46Ser). The probability of finding two unrelated 

instances of this identical homozygous variant in DGUOK by 

chance is conservatively estimated to be 5.2x10 -12 , providing 

extremely strong statistical support for the role of this mutation 

in non-cirrhotic intrahepatic portal hypertension. DGUOK is an 

enzyme responsible for phosphorylation of purine deoxyribonucleosides 

in the mitochondrial matrix and its dysfunction has 

been previously associated with mitochondrial depletion syndrome 

type 3. In contrast to this syndrome, none of our index 

cases demonstrated signs of hepatic synthetic dysfunction for 

a follow-up period of 6 to 17 years. In conclusion, our study 

provides a new insight into the mechanisms mediating non-cirrhotic 

intrahepatic portal hypertension, defines a previously 

unrecognized form resulting from recessive N46S mutation in 

DGUOK, and expands the phenotypic spectrum of DGUOK 

deficiency beyond our current understanding. 

Disclosures: 

The following authors have nothing to disclose: Silvia Vilarinho, Sinan Sari, 

Guldal Yilmaz, Buket Dalgic, Richard P. Lifton


1720 

Liver Involvement in Turner’s Syndrome, Is Associated 

with Extra Hepatic Disease Manifestations 

Ohad Etzion 1 , Theo Heller 1 , Ma Ai Thanda Han 1 , Ruchi Patel 1 , 

Christopher Koh 1 , David E. Kleiner 1,2 , Omar Mousa 1 , Rohit 

Loomba 1 , Vladimir Bakalov 2 , Carolyn Bondy 2 ; 1 Liver Disease 

Branch, NIH, Rockville, MD; 2 Heritable Disorders Branch, NICHID- 

NIH, Bethesda, MD 

Introduction: Turner’s syndrome (TS) is a genetic disorder 

caused by complete, partial lack or structural abnormalities 

of the X chromosome. Liver abnormalities are commonly 

described in patients with TS, however, little is known about 

its association with other clinical features of this syndrome. 

Aim: To characterize the spectrum of hepatic involvement and 

its correlation with non-invasive biomarkers and systemic manifestations 

of disease, in a cohort of patient with TS. Methods: 

From 2004-2009, patients with TS followed at the NIH Clinical 

Center, were evaluated for suspected liver disease. Biochemical 

tests, imaging studies and liver biopsy were performed 

as clinically indicated, and assessed for its association with 

documented extra hepatic disease manifestations. Results: 133 

patients (median age 43 years) were evaluated during this 

period. Abnormal ALT, AST and ALP values were found in 47%, 

40% and 21% of subjects respectively. Sonographic signs of 

steatosis were evident in 42% of patients. Echocardiography 

detected congenital heart defects in 42%, major venous anomalies 

in 13.4%, and aortic or major arterial anomalies in 8.9% 

of subjects. Elevated ALT was associated with presence of 

major venous anomalies (p=0.01), but not with arterial vascular 

anomalies or congenital heart defects. Association between 

hepatic steatosis on ultrasound and major venous anomalies 

trended towards statistical significance (p=0.053). Twenty-four 

patients underwent liver biopsy. Only 1 biopsy was staged as 

Ishak 3, with the rest showing Ishak scores of 0 or 1. Steatosis 

was diagnosed in 11 (46%) cases, and nodular regenerative 

hyperplasia (NRH) in 6 (25%). Portal tracts with missing bile 

ducts, arteries or veins were observed in 78%, 69% and 65% 

of biopsies respectively. Presence of a major artery anomaly 

was associated with higher rate of missing veins in the portal 

tracts (p=0.02), but not with bile duct or artery dropout. 

Congenital heart defects, major venous anomalies and other 

clinical features such as webbed neck and horseshoe kidney 

were associated with neither liver enzyme abnormalities, nor 

with liver histological findings. Conclusion: Liver enzyme abnormalities 

are common in TS and may reflect unique patterns of 

liver injury. As previously reported, steatosis is a dominant form 

of liver injury in TS. Abnormalities in hepatic microvasculature 

are a common finding in TS that are associated with systemic 

vascular anomalies. These findings suggest that liver vascular 

changes in TS represent an organ specific manifestation of a 

systemic disorder of vascular development, which evolves from 

the underlying chromosomal abnormality. 

Disclosures: 






The following authors have nothing to disclose: Ohad Etzion, Theo Heller, Ma 

Ai Thanda Han, Ruchi Patel, Christopher Koh, David E. Kleiner, Omar Mousa, 

Vladimir Bakalov, Carolyn Bondy 

1721 

NOTCH2 variants in cholestatic liver disease 

Tassos Grammatikopoulos 1 , S. Strautnieks 2 , Melissa Sambrotta 1 , 

Pierre Foskett 2 , Maesha Deheragoda 2 , A. S. Knisely 2 , Richard J. 

Thompson 1 ; 1 Institute of Liver Studies, Division of Transplantation 

Immunology and Mucosal Biology, King’s College London, London, 

United Kingdom; 2 Institute of Liver Studies, King’s College 

London, London, United Kingdom 

Background: Alagille syndrome (AGS) is an autosomal dominant 

multisystem disorder associated with mutations in JAG1 

and NOTCH2 in respectively 94% and ~1% of affected 

patients. Aim: We report eight cholestatic patients with novel, 

or rare NOTCH2 variants. Methods: After biliary atresia and 

α-1-antitrypsin deficiency were ruled out, cholestatic patients 

in whom clinical features and/or liver histology findings could 

not exclude AGS were assessed. NOTCH2 screening was 

performed as part of diagnostic next generation sequencing. 

Results: Among 302 patients sequenced using a custom-designed 

cholestasis gene panel, heterozygous variants in 

NOTCH2 were identified in 8 (5 male) patients. Seven variants 

were missense changes. One was protein-truncating (Table). 

Median age at presentation was 5 weeks (range, 2wks-30yrs) 

with jaundice (7), hepatosplenomegaly (1), failure to thrive (4), 

pale stools (5), raised transaminases (7) and pruritus (2). Vertebral 

and ophthalmological and cardiac abnormalities were not 

found in screened patients. Facial characteristics typical of AGS 

were identified in patient 1 and in his maternal grandmother 

who had the same genetic variant. Liver microscopy showed 

lobular cholestasis (5), bile duct paucity (2), and fibrosis (3); 

cytokeratin 7, expressed in attenuated biliary radicles, was 

aberrantly expressed in hepatocytes (2). Abdominal sonography 

and magnetic resonance cholangiography showed bile 

duct irregularities (2), abnormal gallbladder (1) and renal cysts 

(1). Median serum bilirubin was 105 μmol/L [5-227], AST 

86 IU/L [31-319], ALT 90 IU/L [33-195], GGT 172 IU/L [27- 

389], albumin 38 g/L [35-30], creatinine 43 μmol/L [13-89], 

INR 0.98 [0.94-1.1] and cholesterol 3.89 mmol/L [1.9-5.1]. 

Patients 2 and 6 underwent liver transplantation before their 

1 st birthdays. Family histories included jaundice and gallstones 

in maternal grandparents in patient 3, heart murmur in mother 

of patient 2 and similar findings on liver biopsy in father of 

patient 6. Conclusion: We report the identification of eight 

probands with cholestasis and variants in NOTCH2. We could 

find clinical features associated with AGS in only a minority of 

these patients. None met diagnostic criteria for AGS. 

Disclosures: 

Richard J. Thompson - Grant/Research Support: Shire; Speaking and Teaching: 

Shire 

The following authors have nothing to disclose: Tassos Grammatikopoulos, 

S. Strautnieks, Melissa Sambrotta, Pierre Foskett, Maesha Deheragoda, A. S. 

Knisely


1722 

Liver Disease in Subjects with N-Glycanse-1 Deficiency 

Shilpa Lingala 1 , David E. Kleiner 6 , Christina Lam 2 , Lynne A. 

Wolfe 3,5 , Carlos R. Ferreira 2 , Donna Krasnewich 4 , William A. 

Gahl 2,5 , Marc G. Ghany 1 ; 1 Liver Diseases Branch, National Institute 

of Diabetes and Digestive and Kidney Diseases, National 

Institutes of Health, Bethesda, MD, Betheda, MD; 2 Medical Genetics 

Branch, National Human Genome Research Institute, NIH, 

Bethesda, MD; 3 Undiagnosed Diseases Program, NIH, Bethesda, 

MD; 4 Division of Genetics and Developmental Biology,, National 

Institute of General Medical Sciences, NIH, Bethesda, MD; 5 Office 

of the Clinical Director, National Human Genome Research Institute, 

NIH, Bethesda, MD; 6 Laboratory of Pathology, National Cancer 

Institute, Bethesda, MD 

Background: N-glycanase 1 (NGLY1) deficiency is a newly 

described inherited disorder of glycoprotein degradation. 

N-glycanase is a cytosolic enzyme that cleaves intact glycans 

off of misfolded N-linked glycoproteins after they have been 

processed by the endoplasmic reticulum associated degradation 

pathway. Deficiency of NGLY1 in humans manifests as 

developmental delay, multifocal epilepsy, involuntary movements, 

low tear production and elevation of liver-associated 

enzymes. Aim: To describe the spectrum of liver disease in 

subjects with NGLY1 deficiency. Methods: 11of 27 subjects 

with confirmed NGLY1 deficiency worldwide were evaluated 

at the Clinical Center, NIH between June 2014-May 2015. 

Some have been previously reported. A careful history and 

physical exam were performed and all available retrospective 

clinical data including liver biopsy were reviewed. At the Clinical 

Center, subjects underwent a comprehensive evaluation 

for liver disease including liver-associated enzymes, serologic 

testing for known causes of chronic liver disease, abdominal 

ultrasound and transient elastography to assess liver stiffness. 

Liver biopsies, if available, were evaluated by an expert hepatopathologist. 

Results: There were 6 females and 5 males with 

a mean age 9.9 years (2.4-21 yrs.). All were Caucasian. 

6/11(55%) had a history of neonatal jaundice, 50% of whom 

required phototherapy. 8 subjects had ALT levels tested within 

the first 2 years of age, mean ALT 141 U/L (12-1000 U/L). ALT 

levels were elevated in 7/8 (87%) and remained persistently 

elevated in only 2 (51 & 137 U/L) over a mean follow-up of 

2.5 years. Alkaline phosphatase was elevated for age in 7 subjects. 

Other causes of chronic liver disease were excluded in all 

11 subjects. Liver tissue was available on 3/11 and revealed 

cirrhosis in 2 and bridging fibrosis in one subject. Mild to 

moderate steatosis was seen in all biopsies. Scattered PAS 

positive macrophages were seen in biopsies from 2 subjects 

but no inclusion bodies were noted. Transient elastography 

stiffness scores were elevated (17.1, 9.5 and 8.1 Kpa) in the 

3 subjects with advanced fibrosis but normal in the remainder. 

One subject with cirrhosis developed possible HCC at age 1.2 

years and underwent liver transplantation. There was no noted 

association of liver disease progression with specific genotype. 

Conclusion: Chronic liver disease was common among subjects 

with NGLY1 deficiency, particularly during infancy, but 

liver-associated enzymes improved over time reminiscent of 

alpha-1 antitrypsin deficiency. Progression of liver disease to 

cirrhosis was observed and HCC may occur. The pathogenesis 

of the liver disease is currently being evaluated. 

Disclosures: 

The following authors have nothing to disclose: Shilpa Lingala, David E. Kleiner, 

Christina Lam, Lynne A. Wolfe, Carlos R. Ferreira, Donna Krasnewich, William 

A. Gahl, Marc G. Ghany 

1723 

Primary immunodeficiencies in cryptogenic acute liver 

failure in children: a single centre experience 

Rajeev Khanna 1 , Susan Height 3 , Kimberly Gilmour 2 , Nedim 

Hadzic 1 ; 1 Paediatric Liver unit, King’s College Hospital, London, 

United Kingdom; 2 Immunology, Great Ormond Street Hospital, 

London, United Kingdom; 3 Paediatric Haemato-Oncology Unit, 

King’s College Hospital, London, United Kingdom 

Background: Majority of paediatric acute liver failure (PALF) 

cases remain cryptogenic. Haemophagocytic lymphohistiocytosis 

(HLH) is an under-diagnosed clinical syndrome which is 

often associated with primary immunodeficiency disorders. 

HLH is occasionally observed in ALF. Aim: We aimed to identify 

primary immunodeficiency states associated with HLH, 

observed in PALF between January’99 and May’15. Methods: 

We retrospectively analysed our database for children with 

HLH and investigated their demographic, clinical, biochemical 

and immunological markers, management and outcome. 

We used standard international criteria for definition of PALF. 

Children with histological evidence of haemophagocytosis, or 

with deficient proteins involved in lymphocyte granule-mediated 

cytotoxic pathway including perforin, signaling lymphocyte-activating 

molecule-associated protein (SAP), granule 

release assay (CD107a), Munc-13-4, syntaxin 11, syntaxin 

binding protein-2 on flow cytometry were enrolled. Mutational 

analysis was performed for PRF1, UNC13D, STX11, STXBP2, 

RAB27A, XLP-1/SH2D1A, XIAP/BIRC4, CD27 and MAGT1 

genes. Their outcome was compared with historical controls 

with PALF from our centre. Results: We identified 16 children 

with HLH – 8 had histological evidence, whereas 10 had deficient 

proteins (6 perforin, 3 SAP and 1 Munc-13-4). Males 

(81%) predominated. Four (25%) belonged to consanguineous 

families. Ten (62%) were younger than 6 months. Jaundice, 

fever and encephalopathy were present in 12, 12 and 8 cases, 

while hepatomegaly, splenomegaly and lymphadenopathy 

were there in 15, 13 and 4 children, respectively. Median 

peak bilirubin, AST and INR were 158 micromol/L, 1870 IU/L 

and 3.47. Hyperferritinemia, hypertriglyceridemia and hypofibrinogenemia 

were present in 14, 8 and 5 children. Mutations 

were identified in PRF1, SAP/XLP-1 and RAB27A genes in 

7, 3 and 1 patients, respectively. All patients were managed 

as per standard intensive care therapy. Their clinical course 

was complicated by marrow failure, ventilatory requirement, 

systemic inflammatory response syndrome and multi-organ dysfunction 

syndrome in 13, 9, 6 and 5 patients, respectively. 2 

children underwent liver transplantation – one of them died few 

months later due to neurological events. One underwent bone 

marrow transplantation. 1-year mortality was 31% which was 

higher than historical cohort. Conclusion: Identification of this 

group of children with ALF is important for their prognostication 

and family screening. In PALF, diagnosis of HLH needs to 

be pursued by combination of biochemical, histological and 

immunological markers, particularly in young male children 

from consanguineous families. 

Disclosures: 

Nedim Hadzic - Consulting: Alnylam; Speaking and Teaching: Synageva 

The following authors have nothing to disclose: Rajeev Khanna, Susan Height, 

Kimberly Gilmour


1724 

Zinc monotherapy for young pediatric patients with presymptomatic 

Wilson disease: a two-center experience in 

Japan 

Keisuke Eda 1 , Itaru Iwama 2 , Takahito Takeuchi 1 , Yugo Takaki 1 , 

Tatsuki Mizuochi 1 ; 1 Pediatrics, Kurume University School of Medicine, 

Kurume, Japan; 2 Pediatrics, Okinawa Chubu Hospital, 

Uruma, Japan 

Purpose: We previously reported that a reasonable goal in 

treating young pediatric patients with Wilson disease (WD) 

using zinc to be maintaining 24-hour urinary copper excretion 

between 1 and 3 μg/kg/day for the first 1-2 years (Mizuochi, 

et al. JPGN 2011;53:365). Here, we aimed at evaluating 

long-term efficacy and safety of zinc monotherapy for 

young pediatric patients with WD and establishing appropriate 

benchmarks of maintenance therapy. Methods: We performed 

a retro- and prospective study to examine 7 girls (median age 

5 years, range 4-8) who satisfied diagnostic criteria for WD 

and were treated solely with zinc acetate prior to symptom 

onset at Kurume University Hospital and Okinawa Chubu Hospital 

in Japan. No additional WD sequelas, such as jaundice, 

hepatomegaly, or neurologic abnormalities were noted. We 

monitored serum AST and ALT, nonceruloplasmin serum copper, 

and 24-hour urinary copper for 2-7 years after initiation 

of zinc monotherapy. Additional monitorings included white 

blood cell count, hemoglobin, platelet count, serum total bilirubin, 

albumin, iron, amylase, lipase, and prothrombin time, as 

well as 24-hour urinary zinc excretion. We performed abdominal 

ultrasonograpy and evaluated clinical WD manifestations, 

drug compliance, and adverse effects of zinc. The prescribed 

dosage of zinc acetate for patients ≤ 5 years 25 mg twice 

daily; for those children 6 years or older, the dose was 25 mg 

3 times daily. We increased the dosage of zinc if the patients 

had AST/ALT > 50 U/L, and decreased it if they had adverse 

effects of zinc such as iron-deficiency anemia or pancytopenia. 

Results: At the time of diagnosis, AST/ALT and 24-hour urinary 

copper were 207±182/292±211 U/L and 143±68 μg/day 

(7.2±3.5 μg/kg/day), respectively. All patients continued to 

take zinc without any evidence of zinc toxicity. None of our 

7 patients became clinically symptomatic. AST/ALT sharply 

decreased to 35±7/37±16 U/L at 1 year after treatment and 

was mostly maintained less than 50U/L for the remainder of 

the study (AST/ALT: 35±9/42±29 and 33±9/41±17 U/L at 

2 and 6 years after treatment, respectively). Twenty four-hour 

urinary copper decreased to 54±18 μg/day (2.3±0.6 μg/ 

kg/day) at 1 year after treatment and was mostly maintained 

between 1 and 3 μg/kg/day for the remainder of the study 

(1.7±0.7 and 1.9±0.9 μg/kg/day at 2 and 6 years after treatment, 

respectively). Conclusions: Long-term zinc monotherapy 

for young pediatric patients with presymptomatic WD proved 

highly effective and safe. A reasonable goal in treating young 

children with WD using zinc appears to be maintaining both 

AST/ALT under 50 U/L and 24-hour urinary copper excretion 

between 1 and 3 μg/kg/day. 

Disclosures: 

The following authors have nothing to disclose: Keisuke Eda, Itaru Iwama, Takahito 

Takeuchi, Yugo Takaki, Tatsuki Mizuochi 

1725 

Congenital Lactic acidemia (CLA), Amino acidemia (AA), 

Urea cycle defects (UCD) and Organic acidemia (OA) 

presenting with hepatic dysfunction in sick infants and 

children: Diagnosis, Management and Outcome in a 

Pediatric hepatology unit at a specialized liver centre in 

India. 

Seema Alam, Vikrant Sood, Bikrant B. Lal, Dinesh Rawat; Pediatric 

Hepatology, ILBS, New Delhi, India 

CLA, AA, UCD, OA are a group of MLDs which present with life 

threatening illness and various grades of hepatic dysfunction. 

Early recognition and management is of utmost importance in 

countries with limited Newborn Screening Programme. Here 

we present diagnosis, management and outcome children with 

various causes of CLA, UCD and OA following an algorithmic 

approach (Fig 1). Lactate and pyruvate ratio cam differentiate 

between various congenital lactic academia. There were 7 

cases of CLA including 3 cases of Fructose 1, 6 biphosphatemia, 

1 case of respiratory chain defect (RCD) and one case of 

(PEPCK, phosphoeno pyruvate carboxykinase). The three cases 

of Fructose 1, 6 biphosphatemia were managed with fructose 

free diet and recovered completely. The infants with RCD (management 

with Carnitine, Coenzyme Q10, Riboflavine and low 

carbohydrate diet) and PEPCK (supportive management) died. 

A newborn presenting as neonatal liver failure and HCC was 

diagnosed to be Tyrosinemia type 1 and the parents refused 

the option of liver transplant and medical management. A 3 

years old girl with UCD (Partial Ornithine transcarbamyolase 

deficiency) presented as acute liver failure and completely 

recovered with ammonia scavengers and awaiting liver transplant. 

Another UCD presented with hepatic dysfunction at 2 

months of age and died of encephalopathy. The only newborn 

with hepatic dysfunction and Propionic academia improved 

from the metabolic acidosis and ketosis with appropriate diet 

and was sent home with the option of liver transplant in the 

plan. A detailed protocol and algorithmic approach can help 

in early diagnosis and management of these rare liver diseases 

which in turn improves their outcome. 

Figure 1. Protocol based approach to life threatnimg illness with 

hepatic dysfunction. 

Disclosures: 

The following authors have nothing to disclose: Seema Alam, Vikrant Sood, 

Bikrant B. Lal, Dinesh Rawat


1726 

Bile acid metabolism is altered in adolescents with type 

1 diabetes 

Zoehrer Evelyn 1 , Melanie Heckmann 1 , Elke Fröhlich-Reiterer 1 , Hildegard 

Jasser-Nitsche 1 , Günter Fauler 2 , Hubert Scharnagl 2 , Tatjana 

Stojakovic 2 , Jörg Jahnel 1 ; 1 Department of Paediatrics and 

Adolescent Medicine, Medical University Graz, Graz, Austria; 

2 Clinical Institute of Medical and Chemical Laboratory Diagnostics, 

Medical University Graz, Graz, Austria 

Objective: High levels of bile acids (BA) stimulate insulin release 

and therefore decrease serum glucose levels via the TGR5/ 

GLP1 pathway; vice versa, high glucose levels can upregulate 

BA synthesis from cholesterol by stimulating transcription of the 

rate-limiting enzyme Cyp7a1. Type 1 diabetes (T1D) leads to 

hyperglycemia due to lack of endogenous insulin. We hypothesized 

that in children and adolescents with T1D BA levels 

vary coordinately with HbA1c, a longtime marker of glycemic 

control. Design: Concentrations of glucose, HbA1c, and serum 

total BA (tBA) were measured in 86 fasted children and adolescents 

with T1D (age 11 - 20 years) under insulin therapy. 

Patients were divided into three groups: Group 1: low HbA1c 

(< 59 mmol/mol; n=21); Group 2: medium HbA1c (59 – 75 

mmol/mol; n=49) and Group 3: high HbA1c (> 75 mmol/ 

mol; n=16). TBA values were compared with normal values 

for adolescents (n=98; Jahnel et al. 2015, CCLM). Using 10 

μl of serum we determined by high-performance liquid chromatography 

– high-resolution mass spectrometry a BA profile 

including 15 unconjugated and taurine (T)- or glycine (G)-conjugated 

BA; summed, the values for these analytes yield the 

tBA value. Results: In Group 1, with low HbA1c values, mean 

tBA values lay below reference values with 3.1 mmol/l ± 2.4 

mmol/l (3.7 mmol/l ± 1.9 mmol/l). In Group 2, with normal 

HbA1c values, mean tBA values were significantly lower than 

in Group 1 and lay below normal ranges with 2.3 mmol/l ± 

1.8 mmol/l (p


1728 

Serum autoantibodies are associated with chronic hepatitis 

and graft fibrosis after pediatric liver transplantation 

Jeremy K. Rajanayagam 1 , Wolfram Haller 1 , Dominique Debray 2 , 

Henkjan J. Verkade 3 , Valerie A. McLin 4 , Helen M. Evans 5 , Etienne 

M. Sokal 6 , Ekkehard Sturm 7 , Rene Scheenstra 3 , Annette S. Gouw 8 , 

Vladimir Cousin 4 , Sharat Varma 9 , Steffen Hartleif 7 , James Hodson 

10 , Deirdre A. Kelly 1 ; 1 The Liver Unit, BIrmingham Children’s 

Hospital, Birmingham, United Kingdom; 2 Clinique chirurgicale 

Infantile, Necker Hospital, Paris, France; 3 Paediatric Hepatology, 

Beatrix Children’s Hospital, University Medical Center Groningen, 

Groningen, Netherlands; 4 Paediatrics, University Hospitals 

Geneva, Geneva, Switzerland; 5 Hepatology, Starship Children’s 

Hospital, Auckland, New Zealand; 6 Paediatric Hepatology and 

Cell Therapy Lab, Université Catholique de Louvain, Louvain, 

Belgium; 7 Paediatric Gastroenterology & Hepatology, University 

Children’s Hospital, Tuebingen, Germany; 8 Pathology and Medical 

Biology, University Medical Center Groningen, Groningen, 

Netherlands; 9 Cliniques universitaires St Luc, Brussels, Belgium; 

10 University Hospitals Birmingham, Birmingham, United Kingdom 

Background: Chronic hepatitis (CH) and fibrosis are frequent 

features of protocol liver biopsies after pediatric liver transplantation 

(LT). Previously reported associations with positive autoantibodies 

(AuAb) and abnormal immunoglobulins (Ig) suggest 

immunologic mechanisms of graft injury. Aim: To evaluate the 

role of AuAbs and Igs in the etiology of CH and fibrosis. Methods: 

International, multicenter, retrospective analysis of pediatric 

LT recipients with 5 and/or 10 yr protocol liver biopsies 

(normal aminotransaminases), paired serum AuAbs (ANA or 

SMA titre>1:40) and Ig levels, excluding those with pre-LT autoimmune 

liver disease, or other causes of graft injury. Results: 

467 children, from 7 centres underwent LT between 1985- 

2010, predominantly for biliary atresia (n=274), metabolic 

disease (n=58), other cholestatic disorders (n=53), acute liver 

failure (n=44), cryptogenic cirrhosis (n=20) and malignancy 

(n=7). CH was seen in 119/279 (43%) biopsies at 5yrs, 

and 121/229 (53%) biopsies at 10yrs. Fibrosis was present 

in 163/301 (54%) at 5yrs (mild=36%; moderate=12%; 

severe=6%) and 178/226 (79%) at 10yrs (mild=46%;, moderate=20%; 

and severe=13%). Children positive vs negative 

for AuAbs were more likely to exhibit not only CH (5yrs: 59% 

vs 34%, p


Disclosures: 

Kathleen B. Schwarz - Consulting: Novartis; Grant/Research Support: Bristol-Myers 

Squibb, Gilead, Roche/Genentech, Synageva, Vertex, Roche 

The following authors have nothing to disclose: Douglas Mogul, Eric Chow, Xun 

Luo, Allan Massie, Andrew M. Cameron, John F. Bridges, Dorry L. Segev 

Disclosures: 

Philip Rosenthal - Advisory Committees or Review Panels: Abbvie, Retrophin; 

Consulting: Gilead; Grant/Research Support: Gilead, Abbvie, Vertex, BMS, 

Roche/Genetech 

The following authors have nothing to disclose: Emily R. Perito, Robert H. Lustig 

1730 

Pediatric liver transplant recipients (PLTx) have a different 

form of pre-diabetes, with decreased β-cell function 

rather than insulin resistance 

Emily R. Perito 1,2 , Robert H. Lustig 1 , Philip Rosenthal 1,3 ; 1 Pediatrics, 

University of California San Francisco, San Francisco, CA; 2 Epidemiology 

and Biostatistics, University of California San Francisco, 

San Francisco, CA; 3 Surgery, University of California San Francisco, 


Introduction: PLTx have not been previously been evaluated 

for pre-diabetes (defined by the ADA as impaired glucose tolerance 

(IGT; ≥140mg/dL 2 hrs after glucose load) or high 

fasting glucose (>100mg/dL)), despite a known risk with calcineurin-inhibitors 

(CNI). Methods: Cross-sectional study of PLTx 

aged 8-30 yrs, on stable immunosuppression without diabetes, 

with oral glucose tolerance testing (OGTT) by NHANES protocol. 

Patients matched by age, sex, and race/ethnicity with 

≤4 controls from NHANES 2009-2012 cohorts. Results: PLTx 

(n=78) were studied at mean age 15.9 yrs (range 8.1-30.2) 

and mean 11.2 yrs since transplant (range 1-24). PLTx were 

more likely to have IGT (29%,n=72) than matched NHANES 

controls (6.5%,n=292, p


Disclosures: 

The following authors have nothing to disclose: Adam Arterbery, Awo Osafo- 

Addo, Maria Ciarleglio, Yanghong Deng, Mercedes Martinez, Steven J. Lobritto, 

Yaron Avitzur, David Hafler, Markus Kleinewietfeld, Udeme D. Ekong 

1732 

Distance Affects Mortality on the Waitlist in Pediatric 


Joel T. Adler, Yanik Bababekov, James F. Markmann, David C. 

Chang, Heidi Yeh; Massachusetts General Hospital, Boston, MA 

INTRODUCTION: The distance between patients and their liver 

transplant centers contributes to disparities in adult liver transplantation. 

We hypothesized that distance would adversely 

affect the time to transplantation and waitlist mortality for pediatric 

transplant candidates. METHODS: The Scientific Registry 

of Transplant Recipients was queried for isolated pediatric liver 

transplant registrants (under age 18) with chronic liver disease 

and valid ZIP code information from 2003 to 2012. Distance 

from home ZIP code to listing transplant center was calculated 

in miles. Competing events analysis, adjusted for demographic 

factors, rural/urban status, indication, and Pediatric End Stage 

Liver Disease score, was undertaken for transplantation and 

death while on the waitlist. RESULTS: 6,924 children were 

included. The median distance to listing transplant center was 

65 (IQR 17.5-189) miles. Median distance traveled increased 

by listing volume (73.9 vs. 33.8 miles, highest vs. lowest volume 

quartile, P


1734 

Dynamics of pediatric liver allograft histological 

changes and the role of HLA, non-HLA antibodies 

Sharat Varma 3 , Xavier Stephenne 3 , Françoise Smets 3 , Mina 

Komuta 1 , Jerome Ambroise 2 , Catherine de Magnée 4 , Dominique 

Latinne 5 , Raymond Reding 4 , Etienne M. Sokal 3 ; 1 Department of 

anatmopathology, Cliniques universitaires St Luc, Brussels, Belgium; 

2 Centre for Applied Molecular Technologies (CTMA), Institut 

de Recherche Expérimentale et Clinique (IREC),, Université 

Catholique de Louvain (UCL), Brussels, Belgium; 3 Service of Pediatric 

Gastroenterology and Hepatology, Cliniques universitaires 

St Luc, Brussels, Belgium; 4 Department of Pediatric Surgery and 

Transplantation, Cliniques universitaires St Luc, Brussels, Belgium; 

5 Clinical Transplant Immunology Department, Cliniques universitaires 

St Luc, Brussels, Belgium 

Study purpose:HLA and non-HLA antibodies are frequently 

seen post liver transplantation(LT) but their impact on the graft is 

unknown. We studied the impact of these in transplant recipient 

children with minimum confounding factors for causing fibrosis 

and histological changes. Methods: Retrospective study of children 

transplanted between 2004 - 2014 having known HLA 

and non-HLA antibody status and >1 protocol biopsy. Exclusion 

criteria were combined organ transplant, re-transplant, 

chronic rejection, vascular and biliary complications. Biopsies 

were evaluated for bile duct proliferation, portal tract inflammation 

and fibrosis by Metavir and Venturi score.Fibrosis rate 

was calculated as change in fibrosis score divided by duration 

(years) since previous biopsy. HLA antibodies were tested with 

solid single bead Luminex assay with MFI>1500 as cut off. 

Results: 103 children with 323 biopsies were included. Longer 

post LT duration was associated with higher grade of fibrosis 

(p 2000 (clinically relevant). 

We used the sum of class I and class II antibodies to calculate 

total MFI. Results: DSA status in patients with and without ACR 

are summarized in Table 1. Sixty-four patients had DSA measured 

sometime during follow up. Of these, 37 were positive 

for class I or class II at least once. Several patients presented 

with both pre-formed and de novo DSA. DSA were not associated 

with ACR during follow-up. In those patients with measurable 

DSA, MFI >2000 did not correlate with the occurrence 

of ACR. Discussion: In this small, single center retrospective 

study, 57% of children had measurable pre-formed or de novo 

DSA either pre- or post-LT. In spite of this finding, the presence 

of DSA at any time during the early follow up period was not 

associated with an ACR event. . MFI titer did not predict severity 

of rejection among patients who developed ACR.Among 

patients who experienced rejection, MFI titers did not correlate 

with severity of rejection.Prospective studies are required to 

understand better the relationship between DSA and ACR in 

pediatric LT. 

Table 1 

Disclosures: 

The following authors have nothing to disclose: Dominique Schluckebier, Laetitia 

Marie Petit, Vladimir Cousin, Anne-Laure Rougemont, Jean Villard, Dominique 

Belli, Barbara E. Wildhaber, Sylvie Ferrari-Lacraz, Valerie A. McLin


1736 

Surveillance biopsies in pediatric liver transplant recipients 

to guide immunosuppression (IS) reduction 

Amanda J. Posner 1 , Kuang-Yu Jen 3 , Linda Ferrell 3 , Anthony J. 

Demetris 5 , Sandy Feng 4 , Philip Rosenthal 1,4 , Emily R. Perito 1,2 ; 

1 Pediatrics, University of California San Francisco, San Francisco, 

CA; 2 Epidemiology and Biostatistics, University of California San 

Francisco, San Francisco, CA; 3 Pathology, University of California 

San Francisco, San Francisco, CA; 4 Surgery, University of California 

San Francisco, San Francisco, CA; 5 Pathology, University of 

Pittsburgh, Pittsburgh, PA 

Introduction: Recent AASLD guidelines on long-term management 

of pediatric liver transplant (LT) recipients recommend considering 

IS minimization in children with minimal inflammation 

and fibrosis on surveillance biopsy. We instituted surveillance 

biopsy and IS minimization protocols, following experience 

with Immune Tolerance Network (ITN) IS withdrawal trials. 

Methods: Single-center study of surveillance liver biopsies to 

assess eligibility for IS reduction (withdrawal in clinical trial, 

n=27; or minimization to 50% of baseline calcineurin-inhibitor 

[CNI] dose as standard of care [SOC], n=32) in subjects who 

underwent LT prior to age 18. At biopsy, all were ≥5 years 

post-LT on CNI (n=56) or CNI+mycophenolate (n=3). Biopsies 

reviewed by 1 of 2 pathologists using standardized criteria 

from Banff recommendations. Results: 59 pediatric LT recipients 

were included, at median (range) age 13.1 (5.1-22.1) 

years and 9.7 (3.6-20.5) years from LT. LT was whole liver in 

42%, split deceased-donor in 24%, and living-donor in 32%. 

Of 20 subjects with no fibrosis, 75% qualified for IS reduction. 

(FIG) Fibrosis was seen in 39 subjects, but 84% of portal fibrosis 

was Ishak stage 1-2 and 78% of perivenular fibrosis was 

mild. 38/59 (64%) subjects were eligible for IS withdrawal or 

SOC minimization. (FIG) Children eligible vs ineligible were 

younger at transplant (median [IQR] 0.8 [0.5-1.6] vs 1.6 [0.8- 

4.8] years, p=0.03) with no difference in years since transplant, 

whole vs split or living vs deceased donor, AR history, 

or AST/ALT/GGT at biopsy. Of 22 children eligible for SOC 

IS minimization, 5 completed and 5 are in process; none have 

had AR or complications. Conclusion: Surveillance biopsy is 

essential tp inform decision-making on IS reduction. Longitudinal 

studies are needed to establish benefits vs risks of biopsy-guided 

IS reduction. 

Surveillance biopsies and IS reduction eligibility in pediatric LT 

recipients 

Philip Rosenthal - Advisory Committees or Review Panels: Retrophin, Gilead; Consulting: 

Roche, Abbvie; Grant/Research Support: Roche, Bristol MyersSquibb, 

Gilead, Vertex, Abbvie 

The following authors have nothing to disclose: Amanda J. Posner, Kuang-Yu Jen, 

Linda Ferrell, Anthony J. Demetris, Emily R. Perito 

1737 

Epidemiology and outcomes of pediatric liver transplant 

recipients with multidrug resistant bacterial and fungal 

infections 

Gillian Noel, Mark J. Abzug, Donna Curtis, Zhaoxing Pan, Shikha 

Sundaram; University of Colorado/Children’s Hospital Colorado, 

Denver, CO 

Infection is a significant source of morbidity and mortality 

following liver transplantation. Adult transplant recipients 

experience increased colonization and infection with drug-resistant 

organisms including methicillin-resistant staphylococcus 

(MRSA), extended-spectrum beta-lactamase producers, vancomycin-resistant 

enterococci, and multidrug resistant pseudomonas 

aeruginosa. Limited data exist regarding the incidence of 

multidrug resistant organisms in the pediatric liver transplant 

(LT) population. Objective: To define the incidence and impact 

on outcomes of multidrug resistant organisms in pediatric LT 

recipients. Methods: A retrospective cohort study of pediatric LT 

recipients at Children’s Hospital Colorado between Jan 2006 

and Dec 2014 was conducted. Demographic, clinical and 

outcome data were collected. Results: Sixty subjects were identified 

and followed for one year post-LT (mean age at LT of 4.9 

± 6.1 years; 55% female, 67% white, 51% transplanted due 

to biliary atresia). Treatment for suspected bacterial infections 

occurred in 51% of subjects, but only 44% had a documented 

isolate. Major isolates included coagulase negative staphylococci, 

enterococci, and gram-negative enterics. Although 

MRSA colonization was common (6.5% MRSA positive at LT), 

MRSA infections did not occur. Two percent of subjects had 

multidrug resistant organisms (extended spectrum beta lactamase 

producers) identified. Of subjects treated for bacterial 

infections, 68% received 1 course, 18% received 2-3 courses, 

and 14% received 4 or more courses of antibiotics. Presumed 

cholangitis and bacteremia were the most common reasons for 

antibiotic treatment. Treatment for fungal infections occurred in 

5% of subjects, although no multidrug resistant fungal infections 

were identified. All treated subjects received one course of 

antifungal treatment. Of those treated, only 50% had a documented 

isolate, primarily candidal species. Fungemia was 

the most common reason for antifungal treatment. Subjects 

with clinically suspected bacterial or fungal infections post-LT 

had more ICU admissions (1-3) versus those without suspected 

infection (0) post-LT (p=0.05). There were no differences in 

overall numbers of hospitalizations, reoperation rates, rejection, 

graft loss, or death in the twelve months post-LT as related 

to infection status. Conclusions: While serious bacterial and 

fungal infections commonly occur in pediatric liver transplant 

recipients, multidrug resistant infections are uncommon in our 

institution. Infected subjects have more complicated courses, 

characterized by ICU admissions, but their overall outcomes 

are similar to those without infections. 

Disclosures: 

The following authors have nothing to disclose: Gillian Noel, Mark J. Abzug, 

Donna Curtis, Zhaoxing Pan, Shikha Sundaram 

Disclosures: 

Sandy Feng - Consulting: Novartis


1738 

Immunization Practices Amongst Pediatric Transplant 

Hepatologists at SPLIT Centers 

Amy G. Feldman 1,4 , Shikha Sundaram 1,4 , Brenda Beaty 3 , Allison 

Kempe 2 ; 1 Pediatric Gastroenterology, Hepatology and Nutrition, 

Children’s Hospital Colorado, Aurora, CO; 2 General Pediatrics, 

Children’s Hospital Colorado, Aurora, CO; 3 Adult and Child Center 

for Health Outcomes Research and Delivery Science, Aurora, 

CO; 4 University of Colorado School of Medicine, Aurora, CO 

Objectives: To assess amongst US and Canadian pediatric 

transplant hepatologists: (1) current immunization practices 

before and after liver transplantation and (2) beliefs about the 

safety of different vaccines before and after liver transplantation. 

Study Design: An 80 item email survey of US and Canadian 

pediatric transplant hepatologists at centers participating 

in the Studies of Pediatric Liver Transplantation (SPLIT) consortium 

from December 2014 to March 2015. Results: The overall 

response rate was 80%, representing 97% of SPLIT centers. 

Thirty-four percent of programs did not have written protocols 

to guide immunizations before and after transplant and 50% 

of programs always involved an infectious disease physician 

as part of the formal liver transplant evaluation. Administration 

of Palivizumab was considered by 45% of hepatologists 

pre-transplant and by 64% post-transplant. Only 25% of hepatologists 

ever recommended live vaccines after transplant. Timing 

of inactive vaccines after transplant ranged from 1 month 

to greater than 6 months post-transplant. Only 6% of programs 

were able to provide all vaccines to patients in hepatology/ 

transplant clinic pre and post-transplant. Similarly, 6% of programs 

were always able to provide influenza vaccine to family 

members/household contacts. Conclusions: There is wide variation 

in immunization practices before and after liver transplant 

amongst pediatric transplant hepatologists. Specific evidence 

based protocols are needed to guide immunization practices 

before and after transplant and ensure appropriate vaccination 

of pediatric liver transplant recipients. 

Disclosures: 

The following authors have nothing to disclose: Amy G. Feldman, Shikha Sundaram, 

Brenda Beaty, Allison Kempe 

1739 

Detectable HBVDNA in liver allograft tissues has no clinical 

significance on long term follow up in liver transplant 

recipients under HBIg plus Antiviral prophylaxis 

Deniz Mut 1 , Ilker Turan 1 , Deniz Nart 2 , Funda Yilmaz 2 , Selda Erensoy 

1 , Ulus S. Akarca 1 , Galip Ersoz 1 , Fulya Gunsar 1 , Zeki Karasu 1 ; 

1 Department of Gastroenterology, Ege University, Izmir, Turkey; 

2 Pathology, Ege University, Izmir, Turkey 

Combination prophylaxis with hepatitis B immune globulin 

(HBIG) plus an oral anti-viral (OAV) is widely accepted regimen 

for the prevention of recurrent hepatitis B virus (HBV) infection 

after liver transplantation, although there is no consensus on 

dose of HBIg. Persistency of HBV infection in allograft tissue in 

spite of prophylaxis is speculative. We examined the presence 

of HBVDNA using real-time polymerase chain reaction (PCR) in 

liver allografts in liver transplant recipients who undergone liver 

transplantation because of HBV related liver disease. Patients 

who had HBIg+Lam prophylaxis and who have no detectable 

serum virologic markers (HBsAg and/or HBV DNA) for recurrent 

disease during follow up of at least 3-years were included 

study. Histologic examination is also performed. We investigated 

if there is any correlation between tissue HBVDNA and 

presence of pretransplant HCC, Delta co-infection, the type 

of graft donor (living/cadaveric), the type of post transplant 

immunosuppressive medication. Results: One-hundred and fifty 

(116 male, 34 female) patients with a mean age of 47-years 

(18-64 years) were included into the study. Median duration 

of follow up was 5 (3-9) years after transplantation. Sixty-four 

patient had delta co-infection and 44 patients had HCC before 

transplantation. Of the patients, 108 (72%) had their grafts 

from living donors. HBVDNA was detectable by PCR in liver 

tissue of 18 (12%) patients. Immunostaining for HBVwas negative 

on liver histology of all patients. Of the 18 patients with 

detectable HBVDNA by PCR, the liver graft showed no hepatitis 

on histologic examination. Detectable tissue HBVDNA had 

no correlation with age, sex, delta co-infection, HCC, type of 

donor and type of immunosuppressive medication. Conclusion: 

in spite of combination prophylaxis with HBIg+OAV, and of 

there is no detectable serologic markers in their serum, some 

(12%) cases may have detectable HBVDNA in their liver tissue. 

However presence of HBVDNA in grafts has no clinical impact. 

But in patients with detectable HBVDNA, the risk of reactivation 

may persist indefinitely. 

Disclosures: 


AbbVie 

The following authors have nothing to disclose: Deniz Mut, Ilker Turan, Deniz 

Nart, Funda Yilmaz, Selda Erensoy, Galip Ersoz, Fulya Gunsar, Zeki Karasu 

1740 

HBV cccDNA evaluation in HBV-core antibody 

(HBcAb)-positive liver transplant donors 

Gian Paolo Caviglia 1 , Francesco Tandoi 2,3 , Maria Lorena Abate 1 , 

Alessia Ciancio 1,4 , Renato Romagnoli 2,3 , Antonina Smedile 1,4 ; 

1 Medical Sciences, University of Turin, Turin, Italy; 2 Surgical Sciences, 

University of Turin, Turin, Italy; 3 Liver Transplant Center, 

Città della Salute e della Scienza, Molinette Hospital, Turin, Italy; 

4 Gastroenterology, Città della Salute e della Scienza, Molinette 

Hospital, Turin, Italy 

Introduction Occult hepatitis B virus infection (OBI) is defined 

as the presence of Hepatitis B Virus (HBV) DNA in the liver of 

subjects negative for hepatitis B surface antigen (HBsAg). A 

high OBI prevalence has been reported in the anti-HBV core 

antibody (HBcAb)-positive population as residue of a past 

infection. OBI-positivity may be responsible for recurrent or de 

novo hepatitis B in immunosuppressed subjects. In the setting 

of liver transplant, prophylaxis for hepatitis B recurrence is 

routinely performed in all HBcAb-positive liver graft recipients. 

However, the risk of HBV reactivation is linked to the presence 

of intrahepatic covalently-closed-circular DNA (cccDNA) being 

the replication-competent form. Aim To evaluate the presence 

of intrahepatic HBV cccDNA and its correlation with OBI status 

in a cohort of HBcAb-positive liver donors. Methods Liver 

biopsies from 78 consecutive HBsAg-negative/HBcAb-positive 

deceased donors (mean age 66 [31-91] years; M/F=50/28) 

were tested for OBI by 4 parallel nested-PCRs to detect HBV S, 

Core, Polymerase and X sequences. According to Taormina 

expert meeting statements, samples turned positive for at least 

2 targets were scored as OBI-positive. cccDNA presence was 

assessed by a specific nested-PCR after enzymatic digestion 

with S1 nuclease. Results Among HBcAb-positive liver donors, 

OBI prevalence was 56.4% (44/78). In detail, 28 liver biopsies 

were 4/4 targets positive, 7 were 3/4, 9 were 2/4, 11 were 

1/4 while 23 biopsies were completely negative. cccDNA 

prevalence was 32.1% (25/78) in the whole cohort and 

45.5% (25/55) in the OBI-positive group (including those positive 

for only 1 target). A significant direct correlation was found 

between OBI and cccDNA-positivity (r=0.594; p


ed-PCRs was significantly associated with cccDNA detection 

(OR=2.295, 95%CI 1.7574-4.5556; p10 ng/mL, 83% were male, and 

88% were non-Asian. Comorbidities included diabetes (29%), 

smoking (63%), HBV (2%), and HIV (12%). Median HCC size 

was 2.8 cm (range, 0.8-18.2), 83% had a single lesion on 

imaging and 51% had histologic vascular invasion. Median 

lab values at the time of HCC diagnosis indicated that liver 

function was generally well-preserved: albumin 4.2 g/dL (2.1- 

5.0), platelets 148 x 10 3 cells/μL (48-446), and total bilirubin 

1.3 mg/dL (0.2-8.8). HCC was diagnosed via surveillance 

(imaging and/or AFP) in 27 patients, and in response to symptoms 

in 13. In the surveillance group, 85% were within Milan 

criteria vs 23% in the symptomatic group (p


The following authors have nothing to disclose: Chiara Rocha, M. Isabel Fiel, Erin 

H. Doyle, Nicolas Goossens, Yujin Hoshida, Myron E. Schwartz 

1743 

Treating HCV patients on the LT waiting list with Sofosbuvir 

plus Ribavirin is associated with clinical benefits 

and allows for safe assignement of older grafts to LT 

recipients 

Maria Francesca Donato 1 , Daniele E. Dondossola 2 , Federica Malinverno 

1 , Sara Monico 1 , Cristina Rigamonti 1 , Lucio Caccamo 2 , 

Alessio Aghemo 1 , Margherita Cavenago 2 , Giovanna Lunghi 3 , 

Barbara B. Antonelli 2 , Giorgio Rossi 2 , Massimo Colombo 1 ; 1 Division 

of Gastroenterology and Hepatology, Fondazione IRCCS 

Ca’ Granda Ospedale Maggiore Policlinico, University of Milan, 

Milan, Italy; 2 Division of Surgery and Liver Transplant, Fondazione 

IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy; 

3 Department of Clinical Chemistry and Microbiology, Bacteriology 

and Virology Units, Fondazione IRCCS Ca’ Granda Ospedale 

Maggiore Policlinico, Milan, Italy 

Background and aim SOF/R treatment in HCV(+) cirrhotics 

on the liver transplant (LT) waiting list is a major breakthrough 

since HCV-graft recurrence might be prevented in 95% of 

recipients with >4 weeks viral suppression before transplant. 

We aimed to evaluate the clinical-virological outcome of 

pre-LT SOF/R regimen in patients who stopped treatment at 

LT because they were HCV-RNA undetectable for >4 weeks 

(SOF-pT) comparing them to patients who received bridging 

pre to post-transplant therapy (SOF-bppT) for lack of 4 weeks 

of HCV RNA undetectability at LT. Methods From July 2014 

an Italian compassionate use program by Gilead-Sciences 

endorsed by AISF-AIFA allowed us to treat all HCV consecutive 

transplant candidates at our Center with SOF 400 mg/day + 

R 200-1000 mg/day. Treated patients were compared with 

19 previously transplanted HCV-recipients (January-June 2014) 

who were left untreated before LT. Results 30 HCV-LT candidates 

received SOF/R pre-LT: 16 received LT while 13 are still 

in LT list and 1 died. 9/16 (56%) received SOF-pT whereas 7 

(44%) received SOF-bppT for a total duration of 24 weeks. The 

2 groups of SOF-treated patients were similar for sex, recipient 

age, donor age, HCV genotype or pretreatment viremia, 

MELD/Child and type of immunosuppression. The duration of 

SOF-therapy before LT was shorter in SOF-b-ppT compared to 

SOF-pT (32 vs 85 days, respectively; p


Varuna Aluvihare - Speaking and Teaching: Astellas 

John G. O’Grady - Advisory Committees or Review Panels: Astellas, Novartis; 

Speaking and Teaching: Astellas 




The following authors have nothing to disclose: Anna Mrzljak, Sarah Knighton, 

Matthew J. Bruce, Mary Horner, Aisling B. Considine, Michael A. Heneghan, 

Abid Suddle 

1745 

An Evidence-Based Algorithm to Optimize Patient Selection 

among Class III Obese Liver Transplant Waitlist 

Registrants 

Ryan B. Perumpail 1 , Andy Liu 2 , Channa R. Jayasekera 1 , James 

H. Marcus 4,5 , Swetha Tummala 1 , Sammy Saab 3 , Zobair M. 

Younossi 4,5 , Robert J. Wong 6 , Aijaz Ahmed 1 ; 1 Division of Gastroenterology 


Palo Alto, CA; 2 Medicine, Albert Einstein College of Medicine, 

Bronx, NY; 3 Medicine and Surgery, David Geffen School of 

Medicine at the University of California at Los Angeles, Los Angeles, 

CA; 4 Center for Liver Diseases, Inova Fairfax Hospital, Falls 


Inova Health System, Falls Church, VA; 6 Division of Gastroenterology 

and Hepatology, Alameda Health System - Highland Hospital 

Campus, Oakland, CA 

Background: Class III obesity (MOB), body mass index (BMI) 

≥40 kg/m2, is increasing in prevalence among U.S. waitlist 

registrants for liver transplantation (LT). MOB is considered a 

relative/absolute contraindication for LT at many centers and 

by some health insurers due to increased risk of perioperative 

morbidity. However, MOB is not independently associated 

with lower long-term survival following LT. An evidence-based 

algorithm is needed to optimize patient selection for LT in the 

setting of MOB. Aim: To establish an evidence-based algorithm 

to optimize patient selection for LT in patients with MOB. 

Methods: We conducted a retrospective cohort study using 

U.S. national data from the United Network for Organ Sharing 

registry from 2003-2012 (MELD era) to evaluate the impact of 

African American (AA) race, chronic hepatitis C virus (HCV) 

infection, diabetes mellitus (DM), hepatocellular carcinoma 

(HCC), and the presence of ascites on long-term survival 

among adult LT recipients with MOB. Survival following LT was 

evaluated with Kaplan Meier methods. Multivariate Cox proportional 

hazards models were used to evaluate the independent 

impact of the predetermined variables on post-LT survival. 

The model was adjusted for age, sex, race/ethnicity, year of 

LT, hepatic encephalopathy, Model for End-Stage Liver Disease 

score, cardiac disease, HCV status, DM, HCC, and the presence 

of ascites. Identified independent predictors of lower survival 

were incorporated into a stepwise algorithm for LT patient 

selection. Results: Overall, 1,845 adult patients with MOB 

underwent LT. When compared to non-HCV patients, 5-year 

survival in HCV patients was lower (68.3% vs 75.0%; 95% CI, 

63.4%- 72.7% vs. 70.8%-78.7%; p


Disclosures: 











The following authors have nothing to disclose: James Fung, Tiffany C. Wong, 

Cindy K. Cheung, Kenneth S. Chok, Wing-chiu Dai, Tan To Cheung, William 

Sharr, Albert Chan, Sui-Ling Sin, See-Ching Chan, Chung-Mau Lo 









Alessio Aghemo - Advisory Committees or Review Panels: Gilead Sciences, MSD, 

AbbVie, Janssen; Speaking and Teaching: Jannsen, MSD, Abbvie, BMS, Gilead 

Sciences 

The following authors have nothing to disclose: Stella De Nicola, Maria F. 

Donato, Caterina Premoli, Giovanna Lunghi, Patrizia Bono, Matteo A. Manini, 

Federica Malinverno, Sara Monico, Paolo Reggiani 

1747 

Prevalence and Clinical Impact of Hepatitis E Virus in 

Immunosuppressed Patient after Liver Transplantation 

Stella De Nicola 1 , Maria F. Donato 1 , Caterina Premoli 1 , Giovanna 

Lunghi 2 , Patrizia Bono 2 , Matteo A. Manini 1 , Pietro Lampertico 1 , 

Federica Malinverno 1 , Sara Monico 1 , Paolo Reggiani 3 , Massimo 

Colombo 1 , Alessio Aghemo 1 ; 1 Division of Gastroenterology and 

Hepatology, Fondazione IRCCS Ca’ Granda Ospedale Maggiore 

Policlinico, University of Milan, Milan, Italy; 2 Department of Clinical 

Chemistry and Microbiology, Bacteriology and Virology Units, 

Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, 

Milan, Italy; 3 Division of Surgery and Liver Transplant, Fondazione 

IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy 

Backgruond: HEV infection is a rising health problem, as it is 

the cause of acute and chronic hepatitis especially in immunosuppressed 

patients. The prevalence of HEV infection in 

Italy and its clinical impact on liver transplanted (LT) patients 

is unknown. Aim: To investigate the seroprevalence of IgG-IgM 

anti-HEV in LT patients and to compare it with patients with 

chronic HCV or HBV infection, or healthy Volunteers. Moreover 

we evaluated the impact of HEV in transplanted patients 

on overall survival, rejection rates, fibrosis progression and 

incidence of extra hepatic manifestastions. Methods: Serology 

for HEV IgG and IgM was done with Wantai test (ELISA), HEV 

RNA (in-house RT-PCR) was tested in all LT patients in at least 

two timepoints during the first year following liver transplantation. 

Results: 79 patients who underwent liver transplantation 

at our center between 2010 and 2014 were enrolled. 

The median age at transplantation was 55 years, 11 (14%) 

patients were of foreign origins and HCV was the main etiology 

57%. 26 transplanted patients (33%) tested positive for 

IgG anti HEV, 3 patients, who were IgG anti-HEV negative 

at LT developed IgG during the period of observation. None 

of 79 patients were positive for HEV RNA. The seroprevalence 

in HCV patients 19% (19/100) (p=0.03), HBV patients 

12% (12/100) (p=0.0009) and healthy controls 5% (5/199) 

(p=0.0001) were significantly lower than in LT patients. The 

prevalence was not influenced by fibrosis stage in HBV or 

HCV patients. Overall none of the 79 patients had chronic 

HEV infection. In terms of clinical endpoints in the LT patients, 

no difference was seen in terms of survival after transplantation, 

onset of diabetes, onset of kidney disease, onset of 

neurological syndromes or fibrosis progression in IgG positive 

versus IgG negative patients. Conclusion: Anti-HEV IgG are 

common in the Italian general population and even more so in 

LT patients or patients with chronic HCV or HBV infection. However, 

in our cohort we do not report a major impact in terms of 

clinical endpoints in LT patients probably as a consequence of 

the 0% rate of chronic HEV infection. 

Disclosures: 

Pietro Lampertico - Advisory Committees or Review Panels: Bayer, Bayer; Speaking 

and Teaching: Bristol-Myers Squibb, Roche, GlaxoSmithKline, Novartis, Gilead, 

Bristol-Myers Squibb, Roche, GlaxoSmithKline, Novartis, Gilead 

1748 

Long-term outcomes after hepatic resection for hepatocellular 

carcinoma in patients with sustained virological 

responses to interferon therapy for chronic hepatitis C 

Shogo Tanaka 1 , Shigekazu Takemura 1 , Akihiro Tamori 2 , Masahiko 

Kinoshita 1 , Norifumi Kawada 2 , Shoji Kubo 1 ; 1 Department of 

Hepato-Biliary-Pancreatic Surgery, Osaka City University Graduate 

School of Medicine, Osaka, Japan; 2 Department of Hepatology, 

Osaka City University Graduate School of Medicine, Osaka, 

Japan 

Background: The long-term outcome after hepatic resection for 

hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC) 

in patients who had previously achieved sustained virological 

responses (SVR) to interferon (IFN) therapy and who achieved 

SVR to adjuvant IFN therapy has been inclusive. Method: 

The clinical records of 277 patients who underwent hepatic 

resection for HCV-related early stage HCC between 1993 

and 2012 were retrospectively reviewed. Thirty-seven of the 

277 patients had achieved SVR at the detection of HCC (pre- 

SVR group). Twenty-three patients achieved SVR after hepatic 

resection (post-SVR group). The control group included the 

remaining 217 patients who had received IFN therapy but had 

not achieved SVR or who had not received IFN therapy. We 

investigated the effect of SVR on the postoperative recurrence 

and survival. Results: Twenty-two patients (59%) in the pre-SVR 

group, 23 (65%) in the post-SVR group, and 164 (59%) in the 

control group had postoperative recurrence during the follow-up 

periods, respectively (p=0.09). The disease-free survival rate 

was significantly better in the pre-SVR group (5/10/15 years: 

49%/29%/29%) and the post-SVR group (61%/36%/27%) 

than the control group (23%/7%/7%, p=0.000054). The 

overall survival rate at 10/15 years after hepatic resection 

was 68%/68% in the pre-SVR group, 78%/78% in the post- 

SVR group, and 13%/11% in the control group, respectively 

(p


pre- and post-operative achievement of SVR had better survival 

rates after hepatic resection for HCV-related HCC. 

Disclosures: 



The following authors have nothing to disclose: Shogo Tanaka, Shigekazu 

Takemura, Akihiro Tamori, Masahiko Kinoshita, Shoji Kubo 

1749 

Effects of pre-transplant hepatitis C clearance on liver 

transplantation listing and wait time – a single center 

experience 

Po-Hung Chen 1 , Mary G. Bowring 2 , Lauren M. Kucirka 2 , Christine 

M. Durand 1 , Andrew Ofosu 2 , Alia S. Dadabhai 1 , James P. Hamilton 

1 , Andrew M. Cameron 3 , Dorry L. Segev 3 , Mark S. Sulkowski 1 , 

Ahmet Gurakar 1 ; 1 Medicine, Johns Hopkins University, Baltimore, 

MD; 2 Johns Hopkins University, Baltimore, MD; 3 Surgery, Johns 

Hopkins University, Baltimore, MD 

Background: The use of hepatitis C virus (HCV)-positive 

allografts in viremic recipients helps expand the pool of 

potential donors – especially at high usage LT centers – but 

these organs are generally inappropriate for patients cleared 

of chronic HCV. Despite recent advancements using new 

direct-acting antivirals (DAA), no clear guidelines yet exist 

regarding the optimal timing of HCV treatment peri-LT. The 

present analysis aims to assess the associations of successful 

pre-LT HCV treatment in the era of interferon-sparing DAA regimens 

with LT listing and time on wait list. Methods: Data were 

collected retrospectively for consecutive adults with HCV-related 

liver disease who underwent cadaveric LT at the Johns Hopkins 

Hospital from January 1, 2014 to April 5, 2015. Recipients 

were grouped as “HCV Negative” or “HCV Positive” based on 

serum HCV RNA status at LT. The decision to treat HCV pre-LT 

was made by individual providers. Between-group comparisons 

were made of key recipient and donor characteristics. 

Results: There were 50 cadaveric LT for HCV-related liver disease, 

32 (64%) of which were in “HCV Positive” group. Within 

this group, 13 (41%) HCV-positive allografts were used. No 

significant differences were noted between “HCV Negative” 

and “HCV Positive” groups in age, gender, race, presence 

of hepatocellular carcinoma, and MELD at organ allocation. 

Among “Negative” recipients 11 (61%) had blood type O, 

versus 12 (38%) in “Positive” group. Sixteen (89%) “Negative” 

recipients were alive at 1 year post-LT, compared to 29 

(91%) “Positive” patients (P = 0.8). (Table) Finally, concordant 

HCV-negative recipient-donor pairs waited a median of 

158 days (Q25-Q75: 13-296) before LT; HCV-positive pairs 

waited 221 (26-241) days (P = 0.7). Conclusions: There were 

no significant between-group differences, including time on 

wait list, among recipients regardless of treatment with DAA 

pre-LT. The use of HCV-positive organs in viremic recipients did 

not adversely impact survival at one year. Larger studies are 

needed to confirm whether these relationships hold true across 

the entire spectrum of MELD and blood types at LT listing. 

Disclosures: 



BMS 

Ahmet Gurakar - Advisory Committees or Review Panels: Gilead; Grant/ 


The following authors have nothing to disclose: Po-Hung Chen, Mary G. Bowring, 

Lauren M. Kucirka, Christine M. Durand, Andrew Ofosu, Alia S. Dadabhai, 

James P. Hamilton, Andrew M. Cameron, Dorry L. Segev 

1750 

Protective Birth Cohort Effect in Baby Boomers Despite 

Presence of Poor Predictors 

Ryan B. Perumpail 1 , Andy Liu 2 , Channa R. Jayasekera 1 , Zobair M. 

Younossi 3,4 , Robert J. Wong 5 , Aijaz Ahmed 1 ; 1 Division of Gastroenterology 


Palo Alto, CA; 2 Medicine, Albert Einstein College of Medicine, 

Bronx, NY; 3 Department of Medicine, Center for Liver Diseases, 

Inova Fairfax Hospital, Falls Church, VA; 4 Betty and Guy Beatty 

Center for Integrated Research, Inova Health System, Falls Church, 

VA; 5 Division of Gastroenterology and Hepatology, Alameda 

Health System, Highland Hospital, Oakland, CA 

Background: Hepatitis C virus (HCV) is a leading etiology of 

chronic liver disease in the U.S. Seventy-five percent of HCV 

patients are of the baby boomer (BB) generation (born 1945- 

1965). Our study aims to analyze the impact of the BB generation 

on survival following liver transplantation (LT) in the 

U.S. Methods: We conducted a retrospective cohort study 

using population-based national data from the United Network 

for Organ Sharing 1995-2012 registry to evaluate birth 

cohort-specific (BB vs. non-BB) and etiology-specific (HCV vs. 

non-HCV) impact on outcomes. Overall post-LT survival was 

stratified by birth cohort and HCV status, and was evaluated 

with Kaplan Meier methods and multivariate Cox proportional 

hazards models. Results: From 1995 to 2012, the proportion 

of LT waitlist registrations represented by BB patients increased 

from 47.4% to 77.0%. These trends were primarily driven by 

HCV, and BB patients accounted for 90% of HCV-related LT 

waitlist registrants in 2012. Similar trends were seen among LT 

recipients: BB patients accounted for 78.1% of all LT in 2012 

compared to 21.9% among non-BB patients. Furthermore, BB 

patients accounted for 91.1% of HCV-related LT. Among both 

HCV and non-HCV patients, BB patients had significantly better 

5-year post-LT survival (HCV: 69.5% vs. 64.2%, p< 0.001; 

non-HCV: 79.8% vs. 74.2%, p


Disclosures: 






Inc. 

The following authors have nothing to disclose: Ryan B. Perumpail, Andy Liu, 

Channa R. Jayasekera, Robert J. Wong 

1751 

Prevention of post-transplant HCV recurrence by a 24 

week regimen of Sofosbuvir/ribavirin (SOF/R) bridging 

pre-post transplant (b-ppT): a real life strategy 

Maria F. Donato 1 , Sonia Berardi 2 , Rosa Maria Iemmolo 3 , Luca 

S. Belli 4 , Marzia Montalbano 5 , Sherrie Bhoori 6 , Giulia Pieri 7 , 

Ilaria Lenci 8 , Alessandra Riva 9 , Veronica Bernabucci 10 , Sara 

Monico 1 , Chiara Mazzarelli 4 , Federica Malinverno 1 , Daniele E. 

Dondossola 11 , Chiara Taibi 5 , Giorgio Rossi 11 , Antonio Daniele 

Pinna 17 , Fabrizio Di Benedetto 3 , Luciano De Carlis 13 , Giuseppe 

M. Ettorre 14 , Vincenzo Mazzaferro 6 , Umberto Montin 12 , Giuseppe 

Tisone 15 , Marco Vivarelli 16 , Maria Rosa Tamè 2 , Paolo Caraceni 18 , 

Maria Cristina Morelli 2 ; 1 Division of Gastroenterology and Hepatology, 

Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, 

Università degli Studi di Milano, Milan, Italy; 2 Department 

of Digestive Disease and Internal Medicine, St Orsola-Malpighi 

University Hospital, Bologna, Italy; 3 Liver and Multivisceral Transplant 

Center, Azienda Ospedaliero Universitaria di Modena, 

Modena, Italy; 4 Hepatology and Gastroenterology Unit, Ospedale 

Niguarda Cà Granda, Milano, Italy; 5 Hepatology and Infectious 

Diseases Unit, National Institute for Infectious Diseases “L. Spallanzani”, 

Roma, Italy; 6 Gastroenterology, Surgery and Liver Transplantation, 

Fondazione Istituto Nazionale Tumori IRCCS, Milan, 

Italy; 7 Division of Hepatology, IRCCS AO San Martino-IST-Genova, 

Genova, Italy; 8 Hepatology Unit, Università Tor Vergata Roma, 

Roma, Italy; 9 Division of Infectious Diseases, AOU Ospedali Riuniti 

Ancona, Ancona, Italy; 10 Division of Gastroenterology, Azienda 

Ospedaliero Universitaria di Modena, Modena, Italy; 11 Division of 

General Surgery and Liver Transplantation, Fondazione IRCCS Ca’ 

Granda Ospedale Maggiore Policlinico, Milan, Italy; 12 Epatobiliary 

and Liver Transplantation Surgery, AO Universitaria Integrata 

Verona, Verona, Italy; 13 Division of General Surgery and Liver 

Transplantation, Ospedale Niguarda Ca’ Granda, Milan, Italy; 

14 Division of General Surgery and Liver Transplantation, National 

Institute for Infectious Diseases “L. Spallanzani”, Roma, Italy; 

15 Department of Experimental Medicine and Surgery, Università 

Tor vergata Roma, Roma, Italy; 16 Hepatobiliary and Abdominal 

Transplantation Surgery, AOU Ospedali Riuniti Ancona, Ancona, 

Italy; 17 Liver Surgery and Transplantation Unit, St Orsola-Malpighi 

University Hospital, Bologna, Italy; 18 Division of Internal Medicine, 

St Orsola-Malpighi University Hospital, Bologna, Italy 

Background and aims: SOF/R treatment in HCV(+) cirrhotics 

waiting liver transplant (LT) is a major breakthrough since HCVgraft 

recurrence might be prevented in 95% of recipients with 

>4 weeks viral suppression before transplant. We aimed to 

evaluate if bridging SOF/R from pre to post-transplant phase 

(b-ppT) could be feasible, safe and prevent HCV-graft recurrence 

also in those HCV patients without HCV-RNA negativity 

for at least 4 weeks before LT. Methods: SOF 400 mg/day+R 

200-1200 mg/day was given for a total duration of 24 weeks 

pre-post-LT. Results: 25 HCV-LT candidates (2re-LT) received 

b-ppT SOF within an Italian compassionate use program (ITA- 

COPS study). LT indication was HCC in 17 (MELD 8-21) and 

decompensated cirrhotics in 8 (MELD 9-26). HCV-genotype 

was 1 or 4 in 18. At transplant: 14 patients were HCV-RNA(+) 

with a median viral load of 35 IU/ml (12-2584) and 11 were 

HCV-RNA undetected for less than 4 weeks following a median 

treatment of 36 days of SOF/R (5-112). Post-transplant: 16 

patients transiently discontinued SOF after-LT (median=1 day), 

but all cleared HCV within week-4 post-LT. At the time of the 

analysis, all recipients were HCV-free and alive, with a median 

post-transplant follow-up of 26 weeks (7-98). Eighteen recipients 

(72%) completed SOF/R treatment with SVR4 and SVR12 

rates of 100% (17/17 and 9/9, respectively). One patient was 

re-transplanted; 7 developed an episode of AKI; 3=arytmia; 

1=sepsis; 4 (Cya-group) cellular rejection requiring switch to


Tacrolimus/steroid boli. Conclusions: A b-ppT SOF/R-regimen 

in HCV-listed cirrhotics with suboptimal virological response at 

the time of transplant, might represent an optimal strategy to 

avoid post-transplant graft reinfection. 

Disclosures: 

Sherrie Bhoori - Speaking and Teaching: Bayer, BTG 

Vincenzo Mazzaferro - Advisory Committees or Review Panels: Bayer; Grant/ 

Research Support: BTG 



The following authors have nothing to disclose: Maria F. Donato, Sonia Berardi, 

Rosa Maria Iemmolo, Luca S. Belli, Marzia Montalbano, Giulia Pieri, Ilaria Lenci, 

Alessandra Riva, Veronica Bernabucci, Sara Monico, Chiara Mazzarelli, Federica 

Malinverno, Daniele E. Dondossola, Chiara Taibi, Giorgio Rossi, Antonio 

Daniele Pinna, Fabrizio Di Benedetto, Luciano De Carlis, Giuseppe M. Ettorre, 

Umberto Montin, Giuseppe Tisone, Marco Vivarelli, Maria Rosa Tamè, Maria 

Cristina Morelli 

1752 

Hepatitis E infection in immunocompromised persons in 

Argentina 

Jose Debes 1 , Maribel Martinez Wassaf 2 , Maia Belen Pisano 3 , Leonardo 

Marianelli 4 , Martin Lotto 3 , Domingo Balderramo 5 , Hernan 

Coseano 5 , Viviana Re 3 ; 1 University of MInnesota, Minneapolis, 

MN; 2 Área de Virología y Biología Molecular, LACE Laboratorios, 

Cordoba, Argentina; 3 Instituto de Virología “Dr.J.M.Vanella”, Facultad 

de Ciencias Médicas, Universidad Nacional de Córdoba, 

Cordoba, Argentina; 4 Hospital Rawson, Cordoba, Argentina; 

5 Hospital Privado, Cordoba, Argentina 

Background: Hepatitis E virus (HEV) is a single-stranded RNA 

virus that can cause hepatitis in an epidemic fashion. In immunocompetent 

individuals, infection with HEV usually leads to 

silent seroconversion or to acute self-limited disease. Several 

reports suggest an increased rate of seroprevalence of HEV in 

immunosuppressed individuals, particularly those undergoing 

solid-organ transplantation. In this setting, HEV can develop 

into a chronic infection. The data is less clear in patients 

infected with human immunodeficiency virus (HIV). Moreover, 

information about prevalence of HEV in immunocopromised 

subjects outside of Europe or North America is scarce. In this 

study we addressed the seroprevalence of HEV in immunocompromised 

subjects in Argentina and associated risk factors. 

Methods: We performed third generation enzyme immunoassay 

for determination of IgG specific antibodies against HEV 

in 95 subjects infected with HIV, 81 subjects on hemodialysis 

(HD) and 58 solid-organ transplant recipients. On those samples 

that were positive for HEV IgG, further assessment of HEV 

IgM levels. HEV PCR was performed in all samples. Subjects on 

HD and transplant recipients were evaluated regarding social 

habits and potential risk factors. Results were compared to 

433 HIV-negative, immunocompetent controls from our center. 

Results: In our entire HIV-positive group we found 8 of 

95 samples to be positive for HEV IgG (8.8%), compared to 

19 of 433 samples (4.4%) in the control group. Interestingly, 

in a subgroup of patients (N27) with low CD4 counts 18% 

were positive for HEV IgG (p=0.03, compared to controls). 

This group had a much lower CD4 count when compared to 

the general HIV cohort (median CD4 count of 43/mm3 vs 

543/mm3 respectively). Eight out of 81 subjects (9.8%) on HD 

and 5 of 58 (8.6%) of transplant recipients were positive for 

HEV IgG. Interestingly, half of HEV seropositive patients in the 

HD group had positive IgM for HEV. There was no association 

between HEV serostatus and consumption of pork, fish, potable 

water or history of blood transfusion. Only 1 sample showed 

a positive PCR for HEV, within the HIV group. This patient 

had recent history of vomiting and diarrhea and likely experienced 

acute HEV infection. Conclusions: Our study found an 

increased seroprevalence of HEV IgG in subjects infected with 

HIV, on HD and solid-organ transplant recipients in Argentina. 

However, the only significant difference compared to controls 

was on HIV-infected patients with low CD4 counts. 

Disclosures: 

The following authors have nothing to disclose: Jose Debes, Maribel Martinez 

Wassaf, Maia Belen Pisano, Leonardo Marianelli, Martin Lotto, Domingo Balderramo, 

Hernan Coseano, Viviana Re 

1753 

Oral Interferon-free Antiviral Regimens in Liver Transplant 

Recipients with Hepatitis C: a Real Life Experience 

Arun Mannem 2,1 , Shari S. Rogal 2 , Vivek Sharma 2 , Fadi Francis 2 , 

Thomas V. Cacciarelli 2 , Obaid S. Shaikh 2,1 ; 1 Medicine/Gastroenterology, 

University of Pittsburgh School of Medicine, Pittsburgh, 

PA; 2 Medicine/Gastroenterology, Veterans Affairs Pittsburgh 

Healthcare System, Pittsburgh, PA 

Purpose: Oral interferon-free regimens are highly effective for 

hepatitis C (HCV) in the immunocompetent population. However, 

their role in transplant recipients is not well established. 

This study aimed to determine the real life experience with 

such regimens in liver transplant recipients. Methods: Beginning 

in 2013, all patients evaluated for HCV at VA Pittsburgh 

Healthcare System were prospectively entered in a quality 

of care database. One hundred twenty five liver transplant 

recipients with HCV were identified; 25 were excluded [treatment 

with interferon based regimens (13), transplant at a 

different center (6), lack of complete data (4), primary graft 

failure (2), and treatment in a clinical trial (1)]. Data regarding 

demographics, prior post-transplant treatment, HCC and 

MELD status and treatment were collected. Results: Among 100 

patients studied, 96 received transplant during 2001-2014. 

Most patients were adult, white males and received tacrolimus 

based immunosuppression. Eighty-six (86%) had genotype 1, 

1 (1%) genotype 2, 12 (12%) genotype 3 and 1 (1%) genotype 

4. Among genotype 1 patients, 60/86 (70%) underwent 

treatment post-transplant; 26/60 (43%) received combination 

sofosbuvir/simeprevir (SOF/SIM), 5 (8%) ledipasvir/sofosbuvir 

(LED/SOF) x 12 weeks, 6 (10%) LED/SOF x 24 weeks, 

21 (35%) LED/SOF/ribavirin (RBV) x 12 weeks and 1 (2%) 

received LED/SOF/RBV x 24 weeks. Rapid virologic response 

rates at 4 weeks of treatment (RVR) follow: - SOF/SIM 16/26 

(62%), LED/SOF-12 weeks 2/5 (40%), LED/SOF-24 weeks 

3/6 (50%), LED/SOF/RBV-12 weeks 14/21 (67%) and LED/ 

SOF/RBV-24 weeks 0/1 (0%). The single genotype 2 patient 

was treated with SOF/RBV and response is awaited. Four genotype 

3 patients were treated- 1 (25%) received SOF/SIM, 

2 (50%) LED/SOF x 12 weeks and, 1 (25%) SOF/RBV. The 

single genotype 4 patient had treatment with LED/SOF/RBV x 

12 weeks and achieved RVR. The full treatment outcomes are 

awaited and will be reported at the meeting. Conclusion: Our 

study will shed light on real world treatment outcomes with the 

all oral interferon-free regimens in liver transplant recipients 

with HCV. It will help establish the tolerability, efficacy and 

duration of therapy in this special population. 

Disclosures: 

Shari S. Rogal - Grant/Research Support: Gilead Sciences


Obaid S. Shaikh - Grant/Research Support: Gilead Sciences, Shinongi Pharmaceuticals; 

Speaking and Teaching: Simply Speaking 

The following authors have nothing to disclose: Arun Mannem, Vivek Sharma, 

Fadi Francis, Thomas V. Cacciarelli 

1754 

Effect of simeprevir administration on dose-normalized 

concentration of calcineurin inhibitors in solid organ 

transplant recipients with hepatitis C infection 

Heather Johnson 1,2 , Maxwell A. Brown 2 , Alicia B. Lichvar 2 , 

Michael A. Dunn 1 , Kapil B. Chopra 1 ; 1 University of Pittsburgh, 

PIttsburgh, PA; 2 UPMC Presbyterian, Pittsburgh, PA 

Background Simeprevir (SMV) is a second generation NS3/4A 

protease inhibitor that lacks the immunomodulatory and other 

adverse effects of interferon, representing an attractive therapeutic 

alternative for the treatment of hepatitis C virus infection 

in solid organ transplant recipients. The effect of P-glycoprotein 

(P-gp) inhibition by SMV on serum concentrations of the calcineurin 

inhibitors (CNI) cyclosporine (CsA) and tacrolimus (TAC) 

is not well described in the literature. The purpose of this analysis 

was to determine the magnitude of effect of P-gp inhibition 

with SMV on CNI blood levels. Methods Liver transplant recipients 

(LTRs) on maintenance immunosuppression with a CNI 

who initiated antiviral treatment with SMV between 1/2014 

and 4/ 2015 were included in this analysis. SMV was administered 

along with the NS5B polymerase inhibitor sofosbuvir for 

recurrent Genotype 1 allograft HCV. Dose-normalized levels 

were calculated by dividing CNI trough levels by the CNI total 

daily dose in order to account for CNI dose changes related 

to changes in CNI trough levels over time. Dose-normalized 

CNI levels in patients receiving 12 weeks of SMV were compared 

using Friedman’s ANOVA. Median percent change in 

dose-normalized levels were compared between LTRs with low 

(0–2) and high (3–6) Metavir scores using the Mann-Whitney 

U test. Results Nineteen patients were identified for inclusion in 

this analysis, of whom 15 (78.9%) received 12 weeks of SMV 

and 4 (21.1%) received 24 weeks of SMV. The majority of 

patients were Caucasian (100.0%), male (63.2%), liver transplant 

recipients (100.0%) on maintenance immunosuppression 

with TAC (n=17) with an average age of 59.2 years (SD±6.7). 

There was no significant difference between dose-normalized 

levels of TAC at baseline and during HCV treatment. Three 

patients (15.8%) had an increase in CNI dose and 4 patients 

(21.1%) had a decrease in CNI dose following initiation of 

SMV. There was no significant difference in median percent 

change in dose-normalized TAC levels between groups with 

low and high Metavir scores. Conclusions Patients on maintenance 

immunosuppression with TAC who were treated with 12 

weeks of SMV did not experience significant changes in their 

dose-normalized TAC trough levels. More frequent monitoring 

of TAC trough levels does not appear to be warranted in transplant 

recipients initiating SMV therapy. 

Disclosures: 

The following authors have nothing to disclose: Heather Johnson, Maxwell A. 

Brown, Alicia B. Lichvar, Michael A. Dunn, Kapil B. Chopra 

1755 

Bioluminescence imaging of mesenchymal stem cells by 

over expression hepatocyte nuclear factor4α: tracking 

survival and biodistribution 

Pei-yi Xie, Xiaojun Hu, Xiaochun Meng, Hong Shan; Department of 

Radiology, The Third Affiliated Hospital of Sun Yat-sen University, 

Guangzhou, China 

Hepatocytes from human bone marrow-derived mesenchymal 

stem cells (hBM-MSCs) are expected to be a useful source 

for cell transplantation. However, relatively low efficiency of 

hepatic differentiation of hBM-MSCs remains unsolvable in clinical 

application. Hepatocyte nuclear factor 4alpha (HNF4α), 

a critical transcription factor, is essential for the entire process 

of liver development. The purpose of this study was to construct 

MSCs with over expression HNF4α and investigate their 

hepatic differentiation and therapeutic potential for treating 

Fulminant hepatic failure (FHF) rats, and track their survival 

and biodistribution after transplantation by bioluminescence 

imaging (BLI). HNF4α gene was transduced into hBM-MSCs by 

lentiviral vector (pLV/Final-puro-hHNF4α-hrGFP). HNF4α-transduced 

MSCs (E7-hHNF4α) and GFP-transduced MSCs cells 

(E7-hrGFP ) were labeled with pLENTi-CMV-luc2-mKate2 and 

analyzed for their hepatic functions such as measurements 

of albumin, urea, glucose, cytochrome P450 activity,Indocyanine 

green (ICG) uptake and release, and drug metabolization 

in vitro. FHF modals of Sprague-Dawley (SD) rats were 

established and divided into three groups: PBS, E7-hrGFP and 

E7-hHNF4α cells’ transplantation. After 2.0×10 6 cells transplantation, 

BLI was used to dynamically track the survival and 

biodistribution of implanted E7-hrGFP cells and E7-hHNF4α 

cells. The restoration of biological functions of the livers receiving 

transplantation was assessed via a variety of approaches 

such as mortality rate determination, serum biochemical analysis, 

and histological, immunohistochemical, and genetic analysis. 

In vitro, E7-hHNF4α cells showed mature hepatic functions 

including albumin secretion, urea production, glycogen storage, 

cytochrome P450 activity, ICG uptake and release and 

drug metabolization. They improved liver functions in vivo after 

transplantation into the D-galactosamine-injured rats’ liver as 

evidenced by the fact that AST, ALT, TBIL returned to normal 

levels in recipient FHF rats. The result of 30-day survival rates 

suggested that E7-hHNF4α cells’ transplantation via superior 

mesenteric vein (SMV) was able to significantly prolong the 

survival of FHF rats compared with the other two groups. BLI, 

histochemisty, and RT-PCR results confirmed the presence of 

transplanted E7-hrGFP cells and E7-hHNF4α cells in recipient 

rat livers. CONCLUSION: Our data revealed that E7-hHNF4α 

cells could not only differentiate into functional hepatocyte-like 

cells in vitro, but could also improve liver functions and prolong 

the survival time of FHF rats. And BLI is a useful tool to track the 

transplanted cells survival and biodistribution. 

Disclosures: 

The following authors have nothing to disclose: Pei-yi Xie, Xiaojun Hu, Xiaochun 

Meng, Hong Shan


1756 

Dual CCR2/CCR5 Antagonist Cenicriviroc Leads to 

Potent and Significant Reduction in Proinflammatory 

CCR2+ Monocyte Infiltration in Experimental Acute Liver 

Injury 

Oliver Krenkel 1 , Tobias Püngel 1 , Jana C. Mossanen 1 , Can Ergen 1 , 

Felix Heymann 1 , Eric Lefebvre 2 , Christian Trautwein 1 , Frank 

Tacke 1 ; 1 Medical Clinic III, University Hospital Aachen, Aachen, 

Germany; 2 Tobira Therapeutics Inc., San Francisco, CA 

Aim of the study: Acute liver failure (ALF) is a life-threatening 

condition with rapid deterioration of hepatic function and 

limited therapeutic options. In mouse models, liver injury by 

toxic agents like carbon tetrachloride (CCl4) or acetaminophen 

(APAP) leads to rapid pro-inflammatory monocyte infiltration 

into the liver via the CCR2–CCL2 (a.k.a. MCP-1) chemokine 

pathway. Cenicriviroc (CVC) is an oral, once-daily CCR2/ 

CCR5 antagonist currently evaluated in a Phase-2b clinical trial 

in adults with NASH and liver fibrosis. We therefore evaluated 

CVC for inhibiting monocyte infiltration in CCl4 and APAP-induced 

acute liver injury in vivo. Methods: C57BL/6J (WT) and 

CCR2-deficient mice were subjected to ALF either by CCl4 

(0.6 ml/kg IP) or APAP (250 mg/kg IV). In both models, mice 

received either CVC (100 mg/kg) or vehicle by oral gavage. 

Liver injury and immune cell phenotypes were analyzed. For 

mechanistic studies, monocyte-derived macrophage subsets 

and resident macrophages (Kupffer cells) were sorted by FACS 

from injured livers and subjected to array-based Nanostring 

gene expression analysis. Results: Both CCl4 and APAP led 

to a rapid and massive accumulation of Ly6C+, monocyte-derived 

macrophages in injured livers, dependent on the chemokine 

receptor CCR2. Oral administration of CVC significantly 

reduced pro-inflammatory Ly6C+ monocytes in blood (p


1758 

Arcuate artery resistive index predicts Acute-on-Chronic 

Liver Failure 

Pablo Solis-Munoz 2,1 , Christopher Willars 2 , Julia Wendon 2 , 

Georg Auzinger 2 , Michael A. Heneghan 2 , María De la Flor-Robledo 

3 , José A. Solís-Herruzo 1 ; 1 Research Center, Hospital “12 de 

Octubre”, Madrid, Spain; 2 Institute of Liver Studies, King’s College 

Hospital, London, United Kingdom; 3 University Hospital Severo 

Ochoa,, Madrid, Spain 

Patients with acutely decompensated (AD) liver cirrhosis are 

at risk for developing acute-on-chronic liver failure (ACLF) syndrome. 

This syndrome is associated with a high sort-term mortality 

rate. The aim of our study was to identify reliable and 

early predictors of developing ACLF in cirrhotic patients with 

AD. Methods: We assessed 84 cirrhotic patients admitted for 

AD without ACLF on admission. We performed routine blood 

testing and detailed ultrasound Doppler studies of systemic 

arteries and mayor abdominal veins and arteries. We also 

calculated liver and ICU predictive scores. The area under 

the ROC curve (AUROC) was calculated for all variables that 

were significantly different between patients who developed 

ACLF and those who did not. Sensitivity, specificity, positive 

and negative predictive values, and diagnostic accuracy for 

the prediction of the short-term ACLF development were determined. 

Results: Of the 84 patients, 23 developed ACLF and 61 

did not develop this complication. In the univariate analysis, 

serum levels of urea, creatinine, and bilirubin, prothrombin 

time, MELD score, portal vein, femoral and poplitea artery flow 

velocity, and renal and arcuate artery resistive indices (RI) had 

predictive value for short-term development of ACLF. However, 

only arcuate artery RI had independent predictive value in the 

multivariate analysis. The AUROC value for RI of the arcuate 

arteries was 0.9971. Conclusions: On the first day of admission, 

ultrasound measurement of the RI of the arcuate arteries 

recognizes with a high degree of certainty cirrhotic patients 

admitted because of AD who will develop ACLF. 

Disclosures: 

Julia Wendon - Consulting: Pulsion, Excalenz 

The following authors have nothing to disclose: Pablo Solis-Munoz, Christopher 

Willars, Georg Auzinger, Michael A. Heneghan, María De la Flor-Robledo, José 

A. Solís-Herruzo 

1759 

Fulminant Hepatitis B Acute Liver Failure due to immunosuppressant 

or chemo-therapy: A multicenter retrospective 

cohort analysis 

Filipe S. Cardoso 1 , Michelle Gottfried 3 , K. Rajender Reddy 2 , A. 

James Hanje 4 , Daniel Ganger 5 , William M. Lee 6 , Constantine J. 

Karvellas 1 ; 1 Hepatology/Critical Care Medicine, University of 

Alberta, Edmonton, AB, Canada; 2 University of Pennsylvania, Philadelphia, 

PA; 3 Medical University of South Carolina, Charlston, 

SC; 4 The Ohio State University, Columbus, OH; 5 Northwestern 

University, Chicago, IL; 6 University of Texas Southwestern, Dallas, 

TX 

Background/aims: Acute Liver Failure (ALF) due to reactivation 

of latent hepatitis B (HBVr) due to chemotherapy or immunosuppression 

(CTI) can be fatal and yet is preventable in many 

cases. Aims: To determine the frequency of HBV-ALF due to 

reactivation from CTI within the US Acute Liver Failure Study 

Group (ALFSG) registry and compare patient survival and complication 

rates to non-CTI HBV-ALF controls. Methods: Of a total 

of 149 patients with HBV induced ALF enrolled in the US ALFSG 

registry between 01/1998 and 04/2015, 26 (17%) developed 

HBV-ALF due to CTI and 123 (83%) did not (controls). 

Multivariable logistic regression was performed to determine 

predictors of 21-day spontaneous survival in HBV ALF, specifically 

examining the impact of HBVr ALF due to CTI. Results: 

Of the 26 CTI patients, 9 received corticosteroids, 15 cytotoxic 

chemotherapy (breast, lymphoma, testicular, leukemia, 

myeloma), 2 received rituximab and 1 received gemtuzumab. 

There were no differences between these groups in proportions 

of patients with coma grade 3 or 4 (67% vs. 63%, p=0.77), 

requirement for mechanical ventilation (32% vs. 46%, p=0.19) 

or renal replacement therapy (18% vs. 15%, p=0.70). Of CTI 

HBV-ALF patients, 10 (38%) were listed for liver transplant (LT) 

(vs. 46% controls, p=0.70) and 6 (26%) received LT (vs. 34% 

controls, p=0.37). Of these 6 LT patients, 2 died post-LT (survival 

post-LT 67% vs. 81% in controls, p=0.59). Overall unadjusted 

21-day survival was significantly lower (39% vs. 61%, 

p=0.015) in CTI HBVr- ALF patients but spontaneous survival 

was similar (25% vs. 35%, p=0.24) when compared with controls. 

Using multivariable logistic regression to examine clinical 

factors associated with 21-day spontaneous survival in HBV 

ALF patients, MELD (Odds Ratio 0.87 per increment; 95% CI 

0.80-0.95, p=0.0013) and maximum hepatic coma grade 

> 2 (OR 0.13, 95% CI 0.04-0.41, p=0.0005) were significant. 

There was a non-significant trend towards worse 21-day 

adjusted spontaneous survival in CTI HBV ALF patients (OR 

0.16, 0.02-1.54, p=0.11). The model performed well (c-statistic 

0.87). Summary/Conclusions: HBV ALF due to CTI has 

poor spontaneous survival (25%) and overall poorer outcomes, 

even after adjusting for MELD and Hepatic coma grade (>2). 

Guidelines from specialties that prescribe chemotherapy/immunosuppressant 

therapies need to more explicitly detail testing in 

who, how testing for those at risk should be performed, timing 

and duration of therapy, and surveillance for those at risk not 

on therapy. In this population of HBV-ALF patients where liver 

transplant may not be offered, prevention of reactivation is 

critical given the significant mortality observed. 

Disclosures: 









Constantine J. Karvellas - Grant/Research Support: Merck; Speaking and Teaching: 

Gambro 

The following authors have nothing to disclose: Filipe S. Cardoso, Michelle 

Gottfried 

1760 

Mesenchymal stem cell using scaffold treatment 

increased survival on acute liver failure model 

Yong Kyun Cho 1 , Dae Won Jun 2 , Eun Chul Jang 3 , Seung Min 

Lee 3 , Joo Hee Kwak 2 ; 1 Department of Internal Medicine, Kangbuk 

Samsung Hospital, Sungkyunkwan University, School of Medicine, 

Seoul, Korea (the Republic of); 2 Department of Internal Medicine, 

Hanyang University College of Medicine, Seoul, Korea (the Republic 

of); 3 Department of Occupational and Environmental Medicine, 

Soonchunhyang University Cheonan Hospital, Cheonan, Korea 


Background: Recently, the effects of mesenchymal stem cells 

(MSCs) have been extensively evaluated in acute liver failure 

model. However, the most commonly used methods are used 

injecting MSCs through a vessel, which has a major drawback 

of homing cells in other organs. The aim of current study was 

to evaluate the efficacy and degree of cell homing of scaffold 

loaded with MSCs on acute liver failure model. Methods: Acute


liver failure was induced in C57BL/6J mice using thioacetamide 

(TAA) (200mg/kg, i.p) once a day for 2 days. In phase 

I, animals were divided in three groups: 1) TAA+saline via tail 

vein; 2) TAA+MSCs via tail vein; 3) TAA+Scaffold loaded with 

MSCs to evaluate the mortality and changes in liver function. 

PLGA Poly (lactic-co-glycolic acid) scaffold alone, and loaded 

with 1x10 6 human MSCs were implanted on dorsum of animals. 

In phase II, animals were divided into three groups: 1) 

TAA; 2) TAA+Scaffold; 3) TAA+Scaffold loaded with MSCs 

to evaluate liver function and mortality. Animals were sacrificed 

3 days after implantation, serum and liver samples were 

collected. Liver tissue immunohistochemistry for proliferation 

marker (Ki67) was performed. Results: In phase I, the mortality 

rate of TAA+Scaffold group was 36% as compared to 

TAA+PBS tail vein (66%) and TAA+Scaffold only groups (54%). 

In phase II, TAA+Scaffold loaded with MSCs group had only 

11.8% mortality as compared to TAA (30.4%) and TAA+Scafold 

only (52.6%) (p=0.014) groups. In phase II, serum AST 

levels of TAA+Scaffold loaded with MSCs was lower than other 

two groups, while serum ALT levels of TAA+Scaffold group 

were the lowest. Furthermore, TAA+Scaffold loaded with MSCs 

group showed increased liver tissue staining for Ki67. Later, we 

performed PCR using human specific primer, to evaluate homing 

of MSCs in mouse liver. There was no detection of human 

cells residing in mouse liver. Conclusion: Scaffold loaded MSCs 

showed protective effects via paracrine signaling on acute liver 

failure model. 

Disclosures: 

The following authors have nothing to disclose: Yong Kyun Cho, Dae Won Jun, 

Eun Chul Jang, Seung Min Lee, Joo Hee Kwak 

1761 

Indeterminate Etiology of Acute Liver Failure: Fact or 

Fiction? 

Jody A. Rule 1 , Jorge Rakela 2 , Norman L. Sussman 3 , Nathan M. 

Bass 4 , Holly Tillman 5 , Jaime L. Speiser 5 , R. Todd Stravitz 6 , Ryan 

Taylor 7 , William M. Lee 1 , Daniel Ganger 8 ; 1 Internal Medicine, 

University of Texas Southwestern Medical Center, Dallas, TX; 2 Gastroenterology 

and Hepatology, Mayo Clinic Hospital, Phoenix, 

AZ; 3 Division of Abdominal Transplantation, Baylor College of 

Medicine, Houston, TX; 4 Department of Medicine, University of 

California San Francisco, San Francisco, CA; 5 Data Coordination 

Unit, Department of Public Health Sciences, Medical University 

of South Carolina, Charleston, SC; 6 Internal Medicine, Virginia 

Commonwealth University, Richmond, VA; 7 Gastroenterology and 

Hepatology, University of Kansas Medical Center, Kansas City, 

KS; 8 Gastroenterology and Hepatology, Northwestern Memorial 

Hospital, Chicago, IL 

Background: Approximately 11% of acute liver failure or 

acute liver injury (ALF/ALI) cases are initially listed as “indeterminate” 

by attending hepatologists at academic transplant 

centers; this has been attributed in the past to an unidentified 

virus or toxic injury. Although nearly 20% of these cases 

demonstrate a positive acetaminophen (APAP)-adduct assay 

(Hepatology 2011;53:567), the remainder still are considered 

indeterminate. Methods: We examined 302 indeterminate 

cases from 2713 consecutively enrolled patients in the 

ALFSG Registry using pre-defined characteristics developed by 

an expert committee, to identify each etiology as either Highly 

Likely/Definite (HL/D: >75% certainty) or Probable (>50% certainty). 

Examples: Criteria for autoimmune ALF (AI-ALF): calculated 

globulins elevated >ULN or serum IgG >ULN; ANA, 

ASMA or LKMA-positive titer >1:40; biopsy features: plasma 

cells, centrilobular necrosis or interface hepatitis. Thus, Highly 

Likely/Definite (HL/D) AI-ALF: All 3 criteria met, Probable AIH: 

Any 2 of 3 met. For APAP, HL/D: Positive APAP-adduct assay 

or history of APAP ingestion or APAP level (or both) and AST 

or ALT >1000/Bili 1 etiology. Among the remaining 

86 cases, 51 had at least 1 primary diagnostic test missing 

(APAP-adducts, ANA, aHAVIgM, HBsAg, aHBcIgM, or aHCV). 

Only 35 cases had a complete set of medical history and laboratory 

tests, and no etiology defined, fulfilling our diagnosis of 

“indeterminate” ALF. Among patients initially diagnosed with 

indeterminate ALF or ALI, 216 (72%) could be assigned to a 

single defined etiology with additional testingand standardized 

criteria. Dual etiology (or uncertainty with two possibilities) 

will need further adjudication. Incomplete history or diagnostic 

testing precluded assignment of etiology in 17%. Conclusions: 

Only 11.5% of “indeterminates”, 1.2% of the overall ALF registry, 

fulfilled stringent criteria for “indeterminate etiology” (data 

complete, no cause discernible). With a defined/adjudicated 

approach to causality, true “indeterminate” ALF is much less 

common than previously estimated. 

Disclosures: 

Norman L. Sussman - Advisory Committees or Review Panels: BMS, Merck; 

Board Membership: HepaHope; Grant/Research Support: AbbVie, BMS, Biotest, 

Conatus, Esai, Genfit, Gilead, Hyperion, Immuron, Intercept, Lumena, Merck, 

Novartis, Ocera; Speaking and Teaching: AbbVie, Gilead, Jannsen 

Nathan M. Bass - Advisory Committees or Review Panels: Quest Diagnostics 





The following authors have nothing to disclose: Jody A. Rule, Jorge Rakela, Holly 

Tillman, Jaime L. Speiser, R. Todd Stravitz, Ryan Taylor 

1762 

High CXCR-1 and CXCR-2 Expressing Neutrophils 

Induce Early Hepatocyte Death and promote Liver Injury 

in Acute-on-Chronic Liver Failure 

Arshi Khanam 1 , Nirupma Trehanpati 1 , Peggy Riese 2 , Archana Rastogi 

1 , Carlos A. Guzman 2 , Shiv K. Sarin 1 ; 1 research, Institute of 

liver and biliary sciences, New Delhi, India; 2 Helmholtz Centre for 

Infection Research, Braunschweig, Germany 

Background and aims: Acute-on-chronic liver failure (ACLF) is a 

serious and often fatal liver disease. Mechanisms of liver injury 

and development of sepsis in these patients are unclear. Neutrophils 

contribute to disease pathogenesis in acute liver failure 

and serve as a biomarker of organ dysfunction. We investigated 

the mechanisms of CXCR-1 and CXCR-2 expressing 

neutrophils mediated hepatic injury in ACLF patients (alcohol 

and hepatitis B related) and compared with chronic hepatitis 

B (CHB) and healthy controls (HC). Methods: CXCR-1 and 

CXCR-2 expressions on neutrophils were measured by flowcytometry, 

immunohistochemistry and RT-PCR. In vitro co-culture 

assays were performed to determine the mechanisms of hepatic 

injury and cell death. Mechanism of contact dependant cell 

death was investigated by co-culture of neutrophils with HepG2 

and HepG2.2.15 cells. Contact independent cell death was 

checked by culture of HepG2 and HepG2.2.15 cells with LPS 

stimulated neutrophil’s supernatant. In patients, cell death was 

determined by intrahepatic caspase-3 (apoptosis) and recep-


tor-interacting-protein kinase 3 (RIP-3; necrosis marker) expressions. 

To confirm the involvement of CXCR-1 and CXCR-2 in 

hepatocyte death, CXCR-1 and CXCR-2 blockade assay was 

done using SCH527123 antagonist. Results: High CXCR-1 

and CXCR2 expressing neutrophils were markedly increased in 

ACLF patients (P


CK19, and γH2AX and p21 expression verified DNA damage 

and related perturbations with incomplete liver regeneration. 

Phosphoprotein analysis of differences in injured versus healthy 

livers using normalization with housekeeping gene products 

and >2-fold up- or downregulated phospho- versus total proteins 

indicated abnormalities in ATM signaling pathways 

(e.g., ATM, Chek2, CDKN1a, CDNK1b, E2F1, Gadd45, 

Rad51, p53, p73), cell cycle regulators (e.g.,CCNB1, CCNC, 

CCND3, CCNE1, CDC25c, Cdk1, Cdk2, Cdk, Cdk7) besides 

cell stress, inflammation, etc. IPA mapping of these and other 

proteins indicated dysregulations in DNA damage/ repair and 

G0/G1 and G2/M checkpoint controls. In cultured HuH-7 

cells, APAP cytotoxicity reproduced double-strand DNA breaks, 

dysregulation in ATM signaling and G0/G1 and G2/M checkpoint 

restrictions. Conclusions: APAP-induced ALF in human livers 

was characterized by replicative stress and arrest of cells in 

G0/G1 due to DNA damage with dysregulated ATM signaling 

and cell cycle checkpoint controls. These molecular alterations 

offer therapeutic targets for drug development with or without 

cell therapy to restore liver regeneration in ALF. 

Disclosures: 

The following authors have nothing to disclose: Preeti Viswanathan, Yogeshwar 

Sharma, Sriram Bandi, Sanjeev Gupta 

The baseline characteristics of all paracenteses episodes before 

and after propensity score matching 

Disclosures: 

The following authors have nothing to disclose: Su Lin 

1765 

Lower fibrinogen level predicts hemorrhagic complications 

following abdominal paracentesis in patients 

with acute-on-chronic liver failure: A propensity score 

analysis 

Su Lin; Liver Research Center of the First Affiliated Hospital of 

Fujian Medical University, Fuzhou, China 

Object Patients with acute on chronic liver failure (ACLF) usually 

present severe coagulopathy. Abdominal paracentesis is 

often performed in those patients. The aim of this study was to 

analyze the prevalence of hemorrhagic events after abdominal 

paracentesis and the predictive factors of it in ACLF populations. 

Methods Patients who were diagnosed as ACLF from 

1 January 2010 to 31 December 2014 were enrolled. Those 

who had any signs of ascites underwent routine paracentesis 

upon and after admission. A propensity score matching 

analysis (greedy nearest neighbor matching) was used to 

select matched cases from overall non-hemorrhagic group as 

a control group. Hemorrhagic complications and the risk factors 

were examined using bilinear logistic regression analysis. 

Results A total of 602 abdominal paracenteses were carried 

out on 218 ACLF patients within a 5-year period. A total of 

18 (2.99%) hemorrhagic complications were identified, with 

4 cases of abdominal wall hematomas and 14 cases of intraperitoneal 

hemorrhage. We finally matched 18 cases with 

bleeding events to 72 unique cases without bleeding cases. 

The patients with hemorrhagic events had significantly lower 

fibrinogen levels and higher PT level than non-hemorrhagic 

ones. Bilinear regression revealed that lower fibrinogen level 

could independently predict hemorrhagic complication (OR: 

0.128, 95% CI 0.023-0.697, p = 0.017). The best cut-off 

value of fibrinogen level f>or predicting bleeding events>was 0.70 g/L, 

with the sensitivity of 76.4% and specificity of 80.0%. The area 

under curve was 0.733 (95% CI 0.604-0.862, P =0.002). 

Conclusion Severe hemorrhagic complications happened in 

2.99% of all paracenteses performed for ACLF patients. Low 

fibrinogen level (≤0.70g/L) is an independent predictor. 

1766 

Does weight loss surgery predispose to acetaminophen-related 

acute liver failure? 

Shannan Tujios 2 , Michelle Gottfried 1 , Valerie L. Durkalski 1 , William 

M. Lee 2 ; 1 Medical University of South Carolina, Charleston, 

SC; 2 University of Texas Southwestern, Dallas, TX 

Background: Obesity and bariatric surgery are increasingly 

common with nearly 180,000 Americans undergoing weight 

loss surgery (WLS) annually; an estimated 4.5 million individuals 

in the United States have had bariatric surgery to date. A 

previous study (J Clin Gastro 2014 epub) suggested that prior 

WLS was over-represented among patients with acetaminophen 

(APAP) related acute liver injury or acute liver failure 

(ALI/ALF). Aims: To describe the presentation, etiology and 

outcome of ALI/ALF patients with prior WLS, compared to a 

large group of ALF patients with acetaminophen (APAP) overdose 

without prior WLS. Methods: 57 patients with prior WLS 

enrolled in the US ALFSG registry between 01/1998 and 

04/2015 were reviewed and compared to 1234 remaining 

non-WLS APAP patients. Results: During the early study era 

(1998-2009), 1.5% of all cases (N=1678) reported prior WLS, 

while 3.2% of all cases since (2010-2015) (N=997) had prior 

WLS. Those with prior WLS were largely female (96.5%), and 

white (84.2%), and most demonstrated advanced coma grades 

(3-4; 75%) during hospitalization; 43 (75.4%) were APAP-related, 

3 (5.3%) due to idiosyncratic drug injury and 4 (7%) 

indeterminate. Nearly 25% of WLS APAP patients were listed 

for liver transplant with 9 patients (64% of those) receiving a 

graft; 21 day spontaneous survival was 58.5%, compared to 

unadjusted overall survival of 61.4%. Comparing the cohort 

of 43 APAP patients with prior WLS to the remaining APAP 

patients, there were no differences in age, race or reported 

alcohol use. The WLS group was more often female (95.4% vs 

73.7%, p=0.0014), and was more anemic (median hemoglobin 

9.5 gm/dL vs 11.2 gm/dL p=


coma grade (74.3% vs 63.5%), percent waitlisted for transplant 

(23.3% vs 20.3%), fraction transplanted (14% vs 7.3%, 

p=0.2564) or 21 day spontaneous survival (64.1% vs 70.2%), 

overall unadjusted survival (65.1% vs 65.2%). Summary/Conclusions: 

The fraction of patients with ALF who have had prior 

WLS has been increasing in recent years as WLS becomes 

more commonplace. When ALF occurs in a patient with prior 

WLS, APAP toxicity is the most likely but not the only diagnosis 

observed. Bariatric surgery may predispose women to severe 

unintentional liver injury at relatively low APAP doses although 

outcomes appear similar with and without WLS. 

Disclosures: 



The following authors have nothing to disclose: Shannan Tujios, Michelle 

Gottfried, Valerie L. Durkalski 

1767 

Dual Role of Epidermal Growth Factor Receptor (EGFR) 

in Liver Injury and Regeneration after Acetaminophen 

Overdose in Mice 

Bharat Bhushan, Michael Manley, Mitchell R. McGill, Hartmut 

Jaeschke, Udayan Apte; Pharmacology, Toxicology & Therapeutics, 

University of Kansas Medical Center, Kansas City, KS 

Acetaminophen (APAP) overdose is the major cause of acute 

liver failure in the US with very limited treatment options. 

Compensatory liver regeneration following APAP toxicity is a 

critical determinant in final recovery and survival. Epidermal 

Growth Factor Receptor (EGFR) signaling is known to play an 

important role in liver regeneration after partial hepatectomy. 

However, role of EGFR signaling in APAP toxicity and accompanying 

compensatory regeneration is completely unknown. 

We investigated the role of EFGR in APAP-induced liver injury 

and subsequent liver regeneration using pharmacological inhibition 

of EFGR signaling in mice by treatment with Canertinib, 

a water soluble irreversible EGFR inhibitor. EGFR was remarkably 

activated as early as 30 minutes after APAP treatment in 

a dose-dependent manner and remained activated up to 24 hr 

after APAP treatment. Treatment with Canertinib 1 hr post-APAP 

caused robust inhibition of EGFR activation resulting in remarkable 

attenuation of APAP-induced liver injury. Canertinib treatment 

did not alter APAP-protein adduct formation indicating 

APAP metabolism was not altered by EGFR inhibitor treatment. 

Interestingly, APAP mediated JNK activation (a key mediator 

of APAP toxicity) was not altered by EGFR inhibitor treatment. 

In order to study role of EGFR in liver regeneration after APAP 

overdose, independent of injury, EGFR inhibitor was administered 

12 hr after APAP treatment, which resulted in remarkable 

impairment of liver regeneration response without any alteration 

of injury. Decreased liver regeneration was associated 

with decreased induction of cyclin D1 and decreased phosphorylation 

of Rb protein. In conclusion, our study revealed, 

for the first time, that EGFR receptor plays an important role in 

development of APAP injury but also has a direct role in stimulation 

of liver regeneration after APAP overdose. These data 

support the hypothesis that EGFR signaling plays dual role in 

APAP-induced hepatocyte death and proliferation in liver. 

Disclosures: 

The following authors have nothing to disclose: Bharat Bhushan, Michael Manley, 

Mitchell R. McGill, Hartmut Jaeschke, Udayan Apte 

1768 

Heat Stroke Leading to Acute Liver Failure: Case Series 

from the Acute Liver Failure Study Group 

Brian C. Davis 1 , Holly Tillman 2 , Robert J. Fontana 3 , K. Rajender 

Reddy 4 , Raymond T. Chung 5 , Brendan M. McGuire 6 , R. Todd Stravitz 

7 , William M. Lee 1 ; 1 Division of Digestive and Liver Diseases, 

UT Southwestern Medical Center, Dallas, TX; 2 Department of Public 

Health Sciences, Medical University of South Carolina, Charleston, 

SC; 3 Division of Gastroenterology, University of Michigan Health 

System, Ann Arbor, MI; 4 Division of Gastroenterology, University 

of Pennsylvania Perelman School of Medicine, Philadelphia, PA; 

5 Department of Gastroenterology, Massachusetts General Hospital, 

Boston, MA; 6 Division of Gastroenterology and Hepatology, 

UAB School of Medicine, Birmingham, AL; 7 Division of Gastroenterology, 

VCU Medical Center, Richmond, VA 

Introduction: In the United States, a small percentage of cases 

of acute liver injury (ALI) and failure (ALF) have been associated 

with heat stroke, defined as elevated body temperature 

≥40°C with central nervous system dysfunction. This may occur 

as classical heat stroke (CHS), defined as exposure to high 

environmental temperatures, or exertional heat stroke (EHS), 

which occurs after strenuous activity. The aim of this study was 

to describe cases of ALI or ALF caused by heat stroke in a large 

ALF registry. Methods: Amongst 2,675 consecutive ALI and ALF 

subjects enrolled between January 1998 and April 1, 2015, 

there were 8 subjects with heat stroke, classified as having 

CHS or EHS. The clinical, laboratory, and outcome data were 

then compared. Results: Five patients had ALF and 3 patients 

had ALI. Six patients had EHS, one with CHS, and one was 

unclassified. Seven patients developed acute renal failure, 8 

had lactic acidosis, and 6 had rhabdomyolysis. Six patients 

underwent cooling treatments, 3 received N-acetyl cysteine 

(NAC), 3 required mechanical ventilation, 3 required renal 

replacement therapy, 1 underwent liver transplantation, and 2 

patients died—both within a day of presentation. All but one 

case occurred between June and August (May). No long-term 

sequelae were observed in survivors. Conclusions: In the US, 

heat stroke accounts for



Merck, BMS 




The following authors have nothing to disclose: Brian C. Davis, Holly Tillman, R. 

Todd Stravitz 

1769 

Recovery of severe critically ill Acute Liver Failure (ALF) 

patients is associated with significantly better survival 

than patients undergoing liver transplantation 

Antonella Putignano 1 , Francesco Figorilli 1 , Banwari Agarwal 2 , 

Rajiv Jalan 1 ; 1 Liver failure group, UCL Institute for Liver and Digestive 

Health, UCL Medical School, Royal Free Campus,, London, 

United Kingdom; 2 Intensive Care Unit, Royal Free Hospital, Royal 

Free Hampstead NHS Trust, University College London, London, 


Introduction: In patients with ALF fulfilling poor prognostic criteria, 

mortality rates without liver transplantation (LT) are high. In 

a recent study (1), unexpectedly, even the mortality of patients 

with ALF who recovered spontaneously was reported to be 

high. Due to the multicenter nature of the study, long-term follow 

up data for many patients was not defined. In this single 

center study, we aimed to confirm or refute this observation in 

ALF patients admitted to a single intensive care unit. Methods: 

Consecutive ALF patients admitted between 1990-2014 were 

considered and the 90-day survivors (Early Survivors) included 

to evaluate the 3-year outcome. According to ALF etiology and 

management, 4-cohorts were identified: Paracetamol Overdose 

(POD) Transplanted, POD Not-Transplanted, Non-POD 

Transplanted and Non-POD Not-Transplanted. Chi Square or 

Fisher test were used to compare outcome between cohorts 

(p< .05) and Kaplan Meier analysis to estimate their cumulative 

survival. Predictors of 3-year mortality were modeled 

with Cox Regression analyses. Causes of mortality were also 

identified. Results: 200 ALF patients were admitted to intensive 

care unit, and 124 were Early Survivors. 112/124 reached 

the 3-year follow up and were included in this study (Female 

77, age 35,4 + 11.7, POD 58). 13/112 POD Transplanted 

(11.6%), 45/112 POD Not-Transplanted (40.2%), 30/112 

Non-POD Transplanted (26.8%) and 24/112 Non-POD 

Not-Transplanted (21.4%) were identified (p


1771 

Cathepsin L-deficiency enhances liver regeneration after 

partial hepatectomy 

Toshifumi Sato, Shunhei Yamashina, Kousuke Izumi, Hirofumi 

Fukushima, Yoshihiro Inami, Tomonori Aoyama, Akira Uchiyama, 

Kazuyoshi Kon, Kenichi Ikejima, Sumio Watanabe; Department of 

Gastroenterology, Juntendo University School of Medicine, Tokyo, 

Japan 

Background Cathepsin L (CTSL), known as a highly potent 

endoprotease of the cysteine cathepsin family, plays a pivotal 

role on lysosomal protein degradation. It was reported that 

the activity of collagenolytic cathepsins including CTSL was 

enhanced in the CCl 4 

-induced fibrotic liver. On the other hand, 

previous investigations revealed lysosomal proteolysis is suppressed 

accompanied by decreased biosynthesis of cathepsins 

in liver after partial hepatectomy. The role of decreased CTSL 

level in liver regeneration has been still unclear. We investigate 

to clarify if CTSL-deficiency affects liver regeneration after partial 

hepatectomy. Methods 70% of partial hepatectomy (PH) 

was performed in male CTSL-deficient (Ctsl-/-) mice and wildtype 

littermates (Ctsl +/+). Mice were sacrificed and wet weight 

of the whole remaining liver was measured after 2 and 5days. 

Restituted liver mass was expressed as percentage of the ratio 

of remaining liver divided by calculated-total pre-hepatectomy 

liver weight. Bromodeoxyuridine (BrdU)-immunostaining of liver 

sections was performed. Expression of cyclin D1 in the liver 

was evaluated by western blot analysis. Results Restituted liver 

mass at 5day after PH in Ctsl-/- mice was higher than wild-type 

mice significantly (p=0.008). BrdU incorporation into nucleai 

was observed in 5.16±0.90% of hepatocytes 48 hours after 

PH in wild-type mice; however, the rate of BrdU incorporation 

was significantly enhanced to 8.82±0.35% in Ctsl-/- mice. Protein 

level of CyclinD1 was increased in wild-type mice 48 hours 

after PH. Cathepsin-deficiency enhanced increase in cyclinD1 

expression after PH to about 3-fold of wild-type. Conclusion 

CTSL-deficiency accelerates regenerating response in the liver 

after PH. These findings suggested that the decreasing CTSL 

plays an important role on liver regeneration. Therefore, an 

increase in CTSL activity observed in CCl 4 

-induced fibrotic liver 

may attenuate liver regeneration. Taken together, CTSL might 

be a new therapeutic target on disorder of liver regeneration 

in hepatic cirrhosis. 

Disclosures: 

The following authors have nothing to disclose: Toshifumi Sato, Shunhei Yamashina, 

Kousuke Izumi, Hirofumi Fukushima, Yoshihiro Inami, Tomonori Aoyama, 

Akira Uchiyama, Kazuyoshi Kon, Kenichi Ikejima, Sumio Watanabe 

1772 

The Receptor Interacting Protein Kinase 3 is a critical 

early warning signs for onset process of acute-onchronic 

hepatitis B liver failure 

Yubao Zheng, Yurong Gu, Ke Wang, Min Zhang, Xiaohui Huang, 

Ziyu Lin, Liang Peng, Zhiliang Gao; Department of Infectious Diseases, 

The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, 

China 

Background and aims: Hepatitis B virus is a principal cause of 

Acute-on-chronic hepatitis B liver failure (ACHBLF) in the China. 

The pathophysiology of ACHBLF is incompletely understood. 

Recent findings suggest that the receptor interacting protein 

kinase 3 (RIP3) acts as a switch from apoptosis to necrosis. 

Thus, the aim of the current investigation was to determine 

if RIP3 is involved in onset process of ACHBLF or Hepatitis B 

virus-induced hepatocytel death. Methods: The expression of 

RIP3 and Tumor Necrosis Factor-α(TNF-α) were retrospectively 

investigated in liver tissues of 46 patients with ACHBLF and 78 

patients with chronic hepatitis B(CHB) at the China South Hepatology 

Center. Meanwhile, in vitro the apoptotic and necrotic 

levels were investigated in the HepG2.2.15 cell, the genetically 

modified RIP3-expressing HepG2.2.15 cells co-cultured with 

Natural killer(NK) cells or not. Result: Our results showed that 

the levels of RIP3 expressing in Liver tissues were significantly 

higher in ACHBLF than in CHB, as shown by immunohistochemical(IHC) 

and western blots. The TNF-α expression was similar 

to RIP3 that the expression levels of TNF-α increased more 

significantly in ACHBLF than in CHB. The rate of apoptosis in 

the genetically RIP3-over expressing HepG2.2.15 cells were 

significantly higher in HepG2.2.15 cells and the genetically 

RNA silencing RIP3 HepG2.2.15 cells co-cultured with NK 

cells in vitro. Result of cell experiments show that RIP3 protein 

effectively regulates the apoptosis and necrosis of HepG2.2.15 

co-cultured NK cells. Conclusion: Our study suggest that RIP3 

protein effectively regulates apoptotic of HepG2.2.15 co-cultured 

NK cells. And RIP3 is a critical early warning signs for 

onset process of ACHBLF. RIP3 may be a novel therapeutic 

target in the treatment of ACHBLF in the future. 

Figure:RIP3 expressing in Liver tissues and the rate of apoptosis in 

HepG2.2.15 cells co-cultured NK cells. 

Disclosures: 

The following authors have nothing to disclose: Yubao Zheng, Yurong Gu, Ke 

Wang, Min Zhang, Xiaohui Huang, Ziyu Lin, Liang Peng, Zhiliang Gao 

1773 

Nitric Oxide induced hepatic stellate cells apoptosis 

through autophagy inhibition in acute liver failure 

Yingli He 3 , Li Jin 1 , Jiuping Wang 2 , Xiaogang Zhang 5 , Jing Wang 1 , 

Jinfeng Liu 3 , Yuan Yang 3 , Taotao Yan 1 , Tianyan Chen 3 , Furong 

Cao 1 , Xiaoni Kou 4 , Yingren Zhao 3 ; 1 School of Medicine, Xi’an 

Jiaotong University, Xi’an, China; 2 Department of Infectious Diseases, 

Xijing Hospital, Fourth Military Medical University, Xi’an, 

China; 3 Department of Infectious Diseases, First Affiliated Hospital, 

School of Medicine, Xi’an Jiaotong University, Xi’an, China; 

4 Department of Hepatology, First Affiliated Hospital, Shaanxi 

University of Chinese Medicine, Xianyang, China; 5 Department 

of Hepatobiliary Surgery, First Affiliated Hospital, School of Medicine, 

Xi’an Jiaotong University, Xi’an, China 

Background & Aim Previously we have shown acute liver failure 

(ALF) accompanied by fibrosis, a process modulated by 

autophagy induction and activation of hepatic stellate cells 

(HSCs) and eventually favor for prognosis of ALF. Nitric oxide 

(NO), a multipotent mediator, has been shown to promote 

apoptosis in HSCs. However, the role NO played in ALF and


underlying mechanisms remain unclear. Methods ALF patients 

were recruited to investigate the correlation between plasma 

NO level and clinical feature. Plasma nitrite was measured 

using the Griess reaction. The expression of nitric oxide synthases 

(iNOS, eNOS and nNOS), HSCs activation (alpha- 

SMA), and autophagic activity (LC3) in liver were investigated 

using immunohistochemistry. HSCs were treated with NO 

donor, and then autophagy flux was investigated by immunoblotting 

and confocal microscopy. Apoptosis was quantified 

by characteristic DAPI stain nuclei. Results Firstly, plasma NO 

level is significantly increased in patients with ALF as compared 

with cirrhosis (53.60±19.74 μmol/L vs 19.40±9.03 

μmol/L, t=10.57, p


Disclosures: 



The following authors have nothing to disclose: Jody A. Rule, Linda S. Hynan, 

Nahid Attar, William J. Korzun 

Disclosures: 

William Bernal - Advisory Committees or Review Panels: Ocera Therapeutics ; 

Consulting: Vital Therapies Inc 

Julia Wendon - Consulting: Pulsion, Excalenz 

The following authors have nothing to disclose: Georg Auzinger, Robert Loveridge, 

Sameer Patel, Christopher Willars, Krishna Menon, Hector Vilca-Melendez 

1776 

Serial Procalcitonin Measurements in Acute Liver Failure 

Patients with Bacterial Infections 

Jody A. Rule 1 , Linda S. Hynan 1 , Nahid Attar 1 , William J. Korzun 

2 , William M. Lee 1 ; 1 Internal Medicine, University of Texas 

Southwestern Medical Center, Dallas, TX; 2 Allied Health, Virginia 

Commonwealth University, Richmond, VA 

Background: Multi-organ failure in ALF patients resembles 

severe sepsis and septic shock, making identification of bacterial 

infection can be difficult. Procalcitonin (PCT) has proven 

useful in sepsis in general to identify and predict response 

to infection. We hypothesized that serial PCT values in ALF 

patients might be predictive of outcome. Methods: Among ALF 

Study Group 47 patients with bacterial infection (positive culture 

on Day 1), we compared serial PCT levels in sera available 

at least for Days 1-4, using the ADVIA Centaur BRAHMS PCT 

assay, a sandwich chemi-luminescent immunoassay, comparing 

change in levels to outcome as spontaneous survival (SS) 

vs. death/transplantation (non-SS). Results: Elevated PCT levels 

were observed in all patients (median 2.31 ng/mL), falling rapidly 

in the SS group from median of 2.9 to 1.07 ng/mL by Day 

4, compared to the non-SS group that actually increased from 

1.42 to 2.04 ng/mL by Day 4. Both groups had an overall 

decrease by Day 7 of 70.3% (SS) and 49.3% (non-SS). Conclusions: 

High PCT levels are characteristic of most ALF subjects 

falling to


1778 

Early elevation in serum IFN-λ3 is responsible for progressive 

liver failure caused by acute infection of Hepatitis 

E virus genotype 4 

Jong-Hon Kang 1 , Kazumoto Murata 2 , Yoshiyasu Karino 3 , Takeshi 

Matsui 1 , Itaru Ozeki 3 , Kazuaki Takahashi 4 , Masahiro Arai 4 , 

Masashi Mizokami 2 ; 1 Center for Gastroenterology, Teine Keijinkai 

Hospital, Sapporo, Japan; 2 The Research Center for Hepatitis and 

Immunology, National Center for Global Health and Medicine, 

Tokyo, Japan; 3 Department of Hepatology, Sapporo Kosei General 

Hospital, Sapporo, Japan; 4 Department of Medical Sciences, 

Toshiba General Hospital, Tokyo, Japan 

Background / Aims: Acute sporadic infection of hepatitis E virus 

(HEV) has been emerging in non-endemic countries because of 

increasing clinical impacts. Hepatitis E virus (HEV) of genotype 

(Gt) 4 was showed to be more responsible for development of 

acute liver failure (ALF) compared with Gt 3 in our previous 

report (abstract ID; 745, AASLD 2014). However, the relationships 

between genotypic differences and disease severity have 

been obscure. Interferon (IFN)-λ3 has been recognized as one 

of markers for innate immune responses against viral infection, 

such as HCV infection. The aim of this study is to clarify the 

mechanism of disease progression in acute HEV of Gt 4 infection 

by the analysis of serum IFN-λ3 levels. Methods: Among 

98 patients with sporadic and autochthonous hepatitis E diagnosed 

in Hokkaido, Japan, a total of 43 patients in whom early 

sera were sampled during the day 2 to 14 after the onset were 

enrolled. Acute HEV infection was diagnosed upon the detection 

of HEV RNA by PCR or anti-HEV antibody (IgM) in sera by 

enzyme linked immune-sorbent assay (ELISA). HEV genotypes 

(Gt) were determined by comparison of a 326-nt sequence 

within ORF1 of HEV genome. IFN-λ3 levels were measured 

by chemiluminescence ELISA and HEV RNA were quantified 

by real time PCR for the same sera obtained from the patients. 

Acute liver failure (ALF) was defined to be a case with longer 

prothrombin time (INR>1.5). Results: Out of 43 patients with 

acute HEV (32 males, median age 56 years) thus collected, 15 

developed ALF, in which 2 presented hepatic coma. HEV Gt 

4 was determined in 27 patients, Gt 3 in 13, co-infection with 

Gt 3+4 in 1, and not determined in 2. IFN-λ3 level was 11.5 

(1.5-120.4) pg/ml at Day 4.5 (1-13) in median. In Gt 4 cases, 

median peak levels in AST and ALT were higher than those with 

Gt 3, respectively, and, as previously reported, Gt 4 infection 

was correlated with development of ALF in comparison with Gt 

3 (14/15 vs. 1/15, p=0.002, OR 16.2[95%CI, 1.9-140.1]). 

Serum IFN-λ3 levels were higher in Gt 4 cases than those in Gt 

3 cases (15.9 vs. 5.0 pg/ml, p=0.001), despite of no difference 

in sampling timing of sera between Gts. Among 27 cases 

with Gt 4 infection, IFN-λ3 levels were higher in ALF cases than 

in self-limited (26.3 vs. 12.8 pg/ml, p=0.046), and IFN-λ3 levels 

revealed correlation with HEV RNA titrations in sera in Gt 4 

(r=0.701, p=0.008). Conclusion: Our study may implicate the 

pathological roles of IFN-λ3 in acute hepatitis E especially ALF 

due to HEV Gt 4 infection, and serum IFN-λ3 would be a useful 

predictive marker for disease-severities. 

Disclosures: 


The following authors have nothing to disclose: Jong-Hon Kang, Kazumoto 

Murata, Takeshi Matsui, Itaru Ozeki, Kazuaki Takahashi, Masahiro Arai, 

Masashi Mizokami 

1779 

Expression of Krüppel-like factor 6 parallels induction of 

autophagy in acute liver injury 

Svenja Sydor 1 , Jan Best 1 , Paul P. Manka 1 , Sami Jafoui 1 , Martin 

Schlattjan 1 , Francisco Javier Cubero 2 , Thomas Schreiter 1 , Diana 

Vetter 3 , Andreas Paul 4 , Scott L. Friedman 5 , Guido Gerken 1 , Ali 

Canbay 1 , Lars Bechmann 1 ; 1 Department of Gastroenterology and 

Hepatology, University Hospital Essen, Essen, Germany; 2 Department 

of Medicine III, Gastroenterology, Metabolic Disease and 

Intensive Care, University Hospital RWTH Aachen, Aachen, Germany; 

3 Department of Surgery, University Hospital Zurich, Zurich, 

Switzerland; 4 Department of General- and Transplant -Surgery, 

University Hospital Essen, Essen, Germany; 5 Division of Liver Diseases, 

Mount Sinai School of Medicine, New York, NY 

Krüppel-like factor 6 (KLF6) is a ubiquitously expressed, multifunctional 

transcription factor and tumor suppressor gene regulating 

hepatocyte glucose and lipid homeostasis and KLF6 

down-regulation is associated with proliferation in hepatocellular 

cancer. Autophagy regulates turnover of long-lived or 

damaged organelles and proteins, which affects liver regeneration 

by maintaining intracellular energy production. With 

this study, we characterized the role of KLF6 in liver injury 

and regeneration and found significant correlations with the 

induction of autophagy. KLF6 expression was quantified in 

liver samples from 10 patients with acute liver failure (ALF) 

and 10 controls. Human liver preparations were treated with 

acetaminophen (APAP) up to 30 hours in an established ex 

vivo perfusion model. Furthermore, C57B/6 mice were sacrificed 

8h after APAP injection (6 mice/group) and HepG2 

cells were incubated with APAP for 24h. We characterized 

the role of KLF6 in liver regeneration in vivo in a model of partial 

hepatectomy (PHx) in wildtype (wt) or hepatocyte-specific 

KLF6 knockout mice (dKLF6) (6 mice/group). We performed 

an Affymetrix gene array (HT-MG4306) on liver samples of 

dKLF6 and wt mice at 12h after PHx. KLF6 expression was significantly 

induced in hepatocytes from ALF patients compared 

to controls. APAP perfusion of liver tissue significantly induced 

KLF6 expression and KLF6 was significantly upregulated in 

APAP treated mice. The gene array revealed significant alterations 

in autophagy related genes in dKLF6 mice compared to 

wt following PHx. Hepatocyte proliferation was significantly 

induced at early time points after PHx in dKLF6 mice compared 

to wt. Western blot analyses of whole liver tissue from dKLF6 

mice revealed reduced levels of LC3-II 72h after PHx. dKLF6 

mice had decreased expression levels of autophagy related 

molecules Atg7 and Beclin1 before and after PHx compared to 

wt mice. HepG2 cells and murine liver tissue showed increased 

LC3-II levels following APAP treatment. Here, we observed a 

consistent induction of hepatic KLF6 expression in various models 

of acute liver injury. Results from hepatocyte-specific KLF6 

knockout mice implicate a significant role for KLF6 in early 

hepatic regeneration. Furthermore, gene array data as well as 

studies on mRNA and protein expression in mice, humans and 

cell culture reveal a novel interaction between KLF6 and autophagy 

in acutely injured hepatocytes. The underlying mechanisms 

need further characterization. 

Disclosures: 

Jan Best - Speaking and Teaching: BTG 










Pharm; Stock Shareholder: Angion Biomedica, Intercept Pharma


The following authors have nothing to disclose: Svenja Sydor, Paul P. Manka, 

Sami Jafoui, Martin Schlattjan, Francisco Javier Cubero, Thomas Schreiter, Diana 

Vetter, Andreas Paul, Guido Gerken, Ali Canbay, Lars Bechmann 

1780 

Liver specific organ failure scoring systems, CLIF-OF and 

CLIF-ACLF are better than King’s College Hospital criteria 

in predicting mortality in patients with Acute Liver 

Failure (ALF) admitted to Intensive Care Unit 

Francesco Figorilli 2,1 , Antonella Putignano 2 , Banwari Agarwal 3 , 

Rajiv Jalan 2 ; 1 Universita’ di Cagliari, Cagliari, Italy; 2 UCL Medical 

School, Royal Free Campus, Liver failure group, UCL Institute for 

Liver and Digestive Health, London, United Kingdom; 3 Intensive 

Care Unit, Royal Free Hospital, Royal Free Hampstead NHS Trust, 

University College London, London, United Kingdom 

Background. ALF is a critical illness and a liver transplant (LT) 

is often the only life-saving treatment. King’s College Hospital 

criteria (KCH) are currently used to identify LT potential candidates. 

Given high mortality and the organ shortage, a more 

sensitive score is needed. In this study, nine different exiting 

scores (MELD, UKELD, iMELD, APACHE2, APACHE3, SOFA, 

KCH, CLIF-OF and CLIF-ACLF) were assessed at the admission 

to find the best predictor of mortality. Methods. Transplanted 

and non-transplanted patients were analysed separately and 

further stratified in base of aetiology (Paracetamol overdose 

(POD) or other causes (Non-POD)). Primary endpoint was the 

3-month mortality of any cause. For each score we assessed the 

area under the curve (AUC), the best cut-off value, sensitivity, 

specificity and positive predictive value (PPV). Cox analysis 

was used to identify the best predictor of 3-month mortality. 

Results. 200 patients admitted between 1990-2014 in our 

centre were included in this study (98 POD, female 65%, mean 

age 38.3±12,8). 70/200 received a LT (19 POD, 51 Non- 

POD), and 22 (31.4%) died within 3 months. In the Non LT 

cohort (79 POD and 51 NON POD), the mortality rate was 

41.5%. 90 patients fulfilled poor prognostic KCH criteria but 

did not receive a LT (12 were not listed for psychiatric problem, 

29 was considered too unwell for a LT and 31 spontaneously 

recovered). 18/90 were listed but 12 died on the waiting list 

and 6 improved. In Non-LT/Non-POD group CLIF-ACLF had 

the best AUC (0.799) with 73.9% sensitivity and 70.8% PPV; 

also it was the best mortality predictor in the multivariate analysis 

(p=0.001; HR 1.09; 95%CI 1.04-1.16). In Non-LT/POD 

CLIF-OF showed the second highest AUC (0.793) after SOFA 

(0.799) but with a higher sensitivity (93.5% vs 77.4%). In 

the univariate analysis CLIF-OF cut-off (12) had the highest 

Hazard Ratio (38.9). In Cox regression SOFA was the only 

significant in Non-LT/POD group (p


1782 

Histology predicts the need for liver transplantation in 

patients with acute severe autoimmune hepatitis 

Eleonora De Martin 1,2 , Audrey Coilly 1,2 , Mylene Sebagh 1,2 , Badr 

Serji 1 , Teresa Maria Antonini 1,2 , Eric Ballot 2,3 , Florent Artru 1 , 

Philippe Ichai 1,2 , Didier Samuel 1,3 , Jean-Charles Duclos-Vallee 1,2 ; 

1 Centre Hepato-Biliaire, AP-HP Hopital Paul Brousse, Villejuif, 

France; 2 Unit 1193, Inserm, Villejuif, France; 3 Univ Paris-Sud, 

Villejuif, France 

Background: Acute severe autoimmune hepatitis (AS-AIH) is 

a rare, heterogeneous disease, with a high rate of liver transplantation 

(LT) or death and a poor determination of prognostic 

factors. Aim: The aim of the present study was to describe the 

prognostic role of chronic hepatitis for the outcome of patients 

presenting with AS-AIH. Methods: Patients admitted to our center, 

between January 2009 and June 2014, with the diagnosis 

of AS-AIH were retrospectively identified. The AS-AIH diagnosis 

was based on: 1. IAIHG score definite or probable, 2. 

INR > 1.5, 3. No previous medical history of AIH, 4. Histological 

features of AIH. A single expert pathologist reviewed the 

liver biopsies. Patients with chronic hepatitis (CH) defined by 

a fibrosis stage 2, according to METAVIR, were compared to 

patients without chronic hepatitis (non-CH). Results: Fourteen 

patients were included, 11 (78.5%) female, median age 52 

[18-69] years. CH was diagnosed in 8/14 (57%) patients on 

liver biopsy, among them 4 were cirrhotic. The histological 

evaluation was performed on the explant for 3 patients, all of 

them without CH. Median age of CH patients was 57 [18-69] 

years versus (vs) 44 [18-66] of non-CH patients (p=ns). CH 

patients compared to non-CH patients had: lower MELD score, 

27 [16-32] vs 33 [24-34] (p=0.02), lower bilirubin, 237 

mmol/L [26-363] vs 378 [241-541] (p=0.03), lower median 

INR 2.77 [1.68-4.04] vs 3.75 [1.9-4.29] (p=0.12) although 

the difference was not statistically significant. Patients of both 

groups had similar IgG levels, 32.3 [22.2-61.7] vs 31.9 [9.67] 

(p=ns), and all but one had positive antinuclear or anti-smooth 

muscle antibodies. Eleven (78.5%) patients received steroids 

therapy (0.5 to 1 mg/kg/day): 7/8 patients with CH versus 

4/6 patients without. The median interval time between hospital 

admission and steroids therapy was 3 [0-119] days for CH 

patients vs 9 [2-69] days for non-CH pts (p=ns). Infections were 

detected in 37.5% (3/8) of CH patients vs 50% (3/6) non-CH 

pts (p=ns). All patients who did not receive steroids therapy 

underwent LT. LT was performed in 25% (n=2) of CH patients 

vs 83% (n=5) of non-CH pts (p=0.03). Overall survival was 

100%, with a median follow-up of 24 [3-69] months. Conclusions: 

In patients with severe acute autoimmune hepatitis histological 

assessment has a key prognostic role as the absence 

of chronic hepatitis predicts the need of liver transplantation, 

despite steroids therapy. 

Disclosures: 

Audrey Coilly - Speaking and Teaching: Gilead, BMS, Janssen, MSD, Roche, 

Novartis, Astellas 



The following authors have nothing to disclose: Eleonora De Martin, Mylene 

Sebagh, Badr Serji, Teresa Maria Antonini, Eric Ballot, Florent Artru, Philippe 

Ichai, Jean-Charles Duclos-Vallee 

1783 

Hemophagocytic lymphohistiocytosis (HLH) presenting 

as acute liver failure: three cases at a liver transplant 

center 

Amanda Schneier 1 , Ponni V. Perumalswami 2 , Sunyoung Lee 3 ; 

1 Medicine, Icahn School of Medicine at Mount Sinai, New York, 

NY; 2 Liver Diseases, Icahn School of Medicine at Mount Sinai, 

New York, NY; 3 Medicine, Elmhurst Hospital, Queens, NY 

OBJECTIVE: Hemophagocytic lymphohistiocytosis (HLH) is 

primarily known as a pediatric disease but has been rarely 

described in adults, usually in the setting of EBV infection, 

malignancy, rheumatologic disease, or as a side effect of 

immune-modulating medications. The pathophysiology of HLH 

has been described as an uncontrolled activation of the immune 

system and a cytokine storm, resulting in excessive inflammation 

and phagocytosis of patients’ own blood cells by histiocytes. 

Patients with HLH typically present with develop fever, 

organomegaly, lymphadenopathy, liver dysfunction, and cytopenias 

in the absence of infection. METHODS: This case series 

describes three patients at an urban, academic liver transplant 

center. All cases presented with acute liver failure and were ultimately 

diagnosed with HLH secondary to occult malignancies. 

The cases presented from February 2014 to February 2015. 

RESULTS: All three cases were female patients ranging from 44 

to 53 years old. All were transferred from other hospitals for 

workup of severe jaundice and liver dysfunction, suspected to 

be secondary to toxin-induced liver injury. The patients went on 

to develop severe fevers and refractory cytopenias. All three 

underwent a rapid evaluation for liver transplant and were evaluated 

by a multidisciplinary team including hepatology, critical 

care, infectious disease, and hematology. A complete work up 

for acute liver failure including viral serologies, autoimmune 

markers, and ceruloplasmin were negative for all three cases, 

and none had significant alcohol consumption or other drug or 

toxin exposure. The diagnosis of HLH was made after a mean 

of 10 days by bone marrow biopsy. Bone marrow pathology 

featured histiocytes with hemophagocytosis in all patients. Two 

out of three cases also had evidence of hemophagocytosis on 

liver pathology (one upon autopsy). The patients’ initial ferritin 

levels ranged from 4,170 to 48,000 ng/mL, and all eventually 

rose to >50,000 ng/mL. All cases were treated with HLH-directed 

therapy (two with etoposide, one with pentostatin) but 

expired during their hospitalization. All cases were found to 

have prior exposure to EBV but undetectable viral loads during 

admission, and were subsequently found to have occult malignancies: 

two with hepatosplenic T-cell lymphoma and one with 

papillary thyroid carcinoma. CONCLUSIONS: HLH can present 

as acute liver failure in adult patients. Given the low success 

rate of treatment, early diagnosis of HLH is critical. Therefore, 

a higher degree of suspicion should be exercised in otherwise 

unexplained rapidly progressing acute liver failure in patients. 

Disclosures: 

The following authors have nothing to disclose: Amanda Schneier, Ponni V. 

Perumalswami, Sunyoung Lee 

1784 

Single Center Experience: Hypothermia in Acute Liver 

Failure Introduction 

Edson S. Franco 2 , Alexia Makris 1 , Manohar M. Lahoti 1 , Angel 

Alsina 2 ; 1 Information Systems and Decision Sciences, University 

of South Florida, Tampa, FL; 2 Surgery, Tampa General Hospital, 

Tampa, FL 

Introduction Acute liver failure is a rare condition with a dismal 

prognosis in the setting of hepatic encephalopathy (HE).


Hypothermia (HT) has been shown to prevent cerebral edema 

and intracranial hypertension associated HE. In this study, we 

report the outcomes of ALF patients treated with hypothermia 

with and without transplantation. Methods This is a single 

center retrospective study from 2002 to 2014. We reviewed 

all ALF patients treated with HT for HE with and without liver 

transplantation (Oltx). Demographic data was collected. Primary 

and secondary outcomes include survival and complications 

respectively. Survival was calculated using Kaplan-Meier 

method. Bootstrap analysis was used to perform an additional 

internal validation. Results 44 patients were treated for ALF 

during the study period. The cohort was stratified as follows: 

Group 1 (HT/Oltx), Group 2 (HT/No Oltx), Group 3 (Medical 

Management/Oltx). The indication for HT was grade III/IV 

HE and instituted at 32°-34°. Twenty patients (46.5%) were 

bridged to transplantation. Group 2 patients were evaluated 

for Oltx except for one and 3 were listed. Survival was poor 

with mean 20 days. One died while waiting and 2 were 

removed. Four patients (17%) improved. Overall survival was 

50% (mean 362 days). One year-survival was 75% in Group 

1 versus 58% for Group 3 when patients received transplant. 

At the 2.5 years post Oltx, 50% (6 out of 12) in Group and 

62.5% (5 out of 8) in Group 1 were alive. Mean survival was 

584 days (SD 106) for Group 1, 294 (SD 49) for Group 3, 20 

(SD 3.4) for Group 2. Bootstrapping showed a difference in 

survival between Group 1 and Group 3 (Wilcoxon, Log-Rank, 

Likelihood Ratio p-Value 6 months, 

n=75); and G3, past HCV infection (ie, positive anti-HCV and 

negative HCV RNA PCR, n=15). RESULTS. In G1without HCV 

infection, 20 of 20 (100%) urine specimens tested negative for 

HCV-Ags using our EIA assay. Similarly, in G3 with past or 

resolved HCV infection, all 15 of 15 (100%) urine specimens 

tested negative for HCV-Ags, confirming 100% specificity of 

our HCV-Ags EIA when used for testing urine samples. Patients 

in G2 had chronic HCV genotype 1-6 infections, serum HCV 

RNA levels ranged from 62,000 to 9,960,000 IU/mL quantified 

using PCR, and serum creatinine level ranging from 0.5 

to 1.7 mg/dL. In 75 of 75 of G2 patients, all urine specimens 

tested positive for HCV-Ags, indicating our HCV-Ags assay had 

a sensitivity of 100% for testing urine specimens. The mean 

coefficient of variation (CVs) was 9.8% and 6.4%, respectively, 

for intra-assay precision and inter-assay reproducibility. 

Using a group of urine specimens from patients with very low 

serum HCV RNA PCR ranging from 26 to 148 IU/mL, the 

lowest detection limit of our HCV-Ags EIA for urine specimens 

was equivalent to serum HCV RNA level of 26 IU/mL. Optical 

density measurements for HCV-Ags from the urine specimens 

with chronic HCV infection correlated significantly with the 

corresponding serum HCV RNA level quantified using PCR (y 

= 0.0702 x + 0.141, R 2 = 0.8212, p


with positive antibody tests who did not have RNA testing and 

(c) 38,061 additional cases if the unscreened population in 

VA care was screened. Results for each step of the Cascade of 

HCV Care for 2013 and 2014 appear in the Table. Conclusion 

VA has diagnosed the majority of patients with HCV and linked 

them to HCV care. While antiviral treatment and SVR rates 

have improved in 2014 compared to 2013, the largest gap in 

care remains the initiation of antiviral treatment. 

VA Cascade of HCV Care 

HCV infection who were referred to care attended their first 

appointment and this was similar among non-Hispanic blacks 

and whites. Our findings suggest birth cohort testing is likely 

to identify a substantial number of persons living with HCV 

infection, although barriers persist early in the care continuum, 

particularly for non-Hispanic blacks in whom HCV infection 

was most prevalent. Additional efforts are needed to ensure 

confirmatory testing and linkage to care for all persons with an 

anti-HCV+ test. 

Disclosures: 

The following authors have nothing to disclose: Claudia Vellozzi, Rajiv Patel, Ben 

Schoenbachler, Susan Hariri 

Disclosures: 

The following authors have nothing to disclose: Lisa I. Backus, Pamela S. Belperio, 

Timothy P. Loomis, Larry A. Mole 

1787 

Hepatitis C Birth-Cohort Testing and Linkage to Care, 

Selected U.S. Sites, 2012-2014 

Claudia Vellozzi 1 , Rajiv Patel 2 , Ben Schoenbachler 2 , Susan 

Hariri 1 ; 1 CDC, Atlanta, GA; 2 ORISE, Oak Ridge, TN 

Objective To describe hepatitis C virus (HCV) infection prevalence 

and examine the provision of HCV testing and care by 

race/ethnicity among persons born during 1945-1965 seen 

at selected U.S. clinic settings primarily serving under/uninsured 

persons. Methods Data were drawn from CDC’s Hepatitis 

Testing and Linkage to Care project conducted at 105 

settings in 10 states from 2012-2014 to assess implementation 

of one-time HCV testing recommendation for persons born 

during 1945-1965. We examined anti-HCV positivity overall 

and by age, race/ethnicity, sex, and health insurance. Among 

persons who were anti-HCV+, testing and care was evaluated 

using the following indicators: HCV-RNA testing, chronic infection 

as determined by HCV-RNA positivity, referral to medical 

care, and attending first medical appointment. We further 

examined differences in HCV testing and care among non-Hispanic 

blacks compared to non-Hispanic whites. Results Among 

24,966 persons tested, 11.6% (2,900) were anti-HCV+; 

positivity was 17.1% in males (2,073/12,827), 13.9% in 

non-Hispanic blacks (1,701/12,202), and 16.0% in persons 

with public health insurance (1,868/11,650). Among all anti- 

HCV+ persons, 73% (2,108) received an HCV RNA test, 71% 

(1,497) were currently infected, 82% (1,223) were referred 

to care, and 77% (938) attended a first medical appointment. 

Among anti-HCV+ persons, the proportion receiving an RNA 

test was significantly higher among non-Hispanic whites compared 

to non-Hispanic blacks (87.6% vs. 63.2%; p < 0.0001). 

Conversely, a significantly larger proportion of non-Hispanic 

blacks were chronically infected compared to non-Hispanic 

whites (76.9% vs. 61.2%; p< 0.0001). There was no difference 

between non-Hispanic whites vs non-Hispanic blacks 

in attending a first medical appointment (75.0% vs. 79.2%; 

p=0.1954). Conclusion We found high (>11%) anti-HCV positivity 

among a population drawn from settings primarily serving 

under/uninsured persons. Overall, nearly 30% of individuals 

in this population did not receive an HCV-RNA test, and testing 

was disproportionately low in non-Hispanic blacks compared 

to non-Hispanic whites. Nearly 80% of all persons with current 

1788 

Treatment With Statins Reduces Liver Cancer Risk In 

Patients With Chronic Hepatitis C 

Anders H. Nyberg 1 , Ekaterina Sadikova 1 , Jiaxiao Shi 1 , T. Craig 

Cheetham 1 , Kevin Chiang 1 , Zobair M. Younossi 2 , Lisa M. 

Nyberg 1 ; 1 Kaiser Permanente, San Diego, CA; 2 Department of 

Medicine, Inova Faifax Medical Campus, Fairfax, VA 

Background: Statin use has been associated with a decreased 

risk of developing hepatocellular carcinoma (HCC) in patients 

with cardiovascular risk factors (1-2). An analysis of Taiwanese 

registry data by Tsan et al showed a decreased risk of 

developing HCC in patients with hepatitis C who were taking 

statins. However, there is a paucity of data in patients with 

liver disease. The aim of this study was to evaluate the effect 

of statin use on rates of HCC in our population of hepatitis C 

patients at Kaiser Permanente Southern California (KPSC), a 

community based health care system. Methods: A sample of 

35,712 patients >18 years of age with a chronic HCV diagnosis 

by ICD -9 code or positive HCV RNA test between 2002 

and 2012 was identified in the KPSC research database. The 

KPSC National Cancer Institute Surveillance, Epidemiology, 

and End Results (KPSC-NCI SEER) affiliated cancer registry was 

used to identify new HCC cases between 2008 and 2012. 

Statin use was defined as having 2 or more filled prescriptions 

prior to the outcome or the end of the follow-up. Clinical 

and sociodemographic patient characteristics were measured 

prior and during the study period. Crude and adjusted odds 

ratios of liver cancer associated with statin use were assessed 

using logistic regression modeling. Results: Statin use and liver 

cancer were independently associated with older age, race, 

higher prevalence of diabetes, liver cirrhosis, smoking, BMI 

and more total healthcare utilization. Crude and adjusted liver 

cancer logistic regression models are summarized below. Statin 

use prevailed as a strong protective effect after adjustment 

of clinical and sociodemographic characteristics. 

Disclosures: 




Lisa M. Nyberg - Grant/Research Support: Merck, Gilead, Abbvie


The following authors have nothing to disclose: Anders H. Nyberg, Ekaterina 

Sadikova, Jiaxiao Shi, Kevin Chiang 

1789 

Cause of death in HCV patients who achieved sustained 

virologic response: Chronic Hepatitis Cohort Study 

(CHeCS), 2006-2012 

Fujie Xu 1 , Jian Xing 1 , Anne C. Moorman 1 , Loralee B. Rupp 2 , Stuart 

C. Gordon 2 , Mei Lu 2 , Eyasu H. Teshale 1 , Philip R. Spradling 1 , 

Joseph A. Boscarino 3 , Connie M. Trinacty 4 , Mark A. Schmidt 5 , 

Scott D. Holmberg 1 ; 1 Division of Viral Hepatitis, CDC, Atlanta, 

GA; 2 Henry Ford Health System, Detroit, MI; 3 Geisinger Health 

System, Danville, PA; 4 Kaiser Permanente Hawaii, Honolulu, HI; 

5 Kaiser Permanente Northwest, Portland, OR 

Background: Successful treatment of chronic HCV infection is 

associated with reduced morbidity and mortality. The objective 

of this study was to describe the stage of liver disease at the 

time of sustained virologic response (SVR) and the timing and 

cause of death in post-SVR patients. Methods: We analyzed 

data from the Chronic Hepatitis Cohort Study (CHeCS), an 

ongoing ‘dynamic’ observational study of patients with chronic 

HCV conducted at four integrated health care systems located 

in the United States (Henry Ford Health System, Geisinger 

Clinic, Kaiser Permanente-Northwest, and Kaiser Permanente-Hawaii). 

The stage of liver disease was approximated by 

FIB-4 score within 3 months of the SVR date (at 12 weeks post 

treatment). Deaths that occurred in 2006-2012 were ascertained 

by matching cohort patient records to the most recent 

National Death Index, Social Security Death Index, or electronic 

state death registries. We examined the causes of death 

listed on death certificates in patients whose HCV treatment 

had resulted in SVR. Results: Of 15,158 HCV patients enrolled 

in CHeCS, HCV treatment resulted in SVR in 1,492 (9.8%) 

patients between 2000 and 2012. Of these 1,492 patients, 

785 (52.6%) had one or more FIB-4 scores measured within 

3 months of the SVR date: the median FIB-4 score was 1.75, 

and 24.8% had a FIB-4 score >3.25. During 2006-2012, 

2,314 (15.3%) deaths occurred in HCV cohort patients, and 

of these, 71 (3.1%) were in patients known to have achieved 

SVR (4.8% (71/1,492) of SVR patients died versus 16.4% 

(2243/13,666) of non-SVR patients, p3.25 prior to SVR; the mean time 

from SVR date to death was 4.9 years (95% CI, 3.5-6.2), 

ranging from 1.4 years to 9.3 years. Conclusion: In a large 

HCV cohort in the United States, only 10% had achieved SVR 

by the end of 2012. While mortality among patients who had 

achieved SVR was lower, premature death and liver cancer 

can still occur and may be related to the late stage of liver 

disease at the time of treatment and SVR. 

Disclosures: 




The following authors have nothing to disclose: Fujie Xu, Jian Xing, Anne C. 

Moorman, Loralee B. Rupp, Mei Lu, Eyasu H. Teshale, Philip R. Spradling, Joseph 

A. Boscarino, Connie M. Trinacty, Mark A. Schmidt, Scott D. Holmberg 

1790 

Development of End Stage Liver Disease, Hepatocellular 

Carcinoma and Liver-Related Death According to 

Biopsy-Confirmed Ishak Fibrosis Stage in the Hepatitis C 

Alaska Cohort Study (AK-HepC) 

Dana J. Bruden 1 , Lisa J. Townshend-Bulson 2 , James E. Gove 2 , Julia 

N. Plotnik 2 , Chriss E. Homan 2 , Annette Hewitt 2 , Michael Bruce 1 , 

Prabhu P. Gounder 1 , Youssef Barbour 2 , Brenna Simons-Petrusa 2 , 

Brian J. McMahon 2,1 ; 1 Arctic Investigations Program, Division of 

Preparedness and Emerging Infections, Centers for Disease Control 

and Prevention, Anchorage, AK; 2 Liver Disease and Hepatitis 

Program, Alaska Native Tribal Health Consortium, Anchorage, AK 

Background Most studies on liver biopsy results among hepatitis 

C virus (HCV) infected persons are of a cross-sectional study 

design. Long-term prospective studies of the outcomes associated 

with HCV are rare and critical for assessing the potential 

impact of HCV treatment on sequelae. We used liver biopsy 

as a study start point and looked at development of end stage 

liver disease (ESLD), hepatocellular carcinoma (HCC) and liver-related 

death (LRD) according to the Ishak fibrosis score. 

Methods Since 1994, all American Indian/Alaska Native (AI/ 

AN) persons positive for anti-HCV have been invited to enroll in 

a long-term study of HCV outcomes. By 2012, 1,325 persons 

were enrolled, of whom 1,080 were HCV RNA-positive, and 

of these, 412 had a liver biopsy performed. In persons with 

multiple liver biopsies, the most recent was used. The Ishak 

fibrosis score was used to categorize persons into mild/moderate 

(Ishak = 0, 1, 2), severe (Ishak = 3, 4) or cirrhosis (Ishak 

= 5, 6). The date of liver biopsy was the starting point in a 

survival analysis examining time until development of ESLD, 

HCC and LRD. We report survival probabilities at 5-years using 

Kaplan-Meier estimates. Results Of 412 persons undergoing 

liver biopsy, 68% (n = 282) had mild/moderate fibrosis, 21% 

(n = 87) had severe fibrosis and 10% (n = 43) had cirrhosis. 

The average length of following liver biopsy was 7.7 years 

with a total of 3,162 years of follow-up. The average age 

at the time of biopsy was 44.3 years and 51% (n = 211) 

were female. Within 5 years of biopsy, 4.6% (95% confidence 

interval (CI): 2.5, 8.5) of persons with mild/moderate fibrosis 

had developed ESLD compared to 16.4% (CI: 9.6, 27.2) and 

45.1% (CI: 29.7, 63.9) of persons with severe fibrosis and 

cirrhosis, respectively (p


1791 

Ability of EMR-based fibrosis scores to identify HCV-infected 

patients with cirrhosis 

Mohammad Qasim Khan 3 , Vijay Anand 3 , Ammar Hassan 3 , Alya 

Assan 3 , Amnon Sonnenberg 2 , Claus J. Fimmel 1 ; 1 Gastroenterology, 

Northshore University Health System, Evanston, IL; 2 Gastroenterology, 

Oregon Health Sciences University, Portland, OR; 

3 Internal Medicine, NorthShore University Health System, Evanston, 

IL 

Background: With the advent of fixed-dose treatment regimens 

for chronic HCV (CHC) infection and the updated, population-based 

screening guidelines, clinicians are faced with the 

challenge of assessing the degree of liver fibrosis in large 

patient populations. We asked whether it would be possible 

to predict cirrhosis in HCV-infected patients using fibrosis algorithms 

that are entirely based on simple laboratory and demographic 

features extractable from the electronic medical record 

system (EMR). Objective: To compare a set of seven EMR-derived 

fibrosis scores with liver imaging studies in a cohort of 

CHC patients. Methods: Patients with CHC (n=869) were identified 

by searching the EPIC-derived patient data warehouse. A 

total of 566 patients had undergone a liver imaging study, and 

had no confounding medical conditions affecting the fibrosis 

scores. Cirrhosis was defined as the presence of a nodular 

liver or portal collaterals on ultrasound, CT, or MRI imaging. 

Demographic and laboratory data were extracted from the 

EMR, matched with the date of the imaging studies, and used 

to calculate the following previously established fibrosis scores: 

APRI, Fib4, Fibrosis Index, Forns, GUCI, Lok Index, and Vira- 

HepC. Areas under the receiver operating curves (AUROC), 

optimum cut-offs, positive and negative predictive values were 

calculated for each score. Results: The seven algorithms performed 

similarly well in predicting cirrhosis (Table). No single 

test was superior as all of their AUROC confidence intervals 

overlapped. Sensitivites ranged from 0.65 to 1.00, specificities 

from 0.67 to 0.90, positive predictive values from 0.33 

to 0.38, and negative predictive values from 0.93 to 1.00. 

Summary: EMR-based algorithms performed well in ruling out 

radiologic cirrhosis, whereas their ability to predict the presence 

of cirrhosis was modest. Conclusions: Simple, EMR-based 

scoring systems can be used to rule out advanced cirrhosis in 

CHC patients with high reliability. They are less accurate in 

identifying patients with advanced cirrhosis, suggesting that 

positive scores would need to be confirmed by secondary testing. 

Table 

1792 

Characteristics of Patients Tested for Hepatitis C and 

Intervention Costs in the BEST-C Study 

Joanne E. Brady 1 , Danielle Liffmann 1 , Anthony K. Yartel 2 , Natalie 

B. Kil 3 , Alex D. Federman 3 , Cynthia E. Jordan 4 , Omar I. Massoud 4 , 

David R. Nerenz 5 , Kimberly Ann Brown 5 , Bryce Smith 6 , Claudia 

Vellozzi 2 , David B. Rein 1 ; 1 Public Health, NORC at the University 

of Chicago, Bethesda, MD; 2 Division of Viral Hepatitis, U.S. Centers 

for Disease Control and Prevention, Atlanta, GA; 3 Division 

of General Internal Medicine, Mount Sinai School of Medicine, 

New York, NY; 4 Department of Medicine, University of Alabama 

at Birmingham, Birmingham, AL; 5 Department of Medicine, Henry 

Ford Hospital, Detroit, MI; 6 Division of Diabetes Translation, U.S. 

Centers for Disease Control and Prevention, Atlanta, GA 

Given that 80% of Hepatitis C virus (HCV)-infected Americans 

were born during the years 1945-1965, the Centers for Disease 

Control and Prevention (CDC) and the U.S Preventive 

Services Task Force recommended a one-time HCV antibody 

test for adults born in the 1945-1965 birth cohort (BC). CDC’s 

Birth-cohort Evaluation to Advance Screening and Testing for 

Hepatitis C was designed to assess the impact of testing interventions 

on the probability of HCV testing in primary care 

(PC) among BC patients as compared to usual care and the 

incremental costs per person tested and per case identified 

at each site. From December 2012-March 2014, 3 health 

systems implemented independent testing interventions using 

randomized designs to compare intervention testing and identification 

rates to usual care. Site 1 mailed paid lab test orders 

and repeated reminders to a randomly selected list of active 

patients compared to a second list who received no mailings. 

Site 2 created an electronic health record best practice alert 

(BPA) implemented or not implemented based on cluster randomized 

design. Site 3 directly solicited patients following a 

scheduled PC visit and used a cluster randomized crossover 

design. Multilevel multivariable regression was used to estimate 

the risk ratio for HCV testing; activity-based costing was used 

to estimate costs. HCV testing was significantly more common 

for all interventions compared to controls; adjusted risk ratio 

(aRR) 19.2, (95% CI, 9.7–38.2), 13.2 (95% CI, 3.6–48.6), 

and 32.9 (95% CI 19.3–56.1) for sites 1, 2, and 3, respectively. 

The BPA intervention had the lowest incremental cost 

per person tested ($25 with fixed startup costs, $3 without 

startup costs). The incremental cost per new case identified 

under usual care ranged from $3,771-$6207 across sites. All 

interventions increased HCV testing among the BC compared 

to usual care, but also increased the costs. The cost per case 

identified excluding startup costs was lowest for the BPA intervention 

($1,691), suggesting that integrating BC testing into 

usual care is likely to be more cost-effective than instituting an 

intervention in addition to usual care, e.g., repeated-mailings 

and patient-solicitation. 

n=number of patients, AUROC=area under the receiver operating 

curve, CI=confidence interval 

Disclosures: 

The following authors have nothing to disclose: Mohammad Qasim Khan, Vijay 

Anand, Ammar Hassan, Alya Assan, Amnon Sonnenberg, Claus J. Fimmel 

Costs measured in dollars


Disclosures: 

Kimberly Ann Brown - Advisory Committees or Review Panels: CLDF, Merck, 

BMS, ABBVIE, Gilead, Gilead, Janssen, Salix; Consulting: Blue Cross Transplant 

Centers; Grant/Research Support: CLDF, Gilead, Exalenz, CDC, BMS, Hyperion, 

Merck; Speaking and Teaching: ABBVIE, Gilead, CLDF 

David B. Rein - Grant/Research Support: Gilead Sciences, Inc. 

The following authors have nothing to disclose: Joanne E. Brady, Danielle Liffmann, 

Anthony K. Yartel, Natalie B. Kil, Alex D. Federman, Cynthia E. Jordan, 

Omar I. Massoud, David R. Nerenz, Bryce Smith, Claudia Vellozzi 

1793 

Risk of hepatitis C virus infection in people who use 

drugs who report male-to-male sex 

Chloe Le Marchand 1 , Ali Mirzazadeh 1 , Stephen Shiboski 1 , Thomas 

M. Rice 1 , Meghan D. Morris 1 , Judith A. Hahn 1 , Kimberly Page 2,1 ; 

1 University of California, San Francisco, Berkeley, CA; 2 University 

of New Mexico, Albuquerque, NM 

Introduction Most new hepatitis C virus (HCV) infections occur 

among people who use drugs (PWID) via shared injecting 

equipment. Rising HCV infection rates in non-injecting, HIV-infected 

men who have sex with men (MSM) are linked to highrisk 

sexual activity. Studies examining the incidence of HCV in 

MSM have mixed results; several show an increased susceptibility 

to HCV principally in HIV-infected patients. Incidence 

rates range from 18.0/1000 person-years (py) in HIV-infected 

MSM 9.0/1000 py in MSM without HIV. In this study, we 

estimate HCV incidence in male PWID in association with 

MSM behavior, controlling for injecting exposures. Methods 

We used data from the INC3 collaboration, which pooled biological 

and behavioral data from 9 cohorts of PWID in North 

America, Europe and Australia, including 1,921 males at risk 

for HCV. We included all participants in INC3 with a history of 

drug use and a negative anti-HCV test at enrollment. We conducted 

contingency table analyses with Chi-square analysis for 

descriptive variables by MSM status. Incident infections were 

defined as a positive infection (anti-HCV positive or HCV RNA 

positive) occurring during follow-up. We used the log-rank test 

to compare survival curves by sexual and injecting risk behaviors. 

We estimated incidence by MSM status as compared to 

non-MSM males and conducted Cox regression analyses to 

compute hazard ratios controlling for potential confounders, 

including: HIV status, recent injecting, syringe and cooker sharing, 

injection frequency, number of injecting partners, access 

to clean needles, year entered the study and clustered by site. 

Results In 1,064 male participants, followed for 1,921 py, 

342 new HCV infections were observed for an overall incidence 

of 16.1/100 py (95% CI 14.4, 18.0). No differences 

were seen in estimated HCV incidence in MSM participants 

(16.3/100 py (95% CI 12.5, 21.2)) compared to non-MSM 

males (16.0/100 py (95% CI 14.1, 18.1)). We assessed HCV 

incidence in HIV-infected compared to non-infected groups and 

found no differences. MSM were different from non-MSM at 

baseline with respect to injecting risk: 61% vs. 47% reported 

syringe or equipment sharing, respectively (p


1795 

Distribution of pre-existing NS5A/NS5B resistance associated 

variants in genotype 1b patients with hepatitis C 

virus and response to direct acting antivirals 

Takeshi Watabe, Masaaki Korenaga, Masaya Sugiyama, Erina 

Kumagai, Misuzu Ueyama, Yoshihiko Aoki, Yoko Yamagiwa, 

Keiko Korenaga, Masatoshi Imamura, Kazumoto Murata, Naohiko 

Masaki, Tatsuya Kanto, Masashi Mizokami; The research center 

for hepatitis and Immunology, National Center of Global Health 

and Medicine (NCGM) at Kohnodai, Ichikawa, Japan 

Background and Aim: Although interferon-free antiviral treatment 

is improved sustained virological response (SVR) rate of 

hepatitis C virus (HCV), pre-existing NS5A/NS5B resistance 

associated variants (RAVs) may affect the efficiency of some 

HCV regimens. The aim of study is to investigate whether the 

NS5A/NS5B RAVs interact with clinical characteristics and 

response to 12 weeks of treatment with the nucleotide polymerase 

inhibitor sofosbuvir (SOF) combined with the NS5A 

inhibitor ledipasvir (LDV) for genotype (GT)-1b infection. Methods: 

Three hundred sixteen Japanese patients with HCV GT-1b 

(mean Age [ys]:67, male [%]:32, treatment naïve [%]:53 cirrhosis 

[%]:30, and hepatocellular carcinoma (HCC) [%]:10) 

were examined NS5A/NS5B RAVs by direct sequence. NS5A 

RAVs included L31M/I/V/F and Y93H, and NS5B RAVs 

included S282T and C316N. The frequency of the RAVs was 

examined the association with clinical characteristics, such as 

age, sex, naïve/experienced, HCVRNA, fibrosis, and hepatocellular 

carcinoma (HCC). Fibrosis was evaluated by Fibroscan 

and Mac-2 binding protein glycogen isomer. Among 

these patients, 59 patients (23%) who received LDV (90 mg)/ 

SOF (400 mg) for 12 weeks were analyzed the association 

of pre-treatment NS5A/NS5B RAVs with treatment outcome. 

Results: Distribution of NS5A RAVs existed 5% for L31 and 

23% for Y93. Double mutations in NS5A were detected only 

5 patients (1.5%). Distribution of NS5B RAVs existed 48% for 

C316; however there were no detection for S282 mutation. No 

association of these RAVs distribution with clinical characteristics. 

Fifty nine patients with receiving SOF/LDV were detected 

23% for NS5A RAVs, 48% for C316N, and 11% for NS5A 

RAVs with C316N and All 59 patients achieved SVR (100%). 

Conclusions: Although pre-treatment NS5A and NS5B RAVs in 

Japanese patients with infected HCV GT-1b exist about 25% 

and 50% respectively, presence of single RAVs, and NS5A 

RAVs with C316N in NS5B are less association with clinical 

characteristics, and treatment outcome of SOF/LDV. 

# All 59 patinets trerated with SOF/LDV for 12 weeks and 

achieved SVR 

Disclosures: 

The following authors have nothing to disclose: Takeshi Watabe, Masaaki 

Korenaga, Masaya Sugiyama, Erina Kumagai, Misuzu Ueyama, Yoshihiko Aoki, 

Yoko Yamagiwa, Keiko Korenaga, Masatoshi Imamura, Kazumoto Murata, Naohiko 

Masaki, Tatsuya Kanto, Masashi Mizokami 

1796 

Prevalence of viral hepatitis and liver fibrosis in streetbased 

outreach: a French prospective multicentre study 

Juliette Foucher 13 , Jean Harbonnier 1 , Leon Gomberoff 2 , Sylvie 

Balteau 3 , Fadi Meroueh 4 , Beatrice Stambul 5 , Rolland Le Hello 6 , 

Brigitte Reiller 7 , Xavier Richen 8 , Elisabeth Avril 9 , Jean-Pierre 

Daulouede 10 , Anne Borgne 11 , Veronique Latour 12 , Jean-Baptiste 

Hiriart 13 , Victor de Ledinghen 13 ; 1 CSAPA Boris Vian, Lille, France; 

2 CSAPA Ego Aurore, Paris, France; 3 CSAPA Les Wads, Metz, 

France; 4 CSAPA Arc en Ciel, Montpellier, France; 5 CSAPA Villa 

Floreal, Aix en Provence, France; 6 CSAPA L’envol, Rennes, France; 

7 CEID CSAPA Planterose, Bordeaux, France; 8 CSAPA du griffon 

Aria, Lyon, France; 9 CSAPA Gaia, Paris, France; 10 CSAPA Bizia 

Medecin du Monde, Bayonne, France; 11 service d’addictologie 

Hopital Muret, Sevran, France; 12 CSAPA La case Medecins du 

monde, Bordeaux, France; 13 hepatology unit, Bordeaux, France 

Introduction French Association for the Study of the Liver diseases 

guidelines recommends treatment of drug users whatever 

liver fibrosis stage. The aim of this prospective multicentre 

study was to evaluate viral hepatitis in French street-based outreaches 

and their severity. Patients and methods From January 

2012 to March 2013, all consecutive subjects with intravenous 

drug use (PWID) without any HCV management and consulting 

in street-based outreach in France were included. After face-toface 

questionnaire by outreach worker, liver stiffness was measured 

by FibroScan (FS). At the end of FS, a blood sample for 

hepatitis B and C serology was proposed as a meeting with a 

practitioner. During face-to-face interview, socio-demographic 

parameters and way of living were recorded. During the medical 

meeting, the result of serology and FS were explained. If 

case of HCV infection, users could meet a hepatologist and 

treatment could be proposed. Results In 12 French street-based 

outreaches, 860 drug-users were included (663 men, mean 

age 37 years, BMI 23 kg/m 2 , injected drug used history 

n=447, currently opioid substitution 62%, homeless 16,%, 

unemployed 72%, past history of incarceration 43%). 813 

subject accepted FS, with correct measurement in 94.5% of 

cases. Median liver stiffness was 5.1 kPa (2-75 kPa). 16.4% of 

users had significant fibrosis (FS>7.6 kPa) and 4.8% cirrhosis 

(FS>13 kPa). 77% of drug users with interpretable FS had a 

blood sample. HCV prevalence was 34%. HIV and HBV prevalence 

were 2% and 1.2%, respectively. Factors associated with 

HCV infection were age, history of IVDU, history of incarceration 

and no insurance. In multivariate analysis, factors associated 

with HCV were history of IVUD (OR 19.38, [9.9-37.8] 

p


The following authors have nothing to disclose: Jean Harbonnier, Leon Gomberoff, 

Sylvie Balteau, Fadi Meroueh, Beatrice Stambul, Rolland Le Hello, Brigitte 

Reiller, Xavier Richen, Elisabeth Avril, Jean-Pierre Daulouede, Anne Borgne, 

Veronique Latour, Jean-Baptiste Hiriart 

1797 

Hepatitis C RNA assay differences in results around 6 

million IU/mL: Potential clinical implications for shortened 

Ledipasvir/Sofosbuvir therapy 

Gavin A. Cloherty 1 , Christoph Sarrazin 2 , Vera Holzmayer 3 , Jordan 

J. Feld 4 , Benjamin Maasoumy 5 , Johannes Vermehren 2 , Stephane 

Chevaliez 6 , Heiner Wedemeyer 5 , Jean-Michel Pawlotsky 6 , George 

Dawson 3 ; 1 R&D, Abbott Molecular, Des Plaines, IL; 2 Goeth Universtiy, 

Frankfurt, Germany; 3 R&D, Abbott Diagnostics, Abbott Park, 

IL; 4 Universtiy of Toronto, Toronto, ON, Canada; 5 Hannover Medical 

School, Hannover, Germany; 6 Hopital Henri Mondor, Paris, 

France 

Background/Aims:FDA approval of Ledipasvir/Sofosbuvir for 

the treatment of hepatitis C (HCV) includes the truncation of 

therapy from 12 to 8 weeks in treatment naïve, non-cirrhotic 

patients with HCV RNA levels 6 million and ART 3 on liver biopsy 

or APRI >1.5), ELSD (presence of ascites, esophageal varices, 

hepatic encephalopathy, or coagulopathy), HCC, and LRD 

(including liver transplantation) by using a Cox proportional 

hazards model. The model for advanced fibrosis and ESLD was 

adjusted for HCV genotype (1, 2, and 3), age at cohort entry, 

heavy alcohol use (average >50grams/day), obesity (body 

mass index [BMI] >30), and type II diabetes (DM2). The model 

for HCC risk was adjusted for HCV genotype, age at cohort 

entry, and sex. The model for LRD risk was adjusted for HCV 

genotype, age at cohort entry, and heavy alcohol use. Of the 

1068 participants, 66%, 19%, and 14% were infected with 

HCV genotype 1, 2, and 3, respectively. Compared with genotype 

1, genotype 3 was significantly associated with advanced 

fibrosis, ESLD, HCC and LRD (Table). Other age-adjusted significant 

risk factors for developing advanced fibrosis were heavy 

alcohol use (aHR: 2.0; CI: 1.5–2.7), obesity (aHR: 1.3; CI: 

1.0–1.7; p = 0.04), and DM2 (aHR: 1.6; CI: 1.1–2.2). Significant 

risk factors for developing ESLD were heavy alcohol use 

(aHR: 2.2; CI: 1.6–3.2), obesity (aHR: 1.4; CI: 1.0–1.9; p = 

0.03), and DM2 (aHR: 1.5; CI: 1.0–2.3; p = 0.03). Another 

significant risk factor for developing HCC was male sex (aHR: 

3.6; CI: 1.6–8.2) and for developing LRD was heavy alcohol 

use (aHR: 2.9; CI: 1.8–4.6). At 15 years of follow-up, twothirds 

(68%) with genotype 3 were predicted to have developed 

advanced fibrosis and half (48%) ESLD. Conclusions:


Because AI/AN persons infected with HCV genotype 3 are at 

high risk for developing advanced fibrosis, ESLD, HCC, and 

LRD, they should be prioritized for earlier antiviral treatment. 

Disclosures: 

The following authors have nothing to disclose: Anne C. Moorman, Yueren Zhou, 

Loralee B. Rupp, Joseph A. Boscarino, Connie M. Trinacty, Scott D. Holmberg 

Disclosures: 

Lisa J. Townshend-Bulson - Grant/Research Support: Gilead 

James E. Gove - Grant/Research Support: Gilead Sciences 

Annette Hewitt - Grant/Research Support: gilead 

Julia N. Plotnik - Grant/Research Support: Gilead 

Chriss E. Homan - Grant/Research Support: Gilead 

Hannah G. Espera - Grant/Research Support: Gilead 

Youssef Barbour - Grant/Research Support: Gilead 

The following authors have nothing to disclose: Brian J. McMahon, Dana J. 

Bruden, Stephen Livingston, Susan Negus, Mary Snowball, Michael Bruce, Philip 

R. Spradling, Prabhu P. Gounder 

1799 

The Utility and Limitations of Electronic Medical Records: 

Predictive Values of ICD9 Codes indicating Chronic HCV 

Infection 

Anne C. Moorman 1 , Yueren Zhou 2 , Loralee B. Rupp 2 , Joseph A. 

Boscarino 3 , Connie M. Trinacty 4 , Scott D. Holmberg 1 ; 1 Division of 

Viral Hepatitis, C DC, Atlanta, GA; 2 Henry Ford Health Systems, 

Detroit, MI; 3 Geaisinger Health System, Danville, PA; 4 Kaiser Permanente-Hawaii, 

Honolulu, HI 

Objective. With increasing analysis of electronic medical 

records (EMR) in the United States, the predictive values of 

the International Classification of Diseases, 9th revision (ICD- 

9) coding system for chronic hepatitis C virus (HCV) infection 

warrant evaluation. Methods. Patients in the Chronic Hepatitis 

Cohort Study (CCHeCS) with chronic HCV infection, confirmed 

by laboratory data and abstractor review (a ‘gold standard’), 

were compared to “chronically infected cases” identified by 

electronic medical records (EMRs) ICD9 codes of adult patients 

who accessed services from 2006 to 2012 at four integrated 

healthcare systems in the United States. Results. Of over 1.6 

million adult patients at the 4 large medical systems in CHeCS, 

11,354 were confirmed as having chronic HCV infection. 

Having an EMR with one ICD-9 code for HCV yielded good 

sensitivity and positive predictive value (PPV) for true HCV 

infection, but poor specificity (Table), ie.e many (n=724) who 

were listed as having HCV infection but were not infected. 

Use of two hepatitis C-specific ICD-9 codes (6 or more months 

apart) improved the specificity with some loss of sensitivity, 

and the negative predictive value (NPV, non-cases accurately 

identified as non-cases) was poor (Table) . Because test codes 

were not standardized between laboratories, and viral load 

results were in text fields, distinguishing true from questionable 

or non-cases of HCV infection required extensive visual inspection 

and data processing. Discussion. EMRs, especially ICD9 

(or ICD10) codes, are increasingly used and analyzed in the 

United States, but these have imperfect sensitivity, specificity, 

and positive and negative predictive values for hepatitis C, and 

require considerable human review and judgment. 

1800 

Liver fibrosis stage and comorbidities in persons with 

diagnosed hepatitis C infection, Chronic Hepatitis Cohort 

Study (CHeCS) 

Fujie Xu 1 , Jian Xing 1 , Anne C. Moorman 1 , Stuart C. Gordon 2 , 

Loralee B. Rupp 2 , Mei Lu 2 , Philip R. Spradling 1 , Eyasu H. Teshale 1 , 

Joseph A. Boscarino 3 , Mark A. Schmidt 4 , Connie M. Trinacty 5 , 

Scott D. Holmberg 1 ; 1 Division of Viral Hepatitis, Centers for Disease 

Control and Prevention, Atlanta, GA; 2 Henry Ford Health System, 

Detroit, MI; 3 Geisinger Health System, Danville, PA; 4 Kaiser 

Permanente- Northwest, Portland, OR; 5 Kaiser Permanente-Hawaii, 

Honolulu, HI 

Background and Aims: The Chronic Hepatitis Cohort Study 

(CHeCS), a dynamic, prospective, observational cohort study, 

was created to study the natural history of chronic viral hepatitis 

in the United States. This report described HCV cohort patients’ 

demographic and clinical characteristics, comorbidities, treatment 

and treatment outcomes of cohort patients at the end of 

2012. Methods: CHeCS patients were drawn from about 2.7 

million persons aged 18 years or older who had a clinical service 

visit from January 2006-December 2012 at 4 integrated 

healthcare systems in Detroit, Michigan; Danville, Pennsylvania; 

Portland, Oregon; and Honolulu, Hawaii. We analyzed 

clinical data obtained from electronic medical records and 

manual chart review for HCV-infected patients who were alive 

but never had a liver transplant as of December 2012, including 

patients who achieved SVR with altered natural history. 

Patient’s current fibrosis stage was based on latest FIB4 scores 

derived from blood tests or biopsy results, if available. Laboratory 

tests and ICD-9 codes were used to identify patients with 

co-infections and comorbidities. Results: Of 15,940 total HCV 

cohort patients 2006-12, 11,294 were alive of Dec 2012; 

among these patients mean age at the end of observation was 

54 years; 58% were male, 55% white and 22% black. Only 

1,355 (9%) of the cohort had achieved SVR, which occurred 

a median of 4.8 years previously. Of 9,975 patients without 

SVR, 2,553 (25.6%) had ever been treated during a median 

followup of 5.3 years. Viral load data was available for 7,189 

patients; of these, 14% had 6 million IU/ml or higher (most 

recent test among those untreated and the latest before initial 

treatment among those treated). Impaired kidney function with 

eGFR


Disclosures: 




The following authors have nothing to disclose: Fujie Xu, Jian Xing, Anne C. 

Moorman, Loralee B. Rupp, Mei Lu, Philip R. Spradling, Eyasu H. Teshale, Joseph 

A. Boscarino, Mark A. Schmidt, Connie M. Trinacty, Scott D. Holmberg 

1801 

From Care to a Cure: Improving the Hepatitis C Care 

Cascade through Patient Navigation in the Check Hep C 

Program in New York City 

Mary Ford, Nirah Johnson, Payal Desai, Eric J. Rude, Fabienne 

Laraque; New York City Department of Health and Mental 

Hygiene, New York, NY 

Background: Of the estimated 146,500 persons with hepatitis 

C (HCV) infection in New York City (NYC), only about 10% are 

thought to have ever received treatment. New, highly effective 

HCV treatments provide an impetus for increased public health 

efforts to support HCV-infected persons along the care cascade 

from diagnosis to cure. Check Hep C, a program of the NYC 

Department of Health and Mental Hygiene (DOHMH), provided 

HCV screening, navigation, and tele-medicine in Year 1. 

Over 4,000 persons were tested, and 512 were RNA positive. 

Of those, 435 (85%) were linked to care, 14 initiated treatment 

and 3 had a sustained virological response (SVR). The small 

number of patients treated and cured highlighted the need 

for intensified patient navigation services through treatment. 

Methods: In Year 2, navigation services were provided at four 

sites following a standardized protocol. Services included a 

comprehensive social and health assessment, appointment 

assistance, support for medical evaluation and treatment, and 

pharmacy coordination. Patient navigators worked to enroll a 

target of 400 participants and form strong relationships with 

the multidisciplinary team involved in each patient’s care. Program 

data were entered in a standard database by the patient 

navigators and sent to DOHMH. Descriptive analysis was conducted. 

Results: Between March 2014 and January 2015, 388 

(97% of the target) participants were enrolled in Check Hep C. 

The median age of participants was 52 years, and 236 (61%) 

were born between 1945-1965. Of 376 participants with 

race/ethnicity data, 242 (64%) were Hispanic and 103 (27%) 

were Black, non-Hispanic. Of 364 participants with risk history 

data, 182 (50%) had current chemical dependence, 108 

(30%) had a history of intravenous drug use, and 91 (25%) 

were homeless. As of April 2015, 308 (79%) participants 

received HCV medical care, and 301 (78%) completed an 

HCV medical evaluation, of which 232 (77%) were treatment 

candidates. Of the treatment candidates, 116 (50%) initiated 

treatment, and 86 (74%) of those completed treatment. Thus 

far, 70 participants (81% of those who completed treatment) 

achieved SVR. Conclusion: The Check Hep C program successfully 

assisted high-need participants through HCV care and 

treatment, including those with active chemical dependence 

and homelessness, helping a much higher number of patients 

in achieving SVR in Year 2. Programs such as this community-based 

patient navigation intervention can help improve the 

HCV care cascade, particularly for high-need persons. Sustainable 

sources of funding, including insurance reimbursement, 

need to be provided to support these critical services. 

Disclosures: 

Eric J. Rude - Grant/Research Support: Vertex, Merck, Bristol Myers Squibb, 

Orasure, Janssen, Gilead, Kadmon, Boehringer-Ingelheim, Abbott, Genentech 

The following authors have nothing to disclose: Mary Ford, Nirah Johnson, Payal 

Desai, Fabienne Laraque 

1802 

Treatment prioritization according to the EASL HCV CPG 

2015: a real-life evaluation on the PITER (Piattaforma 

Italiana per lo studio della Terapia delle Epatiti viRali) 

cohort 

Loreta A. Kondili 1 , Stefano Rosato 2 , Maria Giovanna Quaranta 1 , 

Liliana E. Weimer 1 , Loredana Falzano 1 , Alessandra Mallano 1 , 

Maria Elena Tosti 2 , Maurizio Massella 1 , Maurizia R. Brunetto 3 , 

Anna Linda Zignego 4 , Mario Rizzetto 5 , Alfredo Di Leo 6 , Giovanni 

Raimondo 7 , Carlo Ferrari 8 , Gloria Taliani 9 , Pierluigi Blanc 28 , Antonio 

Gasbarrini 10 , Luchino Chessa 11 , Elke M. Erne 12 , Giovanna Fattovich 

13 , Pietro Andreone 14 , Maria Vinci 15 , Francesco P. Russo 16 , 

Erica Villa 17 , Giovanni B. Gaeta 18 , Teresa A. Santantonio 19 , Guglielmo 

Borgia 20 , Gabriella Verucchi 21 , Carmine Coppola 22 , Marcello 

Persico 23 , Liliana Chemello 29 , Alfredo Alberti 16 , Vincenzo De 

Maria 30 , Massimo Puoti 31 , Raffaele Bruno 24 , Paolo Caraceni 14 , 

Massimo Andreoni 25 , Marco Marzioni 26 , Stefano Vella 1 , Antonio 

Craxi 27 ; 1 Therapeutic Research and Medicines Evaluation, 

Istituto Superiore di Sanità, Rome, Italy; 2 Istituto Superiore di Sanità, 

Rome, Italy; 3 Azienda Ospedaliero-Universitaria Pisana, Pisa, 

Italy; 4 University of Florence, Florence, Italy; 5 University of Torino, 

Torino, Italy; 6 University of Bari, Bari, Italy; 7 University Hospital of 

Messina, Messina, Italy; 8 Azienda Ospedaliero- Universitaria di 

Parma, Parma, Italy; 9 Sapienza University of Rome, Rome, Italy; 

10 Policlinico Universitario Agostino Gemelli, Rome, Italy; 11 University 

of Cagliari, Cagliari, Italy; 12 Azienda Ospedaliera Padova, 

Padova, Italy; 13 University of Verona, Verona, Italy; 14 University 

of Bologna, Bologna, Italy; 15 Niguarda Hospital, Milan, Italy; 

16 Azienda Ospedaliero-Universitario Padova, Padova, Italy; 17 University 

of Modena and Reggio Emilia, Modena, Italy; 18 Second 

University of Naples, Naples, Italy; 19 University of Foggia, Foggia, 

Italy; 20 Federico II University of Naples, Naples, Italy; 21 Sant’Orsola 

Malpighi Hospital of Bologna, Bolgna, Italy; 22 Gragnano 

Hospital, Naples, Italy; 23 University of Salerno, Salerno, Italy; 

24 University of Pavia, Pavia, Italy; 25 Tor Vergata University, 

Roma, Italy; 26 Università Politecnica delle Marche, Ancona, Italy; 

27 University of Palermo, Palermo, Italy; 28 Santa Maria Annunziata 

Hospital, Florence, Italy; 29 University of Padua, Padua, Italy; 

30 Azienda Ospedaliero-Universitaria Mater Domini, Catanzaro, 

Italy; 31 Azienda Ospedaliera Niguarda-Cà Granda, Milano, Italy 

Aim. The high cost of DAAs for chronic hepatitis C has generated 

allocation policies mostly based on fibrosis staging as 

a surrogate for treatment needs. The EASL HCV CPG 2015 

indicates treatment allocation: treatment prioritized as first line, 

treatment justified as second line then deferred and not recommended 

treatment. We assessed the impact of this approach 

on a real-life cohort of 6831 consecutive patients presenting 

for care at 80 Italian Units, collected over the last 12 months 

within the PITER framework. Methods. The EASL CPG treatment 

prioritization algorithm considers fibrosis stage, HIV or HBV 

coinfection, pre and post transplant setting, extra-hepatic manifestations, 

i.v. drug use, hemodialysis. The independent effect 

of each of these factors and of major comorbidities (cardiovascular, 

metabolic, autoimmune/reumatological, neurological/psychiatric, 

neoplastic severe diseases) in patients older 

than 75 years, was evaluated by multivariate analysis. Results. 

Median age of the 6831 patients was 58 years (range 20-95 

yrs); 3797 (56%) were male. Matching of patients with the 

EASL CPG treatment indications resulted being categorized as 

follow: prioritized: 739 (11%) patients with F3 fibrosis; 1846 

(27%) patients with F4 fibrosis/Child A cirrhosis; 723 (11%) 

patients with Child B/C cirrhosis of whom 340 (5%) patients 

with HCC; 173 (3%) women of childbearing age; 280 (0.4%) 

drug users; 397 (6%) HIV or HBV coinfected patients; 587 

(9%) patients with severe extra-hepatic manifestations; 52 (1%) 

patients in LT waiting list; 84 (1%) patients with post LT HCV


recurrence. The following were categorized as justified: 440 

(6.4%) patients with F2 fibrosis; 661(10%) patients with F0-F1 

fibrosis fall in the deferrable category. Hence, in this large 

cohort, treatment should overall be prioritized in 3417 (50%) 

patients (HCV Gt 1: 66%; Gt 2: 10%; Gt 3: 12%; Gt 4: 7%), 

can be justified in 440 (6.4%) and deferred in 661 (9.6%) 

patients, while it is not indicated in 2313 (33.8%) patients due 

to severe co-morbidities or age older than 75 years. By comparison, 

the prioritization algorithm endorsed by AIFA (the Italian 

Medicines Agency), mostly based on fibrosis alone, which 

dictates reimbursement of DAA in Italy, would allow treatment 

of only 2827 (41%) of these patient cohort. Conclusion: Among 

patients with chronic hepatitis C, allocation of DAA according 

to priority rules linked to a perceived need for more prompt 

treatment will allow access to therapy to 40-55% of patients. 

There is a clear need for a validated predictive model to assess 

the impact of delaying treatment or not treating the remaining 

patients. 

Disclosures: 

Maurizia R. Brunetto - Speaking and Teaching: Roche, Gilead, Schering-Plough, 

Bristol-Myers Squibb, Abbott, Roche, Gilead, MSD, Novartis 

Mario Rizzetto - Advisory Committees or Review Panels: Merck, Janssen, BMS 

Alfredo Di Leo - Speaking and Teaching: Abbvie, MSD, Gilead, Roche, Italfarmaco, 

THD, CMD pharmalimited 



Carlo Ferrari - Advisory Committees or Review Panels: Gilead, Roche, Abbvie, 

BMS, Merck, Arrowhead; Grant/Research Support: Gilead, Roche, Janssen 

Gloria Taliani - Speaking and Teaching: ROCHE, Merck, BMS, Gilead, Jannsen, 

Novartis, AbbVie 


Jannsen 

Pietro Andreone - Advisory Committees or Review Panels: Janssen-Cilag, Gilead, 

MSD/Schering-Plough, Abbvie; Speaking and Teaching: Gilead, BMS 

Erica Villa - Advisory Committees or Review Panels: MSD, Abbvie, GSK, Gilead; 

Speaking and Teaching: Novartis 



Alfredo Alberti - Advisory Committees or Review Panels: Merck, roche, Gilead, 

Merck, roche, Gilead, Merck, roche, Gilead, Merck, roche, Gilead; Grant/ 

Research Support: Merck, gilead, Merck, gilead, Merck, gilead, Merck, gilead; 

Speaking and Teaching: novartis, BMS, novartis, BMS, novartis, BMS, novartis, 

BMS 

Massimo Puoti - Advisory Committees or Review Panels: GSK, Abbott, Janssen, 

MSD, Roche, Gilead Sciences, Novartis, GSK, Abbott, Janssen, MSD, Roche, 

Gilead Sciences, Novartis, GSK, Abbott, Janssen, MSD, Roche, Gilead Sciences, 

Novartis, GSK, Abbott, Janssen, MSD, Roche, Gilead Sciences, Novartis; Speaking 

and Teaching: BMS, BMS, BMS, BMS 



The following authors have nothing to disclose: Loreta A. Kondili, Stefano Rosato, 

Maria Giovanna Quaranta, Liliana E. Weimer, Loredana Falzano, Alessandra 

Mallano, Maria Elena Tosti, Maurizio Massella, Anna Linda Zignego, Pierluigi 

Blanc, Antonio Gasbarrini, Elke M. Erne, Giovanna Fattovich, Maria Vinci, Francesco 

P. Russo, Teresa A. Santantonio, Guglielmo Borgia, Gabriella Verucchi, 

Carmine Coppola, Marcello Persico, Liliana Chemello, Vincenzo De Maria, Raffaele 

Bruno, Massimo Andreoni, Marco Marzioni, Stefano Vella, Antonio Craxi 

1803 

Hepatitis E in haematological and rheumatological 

patients, a multicentre study 

Johann von Felden 1 , Laurent Alric 2 , Vincent Mallet 11 , Ansgar W. 

Lohse 1 , Paul Schnitzler 3 , Heiner Wedemeyer 4 , Christoph Hoener 

zu Siederdissen 4 , Maria Teresa Giordani 5 , Celia Aitken 6 , Jan 

Cornelissen 10 , Dominik Bettinger 7 , Robert Thimme 7 , Jean-Marie 

Peron 8 , Sven Pischke 1 , Robert A. de Man 9 ; 1 Gastroenterology, 

Hepatology, and Infectious Diseases, University Medical Centre 

Hamburg, Hamburg, Germany; 2 Unité de recherche clinique sur 

les hépatites virales, Médecine Interne-Pôle Digestif, CHU Purpan, 

Toulouse, France; 3 Virology, University Hospital, Heidelberg, Germany; 

4 Gastroenterology, Hepatology, Endocrinology, Hannover 

Medical School, Hannover, Germany; 5 Infectious and Tropical 

Diseases Unit, San Bartolo Hosptial, Vincenza, Italy; 6 Virology, 

NHS Greater Glasgow and Clyde, Glasgow, United Kingdom; 

7 Dept. Internal Medicine II, University Medical Centre Freiburg, 

Freiburg, Germany; 8 Gastro-entérologie et hépatologie, Médicine 

Interne-Pôle Digestif, Toulouse, France; 9 Hepatology, Gastroenterology, 

and Infectious Diseases, Erasmus Medical Center, 

Rotterdam, Netherlands; 10 Dept. Haematology, Erasmus Medical 

Center, Rotterdam, Netherlands; 11 Unité d’hépatologie, Université 

Paris Descartes, Paris, France 

Background Acute and chronic hepatitis E (HEV) in solid 

organ transplant recipients has been frequently described 

but the knowledge about HEV in patients with underlying 

haematological or rheumatological disease is limited. Methods 

We investigated retrospectively the clinical course of 47 

HEV infections in haematological (n=38) or rheumatological 

patients (n=9) within a multicentre observational study. Results 

Patients suffered from the following diseases: lymphoma n=17, 

myeloma n=3, chronic leukemia n=7, acute leukemia n=4, 

other haematological diseases n=7, rheumatoid arthritis, n=5, 

other rheumatological diseases=4. All patients received standard 

immunosuppressing therapy (calcineurin inhibitors: n=8, 

R-CHOP/R-Benda/R-ICE: n=7, cyclophosphamide n=7, other: 

n=28). 12/47 patients were stem cell transplant recipients 

(mean date of transplantation: 832 days before positive testing 

for HEV, range 20-4835 days). All patients tested positive for 

HEV RNA, and had laboratory signs of viral hepatitis (mean 

ALT 866 IU/ml, range 66-2849 IU/ml), while peak bilirubin 

was mean 7.3 mg/dl and peak creatinine was 90.2 mmol/l. 

All but one HEV infection had been acquired in Europe (1 

from Indonesia). Treatment with ribavirin (RBV) was initiated 

in 23/47 patients (5-22mg/kg bodyweight, duration: 1-32 

weeks, mean 14 weeks). Start of treatment ranged from immediately 

at the time of diagnosis up to 24 weeks after first detection 

of HEV (mean 9 weeks). 14/23 treated patients cleared 

the infection (61%), four patients (17%) failed to achieve viral 

clearance, one died (from deteriorating GvHd under treatment), 

two are still treated at the time of writing. 19 of the 24 

non-treated patients (79%) cleared the infection spontaneously, 

three died (two died directly after hepatitis E diagnosis from 

leukemia associated problems, one from GvHd), two are still 

under observation. RBV vs. untreated patients did not differ 

in peak ALT levels (RBV 836 IU/l, range 66-2775 IU/l vs. no 

treatment 931 IU/l, range 72-2849 IU/l), restitution of ALT 

after HEV infection (RBV 28 IU/l, range 11-85 IU/l (one patient 

with 937 IU/l) vs. no treatment 27 IU/l, range 13-76 IU/l) 

or in duration of HEV infection (RBV 27.1 weeks vs. no treatment 

27.5 weeks). ALT levels normalized in 37/47 patients 

(79%). Conclusion Chronic HEV infection is not restricted to 

solid organ transplant recipients but can also occur in immunosuppressed 

patients with haematological or rheumatological 

diseases. Ribavirin seems to be an efficient and safe treatment 

option. However, parameters that predict spontaneous clear-


ance need to be established in order to develop standards for 

initiating ribavirin therapy in immunosuppressed patients. 

Disclosures: 




MSD, Abbvie 





Jean-Marie Peron - Board Membership: BAYER; Consulting: BMS, GILEAD, BOS- 

TON SCIENTIFIC 



The following authors have nothing to disclose: Johann von Felden, Vincent 

Mallet, Ansgar W. Lohse, Paul Schnitzler, Christoph Hoener zu Siederdissen, 

Maria Teresa Giordani, Celia Aitken, Jan Cornelissen, Dominik Bettinger, Robert 

Thimme, Sven Pischke 

1804 

Linkage to Care Outcomes for Hepatitis C Cases Identified 

in the Emergency Department 

Ricardo A. Franco, Edgar T. Overton, Ashutosh Tamhane, Jordan 

M. Forsythe, Joel B. Rodgers, Deanne Guthrie, Suneetha Thogaripally, 

Anne Zinski, Michael Saag, Michael Mugavero, James W. 

Galbraith; University of Alabama at Birmingham, Birmingham, AL 

Purpose: Screening is an essential strategy for hepatitis C detection 

and urban Emergency Departments (EDs) are high-yield 

testing sites. While a critical step in the Care Cascade, no prior 

efforts longitudinally evaluated linkage to care (LTC) outcomes 

in ED screening programs. This study reports interim results of 

integrated LTC interventions in this setting. Methods: Consecutive 

baby boomers presented for routine care and underwent 

opt-out HCV screening from Sept 2013 through June 2014. 

Educational materials were distributed by ED staff to HCV Ab 

positive subjects. A LTC coordinator undertook follow-up phone 

calls, charity care applications for the uninsured and letter mailing 

for contact failures. Electronic Health Record was queried 

for demographics, ICD-9 codes for major morbidity domains 

and visit logs. LTC was defined as at least one HCV clinic 

visit post screening. Results: Over 4300 baby boomers were 

screened and 473 tested positive for HCV Ab (11%). Viral 

load was checked in 402 subjects and 336 patients had confirmed 

HCV viremia (84%). Among viremic patients, the mean 

age was 57.3 years (SD± 4.8), 70% were males and 61% 

were African Americans. Forty-two percent had public insurance, 

14% were uninsured and 82% had multi-morbidity (≥ 2 

morbidity domains). Substance abuse (35%), psychiatric disease 

(29%) and cirrhosis (31%) were highly prevalent. A total 

of 161 baby boomers (48%) were hospitalized and 221 (66%) 

utilized multi-specialty care within campus during follow-up. 

Median follow-up was 433 days (IQR 354 - 500). The median 

time between screening and the first HCV care visit was 81 

days (IQR 40 - 205). Overall, 121 baby boomers (36%) did 

not attend any visits on campus, 94 (29%) attended non-HCV 

care visits only and 117 (35%) attended HCV care visits at 

least once during follow-up. Lack of HCV clinic attendance 

had independent associations with Males (aOR 1.79, 95% 

CI 1.0 – 3.19), Uninsured Status (aOR 5.62, 95% CI 1.57- 

20.11), Hospitalization (aOR 0.52, 95% CI 0.29 – 0.93) and 

Primary Care Attendance in the university system (aOR 0.22, 

95% CI 0.10 – 0.43). Conclusions: In this interim analysis, 

approximately 1 of every 3 HCV-infected baby boomers was 

linked to a HCV treatment center following ED screening. LTC 

interventions in this setting should be robust in order to address 

significant lack of access to care, frequent competing medical 

priorities and meet the great need created by high-yield screening. 

Further research is warranted for optimal LTC practices. 

Disclosures: 

Ricardo A. Franco - Advisory Committees or Review Panels: Gilead; Grant/ 

Research Support: Gilead 

Michael Saag - Grant/Research Support: BMS, Gilead, Abbvie, Merck, ViiV, 

Janssen 

The following authors have nothing to disclose: Edgar T. Overton, Ashutosh 

Tamhane, Jordan M. Forsythe, Joel B. Rodgers, Deanne Guthrie, Suneetha Thogaripally, 

Anne Zinski, Michael Mugavero, James W. Galbraith 

1805 

Racial Differences in Socioeconomic Status (SES), Cirrhosis, 

and Treatment of Chronic Hepatitis C (CHC): Highest 

Treatment Rates in Caucasian Americans (CA), followed 

by Hispanic Americans (HA), African Americans (AFA) 

and Lowest in Asian Americans (AA) 

Philip Vutien 2,1 , Joseph K. Hoang 1 , Louis Brooks 3 , Mindie H. 


University Medical Center, Palo Alto, CA; 2 Rush University Medical 

Center, Chicago, IL; 3 Optuminsight, Eden Prairie, MN 

Background: In the USA, CHC disproportionately affects ethnic 

minorities with higher prevalences but their linkage to care 

remains understudied. Methods: We performed a retrospective 

study of a US adult cohort of 76,849 CHC patients identified 

via ICD9 query from OptumInsight’s Data Mart from 1/2009- 

12/2013. All patients had continuous medical and pharmacy 

coverage: 72,746 with commercial insurance and 4,103 with 

Medicaid. Results: The majority were male (61%) and older 

than 50 (62%). The proportions of patients with a BA/BS 

degree were highest in AA (28%), followed by CA (14%), HA 

(7%), and AFA (4%). Similarly, household income > 100,000 

USD were most frequently seen in AA (45%) and CA (36%) 

and less often in HA (20%) and AFA (15%). At presentation, 

HA were most likely to have cirrhosis compared to CA, AA, 

and AFA (35% vs. 31%, 30%, and 30%, respectively). AA 

were the least likely to have any psychiatric (8%) or medical 

comorbidity (54%) while CA were the most likely to have a 

psychiatric comorbidity (21%) and AFA to have a medical 

comorbidity (76%). Only 10% of all patients and 0.1% of 

Medicaid patients were prescribed anti-HCV treatment. AA 

had the lowest prescription rates at 7.8% overall and in each 

income category (Figure). In a multivariate analysis, CA was 

a significant predictor of anti-HCV treatment after adjustment 

for medical and psychiatric comorbidities, education, income, 

age, and cirrhosis with odds ratios of 1.52 for CA vs. AA, 

1.26 for CA vs. HA, and 1.18 for CA vs. BA. Conclusions: In a 

cohort of insured patients, ethnic minorities (HA, AFA, AA) had 

lower anti-HCV treatment rates than CA. Despite having fewer 

comorbidities and higher incomes and educational levels, AA 

had the lowest treatment rates. Future studies should aim to 

identify underlying ethnic-related barriers to anti-HCV treatment 

besides SES and comorbidities.


of CHC. Older age and receiving treatment were significant 

predictors of developing cirrhosis. 

Disclosures: 

Louis Brooks - Employment: Optum 





The following authors have nothing to disclose: Philip Vutien, Joseph K. Hoang 

1806 

Progression to liver cirrhosis in patients with chronic 

hepatitis C (CHC) in a large United States cohort: a natural 

history study 

Nghia H. Nguyen 2,1 , Joseph K. Hoang 1 , An K. Le 1 , Changqing 

Zhao 1,3 , Christine Y. Chang 1 , Richard H. Le 1 , Mingjuan Jin 1,4 , 

Alina Kutsenko 5 , Lee Ann Yasukawa 6 , Jian Q. Zhang 7 , Susan C. 

Weber 6 , Mindie H. Nguyen 1 ; 1 Division of Gastroenterology and 

Hepatology, Stanford University Medical Center, Palo Alto, CA; 

2 Medicine, University of California, San Diego, San Diego, CA; 

3 Department of Cirrhosis,Institute of Liver Disease, Shuguang Hospital, 

Shanghai, China; 4 Department of Epidemiology and Biostatistics, 

Zhejiang University School of Public Health, Hangzhou, 

China; 5 Department of Medicine, Stanford University Medical 

Center, Palo Alto, CA; 6 Center for Clinical Informatics, Stanford 

University School of Medicine, Palo Alto, CA; 7 Chinese Hospital, 


Purpose: Most CHC studies in the U.S. are based on estimates 

from simulation models. Our goal was to describe CHC disease 

progression to cirrhosis in a real-life cohort from 2 ethnically 

diverse U.S. centers. Methods: ICD-9 query identified 9207 

consecutive CHC patients in 2005-2014. Analysis was performed 

from data obtained via individual chart review in 7105 

confirmed CHC cases. Kaplan Meier method was performed 

to estimate cumulative incidence of cirrhosis. Cox regression 

was performed to identify predictors of disease progression 

Results: The majority of our patients were men (62%), had 

genotype 1 (69%) and mean age of 55.4±10.7. Treatment 

rate was low (13%) in our cohort with the majority of treated 

patients having received interferon-based therapy (58%). 

Crude incidence for development of cirrhosis was 353.7 per 

10,000 person-years. On subgroup analyses, men (vs women) 

had a significantly higher overall 5-year cumulative incidence 

of cirrhosis (p=0.009) and also when stratified by age (


12% self-reported daily alcohol use. Among 895 persons not 

treated for HCV, 181 developed ESLD and 30 developed HCC 

during a total follow-up of 7,896 person-years. Among 69 

persons who achieved an SVR, 5 developed ESLD and none 

developed HCC during a total follow-up of 437 person-years 

after completing HCV treatment. Among 98 persons who failed 

or discontinued HCV treatment, 11 developed HCC and 37 

developed ESLD during a total follow-up of 614 person-years. 

Compared with persons never treated for HCV, persons who 

failed treatment were more likely to develop ESLD (adjusted 

hazard ratio [a-HR]: 2.67, 95% confidence interval [CI] 1.82, 

3.91) and HCC (aHR: 4.22, CI: 1.88, 8.70). Persons who 

achieved SVR were less likely to develop ESLD (a-HR: 0.29, CI: 

0.09, 0.93). Conclusions: HCV RNA positive AN/AI persons 

failing to achieve SVR after IFN-based therapy are at increased 

risk of developing ESLD and HCC as compared to those never 

treated and should be prioritized for treatment with direct-acting 

antiviral agents. 

Disclosures: 

Lisa J. Townshend-Bulson - Grant/Research Support: Gilead 

Chriss E. Homan - Grant/Research Support: Gilead 

James E. Gove - Grant/Research Support: Gilead Sciences 

Julia N. Plotnik - Grant/Research Support: Gilead 

Hannah G. Espera - Grant/Research Support: Gilead 

Annette Hewitt - Grant/Research Support: gilead 

Youssef Barbour - Grant/Research Support: Gilead 

The following authors have nothing to disclose: Dana J. Bruden, Brian J. McMahon, 

Stephen Livingston, Brenna Simons-Petrusa, Susan Negus, Mary Snowball, 

Prabhu P. Gounder 

1808 

Benefits of HCV eradication in compensated cirrhotic 

patients extend beyond liver-related complications: 

results from the ANRS CO12 CirVir prospective cohort. 

Pierre Nahon 1 , Valerie Bourcier 1 , Richard Layese 2 , Ventzislava 

Petrov-Sanchez 3 , Dominique Guyader 4 , Sebastien Dharancy 5 , 

Hélène Fontaine 6 , Vincent Thibault 7 , Vincent Leroy 8 , Stanislas 

Pol 6 , Françoise Roudot-Thoraval 2 ; 1 Hôpital Jean Verdier, Bondy, 

France; 2 Hôpital Henri Mondor, Créteil, France; 3 ANRS, Paris, 

France; 4 CHU Pontchaillou, Rennes, France; 5 Hôpital Huriez, Lille, 

France; 6 Hôpital Cochin, Paris, France; 7 GH Pitié-Salepêtrière, 

Paris, France; 8 Hôpital Michallon, Grenoble, France 

Backround and aims: The objective was to prospectively assess 

the benefits of sustained viral response (SVR) on the incidence 

of liver-related and extra-hepatic complications in French 

patients with compensated HCV cirrhosis using competing 

risk analyses. Methods: Patients with the following inclusion 

criteria were enrolled in 35 French centres: a) biopsy-proven 

HCV cirrhosis; b) Child-Pugh A; c) absence of previous liver 

complications. Patients were prospectively followed-up every 

6 months. Sustained virological response (SVR) was defined 

as undetectable HCV RNA at the end of a 12-week untreated 

follow-up period. A given event was arbitrarily considered as 

occurring in a patient who achieved SVR if it was recorded at 

least one year after successful treatment completion. Results: 

A total of 1323 patients were enrolled between March 2006 

and June 2012 [age 55, men 63.5%, Genotype 1: 66%, 

2: 6 %, 3: 17%, 4: 10%, 5-6: 1%]. At end-point in January 

2015 (median follow-up of 51 months), the number (rates) 

of HCV patients with a negative viral load was 653 (51%) 

corresponding to SVR in 491 patients (40%) while the remaining 

162 HCV negative patients were still undergoing antiviral 

treatment (mostly based on second generation DAAs). Primary 

liver cancer (PLC) was diagnosed in 168 (162 hepatocellular 

carcinoma (HCC) and 6 cholangiocarcinomas) among which 

17 were developed in SVR patients (P


Liver histological examination revealed that 7 patients had 

fibrosis grade F0, 149 had F1, 87 had F2, 58 had F3, 16 had 

F4. Both serum levels of Fuc-Hpt and M2BP increased along 

the progression of liver fibrosis stage. The median of serum 

Fuc-Hpt levels in patients with fibrosis grade F0-1, F2, F3, and 

F4 were 433, 592, 1022, and 1790 U/ml respectively. The 

M2BP median with F0-1, F2, F3, and F4 were 1261, 2276, 

2740, and 4044 ng/ml. We evaluated the diagnostic ability 

of aspartate aminotransferase to platelet ratio index (APRI), 

FIB4 index, Fuc-Hpt and M2BP in liver fibrosis grade F ≥ 2, F ≥ 

3 and F4 by ROC analysis. The AUROC of Fuc-Hpt in F ≥ 2, F 

≥ 3 and F4 were 0.677, 0.738 and 0.813 respectively. M2BP 

were 0.747, 0.734 and 0.797. Most AUROC of Fuc-Hpt and 

M2BP were over 0.7 in all fibrotic stages, suggesting that both 

Fuc-Hpt and M2BP could be applicable for evaluating liver 

fibrosis. However, these markers did not exceed the values of 

known fibrotic index such as APRI or FIB4 index. To establish 

better liver fibrosis markers, we evaluated the combination of 

known marker with new marker. To evaluate for fibrosis grade 

F ≥ 2, the AUROC of FIB4 index adding M2BP was most diagnosable; 

0.800. To evaluate for fibrosis grade F ≥ 3 and F4, 

FIB4 index adding Fuc-Hpt is most diagnosable; 0.841 and 

0.913. We examined to identify the factors associated with the 

progression of liver fibrosis grade F ≥ 2 and F ≥ 3. In multivariate 

analysis revealed that M2BP, FIB4 index and AFP were the 

independent factors associated with liver fibrosis grade F ≥ 2. 

And Fuc-Hpt, FIB4 index and AFP were the independent factors 

in F ≥ 3. Finally we evaluated the cumulative hepatocellular 

carcinoma (HCC) incidence rate. The follow up period after 

liver biopsy is 1148 ± 913 days. HCC was occurred in 19 

patients during follow up periods. The cumulative incidence of 

HCC was significantly higher in the patients with high serum 

levels of Fuc-Hpt (> 559 U/ml; median) or high M2BP (> 1847 

ng/ml; median) than in patients with low levels of them by log 

rank test. Conclusion: Both Fuc-Hpt and M2BP could be useful 

biomarkers to evaluate liver fibrosis in patients with chronic 

hepatitis C. 

Disclosures: 






The following authors have nothing to disclose: Seiichi Tawara, Tomohide 

Tatsumi, Sadaharu Iio, Ichizo Kobayashi, Shigeki Suemura, Atsuo Takigawa, 

Tadashi Kegasawa, Takahiro Suda, Teppei Yoshioka, Yoshiki Onishi, Akira 

Nishio, Satoshi Aono, Minoru Shigekawa, Ryotaro Sakamori, Naoki Hiramatsu, 

Eiji Miyoshi 

1810 

An Integrated Linkage to Care Model to Improve Patient 

Engagement and Retention in HCV Specialty Care 

Catelyn Coyle 1 , Kendra Viner 2 ; 1 National Nursing Centers Consortium, 

Philadelphia, PA; 2 Disease Control, Philadelphia Department 

of Public Health, Philadelphia, PA 

Purpose: To describe a successful linkage to care model piloted 

in a network of community health centers in Philadelphia, PA. 

Methods: Hepatitis C (HCV) patients are lost at each stage 

of the HCV continuum of care. In September 2013, National 

Nursing Centers Consortium implemented a linkage model 

to usher HCV-positive patients receiving care in one of five 

Philadelphia based community health centers into specialty 

care. Eligible patients were those testing HCV-antibody positive 

after October 1, 2012. The model included a linkage to care 

coordinator, EMR modifications, and the following services to 

help patients overcome barriers: escorts to appointments, transportation 

assistance (including tokens), appointment scheduling, 

coordination with sub-specialty offices, and connection to 

social services. EMR changes, including prompts, result summary 

flow sheets, and weekly reports, ensured that patients 

were followed through the entire care cascade. Patients with 

missed appointments were contacted to reschedule and identify 

barriers. Calls to emergency contacts and site visits were made 

to reach any patients lost to care. Results: During September 

2013 - February 2015, 4,893 patients were HCV tested. Of 

these, 601 (12%) had a positive HCV-antibody test, 584 (97%) 

received an HCV-RNA test, and 406 (69%) were identified 

with current HCV infection. Of the 406, 366 (90%) received 

their HCV-RNA positive result, 327 (81%) were referred to an 

HCV care provider, and 231 (57%) were seen by the referred 

HCV care provider. During the 18-month study period, testing 

and linkage to care rates ranged across the five testing sites: 

97%-100% for HCV-antibody-positive patients receiving HCV- 

RNA testing, 53%-73% for HCV-RNA positivity, 83%-100% 

for receipt of a positive HCV-RNA test result, 67%-89% for 

referral to HCV care, and 37%-72% for HCV care provider 

visits. Lack of insurance, transportation, weather-appropriate 

clothing, and being signed out of shelters were identified as 

the most common social barriers to care. During the 11 months 

before the addition of a linkage to care coordinator (October 

2012-August 2013) and the 11 months afterwards (September 

2013-July 2014), the number of HCV-infected patients receiving 

their positive HCV-RNA result rose by 68%, patient referrals 

by 49%, and patients seen for HCV care by 29%. Conclusion: 

Intensive linkage services can increase the number of patients 

who successfully navigate the HCV treatment cascade. The linkage 

to care coordinator is an important position that acts as a 

trusted intermediary for patients being linked to care. 

Disclosures: 

Catelyn Coyle - Advisory Committees or Review Panels: Gilead Sciences 

The following authors have nothing to disclose: Kendra Viner 

1811 

Increasing Hepatitis C Virus (HCV) Screening and Linkage 

to Care in a Large Integrated Health System 

Carla V. Rodriguez 1 , Kevin Rubenstein 1 , Haihong Hu 1 , Benjamin 

P. Linas 2,3 , Michael A. Horberg 1 ; 1 Mid-Atlantic Permanente 

Research Institute, Kaiser Permanente, Rockville, MD; 2 HIV Epidemiology 

and Outcomes Research Unit, Section of Infectious Diseases, 

Boston Medical Center, Boston, MA; 3 Epidemiology, Boston 

University School of Public Health, Boston, MA 

Objective: To describe the HCV cascade of care from screening 

to linkage in Kaiser Permanente, Mid-Atlantic States (KPMAS) 

relative to the 2013 release the U.S. Preventative Services Task 

Force “birth cohort” (born 1945-1965) screening recommendations. 

Methods: Cohort study among patients in KPMAS with 

≥8 months of enrollment from 1/1/2003-12/31/2014 and 

≥1 clinical visit. Birth cohort, IDU, MSM, sex, race, clinic location, 

income, elevated ALT (2 consecutive ALT >60 U/L), HIV 

and HBV status were characteristics of interest. We estimated 

the annual screening rate as the number antibody (Ab) tested 

per persons enrolled each year. We used survival methods to 

describe factors associated with time to Ab testing, stratified by 

location; and logistic regression to identify predictors of GI/ID 

referral. Among Ab+, we describe the cumulative probability 

and predictors of confirmatory RNA or genotype testing, GI/ID 

referral, and HCV treatment. Results: We observed 665,345 

patients over an 11 year period. The annual screening rate 

increased steadily from 23.6 to 70.8 per 1,000 person-years 

from 2004 to 2014; with the sharpest increase after 2013. A 

total of 123,572 (18.6%) KPMAS members were screened for 

HCV. Screening among the birth cohort was lower than non-co-


hort members. However, the gap began to shrink during later 

years of study. Across all locations, screening was increased 

among drug users, HBV and HIV status, and elevated ALT. 

Screening was lower among men. Among Ab+, 84% were 

RNA tested. In an adjusted model, we found no differences 

in confirmatory testing by location, MSM, race, drug use, HIV 

or HBV status. Elevated ALT (aHR=1.3; ref=ALT


for (1) appropriate resource allocation to meet the needs of 

the affected patients and (2) strategic deployment of antiviral 

therapy to mitigate the current and future burden of HCV. 

Disclosures: 



















The following authors have nothing to disclose: Henri Cournand 

1814 

Patients with Hepatitis C (HCV) Advanced Cirrhosis and 

the rs738409 GC/GG Genotype of Platatin-Like Phosphatase 

Domain Containing (PNPLA-3) are at Risk for 

Further Decompensation Despite Direct-Acting Antiviral 

(DAA) Treatment 

Winston Dunn 1 , Anusha Vittal 1 , Melissa Whitener 1 , Jie Zhao 1 , 

Brian Bridges 1 , Shweta Chakraborty 1 , K. James Kallail 2 , Prashant 

K. Pandya 3 , Chuanghong Wu 4,1 , Ryan Taylor 1 , Jody C. Olson 1 , 

Richard Gilroy 1 , Mojtaba S. Olyaee 1 , Timothy Schmitt 1 , Steven 

A. Weinman 1 ; 1 The University of Kansas Medical Center, Kansas 

City, KS; 2 Office of Research, The University of Kansas School of 

Medicine-Wichita, Wichita, KS; 3 Kansas City VA Medical Center, 

Kansas City, MO; 4 Shekou People’s hospital, Shenzhen, China 

HCV patients with compensated cirrhosis generally do well 

after successful virological cure, but approximately 1/3 of 

patients with decompensated cirrhosis experience further 

decompensation or develop hepatocellular carcinoma (HCC). 

The PNPLA3 genotype is correlated with fibrosis progression 

from HCV. We tested the hypothesis that the PNPLA3 genotype 

can predict recovery versus further decompensation in 

advanced cirrhosis after HCV is cured. We studied 27 patients 

with advanced HCV cirrhosis who were treated with an interferon-free 

DAA-based treatment regimen. We excluded patients 

with HCC, previous transplantation, and censored patients 

upon transplantation. Nine patients had bilirubin ≥2 mg/dl or 

INR ≥2 before DAA treatment. All remaining patients had clinical 

decompensation. Eight were listed for transplantation. The 

median follow-up was 166 days. The primary outcome was 

a change in Model for End Stage Liver Disease (MELD) from 

before DAA treatment. Figure 1 shows a comparison of before 

DAA treatment to the end of follow-up. Two patients, both with 

the GC genotype, died of sepsis and variceal bleeding, and 

one patient each with GC and CC genotypes underwent liver 

transplantation. Using a random coefficient model that considers 

repeated measurement of MELD and adjusting for baseline 

MELD and duration since treatment, patients with the CC allele 

dropped an average of 2.5 (95% confidence interval, 1.4 - 

3.6) MELD points compared to patients with the GC/GG allele 

(p = 0.0001). The effect remained significant after adjusting 

for age, gender, diabetes, and BMI (p = 0.02).This prospective 

cohort study showed a statistically significant and clinically 

meaningful difference in which patients with HCV advanced 

cirrhosis and an unfavorable PNPLA3 genotype are at risk 

for further decompensation despite virological cure. PNPLA3 

genotyping may be clinically useful to better target patients for 

liver transplant evaluation. 

Disclosures: 

Prashant K. Pandya - Grant/Research Support: Gilead, Merck; Speaking and 

Teaching: Genentech, AbbVie, Vertex 




The following authors have nothing to disclose: Winston Dunn, Anusha Vittal, 

Melissa Whitener, Jie Zhao, Brian Bridges, Shweta Chakraborty, K. James 

Kallail, Chuanghong Wu, Ryan Taylor, Mojtaba S. Olyaee, Timothy Schmitt 

1815 

Natural History of Decompensated Cirrhosis after 

Direct-Acting Antivirals (DAA) Treatment of Hepatitis C 

(HCV) 

Winston Dunn 1 , Melissa Whitener 1 , Shweta Chakraborty 1 , Anusha 

Vittal 1 , Prashant K. Pandya 2 , K. James Kallail 3 , Brian Bridges 1 , 

Chuanghong Wu 4 , Ryan Taylor 1 , Jody C. Olson 1 , Richard Gilroy 1 , 

Timothy Schmitt 1 , Mojtaba S. Olyaee 1 , Steven A. Weinman 1 ; 1 The 

University of Kansas Medical Center, Kansas City, KS; 2 Kansas 

City VA Medical Center, Kansas City, MO; 3 Office of Research, 

The University of Kansas School of Medicine-Wichita, Wichita, KS; 

4 Shekou People’s hospital, Shekou, China 

With the recent approval of newer DAA, more than 95% of 

patients with HCV infections can achieve a Sustained Virological 

Response (SVR). Although patients with compensated 

cirrhosis generally do well after viral clearance, the outcome 

for decompensated cirrhosis after SVR is not well defined. This 

study reports the outcomes of a cohort of decompensated HCV 

cirrhosis patients undergoing interferon free DAA-treatment in 

an academic transplant center. We included 96 patients in 

total all of whom had one or more indications of advanced disease 

such as hepatocellular carcinoma (HCC, n=21), clinical 

decompensation defined as hepatic encephalopathy, ascites, 

or variceal bleeding (n=81) or biochemical decompensation 

(bilirubin ≥2 mg/dl or INR ≥1.7, n=47). The median follow-up 

was 181 days. Six patients received liver transplantation and 

3 died. Three patients had to discontinue treatment due to 

adverse events. SVR occurred in 33/45 eligible (adequate


follow-up time) patients. Post treatment MELD score changes 

showed considerable heterogeneity with SVR patients showing 

an average MELD decrease of 2.0 (95% confidence interval 

0.9 - 3.1, p=0.0003) MELD points compared to patients who 

failed SVR. Even within the SVR group, 33% of the patients 

had post treatment increases in MELD score while 63% had a 

decrease. Four patients had refractory ascites requiring paracentesis 

prior to treatment and none required paracentesis after 

SVR. One patient developed refractory ascites after treatment. 

Four patients had hepatic encephalopathy (PSE) related hospital 

admission prior to treatment but only 1 had further PSE 

related admissions after treatment. Three new patients developed 

PSE related admission for the first time after treatment, 

and one patient developed HCC after SVR. In summary, 3/4 of 

decompensated HCV cirrhotics were able to achieve SVR with 

approximately 2/3 of the SVR patients achieving decreased 

MELD score. Most patients with refractory symptoms had clinical 

benefits such as reduced paracentesis and encephalopathy 

related admissions. Improvement is not universal and patients 

should be carefully monitored. DAA treatment for advanced 

cirrhosis is not a substitution for transplant. For those patients 

who did not achieve SVR, deterioration occurred in half. HCV 

treatment is thus indicated, even in the most advanced cases 

and a significant proportion of these patients may improve to 

the point of not requiring liver transplantation. 

Disclosures: 

Prashant K. Pandya - Grant/Research Support: Gilead, Merck; Speaking and 

Teaching: Genentech, AbbVie, Vertex 




The following authors have nothing to disclose: Winston Dunn, Melissa Whitener, 

Shweta Chakraborty, Anusha Vittal, K. James Kallail, Brian Bridges, Chuanghong 

Wu, Ryan Taylor, Timothy Schmitt, Mojtaba S. Olyaee 

1816 

Low HCV Treatment for Chronic Hepatitis C (CHC) 

Patients in Pre-Protease Inhibitor (PI) and Post-PI/Pre-Direct 

Acting Antiviral (DAA) Eras Compared to Post-DAA 

Era, Especially in African Americans: Analysis of A 

Large Real-World Cohort of 7105 Patients 

Peter T. Nguyen 2,1 , Nghia H. Nguyen 3,1 , Joseph K. Hoang 1 , 

Changqing Zhao 1,4 , An K. Le 1 , Christine Y. Chang 1 , Richard H. 

Le 1 , Michael D. Nguyen 1 , Mingjuan Jin 5,1 , Susan C. Weber 6 , Lee 

Ann Yasukawa 6 , Mindie H. Nguyen 1 ; 1 Division of Gastroenterology 

and Hepatology, Stanford University Medical Center, Palo 

Alto, CA; 2 University of Texas Medical Branch, Galveston, TX; 


Diego, CA; 4 Department of Cirrhosis, Institute of Liver Disease, 

Shuguang Hospital, Shanghai, China; 5 Department of Epidemiology 

and Biostatistics, School of Public Health, Zhejiang University, 

Hangzhou, China; 6 Center for Clinical Informatics, Stanford University 

School of Medicine, Palo Alto, CA 

Background & Aim: Utilization of PEG IFN+RBV (P/R) is generally 

low for CHC due to adverse events (AE) and low SVR. 

More efficacious triple P/R+PI therapy was approved in 2011 

but treatment rate may also remain low due to AE. Our goal 

was to measure HCV treatment rates in a large ethnically 

diverse U.S. single-center cohort and changes from 2005- 

2014. Methods: ICD-9 query identified 9207 patients and 

individual charts were reviewed and confirmed HCV diagnosis 

in 7105 patients. Of these, 929 received anti-HCV treatment 

(P or P/R-based DAA-based). Results: Overall, treatment rate 

was low (Figure). From 2005-2013, the proportion of treated 

patients per all CHC patients with at least one clinical encounter 

ranged 1.0-3.7% and 3.7-14.3% per new CHC patients. 

Treatment rates per new HCV patients from 2005-07, 2008- 

10, 2011-13, and 2014 were significantly different: 4.4%, 

11.5%, 9.3%, and 46.5% (p


E (clinical illness, with anti-HEV IgM +ve) a year ago, when 

their unit was affected by an outbreak (n=67), (B) other soldiers 

in the unit who did not have symptomatic hepatitis (n=133), 

and (C) soldiers in another unit located elsewhere in the city, 

which was not affected by the outbreak (n=196). All sera were 

tested for anti-HEV IgG using 3 commercial assays (Wantai 

[W], China; DSI [D], Italy; and MP Biomedical [M], Singapore); 

the manufacturers’ protocols were followed. Results: In 

each group, assay W had the highest sensitivity (Table). After 

excluding indeterminate results, which were most common with 

assay M, the 3 assays showed positive concordant results in 

55, 16 and 15 sera, and negative concordant results in 2, 75 

and 133 sera, in groups A, B and C, respectively. Discordant 

results were found in 10, 30 and 36 sera. Using positive results 

in ≥2 assays as the criterion-standard, the sensitivity rates of 

assays W, D and M were 100%, 100%, and 87%, respectively 

in group A; 100%, 95% and 47% in group B; and 97%, 

94% and 58% in group C. Similarly, using negative results 

in ≥2 assays as the criterion-standard, the specificity rates of 

assays W, D and M were 75%, 75%, and 100%, respectively 

in group A; 96%, 99% and 95% in group B; and 92%, 100% 

and 95% in group C. Overall assays W and D had good concordance 

(97%, 95% and 91% in A, B and C). Conclusion: Of 

the three assays, W and D had a higher sensitivity and high 

degree of concordance with each other. In comparison, assay 

M was less sensitive and had less concordance with the other 

two assays. Also, assay M had more indeterminate test results. 

These data indicate that assays W and D may be preferred 

over assay M for anti-HEV seroprevalence studies. 

Results with the three serological assays 

Indet. = Indeterminate 

Disclosures: 

The following authors have nothing to disclose: Rakesh Aggarwal, Amit Goel, 

Gurdeep Singh 

1818 

Hepatitis C Virus (HCV) Prevalence among People who 

Inject Drugs (PWIDs) in Switzerland 

Francesco Negro 1 , Philip Bruggmann 2 , Sarah Blach 3 , Pierre 

Deltenre 4 , Jan Fehr 5 , Roger Kouyos 5 , Daniel Lavanchy 6 , Beat Mullhaupt 

7 , Homie Razavi 3 , Patrick Schmid 8 , David Semela 9 , Martin 

Stoeckle 10 , Andri Rauch 11 ; 1 Gastroenterology, Hepatology and 

Clinical pathology, University Hospitals, Geneva-14, Switzerland; 

2 ARUD Center for addiction medicine, Zurich, Switzerland; 3 Center 

for Disease Analysis, Lousville, CO; 4 Gastroenterology and 

hepatology, University Hospital, Lausanne, Switzerland; 5 Infectious 

Diseases & Hospital Epidemiology, University Hospital, Zurich, 

Switzerland; 6 Consultant, Denges, Switzerland; 7 Gastroenterology 

and hepatology, University Hospital, Zurich, Switzerland; 

8 Infectious diseases, Cantonal Hospital, St Gallen, Switzerland; 

9 Gastroenterology, Cantonal Hospital, St. Gallen, Switzerland; 

10 Infectious Diseases & Hospital Epidemiology, University Hospital, 

Basel, Switzerland; 11 Infectious diseases, University Hospital, 

Berne, Switzerland 

Background - In Switzerland, HCV among PWIDs has been 

decreasing due to active harm reduction efforts and an aging 

population (average age – 44 yrs). Expert consensus and estimates 

from the Swiss Federal Office of Public Health suggest 

that there are between 8,000 and 12,000 active PWIDs in 

Switzerland, and that 42% (27%-58%) of PWIDs are HCV 

infected. In addition, an estimated 17,000 – 25,700 individuals 

were enrolled in opioid substitution therapy (OST) and 

1,598 in heroin substitution therapy (HeGeBe) with 300,000 

syringes distributed monthly in 2012. Among individuals on 

OST and HeGeBe, an estimated 27.4% and 54%, respectively, 

continued to inject while on treatment. Understanding HCV 

transmission dynamics among high-risk populations requires 

robust epidemiological data and country-specific mathematical 

modeling to assess the potential impact of new HCV treatment 

strategies. Recent therapeutic advances promise greater convenience 

(oral therapies) with higher efficacy (>90% sustained 

viral response) and shorter duration of treatment. Methods - 

HCV transmission was modeled using cohorts to track HCV 

incidence and prevalence among PWIDs in the general population, 

as well as active PWIDs enrolled in OST and/or needle 

exchange programs (NEP). Model assumptions were derived 

from published literature and expert consensus. The relative 

impact of increasing treatment among PWIDs was considered, 

including the annual number needed to treat in order to reduce 

the HCV-infected PWID population by 2030. Results - If the 

current transmission paradigm continues, there are projected 

to be 3,150 HCV infected PWIDs in 2030. Annually treating 

45 HCV-infected PWIDs (1% of HCV-infected PWID population 

in 2014) resulted in an 11% reduction in HCV-infected 

PWIDs by 2030, while annual treatment of 200 PWIDs (4.5% 

of 2014 population) resulted in a reduction of over 50% by 

2030. Treating 322 PWIDs annually (7.5% of 2014 population) 

resulted in a >90% reduction in HCV-infected PWIDs by 

2030. Targeting treatment to PWIDs engaged in OST and NEP 

provided the greatest reduction in prevalence for the number of 

individuals treated. To achieve a 1-person reduction in overall 

prevalence by 2030, it was necessary to treat 1.6 PWIDs in 

OST/NEP as compared with 3.4 PWIDs in the general population. 

Conclusions - Treating a relatively small number of PWIDs 

results in substantial decreases in the HCV infected PWID population 

by 2030. Additionally, the impact of treatment is higher 

among PWIDs engaged in harm reduction programs. These 

data support implementation of a screening and treatment strategy 

among PWIDs, and particularly among PWIDs engaged 

in OST and NEP. 

Disclosures: 



Philip Bruggmann - Advisory Committees or Review Panels: Merck, Gilead, BMS, 

Abbvie, BMS; Grant/Research Support: BMS, Merck, Janssen, Gilead, Abbvie; 


Sarah Blach - Employment: Center for Disease Analysis 

Pierre Deltenre - Consulting: Abbvie, Gilead, BMS; Grant/Research Support: 

Abbvie, Gilead, BMS, Janssen, MSD; Speaking and Teaching: Abbvie, Gilead, 

BMS 

Beat Mullhaupt - Consulting: MSD, Novartis, MSD, Janssen; Grant/Research 

Support: Bayer, Gillead 

Homie Razavi - Management Position: Center for Disease Analysis 

Andri Rauch - Advisory Committees or Review Panels: Gilead Sciences, Abbvie, 

MSD, Janssen Cilag 

The following authors have nothing to disclose: Jan Fehr, Roger Kouyos, Daniel 

Lavanchy, Patrick Schmid, David Semela, Martin Stoeckle


1819 

The clinical profiles of Japanese patients with L31M and 

Y93H variants in NS5A region of hepatitis C virus 

Kei Ogata 1 , Tatsuya Ide 1 , Teruko Arinaga-Hino 1 , Ichiro Miyajima 

1 , Reiichiro Kuwahara 1 , Keisuke Amano 1 , Takumi Kawaguchi 

1 , Toru Nakamura 1 , Masahito Nakano 1 , Hironori Koga 1 , Ryoko 

Kuromatsu 1 , Yuichiro Eguchi 2 , Masaru Harada 3 , Takuji Torimura 1 ; 

1 Division of Gastroenterology, Department of Medicine, Kurume 

Univ.School of Medicine, Kurume, Japan; 2 Department of Internal 

Medicine, Faculty of Medicine, Saga University, Division of Hepatology, 

Saga, Japan; 3 The Third Department of Internal Medicine, 

School of Medicine, University of Occupational and Environmental 

Health, Kitakyusyu, Japan 

Background and aims: Therapy for hepatitis C patients has 

been developed, and several direct antiviral agents (DAAs) 

have been approved clinically. The NS5A replication complex 

inhibitor is one of the important DAAs. It is known that 

resistance-associated variants (RAVs) naturally occur at NS5A 

positions L31 and Y93. Since the clinical background and 

profiles are still unclear, we investigated RAVs by direct 

sequencing and cycleave methods. Patients and method: 

Patients with genotype 1 and who were treatment naive for 

NS5A inhibitors were selected. We measured the Y93 positions 

in 1,204 patients and L31 positions in 1,039 patients 

by direct sequencing. We also measured the Y93 positions in 

1,039 patients by the cycleave method (PLoS One. 2014, Nov 

14:e112647). Among them, 520 patients of age, sex, with or 

without cirrhosis, HCV RNA level, ALT, platelet count, albumin, 

gamma-GTP, type IV collagen, prothrombin time and αfetoprotein, 

Il28B were also analyzed as clinical features. Results: 

In the 1,204 patients, the Y93H variant was detected in 165 

patients (13.7%). Among these 165 patients, 150 patients 

were measured by the cycleave method : 25.3% (38/150) of 

the patients had 100% of the Y93H variant, and the remaining 

patients had a mixed Y93H and Y93 wild type (Y93H variant, 

0-20% ; 52 patients (34.6%), 21-40% ; 18 patients (12.0%), 

41-60% ; 15 patients (10.0%), and 61-99% ; 27 patients 

(18.0%). Patients with the Y93H variant had a significantly 

higher HCV RNA level (6.4 ± 0.3 vs. 6.0 ± 0.7 log IU / ml, 

respectively, p = 0.0016), lower platelet count (106 ± 45 vs. 

134 ± 66 x10 3 /ml, respectively, p = 0.0048), and an older 

age (72.1 ± 7.9 vs. 68.6 ± 8.9, yrs, respectively, p = 0.011) 

than patients with the Y93 wild type. On the other hand, the 

L31M variant was detected in 4.5% of patients (47/1,039). 

Patients with the L31M variant had a significantly lower HCV 

RNA level (5.5 ± 0.8 vs. 6.0 ± 0.7, log IU/ml, respectively, 

p = 0.0021), higher gamma-GTP (75.3 ± 79 vs. 50.2 ± 57, 

IU/L, respectively, p = 0.0074), and a higher proportion of 

males (10.1vs. 2.7%, respectively) than patients with the L31 

wild - type. Conclusion: We demonstrated the clinical features 

of naturally occurring L31 and Y93 variants. HCV RNA levels 

are closely related to RAVs. Although careful attention should 

be paid on treatment with NS5A inhibitors, the interaction 

among RAVs, these clinical features, and treatment efficacy 

should be investigated. 

Disclosures: 

The following authors have nothing to disclose: Kei Ogata, Tatsuya Ide, Teruko 

Arinaga-Hino, Ichiro Miyajima, Reiichiro Kuwahara, Keisuke Amano, Takumi 

Kawaguchi, Toru Nakamura, Masahito Nakano, Hironori Koga, Ryoko Kuromatsu, 

Yuichiro Eguchi, Masaru Harada, Takuji Torimura 

1820 

Hepatitis C Virus Genotype Analysis in Chronic Hepatitis 

Patients and Individuals with Spontaneous Virus Clearance 

Using a Newly Developed Serotyping Assay 

Ruifeng Yang, Huiying Rao, Lai Wei; Peking University People’s 

Hospital, Peking University Hepatology Institute, Beijing, China 

Objective We developed and evaluated the performance of 

an HCV serotyping assay in chronic hepatitis C(CHC) patients, 

and detected the genotypes in people with spontaneous viral 

clearance(SVC). Methods The novel serotyping assay was 

developed based on competitive ELISA principle. 997 CHC 

patients were enrolled in a nationwide cross-sectional study. 

Their samples were genotyped originally using the LiPA 2.0 

assay. For samples with inconsistent results yielded by the serotyping 

assay, sequences of the NS5B or 5’-UTR were analyzed 

to confirm the genotype results. In these patients, 190 patients 

achieved sustained virological response(SVR). The paired serotyping 

results were compared before and after treatment. Moreover, 

326 serum samples from a long-term follow-up cohort 

were also serotyped, among whom 66 were from SVC individuals, 

and the remaining 260 had persistent HCV RNA. All 

patients provided informed consent. Results There were 958 

out of 997 samples from CHC patients which yielded both 

genotyping and serotyping results. The overall consistency 

between the genotyping and serotyping assays was 90.19% 

(864/958). The specificity and sensitivity were 99.74% and 

88.77% for genotype 1, 96.97% and 93.97% for genotype 

2, 100% and 100% for mixed genotype 1/2, and 94.15% 

and 89.61% for non-genotype 1 or 2. In 190 SVR patients, no 

statistical difference existed in the serotype results after therapy(P=0.64). 

In the 326 follow-up patients, there was no difference 

in the proportion of genotypes between the groups with or 

without SVC(P=0.23, Tab 1). Conclusions The novel serotyping 

assay is sensitive and specific compared with the molecular 

genotyping assays. Using this assay, genotype distribution in 

SVC patients can also be revealed, which is similar with that 

in CHC patients. Genotype might not play an important role 

in SVC. 

Characteristics of the individuals with and without SVC 

Disclosures: 



The following authors have nothing to disclose: Ruifeng Yang, Huiying Rao


1821 

Mathematical model for early detection of hepatocellular 

carcinoma on top of HCV infection 

Abdelrahman Zekri, Amira S. Youssef, Yasser M. Bakr, Reham 

Mohamed Gabr, Ola Ahmed, Mohamed Abouelhoda; Virology 

and immunology Unit, Cancer Biology, National Cancer Institute, 

Cairo, Egypt 

Background: We investigated whether combinations of serum 

Intercellular Adhesion Molecules (sICAM-1), Beta catenin 

(β-catenin), Interleukin 8 (IL-8), Proteasome and soluble Tumor 

Necrosis Factor Receptor II (sTNF-RII) or subsets of them with 

AFP, used with logistic disease predictor models, could facilitate 

the early detection of hepatocellular carcinoma (HCC). 

Methods: Serum levels of IL-8, sICAM-1, sTNF-RII, Proteasome 

and β-catenin were measured in 479 subjects (192 

HCC associated with HCV infection; 96 HCV related liver 

cirrhosis (LC); 96 chronic hepatitis C (CHC) and 95 healthy 

controls). The R package and different modules for binary 

and multi-class classifiers based on generalized linear models 

were used to model the data. ROC curve analysis was 

used to find the best cutoffs differentiating among different 

groups. Results: We revealed mathematical models, based on 

binary classifier, made up of unique panel of biomarkers that 

improved the individual performance of AFP for the discrimination 

of HCC from benign liver disease. For the discrimination 

of HCC group from LC group, using a mathematical model 

[-1.133(E+01) +7.380* Proteasome + (1.081E-03)*sICAM-1+ 

(2.574E-01)*β-catenin + (1.597E-02)*AFP] with cutoff 0.6552 

has achieved 98.8% specificity and 89.1% sensitivity. For the 

discrimination of HCC group from CHC group, a mathematical 

model [-1.040*(e+01) + (1.416e+00)*Proteasome + 

(2.024e-03)*IL-8+ (4.096e-03)*sICAM-1+ (4.251e-04)*sT- 

NF-RII+ (2.567e-01)*β-catenin + (2.442e-02)*AFP] with cutoff 

0.744 has achieved 96.8% specificity and 89.7% sensitivity. 

Also, we have derived an algorithm for resolving the multi-class 

classification problem by using three successive mathematical 

models. First, use the mathematical model [-1.10E+01+6.83* 

Proteasome + (1.29E-03)*sTNF—RII + (2.83E-01)*AFP] 

with cutoff 0.764 to discriminate healthy controls from disease 

cases. For disease cases, discriminate HCC cases from 

HCV cases (cases with LC and CHC) using the mathematical 

model [-1.191(E+01) + (3.222E+00)* Proteasome + 

(3.813E-04)*IL-8 + (1.518E-03)*sICAM-1 + (6.481E-04)*sT- 

NF-RII + (2.906E-01)* β-Catenin + (1.931e-02)*AFP] with 

cutoff 0.712. If the cases were HCV, proceed to discriminate 

LC cases from CHC cases using the mathematical model 

[-1.016e+01 + (-4.487e+00)*Proteasome + (2.086e-03)*sI- 

CAM-1 + (1.858e-03)*sTNF-RII+ (-2.984e-01)* β-Catenin + 

(2.169e-02)*AFP] with cutoff 0.582. Conclusion: Our proposed 

mathematical models can differentiate not only between 

HCC and other studied groups but also between all studied 

groups in non-invasive, inexpensive and rapid manner. Key 

words: Hepatocellular carcinoma, AFP, sICAM-1, β-catenin, 

IL-8, sTNF-RII, Proteasome. 

Disclosures: 

The following authors have nothing to disclose: Abdelrahman Zekri, Amira 

S. Youssef, Yasser M. Bakr, Reham Mohamed Gabr, Ola Ahmed, Mohamed 

Abouelhoda 

1822 

Hepatitis C Seropositivity is not as high as Western 

Countries in the Population Born Between 1945-1965 

According to Turkey Nationwide Hepatitis Prevalence 

Study (TURKHEP 2010) 

Osman C. Ozdogan 1 , Nurdan Tozun 2 , Sabahattin Kaymakoglu 3 , 

Ramazan Idilman 4 , Zeki Karasu 5 , Ulus S. Akarca 5 ; 1 Gastroenterology, 

Marmara University Medical School, Istanbul, Turkey; 

2 Gastroenterology, Acibadem University Medical School, Istanbul, 

Turkey; 3 Gastroenterology, Istanbul University Medical Faculty, 

Istanbul, Turkey; 4 Gastroenterology, Ankara Universit Faculty of 

Medicine, Istanbul, Turkey; 5 Gastroenterology, Ege University 

Medical Faculty, Istanbul, Turkey 

Aim&Methods: Hepatitis C infection is more prevalent in the 

“Baby Boomer Population” those born between 1945 and 

1965 in USA and Western Countries. Although Turkey was 

not involved in World War II, birth rates were increased after 

1945 in Turkey. However, this population can not be defined 

as “Baby Boomers” since they have different properties. We 

aimed to determine the anti-HCV seropositivity in the population 

born between the years of 1945-1965 from the data 

of Turkey Nationwide Hepatitis Prevalence Study (TURKHEP) 

which was presented in AASLD in 2010 by Turkish Association 

for the Study of Liver Disease (TASL). A total of 5533 volunteers 

were screened in TURHEP Study, performed via home 

visits in 2009-2010 with volunteers living in urban and rural 

areas of 23 cities located in EUROSTAT NUTS 2 regions of 

Turkey which were determined based on two stage stratified 

sampling method. The participants were selected according 

to randomized sampling method of Statistics Institute of Turkey 

(TUIK). Participants’ socio-demographic features, anthropometrics, 

medical history, concomitant medications and presence 

of risk factors were recorded and blood samples were 

collected for the analysis of serum HBV, HDV, HCV markers, 

ALT and AST levels. Anti-HCV seropositivity and HCV RNA 

status was recruited from the TURKHEP data and compared 

with the population born after 1965. Results: TURKHEP data 

analysis included 5460 participants with data available on 

seroprevalence and risk factors for hepatitis C infection inwhich 

1980 participants (1037 female/943 male) borned between 

the years of 1945-1965 were recruited. Overall, 0.95% of the 

subjects were reactive for anti-HCV antibodies in whole population 

whereas Anti-HCV seropositive subjects were 28 out of 

1980 subjects in the population born between 1945-1965 at 

a rate of 1.4 % which is 1.5 times higher than the whole population. 

Half of these subjects were HCV RNA positive by the 

Real time PCR instrument, Rotor-Gene Q (QIAGEN, Germany). 

However among the subjects borned after 1965, Anti-HCV 

seropositive subjects were 23 out of 3460 subjects at a rate of 

0.7%. Conclusion: Eventhough the HCV seropositivity rate was 

higher than the whole Turkish population in Turkish population 

born between the years of 1945-1965, this rate of increase did 

not reach the higher rates of Western Baby Boomer Population 

showing the different transmission routes of HCV infection in 

Turkey. 

Disclosures: 


AbbVie 

The following authors have nothing to disclose: Osman C. Ozdogan, Nurdan 

Tozun, Sabahattin Kaymakoglu, Ramazan Idilman, Zeki Karasu


1823 

Age and Risk Factor-based Serologic Screening for Hepatitis 

C Virus among an Urban High-risk Population 

Rositsa B. Dimova 2,3 , Anthony D. Martinez 2,1 , Ethan Weinberg 4 , 

Ann Drobnik 5 , Andrew H. Talal 2,1 ; 1 Division of Gastroenterology, 

Dept of Medicine, SUNY-Buffalo, Buffalo, NY; 2 Center for Clinical 

Care and Research in Liver Disease, University at Buffalo, Buffalo, 

NY; 3 Department of Biostatistics, University at Buffalo, Buffalo, 

NY; 4 Department of Medicine, Weill Cornell Medical College, 

New York, NY; 5 New York City Department of Health and Mental 

Hygeine, New York, NY 

Background and aims: Hepatitis C virus (HCV) screening 

among individuals born between 1945 and 1965 (i.e. birth 

cohort) has been proposed to augment risk factor-based screening. 

Among an urban, high-risk population, we compared the 

frequency of HCV seropositivity among injection drug users 

(IDUs) and birth cohort members. Methods: We assessed HCV 

seropositivity in 7722 subjects from New York City drawn from 

HIV prevention programs, syringe exchange programs, drug 

treatment programs, and community health centers. HCV serology 

was assessed by immunoblot and data analysis employed 

logistic regression. Results: Mean age was 40.7 years, standard 

deviation (SD) 10.9 years, 47.4% were born between 

1945-1965, 81.2% were born in the US, 70.3% were male, 

53% were African American, 21.5% were Caucasian, 36.8% 

reported history of injection drug use, and 9.8% were HIV 

infected. Among men, 12.4% were men who had sex with 

men (MSM). Most participants were from HIV (72.7%) and substance 

use (56.4%) treatment sites. Overall, 26.6% were HCV 

seropositive, 55.8% of whom were born between 1945 and 

1965, and 82.2% had ever injected drugs. Among all participants, 

HCV seropositivity was significantly higher among IDUs 

than non-IDUs (60.5% versus 7.7%, odds ratio (OR) = 18.5, 

95% confidence interval (CI) [16.2, 21.1], p


1825 

Hepatitis C Virus (HCV) Eradication in Patients with 

Varying Severity of Cirrhosis: Impact on Portal Hypertensive 

Complications, Liver Transplant (LT) and Death 

Varun Saxena 1 , Lisa M. Nyberg 2 , Aditi Dasgupta 1 , Stephanie Straley 

1 , Lisa Catalli 1 , Anders H. Nyberg 2 , Norah Terrault 1 ; 1 Gastroenterology, 

University of California San Francisco, San Francisco, 

CA; 2 Kaiser Permanente Southern California, San Diego, CA 

Background: Recent all-oral antiviral therapy for HCV results in 

high rates of sustained virologic response (SVR) among patients 

with varying severity of cirrhosis. The impact of SVR on longer-term 

hepatic outcomes (beyond SVR12) in patients with 

cirrhosis, especially those who are Child-Pugh (CP) B/C, is 

currently unknown. Aims: We aimed to compare the liver-related 

events in a cohort of HCV-infected patients with varying 

severity of cirrhosis who achieved SVR with all oral antiviral 

therapy vs. those not achieving SVR / untreated matched controls. 

Methods: Adults with HCV genotype 1 cirrhosis treated 

with simeprevir (SMV) + sofosbuvir (SOF) ± ribavirin (RBV) 

were followed for a median of 0.80 yrs (IQR: 0.65-0.93) after 

treatment discontinuation. Randomly selected, untreated controls 

(1-3/pt) matched on site, age ± 5yrs, CP (A vs. B/C) and 

model for end-stage liver disease (MELD) score ± 2 were followed 

for a similar duration. Safety outcomes included change 

in MELD and CP scores, number of emergency room (ER) visits 

and hospitalizations, presence of portal hypertensive complications 

(new/worsening ascites, varices, encephalopathy and/or 

spontaneous bacterial peritonitis), need for LT and death. Survival 

from treatment discontinuation was compared in treated 

patients and matched controls over equivalent timeframe. 

Results: Of 371 patients, 93 were treated with SMV+SOF±RBV 

and 278 were matched, untreated controls. Cohort median 

age was 62yrs (IQR: 58-68), 32% female, 25% Hispanic, 5% 

Black, 24% diabetes, median (range) platelet count 133K/ 

mm 3 (20-341), albumin 3.5g/dL (2.3-4.7), MELD 8 (6-18), 

35% CP-B/C, 22% ascites, 22% encephalopathy, 8% varices. 

Of the SMV+SOF±RBV treated patients, 78 (84%) achieved 

SVR12. Liver-related outcomes between groups are shown in 

the Table. In bivariate Cox survival analysis, achievement of 

SVR vs. non-SVR/untreated (HR 0.15, 95% CI 0.02-0.97) and 

CP-A vs. CP-B/C (HR: 0.57, 95% CI 0.28-0.96) protected from 

LT/death. Conclusions: Achieving HCV cure among patients 

with compensated and decompensated cirrhosis significantly 

reduced the frequency of liver-related outcomes including portal 

hypertensive complications and LT/death. 

Liver-Related Outcomes in Patients Achieving SVR vs. Non-SVR/ 

Untreated Matched Controls 

*unadjusted log-rank 

Disclosures: 

Lisa M. Nyberg - Grant/Research Support: Merck, Gilead, Abbvie 

Lisa Catalli - Advisory Committees or Review Panels: Gilead, Kadmon 




The following authors have nothing to disclose: Varun Saxena, Aditi Dasgupta, 

Stephanie Straley, Anders H. Nyberg 

1826 

Can HCV Antigen Testing Replace HCV RNA PCR Analysis, 

for Diagnosis and Monitoring of Treatment for 

Hepatitis C? 

Andrew M. Hill 1 , Teri Roberts 2 , Valerianna Amorosa 3 , Kamron 

Pourmand 3 , Gail Matthews 4 , Graham Cooke 5 , Harun Khan 5 , 

Janice Main 5 , Ashley Brown 5 , Joy A. Peter 9 , David R. Nelson 6 , 

Michael W. Fried 7 , Jennifer Cohn 8 , Camilla S. Graham 10 ; 1 Pharmacology 

and Therapeutics, Liverpool University, Liverpool, United 

Kingdom; 2 Treatment Access Campaign, Medecins Sans Frontieres, 

Geneva, Switzerland; 3 Philadelphia VAMC, University of 

Pennsylvania, PA; 4 University of New South Wales, Kirby Institute, 

Sydney, NSW, Australia; 5 Faculty of Medicine, Imperial College, 

London, United Kingdom; 6 Clinical and Translational Science Institute, 

University of Florida, Gainesville, FL; 7 Liver Center, University 

of North Carolina, Chapel Hill, NC; 8 Division of Infectious Diseases, 

University of Pennsylvania, Philadelphia, PA; 9 Hepatology 

Research, University of Florida, Gainsville, FL; 10 Beth Israel Deaconess 

Medical Centre, Boston, MA 

Background: Simplified systems of diagnostic testing for hepatitis 

C virus (HCV) could make widespread implementation of 

Direct Acting Antiviral (DAA) based treatment more feasible in 

low and middle income countries. HCV core antigen tests (e.g. 

Abbott Architect assay) are cheaper and simpler to perform, 

but have lower limits of detection in the range of 1000 IU/mL. 

More expensive, complex HCV RNA nucleic acid assays (e.g. 

Roche TAQMAN) can detect HCV virus at levels of 15-25 IU/ 

mL. Methods: A systematic review identified studies testing samples 

in duplicate for HCV antigen and HCV RNA. Clinical data 

from 4 cohort studies evaluating patients relapsing on PEG- 

IFN or DAA based treatment for acute and chronic HCV were 

reviewed to determine the percentage of relapsing patients 

with HCV RNA levels above 1000 IU/mL before treatment and 

at End of Treatment (EOT)+12-48 week time points. Results: In 

24 studies evaluating 8142 samples tested in parallel by the 

Abbott Architect antigen and HCV RNA assays, 46 samples 

(0.6%) were HCV antigen positive but HCV RNA undetectable. 

There were 280 samples (3.5%) HCV antigen negative but 

HCV RNA detectable: these samples generally showed HCV 

RNA results 1000 IU/mL in 31 (100%) before treatment, 28 (87%) at 

EOT+12, 29 (94%) at EOT+24, and 31 (100%) at EOT+48. 

In a Philadelphia chronic HCV treatment cohort, among 90 

relapsing patients HCV RNA levels were >1000 IU/mL in 

90 (100%) before treatment, and 89 (99%) at EOT+. In an 

Australian acute/early chronic HCV treatment cohort, among 

13 relapsing patients, HCV RNA levels were >1000 IU/ml in 

12 (92%) before treatment, and 9 (69%) at EOT+12 (n=11) 

or EOT+24 (n=2). In the HIV-TARGET observational cphprt 

study, all 232 relapsing patients had HCV RNA levels >1000 

IU/mL before treatment. There were 92 relapser patients with 

HCV RNA data available at least 10 weeks after EOT: 89/92 

(97%) had HCV RNA >1000 IU/mL at first relapse. Summary: 

In the 4 cohort studies, 365/366 patients (99.7%) had HCV 

RNA levels >1000 IU/mL before treatment; 214/224 relapsing 

patients (96%) had HCV RNA levels above 1000 IU/mL 

12 weeks after EOT, and so could have been detected by the 

Abbott Architect assay. A minority of patients relapsed with 

HCV RNA>1000 IU/mL 24-48 weeks after EOT. Antigen testing 

before and after treatment could potentially replace HCV 

RNA PCR analysis, but further validation studies are required in 

acute and chronic infection. New, low-cost Point-of-Care HCV 

antigen assays are needed for use in low income countries, 

with lower detection limits in the range of 1000 IU/mL.


Disclosures: 

Andrew M. Hill - Consulting: Janssen 







The following authors have nothing to disclose: Teri Roberts, Valerianna Amorosa, 

Kamron Pourmand, Gail Matthews, Graham Cooke, Harun Khan, Janice 

Main, Ashley Brown, Joy A. Peter, Jennifer Cohn, Camilla S. Graham 

FRY graph 

1827 

Comparative analysis of online patient education 

resources relating to Hepatitis C 

Rishabh Gulati, Mohammad Nawaz, Sowjanya Kanna, Nikolaos 

Pyrsopoulos; Medicine, Rutgers New Jersey Medical School, Newark, 

NJ 

Introduction: Role of the Internet is ever-increasing in the present 

era as the first source of medical information. Imprecise, 

partial comprehension of textual information limits its efficacy 

in communicating the disease process to the patient. Here, we 

report a comparative analysis of readability of patient-centered 

text pertaining to Hepatitis C available online. Methods: In 

March 2015, patient-centered information from websites of 

American College of Gastroenterology (ACG), Centers for Disease 

Control & Prevention (CDC), American Liver Foundation 

(ALF), Mayo Clinic, National Institutes of Health (NIH), Uptodate, 

HCVAdvocate & WebMD was downloaded & processed 

in Microsoft Word. All data were formatted & categorized 

into subsections. Proper nouns, copyright information & certain 

medical terms were omitted to limit bias. Text was then analyzed 

for their specific level of readability using 7 quantitative 

scales: Flesch Reading Ease, Flesch–Kincaid level, Gunning fog 

index, SMOG, Coleman-Liau, FRY & New Dale–Chall using 

Readability Studio software. Results: Modified documents had 

a mean grade level that was 1 less than their original counterparts. 

ACG had the highest mean grade level of readability 

of it’s content (13.4±0.61), with the lowest being for WebMD 

(8.4±0.40). When compared with all subsets, the treatment 

subsection had the highest mean grade level (12.2±0.85). 

ANOVA analysis showed that there were significant differences 

in the grade level depending on the source website (p < 

0.05), however there was no significant impact by subsection 

when compared with all readability tests. Post hoc Turkey HSD 

Analysis showed ACG was written at a significantly higher 

grade level than other websites. The treatment section was usually 

the most difficult section written when compared with other 

subsections (p < 0.05). Conclusion: Patient material is above 

the recommended 6 th grade level across all websites. Treatment 

section is often the most difficult section to comprehend. 

Greater emphasis on clear & simple language is warranted to 

increase quality & comprehension of online patient education 

resources. 

Disclosures: 

Nikolaos Pyrsopoulos - Advisory Committees or Review Panels: GILEAD, BMS, 

ABBVIE, VITAL THERAPIES, Quest; Grant/Research Support: ABBVIE 

The following authors have nothing to disclose: Rishabh Gulati, Mohammad 

Nawaz, Sowjanya Kanna 

1828 

Benefits of Antiviral Treatment and Predictors of Subsequent 

Hepatocellular Carcinoma Risk in Chronic Hepatitis 

C Patients with or without Sustained Virological 

Response to Peg-interferon plus Ribavirin Therapy 

Mei-Hsuan Lee 1 , Sheng-Nan Lu 2 , Ming-Lung Yu 3 , Cheng-Yuan 

Peng 4 , I-Shyan Sheen 5 , Chen-Hua Liu 6 , Jia-Horng Kao 6 , Wan-Long 

Chuang 3 , Hwai-I Yang 7 , Gilbert L’Italien 8 , Yong Yuan 9 , Chien-Jen 

Chen 7 ; 1 National Yang-Ming University, Institute of Clinical Medicine, 

Taipei, Taiwan; 2 Division of Hepatogastroenterology, Department 

of Internal Medicine, Kaohsiung Chang-Gung Memorial 

Hospital and Chang Gung University College of Medicine, Kaohsiung, 

Taiwan; 3 Hepatobiliary Division, Department of Internal Medicine 

and Hepatitis Center, Kaohsiung Medical University Hospital 

and Kaohsiung Medical University college of Medicine, Kaohsiung, 

Taiwan; 4 Division of Hepatogastroenterology, Department of 

Internal Medicine, China Medical University Hospital, Taichung, 

Taiwan; 5 Division of Hepatology, Department of Gastroenterology 

and Hepatology, Linkou Medical Center, Chang Gung Memorial 

Hospital, Tao-Yuan, Taiwan; 6 Graduate Institute of Clinical Medicine, 

Department of Internal Medicine and Hepatitis Research Center, 

National Taiwan University College of Medicine and Hospital, 

Taipei, Taiwan; 7 Genomics Research Center, Academia Sinica, 

Taipei, Taiwan; 8 Yale University School of Medicine, New Haven, 

CT; 9 Global Health Economics and Outcome Research, Bristol-Myers 


Background & Aims: The aims of this large follow-up study 

were to evaluate the antiviral treatment efficacy in chronic hepatitis 

C patients by two distinctive cohorts, and to investigate 

the predictability of post-treatment seromarkers for HCC risk 

stratified by patients with sustained virological response (SVR) 

and non-SVR to antiviral treatment. Methods: The treatment 

cohort included 3,133 chronic hepatitis C patients treated with 

peg-interferon plus ribavirin (PR) for 6-12 months. In addition, 

the untreated cohort consisted of 1,366 anti-HCV seropositives 

who participated in a community-based liver cancer screening. 

All of them were seronegative for HBsAg and free of HCC 

at study entry. The treated patients were followed from the 

date starting PR therapy whereas the untreated patients were 

followed from the date of enrollment. Both cohorts were followed 

to the date of HCC diagnosis, death, or the end of 

2012, whichever came first. The Cox’s proportional hazards 

model was utilized to estimate the adjusted hazard ratio (HR) 

and 95% confidence interval (CI) associated with HCC by 

comparing patients with or without antiviral treatment. Among


the treated patients, the post-treatment seromarkers were evaluated 

for HCC risk by patients with or without SVR. Results: 

The patients in the treatment cohort had increased likelihood 

of advanced age, male gender, HCV genotype 1 infection, 

elevated serum levels of HCV RNA, AST, ALT, platelet count, 

and alfa-fetoprotein (AFP) at baseline (p


ratio for models including behavioural risk heterogeneity. Conclusions: 

Preferential mixing by HIV status and behavioural risk 

heterogeneity could be the main factors that have resulted in 

much higher HCV prevalence amongst HIV-positive MSM than 

in HIV-negative MSM. Greater HCV transmission could occur 

amongst HIV-negative MSM if HIV-negative MSM start using 

condoms less or mix more with HIV-positive MSM, something 

that may occur with widespread use of HIV pre-exposure prophylaxis. 

The effects of new HIV prevention interventions on 

HCV transmission should be considered. 

Disclosures: 

The following authors have nothing to disclose: Louis W. MacGregor, Peter Vickerman, 

Natasha K. Martin, Ford Hickson, Peter Weatherburn 

1831 

Positive Impact of Point of Care Testing and an Informational 

Brochure on Screening of the HCV Birth Cohort in 

a Rural New England Tertiary Care Center 

Arvind Suguness 1 , Casey M. Kolb Nava 1 , Kristen Ray 2 , Rolland C. 

Dickson 2 ; 1 Internal Medicine, Dartmouth Hitchcock Medical Center, 

White River Junction, VT; 2 Gastroenterology and Hepatology, 

Dartmouth Hitchcock Medical Center, Lebanon, NH 

Background. Hepatitis C Virus (HCV) screening of the Birth 

Cohort (BC) has been recommended by the CDC and USPSTF 

due to the high prevalence of HCV and advanced liver disease 

in this population. The use of risk-based screening was 

previously shown to be ineffective at identifying patients with 

HCV infection. To address these guidelines, we implemented 

a point-of-care testing (POCT) strategy for HCV, and surveyed 

patients within the cohort. The aim of this study was to assess 

factors in a HCV testing algorithm that influenced screening. 

Methods. A HCV BC screening initiative was fully implemented 

in an Internal Medicine Residents Clinic in a rural tertiary care 

center in the NE United States on 4-1-2014. BC patients (1945- 

1965) were identified by a prompt in the electronic medical 

record prior to arrival to clinic. They were given a brochure on 

HCV natural history, POCT, and HCV treatment at the time of 

check in. When patients were roomed they were offered testing 

with the Orasure finger prick test. The patients were given a 

questionnaire while they waited for the provider. HCV screening 

(+ or -) results were available within 20 minutes during the 

provider visit. Patients were informed of the results and those 

with positive results were offered same day PCR testing, with 

genotype performed if viral load was present. Results. There 

were 325 total respondents to the survey. Sixty three (19.4%) 

reported being asked by a health care provider to be tested 

for HCV in the past. Twenty one reported prior HCV testing 

(6.5%) and 8 reported a prior positive test (2.5%). Of the total 

respondents, 86 (26%) opted out of screening entirely, while 

239 patients (73.5%) chose to be screened at that visit. Of 

those screened, 117 (49.0%) said the information in the brochure 

or having test results immediately available influenced 

their decision to be tested. At least one reported risk factor 

was present in 142 patients (43.69%). Patients with at least 

one risk factor were no more likely to desire screening than 

those without risk factors (73.2% vs 73.8%, p=0.914). Patients 

with risk factors were no more likely than those without risk 

factors to report that the brochure or the immediately available 

results influenced their decision to get tested (43.2% vs 53.3%, 

p=0.123). Summary and Conclusion. Prior to initiation of a 

POCT strategy, relatively few patients in the birth cohort at 

our institution had been approached by health care providers 

about HCV screening and fewer still had been tested. The presence 

of an informational brochure, combined with point-of-care 

testing, significantly increased the number of patients with and 

without risk factors to undergo testing. 

Disclosures: 

Casey M. Kolb Nava - Consulting: AbbVie 

Kristen Ray - Advisory Committees or Review Panels: Abbvie 

Rolland C. Dickson - Advisory Committees or Review Panels: Biotest; Speaking 

and Teaching: gilead 

The following authors have nothing to disclose: Arvind Suguness 

1832 

Stability and prevalence of the NS3 Q80K polymorphism 

over time within HCV genotype 1a infected 

patients in Canada 

Jeffrey B. Joy 1 , Weiyan B. Dong 1 , Celia B. Chui 1 , Chanson J. 

Brumme 1 , Art Poon 1,3 , Huong Hew 2 , Richard Harrigan 1,3 ; 1 BC 

Centre for Excellence in HIV/AIDS, Vancouver, BC, Canada; 

2 Clinical Affairs, Janssen Incorporated, North York, ON, Canada; 

3 Medicine, University of British Columbia, Vancouver, BC, Canada 

Background: HCV genotype 1a (GT1a) infections harbouring a 

baseline Q80K polymorphism in the NS3 gene display reduced 

virologic response to IFN-based HCV treatments containing 

simeprevir. The prevalence of this and other NS3 resistance 

mutations in Canadian populations is not well described. Further, 

in the context of individual infections, the stability of this 

polymorphism among untreated patients over time is unknown. 

Methods: Using plasma samples serially collected over a 

10-year period from 121 HCV treatment naïve GT1a infected 

seroconverting injection drug users, we sought to investigate 

gain or loss of the Q80K polymorphism over time. A 1200- 

or 564-bp HCV NS3 fragment was sequenced by Sanger 

methods. Each sample was HLA typed to confirm database 

annotations on source individuals. HCV sequences were multiply 

aligned using MAFFT v7.154b. Phylogenetic trees were 

inferred using an approximate maximum likelihood method 

(FastTree2) and rooted under a molecular clock model using 

Path-O-Gen. To establish the prevalence of Q80K in Canadian 

Provinces, samples from GT1 LiPA HCV+ individuals were 

examined. Sequences were also screened for mutations associated 

with boceprevir/telaprevir resistance. Results: No patients 

whose first and last samples formed a monophyletic group 

altered their Q80K status. Nine patients changed genotypes 

(6 GT3a to GT1a, 2 GT1a to GT3a, and 1 GT1b to GT1a). 

Furthermore, in 10 patients, GT1a infections did not form a 

monophyletic group. Both between genotype and within genotype 

changes in viral lineage between collection dates suggest 

either (1) clearance followed by reinfection with a new variant 

or (2) a mixed infection. In sum, we observed 9 changes in 

Q80K in 121 patients, but in every case this resulted from 

patients switching HCV lineages rather than a mutation in 

their original HCV lineage. Prevalence of Q80K in Canada 

was 956/1835 (52%) in GT1a and provinces displayed no 

significant difference in prevalence of Q80K. Mutations associated 

with boceprevir or telaprevir resistance in this newly 

diagnosed population were also observed. Conclusions: These 

results suggest that, in the absence of therapy, Q80K is highly 

stable within HCV lineages and does not evolve in response 

to immune or other host specific effects. Observed changes 

in infection status amongst these patients supports genotypic 

and resistance testing of patients prior to starting IFN-based 

therapy, particularly amongst those at high risk of exposure to 

new variants of HCV such as intravenous drug users. 

Disclosures: 

Huong Hew - Employment: Janssen 

Richard Harrigan - Consulting: ViiV Health Care, Tobira Therapeutics, Selah 

Genomics Inc, Quest Diagnostics; Stock Shareholder: Merck, Gilead


The following authors have nothing to disclose: Jeffrey B. Joy, Weiyan B. Dong, 

Celia B. Chui, Chanson J. Brumme, Art Poon 

1833 

Performance of HCV Ag quantification as a screening 

tool in HCV mono-infected, HBV-HCV and HIV-HCV 

co-infected patients from Cameroon: the ANRS 12336 

study 

Léa Duchesne 1 , Richard Njouom 2 , Frédéric S. Lissock 2 , Ghislaine 

Flore Tamko-Mella 2 , Sandrine Rallier 3 , Lila Poiteau 3 , Alexandre 

Soulier 3 , Stéphane Chevaliez 3 , Guy Vernet 2 , Nicolas Rouveau 4 , 

Pierre-Marie Girard 1 , Karine Lacombe 1 ; 1 Inserm UMR-S1136, 

Paris, France; 2 Pasteur Center of Cameroon, Yaounde, Cameroon; 

3 National Reference Center for Viral Hepatitis B, C and Delta, 

Department of Virology, Hôpital Henri Mondor, Créteil, France; 

4 National Agency of Research on AIDS and viral hepatitis, Paris, 

France 

Background: HCV chronic infection diagnosis currently relies 

on anti-HCV antibody (HCV-Ab) detection. As it cannot differentiate 

an active infection from a resolved one, its diagnosis 

must be confirmed by HCV-RNA measurement which 

is scarcely available in resource-limited settings. Quantifying 

HCV core antigen (HCV-cAg), a marker of viral replication, 

could shorten this algorithm if used as a one-step tool. Aim: To 

assess the performance of the HCV-cAg quantification for the 

diagnostic of chronic HCV in a serum bank of HCV mono- and 

HCV-HBV or HCV-HIV co-infected patients from Cameroon and 

the influence of co-infections on the test’s results. Methods: The 

quantification of the HCV-cAg was performed by an automated 

assay (Abbott Diagnostics) in 476 HCV-Ab negative samples 

and 548 HCV-Ab and HCV-ARN positive samples (n=1024) 

collected in patients from the Pasteur Center of Cameroon. Its 

performance was assessed by calculating its sensitivity and 

specificity, and building ROC curves in order to compare its 

results to the gold standard (ELISA and/or PCR) with the Area 

Under the Curve (AUC). Results: Among these 1024 sera, 493 

were HCV mono-infected, 27 of 40 HIV-infected were co-infected 

with HCV and 28 of 47 HBV –infected were HCV-HBV. 

No statistical association was found between the HCV-cAg 

level and our covariates (age, gender, HBV or HIV co-infection). 

The correlation between HCV-cAg and HCV ARN was 

good in the mono-infected group (r=0.75, p


care clinic to screen persons born between 1945-1965 for 

HCV antibodies, and linking of viremic individuals to care. 

Emory University and Morehouse School of Medicine residents 

practicing in the Grady Memorial Hospital Primary Care Center 

(Atlanta, GA, USA) received one-on-one training regarding 

birth cohort screening, as well as an electronic medical record 

reminder prompt. Residents screened primary care patients, 

and project staff followed up positive HCV Ab tests and performed 

outreach and linkage to care. HCV Ab positive persons 

who attended an appointment at the Grady Primary Care 

Center, Grady Liver Clinic, or Infectious Disease Clinic during 

which the positive HCV test was addressed and further work-up 

ordered were counted as linked to care. Results: In a birth 

cohort population that was 92.5% Black/African American, 

412 (7.9%) of 5,239 patients screened for hepatitis C had 

HCV antibodies. HCV RNA testing was completed for 90% 

of the seropositive patients, and 70% were viremic. Of 258 

patients with a positive RNA test, 229 (89%) were referred to 

care, and 212 (82%) referred patients attended a linkage visit. 

Conclusions: The TILT-C screening program was found to be 

feasible and effective. The program was successful in detecting 

previously undiagnosed chronic hepatitis C infections and linking 

persons to care in an urban primary care center. 

TILTC: HCV Testing and Linkage Cascade 

Disclosures: 

Lesley Miller - Advisory Committees or Review Panels: Bristol Myers Squibb 

Anne C. Spaulding - Grant/Research Support: Gilead Sciences, BMS 

The following authors have nothing to disclose: Brandi Park, Nyiramugisha Niyibizi, 

April D. Elam, Francois Rollin, Shelly-Ann Fluker 

1836 

WITHDRAWN 

1837 

Cannabinoid receptor 2 (CB2) 63 RR variant is associated 

with immune-mediated disorders in patients with 

chronic HCV infection 

Nicola Coppola 1 , Rosa Zampino 2 , Giulia Bellini 3 , Maria Stanzione 

4 , Nicolina Capoluongo 1 , Aldo Marrone 2 , Margherita 

Macera 1 , Adriana Boemio 2 , Sabatino Maione 3 , Luigi Adinolfi 2 , 

Emanuele Miraglia del Giudice 5 , Evangelista Sagnelli 1 , Francesca 

Rossi 5 ; 1 Mental Health and Public Medicine, Second University of 

Naples, Naples, Italy; 2 Internal Medicine and Hepatology, Second 

University of Naples, Naples, Italy; 3 Department of Experimental 

Medicine, Second University of Naples, Naples, Italy; 4 Department 

of Clinical and Experimental Medicine and Surgery, Second 

University of Naples, Naples, Italy; 5 Department of Woman, Child 

and of General and Specialized Surgery, Second University of 

Naples, Naples, Italy 

Background: Patients with HCV chronic infection frequently 

show immune-mediated disorders (IMDs). The Cannabinoid 

(CB) receptor 2, predominantly expressed in the immune cells, 

plays an important role on the function of the immune system. 

In particular, the CB2-63 variants (rs35761398) affects the 

ability of the CB2 receptor to exert its inhibitory function on T 

lymphocyte. Aims: to evaluate whether CB2 variants are associated 

with the presence of IMDs in patients with chronic HCV 

infection. Methods: Considering that nearly 30% of anti-HCV 

positive patients are affected by IMDs, we planned a 12-month 

recruitment period for treatment-naïve anti-HCV positive patients 

with signs of IMD and a 4-month period for treatment-naïve 

anti-HCV patients lacking these signs. The enrollment stared 

in September 2013 and at the end of the recruitment periods 

168 patients have been selected, 81 anti-HCV/HCV-RNA positive 

with signs of IMDs and 87 anti-HCV/HCV-RNA positive 

with no sign of IMDs. The presence of IMDs was defined by 

at least one of the following conditions: ANA positivity (titers 

≥1:160) observed in 22 (27.2%) cases, SMA positivity (titers 

≥1:160) in 3 (3.7%), a cryocrite >2% in 24 (29.6%), history 

or active autoimmune thyroiditis in 25 (30.9%), psoriasis in 4 

(4.9%), B-cells non-Hodgkin lymphoma in 2 (2.5%) and autoimmune 

hemolytic anemia in 1 (1.2%) case; no patient showed 

signs of lichen planus nor Syogren syndrome. All patients were 

screened for the CNR2 rs35761398 SNP by a TaqMan Assay 

Results: Compared with the 87 patients lacking IMDs, the 81 

in the IMDs group more frequently were females (65% vs 45%, 

p=0.01), but not other significant difference was found in initial 

demographic, epidemiologic, serological, biochemical and 

virological data. In particular, the age (mean+SD: 53±14.1 vs. 

52.9±13.4 years), ALT serum levels, HCV viral load and in distribution 

of HCV genotypes were similar in these two groups. 

Instead, the prevalence of the patients with the CB2-63 RR variant 

was significantly higher in patients in the IMD group than 

in those in the non-IMD group (49.4% vs 24.1%, p=0.001). A 

logistic regression analysis including the CB2-63 receptor (RR 

vs QR or QQ), age and sex, identified the CB2-63 RR as the 

only independent predictor of IMDs (p =0.005). Conclusions: 

the data suggest a significant previously unknown association 

between CB2-63 RR variant and IMDs in anti-HCV patients, an 

observation deserving further investigation on a larger series of 

patients to define its clinical value 

Disclosures: 

The following authors have nothing to disclose: Nicola Coppola, Rosa Zampino, 

Giulia Bellini, Maria Stanzione, Nicolina Capoluongo, Aldo Marrone, Margherita 

Macera, Adriana Boemio, Sabatino Maione, Luigi Adinolfi, Emanuele 

Miraglia del Giudice, Evangelista Sagnelli, Francesca Rossi 

1838 

Serum levels of Wisteria floribunda agglutinin–positive 

human Mac-2 binding protein are useful for evaluating 

early liver fibrosis in hepatitis C patients 

Kohei Oda 1 , Hirofumi Uto 1,2 , Seiichi Mawatari 1 , Rie Ibusuki 1 , Sho 

Ijuin 1 , Hiroka Onishi 1 , Haruka Sakae 1 , Kaori Muromachi 1 , Akihiko 

Oshige 1 , Kotaro Kumagai 1 , Tsutomu Tamai 1 , Akihiro Moriuchi 

3 , Akio Ido 1 ; 1 Digestive and Lifestyle Diseases, Department of 

Human and Environmental Sciences, Kagoshima University Graduate 

School of Medical and Dental Sciences, Kagoshima, Japan; 

2 Center for Digestive and Liver Disease, Miyazaki Medical Center 

Hospital, Miyazaki, Japan; 3 Department of HGF Tissue Repair and 

Regenerative Medicine, Kagoshima University Graduate School of 

Medical and Dental Sciences, Kagoshima, Japan 

Objective: Liver fibrosis is the most important risk factor for 

liver cancer, and the rate of liver carcinogenesis rises significantly 

in cases of mild to advanced liver fibrosis (stage F2 

or greater) in patients with hepatitis C. Therefore, it is very 

important to evaluate liver fibrosis precisely, but appropriate 

biomarkers for evaluating less-advanced liver fibrosis have


not been identified. Wisteria floribunda agglutinin–positive 

human Mac-2 binding protein (WFA(+)-M2BP) was reported 

to be a novel serum marker of liver fibrosis identified in glycoproteomic 

biomarker screening studies, and was effective for 

evaluating advanced liver fibrosis. However, its usefulness in 

less-advanced liver fibrosis has not been fully elucidated. In this 

study, we examined the utility of WFA(+)-M2BP as liver fibrosis 

marker in patients with less-advanced liver fibrosis. Methods: 

One hundred forty-three patients with biopsy-proven chronic 

hepatitis C were enrolled in this study. We examined the association 

between WFA(+)-M2BP and the results of blood biochemistry 

and liver histology examinations. Results: Fifty-nine 

patients were male, and the mean age of the entire group 

was 60 years. The mean platelet count was 16.6×10 4 /mL, 

serum ALT was 48 IU/L, AFP was 4.9 ng/dL, and liver fibrosis 

scores of F0–1/F2/F3/F4, respectively, were 80/42/14/7. 

WFA(+)-M2BP rose with liver fibrosis progression, was significantly 

correlated with liver fibrosis markers and scores, and 

was significantly associated with histopathology findings of 

liver fibrosis. For cases of less-advanced liver fibrosis (stage 

F2 or less), WFA(+)-M2BP was significantly elevated by a 

liver fibrosis extension case. Receiver operating characteristic 

(ROC) analysis revealed that the WFA(+)-M2BP cut-off value of 

1.94 [C.O.I.] discriminated between stages F2 and F0–1 (area 

under the ROC curve, 0.768) better than other serum markers 

such as platelet count (0.613), hyaluronic acid (0.661), type 

IV collagen (0.627), and procollagen III peptide (0.610), and 

better than liver fibrosis scores such as the FIB4 index (0.645) 

and APRI (0.660). In the multivariate analysis, high WFA(+)- 

M2BP was an independent factor associated with mild liver 

fibrosis (F2) (OR, 6.37; 95% CI, 1.85–22.00; P=0.003). Furthermore, 

in the 15 cases we could evaluate, WFA(+)-M2BP 

improved significantly following interferon treatment (from 

3.21 to 1.94;P=0.015). Conclusions: In chronic hepatitis C, 

WFA(+)-M2BP should prove useful in evaluating relatively early 

cases of liver fibrosis. In addition, WFA(+)-M2BP may serve as 

an indicator of improvement in liver fibrosis at an early stage. 

Disclosures: 

The following authors have nothing to disclose: Kohei Oda, Hirofumi Uto, Seiichi 

Mawatari, Rie Ibusuki, Sho Ijuin, Hiroka Onishi, Haruka Sakae, Kaori Muromachi, 

Akihiko Oshige, Kotaro Kumagai, Tsutomu Tamai, Akihiro Moriuchi, Akio 

Ido 

1839 

Factors Associated with Hepatitis C Virus Infection 

among Persons Ages 18 to 29 who Inject Drugs in Suburban 

and Rural Wisconsin 

David B. Rein 1 , Danielle Liffmann 1 , Jim Brunner 2 , Emily Wievel 2 , 

Gregory Scott 3 , Daniel Raymond 4 , Lisa Danelski 2 , Scott Stokes 2 , 

Joanne E. Brady 1 , Jon E. Zibbell 5 ; 1 NORC at the University of 

Chicago, Atlanta, GA; 2 AIDS Resource Center of Wisconsin, 

Green Bay, WI; 3 Sociology, DePaul University, Chicago, IL; 4 Harm 

Reduction Coalition, New York, NY; 5 Division of Viral Hepatitis, 

CDC, Atlanta, GA 

Background: Due to national increases in heroin and injection 

drug use, along with the recent HIV outbreak among HCV-infected 

persons who inject drugs (PWID) in Indiana, information 

on behavioral risk factors for HCV infection among young 

PWID in non-urban settings is needed. Methods: Between September 

2014 and May 2015, we collected survey data and 

hepatitis C antibody testing information from young adults ages 

18 to 29 who had injected drugs illicitly at least 1 time within 

the last year. Using a respondent driven snowball sampling 

design, the study collected HCV antibody status using rapid test 

kits donated by OraSure® as well as self-reported information 

on demographics, injecting behaviors, social network size, 

personal characteristics, and drug using histories. This initial 

analysis estimated descriptive statistics and stepwise multivariate 

logistic regressions (using an alpha of 0.05 for selection) 

of the risk of HCV infection. For selection, we included demographics, 

number of lifetime injections, history of ever sharing 

needles, filters, mix water, or cottons, seeing other people’s 

blood while injecting, reported size of injecting network, a 

history of ever injecting heroin, prescription opioids, cocaine, 

and methamphetamines, and scores on abbreviated scales of 

self-efficacy, extroversion, addiction, sensation seeking, and 

socio-normative aspirations. Results: Through May, 2015, 

we enrolled 265 persons: 58.4% male, 81.9% white, 14.7% 

American Indian and 3.4% of other races, with a mean age 

of 23.4. Of those, 34.0% were HCV+, 56.2% reported more 

than 100 lifetime injections, and 66.4%, 55.9%, 59.3%, and 

46.4% reporting ever sharing needles, filters, mix water, and 

cookers respectively. In multivariate analyses, injecting more 

than 100 times (OR 4.9; 95% CI 2.7-9.2) and having ever 

injected cocaine (OR 2.7; 95% CI 1.4-4.9) were significantly 

related to HCV+. Among those with less than 100 lifetime injections, 

sharing mix water (OR 4.6; 95% CI 1.3-16.1), injecting 

heroin (OR 9.5; 95% CI 1.0-89.3), and injecting cocaine 

(OR 4.3; 95% CI 1.2-15.3) were significantly associated with 

HCV+. Among persons who reported more than 1,000 lifetime 

injections, sharing filters (OR 3.1; 95% CI 1.3-7.5) and 

injecting cocaine (OR 3.7, 95% CI 1.3-10.0) were significantly 

associated with HCV+. Conclusions: We identified an HCV+ 

prevalence 34.0% among rural/suburban PWID ages 18 to 

29. Injecting cocaine was significantly associated with HCV 

antibody positivity in the full population. Sharing injection 

equipment other than syringes was significantly associated with 

higher HCV+ risk when the sample was stratified by number of 

lifetime injections. 

Disclosures: 

David B. Rein - Grant/Research Support: Gilead Sciences, Inc. 

Daniel Raymond - Grant/Research Support: Gilead, Janssen 

The following authors have nothing to disclose: Danielle Liffmann, Jim Brunner, 

Emily Wievel, Gregory Scott, Lisa Danelski, Scott Stokes, Joanne E. Brady, Jon 

E. Zibbell 

1840 

Hyperbilirubinemia in HIV-HCV Co-infected Patients on 

cART - Drug Effect or Liver Disease Severity? 

Mathew Kaspar, Richard K. Sterling; Virginia Commonwealth University, 

Richmond, VA 

Background: Hyperbilirubinia (HB) is common and a known 

side effect of many medications used in the management of 

HIV, particularly the protease inhibitors (PI) Atazanavir and 

Indinavir. While typically benign, the sudden development of 

HB in HIV-HCV presents a unique challenge to determine if it 

is a benign drug effect, related to liver disease progression, or 

combination of the two. To date, there have been no studies 

investigating the impact of underlying liver histology to distinguish 

drug effect from progression of liver disease in the 

HIV-HCV population. Objective: To determine if HB in the HIV- 

HCV pts being treated with PI is primarily due to drug effect, 

progression of liver disease, or both. Methods: We performed 

a retrospective analysis of 344 HIV-HCV pts undergoing liver 

biopsy. Pts with HBsAg +, hepatic decompensation, or HCC 

were excluded. Demographic, clinical, laboratory, and biopsy 

data were collected. The primary endpoint was HB (defined as 

serum bilirubin greater than 1.2mg/dL, the ULN). Univariate 

and multivariate analyses was used to determine factors associated 

with HB. Advanced fibrosis (AF) was defined by F3 or 

F4 histology (Knodell). Results: The mean age was 46 years,


75% were male, 84% were black, 91% were GT 1, 19% had 

>5% steatosis, and 31% had AF. 155 pts (46%) were being 

treated with a PI: Atazanavir (n=40), Kaletra (n=40), Indinavir 

(n=15), Nelfinavir (n=34), Darunavir (n=6), and other (n=20). 

The prevalence of HB (range 1.3 to 9.4 mg/dL) was 15.4%. 

Those with HB had higher AST (132 vs 75; p


of increasing the number of PWIDs treated with new oral DAAs 

was considered, including the annual number needed to treat 

in order to reduce the HCV-infected PWID population by 2030. 

Results: If the current transmission paradigm continues, there 

are projected to be 3,620 HCV infected PWIDs in 2030. 

Annually treating 40 HCV-infected PWIDs with new oral DAAs 

(1% of HCV-infected PWID population in 2014) resulted in a 

5% reduction in HCV-infected PWIDs by 2030, while annual 

treatment of 200 PWIDs (5% of 2014 population) resulted in a 

reduction of over 25% by 2030. Treating 387 PWIDs annually 

(17% of 2014 population) resulted in a >90% reduction in 

HCV-infected PWIDs by 2030. Targeting treatment to PWIDs 

engaged in OST and NEP would provide the greatest reduction 

in prevalence for the number of individuals treated (2.2 treated 

in OST/NEP to reduce prevalence by 1, as compared with 6.8 

treated in the general population). Conclusions: The results 

show that treating yet a small amount of PWIDs resulted in 

substantial decreases in the HCV infected PWID population by 

2030. Furthermore, the relative impact of treatment was greatest 

when focused on the population engaged in OST and NEP. 

Treatment is expected to increase the rate of reinfection; however, 

reinfection will decline as HCV prevalence decreases. 

This analysis supports the implementation of a screening and 

treatment strategy among PWIDs when combined with an 

expansion of harm reduction programs. 

Disclosures: 

Stefan Bourgeois - Advisory Committees or Review Panels: AbbVIe, Gilead; Consulting: 

Roche, MSD; Speaking and Teaching: Janssen, BMS 

Sarah Blach - Employment: Center for Disease Analysis 


Geert Robaeys - Advisory Committees or Review Panels: Gilead; Speaking and 

Teaching: Merck, Janssens 

The following authors have nothing to disclose: Catharina Mathei, Christian 

Brixko, Jean-Pierre Mulkay, Thomas Sersté 

1843 

Non-Invasive Fibrosis Scores: Do They Predict Antiviral 

Treatment Response, Decompensation, Hepatocellular 

Carcinoma and Significant Liver Related Adverse Events 

in Chronic Hepatitis C? 

Ragesh B. Thandassery 1 , Madiha E. Soofi 1 , Anil John 1 , Samir S. 

Nairat 1 , Abdulrahman A. Alfadda 2 , Saad R. Al Kaabi 1 ; 1 Hamad 

Medical Corporation, Doha, Qatar; 2 King Faisal Specialist Hospital 

and Research Center, Riyadh, Saudi Arabia 

Background The role of non-invasive liver fibrosis scores (NIF) 

in predicting antiviral treatment (AVT) response and post treatment 

significant liver related events (SLRE) in chronic hepatitis 

C (CHC) is less studied. Aim To compare 12 simple NIF, 

derived from routine blood investigations, for predicting 

response to AVT and SLRE. Methods 1605 patients underwent 

liver biopsy (LB, Scheuer classification) and received AVT 

(pegylated interferon and ribavirin). 12 NIF [AST-platelet count 

ratio index (APRI), Fibrosis-4 (FIB-4) score, Lok score, GUCI 

score, Fibroalpha score, modified APRI, King score, AST-ALT 

ratio (AAR), Fibrosis Index (FI), Fibro score, Fibrosis cirrhosis 

index (FCI) and Globulin platelet index (GPI)] were calculated 

prior to AVT. AUROCs were calculated for each of these NIF 

for predicting non-response to AVT and development of SLRE 

(defined as development of any event requiring intervention; 

decompensation and hepatocellular carcinoma, HCC) on 

follow-up Results Mean age 41.9years, predominantly genotype 

4(65%).1089(67.8%) were responders, 482(30%) non 

responders and 34(2.1%) relapsers. After median follow-up of 

6580.5 patient-years; 52(3.2%) had decompensation (bleed- 

9, ascites-39, jaundice-22, hepatic encephalopathy-7, spontaneous 

bacterial peritonitis-10, hepatorenal syndrome-4), 

11(0.7%) had HCC and 60(3.7%) had SLRE. The predictive 

accuracy of NIF and LB for non-response to AVT was low. 

FibroQ, King score and FIB-4 showed high accuracy for predicting 

adverse events. For predicting decompensation, HCC 

and SLRE, FibroQ (0.88), King score (0.90) and FibroQ (0.87) 

had highest AUROC respectively. Conclusions Predictive accuracy 

of NIF for non-response to treatment was low. Some NIF 

have high accuracy for predicting development of decompensation, 

HCC and SLRE on follow-up. Application of these simple 

scores can improve assessment of liver prognosis in patients 

treated for CHC. 

Non-invasive scores in predicting post treatment events 

* AUROC(95% Confidence interval) 

Disclosures: 

The following authors have nothing to disclose: Ragesh B. Thandassery, Madiha 

E. Soofi, Anil John, Samir S. Nairat, Abdulrahman A. Alfadda, Saad R. Al Kaabi


1844 

Male gender, genotype 3, previous alcohol use, 

increased BMI, and diabetes are factors independently 

correlated to advanced HCV chronic liver disease in 

Italy: data from PITER (Piattaforma Italiana per lo studio 

della Terapia delle Epatiti viRali) cohort study 

Loreta A. Kondili 1 , Stefano Rosato 2 , Liliana E. Weimer 1 , Maria 

Giovanna Quaranta 1 , Loredana Falzano 1 , Alessandra Mallano 

1 , Maria Elena Tosti 2 , Maurizio Massella 1 , Maurizia R. Brunetto 

31 , Anna Linda Zignego 4 , Mario Rizzetto 29 , Alfredo Di Leo 24 , 

Giovanni Raimondo 3 , Carlo Ferrari 25 , Antonio Craxi 30 , Gloria 

Taliani 5 , Pierluigi Blanc 6 , Antonio Gasbarrini 7 , Luchino Chessa 8 , 

Elke M. Erne 9 , Giovanna Fattovich 10 , Pietro Andreone 11 , Maria 

Vinci 12 , Francesco P. Russo 14 , Erica Villa 13 , Giovanni B. Gaeta 15 , 

Teresa A. Santantonio 19 , Guglielmo Borgia 20 , Gabriella Verucchi 

21 , Carmine Coppola 22 , Marcello Persico 17 , Liliana Chemello 28 , 

Alfredo Alberti 14 , Vincenzo De Maria 26 , Massimo Puoti 27 , Raffaele 

Bruno 18 , Paolo Caraceni 11 , Massimo Andreoni 16 , Marco Marzioni 

23 , Stefano Vella 1 ; 1 Therapeutic Research and Medicines Evaluation, 

Istituto Superiore di Sanità, Rome, Italy; 2 Istituto Superiore 

di Sanità, Rome, Italy; 3 University of Messina, Messina, Italy; 4 University 

of Florence, Florence, Italy; 5 Sapienza University of Rome, 

Rome, Italy; 6 Santa Maria Annunziata Hospital, Florence, Italy; 

7 Policlinico Universitario Agostino Gemelli, Rome, Italy; 8 Azienda 

Ospedaliero-Univesitaria di Cagliari, Cagliari, Italy; 9 Azienda 

Ospedaliera di Padova, Padova, Italy; 10 Azienda Ospedaliero 

Universitaria Integrata di Verona, Verona, Italy; 11 Azienda 

Ospedaliero Universitaria di Bologna, Bologna, Italy; 12 Niguarda 

Hospital, Milano, Italy; 13 University of Modena, Modena, Italy; 

14 Azienda Ospedaliero Universitaria di Padova, Padova, Italy; 

15 Second University of Napoli, Napoli, Italy; 16 University Tor Vergata, 

Rome, Italy; 17 University of Salerno, Salerno, Italy; 18 University 

of Pavia, Pavia, Italy; 19 University of Foggia, Foggia, Italy; 

20 Federico II University of Naples, Naples, Italy; 21 Sant’Orsola 

Malpighi Hospital, Bologna, Italy; 22 Gragnano Hospital, Napoli, 

Italy; 23 Università Politecnica delle Marche, Ancona, Italy; 24 University 

of Bari, Bari, Italy; 25 Azienda Ospedaliero-Universitaria di 

Parma, Parma, Italy; 26 Azienda Ospedaliero-Universitaria Mater 

Domini, Catanzaro, Italy; 27 Azienda Ospedaliera Niguarda-Cà 

Granda, Milano, Italy; 28 University of Padua, Padova, Italy; 29 University 

of Torino, Torino, Italy; 30 University of Palermo, Palermo, 

Italy; 31 Azienda Ospedalero-Universitaria Pisana, Pisa, Italy 

Aim: Italy has the highest prevalence rates of HCV infection 

in Europe. In order to evaluate factors associated with severe 

liver disease in Italian patients in care, data derived from the 

PITER cohort study were analysed. Methods: The relationship 

between severe fibrosis stage/cirrhosis and sociodemographic 

characteristics, HCV RNA genotype, alcohol, body mass index 

(BMI), ALT, AST, GGT, platelets, diabetes, cardiovascular, 

neurological/psychiatric, autoimmune/reumatological and 

neoplastic diseases were evaluated by univariate and logistic 

regression statistical models. The regression model’s goodness 

of fit (calibration and sensitivity) was also estimated. Results: To 

date the cohort consists of 6831 consecutive HCV chronic liver 

disease patients (mean age 59±19 years; 3797 males) on clinical 

care. HCV genotype 1b (58%) and genotype 2 (15%) are 

significantly prevalent in older ages (older than 59 years) compared 

to genotypes 3 (10%) and 4 (7%), which are prevalent in 

younger ages. F4/cirrhosis stage was present in 2579 (38%) 

patients. It increased by age as expected, although 32% of 

patients younger than 60 years (3371 patients) had F4/cirrhosis. 

Of the enrolled patients, 48% were treatment-experienced. 

The independent role of each factor related with F4/cirrhosis, 

as defined by the logistic regression analysis, is as follows: 

male vs female (3797/3034) OR:1.42 (95% CI:1.25-01.66); 

BMI≥25 vs BMI


1845 

Effectiveness and cost-effectiveness of improvements in 

harm reduction interventions, a better cascade of care, 

and Treat as Prevention of chronic hepatitis C in people 

who inject drugs (PWID) in France (ANRS 95146) 

Anthony Cousien 1 , Viet Chi Tran 2 , Sylvie Deuffic-Burban 1,3 , Marie 

Jauffret-Roustide 4,5 , Guillaume Mabileau 1 , Jean-Stéphane Dhersin 

6 , Yazdan Yazdanpanah 1,7 ; 1 IAME, UMR 1137, INSERM - Univ 

Paris Diderot, Sorbonne Paris Cité, Paris, France; 2 Laboratoire Paul 

Painlevé UMR CNRS 8524, UFR de Mathématiques, Université des 

Sciences et Technologies Lille 1, Villeneuve d’Ascq, France; 3 LIR- 

IC-UMR995, INSERM - Université de Lille, Lille, France; 4 INSERM 

U988/UMR CNRS8211/Université Paris Descartes, Ecole des 

Hautes Etudes en Sciences Sociales, CERMES3: Centre de Recherche 

Médecine, Sciences, Santé, Santé Mentale et Société, Paris, 

France; 5 Institut de Veille Sanitaire, Saint-Maurice, France; 6 LAGA, 

CNRS, UMR 7539, Université Paris 13, Sorbonne Paris Cité, Villetaneuse, 

France; 7 Hôpital Bichat Claude Bernard, Service des 

Maladies Infectieuses et Tropicales, Paris, France 

We estimated the cost-effectiveness of strategies designed to 

improve harm reduction interventions and/or the cascade of 

care in PWID in the context of the incoming direct-acting antivirals 

(DAAs) in France. We used a dynamic model to simulate 

life expectancy in discounted quality adjusted life years 

(QALYs), direct lifetime discounted costs, incremental cost-effectiveness 

ratio (ICER) and the number of first generation new 

HCV infections for each strategy among PWID in Paris metropolitan 

area from 2015 until death: S1: base case=current 

practice. Time before access to needle and syringe programs 

(NSP) after injection initiation=1y, time before access to opioid 

substitution therapies (OST) when in NSP=0.5y, time to 

diagnosis after infection=1.25/1.45y, time to linkage to care 

after diagnosis=2.6y, loss to follow-up (LTFU) rate=14%/y, 

SVR rate=95%, treatment initiation: fibrosis ≥F2 S2: improved 

risk reduction interventions. Access to NSP after injection initiation=0.25y, 

time before access to OST when in NSP=0.25y 

S3: treatment initiation: fibrosis ≥F0 S4: improved cascade of 

care. Time to diagnosis=0.5y, time to linkage to care=0.5y, 

LTFU rate=5%/y, SVR rate=95% S5: S3&S4 S6: S2&S3&S4 

Results are presented in Table. Compared with the base case, 

improved cascade of care (S4) increased the average life 

expectancy with an ICER of 8,373€/QALY gained (


(HCV-2). Phylogenetic analysis revealed that all HCV-2 isolates 

from Suriname were part of 6 robust phylogenetic clusters containing 

HCV-2 sequences obtained from Surinamese living in 

The Netherlands. Conclusion HCV prevalence among the Surinamese 

population attending the ED was 1% and was highest 

among those of Javanese ethnicity. Older patients were more 

often infected, suggesting a long-standing infection acquired 

in the past. HCV-2 is the circulating genotype in Suriname and 

was likely introduced in Suriname by slave trade from western 

Africa in the past centuries. Targeted screening with linkage to 

care and prevention guidelines for identified at-risk groups are 

required, as well as implementation of stricter hygiene regulations 

for permanent make-up salons. Peginterferon and ribavirin 

are not yet routinely given as HCV treatment in Suriname, 

however given the predominance of favorable HCV genotype 

2, would be strongly advised. 

Disclosures: 

Richard Molenkamp - Independent Contractor: Roche Diagnostics 

Maria Prins - Speaking and Teaching: msd, roche 

The following authors have nothing to disclose: Sigrid MacDonald - Ottevanger, 

Stephen G. Vreden, Jannie van der Helm, Thijs J. van de Laar, Els Dams, Jimmy 

Roosblad, John F. Codrington 

1847 

Hepatocellular carcinoma (HCC) scoring system for 

the individualized prediction of liver cancer in 1080 

HCV-related compensated cirrhosis included in the 

French multicenter prospective cohort ANRS CO12 Cir- 

Vir. 

Nathalie Ganne 1 , Valerie Bourcier 1 , Richard Layese 2 , Nabila Talmat 

1 , Ventzislava Petrov-Sanchez 3 , Patrick Marcellin 4 , Dominique 

Guyader 5 , Stanislas Pol 6 , Dominique G. Larrey 7 , Victor de Ledinghen 

8 , Denis Ouzan 9 , Fabien Zoulim 10 , Pierre Nahon 1 , Françoise 

Roudot-Thoraval 2 ; 1 Hepatology, Jean Verdier hospital, Bondy, 

France; 2 Henri Mondor hospital, Creteil, France; 3 ANRS, Paris, 

France; 4 Beaujon hospital, Clichy, France; 5 CHU, Rennes, France; 

6 Cochin hospital, Paris, France; 7 CHU, Montpellier, France; 8 CHU, 

Bordeaux, France; 9 none, Saint Laurent du Var, France; 10 CHU, 

Lyon, France 

Background and aims: This study aimed to develop and validate 

a simple scoring system to refine individualized prediction 

of HCC risk in patients with HCV-related compensated 

cirrhosis included in the French prospective multicentre ANRS 

CO12 CirVir cohort (1) . Methods: Among 1323 patients with 

HCV-cirrhosis enrolled in the ANRS CO12 CirVir, 720 and 

360 patients were randomly assigned to the training and validation 

sets, respectively. Cox multivariate proportional hazards 

model was used to predict HCC occurrence in the population, 

then nomograms were computed to assess individualized risk. 

Results: During a mean follow-up of 51 months, 103 (14%) and 

39 (11%) patients developed HCC, respectively in the training 

and validation cohorts. According to Cox regression analysis, 

5 variables were independently associated with the occurrence 

of HCC in the training cohort (table 1). A 13-point risk score 

was derived in the training cohort using these 5 variables, the 

score of each variable being obtained by linear transformation 

of the coefficients and rounding. The population was stratified 

according to this scoring system into four groups, with HCC 

occurrence gradually increasing from 0% to 20% at 3 years, 

and 0% to 37% at 5 years, for patients with the lowest (score 

≤ 2) and highest (score ≥ 11) HCC risk, respectively and validated. 

The AUC for the 1- and 3-year prediction were 0.75 

[0.64; 0.85] and 0.75 [0.71;0.80] respectively. Derived from 

this score, a nomogram was also built, enabling individualized 

prediction of HCC occurrence at 1, 3 or 5 years. Conclusion: 

An HCC score constructed using 4 baseline variables 

(age, past excessive alcohol consumption, platelets count, GGT 

serum level) and qualitative HCV RNA at end-point is accurate 

to predict HCC at the individual level in French patients 

with HCV-cirrhosis, with or without SVR. External validation in 

non-French populations is warranted. (1) Trinchet et al. Complications 

and competing risks of death in compensated viral 

cirrhosis (ANRS CO12 CirVir prospective cohort). Hepatology. 

2015 Feb 11. doi: 10.1002/hep.27743. 

table 1 

Disclosures: 





Abbvie 

Dominique Guyader - Advisory Committees or Review Panels: ROCHE, GILEAD, 

IRIS, ABBVIE; Board Membership: MERCK; Grant/Research Support: JANSSEN; 




















Roche 

The following authors have nothing to disclose: Nathalie Ganne, Valerie 

Bourcier, Richard Layese, Nabila Talmat, Ventzislava Petrov-Sanchez, Denis 

Ouzan, Pierre Nahon 

1848 

Differential Progression to Cirrhosis and Hepatic Decompensation 

in Asian Americans (AA) and Non-Asian (NA) 

Patients with Chronic Hepatitis C (CHC) 

An K. Le 1 , Nghia H. Nguyen 1,2 , Changqing Zhao 1,3 , Joseph K. 

Hoang 1 , Christine Y. Chang 1 , Mingjuan Jin 1,4 , Pauline Nguyen 1 , 

Peter T. Nguyen 1,5 , Richard H. Le 1 , Michelle Q. Jin 1 , Linda 

Nguyen 6 , Lee Ann Yasukawa 7 , Jian Q. Zhang 8 , Susan C. Weber 7 , 



of Medicine, University of California San Diego, San Diego, 

CA; 3 Department of Cirrhosis, Institute of Liver Disease, Shuguang 

Hospital, Shanghai, China; 4 Department of Epidemiology and Biostatistics, 

School of Public Health, Zhejiang University, Hangzhou, 

China; 5 School of Medicine, University of Texas Medical Branch, 

Galveston, TX; 6 Stanford University, Palo Alto, CA; 7 Center for 

Clinical Informatics, Stanford University School of Medicine, Palo 

Alto, CA; 8 Chinese Hospital, San Francisco, CA 

BACKGROUND: Little is known about the natural history of 

CHC in AA .Our goal is to evaluate the rate of progression 

to cirrhosis and hepatic decompensation in a large ethnically


diverse CHC cohort. METHODS: Using ICD-9 query and manual 

chart review for all identified patients, 5,942 consecutive 

adults with CHC and known ethnicity from 3 community clinics 

and one large U.S. university center were confirmed and 

included. RESULTS: AA were older (mean age=60.3±13.4 vs. 

55±10.2, p


1850 

Prevalence of hepatitis B core antibody (anti-HBc) and 

association with hepatocellular carcinoma (HCC) in 

patients with chronic hepatitis C virus (HCV) infection in 

the United States (US) and China 

Ming Yang 1 , Elizabeth Wu 2 , Sherry Fu 2 , Huiying Rao 1 , Bo Feng 1 , 

Andy Lin 3 , Ran Fei 1 , Neehar D. Parikh 2 , Robert J. Fontana 2 , Anna 

S. Lok 2 , Lai Wei 1 ; 1 Peking University People’s Hospital, Peking 

University Hepatology Institute, Peking University Health Science 

Center, Beijing, China; 2 Division of Gastroenterology, University 

of Michigan Health System, Ann Arbor, MI; 3 The Molecular and 

Behavioral Neuroscience Institute, University of Michigan, Ann 

Arbor, MI 

Background: Evidence of previous hepatitis B virus (HBV) infection 

[i.e. detectable anti-HBc with undetectable hepatitis B 

surface antigen (HBsAg)] has been associated with increased 

risk of HCC in patients with chronic HCV infection in many 

Asian studies but this was not confirmed in some studies in the 

US. Aims: To compare the prevalence of anti-HBc between US 

and Chinese patients with chronic HCV and identify clinical 

correlates of anti-HBc positivity. Methods: Prospective study 

of 2 cohorts of chronic HCV patients at University of Michigan 

in Ann Arbor, US and Peking University People’s Hospital 

in urban Beijing and Gu’an and Kuancheng Clinics in rural 

Hebei, China. Results: A total of 1,833 patients were analyzed 

(963 US; 870 Chinese). 32.3% of US and 46.6% of 

Chinese HCV patients were anti-HBc+ and HBsAg- (p0.001). 

The prevalence of anti-HBc among patients with chronic hepatitis, 

cirrhosis, and HCC was 30.5%, 32.3% and 38.5% in 

the US (p=0.250) and 44.5%, 53.8% and 66.7% in China 

(p=0.020), respectively. Anti-HBc+ patients were older, had 

longer estimated duration of infection and were more likely to 

have a history of alcohol and tobacco use in both cohorts. US 

anti-HBc+ patients were more likely to be men and to report 

a history of injection drug use as the source of infection. In 

contrast, Chinese anti-HBc+ patients were more likely to report 

medical procedures as the source of infection. Multivariate 

analysis showed that there was no association between anti- 

HBc positivity and HCC in the US and Chinese cohorts and the 

combined cohort. Conclusions: Evidence of previous HBV infection 

among patients with chronic HCV is lower in the US cohort 

compared to the Chinese cohort. Although the prevalence of 

anti-HBc paralleled the severity of HCV-related liver disease, 

anti-HBc positivity was not an independent factor associated 

with risk of HCC in patients with chronic HCV in both US and 

Chinese cohorts. 

Characteristics of Patients with and without anti-HBc in the US 

and China 





The following authors have nothing to disclose: Ming Yang, Elizabeth Wu, Sherry 

Fu, Huiying Rao, Bo Feng, Andy Lin, Ran Fei, Neehar D. Parikh 

1851 

Wisteria floribunda agglutinin-positive Mac-2-binding 

protein predicts hepatocellular carcinoma development 

in chronic hepatitis C patients who achieved sustained 

virological response to interferon-based anti-viral therapy 

Shunsuke Sato, Hironori Tsuzura, Takuya Genda, Akihito Nagahara; 

Gastroenterology and Hepatology, Juntendo University Shizuoka 

Hospital, Izunokuni, Japan 

Objective: The achievement of a sustained virological response 

(SVR) after anti-viral therapy reduces the incidence of hepatocellular 

carcinoma (HCC) development in patients with chronic 

hepatitis C. In advances of direct acting antivirals against hepatitis 

C virus, it is expected to drastically increase the patients 

with achievement of SVR. However, HCC development in 

patients who achieved SVR is not rarely observed. Liver fibrosis 

is known as a predictor of HCC development in patients with 

achievement of SVR. Although several noninvasive methods 

evaluating liver fibrosis has been reported, wisteria floribunda 

agglutinin (WFA)-positive Mac-2-binding protein (WFA + -M2BP) 

is recently developed as a noninvasive glycobiomarker of liver 

fibrosis. The aim of this study is to evaluate WFA + -M2BP as 

a predictive marker of HCC development in chronic hepatitis 

C patients with achievement of SVR. Methods: A total 280 

patients who achieved SVR by interferon-based anti-viral therapy 

and underwent measurement of serum WFA + -M2BP level 

were enrolled. We compared its usefulness as a predictive 

marker of HCC development to other risk factors including age, 

gender, body mass index, fibrosis stage, aspartate aminotransferase, 

alanine aminotransferase, gamma-glutamyl transpeptidase, 

and alpha-fetoprotein (AFP) by using multivariate Cox 

proportional hazard analysis. Cumulative incidences of HCC 

development were evaluated using Kaplan-Meier plot analysis 

and the log-rank test. Results: Serum WFA + -M2BP level was significantly 

correlated with liver fibrosis stage (p=0.001). During 

the median follow-up time of 4.0 years, 9 of the 280 patients 

developed HCC. Multivariate cox proportional hazard analysis 

revealed that WFA + -M2BP level was an independent risk factor 

of HCC development (hazard ratio (HR): 1.167, 95% confidence 

interval (CI): 1.015-1.342, p=0.030) as well as platelet 

counts (HR: 0.682, 95%CI: 0.538-0.864, p


1852 

HCV in Semen of HIV-infected Men During Acute and 

Chronic Infection 

Samuel Turner 1,4 , Marcus Yip 1,5 , Wouter van Seggelen 1,6 , Davey 

M. Smith 2,3 , Sara Gianella 2 , Daniel S. Fierer 1 ; 1 Infectious Diseases, 

Mount Sinai School of Medicine, New York, NY; 2 Infectious 

Diseases, University of California, San Diego, San Diego, CA; 

3 Infectious Diseases, Veterans Affairs Medical Center, San Diego, 

CA; 4 James Cook University, Cairns, QLD, Australia; 5 Monash University, 

Melbourne, VIC, Australia; 6 Amsterdam Medical Center, 

Amsterdam, Netherlands 

Introduction It remains unclear how hepatitis C virus (HCV) is 

transmitted in the epidemic of sexually transmitted HCV among 

HIV-infected men who have sex with men (MSM). Our previous 

epidemiological study in New York City strongly implicated 

semen, and seminal HCV has been detected intermittently and 

at low levels during chronic HCV, but little is known about 

seminal HCV during acute HCV infection. Methods HIV-infected 

MSM with acute and chronic HCV infection were prospectively 

enrolled. Three paired semen and blood specimens 

were collected at 2-week intervals. For men with acute HCV, 

all specimens were collected within 6 months of detection of 

new HCV infection. HCV viral load (VL) was quantified using 

qRT-PCR platforms (Abbott m2000 with lower limit of quantification 

[LLOQ] 12 IU/mL for seminal plasma, and Roche 

COBAS AMPLICOR with LLOQ 43 IU/mL for blood). Results 

Paired semen and blood specimens were obtained from 33 

HIV-infected MSM (21 acute, 12 chronic HCV). Overall, seminal 

HCV was detected in 16 (27%) of 59 specimens from 

11 (33%) men (Table). Comparing men with acute or chronic 

HCV, there were no differences in either the proportion of 

semen specimens with HCV detected (21% vs 38%, respectively; 

p=0.159), or in the median seminal HCV VL (1.32 vs 

1.77 log 10 

IU/mL, respectively; p=0.163). Median blood HCV 

VL was higher among men with detectable compared to not 

detectable seminal HCV (6.36 vs 5.47 log 10 

IU/mL, respectively; 

p=0.002). Among men with low serum VL (


1854 

Modeling the probability of hepatitis C virus transmission 

in injecting drug users as a function of viral load 

Qingwen Cui 1 , Alexander Gufraind 2,3 , Basmattee Boodram 3 , 

Scott Cotler 2 , Harel Dahari 2,4 , Marian E. Major 1 ; 1 CBER/FDA, 

Alexandria, VA; 2 Department of Medicine, Loyola University, Maywood, 

IL; 3 Epidemiology & Biostatistics, University of Illinois at 

Chicago, Chicago, IL; 4 Theoretical Biology and Biophysics, Los 

Alamos National Laboratory, Los Alamos, NM 

Background: The major route of hepatitis C virus (HCV) transmission 

in the USA is through injecting drug use. It has been 

proposed that vaccines would not need to achieve sterilizing 

immunity in injecting drug user (IDU) populations in order to 

reduce disease spread, but this has not been tested empirically. 

We aimed to simulate HCV transmission through contaminated 

syringe sharing, apply this to a mathematical model to determine 

infection probabilities relative to HCV RNA and assess 

the impact of measures that reduce titers (e.g. vaccines or antivirals) 

in IDU populations. Method: HCV RNA positive (HCVpos) 

plasma was drawn into an insulin syringe and expelled. 

Negative plasma (NEG-pre) was drawn into the same syringe 

and expelled (NEG-post). Aliquots of HCV-pos, NEG-pre, and 

NEG-post were tested for RNA titer. This was repeated employing 

a water rinse of the syringe after expulsion of the HCV-pos 

sample. A binomial model was used to estimate the probability 

of infection with and without syringe rinse, assuming an RNA to 

HCV infectivity ratio of 12:1 [range=10:1-20:1]. Results: We 

found that 0.5% [range 0.05%-1.16%] of an HCV-pos sample 

could be transferred if a syringe is not rinsed or 0.07% [range 

0-0.18%] when rinsing is employed. Predicted probabilities 

of infection from syringe sharing as a function of RNA titer 

are shown in Fig.1. Risk of transmission increases 11-fold as 

titers (log10 IU/mL) rise from 2.6 to 4.0 (unrinsed) and 3.6 to 

5.0 (rinsed). Conclusions: IDUs experience a risk of transmission 

even with syringe rinsing. However, if titers are reduced 

below 2 log10 IU/mL transmission via syringe sharing could 

be prevented. Overall, a vaccine would not need to achieve 

sterilizing immunity to reduce disease spread in IDUs. Similarly, 

antiviral treatment could impact spread before sustained 

response is achieved. 

Fig1:Grey bands: Interquartile ranges based on Monte Carlo 

sensitivity analysis. 

Disclosures: 

The following authors have nothing to disclose: Qingwen Cui, Alexander 

Gufraind, Basmattee Boodram, Scott Cotler, Harel Dahari, Marian E. Major 

1855 

Heterogeneous IL28B/IFNL4 distribution and association 

of the C/TT/T haplotype with spontaneous clearance 

and less liver damage in Mexican patients with chronic 

hepatitis C virus infection 

Karina Gonzalez-Aldaco 1,2 , João Renato R. Pinho 3 , Sonia 

Roman 1,2 , Ketti G. Oliveira 3 , Nora A Fierro 1,2 , Leticia Oyakawa 3 , 

Rubia A. Santana 3 , Erika Martinez-Lopez 1,2 , Roberta Sitnik 3 , 

Arturo Panduro 1,2 ; 1 University of Guadalajara, Jalisco, Mexico; 

2 Molecular Biology in Medicine, Civil Hospital of Guadalajara, 

Guadalajara, Mexico; 3 Hospital Israelita Albert Einstein, Sao 

Paulo, Brazil 

Background & aims. Recently, genetic polymorphisms of interleukin-28B 

(IL28B) (rs12979860 C allele and rs8099917 T 

allele) and interferon-lambda 4 (IFNL4) (ss469415590 TT 

allele) have been associated with spontaneous clearance (SC) 

of hepatitis C virus (HCV) infection among distinct populations 

worldwide. However, data regarding admixed and Amerindian 

populations from Latin America are lacking. This study aimed 

to analyze the genetic admixture of the West Mexico population 

based on the distribution of the IL28B (rs12979860C>T, 

rs8099917G>T) and IFNL4 (ss469415590∆G>TT) polymorphisms, 

and the association between the IL28B/IFNL4 haplotypes 

with SC and liver damage. Methods. In a cross-sectional 

study, 711 unrelated individuals from West Mexico, including 

Amerindians (86 Nahuas/95 Huicholes) and Mestizo populations 

(32 from Villa Purificación (VP), 172 from Guadalajara, 

Jalisco and 326 from Tepic, Nayarit) were genotyped. In a 

case-control study, 234 treatment-naïve HCV-infected Mestizo 

patients (149 with chronic hepatitis C (CHC) and 85 with SC) 

were included for the association of haplotypes with SC and 

liver damage. A Real-Time PCR assay perfomed genotyping, 

and transitional elastography (FibroScan ) staged liver damage. 

Genetic relatedness was evaluated by pairwise comparisons 

(exact tests). Both, genetic distances (Fst) and haplotypes 

were inferred by using Arlequin software (version 3.1). Linkage 

disequilibrium (LD) was calculated by GDA software (version 

1.0). All subjects signed an informed consent. The study protocol 

conformed to the Declaration of Helsinki and was approved 

by the Ethical Committee. Results. Three different clusters 

(p


1856 

Pragmatic Non-invasive test thresholds for the selection 

of patients with Hepatitis C cirrhosis for treatment with 

directly acting antiviral agents. 

William M. Rosenberg 1 , Julie Parkes 1,2 ; 1 Institute for Liver and 

Digestive Health, University College London, London, United Kingdom; 

2 Public Health Sciences and Medical Statistics, University of 

Southampton, Southampton, United Kingdom 

Background: Directly acting antiviral agents can cure HCV 

infection in most patients. Their high cost has led some healthcare 

providers to restrict use to patients with established cirrhosis. 

Non-invasive tests (NIT) for liver fibrosis are being used to 

stratify patients for liver fibrosis severity. However the selection 

of appropriate thresholds of NIT for the diagnosis of cirrhosis 

in this context is contentious. Methods: We took advantage of 

a consensus meeting of UK Hepatitis C experts to determine 

thresholds for NIT to diagnose cirrhosis. An appropriate sensitivity 

for the detection of cirrhosis was agreed by consensus. 

In addition a consensus threshold for fibroscan (11.5kPa) was 

selected. Using this threshold we reviewed published literature 

to identify test thresholds of commonly used NIT yielding a 

sensitivity for detecting cirrhosis in line with the experts’ range. 

Studies identified in the recent HTA Systematic Review of NIT 

for liver fibrosis (Corssan et al.) were considered. SPSS was 

used to analyse extracted data to determine the sensitivity for 

cirrhosis associated with a Fibroscan of 11.5±0.4kPa. The 

mean sensitivity for the diagnosis of cirrhosis associated with 

this Fibroscan reading was then used to identify the corresponding 

thresholds for APRI, FIB4 and ELF using data from published 

studies of patients with HCV infection where liver histology was 

used as the reference test for cirrhosis. Results: The expert consensus 

was that no more than 10-15% of patients with cirrhosis 

should be misdiagnosed as not having cirrhosis, equivalent to 

a test sensitivity for cirrhosis of 85-90%. Thirty three studies 

reported fibroscan thresholds for cirrhosis ranging from 27kPa. Five reported a threshold of 11.5±0.4 and were 

included in the analysis, corresponding to a mean sensitivity of 

88.4%, a figure that lies within the range of sensitivities considered 

“acceptable” by the experts. The thresholds that yielded 

similar sensitivities for the diagnosis of cirrhosis were then identified 

for APRI, FIB4 and ELF and are presented in Table 1. Conclusions: 

UK CHC experts selected sensitivity for the detection 

of cirrhosis of 88% to allocate DAA treatment. Corresponding 

thresholds for widely used NIT have been identified. 

TABLE1: Thresholds for non-invasive tests for the identification of 

cirrhosis in HCV infection. 

Disclosures: 





1857 

Illegal drug use disclosure among patients with chronic 

hepatitis C infection 

Kimberly Rhodes, Sherrie M. Harrell, Victor M. Cardenas, Andres 

Duarte-Rojo; Division of Gastroenterology and Hepatology, University 

of Arkansas for Medical Sciences, Little Rock, AR 

Background: Illegal or recreational drug use (IDU) is a major 

public health problem worldwide. It impacts the prevalence 

of hepatitis C (HCV) infection, reinfection rate, adherence to 

antiviral therapy, and rate of fibrosis progression. We aimed 

to investigate disclosure of IDU among HCV patients before 

and after an intervention aiming to improve response rate. 

Methods: Consecutive new HCV patients were evaluated at 

a tertiary center. Before their visit to clinic they completed a 

questionnaire on the use of any (A) IDU, intravenous (IV) IDU, 

snorted (S) IDU, cannabis (THC) use, and other (O) IDU. During 

clinic time patients were educated about the importance of 

disclosing an accurate substance abuse history, followed by 

a verbal reinterrogation on IDU. McNemar test was used for 

statistical analysis. Results: 126 patients (50±12 years, males 

50%) were included, and 122 (97%) answered the questionnaire. 

Among the 4 that did not answer, 3 had used IDU in the 

past (2 IV-IDU, 1 only THC). Frequencies of IDU per category 

were: A-IDU 84%, IV-IDU 67%, S-IDU 55%, THC 66%, O-IDU 

29%. The rate of patients that changed their statement on IDU 

before and after education is shown in Table. Revised IDU 

reporting was statistically significant for S-IDU (p=0.01), and 

THC (p


tion of patients harboring a pre-treatment Y93H mutation. A 

total of 702 serum samples of Japanese HCV genotype 1b 

infected patients were screened in the study. The frequency of 

NS5A-Y93H was also determined by ultra-deep sequencing 

and compared by using 55 sera of patients. Possible relationships 

between clinical phenotypes and Y93H strains were 

investigated. Results: The overall success rate was 98.9% 

and the lower detection limit of HCV RNA level could be estimated 

to be almost 1 log order. The mutant strain (Y93H) 

was observed in 23.6% (164/694) patients. The lower detection 

limit of the proportion of Y93H strains could be estimated 

at 1 to 2 %. A significant positive correlation was observed 

between the proportion of Y93H strains determined by this 

assay system and the proportion determined by deep sequencing 

(r = 0.85, P 1% of 

total population). While prevalence is decreasing, the burden 

of HCV-related morbidity and mortality continues to grow. HCV 

transmission among PWIDs is the leading contributor to new 

infections with over half of infected cases reporting a history of 

injection drug use. The Serviço de Intervenção nos Comportamentos 

Aditivos e Dependências (SICAD) estimates that there 

were 14 426 (12 732 – 16 101) active PWIDs in Portugal in 

2012, and that 88% of PWIDs were HCV infected, equivalent 

to 12 695 infected PWIDs. In addition, SICAD estimates that 

16 401 individuals were enrolled in opioid substitution therapy 

(OST), and 950 652 syringes were distributed among 

PWIDs in 2013. Understanding HCV transmission dynamics 

among high-risk populations requires robust epidemiological 

data and country-specific mathematical modeling to assess 

the potential impact of improved HCV treatment strategies. 

Methods: HCV transmission was modeled using cohorts to 

track HCV incidence and prevalence among PWIDs in the 

general population, as well as active PWIDs enrolled in OST 

and/or needle exchange program (NEP). Model assumptions 

were derived from published literature and expert consensus. 

The relative impact of increasing treatment among PWID was 

considered, including the annual number needed to treat in 

order to substantially reduce the HCV-infected PWID population 

by 2030. Results: If the current transmission paradigm 

continues, HCV infected PWIDs are projected to decline 25% 

from 12 695 infected PWIDs in 2012 to 9320 HCV infected 

PWIDs in 2030. Annually treating 100 HCV-infected PWIDs 

(approximately 1% of HCV-infected PWID population in 2012) 

resulted in a 7% reduction in HCV-infected PWIDs by 2030, 

while annual treatment of 460 PWIDs (5% of 2012 population) 

resulted in a reduction of over 30% by 2030. Treating 

1400 PWIDs annually (15% of 2012 population) resulted in 

a 91% reduction in HCV-infected PWIDs by 2030; reductions 

were even greater among NEP participants (92% reduction), 

OST participants (92% reduction) and participants of both programs 

(95% reduction). Conclusions: The results show that 

treating a relatively small number of PWIDs resulted in substantial 

decreases in the HCV infected PWID population by 

2030. These data support implementation of a screening and 

treatment strategy among PWIDs; by focusing on groups with 

high transmission rates, the burden of HCV-related morbidity 

and mortality may be decreased. 

Disclosures: 


Filipe Calinas - Advisory Committees or Review Panels: Merck Sharp & Dohme, 

Roche Pharmaceuticals, Gilead sciences, AbbVie, Janssen; Consulting: Boehringer 

Ingelheim; Speaking and Teaching: Bristol Myers Squibb, Gilead Sciences, 

Janssen; Stock Shareholder: Merck Sharpe 

Chris R. Estes - Employment: Center for Disease Analysis 

Rui T. Marinho - Advisory Committees or Review Panels: Abbvie, MSD, Roche, 

BMS, Janssen, Bayer 

The following authors have nothing to disclose: Domingos Duran, Jorge Félix, 

Fernando Maltez, Luís Mendão, Graca Vilar 

1860 

Comparison of On-Treatment HCV RNA Kinetics Using 

Two Generations of the COBAS Taqman Assay 

Tania M. Welzel 1 , Stanislas Pol 2 , Patrick Marcellin 3 , Robert H. 

Hyland 4 , Deyuan Jiang 4 , Phillip S. Pang 4 , Diana M. Brainard 4 , Vincent 

Leroy 5 , Stefan Zeuzem 1 , Marc Bourlière 6 ; 1 Johann Wolfgang 

Goethe University Medical Center, Frankfurt am Main, Germany; 

2 Department of Hepatology, Université Paris-René Descartes, Paris, 

France; 3 Hôpital Beaujon, Clichy, France; 4 Gilead Sciences, Inc, 

Foster City, CA; 5 CHU de Grenoble, Grenoble, France; 6 Hôpital 

Saint Joseph, Marseilles, France 

Background: The fully automated Roche COBAS® AmpliPrep/ 

COBAS® Taqman® (version 2.0) HCV quantification test 

(CAP/CTM) with a lower limit of quantification (LLOQ) of 15


IU/mL is replacing the Roche COBAS® TaqMan® HCV test 

(version 2.0) for use with the High Pure System (CTM/HPS, 

LLOQ of 25 IU/mL) in clinical practice. Here we describe, 

through parallel testing, the HCV RNA viral decline observed 

in the SIRIUS study (NCT01965535) using the two different 

assays. Methods: Stored, frozen samples drawn during the first 

8 weeks of treatment from 154 patients in the SIRIUS trial were 

identified and analyzed in parallel using both assay platforms. 

These results were compared with each other and with the 

results initially obtained using the CTM/HPS (LLOQ=25) system 

at the time of the study conduct (between October 2013 and 

April 2014). Results: As previously reported, 97% of patients 

in this study achieved SVR12. 765 paired samples drawn 

during the first 8 weeks of treatment were analyzed. Compared 

to HCV RNA measurements obtained with the CTM/HPS 

(LLOQ=25) assay, a higher proportion of patients had quantifiable 

HCV RNA at treatment weeks 1, 2, and 4 using the new 

CAP/CTM (LLOQ=15) assay (see Table). Using the CAP/CTM 

(LLOQ=15) assay, twenty six subjects were >LLOQ at Week 4. 

Nevertheless, 25/26 (96%) of these subjects achieved SVR12 

after 12 or 24 weeks of treatment. Comparison with the data 

obtained at the time of study conduct will be presented. Conclusions: 

In this analysis, with the next generation Ampliprep sample 

preparation with the COBAS TaqMan® HCV test (version 

2.0), as many as 17% of patients had detectable HCV RNA 

at week 4; conversely, with the older High Pure System used 

to prepare samples, as few as 1% of patients had detectable 

HCV RNA at week 4. Notably, however, with either assay, the 

likelihood of a sustained viral response did not correlate with 

early virologic response. Thus, for LDV/SOF-based therapies, 

determination of on-treatment HCV RNA levels is not suitable to 

guide treatment decisions. 

Disclosures: 












Abbvie 


Sciences, Inc 

Deyuan Jiang - Employment: Gilead Sciences 



Sciences 










Squibb 

1861 

Expanded eligibility to HCV treatment with novel IFNfree 

therapies in a large real-world cohort: The German 

Hepatitis C Registry 

Dietrich Hueppe 2,6 , Stefan Mauss 2,3 , Klaus H. Boeker 2,13 , Andreas 

Schober 2,5 , Gerlinde Teuber 2,4 , Stefan Christensen 2,11 , Uwe 

Naumann 2,12 , Claus Niederau 2,10 , Stefan Zeuzem 2,8 , Michael P. 

Manns 1,2 , Thomas Berg 2,9 , Peter Buggisch 2,7 , Markus Cornberg 1,2 , 

Christoph Sarrazin 2,8 , Heiner Wedemeyer 2,1 ; 1 Gastroenterology,Hepatology 

and Endocrinology, Hannover Medical School, 

Hannover, Germany; 2 German Hepatitis C Registry, Hannover, 

Germany; 3 MVZ Düsseldorf, Düsseldorf, Germany; 4 IFS Frankfurt, 

Frankfurt, Germany; 5 Hepatologische Praxis Göttingen, Göttingen, 

Germany; 6 Hepatologische SChwerpunktpraxis Herne, Herne, 

Germany; 7 IFI Hamburg, Hamburg, Germany; 8 University of Frankfurt, 

Frankfurt, Germany; 9 University of Leipzig, Leipzig, Germany; 

10 Hospital Oberhausen, Oberhausen, Germany; 11 CIM Münster, 

Münster, Germany; 12 Praxiszentrum Kaiserdamm, Berlin, Germany; 

13 Hepatologische Praxis Hannover, Hannover, Germany 

The German Hepatitis C Registry is a non-interventional prospective 

cohort study aiming to recruit at least 10.000 HCV-infected 

patients treated with novel direct acting antivirals 

against hepatitis C. Second generation DAAs have been used 

in Germany since January 2014 and each drug was available 

immediately after EMA approval. The current registry follows 

a previous national cohort which recruited more than 37.000 

HCV patients since 2002. The aim of this analysis was to investigate 

the evolution of patient profiles considered eligible for 

antiviral therapy over a period of 13 years with different treatment 

options available. Methods: Characteristics of patients 

enrolled in the German Hepatitis C registry after approval of 

2 nd generation DAAs since February 2014 (n=2247; period 

III) were compared with patients recruited 2002-2007 (n= 

19,115; period I) and patients recruited during 2011-2012 

when 1 st generation PIs were widely prescribed in Germany 

(n=1,768; period II). Results: The mean age of patients enrolled 

increased over the last 12 years (43.0 period I vs. 47.0 years 

period II vs. 51.0 years period III) while the gender distribution 

remained largely unchanged (male gender 58.9% vs. 64.8% 

vs. 61.2%). A slight shift in HCV genotype distribution became 

evident with less genotype 3 infections in period III (20.5%) 

than in period I (24.6%) but an increase in genotype 4 (2.9% 

vs. 2.5% vs. 5.1%). The proportion of patients with liver cirrhosis 

increased from 4.5% to 24.5% in period III while less 

patients were treatment naïve in period III compared to period 

I (56.3 vs. 86.3). In 2014/15 patient characteristics differed 

largely between individuals treated with IFN-based regimens 

(n=655) vs. IFN-free DAA combination therapies (n=1,169) 

regarding age (46.3 years vs. 54.4 years; patients >70 years 

3.1% vs. 9.3%), proportion of patients with liver cirrhosis 

(15.4% vs. 37.3%), percentage of patients with platelet counts 


Gerlinde Teuber - Advisory Committees or Review Panels: MSD, Gilead Sciences, 

BMS, Abbvie; Speaking and Teaching: MSD, Gilead Sciences, Janssen-Cilag, 

BMS, Abbvie 




Uwe Naumann - Speaking and Teaching: MSD, Roche, BMS, Abbott, VIIV, Janssen, 

Boehringer Ingelheim, Gilead 

Claus Niederau - Advisory Committees or Review Panels: MSD, Gilead, BMS, 

Janssen, Abbvie; Grant/Research Support: MSD; Speaking and Teaching: MSD, 

Gilead, BMS, Janssen, Abbvie, Roche 







Novartis 























The following authors have nothing to disclose: Andreas Schober 

1862 

Evaluation of liver and spleen transient elastography in 

chronic hepatitis C patients. 

Mohamed A. Elmazaly 1 , Ayman Alsebaey 1 , Maha M. Elsabaawy 1 , 

El-Sayed S. Tharwa 1 , Hanaa M. Badran 1 , Nermine Ehsan 2 , Eman 

A. Rewisha 1 ; 1 Hepatology department, National Liver Institute, 

Shebin elkom menoufiya, Egypt; 2 Pathology, National Liver Institute, 

Shebin Elkoom, Egypt 

Background: Liver fibrosis is a common consequence of chronic 

HCV (CHC) infection. It may evolve to cirrhosis, hepatocellular 

carcinoma. Aim: is to evaluate the role of liver stiffness 

measurement (LSM), spleen stiffness measurement (SSM) and 

their combination (CLSM) using FibroScan TM in assessment of 

liver fibrosis in CHC patients. Methods: Four hundred twenty 

treatment naïve CHC patients and 40 healthy controls were 

included. Liver, renal function tests, CBC and INR were done. 

Liver biopsy was done for all of them except if contraindicated. 

Fibrosis was graded by Metavir score. Abdominal ultrasonography 

was done before the FibroScan TM and the liver 

biopsy. Transient elastography measurement was done using 

FibroScan TM in the supine position after 6-8 hours fasting. The 

patient were classified into mild fibrosis (F1-F2, n=248) and 

significant fibrosis (F3-F4, n=172) group. Results: There were 

statistically significant difference (p=0.001) between patients 

with mild fibrosis (F1-F2) versus patients with significant fibrosis 

(F3-F4) regarding; the age (35.06±8.63 vs. 43.71±7.97 

years), serum bilirubin (0.73±0.25 vs. 1.26±0.73 mg/dl), 

serum albumin (4.42±0.32 vs. 3.84±0.51 g/dl), platelets 

(206.81±50.55 vs. 140.50±53.77×10 3 /mL), platelets spleen 

ratio (1762.20±521.26 vs. 1014.64±470.27). Furthermore a 

statistical significant difference (p=0.001) was detected with 

LSM (6.57±2.62 vs. 23.04±12.15kPa), SSM (25.56±5.36 vs. 

46.19±16.29 kPa), and CLSS (32.13±7.15 vs. 69.23±25.43 

kPa) respectively. The cutoff values of significant fibrosis were 

9.15 kPa by LSM (94.8% Sensitivity, 88.3% Specificity, 84.9% 

PPV, and 96.1.8% NPV), 27.5 kPa by SSM (94.8% Sensitivity, 

70.2% specificity, 68.8% PPV, and 95.1% NPV) and 40.85 

kPa by CLSM (92.4% Sensitivity, 91.1% specificity, 87.8% 

PPV, and 94.6% NPV). Conclusion: The measurement of liver, 

spleen stiffness by FibroScan TM or their combinations are useful 

for liver fibrosis assessment. 

Disclosures: 

The following authors have nothing to disclose: Mohamed A. Elmazaly, Ayman 

Alsebaey, Maha M. Elsabaawy, El-Sayed S. Tharwa, Hanaa M. Badran, Nermine 

Ehsan, Eman A. Rewisha 

1863 

Impact of comorbidities on the benefit of DAA therapy 

in Hepatitis C 

Javier Ampuero 1 , Carlota Jimeno 1 , Angela Rojas 1 , A. Ortega 2 , 

Jose Miguel Rosales-Zabal 4 , Marta Maraver 3 , Raquel Millán 1 , 

Blanca Fombuena 1 , Ramón Morillo 5 , J. M. Navarro 4 , Raul J. 

Andrade 2 , Manuel Romero-Gomez 1 ; 1 Unit for the Clinical Management 

of Digestive Diseases, Valme University Hospital, Sevilla, 

Spain; 2 Digestive Department, Hospital Virgen de la Victoria, Malaga, 

Spain; 3 Digestive Department, Hospital Juan Ramón Jiménez, 

Huelva, Spain; 4 Digestive Department, Hospital Costa del Sol, 

Marbella, Spain; 5 Farmacology Unit, Valme University Hospital, 

Sevilla, Spain 

Background: HCV therapy achieving sustained virological 

response(SVR) decreases dramatically all-cause mortality. 

New direct acting antivirals(DAAs) are able to achieve SVR 

on more than 90% of patients. However, the benefit of the 

treatment in patients with severe comorbidities, beyond of the 

eradication of the virus, remains unknown. Aim: To explore 

the impact of comorbidities scales on decision-making according 

to HCV therapy outcomes. Methods: Prospective multicenter 

study including 104 consecutively HCV patients(≥F2 

by transient elastography) treated with DAAs from January 

2015. Basal comorbidities were evaluated by: a) Charlson 

comorbidity index(CCI) (Charlson, J Chron Dis 1987); b) 

Modified Charlson comorbidity index(mCCI) (Berkman, Ann 

Intern Med 1992); c) CirCom index (Jepsen, Gastroenterol 

2014). Adverse events were collected from the starting date 

of the treatment and were classified according to healthcare 

requirement: a) hospital admission(>24h); b) hospital observation(


including each score, to find independent variables related to 

adverse events: a) CCI [HR 2.66(95%CI 1.53-4.65);p=0.001]; 

b) mCCI [HR 2.35(95%CI 1.61-3.42);p=0.0001] and albumin 

[HR 0.99(95%CI 0.99-1.00);p=0.040]; c) CirCom [HR 

4.48(95%CI 1.98-10.14);p=0.0001] and albumin [HR 

0.997(95%CI 0.995-0.999);p=0.040]. Conclusion: HCV 

patients with higher basal comorbidities are at risk of suffering 

adverse events during first months of treatment. Charlson and 

CirCom comorbidities index appear to be important to evaluate 

the overall benefit of the HCV treatment, beyond of the 

eradication of the virus. Partients with a Charlson >3 requires 

individual analysis before starting DAA therapy. 

Disclosures: 

Jose Miguel Rosales-Zabal - Speaking and Teaching: janssen 



S.A. 

The following authors have nothing to disclose: Javier Ampuero, Carlota Jimeno, 

Angela Rojas, A. Ortega, Marta Maraver, Raquel Millán, Blanca Fombuena, 

Ramón Morillo, J. M. Navarro, Raul J. Andrade 

1864 

Perceived risk and HCV knowledge in a high risk nonbaby 

boomer population from a community based testing 

model 

Stephanie Tzarnas, Allison Brodsky, Hunana Chaudhry, Carla 

Coleman, Michelle Dougherty, Lora Magaldi, Carolyn Moy, 

Ta-Wanda Preston, Stacey B. Trooskin; Drexel University College 

of Medicine, Philadelphia, PA 

Background: Hepatitis C is the most common blood-borne 

infection in the US with a prevalence of 1.6%. CDC guidelines 

recommend high-risk individuals and individuals born between 

1945-1965 be tested for HCV. High-risk individuals may not 

be tested due to lack of access to primary care or because providers 

may not routinely assess risk factors. We aim to assess 

perceived risk for contracting HCV as well as HCV knowledge 

in a community based HCV testing program serving high risk 

individuals. Methods: Community based testing was conducted 

by two community based organizations. A brief survey was 

utilized to assess general demographic information, perceived 

risk of HCV infection, and HCV knowledge. Results: 1,587 

patients were tested from June-April 2015, 11.3% were antibody 

positive. Of the 1,587 individuals tested, 67% were born 

outside of the birth cohort. Of the 1,063 individuals tested 

outside of the birth cohort, 9.3% were HCV antibody positive. 

80% of individuals born outside of the birth cohort with reactive 

antibody test received confirmatory NAT testing. Of those 

individuals; 81% were chronically infected; only 3.2% had 

seen their PCP in the last 12 months. Notably, 97% of HCV 

antibody positive individuals outside of the birth cohort had 

histories of drug use; of this group 47% had injected drugs 

and 66% of those individuals admitted to sharing injection 

equipment. Of the HCV antibody positive individuals outside 

of the birth cohort, 75% had heard of HCV; 66% answered 

that there was a cure for HCV, and 33% believed that there 

was a vaccine for HCV. Of the non-baby boomer antibody 

positive individuals, 40% rated themselves high risk for contracting 

HCV, 15% rated themselves low risk, 11% rated themselves 

moderate risk, and 9% rated themselves not at risk for 

contracting HCV. Conclusions: Community based testing and 

linkage to care programs serving high risk individuals have 

the potential to engage individuals in subspecialty care, where 

the standard health care system may have failed to do so. 

Interestingly, many of the non-baby boomer antibody positive 

individuals rated their perceived risk for contracting HCV high, 

while few rated themselves not at risk. Community based testing 

programs are needed to ensure that high risk individuals 

outside of the 1945-1965 birth cohort receive proper HCV 

testing, treatment, and education. 

Disclosures: 

Stephanie Tzarnas - Grant/Research Support: Gilead Sciences 

Allison Brodsky - Grant/Research Support: Gilead 

Hunana Chaudhry - Grant/Research Support: Gilead Sciences 

Carla Coleman - Grant/Research Support: Gilead Sciences 

Michelle Dougherty - Employment: Opening Doors for Diverse Populations to 

Health Dispairites Research Under the NIH 1R25MD006792-01 The National 

Institute on Minority Health and Health Disparities (NIMHA) Dr. Shannon Marquez, 

PI; Grant/Research Support: Gilead Sciences 

Lora Magaldi - Grant/Research Support: Gilead Sciences 

Carolyn Moy - Grant/Research Support: Gilead 

Stacey B. Trooskin - Advisory Committees or Review Panels: Gilead Sciences; 


The following authors have nothing to disclose: Ta-Wanda Preston 

1865 

Investigation into the high prevalence of hepatitis C 

virus infection in a rural village in southwest China 

Shiqi Tao 1 , Guangyu Huang 1 , Li Wang 1,2 , Dengming He 1 , Zehui 

Yan 1 , Shitao Ding 1 , Yunjie Dan 1 , Cheng Xu 1 , Xianghua Zeng 1 , 

Xiaohong Wang 1 , Yuming Wang 1 ; 1 Department of infectious 

diseases, Southwest Hospital, Third Military Medical University, 

Chongqing, China; 2 Medical college of Guiyang, Guiyang, China 

Purpose: To clarify the changing pattern of hepatitis C virus 

(HCV) prevalence in China, with the aim of developing appropriate 

and effective strategies for the prevention and treatment 

of this significant emerging disease. Methods: The residents of 

Village M, located in southwest China, voluntarily participated 

in this study. Blood samples were obtained and anti-HCV titers 

were tested to determine the HCV status of the participants. 

For those who were anti-HCV positive, HCV RNA levels and 

genotypes were subsequently tested. HBV-related factors and 

anti-HIV titers were also tested. In addition, the methodology 

historically used to sterilize needles in the medical center used 

by the villagers was recreated to test the effectiveness of this 

procedure. Results: Anti-HCV antibody testing showed that 258 

(50.4%) of the 512 participants were anti-HCV positive. The 

majority (180/258) of anti-HCV positive participants were also 

HCV RNA positive. Genotyping based on NS5B sequences 

showed that the predominant subtype of HCV was 3b (96.0%), 

followed by 6a (2.0%) and 1b (2.0%). The recreated medical 

procedure indicated that the transmission route might have 

been inadequately sterilized needles. The HBV infection rate 

among participants was 34.4% and the HBV/HCV co-infection 

rate was 17.6%, but none of the participants were anti-HIV 

positive. Conclusions: HCV infection was found to be highly 

prevalent in a village in southwest China. A new transmission 

or prevalence pattern of HCV infection was identified that 

involved inadequately sterilized needles. Co-infection with HBV 

among the anti-HCV positive individuals was also common.


HCV molecular phylogenetic tree based on the NS5B sequences 

1866 

Hepatitis C Eradication with Sofosbuvir Leads to Significant 

Metabolic Changes 

Amilcar Morales, Manish B. Singla, Maria Sjogren, Dawn Torres; 

Gastroenterology, Walter Reed National Military Medical Center, 

Bethesda, MD 

Infection with hepatitis C can cause metabolic complications, 

including insulin resistance, hepatic steatosis and lipid abnormalities. 

The purpose of this study was to assess the effect of 

hepatitis C virus (HCV) eradication with sofosbuvir (SOF) regimens 

on glycemic and lipid tests. This is a retrospective analysis 

of HCV patients treated and cured with a SOF regimen (with 

ribavirin [RBV], interferon or simeprevir) from January 2014 to 

November 2014. Patients with HbA1c and lipid panels within 

6 months before and 6 months after therapy were identified. In 

those treated with a RBV regimen, HbA1c was obtained a minimum 

of 3 months post therapy. Medical history, demographics, 

HCV genotype, pre-therapy RNA, pre-therapy liver associated 

enzymes, pre and post-therapy hemoglobin and liver biopsies 

were included in our analysis. Patients with an increase in 

therapy for hyperlipidemia or diabetes during our study period 

were excluded. 106 patients were treated for HCV, of which 

27 patients met inclusion criteria. 66.7% were male, 37.0% 

were Caucasian, 18.5% were African American, 85.2% had 

genotype 1, and 33.3% had stage 3 or 4 fibrosis on biopsy. 

Mean age was 60.6 ± 5.2 years. Pre-treatment body-mass 

index (BMI) did not differ from post-treatment BMI (27.9 ± 4.8 

kg/m2 vs 27.4 ± 4.6 kg/m2, p = 0.662). No patients had 

an increase in their HbA1c. HbA1c significantly decreased 

with treatment of HCV (pretreatment 6.94 ± 1.05 mg/dL vs 

post-treatment 6.15 ± 0.77 mg/dL, p = 0.012). There was 

no correlation between decreases in HbA1c and changes in 

hemoglobin in patients treated with RBV. 12 patients had diabetes, 

and 2 patients had a decrease of their diabetic therapy 

after eradication. 20 patients had pre- and post-treatment lipid 

panels; there was a significant increase in LDL and total cholesterol 

during the period of observation (LDL: 92.9 ± 26.8 mg/ 

dL vs 122.8 ± 36.2 mg/dL, p = 0.005; total cholesterol (TC) 

163.2 ± 32.0 mg/dL vs 193.3 ± 45.7 mg/dL, p = 0.021). 

Our study showed a significant drop in HbA1c up to 6 months 

after eradication of HCV with a SOF regimen, which may be 

related to a decrease in inflammatory cytokines and regulation 

of insulin activation cascades. Our study is unique as most of 

our patients had clinically significant HgA1C prior to HCV 

therapy. Increased LDL and total cholesterol are likely due to 

HCV infectious mechanism; lipid alterations are frequently 

seen post therapy. This study suggests that physicians treating 

HCV patients should reassess preventive medicine measures 

after therapy, and the benefits of eradicating HCV may extend 

beyond eliminating the effects of chronic liver inflammation. 

Disclosures: 

Maria Sjogren - Speaking and Teaching: Gilead, Bristol Myers 

The following authors have nothing to disclose: Amilcar Morales, Manish B. 

Singla, Dawn Torres 

Disclosures: 

The following authors have nothing to disclose: Shiqi Tao, Guangyu Huang, Li 

Wang, Dengming He, Zehui Yan, Shitao Ding, Yunjie Dan, Cheng Xu, Xianghua 

Zeng, Xiaohong Wang, Yuming Wang 

1867 

Hepatitis C and Renal Disease: Differences in Patient 

Characteristics and Clinical Outcomes in the United 

States 

Senaka Peter 1 , Craig Solid 2 , Tanya Bovitz 2 , Haifeng Guo 2 , Allan 

J. Collins 2 , Jean Marie Arduino 1 ; 1 Merck & Co., Inc, Kenilworth, 

NJ; 2 Chronic Disease Research Group, Minneapolis, MN 

Purpose: To describe characteristics and outcomes of US commercially-insured 

patients with hepatitis C virus (HCV) infection 

with and without renal impairment. Methods: In a cross-sectional 

analysis, we identified an HCV cohort of patients aged 

20 to 64 using Truven MarketScan claims database based 

on ICD-9 diagnosis codes in 2011. Among this cohort, we 

identified those with evidence of chronic kidney disease (CKD) 

and end-stage renal disease (ESRD) by ICD-9 and CPT codes. 

Adjusted logistic regression was used to estimate the odds of 

clinical outcomes during a 1-year follow-up period. Results: 

Of the 35,965 HCV patients identified, when compared with 

HCV patients without evidence of renal impairment, those with 

renal impairment were older, more often male, and had a 

higher prevalence of most comorbidities, including hepatitis 

B virus, cirrhosis, decompensated cirrhosis, and hepatocellular 

carcinoma. Renal disease (either non-ESRD CKD or ESRD) 

was associated with a significant increase in the rates of most 

clinical outcomes studied, including heart failure (HF), sepsis, 

pneumonia, stroke, AMI, and severe anemia with the need for 

blood transfusion (Figure). When repeated using Medicare 

data and HCV patients aged 65 years and older, similar results 

were obtained (data not shown). Conclusions: Our findings 

suggest that HCV patients with concurrent renal disease have a 

significantly higher disease burden and likelihood of negative 

health outcomes. These findings are of particular interest given 

the efficacy of new HCV treatment options. Future studies can 

assess how these new treatments may influence the likelihood 

of adverse outcomes among HCV patients with renal impairment.


majority were male (HBV DC 82%, HBV HCC 86%, HCV DC 

76%, HCV HCC 81%). The number of incident hospitalizations 

for HCV DC and HCV HCC has increased over time (Figure). 

In contrast, incident hospitalizations for HBV DC and HBV HCC 

have remained stable (Figure). Over the study period, estimated 

uptake of antiviral therapy has increased for HBV (2% to 

5%), but remained low for HCV (1-2% per annum). Conclusion 

The burden of HCV-related hospitalizations due to ESLD has 

increased markedly. Planned linkage to individual-level data 

on HBV and HCV antiviral therapy prescriptions should provide 

further insights into the contrasting pattern in ESLD burden. 

Population level monitoring of HCV ESLD burden will be particularly 

crucial to evaluate the impact of interferon-free regimens. 

Disclosures: 

Senaka Peter - Employment: Merck 

Allan J. Collins - Advisory Committees or Review Panels: KDIGO, Amgen, IKEA-J, 

NKF KDPN, Hospira; Board Membership: Kidney Health Australia; Consulting: 

Amgen, NxStage, Keryx, Hospira, Relypsa, Bayer, ZS Pharma; Employment: 

Hennepin Healthcare System; Grant/Research Support: Amgen, Merck, 

NxStage, ZS Pharma, Keryx, Peer Kidney Care Initiative; Speaking and Teaching: 

Hospira, Amgen 


The following authors have nothing to disclose: Craig Solid, Tanya Bovitz, Haifeng 

Guo 

1868 

Trends in end-stage liver disease among people notified 

with HBV or HCV in New South Wales, Australia: 2000- 

2014 

Reem K. Waziry 1 , Jason Grebely 1 , Janaki Amin 1 , Maryam Alavi 1 , 

Behzad Hajarizadeh 1 , Jacob George 2 , Gail Matthews 1 , Matthew 

Law 1 , Gregory Dore 1 ; 1 The Kirby institute, University of New South 

Wales, Sydney, NSW, Australia; 2 Storr Liver Unit, Westmead Millennium 

Institute for Medical Research and Westmead Hospital, 

University of Sydney, Westmead, NSW, Australia, Sydney, NSW, 

Australia 

Study aims To assess trends in hospitalizations for end-stage 

liver disease (ESLD); (decompensated cirrhosis (DC) and hepatocellular 

carcinoma (HCC)) among people notified with HBV 

or HCV in New South Wales (NSW), Australia. Methods 

HBV and HCV cases notified to the NSW Health Department 

between 1993 and 2012 were linked to data on hospitalizations 

(2000-2014). Time trends in hospitalizations (incident 

and total) due to DC (including ascites, esophageal varices 

with bleeding, hepatic failure, alcoholic hepatic failure, and 

alcoholic liver cirrhosis) and HCC were evaluated [International 

Classification of Diseases (ICD 10) coding]. Results 

Between 1993 and 2012, a total of 151,307 individuals were 

notified with HBV (n=54,399), HCV (n=93,099), or HBV/ 

HCV (n=3,809). From 2000 to 2014, there were 24,130 

(HBV=2,346; HCV=20,713, HBV/HCV=1,071) and 7,044 

(HBV=2,583; HCV=4,180, HBV/HCV =281) hospitalizations 

for DC and HCC, respectively. Among all hospitalizations 

for ESLD, the number of incident hospitalizations was 

6,031 (HBV=907, HCV=4,848, HBV/HCV=276) for DC and 

2,055 (HBV=732, HCV=1,244, HBV/HCV=79) for HCC. 

Median age at admission with ESLD was 57 (IQR=12) and 

58 (IQR=20) for HBV DC, 61 (IQR=16) for HBV HCC, and 51 

(IQR=11) for HCV DC and 58 (IQR=13) for HCV HCC .The 

Disclosures: 



Jacob George - Advisory Committees or Review Panels: Roche, BMS, MSD, 

Gilead, Janssen, Abbvie; Grant/Research Support: MSD 

Gail Matthews - Advisory Committees or Review Panels: gilead; Consulting: Viiv; 

Grant/Research Support: Gilead Sciences, janssen; Speaking and Teaching: 

BMS, MSD 

Matthew Law - Grant/Research Support: Boehringer Ingelhiem, Gilead Sciences, 

Merck Sharp & Dohme, Bristol-Myers Squibb, Janssen-Cilag, ViiV HealthCare 




The following authors have nothing to disclose: Reem K. Waziry, Janaki Amin, 

Maryam Alavi, Behzad Hajarizadeh 

1869 

The influence of Hepatitis C virus infection and its clearance 

on the serum lipid profile and glucose level 

Yinping Li, Xiaomei Wang, Hongqin Xu, Ruihong Wu, Xiumei Chi, 

Xiuzhu Gao, Yu Pan, Junqi Niu; Hepatology, The First Hospital of 

Jilin University, Changchun, China 

Chronic hepatitis C (CHC) is associated with a disturbance of 

lipid metabolism and glucose intolerance, and the clearance 

of HCV may be followed by a decrease in serum total cholesterol 

(TC), triglyceride (TG) and insulin resistance to adverse 

levels. The present study was designed to determine the impact 

of CHC and its treatment on circulating lipids and glucose 

levels. A total of 748 patients with HCV infection were retrospectively 

enrolled, Fasting serum total cholesterol (TC), triglyceride 

(TG) and blood glucose (FBG) levels in these patients 

were compared with 664 healthy individuals. Serum TC, TG 

and systematic glucose metabolism were measured in 183 

patients with chronic hepatitis C receiving interferon α-2b plus 

ribavirin at baseline, at the end of treatment, and at week 24 

post-treatment. We analysed systemic glucose metabolism in 

terms of the following indices: fasting insulin (FINS), C-peptide 

(FCP), homeostasis model assessment for insulin resistance 

(HOMA-IR) and beta-cell function (HOMA-β) and the insulin


sensitivity index (ISI).Patients with HCV infection had significantly 

lower TC and TG than healthy controls. This difference 

remained significant when adjusted for sex, age and BMI. 

During the antiviral treatment, we found a significant reduction 

of TC levels independently from the outcome of treatment, 

but this rebounded to baseline in patients achieving a SVR 

(4.51-4.45mmol/l, P=0.509) and stayed below baseline in 

patients without SVR (4.39-4.72mmol/l, P=0.004). Serum TG 

levels were also significantly elevated in both groups at the end 

of treatment, and persisted until week 24 post-treatment only 

in the patients with SVR. In sustained responders, fasting insulin 

(P=0.007), C-peptide (P


guidelines during the time of this study (including prioritizing 

RX among those with advanced liver disease and non-genotype 

1). Anticipation of more efficacious therapy (1 st generation 

DAAs in development > 2007; and release of 2 nd generation 

DAAs >2013) also influenced RX trends. 

Disclosures: 

The following authors have nothing to disclose: Carla V. Rodriguez, Kevin Rubenstein, 

Haihong Hu, Michael A. Horberg 

1872 

Can Hepatitis C be eliminated from a high disease burden 

country like Pakistan? 

Saeed Hamid 1 , Huma Qureshi 2 , Asad A. Chaudhry 3 , Zaigham 

Abbas 4 , Altaf Alam 5 , Altaf Baqir 6 , Javed I. Farooqi 7 , Muhammad 

S. Memon 8 , Arif A. Nawaz 9 , Homie Razavi 10 , Devin M. Razavi-Shearer 

11 , Amjad Salamat 12 , Masood Siddiq 13 , Arif Siddiqi 14 , 

Ghias Un Nabi Tayyab 15 , Muazzam Uddin 16 , Aasim Yusuf 17 , 

Bader F. Zuberi 18 , Aamir G. Khan 19 , Syed M. Jafri 1 ; 1 Medicine, 

Aga Khan University, Karachi, Pakistan; 2 Pakistan Medical 

Research Council, Karachi, Pakistan; 3 Liver Foundation, Gujranwala, 

Pakistan; 4 Sindh Institute of Urology and Transplantation, 

Karachi, Pakistan; 5 Skeikh Zayed Medical Institute, Lahore, Pakistan; 

6 Nishtar Hospital, Multan, Pakistan; 7 Post Graduate Medical 

Institute, Peshawer, Pakistan; 8 AIMS, Hyderabad, Pakistan; 

9 Fatima Memorial Hospital, Lahore, Pakistan; 10 Center for Disease 

Analysis, Denver, CO; 11 Center for Disease Analysis, Denver, Pakistan; 

12 CMH, Rawalpindi, Pakistan; 13 Fauji Foundation Hospital, 

Rawalpindi, Pakistan; 14 Allama Iqbal Medcial College, Lahore, 

Pakistan; 15 PGMI, Lahore, Pakistan; 16 Bolan Medcial College, 

Quetta, Pakistan; 17 Shaukat Khanum Hospital, Lahore, Pakistan; 

18 Dow Medical College, Karachi, Pakistan; 19 Khyber Teaching 

Hospital, Peshawer, Pakistan 

Background: Pakistan has the highest number of individuals 

with active hepatitis C virus (HCV) infection of all countries 

except China, but unlike China, the total number of infections 

in Pakistan is not declining. An estimated 4.2% of the general 

population or 7,192, 900 persons were infected with HCV in 

2014. Therefore, it is critical to assess the impact of strategies 

that can tackle the ongoing transmission and manage the HCV 

disease burden. Methods: Using a Markov model, the progression 

of the HCV infected population was quantified and 

the impact of different intervention strategies (prevention, treatment 

and screening) was measured. The inputs were gathered 

through a review of local and international publications, analysis 

of local data, and discussions with an expert panel. Findings: 

Elimination of HCV in 15 years is only feasible through 

a combination of aggressive prevention, treatment and screening. 

An estimated 230,000 new infections occur annually. 

While 85,000 individuals are treated each year, the total number 

of HCV infections is projected to increase to 7,529,200 

by 2030. Contaminated medical equipment remains one of 

the main risk factors, and education campaigns combined with 

wide-scale availability of sterile equipment could reduce the 

new infections by 50%. In addition, increasing treatment to 

510,000 patients each year with high sustained viral response 

therapies would reduce the total number of HCV infections to 

less than 700,000 by 2030. Lower number of infections in 15 

years would require the treatment of elderly and youth as well. 

Implementation of this strategy would require an increase in 

screening from 100,000 newly diagnosed cases to 600,000 

annually. Conclusions: Elimination of HCV in Pakistan in 15 

years is feasible but requires wide scale education, availability 

of sterile equipment, screening and treatment. This strategy 

would avert 220,000 liverrelated deaths and 116,000 new 

hepatocellular carcinoma cases within 15 years. 

Disclosures: 


The following authors have nothing to disclose: Saeed Hamid, Huma Qureshi, 

Asad A. Chaudhry, Zaigham Abbas, Altaf Alam, Altaf Baqir, Javed I. Farooqi, 

Muhammad S. Memon, Arif A. Nawaz, Devin M. Razavi-Shearer, Amjad 

Salamat, Masood Siddiq, Arif Siddiqi, Ghias Un Nabi Tayyab, Muazzam 

Uddin, Aasim Yusuf, Bader F. Zuberi, Aamir G. Khan, Syed M. Jafri 

1873 

Dynamic change of α-fetoprotein, platelet counts and 

aminotransferase-to-platelet ratio index (APRI) Predict 

Hepatocellular Carcinoma in chronic hepatitis C patients 

after antiviral therapy 

Cheng-Kun Wu, Kuo-Chin Chang, Po-Lin Tseng, Sheng-Nan Lu, 

Chien-Hung Chen, Jing-Houng Wang, Chuan-Mo Lee, Ming-Tsung 

Lin, Chao-Hung Hung, Yi-Hao Yen, Tsung-Hui Hu; Division of Hepato-Gastroenterology, 

Department of Internal Medicine, Kaohsiung 

Chang Gung Memorial Hospital, Taiwan, Kaohsiung City, Taiwan 

Background : Some patients who achieved viral eradication 

still developed HCC in the future. It is very important to 

accurately stratify the risks of HCC development in both SVR 

and non-SVR groups. Less is known about the role of dynamic 

change of serum markers on influence of HCC development. 

Aims: To clarify the relationship between the dynamic change 

of serum biomarkers and HCC risks Methods: We conducted 

a large-scale, long-term cohort study with 2405 CHC patients 

who have been treated with interferon (IFN) or pegylated interferon 

(peg-IFN) plus ribavirin therapies enrolled. After exclusion 

of patient with HCC development before treatment or 

within 6 months after the end of therapy, loss to follow-up and 

who didn’t have complete laboratory data, 1903 patients were 

finally assessed for analysis. The collection of pre-treatment 

hematological and biochemical data was completed before 

IFN-based therapy. Post-treatment hematological and biochemical 

data were collected at least 1 year after IFN-based therapy 

and to the date of latest follow-up if possible. AFP data were 

obtained more than 6 months prior to HCC development to 

exclude HCC-related AFP elevation. Results: HCC developed 

in 142 patients during follow-up. The entire cohort patients 

had decreased AFP after IFN-based therapy irrespective of 

viral eradication. Significantly decreased APRI was observed 

in patients with SVR and non-SVR groups. The SVR patients 

had increased platelet counts after antiviral therapy, whereas 

non-SVR patients continued to have decreased platelet counts 

during follow-up. On multivariate analysis, older age, liver cirrhosis, 

lower hemoglobin level, non-SVR status, higher pre- and 

post-treatment AFP (^15 ng/ml) and higher post-treatment APRI 

(≥0.7) were significant risk factors for HCC. Risk factors for 

HCC among patients with SVR were older age, liver cirrhosis 

and higher pre- and post-treatment AFP; among those without 

SVR, the risk factors were liver cirrhosis, lower hemoglobin 

level, lower pre-treatment platelet


1874 

Linkage to Care and Treatment of Hepatitis C in a Federally 

Qualified Health Center in New Haven, Connecticut 

Izona Bock 1 , Mina A. Garcia 1 , Robert Bruce 1,2 ; 1 Infectious Diseases, 

Cornell Scott Hill Health Center, New Haven, CT; 2 Yale 

Center for Clinical Investigation Scholar, Yale University, New 

Haven, CT 

Introduction: Approximately 2.7-3.9 million persons are 

infected with hepatitis C in the United States. The current standard 

therapy of hepatitis C is revolutionary given the high 

cure rates, short treatment duration, and relatively mild side 

effects. Our Federally Qualified Health Center (FQHC) serves 

approximately 39,000 underserved patients in New Haven 

County and provides treatment of substance abuse and chronic 

hepatitis C. The goal of this study was to develop a hepatitis 

C treatment cascade chart by determining the rates of linkage 

to care and treatment of patients with chronic hepatitis 

C in our center. Methods: With use of the electronic health 

record, we retrieved a list of patients coded with hepatitis C 

between 2013 and 2015. By performing a chart review, we 

determined the number evaluated specifically for hepatitis C 

treatment, those who were started on therapy, and the number 

who achieved virologic suppression. Results: Of the 7716 

patients screened for hepatitis C between 2013 and 2015, 

786 patients (10.2%) were identified as having hepatitis C. 

32% (250) of these patients were evaluated by a provider who 

treats hepatitis C. Ultimately, 10.4% (82) of patients started 

therapy and 6.8% (54) of those were found to have a hepatitis 

C viral load


between first and last LSM was 51 months (4-106). Median 

basal LSM was similar in treated and nontreated or between 

SVR and N-SVR. Fibrosis regression was observed in treated - 

49,2% vs NT-36,8% (p=0,30), SVR – 71% vs N-SVR- 26,7% 

(p=0.002). Analyzing the final LSM, the SVR group had a 

significant increase of the number of patients with stage < F2 

(plus 35,6%, p =0.001), in comparison to N-SVR (minus 6.4%, 

p ns). During follow-up, a significant decrease of median LSM 

was observed in SVR (from 6.55 to 5.55 kPa, p


nificantly affects the long-term natural history of this process 

even in low fibrosis groups. Median monitoring intervals of 3 

years is suggested to allocate fibrosis and is crucial in non-SVR 

& non-Tx patients with F0-F2. 

Disclosures: 

The following authors have nothing to disclose: Shaul Yaari, Orit Pappo, Saleh 

N. Daher, Ariel Benson, Nadav Ilani, Tova Goldberg-Clein, Muhammad Massarwa, 

Johnny Amer, Rifaat Safadi 

1879 

Evaluation of liver and spleen transient elastography in 

the diagnosis of esophageal varices 

Ayman Alsebaey 1 , Mohamed A. Elmazaly 1 , Maha M. Elsabaawy 1 , 

El-Sayed S. Tharwa 1 , Hanaa M. Badran 1 , Nermine Ehsan 2 , Eman 

A. Rewisha 1 ; 1 Hepatology department, National Liver Institute, 

Shebin elkom menoufiya, Egypt; 2 Pathology, National Liver Institute, 

Shebin Elkoom, Egypt 

Background: Portal hypertension with subsequent esophageal 

varices (EVs) development is a common complication of HCV 

related cirrhosis. Aim: is to evaluate the role of liver stiffness 

measurement (LSM), spleen stiffness measurement (SSM) and 

their combination (CLSM) using FibroScan TM in diagnosis of 

EVs. Methods: One hundred sixty five HCV related F3-4 Metavir 

score fibrosis were included. Liver, renal function tests, CBC, 

INR and abdominal ultrasonography were done before the 

FibroScan TM . Transient elastography measurement was done 

using FibroScan TM in the supine position after 6-8 hours fasting 

followed by diagnostic esophageogastroscopy. Varices 

were classified into none (n= 110), small (n =30) and large (n 

=25). Results: Patients with varices had higher serum bilirubin 

(1.68±0.82 vs. 1.00±0.55 mg/dl) and lower platelet count 

(105.09±31.34vs. 161.21±52.97 ×10 3 /mL) that patients 

without varices (p=0.001). The patients with varices had statistically 

significant (p=0.001) higher platelets spleen ratio 

(671.14±258.89 vs 1215.41±445.58), LSM (31.93±13.29 

vs. 17.55±6.53 kPa), SSM (62.85±12.71 vs. 36.94±8.83 

kPa) and CLSS (94.78±20.98 vs. 54.49±12.84 kPa) than 

patients without varices. In patients with small and large varices 

LSM was comparable (30.84±12.69 vs 33.64±13.97 kPa; 

p=0.391) but a statistically significant difference was detected 

with SSM (59.92±13.47 vs. 66.98±8.67 kPa; p=0.031) and 

CLSS (90.76±21.76 vs 100.62±19.00 kPa; p=0.043). With 

a cutoff of 20.4 kPa LSM (81.0% sensitivity, 71.8% specificity, 

52.3% PPV, 90.8% NPV, 74.3% accuracy), 43.2 kPa SSM 

(92.9% sensitivity, 84% specificity, 69.6% PPV, 96.9% NPV, 

86.8% accuracy) and 59.3 kPa CLSS (95.2% sensitivity, 70% 

specificity, 54.8% PPV, 97.5% NPV, 76.9% accuracy) esophageal 

varices can be detected. Conclusion: The measurement of 

liver, spleen stiffness by FibroScan TM or their combinations are 

useful for esophageal varices diagnosis. 

Disclosures: 

The following authors have nothing to disclose: Ayman Alsebaey, Mohamed A. 

Elmazaly, Maha M. Elsabaawy, El-Sayed S. Tharwa, Hanaa M. Badran, Nermine 

Ehsan, Eman A. Rewisha 

1880 

Self-reported Alcohol Use at Time of HCV Antibody 

Screening for Individuals Tested in Community Settings 

Michelle Dougherty, Allison Brodsky, Hunana Chaudhry, Carla 

Coleman, Lora Magaldi, Carolyn Moy, Ta-Wanda Preston, Stephanie 

Tzarnas, Stacey B. Trooskin; Drexel University College of 

Medicine, Philadelphia, PA 

Background: According to AASLD/IDSA, individuals with HCV 

should avoid alcohol. Although there is no safe level of alcohol 

consumption, those who consume three or more drinks daily 

are at a higher risk for accelerated liver damage. We aim 

to assess alcohol use at time of HCV antibody screening for 

those tested in community based settings. Methods: A survey 

was administered to individuals receiving point of care HCV 

testing in community based settings. Data was collected from 

June, 2014 to April, 2015. Results: Information about alcohol 

consumption was collected from 1,254 individuals who were 

tested in community based settings. Of these 1,254 individuals, 

11.80% (n=148) tested positive for HCV antibody. 29% 

of those who tested positive for HCV antibody reported current 

alcohol consumption, compared to 35.7% of individuals who 

had no antibody detected. Of the anti-HCV positive individuals 

who answered follow up questions regarding alcohol consumption 

(n=41), 36.6% reported drinking four or more times per 

week, compared to 14.6% of anti-HCV negative individuals 

who answered the same follow up questions regarding alcohol 

consumption (n=417) [p=0.0003]. Additionally, of the anti- 

HCV positive individuals who answered the follow up questions, 

17.1% reported consuming seven or more drinks when 

they drink, compared to 5.3% of anti-HCV negative individuals 

[p=0.0031]. Conclusion: Although individuals who tested 

negative for HCV in the community were more likely to report 

that they drank alcohol than those who tested positive for antibody 

to HCV, the quantity and frequency of alcohol use was 

higher among anti-HCV positive individuals. Because alcohol 

use is associated with accelerated liver damage, these findings 

underscore the need for community outreach, education and 

HCV testing programs. 

Disclosures: 

Michelle Dougherty - Employment: Opening Doors for Diverse Populations to 

Health Dispairites Research Under the NIH 1R25MD006792-01 The National 

Institute on Minority Health and Health Disparities (NIMHA) Dr. Shannon Marquez, 

PI; Grant/Research Support: Gilead Sciences 

Allison Brodsky - Grant/Research Support: Gilead 

Hunana Chaudhry - Grant/Research Support: Gilead Sciences 

Carla Coleman - Grant/Research Support: Gilead Sciences 

Lora Magaldi - Grant/Research Support: Gilead Sciences 

Carolyn Moy - Grant/Research Support: Gilead 

Ta-Wanda Preston - Grant/Research Support: Gilead Sciences 

Stephanie Tzarnas - Grant/Research Support: Gilead Sciences 

Stacey B. Trooskin - Advisory Committees or Review Panels: Gilead Sciences; 


1881 

Rate and Predictors of Serum HCV-RNA >6 Million IU/ 

mL in Chronic Hepatitis C Patients 

Pablo Barreiro 2 , Pablo Labarga 1 , Jose V. Fernandez-Montero 4 , 

Carmen de Mendoza 3 , Laura Benitez 3 , Jose M Peña 2 , Vincent 

Soriano 2 ; 1 La Luz Clinic, Madrid, Spain; 2 La Paz University Hospital 

& IdiPAZ, Madrid, Spain; 3 Internal Medicine, Puerta de Hierro 

Research Institute, Majadahonda, Spain; 4 University Hospital 

Crosshouse, Kilmarnock, United Kingdom 

Background: Baseline serum HCV-RNA predicts sustained 

virological response in chronic hepatitis C patients treated 

with both peginterferon-ribavirin and more recently with some 

direct-acting antiviral regimens. Whereas thresholds around 

800,000 IU/mL split out good and poor responders to interferon-based 

regimens, a greater threshold at 6,000,000 IU/ 

mL has been propose to discriminate treatment outcomes on 

sofosbuvir-ledipasvir. In comparison with the general population, 

immunosuppressed individuals exhibit greater viral load 

values. Methods: Serum HCV-RNA values recorded from all 

chronic hepatitis C patients consecutively attended at our clinic 

during the last decade were analyzed. The COBAS TaqMan 

HCV test v2.0 with the high pure system was used, with a


lower limit of quantification of 25 IU/mL. Results: A total of 

816 individuals with detectable serum HCV-RNA were identified. 

The main characteristics of this population were as follows: 

mean age: 48.6±6.9 years-old; male gender 73.4%; 

mean ALT: 82.6±70.9 IU/L; mean HCV-RNA: 6.02±0.75 log 

IU/mL; HCV genotypes 1 or 4: 80.6%; mean liver stiffness: 

11.3±10.4 kPa; advanced liver fibrosis (>9.5 kPa): 34.9%; 

IL28B-CC: 35.4%. HIV coinfection was found in 78.7% (mean 

CD4 count: 496±295 cells/uL; HAART: 91%). Overall, 127 

(15.6%) had serum HCV-RNA values >6 million IU/mL. This 

high viremia was found in 18.2% of HIV-positive versus 5.7% 

of HIV-negative (p6 million IU/mL were as follows: mean age 

48.2±5.2 years-old; male gender 78.7%, HCV genotypes 1-4: 

87.6%; advanced liver fibrosis 33.6%; IL28B-CC 41.1%; and 

HIV pos 92.1%. In multivariate analysis, serum HCV-RNA >6 

million IU/mL was only significantly associated with HIV coinfection 

(OR: 4.03; 95% CI: 1.98-8.19, p6 million IU/mL is overall seen 

in 6% of chronic hepatitis C patients, being significantly more 

frequent in patients with HIV coinfection (18%) and/or with 

HCV genotypes 1/4 (10%). 

Disclosures: 

The following authors have nothing to disclose: Pablo Barreiro, Pablo Labarga, 

Jose V. Fernandez-Montero, Carmen de Mendoza, Laura Benitez, Jose M Peña, 

Vincent Soriano 

1882 

Acute hepatitis and reactivation of hepatitis C virus in 

cancer patients receiving systemic chemotherapy 

Hae Lim Lee, Si Hyun Bae, Hyun Yang, Jeong Won Jang, Jong 

Young Choi, Seung Kew Yoon; the catholic university of Korea, 


Introduction : Reactivation of hepatitis C virus (HCV) is less 

common and only a few cases with severe consequences have 

been reported compared to HBV infection when it comes to 

receiving systemic chemotherapy. Many reported studies suggested 

that HCV reactivation was associated with chemotherapy 

containing rituximab and/or steroids. This retrospective 

study was aim to characterize acute exacerbation of HCV 

infection in cancer patients receiving systemic chemotherapy. 

Patients and methods : A total of 102 patients reviewed from 

January 1, 2008 to March 1, 2015 at Seoul St. Mary’s hospital, 

positive for anti-HCV antibody and receiving systemic chemotherapy 

for cancer except hepatocellular carcinoma, were 

included. 7 patients who lack HCV RNA titers were excluded. 

Hepatitis was defined as over three fold increase in ALT level. 

HCV reactivation was defined as reappearance of RNA which 

was previously undetected or resolved or RNA increasd over 

10 fold compared with the baseline level. Results : Among the 

95 patients with HCV infection and cancer, 53 patients (56%) 

showed hepatitis of all cause. 23(43%) of the 53 patients had 

hematological tumors and these were DLBCL(6, 11%), acute 

leukemia(n=11, 20%) and others(n=6, 11%). 14(33%) of the 

42 patients without hepatitis had hematological tumor. Neither 

severe hepatitis nor hepatic decompensation was noted. 32 

patients had RNA level determination before and after chemotherapy. 

HCV reactivation was noted in 10 patients (31%). 

Among them, no patient (0/10) with acute leukemia regardless 

of stem cell transplantation was identified to have HCV 

reactivation. In univariate analysis for HCV reactivation, no 

statistically significant factor was found. In 4 of 10 patients, 

hepatitis was documented in the period of HCV reactivation. 2 

patients were diagnosed as DLBCL and had received R-CHOP 

for chemotherapy regimen. Others were breast cancer and 

malignant melanoma. Conclusion : Cancer patients with HCV 

infection can be generally treated with systemic chemotherapy. 

Although systemic chemotherapy can induce hepatitis and HCV 

reactivation, no severe hepatitis or hepatic decompensation 

was documented, thus it might have less clinical significance 

compared to HBV infection. 

Disclosures: 

The following authors have nothing to disclose: Hae Lim Lee, Si Hyun Bae, Hyun 

Yang, Jeong Won Jang, Jong Young Choi, Seung Kew Yoon 

1883 

Healthcare Utilization and Costs among Hepatitis C 

Patients with and without Impaired Renal Function in 

the United States 

Senaka Peter 1 , Craig Solid 2 , Tanya Bovitz 2 , Haifeng Guo 2 , Allan 

J. Collins 2 , Jean Marie Arduino 1 ; 1 Merck & Co., Inc., Kenilworth, 

NJ; 2 Chronic Disease Research Group, Minneapolis, MN 

Purpose: To compare rates of healthcare utilization and costs 

among US commercially-insured patients with hepatitis C virus 

(HCV) infection with and without renal impairment. Methods: 

This cross-sectional analysis used Truven MarketScan claims 

database to identify an HCV cohort of patients aged 20 to 

64 based on ICD-9 diagnosis codes in 2011. Within the 

cohort, we identified those with evidence of chronic kidney 

disease (CKD) or end-stage renal disease (ESRD) by ICD-9 

codes. All-cause healthcare utilization (HCU) and medical 

costs for all health encounters excluding pharmacy during 

a 1-year follow-up period were evaluated. The subset of allcause 

HCU and costs related to HCV were identified as those 

claims with a principal diagnosis code for HCV or HCV-related 

liver disease. Generalized linear models, adjusted for 

patient characteristics and baseline comorbidities, were used 

to test for differences in costs. Results: A total of 35,965 HCV 

patients (mean age= 53.2 years; 63% male) were identified 

in 2011. All-cause and HCV-related HCU in the inpatient and 

outpatient settings were substantially higher for HCV patients 

with non-ESRD CKD or ESRD compared to HCV patients with 

no evidence of renal impairment during the 2012 follow-up 

year (Table). Compared to patients without evidence of renal 

impairment, all-cause and HCV-related medical costs (Table) 

were also significantly higher for those with renal disease, as 

demonstrated by adjusted rate ratios of 1.74 and 2.53 (both 

p


Disclosures: 

Senaka Peter - Employment: Merck 

Allan J. Collins - Advisory Committees or Review Panels: KDIGO, Amgen, IKEA-J, 

NKF KDPN, Hospira; Board Membership: Kidney Health Australia; Consulting: 

Amgen, NxStage, Keryx, Hospira, Relypsa, Bayer, ZS Pharma; Employment: 

Hennepin Healthcare System; Grant/Research Support: Amgen, Merck, 

NxStage, ZS Pharma, Keryx, Peer Kidney Care Initiative; Speaking and Teaching: 

Hospira, Amgen 


The following authors have nothing to disclose: Craig Solid, Tanya Bovitz, Haifeng 

Guo 

1884 

Does pregnancy protect against fibrosis or promote SVR 

in HCV infected women? 

Christine Pocha 1 , Pascal Benkert 2 , Salome A. Keller 1 , Beat Müllhaupt 

3 , David Semela 4 , Markus H. Heim 5 , Darius Moradpour 6 , 

Andreas Cerny 7 , Raffaele Malinverni 8 , Francesco Negro 9 , 

Jean-Francois Dufour 1 ; 1 Hepatology, University Clinic for visceral 

Surgery and Medicine,, University Hospital Bern, Bern, Switzerland; 

2 Clinical Trial Unit, University of Basel, Basel, Switzerland; 

3 Gastroenterology and Hepatology, University of Zürich, Zürich, 

Switzerland; 4 Gastroenterology and Hepatology, Kantonsspital 

St. Gallen, St. Gallen, Switzerland; 5 Gastroenterology and Hepatology, 

University Hospital Basel, Basel, Switzerland; 6 Gastroenterology 

and Hepatology, University of Lausanne, Lausanne, 

Switzerland; 7 Internal Medicine, Clinica Luganese Moncucco, 

Lugano, Switzerland; 8 Internal Medicine, Hôpital Neuchâtelois, 

Neuchâtel, Switzerland; 9 Gastroenterology, Hepatology and Clinical 

pathology, University Hospital, Geneva, Switzerland 

BACKGROUND: Sex specific differences in liver disease 

account for less hepatic decompensation in female cirrhotics, 

higher spontaneous HCV clearance and better chronic hepatitis 

C treatment response. Preclinical evidence suggests that 

relaxin, a pregnancy specific hormone, shows antifibrotic 

features. We aimed to evaluate the influence of past pregnancies 

on the rate of liver fibrosis progression and sustained 

virologic response (SVR) in women. METHODS: We extracted 

baseline and follow up demographic and clinical data on 

all women enrolled in Swiss Hepatitis C Cohort Study since 

2001. Differences between groups (women with/without history 

of pregnancy) were investigated using Fisher’s exact test 

for categorical variables and t-test for continuous variables. 

To reduce potential confounders, propensity score techniques 

were used. Cox proportional hazards model was used to analyze 

associations between predictors and cirrhosis taking into 

account the time to event (diagnosis of HCV infection to cirrhosis 

diagnosis). RESULTS: Among 1669 women included in 

the cohort, 1121 reported at least one pregnancy. Baseline 

characteristics were similar although women with pregnancy 

were significantly older at baseline, at time of first documented 

HCV serology and had longer follow-up time while non-pregnant 

women were more often co-infected with HIV. Overall, 

19.7% of patients were cirrhotic at a mean time from first documented 

HCV serology at 9.5 years, which was similar in both 

groups. Associated with cirrhosis were HCV therapy (HR 2.86, 

CI 2,39-3.41, p < 0.001), co-infection with HIV (HR 3.61, 

CI 2.68-4.86, p < 0.001), HCC (HR 2.47, CI 1.87-3.27, 

p < 0.001), diabetes (HR 2.50, CI 2.01-3.11, p < 0.001), 

and alcohol abuse (HR 2.08, CI 1.69-2.57, p < 0.001). Pregnancy 

was associated with a 18% risk reduction of cirrhosis 

(HR 0.82, CI 0.70-0.96, p = 0.015). Overall 39% of women 

received at least one HCV treatment with improvement or no 

change in fibrosis score. However, influence of pregnancy was 

non-conclusive. A history of pregnancy was associated with 

higher likelihood of achieving SVR with interferon-alpha-based 

antiviral therapy (OR 1.31, CI 0.95-1.81, p = 0.10). CONCLU- 

SIONS: Our data suggest that pregnancy is associated with a 

reduced risk of progression to cirrhosis as well as increased 

response to interferon-alpha- based HCV therapy. To further 

confirm a positive association between past pregnancies and 

lower rate of fibrosis progression we will integrate different 

fibrosis tests in further analyses. 

Disclosures: 





The following authors have nothing to disclose: Christine Pocha, Pascal Benkert, 

Salome A. Keller, Beat Müllhaupt, David Semela, Markus H. Heim, Darius 

Moradpour, Andreas Cerny, Raffaele Malinverni 

1885 

Safety and Efficacy of Interferon/Ribavirin-Free Therapy 

in Septuagenerians and Octogenerians with Chronic 

Hepatitis C 

Rafael Stern 1 , Karin Kozbial 1 , Clarissa Freissmuth 1 , Stephanie 

Hametner 2 , Ramona Al Zoairy 3 , Stephan Moser 4 , Asia 

Karpi 4 , Hermann Laferl 5 , Rudolf E. Stauber 6 , Heinz M. Zoller 3 , 

Andreas Maieron 2 , Wolfgang Vogel 3 , Ivo Graziadei 7 , Michael 

Gschwantler 4 , Harald Hofer 1 , Peter Ferenci 1 ; 1 Department of Internal 

Medicine III, Medical University of Vienna, Vienna, Austria; 

2 Department of Gastroenterology, Elisabethinen Hospital, Linz, 

Austria; 3 Department of Medicine II, Medical University of Innsbruck, 

Innsbruck, Austria; 4 Department of Medicine IV, Wilheminen-Hospital, 

Vienna, Austria; 5 Department of Internal Medicine, 

Kaiser-Franz-Josef-Hospital, Vienna, Austria; 6 Department of Internal 

Medicine, Medical University of Graz, Graz, Austria; 7 Department 

of Internal Medicine, Krankenhaus Hall, Hall, Austria 

Specific objective: Direct acting antivirals (DAA) have revolutionized 

treatment of chronic hepatitis C. Septua- and octogenarians 

were not included in clinical studies and therefore 

data on efficacy and safety of IFN-free treatments in the elderly 

population are missing. Elderly patients may have a decreased 

renal and hepatic function. Furthermore many are at risk for 

drug-drug interactions of DAA with drugs required for longterm 

treatment of other medical conditions. We investigated 

safety and efficacy of interferon (IFN) and ribavirin (RBV)-free 

treatments in patients >70 years with advanced liver disease 

due to chronic hepatitis C. Methods: 44 otherwise healthy 

patients >70 years (mean age: 74.6, range 70-88, m/f: 

19/26, cirrhosis: 32, F3:12, median HCV RNA 1.18x10 6 

IU/ml, HCV-genotype: GT-1a:6; 1b:34; 3:1; 4:2; missing: 

1; treatment experienced: 25; median platelet count: 137


G/l; median albumin: 39.5 g/l; median bilirubin: 0.86 mg/ 

dl; median GFR using Cockroft-Gault formula: 66.6 ml/min) 

were enrolled. The patients were treated with sofosbuvir (SOF) 

400mg/day combined either with daclatasvir (DCV) 60mg/ 

day, simeprevir (SMV) 150mg/day, or ledipasvir (LPV) 90 

mg/day. HCV RNA quantification was carried out with Abbott 

RealTime HCV (ART) assay (LLOQ


Table 1. 

CI: 95% confidence intervals; AUC: Area under Receiver Operating 

Curve; LR-likelihood ratio 

Disclosures: 

Keyur Patel - Advisory Committees or Review Panels: Merck; Consulting: Gilead 

Sciences, Santaris, Akros, Nitto Denko; Grant/Research Support: Bristol Myers 

Squibb 

The following authors have nothing to disclose: Bassem Matta, Julius M. Wilder, 

Tzu-Hao Lee 

1888 

The use of hepatitis C core antigen assay in the monitoring 

of treatment with direct antiviral agents in patients 

with chronic hepatitis C 

Elisabetta Loggi 1 , Carmela Cursaro 1 , Alessandra Scuteri 1 , Ranka 

Vukotic 1 , Martina Pirillo 1 , Marzia Margotti 1 , Valeria Guarneri 1 , 

Silvia Galli 2 , Giuliano Furlini 2 , Claudio Galli 3 , Maria Paola Landini 

2 , Pietro Andreone 1 ; 1 Medical and Surgical Sciences, University 

of Bologna, Bologna, Italy; 2 OU Microbiology and Virology, 

Azienda Ospedaliera di Bologna, Bologna, Italy; 3 Scientific 

Affairs, Abbott Diagnostics, Rome, Italy 

Background and Aim: Treatment of chronic hepatitis C (CHC) 

with direct antiviral antigens (DAAs) achieve high rates of sustained 

virological response (SVR), accompanied by few side 

effects, short duration, and a simplified route of administration. 

The implementation of these therapies have to face the problems 

of high costs and their management, that includes the efficacy 

monitoring. While HCV-RNA measured by sensitive molecular 

amplification techniques represents the gold standard, hepatitis 

C core antigen (HCV-Ag) is emerging as an interesting new 

tool in diagnosis and treatment monitoring for CHC. However, 

its role in DAAs therapies has not been still evaluated. The aim 

of our ongoing study is to evaluate the accuracy of HCV-Ag 

measurement in CHC patients treated with different schedules 

of DAAs. Methods: To date, 27 patients (21 male, median 

age 55, range 25-78) have started interferon-free (IFN-f) DAAs 

treatment for 12 or 24 weeks, and 11 completed the therapy 

course. Sero-virological testing was carried out at baseline, 

after 1-2 weeks and every 4 weeks thereafter. HCV-RNA was 

quantified by real time PCR (Amplicor, Roche; limit of detection 

15 IU/mL); HCV-Ag was assessed by an automated chemiluminescent 

two-step immunoassay (Abbott HCV core antigen 

assay; limit of detection 3 fmol/L). Results: HCV-RNA and 

HCV-Ag were highly correlated, considering both only baseline 

values (Spearman r =0.93, p


mortality. Therefore it is crucial to early make the differential 

diagnosis between the two forms in order to plan the antiviral 

treatment. 

Disclosures: 


Gilead 

The following authors have nothing to disclose: Salvatore Stefano Sciarrone, 

Alberto Zanetto, Martina Gambato, Maria Alba Diaz, Alberto Ferrarese, Massimo 

Rugge, Marco Senzolo, Giacomo Germani, Francesco P. Russo, Patricia 

Llovet, Patrizia Burra 

1890 

Evaluation of the role of hepatic progenitor cells and 

cells resistant to apoptosis in prediction of disease 

progression and treatment response in patients with 

chronic hepatitis C 

Maha El Sabaawy 2 , Eman Abdelsameea 2 , Ayat Abdallah 3 , Ahmed 

El Refaie 1 , Mervat M. Soltan 1 , Nermine Ehsan 1 ; 1 Pathology, Natioanl 

Liver Institute, Menoufia, Egypt; 2 Hepatology, Natioanl Liver 

Institute, Menoufia, Egypt; 3 Community Medicine, Natioanl Liver 

Institute, Menoufia, Egypt 

Background: Recent studies have shown increase expression 

of hepatic progenitor cells and cells resistant to apoptosis in 

chronic HCV infection. However their role in predicting disease 

progression or treatment response was not clearly elucidated. 

Aim of this study: was to investigate the role of hepatic progenitor 

cells and cells resistant to apoptosis in disease severity and 

progression along with HCV treatment response by immunohistochemistry 

(IHC). Methods: This retrospective case control 

study was conducted on 10 healthy control subjects (donors 

for liver transplantation) and 91 chronic HCV patients who 

had completed combination treatment (interferon plus ribavirin). 

Demographic and clinical characteristics were collected 

from the data registry. Patients were categorized according 

to their SVR (sustained virological response) into two groups; 

responders and non-responders. Paraffin blocks from liver 

biopsies of control and patients included in this study were 

retrieved. Semi-thin sections were prepared on positive charge 

slides for immunostaining with CK7, Ki67 and bcl2 antibodies. 

Results: No specific age or sex prediction was reported 

in patients’ study group concerning any of the investigated 

markers. Control healthy subjects showed negative immunoreaction 

to hepatic progenitor cells either isolated or ductular, 

Ki67 and bcl2 lymphocyte associated portal tracts (LPT). One 

case of control group that revealed 15% steatosis had positive 

immunoreaction to bcl2 lymphocyte associated hepatic parenchyma 

(LAH). Laboratory data including transaminases (AST 

and ALT), platelets and prothrombin time exhibited significant 

relation with Ki67, CK7 both isolated and ductular and bcl2 

both LPT and LAH. CK7 ductular showed significant association 

with fibrosis and necroinflammatory activity according to Ishak 

score (P< 0.05) while non-significant relation was noticed in 

the CK7 isolated form and Ki67 (P> 0.05). Moreover bcl2 

both (LPT) and (LAH) demonstrated significant association with 

fibrosis and necroinflammatory activity (P< 0.05). Pertaining 

to SVR significant relation was confirmed with Ki67, CK7 both 

isolated and ductular and bcl2 (PT) and (LAH) (P< 0.05). Multivariate 

analysis revealed their role as predictors of HCV treatment 

response. Conclusion: Hepatic progenitor cells and cells 

resistant to apoptosis are significantly related to disease progression 

and could predict response of combination treatment 

in chronic HCV. 

Disclosures: 

The following authors have nothing to disclose: Maha El Sabaawy, Eman Abdelsameea, 

Ayat Abdallah, Ahmed El Refaie, Mervat M. Soltan, Nermine Ehsan 

1891 

Cascade of Care of HCV & HIV Infected Patients Identified 

Through Community Pop-Up Clinics (CPCs) 

Syune Hakobyan; Vancouver ID Research and Care Centre Society, 

Vancouver, BC, Canada 

AUTHORS: S. Hakobyan, S. Sharma, A. King, F. Zahedieh, 

H. Tossonian, B. Conway. BACKGROUND: The prevalence 

of Hepatitis C Virus (HCV) and Human Immunodeficiency 

Virus (HIV) infections is very high among the 18,000 men and 

women of Vancouver’s Downtown East Side (DTES). Despite 

the widespread availability of medical and non-medical services, 

there is little information regarding the cascade of care 

for those diagnosed with HIV and/or HCV in this neighborhood. 

The aim of this study was to document the cascade of 

HIV and HCV care in the DTES and identify obstacles to its 

improvement. METHODS: Participants were recruited at CPCs 

held at different community-based centers in the DTES frequented 

by people who inject drugs (PWID). During the CPCs, 

OraQuick HIV and HCV Rapid Antibody point of care testing 

was offered and participants were then asked to complete a 

targeted questionnaire while they waited for test results. Care 

for HIV immediately offered to those who were infected, while 

the questionnaire answers were used to identify parameters of 

engagement around HCV infection. RESULTS: From 03/13 to 

05/15, 1410 individuals were tested for HIV and HCV. Of 

these, 448 (31.8%) were found to be infected with HCV and 

29 (2.1%) co-infected with HIV. Within a linked multidisciplinary 

clinic, 18 HIV-infected individuals (62.1%) were linked 

to care with 9 (50%) achieving an undetectable viral load on 

treatment. In evaluating the feasibility of expanding the program 

to address HCV infection, 910 participants completed a 

questionnaire (22.9% female, 56.5% Caucasian, 28.5% First 

Nations; mean age: 46.3). Amongst all survey participants, 24 

were co-infected with HIV and HCV. In this population, 41.7% 

were Aboriginal, 75.0% had injected drugs, 29.2% shared 

needles and other injection equipment, 62.5% were previously 

incarcerated, and 75.0% were aware of a HCV cure, and 

87.5% stated they would consider treatment if they had HCV, 

although 54.2% did not believe they needed it. CONCLU- 

SIONS: Despite extensive prior testing for HIV on the DTES, we 

identified 29 individuals who were unaware or unengaged in 

care, and could arguably be considered as “core transmitters” 

of HIV and HCV. Their engagement in treatment for HIV has 

been successful in our model of care. There is significant interest 

in considering HCV treatment were it offered. CPCs are an 

important tool in initially engaging these individuals in medical 

care and moving towards increased HCV treatment uptake in 

this population. 

Disclosures: 

The following authors have nothing to disclose: Syune Hakobyan 

1892 

Correlates of Successful HCV Treatment in HIV Co-Infected 

Vulnerable Populations 

Brian Conway; Vancouver ID Research and Care Centre Society, 

Vancouver, BC, Canada 

AUTHORS: B. Conway, S. Hakobyan, A. King, F. Zahedieh, 

H. Tossonian, S. Sharma INTRODUCTION: Vulnerable populations, 

including people who inject drugs (PWID), are over-represented 

among HIV/HCV co-infected populations. Although 

clinical trial results suggest that many treatment regimens for 

HCV infection are equally effective in the setting of HIV co-infection, 

this has not been clearly established in populations 

consisting mainly of PWID and members of related vulnerable


inner city groups. METHODS: We have established a multi-disciplinary 

outreach program to recruit and retain HIV/HCV-infected 

PWID in care. The program includes access to specialty 

medical care, extended social support services, complimentary 

over-the-counter medications and vitamins, daily snacks/beverages, 

and weekly meals and HIV/HCV support groups. HIV 

and HCV treatment is offered to all who qualify for it on medical 

grounds. We have conducted a retrospective analysis of 

all HIV co-infected patients treated for HCV infection within our 

program and for whom a definite HCV treatment outcome was 

ascertained. This analysis correlates SVR with a range of baseline 

demographic and clinical variables, including housing and 

active recreational drug use. RESULTS: Of 512 HIV-infected 

individuals in our program, 245 (47.9%) are co-infected with 

HCV, including 172 (70.2%) active (current/recent) PWID. 

In total, 75 (30.6%) have completed HCV treatment (5 on alloral 

regimens), including 55 (73.3%) active PWID, and 46 

(61.3%) with genotype 1 infection. The mean age was 51, 70 

(93.3%) male, 22 (26.7%) on opiate substitution, 72 (96.0%) 

on HIV treatment (62/72 with full virologic suppression), 20 

(27.8%) homeless, and 32 (42.7%) attending weekly HCV 

support groups. The SVR rate was 43.1% (31/72), 80% (4/5) 

in genotype 1 patients on all-oral regimens. SVR was higher 

in non-genotype 1 (54.5% vs. 43.1%) patients. Success rates 

were no higher in subjects on methadone 12 (54.5%), and no 

lower in the homeless 10 (50.0%) or active PWID 25 (45.5%). 

SVR was over 56% in active support group participants. An 

additional 9 co-infected patients have been started on all-oral 

regimens, and SVR data will be presented. DISCUSSION: In our 

centre, >35% of HIV/HCV co-infected patients (mostly PWID) 

have been treated for HCV infection. Many of the usual negative 

predictive factors do not affect treatment success, a fact 

that may be linked to the low-barrier multi-disciplinary nature 

of our program. There is a trend towards increased success in 

patients more actively engaged in our program. Our model will 

provide an opportunity for vulnerable populations to benefit 

from treatment for both HIV and HCV infections. 

Disclosures: 





time intervals until postoperative recurrence (within 2 years, 

after 2 years or later, and no recurrence). (1) The maximum 

standardized uptake value (SUV max) of tumor was assessed 

and the best cut-off value of SUV max for predicting MVI was 

obtained using receiver operating characteristic (ROC) curve. 

Independent predictors for MVI were identified by multivariate 

analysis. (2) Using the best cut-off value of SUV max for predicting 

MVI, the associations between SUV max and recurrence 

patterns and intervals were analyzed. Results: (1) 31 of 79 

(39.2%) patients showed positive preoperative PET-scans and 

the mean SUV max was 3.15 ± 1.28 (range, 2−8.8). MVI 

was present in 13 of 79 patients (16.5%) on pathologic examination. 

SUV max ≥5.0 was suggested to be the best cut-off 

value for predicting MVI by ROC curve (area under the curve 

[AUC] =0.712, sensitivity=46.1%, specificity=90.6%). Multivariate 

analysis showed that SUV max ≥5.0 (p=0.05), AFP- 

L3 ≥15% (p=0.02), and non-simple nodular as macroscopic 

classification (p=0.039) were independent predictors for MVI. 

(2) SUV max in the recurrence within the MC (3.25±0.94) 

and no recurrence group (2.92±0.86) was significantly lower 

than that in the recurrence beyond the MC group (4.98±2.79). 

Multivariate analysis showed that SUV max ≥5.0 (p=0.002) 

was an independent predictor for recurrence beyond the MC. 

SUV max in the group with recurrence after 2 years or later 

(3.45±1.56) and no recurrence (2.92±0.86) was significantly 

lower than that in the within 2 years group (4.06±2.14). Multivariate 

analysis showed that SUV max ≥5.0 (p=0.002) and 

tumor size ≥30mm (p=0.026) were independent predictors for 

recurrence within 2 years. Conclusions: Preoperative 18 F-FDG 

PET-CT predicts MVI, recurrence beyond the MC, and recurrence 

within 2 years after curative HR for early-stage HCC. 

Disclosures: 








The following authors have nothing to disclose: Tomoki Kobayashi, Hiroshi 

Aikata, Fumi Shinohara, Norihito Nakano, Yuki Nakamura, Masahiro Hatooka, 

Kei Morio, Reona Morio, Takayuki Fukuhara, Yuuko Nagaoki, Tomokazu Kawaoka, 

Akira Hiramatsu, Michio Imamura, Yoshiiku Kawakami 

1893 

High 18 F-FDG uptake on preoperative PET-CT correlates 

with microvascular invasion and predicts early and 

massive recurrence after curative resection for early-stage 


Tomoki Kobayashi, Hiroshi Aikata, Fumi Shinohara, Norihito 

Nakano, Yuki Nakamura, Masahiro Hatooka, Kei Morio, Reona 

Morio, Takayuki Fukuhara, Yuuko Nagaoki, Tomokazu Kawaoka, 

Akira Hiramatsu, Michio Imamura, Yoshiiku Kawakami, Kazuaki 

Chayama; Department of Gastroenterology and Metabolism, Hiroshima 

University Hospital, Hiroshima, Japan 

BackgroundAim: Microvascular invasion (MVI) predicts recurrence 

beyond the Milan criteria (MC) and early recurrence 

after hepatic resection (HR) for hepatocellular carcinoma 

(HCC). Recently, several studies have shown that 18 F-FDG 

uptake may be a surrogate marker for MVI. This study aimed to 

assess the value of preoperative 18 F-FDG PET-CT for predicting 

recurrence patterns and intervals after HR in early-stage HCC. 

Methods: 79 consecutive patients who were preoperatively 

examined with 18 F-FDG PET-CT and underwent curative HR for 

HCC within the MC were enrolled in this retrospective cohort 

study. They were classied according to the recurrence patterns 

(beyond the MC and within the MC or no recurrence) and the 

1894 

Reduced sensitivity of ultrasound in detecting hepatocellular 

carcinoma in obese patients, using explants 

pathology as gold standard 

Jamak Modaresi Esfeh 2 , Kaveh Hajifathalian 1 , Kianoush Ansari-Gilani 

3 , Ahyoung Kim 1 , Carlos J. Romero-Marrero 2 ; 1 Internal Medicine, 

Cleveland clinic, Beachwood, OH; 2 Gastroenterology and 

Hepatology, Cleveland Clinic, Cleveland, OH; 3 Diagnostic Radiology, 

University Hospitals, Case Medical Center, Cleveland, OH 

Background: American Association for the Study of Liver 

Diseases (AASLD) recommends screening for hepatocellular 

carcinoma (HCC) among patients with cirrhosis to be done 

with ultrasound (US) every six months, regardless of their BMI. 

We examined the sensitivity of US for diagnosis of HCC in 

obese patients. Methods: We used the prospective data from 

liver transplant patients between January 2005 and December 

2010, who were diagnosed with HCC in the pathologic examination 

of their explants. All patients had US studies of their 

liver within six months of diagnosis of HCC. Number and size 

of HCC lesions were extracted from radiologic and pathologic 

reports. Obesity was defined based on BMI≥30 kg/m 2 . Results: 

99 patients, 22% female, with mean BMI of 31 kg/m 2 (range 

20-43) were included. Most common underlying liver disease


was HCV (52%). At diagnosis, median number of HCC lesions 

was 1 (IQR 1-2) and median size of the largest lesion was 2 cm 

(IQR 1.5-3.1). Overall sensitivity of an US study for detection of 

HCC was 38% (95% CI 29-48%). Sensitivity was 77% (95% CI 

62-93%) in patients with BMI


1897 

Diagnostic Performance of Contrast-enhanced Imaging 

Techniques for Small Hepatocellular Carcinoma Measuring 

less than 2 cm 

Seong Hee Kang 1 , Jong Eun Yeon 1 , Young-Sun Lee 1 , Tae Suk 

Kim 1 , Yang Jae Yoo 1 , Sang Jun Suh 1 , Young Kul Jung 1 , Ji Hoon 

Kim 1 , Hyung Joon Yim 1 , Chang Hee Lee 2 , Kwan Soo Byun 1 ; 1 Gastroenterology 

& Hepatology, Korea University Medical Center, 

Seoul, Korea (the Republic of); 2 Radiology, Korea University Medical 

Center, Seoul, Korea (the Republic of) 

BACKGROUND: Recent guidelines recommend ultrasound follow-up 

for nodules (< 1 cm) discovered via ultrasonography 

surveillance (US) in liver cirrhosis patients. Nodules larger than 

1cm should be followed the recall policy with either dynamic 

contrast 4-phase CT scan or dynamic enhanced contrast MRI. 

There is no specified recall policy of nodule less than 1 cm 

found on US. We evaluated CT and MRI efficacy in hepatocellular 

carcinoma (HCC) diagnosis in nodules lesser than 

1cm and 1~2 cm. METHODS: We retrospectively analyzed 

130 ultrasonography-detected liver nodules


1899 

Efficacy of Delta-AFP in Predicting the Development of 

Hepatocellular Carcinoma in Cirrhotic Patients 

Ross Mund 1 , Yanhong Deng 2 , Maria Ciarleglio 2 , Tamar H. Taddei 

1,3 ; 1 Yale University School Of Medicine, New Haven, CT; 

2 Yale Center for Analytical Sciences, New Haven, CT; 3 VA Connecticut 

Healthcare System, West Haven, CT 

Background: Current AASLD practice guidelines recommend 

abdominal ultrasound every six months for hepatocellular carcinoma 

(HCC) screening in at risk individuals. Serum AFP was 

removed from guidelines in 2011 due to poor sensitivity. This 

study evaluated the change in AFP over time (delta AFP) as 

a predictor of HCC development in cirrhotic patients. Methodology: 

This retrospective cohort study included all cirrhotic 

patients without HCC from the VA Connecticut Healthcare System 

who had a baseline AFP and two subsequent AFP measurements 

during 2006-2011. AFP was measured at baseline, first 

serial AFP at an average of 320.88 days (std dev 103.13), 

and last serial AFP at an average of 605.75 days (std dev 

122.21) from the baseline. The absence of HCC was defined 

by absence of a focal hepatic mass consistent with HCC on 

dynamic imaging from the time of baseline AFP to the last measurement. 

Subjects were classified according to whether they 

had hepatitis C (HCV) cirrhosis or not. Longitudinal analysis 

with patient level random effects were used to model change 

in AFP between those who developed and did not develop 

HCC. Logistic regression analysis was used to examine the link 

between delta AFP and HCC. Results: 169 subjects met criteria 

for the study. N=91 for HCV cirrhosis (53.85%). 17 subjects 

developed HCC (10.06%). Delta AFP was assessed between 

the two groups. There was significant difference in delta AFP 

from baseline to the first serial measurement between HCC-N 

and HCC-Y groups (+0.2 and +8.75, respectively, p=0.0362, 

95% CI: 0.56-16.55), and from the baseline to the last AFP 

measurement (+1.72 and +85.73, respectively, p +20 were 5.342 times more likely 

to develop HCC as compared to those with delta AFP < +20 

(p= 0.0067; 95% CI: 1.593-17.919). HCV as a moderator 

could not be assessed due to limited sample size. Conclusion: 

This study revealed statistically significant differences in delta 

AFP in cirrhotic subjects who developed HCC versus those who 

did not develop HCC. Importantly, this difference in delta AFP 

was seen at the first serial AFP value, an average of 284.87 

days prior to the development of HCC. Additionally, the risk of 

developing HCC for subjects with a delta AFP > +20 over a 

2 year span was significantly increased. Due to small sample 

size and wide confidence intervals, this study should be validated 

in a larger, prospective cohort. This study supports the 

hypothesis that AFP may be a cost effective, readily available 

and early biomarker for the diagnosis of HCC, once we better 

understand thresholds and changes that may increase its sensitivity. 

Disclosures: 

Tamar H. Taddei - Consulting: Onyx Pharmaceuticals; Grant/Research Support: 

Bayer HealthCare 

The following authors have nothing to disclose: Ross Mund, Yanhong Deng, 

Maria Ciarleglio 

1900 

A mass spectrometry based serum test for the detection 

of hepatocellular carcinoma (HCC) in high risk patients 

Devalingam Mahalingam 1 , Julio A. Gutierrez 1 , W. Kenneth Washburn 

1 , Glenn A. Halff 1 , Leonidas Chelis 2 , Stylianos Kakolyris 2 , 

Stylianos Vradelis 3 , Julia Grigorieva 4 , Carlos Oliveira 4 , Heinrich 

Roder 4 , Joanna Roder 4 ; 1 Cancer Therapy and Research Center, 

University of Texas Health Sciences Center San Antonio, San Antonio, 

TX; 2 Department of Medical Oncology, Democritus University 

of Thrace, Alexandroupolis, Greece; 3 2nd Department of Internal 

Medicine, Democritus University of Thrace, Alexandroupolis, 

Greece; 4 Biodesix, Boulder, CO 

Improved screening protocols (SP) for patients at high risk of 

developing HCC could lead to improved patient outcome and 

possibly cure if detected early. The current SPs, ultrasound with 

the possible addition of alpha-fetoprotein (AFP) measurement, 

suffer from insufficient sensitivity and specificity, and less than 

30% of patients are diagnosed early enough to be suitable 

candidates for resection or transplantation. A better biomarker 

for the early detection of HCC is clearly needed. We used 

mass spectrometry analysis of serum samples combined with 

specialized deep learning techniques to train and validate such 

a test. Samples were available from two patient cohorts. The 

first was of patients undergoing transplant or resection for HCC 

(N=48; median MELD = 14), and patients with advanced cirrhosis 

undergoing liver transplant (N =52; median MELD = 

25); underlying cause of liver disease was 49% hepatitis C 

infection (HCV). The second cohort consisted of patients with 

advanced HCC (N= 110; 72/27/11 Child-Pugh A/B/C) and 

liver disease but no HCC (N= 83; 74/7/2 Child-Pugh A/B/C), 

with 68% of patients having hepatitis B infection (HBV). The 

two cohorts were combined to create a more representative set 

of patients with and at risk of HCC, which was split into development 

and validation sets, stratified by cause and severity 

of liver disease and stage of HCC. The classifier was trained 

especially to avoid confounding by liver function. The resulting 

test showed a sensitivity/specificity of 83%/84% in cross 

validation in the development set, with 73% of patients with 

HCC who could be treated by resection or transplant (BCLC 

A) correctly identified. Performance was good across all represented 

causes of liver disease. Validation set performance was 

similar with sensitivity/specificity of 81%/79%, an accuracy of 

64% for patients with BCLC A HCC, and little dependence on 

cause of liver disease (overall accuracy: 91% HBV, 79% HCV). 

Within the validation set, when compared with AFP alone, 

the test had 22% greater specificity at fixed sensitivity and 

12% greater sensitivity at fixed specificity. Results from further 

ongoing validation of the test on independent patient cohorts 

will also be presented. The developed test shows promise in 

the early detection of HCC as a screening tool in high risk 

patients independent of the cause of underlying liver disease. 

While further validation will be necessary to conclusively prove 

its clinical validity, we believe such a test could substantially 

improve early detection of HCC. 

Disclosures: 

Glenn A. Halff - Board Membership: texas organ sharing alliance, texas organ 

sharing alliance, texas organ sharing alliance, texas organ sharing alliance; 

Stock Shareholder: xenex, xenex, xenex, xenex 

Julia Grigorieva - Stock Shareholder: Biodesix 

Carlos Oliveira - Employment: Biodesix; Patent Held/Filed: Biodesix 

Heinrich Roder - Employment: Biodesix 

Joanna Roder - Employment: Biodesix; Patent Held/Filed: Biodesix; Stock Shareholder: 

Biodesix 

The following authors have nothing to disclose: Devalingam Mahalingam, Julio 

A. Gutierrez, W. Kenneth Washburn, Leonidas Chelis, Stylianos Kakolyris, Stylianos 

Vradelis


1901 

ARID1A mutations may represent a novel pathway of 

carcinogenesis in biliary carcinomas 

Motoko Sasaki 1 , Takeo Nitta 2 , Yasunori Sato 1 , Yasuni 

Nakanuma 3 ; 1 Human Pathology, Kanazawa University Graduate 

School of Medicine, Kanazawa, Japan; 2 Gastroenterological 

Surgery II, Hokkaido University Graduate School of Medicine, 

Sapporo, Japan; 3 Division of Pathology, Shizuoka Cancer Center, 

Shizuoka, Japan 

Backgrounds/Aims. We have reported that some cholangiocarcinoma 

arise following a stepwise pathway of carcinogenesis 

through flat and papillary premalignant lesions, biliary intraepithelial 

neoplasia (BilIN) and intraductal papillary neoplasm of 

the bile duct (IPNB) and KRAS mutations and GNAS mutations 

may be involved in this pathway. Recent studies using exome 

sequencing and next generation sequencing revealed frequent 

alterations in several new genes including the three chromatin-remodeling 

genes (BAP1, ARID1A or PBRM) in cholangiocarcinomas. 

Given frequent inactivating mutations in ARID1A 

gene coding BAF250a protein in intrahepatic cholangiocarcinoma 

and a reported usefulness of ARID1A/BAF250a immunostaining 

to detect ARID1A mutation, this study investigates 

immunohistochemically the clinicopathological significance of 

ARID1A mutations in biliary carcinomas. Methods. We examined 

the status of inactivating mutations in ARID1A by immunohistochemistry 

and the relationship with clinicopathological 

features in 13 patients with combined hepatocellular-cholangiocarcinoma 

(cHC-CC, M/F=9/4), 49 patients with intrahepatic 

cholangiocarcinoma (ICC, M/F=22/27), 17 with IPNB (M/ 

F=11/6), 72 with extrahepatic cholangiocarcinoma (EHCC, 

M/F=51/21) and 43 with gallbladder carcinoma (GBC, M/ 

F=18/25). Fifteen BilIN lesions were also examined. Results. 

Complete loss or clonal loss of ARID1A immunoreactivity indicating 

the inactivating mutations of ARID1A was detected in 

one (7.7%) of cHC-CC, 9 (18.4%) of ICC, none of IPNB, 11 

(15.3%) of EHCC and 4 (9.1%) of GBC. The patients with 

biliary carcinoma with ARID1A mutations were significantly 

female-predominant (p


cirrhosis etiology in cirrhotic patients undergoing screening 

for HCC. Methods: This was a case-control study conducted 

on patients with cirrhosis with and without HCC identified via 

ICD-9 diagnosis codes retrospectively from 1990-2014 in a 

large state-wide database who underwent surveillance imaging. 

HCC was diagnosed by imaging and/or biopsy. The cirrhosis 

cohort without HCC was determined by absence of HCC 

on last imaging study. AFP levels within 90 days of imaging 

were abstracted. The sensitivity and specificity of AFP levels 

was determined for HCC diagnosis in HCV cirrhosis, non-viral 

cirrhosis and the entire cohort. AFP thresholds were explored 

based on previous literature by ROC analysis and group comparisons 

were performed by Chi-square tests. Results: 10,481 

patients with cirrhosis were identified, 6166 patients excluded 

(no AFP levels, no AFP levels within 90 days, or AFP >500). 

1641 patients with HCC were identified, 402 excluded as 

HCC diagnosed before first imaging study. 3371 patients with 

cirrhosis and 551 HCC patients were analyzed. Patients were 

predominantly white (87 % without HCC, 86 % with HCC), 

and male (59% cirrhotic cohort, 72% HCC cohort). Patients 

with cirrhosis were younger (54 vs 57 years p


1905 

WITHDRAWN 

1906 

LI-RADS hepatocellular carcinoma diagnostic classification 

system: utilization by community radiologists, and 

results of second opinion reading by staff radiologists at 

a transplant center 

Jesse M. Civan 1 , Aaron Martin 2 , Raza Hasan 2 , Flavius Guglielmo 

3 , Sandeep Deshmukh 3 , Christopher G. Roth 3 , Donald G. 

Mitchell 3 ; 1 Gastroenterology & Hepatology, Thomas Jefferson University 

Hospital, Philadelphia, PA; 2 Internal Medicine, Thomas Jefferson 

University, Philadelphia, PA; 3 Radiology, Thomas Jefferson 

University, Philadelphia, PA 

Background: LI-RADS (Liver Imaging Reporting and Data System) 

was created to clarify radiographic degree of concern 

for hepatocellular carcinoma (HCC). In 2013, UNOS adopted 

criteria analogous to LI-RADS category 5 (definite HCC) for 

lesions to qualify for HCC MELD exception. We evaluated 

LI-RADS utilization by community radiologists, inter-operator 

agreement of LI-RADS scoring, and prediction of LI-RADS classification 

by descriptive language. Methods: We reviewed 150 

outside abdominal MRI and CT studies that were overread 

(formal consultation) by radiology specialists at a transplant 

center. Studies performed at other academic centers were 

excluded. We abstracted qualitative phrases used by outside 

radiologists in lieu of LI-RADS to convey degree of concern 

for HCC. Statistical analysis was performed using SPSS v23. 

Results: LI-RADS classification was provided in 13% of cases by 

outside radiologists and 88% of internal re-reads. For outside 

reports utilizing LI-RADS, correlation with our LI-RADS classification 

was poor, Spearman 0.00 (p=0.99) for combined 

LI-RADS 4/5, Spearman 0.07 (p=0.82) for LI-RADS 5. For outside 

reports not utilizing LI-RADS, 16 distinct qualitative phrases 

were used to convey degree of concern for HCC, many of 

which did not map clearly to a LI-RADS category (e.g. “in 

keeping with HCC”, “could represent HCC”). Overall, descriptive 

terms did not predict a combined outcome of the presence 

of LI-RADS 4 or 5 lesions (Cramer’s V=0.48, p=0.27), 

or presence of LI-RADS 5 lesions alone (Cramer’s V=0.56, 

p=0.06). Correlation between individual descriptive phrases 

and LI-RADS classification is depicted in figure 1. Conclusion: 

The LI-RADS classification system is under-utilized in the community 

setting. Qualitative language used in outside radiology 

reports poorly predicted diagnosis of probable or definite HCC 

by LI-RADS criteria. 

Disclosures: 


The following authors have nothing to disclose: Aaron Martin, Raza Hasan, 

Flavius Guglielmo, Sandeep Deshmukh, Christopher G. Roth, Donald G. Mitchell 

1907 

A Model to Estimate Survival in Ambulatory Patients 

with Hepatocellular Carcinoma: Can It Predict the Natural 

Course of Hepatocellular Carcinoma? 

Won-Mook Choi 1 , Su Jong Yu 2 , Young Youn Cho 2 , Eun Ju Cho 2 , 

Jeong-Hoon Lee 2 , Yoon Jun Kim 2 , Jung-Hwan Yoon 2 , June Sung 

Lee 3 ; 1 Graduate School of Medical Science and Engineering, 

Korea Advanced Institute of Science and Technology(KAIST), Daejeon, 

Korea (the Republic of); 2 Department of Internal Medicine 

and Liver Research Institute, Seoul National University College of 


Medicine, Ilsan Paik Hospital, Inje University College of Medicine, 

Goyang, Korea (the Republic of) 

Objectives: Several hepatocellular carcinoma (HCC) staging 

systems are available; however, whether these staging systems 

could predict the natural course of HCC is largely unknown. 

This study aimed to investigate whether the various HCC staging 

systems could reflect the natural course of HCC. Methods: 

1116 patients with history of HCC treatment and 111 patients 

without any history of treatment till death or last follow-up at a 

single tertiary hospital were included. To minimize selection 

bias, patients with treatment were matched using propensity-score 

matching at a 1:1 ratio with patients without treatment. 

The model’s performance was assessed and compared to other 

staging systems using C-statistics and Akaike information criterion 

(AIC). Results: In the group of treated patients before 

propensity score matching, the Model to Estimate Survival 

in Ambulatory HCC patients (MESIAH) score showed higher 

predictive performance, with a C-statistic of 0.832 (95% confidence 

interval [CI], 0.808-0.856), when compared to the 

Barcelona Clinic Liver Cancer staging system and the seventh 

edition of the American Joint Committee on Cancer TNM staging 

system. However, in the group of untreated patients, all 

staging systems including the MESIAH score failed to predict 

survival. After propensity score matching, although the MESIAH 

score was balanced between the two groups (5.89±1.35 and 

5.94±1.45, P=0.60), the treated group survived longer than 

the untreated group, which suggests that treatment by itself did 

prolong survival. Moreover, even the MESIAH score failed to 

predict outcome of not only the untreated group but also the 

treated group after propensity score matching. Conclusions: 

Although the MESIAH score provided better prognostic stratification 

than other staging systems, it also was not helpful 

in predicting the natural course of HCC. Since the preferred 

treatment modality is varied by multiple factors, it is necessary 

to develop a new staging system that reflects the natural course 

of HCC regardless of treatment.


Fig 1. Observed and predicted overall survival curves stratified 

by the MESIAH score (A) with treated groups, (B) with untreated 

groups before propensity score matching 

seem to be a good option for the subset of patients with symptomatic 

severe portal hypertension. 

Disclosures: 

The following authors have nothing to disclose: Vincent Soriano, Pablo Labarga, 

Jose V. Fernandez-Montero, Carmen de Mendoza, Pablo Barreiro 

Disclosures: 






The following authors have nothing to disclose: Won-Mook Choi, Su Jong Yu, 

Young Youn Cho, Eun Ju Cho, Jeong-Hoon Lee, Jung-Hwan Yoon, June Sung Lee 

1908 

Long-term Outcome of HIV+ Patients with Non-Cirrhotic 

Portal Hypertension Upon Discontinuation of Didanosine 

Vincent Soriano 1 , Pablo Labarga 4 , Jose V. Fernandez-Montero 2 , 

Carmen de Mendoza 3 , Pablo Barreiro 1 ; 1 La Paz University Hospital 

& IdiPAZ, Madrid, Spain; 2 University Hospital Crosshouse, 

Kilmarnock, United Kingdom; 3 Puerta de Hierro Research Institute, 

Majadahonda, Spain; 4 La Luz Clinic, Madrid, Spain 

Background: Exposure to the antiretroviral drug didanosine has 

been associated to hepatic vascular damage and non-cirrhotic 

portal hypertension (NCPH) in HIV+ individuals. Life-threatening 

episodes of variceal bleeding and acute portal thrombosis 

are the most feared complications. Herein, we describe the 

long-term outcome of a series of patients that developed HIV-associated 

NCPH. Methods: All HIV+ patients followed at one 

single HIV reference large clinic in Madrid were examined. 

Diagnosis of NCPH was made based on recognition of portal 

hypertension in the absence of cirrhosis (confirmed either by 

biopsy and/or elastometry), and exclusion of any known etiology 

other than didanosine exposure. Following removal of 

the drug, patients have been on regular follow-up using other 

antiretroviral drugs. Results: From a total of 3,200 HIV+ individuals 

attended during the last decade, a total of 21 (0.6%) 

were diagnosed with didanosine-associated NCPH. Characteristics 

features of small portal venulopathy were found in the 

liver biopsy of all 12 patients with histologically available specimens. 

At diagnosis, mean age was 43-years and 17 were 

male. No deaths due to liver-related deaths have occurred 

after an average follow-up of 8.3 years. However, two thirds 

developed serious liver-related complications early on, including 

portal thrombosis (7), ascites (8), variceal bleeding (6) and 

encephalopathy (2). Two subjects underwent TIPPS and one 

surgical porto-cava shunt due to severe portal hypertension. 

Interestingly, none of these 3 patients developed encephalopathy 

thereafter, most likely due to their relative well-preserved 

hepatic function. No further liver-related complications occurred 

beyond 2 years of didanosine removal in our series except for 

one patient that suffered variceal re-bleeding 7 years later. 

Conclusions: NCPH in HIV+ patients is a rare but potentially 

life-threatening condition linked to didanosine exposure that 

tends to ameliorate over time after stopping didanosine. TIPPS 

1909 

AMPK activation prevents and reverses drug-induced 

mitochondrial damage in hepatocytes by regulating 

mitochondrial quality control 

Dong Fu 1 , Ghada Haydar 1 , Sun Woo Sophie Kang 1 , Geoffrey C. 

Farrell 2 , Jennifer Lippincott-Schwartz 3 , Irwin M. Arias 3 ; 1 Faculty 

of Pharmacy, The University of Sydney, Sydney, NSW, Australia; 

2 Liver Research Group, National University of Australia Medical 

School, Canberra, ACT, Australia; 3 NICHD, National Institutes of 

Health, Bethesda, MD 

Mitochondrial damage plays a central role in drug-induced 

liver injury (DILI) which is responsible for many cases of acute 

liver failure, and pre- or post-market withdrawal of drugs. Mitochondria 

maintain function through quality control that includes 

regulated fusion/fission dynamics and autophagy-mediated 

degradation that eliminates damaged mitochondria. Therefore, 

enhancement of mitochondrial quality control is a potential 

approach for treatment of DILI. AMP activated kinase (AMPK) 

is a cellular energy sensor which, when activated by phosphorylation, 

promotes mitochondrial biogenesis and activates 

autophagy. Using collagen sandwich cultures of rat and human 

hepatocytes, we investigated the role of AMPK in mitochondrial 

quality control, and prevention and reversal of drug-induced 

mitochondrial and hepatocellular damage. Results: Western 

blot and immunofluorescence results demonstrated that hepatotoxic 

drugs, acetaminophen (for intrinsic DILI) and diclofenac 

(for idiosyncratic DILI), significantly decreased expression 

of mitochondrial fusion protein Mfn1; acetaminophen also 

decreased expression of fusion proteins Mfn2 and Opa1. Both 

drugs caused mitochondrial fragmentation, and decreased ATP 

levels and mitochondrial membrane potential in human and 

rat hepatocytes, revealing that both drugs cause mitochondrial 

damage. Moreover, both drugs significantly decreased cell viability 

and caused depolarization in human and rat hepatocytes. 

Activation of AMPK, by simultaneous administration of hepatotoxic 

drugs and AICAR, a specific AMPK activator, or addition 

of AICAR at 5hr post exposure of both drugs when hepatocellular 

damage occurred, restored mitochondrial function and 

fusion, hepatocyte polarization and cell viability, revealing 

that activation of AMPK prevents and reverses drug-induced 

mitochondrial and hepatocellular damage. AMPK activation 

prevented drug-mediated decreases in Mfn1, 2 and Opa1, 

indicating AMPK induced mitochondrial fusion by regulating 

fusion machinery. AMPK activation also increased autophagy 

in the presence of acetaminophen but not diclofenac, suggesting 

that the autophagic effect of AMPK plays a differential role 

in prevention of drug-induced hepatotoxicity. Conclusion: Acetaminophen 

and diclofenac fragment mitochondria through inhibition 

of the fusion machinery, and cause mitochondrial and 

hepatocellular damage. Through promotion of mitochondrial 

fusion or together with activation of autophagy, two important 

processes of mitochondrial quality control, activation of AMPK 

prevents and reverses drug-induced mitochondrial and hepatocellular 

damage. Activation of AMPK may provide a novel 

strategy for treatment of DILI. 

Disclosures: 

The following authors have nothing to disclose: Dong Fu, Ghada Haydar, Sun 

Woo Sophie Kang, Geoffrey C. Farrell, Jennifer Lippincott-Schwartz, Irwin M. 

Arias


1910 

The role of CD36 in acetaminophen-induced hepatotoxicity 

in mice 

chen zhang 1 , Xianmin Mu 1 , Che Xu 1 , Shi Hu 1 , Yuanfang Lu 1 , 

Qiang You 1,2 ; 1 Department of Biotherapy, Second Affiliated Hospital, 

Nanjing Medical University, Nanjing, China; 2 Department of 

Immunology, Nanjing Medical University, Nanjing, China 

Acetaminophen (APAP) overdose cause hepatotoxicity, and 

even acute liver failure, which involves a series of critical events 

including APAP metabolite protein adduct formation, mitochondrial 

dysfunction, oxidant stress, peroxynitrite formation and 

nuclear DNA fragmentation. Recent studies indicate that sterile 

inflammation and innate immune cells may play important roles 

in APAP-induced liver injury and repair. CD36 is a member of 

the class B scavenger receptor family of cell surface proteins. It 

binds a variety of ligands including collagen, thrombospondin, 

erythrocytes parasitized with Plasmodium falciparum, oxidized 

low density lipoprotein, native lipoproteins, oxidized phospholipids, 

and long-chain fatty acids. CD36 also assembles 

with Toll-like receptor 4 and 6 to promote sterile inflammation. 

The present study aims to investigate the role of CD36 

in APAP induced murine liver injury. Methods: WT C57BL/6J 

and CD36 -/- mice on same background were intraperitoneally 

injected with APAP (300mg/kg) after fasting for 16h. The 

blood and liver tissues were collected at 8h and 24h after 

treatment. Liver injury was evaluated by serum ALT level and 

liver tissue H&E staining. Liver inflammatory factors expression 

was determined by real time PCR. The metabolite of APAP, 

N-acetyl-p-benzoquinone imine (NAPQI) protein adducts, and 

Cytochrome P450 2E1 (CYP2E1) level were measured by 

Western blot. Liver infiltrating macrophages and neutrophils 

were characterized by flow cytometry. Results: Compared with 

WT mice, APAP treated CD36 -/- mice show less liver injury indicated 

by serum ALT level and liver tissue H&E staining. There 

was no significant difference in NAPQI-protein adducts and 

CYP2E1 expression between these two strains. However, less 

IL-1β and CXCL1 mRNA expression were observed in APAP 

treated CD36 -/- mice as well as infiltrating macrophages and 

neutrophils. Conclusion: APAP treated CD36 -/- mice have less 

liver damage than WT mice which correlates with the extent of 

inflammation. CD36 could be a new target to reduce APAP-induced 

liver injury. 

Disclosures: 

The following authors have nothing to disclose: chen zhang, Xianmin Mu, Che 

Xu, Shi Hu, Yuanfang Lu, Qiang You 

1911 

Withaferin-A Reduces Acetaminophen-Induced Liver 

Injury in Mice 

Ravirajsinh Jadeja 1 , Nathalie H. Urrunaga 2 , Suchismita Dash 2 , 

Neeraj K. Saxena 2 , Sandeep Khurana 1 ; 1 Gastroenterology and 

Hepatology, Georgia Regents University, Augusta, MD; 2 Gastroenterology 

and Hepatology, University of Maryland School of Medicine, 

Baltimore, MD 

Acetaminophen (APAP) overdose induces oxidative stress that 

leads to hepatocyte necrosis. Withaferin-A (WA), a lactone, 

has anti-oxidant activities however, its therapeutic potential 

in APAP hepatotoxicity is unknown. The aim of our study was 

to assess the therapeutic potential of WA in a mouse model 

that mimics APAP-induced liver injury (AILI) in humans. Overnight 

fasted C57BL/6NTac male mice (56 wk. old) received 

200 mg/kg APAP intraperitoneally (i.p.). An hour later mice 

were treated with 40 mg/kg WA or vehicle i.p., and euthanized 

at 4 and 16 h; their livers were harvested and serum collected 

for analysis. At 4 h, compared to vehicle-treated mice, 

WA-treated mice had reduced serum ALT levels (P < 0.05), 

hepatocyte necrosis (P


bumin (HNA). Out of the 354 subjects, 68 were analyzed for 

HSA functional changes. We examined the radical scavenging 

activity of purified HSA by monitoring the absorbance of 

diphenylpicrylhydrazyl (DPPH) radicals and the affinity of the 

purified HSA using ketoprofen. Analysis of variance (ANOVA) 

followed by the Tukey’s method, Jonckheere-Terpstra (JT) test, 

receiver operating characteristic curves (ROC), and Pearson’s 

correlation were used for statistical analysis. Results: The radical 

scavenging ability and binding capacity of HSA significantly 

deteriorated with the severity of the disease (P


*Median [Min, Max]; ** P-values from Fisher’s Exact for categorical 

variables, Wilcoxon-Mann-Whitney for continuous variables 

Disclosures: 

Dean W. Roberts - Management Position: Acetaminophen Toxicity Diagnostics, 

LLC 



Laura James - Grant/Research Support: NIDDK; Management Position: Acetaminophen 

Toxicity Diagnostics, LLC 

The following authors have nothing to disclose: Jack A. Hinson, Shasha Bai, R. 

Todd Stravitz, Adrian Reuben, Christopher J. Swearingen, Pippa Simpson 

1915 

Understanding the Relationship between Systemic and 

Hepatic Exposure of Obeticholic Acid for the Treatment 

of Liver Disease in Patients with Cirrhosis 

Jeffrey Edwards 1 , Carl LaCerte 1 , Nathalie H. Gosselin 2 , Thomas 

Peyret 2 , J.F. Marier 2 , Alan F. Hofmann 3 , David Shapiro 1 ; 1 Intercept 

Pharmaceuticals, San Diego, CA; 2 Pharsight, a Certara Company, 

Princeton, NJ; 3 Department of Medicine, University of California 


Obeticholic acid (OCA) is a selective and potent farnesoid 

X receptor (FXR) agonist in development for the treatment of 

primary biliary cirrhosis (PBC) and other liver diseases. OCA 

is structurally similar to chenodeoxycholic acid (CDCA) and 

has similar pharmacokinetics (PK), including conjugation and 

enterohepatic circulation. The amidates of OCA activate FXR 

in the intestine and liver, leading to a reduced hepatic bile 

acid pool. Patients with cirrhosis have higher systemic exposure 

of bile acids (x18) while hepatic exposure of bile acids is 

only ~2-fold higher (Fischer et al. Clinica Chimica Acta 1996 

251:173). In OCA-treated patients, systemic OCA exposure 

was significantly higher in those with moderate (x4) and severe 

(x17) hepatic impairment, similar to increases in systemic exposure 

of endogenous bile acids. FXR activation (measured by 

fibroblast growth factor-19) was similar in patients with hepatic 

impairment versus healthy volunteers, consistent with similar 

liver and intestinal OCA exposure. Thus systemic exposure of 

OCA is not reflective of hepatic concentrations of OCA, the 

primary site of action for safety and efficacy. To understand the 

relationship between systemic and hepatic exposure of OCA 

and its pharmacologically active conjugates in patients with 

and without hepatic (cirrhosis) impairment, a physiologic PK 

model of OCA was developed. The PK model for OCA was 

based on a multi-compartment PK model for CDCA (Molino et 

al. Eur J Clin Invest 1986 16:397) and consisted of systemic, 

hepatobiliary, and intestinal systems. The model accounted 

for the pathophysiology associated with cirrhosis including 

decreased hepatic uptake of OCA, portal-systemic shunting, 

decreased physiologic/functional liver volume, and differences 

in amino acid conjugation. The PK model was used to simulate 

the overall mean concentration-time profile of total OCA in 

plasma and liver. There was good agreement between predicted 

and observed increases in systemic exposure of total 

OCA (the sum of OCA and its conjugates) after a single 10 mg 

dose in subjects with mild (x1.4), moderate (x8), and severe 

(x13) hepatic impairment relative to healthy subjects. The predicted 

increases in liver exposure for subjects with mild, moderate, 

and severe hepatic impairment were only 1.1-, 1.5-, 

and 1.7-fold, respectively, relative to healthy participants and 

are similar to the hepatic exposure of bile acids observed in 

patients with cirrhosis. Model-predicted hepatic exposure of 

OCA, consistent with clinical study results, support the safety 

and efficacy of OCA in PBC patients with cirrhosis at the therapeutic 

doses used in non-cirrhotic patients. 

Disclosures: 

Jeffrey Edwards - Employment: Intercept Pharmaceuticals; Stock Shareholder: 


Carl LaCerte - Employment: Amylin Pharmaceuticals; Stock Shareholder: Amylin 


Nathalie H. Gosselin - Employment: Pharsight 

Thomas Peyret - Employment: Pharsight 

Alan F. Hofmann - Board Membership: Vital Therapies, Vital Therapies; Consulting: 

Albireo Pharma, Shire, Intercept Pharma, Relypsa; Stock Shareholder: 

Intercept Pharma 



The following authors have nothing to disclose: J.F. Marier 

1916 

Transcriptional Profiling Of Primary Human Hepatocytes 

Under Hemodynamics And Transport Differentiates Distinct 

Mechanisms Of Drug-Induced Liver Injury. 

Robert Figler 1 , Brett R. Blackman 1 , Charles Qualls 2 , Svetlana 

Marukian 1 , Sol Collado 1 , Mark Lawson 1 , Aaron J. Mackey 1 , 

David Manka 1 , Brian R. Wamhoff 1 , Ajit Dash 1 ; 1 HemoShear Therapeutics, 

Charlottesville, VA; 2 Qualls Preclinical Solutions LLC, 

Thousand Oaks, CA 

Drug induced liver injury (DILI) is a major cause of liver failure, 

drug withdrawal and non-approval. Predicting DILI is challenging 

since drug concentrations used in in vitro hepatocyte 

cultures are often significantly higher than therapeutic plasma 

concentrations, making mechanistic pathway changes difficult 

to interpret. We have described a system using liver-derived 

blood flow parameters to restore primary human hepatocyte 

biology, allowing for culture at close to physiological insulin/ 

glucose conditions and eliciting drug responses at clinically-relevant 

concentrations. We used this system to assess three DILI 

causing drugs, Acetaminophen (APAP), Trovafloxacin (TVX) 

and Chlorpromazine (CPZ), to differentiate their potential 

for toxicity and transcriptomic signatures, at concentrations 

approximating both therapeutic and toxic levels. For instance, 

the toxic concentration of APAP used (1323 μM) was based on 

serum levels that would initiate treatment for acute toxicity using 

the Rumack-Matthew nomogram, and is significantly lower than 

concentrations typically reported in conventional in vitro culture 

systems. Primary hepatocytes from 5 human donors plated in 

the system were exposed to the drugs for 48 hours. Whole 

genome transcriptomics was performed using RNAseq, along 

with functional assays for CYP activity and function. Distinctive 

responses distinguishing the toxicity potential of the drugs at 

a signaling and phenotypic level underscored the differences 

in underlying mechanisms. Cholestasis-related pathways and 

genes were perturbed by CPZ and TVX but not APAP, consistent 

with lack of cholestatic phenotype in clinical reports of 

APAP toxicity. Fibrotic potential, reflected by increased TGF-β 

signaling, and inflammatory changes evidenced by NFκB activation 

were only seen with TVX. Coagulation and complement 

activation was noted only with APAP while CPZ exhibited activation 

of unfolded protein response. Novel findings including 

suppression of TGF-β with APAP, are currently being explored. 

The correlation of the cellular responses to clinical effects 

reported with the drugs, demonstrates the system’s ability to


predict toxicity mechanisms at clinically relevant concentrations 

and differentiate distinct mechanisms of DILI. 

Disclosures: 

Robert Figler - Employment: HemoShear Therapeutics, LLC 



HemoShear LLC 

Svetlana Marukian - Employment: HemoShear LLC 

Mark Lawson - Employment: Hemoshear 

Aaron J. Mackey - Employment: HemoShear, LLC 





The following authors have nothing to disclose: Charles Qualls, Sol Collado 

1917 

Caffeine Confers Hepatoprotection by Preconditioning 

Hepatocytes with Superior Ability to Withstand DNA 

Damage through Regulation of ATM Signaling Pathways 

Priya Gupta 1 , Yogeshwar Sharma 1 , Sanjeev Gupta 2 , Preeti Viswanathan 

2 ; 1 Medicine and Pathology, Marion Bessin Liver Centre, 

Albert Einstein College of Medicine, Bronx, NY; 2 Division of 

Pediatric Gastroenterology, Hepatology and Nutrition, Children’s 

Hospital at Montefiore Medical Center, Albert Einstein College of 

Medicine, Bronx, NY 

Background: Epidemiological studies indicate that regular 

consumption of caffeine decreases severity of chronic hepatitis, 

cirrhosis, etc. However, the molecular basis for this organ 

protective effect of caffeine is unknown. To develop needed 

paradigms for defining hepatoprotective basis of caffeine we 

hypothesized that ATM signaling pathways, which protect cells 

from DNA damage, will be involved since caffeine inhibits 

ATM kinase activity. Methods: We used HuH-7 human hepatocellular 

carcinoma cells for model development, including 

IC50 cytotoxicity with acetaminophen (APAP) using MTT cell 

viability assays, DNA damage by γH2AX staining and Comet 

formation, and a cell subline expressing hATM promoter-driven 

tdTomato reporter for evaluating global pathway-specific 

changes. Cells were studied with or without caffeine preconditioning 

over 5 d. Flow cytometry was performed to analyze cell 

cycling with PI staining and Side Population (SP) was analyzed 

by Hoechst dyes with or without verapamil pretreatment to 

inhibit dye efflux through MDR activity. Results: APAP toxicity 

in HuH-7 cells led to dose-dependent increases in γH2AX 

expression, Comet formation and greater ATM promoter activity. 

When cells were exposed to APAP plus 10 mM caffeine 

overnight, cytotoxicity was worsened, along with further significant 

increases in γH2AX expression, Comet formation and 

ATM promoter activity, along with depletion of cells in S and 

G2/M, p


for the treatment or prevention of drug-induced fulminant liver 

failure. 

Disclosures: 


Masa Asagiri - Grant/Research Support: Astellas Pharma Inc. 

The following authors have nothing to disclose: Kenji Takemoto, Keiko Iwaisako, 

Masatoshi Takeiri, Naruto Noma, Saori Ohmae, Kan Toriguchi, Kazutaka 

Tanabe, Keigo Machida, Kojiro Taura, Shinji Uemoto 

1919 

Statins prevent liver tumor promoting effect of dioxin-like 

environmental toxins in different biological models 

Alvarez Laura; Facultad de medicina UBA, Argentina, Argentina 

Hepatocellular carcinoma (HCC) is the third leading cause of 

cancer death worldwide. Hormonal imbalance plays a key role 

in the development of neoplasms. There is an open relationship 

between HCC and environmental toxins. Hexachlorobenzene 

(HCB) is an environmental pollutant, associated with a broad 

spectrum of harmful effects on human health as alterations in 

thyroid metabolism, neurotoxicity, developmental and carcinogenic 

effects in human and experimental animals. Statins 

reduce the incidence of various tumors. Their anti-tumor activity 

has been related to their pro-apoptotic and anti-angiogenic 

effect and the prevention of metastasis. However, the exact 

mechanism mediating the in vivo anti-tumor effect of statins has 

not been yet fully clarified. The objective of this study was to 

determine: key molecules involved in HCB promotion of liver 

preneoplastic foci in an initiation-promotion model in rats [diethylnitrosamine 

(DEN) (100 mg / kg bw) and HCB (100 mg / kg 

bw)]. 1- We evaluated in rat liver: a) Proliferating cell nuclear 

antigen levels (PCNA) in focal and non-focal areas, (Western 

blot); b) thyroid hormones (TH) concentration, c) deiodinase 

types I (DI) and III (DIII) mRNA levels; c) 3-Hydroxy-3-methylglutaryl-coenzyme 

A reductase (HMGCoAR) (RIA and RT-PCR), 

d) Thioredoxin 1 levels (TRX1) and d) serum cholesterol. 2-In 

Hep-G2 cells, we analyzed the ability of atorvastatin (AT) and 

simvastatin (SM) to reverse the effect of HCB on key molecules 

involved in the proliferative effect of HCB. Hep-G2 cells were 

treated with HCB (5 uM), in the presence and absence of AT 

(10, 20 and 30 mM) and SM (5, 10, 20 uM). We analyzed 

protein levels of: a) PCNA, b) pERK1/2, c) cyclin D1 (CD1); b) 

TRX1; d) TGF-β1 and HMGCoAR mRNA expression (RT-PCR). 

Results: In vivo: HCB increased (60% p≤0,001) PCNA positive 

cells in focal areas (DEN + HCB) vs. DEN. HMGCoAR 

increased 31% (p≤0,01); T 4 

increased 38% (p≤0.01) and T 3 

decreased 37% (p≤0,01). DIII increased 30% (p≤0.01) and 

DI declined 41% (p≤0.01). TRX1 increased 29% (p≤0.01) in 

(DEN + HCB) (p≤0,01). Cholesterol increased 28% (p ≤ 0.05). 

In vitro, the proliferative effect of HCB decreased with AT or 

SM. HMGCoAR decreased 29% and 38% with AT (20 and 

30 mM), and 20% and 31% with SM (10 to 20 mM) respectively. 

TRX 1 and TGF-β1 protein levels decreased with AT (30 

mM) and SM (20 mM, 34%), p ≤ 0.05. Conclusion: AT and 

SM reduced HCB-induced cell proliferation in Hep-G2 cells. 

HMGCoAR, TGF-β1, TRX1 and HT, may be potential target 

molecules for statins mechanism of action on HCC caused by 

environmental toxins. 

Disclosures: 

The following authors have nothing to disclose: Alvarez Laura 

1920 

Drug-Induced Liver Injury Associated With Vemurafenib 

Treatment 

Marie Lou Gacusan Munson, Liat Schwartz Sagi, Roland Morley, 

Wassim Aldairy, Mason Shih, Edwin Tucker; Genentech, South 


Background: Vemurafenib (VEM) is the first-in-class selective 

oncogenic BRAF V600 kinase inhibitor approved for adults 

with unresectable or metastatic melanoma. In preclinical and 

clinical development programs, VEM has been identified to 

cause liver laboratory abnormalities. The general incidence 

of drug-induced liver injury (DILI) is reported to be 1:10,000 

to 1:100,000 (all severity grades) and is usually identified in 

the postmarketing setting after considerable patient (pt) exposure. 

We describe the incidence, characteristics, risk factors, 

specific clinical signature, and clinical course of hepatotoxicity 

with VEM. Methods: The Genentech company safety database 

was searched for medically confirmed, serious cases of hepatic 

adverse events. The definition of DILI was based on the clinical 

chemistry criteria threshold recommended by the International 

DILI Expert Working Group (EWG) (Aithal et al. Clinical Pharmacol 

Ther. 2011;89:806-815). The WHO Global Introspection 

method was used in the assessment of causality. Clinical 

pattern of liver injury was categorized using the R value; severity 

grading was based on the EWG DILI severity index. Results: 

63 cases of DILI attributed to VEM were identified: 46 from clinical 

trials and 17 from spontaneous reports. Median age was 

58 years (range, 28-83); 17 pts were elderly (≥65 years old). 

27 pts were male and 36 were female. The R value was provided 

for 43 pts; the pattern of injury was cholestatic/mixed in 

28 pts and hepatocellular in 15. Median latency was 43 days 

(range, 1-265) and was similar between cholestatic/mixed 

and hepatocellular patterns. The distribution of clinical pattern 

of injury was similar for elderly and younger pts. DILI severity 

was assessed in 40 pts and was mild, moderate, and severe in 

11, 27, and 2 pts, respectively. No cases had fatal outcomes, 

and no cases required liver transplantation. VEM treatment 

interruption was necessary in 46 of 63 pts; dose modifications 

according to guidance in the product label were necessary in 

25 of those 46. The event did not recur in 20 of the 25 pts. 

In 5 cases, treatment interruption/dose modification required 

permanent discontinuation. Conclusion: The crude reporting 

rate of DILI for VEM is 5.13 per 1000 pt-years (95% CI, 3.8- 

6.4) based on estimated 12,262 pt-years’ exposure. A median 

latency of 43 days with a tendency toward a cholestatic/mixed 

pattern was observed, and 2 cases were assessed as severe. 

Risk factors and populations at risk were not identified. The prescribed 

dose modification in the label was helpful in managing 

cases. To our knowledge, this is the first publication describing 

DILI with a BRAF inhibitor. 

Disclosures: 

Marie Lou Gacusan Munson - Employment: Genentech 

Liat Schwartz Sagi - Employment: Genentech-Roche 

Roland Morley - Employment: Roche Genentech; Stock Shareholder: Bristol Myers 

Squibb 

The following authors have nothing to disclose: Wassim Aldairy, Mason Shih, 

Edwin Tucker


1921 

3D culture strategies for improved MSCs-derived 

hepatocyte-like cells: potential toxicological and clinical 

applications 

Madalena Z. Cipriano 1 , Nora Freyer 2 , Fanny Knoespel 2 , Rita Barcia 

3 , Pedro E. Cruz 3 , Helder Cruz 3 , Nuno G. Oliveira 1 , Jorge M. 

Santos 3 , Katrin Zeilinger 2 , Joana P. Miranda 1 ; 1 CBT, Research 

Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade 

de Lisboa. Av. Prof. Gama Pinto, 1649-003 Lisbon, 

Portugal, Lisboa, Portugal; 2 Bioreactor Group, Berlin Brandenburg 

Center for Regenerative Therapies (BCRT), Charité Universitätsmedizin 

Berlin, 13353 Berlin, Germany, Berlin, Germany; 3 ECBio 

S.A., Rua Henrique Paiva Couceiro, N 27, 2700-451-Amadora, 

Portugal, Amadora, Portugal 

Background: Three-dimensional (3D) cultures have emerged 

as promising alternative models for maintaining hepatocyte 

phenotype in short and long-term in vitro cultures by better 

resembling the in vivo environment of the liver. However, 

human primary hepatocytes have limited availability and 

high inter-individual differences. As such, the differentiation 

of human stem cells into hepatocyte-like cells (HLCs) has been 

suggested. Human neonatal mesenchymal stem cells (hnMSCs), 

derived from the umbilical cord tissue, are particularly interesting 

given their proliferation potential, low immunogenic 

and immuno-modulatory properties along with a demonstrated 

HLCs differentiation in monolayer cultures. However a mature 

hepatic phenotype was not yet achieved. In this study, 3D cell 

cultures, namely a spheroid (SC) and a 3D multi-compartment 

membrane bioreactor (BR) system, were tested as an approach 

for improving HLCs maturation in vitro. Methods: A 27-day 

differentiation protocol consisting of the sequential addition 

of cytokines/growth factors was optimized. Firstly, hnMSCs 

(UCX ® ) were differentiated into hepatoblast-like cells in 2D for 

17 days and afterwards cultured under 3D conditions (SC and 

BR) or maintained in 2D as control for maturation over 10 

days. Results: All systems enabled hnMSC differentiation into 

HLCs as shown by positive immunostaining of hepatic markers, 

such as cytoskeleton proteins (CK-18), the transcription 

factor HNF-4α, albumin, the hepatic transporters OATP-C and 

MRP-2 and CYP1A2 and CYP3A4. Additionally, the three culture 

models displayed relevant glucose metabolism, tested by 

the capacity to store glycogen and to release glucose into the 

medium, and the ability to convert ammonia into urea, to produce 

albumin and to perform phase I metabolism. However, 

urea production (5.8 and 8.3 fold induction for the BR and the 

SC, respectively) and EROD activity (4.8 and 3.8 fold induction 

in the BR and the SC, respectively) were both increased in 

3D relative to 2D. Albumin production was also induced 2.9 

fold in the BR when compared to 2D. Finally, cells differentiated 

within BR presented relevant CYP1A2 and 3A4 activities 

when compared with 2D, whereas HLCs from spheroids and 

2D presented similar CYP3A4, 1A1 and 2C9 activity on day 

27. Conclusions: Overall both the SC and BR models improved 

HLCs maturation suggesting its potential for in vitro and clinical 

applications, namely drug toxicity prediction, regenerative 

medicine or external liver support. Additionally, differential 

HLCs phenotypes were observed for each of the 3D models, 

further supporting that the optimal culture system should be 

selected depending on the scientific purpose. 

Disclosures: 

Rita Barcia - Management Position: ECBio 

Pedro E. Cruz - Board Membership: ECBio, S.A. 

Helder Cruz - Board Membership: ECBio 

Jorge M. Santos - Employment: ECBio 

The following authors have nothing to disclose: Madalena Z. Cipriano, Nora 

Freyer, Fanny Knoespel, Nuno G. Oliveira, Katrin Zeilinger, Joana P. Miranda 

1922 

Death and Liver Transplantations Occurring Within Two 

Years of Drug-Induced Liver Injury 

Paul H. Hayashi 1 , Don C. Rockey 2 , Robert J. Fontana 3 , Hans L. 

Tillmann 4 , Neil Kaplowitz 5 , Huiman X. Barnhart 6 , Jiezhun Gu 6 , 

Averell H. Sherker 7 , Naga P. Chalasani 9 , Victor J. Navarro 8 , 

Jawad Ahmad 10 , Jay H. Hoofnagle 7 ; 1 Division of Gastroenterology 

& Hepatology, University of North Carolina, Chapel Hill, NC; 

2 Medical University of South Carolina, Charleston, SC; 3 University 

of Michigan, Ann Arbor, MI; 4 East Carolina University, Greenville, 

NC; 5 University of Southern California, Los Angeles, CA; 6 Duke 

University, Durham, NC; 7 National Institutes of Health, Bethesda, 

MD; 8 Einstein Medical Center, Philadelphia, PA; 9 Indiana University, 

Indianapolis, IN; 10 Mount Sinai Medical Center, New York, 

NY 

Background: While idiosyncratic drug-induced liver injury (DILI) 

is usually reversible, it can lead to liver transplantation (LT) 

and/or death. But the overall mortality rate and risk factors for 

death or LT are not well defined. Aim: To characterize the role 

and clinical features of DILI in deaths and LT cases. Methods: 

All patients in the Drug Induced Liver Injury Network (DILIN) 

who died or underwent LT within 2 years of follow-up were 

identified. Each case was reviewed independently by 2 hepatologists 

who judged whether DILI had a primary, contributory 

or no role in the death or LT. Cases of DILI having a primary 

role were categorized as acute liver failure (ALF: 6 months) or other. 

Cases of DILI having a contributory role were assigned another 

cause of death (e.g. cancer, sepsis). Discrepancies between 

reviewers were resolved by conference call. R-values (ALT/ULN 

÷ ALP/ULN) at DILI onset were used to classify injury patterns 

as hepatocellular (R>5), mixed (R 2-5) or cholestatic (R


The following authors have nothing to disclose: Paul H. Hayashi, Neil Kaplowitz, 

Huiman X. Barnhart, Jiezhun Gu, Averell H. Sherker, Victor J. Navarro, Jawad 

Ahmad, Jay H. Hoofnagle 

1923 

Severe and prolonged jaundice associated with body 

building supplements (BBS): review of 44 cases from the 

Drug-Induced Liver Injury Network (DILIN) 

Andrew Stolz 1 , Victor J. Navarro 2 , Paul H. Hayashi 3 , Naga P. 

Chalasani 4 , Robert J. Fontana 5 , Herbert L. Bonkovsky 6 , Sunil 

Hwang 6 , Jay H. Hoofnagle 7 , David E. Kleiner 8 , Huiman X. Barnhart 

9 ; 1 Medicine, University of Southern California, Los Angeles, 

CA; 2 Medicine, Einstein Medical Center, Philadelphia, PA; 

3 Medicine, University of North Carolina, Chapel HIll, NC; 4 Medicine, 

Indiana University School of Medicine, Indianapolis, IN; 

5 Medicine, University of Michigan, Ann Arbor, MI; 6 Medicine 

and Research, Carolinas Medical Center, Charlotte, NC; 7 Liver 

Disease Research Branch, Division of Digestive Diseases and 

Nutrition, National Institute of Diabetes and Digestive and Kidney 

Diseases, Bethesda, MD; 8 Laboratory of Pathology, National Cancer 

Institute, Bethesda, MD; 9 Duke Clinical Research Institute, Duke 

University, Durham, NC 

Background & Aim: The use of body building supplements 

(BBS) containing anabolic steroids (AS) continues to rise 

despite efforts to curtail their availability. We sought to characterize 

the clinical features and outcomes of presumed AS-induced 

jaundice in patients taking BBS prospectively enrolled 

in the DILIN. Methods: 44 (5%) out of 847 cases of liver injury 

attributed to drug or herbal dietary supplements enrolled 

between Sept 2004 and March 2013 were attributed solely to 

BBS. The presence of various natural and synthetic androgens 

in consumed products was quantified by LC/MS compared 

with 18 known standards with an average assay sensitivity 

[limit of quantitation] ~ 38 fmoles/mg of product. Liver biopsies 

were interpreted by the study hepatopathologist without 

clinical information. Results: All 44 were men, ages 21 to 59 

(mean = 32) years with 81% self-identified as Caucasian and 

the remainder African-American. 2 subjects had chronic HCV 

infection. The BBS products were used for body building and 

purported to contain various AS with different chemical structures 

and commercial names. All were were taken orally in otherwise 

healthy males. Median latency to onset was 1.8 (range 

0.4 to 14) months, but the exact dates of starting and stopping 

were not always available. All 44 cases were jaundiced 

and symptomatic at presentation and 43 patients had pruritus 

that was frequently severe, prolonged and debilitating. Initial 

median bilirubin was 9.8 mg/dL, ALT 168 U/L, Alk P 111 

U/L and INR 1. Subsequently, median ALT fell steadily, while 

Alk P and bilirubin levels increased 2-3 fold before falling. 

Peak median bilirubin in first 6 months was 25.8 (range 7.3- 

63). 64% were hospitalized and 18% had evidence of hepatic 

dysfunction (e.g. elevated INR). Liver biopsy in 22 patients 

reviewed revealed canalicular cholestasis often with only 

mild portal and lobular inflammation, in contrast to the classic 

“bland cholestasis” often described in association with AS. 

Symptomatic hepatic failure was not observed and no patient 

died or required liver transplantation. Seven of the 44 patients 

(16%) had creatinine elevation > 1.5 mg/dL during their episode 

which trended down as the bilirubin improved. Of 14 

BBS products available for analysis, 71% had identifiable AS 

along with other non-defined steroid containing compounds. 

Conclusion: Jaundice associated with BBS containing AS is a 

distinct clinical syndrome marked by severe and prolonged 

cholestasis in previously healthy men but which did not lead 

to hepatic failure or chronic liver injury in this study. AS were 

often identified in BBS despite efforts to limit their availability. 

Disclosures: 








The following authors have nothing to disclose: Andrew Stolz, Victor J. Navarro, 

Paul H. Hayashi, Sunil Hwang, Jay H. Hoofnagle, David E. Kleiner, Huiman X. 

Barnhart 

1924 

Combined Activities of JNK1 and JNK2 in Hepatocytes 

Protect 

Against Toxin-induced Liver Injury 

Francisco Javier Cubero 1 , Wei Hu 1 , Gang Zhao 1 , Miguel Eugenio 

Zoubek 1 , Jin Peng 1 , Yulia A. Nevzorova 1 , Malika Al Masaoudi 1 , 

Lars Bechmann 4 , Mark V. Boekschoten 3 , Michael Muller 3 , Christian 

Preisinger 2 , Nikolaus Gassler 5 , Ali Canbay 4 , Tom Luedde 1 , 

Roger J. Davis 6 , Christian Liedtke 1 , Christian Trautwein 1 ; 1 Department 

of Medicine III, University Hospital Aachen, RWTH Aachen, 

Aachen, Germany; 2 Proteomics Facility, University Hospital, 

RWTH Aachen, Germany, Aachen, Germany; 3 Nutrition, Metabolism 

& Genomics group Nutrition, Wageningen, The Netherlands, 

Wageningen, Netherlands; 4 Department of Gastroenterology and 

Hepatology, University Hospital Duisburg-Essen, Essen, Germany; 

5 Institute of Pathology, Aachen, Germany; 6 Howard Hughes Medical 

Institute and University of Massachusetts Medical School, 

Worcester, Germany 

Background& Aims: c-Jun N-terminal kinase (JNK)1 and JNK2 

are expressed in hepatocytes and have overlapping and 

distinct functions. JNK proteins are activated, via phosphorylation, 

in response to acetaminophen- or CCl 4 

-induced liver 

damage - the level of activation correlates with the degree 

of injury. SP600125, a JNK inhibitor, has been reported to 

block acetaminophen-induced liver injury. We investigated 

the role of JNK in drug-induced liver injury (DILI) in liver tissues 

from patients and in mice with genetic deletion of JNK in 

hepatocytes. Methods: We studied liver sections from patients 

with DILI (due to phenprocoumon, non-steroidal anti-inflammatory 

drugs, or acetaminophen) or autoimmune hepatitis, or 

patients without acute liver failure (controls), collected from a 

DILI Biobank in Germany. Levels of total and activated (phosphorylated) 

JNK were measured by immunohistochemistry and 

immunoblot assays. Mice with hepatocyte-specific deletion of 

Jnk1 (Jnk1 Δhepa ) or combination of Jnk1 and Jnk2 (Jnk Δhepa ), 

as well as Jnk1-floxed C57BL/6 (control) mice, were given 

repetitive CCl 4 

injections to induce fibrosis or acetaminophen 

to trigger toxic liver injury. We performed gene expression 

microarray, and phosphoproteomic analyses to determine 

mechanisms of JNK activity in hepatocytes. Results: Liver samples 

from patients with DILI contained more activated JNK, predominantly 

in hepatocytes, than healthy tissue. Administration 

of acetaminophen to Jnk Δhepa mice produced a greater level 

of liver injury than that observed in Jnk1 Δhepa or control mice, 

based on levels of serum markers and microscopic and histologic 

analysis of liver tissues. Administration of CCl 4 

induced 

significantly stronger hepatic injury in Jnk Δhep mice, based on 

increased inflammation, cell proliferation, and fibrosis progression, 

compared to Jnk1 Δhepa or control mice. Hepatocytes from 

Jnk Δhepa mice given acetaminophen had an increased oxidative 

stress response, leading to decreased activation of AMPK and 

JunD and subsequent necrosis. Administration of SP600125 

before or with acetaminophen protected Jnk Δhepa and control 

mice from liver injury. Conclusions: In hepatocytes, JNK1 and


JNK2 have combined effects in protecting mice from CCl 4 

- and 

acetaminophen-induced liver injury. It is important to study the 

functions of both proteins, rather than just JNK1, to define their 

role during toxic liver injury. JNK inhibition with SP600125 

has off-target effects. 

Disclosures: 



The following authors have nothing to disclose: Francisco Javier Cubero, Wei 

Hu, Gang Zhao, Miguel Eugenio Zoubek, Jin Peng, Yulia A. Nevzorova, Malika 

Al Masaoudi, Lars Bechmann, Mark V. Boekschoten, Michael Muller, Christian 

Preisinger, Nikolaus Gassler, Ali Canbay, Tom Luedde, Roger J. Davis, Christian 

Liedtke 

1925 

Translocation of Iron from Lysosomes to Mitochondria 

during Acetaminophen-Induced Toxicity in Mouse 

Hepatocytes 

Jiangting Hu 1 , Andaleb Kholmukhamedov 1 , Hartmut Jaeschke 2 , 

John J. Lemasters 1 ; 1 Medical University of South Carolina, Charleston, 

SC; 2 University of Kansas Medical Center, Kansas city, KS 

Background: Acetaminophen (APAP) overdose causes hepatotoxicity 

involving mitochondrial dysfunction and the mitochondrial 

permeability transition (MPT). Reactive oxygen species 

(ROS) play an important role in APAP-induced hepatotoxicity. 

Iron is a critical catalyst for ROS formation. Previous studies 

show that disrupted lysosomes release ferrous iron (Fe 2+ ) 

into the cytosol during APAP hepatotoxicity, which triggers 

the MPT and cell killing. Here, our Aim was to investigate 

the role of lysosomal iron mobilization into mitochondria 

during APAP-induced toxicity to mouse hepatocytes. Methods: 

Hepatocytes were isolated from fasted male C57BL/6 mice. 

Necrotic cell killing was assessed by propidium iodide fluorimetry. 

Mitochondrial membrane potential was visualized by 

confocal microscopy of rhodamine 123 (Rh123) or tetramethylrhodamine 

methylester (TMRM). Chelatable Fe 2+ was monitored 

by quenching of calcein (cytosol) and mitoferrofluor (MFF, 

mitochondria). Results: Starch-desferal (1 mM, a lysosomally 

targeted iron chelator) and Ru360 (100 nM, an inhibitor of the 

mitochondrial calcium uniporter [MCU]) decreased cell killing 

after APAP (10 mM) by 50% and 25% at 10 h, respectively. 

Progressive quenching of calcein and MFF began after ~4 h, 

signifying increased cytosolic and mitochondrial chelatable 

Fe 2+ . Mitochondria then depolarized after ~10 h. Dipyridyl, a 

membrane-permeable iron chelator, dequenched calcein and 

MFF fluorescence. Starch-desferal, but not Ru360, suppressed 

cytosolic calcein quenching, whereas both starch-desferal and 

Ru360 suppressed mitochondrial MFF quenching and mitochondrial 

depolarization. Conclusion: Release of chelatable 

Fe 2+ from lysosomes followed by uptake into mitochondria via 

MCU occurs during APAP hepatotoxicity, which triggers the 

MPT and cell killing. Ru360 prevents the increase of mitochondrial 

but not cytosolic Fe 2+ after APAP but is not as protective as 

starch-desferal, which prevents Fe 2+ increases in both compartments. 

Thus, increased Fe 2+ in the cytosol may also contribute 

to APAP hepatotoxicity. 

Disclosures: 

John J. Lemasters - Consulting: Novo Nordisk 

The following authors have nothing to disclose: Jiangting Hu, Andaleb 

Kholmukhamedov, Hartmut Jaeschke 

1926 

Azathioprine And 6-Mercaptopurine Induced Liver 

Injury 

Bjornsson S. Einar 1,2 , Jiezhun Gu 3 , David E. Kleiner 2 , Naga P. 

Chalasani 4 , Paul H. Hayashi 5 , Jay H. Hoofnagle 6 ; 1 The NAtional 

University Hospital of Iceland, Reykjavik, Iceland; 2 NIH, Bethesda, 

MD; 3 3Duke Clinical Research Institute, Durham, NC; 4 Indiana 

University School of Medicine, Indinapolis, IN; 5 University of 

North Carolina, Chapel Hill, NC; 6 NIDDK, NIH, Bethesda, MD 

Background: Most information of regarding the drug induced 

liver injury (DILI) from azathioprine (Aza) and 6-mercaptopurine 

(6-MP) comes from isolated case reports and the clinical 

features and frequency of outcomes is not well defined. The 

aims of this study were to better define the clinical, biochemical 

and histologic features of Aza and 6-MP induced liver 

injury. Methods: Patients with thiopurine hepatotoxicity from 

the DILIN Prospective Study were identified. Patients enrolled 

between 2004 and 2014 underwent structured assessment of 

potential competing etiologies and analysis of clinical features. 

Results: 22 patients (12 attributed to Aza, 10 to 6-MP, none 

to thioguanine) were identified who had probable (n=8), very 

likely (n=12) or definite (n=2) causality scores. The median 

age was 55 years, and 68% were female. Inflammatory bowel 

disease was the indication in 55%, and median dose was 150 

mg daily (range 25-300 mg). The duration of therapy before 

onset varied widely (median 75, range 3 to 2584 days). However, 

injury often arose after a dose escalation (59%) and the 

median time of onset after a last dose increase was 44 days 

(range 3 to 254 days). Overall 73% of cases were icteric. The 

median initial ALT was 210 U/L, Alk P 151 U/L and bilirubin 

7.4 mg/dL. Median peak bilirubin levels were 13.4 mg/ 

dL and INR 1.3. Icteric and anicteric cases differed in their 

typical clinical features. Cases without jaundiced had minimal 

symptoms and a hepatocellular pattern of enzyme elevations, 

whereas jaundiced cases had a self-limited cholestatic 

hepatitis with minimal or modest elevations in enzyme levels. 

Available biopsies (7 cases) showed cholestatic hepatitis; 3 

also showed evidence of nodular regeneration. There was little 

fibrosis except in the patient with pre-existing cirrhosis. There 

were no major differences between Aza and 6-MP cases. One 

The patient with pre-existing cirrhosis underwent emergency 

liver transplantation, all others resolved the injury clinically, 

although one patient had moderate Alk P elevations 2 years 

after onset. Conclusions: Nearly three-quarters of thiopurine-induced 

liver injury presents with self-limited cholestatic hepatitis. 

Injury often arises within a few months of dose increase, 

highlighting the importance of monitoring liver tests after dose 

escalaction. The prognosis of Aza and 6-MP related liver injury 

is favorable except in patients with pre-existing cirrhosis. 

Disclosures: 



The following authors have nothing to disclose: Bjornsson S. Einar, Jiezhun Gu, 

David E. Kleiner, Paul H. Hayashi, Jay H. Hoofnagle 

1927 

Gender-specific glucuronidation of acetaminophen in 

htgUGT1A WT mice 

Anja Winkler, Sandra Kalthoff, Christian P. Strassburg; Clinics and 

Policlinic I, University Hospital Bonn, Bonn, Germany 

Aims: Acetaminophen (APAP) is a commonly used over-thecounter 

drug with analgesic as well as antipyretic properties 

and at low doses is primarily metabolized through sulfation 

and glucuronidation, while only a small fraction is oxidized to 

the reactive metabolite N-acetyl-p-benzoquinone (NAPQI) by


cytochrome P450 enzymes. At supratherapeutic doses saturation 

of the sulfation and glucuronidation pathways lead to an 

increase of NAPQI and therefore to APAP-mediated hepatotoxicity 

potentially causing acute liver failure (ALF). Previous studies 

have already reported gender-related differences in APAP 

metabolism with females displaying relative resistance towards 

APAP-induced hepatotoxicity. However, the exact molecular 

mechanisms involved are not fully understood. Aim of this study 

was therefore to evaluate possible gender-related differences 

in glucuronidation of APAP in a humanized tgUGT1A mouse 

model. Methods: Male and female htgUGT1A WT mice were 

intraperitoneally (i.p.) treated with acetaminophen (APAP, 5 

mg/kg) or vehicle for three consecutive days. Expression of 

UGT1A genes and transcription factors (TF) HNF-4α, PXR and 

ESR were analyzed by Taqman-PCR. Results: Assessment of 

hepatic UGT1A mRNA expression in APAP-treated animals 

revealed a 10-, 36- and 11-fold increase of UGT1A1, UGT1A6 

and UGT1A9 in female mice contrasting an only 7-, 9- and 

10-fold induction of the aforementioned isoforms in males. 

Additionally, female mice exhibited significant increases of 

UGT1A1 (3.2-, 7.4-fold) and UGT1A6 (5.6-, 4.8-fold) in both 

jejunum and colon, whereas in males upregulation of intestinal 

UGT1A genes was only observed in colon (UGT1A1: 6.3-fold; 

UGT1A6: 7.2-fold). Interestingly, investigation of TFs in APAPtreated 

htgUGT1A WT mice revealed upregulation of ESR and 

PXR mRNA in livers of females, which was reduced in male 

mice. Moreover, HNF4-a expression was downregulated in 

males. Conclusions: Analyses of hepatic as well as intestinal 

UGT1A genes relevant for APAP glucuronidation uncovers significant 

differences in UGT1A mRNA expression between male 

and female mice following treatment with acetaminophen. Particularly 

APAP-mediated upregulation of hepatic UGT1A6 was 

profoundly reduced in male htgUGT1A WT mice in comparison 

to their female counterparts. These data suggest a possible 

role of glucuronidation for gender-related distinctions in 

susceptibility towards APAP-mediated hepatotoxicity and clinically 

observed sex-related differences in acetaminophen pharmacokinetics 

and –dynamics. Furthermore, reduced hepatic 

expression of the TFs ESR, PXR and HNF4-a represents a possible 

molecular mechanism for differential UGT1A regulation 

in males 

Disclosures: 



The following authors have nothing to disclose: Anja Winkler, Sandra Kalthoff 

1928 

Drug Induced Liver Injury (DILI) in the East 

Sarah E. Low 1 , Kieron B. Lim 1 , Seng Gee Lim 1,2 ; 1 Gastroenterology 

& Hepatology, National University Health System, Singapore, 

Singapore; 2 Yong Loo Lin School of Medicine, National University 

of Singapore, Singapore, Singapore 

Background Is Drug Induced Liver Injury (DILI) different in the 

East compared with the West? There are only a handful of 

population studies worldwide estimating crude incidence rates 

of 3-19 of 100,000 inhabitants. Asians have different pharmacogenetic 

profiles with different disease patterns, susceptibility 

and hence handle drugs differently. There is widespread use 

of herbal and alternative medicines (HM), the safety of which 

is poorly defined. Aims To review existing literature on prevalence 

of DILI in the East compared to the West. We also 

seek to examine differences in frequency of causative agents 

of DILI in the East and postulate why. Methods A Medline 

search with “Drug-Induced Liver Injury[Mesh] OR Hepatotoxicity 

AND” was undertaken. Countries included:Singapore, 

Malaysia, Indonesia, Thailand, India, Sri Lanka, 

Vietnam, China, Korea, Taiwan, Pakistan, Cambodia, Japan, 

Hong Kong, Nepal, Mongolia, Laos, Philippines. Results Anti 

tuberculosis (TB) drugs (57% of DILI in India) and HM were most 

commonly implicated. Of 290 articles, 43.5% were on TB DILI. 

Although TB is more prevalent in Asia, Asians appear more 

susceptibile to TB DILI, with twice as much prevalence, 6.1% 

versus 3.1%, likely due to polymorphisms of drug metabolizing 

genes (NAT2, CYP2D1, GSTM1, GSTT1). Despite under 

reporting, proportion of DILI from HM ranged from 18%-40% 

with incidence of DILI in HM consumers to be 0.7%-1.5%. Hepatotoxicity 

from Amoxicillin Clavulanate was rarely reported in 

the East. Severity of intrinsic hepatotoxicity of Acetaminophen 

is attenuated in the East, with virtually no cases of liver failure 

or death reported. Conclusion The profile of DILI appears to 

be different in the Asia compared to the West, with primarily 

TB DILI and Herbal DILI, and rarely Amoxicillin Clavulanate or 

Acetaminophen in contrast to the West 

Disclosures: 

Kieron B. Lim - Advisory Committees or Review Panels: Gilead, Abbvie, Sirtex; 

Consulting: Novartis; Grant/Research Support: Astellas, Bayer 






The following authors have nothing to disclose: Sarah E. Low 

1929 

Application of the Drug-Induced Liver Injury Expert 

Working Group Criteria in the Assessment of Hepatotoxicity 

With the BRAF Inhibitor Vemurafenib 

Marie Lou Gacusan Munson, Liat Schwartz Sagi, Mason Shih, 

Edwin Tucker; Genentech, South San Francisco, CA 

Background: Vemurafenib (VEM) is the first BRAF inhibitor 

approved by the US Food and Drug Administration (FDA) and 

the European Medicines Agency (EMA) for patients with metastatic 

melanoma. At the time of its approval, based on a safety 

database of ~500 patients (EMA. Public Assessment Report 

on Vemurafenib. 2011), VEM was known to cause liver laboratory 

abnormalities. After 2 y of commercial use, Hy’s law 

cases were identified, prompting a comprehensive hepatotoxicity 

risk review of VEM. The international Drug-Induced Liver 

Injury (DILI) Expert Working Group (EWG) DILI criteria (Aithal 

et al. Clinical Pharmacol Ther. 2011;89:806-815), with an 

EBM 2B level of evidence, were used to evaluate VEM. The


aims were to (1) assess the diagnostic yield of the EWG clinical 

chemistry criteria for DILI when applied to individual case 

safety reports of VEM, (2) describe the limitations of the EWG 

criteria in real-world application and (3) present the modifications 

implemented. Methods: We conducted a retrospective 

review and analysis of the EWG algorithm and diagnostic criteria 

for identifying and evaluating DILI cases for VEM. Results: 

228 serious, medically confirmed cases with hepatic adverse 

events were included; 81 met the DILI EWG criteria threshold: 

a diagnostic yield of 35.5%. The EWG algorithm was modified 

at 3 levels: (1) The CTCAE severity grading was substituted 

for actual laboratory values. (2) Cases that did not meet the 

DILI criteria but were reported as hepatic failure or had fatal 

outcomes were included during causality assessment so clinically 

significant cases were not missed. (3) To allow flexibility 

with missing data, the WHO global introspection method for 

causality assessment was used instead of the CIOMS Roussel 

Uclaf Causality Assessment Method (RUCAM). Conclusions: 

The DILI EWG clinical chemistry criteria and algorithm provide 

an objective and systematic approach to the assessment of DILI 

within the postmarketing setting. The modifications proposed 

herein address the inadequate case information from spontaneous 

sources. 

Disclosures: 

Marie Lou Gacusan Munson - Employment: Genentech 

Liat Schwartz Sagi - Employment: Genentech-Roche 

The following authors have nothing to disclose: Mason Shih, Edwin Tucker 

1930 

Minocycline hepatotoxicity: Clinical characterization and 

identification of HLA- B*35:02 as a risk factor 

Thomas J. Urban 1,2 , Paola Nicoletti 3 , Naga P. Chalasani 4 , Jose 

Serrano 5 , Andrew Stolz 6 , Ann K. Daly 7 , Guruprasad P. Aithal 8 , 

John F. Dillon 9 , Huiman X. Barnhart 10 , Paul B. Watkins 2 , Robert 

J. Fontana 11 ; 1 Eshelman School of Pharmacy, University of North 

Carolina at Chapel Hill, Chapel Hill, NC; 2 Hamner Institute for 

Drug Safety Sciences, Durham, NC; 3 Center for Computational 

Biology and Bioinformatics, Columbia University, New York, 

NY; 4 Indiana University School of Medicine, Indianapolis, IN; 

5 National Institute of Diabetes Digestive and Kidney Diseases, 

Bethesda, MD; 6 University of Southern California, Los Angeles, 

CA; 7 University of Newcastle, Newcastle, United Kingdom; 8 Nottingham 

University, Nottingham, United Kingdom; 9 University of 

Dundee, Dundee, United Kingdom; 10 Duke University, Durham, 

NC; 11 University of Michigan, Ann Arbor, MI 

Prolonged minocycline use has been associated with rare 

instances of hepatotoxicity that can present with prominent 

autoimmune features in previously healthy individuals. Prior 

genome-wide association studies (GWAS) have demonstrated 

human leukocyte antigen (HLA) polymorphisms that are associated 

with DILI susceptibility to various agents. The aim of this 

study was to identify potential genetic determinants of minocycline 

DILI in a well-phenotyped cohort of patients. METHODS: A 

total of 25 Caucasian patients (22 from DILIN, 3 from iDILIC) 

with DILI due to minocycline underwent genome-wide genotyping 

and were compared to population controls. Coding 

sequences of HLA alleles were imputed based on SNP genotypes 

in the HLA region. Verification of HLA carrier status was 

undertaken using sequence-based HLA typing and included an 

additional 2 DILIN cases that were not included in the GWAS. 

RESULTS: Amongst the 26 cases from DILIN with complete clinical 

information, 73.1% were female with a median age of 

19.5 years (range: 15 to 61). The median latency from drug 

start to DILI onset was 330 days (range: 15 to 1350) with 70% 

of cases presenting after 6 months of minocycline exposure. At 

DILI onset, 50% were jaundiced, 19% reported rash, and 31% 

had fever but only 4% had eosinophilia. At presentation, 82% 

had acute hepatocellular liver injury, median ALT of 1212 IU/L 

(range: 70 to 2500), median bilirubin of 4.5 mg/dl (range: 

0.2 to 16.7), 92% had a + ANA and 29% had a + SmAb. 

During follow-up, 46% were treated with corticosteroids and 

the median time to ALT normalization was 93 days. None of 

the subjects died or required a liver transplant but 25% had evidence 

of chronic liver injury at 6 months of follow-up. GWAS 

revealed a strong association between HLA-B*35:02 and risk 

for minocycline-DILI, with a carrier frequency of 16% in DILI 

cases compared to 0.6% in population controls (Odds Ratio: 

29.6, 95% CI: 9.0-97.5, p=2.5 x 10 -8 ). Verification of HLA- 

B*35:02 imputation was confirmed by sequence-based HLA 

typing. HLA-B*35:02 carriers had a lower R-value at presentation 

(9 vs. 26, p=0.018) but otherwise had similar presenting 

features and outcomes compared to non-carriers. HLA-B*35:02 

has been shown to hasten progression to AIDS among HIV 

patients, but has not previously been associated with any drugor 

liver-related phenotypes. CONCLUSIONS: Long-term treatment 

with minocycline can result in infrequent severe acute 

hepatocellular liver injury with prominent autoimmune features 

in young female patients. HLA-B*35:02 is a rare HLA allele 

that is strongly associated with minocycline-DILI, and may be a 

useful diagnostic marker of this serious adverse event. 

Disclosures: 








The following authors have nothing to disclose: Thomas J. Urban, Paola Nicoletti, 

Jose Serrano, Andrew Stolz, Ann K. Daly, Guruprasad P. Aithal, John F. Dillon, 

Huiman X. Barnhart 

1931 

Liver Disorders that can Masquerade as Drug-Induced 

Liver Injury 

Don C. Rockey 1 , Paul H. Hayashi 2 , Hans L. Tillmann 3 , Jiezhun 

Gu 3 , Marwan Ghabril 4 , Jawad Ahmad 5 , Andrew Stolz 7 , Robert J. 

Fontana 8 , Jay H. Hoofnagle 6 ; 1 Medical University of South Carolina, 

Charleston, SC; 2 University of North Carolina, Chapel Hill, 

NC; 3 Duke University, Durham, NC; 4 Indiana University, Indianapolis, 

IN; 5 Mount Sinai, New York, NY; 6 NIH, Bethesda, MD; 

7 University of Southern California, Los Angeles, CA; 8 University of 

Michigan, Ann Arbor, MI 

Background: Drug-induced liver injury (DILI) is a diagnosis of 

exclusion, and requires a high degree of clinical suspicion. 

The aim of this study was to better understand the features


and diagnoses of patients who were initially thought to have 

DILI, which was later judged to be unlikely. Methods: Between 

2004 and 2014, the Drug Induced Liver Injury Network (DILIN) 

Prospective Study recruited and assessed 1405 patients suspected 

to have DILI. All patients underwent evaluation for 

viral hepatitis (A, B, C, CMV, E, HSV), autoimmune hepatitis, 

pancreaticobiliary disease, and alcohol. Cases under went a 

structured causality assessment at 6 months of follow-up to be 

assigned into 5 categories of likelihood (definite, >95%; highly 

likely, 75-94%; probable, 50-74%, possible, 25-49%, unlikely, 


1933 

Genome-wide investigation identifies common SNPs on 

Chr9p22.2 as risk factors for liver injury due to sulfamethoxazole/trimethoprim 

Thomas J. Urban 1,2 , Paola Nicoletti 3 , Robert J. Fontana 4 , Andrew 

Stolz 5 , Ann K. Daly 6 , Guruprasad P. Aithal 7 , M. I. Lucena 8 , Huiman 

X. Barnhart 9 , Paul Watkins 2 , Naga P. Chalasani 10 ; 1 Eshelman 

School of Pharmacy, University of North Carolina at Chapel 

Hill, Chapel Hill, NC; 2 Hamner Institute for Drug Safety Sciences, 

Durham, NC; 3 Columbia University, New York, NY; 4 University 

of Michigan, Ann Arbor, MI; 5 University of Southern California, 

Los Angeles, CA; 6 University of Newcastle, Newcastle upon Tyne, 

United Kingdom; 7 Nottingham University, Nottingham, United 

Kingdom; 8 University of Malaga, Malaga, Spain; 9 Duke University, 

Durham, NC; 10 Indiana University School of Medicine, Indianapolis, 

IN 

BACKGROUND &AIM: Treatment with the antibiotic combination 

sulfamethoxazole/trimethoprim (SMZ/TMP) can result in a 

rare but serious liver toxicity which typically presents with immunoallergic 

features. Currently there are no genetic or non-genetic 

predictors of toxicity. We aimed to uncover genetic risk 

factors associated with SMZ/TMP DILI. METHODS: Genomewide 

genotyping was performed on 27 individuals of European 

ancestry who experienced liver injury from SMZ/TMP, including 

22 DILIN patients and 5 from iDILIC/iSAEC. Genotype 

frequencies were compared to >5,000 population controls. 

Targeted genotyping by TaqMan assay was performed for validation 

in 33 cases (19 cases included in the GWAS and 14 

new cases not included in the GWAS). RESULTS: GWAS was 

conducted on 27 individuals with SMZ/TMP DILI with mean 

age 47 years, 48% females, and the pattern of liver injury was 

hepatocellular in 41% and cholestatic in 30%. Their peak ALT 

(mean ± sd) were 714 ± 542 U/L and peak bilirubin 11.5± 

10.1 mg/dL. The GWAS revealed a set of common SNPs in an 

intergenic region on Chr9p22.2 that showed strong evidence 

of association with SMZ/TMP-DILI. The minor allele frequency 

(MAF) of the risk allele was 26% in SMZ/TMP-DILI cases, vs. 

4.6% in controls (OR: 9.0, 95%CI: 4.3-19.1, p = 5 X 10 -9 ). 

Validation cohort consisted of 33 European Caucasians (19 

from the GWAS plus 14 independent replication samples) with 

mean age 46 years, 55% females and the pattern of liver injury 

was hepatocellular in 28% and cholestatic in 32%. Their peak 

ALT values were 745 ± 695 U/L and bilirubin 13.1± 10.1 mg/ 

dL. Targeted assays confirmed the genotypes as measured by 

the GWAS arrays, and further identified 4 additional carriers 

among 14 new cases in the validation cohort (MAF=14.3%, 

p < 0.05). However, the phenotypic characteristics of DILI in 

patients who carried the risk alleles (age, sex, latency to onset, 

immunoallergic features, peak laboratory test values) were not 

significantly different from those who lacked the risk alleles at 

Chr9p22.2. CONCLUSIONS: Common SNPs on Chr9p22.2 

are strongly associated with risk for SMX/TMP-DILI. These 

SNPs are not located near any known protein-coding gene or 

miRNA, and have not previously been associated with mRNA 

expression of any gene. Further studies will be necessary to 

understand the mechanism underlying the association, and to 

extend these studies to other hypersensitivity reactions to SMZ/ 

TMP as well as to DILI caused by other sulfonamides. 

Disclosures: 





The following authors have nothing to disclose: Thomas J. Urban, Paola Nicoletti, 

Andrew Stolz, Ann K. Daly, Guruprasad P. Aithal, M. I. Lucena, Huiman X. 

Barnhart, Paul Watkins 

1934 

Co-activation of AKT and c-Met triggers rapid hepatocellular 

carcinoma development via mTORC1/FASN 

pathway in mice 

Junjie Hu 2,1 , Lei Li 3,1 , Li Che 1 , Diego Calvisi 4 , Xin Chen 1 ; 1 Bioengineering 

and Therapeutic Sciences, UCSF, San Francisoco, 

CA; 2 School of Pharmacy, Hubei University of Chinese Medicine, 

Wuhan, China; 3 School of Pharmacy, Huazhong university of science 

and technology, Wuhan, China; 4 Institude of Pathology, university 

of greifswald, Greifswald, Germany 

Background: Activation of the AKT/mTOR cascade and overexpression 

of c-Met have been implicated in the development 

of human hepatocellular carcinoma (HCC). Methods: To elucidate 

the functional crosstalk between the AKT and c-Met pathways, 

we generated a model characterized by the combined 

expression of activated AKT and c-Met in the mouse liver using 

hydrodynamic transfection. Results: We found that co-expression 

of AKT and c-Met results in faster liver tumor development 

when compared with mice overexpressing AKT alone, whereas 

c-Met alone does not lead to tumor formation. At the molecular 

level, liver tumors induced by AKT/c-Met display activation of 

AKT/mTOR and Ras/MAPK cascades as well as increased 

lipogenesis and glycolysis. Mechanistically, AKT/c-Met driven 

hepatocarcinogenesis was found to be mTORC1-dependent, as 

Raptor genetic deletion abrogates tumor development in AKT/ 

c-Met mice. Furthermore, since increased de novo synthesis of 

fatty acid is a pivotal metabolic event downstream of mTORC1, 

we determined the requirement of lipogenesis in AKT/c-Met 

driven liver tumor development using conditional Fatty Acid 

Synthase (FASN) knockout mice. Of note, hepatocarcinogenesis 

induced by AKT/c-Met was fully inhibited by FASN ablation. 

In human HCC samples, coordinated expression of FASN, 

c-Met, activated AKT and ERK proteins was detected in a subgroup 

of biologically aggressive tumors. Conclutions: Our study 

demonstrates that co-activation of AKT and c-Met induces HCC 

development that depends on the mTORC1/FASN pathway. 

Suppression of mTORC1 and/or FASN might be highly detrimental 

for the growth of human HCC subsets characterized by 

concomitant induction of the AKT and c-Met cascades. 

Disclosures: 

The following authors have nothing to disclose: Junjie Hu, Lei Li, Li Che, Diego 

Calvisi, Xin Chen 

1935 

Classification of tumors based on the integrated profile 

of genetic and epigenetic alterations and the biological 

behavior of human hepatocellular carcinoma 

Naoshi Nishida 1 , Toshimi Kaido 2 , Masatoshi Kudo 1 ; 1 Department 

of Gastroenterology and Hepatology, Kinki University School of 

Medicine, Osaka-sayama, Japan; 2 Department of Surgery, Graduate 

School of Medicine Kyoto University, Kyoto, Japan 

Aims: We tried to classify HCCs based on the integrated 

genetic and epigenetic profile, and examined the association 

between the classification and biological characteristics 

such as high metastatic potential. Methods: (1) A total of 121 

HCCs were examined for p53, β-catenin and TERT promoter 

mutation, methylation of 8 tumor suppressor genes (TSGs) 

and hypomethylation on 3 repetitive DNA sequences (RSs), 

and fractional allelic loss (FAL) as a surrogate marker of chromosomal 

instability (CI) using direct sequencing, MethyLight, 

and microsatellite analyses with 200 markers, respectively. A 

degree of TSG methylation and hypomethylation of RSs was 

categorized as “extensive” vs. “limited” and “significant” vs. 

“slight” by hierarchical clustering, respectively. Each tumor was


classified according to the integrated molecular profiles using 

corresponding and hierarchical clustering analyses, and their 

relationship to the clinicopathological backgrounds were examined. 

(2) Molecular profiles of 44 HCCs from the patients who 

underwent liver transplantation (LT) were determined based 

on the mutation, methylation and microsatellite analyses. We 

tried to examine the specific molecular features that related to 

the extrahepatic recurrence after LT of HCC. Results: (1) We 

successfully classified HCCs in to 4 subgroups based on the 

genetic and epigenetic alterations. The subgroup A1 (S-A1) 

was characterized by high frequencies of extensive TGS methylation, 

β-catenin and TERT promoter mutation; however, the 

molecular characteristics of the subgroup B2 (S-B2) were the 

presence of significant hypomethylation on RSs and p53 mutation 

accompanied by CI, which was mutually exclusive to that 

of S-A1. Similarly, the subgroup A2 (S-A2) was characterized 

by the presence of all the alterations examined. On the other 

hand, genetic and epigenetic alterations were rare in HCCs in 

the subgroup B1 (S-B1). Characteristics related to tumor progression, 

such as high serum AFP (> 200 ng/ml), larger tumor 

size (> 3 cm), presence of vascular invasion, and dedifferentiated 

tumor were more frequently observed in HCCs belonging 

to S-A2 and S-B2 than those in S-A1 and S-B1. (2) Among 44 

HCC patients after LT, 6 showed metastatic recurrence of HCC 

after LT and 38 had not shown any recurrence for more than 2 

years. Five of 6 HCC patients (83%) showing recurrence carried 

tumors with S-B2 molecular characteristics, whereas only 

11 of 38 HCCs in non-recurrence group (29%) showed S-B2 

characteristics. Conclusion: Integrated genetic and epigenetic 

profiles are associated with tumor characteristics and predict 

mild or aggressive behavior of HCCs. 

Disclosures: 

Masatoshi Kudo - Advisory Committees or Review Panels: Bayer HealthCare; 

Grant/Research Support: Bayer HealthCare 

The following authors have nothing to disclose: Naoshi Nishida, Toshimi Kaido 

1936 

Baicalein targets liver tumor initiating stem cell-like cells 

resistant to mTORC1 inhibition 

Raymond Wu 1 , Ramachandran Murali 2 , Yasuaki Kabe 3 , Keigo 

Machida 1 , Clay Wang 4 , Stan G. Louie 4 , Hidekazu Tsukamoto 1 ; 

1 Southern California Research Center for ALPD and Cirrhosis, Keck 

School of Medicine, University of Southern California, Los Angeles, 

CA; 2 Cedars-Sinai Medical Center, Los Angeles, CA; 3 Keio University 

School of Medicine, Tokyo, Japan; 4 Department of Pharmacology 

and Pharmaceutical Sciences, School of Pharmacy of the 

University of Southern California, Los Angeles, CA 

[Introduction] mTORC1 inhibitors are tested for the treatment 

of HCC based on hyperactive mTOR in this malignancy. However, 

evidence indicates mTORC1 inhibitors may promote 

CD133 upregulation and chemoresistance. CD133+ tumor initiating 

stem cell-like cells (TIC) isolated from mouse liver tumors 

(PNAS 106:1548; JCI 123:2832) are chemoresistant, and 

identification of an approach to abrogate this resistance is critical. 

[Objective] We searched for a natural compound which 

rescinds TIC’s resistance to mTORC1 inhibition and improves 

chemotherapy outcome. [Methods and Results] HPLC/MS and 

NMR analyses combined with TIC lethality assay, identified 

baicalein (BC) as a bioactive compound in the herbal medicine 

Yan Gang Wan shown to suppress DEN-induced liver 

tumorigenesis in mice. BC dose-dependently inhibits TIC self-renewal 

shown by spheroid formation and clonogenic assay 

and induces TIC apoptosis at IC50=9.5mM while having no 

adverse effects on mouse primary hepatocytes. BC represses 

stemness genes (Nanog, Sox2) while upregulating hepatocyte 

differentiation genes (Alb, Cyp7a1). Structure-function analysis 

reveals –OH groups on C5 and C7 are required for TIC killing. 

TIC which are resistant to rapamycin (2mM) and temsirolimus 

(~4.5mM), are killed ~90% when co-treated with BC (10mM). 

BC also enhances TIC cytotoxicity by sorafenib (3.3mM) and 

doxorubicin (1mM), causing ~95% elimination. BC suppresses 

autophagosome formation determined by LC3II IB and LC3- 

GFP analysis and mitochondrial respiration by Seahorse assay, 

and prevents temsirolimus-mediated CD133 induction. Pharmacokinetic 

studies reveal orally administered BC is converted to 

~50% less potent baicalin (BCi) with glucuronate substituting 

C7 -OH in plasma but largely reverted to BC in the mouse 

liver. A similar reversion is confirmed in cultured primary 

hepatocytes and TIC treated with BCi. BC treatment completely 

eliminates temsirolimus chemoresistance of TIC-derived subcutaneous 

tumor growth in nude mice. LC/MC analysis of TIC 

proteins pulled-down with BCi-conjugated nanobeads, identifies 

Rab1 and other Rab family members as potential targets of 

BC, and 3D structural analysis and docking studies predicted 

BC interaction at the GTP binding site of Rab1 via BC’s –OH 

of C5 and C7. Indeed, GTP but not ATP competitively inhibits 

BC-Rab1 interaction and Rab1 overexpression rescues BC-induced 

inhibition of autophagy, but not TIC killing, suggesting 

a role for additional Rab protein target(s) for the latter effect. 

[Conclusion] BC promotes killing of chemoresistant TICs, particularly 

when combined with mTORC1 inhibitor via mechanisms 

which appear to involve interactions with Rab family proteins. 

Disclosures: 

Hidekazu Tsukamoto - Consulting: Suntory Ltd.; Grant/Research Support: The 

Toray Co. 

The following authors have nothing to disclose: Raymond Wu, Ramachandran 

Murali, Yasuaki Kabe, Keigo Machida, Clay Wang, Stan G. Louie 

1937 

CD26 (DPP-4) as a molecular target for HCC treatment 

Sohji Nishina 1 , Akira Yamauchi 2 , Yuichi Hara 1 , Keisuke Hino 1 ; 

1 Hepatology and Pancreatology, Kawasaki medical univercity, 

Kurashiki city, Okayama, Japan; 2 Biochemistry, Kawasaki Medical 

School, Kurashiki city, Okayama, Japan 

Background and Aim: CD 26 is a multifunctional transmembrane 

glycoprotein and functions as dipeptidyl peptidase 4 

(DPP-4). Although CD26 is expressed in various cancers, the 

relationship between hepatocellular carcinoma (HCC) progression 

and CD26 expression remains unknown. The aim of this 

study was to investigate the potential role of CD26 as a molecular 

target for HCC treatment. Methods: CD26 expression was 

examined in 41 surgically resected liver specimens from patients 

with HCC. In vitro the effect of DPP-4 inhibitor, anagliptin, on 

cancer cell growth was investigated using Huh7/HepG2 cells 

that expresses CD26. In vivo nude mice (BALBc-nu/nu) were 

subcutaneously injected Huh7 cells and then fed control diet, 

low-dose anagliptin containing diet or high-dose anagliptin 

containing diet for 21 days. Results: CD26 expression was 

correlated with cell proliferation, angiogenesis and cell differentiation 

in HCC specimens obtained from patients. Anagliptin 

did not affect cell proliferation or cell cycle in vitro. However 

in nude mice, anagliptin significantly suppressed the growth of 

xenograft tumors in dose dependent manner. Anagliptin also 

induced NK cells infiltrations more vigorously and reduced 

angiogenesis in xenograft tumors, even though body weight 

and dietary intake through the feeding period, and glucose tolerance 

determined at the 14th day of feeding were not different 

among three groups. These results suggested that anagliptin 

potentially have antitumor activity against HCC. Furthermore, 

we examined whether anagliptin activated chemotaxis of NK


cells, since NK cells were more vigorously infiltrated in mice fed 

the anagliptin containing diet. NK cells obtained human volunteers 

were examined for their chemotaxis using EZ-TAXIScan in 

the presence of CXCL10, chemokine for NK cells, with or without 

anagliptin. Interestingly, anagliptin significantly activated 

the chemotaxis of NK cells probably through the reduction 

CXCL10 antagonism, because it has been shown that CXCL10 

is truncated at its N-terminus through DPP-4 activity and that a 

N-terminal truncated CXCL10 acts as an antagonist form. Conclusions: 

The present results indicated that CD26 expression 

was related to tumor progression in patients with HCC and 

that DPP-4 inhibitor, anagliptin potentially suppressed HCC 

progression through activation of chemotaxis of NK cells, and 

inhibition of angiogenesis. 

Disclosures: 

Akira Yamauchi - Grant/Research Support: Kobayashi Pharmaceutical Co., Ltd. 

The following authors have nothing to disclose: Sohji Nishina, Yuichi Hara, 

Keisuke Hino 

1938 

Liver Selective Acetyl-CoA Carboxylase Inhibitor 

ND-654 Improves Sorafenib Efficacy in the Treatment of 

Hepatocellular Carcinoma in Cirrhotic Rats 

Lan Wei 1 , Omeed Moaven 1 , Geraldine Harriman 2 , Jeremy R. 

Greenwood 3 , Sathesh P. Bhat 3 , William Westlin 2 , H. James Harwood 

2 , Rosana Kapeller 2 , Kenneth K. Tanabe 1 , Bryan C. Fuchs 1 ; 

1 Surgery, Massachusetts General Hospital, Boston, MA; 2 Nimbus 

Therapeutics, Cambridge, MA; 3 Schrodinger, New York, NY 

Background: Hepatocellular carcinoma (HCC) is increasing in 

incidence worldwide. Current treatment options for HCC are 

limited, and as such, prognosis is extremely poor with a 5-year 

survival less than 12%. Sorafenib is the only FDA-approved 

drug for the treatment of HCC but its effects are marginal as it 

only extends survival by a few months. Metabolic attenuation 

is a promising approach to cancer therapy, and rate-limiting 

steps in key biosynthetic pathways are particularly attractive 

targets. We recently identified ND-654, a hepatoselective 

(~3000:1 liver to muscle exposure), allosteric ACC inhibitor 

that binds to the ACC subunit dimerization site and inhibits the 

enzymatic activity of both ACC1 (IC50 = 3 nM) and ACC2 

(IC50 = 8 nM). We previously demonstrated that daily oral 

administration of 10 mg/kg ND-654 reduced tumor incidence 

by 55% and significantly improved median survival time in a 

rat model of cirrhosis and HCC. Here, we examine the effects 

of ND-654 alone and in combination with sorafenib on HCC 

development in cirrhotic rats. Methods: Male Wistar rats were 

treated once weekly with 50 mg/kg diethylnitrosamine (DEN) 

for 18 weeks to induce sequential development of fibrosis, 

cirrhosis and HCC. After establishment of cirrhosis and when 

HCCs are first developing (13 weeks), rats were treated daily 

by oral gavage with either 1) vehicle control, 2) 10 mg/kg 

ND-654, 3) 10 mg/kg sorafenib, or 4) 10 mg/kg ND-654 

and 10 mg/kg sorafenib. After 18 weeks, tumor nodules 

were counted and liver and tumor tissue was harvested for 

analysis. Results: Similar to our previous study, simultaneous 

inhibition of ACC1 and ACC2 significantly reduced HCC incidence 

by 41% (p < 0.05) which was comparable to results 

with sorafenib (57% reduction, p < 0.01). Remarkably, the 

combination of ND-654 and sorafenib significantly reduced 

HCC incidence by 81% (p < 0.001). Our previous results had 

shown that ND-654 decreased tumor proliferation and induced 

necrosis and we are currently examining the combination of 

ND-654 and sorafenib on these endpoints. Conclusions: These 

results provide further evidence that de novo lipogenesis is an 

important mediator of hepatic carcinogenesis and that selective 

inhibition of hepatic ACC is a viable cancer metabolism 

therapeutic strategy for treating HCC that could be combined 

with sorafenib. 

Disclosures: 

Jeremy R. Greenwood - Employment: Schrodinger Inc.; Patent Held/Filed: Nimbus 

Therapeutics; Stock Shareholder: Schrodinger Inc. 

Rosana Kapeller - Employment: Nimbus Therapeutics; Stock Shareholder: Aileron 

Therapeutics 

Kenneth K. Tanabe - Patent Held/Filed: EGF SNP and risk for HCC, EGFR inhibition 

and HCC, gene signature for prognosis in cirrhosis 

Bryan C. Fuchs - Consulting: Collagen Medical; Grant/Research Support: Nimbus 

Therapeutics 

The following authors have nothing to disclose: Lan Wei, Omeed Moaven, Geraldine 

Harriman, Sathesh P. Bhat, William Westlin, H. James Harwood 

1939 

The Long Noncoding RNA SNHG6 Functions As a Competing 

Endogenous RNA to Promote Hepatocellular Carcinoma 

Progression 

LI LIU 1 , Chuanhui Cao 2 , Ting Ting Zhang 1 , Dongyan Zhang 2 , 

Dehua Wu 2 ; 1 Institution of hepatology, Guangzhou, China; 

2 Department of Radiation Oncology, Guangzhou, China 

Abstract: Expression of long noncoding RNAs (lncRNAs) has 

been reported to be dysregulated in hepatocellular carcinoma 

(HCC) but their contributions and functional mechanisms to 

HCC remain largely unknown. Here, we elucidate whether 

lncRNA SNHG6 is involved in HCC progression.Our results 

showed that SNHG6 was up-regulated in HCC in 160 paired 

cancerous and noncancerous tissues by quantitative real-time 

PCR and in situ hybridization. SNHG6 associated with portal 

vein tumor thrombus, tumor stage, metastasis, and shorter overall 

survival of HCC patients. Ectopic expression of the SNHG6 

in HCC cells promoted their proliferation and inhibited apoptosis 

both in vitro and in vivo. Furthermore, we revealed an 

endogenous interaction between SNHG6 and miR-26a/b by 

coimmunoprecipitation with anti-Ago2 in HCC cells. More 

importantly, SNHG6 controlled the expression of miR-26a/b 

target gene, TAK1. In our in vitro and in vivo system, and 

clinical HCC tissues, we observed that SNHG6 positively correlated 

with TAK1 and ectopic expression of SNHG6 was sufficient 

to increase TAK1. Moreover, knockdown TAK1 could 

arrest HCC cell proliferation inducing by overexpression of 

SNHG6, which was consistent with results of knockdown of 

SNHG6 expression in HCC cells. Notably, there also exists 

co-expression relationship between SNHG6 and TAK1 in liver, 

colon, rectum, and pancreas cancers.CONCLUSIONS: SNHG6 

may act as a biomarker of poor prognosis in HCC and confer 

malignant phenotype to tumor cells. The ceRNA network involving 

SNHG6 may contribute to a better understanding of liver 

carcinogenesis and tumor progression.


follow-up, association of OPN mRNA expression in liver or in 

tumor tissue with pattern of HCC recurrence was determined. 

Male wild-type (WT) littermates, Opn global knockout (Opn -/- ) 

and transgenic mice overexpressing Opn in hepatocytes in 

either WT (Opn Hep Tg) or in Opn -/- background (Opn -/-Hep Tg) 

were injected diethylnitrosamine (DEN) at 15 d of age and 

were followed for the development of liver tumors up to 1 yr. 

Upon sacrifice, histopathological scoring, immunohistochemistry 

for OPN, ELISA for serum and liver α-fetoprotein (AFP) and 

serum transaminases were analyzed. RESULTS: Elevated OPN 

mRNA in non-tumor liver tissue from 199 HBV-related HCC 

patients was associated with late tumor recurrence (>2 yrs 

after surgery, de novo carcinogenesis from remnant diseased 

liver) but not with early recurrence (dissemination/metastasis 

of resected primary tumor). The same trend was not observed 

in a second cohort of patients with HCV-related HCC thus suggesting 

HBV specificity. In contrast, OPN mRNA in tumor tissue 

correlated with early but not with late recurrence, indicating a 

dual oncogenic role of OPN according to the site of expression. 

To understand the mechanism whereby intrahepatic OPN 

and specifically hepatocyte OPN contributed to HCC, a wellknown 

mouse model mimicking HCC was used. After 1 yr 

of DEN injection, Opn -/- mice exhibited significantly reduced 

macroscopic HCC development, less number of HCC nodules 

per sample and lower serum and hepatic AFP levels compared 

to WT littermates. Yet, overexpression of Opn in hepatocytes 

rescued the phenotype from Opn -/- mice suggesting a key role 

for hepatocyte-derived OPN in HCC development, which was 

further confirmed by the leading incidence of tumors, the largest 

tumor size and the highest AFP in Opn Hep Tg mice. CON- 

CLUSIONS: 1) Liver and more importantly hepatocyte-derived 

OPN is a major player orchestrating the development of HCC; 

2) OPN specifically plays a role in HBV-related HCC and 3) 

Ablation or reduction of hepatocyte-derived OPN may be a 

useful strategy to prevent the onset and/or the progression to 

HCC. 

Disclosures: 

The following authors have nothing to disclose: Yu Chen, Xiaodong Ge, Ioana 

Abraham-Enachescu, Xiaochen Sun, Grace Guzman, Yujin Hoshida, Natalia 

Nieto 

Disclosures: 

The following authors have nothing to disclose: LI LIU, Chuanhui Cao, Ting Ting 

Zhang, Dongyan Zhang, Dehua Wu 

1940 

Hepatocyte-derived osteopontin promotes the development 


Yu Chen 1 , Xiaodong Ge 1,2 , Ioana Abraham-Enachescu 2 , Xiaochen 

Sun 2 , Grace Guzman 1 , Yujin Hoshida 2 , Natalia Nieto 1,2 ; 1 Pathology, 

University of Illinois at Chicago, Chicago, IL; 2 Department of 

Medicine, Icahn School of Medicine at Mount Sinai, New York, 

NY 

BACKGROUND & HYPOTHESIS: Previous work from our laboratory 

demonstrated that intrahepatic osteopontin (OPN) is highly 

profibrogenic. Since fibrosis progression is a major cause of 

hepatocellular carcinoma (HCC) and the role of OPN in the 

pathogenesis of HCC is unknown, we hypothesized that the 

presence of OPN in the liver microenvironment and specifically 

in hepatocytes could promote the development of HCC. METH- 

ODS: To prove this hypothesis, we analyzed clinical cohorts of 

HCC patients and a mouse model of HCC with genetic manipulation 

of Opn. In HCC patients’ cohorts with long-term clinical 

1941 

Efficacy and mechanism of dendritic cell-based immunotherapy 

in combination with T cell immunoglobulin and 

mucin protein 3 blockade in a murine hepatocellular 

carcinoma model 

Junichi Eguchi 1 , Eiichi Hayashi 2 , Hiroyoshi Doi 2 , Risa Omori 2 , 

Atsushi Kajiwara 2 , Hitoshi Yoshida 2 , Takayoshi Ito 1 , Yoshio Deguchi 

1 , Norihiro Nomura 1 , Haruhiro Inoue 1 ; 1 Digestive Disease 

Center, Showa University Koto-Toyosu Hospital, Tokyo, Japan; 

2 Division of Gastroenterology, Department of Medicine, Showa 

University School of Medicine, Tokyo, Japan 

Progress in treatments for hepatocellular carcinoma (HCC) has 

improved the prognosis of patients with HCC. However, HCC 

is usually associated with cirrhosis and often recurs even after 

complete treatment of the tumors in the remaining part of the 

cirrhotic liver. Thus, there is a strong need for the development 

of a new intervention therapy that suppresses the occurrence or 

recurrence effectively with fewer side effects. Immunotherapy 

may be such a treatment and several clinical trials have been 

performed for HCC treatment. Dendritic cells (DCs) are known 

as professional antigen-presenting cells characterized by their 

potent ability to elicit immune responses to tumor-specific T 

cells against tumor-associated antigens. T cell immunoglobulin 

and mucin protein 3 (TIM-3), has been identified as a marker


of immunosuppressive statement. Interaction of TIM-3 with its 

ligand, galectin 9, triggers cell death in activated T cells. In this 

study, we evaluated the efficacy of the combination of DC-based 

vaccine and TIM-3 blockade, and investigated the mechanisms 

of the antitumor effects of the combined therapy. In protection 

model, mice were injected with DCs or/and TIM-3-antibody 

before the murine HCC tumor cell (BNL cell) challenge. 50% of 

mice treated with both DCs and TIM-3-antibody rejected tumor 

challenge, whereas other groups observed a palpable tumor in 

all mice tested. In therapeutic model, tumor-bearing mice were 

inoculated with DCs or/and TIM-3-antibody. Significant suppression 

of outgrowth of the established tumors was observed 

in the DCs and anti-TIM-3 combination treatment group (DCs + 

anti-TIM-3, 265.3 ± 53.09 mm 2 vs control, 535.1 ± 44.47 mm 2 

on day 30, P = 0.0046 vs controls). To investigate induction of 

tumor-specific immune responses, we stimulated splenocytes of 

DCs or/and TIM-3-antibody treated mice twice weekly by DCs 

in vitro. High tumor-specific immune response was detected 

when splenocytes of mice treated with both DCs and anti-TIM-3 

were used as effector cells in interferon-gamma enzyme-linked 

immunospot assay. In addition, incubation with TIM-3-antibody 

enhanced expression of IL-12p70, and cell surface expression 

of CD80 in bone marrow-derived DCs. Our findings suggest 

that blockade of the TIM-3 enhanced the maturation of DCs, 

and Th1-type antitumor immune responses induced by DC vaccine. 

The combination of DC vaccine and TIM-3 blockade may 

be a possible candidate for a cancer vaccine for clinical trials. 

Disclosures: 

The following authors have nothing to disclose: Junichi Eguchi, Eiichi Hayashi, 

Hiroyoshi Doi, Risa Omori, Atsushi Kajiwara, Hitoshi Yoshida, Takayoshi Ito, 

Yoshio Deguchi, Norihiro Nomura, Haruhiro Inoue 

1942 

PTEN Expression Affects the Outcome of Human Hepatocellular 

Carcinoma via Modulating the Oncogenic 

Behaviors and the angiogenic Processes 

Tsung-Hui Hu 1 , Po-Lin Tseng 1 , Pey-Ru Lin 1 , Ming-Hong Tai 2 , Chao- 

Cheng Huang 3 , Li-Na Yi 1 , Chih-Chi Wang 4 , Kuo-Chin Chang 1 , 

Yi-Hao Yen 1 , Ming-Chao Tsai 1 , Ming-Tzung Lin 1 ; 1 Division of 

Hepato-Gastroenterology, Department of Internal Medicine,, 

Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan; 

2 Institute of Biological Sciences, National Sun Yat-Sen University, 

Kaohsiung, Taiwan; 3 Department of Pathology, Kaohsiung Chang 

Gung Memorial Hospital, Kaohsiung, Taiwan; 4 Department of 

Surgery, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, 

Taiwan 

Reduced phosphatase and tensin homolog (PTEN) expression 

is associated with vascular endothelial growth factor (VEGF) 

overexpression in various malignancies. Herein, we aimed to 

investigate the expression of PTEN and VEGF in pericancerous 

tissues and their association in hepatocellular carcinoma 

(HCC). PTEN expression was reduced in 43% and VEGF was 

overexpressed in 31% of 113 resected HCC clinical specimens. 

VEGF overexpression is positively correlated with young 

age, male gender, hepatitis B viremia, high α-fetoprotein 

levels, decline in PTEN expression, advanced tumor stage, 

and dedifferentiation of HCC. Survival analysis revealed that 

reduced PTEN expression, microvessel density, and advanced 

tumor stage were independent poor prognostic factors for disease-free 

survival. Adenovirus-mediated restoration of PTEN 

inhibited cell proliferation, clonogenic growth and invasive 

capability of PTEN-deficient human HCC cells in vitro. In xenograft 

model, PTEN gene delivery could efficiently suppress the 

incidence and growth rate of implanted tumor cells and reduce 

microvessel density. The expression of VEGF and HIF1a, and 

secreted IL-8 were decreased in PTEN restored HCC cells. 

Angiogenic networks of human umbilical vein endothelial cells 

were also suppressed by the increased PTEN in vitro. Conclusion: 

The tumor microenvironment may be affected by PTEN 

expression through modulating the oncogenic phenotypes of 

tumor cells and having a tendency to interfere in angiogenic 

processes, including expression of angiogenic factors and formation 

of angiogenic networks. The expression of PTEN and 

VEGF affecting the outcomes of HCC patients might serve as 

good prognostic biomarkers. 

Disclosures: 

The following authors have nothing to disclose: Tsung-Hui Hu, Po-Lin Tseng, 

Pey-Ru Lin, Ming-Hong Tai, Chao-Cheng Huang, Li-Na Yi, Chih-Chi Wang, Kuo- 

Chin Chang, Yi-Hao Yen, Ming-Chao Tsai, Ming-Tzung Lin 

1943 

Inhibition of autophagy reverses the resistance to 

sorafenib induced by hypoxia in hepatocellular carcinoma 

Takayuki Kogure, Yasuteru Kondo, Jun Inoue, Yu Nakagome, Yuta 

Wakui, Tatsuki Morosawa, Yasuyuki Fujisaka, Teruyuki Umetsu, 

Tooru Shimosegawa; Tohoku University Hospital, Sendai, Japan 

Backgrounds: Sorafenib is the only effective systemic therapy 

for hepatocellular carcinoma (HCC). It shows cytotoxicity on 

HCC cells in vitro but its clinical response is disease stabilization. 

Sorafenib shows inhibition of angiogenesis, which could 

lead the tumor microenvironment to severe hypoxia. It has been 

reported that hypoxia is closely related to resistance to many 

types of anticancer drugs. We hypothesized that hypoxia 

induces resistance to sorafenib through activation of autophagy, 

which results in clinically modest efficacy of sorafenib. 

We investigated the involvement of autophagy in the resistance 

to sorafenib in HCC. Methods: HCC cell lines, PLC/PRF/5 

(PLC5), Li7, HepG2, and Huh7 were cultured in 95% air and 

5% CO 2 

(normoxia). For hypoxia, cells were transferred to a 

hypoxia chamber with 1% O 2 

, 5% CO 2 

, and 94% N 2 

. Proliferation 

of HCC cells was assessed with growth curve assay 

and MTS assay. Apoptosis was assessed morphologically and 

with activation of caspase-3/7 using a luminometric assay. 

Hypoxia inducible factor 1α (HIF1α) protein was detected by 

immunoblotting. Involvement of autophagy was assessed by 

detecting LC3 and p62 and by treating cells with chloroquine 

(CQ), an inhibitor of autophagy. Results: After 24 hr of cell 

culture in hypoxia, PLC5 and Li7 showed significant increase of 

HIF1α but no significant change was observed in growth curve 

through 96 hr of culture. Sorafenib inhibited cell growth in a 

concentration-dependent manner and this effect was reduced 

under hypoxic condition with an IC 50 

(normoxia/hypoxia, μM) 

of 5.9/9.4 in PLC5, 7.4/11.4 in Li7, 12.1/14.5 in Huh7, and 

6.9/11.7 in HepG2. Induction of apoptosis was suppressed 

under hypoxia. Incubation with sorafenib (5 and 10 μM) for 

24 hr induced apoptosis by 14.6% and 23.8% in PLC5 in normoxia 

whereas 10.5% and 13.1% in hypoxia. An increase of 

caspse-3/7 activity induced by sorafenib (5 and 10 μM) was 

reduced by 77.2% and 55.3% under hypoxia. To assess the 

involvement of autophagy, protein expression of LC3 and p62 

was detected. After 48 hr of culture in hypoxia, an increase of 

LC3-II and a decrease of p62 was observed in PLC5 and Li7, 

indicating the activation of autophagy. HCC cells were treated 

with sorafenib with or without CQ. Under hypoxia, viability 

of cells treated with sorafenib (10 μM, 72 hr) was 55.6% in 

PLC5 and 72.1% in Li7. Co-treatment of cells with CQ (50 μM) 

recovered the cytotoxicity of sorafenib with a viability of 86.4% 

in PLC5 and 89.9% in Li7. Conclusion: Under hypoxia, resistance 

to sorafenib is induced by the activation of autophagy.


Inhibition of autophagy could be an attractive approach for 

potentiating the antitumor effect of sorafenib in HCC. 

Disclosures: 

The following authors have nothing to disclose: Takayuki Kogure, Yasuteru 

Kondo, Jun Inoue, Yu Nakagome, Yuta Wakui, Tatsuki Morosawa, Yasuyuki 

Fujisaka, Teruyuki Umetsu, Tooru Shimosegawa 

1944 

The relationship between hepatocellular carcinoma 

development and reduced expression levels of the 

major germline genotype of PROSC, which is frequently 

deleted in hepatocellular carcinoma 










Background & Aims: In this study, we focused on the recurrently 

amplified or deleted genes in hepatocellular carcinoma (HCC) 

tissues and examined the association between their baseline 

expression levels, which are regulated by single nucleotide 

polymorphisms (SNPs), and development of HCC. Methods: 

We searched for expression quantitative trait loci (eQTLs) for 

previously reported genes with copy number alterations from 

a public database of non-transformed peripheral blood samples 

that includes over 5,000 individuals. We then conducted 

a case-control analysis of these eQTL SNPs or SNPs linked 

to them (r 2 > 0.8 in both European and Asian populations) 

using our SNP typing data in HCC cases (631 HBV-related 

HCC, 554 HCV-related HCC) and 5,489 controls without 

any liver diseases or cancers. Results: Among 13 recurrently 

amplified genes, we found 10 cis-acting eQTL SNPs (i.e., the 

SNP is located near the expressed gene) and 2 trans-acting 

eQTL SNPs (i.e., the SNP is located far from the expressed 

gene or on another chromosome). Among 18 recurrently 

deleted genes, we found 15 eQTL SNPs and 3 trans-acting 

eQTL SNPs. Among these 30 eQTL SNPs, we have successfully 

genotyped 22 of either the target SNP, when available, or 

else a closely linked SNP. We genotyped 9 cis-acting and 1 

trans-acting eQTL SNPs for amplified genes, and 9 cis-acting 

and 3 trans-acting eQTL SNPs for deleted genes. In case-control 

analysis, no significant association was observed between 

SNP frequencies and HCC development both in HBV-related 

and HCV-related HCC studies (vs. controls). Subsequently, we 

compared each HCC case to chronic HBV and HCV patients 

without HCC (n = 1,997 and 1,354). After adjustment for 

multiple testing, we found 2 significantly associated eQTL SNPs 

with HCC for PROSC in the HBV study (Odds ratio [OR] = 

1.27) and for TNFAIP3 in the HCV study (OR = 1.37), suggesting 

their baseline expression levels may be important for 

virus-related hepatocarcinogenesis. The risk allele of the eQTL 

SNP for HBV-related HCC strongly down-regulates PROSC (P 

= 8.25 × 10 -149 ), whereas the risk allele of the eQTL SNP 

for HCV-related HCC weakly up-regulates TNFAIP3 (P = 2.60 

× 10 -4 ). Furthermore, both PROSC and TNFAIP3 are recurrently 

deleted genes in HCC. Taken together, the association 

of the eQTL SNP for PROSC with HBV-related HCC susceptibility 

seems to be more compatible than that for TNFAIP3 with 

HCV-related HCC susceptibility. Conclusions: As a result of 

integrative analysis using our large-scale genome-wide SNP 

genotype data and public eQTL data, we suggest that germline 

variants of recurrently deleted genes may affect viral hepatocarcinogenesis 

by regulating their expression levels. 

Disclosures: 













1945 

Steatosis Induces Wnt/Β-Catenin Pathway To Stimulate 

Proliferation Of Hepatic Tumor Initiating Cells And Promote 

Liver Cancer Development 

Anketse D. Kassa 1 , Ni Zeng 1 , Vivian G. Medina 1 , Lina He 1 , Kaiting 

Yang 1 , Indra Mahajan 1 , Chien-Yu Chen 1 , Richa Aggarwal 1 , 

Zhechu Peng 1 , Megan Rieger 2 , Chia-Lin Chen 2 , Keigo Machida 2 , 

Zea Borok 2 , Michael Kahn 2 , Bangyan L. Stiles 1,2 ; 1 Pharmacology 

and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, 

CA; 2 Keck School of Medicine, USC, Los Angeles, CA 

Obesity is a major compounding factor for liver cancer where 

male with a body mass index (BMI) >35 has 4.52 fold higher 

incidence of liver cancer vs. those with BMI between 18.5 and 

24.9. In mouse models, constitutive activation of the insulin/ 

PI3K signaling pathway by deletion of the negative regulator 

Pten (phosphatase and tensin homologue deleted on chromosome 

10) leads to hepatosteatosis and later tumor phenotypes. 

In this model (Pten loxP/loxP ; Alb-Cre + ), we have shown that the 

presence of the underlying fatty liver condition is required for 

the development of liver cancers, which expands from a population 

of tumor initiating cells (TICs). In this study, we investigated 

how fatty liver disease may contribute to cancer formation. 

We show here that blocking fatty liver formation by genetic 

or non-genetic approaches significantly reduced the TIC population. 

Concomitantly, tumor development in the Pten loxP/loxP ; 

Alb-Cre + mice is inhibited despite of the tumor transformation 

signals of PTEN loss. We further demonstrated that hepatic 

steatosis resulting from Pten deletion or high fat diet feeding 

induces the activation of Wnt/b-catenin pathway. Genomic 

profiling of human liver cancer specimens has indicated a high 

prevalence of b-catenin activating mutation. However, consistent 

with mouse studies that genetically modify b-catenin, 

such mutation is not sufficient to drive malignant transformation. 

We show here that Wnt/b-catenin activation signal is the 

tumor promoting factor induced by lipid accumulation and that 

this signal is critical for the expansion of the transformed liver 

TICs. In the Pten loxP/loxP ; Alb-Cre + mice fed low caloric diet to 

inhibit hepatic steatosis, reduced expression of Wnt ligands 

is observed concurrent with inhibited expansion of TICs and 

complete blockage of tumor development. Blocking the signal 

of Wnt/b-catenin with shRNA or a pharmacological reagent 

ICG-001 inhibits the proliferation of TICs in vitro and lead to 

decreased accumulation of TICs and reduction of tumor grafts 

in vivo. Together, our study suggests that fatty liver activates the 

niche factor Wnt/b-catenin to promote tumorigenesis using the 

Pten null liver cancer model. 

Disclosures: 

The following authors have nothing to disclose: Anketse D. Kassa, Ni Zeng, Vivian 

G. Medina, Lina He, Kai-ting Yang, Indra Mahajan, Chien-Yu Chen, Richa 

Aggarwal, Zhechu Peng, Megan Rieger, Chia-Lin Chen, Keigo Machida, Zea 

Borok, Michael Kahn, Bangyan L. Stiles


1946 

Truncated Hepatitis B virus X protein up-regulates 

CD44+ cancer stem like cells through LTBP1 

Goki Suda, Seiji Tsunematsu, Masato Nakai, Jun Itoh, Mitsuteru 

Natsuizaka, Kenichi Morikawa, Koji Ogawa, Naoya Sakamoto; 

hokkaido university, Sapporo, Japan 

Background Aims: Hepatitis B virus (HBV) is a major pathogen 

of hepatocellular carcinoma (HCC). HBV X protein (HB) is 

reported to promote HCC development by activation of various 

signals. Recently genome-wide association study about HBV-related 

HCC revealed that COOH-truncated HBx was integrated 

in host genome at high frequency. Cancer stem like cells (CSCs) 

are defined as those cells that can self-renew, drive tumorigenesis. 

It is not clear whether COOH-truncated HBx integration 

promotes prevalence of CSCs or not. Therefore, we aimed 

to analyze the relationship between COOH-truncated HBx 

expression and CSCs. Methods: We firstly established stably 

HBx (HBx/HepG2) and COOH-truncated HBx (truncated HB/ 

HepG2). Next we examined the expression levels of stemness 

makers in these established cells by Flow cytometry analysis. 

Subsequently we analyzed the anti-tumor agent resistance and 

tumorigenesis of these cells by MTS assay and soft gel agar 

colony formation assay. Finally we investigated the factors that 

promote prevalence of CSCs in truncated-HBx /HepG2 cells 

by PCR-array. Results: Both HBx/HepG2 and truncated HBx/ 

HepG2 shows significantly resistant to anti-tumor agent, 5-FU, 

compared with control/HepG2 cells. Soft gel agar colony formation 

assay shows that truncated-HBx/HepG2 cells make significantly 

more colonies than control/HepG2 cells and HBx/ 

HepG2 cells. Flow cytometry analysis reveals that truncated 

HBx/HepG2 cells express high levels of CD44, one of cancer 

stemness maker, compared with control/HepG2 and HBx/ 

HepG2 cells. Next truncated-HBx cells were sorted in CD44 

high and low cells and were analyze potential of anti-tumor 

agent resistant and tumorigensis. CD44high cells shows resistant 

to anti-tumor agent 5-FU compared with CD44low cells, 

and soft gel agar colony formation assay shows that CD44high 

cells make significantly higher colonies than CD44low cells. 

Subsequently we investigated what factor up-regulate CD44 in 

truncated HBx/HepG2 cells by Human Stem Cell PCR Array 

(Qiagen).This PCR-array results indicated that LTBP1 is significantly 

up-regulated in only Truncated-HBx/HepG2 compared 

with HBx/HepG2 and con/HepG2 cells. The expression level 

of CD44 were significantly down-regulated by siRNA knockdown 

of LTBP1. In sorted CD44high cells, LTBP1 were significantly 

up-regulated compared with CD44low cells. Conclusion: 

COOH-truncated HBx protein up-regulates CD44+ cancer stem 

like cells via LTBP1. This finding might contribute development 

of efficient strategies for eliminating CSCs 

Disclosures: 



The following authors have nothing to disclose: Goki Suda, Seiji Tsunematsu, 

Masato Nakai, Jun Itoh, Mitsuteru Natsuizaka, Kenichi Morikawa, Koji Ogawa 

1947 

P300 promotes TGF-β stimulated nuclear translocation 

of SMADs and activation of hepatic stellate cells into 

liver metastasis promoting myofibroblasts 

Luyang Guo 2 , Xiaoyu Xiang 2 , Vijay Shah 1 , Ningling Kang 2 ; 

1 Mayo Clinic, Rochester, MN; 2 the Hormel Institute, Austin, MN 

Introduction: Liver metastasis is dependent on bidirectional interactions 

between cancer cells and the microenvironment of the 

liver. TGF-β, released from cancer cells and other cells within 

the liver, induces activation of hepatic stellate cells (HSCs) into 

myofibroblasts (MFs). In turn, MF/activated HSCs promote liver 

metastatic growth. TGF-β induces intracellular signaling events 

in HSCs, including activation of receptors, nuclear translocation 

of SMADs and transcription of TGF-β target genes, which 

present targets to inhibit HSC activation. However, mechanisms 

governing these signaling events remain poorly understood. 

Hypothesis: We tested if p300, an acetyltransferase 

previously known to promote gene transcription, modulates 

TGF-β1 induced activation of HSCs into tumor promoting MFs. 

Methods: Lentiviruses encoding different p300 shRNAs were 

used to knockdown p300 and a small molecule C646 was 

used to inhibit p300 acetyltransferase activity. A tumor/HSC 

coculture and coimplantation mouse model were used to test 

the role of p300 knockdown HSCs for tumor growth. Immunofluorescence 

(IF) was used to study subcellular localization of 

p300 and SMAD nuclear translocation. Western Blot analysis 

(WB) and IF for α-SMA were used to assess HSC activation, 

co-immunoprecipitation (coIP) was used to study protein-protein 

interactions and microarray analysis was used to study 

gene expression profiles of HSCs. Results: p300 knockdown 

HSCs were less effective on promoting tumor cell proliferation 

than control HSCs in a tumor/HSC coculture and were less 

effective on promoting tumor growth in mice in a subcutaneous 

tumor/HSC coimplantation model. In vitro, p300 knockdown 

inhibited TGF-β1 stimulated activation of HSCs into MFs as 

assessed by both WB and IF for α-SMA (p200 cells 

per group). IF revealed that p300 resided in both cytoplasm 

and nucleus of HSCs and that p300 knockdown or inhibition 

by C646 inhibited TGF-β1 stimulated nuclear translocation of 

SMADs. Co-IP demonstrated that TGF-β1 stimulation promoted 

a p300/Importin7/8/SMADs complex in the cytoplasm that 

promotes nuclear transport of SMADs. Furthermore, microarray 

analysis identified that p300 knockdown suppressed transcription 

of TGF-β1 inducible genes, such as CTGF, TNC, PDGFC, 

FGF2 and POSTN encoding either prometastatic niche factors 

or growth factors of tumor cells. Conclusions: p300 is required 

for TGF-β1 stimulated nuclear translocation of SMADs and transcription 

of TGF-β1 target genes encoding numerous tumor 

promoting paracrine factors. p300 of HSCs thus presents a 

therapeutic target to inhibit HSC activation required for the 

development and progression of liver metastasis. 

Disclosures: 

The following authors have nothing to disclose: Luyang Guo, Xiaoyu Xiang, Vijay 

Shah, Ningling Kang 

1948 

Critical role of Hippo cascade in regulating AKT/Rasdriven 

liver cancer development in mice 

Shanshan Zhang 2,1 , Junyan Tao 1 , Xiaolei Li 1 , Diego Calvisi 3 , 

Xin Chen 1 ; 1 Department of Bioengineering and Therapeutic Sciences, 

University of California San Francisco, San Francisco, CA; 

2 Department of Pathology and Pathophysiology, Eastern Hepatobiliary 

Surgery Hospital, Second Military Medical University, Shanghai, 

China; 3 Institut für Pathologie, Ernst-Moritz-Arndt-Universität, 

Greifswald, Germany 

Background: Primary liver cancer consists of both hepatocellular 

carcinoma (HCC) and intrahepatic cholangiocarcinoma 

(ICC). Hippo pathway is a critical regulator in liver cancer 

development. However, the precise role of the Hippo cascade 

along hepatocarcinogenesis has not been fully explored. Purpose: 

In this study, we investigated the functional significance 

of Yap and TAZ, the transcriptional activators downstream of 

Hippo tumor suppressor kinases, in a murine model of liver 

cancer. Methods: We employed a mixed HCC and ICC mouse 

model via hydrodynamic transfection of activated forms of AKT


and Ras oncogenes (AKT/Ras), based on the evidence that activation 

of AKT/mTOR and Ras/MAPK pathways often occurs in 

human HCC and ICC samples. In AKT/Ras mice, either the 

Hippo cascade was activated by co-injecting Lats2 or Yap/ 

TAZ activity was blocked via transfection of dominant negative 

TEAD2 (dnTEAD2). Furthermore, Yap or TAZ was silenced via 

mir30 based shRNA. qRT-PCR, Western blotting, and immunohistochemistry 

(IHC) were applied to detect gene and protein 

expression patterns in tissues and cell lines. Results: Both Yap 

and TAZ showed nuclear staining in AKT/Ras liver tumor samples. 

Co-injecting Lats2 or dnTEAD2 with AKT/Ras strongly 

delayed liver tumor development. Similar results were obtained 

when the co-expression of AKT/Ras was paralleled by Yap or 

TAZ silencing in the mouse liver. Histological analysis showed 

decrease in liver tumor number and tumor size as well as the 

prevalence of adenomas over carcinomas in the liver of AKT/ 

Ras mice. Importantly, activation of Hippo or loss of Yap/TAZ 

led to the complete elimination of ICC-like lesions in the liver. 

The phenotype was highly similar to that observed when AKT/ 

Ras mice were treated with anti-Notch2 antibodies. Indeed, 

further analysis demonstrated the down-regulation of Notch2 

in these mouse liver tumor tissues. Silencing of Yap or TAZ in 

human HCC cell lines also led to the decreased expression of 

CCA-like markers such as Sox9 and CK19. Conclusion: Our 

data demonstrate that Hippo pathway negatively regulates 

liver tumor formation in mice via regulating Notch2. Both Yap 

and TAZ are required for hepatocarcinogenesis. 

Disclosures: 

The following authors have nothing to disclose: Shanshan Zhang, Junyan Tao, 

Xiaolei Li, Diego Calvisi, Xin Chen 

1949 

Hepatocyte-specific overexpression of B cell leukemia-3 

in mice suppresses chemically-induced hepatocarcinogenesis 

Nadine Gehrke 1 , Marcus A. Woerns 1 , Yvonne Alt 1 , Nadine Hoevelmeyer 

2 , Ari Waisman 2 , Peter R. Galle 1 , Jörn M. Schattenberg 1 ; 

1 I. Department of Medicine, University Medical Center of the 

Johannes Gutenberg University Mainz, Mainz, Germany; 2 Institute 

for Molecular Medicine Mainz, University Medical Center, Mainz, 

Germany 

Background: B cell leukemia-3 (Bcl-3), which was originally 

described as a proto-oncogene in B cell lymphomas, is highly 

expressed in the liver. It binds tightly to the NFkB subunits 

p50 and p52 homodimers in order to activate or inhibit transcription 

from NFkB-dependent promoters regulating especially 

inflammation and cell death. Bcl-3, when dysregulated, has be 

shown to be widely expressed in several cancer types including 

hepatocellular carcinoma (HCC), but a complete understanding 

of the function of Bcl-3 in cancer development is still 

lacking. Methods: To evaluate the role of hepatic Bcl-3 in hepatocarcinogenesis 

we employed the two-step diethylnitrosamine 

(DEN)/phenobarbital (PB)-liver cancer mouse model. 7-10 

day-old, male mice exhibiting an increased hepatocyte-specific 

expression of Bcl-3 (alfp-Cre:bcl-3, Bcl-3 hepar mice) and wild 

type (wt) littermates received a single intraperitoneal injection 

of 25mg DEN followed, 4 weeks later, by continuous treatment 

with 0.5g/l PB dissolved in drinking water. Blood and liver 

tissue were harvested at 40 weeks of age for further analysis. 

Results: Remarkably, Bcl-3 hepar mice exhibited less tumours compared 

to wt mice in response to DEN/PB. Also their relative 

liver weight was significantly lower, which could be attributed 

to the smaller tumour number and size. Histopathological 

tumour grading revealed a comparable presence of dysplastic 

foci and nodules in all H&E stained liver sections irrespective 

of the genotype. However, more HCC as well as a greater 

relative HCC area were detectable in the wt compared to the 

Bcl-3 hepar group. In addition, K i 

-67 labeling and appropriate 

qRT-PCR analyses (K i 

-67, p53, cMyc, mTOR) showed significantly 

diminished levels of cell proliferation in the HCC region 

of Bcl-3 hepar livers compared to wt livers. Moreover, DEN/PB 

treatment resulted in a markedly reduced liver injury in Bcl- 

3 hepar mice relative to the wt as demonstrated by lower serum 

ALT and LDH levels. In agreement, JNK and ERK activation 

were more pronounced in tumour and residual tumour tissues of 

wt mice, whereas a higher grade of phosphorylated p38 and 

NFkB p65 was determined in Bcl-3 hepar livers protecting against 

DEN/PB-induced cell death. Compared to the wt, the absolute 

number and activation of intrahepatic B cells, that exert a suppressive 

function on tumour growth, was reduced in Bcl-3 hepar 

mice as measured by FACS analysis and decreased mRNA 

expression of CD81 and BLNK. Conclusion: Overexpression of 

hepatic Bcl-3 impaired DEN/PB-induced hepatocarcinogenesis 

through regulation of cell death and inflammatory processes. 

Thus, Bcl-3 could be an important target in the development of 

novel therapies. 

Disclosures: 

Marcus A. Woerns - Advisory Committees or Review Panels: Bayer, Bayer 

Peter R. Galle - Advisory Committees or Review Panels: Bayer, BMS, Lilly, Daiichi, 

Jennerex; Consulting: Medimmune; Grant/Research Support: Roche, Lilly; Speaking 

and Teaching: Bayer, BMS 

The following authors have nothing to disclose: Nadine Gehrke, Yvonne Alt, 

Nadine Hoevelmeyer, Ari Waisman, Jörn M. Schattenberg 

1950 

Platelet-Derived Growth Factor D (PDGF-D)-Induced 

Secretion of Vascular Endothelial Growth Factor-C by 

Cancer-Associated Fibroblasts (CAF) Stimulates Lymphangiogenesis 

in Cholangiocarcinoma 

Massimiliano Cadamuro 2,1 , Marta Vismara 2 , Simone Brivio 2 , 

Mario Strazzabosco 2,3 , Luca Fabris 1,3 ; 1 Dep. of Molecular Medicine, 

University of Padova, Padova, Italy; 2 Dep. of Surgery & 

Translational Medicine, University of Milan-Bicocca, Monza, Italy; 

3 Section of Digestive Diseases, Yale University, New Haven, CT 

Cholangiocarcinoma (CCA) is characterized by a strong 

invasiveness and a dismal prognosis. Early metastasisation to 

regional lymph nodes often precludes surgery, the only potentially 

curative option to treat CCA. Metastatic dissemination 

is facilitated by the presence of an abundant reactive stroma, 

mainly composed by cancer-associated fibroblasts (CAF), and 

by a rich lymphatic vasculature. PDGF-D secreted by CCA cells 

plays a major role in CAF recruitment. Mechanisms of lymphangiogenesis 

in CCA are unknown. Therefore, we investigated 

the possible role of PDGF-D and CAF in this process. Methods/Results. 

Human CCA specimens were immunostained with 

D2-40 (lymphatic endothelial cells, LEC), CD34 (vascular blood 

endothelial cells, VEC), αSMA (CAF) alone or in combination 

with antibodies against VEGF-C or its receptor VEGFR-3 (n=6). 

This analysis showed that in CCA, lymphatic endothelial cells 

(LEC), positive for VEGFR-3, rather than VEC, were closely adjacent 

to VEGF-C expressing CAF. Secretion of VEGF-C, VEGF-D, 

Ang-1, Ang-2 was then evaluated in human primary cultured 

fibroblasts challenged with PDGF-D (n=3). Upon PDGF-D stimulation, 

fibroblasts secreted increased levels of the lymphangiogenic 

growth factor VEGF-C, whereas VEGF-D and Ang-2 

were not expressed. In fibroblasts, PDGF-D-stimulated VEGF-C 

expression was significantly inhibited by blocking PDGFRβ 

(the cognate receptor of PDGF-D) with imatinib mesylate, or 

by inhibiting JNK and ERK/NF-kB signaling with SP600125, 

U0126 and BAY11-7082, respectively. LEC recruitment (tran-


swell migration) and vascular assembly by LEC were measured 

after exposure to conditioned medium (CM) of fibroblasts challenged 

with PDGF-D. CM stimulated LEC recruitment and their 

tubular assembly (lumen diameter, tube elongation and anastomization), 

to an extent comparable to that induced by VEGF-C 

alone. These effects of CM on LEC were inhibited by antagonizing 

PDGFRβ in fibroblasts and by inhibiting VEGFR-3 in LEC 

(n=6). In conclusion, this study provides a working model to 

understand the mechanisms by which the interaction between 

cancer cells and CAF promotes lymphangiogenesis in CCA. 

Our data show that PDGF-D secreted by CCA cells not only 

recruits CAF in the tumor reactive stroma, but also stimulates 

CAF to release VEGF-C acting through the JNK and ERK/NF-kB 

signaling. In turn, VEGF-C promotes tumor lymphangiogenesis 

by stimulating VEGFR-3 expressed on LEC. This interplay may 

be at the basis of the high rate of lymphatic metastasization 

in CCA. Furthermore, the ability to interfere with these mechanisms 

may prevent lymphatic metastatization of CCA. 

Disclosures: 

The following authors have nothing to disclose: Massimiliano Cadamuro, Marta 

Vismara, Simone Brivio, Mario Strazzabosco, Luca Fabris 

Notch1 up-regulation cells expressed both C-myc and OV6 (A, 

white arrow). Representative tumors in liver and lung (B, black 

arrow) and their H&E staining. VCAM-1 was activated in Notch1 

transfected cells by iTRAQ (C, red frame), immunofluoresecence 

(D, white arrow) and western blot analyses (E). VCAM-1 was 

overexpressed (F, black arrow) and TAMs were abnormally 

recruited in the pulmonary metastasis (G, black arrow). 

1951 

Notch-1 Up-regulated Rat Oval Cells Generated Poorly 

Differentiated Liver Cancer and Developed Lung Metastases 

in an Orthotopic Rat Model 

Wen-rui Wu, Xiang-de Shi, Rui Zhang, Lei-bo Xu, Mansheng Zhu, 

Xian-huan Yu, Hong Zeng, Chao Liu; Sun Yat-sen Memorial Hospital, 

Sun Yat-sen University, Guangzhou, China 

Background and aims: Oval cells may give rise to liver cancer. 

Notch signaling has been reported to play crucial role in hepatocellular 

carcinoma. The aim of this study was to investigate 

the biological role of Notch1 in rat oval cell line WB-F344. 

Methods: Notch1 was stably transfected into WB-F344 cells 

by lentivirus. Their proliferation, colony formation, cell cycle 

progression and invasion assays in vitro, as well as tumor 

formation assay in an orthotopic (liver) rat model were performed. 

Results: Gain-of-function analysis showed that Notch1 

up-regulation promoted proliferation, colony formation, G1/S 

cell cycle transition and invasion of WB-F344 cells in vitro. 

Notch1 transfected cells expressed oncogene C-myc and oval 

cell marker OV6 simultaneously. These cells showed a pile up 

appearance in culture and could generate poorly differentiated 

liver cancer orthotopically in isogenic rats (6/6, 100%). 

Interestingly, all tumor bearing rats developed lung metastases 

(6/6, 100%). In addition, iTRAQ analysis indicated that 

vascular cell adhesion molecule 1 (VCAM-1) was activated 

in Notch1 transfected cells in vitro, which was confirmed by 

immunofluorescence and western blot analysis. Moreover, we 

discovered that VCAM-1 was highly expressed in pulmonary 

metastasis tumors and tumor-associated macrophages (TAMs) 

were abnormally recruited in these tumors by immunohistochemistry. 

Conclusions: Notch1 is a crucial regulator of proliferation 

and malignant transformation of oval cells. Notch1 

may facilitate pulmonary metastasis of primary liver cancer via 

VCAM-1 activation and TAMs recruitment. 

Disclosures: 

The following authors have nothing to disclose: Wen-rui Wu, Xiang-de Shi, Rui 

Zhang, Lei-bo Xu, Mansheng Zhu, Xian-huan Yu, Hong Zeng, Chao Liu 

1952 

Deregulated Interplay Between Methionine Adenosyltransferase 

α1, c-Myc and Maf Proteins During Chronic 

Cholestasis Facilitate Development of Cholangiocarcinoma 

Heping Yang 1,2 , Ting Liu 3 , Jiaohong Wang 1,2 , Tony Li 1,2 , Jose M. 

Mato 4 , Shelly C. Lu 1,2 ; 1 Medicine, Cedars-Sinai Medical Center, 

Los Angeles, CA; 2 USC Research Center for Liver Diseases, Los 

Angeles, CA; 3 Department of Gastroenterology, Xiangya Hospital 

Central South University, Changsha, China; 4 Centro de Investigación 

Biomédica en Red de Enfermedades Hepáticas y Digestivas 

(Ciberehd), CIC bioGUNE, Derio, Spain 

Purpose of Study: Cholangiocarcinoma (CCA) develops frequently 

in the setting of chronic inflammation and cholestasis. 

We reported the importance of c-Myc induction in the 

development of cholestatic liver injury and CCA in mice. We 

also showed induction of Maf proteins (MafG and c-Maf) contributed 

significantly to cholestatic liver injury. The aim of our 

current work was to determine whether there is any interplay 

between c-Myc and Maf proteins but in the process of our 

investigation we uncovered interesting reciprocal regulations 

that likely contribute to development of CCA, particularly in 

the setting of chronic cholestasis. Methods: Our study used 

bile duct ligation (BDL) and lithocholic acid (LCA) treatment 

as chronic cholestasis models, diethylnitrosamine followed by 

left and median BDL as a murine CCA model, human CCA 

cell lines KMCH and Huh-28, and human CCA specimens. 

We used immunoprecipitation followed by mass spectrometry 

and recombinant proteins to study protein-protein interactions, 

chromatin immunoprecipitation (ChIP) and sequential ChIP to 

examine co-occupancy of promoter region, overexpression and 

siRNA to vary protein expression. Results: We identified novel 

interacting proteins with c-Myc, specifically methionine adenosyltransferase 

α1 (MATα1, encoded by MAT1A), MafG and 

c-Maf. MAT1A expression fell at the mRNA and protein levels 

in whole liver and bile duct epithelial cells during BDL and LCA 

treatment, and in murine and human CCA samples. The opposite 

occurred with c-Myc, MafG and c-Maf expression. MATα1


interacts mainly with Mnt in normal liver but this switches to 

c-Maf, MafG and c-Myc in cholestatic livers and CCA. Furthermore, 

MATα1 interacts directly with c-Myc, MafG and c-Maf. 

Interestingly promoter regions of MAT1A, c-Maf, MafG and 

c-Myc have E-box sequences that are bound by MATα1 and 

Mnt at baseline that switched to c-Myc, c-Maf and MafG after 

BDL and LCA treatment. MATα1 binds to the E-box as a complex 

with c-Myc and Max, but not by itself or with c-Myc. While 

E-box positively regulates c-Myc, MafG and c-Maf, it negatively 

regulates MAT1A. Furthermore, MATα1 binding to the E-box 

represses E-box and lowers the promoter activity of c-Myc, 

MafG and c-Maf, whereas c-Myc, MafG and c-Maf binding 

enhance E-box-driven promoter activity. This results in reciprocal 

regulation between MATα1 and c-Myc and Maf proteins. 

In CCA cell lines knockdown of MAT1A or overexpression of 

MafG or c-Maf enhanced tumorigenesis. Conclusions: we have 

uncovered a novel interplay between MATα1, c-Myc and Maf 

proteins and their deregulation during chronic cholestasis sets 

the stage for CCA oncogenesis. 

Disclosures: 

The following authors have nothing to disclose: Heping Yang, Ting Liu, Jiaohong 

Wang, Tony Li, Jose M. Mato, Shelly C. Lu 

suppressed by regorafenib in the sorafenib-resistant clones, 

suggesting that sorafenib-resistance might be related to the 

acquisition of resistance against ERK signaling pathway inhibition 

mediated by sorafenib. Promisingly, regorafenib treatment 

suppressed the subcutaneous tumor growth of sorafenib-resistant 

Huh7 and Huh1 clones with significantly prolonged overall 

survival in vivo compared with sorafenib treatment. Conclusions:Regorafenib 

inhibited the growth of sorafenib-resistant 

HCC cells, potentially through suppression of the ERK signaling 

pathway. The efficacy of regorafenib on the sorafenib-resistant 

clones suggests its potential utility in HCC patients with resistance 

to sorafenib. 

Disclosures: 


Mariko Yoshida - Grant/Research Support: Bayer 






The following authors have nothing to disclose: Tomomi Hashiba, Taro Yamashita, 

Kouki Nio, Takehiro Hayashi, Yoshimoto Nomura, Tomoyuki Hayashi, Naoki 

Oishi, Masao Honda 

1953 

Regorafenib inhibits ERK signaling and suppresses the 

growth of sorafenib-resistant cells in human hepatocellular 

carcinoma 

Tomomi Hashiba, Taro Yamashita, Hikari Okada, Kouki Nio, 

Takehiro Hayashi, Yoshimoto Nomura, Mariko Yoshida, Tomoyuki 

Hayashi, Naoki Oishi, Masao Honda, Shuichi Kaneko; Gastroenterology, 

Kanazawa University Graduate School of Medical 

Science, Kanazawa, Japan 

Background:Regorafenib is a multi-kinase inhibitor targeting 

VEGFRs, PDGFRs, and RAF. A recent phase II study suggested 

acceptable tolerability and anti-tumor activity of regorafenib in 

patients with hepatocellular carcinoma (HCC) that progressed 

after initial sorafenib treatment. A confirmatory phase III study 

(NCT01774344) is ongoing. However, it remains unclear 

how regorafenib inhibits the growth signaling pathways of 

sorafenib-resistant HCC. In this study, we evaluated the effects 

of regorafenib and sorafenib on HCC growth and signaling 

pathways in HCC cell lines and primary HCC cells. Methods:Two 

HCC cell lines (Huh1 and Huh7) and three primary 

HCC cells obtained from surgically resected specimens were 

cultured routinely in DMEM supplemented with 10% fetal bovine 

serum and sorafenib tosylate (5-10 μM) for 3 months to generate 

sorafenib-resistant clones. Whole exome sequence analysis 

was performed to detect mutations in the sorafenib-resistant 

clones and their parental cells. Sensitivity against sorafenib 

or regorafenib was evaluated using a cell proliferation assay 

kit. Gene and protein expression was evaluated by quantitative 

RT-PCR and western blotting. A subcutaneous xenotransplantation 

model was used to evaluate the efficacy of orally 

administered sorafenib (30 mgkg -1 day -1 ) or regorafenib (20 

mgkg -1 day -1 ) on sorafenib-resistant clones in vivo. Results:All 

established sorafenib-resistant clones showed higher sensitivity 

to regorafenib compared with sorafenib in vitro, whereas 

parental cells showed similar sensitivity to both sorafenib and 

regorafenib. Sorafenib-resistant clones derived from cell lines 

showed abundant expression of the cancer stem cell marker 

CD44 compared with the parental cells. Although sorafenib-resistant 

clones showed frequent missense/nonsense mutations 

compared with the parental clones, no common mutations were 

detected among all sorafenib-resistant clones. Interestingly, 

ERK phosphorylation was not affected by sorafenib, but was 

1954 

The Dead Box Protein P68 Can Supress The Carcinogenesis 

And Α-Fetoprotein (AFP) Gene Expression via Binding 

The New Silencer Region Of AFP Gene 

Shunsuke Nojiri, Kei Fujiwara, Noboru Shinkai, Etsuko Iio, Takashi 

Joh; Department of Gastroenterology and Metabolism, Nagoya 

City University Graduate School of Medical Sciences, Nagoya, 

Japan 

The activity of the α-fetoprotein (AFP) 

gene decreases rapidly after birth but is reactivated in hepatocellular 

carcinoma (HCC) and also during proliferation of 

hepatocyte. In clinical therapy of hepatocellular carcinoma 

AFP reduction can be the good marker after anticancer therapies. 

We predicted a new silencer lesion that was located 

upstream from previous reported areas. The aim of this study 

is to identify the precise lesion of the silencer and demonstrate 

a new protein which suppresses AFP gene expression via its 

new suppresser lesion. A basic vector was constructed 

by inserting the 4.9-kb AFP5’-flanking sequence into 

PGL2 basic vector. The vectors deleted several sections were 

also constructed. The Luciferase activities were expressed in 

relation to the activity of the cells that were transfected with 

the basic vector and we found the 179 base pair new silencer 

lesion. After immunoprecipitation by biotinized PCR products 

including the silencer arrangement and the nuclear extracts, the 

protein was identified from using an LC-MS/MS assay. Chromatin 

immunoprecipitation (ChIP) was performed to confirm the 

protein binding to the new silencer. The candidate protein was 

suppressed or overexpressed using siRNA duplexes or overexpression 

vector in HepG2 cells and AFP expression measured 

by Luciferase activities and real time PCR and cell proliferation 

were measured by cell proliferation assay. The results 

of the LC-MS/MS assay demonstrated the presence of DEAD 

box protein p68 (p68). P68 protein could binding to the new 

silencer session was confirmed by ChIP. After using RNAi to 

p68 the Luciferase activities increased 3.5±1.1 times (p


same results. A new 179 base pair silencer position 

was identified which was located at 1153 bases upstream 

from which was reported previously. The p68 protein could 

suppress the AFP gene expression via binding to this lesion 

directly and suppress the cell proliferation. In hepatocellular 

carcinoma cells p68 may suppress not only AFP production 

but also cell proliferation. In the clinical therapies reduction 

of AFP can become the good prediction of carcinogenesis or 

reduction the mass of carcinoma. This report revealed the one 

aspect of the mechanism of these relationships. 

Disclosures: 

The following authors have nothing to disclose: Shunsuke Nojiri, Kei Fujiwara, 

Noboru Shinkai, Etsuko Iio, Takashi Joh 

1955 

WITHDRAWN 

1956 

Macrophage colony-stimulating factor receptor antagonist 

inhibits progression of hepatocellular carcinoma in 

vivo. 

Hiroshi Kono, Yoshihiro Akazawa, Shinji Furuya, Hideki Fujii; First 

Department of Surgry, University of Yamanashi, Chuo, Japan 

Aim; We previously reported that macrophage colony-stimulating 

factor (M-CSF) is involved in hepatocarcinogenesis by 

inducing an angiogenic factor via the hepatic Mφs. Therefore, 

M-CSF could be targets of therapy in hepatocellular carcinoma 

(HCC). The spurpose of this study was to investigate effects of 

M-CSF receptor antagonist on HCC. Materials and Method; 

1; isolated mouse hepatic macrophages (Mφs) or monocytes 

(Mos) were cultured with media added the different dose of 

M-CSF. Production of vascular endothelial growth factor (VEGF) 

was assessed. Furthermore, isolated vascular endothelial cells 

(VEC) were co-cultured with or without hepatic Mφs in presense 

with M-CSF (100ng/ml) for 5 days and cell proliferations were 

assessed in vitro. 2; C57/BL6 mice were treated with diethyl 

nitrosamine (DEN) intraperitoneally. For treatment group, 

M-CSF receptor antagonist (GW2580) was treated every day. 

Incidence of tumors was assessed 38 weeks after treatment. 

Furthermore, angiogenesis and distribution of CD163-positive 

M2 Mφs were assessed. 3; Mouse HCC cells (MH134) were 

implanted to same strain C3H mice by subcutaneous injection 

(1 ×10 5 /animal). Tumor progression was assessed after 

3 weeks. 4; In the nude mouse, human HCC cells (HuH7) 

were implanted by intra-splenic injection (1 × 10 6 /animal). 

Tumor progression in the spleen was assessed after 3 weeks. 

5; MH134 or HuH7 (1 × 10 4 /well) was cultured with media 

containing M-CSF (100ng/ml) in the presence or absence of 

GW2580 (1ng/ml) for 7 days in vitro, and cell proliferation 

was assessed. Result; 1; Production of VEGF increased both in 

hepatic Mφs and MOs incubated with M-CSF in dose dependent 

and time dependent manner. Furthermore, the production 

was significantly greater in hepatic Mφs compared with that in 

Mos. Importantly, proliferation of VEC was greatest in cells cultured 

with hepatic Mφs in media with M-CSF. 2; Hepatic tumors 

diagnosed as hepatocellular adenoma or HCC were observed 

in animals treated with DEN. In contrast, tumor incidence was 

significantly reduced in DEN-treated animals with GW2580. 

Enhanced angiogenesis M2Mf population observed in the 

DEN-treated animals was significantly blunted by treatment of 

GW2580. 3 and 4; Growth of implanted both of HCC was 

significantly inhibited by GW2580 in vivo. 5; Proliferations 

both of HCC were not significant difference between in cells 

cultured with M-CSF and cells cultured without M-CSF. Furthermore, 

GW2580 did not inhibit proliferation of these cells in 

vitro. Conclusions; M-CSF receptor antagonist markedly inhibited 

tumor initiation and progression. Thus, M-CSF and/or its 

receptor could be a new therapeutic target for HCC. . 

Disclosures: 

The following authors have nothing to disclose: Hiroshi Kono, Yoshihiro Akazawa, 

Shinji Furuya, Hideki Fujii 

1957 

Promotion of Hepatocarcinogenesis by the Endocannabinoid 

Anandamide and its Receptor CB1 

Ki Tae Suk 1 , Ingmar Mederacke 1 , Geum Youn Gwak 2 , Robert F. 

Schwabe 1 ; 1 Columbia University, New York, NY; 2 Sungkyunkwan 

University, Seoul, Korea (the Republic of) 

BACKGROUND: The endocannabinoid system (ECS) exerts key 

roles in the development of liver fibrosis and fatty liver, two 

diseases that promote hepatocarcinogenesis. Although cannabinoids 

exert potent anti-tumor effects in vitro, the role of the 

ECS in carcinogenesis in vivo remains elusive. AIM: To understand 

the contribution of the ECS to hepatocarcinogenesis. 

METHODS: Expression endocannabinoids (EC), EC-degrading 

enzymes and EC receptors, was determined by LC-MS/MS, 

qPCR and immunohistochemistry in mouse and patient samples. 

The contribution of ECS to diethylnitrosamine (DEN)-induced 

hepatocellular carcinoma (HCC) was determined in 

mice deficient in fatty acid amide hydrolase (FAAH), the main 

anandamide (AEA)-degrading enzyme, and in mice deficient 

for cannabinoid receptor 1 (CB1), cannabinoid receptor 2 

(CB2), or transient receptor potential cation channel subfamily 

V member 1 (TRPV1). RESULTS: Murine and human HCCs displayed 

activation of the ECS with elevated expression of CB1 

and CB2 mRNA and protein, unaltered TRPV1 expression and 

only moderate alterations of EC levels. Surprisingly, FAAH-deficient 

mice, which have about 10-fold increased hepatic levels 

of CB1 agonist AEA, displayed a 129% (p


1958 

Chronic alcohol accelerates early hepatobiliary cancer 

by increasing inflammation, stemness, EMT and miR- 

122 mediated HIF-1α activation 

Aditya Ambade, Abhishek Satishchandran, Gyongyi Szabo; Medicine, 

UMass Medical School, Worcester, MA 

Background / Aims: Chronic alcohol use is a major risk factor 

in the development of hepatocellular carcinoma [HCC]. 

Inflammation contributes to alcoholic liver disease, particularly 

in alcoholic hepatitis, and promotes HCC development. Other 

molecular events leading to HCC include proliferation of tumor 

stem cells, epithelial-mesenchymal transition (EMT) and altered 

microRNA expression in the liver. We hypothesized that the 

pro-inflammatory microenvironment in alcoholic hepatitis accelerates 

liver tumor development after pre-exposure to a chemical 

carcinogen. Methods: Four week old male C57BL/6 mice 

were injected intraperitoneally with N, N diethyl nitrosamine 

(DEN), 75 mg/kg/week for 3 consecutive weeks followed by 

100 mg/kg/week for 3 additional weeks. At age 8 weeks, 

mice were divided into alcohol (4% Lieber-DeCarli alcohol 

diet) or calorie matched control diet (pair-fed group). After 6 

weeks of alcohol feeding mice were sacrificed; blood, liver 

tissues were collected for analysis of cytokines, tumor markers 

and miR-122. Cyclin D1, p53, vimentin and sonic hedgehog 

proteins were evaluated by western blots. HIF-1a activity was 

assayed by EMSA. Results: Alcohol-fed DEN-injected mice 

had greater liver damage (ALT) and significantly increased 

liver-to-body weight ratio compared to pair-fed DEN-injected 

mice. Alcohol feeding resulted in steatohepatitis indicated by 

increased pro-inflammatory cytokines TNFa, IL-6, MCP-1, IL-17 

and fibrotic genes a-SMA, procollagen1a1 and TGFb, that 

were significantly higher in alcohol plus DEN mice compared 

to other groups. MRI and liver histology of alcohol plus DEN 

mice revealed hepatobiliary cysts, early hepatic neoplasia and 

increased AFP. No tumors were found in other groups. Proliferation 

makers (PCNA, cyclin D1, p53), EMT markers (vimentin, 

sonic hedgehog) and cancer stem cell markers (CD133 

and nanog) were significantly upregulated in livers of alcohol-fed 

DEN-injected mice compared to pair-fed DEN-injected 

and alcohol-fed saline-injected mice. Immunostaining for AFP, 

PCNA, cytokeratin7 and 19 showed increased number of 

stained cells in alcohol-fed DEN-injected mice compared to 

all other groups indicating proliferation of hepatobiliary progenitors. 

In livers with tumors from alcohol plus DEN mice, we 

found significant reduction of miR-122 with upregulation of 

miR-122 target gene, HIF-1α. Increased HIF-1a DNA binding 

activity and upregulation of its target, VEGFR1, was found in 

livers of alcohol plus DEN mice compared to all other groups. 

Conclusion: Our findings establish a new model of early HCC 

and demonstrate that alcoholic hepatitis accelerates hepatobiliary 

tumor development with molecular features of HCC. 

Disclosures: 

The following authors have nothing to disclose: Aditya Ambade, Abhishek Satishchandran, 

Gyongyi Szabo 

1959 

Variations of the host genome and interaction of hepatitis 

B viral X protein associated with hepatocarcinogenesis 

Hiroko Nagata 1 , Yasuhiro Itsui 1,2 , Fukiko Kawai-Kitahata 1 , Shun 

Kaneko 1 , Miyako Murakawa 1,2 , Sayuri Nitta 1 , Mina Nakagawa 1 , 

Seishin Azuma 1 , Sei Kakinuma 1 , Yasuhiro Asahina 1 ; 1 Department 

of Gastroenterology and Hepatology, Tokyo Medical and Dental 

University, Tokyo, Japan; 2 Department of Medical Education 

Research and Development, Tokyo Medical and Dental University, 

Tokyo, Japan 

Background and Aims: Association between variation of the 

host genome and viral proteins responsible for hepatocarcinogenesis 

is unclear. To clarify this, we comprehensively analyzed 

variations of the host genome in patients with HCC using 

a next-generation sequencer, and investigated the association 

between identified host genome variation and HBx protein. 

Methods: [1] We performed deep sequenceing in 104 pairs of 

HCC and non-tumor samples targeting 54 cancer-related genes 

containing 2820 mutational hotspots. [2] To assess the effect of 

TERT promoter variations (C228T, C250T), which were identified 

as most frequent mutations in our samples, on its transcriptional 

activity, reporter assays using TERT promoter-luciferase 

plasmids with or without these mutations were performed. 

[3] To assess the association between TERT promoter activity 

and HBx protein, reporter assays were performed 48 hours 

after co-transfection of plasmid expressing HBx protein and 

TERT promoter-luciferase plasmids in HepG2 cells and 293T 

cells. [4] Moreover, effects of transcriptional factor Sp1 and 

c-myc on TERT promoter activity were analyzed by additional 

co-transfection using plasmid expressing these transcriptional 

factors. Results: [1] We detected somatic mutations in 9 genes 

out of 93 samples: TERT, TP53, CTNNB1, PTEN, CDKN2A, 

HRAS, PIK3CA, STK11, GNAS and NFE2L2. Among them, 

TERT promoter mutations were most frequently found (65% of 

the patients) in overall. However, these mutations were significantly 

less frequent in HBV-related HCC, while integration of 

HBx gene into TERT region was frequently found, which was 

mutually exclusive to TERT promoter mutations. [2] The TERT 

mutations (C228T, C250T) significantly upregulated the promoter 

activity in comparison with wild-type prompter in HepG2 

and 293T cell (181±2.2%, 255±5.2%). [3] Expression of HBx 

protein resulted in further activation of TERT promoter activity 

in both of TERT-mutant and wild type (714±3.1%, 628±2.1%), 

and effect of HBx protein on TERT promoter activation was 

more remarkable in the TERT-wild promoter than the TERT-mutant 

one (560%, 435%). [4] The promoter activity was elevated 

by Sp1 or c-myc, which are TERT promoter-binding factors. 

Conclusions: TERT promoter mutations were most frequently 

detected in non-HBV related HCC, but rare in HBV-related 

HCC, in which integration of HBx into TERT region were frequently 

and exclusively found. Upregulation of TERT promoter 

activity by either TERT promoter mutations or interaction of HBx 

protein with TERT promoter via Sp1 and c-myc may associate 

with hepatocarcinogenesis or tumor progression. 

Disclosures: 


MSD Co., Ltd. 

The following authors have nothing to disclose: Hiroko Nagata, Yasuhiro Itsui, 

Fukiko Kawai-Kitahata, Shun Kaneko, Miyako Murakawa, Sayuri Nitta, Mina 

Nakagawa, Seishin Azuma, Yasuhiro Asahina


1960 

Survival of hepatocellular carcinoma patients according 

to recommended treatment by the Barcelona Clinic Liver 

Cancer or Hong Kong Liver Cancer staging system in 

hepatitis B endemic area 

Ju-Il Yang, Dong Hyun Sinn, jung hee kim, Geum-Youn Gwak, 

Yong-Han Paik, Moon Seok Choi, Joon Hyeok Lee, Kwang Cheol 

Koh, Seung Woon Paik, Byung Chul Yoo; Gastroenterology, Samsung 

Medical Center, Seoul, Korea (the Republic of) 

Background: Cancer staging system aims to help to predict 

patient prognosis as well as select treatment modality. Several 

staging system has been proposed for hepatocellular carcinoma 

(HCC), and of those, only Barcelona Clinic Liver Cancer 

(BCLC) and Hong Kong Liver Cancer (HKLC) staging system 

has recommended treatment modality. We analyzed long-term 

outcome of patients according to the recommended treatment 

modality in both staging system. Methods: A total of 3,515 treatment-naïve, 

newly diagnosed HCC patients [age: 62.8 ± 10.5, 

male: 2,792 (79.4%), hepatitis B virus: 2,709 (77.1%)] in a 

single center were analyzed. Results: The median survival was 

5.4 years, with 5-years cumulative survival rate of 50%. BCLC 

and HKLC staging well-stratified patients prognosis (5-years 

survival rate: 79.1%, 62.9%, 40.3%, 21.3%, and 27.0% for 

BCLC 0, A, B, C and D, respectively; 72.3%, 54.9%, 50.6%, 

21.3%, 10.2%, 16.7%, 7.2%, 47.1% and 11.3% for stage 

I, IIa, IIb, IIIa, IIIb, IVa, IVb, Va and Vb, respectively). C-index 

of BCLC and HKLC staging system was 0.708 and 0.732. 

Overall, 49.5% patients received BCLC-recommended treatment, 

and patients survival was better when patient received 

recommended treatment in stage O and A, but survival was 

worse in stage B, C and D. For HKLC staging system, 55.6% of 

patients received HKLC-recommended treatment, and survival 

was better when patient received recommended treatment in 

stage I, IIa, IIb, IIIa, Va but not in stage IIIb, IVa, IVb, and Vb. 

Conclusion: Both BCLC and HKLC staging system could stratify 

patient prognosis, however, could not direct therapy for large 

proportion of patients, and in some stage, recommended therapy 

was associated with worse prognosis. 

Disclosures: 

The following authors have nothing to disclose: Ju-Il Yang, Dong Hyun Sinn, jung 

hee kim, Geum-Youn Gwak, Yong-Han Paik, Moon Seok Choi, Joon Hyeok Lee, 

Kwang Cheol Koh, Seung Woon Paik, Byung Chul Yoo 

1961 

Inactivation of Aldose Reductase Prevents NAFLD, 

NASH, and Hepatocarcinogenesis in Transaldolase Deficiency 

Zachary Oaks 1 , Robert Hanczko 1 , Miguel Beckford 1 , Sookja K. 

Chung 3 , Steve Landas 1 , John Asara 2 , Andras Perl 1 ; 1 Medicine, 

SUNY Upstate, Syracuse, NY; 2 Medicine, Beth Israel Deaconess 

Medical Center, Boston, MA; 3 Anatomy, The University of Hong 

Kong, Hong Kong, China 

Background and Aims: Transaldolase (TAL) is a rate limiting 

enzyme of the pentose phosphate pathway. TAL deficiency 

elicits NAFLD, NASH, and hepatocellular carcinoma (HCC) 

in humans and mice. The pathogenesis is driven by oxidative 

stress and characterized by the accumulation of sugar phosphates, 

depletion of NADPH and glutathione (J. Clin. Invest. 

2009;119:1546-57). TAL-deficient mice exhibit the overexpression 

of the NADPH dependent enzyme aldose reductase 

(AR). Here, we investigated if the role of AR in the depletion of 

NADPH and the resultant pathologies. Methods: Metabolites 

were extracted from TAL, AR, and TAL/AR double-knockout 

livers that were sacrificed along with C57BL/6 controls, using 

5 age-matched mice of each strain. 258 metabolites were 

detected by LC-MS/MS with selective reaction monitoring. 

Relative metabolite concentrations and pathway fluxes were 

evaluated using Metaboanalyst. In addition, the development 

of NAFLD, NASH, and HCC were monitored up to 78 weeks 

of age. Results: 0/21 TAL/AR double-knockouts, 45/97 TAL 

knockouts, and 3/106 controls developed HCC by 78 weeks 

of age. Deletion of AR in TAL deficiency also rescued fatty liver; 

no hepatosteatosis was detected in mice lacking both enzymes. 

AR knockout livers had no significant pathology relative to 

control animals. The liver of double-knockouts were similar 

to wild-type from controls (p=1.0) and they had significantly 

reduced HCC compared to TAL knockouts (p


This study suggests that TGFβ1 is involved in CCA tumor progression 

and may mediate this process through miR-34a downstream 

signaling pathways, raising the possibility that TGFβ1 

signaling may offer potential therapeutic targets to inhibit CCA 

development and growth. 

Disclosures: 

The following authors have nothing to disclose: Chiung-Kuei Huang, Arihiro 

Aihara, Yoshifumi Iwagami, Tunan Yu, Rolf I. Carlson, Hironori Koga, Miran 

Kim, Jack R. Wands 

1963 

Hepatic Ruvbl1 is key regulator of glucose homeostasis 

and promotes hepatocellular carcinoma progression in 

vivo. 

Tommaso Mello 1,2 , Francesca Zanieri 1 , Elisabetta Ceni 1 , Mirko 

Tarocchi 1 , Giada Marroncini 1 , Simone Polvani 1 , Sara Tempesti 1 , 

Oxana Bereshchenko 3 , Stefano Milani 1,2 , Andrea Galli 1,2 ; 1 University 

of Florence, Florence, Italy; 2 Careggi University Hospital, 

Florence, Italy; 3 University of Perugia, Perugia, Italy 

The AAA+ ATPase Ruvbl1 is overexpressed in several human 

cancers, including hepatocellular carcinoma (HCC), in which 

high Ruvbl1 expression correlates with a poor prognosis. A 

growing body of data from in vitro models supports the concept 

that deregulation of Ruvbl1 expression occurs in cancer to promote 

its growth and progression, making this protein an attractive 

target for anti-cancer therapies. However, whether Ruvbl1 

actively drives HCC onset and progression remains speculative. 

To challenge this question we realized an hepatocyte-conditional 

Ruvbl1 +/- mouse and evaluated the number and size 

of tumor foci induced by DEN. Ruvbl1 +/- mice were obtained 

by crossing Ruvbl1-floxed with Albumin-Cre deleter mice. The 

male offspring were subjected to a single i.p. injection of DEN 

(5mg/kg) to induce liver cancer, and were monitored up to one 

year after cancer induction. Contrary to our expectations, we 

found that despite an initial delay in the appearance of liver 

cancer 6 month after DEN injection, conditional Ruvbl1 +/- mice 

eventually developed larger tumors that control floxed mice 

9 and 12 months after tumor induction. Ruvbl1 +/- mice (not 

DEN-injected) showed impaired basal hepatocyte turnover and 

strikingly lower regeneration after 70% hepatectomy, confirming 

that Ruvbl1 support cell growth in vivo. We observed that 

in the hemizygous mice Ruvbl1 expression was reduced by 

50% in normal hepatocytes but not within the tumors, which 

indeed overexpressed Ruvbl1 regardless of the mouse genetic 

background, suggesting that Ruvbl1 overexpression is a central 

event in HCC onset. Hepatic-Ruvbl1 +/- mice developed mild 

hyperglycemia, impaired glucose tolerance, insulin resistance, 

had reduced liver glycogen content and increased somatic 

growth. Moreover, mRNA expression of several mTOR targets 

(SREBP-1c, HIF1α, PGC-1α, PPARγ) were reduced in Ruvbl1 +/- 

mice. Ruvbl1 silencing in non-transformed hepatocytes (AML- 

12) resulted in reduced mTOR expression and impaired 

PI3K-Akt-mTOR signaling in response to insulin, as well reduced 

cell growth. By 2D-DIGE proteomics we identified key enzymes 

of glycolysis as down-regulated proteins in Ruvbl1-silenced 

hepatoma cells. Taken together these results highlight a novel 

role of Ruvbl1 in the maintenance of hepatic glucose homeostasis 

and insulin-sensitivity through the Akt-mTOR pathway, and 

strongly suggest that deregulation of this pathway by Ruvbl1 

overexpression is a key event driving HCC progression. 

Disclosures: 

Stefano Milani - Grant/Research Support: Gilead Sciences, Merck Sharp and 

Dohme, Abbvie 

The following authors have nothing to disclose: Tommaso Mello, Francesca Zanieri, 

Elisabetta Ceni, Mirko Tarocchi, Giada Marroncini, Simone Polvani, Sara 

Tempesti, Oxana Bereshchenko, Andrea Galli 

1964 

MicroRNA-207 controls Prp19 expression in hepatocellular 

carcinoma 

Ji-Min Zhu, Xi-Zhong Shen; Department of Gastroenterology, 

Zhongshan Hospital of Fudan University, Shanghai, China 

INTRODUCTION: microRNAs (miRNAs) are small non-coding 

RNA molecules of 21-24 nt that regulate the expression of 

numerous target genes by transcriptional inhibition or translational 

repression. Multiple lines of evidence suggest that miR- 

NAs play important roles in tumor (such as, hepatocellular 

carcinoma (HCC)) development, progression, invasion, metastasis 

and prognosis. Our previous unpublished work suggested 

that microRNA-207 (miR-207) upregulates in HCC tissues than 

in normal liver tissues. Unfortunately, our understanding of the 

molecular mechanisms that governs its role in development, 

progression, invasion, metastasis and prognosis remains fragmentary. 

AIMS & METHODS: In this study, a genome-wide 

miRNA microarray was used to identify differentially expressed 

miRNAs in HCCs patients. Next, miR-207 expression in normal 

liver tissues, HCC tissues and adjacent non-tumor tissues was 

validated, and the predictive values of miR-207 for the prognosis 

of HCC patients were explored using χ2 test, Student’s 

t test, paired t test, and Mann-Whitney test. The biological 

roles of miR-207 and its underlying roles in HCC were also 

investigated in the Huh7, MHCC-97H, SMMC-7721, L02 and 

Hep3B cell lines. RESULTS: Forty-two miRNAs were differentially 

expressed in HCCs. Several of these miRNAs were previously 

found deregulated in other cancers. Additionally, the 

miR-207 was found upregulated in ∼80% of HCCs and in all 

HCC-derived cell lines. Overexpressed intratumoral miR-207 

was associated with poor survival rate (P < 0.001), and was 

an independent prognostic factor for overall survival rate (P < 

0.001) in the patients with HCC. Overexpression of miR-207 in 

L02 cell line remarkably enhanced cell proliferation, migration, 

and invasion; while inhibition of miR-207 in Hep3B cell line 

caused the opposite effects. Further study found that miR-207 

suppressed the expression of pre-Mrna processing factor 19 

(Prp19) by directly binding to its 3’-untranslated region, and 

sensitized Hep3B cell to CDDP-induced apoptosis. Moreover, 

miR-207 expression levels correlated positively with Prp19 in 

human HCC tissues. Western blot showed that overexpression 

of miR-207 in L02 cell upregulates the expression of Prp19, 

while inhibition of miR-207 in Hep3B cell downregulates the 

expression of Prp19, suggesting that Prp19 might play as a 

oncogene in HCC. CONCLUSION: Taken together, our findings 

suggest that miR-207 could play a role in the development of 

HCC, at least in part, by modulating Prp19. These findings 

establish a basis toward the development of therapeutic strategies 

aimed at blocking miR-207 in HCC. 

Disclosures: 

The following authors have nothing to disclose: Ji-Min Zhu, Xi-Zhong Shen


1965 

Somatic Mutational Analysis by Next Generation RNA 

Sequencing Reveals Frequent Mutation of Chromatin 

and Chromosome Remodeling Genes in Gallbladder 

Cancer 

Loretta K. Allotey 1 , Roongruedee Chaiteerakij 1,2 , Gavin R. Oliver 

3 , Vivekananda Sarangi 3 , Renumathy Dhanasekaran 1 , Catherine 

D. Moser 1 , Nasra H. Giama 1 , Daniel R. O’Brien 3 , Raymond 

M. Moore 3 , Mia D. Champion 4 , Eric W. Klee 3 , Mitesh J. Borad 5 , 

Lewis R. Roberts 1 ; 1 Division of Gastroenterology and Hepatology, 

Mayo Clinic College of Medicine, and Mayo Clinic Cancer Center, 

Rochester, MN; 2 Department of Medicine, Faculty of Medicine, 

Chulalongkorn University and King Chulalongkorn Memorial Hospital, 

Thai Red Cross Society, Bangkok, Thailand; 3 Department of 

Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN; 

4 Department of Biomedical Statistics and Informatics, Mayo Clinic, 

Scottsdale, AZ; 5 Division of Hematology and Medical Oncology, 

Mayo Clinic, Scottsdale, AZ 

Background/Aim: Gallbladder cancer (GBC) is uncommon, 

with an incidence of 2 per 100,000/year in the US. Due 

to its asymptomatic nature, most patients present at a late 

stage. Current standard chemotherapy and radiation therapy 

only achieve a 5-year survival rate of 10% in patients with 

advanced cancer. Thus improved understanding of the molecular 

pathogenesis of GBC is urgently needed for rational design 

of effective therapies. We therefore aimed to investigate the 

mutational landscape of human gallbladder cancer. Methods: 

12 GBCs were RNA sequenced on the Illumina Hi-Seq 2000 

platform. Single Nucleotide Variants (SNVs) were called from 

the genome-aligned reads using Unified Genotyper. Stringent 

filtering criteria were used to identify variants of high 

confidence (Inclusion criteria: Depth of Coverage>10, Quality 

Score>0.05, Minor Allele Frequency1 

patient). To identify variants in clinically relevant genes, variants 

meeting the above criteria were selected and compared to 

a generated list of 1,434 genes reported in COSMIC, Cancer 

Panels, or in the literature to have relevance to cancer and epigenetics 

processes. Survival in months was determined for each 

patient and select genes were investigated for their association 

with survival by Kaplan Meier analysis. The significance of the 

differences in survival between groups was analyzed using the 

Log-Rank test. Results: 69 SNVs were identified in 28 clinically 

relevant genes. Fifteen of the 28 genes (53%) have known 

effects on epigenetic processes, including TP53 (mutated in 

50% of tumors), SETD1A (33%), MLL2 (33%), SRCAP (25%), 

MSH6 (16%), MSL1 (16%), ATXN7 (16%), SUV420H1 (16%), 

LSG1 (16%), RSF1 (16%), MCM7 (16%), DNAJC2 (16%), 

AHCTF1 (16%), BRD4 (16%), and NUP214 (16%). DAVID 

Functional annotation and WebGestalt Gene Ontology Analysis 

of the 28 genes revealed that 11 genes (ATXN7, SRCAP, 

DNAJC2, MSL1, SUV420H1, BRD4, MLL2, SETD1A, TP53, 

and RSF1) are involved in chromatin and chromosome remodeling. 

Of the 11 genes, mutations in the SRCAP gene were 

found to have a marginally significant effect on survival. The 

three patients with mutation in the SRCAP gene had a survival 

of 4 months while patients without mutations in SRCAP had 

a survival of 9 months (P = 0.06). Conclusions: Chromatin 

and chromosome remodeling genes are frequently mutated in 

GBC and provide a starting point for further functional studies. 

Patients with SRCAP mutations show a trend towards poorer 

survival than patients with wild-type SRCAP. Validation of these 

findings in a larger cohort is underway. 

Disclosures: 




The following authors have nothing to disclose: Loretta K. Allotey, Roongruedee 

Chaiteerakij, Gavin R. Oliver, Vivekananda Sarangi, Renumathy Dhanasekaran, 

Catherine D. Moser, Nasra H. Giama, Daniel R. O’Brien, Raymond M. Moore, 

Mia D. Champion, Eric W. Klee, Mitesh J. Borad 

1966 

Differential Antitumor Effects of FGFR Inhibitors on a 

Novel Cholangiocarcinoma Patient-Derived Xenograft 

Mouse Model Expressing an FGFR2-CCDC6 Fusion Protein 

Yu Wang 1,2 , Hassan M. Shaleh 2 , Xiwei Ding 2,3 , Shaoqing 

Wang 2,4 , Roongruedee Chaiteerakij 2,5 , Kais Zakharia 2 , Loretta 

K. Allotey 2 , Essa A. Mohamed 2 , Catherine D. Moser 2 , Steven 

Alberts 6 , Joseph M. Gozgit 7 , Mitesh J. Borad 8 , Lewis R. Roberts 2 ; 

1 Department of Hepatobiliary Surgery, Nanfang Hospital, Southern 

Medical University, Guangzhou, China; 2 Division of Gastroenterology 

and Hepatology, Mayo Clinic College of Medicine, and 

Mayo Clinic Cancer Center, Rochester, MN; 3 Department of Gastroenterology, 

Drum Tower Hospital, Affiliated to Medical School 

of Nanjing University, Nanjing, China; 4 Department of Pathology, 

Qiqihar Medical University, Qiqihar, China; 5 Department of Medicine, 

Faculty of Medicine, Chulalongkorn University and King 

Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, 

Thailand; 6 Department of Oncology, Mayo Clinic, Rochester, 

MN; 7 ARIAD Pharmaceuticals, Inc., Cambridge, MA; 8 Division of 

Hematology and Medical Oncology, Mayo Clinic, Scottsdale, AZ 

Introduction: Cholangiocarcinoma (CCA) is a highly lethal 

cancer with limited therapeutic options and a poor prognosis. 

Recent genomic analysis has revealed the presence of 

FGFR2 fusion proteins in up to 13% of intrahepatic CCAs. 

The FGFR fusion proteins appear to play a key role in cancer 

progression. Tumors harboring FGFR fusions have demonstrated 

enhanced sensitivity to FGFR inhibitors, suggesting that 

CCA patients with FGFR2 fusions may benefit from targeted 

FGFR2 kinase inhibition. The effects of FGFR kinase inhibitors 

ponatinib, dovitinib and BGJ398 have not been evaluated in 

CCA bearing FGFR fusions. We aimed to assess the relative 

sensitivity of a novel in vivo CCA PDX mouse model bearing 

an FGFR2-CCDC6 fusion protein to different FGFR inhibitors. 

Methods: LIV31 is a PDX model derived in NSG SCID mice 

from a metastatic lung nodule resected from a patient with 

stage IV intrahepatic CCA. Subsequent passage of the PDX 

tumor into nude mice was successful. Detailed assay confirmed 

that LIV31 harbors a transcript encoding an FGFR2-CCDC6 

fusion protein. LIV31 cells were implanted subcutaneously into 

the flanks of nude mice. When tumor volumes reached 150- 

250 mm3 the mice were randomized into four groups: vehicle, 

ponatinib 25 mg/kg, dovitinib 30 mg/kg, or BGJ398 15 

mg/kg. The drugs were administered daily by oral gavage at 

doses previously shown to be tolerated by mice. Tumor volumes 

and mouse weights were measured once a week. Tumor 

growth curves were compared. After 63 days treatment, the 

animals were euthanized and xenografts were examined by 

histology, Ki-67 IHC, Tunel staining and Western blotting for 

pFGFR2 and the FGFR pathway downstream mediators pFRS2, 

pAKT, and pERK. Results: All three FGFR inhibitors significantly 

inhibited the growth of the FGFR2-CCDC6 fusion mouse PDX 

tumors when compared with vehicle (P


The Ki-67 and Tunel staining results show that these inhibitors 

can also inhibit proliferation and induce apoptosis of the CCA 

xenograft cells. Conclusions: These three TKIs all significantly 

inhibited the growth of patient-derived CCA xenografts bearing 

an oncogenic FGFR2-CCDC6 fusion gene. Treatment efficacy 

from most to least effective was BGJ398 > dovitinib > ponatinib. 

Additional studies exploring the potential mechanisms 

underlying differences in response to particular FGFR inhibitors 

and the effects of combinations of other therapeutic agents with 

FGFR inhibitors are underway. 

Disclosures: 

Joseph M. Gozgit - Employment: ARIAD Pharmaceuticals 




The following authors have nothing to disclose: Yu Wang, Hassan M. Shaleh, 

Xiwei Ding, Shaoqing Wang, Roongruedee Chaiteerakij, Kais Zakharia, Loretta 

K. Allotey, Essa A. Mohamed, Catherine D. Moser, Steven Alberts, Mitesh J. 

Borad 

1967 

Glypican 3 contributes to HCC invasiveness and tumorigenesis 

by up-regulating Angioprotein 2 and initiating 

epithelial mesenchymal transition 

Thu Le Trinh, William M. Puszyk, Olorunseun Ogunwobi, Quanfeng 

Wu, Chen Liu; Department of Pathology, University of Florida, 

Gainesville, FL 

Background and Rationale: Glypican 3 (GPC3) has been 

shown to be up-regulated in HCC and is known to be associated 

with HCC advanced stages as well as cancer invasiveness 

and recurrence. The aim of this study is to define the 

mechanisms of GPC3 in HCC carcinogenesis and metastasis. 

Methods: Representations of murine and human HCCs were 

created using GPC3 knock-in in 1MEA (1MEA hGPC3 ), a murine 

HCC cell line which doesn’t have GPC3 expression; and GPC3 

knock-down in Huh7 (Huh7 GPC3 shRNA ), a human HCC cell line 

which highly expresses GPC3. Cell proliferation and tumor 

development were access in vitro and in vivo using the 1MEA/ 

Balb/c and Huh7/NSG mice models. The impact of GPC3 

knock down on different cancer pathways was approached by 

qPCR array analysis on the Huh7 control shRNA and GPC3-knockdown 

Huh7 GPC3 shRNA cell lines using Cancer Pathway Finder 

array (Qiagen), which consists of 84 different genes that represent 

9 different biological pathways involved in carcinogenesis. 

The effect of GPC3 repression on cell invasion ability 

was also examined by wound-healing assay. Results: We 

found that GPC3 didn’t induce a different proliferation rate 

in 1MEA hGPC3 cell line, but it promoted tumor development 

significantly when 1MEA hGPC3 was implanted into Balb/c mice 

compared to 1MEA control. Knock-down of GPC3 resulted in 

cell growth inhibition and cell cycle arrest at S phase. Similarly, 

tumor proliferation was greatly retarded in Huh7 GPC3 

shRNA 

xenograft mouse model in comparison to Huh7control shRNA 

tumor. In the 84 cancer pathway focused genes, 29 genes 

showed at least a 2-fold difference in gene expression between 

GPC3-knockdown cells compared to non-silenced cells. These 

included genes that maintain telomeres (TINKS, TINKS2, TINF2, 

TERF2IP), anti-apoptotic genes (XIAP, NOL3, FASLG), genes 

involved in cell cycle regulation (CCND2, CCND3, E2F4), 

markers of epithelial to mesenchymal transition (EMT) (CDH2, 

DSP, OCLN, SNAI3), and genes that promote angiogenesis 

(ANGPT2). Among these, ANGPT2 gene expression was 

down-regulated up to 150 fold in Huh7 shRNA GPC3 compared to 

Huh7 control shRNA . GPC3 roles in EMT were confirmed by migration 

assay and showed significant inhibition of Huh7GPC3 shRNA 

migration. Likewise, EMT markers and inducers were upregulated 

in 1MEA hGPC3 cell line in comparison to 1MEA control. 

Conclusions: These results indicated that GPC3 played a role 

in HCC tumorigenesis via promoting EMT, angiogenesis and 

cell migration, and that GPC3 could serve as a key connection 

between angiogenesis and metastasis in HCC. 

Disclosures: 

The following authors have nothing to disclose: Thu Le Trinh, William M. Puszyk, 

Olorunseun Ogunwobi, Quanfeng Wu, Chen Liu 

1968 

Promoted malignant tumor phenotype of hepatitis C 

virus transgenic mice fed an atherogenic high-fat diet is 

associated with the abnormal expression of the glycolysis-related 

gene pyruvate kinase M2. 

Riuta Takabatake 1 , Masao Honda 1 , Hikari Okada 1 , Kai Takegoshi 

1 , Yoshio Sakai 1 , Taro Yamashita 1 , Naoto Nagata 3 , Toshinari 

Takamura 3 , Takuji Tanaka 2 , Shuichi Kaneko 1 ; 1 Gastroenterology, 


Kanazawa, Japan; 2 Cancer Research and Prevention, The Tohkai 

Cytopathology Institute, Gifu, Japan; 3 Department of Disease Control 

and Homeostasis, Kanazawa University Graduate School of 

Medical Sciences, Kanazawa, Japan 

Objective The relationship between hepatitis C virus (HCV)-related 

metabolic abnormalities and the progression of hepatic 

tumorigenesis remains unclear. The expression of the HCV 

open reading frame in the liver of transgenic (Tg) mice induces 

steatohepatitis, which is followed by the development of hepatocellular 

carcinoma. The atherogenic high-fat diet (Ath HFD) 

mouse model develops steatohepatitis similar to human nonalcoholic 

steatohepatitis. We investigated the expression of 

pyruvate kinase M2 (PKM2), an isoenzyme of the glycolytic 

enzyme pyruvate kinase and recently recognized as a transcription 

factor promoting cancer cell growth and the progression 

of tumorigenesis, in HCV Tg mice fed an Ath HFD 

diet. Materials and Methods Four groups of mice (n = 15–20/ 

group) were fed with a control diet or an Ath HFD diet starting 

at 8 weeks after birth for 30 and 60 weeks. The degree of 

liver steatosis, liver fibrosis, liver weight, and tumor incidence 

were measured. The expression of collagen I and IV, TGF-β, 

p-SMAD3, PKM2, Foxo1, p-Akt, Hes1, and Hey1 was evaluated 

by immunohistochemical staining, TaqMan PCR, and 

western blotting. In vitro analysis was performed by using 

PKM2-stable knockdown Huh-7 cells. Results HCV Tg mice fed 

an Ath HFD diet showed markedly promoted liver fibrosis at 

30 and 60 weeks compared with wild-type (WT) mice fed an 

Ath HFD diet. In addition, HCV Tg mice fed an Ath HFD diet 

showed markedly increased liver weight (1.5-fold) and tumor 

size (1.5-fold), but with no change in tumor frequency compared 

to WT mice. HCV Tg mice fed an Ath HFD diet showed 

a significant increase in the expression of PKM2 and fibrosis-related 

genes, such as collagen I/IV, p-smad3, and TGF-β 

together with Notch signal-related genes. HCV core protein 

was found to activate the PKM2 promoter through the induction 

of CREB. In vitro, PKM2-stable knockdown Huh-7 cells showed 

significant repression of EpCAM, AFP, Sox2, Oct4, vimentin, 

and KRT19 expression, which are markers of cancer stem cells. 

Interestingly, the addition of recombinant Notch/jagged1 to 

Huh-7 cells further activated the nuclear translocation of PKM2; 

conversely, Notch-stimulated Hes1 and Hey1 expression was 

repressed in PKM2-knockdown cells. Conclusion Promoted liver 

fibrosis and a malignant tumor phenotype were observed in 

HCV Tg mice fed an Ath HFD diet, which were associated with 

the abnormal expression of PKM2. PKM2 might be involved in 

the expression of pro-fibrotic genes and epithelial-mesenchymal 

transition signaling by activation of the Notch signaling


pathway. These findings could reveal the relationships between 

glucose metabolism, liver fibrosis, and tumorigenesis in chronic 

hepatitis C. 

Disclosures: 







The following authors have nothing to disclose: Riuta Takabatake, Masao 

Honda, Kai Takegoshi, Yoshio Sakai, Taro Yamashita, Naoto Nagata, Toshinari 

Takamura, Takuji Tanaka 

1969 

Colorectal liver metastasis is enhanced in the alcoholic 

liver: role of ethanol-mediated alterations in the liver 

microenvironment and carcinoembryonic antigen processing 

Ashley M. Mohr 4 , Robert S. Bridge 1 , Peter Thomas 3 , John Gould 2 , 

Jacy L. Kubik 1 , Carol A. Casey 2,1 , Dean J. Tuma 1 , Benita L. 

McVicker 2,1 ; 1 Internal Medicine, University of Nebraska Medical 

Center, Omaha, NE; 2 VA Nebraska-Western Iowa Health Care 

System, Omaha, NE; 3 Surgery & Biomedical Sciences, Creighton 

University, Omaha, NE; 4 Biochemistry & Molecular Biology, University 

of Nebraska Medical Center, Omaha, NE 

Colorectal cancer (CRC) is a leading cause of cancer mortality. 

The majority of CRC deaths are associated with the development 

of colorectal liver metastasis (CRLM). The characterization 

of CRLM including the role of the liver environment is ongoing 

and vital for the development of treatment options. Although it 

is known that alcohol consumption is an independent risk factor 

for CRLM, the mechanisms by which alcohol affects the environment 

influencing metastasis are undefined. It has been shown 

that the liver processing of a CRC-associated protein, carcinoembryonic 

antigen (CEA), correlates with CRLM. In particular, 

CEA is processed by Kupffer cells (KCs) and hepatocytes 

(HCs) resulting in KC-produced cytokines or the physiological 

degradation of CEA in HCs; events that respectively can potentiate 

or regulate CRLM. Interestingly, alcoholic liver disease 

(ALD) is associated with increased serum CEA levels. However, 

alcohol’s effects on CEA processing and CRLM is not known. 

In the current study, we examined the role of alcohol in a novel 

model of ALD and CRLM mediated by human CEA-expressing 

LS174T CRC cells. Methods: Immunodeficient Rag1 tm1Mom mice 

were fed control (C) or ethanol (E) diets for four weeks followed 

by intrasplenic injection of LS174T cells. The C/E diets were 

continued for 3-5 more weeks at which time ALD and CRLM 

were assessed. Results: ALD was confirmed by several parameters 

including decreased HC endocytosis via the asialoglycoprotein 

receptor (ASGPR), a well-defined early impairment in 

ALD and receptor-mediated degradation of CEA. Importantly, 

the rate and burden of CRC tumors was enhanced in the alcohol-fed 

mice. Specifically, 3 weeks following LS174T injections, 

50% of E-fed mice had CRLM compared to no evidence of 

metastasis in controls. By 5 weeks, all C/E mice had CRLM yet 

the tumor size and number were markedly increased in alcohol-affected 

livers. Analysis of liver tissue adjacent to tumors 

indicated enhanced levels of CEA and KC-associated cytokines 

(TNF-α, IL1-b, IL-10). Moreover, adhesion molecules (ICAM-1 

and E-selectin) known to be involved in CRLM in response to 

KC-produced cytokines, were elevated (2-fold) in E-fed mice. 

Overall, ALD exacerbates CRLM by potentiating CEA-mediated 

KC cytokine production and downstream adhesion molecules 

facilitating CRC adherence in the liver. Additionally, 

alcohol-mediated defects to ASGPRs led to impaired CEA degradation 

that can contribute to pro-metastatic signaling. Further 

investigation could significantly impact the understanding of 

involved mechanims as well as identify potential targets for the 

treatment of colorectal metastases in the alcohol-injured liver. 

Disclosures: 

The following authors have nothing to disclose: Ashley M. Mohr, Robert S. 

Bridge, Peter Thomas, John Gould, Jacy L. Kubik, Carol A. Casey, Dean J. Tuma, 

Benita L. McVicker 

1970 

Fibrotic liver microenvironment promotes stemness of 

liver cancer through the activation of TGF-beta and 

histone demethylase KDM2B in a p53-status dependent 

manner. 

Taro Yamashita, Mitsumasa Kondo, Hikari Okada, Naoki Oishi, 

Masao Honda, Shuichi Kaneko; Department of Gastroenterology, 



[Background] Liver cirrhosis is one of the most important risk 

factors for the development of hepatocellular carcinoma (HCC). 

Fibroblasts are known to regulate malignant nature of tumors, 

but its mechanisms remains to be elucidated in detail. In this 

study, we evaluated the role of fibroblasts on stemness of the 

tumor, which is closely associated with the aggressiveness of 

liver cancer. [Methods]Primary HCC cells obtained from surgically 

resected specimens as well as Huh7, Hep3B, and HepG2 

cells were co-cultured with various fibroblasts in vitro using cell 

culture inserts. Gene and protein expression was evaluated 

by qRT-PCR and Western blotting. Fluorescence-activated cell 

sorting (FACS) was used to evaluate the frequency of cancer 

stem cells expressing EpCAM/CD133. Cancer stem cell characteristics 

were evaluated by spheroid formation, invasion, 

and tumorigenicity in immune deficient mice. Time-lapse image 

analysis was performed to monitor cell motility affected by stromal 

cells. KDM2B and H3K36 methylation status was evaluated 

by Western blotting and immunofluorescence. [Results] Co-culture 

of HCC cells with fibroblasts resulted in the enhanced cell 

motility, spheroid formation, and invasion capacities of HCC 

cells. FACS analyses demonstrated the enrichment of EpCAM/ 

CD133-positive Huh7 cells when co-cultured with fibroblasts. 

Gene expression analysis of selected cytokines and growth factors 

indicated the abundant expression of TGFB1 in fibroblasts. 

Supplementation of recombinant transforming growth factor 

beta (TGF-beta) enhanced the spheroid formation capacity of 

HCC cells, and this effect was abolished when cultured with 

neutralizing antibodies against TGF-beta. Co-injection of HCC 

cells with fibroblasts resulted in the high frequency of lung 

metastasis. Interestingly, acquisition of stemness induced by 

TGF-beta was associated with the activation of histone demethylase 

KDM2B in p53-mutant Huh7 cells but not in p53-wildtype 

HepG2. Introduction of mutant p53 (R175H) enhanced the 

activation of KDM2B induced by TGF-beta in p53-null Hep3B 

cells. [Conclusions] Fibrotic liver microenvironment accelerates 

the malignant natures of HCC with enrichment of EpCAM/ 

CD133-positive cancer stem cells through the activation of TGFbeta 

signaling. TGF-beta may activate histone demethylase 

KDM2B to modify epigenetic status to induce the stemness of 

HCC in a p53-status dependent manner. 

Disclosures: 






Japan, Eli lilly Japan


The following authors have nothing to disclose: Taro Yamashita, Mitsumasa 

Kondo, Naoki Oishi, Masao Honda 

1971 

Development of HCC is dependent upon hepatocellular 

apoptosis in a murine model of NASH associated tumoriogenesis 

Casey Johnson 1 , Alexander Wree 1,2 , Joan Font-Burgada 3 , Akiko 

Eguchi 1 , Davide Povero 1 , Michael Karin 3 , Ariel E. Feldstein 1 ; 

1 Pediatrics, UCSD, La Jolla, CA; 2 Department of Internal Medicine 

III, University Hospital RWTH-Aachen, Aachen, Germany; 3 Laboratory 

of Gene Regulation and Signal Transduction, Departments of 

Pharmacology and Pathology, UCSD, La Jolla, CA 

Background: Nonalcoholic steatohepatitis (NASH) is increasingly 

associated with the development of hepatocellular carcinoma 

(HCC), even in patients without established liver cirrhosis. 

While the oncogenic mechanisms of blocking persistent hepatocyte 

cell death, a common feature among various chronic liver 

diseases, remain unclear it nevertheless presents as a logical 

treatment modality. Therefore, we aimed at investigating the 

influence of hepatocyte specific Bid depletion – a BH3-only 

Bcl-2 family member that amplifies apoptotic death signals – 

on tumor development in a murine model of NASH induced 

HCC. Methods: Hepatocyte-specific conditional Bid-knockout 

mice (Bid Δhep ) and functional Bid-expressing control mice (Bidflo/flo 

) were injected with 100mg/Kgr streptozotocin (STZ) at 

2-days of age. Mice were weaned onto a 60% Kcal from fat 

diet and liver tumorigenesis was investigated 5 months later. 

Results: Western Blot analysis confirmed hepatocyte specific 

Bid depletion in Bid Δhep mice. Survival rate was impaired in 

Bid flo/flo when compared to Bid Δhep mice (Bid flo/flo 3/12 vs. 

Bid Δhep 0/12; p < 0.05). Bid flo/flo mice exhibited higher incidences 

of liver tumors larger than 5mm when compared to 

Bid Δhep mice, which translated into an increased tumor load 

(Bid flo/flo 10/10 vs. Bid Δhep 4/10; p < 0.05). Liver transaminase 

levels remained normal within Bid Δhep mice, while Bid flo/ 

flo 

mice were elevated (Bid Δhep 249 IU/l ±12 IU/l (mean ± 

SEM) vs. Bid flo/flo 84±12; p < 0.05). Tissue section quantification 

revealed an increased rate of hepatocellular apoptosis 

in Bid flo/flo mice when compared to Bid Δhep mice. Quantitative 

PCR analysis of mRNA levels of AFP (alphafetoprotein), HGF 

(hepatocyte growth factor), Cyclin D1 and PCNA were significantly 

increased in Bid flo/flo mice as compared to Bid Δhep 

animals indicating cellular proliferation. Liver sections of Bid flo/ 

flo 

mice also exhibited increased infiltration of inflammatory 

cells and concomitant upregulation of mRNA levels of F4/80, 

Ly6c, TNFa (tumor necrosis factor), and IL-1beta (interleukin 

1 beta). Liver fibrosis developed in both mouse groups, while 

mRNA levels indicating hepatic stellate cell activation (aSMA 

– smooth muscle actin; and TIMP1 – tissue inhibitor of metalloproteinase) 

were significantly increased in Bid flo/flo mice when 

compared to Bid Δhep mice. Conclusion: Our study demonstrates 

that blocking hepatic apoptosis ameliorated HCC development 

in a NASH-related tumorigenesis model. These results suggest 

that reducing hepatocyte cell death, liver inflammation and 

compensatory proliferation presents a strong beneficial effect 

that supersedes the potential side effect of enhancing tumor 

cell survival. 

Disclosures: 

Akiko Eguchi - Grant/Research Support: Gilead, Conatus 

The following authors have nothing to disclose: Casey Johnson, Alexander Wree, 

Joan Font-Burgada, Davide Povero, Michael Karin, Ariel E. Feldstein 

1972 

Cytotoxic synergy between sorafenib and cyclooxygenase 

2 inhibitor celecoxib in vitro: induction of cell death 

of hepatocellular carcinoma through ER stress mediated 

apoptosis 

Sera Yang 1 , SoHee Kang 1 , Soohyun Park 2 , Jonghwa Kim 1 , Yong 

Han Paik 1,2 ; 1 Department of Medicine, Samsung Medical Center, 

Sungkyunkwan University School of Medicine, Seoul, Korea 

(the Republic of); 2 Department of Health Science and Technology, 

Samsung Advanced Institute for Health Science and Technology, 

Sungkyunkwan University, Seoul, Korea (the Republic of) 

Background/Aim: Sorafenib, a multi-kinase inhibitor, is currently 

recommended for the treatment of advanced hepatocellular 

carcinoma (HCC). However, the response rate of sorafenib 

in advanced HCC treatment are not satisfactory and there is 

a rationale for investigating its use in combination with other 

agents. Celecoxib is known as a selective cyclooxygenase-2 

(COX-2) inhibitor and has been known to exhibit anti-tumor 

effects in HCC cells but the mechanism is still largely unknown. 

The aim of this study was to investigate the synergistic effect of 

celecoxib on the anti-tumor activity of sorafenib in HCC and 

to determine the underlying molecular mechanism. Methods: 

HCC cell line PLC/PRF/5 were treated with sorafenib and/ 

or celecoxib, and then proliferation was analyzed by MTS 

assay. Cell death was evaluated by annexin staining and 

FACS analysis. Western blot was also used to study the mechanism 

of the induced cell death. Results: MTS assay revealed 

that each of sorafenib and celecoxib alone marginally inhibited 

HCC cell proliferation. However, the combination of 

sorafenib and celecoxib showed a synergistic effect to inhibit 

cell proliferation, while the effect was less strongly observed 

with 2,5-dimethyl-celecoxib (DMC, a derivative of celecoxib 

that lacks COX2-inhibitory function). Using flow cytometry, we 

found that the combined treatment synergistically increased the 

population of Annexin + /PI + at 48h, indicating the increased 

apoptosis. The increased cleavage of caspase-8 and PARP was 

also demonstrated in Western blot. In addition, we also found 

that ER stress marker proteins, such as GRP78/Bip, CHOP, 

and spliced XBP-1, were dramatically increased in the combined 

treatment of sorafenib and celecoxib. The activation of 

caspases and cell death induced by sorafenib plus celecoxib 

was significantly inhibited by pretreatment with 4μ8C, a ER 

stress inhibitor. These data indicated that sorafenib and celecoxib 

synergistically induce cell death of HCC cell line through 

ER-stress-mediated apoptosis. Conclusion: Combined treatment 

of sorafenib and celecoxib synergistically inhibit the growth 

of HCC cells via inducing ER-stress mediated apoptosis. This 

finding raises the possibility of the combined treatment using 

sorafenib and celecoxib for the augmentation of anticancer 

effect of sorafenib in HCC. 

Disclosures: 

The following authors have nothing to disclose: Sera Yang, SoHee Kang, 

Soohyun Park, Jonghwa Kim, Yong Han Paik


1973 

PPPDE1 is a deubiquitinase that promotes HCC development 

by stabilizing a 25kd MDM2 N terminal fragment 

and subsequently suppressing the p53 pathway 

Xing-Wang Xie 1,2 , Yang-Jing Zhao 1,2 , Xue-Yan Wang 1,2 , Ran 

Fei 1,2 , Heng-Hui Zhang 1,2 , Wei-Jia Liao 3 , Lai Wei 1,2 , Yu Wang 4 , 

Hong-Song Chen 1,2 ; 1 Peking University People’s Hospital, Peking 

University Hepatology Institute, Beijing, China; 2 Beijing Key Laboratory 

of Hepatitis C and Immunotherapy for Liver Diseases, Beijing, 

China; 3 Guilin Medical College, Guilin, China; 4 Chinese 

Center for Disease Control and Prevention, Beijing, China 

Introduction: In our previous study, PPPDE1 (DESI2) was identified 

and validated as a HCC driver gene, but the underlying 

mechanism was unclear. The PPPDE1 protein belongs to a protein 

family named PPPDE (Permuted Papain fold Peptidases of 

Ds-RNA viruses and Eukaryotes) that was predicted to be a 

deubiquitinating enzyme (DUB) family by protein homology 

search. Hence, this study aims to validate the DUB activity of 

PPPDE1 and to elucidate how it is involved in the HCC development. 

Method: Lentvirus expressing wild type PPPDE1 and 

PPPDE1 C108S mutant (the cysteine 108 was replaced by serine) 

were used for over-expression and protein purification. Lentivirus 

expressing PPPDE1 specific shRNAs were used to silencing 

PPPDE1. The DUB activity of PPPDE1 and PPPDE1 C108S 

were determind by in vitro and in vivo de-ubiquitination assay. 

Co-immunoprecipitation (Co-IP) was used in combination with 

Mass Spectrometry and Western blot to identify and validation 

protein binding partners of PPPDE1. Colony formation 

assay and subcutaneous tumor model were used to evaluate 

the clonogenic and tumorigenic ability of HCC cells. Results: 

Purified PPPDE1 protein cleaved K48-linked and K63-linked 

Tetra-Ubiquitin into monoubiquitin in vitro. Transfected PPPDE1 

also markedly reduced the K48-linked and K63-linked protein 

ubiqutination level in cultured cells. These results showed that 

PPPDE1 is a novel DUB and the PPPDE protein family is a new 

DUBs family with cysteine proteases activity. The C108S substitution 

of PPPDE1 C108S destroyed the deubiquitination activity 

of PPPDE1. Moreover, PPPDE1 C108S lost the ability to promote 

the clonogenic growth of HCC cell lines compared to wild 

type PPPDE1, which suggested that PPPDE1 promote the HCC 

development mainly through its DUB activity. We also found 

that PPPDE1 stabilized a 25kd N termal MDM2 protein fragment 

(MDM2 p25), a degradation fragment from MDM2-FL 

(full length) and MDM2 p60 (60kd) protein, by binding and 

deubiquitinated this protein fragment. When over-expressed 

in HCC cell lines, PPPDE1 significantly promoted the degradation 

of p53 through a MDM2 p25 dependent manner and 

down regulated the protein level of BAX, a downstream effector 

of p53-induced apoptosis. Reversely, knockdown of PPPDE1 

in HCC cell lines considerably increased the p53 and BAX 

protein level while greatly down-regulated the MDM2 p25 

level. Most importantly, PPPDE1 silencing with lentiviral shRNA 

tremendously inhibited the in vivo growth of subcutaneous 

xenografts tumor of HCC cell lines in nude mice. Conclusion: 

PPPDE1 is a novel DUB that can promote the development of 

HCC by stabilizing a 25kd N terminal MDM2 fragment and 

subsequently suppressing the p53 pathway. 

Disclosures: 



The following authors have nothing to disclose: Xing-Wang Xie, Yang-Jing Zhao, 

Xue-Yan Wang, Ran Fei, Heng-Hui Zhang, Wei-Jia Liao, Yu Wang, Hong-Song 

Chen 

1974 

Mevalonate pathway targets FoxM1 transcription factor 

via protein geranylgeranylation in human hepatocellular 

carcinoma 

Satoshi Ogura, Yuichi Yoshida, Mayumi Egawa, Tomohide Kurahashi, 

Kunimaro Furuta, Shinichi Kiso, Yoshihiro Kamada, Tetsuo 

Takehara; Department of Gastroenterology and Hepatology, 

Osaka University, Suita, Osaka, Japan 

Background & Aims: Mevalonate (MV) pathway, which is 

required for cholesterol biosynthesis, has been implicated in 

the pathogenesis of hepatocellular carcinoma (HCC). Inhibition 

of HMG-CoA reductase (HMGCR), a rate limiting enzyme for 

MV pathway, has been also reported to reduce incidence of 

HCC in animal and clinical studies. The Forkhead box M1 

(FoxM1) is a proliferation-specific transcription factor and is 

over-expressed in a variety of cancers, including HCC. In addition, 

loss of FoxM1 is shown to reduce hepatocarcinogenesis 

in mouse models. However, to date, it remains unclear whether 

FoxM1 is involved in MV pathway of HCC. In this study, we 

aimed to elucidate this issue using in vitro culture systems. 

Method: Human hepatoma cell lines HepG2, Huh7, and HLF, 

were treated with chemical inhibitors of enzymes in MV pathway 

for 24 hours and the effect of these inhibitors on FoxM1 

protein expression was examined by Western blot analysis. 

Results: Administration of statin (pitavastatin), HMGCR inhibitor, 

induced an about 50 % reduction of FoxM1 expression 

in hepatoma cells (p


1975 

Retrospective cohort study for liver carcinogenesis prediction 

among HBs antigen negative and anti-HCV antibody 

negative hepatitis patients 

Tomoko Aoki 2,1 , Hiroko Iijima 1 , Takashi Nishimura 1 , Chikage 

Nakano 1 , Tomoyuki Takashima 1 , Nobuhiro Aizawa 1 , Naoto 

Ikeda 1 , Hironori Tanaka 1 , Yoshinori Iwata 1 , Hirayuki Enomoto 1 , 

Masaki Saito 1 , Shuhei Nishiguchi 1 ; 1 Hyogo College of Medicine, 

Nishinomiya city, Hyogo prifecture, Japan; 2 Internal medicine, 

YOKA HOSPITAL, Yoka 1878-1, Japan 

BACKGROUND & AIMS:Recently, we can use new ultrasonographic 

methods for non-invasive diagnosis of liver fibrosis. 

The aim for this study is to reveal the risk of liver carcinogenesis 

among HBs antigen negative and anti-HCV negative 

(nonBnonC) cases using Virtual Touch Quantification (VTQ). 

METHODS:Our research model was a retrospective cohort 

study. We continued to follow up 910 patients for 45.2 

months: 23 patients developed into hepatocellular carcinoma 

(HCC), we named them “carcinogenesis group”. The other 

887 patients did never develop HCC, we named them “carcinogenesis-free 

group”. We examined factors that contribute 

to carcinogenesis by Cox regression analysis. RESULT:The univariate 

analysis between two groups: there were significant difference 

in age, sex, platelet counts, Hyaluronic Acid (HA), FIB4 

index, VTQ, PT-INR, serum albumin, fasting plasma glucose 

(FPG), and total cholesterol(p


expression of keratin 19(K19), epithelial cell adhesion molecule(EpCAM), 

matrix metalloproteinase 2/9(MMP2/9) and 

CXC chemokine receptor 4(CXCR4) were also down-regulated 

in Bel 7402 cells; Migration and invasion, expression of K19, 

EpCAM, MMP2/9 and CXCR4 were significant enhanced 

when HLE cells(non-AFP-producing) were transfected with AFP 

expressed vector. Conclusions: AFP plays a critical role in promoting 

metastasis of HCC; AFP is significantly promote HCC 

cell invasion and metastasis via up-regulated expression of 

metastasis-related proteins. Thus, AFP may be used as a novel 

therapeutic target for treating HCC patients. 

Disclosures: 

The following authors have nothing to disclose: Mingyue Zhu, Junli Guo, Wei 

Li, Mengsen Li 

1978 

Osteopontin-c isoform promotes a mesenchymal phenotype 

in human cholangiocarcinoma cells 

Marco A. Briones-Orta 1 , Jason D. Coombes 1 , Massimiliano Mellone 

7 , Rossella Rispoli 5 , Paul P. Manka 1,3 , Rasha Younis 1 , Naoto 

Kitamura 1 , Shannon S. Glaser 6,8 , Gianfranco Alpini 6,8 , Alberto 

Quaglia 9 , Roger Williams 1 , Salvatore Papa 4 , Ali Canbay 3 , Wing- 

Kin Syn 1,2 ; 1 Regeneration and Repair, The Institute of Hepatology, 

London, United Kingdom; 2 Liver Unit, Barts Health NHS Trust, London, 

United Kingdom; 3 Gastroenterology and Hepatology, University 

Hospital Essen, Essen, Germany; 4 Cell Signaling, The Institute 

of Hepatology, London, United Kingdom; 5 Weatherall Institute of 

Molecular Medicine, John Radcliffe Hospital, Oxford, United Kingdom; 

6 Research, Central Texas Veterans Health Care Systems, 

Temple, TX; 7 Experimental Pathology Group, Cancer Sciences 

Unit, Southampton General Hospital, Southampton, United Kingdom; 

8 Scott & White Digestive Disease Research Center, Texas 

A&M Health Science Center College of Medicine, Temple, TX; 

9 Liver Pathology, Kings College London, London, United Kingdom 

Introduction: Osteopontin (OPN) is upregulated in tissue fibrosis 

and cancer. The OPN gene generates 3 isoforms (a, b and 

c) by alternative splicing, which can induce differing cellular 

responses in tumorigenesis. Although these isoforms have been 

investigated in breast, prostate, and ovarian cancers, studies in 

cholangiocarcinoma (CCA) have been limited to OPN-a (total 

OPN). We hypothesized that OPN isoforms are overexpressed 

in CCA, and the pattern of isoform overexpression is associated 

with either a more or less aggressive phenotype. Crosstalk 

between OPN and TGF-b signaling was evaluated as both 

are key modulators of cancer progression. Methods: HuCCT1, 

SG231 and CCLP1, human intrahepatic cholangiocarcinoma 

cell lines were used. Plasmids for each OPN isoform were used 

for overexpression. Expression of OPN-a, b, c, and epithelial-mesenchymal 

transition (EMT) markers (vimentin, aSMA, 

E-cadherin) were evaluated by qRT-PCR. Global gene expression 

changes were examined by microarray using SG231. 

OPN isoform overexpression and their effects on the components 

of TGF-b pathway were evaluated by immunoblots. To 

evaluate the in vivo significance of the individual isoforms, a 

xenograft model was used. FFPE human cholangiocarcinoma 

and healthy liver sections were stained for total OPN. Results: 

Total OPN was upregulated in cholangiocarcinoma. CCLP1 

cells expressed the highest levels of total OPN and exhibited 

the most mesenchymal phenotype (high vimentin and low 

E-cadherin). In contrast, HuCCT1 expressed the lowest amount 

of OPN and exhibited an epithelial phenotype (high E-cadherin 

and low vimentin). In all 3 cell lines, OPN-a, and b mRNA 

were more abundant than OPN-c (~10 fold). Overexpression 

of OPN-c led to the greatest increase in mesenchymal genes 

and proteins (vimentin and aSMA), and was associated with 

the highest level of TGF-β signalling as shown by Smad2/3 

phosphorylation. Microarray confirmed that overexpression 

of OPN-c induced the largest alterations in gene expression 

profile, and an enrichment analysis revealed that OPN modulated 

genes which regulate phosphoprotein and acetylation. In 

the xenograft model, overexpression of all OPN isoforms was 

associated with greater rates of tumour growth than control, 

but OPN-c had the greatest rate. Conclusions: OPN expression 

correlates with the degree of EMT (marker of aggressiveness) 

in human cholangiocarcinoma. OPN-c is associated with the 

most mesenchymal phenotype and TGF-β pathway activation. 

OPN-c also induces greater changes in the transcriptome and 

supports greater rates of tumor growth than OPN-a, and -b. 

High OPN-c expression may be a predictor of worse clinical 

outcomes. 

Disclosures: 

The following authors have nothing to disclose: Marco A. Briones-Orta, Jason D. 

Coombes, Massimiliano Mellone, Rossella Rispoli, Paul P. Manka, Rasha Younis, 

Naoto Kitamura, Shannon S. Glaser, Gianfranco Alpini, Alberto Quaglia, Roger 

Williams, Salvatore Papa, Ali Canbay, Wing-Kin Syn 

1979 

Glypican 3 (GPC3)-CD81 axis in regulation Hippo pathway 

in hepatocytes and hepatocellular carcinoma (HCC) 

Yuhua Xue, Kelly Koral, William C. Bowen, Anne Orr, Meagan 

Haynes, Wendy M. Mars, George K. Michalopoulos; Pathology, 


GPC3 is over-expressed in HCC. Mutations of GPC3 in humans 

are the cause of Simpson-Golabi-Behmel syndrome, associated 

to organomegaly. Previous studies have shown that GPC3 is 

associated with inhibition of hepatocyte growth and GPC3 

transgenic mice with targeted expression in hepatocytes have 

decreased liver regeneration. We have also shown that GPC3 

binds to the membrane tetraspanin CD81, which is one of the 

portals of entry of hepatitis C virus (HCV). Studies elsewhere 

have shown that activation of CD81 is associated with phosphorylation 

of Ezrin. The latter has been shown to regulate 

the Hippo pathway, responsible for regulating nuclear levels 

of the protein known as Yes-associated protein (Yap). Expression 

of Yap in the nucleus relates to activation of hepatocyte 

growth. Mice over-expressing GPC3 have decreased levels of 

nuclear Yap. Since we have demonstrated that GPC3 binds 

to CD81, the purpose of this study is to explore the role of 

GPC3 in regulation of Yap via CD81 and Hippo pathway and 

to investigate the possible mechanisms. We found that treatment 

of primary rat hepatocytes with CD81 agonist antibody 

led to higher levels of phosphorylated Ezrin and lower Hippo 

activity. Partial hepatectomy (PHx) exhibited that loss of GPC3 

antagonistic modulation during early liver regeneration, CD81 

activated Ezrin and down regulated Hippo activity in vivo. Rat 

hepatoma cell lines JM1 and JM2 had dramatically down regulated 

CD81 expression and Hippo activity, and up regulated 

Ezrin activity in vitro. Re-gain of CD81 expression in JM2 rat 

hepatoma cells up regulated Hippo activity while down regulated 

Ezrin activity in vitro. Human HCC tissue array revealed 

absence of CD81 in most HCCs but present in all the normal 

tissue controls, while GPC3 was present in most HCCs and all 

the normal tissue controls. Treatment with HCV envelope protein 

E2 led to more Hippo activity and decrease in nuclear Yap 

in HepaRG cells and primary human hepatocytes. Conclusions: 

The CD81-phosphoEzrin signaling enhances nuclear Yap levels. 

This pathway in normal hepatocytes is inhibited by GPC3. 

In human hepatocytes, GPC3 and the E2 protein of HCV also 

inhibit the CD81-phosphoEzrin effect on Hippo pathway and 

cause a decrease in nuclear Yap. Since most HCC lack expression 

of CD81, they are likely to escape from antagonistic mod-


ulation of over-expressed GPC3. HCC may also have a growth 

advantage over normal hepatocytes in HCV infection by missing 

CD81, thus being able to undergo clonal expansion, facilitating 

HCC early development and growth. Persistent HCV 

infection strengthens this expansion process and contributes 

carcinogenesis by E2 protein interacting with CD81. 

Disclosures: 

George K. Michalopoulos - Consulting: Vital Therapies 

The following authors have nothing to disclose: Yuhua Xue, Kelly Koral, William 

C. Bowen, Anne Orr, Meagan Haynes, Wendy M. Mars 

gene that acts as a driving force for the amplification at 3q26 

in HCC and that the oncoprotein EVI1 suppresses TGF-β-mediated 

growth inhibition of HCC cells. 

Disclosures: 






The following authors have nothing to disclose: Kohichiroh Yasui, Atsushi 

Umemura, Taichiro Nishikawa, Kanji Yamaguchi, Michihisa Moriguchi, Yoshio 

Sumida, Hironori Mitsuyoshi, Shinji Tanaka, Shigeki Arii 

1980 

EVI1 is a target for gene amplification at 3q26 and 

suppresses growth inhibition by transforming growth 

factor-β in hepatocellular carcinoma 

Kohichiroh Yasui 1 , Atsushi Umemura 1 , Taichiro Nishikawa 1 , Kanji 

Yamaguchi 1 , Michihisa Moriguchi 1 , Yoshio Sumida 1 , Hironori Mitsuyoshi 

1 , Shinji Tanaka 2 , Shigeki Arii 3,4 , Yoshito Itoh 1 ; 1 Department 

of Molecular Gastroenterology and Hepatology, Kyoto 

Prefectural University of Medicine, Kyoto, Japan; 2 Department 

of Molecular Oncology, Tokyo Medical and Dental University, 

Tokyo, Japan; 3 Hamamatsu Rosai Hospital, Hamamatsu, Japan; 

4 Department of Hepato-Biliary-Pancreatic Surgery, Tokyo Medical 

and Dental University, Tokyo, Japan 

Background: Amplification of DNA in certain regions of chromosomes 

plays a crucial role in the development and progression 

of human malignancies, specifically when proto-oncogenic 

target genes within those amplicons are overexpressed. A common 

criterion for designation of a gene as a putative target 

of amplification is that gene amplification leads to its overexpression. 

The EVI1 (ecotropic viral integration site 1) gene 

codes for a zinc finger transcriptional factor that is involved in 

leukemic transformation of hematopoietic cells. EVI1 is one of 

the most aggressive oncogenes associated with myeloid leukemia. 

EVI1 has been reported to suppress transforming growth 

factor (TGF)-β signaling by inhibiting Smad3 in leukemic cells. 

However, little is known about its relevance for hepatocellular 

carcinoma (HCC). Aim: To identify a novel gene that acts as a 

driving force for gene amplification in HCC and is involved in 

the development and progression of HCC. Methods: We investigated 

DNA copy number aberrations in 20 HCC cell lines 

using a high-density oligonucleotide microarray (GeneChip 

Mapping 100K and 250K arrays, Affymetrix). DNA copy 

number and expression of EVI1 were determined by using fluorescence 

in situ hybridization (FISH) and quantitative PCR in 

HCC cell lines and primary HCCs. Knockdown experiments of 

EVI1 were performed. Results: We found that a novel amplification 

at the chromosomal region 3q26 occurs in the HCC cell 

line, JHH-1, and that MDS1 (myelodysplastic syndrome 1) and 

EVI1 complex locus (MECOM ), which lies within the 3q26 

region, was amplified. Quantitative PCR analysis of the three 

transcripts transcribed from MECOM indicated that only EVI1, 

but not the fusion transcript MDS1-EVI1 or MDS1, was overexpressed 

in JHH-1 cells and was significantly up-regulated in 22 

(61%) of 36 primary HCC tumors when compared with their 

non-tumorous counterparts. A copy number gain of EVI1 was 

observed in 24 (36%) of 66 primary HCC tumors. High EVI1 

expression was significantly associated with larger tumor size 

and higher level of des-γ-carboxy prothrombin (DCP), a tumor 

marker for HCC. Knockdown of EVI1 resulted in increased 

induction of the cyclin-dependent kinase inhibitor p15 INK4B by 

TGF-β and decreased expression of c-Myc, cyclin D1, and 

phosphorylated Rb in TGF-β-treated cells. Consequently, knockdown 

of EVI1 led to reduced DNA synthesis and cell viability. 

Conclusions: Our results suggest that EVI1 is a probable target 

1981 

Hyperinsulinemia, not hyperglycemia accelerates the 

progression of hepatocellular carcinoma in neonatal 

streptozotocin induced mouse model 

Koichi Tsuneyama, Ryosuke Bessho, Masahiro Miki, Hayato Baba, 

Tetsuyuki Takahashi, Hirohisa Ogawa, Hisanori Uehara; Department 

of Pathology and Laboratory Medicine, Institute of Biomedical 

Sciences, Tokushima University Graduate School, Tokushima, 

Japan 

Purpose: Diabetes mellitus (DM) is a well-known risk factor 

for hepatocellular carcinoma (HCC); however, the underlying 

mechanisms are not well understood. We have previously 

reported that neonatal streptozotocin (STZ) treatment in DIAR 

mice (DIAR-nSTZ) causes type 1 diabetes and subsequent HCC 

without special diet. In the following study, to examine the 

relation between hyperglycemia and HCC, we normalized the 

blood glucose level by applying excess insulin in DIAR-nSTZ 

mice. Although diabetic features were completely improved, 

frequency of HCC did not improve in 12 weeks of age. We 

hypothesized that hyperinsulinemia, not hyperglycemia have 

a close association to carcinogenesis in this model. To clarify 

this hypothesis, we examined neonatal STZ treatment in IV CS 

mice, which showed quick normalization of blood glucose level 

after STZ treatment by rapid regeneration of islet cells. Methods: 

Newborn male IV-CS mice were prepared at institute of 

animal reproduction (Ibaraki, Japan). STZ (60 mg/g) was subcutaneously 

injected into the IV CS-STZ treated group (IV-CSnSTZ 

mice, N13), whereas physiologic solution was injected 

into the control group (IV-CS-control mice, N=13) at 1.5 days 

after birth. All mice were fed a normal diet, and physiologically 

and histopathologically assessed at 8 (N=2), 12 (N=3) and 

16 (N-8) weeks of age. Blood was taken once in month and 

measured glucose and insulin level in all mice. Results: Blood 

glucose level of IV CS-nSTZ mice showed slightly increase till 

5 week of age, but it was normalized after 6 weeks of age. In 

contrast, blood insulin level showed higher titer after 6 weeks 

of age. Even at 8 weeks of age, one out of two mice showed 

hepatic nodule in a diameter of 2mm. Histopathologically, it 

showed mild structural and nuclear atypia showing dysplastic 

nodule. At 12 weeks of age, one out of three mice contained 

2 hepatic nodules of dysplastic nodule. At 16 weeks of age, 

six out of eight mice showed one or more hepatic nodules. 

Histologically, well differentiated hepatocellular carcinoma 

as well as dysplastic nodules were observed. In contrast, no 

hepatic nodules were recognized in IV CS-control mice. Conclusions: 

IV CS mice showed hepatocarcinogenesis after neonatal 

STZ treatment. Differently from other strains including ddY 

and DIAR, blood glucose level did not show marked increase 

after STZ treatment, and it was normalized after 6 weeks of 

age. However, frequency of HCC showed similar tendency 

of DIAR-nSTZ mice. In addition to our previous data of insulin 

treatment of DIAR-nSTZ mice, we strongly hypothesized that


hyperinsulinaemia rather than hyperglycemia can accelerate 

the progression of HCC. 

Disclosures: 

The following authors have nothing to disclose: Koichi Tsuneyama, Ryosuke 

Bessho, Masahiro Miki, Hayato Baba, Tetsuyuki Takahashi, Hirohisa Ogawa, 

Hisanori Uehara 





The following authors have nothing to disclose: Yoshiki Onishi, Tomohide Tatsumi, 

Satoshi Aono, Seiichi Tawara, Akira Nishio, Atsuo Takigawa, Takahiro 

Suda, Tadashi Kegasawa, Shigeki Suemura, Minoru Shigekawa, Ryotaro Sakamori, 

Naoki Hiramatsu 

1982 

Branched chain amino acids promote sorafenib-mediated 

apoptosis in hepatocellular carcinoma via 

down-regulation of Mcl-1 

Yoshiki Onishi, Tomohide Tatsumi, Satoshi Aono, Seiichi Tawara, 

Akira Nishio, Atsuo Takigawa, Takahiro Suda, Tadashi Kegasawa, 

Shigeki Suemura, Minoru Shigekawa, Hayato Hikita, 

Ryotaro Sakamori, Naoki Hiramatsu, Tetsuo Takehara; Department 

of Gastroenterology and Hepatology, Osaka University Gradute 


Background&aim: Sorafenib is an only effective chemotherapeutic 

agent in the treatment of hepatocellular carcinoma 

(HCC). However, the survival benefit is limited. Branched 

chain amino acids (BCAAs) promote albumin synthesis through 

the activation of mTOR pathway. BCAAs treatment has been 

reported to reduce the incidence of HCC in patients with liver 

cirrhosis and to reduce the recurrence of HCC after curative 

treatment. We previously reported a retrospective study that 

the combined use of sorafenib and BCAAs prolonged the 

overall survival in HCC patients. In this study, we examined 

the mechanism of anticancer effect of the combination therapy. 

Method: Human HCC cell lines, HepG2, PLC/PRF/5 and 

Huh7, were treated with sorafenib and BCAAs. Cell viabiliy 

was evaluated by WST assay. To evaluate apoptosis of HCC 

cells, we measured caspase3/7 activities in the culture supernatants 

and Annexin-V positive cells by flow cytometry. Growth 

signals and regulatory proteins of apoptosis were analyzed 

by western blotting. mRNA levels were evaluated by using 

quantitative RT-PCR. Result: The combined use of sorafenib and 

BCAAs significantly decreases in cell viability of all treated 

HCC cells. Western blotting analysis revealed that addition 

of BCAAs enhanced inhibition of ERK pathway in sorafenibtreated 

HCC cells. Sorafenib treatment induced Akt phosphorylation 

in HCC cells as previously reported. However, addition 

of BCAAs antagonized the Akt phosphorylation. The combined 

use of sorafenib and BCAAs resulted in significant increase 

in caspase 3/7 activities and Annexin-V positive cell rates in 

HCC cells compared with sorafenib alone. We next examined 

pro-and anti-apoptotic proteins in HCC cells treated with 

sorafenib and BCAAs. Addition of BCAAs enhanced reduction 

of anti-apoptotic protein Mcl-1 expression in sorafeib-treated 

HCC cells. In contrast, both pro-apoptotic proteins Bak/Bax 

and anti-apoptotic protein Bcl-xL did not changed. mRNA 

expression levels of Mcl-1 were significantly lower in HCC cells 

treated by sorafenib with BCAAs than those without BCAAs. 

Furthermore, we evaluated Mcl-1 degradation in the HCC cells 

by using cycloheximide, a protein synthesis inhibitor. After inhibition 

of protein synthesis, Mcl-1 levels were decreased more in 

sorafenib-treated cells with BCAAs than those without BCAAs. 

These results indicate that BCAAs reduce Mcl-1 expression at 

both transcriptional and post-transcriptional levels in sorafenibtreated 

HCC cells. Conclusion: BCAAs enhanced sorafenib-mediated 

apoptosis in HCC cells via down regulation of Mcl-1. 

These results suggest that the combined use of sorafenib and 

BCAAs is effective in the treatment of advanced HCC patients. 

Disclosures: 


1983 

DNA methylome and cancer-specific expression change 

of clustered miRNAs in non-B non-C hepatocellular carcinoma 

Takeshi Matsui 2,1 , Masanori Nojima 8 , Etsuko Iio 2 , Akihiro Tamori 3 , 

Shoji Kubo 4 , Ken Shirabe 5 , Koichi Kimura 9 , Mitsuo Shimada 9 , 

Tohru Utsunomiya 6 , Yasuteru Kondo 7 , Yasuhito Tanaka 2 ; 1 Center 

for Gastroenterology, Teine-Keijinkai Hospital, Sapporo, Japan; 

2 Department of Virology unit, Nagoya City University Graduate 

School of Medical Sciences, Nagoya, Japan; 3 Department of 

Hepatology, Osaka City University Graduate School of Medicine, 

Osaka, Japan; 4 Department of Hepato-Biliary-Pancreatic Surgery, 

Graduate School of Medicine, Osaka City University, Osaka, 

Japan; 5 Department of Surgery and Science, Graduate School 

of Medical Sciences, Kyushu University, Fukuoka, Japan; 6 Oita 

prefectual hospital, Oita, Japan; 7 Division of Gastroenterology, 

Tohoku University Hospital, Sendai, Japan; 8 Institute of Medical 

Science Hospital, the University of Tokyo, Tokyo, Japan; 9 Department 

of Surgery, the University of Tokushima, Tokushima, Japan 

[Background] While HBV and HCV infection have been suppressed 

with development of public health measures and 

medical treatment, non-B non-C hepatocellular carcinoma 

(NBNC-HCC) is considered increasing based on increase of 

nonalcoholic fatty liver disease (NAFLD) in Japan. Molecular 

biological mechanism should be pursued against this disease 

structure change. [Aim] To explore critical changes of DNA 

methylation in micro RNA (miRNA) coding regions and its 

expression in tumorigenesis of NBNC-HCC. [Methods] Infinium 

HumanMethylation450 BeadChip Kit (Illumina) and micro 

3D-Gene miRNA Oligo Chip (Toray) were applied in 26 pairs 

of tumor and non-tumor background tissue samples. Pooled 

analysis using the data from Gene Expression Omnibus (GEO, 

NCBI) was also conducted to strengthen generalizability, and 

to seek common DNA methylation changes across infection 

status (HBV, HCV or NBNC). Gene ontology analysis were 

applied for gene clusters defined by clustering and other statistical 

characteristics. [Results and plan] Hierarchical clustering 

of the DNA methylome status demonstrates clear discrimination 

between tumor and non-tumor samples. While approx. 20% of 

CpG islands in promoters were dominantly hypermethylated 

in the tumor, other regions were globally hypomethylated in 

the tumor. The mean difference in methylation levels of total 

probes was -10.5% in non-CpG islands, and 0.68% in CpG 

islands (tumor vs. non-tumor). It suggests that the change of 

methylation status was totally different between CpG and non- 

CpG islands. Next, to investigate the molecular biological significance 

of DNA hypomethylation in the tumor, we determine 

the characteristics of hypomethylated sites, and then found several 

miRNA clusters were markedly hypomethylated (-30% to 

-40%) in tumor tissues (e.g. miR-493 in 14q32.2, miR-154 in 

14q32.31, miR-518F in 19q13.41, miR-888 in Xq27.3). Average 

methylation level in miRNA coding region were lower than 

other regions. In detail, 52.8% of total miRNA probes were significantly 

hypomethylated, and mean difference in methylation 

levels were -14.4% in non-CpG islands, and 0.14% in CpG 

islands. Consistent with the hypomethylation tendency, average 

miRNA expression level was 28.0% up-regulated. In particular, 

miRNA expression significantly around 80% to 100%


up-regurated in the above hypomethylated miRNA clusters. 

[Conclusions] Several major miRNA clusters were markedly 

hypomethylated in tumor tissues, and expression of miRNAs 

coded in clustered-regions were broadly up-regulated. We will 

present the results of further advanced analyses including the 

influence of hypermethylation for target genes, and integration 

of clinical information. 

Disclosures: 



Squibb 

The following authors have nothing to disclose: Takeshi Matsui, Masanori 

Nojima, Etsuko Iio, Akihiro Tamori, Shoji Kubo, Ken Shirabe, Koichi Kimura, 

Mitsuo Shimada, Tohru Utsunomiya, Yasuteru Kondo 

1984 

The analyses of oxidative DNA repair genes and hepatocarcinogenesis 

in patients with chronic hepatitis C 

Koji Miyanishi, Masayoshi Kobune, Shingo Tanaka, Toshifumi 

Hoki, Tsutomu Sato, Yasushi Sato, Rishu Takimoto, Junji Kato; 

Medical Oncology and Hematology, Sapporo Medical University, 

Sapporo, Japan 

Background and aim: Recent studies have shown that excess 

hepatic iron accumulation contributes to liver injury in patients 

with chronic hepatitis C (CHC). Free iron in the liver is believed 

to facilitate the formation of reactive oxygen species (ROS), 

including hydroxyl (OH) radicals, which cause oxidative damage 

of numerous cellular components such as nucleic acids. 

The OH radical is known to generate promutagenic bases such 

as 8-hydroxy-2-deoxyguanosine (8-OHdG), which has been 

implicated in DNA mutagenesis and carcinogenesis. 8-OHdG 

lesions may be promptly repaired by human 8-oxo-guanine 

DNA glycosylase (hOGG1) and human MUTY homolog 

(MUTYH). Therefore, it is plausible that the development of 

hepatocellular carcinoma (HCC) from CHC is determined by 

the balance between DNA damage and repair. In this study, 

we conducted analyses of single nucleotide polymorphisms 

(SNPs) of hOGG1 and MUTYH genes in CHC patients with 

or without development of HCC. Further, we examined the 

efficacy of N-acetyl cysteine (NAC) as a treatment to prevent 

the development of HCC using MUYTH null mice. Methods: 

Genomic DNA and total RNA were extracted from peripheral-blood 

mononuclear cells obtained from 63 CHC patients with 

(HCC group, n=39) or without HCC (non-HCC group, n=24). 

We analyzed the SNPs of hOGG1 and MUTYH by target 

resequence using next generation sequencer (ION PGM) and 

mRNA of hOGG1 and MUTYH by Taqman RT-PCR. The HCC 

incidence rate was calculated based on the period between 

the diagnosis of CHC and the appearance of HCC, using the 

Kaplan-Meier technique. In vivo study, male C57BL/6 MUTYH 

null mice were fed an excess-iron diet or control diet with or 

without NAC. Results: The frequency of SNPs in hOGG1 did 

not differ between HCC group and non-HCC group. In one 

of MUTYH SNPs (rs3219487), the frequency of minor allele 

carrier in the HCC group significantly higher than that in HCC 

group. In addition, the expression of MUTYH mRNA in minor 

homo allele were lower than that in major homo allele. Multivariate 

Cox regression analysis indicated that age, liver fibrosis 

factor and the minor allele carrier independently associated 

with hepatocarcinogenesis. Hepatocarcinogenesis rate in 

MUTYH null mice which were fed an excess-iron diet were significantly 

higher than that in wild type C57BL/6 or in null mice 

which were fed a control diet. NAC administration decreased 

the development of HCC in null mice. Conclusions: These results 

indicated that the minor allele of this SNP in MUTYH could be 

a significant risk factor of liver carcinogenesis and could be 

a predictive marker of HCC development in CHC patients. 

Further, NAC may be useful for prevention of HCC in such 

patients. 

Disclosures: 

The following authors have nothing to disclose: Koji Miyanishi, Masayoshi 

Kobune, Shingo Tanaka, Toshifumi Hoki, Tsutomu Sato, Yasushi Sato, Rishu 

Takimoto, Junji Kato 

1985 

Continuous hepatocyte apoptosis accelerates diethylnitrosamine-induced 

tumor development in the liver 

Yasutoshi Nozaki, Hayato Hikita, Satoshi Tanaka, Sadatsugu 

Sakane, Yugo Kai, Yuki Makino, Tasuku Nakabori, Yoshinobu 

Saito, Ryotaro Sakamori, Naoki Hiramatsu, Tomohide Tatsumi, 

Tetsuo Takehara; Osaka University Graduate School of Medicine, 

Suita, Osaka, Japan 

Background and Aim: Apoptosis serves as an important mechanism 

for removing DNA damaged-cells and is considered to 

inhibit carcinogenesis. On the other hand, hepatocyte apoptosis 

is a key feature of chronic liver disease including viral hepatitis 

and steatohepatitis, which are well-known risk factors for 

HCC. Thus, it remains unclear whether the apoptosis inhibition 

accelerates or decelerates liver tumor development. The present 

study examined the impact of continuous hepatocyte apoptosis 

and inhibition of apoptosis on liver tumor development. 

Methods: We used male hepatocyte-specific knockout (KO) 

mice of Mcl-1, one of anti-apoptotic proteins, as a model of 

apoptosis-prone liver, and hepatocyte-specific single or double 

KO mice of Bak and Bax, proapoptotic proteins, as a model 

of apoptosis-resistant liver . To induce liver tumors those KO 

mice were intraperitoneally administered 20 mg/kg diethylnitrosamine 

(DEN) at the age of 2 weeks. Results: Mcl-1 KO 

mice spontaneous hepatocyte apoptosis as evidenced by HE 

staining of the liver sections, increasing serum ALT levels and 

serum caspase-3/7 activities. The number of TUNEL positive 

hepatocytes also increased in Mcl-1 KO mice. No liver tumors 

were observed at 6 months in both Mcl-1 KO and wild-type 

(WT) mice without DEN. On the other hand, while only 11.8% 

(2/17) of WT mice treated with DEN developed macroscopic 

liver tumors in 6 months, 100% (7/7) of Mcl-1 KO littermates 

developed (p


The following authors have nothing to disclose: Yasutoshi Nozaki, Satoshi 

Tanaka, Sadatsugu Sakane, Yugo Kai, Yuki Makino, Tasuku Nakabori, Yoshinobu 

Saito, Ryotaro Sakamori, Naoki Hiramatsu, Tomohide Tatsumi 

1986 

FGF15 and FGF19 Induce Disparate FGFR4-Mediated 

Hepatocarcinogenicity In-Vitro and In Two Murine Models: 

Implications for Drug-Associated Carcinogenicity 

Risk Assessments 

Lei Ling, Mei Zhou, Darrin Lindhout, Jian Luo, Michael Chen, 

Hong Yang, Mark Humphrey, Van Phung, Kalyani Mondal, Hugo 

Matern, Marc Learned, Stephen Rossi, Alex M. DePaoli, Hui Tian; 

NGM Biopharmaceuticals, S. San Francisco, CA 

Background and Aims: FGF19 is a regulator of bile acid (BA) 

synthesis in humans. However, FGF19 produces hepatocellular 

carcinoma (HCC) in transgenic mice and is linked to increased 

risk of HCC post-resection recurrence in humans. FGF15 is the 

rodent ortholog of human FGF19 with 50% amino acid (AA) 

homology and used in rodents to assess the pharmacologic and 

caricongenic activity of FGF19. Both FGF15 and FGF19 are 

potent inhibitors of Cyp7a1-mediated BA synthesis in rodents 

but the carcinogenicity risk of FGF15 is not well characterized. 

The hepatocarcinogenicity of FGF15 and FGF19 was 

evaluated in leptin receptor-deficient (db/db) mice after 24 

wks or diet-induced obese (DIO) mice after 52 weeks of treatment. 

In vitro receptor interaction and activation studies were 

performed. Methods: FGF15, FGF19 or control was dosed 

by long-term transgene expression using 1 dose of adeno-associated 

virus (AAV)-mediated gene delivery. Liver tissue was 

examined for tumors and liver weight at 24wks post-dose (db/ 

db) or 52wks post-dose (DIO). Gene expression of HCC-related 

and cell proliferation markers were measured in liver 

tissue. FGFR4-Klothoβ (FGFR4-KLB) receptor complex binding 

was assessed with SPR assays and transfected rat L6 cells were 

used to assess receptor activation. Results: FGF19 induced 

liver tumors in both models post-treatment at concentrations 

as low as 1 ng/ml (Figures 1a, 1b). In contrast, FGF15 failed 

to induce liver tumors at supraphysiologic concentrations and 

maintained normal liver weight and liver-body weight ratios. 

Liver tissue expression of Ki-67 (Figure 1c), α-fetoprotein, glypican-3, 

cyclin-a2, Ccnb1 and Ccnb2 were induced with FGF19 

but not FGF15. Reduced activation of FGFR4-KLB receptor complex 

was only seen with FGF15, as measured by decreased 

expression of STAT-3 target genes. These differences may be 

mediated by AA sequences unique to FGF15. Conclusions: 

Significant differences in hepatocellular proliferation and tumorigenesis 

were observed with FGF15 vs FGF19. These data 

demonstrate the potential challenges of assessing the risks of 

increased FGF19 levels in humans using FGF15 in rodent carcinogenicity 

models. 

Disclosures: 

Lei Ling - Employment: NGM Biopharmaceuticals, Inc. 

Mei Zhou - Stock Shareholder: NGM Biopharmaceuticals 

Darrin Lindhout - Patent Held/Filed: NGM Biopharmaceuticals; Stock Shareholder: 

NGM Biopharmaceuticals 

Jian Luo - Employment: NGM Biopharmaceuticals 

Hong Yang - Employment: NGM Biopharmaceuticals 

Mark Humphrey - Employment: NGM Biopharmaceuticals; Stock Shareholder: 

NGM Biopharmaceuticals 

Van Phung - Employment: NGM Biopharmaceuticals; Stock Shareholder: NGM 

Biopharmaceuticals 

Kalyani Mondal - Stock Shareholder: NGM BioPharmaceuticals 

Hugo Matern - Employment: NGM; Stock Shareholder: NGM 

Marc Learned - Employment: NGM Biopharmaceuticals, Inc.; Stock Shareholder: 

NGM Biopharmaceuticals, Inc. 

Stephen Rossi - Employment: NGM Biopharmaceuticals, Inc; Stock Shareholder: 

NGM Biopharmaceuticals, Gilead Sciences 

Alex M. DePaoli - Employment: NGM Biopharmaceuticals 

Hui Tian - Management Position: NGM Biopharma 

The following authors have nothing to disclose: Michael Chen 

1987 

Lymphotoxin–β regulated by NF–κB is a useful biomarker 

related to poor prognosis in hepatocellular carcinoma 

Takehisa Watanabe, Satomi Fujie, Youko Yoshimaru, Katsuya 

Nagaoka, Hiroko Setoyama, Kotaro Fukubayashi, Masakuni Tateyama, 

Hideaki Naoe, Jiro Fujimoto, Motohiko Tanaka, Yutaka 

Sasaki; Hepatology, Kumamoto Univ., Kunamoto, Japan 

The Lymphotoxin-β (LTβ) is a major pro-inflammatory cytokine 

that belongs to the TNF superfamily and mediates local 

inflammatory responses as a membrane-bound cytokine. The 

LTβ pathway is considered to be an important pathway in 

the hepato-carcinogenesis. We previously reported that the 

CCCTC-binding factor (CTCF) mediated higher order chromatin 

conformation of TNF/LT locus, including LTβ gene, is deeply 

involved in the regulation of LTβ expression in human HCC, 

by the global analysis with ChIP-chip and ChIP-seq. In this 

study, we attempted to reveal the molecular mechanism of LTβ 

regulation and the clinical characteristics of the patients with 

HCC expressing LTβ. Because chronic inflammation is one of 

the most crucial factors for hepato-carcinogenesis, we firstly 

focused on NF-κB, a major pro-inflammatory transcription factor, 

and investigated the influence of NF-κB on LTβ expression 

and higher order chromatin conformation, by using cell lines 

differed on the activity of NF-κB. As a result, LTβ was expressed 

continuously in the cells with constitutive active NF-κB in 

response to the stimulation with TNF. On the other hand, LTβ 

degraded rapidly in the cells without constitutive active NF-κB. 

These cells differ on the higher order chromatin conformation 

to control the interaction between the LTβ promoter and appropriate 

NF-κB responsive enhancer. These results indicated that 

NF-κB was involved in the higher order chromatin conformation 

and regulation of LTβ. Additionally, we found that LTβ 

could be quantified in the exosomes extracted from serum of 

the patient, whereas liver biopsy had been considered to be 

indispensable to describe how LTβ is expressed in hepatic lobules. 

To examine whether exosomal LTβ reflects LTβ expression 

in HCC tissue, we performed immunohistochemistry with LTβ 

antibodies in 24 samples from HCC patients and compare with 

the amount of exosomal LTβ. Consequently, high level exosomal 

LTβ reflected the high expression LTβ in the tumor tissues, 

not in the adjacent non-tumor tissues. Moreover, the patients 

with the high level exosomal LTβ exhibited significantly shorter 

survival time. Therefore, these results suggest that exosomal LTβ 

might be a useful biomarker related to poor prognosis in HCC. 

Disclosures: 

Yutaka Sasaki - Grant/Research Support: Chugai Pharmaceutical Co. JAPAN, 

MSD Co. JAPAN


The following authors have nothing to disclose: Takehisa Watanabe, Satomi 

Fujie, Youko Yoshimaru, Katsuya Nagaoka, Hiroko Setoyama, Kotaro Fukubayashi, 

Masakuni Tateyama, Hideaki Naoe, Jiro Fujimoto, Motohiko Tanaka 

1988 

DDB2 loss protects mice from H-ras12V-driven HCCs 

Dragana Kopanja, Akshay Pandey, Shuo Huang, Damiano Fantini, 

Pradip Raychaudhuri; Biochemistry and Molecular Genetics, 

University of Illinois at Chicago, Chicago, IL 

DDB2 (Damaged DNA-binding protein (DDB)-2,) is a protein 

encoded by the nucleotide excision repair (NER) gene xeroderma 

pigmentosum group E (XPE). DDB2 is involved in early 

steps of NER and in DNA damage-induced apoptosis. Interestingly, 

DDB2 also acts as transcriptional repressor and regulates 

expression of antioxidant genes MnSOD and Catalase. Furthermore, 

DDB2 represses transcription of EMT-inducing factors 

Snail, VEGF and ZEB1, and its loss in high grade colon cancers 

promotes metastasis by promoting EMT. Moreover, DDB2 

-/- mice develop spontaneous tumors at higher frequency than 

wt mice, suggesting that it can act as a tumor suppressor. In 

this study, we investigated role of DDB2 in liver cancer. Since 

Ras-signaling pathway is found to be ubiquitously activated 

in human hepatocellular carcinoma (HCC) through epigenetic 

silencing of the Ras-regulators, we used mice that express 

H-ras12V under control of albumin promoter. Male mice of 

this strain develop liver tumors at age of 8-9 months with penetrance 

of almost 100%. We crossed H-ras12V mice with DDB2 

knockout mice (DDB2 -/-). Surprisingly, in that model of HCCs, 

DDB2 deletion protects mice from liver tumor development. We 

observed lower number of liver tumor nodules in DDB2 knockout 

H-ras12V mice compared to the DDB2 +/+ H-ras12V mice. 

Moreover, unlike in colon cancer, DDB2 loss does not induce 

metastasis of H-ras12V driven liver cancer. To understand why 

DDB2 -/- H-ras12V mice develop lower number of tumor nodules 

compared to DDB2 +/+ H-ras12V, we harvested livers 

from 5 months old male mice of both genotypes. We found 

that DDB2 null livers have increased expression of MnSOD 

and decreased accumulation of ROS. Therefore, an impairment 

of oxidative damage could explain observed reduction in the 

number of H-ras12V-driven HCC nodules in DDB2 -/- mice. 

Since DDB2 is considered to act as a tumor suppressor, our 

finding that DDB2 loss protects mice from H-ras12V-driven liver 

tumor development is interesting and surprising, and it demonstrates 

that depending of the context of tumor development 

DDB2 can either inhibit or promote tumorigenesis. 

Disclosures: 

The following authors have nothing to disclose: Dragana Kopanja, Akshay Pandey, 

Shuo Huang, Damiano Fantini, Pradip Raychaudhuri 

1989 

Inflammation contributes to hepatocarcinogenesis 

through an acceleration of transcription-coupled mutagenesis 

Tomonori Matsumoto, Norihiro Nishijima, Tsutomu Chiba, Hiroyuki 

Marusawa; Department of Gastroenterology and Hepatology, 

Graduate School of Medicine, Kyoto University, Kyoto, Japan 

Chronic inflammation plays important roles in cancer development 

in association with various intrinsic mediators including 

cytokines and growth factors, however, accumulation of 

genetic alterations in tumor-related genes is a critical step 

required for malignant transformation. Activation-induced cytidine 

deaminase (AID), an intrinsic DNA mutator enzyme, has 

been demonstrated to be aberrantly elicited in epithelial cells 

by inflammatory stimulation, and to contribute to tumorigenesis 

by inducing genetic aberrations. To gain further insight into the 

inflammation-mediated genotoxic events required for carcinogenesis, 

we examined the role of chronic inflammation in the 

emergence of genetic aberrations in the liver with constitutive 

AID expression. Treatment with thioacetamide (TAA) at lowdose 

concentrations caused minimal hepatic inflammation in 

both wild-type and AID transgenic (Tg) mice. Surprisingly, all 

the TAA-treated AID Tg mice developed multiple liver cancers in 

6 months, while wild-type mice with TAA or AID Tg mice without 

TAA rarely develop tumors during the same periods. Whole 

exome sequencing demonstrated the enhanced accumulation 

of somatic mutations in various genes in the liver of TAA-treated 

AID Tg mice. Microarray analyses showed the transcriptional 

upregulation of several putative tumor suppressor genes under 

hepatic inflammatory conditions induced by TAA treatment. 

Additional quantitative reverse transcription-polymerase chain 

reaction analyses and deep-sequencing analyses revealed 

the transcriptional upregulation and enhanced mutagenesis 

of putative tumor suppressor genes, including dual specificity 

phosphatase 6 (Dusp6), early growth response 1 (Egr1) and 

inhibitor of DNA binding 2 (Id2), in AID-expressing liver with 

TAA-mediated hepatic inflammation. Together, these findings 

suggest that inflammation-mediated transcriptional upregulation 

of target genes, including putative tumor suppressor genes, 

enhances the opportunity for inflamed cells to acquire somatic 

mutations and contributes to the acceleration of tumorigenesis 

in the inflamed liver tissues. 

Disclosures: 

The following authors have nothing to disclose: Tomonori Matsumoto, Norihiro 

Nishijima, Tsutomu Chiba, Hiroyuki Marusawa 

1990 

Regulator of G-protein signaling 5 enhances portal vein 

invasion in hepatocellular carcinoma 

Yumi Umeno 1 , Sachiko Ogasawara 1 , Jun Akiba 1 , Hironori 

Kusano 1 , Osamu Nakashima 2 , Hironori Koga 3 , Takuji Torimura 3 , 

Hirohisa Yano 1 ; 1 Department of Pathology, Kurume University 

School of Medicine, Kurume, Japan; 2 Clinical Laboratory Medicine, 

Kurume University Hospital, Kurume, Japan; 3 Department of 

Medicine, Division of Gastroenterology, Kurume University School 

of Medicine, Kurume, Japan 

Aim: Hepatocellular carcinoma (HCC) is one of the most 

common cancers in the world and a leading cause of cancer 

death. Portal vein invasion (PVI) is one of the major prognostic 

factors in patients with HCC. The aim of this study is to identify 

molecules to regulate PVI. Materials and Methods: We 

selectively collected tissues from each part of cancerous area, 

paired noncancerous area and PVI area by using laser microdissection 

(LMD) and frozen sections obtained from 3 HCC 

patients. Subsequently, cDNA microarray was conducted. Five 

upregulated or downregulated molecules in plural cases were 

identified and subjected to the following examinations. In order 

to evaluate the expression and the relationship to clinicopathologic 

factors, quantitative RT-PCR (qRT-PCR) and immunohistochemical 

stain were performed in 32 HCCs and in 60 HCCs, 

respectively. Results: We focused on 3 upregulated molecules, 

i.e., integrin beta 3 (ITGB3), osteopontin (OPN) and regulator 

of G-protein signaling 5 (RGS5) and 2 downregulated molecules, 

i.e., metallothionein 1G (MT1G) and metallothionein 1H 

(MT1H) in PVI tissue compared with cancerous tissue by cDNA 

microarray analysis. In qRT-PCR, RGS5 was significantly overexpressed 

in cancerous tissue compared with noncancerous 

tissue (p = 0.02). However, there was no significant difference 

in ITGB3 and OPN expression (respectively: p = 0.13 and p 

= 0.12). The expression of MT1G and MT1H was significantly


lower in cancerous tissue compared with noncancerous tissue 

(respectively: p < 0.01 and p < 0.01). There were no significant 

differences between the expression of these five molecules 

and any clinicopathologic factors, including PVI. Immunohistochemical 

stain for RGS5 demonstrated that RGS5 expression 

was higher in cancerous tissue than in paired noncancerous 

tissue in 63.3% (38/60) of HCC. Furthermore, high expression 

of RGS5 in cancerous tissue was significantly associated with 

PVI (p < 0.01) and tended to be associated with intrahepatic 

metastasis (p = 0.0626). The cases with high expression of 

RGS5 in cancerous tissue were significantly frequent in simple 

nodular type with extranodular growth or confluent multinodular 

type than in simple nodular type (p < 0.01). Conclusion: By 

using LMD and cDNA microarray, we detected molecules associated 

with PVI. RGS5 expression in cancerous tissue was significantly 

upregulated at mRNA and protein levels compared 

with noncancerous tissue and was significantly associated with 

PVI of HCC. RGS5 may be a useful prognostic biomarker as 

well as a target of molecular therapy of HCC. In vitro functional 

analysis of RGS5 in HCC is underway. 

Disclosures: 

The following authors have nothing to disclose: Yumi Umeno, Sachiko Ogasawara, 

Jun Akiba, Hironori Kusano, Osamu Nakashima, Hironori Koga, Takuji 

Torimura, Hirohisa Yano 

1991 

Correlation between Incidence Rate of Hepatocellular 

Carcinoma and Chemical Air Pollution in the State of 

Texas. 

Ali Shirafkan 1 , Cristiana Rastellini 1 , Katherine Ensor 2 , Loren Raun 2 , 

Daria Zorzi 1 , Laura Campos 2 , Mauro Montalbano 1 , Tiziana Corsello 

1 , Luca Cicalese 1 ; 1 Department of Surgery, University of Texas 

Medical Branch, Galveston, TX; 2 Department of Statistics, Rice 

University, Houston, TX 

Introduction: Primary liver cancer is the third cause of cancer 

death in the world and the seventh in the United States. The 

known etiologies of Hepatocellular Carcinoma (HCC) are comprised 

of nonspecific cirrhosis, alcohol induced liver disease, 

hepatitis C (HCV) and B (HBV) infection. In addition, obesity 

and diabetes mellitus type two (T2DM) have been shown to 

increase the risk. Texas ranks third in the US with HCC incidence 

rate of 9.9 per 100,000. This is being compared to 

national average of 7.1 per 100,000, while distribution of the 

known risk factors in the state of Texas is similar to the national 

data. Texas has the largest oil and gas industry and is the 

second largest agricultural producer in the nation. Chemicals, 

fertilizers, herbicides and pesticides could potentially contribute 

to HCC development. Methods: The mean concentration 

value of Vinyl Chloride, Arsenic, Benzene and 1, 3 Butadiene 

reported by EPA (Environmental Protection Agency). Incidence 

rate of HCC in Texas by counties for the time period between 

2002 and 2010 was obtained from the Texas Cancer Registry. 

A scatter plot matrix, a logistic regression model and a generalized 

additive (GAM) logistic regression model were used 

to observe any correlation between these variables. Results: 

Increases in vinyl chloride and total Benzene and 1, 3 Butadiene 

were associated with an increased risk of HCC within a 

county. Arsenic levels appear protective at low levels, but at 

high levels the probability of HCC within a county increases. 

Conclusion: It is possible that vinyl chloride, arsenic, BZ and 1, 

3 BD produced in different areas of Texas probably contribute 

to the high incidence rates of HCC observed in this state. 

Disclosures: 

The following authors have nothing to disclose: Ali Shirafkan, Cristiana Rastellini, 

Katherine Ensor, Loren Raun, Daria Zorzi, Laura Campos, Mauro Montalbano, 

Tiziana Corsello, Luca Cicalese 

1992 

MicroRNA profiling of human cholangiocarcinoma 

Hong Joo Kim, Yong Kyun Cho, Woo Kyu Jeon, Byung Ik Kim; 

Internal Medicine, Sungkyunkwan University Kangbuk Samsung 

Hospital, Seoul, Korea (the Republic of) 

Background and aims: Cholangiocarcinoma (CC) is a highly 

malignant epithelial cancer of the biliary tract with high morbidity 

and mortality. In the current study, we performed microR- 

NAs (miRNAs) expression microarrays to identify the up- and 

downregulated miRNAs in CC. Thereafter, we performed oligomer 

and antagomir transfection experiments for the most prominently 

up- and downregulated miRNA in CC to identify the 

underlying molecular pathogenic mechanisms for proliferation, 

invasion and metastasis of CC. Methods: Human hepatic duct 

bifurcation site cancer cells SNU-1196 (KCLB No. 01196), 

and human common bile duct cancer cells SNU-245 (KCLB No. 

00245) were purchased from Korean Cell Line Bank and maintained 

in RPMI1640 medium. Affymetrix miRNA expression 

microarrays were used to determine the miRNA expression profile 

of each CC cell line and normal control cells obtained from 

primary culture of normal bile duct epithelium. Transfections 

were performed using Lipofectamine RNAiMAX (Invitrogen). 

Cell proliferation was determined by MTT assay after mimics 

or antisense oligonucleotide transfection. Western blotting of 

cell extracts was performed using rabbit monoclonal antibodies 

against human SRGAP2, Rac1 (both from BD Bioscience) and 

β-actin. Results: To profile the miRNA expression patterns in 

human CC cells, we compared the miRNA expression profile 

of 2 CC cell lines with that of normal control cells obtained 

from primary culture of normal bile duct epithelium. MicroR- 

NAs downregulated more than 10-folds log ratio in CC cell 

lines than normal control cells were as follows: miR-145-5p, 

miR-125b-5p, miR-125a-5p, and miR-100-5p. Upregulated 

miRNAs more than 8-folds log ratio in CC cell lines than normal 

control cells were as follows: miR-182-5p, miR-196a-5p, 

miR-194-5p, miR-192-5p, and miR-182-5p. Thereafter, we 

performed transfection experiments using the mimics and 

antagomirs targeting miR-145-5p. There were no significant 

differences in cell proliferation measured by MTT assay among 

the CC and normal control cell lines transfected with mimics 

and antagomirs. Downregulation of miR-145-5p by antagomir 

transfection was associated with increase in protein expression 

levels of SRGAP2 and Rac1. Conclusions: Our data identify 

that miR-145-5p is the most profoundly downregulated miRNA 

in human CC cells. SRGAP2 and Rac1 can be novel targets of 

miR-145-5p, establishing a role for miR-145-5p in modulating 

motility and metastasis of CC cells. Our data provide a possibility 

for further investigations for future miR-145-5p targeted 

approaches of anticancer treatment in.CC. 

Disclosures: 

The following authors have nothing to disclose: Hong Joo Kim, Yong Kyun Cho, 

Woo Kyu Jeon, Byung Ik Kim 

1993 

Cholangiocarcinoma cells interact with cancer-associated 

myofibroblasts in 3-D culture to provoke a strong 

desmoplastic-like reaction mediated by TGF-β. 

Alphonse E. Sirica, Akihiro Usui, Deanna J. Campbell; Pathology, 

Virginia Commonwealth University School of Medicine, Richmond, 

VA 

Intrahepatic cholangiocarcinomas (ICC) typically exhibit 

a prominent desmoplastic reaction marked by a dramatic 

increase in α-smooth muscle actin-positive cancer-associated 

fibroblasts (α-SMA+CAFs) together with the production of a


dense collagen-enriched fibrous stroma. In order to further our 

understanding of the biological relationships mediating the desmoplastic 

reaction in ICC, we employed a novel rat 3-D culture 

model to investigate fibrogenic promoting interactions between 

cholangiocarcinoma cells (BDEsp-TDE cc 

cells) and α-SMA+CAFs 

(BDEsp-TDF sm 

cells) when co-cultured in a dilute collagen-type I 

gel matrix. Both BDEsp-TDE cc 

and BDEsp-TDF sm 

cells were established 

from a desmoplastic rat cholangiocarcinoma from liver. 

Compared with monoculture controls, co-culturing BDEsp-TDE cc 

cholangiocarcinoma cells constitutively expressing TGF-β1 with 

BDEsp-TDF sm 

CAFs provoked an intense desmoplastic-like reaction 

within the gel culture matrix, characterized by a prominent 

deposition of dense collagen fibers, together with a significant 

increase in the numbers of BDEsp-TDF sm 

CAFs accumulated 

within the gel matrix. This reaction was greatly suppressed by 

the TGF-β receptor 1 kinase inhibitor LY2157299. Similarly, 

treatment of BDEsp-TDF sm 

cells cultured alone within 3-D gel cultures 

with TGF-β1 significantly increased the numbers of these 

SMA+ CAFs within the gel matrix (Figure), which correlated 

with an overproduction of dense collagen fibers, and which 

was also inhibited by >70% by LY2157299. These results 

strongly suggest that paracrine TGF-β signaling plays a major 

role in generating the desmoplastic reaction in ICC and that 

targeting this pathway may have important implications for 

stromal depletion therapy for desmoplastic ICC. Supported by 

NIH R01 CA 083650. 

TGF-β1-induced enhancement of BDEsp-TDF sm 

CAF cell accumulation 

within 3-D gel monoculture compared with vehicle control 

cultures (VC). 

associated with oxidation of lipids and formation of oxysterols. 

Oxysterols are allosteric regulators of Smoothened (Smo), the 

receptor of the Hedgehog (Hh) signaling pathway, which is 

activated in cholangiocarcinoma. The step wise process of 

malignant transformation of biliary epithelial cells is poorly 

described. Purpose: We aimed to interrogate malignant transformation 

of the biliary epithelium upon stimulation of the Hh 

pathway by oxysterols. Methods: The human non-malignant 

(H69 and NHC) and primary PSC patient-derived cholangiocyte 

cell lines were employed for this study. Upon treatment 

with oxysterols [7-keto-27-hydrocholesterol and 20(S)-hydroxycholesterol], 

cholangiocytes were analyzed for Hh pathway 

stimulation with Smo translocation to the plasma membrane 

and cilia by immunofluorescence, migration in modified Boyden 

chambers, and colony-formation assay in a soft-agar. A 

miRNA array was conducted to identify miRNAs that were 

induced in cholangiocytes in an oxysterol- and Smo-dependent 

manner. miRNA targets were analyzed in silico. Results: Cholangiocytes 

expressed Hh pathway components at a baseline 

(Gli2, Smo). Smo translocation to the plasma membrane and 

cilia were observed in the cells treated with oxysterols (10mM 

for 16 hours) but not in the vehicle treated cells. Stimulation 

with oxysterols (10 mM for 7 hours) lead to a 1.3 – 2 (p < 

0.01) fold increase in cholangiocyte migration. Oxysterols promoted 

anchorage-independent growth evidenced by formation 

of colony-like structures by non-malignant cholangiocytes in a 

soft-agar over the 6 week period. miR-1246 was upregulated 

(logFC = 2.7) in H69 cells upon stimulation with 7-keto-27-hydrocholesterol 

and was inhibited (logFC = -2.2) with Vismodegib, 

direct-inhibitor of Smo, pre-treatment. The Hh pathway 

inhibitory receptor Patched-1, Axin2, and GSK-3β, all of each 

are involved in Hh signaling and carcinogenesis, were found 

to be a target for miR-1246 in silico. Conclusions: Products of 

lipid oxidation, oxysterols, promote malignant transformation 

of cholangiocytes in the Hh pathway-dependent manner. The 

oxysterol-Smo axis upregulation of miR-1246 might provide a 

positive feedback mechanism in this transformation. 

Disclosures: 


The following authors have nothing to disclose: Nataliya Razumilava, Anuradha 

Krishnan, Steven P. O’Hara 

Disclosures: 

The following authors have nothing to disclose: Alphonse E. Sirica, Akihiro Usui, 

Deanna J. Campbell 

1994 

Oxysterols Promote Malignant Transformation of Cholangiocytes 

in a Hedgehog Signaling-dependent Manner 

Nataliya Razumilava 2 , Anuradha Krishnan 1 , Steven P. O’Hara 1 , 

Gregory J. Gores 1 ; 1 Mayo Clinic, Rochester, MN; 2 Division of 

Gastroenterology and Hepatology, University of Michigan, Ann 

Arbor, MI 

Background: Cholangiocarcinoma is a highly lethal neoplasm. 

It is often observed in a milieu of biliary tract inflammation 

1995 

A Randomized Prospective Open-label Trial Comparing 

Peginterferon Plus Adefovir or Tenofovir Combination 

Therapy Versus No Treatment in HBeAg-Negative 

Chronic Hepatitis B Patients with a Low Viral Load 

Annikki de Niet 1 , Louis Jansen 1 , Femke Stelma 1 , Sophie Willemse 1 , 

Sjoerd Kuiken 2 , Sebastiaan Weijer 3 , Carin M. Van Nieuwkerk 4 , 

Hans L. Zaaijer 5,1 , Richard Molenkamp 1 , Bart Takkenberg 1 , 

Maarten Koot 5 , Joanne Verheij 1 , Ulrich Beuers 1 , Hendrik W. 

Reesink 1 ; 1 Academic Medical Center, Amsterdam, Netherlands; 

2 Sint Lucas Andreas Hospital, Amsterdam, Netherlands; 3 Medical 

Center Zuiderzee, Lelystad, Netherlands; 4 VU Medical Center, 

Amsterdam, Netherlands; 5 Sanquin, Amsterdam, Netherlands 

Background and aims: Chronic hepatitis B (CHB) patients with 

a low viral load (LVL) are currently not eligible for antiviral 

treatment. However, they comprise the largest group of CHB 

patients and are still at increased risk of developing cirrhosis or 

hepatocellular carcinoma. Here, we present the final results of a 

randomized trial designed to determine HBsAg loss and decline 

in CHB patients with LVL receiving combination treatment of 

peginterferon alfa-2a (peg-IFN) and a nucleotide analogue or 

no treatment. Methods: 134 CHB patients (HBeAg-negative, 

HBV-DNA


peg-IFN + adefovir (arm I; n=46), peg-IFN + tenofovir (arm 

II; n=45) or no treatment (arm III; n=43) for 48 weeks (ITT 

population), and followed-up for an additional 24 weeks. Randomization 

was stratified by HBV genotype A (22%), non-A (B 

7%, C 4%, D 26%, E/F/G 20%), or indeterminable (21%). The 

median age was 43 years, 57% were male. Twelve patients 

discontinued the study before week 72 (5 in arm I, 6 in arm 

II, 1 in arm III). HBsAg loss (AxSYM


2008-2014 for >2 years (ETV group). We excluded patients 

who developed HCC within the first year of enrollment. The 

primary endpoint was the development of HCC, and secondary 

endpoints were the development of cirrhotic complications 

and mortality. Results As of Dec 31, 2014, we enrolled 1315 

patients in the ETV group and 367 untreated patients before the 

development of HCC. Compared to untreated group, patients in 

the ETV group were significantly older and had more advanced 

liver disease. The mean duration of follow-up was 7 and 4 

years in the untreated and ETV groups, respectively. Compared 

with the untreated group, ETV therapy was associated with a 

significantly lower lifetime risk for HCC (Logrank P


3.7 years and the median time from HBsAg loss to NA cessation 

was 0.6 years. At NA cessation, 61% of the patients 

were anti-HBs positive for a median duration of 0.5 years. 

After NA cessation, 4 (9%) patients relapsed. Three of them 

developed detectable HBV DNA (23, 223 and 1320 IU/mL) 

within the first 3 months. All 3 patients remained HBsAg negative, 

one was retreated while the other 2 returned to HBV 

DNA 1 

year and did not have significant co-morbidities were enrolled. 

Fasting serum, spot and 24 hour urine samples were collected. 

Fractional excretion of potassium (FEK + ), phosphate (FEPO 4 

), 

uric acid (FEUA), urinary β2MG/creatinine (Cr) and urine protein/creatinine 

ratio (UPCR) were calculated. The criteria of 

renal loss were defined as follow: proteinuria (24-hour urine 

protein >150 mg or positive protein dipstick), glucosuria while 

having normoglycemia, phosphate loss (FEPO 4 

>18%), uric 

acid loss (FEUA>15%), hypokalemia with renal K + loss, normal 

gap metabolic acidosis. Subclinical and definite proximal RTD 

was defined when there was a presence of 2 and ≥3 criteria, 

respectively. Receiver Operating Characteristic (ROC) was 

analyzed. The comparison of areas under the ROC was done. 

Results: Of 92 patients, subclinical and proximal RTD were 

seen in 15 (16.3%) and 9 (9.8%) patients. Median (range) 

duration of nucleotide analogue usage in normal, subclinical 

and definite proximal RTD were 45 (12-116), 66 (15-114), 

81 (42-108) months. The performance of spot urine protein, 

UPCR, urinary β2MG, urinary β2MG/Cr, FEPO 4 

and FEUA 

for detecting proximal RTD were showed in Table 1. Urinary 

β2MG yielded the highest AUROC [0.888 (0.804-0.972)]. At 

the cutoff of 300 mcg/L, urinary β2MG was 91.67% sensitivity 

and 76.47% specificity. There was no difference of diagnostic 

accuracy among tests (P=0.23). For detecting subclinical 

proximal RTD, urinary β2MG had the highest AUROC [0.822 

(0.699-0.946)]. At the cutoff of 300 mcg/L, urinary β2MG 

was 86.67% sensitivity and 77.27% specificity. Conclusions: 

16.3% and 9.8% CHB patients on nucleotide analogues developed 

subclinical and definite proximal RTD. Urinary β2MG 

had the highest accuracy for detecting proximal RTD. During 

nucleotide analogue treatment, urinary β2MG or UPCR should 

be monitored. 

Table 1. Receiver Operating Characteristic (ROC) analysis for 

detecting proximal RTD


Disclosures: 

Abhasnee Sobhonslidsuk - Grant/Research Support: Merck Sharp & Dohme 

corp, Eisai, Biotron; Speaking and Teaching: DKSH 

The following authors have nothing to disclose: Jirachaya Wanichanuwat, Bunyong 

Phakdeekitcharoen, Areepan Sophonsritsuk, Supanna Petraksa, Saowanee 

Kajanachumpol, Alongkorn Pugasab, Paisan Jittorntam, Anucha Kongsomgan, 

Pawin Numthavaj, Sittiruk Roytrakul 

2000 

Excellent outcomes of hepatitis B core antibody positive 

donors with oral antiviral agent alone in liver transplantation 

Tiffany C. Wong, James Fung, Kenneth S. Chok, See-Ching Chan, 

Chung Mau Lo; Surgery, The University of Hong Kong, Hong 

Kong, Hong Kong 

Background The use of hepatitis B core antibody (HBcAb) positive 

liver graft is controversial as antiviral protocol varies and 

the reported risk of de novo hepatitis B infection (HBV) ranged 

from 0 to 14%. Purpose To evaluate the risk of de novo HBV 

infection from HBcAb positive donor to Hepatitis B surface 

antigen (HBsAg) negative recipients in liver transplantation 

(LT). Method This was a retrospective review of all LT from 

2000 to 2014 in Queen Mary Hospital, Hong Kong. All data 

were retrieved from a prospective collected database. Results 

During study period, there were 1,034 LT at our center and 

128(12.4%) were from HBcAb positive donors to HBsAg negative 

recipients. Baseline demographics were listed in table 

1. All patients received antiviral prophylaxis only; lamivudine 

(n=117) and entecavir (n=11) with no hepatitis B immunoglobulin 

(HBIG) at any time point. De novo HBV as defined 

by HBsAg seropositive were seen in 3 (2.3%) recipients. They 

were 2 pediatric and 1 adult recipients. All had detectable 

HBV DNA (range from 179.3 to 373.6x10^6 copies/ml) at 

time of diagnosis and 2 had YMDD mutation. Liver biopsies 

were done in 2 of them and showed no evidence of fibrosis 

and immunostains for HBsAg and hepatitis B core antigen 

(HBcAg) werenegative. The median time from LT to de novo 

HBV was 59 (9-60) months. Tenofovir were added to the 2 

pediatric patients and adevofir was added to the adult patient. 

All patients with de novo HBV had undetectable HBV DNA after 

treatment. The median follow-up time was 71.1 (0.3 to 202.3) 

months. The 1-year and 5-year overall and graft survival were 

95.6%, 90.3% and 92.8% and 87.0% respectively. None had 

graft failure or mortality due to HBV infection. Conclusion The 

risk of de novo HBV infection is very low and had no impact on 

long-term patient and graft survival. Oral antiviral agent alone 

is effective and adequate to prevent de novo HBV in HBsAg 

negative patients who received a HBcAB positive graft. 

Table 1. Baseline demographic of our cohort 

Disclosures: 

The following authors have nothing to disclose: Tiffany C. Wong, James Fung, 

Kenneth S. Chok, See-Ching Chan, Chung Mau Lo 

2001 

Early prediction of response to peginterferon for 

HBeAg-positive chronic hepatitis B using HBsAg levels at 

week 4 and week 8 of therapy 

Milan J. Sonneveld 1 , Willem Pieter Brouwer 1 , Stefan Zeuzem 2 , Yilmaz 

Cakaloglu 3 , Fehmi Tabak 4 , Krzysztof. Simon 5 , Bettina E. Hansen 

1 , Harry L. Janssen 1,6 ; 1 Erasmus MC, University Medical Center 

Rotterdam, Rotterdam, Netherlands; 2 Medical Clinic 1, Johan 

Wolfgang Goethe University Medical Center, Frankfurt, Germany; 

3 Gastroenterohepatology, Istanbul Medical University, Istanbul, 

Turkey; 4 Hepatology, Cerrahpasa Medical School, Istanbul, Turkey; 

5 Infectious Disease, Medical University Wroclaw, Wroclaw, 

Poland; 6 Gastroenterology, University Health Network, Toronto, 

ON, Canada 

Background & Aims. Serum HBsAg levels can predict response 

to peginterferon (PEG-IFN) at week 12 of treatment; the performance 

of prediction-rules at earlier time-points is currently 

unknown. Methods. HBeAg-positive patients treated with PEG- 

IFN alfa-2b alone or in combination with lamivudine in a global 

randomized trial were enrolled if they had available outcome 

data and available HBsAg levels at week 4 and week 8 of 

therapy. Relationship between HBsAg at week 4 and week 8 

of treatment and response (HBeAg loss with HBV DNA


2002 

Final Results of Peginterferon Alfa-2b Add-on 

during Long-term Nucleos(t)ide Analogue Therapy in 

HBeAg-positive Patients – a Multicenter Randomized 

Controlled Trial (PEGON Study) 

Heng Chi 1 , Qing Xie 2 , Ning-Ping Zhang 3 , Xun Qi 4 , Chen Liang 4 , 

Simin Guo 2 , Qing Guo 2 , Pauline Arends 1 , Jiyao Wang 3 , Elke Verhey 

1 , Robert J. de Knegt 1 , Bettina E. Hansen 1 , Harry L. Janssen 1,5 ; 

1 Gastroenterology and Hepatology, Erasmus MC University Medical 

Center Rotterdam, Rotterdam, Netherlands; 2 Department of 

Infectious Diseases, Ruijin Hospital, Jiaotong University, Shanghai, 

China; 3 Department of Gastroenterology and Hepatology, Zhongshan 

hospital, Fudan University, Shanghai, China; 4 Department of 

Hepatitis Disease, Shanghai Public Health Clinical Center, Fudan 

University, Shanghai, China; 5 Toronto Centre for Liver Disease, 

Toronto General & Western Hospital, University of Toronto, Shanghai, 

ON, Canada 

BACKGROUND: Nucleos(t)ide analogues (NA) are potent 

inhibitors of viral replication in chronic hepatitis B (CHB) 

patients. However, sustained off-treatment response is infrequently 

achieved indicating the necessity of long-term therapy. 

Peginterferon (PEG-IFN) add-on in HBeAg-positive patients on 

long-term NA may facilitate serological responses and finite 

therapy. METHODS: In this investigator-initiated randomized 

trial conducted in Europe and China, 82 HBeAg-positive 

patients were treated for at least 12 months with entecavir 

(ETV) or tenofovir (TDF) with suppressed HBV DNA and HBeAg 

positivity at randomization. Patients were randomized to 48 

weeks of PEG-IFN alfa-2b add-on, or continued NA monotherapy. 

Response (HBeAg seroconversion with HBV DNA


improving PAGE-B scores developed a clinical event, whereas 

8/224 patients with stable and 3/71 patients with worsening 

PAGE-B scores did. Conclusion: In this large real-life European 

cohort of Caucasians and Asians treated with ETV, on-treatment 

PAGE-B score could predict HCC, as well as other liver-related 

and clinical events. Hence, the PAGE-B score obtained during 

ETV/TDF treatment could predict adverse clinical outcomes. 

Disclosures: 



Janssen, BMS 

Ashley S. Brown - Advisory Committees or Review Panels: MSD, Roche, Bristol-Myers-Squibb, 

Gilead, Janssen, Abbvie, Achillion; Speaking and Teaching: 

MSD, Roche, Bristol-Myers Squibb, Gilead, Janssen, Abbvie 


BMS 

David J. Mutimer - Advisory Committees or Review Panels: Gilead Sciences, 

AbbVie, Janssen, MSD, BMS; Speaking and Teaching: Gilead Sciences, AbbVie, 

Janssen, BMS 




Inc., Abbvie 





























The following authors have nothing to disclose: Heng Chi, Willem Pieter Brouwer, 

Massimo Fasano, Katja Deterding, Ye H. Oo, Pauline Arends, Teresa A. Santantonio, 

Pierre Pradat, Bettina E. Hansen 

2004 

Long Term Efficacy and Safety of Tenofovir DF (TDF) in 

Chronic Hepatitis B Patients (CHB) with Documented 

Lamivudine Resistance (LAM-R): 5 Year Results From a 

Randomized, Controlled Trial 

Scott Fung 1 , Hie-Won Hann 2 , Magdy Elkhashab 3 , Thomas Berg 4 , 

Milotka J. Fabri 5 , Andrzej Horban 6 , Mijomir Pelemis 7 , Ioan 

Sporea 8 , John F. Flaherty 9 , Benedetta Massetto 9 , Kyungpil Kim 9 , 

Kathryn M. Kitrinos 9 , Mani Subramanian 9 , Cihan Yurdaydin 10 , 

Edward J. Gane 11 ; 1 Toronto General Hospital, Toronto, ON, 

Canada; 2 Thomas Jefferson University Hospital, Philadelphia, PA; 

3 Toronto Liver Centre, Toronto, ON, Canada; 4 University Hospital 

Leipzig, Leipzig, Germany; 5 Clinic for Infectious Diseases, 

Novi Sad, Serbia; 6 Medical University of Warsaw, Warsaw, 

Poland; 7 Clinic for Infectious and Tropical Diseases,, Belgrade, 

Serbia; 8 University of Medicine and Pharmacy Timisoara, Timisoara, 

Romania; 9 Gilead Sciences, Inc., Foster City, CA; 10 Ankara 

Üniversitesi, Ankara, Turkey; 11 Auckland General Hospital, Auckland, 

New Zealand 

Background: In CHB patients with LAM-resistance (LAM-R), TDF 

has shown efficacy comparable to FTC/TDF and no detectable 

TDF resistance at 2 years (Gastroenterology 2014;146:980- 

88). The final 5 year efficacy and safety results from this trial 

are presented. Methods: CHB patients on LAM with HBV DNA 

≥3 log 10 

IU/mL and with documented LAM-R (INNO-LiPA 

Multi-DR, v3) were randomized (1:1) to TDF or FTC/TDF and 

followed in a blinded fashion for 5 years. Results: Two hundred 

eighty patients were randomized; 239 (85%) completed 5 

years of treatment. At baseline, mean age was 47 years, most 

were male (75%) and non-Asian (66%); 53% were HBeAgnegative, 

and HBV genotype distribution (A-D) was 22%, 13%, 

19%, and 43%, respectively. Mean (SD) HBV DNA was 5.7 

(1.9) log 10 

IU/mL, and 42% had ALT ≤ULN at baseline. At 

Year 5, virologic, serologic, and biochemical responses were 

similar among groups, and remained stable from Year 2 to 5 

(Table). Nine patients (4-TDF, 5-FTC/TDF) discontinued due to 

an adverse event, including increased serum creatinine in 1 

patient. Hepatocellular carcinoma was reported in 4 (1.4%) 

patients. Confirmed renal safety endpoints (both groups combined) 

over 5 years were: CrCL


Disclosures: 



Hie-Won Hann - Advisory Committees or Review Panels: Gilead; Grant/Research 

Support: Gilead, Bristol-Myers Squibb 

Magdy Elkhashab - Advisory Committees or Review Panels: GSK, Gilead, Merk, 

BMS, ABBVIE; Grant/Research Support: EISAI, GENENTECH, GSK, GILEAD, 

ABBVIE, Merk, BMS 







John F. Flaherty - Employment: Gilead Sciences; Stock Shareholder: Gilead Sciences 




Sciences 


Cihan Yurdaydin - Advisory Committees or Review Panels: Janssen, Roche, 

Merck, Gilead, AbbVie; Speaking and Teaching: BMS 




The following authors have nothing to disclose: Milotka J. Fabri, Andrzej Horban, 

Mijomir Pelemis, Ioan Sporea, Kyungpil Kim 

2005 

Plasma MicroRNA Levels are Associated with Therapy 

Response in Chronic Hepatitis B Patients Treated with 

Peginterferon and Adefovir 

Meike H. van der Ree 1,2 , Louis Jansen 1,2 , Karel A. van Dort 2 , Bart 

Takkenberg 1 , Neeltje A. Kootstra 2 , Hendrik W. Reesink 1,2 ; 1 Gastroenterology 

and Hepatology, AMC, Amsterdam, Netherlands; 

2 Experimental Immunology, AMC, Amsterdam, Netherlands 

Background and Aims: MicroRNAs (miRNAs) are small, 

non-coding RNA molecules involved in various cellular processes 

of post-transcriptional regulation of gene expression. 

Circulating miRNAs have been linked to hepatitis B virus (HBV) 

infection. The aim of this study was to identify plasma miRNAs 

signatures at baseline that predict response to peginterferon 

based therapy in chronic hepatitis B (CHB) patients. Methods: 

We included 86 CHB patients who participated in an investigator-initiated 

study and completed 48 weeks of peginterferon 

alfa-2a and adefovir combination therapy, followed by 

a treatment-free follow-up of 2 years (week 144). RNA was 

isolated from baseline plasma samples with the miRCURY RNA 

isolation kit (Exiqon). A miRNA profile (qPCR panel of 179 

miRNAs, Exiqon) was determined in an identification cohort 

of 12 HBeAg-positive and 12 HBeAg-negative patients, both 

consisting of 6 non-responders (NR) and 6 combined responders 

(CR) (HBeAg negativity, HBV DNA ≤2,000 IU/mL, and ALT 

normalization) of which 3 patients had HBsAg loss. Then, a 

selection of 28 miRNAs (>1.5-fold difference (p2-fold difference (p


ings may be significant enough to even warrant a Black Box 

Warning by the FDA 

Disclosures: 

John R. Lake - Advisory Committees or Review Panels: BMS; Consulting: Vital 

Therapies, Novartis; Grant/Research Support: Gilead, Salix, Ocera, Essai 

The following authors have nothing to disclose: Amar Mahgoub, Tyson Sievers 

2007 

TKM-HBV, a Novel RNA Interference Treatment for 

Chronic Hepatitis B, Rapidly Reduces Surface Antigen 

and other Viral Proteins in both Intrahepatic and Peripheral 

Compartments 

Amy C. Lee, Emily P. Thi, Luying Pei, Xin Ye, Jennifer Cross, 

Sandie Y. Du, Ammen P. Dhillon, Janet R. Phelps, Kevin McClintock, 

Michael Abrams, Ian MacLachlan; Tekmira Pharmaceuticals, 

Burnaby, BC, Canada 

TKM-HBV is a novel RNA interference (RNAi) therapeutic intervention 

against hepatitis B virus (HBV) and currently in Phase 1 

clinical development. It is designed to reduce the viral antigen 

load in chronically infected patients and allow the body to 

escape the state of immune repression imposed by the virus. 

A mixture of three different oligonucleotide duplexes encapsulated 

within a lipid nanoparticle delivery system, TKM-HBV acts 

directly on all HBV RNAs (pregenomic RNA as well as viral 

mRNA) via nucleotide sequence-specific cleavage. Because it 

prevents the synthesis of viral proteins and reduces the overall 

antigen load in the body, this mode of drug action may 

present advantages over other approaches that seek to block 

viral protein secretion into the bloodstream thereby potentially 

causing intracellular build-up and ER stress. A hydrodynamic 

injection (HDI) immunodeficient NOD.CB17-Prkdc scid /J mouse 

model of HBV was used to characterize viral RNA targeting 

and the time course of viral antigen reduction following a single 

administration of TKM-HBV and its effective distribution 

to its target organ, the liver. HBV RNA was measured using 

branched DNA assays, HBV DNA was quantitated via QPCR, 

and HBV proteins were detected using ELISA and immunohistochemistry 

methods. Over 92% reduction of total HBV RNA in 

the liver was observed as soon as 2 days after treatment, with 

a slightly more moderate effect on the 3.5 kb species alone 

suggesting this molecule is less accessible to the cellular RNAi 

machinery than other HBV transcripts. Onset of serum HBV 

DNA reduction was also highly rapid, with the nadir of >98% 

inhibition detected at Day 2. Maximal treatment effects were 

essentially reached at Day 4 for intrahepatic and serum HBsAg 

(98% and 97%, respectively) whereas reduction of intrahepatic 

HBcAg appeared to be a more gradual process, continuing to 

decrease beyond Day 4 with a nadir reached at Day 7. Taken 

together, these data suggest that the peripheral compartment is 

a dynamic environment with rapid turnover of viral markers in 

the blood stream whereas the turnover of viral proteins within 

the intracellular compartment of liver cells occurs more slowly. 

The kinetics of viral marker turnover may differ in a more 

immunocompetent system. In summary, TKM-HBV effectively 

removed viral antigens from both the intrahepatic and peripheral 

compartments within days after a single treatment dose 

in HDI mice. These viral elements include immunomodulatory 

surface and core proteins which are implicated in mediating 

the immune-repressed condition of chronic HBV infection. 

Disclosures: 

Amy C. Lee - Employment: Tekmira 

Emily P. Thi - Employment: Tekmira Pharmaceuticals 

Xin Ye - Employment: Tekmira Pharmaceuticals Corporation 

Kevin McClintock - Employment: Tekmira Pharmaceuticals, Tekmira Pharmaceuticals, 

Tekmira Pharmaceuticals, Tekmira Pharmaceuticals 

The following authors have nothing to disclose: Luying Pei, Jennifer Cross, Sandie 

Y. Du, Ammen P. Dhillon, Janet R. Phelps, Michael Abrams, Ian MacLachlan 

2008 

Long-Term Clinical And Serological Outcome Of Hepatitis 

B E-Antigen Negative Chinese-Pegylated Interferon 

Versus Prolonged Tenofovir Disoproxil Fumarate Or 

Entecavir-Eight Years Follow-Up 

Guofeng Chen 1 , Qing Shao 1 , Dong Ji 1 , Fan Li 1 , Bing Li 1 , Zhongbin 

Li 1 , Jialiang Liu 1 , Xiaoxia Niu 1 , Yudong Wang 2 , Cheng Wang 2 , 

Jing Chen 2 , Vanessa Wu 2 , George K. Lau 1,2 ; 1 Liver Fibrosis Diagnosis 

and Treatment Center, 302 Hospital, Beijing, China; 2 Gastroenterology 

& Hepatology, Humanity & Health Medical Group, 

Hong Kong, China 

Background and Aims Long-term follow-up clinical and serological 

data after a finite course of 48 wks pegylated interferon-based 

therapy as compared with prolonged nucleos(t) 

ide analogues (NUCs) therapy in HBeAg negative Chinese 

patients, is lacking. Methods In a single center, 121 HBeAg 

negative Chinese patients treated with a 48 wks course of 

pegylated interferon (pIFN) were compared with 346 nucleos(t)ide-naïve 

patients treated with prolonged TDF (n=158) or 

ETV (n=188) for over eight years. Baseline liver fibrosis was 

assessed by either liver biopsy or by Fibroscan (Echosens). 

All patients were followed up at 3-6 monthly intervals, with 

serial measurement of liver function test, serum HBV DNA, 

HBsAg, anti-HBs (if HBsAg turn negative) and liver stiffness 

by fibroscan. Quantitative HBsAg (qHBsAg), with the use of 

either Architect HBsAg (Abbott Laboratories, Ireland) or Elecsys 

HBsAg (Roche Diagnostics, Germany) was also performed on 

all serum samples available. Multivariable regression analysis 

was performed to identify the independent predictors of HBsAg 

loss, liver fibrosis regression and development of hepatocellular 

carcinoma. Results Twenty-four (19.8%) pIFN-treated as 

compared to 9 (2.6%) NUCs-treated patients had serological 

clearance of HBsAg (p


2009 

Functional Activation of NK and CD8 + T Cells In Vitro by 

the Toll-Like Receptor 7 Agonist GS-9620 

Holly Micolochick Steuer 1 , Stephane Daffis 1 , Sophie M. Lehar 1 , 

Adam Palazzo 1 , Hugo Tharinger 1 , Christian Frey 1 , Stefan Pflanz 1 , 

Congrong Niu 1 , Christine Y. Chang 2 , Michelle Q. Jin 2 , Vincent L. 

Chen 3 , William E. Delaney 1 , Leanne Peiser 1 , Simon P. Fletcher 1 , 

Mindie H. Nguyen 2 ; 1 Gilead Sciences, Foster City, CA; 2 Division 

of Gastroenterology and Hepatology, Stanford University Medical 

Center, Palo Alto, CA; 3 Department of Medicine, Stanford University 

Medical Center, Palo Alto, CA 

Background & Aims: GS-9620, an oral agonist of toll-like receptor 

7 (TLR7), is in phase 2 studies for the treatment of chronic 

hepatitis B (CHB). We previously demonstrated that reduction 

of HBsAg levels by GS-9620 in HBV-infected chimpanzees was 

associated with the intrahepatic induction of NK and CD8 + T 

cell transcriptional signatures, together with biochemical and 

immunohistochemical evidence of hepatocyte cell death. To 

further explore the cellular mechanisms of antiviral response to 

GS-9620, we characterized the effect of GS-9620 on primary 

human NK and CD8 + T cells in vitro. Methods: Peripheral blood 

mononuclear cells (PBMC) obtained from healthy volunteers 

(HV) and CHB patients (HBeAg-positive and HBeAg-negative 

patients, treatment naïve and virally suppressed with nucleos(t) 

ide therapy) were treated with 1-100 nM GS-9620. NK cell 

activation and effector function in HV PBMCs was analyzed 

by flow cytometry, and cytolytic activity was measured with a 

fluorescent cell-based cytotoxicity assay. CD8 + T cell function 

was assessed at recall by flow cytometry after expansion of HV 

or CHB PBMCs for 6-9 days with CEFT (CMV, EBV, Influenza 

and Tetanus toxin) or HBV core peptides, respectively. Results: 

GS-9620 potently induced expression of the activation marker 

CD69 on NK cells from HV PBMCs, and augmented NK cell 

cytotoxicity against HepG2 cells and HBV-infected primary 

human hepatocytes (PHH). In contrast, GS-9620 did not induce 

intracellular IFN-γ production in NK cells. GS-9620 induced 

comparable IFN-α levels in HV (n=5) and CHB PBMCs (n=11) 

(mean 1403 vs. 1075 pg/mL, respectively, p=0.44). This was 

notable because receptor blocking experiments demonstrated 

that GS-9620-dependent NK cell activation was mediated 

through type I IFN. GS-9620 also increased the frequency of 

CD8 + T cells expressing the cytotoxic marker CD107a after 

HBV core peptide expansion of CHB PBMCs (n=11, mean 

1.9-fold increase, p=0.03). Similarly, GS-9620 increased the 

frequency of CD107a + CD8 + T cells in HV PBMCs after CEFT 

peptide expansion (n=9, mean 2.1-fold increase, p=0.03). 

In contrast, GS-9620 did not alter the frequency of IFN-γ or 

TNF-α secreting CD8 + T cells in either HV or CHB PBMCs after 

peptide expansion. Conclusion: GS-9620 induced a cytotoxic 

phenotype but not antiviral cytokine production in human NK 

and CD8 + T cells in vitro. These in vitro functional data suggest 

that the antiviral response to GS-9620 in vivo is likely mediated, 

at least in part, by the cytolytic activity of NK and/or 

CD8 + T cells. 

Disclosures: 

Holly Micolochick Steuer - Employment: Gilead Sciences, Pharmasset, Inc, Pharmasset, 

Inc, Pharmasset, Inc 

Stephane Daffis - Employment: Gilead Sciences 

Adam Palazzo - Employment: Gilead Sciences; Stock Shareholder: Gilead Sciences 

Christian Frey - Employment: Gilead Sciences, Inc.; Stock Shareholder: Gilead 


Stefan Pflanz - Employment: Gilead Sciences 

Congrong Niu - Employment: Gilead Science 

William E. Delaney - Employment: Gilead Sciences; Patent Held/Filed: Gilead 

Sciences; Stock Shareholder: Gilead Sciences 

Leanne Peiser - Employment: Gilead Sciences, Merck; Stock Shareholder: Gilead 

Sciences, Merck 

Simon P. Fletcher - Employment: Gilead Sciences; Stock Shareholder: Gilead 

Sciences 





The following authors have nothing to disclose: Sophie M. Lehar, Hugo Tharinger, 

Christine Y. Chang, Michelle Q. Jin, Vincent L. Chen 

2010 

The incidence and predictors of HBsAg loss and hepatocellular 

carcinoma development after the cessation of 

lamivudine and entecavir treatment in chronic hepatitis 

B patients 

Chien-Hung Chen, Chuan-Mo Lee, Chao-Hung Hung, JIng-Houng 

Wang, Sheng-Nan Lu, Tsung-Hui Hu; Chang Gung Memorial Hospital-Kaohsiung 

Medical Center, Kaohsiung, Taiwan 

Background: It remained unclear that the incidence and predictors 

of hepatitis B surface antigen (HBsAg) loss and hepatocellular 

carcinoma (HCC) development after the cessation of 

nucleos(t)ide analogs (NA) therapy. Aims: To investigate the 

incidence and predictors of HBsAg loss and HCC developement 

after stopping NA treatment in chronic hepatitis B (CHB) 

patients. Patients and Methods: From 2004 to 2012, a total 

of 450 CHB patients (169 HBeAg-positive, 281 HBeAg-negative) 

(84 cirrhosis) received lamivudine (n=198) or entecavir 

(n=252) treatment and have stopped the treatment at least 12 

months were recruited. All patients fulfilled the stopping criteria 

of the APASL 2008 or 2012. Results: In 169 HBeAg-positive 

patients, the cumulative rates of HBsAg loss in years 3, 5 and 

8 were 3.6%, 9.9% and 18% respectively, after stopping NA 

treatment. Cox regression analysis showed that old age, lower 

baseline HBV DNA and end-of-treatment qHBsAg were independent 

predictors for HBsAg loss. In 281 HBeAg-negative 

patients, the cumulative rates of HBsAg loss in years 3, 5 and 

8 were 14.3%, 26.8% and 45.4% respectively, after stopping 

NA treatment. Cox regression analysis showed that male, cirrhosis, 

lower baseline ALT and end-of-treatment qHBsAg levels 

were independent predictors for HBsAg loss. Of the 56 patients 

with HBsAg loss, 8 and 4 experienced virological and clinical 

relapse respectively, before HBsAg loss. For all patients, the 

cumulative rates of HBsAg loss in patients who had end-oftreatment 

qHBsAg 1000 IU/mL 

in year 7 were 71.9%, 45.1%, 27.3% and 7.5% respectively. 

For all patients, the cumulative rates of new HCC development 

in year 10 (follow-up period included re-treatment duration) in 

non-cirrhotic and cirrhotic patients were 2% and 33% respectively. 

Cox regression analysis showed that old age, cirrhosis 

and longer treatment duration were independent predictors for 

new HCC development. HBsAg loss after stopping NA treatment 

was not a significant factor for HCC development. Five 

patients (4 cirrhosis) who experienced HBsAg loss developed 

new HCC. Conclusions: End-of-treatment qHBsAg level was a 

useful predictor for HBsAg loss after stopping lamivudine and 

entecavir treatment in CHB patient. HBsAg loss after stopping 

NA treatment did not completely eliminate the risk of HCC, 

especially in cirrhotic patient. 

Disclosures: 

The following authors have nothing to disclose: Chien-Hung Chen, Chuan-Mo 

Lee, Chao-Hung Hung, JIng-Houng Wang, Sheng-Nan Lu, Tsung-Hui Hu


2011 

The combination of IFN-Lambda-4 and HLA-DPB1 polymorphisms 

accurately predict HBsAg loss in interferon 

treated genotype D patients with HBeAg-negative 

chronic hepatitis B 

Pietro Lampertico 1 , Enrico Galmozzi 1 , Floriana Facchetti 1 , Cristina 

Cheroni 2 , Federica Invernizzi 1 , Vincenza Valveri 2 , Roberta 

Soffredini 1 , Mauro Viganò 3 , Sergio Abrignani 2 , Raffaele De Francesco 

2 , Massimo Colombo 1 ; 1 Division of Gastroenterology and 

Hepatology, Fondazione IRCCS Ca’ granda Ospedale Maggiore 

policlinico, Università degli Studi di Milano, Milan, Italy; 2 INGM 

- Istituto Nazionale Genetica Molecolare “Romeo ed Enrica Invernizzi”, 

Milan, Italy; 3 Division of Hepatology, Ospedale San 

Giuseppe, Università degli Studi di Milano, Milan, Italy 

Background. HLA-DP polymorphisms have recently been associated 

to spontaneous hepatitis B virus (HBV) clearance in 

Asians patients whereas polymorphisms mapping the interferon 

lambda 3 and 4 (IFNL3 and 4) genes predicted the outcome 

of IFN-based therapy of HCV infection. Whether the latter 

applies also to HBsAg clearance in IFN treated difficult to cure 

HBeAg-negative genotype D chronic hepatitis B Caucasian 

patients, is unknown. Methods. 126 such patients with compensated 

disease (46 years, 82% males, 90% genotype D, 

HBV DNA 6.2 log cp/ml, ALT 132 IU/L, 40% with cirrhosis) 

received (Peg)IFN alfa for 22 (6-48) months and were followed 

for 11 (1-23) years with HBsAg clearance as a primary endpoint. 

HLA-DPA1 (rs3077 A/G), HLA-DPB1 (rs9277534 A/G 

and rs9277535 A/G), IFNL3 (rs8099917 T/G) and IFNL4 

(rs12979860 C/T, rs368234815 TT/ΔG and rs117648444 

C/T) polymorphisms were assessed by both TaqMan SNP 

Genotyping Assays and Sanger sequencing. Results. Twenty-eight 

patients ultimately cleared HBsAg with a 18-year 

cumulative probability of 30%, with a preference for patients 

with lower HBV DNA levels and higher ALT levels at baseline, 

HLA-DPB1 AG/GG, HLA-DPA1 AG/GG, IFNL3 rs8099917 

TT genotypes and the combination of IFNL4 rs368234815 

TT/TT and rs117648444 CT/TT genotypes. At multivariate 

analysis, HLA-DPB1 rs9277535 AG/GG genotype was the 

strongest independent baseline predictor of HBsAg seroclearance 

(HR: 6.61; 95%CI 2.6-16, p5 yrs 

(mean follow-up:6.4, median:6.2). Results: HCCs have been 

diagnosed in 90/1946 (4.6%) patients within the first 5 yrs 

and 7/794 (0.9%) patients remaining at risk beyond the first 5 

yrs of therapy. The 7 HCCs diagnosed after yr-5 (at 5.4-6.7 yrs 

after ETV/TDF onset) developed in 4/531 (0.8%) non-cirrhotic 

and 3/224 (1.3%) cirrhotic male patients [50-75 yrs old at 

ETV (n=2) or TDF (n=5) onset] who had remained in on-therapy 

virological remission since the first yr (the 75-yr old patient had 

discontinued TDF at 5 yrs and had normal ALT and HBVDNA 


yrs:1.07%, p=0.046). HCC development beyond yr-5 was 

associated with older age (RH per yr:1.069, 95% CI:0.997- 

1.146; p=0.062) or age ≥55 yrs at ETV/TDF start (p=0.029 

by log-rank), but not with gender (p=0.340) or presence of 

cirrhosis at baseline (p=0.371). Conclusions: The HCC risk 

seems to be decreasing after the first 5 yrs of ETV/TDF therapy 

in CHB patients, especially in those with compensated cirrhosis 

at baseline. Older age at treatment initiation, particularly age 

≥55 yrs, appears to represent the main risk factor associated 

with late HCC development. 

Disclosures: 








Janssen, BMS 




Inc., Abbvie 

Ioannis Goulis - Consulting: Gilead Sciences, Abbvie, Janssen-Cilag, Janssen-Cilag, 

BMS; Grant/Research Support: BMS; Speaking and Teaching: BMS, Gilead 

Sciences, Janssen-Cilag 

Spilios Manolakopoulos - Advisory Committees or Review Panels: NOVARTIS, 

ABBVIE, MSD, BMS, GILEAD, JANNSEN; Consulting: GILEAD, BMS, ABBVIE, 

JANNSEN, MSD; Grant/Research Support: BMS, GILEAD; Speaking and Teaching: 

ABBVIE, MSD, GILEAD, BMS, GSK, JANNSEN 

















Rafael Esteban - Speaking and Teaching: MSD, BMS, Novartis, Gilead, Glaxo, 

MSD, BMS, Novartis, Gilead, Glaxo, Janssen 






Pietro Lampertico - Advisory Committees or Review Panels: Bayer, Bayer; Speaking 

and Teaching: Bristol-Myers Squibb, Roche, GlaxoSmithKline, Novartis, Gilead, 

Bristol-Myers Squibb, Roche, GlaxoSmithKline, Novartis, Gilead 

The following authors have nothing to disclose: Ramazan Idilman, George N. 

Dalekos, Heng Chi, José Luís Calleja, Vana Sypsa, Federica Invernizzi, Spyros I. 

Siakavellas, Onur Keskin, Nikolaos Gatselis, Bettina E. Hansen, Maria Lehretz, 

Juan de la Revilla, Savvoula Savvidou, Anastasia Kourikou, Floriana Facchetti, 

Jiannis Vlachogiannakos, Kostas Galanis 

2013 

Week 24 HBsAg levels predict response to peginterferon 

alfa-2b in HBeAg-positive genotype B or C 

chronic hepatitis B patients not meeting a week 12 stopping-rule 

Milan J. Sonneveld 1 , Jun Cheng 2 , Qing Xie 3 , Bettina E. Hansen 1 , 

Jin-Lin Hou 4 , Fehmi Tabak 5 , Stefan Zeuzem 6 , Harry L. Janssen 1,7 ; 

1 Erasmus MC, University Medical Center Rotterdam, Rotterdam, 

Netherlands; 2 Capital Medical University, Ditan Hospital, Beijing, 

China; 3 Infectious Diseases, Ruijin Hospital, Shanghai, China; 

4 Nanfang Hospital, Infectious Diseases, Guangzhou, China; 

5 Istanbul Medical School, Hepatology, Istanbul, Turkey; 6 Medical 

clinic 1, Goethe University, Frankfurt, Germany; 7 Gastroenterology, 

University Health Network, Toronto, ON, Canada 

Background & Aims. Serum levels of HBsAg (qHBsAg) predict 

response in HBeAg-positive patients with HBV genotypes B 

and C treated with peginterferon (PEG-IFN). A stopping-rule 

has been proposed based on qHBsAg >20,000 at week 

12 or 24. The performance of these stopping-rules has not 

been sufficiently validated in patients treated with PEG-IFN 

alfa-2b. Similarly, the value of measuring qHBsAg at week 24 

in patients not meeting the week 12 stopping-rule is unknown. 

Methods. HBeAg-positive patients with HBV genotypes B or 

C treated with PEG-IFN alfa-2b monotherapy for one year in 

2 global randomized trials (HBV 99-01 and P05170) were 

enrolled if they had available qHBsAg and a known therapy 

outcome. We assessed (1) the performance of the proposed 

stopping-rules based on qHBsAg >20,000 IU/mL and (2) 

assessed predictive performance of qHBsAg at week 24 in 

patients not meeting the week 12 stopping-rule. Response was 

defined as HBeAg loss with HBV DNA 20,000 at week 

12 was observed in 53 (26%) patients, of whom 4 (7.5%) 

achieved a response (negative predictive value [NPV] 92%). 

At week 24, 33 (16%) patients had qHBsAg >20,000, of 

whom 0 (0%, NPV 100%) achieved a response (p20,000 

IU/mL) at week 12 or 24 of PEG-IFN alfa-2b therapy identifies 

genotype B and C patients with a low probability of response. 

Among patients not meeting the week 12 stopping-rule, qHBsAg 

at week 24 predicts treatment outcome. 

Disclosures: 

Milan J. Sonneveld - Advisory Committees or Review Panels: Roche; Speaking 

and Teaching: Roche, BMS 


& Co., Janssen







The following authors have nothing to disclose: Jun Cheng, Qing Xie, Bettina E. 

Hansen, Jin-Lin Hou, Fehmi Tabak 

not achieve significant HBsAg declines early in treatment and 

highlights the need for new therapies to more potently inhibit 

cccDNA transcription or persistence. 

2014 

Early Decline in Serum HBsAg at Week 4 and 12 

of Therapy with Tenofovir Disoproxil Fumarate and 

Pegylated Interferon is Predictive of HBsAg Loss 

Henry Lik-Yuen Chan 1 , Sang Hoon Ahn 2 , Wan-Long Chuang 3 , 

Fehmi Tabak 4 , Rajiv Mehta 5 , Joerg Petersen 6 , George Wu 7 , Eduardo 

B. Martins 7 , Leland J. Yee 7 , Anuj Gaggar 7 , Mani Subramanian 

7 , Seng Gee Lim 8 , Scott Fung 9 , Graham R. Foster 10 , Maria 

Buti 11 , Giovanni B. Gaeta 12 , Aric Josun Hui 13 , George V. Papatheodoridis 

14 , Robert Flisiak 15 , Patrick Marcellin 16 ; 1 The Chinese 

University of Hong Kong, Hong Kong, China; 2 Yonsei University 

College of Medicine, Seoul, Korea (the Republic of); 3 Kaohsiung 

Medical University Hospital, Kaohsiung Medical University, 

Kaohsiung, Taiwan; 4 Istanbul University Cerrahpasa Faculty of 

Medicine, Istanbul, Turkey; 5 Liver Clinic, Surat, India; 6 University 

of Hamburg, Hamburg, Germany; 7 Gilead Sciences, Inc., Foster 

City, CA; 8 Yong Loo Lin School of Medicine, National University of 

Singapore, Singapore, Singapore; 9 University of Toronto, Toronto 

General Hospital, Toronto, ON, Canada; 10 Queen Marys University 

of London, London, United Kingdom; 11 Hospital Universitari 

Vall d’Hebron, Barcelona, Spain; 12 Second University of Naples, 

Naples, Italy; 13 The Chinese University of Hong Kong, Alice Ho 

Miu Ling Nethersole Hospital, Hong Kong, China; 14 Athens University 

Medical School, Athens, Greece; 15 Medical University of 

Bialystok, Bialystok, Poland; 16 Hôpital Beaujon, University of Paris, 

Paris, France 

Introduction: Combination treatment with pegylated interferon 

(PEG) and tenofovir disproxil fumarate (TDF) has increased 

rates of HBsAg loss as compared to either treatment alone. 

Here we evaluate the early HBsAg kinetics in patients treated 

with monotherapy (either PEG or TDF) or combination treatment 

of PEG+TDF. Methods: 740 CHB patients without advanced 

disease were randomized to receive (TDF+PEG) x48 weeks 

(arm A); (PEF+TDF) x16 weeks followed by TDF x32 weeks 

(arm B); TDF (arm C); PEG x48 weeks (arm D). Quantitative 

HBsAg was measured (Abbot Architect Assay) at baseline, 

Wks 4 and 12. Baseline predictors for HBsAg loss by Wk 72 

were evaluated using logistic regression analyses. Results: The 

proportion of patients with HBsAg declines of >0.5log 10 

, and 

>1log 10 

, at Wk 4 or 12 were higher among patients receiving 

PEG+TDF (arms A and B) as compared to monotherapy with 

PEG or TDF. By Wk 4, among patients receiving combination 

therapy (N=364), 5.5% had >1log 10 

decline and 11% 

had >0.5log 10 

decline, which increased by Wk 12 to 13.9% 

and 28.1% respectively (figure). Multivariate analyses of arms 

A+B showed that patients with >0.5log 10 

decline at Wk 4 or 

>1log 10 

decline at Wk 12 were more likely to be Genotype 

B and C with elevated ALT and HBV DNA. HBsAg declines in 

both monotherapy arms were also associated with similar baseline 

virologic characteristics. Within Arm A, sixteen patients 

experienced serum HBsAg loss by Wk 72. HBsAg declines 

of 1log 10 

at Wk 12 had a PPV of 0.31 and 0.43, 

respectively. Conclusions: Early kinetics of HBsAg with combination 

treatment of TDF and PEG as early as 4 weeks after 

treatment can predict eventual HBsAg loss with high NPV and 

modest PPV. The majority of patients treated, however, does 

Disclosures: 










George Wu - Employment: Gilead Bioscience 





















Janssen, BMS 








Robert Flisiak - Advisory Committees or Review Panels: Gilead, Bristol Myers 

Squibb, Janssen, Merck, Roche, Abbvie; Consulting: Janssen, Bristol Myers 

Squibb, Gilead, Merck, Abbvie; Grant/Research Support: Roche; Speaking and 

Teaching: Janssen, Roche, Bristol Myers Squibb, Gilead, Merck, Abbvie 





Abbvie 

The following authors have nothing to disclose: Sang Hoon Ahn, Fehmi Tabak, 

Rajiv Mehta, Aric Josun Hui


2015 

Safety and Efficacy of GS-4774 in Patients with Chronic 

Hepatitis B on Oral Antiviral Therapy 

Anna S. Lok 1 , Calvin Q. Pan 2 , Steven-Huy B. Han 3 , Huy N. 

Trinh 4 , Jeffrey Fessel 5 , Tim Rodell 6 , Benedetta Massetto 7 , Anh-Hoa 

Nguyen 7 , Anuj Gaggar 7 , Mani Subramanian 7 , John G. McHutchison 

7 , Carlo Ferrari 8 , Hannah Lee 9 , Stuart C. Gordon 10 , Edward 

J. Gane 11 ; 1 University of Michigan Health System, Ann Arbor, 

MI; 2 Medical Procare, PLLC, Flushing, NY; 3 Ronald Reagan UCLA 

Medical Center, Los Angeles, CA; 4 San Jose Gastroenterology, 

San Jose, CA; 5 Kaiser Permanente, San Francisco, CA; 6 Globeimmune, 

Louisville, CO; 7 Gilead Sciences, Inc., Foster City, CA; 8 University 

of Parma, Parma, Italy; 9 Tufts Medical Center, Boston, MA; 

10 Henry Ford Hospital, Detroit, MI; 11 Auckland General Hospital, 

Auckland, New Zealand 

Background: Chronic HBV infection is associated with a 

dysfunctional T-cell response. GS-4774 is a heat-inactivated 

yeast-based therapeutic T-cell vaccine expressing a recombinant 

protein containing HBV core, surface, and X proteins to 

elicit an HBV-specific T-cell response. This Phase 2 study evaluated 

GS-4774 in virally-suppressed chronic hepatitis B (CHB) 

patients. Methods: In this multicenter study, 178 non-cirrhotic 

patients with CHB were enrolled. All patients were virally suppressed 

on an oral antiviral (OAV) for >1 year. Patients were 

randomized to continue OAV alone or to continue OAV and 

receive GS-4774 every 4 weeks for 6 doses at either 2 yeast 

units (YU), 10 YU, or 40 YU [1 YU=10^7 yeast cells]. The 

primary endpoint was decline from baseline in quantitative 

serum HBsAg (Abbot Architect Assay) at Week 24. Safety 

assessments include monitoring of injection site reactions, 

hepatic flares, and virologic breakthrough. Results: Baseline 

demographics were similar across all groups. The mean age 

of patients was 45 to 50 years and 62-74% were men. The 

majority (68-80%) of patients was Asian, 24-26% were HBeAg 

positive, and the mean duration of prior OAV treatment was 

4.4-4.8 years. Grade 3 or 4 adverse events (AEs) occurred in 

30% 

reductions in HBsAg (observed variability in the OAV arm) 

at Week 48 (Panel B) at all timepoints. No patients achieved 

HBsAg loss. Five of 32 (16%) HBeAg positive patients treated 

with GS-4774 experienced HBeAg loss with 4 having HBeAg 

seroconversion. No virologic breakthrough was observed. Conclusions: 

GS-4774 was safe and well-tolerated in CHB patients 

with no patients achieving HBsAg loss during the study. Modest 

reductions in HBsAg were observed in some patients treated 

with GS-4774. 

Disclosures: 



Calvin Q. Pan - Advisory Committees or Review Panels: Gilead; Consulting: 

BMS, Gilead, Abbvie, Janssen ; Grant/Research Support: BMS, Gilead, Merck; 

Speaking and Teaching: BMS, Gilead, Abbvie 

Steven-Huy B. Han - Grant/Research Support: Bristol Myers Squibb, Gilead 




Jeffrey Fessel - Grant/Research Support: gilead, bms, abbvie, gsk, johnson & 

johnson 






Sciences 

Carlo Ferrari - Advisory Committees or Review Panels: Gilead, Roche, Abbvie, 

BMS, Merck, Arrowhead; Grant/Research Support: Gilead, Roche, Janssen 







The following authors have nothing to disclose: Tim Rodell, Anh-Hoa Nguyen, 

Hannah Lee 

2016 

Entecavir and Tenofovir More Effectively Reduce the 

Recurrence of Hepatitis B Virus-related Hepatocellular 

Carcinoma after Curative Treatment than Low-potent 

Antivirals 

Hongkeun Ahn 1 , Jeong-Hoon Lee 1 , YOUNG CHANG 1 , Joon Yeul 

Nam 1 , Hyeki Cho 1 , Young Youn Cho 1 , Minjong Lee 1 , Jeong-Ju 

Yoo 1 , Yuri Cho 1 , Donghyeon Lee 1 , Eun Ju Cho 1 , Su Jong Yu 1 , 

Yoon Jun Kim 1 , Jung-Hwan Yoon 1 , June Sung Lee 2 ; 1 Department of 

Internal Medicine and Liver Research Institute, Seoul National University 

Hospital, Seoul, Korea (the Republic of); 2 Internal Medicine, 

Inje University, Ilsan Paik Hospital, Goyang, Korea (the Republic 

of) 

Backgroud and Aim: The effect of antiviral potency of nucleos(t)ide 

analogues (NAs) on the recurrence of hepatocellular 

carcinoma (HCC) in chronic hepatitis B (CHB) patients has 

not been well investigated. This study aimed to evaluate the 

association of the potency of antivirals and the risk of hepatitis 

B virus (HBV)-related HCC recurrence after potentially curative 

treatment. Methods: We performed a retrospective analysis of 

data from 473 consecutive hepatitis B virus (HBV)-related HCC 

patients treated with radio-frequency ablation (RFA, n=261) 

or resection (n=212). According to the potency of antiviral 

treatments, total population was divided into three groups: 

group A (no antiviral treatment, n=224), group B (low-potent 

NA: telbivudine, adefovir, clevudine, and lamivudine; n=76), 

group C (high-potent NA: entecavir [ETV] and tenofovir disoproxil 

fumarate [TDF]; n=173). Recurrence-free survival (RFS) 

in three groups were analyzed by Kaplan-Meier curve analysis 

and Cox proportional hazards model. Results: The median 

follow-up duration was 56.9 months. The median RFS was 

38.3, 43.6, and 92.9 months in group A, group B, and group 

C, respectively (P=0.012 by log-rank test). In multivariate 

analysis, group C had significantly prolonged RFS (hazard 

ratio [HR]=0.55, 95% confidence interval [CI]=0.42–0.74; 

P


cant effect on RFS between groups B and C (HR=1.17, 95% 

CI=0.63–2.18; P=0.620). Conclusion: High-potent antiviral 

agents (ETV or TDF) reduce the risk of HCC recurrence compared 

to low-potent antivirals as well as no antivirals, especially 

in patients with high viral titer. 

Disclosures: 






The following authors have nothing to disclose: Hongkeun Ahn, Jeong-Hoon Lee, 

YOUNG CHANG, Joon Yeul Nam, Hyeki Cho, Young Youn Cho, Minjong Lee, 

Jeong-Ju Yoo, Yuri Cho, Donghyeon Lee, Eun Ju Cho, Su Jong Yu, Jung-Hwan 

Yoon, June Sung Lee 

2017 

Long term follow up of chronic hepatitis B patients after 

oral antiviral treatment discontinuation 

Seong Hee Kang, Jong Eun Yeon, Young-Sun Lee, Tae Suk Kim, 

Yang Jae Yoo, Ji Hoon Kim, Kwan Soo Byun; Gastroenterology & 

Hepatology, Korea University Medical Center, Seoul, Korea (the 

Republic of) 

Introduction: In the current guidelines, 6-12 months of consolidation 

after the HBeAg seroconversion is recommended for 

HBeAg-positive patients. And the ideal treatment duration of 

HBeAg negative chronic hepatitis B (CHB) is not well known. 

We evaluated long term clinical courses in patient after antiviral 

treatment cessation. Methods: A total of 118 HBeAg-positive 

and 60 HBeAg-negative CHB patients who discontinued 

lamivudine between 1997 and 2014 were retrospectively analyzed. 

Results: In HBeAg-positive patients, the mean age of the 

patients at treatment initiation was 32.5 years. The mean duration 

of lamivudine treatment in was 32.3 months and the mean 

follow-up period after discontinuation was 72.4 months. The 

cumulative probability of a composite relapse (≥ 10 4 copies/ 

mL) at 1, 6, 12, 24, 48, 96, 180 months were 11.9%, 28.8%, 

38.1%, 51.7%, 59.3%, 63.6%, 64.4% respectively. The mean 

duration of lamivudine treatment were 31.9 months and 32.9 

months in the relapse and non-relapse group (p=0.79). The 

mean duration of consolidation period after HBeAg seroconversion 

were 10.7 months and 11.4 months were not different 

(p=0.792). Multivariate analysis revealed pretreatment age ≤ 

34 years [HR 0.324, 95% CI: 0.148-0.710 p=0.005] and 

undetectable HBV DNA [HR 0.261, 95% CI: 0.107-0.638 

p=0.003] within the three month after treatment discontinuation 

were the predictive factors of non-relapse. In HBeAg-negative 

patients, the mean age of the patients was 39.2 years. 

The cumulative probability of a composite relapse at 1, 6, 12, 

24, 48, 96, 180 months were 25.0%, 33.3%, 35.0%, 41.7%, 

43.3%, 46.7%, 48.3% respectively. Mean time to relapse after 

off-treatment was 13.6 months. The mean duration of lamivudine 

treatment were 29.8 months and 33.5 months in the 

relapse and non-relapse group (p=0.447). Conclusions: Even 

with the long term consolidation after HBeAg seroconversion, 

most HBeAg positive CHB patients showed relapse. And half 

of patients with HBeAg-negative CHB had relapsed after treatment 

cessation. Antiviral therapy should be maintained for long 

term period or until HBsAg was lose. 

Disclosures: 


The following authors have nothing to disclose: Seong Hee Kang, Jong Eun Yeon, 

Young-Sun Lee, Tae Suk Kim, Yang Jae Yoo, Ji Hoon Kim 

2018 

The impact of timely initial injection of two different 

doses of hepatitis B virus (HBV) vaccine plus hepatitis B 

immunoglobulin (HBIG) on preventing perinatal transmission 

of HBV infection 

chong wang 1 , Chuan Wang 2 , Jie Li 3 , xing Wu 2 , Fei Kong 1 , Simin 

Wen 2 , Jing Jiang 2 , Junqi Niu 1 ; 1 hepatology, The First Hospital of 

Jilin University, Changchun, China; 2 The First Hospital of Jilin University, 

Changchun, China; 3 Department of microbiology, Beijing 

university, Beijing, China 

Background & aims: Routine immunoprophylaxis reduces HBV 

transmission. However, it does not have individual vaccine 

dose to babies whose mothers with different hepatitis B virus e 

antigen (HBeAg) status. this study was to investigate the effectiveness 

of an immunoprophylaxis protocol with two different 

vaccine doses according to mothers’HBeAg(+/-) status and 

timely initial injection on blocking mother-to-infant transmission 

of HBV, and to evaluate potential risk factors associated with 

immunoprophylaxis failure and protective antibody titers. Methods: 

A population-based prospective study was conducted from 

July 2012 to April 2015. A total of 863 hepatitis B virus surface 

antigen(HBsAg)-positive mothers and their 871 infants (8 

pairs of twins) were included in the study. The initial injection 

of HBV vaccine and HBIG(100IU) were carried out within 2 

hours of birth. 20mg and 10mg HBV vaccine were given to 

babies born to HBeAg+ and HBeAg- mothers respectively. We 

analyze the association of multiple risk factors with immunoprophylaxis 

failure and vaccine response. Results: At 7 months, 

no immunoprophylaxis failure was observed in 565 babies 

born to HBeAg- mothers.Among 306 infants born to HBeAg+ 

mothers, 16 (5.2%) babies were immunoprophylaxis failure 

(HBsAg-positive) at 7 months, with high HBV DNA load ≥8 

log10 IU/ml (Crude OR:3.69, 95%CI:1.03-13.24; Adjusted 

OR:4.53, 95%CI a :1.19-17.34; P a =0.027) and Non timely 

or inadequate initial injection (Crude OR:3.42, 95%CI:1.12- 

10.49; Adjusted OR:5.23, 95%CI a :1.54-17.82; P a =0.008) 

were strongly associated with immunoprophylaxis failure, 

while other factors including delivery mode, feeding pattern, 

infant gender and birth weight showed no significant association. 

Of 855 babies for whom blocking was successful, 

825(96.5%) had produced adequate titers of antibody to 

HBsAg (anti-HBs) (≥ 100 mIU/mL). For full-term babies, birth 

weight< 3000 g was correlated with low immune response 

to vaccination (Crude OR:0.37, 95%CI:0.17-0.84; Adjusted 

OR:0.37, 95%CI a :0.16-0.84; P a =0.018). Conclusions: Application 

of timely initial injection of two different doses of HBV 

vaccine plus HBIG to infants born to mothers with different 

HBeAg status is effective for preventing perinatal transmission 

of HBV, particularly for those whose mothers are HBeAg-negative. 

The overwhelming majority of babies produced adequate 

levels of protective anti-HBs titers (>100 mIU/mL) after immunoprophylaxis. 

high HBV DNA load, and non-timely or inadequate 

initial injection were associated with immunoprophylaxis 

failure. Relative low birth weight of full-term babies might be 

correlated with weak immune response to vaccination. 

Disclosures: 

The following authors have nothing to disclose: chong wang, Chuan Wang, Jie 

Li, xing Wu, Fei Kong, Simin Wen, Jing Jiang, Junqi Niu


2019 

Discontinuation of Effective Long-Term Nucleos(t)ide 

Analogue(s) [NA(s)] Treatment in HBeAg-Negative 

Chronic Hepatitis B (CHBe-) Patients: A Multicenter 

Ongoing Prospective Study 

Spilios Manolakopoulos 1 , Hariklia Kranidioti 1 , Anastasia 

Kourikou 1 , Emilia Hadziyannis 1 , Georgios Kontos 1 , Melanie 

Deutsch 1 , Alexandra Alexopoulou 1 , Emanuel K. Manesis 1 , George 

V. Papatheodoridis 2,1 ; 1 2nd Department of Internal Medicine, University 

of Athens, Hippokration General Hospital, Athens, Greece; 

2 Department of Gastroenterology, University of Athens, Laiko General 

Hospital, Athens, Greece 

Background/Aim The durability of response to NA(s) after 

their cessation, has not been adequately investigated in 

CHBe- patients. This prospective study aims to investigate the 

outcome of CHBe- patients who discontinue NA(s) after longterm 

virological suppression and to explore factors that may 

predict relapse. Methods Non-cirrhotic CHBe- patients with 

virological remission (undetectable HBV-DNA) under NA(s) 

for ≥4 years who could remain under close follow-up (every 

month for the first 3 months and every 3 months thereafter) 

were invited to consent to treatment discontinuation. Τhe criteria 

for NA(s) retreatment were: ALT>2xULN & Bilirubin>2mg/ 

dl, ALT>10xULN, ALT>5xULN in 2 monthly determinations, 

ALT>3xULN and HBV-DNA>100,000IU/mL, ALT>ULN & HBV- 

DNA>2,000IU/mL for ≥6 months. Results Until May 2015, 

43 patients [males: 74%, median age: 60 (22-86) ys, duration 

of NA(s): 7.9 (4-14) ys] had >6 months follow-up [median: 27 

(6-45) months] and were included in the analysis. Off-treatment 

virological relapse (HBV-DNA >2000IU/mL) was developed 

in 40 (93%) and biochemical relapse (ALT >ULN) in 29 (67%) 

patients. Ten patients (23%) experienced hepatitis flare (ALT 

>10 ULN) at a median of 3 (1-9) months after NA(s) discontinuation, 

without evidence of liver decompensation. Retreatment 

was required in 18 (42%) patients at a median of 6.5 (1-33) 

months. Of the 25 patients who remained untreated, 14 (56%) 

had ALT persistently


The following authors have nothing to disclose: Tetsuya Hosaka, Masahiro 

Kobayashi, Shunichiro Fujiyama, Hideo Kunimoto, Yushi Sorin, Yusuke 

Kawamura, Hitomi Sezaki, Satoshi Saitoh, Yasuji Arase, Mariko Kobayashi 

2021 

Antiviral Activity of Tenofovir Alafenamide (TAF) Against 

Drug Resistant HBV Isolates In Vitro 

Yang Liu, Ben C. Mitchell, Phillip Dinh, Michael D. Miller, Kathryn 

M. Kitrinos; Gilead Sciences, Foster City, CA 

Background: Tenofovir alafenamide (TAF) is a next generation 

prodrug of tenofovir that is being evaluated for the treatment 

of hepatitis B virus (HBV) and human immunodeficiency virus 

(HIV) infections. The aim of this study was to evaluate the in 

vitro antiviral activity of TAF against adefovir-resistant (ADV- 

R), lamivudine-resistant (LAM-R), and entecavir-resistant (ETV- 

R) isolates. Methods: A full length genotype D HBV genome 

was amplified from a treatment-naïve subject and cloned 

into an expression vector under the direction of a CMV promoter. 

Eleven site-directed mutants were created: 5 ADV-R 

(rtA181T/sW172*, rtA181T/sW172L, rtA181V, rtN236T, 

rtA181V+N236T), 3 LAM-R (rtM204I, rtL180M+rtM204V, 

rtV173L+rtL180M+rtM204V), and 3 ETV-R (rtL180M+rtM- 

204V+rtT184G, rtL180M+rtM204V+rtS202G, rtL180M+rt- 

M204V+rtM250V) mutants. All constructs were transiently 

transfected into HepG2 cells and treated with TAF and tenofovir 

(TFV). Drug susceptibility was assessed using quantitative 

PCR of intracellular HBV DNA. Fold change (FC) values were 

generated by comparing the average EC 50 

values obtained 

for the mutant viruses to the wild-type virus. Results: Transient 

transfection of all constructs resulted in efficient HBV DNA replication 

in HepG2 cells. The TAF susceptibility for all the drug 

resistant isolates demonstrated mean EC 50 

values ranging from 

85.4 to 364.7 nM, while the wild-type TAF EC 50 

was 99.1 

nM. The mean FC values for the drug resistant isolates ranged 

from 0.9-3.7, with all but one isolate remaining sensitive to TAF 

(FC < 2). The ADV-R isolate rtA181V+rtN236T demonstrated 

low-level TAF reduced susceptibility (FC = 3.7). The TFV susceptibility 

for all the drug resistant isolates demonstrated mean 

EC 50 

values ranging from 7.8 to 40.5 mM, the mean FC values 

for the drug resistant isolates ranged from 0.6 to 2.8, with all 

but one isolate remaining sensitive to TFV (FC < 2). The ADV-R 

isolate rtA181V+rtN236T demonstrated low-level reduced susceptibility 

to TFV (FC = 2.8). Despite the difference in EC 50 

values, 

the FC values obtained for each isolate with TAF and TFV 

were consistent. Conclusions: All the ADV-R, LAM-R and ETV-R 

mutant isolates were sensitive to TAF with the exception of the 

ADV-R double mutant rtA181V+rtN236T, which exhibited low 

level reduced susceptibility to TAF. The FC values observed 

with TAF are similar to what has been previously observed with 

TFV. These data supports the on-going TAF phase 3 clinical 

trial program in treatment naive and treatment experienced 

HBV patients. 

Disclosures: 

Yang Liu - Employment: Gilead Sciences 





Sciences 

The following authors have nothing to disclose: Ben C. Mitchell 

2022 

Monthly dosing of ARC-520 in chronically hepatitis B 

virus infected chimpanzees produces rapid, deep and 

durable reductions in circulating viral antigens 

Christine I. Wooddell 1 , Deborah Chavez 3 , Jason E. Goetzmann 2 , 

Lena M. Notvall-Elkey 3 , Helen Lee 3 , Bernadette Guerra 3 , Ryan M. 

Peterson 1 , Courtney N. Johnson 3 , Julia O. Hegge 1 , Robert Gish 4 , 

Stephen Locarnini 5 , Christopher R. Anzalone 1 , Robert E. Lanford 3 , 

David L. Lewis 1 ; 1 Arrowhead Madison, Arrowhead Research Corporation, 

Madison, WI; 2 New Iberia Research Center, University 

of Louisiana at Lafayette, New Iberia, LA; 3 Texas Biomedical 

Research Institute, San Antonio, TX; 4 Department of Medicine, 

Stanford University Medical Center, San Diego, CA; 5 Victorian 

Infectious Diseases Reference Laboratory, Melbourne, VIC, Australia 

Background: New therapeutic strategies are needed for 

chronic HBV infection. RNAi-based therapeutic ARC-520 that 

was designed to degrade viral transcripts and reduce HBV 

antigens and pgRNA is in clinical development. This study 

assessed multiple doses of ARC-520 in HBV infected chimps. 

Methods: 9 chimps (5 M, 4 F; 9-37 yrs) with chronic HBV were 

treated with NUCs for 8-24 weeks prior to receiving ARC- 

520. Following the NUC lead-in, ARC-520 was added and 

dosed once every 4 weeks (Q4W) for 6-11 doses. 5 chimps 

were HBeAg-positive (HBeAg+) and 4 were HBeAg-negative 

(HBeAg-) at start of study. Chimps received humane care as 

outlined in the Guide for the Care and Use of Laboratory Animals. 

Results: Prior to treatment, HBV serum DNA levels ranged 

from 1.1E+7 - 7.1E+8 IU/mL in HBeAg+ chimps and 20-660 

IU/mL in HBeAg- chimps. HBsAg levels ranged from 250-3190 

μg/mL in HBeAg+ chimps and from 1.2–200 μg/mL in HBeAgchimps. 

After the NUC lead-in, DNA levels were reduced to 

≤10 3 IU/mL in HBeAg+ chimps and to undetectable levels in 

HBeAg- chimps. HBsAg levels were only marginally decreased. 

All 9 chimps were then treated with 2, 3 or 4 mg/kg ARC-520. 

All procedures and treatments were well tolerated. Treatment 

initiation with ARC-520 resulted in strongly reduced viral antigens 

with the magnitude of the response in HBeAg+ chimps 

being greater than in HBeAg- chimps. Following the first injection 

of ARC-520 in HBeAg+ chimps, HBsAg was reduced at 

nadir by 0.5-1.4 log 10 

, while in HBeAg- chimps HBsAg was 

reduced by 0.4–0.7 log 10 

. Doses of 2, 3 and 4 mg/kg had 

similar efficacy. Levels of HBsAg tended to decrease upon further 

dosing. Following the final dose, HBsAg showed mean 

reduction at nadir of 1.7 ± 0.5 log 10 

in HBeAg+ chimps and 

0.7 ± 0.1 log 10 

in HBeAg- chimps. HBeAg in HBeAg+ chimps 

was reduced by 1.5– 2.0 log 10 

. Two HBeAg+ chimps seroconverted 

to HBeAg-, anti-HBe positive. Four months after all therapies 

ended, one of these has maintained 0.3 log 10 

reduction in 

HBsAg and greater than 2.0 log 10 

reduced serum DNA relative 

to baseline; the other has maintained 1.4 log 10 

reduced HBsAg 

and greater than 4.0 log 10 

reduced serum DNA. The latter had 

an on-treatment ALT flare that resolved. Conclusions: ARC-520 

administered Q4W on a background of daily NUCs was well 

tolerated and produced large reductions in circulating HBV 

antigens in chimpanzees. Responses were greater in HBeAg+ 

chimps compared to HBeAg-; 2/5 HBeAg+ chimps serocleared 

HBeAg. ARC-520 administered to HBV infected chimpanzees 

produces rapid, deep and durable reductions in circulating 

HBsAg and HBeAg when dosed over a short defined period. 

Disclosures: 

Christine I. Wooddell - Employment: Arrowhead Research Corporation 

Ryan M. Peterson - Employment: Arrowhead Research Corporation 

Julia O. Hegge - Employment: Arrowhead Research Corp






Health 

Robert E. Lanford - Grant/Research Support: Arrowhead Research 

David L. Lewis - Employment: Arrowhead Research Corporation 

The following authors have nothing to disclose: Deborah Chavez, Jason E. Goetzmann, 

Lena M. Notvall-Elkey, Helen Lee, Bernadette Guerra, Courtney N. Johnson, 

Christopher R. Anzalone 

2023 

Efficacy and Safety of Peginterferon alfa-2b (40kD, 

Y Shape) (Pegberon) in HBeAg- positive CHB Patients 

—- A Phase III, Randomized, Multi-center, Positive-Controlled, 

Open-label Clinical Study 

Gui-Qiang Wang 1 , Feng-Qin Hou 1 , Wei Lu 2 , Jia Shang 3 , Guo- 

Zhong Gong 4 , Chen Pan 5 , Ming-Xiang Zhang 6 , Chi-Biao Yin 7 , 

Qing Xie 8 , Yan-Zhong Peng 9 , Shi-Jun Chen 10 ; 1 Peking University 

First Hospital, Beijing, China; 2 Xiamen Amoytop Biotech Co.,Ltd, 

Xiamen, China; 3 Henan Provincial People’s Hospital, Zhengzhou, 

China; 4 The Second Xiangya Hospital of Central South University, 

Changsha, China; 5 Mengchao Hepatobiliary Hospital of Fujian 

Medical University, Fuzhou, China; 6 The Sixth People’s Hospital 

of Shenyang, Shenyang, China; 7 Guangzhou Eighth People’s 

Hospital, Guangzhou, China; 8 Rui Jin Hospital, Shanghai, China; 

9 Peking University Shenzhen Hospital, Shenzhen, China; 10 Jinan 

Infectious Diseases Hospital, Jinan, China 

Background: Chronic hepatitis B (CHB) is a life-threatening 

liver disease caused by the hepatitis B virus in the world including 

China. Peginterferon is one of the first-line drugs recommended 

for CHB in guidelines. Thusit is necessary to develop 

new peginterferon in China and construct higher-level basis of 

evidence-based medicine in Chinese CHB patients. Objective: 

Evaluate efficacy and safety of Pegberon (180μg), that developed 

in Chinese domestic company with gobal patent 40kD 

Y-shape peglated interferon α-2b, by the head-to-head, randomized, 

multi-center, positive controlled and open label clinical 

study with PEG IFNα-2a (180μg). Provide a higher level 

basis of evidence-based medicine for peginterferon treatment. 

Methods: Open-label Phase III study enrolled 538 patients for 

Pegberon group and 282 patients for PEG IFNα-2a group 

with 48 weeks treatment and 24 weeks follow up. The proportion 

of patients with HBeAg seroconversion and the safety 

were evaluated. HBV DNA analyzed by COBAS ® , Version 

2.0. Serum markers including quantitative HBsAg and HBsAb 

were analyzed by cobas e 411 analyzer, and HBeAg was 

analyzed by Elecsys HBeAg, HBeAb was analyzed by Elecsys 

anti-HBe. Results: Pegberon group and PEG IFNα-2a group 

had similar baseline virological and serological characteristics. 

30.82%(102/331) patients in Pegberon group and 

27.27%(48/176) patients in PEG IFNα-2a group achieved 

HBeAg seroconversion respectively in the 507 patients that 

completed the 72 weeks clinical trial at present. In intent-to 

treat populationsboth groups had similar trends of HBV DNA, 

HBeAg and HBsAg during 48-week treatment. At 48w, HBV 

DNA negative rate in Pegberon group was 11.90%(64/538) 

while 9.93%(28/282) in PEG IFNα-2a group. In addition, 

rate of HBV DNA10 6 IU/mL enrolled were 

treated with antiviral therapy from 24-32 weeks of gestation 

using NUCs,except for LDT+TDF at the first trimester, including 

330 in LdT group, 25 in TDF group, 27 in TDF+LdT group due 

to LdT resistance, respectively. 171 cases who were unwilling 

to take antiviral drugs served as controls. All infants were 

vaccinated with recombinant HBV vaccine and hepatitis B 

immune globulin (HBIG) according to standard immunoprophylaxis 

procedure. MTCT rate was determined by HBV markers 

including HBsAg and HBV DNA detection in 6 months after 

birth. Results: Significantly lower HBV DNA levels were noted 

in all the enrolled mothers when delivery. The rate of HBsAg 

positive and HBV DNA detectable in infants at 6 months was 

0% in each group. The incidence of undetectable cord blood 

HBV DNA levels has no significant difference among the three 

groups (LdT group, 99.2%; TDF group, 100%, LdT+TDF group, 

100%, P>0.05),which were significantly higher than the controls(61.5%),No 

severe adverse events or complications were 

observed in all the mothers and infants. Conclusions: LdT and 

TDF monotherapy or combotherapy were all effective and 

well-tolerated in HBeAg positive high viral load mothers and 

their infants on short term follow up, and these were associated 

with significant reduction of MTCT. 

Disclosures: 

The following authors have nothing to disclose: yanqiong zhang, Hongfei Huang, 

Quanxin Wu, Yuming Wang


2025 

Descriptive Comparison of Efficacy and Safety of Tenofovir 

Disoproxil Fumarate (TDF) plus Pegylated-Interferon 

α-2a (PEG) Combination in Asians and non-Asians 

with Chronic Hepatitis B (CHB) 

Patrick Marcellin 2 , Sang Hoon Ahn 3 , Won Young Tak 4 , Owen 

Tak Yin Tsang 8 , Aric Josun Hui 5 , Xiaoli Ma 6 , Wan-Long Chuang 7 , 

Seng Gee Lim 9 , Rajiv Mehta 10 , Eduardo B. Martins 1 , Leland J. 

Yee 1 , Phillip Dinh 1 , Lanjia Lin 1 , Prista Charuworn 1 , Mani Subramanian 

1 , Scott Fung 11 , Magdy Elkhashab 12 , Huy N. Trinh 13 , Henry 

Lik-Yuen Chan 14 ; 1 Gilead Sciences, Foster City, CA; 2 Hôpital 

Beaujon, Université Paris-Diderot, Clichy, France; 3 Yonsei University 

College of Medicine, Seoul, Korea (the Republic of); 4 Kyungpook 

National University Hospital, Daegu, Korea (the Republic 

of); 5 Alice Ho Miu Ling Nethersole Hospital, Hong Kong, China; 

6 Drexel University College of Medicine, Philadelphia, PA; 7 Kaohsiung 

Medical University Chung-Ho Memorial Hospital, Kaohsiung, 

Taiwan; 8 Princess Margaret Hospital, Hong Kong, China; 9 Yong 

Loo Lin School of Medicine, National University of Singapore, Singapore, 

Singapore; 10 Liver Clinic, Surat, India; 11 Toronto General 

Hospital, Toronto, ON, Canada; 12 Toronto Liver Centre, Toronto, 

ON, Canada; 13 San Jose Gastroenterology, San Jose, CA; 14 The 

Chinese University of Hong Kong, Hong Kong, China 

Background: Differences in viral and patient characteristics 

between Asians and non-Asians may impact response to HBV 

therapy. We examined differences in between Asian and 

non-Asian patients enrolled in the TDF plus PEG combination 

study GS-US-174-0149. Methods: 740 CHB patients without 

advanced liver disease were randomized 1:1:1:1 to receive 

TDF+PEG x48 weeks (Arm A); TDF+PEG x16 weeks followed 

by TDF x32 weeks (Arm B); continuous TDF (Arm C); PEG x48 

weeks (Arm D). Baseline characteristics, Week 72 efficacy and 

overall safety were compared between Asians and non-Asians. 

Primary efficacy was HBsAg loss at Week 72. Results: Of 554 

Asian patients randomized, most were male and HBeAg-positive, 

and predominantly genotype (Gt)-B/C; while among 

186 non-Asian patients, most were male, HBeAg-negative and 

predominantly Gt- A/ D (Table 1). Overall, rates of HBsAg 

decline ≥ 1 log 10 

IU/mL from baseline to Week 48 were lower 

among non-Asians (11%) vs. Asians (26%) (p0.05 for 

Asian vs non-Asian comparison within each treatment group). 

Within each treatment arm, no significant differences were 

observed for rates of normal ALT, HBeAg loss/seroconversion 

between Asian and non-Asian patients at Week 48. Rates of 

treatment-emergent AEs, discontinuations and SAEs were similar 

between Asians and non-Asians. Conclusion: Although more 

Asians had >1log 10 

HBsAg decline from baseline, no statistically 

significant differences in efficacy were observed between 

Asians and non-Asians. 

Table 1 

*Among 497 Asian and 162 Non-Asians who reached Week 

48. 

N.S.=Not Significant 

Disclosures: 





Abbvie 



Xiaoli Ma - Consulting: Gilead Sciemces, Inc 












Lanjia Lin - Employment: Gilead; Stock Shareholder: Gilead 

Prista Charuworn - Stock Shareholder: Gilead Sciences 




Magdy Elkhashab - Advisory Committees or Review Panels: GSK, Gilead, Merk, 

BMS, ABBVIE; Grant/Research Support: EISAI, GENENTECH, GSK, GILEAD, 

ABBVIE, Merk, BMS 







The following authors have nothing to disclose: Sang Hoon Ahn, Owen Tak Yin 

Tsang, Aric Josun Hui, Rajiv Mehta 

2026 

Characterization of Hepatitis B Surface Proteins in 

HBeAg-positive Patients with Chronic Hepatitis B (CHB) 

Treated With PegInterferon Alfa-2a (40KD) 

Franziska Rinker 1,2 , Corinna M. Bremer 3,4 , Steffen B. Wiegand 1 , 

Birgit Bremer 1 , Michael P. Manns 1,2 , Heiner Wedemeyer 1,2 , Lei 

Yang 5 , Vedran Pavlovic 6 , Cynthia Wat 6 , Dieter Glebe 3,4 , Anke R. 

Kraft 1 , Markus Cornberg 1,2 ; 1 Gastroenterology, Hepatology and 

Endocrinology, Medical School Hannover, Hannover, Germany; 

2 German Center for Infection Research (DZIF), Hannover, Germany; 

3 Institute of Medical Virology, National Reference Center 

for Hepatitis B and D Viruses, Justus Liebig University Giessen, 

Giessen, Germany; 4 German Center for Infection Research (DZIF), 

Giessen, Germany; 5 Roche Products Ltd, Shanghai, China; 6 Roche 

Products Ltd, Welwyn Garden City, United Kingdom 

Background: HBsAg consists of three proteins called large 

(L-), middle (M-) and small (S-)HBs that differ in amino-terminal 

sequences and glycosylation status. Commercial HBsAg assays 

only quantify the total amount of HBsAg. We have validated 

an ELISA method for quantification of HBs proteins and demonstrated 

that SHBs represents the commonest subtype (accounting 

for 88% of total HBsAg, L- and M- HBs account for 8% 

and 3%) in untreated HBeAg-positive patients. The aim of current 

study was to explore HBs protein fractions kinetics during 

pegylated interferon alfa-2a (PegIFNα-2a, Pegasys) therapy 

and examine whether HBs fractions are more predictive of 

treatment response than quantitative HBsAg (qHBsAg) levels. 

Methods: Serum samples of 128 HBeAg-positive patients, from 

two phase III/IV PegIFNα-2a trials were retrospectively analysed. 

Patients received PegIFNα-2a ± lamivudine therapy for 

48 weeks. Clinical and laboratory patient data from completed 

trials was provided by Roche. HBs fractions were quantified


using a quantitative ELISA and well-defined monoclonal antibodies 

against the S-domain (Total_HBs), the preS1-domain 

(LHBs) and the N-glycosylated preS2-domain (MHBs) of the HBs 

protein. SHBs levels were derived by subtracting L- and M- HBs 

from Total_HBs. Data was log-transformed (log 10 

x+1) prior to 

analysis. Response was defined as HBeAg seroconversion 24 

weeks post-treatment. Results: During treatment, mean HBs fractions 

and qHBsAg levels were consistently lower in responders 

than non-responders. HBs fractions declined by approximately 

1 log by Week 24 (range 0.9-1.2), which was comparable 

to the 0.9 log decline observed in qHBsAg levels. Receiver 

operating characteristics (ROC) analysis demonstrated that 

HBV-DNA, qHBsAg and qHBeAg (but not ALT) levels can predict 

treatment response moderately well with area under the 

ROC curve (AUROC) scores of 0.70-0.86 (see Table). AUROC 

scores for HBs fractions (range 0.70-0.75) were comparable 

to qHBsAg scores (0.70-0.74) at each time-point. Conclusions: 

The results demonstrate that during PegIFNα-2a therapy, HBs 

fractions are lower in responders than non-responders, which 

is consistent with the pattern observed with qHBsAg levels. The 

ROC analysis demonstrates that individual HBs fraction levels 

are equivalent and not superior to qHBsAg levels, for predicting 

treatment response in HBeAg-positive patients. 

AUROC Scores at Baseline, Week 12 and 24 

2027 

Liver Gene Expression Profiles Correlate with Virus 

Infection and Response to Interferon Therapy in Chronic 

Hepatitis B Patients 

Chun-Jen Liu, Pei-Jer Chen, Ding-Shinn Chen, Jia-Horng Kao, Hui- 

Lin Wu; Graduate Institute of Clinical Medicine, National Taiwan 

University College of Medicine, Taipei City, Taiwan 

Background: The natural course of chronic hepatitis B (CHB) 

infection and treatment response are determined mainly by the 

genomic characteristics of the individual. We investigated liver 

gene expression profiles to reveal the molecular basis associated 

with viral infection and IFN-alpha treatment response 

in CHB patients. Methods: Nineteen IFN-alpha responders 

and 19 non-responders with CHB were included in this study. 

Expression profiles were compared between paired liver 

biopsy samples taken before and 6 months after successful 

IFN-alpha treatment or between pretreatment biopsy samples 

of IFN-alpha responders and non-responders. Results: A 

total of 132 differentially up-regulated and 39 differentially 

down-regulated genes were identified in the pretreatment livers 

of CHB patients. The up-regulated genes were mainly related 

to immune response and cell proliferation. IFN-alpha and B cell 

signatures were also enriched. Lower intrahepatic HBV pregenomic 

RNA levels and a total of 118 differentially up-regulated 

and 33 differentially down-regulated genes were identified in 

IFN-alpha responders. The up-regulated gene set in responders 

significantly overlapped with the up-regulated genes associated 

with the pretreatment livers of CHB patients. In contrast, 

no clear pattern was found in the down-regulated genes of 

both analyses. Results of the bioinformatic analyses were validated 

in independent cohorts and relevant animal models. 

Conclusions: CHB infection evokes significant gene expression 

associated with immune responses. The up-regulated genes are 

predictive of responsiveness to IFN-alpha therapy, as are lower 

intrahepatic levels of HBV pregenomic RNA and pre-activated 

host immune responses. Our findings provide novel insights 

into the immune mechanism and manipulation of CHB. 

Disclosures: 

Pei-Jer Chen - Advisory Committees or Review Panels: BMS, GSK, BMS, GSK, 

Medigene; Independent Contractor: J & J; Speaking and Teaching: Roche, Roche 

The following authors have nothing to disclose: Chun-Jen Liu, Ding-Shinn Chen, 

Jia-Horng Kao, Hui-Lin Wu 

*HBV-DNA analyses performed on PEgIFN monotherapy cohort 

Disclosures: 





Novartis 





Lei Yang - Employment: Roche (China) Holding Ltd. 

Vedran Pavlovic - Employment: Roche Products Ltd; Stock Shareholder: Roche 

Products Ltd 

Cynthia Wat - Employment: Roche Products Ltd 





The following authors have nothing to disclose: Franziska Rinker, Corinna M. 

Bremer, Steffen B. Wiegand, Birgit Bremer, Dieter Glebe, Anke R. Kraft 

2028 

A study to evaluate the dynamics changes of HBsAg 

quantity and its relation with HBeAg seroconversion following 

48 weeks pegylated-interferon-alpha treatment 

in patients with HBeAg positive chronic hepatitis B after 

long term nucleos(t)ide analogue maintenance therapy 

(Roll Over trial) : Interim analysis at 24 weeks 

Jeong Heo 1 , Hyun Young Woo 1 , Won Young Tak 2 , Soo Young 

Park 2 , Won Lim 1 , Youngmi Hong 1 , Young Oh Kweon 2 , Ki Tae 

Yoon 1 , Mong Cho 1 ; 1 Pusan National University Hospital, Pusan, 

Korea (the Republic of); 2 Kyungpook National University School of 

Medicine, Daegu, Korea (the Republic of) 

Background & Aims: Durable post-treatment response is uncommon 

in patients with chronic hepatitis B (CHB) on nucleos(t) 

ide analogue (NA) therapy. The aim of this study is to investigate 

pegylated interferon (PI) after long term NA therapy might 

potentiate the antiviral efficacy directly via its effect on broad 

antiviral activities and indirectly via activation of innate and 

adaptive immune responses leading to HBeAg seroconversion 

and eventually HBsAg loss and/or seroconversion. Methods: 

The patient with HBeAg-positive CHB who had been treated 

with any NA except telbivudine, who have an undetectable 

HBV DNA (


However, none of these on treatment HBV DNA elevation was 

accompanied by elevated level of alanine aminotransferase 

(ALT). The level of baseline ALT showed association with HBeAg 

seroconversion (p=0.075). PI alfa-2a was well-tolerated. Conclusions: 

This interim analysis showed that, for patients who 

achieve virological suppression with oral NA, switching to a 

24 weeks PI alfa-2a treatment significantly decrease HBsAg 

titer and increases rates of HBeAg seroconversion. 

Disclosures: 

Jeong Heo - Advisory Committees or Review Panels: Abbvie; Grant/Research 




Ki Tae Yoon - Grant/Research Support: Handok; Speaking and Teaching: Gilead, 

BMS, MSD, Roche, GSK 

The following authors have nothing to disclose: Hyun Young Woo, Soo Young 

Park, Won Lim, Youngmi Hong, Young Oh Kweon, Mong Cho 

2029 

Reduction of HBsAg by peg-interferon given sequentially 

after long term nucleot(s)ide analogue therapy: 

nationwide multicenter study by Japanese Red Cross 

Liver Study Group. 

Masayuki Kurosaki, Namiki Izumi; Department of Gastroenterology 

and Hepatology, Musashino Red Cross Hospital, Tokyo, Japan 

Background and Aims: The ideal endpoint of antiviral therapy 

for chronic hepatitis B is loss of HBs antigen (HBsAg). However 

HBsAg loss is rarely achieved by nucleot(s)ide analogues 

(NUC). The aim of the present study is to elucidate the degree 

of HBsAg reduction and incidence of HBsAg loss by peg-interferon 

when given sequentially after long term NUC therapy. 

Methods: A total of 54 patients were sequentially treated by 48 

weeks of peg-interferon alpha-2a (180 microgram weekly) after 

long-term NUC therapy with an overlap of 4 weeks (sequential 

therapy group). Preceding NUC therapy was entecavir in 85% 

and lamivudine plus adefovir in 15% of patients. The average 

duration of preceding NUC therapy was 4.1 years. At the 

start of peg-interferon, HBV DNA was undetectable in 93%, 

and HBeAg was negative in 69% of patients. For comparison, 

68 patients treated by 48 weeks of peg-interferon alpha-2a 

(180 microgram weekly) without preceding NUC therapy 

was studied (monotherapy group). Serial changes in the titer 

of HBsAg were measured by quantitative assay. Results: In 

sequential therapy group, HBV DNA remained undetected in 

80% of patients after start of peg-interferon and withdrawal of 

NUC. In HBeAg positive patients, HBeAg became negative at 

the end of therapy in 50% in sequential therapy and 14% in 

monotherapy (p=0.04). At the start of peg-interferon, titer of 

HBsAg was 850 (1.9-36500) IU/mL in sequential therapy and 

1990 (6.0-80360) IU/mL in monotherapy which declined to 

166 (1.0 Log IU/mL was 34% vs. 19%, HBsAg reduction 

>2.0 Log IU/mL was 16% vs. 7%, and HBsAg loss was 6% 

vs. 0% for sequential and monotherapy, respectively. Within 

the same individual, average HBsAg reduction during sequential 

peg-interferon was significantly higher compared to those 

during preceding NUC therapy (0.64 vs. 0.16 Log/year, 

p=0.015). Among patients with HBsAg 60 IU/L and decrease in HBsAg level 

1.0 Log IU/mL during PEG-IFN alfa-2a therapy. An 8- to 9-year 

follow-up of the clinical course showed that the HBsAg levels 

decreased at 1.1 Log/mL per year (3.5 Log IU/mL at NA therapy 

initiation Ç 2.4 Log IU/mL at the end of the follow-up) in 

the 33 patients who had received continuous NA therapy and 

at 3.1 Log/mL per year (3.5 Log IU/mL at NA therapy initiation 

Ç 2.5 Log IU/mL at PEG-IFN alfa-2a therapy initiation Ç 

1.7 Log IU/mL at the end of the follow-up period) in those who 

underwent sequential therapy, showing a significant decrease 

(p < 0.0001). [Conclusions] Compared with patients who had 

undergone continuous NA therapy to decrease HBsAg levels, 

those who had received sequential therapy with Peg-IFNα 

-2a after a long-term NA had lower HBsAg levels, suggesting 

sequential therapy may accelerate the timing of HBsAg loss. 

Disclosures: 

The following authors have nothing to disclose: Miwa Kawanaka, Ken Nishino, 

Jun Nakamura, Tomohiro Tanikawa, Takahito Oka, Noriyo Urata, Mitsuhiko 

Suehiro, Hirofumi Kawamoto, Gotaro Yamada


2031 

Impact of pre-treatment platelet count on hepatocellular 

carcinoma development during entecavir treatment of 

chronic hepatitis B virus infection 

Akira Kawano 1 , Hirohisa Shigematsu 1 , Koichiro Miki 1 , Hironori 

Tanimoto 2 , Yasunori Ichiki 10 , Chie Morita 3 , Kimihiko Yanagita 4 , 

Kazuhiro Takahashi 5 , Kazufumi Dohmen 6 , Masataka Seike 7 , 

Hideyuki Nomura 2 , Hiromi Ishibashi 8 , Shinji Shimoda 9 ; 1 Department 

of Medicine, Kitakyushu Municipal Medical Center, Kitakyushu, 

Japan; 2 The Center for Liver Disease, Shin-Kokura Hospital, 

Kitakyushu, Japan; 3 Department of Internal Medicine, JR Kyushu 

Hospital, Kitakyushu, Japan; 4 Department of Internal Medicine, 

Saiseikai Karatsu Hospital, Saga, Japan; 5 Department of Medicine, 

Hamanomachi Hospital, Fukuoka, Japan; 6 Department of 

Internal Medicine, Chihaya Hospital, Fukuoka, Japan; 7 Department 

of Gastroenterology, Oita University, Oita, Japan; 8 Department 

of Hepato-Biliary-Pancreatic Medicine, Fukuoka Sanno Hospital, 

Fukuoka, Japan; 9 Department of Medicine and Biosystemic Science, 

Kyushu University, Fukuoka, Japan; 10 Department of Internal 

Medicine, Japan Community Health care Organization Kyushu 

Hospital, Kitakyushu, Japan 

Background and Aims: Entecavir (ETV) is one of the first-line 

nucleoside analogs for treating patients with chronic HBV infection. 

The risk of the hepatocellular carcinoma (HCC) development 

for ETV-treated patients remains unclear. The objective of 

this study was to evaluate the pre-treatment predictors of the 

development of HCC in the patients of chronic HBV infection 

treated with ETV, and its influence on clinical course. Methods: 

This retrospective multicenter study consisted of 151 Japanese 

patients with chronic HBV infection who received ETV treatment 

more than 12 months. The median age was 51.0 yo and 97 

patients (64.2%) were male. Genotype C was the most prevalent 

(86.0%). Median HBV DNA was 7.1 log copies (LC)/mL. 

Seventy two patients (47.7%) were positive for hepatitis B e 

antigen (HBeAg). Median aminotransferase (ALT) level, serum 

albumin level, platelet (PLT) count and serum alpha-fetoprotein 

(AFP) level were 111 IU/L, 41 g/L, 157 x10 9 /L and 6.6 ng/ 

mL, respectively. Median treatment duration was 47.4 months. 

Results: Fourteen patients developed HCC during follow-up. 

The cumulative incidence of HCC at 3, 5, and 7-year was 7.7, 

11.4, and 13.3 %, respectively. Multivariate analysis extracted 

lower PLT counts (p=0.0055) and higher age (p=0.0133) as 

significant independent pre-treatment predictors of the development 

of HCC. Cumulative incidence of HCC was significantly 

higher in the low PLT group (


antiHBs at the time of NUCs discontinuation. No HBsAg seroreversion 

occurred during a median of 12 months follow-up 

(range, 2-55 months). No patient developed hepatic complications 

or hepatocellular carcinoma. A new 24 months-follow-up 

analysis is currently on-going to evaluate previous outcomes. 

Those results will be presented during AASLD congress. Conclusions: 

These data suggest the time between the diagnosis 

of CHB and HBsAg loss is longer in HBeAg negative than 

in HBeAg positive patients. In addition, HBsAg loss occurred 

faster with TDF/ETV than with LAM/ADV. HBsAg loss was 

maintained in all patients after discontinuation of NUCs. 

Disclosures: 

Emilio Suarez - Advisory Committees or Review Panels: Gilead; Speaking and 

Teaching: Gilead, Bristol Myers Squibb 



Janssen, BMS 


Rafael Gómez Rodríguez - Consulting: Gilead 

Moisés Diago - Grant/Research Support: MSD, JANSSEN; Speaking and Teaching: 

BMS, ABBVIE, GILEAD 

Rosa María Morillas - Advisory Committees or Review Panels: BRISTOL, GILEAD, 

Abvvie; Speaking and Teaching: ROCHE, JANSSEN, MSD 

The following authors have nothing to disclose: Miguel A. Simon, Juan Manuel 

Pascasio, Manuel Rodriguez, Teresa Casanovas, Javier Crespo, Juan I. Arenas, 

Blanca Figueruela, Jose M. Zozaya, José Luís Calleja, Marta Casado, Esther 

Molina, Javier Fuentes 

2033 

Prolongation of interferon therapy increases maintained 

virological response rates and improves the natural 

course of chronic delta hepatitis 

Onur Keskin, Ali Tuzun, Fatih Karakaya, Cagdas Kalkan, Aysun 

Caliskan Kartal, Ersin Karatayli, Senem C. Karatayli, A.Mithat 

Bozdayi, Ramazan Idilman, Cihan Yurdaydin; Gastroenterology, 

Ankara University School of Medicine, Ankara, Turkey 

Introduction and Aim: Chronic delta hepatitis (CDH) is the 

most severe form of chronic viral hepatitis. Interferons are the 

only approved therapy in CDH. HDV RNA 6 months post-treatment 

is an unreliable surrogate of treatment efficacy. Hence 

maintained virologic response (MVR) is important. Aim of 

this study was to assess effect of prolonged interferon therapy 

on MVR and on natural course of CDH. Patients and 

Methods: We searched our CDH database for patients who 

received interferon treatment. 99 CDH patients (70M/29F; 

mean age:49.8±11.3 years, 21 cirrhotic/78 non-cirrhotic at 

baseline) who received at least 6 months of conventional or 

pegylated interferon were identified and included in the analysis. 

MVR was defined as durable negative HDV RNA after 

2 years of post-treatment follow-up. HDV RNA was assessed 

by in-house PCR. Median treatment duration was 24 months 

(6-126) and median number of treatment episodes was 2 (1-8). 

Duration of IFN therapies were classified as 6-12 (n:32),13- 

24 (n:26), 25-36 (n:11), 37-48 (n:15), 49-60 (n:7) and >60 

months (n:8), respectively. Median follow up time after treatment 

discontinuation was 118 (24-255) months. Overall, 35 

patients had a MVR and 64 did not have MVR . Development 

of hepatocellular cancer (HCC), hepatic decompansation, liver 

related mortality, liver transplantation and HBsAg loss were 

considered as clinical end-points. Results: Only 16 patients 

(16%) had MVR with 12 months of IFN therapy. With prolongation 

of IFN treatment, 19 new patients (6, 2, 4, 3 and 4 

patients after 24, 36, 48, 60 and > 60 months of IFN treatment, 

respectively) achieved MVR. Cumulative probability of 

MVR was 16%, 24%, 26%, 30%, 33% and 38% at end of 

6-12, 24, 36, 48, and > 60 months of interferon treatment, 

respectively. Development of HCC (p: 0.05), decompansation 

(p:0.013), liver-related mortality (p:0.03) and transplantation 

(p:0.012) were more common in non-responders and HBsAg 

clearance occurred more often in patients with MVR (p:0.002). 

By multivariate analysis, low platelet count and no response 

to therapy were independent factors for development of any 

clinical endpoint (p=0.014 and 0.021, respectively). Conclusion: 

Longer duration of IFN therapy increases MVR rates (from 

16.1% to 38%) in CDH. Low platelet count at baseline and 

no response to treatment were independent predictors for the 

development of a clinical endpoint. These results suggest that 

treatment with pegylated interferon should be started early in 

CDH and that prolonged periods of treatment may be needed. 

Further, the results underline the urgent need for new drug 

development in CDH. 

Disclosures: 



The following authors have nothing to disclose: Onur Keskin, Ali Tuzun, Fatih 

Karakaya, Cagdas Kalkan, Aysun Caliskan Kartal, Ersin Karatayli, Senem C. 

Karatayli, A.Mithat Bozdayi, Ramazan Idilman 

2034 

TKM-HBV, a Novel RNA Interference Treatment for 

Chronic Hepatitis B, has a Complementary Mode of 

Action to Current Standard of Care Nucleos(t)ide Analogs 

Emily P. Thi, Ammen P. Dhillon, Amy C. Lee; Tekmira Pharmaceuticals, 

Burnaby, BC, Canada 

Current approved nucleos(t)ide analog (NA) drugs for hepatitis 

B virus (HBV) are highly effective at inhibiting viral replication 

but do not cure the chronic infection and are largely ineffective 

in preventing viral protein production. HBV proteins are understood 

to play a variety of roles in contributing to suppression 

of host immune responses and viral persistence. Quantitative 

changes in peripheral HBV surface antigen (HBsAg) in particular 

have been correlated with differences in clinical outcomes. 

TKM-HBV is a novel RNA interference HBV therapeutic 

intervention currently in Phase 1 clinical development. It is a 

mixture of three different oligonucleotide duplexes, encapsulated 

within a lipid nanoparticle delivery system, that cleaves 

all forms of HBV RNAs thus preventing the synthesis of viral 

proteins. Because TKM-HBV is understood to act on a separate 

viral target and in a different cellular compartment than that 

of NAs (in the cytoplasm versus the viral capsid), there is little 

basis to expect drug:drug interference in a co-treatment setting. 

TKM-HBV combination treatment with NA drugs (including the 

current frontline compounds, entecavir and tenofovir) was studied 

in two HBV infection models: primary human hepatocytes 

(PHH) and humanized mice. To mimic a potential clinical study 

setting, TKM-HBV was administered after onset of NA treatment. 

Reductions in HBV DNA and HBsAg were observed in 

PHH cultures co-treated with TKM-HBV and either entecavir, 

tenofovir or lamivudine. No antagonism of either drug activity 

was detected and most often additive effects were observed. 

A second PHH study examining performance against different 

viral genotypes also did not detect any drug:drug interference 

in the ability of entecavir to inhibit viral replication or TKM-HBV 

to reduce viral antigen load when applied against gt A, B, C 

or D. Interestingly, the magnitude of TKM-HBV activity was 

more consistent than that of entecavir across the four genotypes 

examined. The combination approach was also demonstrated 

to be effective in the uPA/SCID humanized mouse model of 

chronic HBV, resulting in simultaneous control of both HBV 

DNA titre and peripheral HBsAg (3 log and 1 log reductions, 

respectively). In summary, these results show that TKM-HBV 

and NA modes of action are complementary, and combination


therapy allows effective disease targeting at multiple critical 

nodes of the viral life cycle. 

Disclosures: 



The following authors have nothing to disclose: Ammen P. Dhillon 

2035 

The New Cyclophilin Inhibitor STG-175 Efficiently Inhibits 

Mono- as well as Co-Infections of HIV-1, HCV and 

HBV 

Philippe Gallay 1 , Michael Bobardt 1 , Udayan Chatterji 1 , Zhengyu 

Long 2 , Shengli Zhang 2 , Zhuang Su 2 ; 1 The Scripps Research Institute, 

La Jolla, CA; 2 S & T Global, Inc., Woburn, MA 

BACKGROUND: Human immunodeficiency virus type-1 (HIV- 

1), hepatitis B virus (HBV) and hepatitis C virus (HCV) share 

a common route of transmission. HBV and HCV mono-infection 

represent the major causes of chronic liver disease globally. 

HIV-1 co-infection with HBV or HCV is associated with 

accelerated progression to severe liver disease, increased risk 

of hepatotoxicity from antiretroviral therapy and reduced survival. 

Co-infected patients are often refractory to most therapies 

and develop liver fibrosis, cirrhosis and liver cancer more 

often than mono-infected patients. Since a growing body of 

evidence suggests that HIV-1, HCV and HBV, all exploit the 

host protein cyclophilin A (CypA) to optimally infect and replicate 

in human cells, we tested a new cyclophilin inhibitor 

STG-175 for its capacity to inhibit mono- as well as co-infections 

of these three prime viral human threats. MATERIAL AND 

METHODS: For mono-infections: i) human PBMCs were infected 

with HIV-1 (JR-CSF) and viral replication was quantified by 

HIV-1 capsid/p24 ELISA; ii) hepatoma Huh7.5.1 cells were 

infected with HCV (JFH-1) and viral replication was quantified 

by HCV core ELISA; and iii) NTCP-positive Huh7 cells were 

infected with HBV AD38 and viral replication was quantified 

by HBV HBeAg ELISA. For dual and triple infections: target 

cell populations were mixed prior to virus exposure. For drug 

treatments, STG-175 was added to cells either i) together with 

viruses or ii) 3 days post-infection and then every 3 days for 

a period of 12 days. RESULTS: In this in vitro co-culture infection 

system, we found that STG-175 inhibits in a dose-dependent 

manner mono-infections, dual co-infections as well as the 

triple HIV-1/HCV/HBV co-infection. The degree of STG-175 

antiviral efficacy was HCV > HIV-1 > HBV. The addition of 

STG-175 together with virus totally blocked HCV and HIV-1 

infection and greatly attenuated HBV infection. When added 

3 days post-infection and then every 3 days, STG-175 totally 

eradicated the pre-established HCV infection, almost totally 

aborted the established HIV-1 infection and prevented the viral 

expansion of the pre-established HBV infection. Similar results 

were observed during the triple HIV-1/HCV/HBV co-infection. 

STG-175 was found to be constantly more efficacious than 

the well-characterized cyclophilin inhibitor alisporivir in both 

mono- and co-infections. CONCLUSIONS: By demonstrating a 

potent and broad spectrum of antiviral activity, the new cyclophilin 

inhibitor STG-175 represents an attractive drug partner 

for an IFN-free regimen for the treatment of HIV-1/HCV/HBV 

co-infections. 

Disclosures: 

The following authors have nothing to disclose: Philippe Gallay, Michael Bobardt, 

Udayan Chatterji, Zhengyu Long, Shengli Zhang, Zhuang Su 

2036 

Association of Interferon-gamma Inducible Protein 10 

Polymorphism with Treatment Response to Pegylated 

Interferon in HBeAg-Positive Chronic Hepatitis B 

Pisit Tangkijvanich 1 , Umaporn Limothai 1 , Natthaya Chuaypen 1 , 

Apichaya Khlaiphuengsin 1 , Rujipat Wasitthankasem 2 , Yong Poovorawan 

3 ; 1 Biochemistry, Chulalongkorn University, Bangkok, Thailand; 

2 Chulalongkorn University, Bangkok, Thailand; 3 Pediatrics, 

Chulalongkorn University, Bangkok, Thailand 

Background: Interferon-gamma inducible protein 10 (IP-10) 

plays an important role in the natural history and treatment 

outcome of patients with chronic hepatitis B (CHB). The aim of 

this study was to investigate the association of single nucleotide 

polymorphism (SNP) G-201A in the promoter region of the IP-10 

gene and treatment response to pegylated interferon (PEG-IFN) 

in patients with HBeAg-positive CHB. The potential effect of 

SNP rs12979860 in the interferon lambda-3 (IFNL3) gene was 

also examined. Method: We retrospectively analyzed data of 

Thai patients with HBeAg-positive CHB treated with PEG-IFN 

for 48 weeks. Virological response (VR) was defined as HBeAg 

clearance and HBV DNA < 2,000 IU/mL at 24 weeks post 

treatment. The SNPs G-201A and rs12979860 were identified 

by PCR–RFLP method. HBV genotype was performed by direct 

sequencing. Baseline serum IP-10 levels were measured by a 

commercially available assay. Results: Among 107 patients 

enrolled (72 male, mean age 34.3 years), VR was achieved 

in 45 (42.1%) patients. HBsAg clearance and HBsAg decline 

(< 100 IU/mL) at 24 weeks post treatment were achieved in 

10 (9.3%) and 22 (20.6%) patients, respectively. The distribution 

of HBV genotypes B and C was 12.1% and 81.3%, 

respectively. The overall distribution of GG, GA and AA genotypes 

of G-201A was 76.6%, 19.6% and 3.7%, respectively. 

Patients with GG genotype, compared to those with non-GG 

genotype, achieved significantly higher rated of VR (48.8% vs. 

19.2%; P=0.011), HBsAg decline (25.6% vs. 4.0%, P=0.019), 

and a trend of HBsAg clearance (11.0% vs. 4%, P=0.294). 

Patients with GG-genotype had more pronounced decline of 

serum HBsAg during and after therapy (weeks 0, 4, 12, 24, 

48 and 72) and had higher baseline IP-10 levels than those 

with non-GG genotype (432.2±339.0 vs. 257.3±145.7 pg/ 

mL, P=0.028). Patients achieving VR had significantly higher 

pretreatment IP-10 levels than non-responders (496.0±394.6 

vs. 328.0±232.2, P=0.013). The distribution of CC, CT and 

TT genotypes of rs12979860 was 89.7%, 10.3% and 0%, 

respectively. Patients with CC and CT genotypes achieved 

VR in 41.7% and 45.5%, respectively (P=0.810). In logistic 

regression analysis, SNP G-201A was an independent factor 

associated with VR (odds ratio 3.81, 95% confidence interval: 

1.31-11.12, P=0.014). Conclusions: These data demonstrated 

that IP-10 polymorphism, but not IFNL3 polymorphism, was 

independently associated with response to PEG-IFN therapy in 

Thai patients with HBeAg-positive CHB. 

Disclosures: 

The following authors have nothing to disclose: Pisit Tangkijvanich, Umaporn Limothai, 

Natthaya Chuaypen, Apichaya Khlaiphuengsin, Rujipat Wasitthankasem, 

Yong Poovorawan


2037 

Whole-Genome Sequencing of Patients Achieving Hepatitis 

B Virus Serum Antigen Loss Following TDF Treatment 

Sarah E. Kleinstein 1,2 , Harry L. Janssen 3,4 , Patrick Marcellin 5,6 , 

Dongliang Ge 7 , Anuj Gaggar 7 , Mani Subramanian 7 , John G. 

McHutchison 7 , Maria Buti 8 , Alex J. Thompson 9 , David B. Goldstein 

1 ; 1 Center for Human Genome Variation, Duke University, 

New York, NY; 2 Molecular Genetics & Microbiology, Duke University, 

Durham, NC; 3 Toronto Centre for Liver Disease, University 

Health Network, Toronto, ON, Canada; 4 Erasmus Medical Center, 

Rotterdam, Netherlands; 5 Viral Hepatitis Research Centre, Clichy, 

France; 6 Service d’Hépatologie, Hôpital Beaujon, Clichy, France; 

7 Gilead Sciences Inc., Foster City, CA; 8 Hospital Valle Hebron and 

Ciberehd del Institut Carlos III, Barcelona, Spain; 9 St Vincent’s Hospital 

and the University of Melbourne, Melbourne, VIC, Australia 

Purpose: While treatment with tenofovir disoproxil fumerate 

(TDF) provides durable suppression of hepatitis B virus (HBV), 

serum antigen (HBsAg) loss, which is considered a “cure”, is 

infrequent; thus, we investigated the role of rare human genetic 

variants in predicting HBsAg loss following treatment with TDF 

among chronic hepatitis B (CHB) patients. Methods: 10 Caucasian 

patients with CHB (HBeAg+) achieved HBsAg loss following 

treatment in a Phase 3 study, consented, and had available 

DNA. Whole-genome sequencing (WGS) was performed on 

patient DNA using the Illumina HiSeq2000 (v3 chemistry). 

The genetic data from the 10 HBsAg loss cases were compared 

to 122 healthy Caucasian WGS controls, matched on 

sequencing platform and chemistry. All samples underwent 

standard quality control checks. Case-control gene-based and 

single-variant (Fisher’s exact test) associations were run across 

all genetic models. Gene-based analyses were limited to coding 

region, functional variants. A Bonferroni-corrected p-value 

for the number of genes or variants tested was set as the statistical 

significance threshold. Results: WGS analysis of the 10 

HBsAg loss patients did not identify any rare variants that conferred 

a significant beneficial response to TDF when analyzed 

at either the gene or single-variant level. However, non-significant 

suggestive associations were identified in 9 genes. 

Rare coding variants enriched among patients with HBsAg loss 

were identified in: PDCD1, IL2RA, SCTR, IFI16, ULK4, GPA33, 

CCHCR1, GMEB1, and GPNMB. PDCD1 has been associated 

with impaired immune response in CHB. SCTR, GPA33 and 

CCHCR1 have been linked to the natural history of CHB. Variants 

were identified under a dominant (gene-based) or allelic 

(single-variant) model, except for IFI16 (recessive model) and 

ULK4 (compound-heterozygous model). Conclusions: WGS of 

a small cohort of Caucasian CHB patients with HBsAg loss 

following TDF treatment identified enrichment of several interesting 

rare variants. Variants were identified in genes involved 

in the immune system and/or virus replication. It is currently 

unclear whether these identified genes and variants play a role 

in HBsAg loss or control of HBV replication. Validation studies 

are planned in an independent cohort. 

Rare coding variants enriched among patients with HBsAg loss 

a Gene-based; 

b Different control variant 

Disclosures: 










Abbvie 





Sciences 



Janssen, BMS 




Abbvie, BMS 

David B. Goldstein - Advisory Committees or Review Panels: Sever Adverse 

Events Consortium; Grant/Research Support: Gilead, Astra Zeneca, Biogen, 

Johnson and Johnson, Labcorp, Merck, Fundazione Telethon, NIH, CURE, UCB, 

IL28B and ITPA finds 

The following authors have nothing to disclose: Sarah E. Kleinstein


2038 

A randomized trial evaluating the antiviral efficacy of 

switching from Lamivudine plus Adefovir to Tenofovir 

disoproxil fumarate monotherapy in Lamivudine-resistant 

chronic hepatitis B patients with undetectable hepatitis 

B virus DNA - The 48 week interim analysis 

Heon Ju Lee 2 , Jeong Min Kim 1 , Young Oh Kweon 3 , Soo Young 

Park 3 , Jeong Heo 4 , Hyun Young Woo 4 , Jaeseok Hwang 1 , Woo 

Jin Chung 1 , Chang Hyeong Lee 5 , Byung Seok Kim 5 , Jeong Ill Suh 6 , 

Won Young Tak 3 , Byoung Kuk Jang 1 ; 1 Internal medicine, Keimyung 

University School of Medicine, Daegu, Korea (the Republic of); 

2 Internal medicine, Yeungnam University College of Medicine, 

Daegu, Korea (the Republic of); 3 Internal medicine, Kyungpook 

National University College of Medicine, Daegu, Korea (the 

Republic of); 4 Internal medicine, Pusan National University School 

of Medicine, Busan, Korea (the Republic of); 5 Internal medicine, 

Catholic University of Daegu College of Medicine, Daegu, Korea 

(the Republic of); 6 Internal medicine, Dongguk University College 

of Medicine, Gyeongju, Korea (the Republic of) 

Introduction: Tenofovir disoproxil fumarate (TDF) monotherapy 

is an effective treatment for patients who have lamivudine 

(LAM)-resistant chronic hepatitis B (CHB). However, the efficacy 

of switching to TDF monotherapy for LAM resistant CHB patient 

with undetectable HBV DNA while on LAM plus adefovir (ADV) 

combination therapy (stable switching) is not well established. 

Material and Methods: In this non-inferiority trial, LAM-resistant 

CHB patients who had undetectable serum HBV DNA (40 IU/mL at two consecutive timepoints, 

or persistent HBV DNA levels of 20-40 IU/mL at three 

consecutive timepoints. Results: A total of 169 CHB patients 

were enrolled in this study including 74(43.8%) with compensated 

cirrhosis. There were no significant differences between 

two groups in age, gender, biochemistry findings and duration 

of LAM and ADV combination therapy at baseline. Twelve 

(20.7%) patients in the LAM/ADV group and 18 (16.2%) 

patients in TDF group were HBeAg-positive. Nine patients (4 in 

the LAM/ADV group and 5 in the TDF group) discontinued the 

study. After a mean follow up period of 48 weeks, there were 

no subjects in either group who experienced viral reactivation. 

The number of patients with HBeAg loss at week 48 was 

2/12(16.7%) and 3/18(16.7%) in the LAM/ADV and TDF 

groups, respectively. Two patients achieved HBeAg seroconversion 

(1/12 [8.3%] in LAM/ADV group and 1/18 [5.6%] 

in TDF group). Transient virological rebound occurred in 10 

patients (5 patients in LAM/ADV group and 5 patients in TDF 

group) through 48 weeks; however, all patients subsequently 

achieved HBV DNA undetectablility at the next study visit. No 

patient experienced significant increases in serum creatinine 

levels above baseline. The mean changes in serum creatinine 

levels of both groups were minimal throughout the 48 weeks of 

treatment. Conclusions: Stable switching to TDF monotherapy 

for 48 weeks demonstrated a comparable virological response 

to maintaining LAM plus ADV combination therapy in LAM-resistant 

CHB patients with undetectable HBV DNA. (Trial registration 

number Clinicaltrial.gov ID NCT01732367) 

Disclosures: 

Jeong Heo - Grant/Research Support: Roche 



The following authors have nothing to disclose: Heon Ju Lee, Jeong Min Kim, 

Young Oh Kweon, Soo Young Park, Hyun Young Woo, Jaeseok Hwang, Woo 

Jin Chung, Chang Hyeong Lee, Byung Seok Kim, Jeong Ill Suh, Byoung Kuk Jang 

2039 

WITHDRAWN 

2040 

Association between Pre-S2 Deletions and Virologic 

Rebound during Peginterferon Therapy in Patients with 

Hepatitis B E Antigen-Positive Chronic Hepatitis B 

Cheng-Yuan Peng 1 , Chao-Jen Shih 2 , Pei-Shuan Lo 1 , Chia-Lin 

Huang 1 , Chia-Hsin Lin 1 , Hsueh-Chou Lai 1 , Wen-Pang Su 1 ; 1 Division 

of Hepatogastroenterology, Department of Internal Medicine, 

China Medical University Hospital, Taichung, Taiwan; 2 Agricultural 

Biotechnology Research Center, Academia Sinica, Taipei, 

Taiwan 

Background: To determine the incidence of on-treatment virologic 

rebound (VR) during peginterferon (Peg-IFN) therapy in 

patients with HBeAg-positive chronic hepatitis B (CHB) and its 

association with genomic mutations in hepatitis B virus (HBV). 

Methods: We analyzed the on-treatment HBV DNA kinetics 

in 103 consecutive HBeAg-positive CHB patients treated with 

Peg-IFN for 24 (n=58) or 48 weeks (n=45). On-treatment 

serum HBV DNA levels were measured every 3 months (Cobas 

Amplicor HBV Monitor Test, Roche Diagnostics, sensitivity: 

60 IU/mL). VR was defined as an on-treatment increase in 

HBV DNA level of >1 log IU/mL over prior nadir level with or 

without an initial decline. Full-length HBV genomic population 

sequencing was done at baseline, nadir, VR, and 24 weeks 

off therapy. Precore and basal core promoter mutations and 

the pre-S2 deletions were examined at baseline. Proportions 

of the pre-S2 deletion mutants among total viral populations 

were measured using quantitative PCR at baseline, nadir, VR, 

and 24 weeks off therapy. The replication capability of HBV 

mutants was determined after transient transfection with the 

pTriEx-HBV vector in Huh7 cell line. Results: Twelve (12%) 

patients displayed on-treatment VR. Univariate analysis identified 

HBV DNA


Disclosures: 

Cheng-Yuan Peng - Advisory Committees or Review Panels: AbbVie, BMS, Gilead, 

MSD, Roche 

The following authors have nothing to disclose: Chao-Jen Shih, Pei-Shuan Lo, 

Chia-Lin Huang, Chia-Hsin Lin, Hsueh-Chou Lai, Wen-Pang Su 

2041 

WITHDRAWN 

2042 

Is efficacy of tenofovir in nucleos(t)ide analogue-experienced 

chronic hepatitis B patients comparable to that in 

nucleos(t)ide analogue-naïve patients? 

Eun Ju Cho 1 , Jeong-Hoon Lee 1 , Jeong-Ju Yoo 1 , Yuri Cho 1 , Donghyeon 

Lee 1 , Yun Bin Lee 2 , Su Jong Yu 1 , Yoon Jun Kim 1 , Jung-Hwan 

Yoon 1 ; 1 Internal Medicine, Seoul National University Hospital, 

Seoul, Korea (the Republic of); 2 Internal Medicine, CHA Bundang 

Medical Center, Seoungnam, Korea (the Republic of) 

Background/Aim: Recent study reported that entecavir fails to 

show comparable efficacy in nucleos(t)ide analogue (NA)-experienced 

chronic hepatitis B (CHB) patients as compared 

to NA-naïve ones. We evaluated the efficacy of tenofovir in 

NA-experienced CHB patients in comparison with NA-naïve 

patients. Methods: We retrospectively included 221 consecutive 

patients who had serum hepatitis B virus (HBV) DNA 

level > 2,000 IU/mL at the initiation of tenofovir treatment 

and received tenofovir for > 3 months. Complete virologic 

response (CVR) was defined as undetectable serum HBV DNA 

by real-time PCR. CVR rates were calculated by Kaplan-Meier 

analysis, and a multivariate Cox proportional-hazard model 

was generated in order to find predictive factors independently 

associated with the time to a CVR. Results: Of the 221 patients 

(129 males; mean age, 48.2 years), 149 were NA-naïve and 

72 were NA-experienced patients. Ninety-eight patients had 

hepatitis B e antigen (HBeAg)-positive CHB. The median duration 

of tenofovir treatment was 47.4 (interquartile range, 36.2 

to 56.6) weeks. The CVR rates of the NA-naïve and NA-experienced 

groups were comparable in both HBeAg-negative 

(49.4% vs 58.1 at week 24, P=0.53; 96.4% vs 93.3% at 

week 48, P=0.60) and HBeAg-positive patients (22.2% vs 

16.7% at week 24, P=0.60; 75.6% vs 75.9% at week 48, 

P=1.00). According to the multivariate Cox regression model, 

only HBeAg positivity (hazard ratio [HR], 0.40; 95% confidence 

interval [CI], 0.29 to 0.56; P< 0.001) and baseline 

HBV DNA levels (HR, 0.77 for 1 log10 IU/mL increase; 95% 

CI, 0.69 to 0.86; P< 0.001) had a significant influence on the 

time to a CVR. The presence of resistance mutations to lamivudine/telbivudine/entecavir 

did not influence the response 

rates, however, patients with adefovir resistance showed significantly 

lower CVR rate (25%; P=0.03). Conclusion: Tenofovir 

has comparable efficacy in NA-naïve and NA-experienced 

patients without adefovir resistance. 

Disclosures: 






The following authors have nothing to disclose: Eun Ju Cho, Jeong-Hoon Lee, 

Jeong-Ju Yoo, Yuri Cho, Donghyeon Lee, Yun Bin Lee, Su Jong Yu, Jung-Hwan 

Yoon 

2043 

Serologic and Virologic Changes Following ALT flares 

in adolescents With Chronic Hepatitis B during therapy 

with Tenofovir Disoproxil Fumarate or Placebo. 

Karen F. Murray 1 , Leszek Szenborn 2 , Jacek Wysocki 3 , Benedetta 

Massetto 4 , Anuj Gaggar 4 , Kyungpil Kim 4 , Mani Subramanian 4 , 

Luminita Luminos 5 , Malgorzata Pawlowska 6 , Jacek Mizerski 7 ; 

1 Children’s Hospital, Seattle, WA; 2 Wroclaw Medical University, 

Wroclaw, Poland; 3 Poznan University of Medical Sciences, 

Poznan, Poland; 4 Gilead Sciences, Inc., Foster City, CA; 5 Prof. Dr. 

Matei Bals Institute for Infectious Diseases, Bucharest, Romania; 

6 Wojewodzki Szpital Obserwacyjno-Zakazny im. T. Browicza, 

Bydgoszcz, Poland; 7 John Paul II Hospital, Kraków, Poland 

Background and Aims: We previously described the safety and 

antiviral efficacy of TDF in adolescents aged 12 to 2x baseline and 

>5x upper limit of normal during the first 48 weeks. HBeAg 

loss, HBV DNA decline and HBsAg decline were evaluated 24 

weeks following a flare. Results: Twelve of the 52 (23%) evaluable 

PBO patients and 1 of the 52 (2%) TDF-treated patients 

had ALT flares during the first 48 weeks. PBO patients with 

ALT flare had an increased rate of 1-log 10 

HBV DNA decline 

and HBeAg loss compared to those without ALT flare (Table, 

p




The following authors have nothing to disclose: Leszek Szenborn, Kyungpil Kim, 

Luminita Luminos, Malgorzata Pawlowska, Jacek Mizerski 

2044 

The baseline combination of HBcrAg and HBsAg titers 

enhance treatment outcome and HBsAg loss predictions 

in HBeAg negative chronic hepatitis B patients treated 

with pegylated interferon alfa-2a or PegIFN plus Tenofovir-disoproxil-fumarate. 

Michelle Martinot-Peignoux 1,2 , Martine Lapalus 1 , Nathalie Boyer 2 , 

Corinne Castelnau 2 , Nathalie Giuily 2 , Michele Pouteau 2 , Rami 

Moucari 1 , Tarik Asselah 2,1 , Patrick Marcellin 2,1 ; 1 Université Denis 

Diderot Paris 7, INSERM, UMR1149, Centre de Recherche sur 

l’Inflammation, Clichy, France; 2 Hôpital Beaujon AP-HP, Service 

d’Hépatologie, Clichy, France 

Background. It remains unclear whether adding a nucleot(s)ide 

analogue (NAs) enhances the efficacy of pegylated interferon 

alfa-2a (PegIFN) by accelerating HBsAg clearance. Baseline 

HBsAg level is predictor of SVR in patients receiving PegIFN. 

Hepatitis B core-related antigen (HBcrAg) is a new serological 

HBV marker that could to be used for NAs treatment cessation. 

It combines 3 viral proteins coded by the precore/core region 

of the covalently closed circular DNA: HBeAg, HBcAg and 

a core related protein (p22cr) by sharing a 149 amino-acid 

sequence. We aimed to evaluate if the baseline combination 

of HBcrAg and HBsAg levels might have a complimentary role 

predicting treatment response, in patients receiving PegIFN) 

therapy or PegIFN plus Tenofovir-disoproxil-fumarate (PegIF- 

N+TDF) Patients-Methods. 62 patients HBeAg negative CHB 

patients were treated with 48 weeks PegIFN or PegIFN+TDF. 

Patients were followed 3 years after treatment cessation. HBsAg 

and HBcrAg titers measured at baseline. HBsAg and HBV DNA 

were measured at baseline, week 24, end of therapy (EOT), 

24 weeks, 1, 2 and 3 years after treatment cessation. Results. 

30 patients received PegIFN and 32 patients received PegIF- 

N+TDF. The 2 groups of patients were similar with regards to 

age, sex ratio, ALT levels, HBV genotypes distribution, histologic 

lesions, mean HBsAg, HBcrAg and HBV DNA titers. An 

EOT response was observed in 90% and 100% of PegIFN and 

PegIFN+TDF patients, respectively. SVR was 10/30 (33%) and 

17/32 (53%) in PegIFN and PegIFN+TDF patients, respectively. 

At the end of 3 years post-treatment follow-up an HBsAg 

loss was observed in 7/30 (1 at EOT) and 6/32 (4 at EOT) in 

PegIFN and PegIFN+TDF patients, respectively. Baseline negative 

predictive values (NPVs) for SVR of HBsAg threshold 3.302 

log IU/ml and HBcrAg threshold 3.699 log U/ml and combination 

of both thresholds were: 78% or 63%, 74% or 61% 

and 80% or 75% in patients receiving PegIFN or PegIFN+TDF, 

respectively. Baseline NPVs for HBsAg loss of HBsAg threshold 

3.302 log IU/ml and HBcrAg threshold 3.699 log U/ml and 

combination of both thresholds were: 88% or 100%, 79% or 

89% and 87% or 100% in patients receiving PegIFN or PegIF- 

N+TDF, respectively. Conclusions. In HBe negative patients, 

our results indicate that the addition of TDF to PegIFN enhance 

the rate of viral suppression with 53% SVR and 12.5% EOT 

HBs loss, while SVR was observed in 33% and EOT HBs loss 

in 3.3% patients receiving with PegIFN monotherapy. Baseline 

combination of HBsAg and HBcrAg titers allows the identification, 

prior therapy, of patients with low probability of SVR and 

HBsAg loss indicating that these markers could be used for “à 

la carte therapy”. 

Disclosures: 









Abbvie 

The following authors have nothing to disclose: Michelle Martinot-Peignoux, 

Martine Lapalus, Corinne Castelnau, Nathalie Giuily, Michele Pouteau, Rami 

Moucari 

2045 

Baseline HBsAg titer allows identification of patients 

that will benefit from pegylated interferon therapy and 

experience an HBsAg loss. 

Michelle Martinot-Peignoux 2,3 , Nathalie Boyer 3 , Corinne Castelnau 

3 , Nathalie Giuily 3 , Michele Pouteau 3 , Martine Lapalus 1,2 , Rami 

Moucari 1,2 , Tarik Asselah 3,1 , Patrick Marcellin 3,2 ; 1 Université Paris 

Diderot, INSERM U773/CRB3 et Service d’hépatologie, Clichy, 

France; 2 Université Denis Diderot Paris 7, INSERM, UMR1149, 

Centre de Recherche sur l’Inflammation, Clichy, France; 3 Hôpital 

Beaujon AP-HP, Service d’Hépatologie, Clichy, France 

Background. A 48 weeks treatment with pegylated interferon 

alfa-2a (PegIFN) is the only treatment allowing to obtain 30% of 

sustained virological response (SVR) and 10% HBs loss within 

3 years post-treatment follow-up. It is well demonstrated that 

early on treatment HBs decline is predictive of response. We 

aimed to evaluate the predictive value of baseline HBsAg titer 

to predict SVR and HBs loss, in patients treated with PegIFN or 

PegIFN plus Tenofovir-disoproxil-fumarate (PegIFN+TDF) Methods. 

62 HBeAg(-) CHB patients treated for 48 weeks. HBsAg 

and HBVDNA titers were measured at baseline, end of therapy 

(EOT), 24 weeks, 1 2 and 3 years after treatment cessation. 

HBV genotype distribution at baseline. Results. 30 patients 

received PegIFN and 32 patients received PegIFN+TDF. An 

EOT response was observed in 90% and 100% of PegIFN and 

PegIFN+TDF patients, respectively. An SVR was observed in 

27/ 62 (43%), 10/30 (33%) and 17/32 (53%) in the overall, 

PegIFN and PegIFN+TDF patients, respectively. At the end of 

3 years post-treatment follow-up an HBsAg loss was observed 

in 13/62 (21%) 7/30 (23%) and 6/32 (19%), in the overall, 

PegIFN and PegIFN+TDF patients, respectively. HBsAg and 

HBV DNA titers were significantly higher in SVR than in non-responders 

(p2000 IU/ml including 

27/35 (77%) non-responders patients and 36/49 (74%) 

patients with no-HBs loss within 3 years post-treatment follow-up. 

The positive predictive value of HBsAg titer ≤ 2000UI/ 

ml for SVR and HBs loss were 67% and 46% or 55% and 

45% or 77% and 46% in the overall population or patients 

receiving PegIFN or PegIFN+TDF, respectively. The negative 

predictive value (NPVs) of baseline HBsAg titer >2000 IU/ml 

for SVR and HBs loss were 71% and 85% or 79% and 89% or 

63% and 100% in the overall population or patients receiving 

PegIFN or PegIFN+TDF, respectively. The NPVs for genotypes 

A, B, D, and E, were 50%, 80%, 74% and 82%, respectively. 

Conclusions. Our results show that at baseline an HBsAg threshold 

of 2000 IU/ml is highly predictive of treatment response 

and HBsAg loss in patients treated with 48 weeks of PegIFN 

patients or PegIFN+TDF. Among the patients with an HBsAg 

level ≤ 2000 IU/ml 59% demonstrated a SVR and 85% expe-


rienced HBs loss within 3 years after treatment cessation. This 

observation demonstrates for the first time that baseline HBsAg 

titer might allows identifying patients that could or could not 

benefit from PegIFN therapy. 

Disclosures: 









Abbvie 

The following authors have nothing to disclose: Michelle Martinot-Peignoux, 

Corinne Castelnau, Nathalie Giuily, Michele Pouteau, Martine Lapalus, Rami 

Moucari 

2046 

Intrahepatic IP-10 expression correlates with plasma 

IP-10 levels and is a response marker for HBeAg-positive, 

but not HBeAg-negative chronic hepatitis B patients 

treated with peginterferon and adefovir 

Sophie Willemse 2 , Louis Jansen 2 , Annikki de Niet 2 , Marjan J. Sinnige 

1 , Bart Takkenberg 2 , Joanne Verheij 3 ; 1 Experimental Immunology, 

Academic Medical Center, Amsterdam, Netherlands; 

2 Gastroenterology and Hepatology, Academic Medical Center, 

Amsterdam, Netherlands; 3 Pathology, Academic Medical Center, 

Amsterdam, Netherlands 

Background and aims Interferon-y–inducible protein-10 (IP-10) 

is an interferon-stimulated gene that is produced by different 

types of cells such as monocytes, neutrophils and hepatocytes. 

After binding to its receptor CXCR3, IP-10 functions as a chemotactic 

cytokine for T-lymphocytes, monocytes and NK-cells 

and induces adhesion of activated memory/effector T-cells. 

We aimed to establish if IP-10 expression in liver tissue and 

IP-10 levels in plasma of chronic hepatitis B (CHB) patients 

correlated with each other and further to investigate if IP-10 

levels could predict treatment outcome in CHB patients treated 

with peginterferon and adefovir. Methods A total of 86 CHB 

patients (41 HBeAg-positive and 45 HBeAg-negative) received 

combination therapy of peginterferon and adefovir for 48 

weeks. Combined Response (CR) (HBeAg-negativity, HBV-DNA 

≤2,000 IU/mL, and ALT normalization) and non-response (NR) 

were assessed at Week 72. Plasma IP-10 levels were measured 

at baseline and during treatment at Day 3 (D3) and 

Week 1 (W1). Pre-treatment liver biopsies were obtained in 

69 of 86 patients, of which 41 were stored in liquid nitrogen, 

enabling the analysis of intrahepatic IP-10 expression by 

RT-qPCR. Results CR was achieved in 14/41 HBeAg-positive 

and 17/45 HBeAg-negative patients. Mean baseline plasma 

IP-10 levels were significantly higher in HBeAg-positive patients 

with CR than NR (3.20 vs 3.00 log pg/mL p=0.023). This 

difference was not observed in HBeAg-negative patients. Baseline 

IP-10 levels correlated with ALT-levels in HBeAg-positive 

(r0.66 p


Disclosures: 

Nowlan Selvapatt - Speaking and Teaching: Gilead, BMS 

Ashley S. Brown - Advisory Committees or Review Panels: MSD, Roche, Bristol-Myers-Squibb, 

Gilead, Janssen, Abbvie, Achillion; Speaking and Teaching: 

MSD, Roche, Bristol-Myers Squibb, Gilead, Janssen, Abbvie 

The following authors have nothing to disclose: Bilal Khan, Abbesega Ananthavarathan 

2048 

A Multicenter, Prospective, Observational, Non-interventional 

Cohort Study in Chinese Subjects with HBeAg 

Negative Chronic Hepatitis B Receiving Therapy with 

Peginterferon alfa: An Interim Analysis 

Xinyue Chen 1 , Qianguo Mao 2 , Hui Li 3 , Xu Li 4 , Yuqin Xu 5 , Yao 

Xie 6 , Yingxia Liu 7 , Weiping Zhu 8 , Xiaoguang Dou 9 , Fusheng 

Zhu 10 , Huanwei Zheng 11 , Shuqin Zhang 12 , Mingxiang Zhang 13 , 

Yongfeng Yang 14 , Yaoren Hu 15 , Chuanwu Zhu 16 , Jiguang Ding 17 , 

Youwen Tan 18 , Qing Xie 21 , Jifang Sheng 19 , Zhiliang Gao 20 , Yi 

Xie 21 , Hong Ren 22 , Jidong Jia 23 ; 1 Beijing YouAn Hospital, Capital 

Medical University, Beijing, China; 2 Xiamen Hospital of Traditional 

Chinese Medicine, Xiamen, China; 3 The Third People’s 

Hospital of KunMing City, Kunming, China; 4 The First Affiliated 

Hospital of Anhui Medical University, Hefei, China; 5 The 211 

Hospital of People’s Liberation Army, Haerbin, China; 6 Beijing 

Ditan Hospital, Capital Medical University, Beijing, China; 7 Shenzhen 

Third People’s Hospital, Shenzhen, China; 8 QingDao Infectious 

Diseases Hospital, Qingdao, China; 9 Shengjing Hospital of 

China Medical University, Shenyang, China; 10 General Hospital 

of Dagang Oilfield, Tianjin, China; 11 The Fifth Hospital of Shijiazhuang, 

Shijiazhuang, China; 12 Hepatology Hospital of Jilin 

Province, Changchun, China; 13 The Sixth People’s Hospital of 

Shenyang, Shenyang, China; 14 The Second Hospital of Nanjing, 

Nanjing, China; 15 Ningbo No. 2 Hospital, Ningbo, China; 16 The 

Fifth People’s Hospital of Suzhou, Suzhou, China; 17 Ruian People’s 

Hospital, Ruian, China; 18 The Third People’s hospital of Zhenjiang, 

Zhenjiang, China; 19 The First Affiliated Hospital of Zhejing 

University, Hangzhou, China; 20 The Third Affiliated Hospital, Sun 

Yat-sen University, Guangzhou, China; 21 Shanghai Roche Pharmaceuticals 

Ltds., Shanghai, China; 22 The Second Affiliated Hospital 

of Chongqing Medical University, Chongqing, China; 23 Beijing 

Friendship Hospital, Capital Medical University, Beijing, China 

Objectives: To evaluate the efficacy and safety of Peginterferon 

alfa (Peg-IFNα) in Chinese patients with HBeAg negative 

chronic hepatitis B (CHB) in routine clinical practice. Methods: 

Chinese adult HBeAg negative CHB patients, not co-infected 

with HIV, HAV, or HCV, not on concomitant telbivudine therapy, 

and treated with Peg-IFNα (dosing in accordance with 

approved SmPC in China and as per Chinese standard clinical 

practice) were included in a prospective, multicenter, observational, 

non-interventional cohort study. An interim analysis 

of efficacy and safety of Peg-IFNα was performed for the first 

50% patients at 48 weeks on-treatment (end of standard treatment). 

Results: The study included 503 patients (80.7% of male) 

with mean age of 37.5 years, 2.84 log IU/mL mean baseline 

HBsAg and 5.27 log IU/mL mean baseline HBV-DNA. Of 

these, 411 patients (81.7%) completed the 48-week treatment. 

Response rate of HBV-DNA suppression (HBV-DNA


CLUSION: The baseline HBV DNA level and the duration of 

consolidation therapy in inactive carriers were indicators for 

SOVR after stopping preemptive AVT. 

Disclosures: 

The following authors have nothing to disclose: Seung Hyeon Jang, Hyo Jin Kim, 

jung hee kim, Dong Hyun Sinn, Geum-Youn Gwak, Yong-Han Paik, Moon Seok 

Choi, Joon Hyeok Lee, Kwang Cheol Koh, Seung Woon Paik, Byung Chul Yoo 

2050 

Prediction of sustained off-treatment response in chronic 

hepatitis B patients undergoing pegylated interferon 

treatment by a new scoring model 

Yin-Chen Wang 1 , Yi-Hsiang Huang 1,3 , Sien-Sing Yang 2 , Chien- 

Wei Su 1 , Kuei-Chuan Lee 1 , Han-Chieh Lin 1 ; 1 Taipei Veterans 

General Hospital, Division of Gastroenterology, Department of 

Medicine, Taipei City, Taiwan; 2 Cathay General Hospital Medical 

Center, Liver Center, Taipei, Taiwan; 3 National Yang-Ming University, 

Institute of Clinical Medicine, Taipei, Taiwan 

Background The pegylated interferon-α-2a (PEG-IFN-α2a) is 

one of the first line treatment options for chronic hepatitis B 

(CHB). However, the overall response rate is not satisfactory 

mainly due to the lack of a practical model to select patients 

who would have the best chance of response. The aim of this 

study was to develop a new simple scoring model to predict sustained 

off-treatment response of PEG-IFN-α2a in CHB patients. 

Methods We retrospectively reviewed consecutive 201 CHB 

patients who underwent PEG-IFN-α2a treatment from Taipei 

Veterans General Hospital and Cathay General Hospital. Of 

them, 118 were hepatitis B e antigen (HBeAg)-positive and 83 

were HBeAg-negative. Sustained off-treatment response was 

defined as HBeAg seroconversion in accompanied with HBV 

DNA less than 2,000 IU/ml at 48 weeks post treatment for 

HBeAg-positive patients, and HBV DNA less than 2,000 IU/ml 

at 48 weeks post treatment for HBeAg-positive patients. Multivariate 

Logistic regression was performed to determine factors 

associated with the response. Results For HBeAg-positive CHB 

patients, baseline hepatitis B s antigen (HBsAg) 5 ULN CK elevation and progressive 

muscular symptoms, it was planned to evaluate the course of 

treatment to stop or to go on. Results Total 178 patients (105 

male, 73 female) were included to the study. Mean age was 

39.9±11.4 year (17-72 years) and mean duration of therapy 

25 months (6-40 months). Mean serum CK level at the admission 

was 133±106 U/L (34-823 U/L). In 66 patients (37%) 

CK level was remained normal throughout the treatment. The 

highest level of CK was 2221 U/L during the treatment. Significant 

elevation of CK levels were observed after ninth month of 

therapy (p


2052 

Biologic Responses to GS-4774 in Virally-Suppressed 

Chronic HBV Patients 

Carlo Ferrari 1 , Edward J. Gane 2 , Steven-Huy B. Han 3 , W. Jeffrey 

Fessel 4 , Huy N. Trinh 5 , Tim Rodell 6 , Anuj Gaggar 7 , Benedetta 

Massetto 7 , Ondrej Podlaha 7 , Ann D. Johnson 7 , Leanne Peiser 7 , 

Jacky Woo 7 , Mani Subramanian 7 , John G. McHutchison 7 , Stuart 

C. Gordon 8 , Calvin Q. Pan 9 , Hannah Lee 10 , Anna S. Lok 11 ; 

1 University of Parma, Parma, Italy; 2 Auckland General Hospital, 

Auckland, New Zealand; 3 Ronald Reagan UCLA Medical Center, 

Los Angeles, CA; 4 Kaiser Permanente, San Francisco, CA; 5 San 

Jose Gastroenterology, San Jose, CA; 6 Globeimmune, Louisville, 

CO; 7 Gilead Sciences, Inc., Foster City, CA; 8 Henry Ford Hospital, 

Detroit, MI; 9 Medical Procare, PLLC, Flushing, NY; 10 Tufts Medical 

Center, Boston, MA; 11 University of Michigan Health System, Ann 

Arbor, MI 

Background: Chronic HBV (CHB) infection is characterized by 

a dysfunctional HBV-specific T-cell response that permits the 

persistence of infected hepatocytes. GS-4774 is a heat-inactivated 

yeast-based therapeutic T-cell vaccine that expresses a 

recombinant protein comprised of the HBV X, surface, and core 

proteins designed to elicit an HBV-specific T-cell response. This 

study evaluated GS-4774 in virally-suppressed CHB patients. 

Methodology: 178 patients with CHB were randomized to 

continue oral antiviral treatment (OAV) alone or to continue 

OAV and receive GS-4774 (2, 10 or 40 YU; 1YU=10^7 yeast 

cells) every 4 weeks for 6 doses. At Week 48 ex vivo IFN-γ 

ELISpot was done using PBMCs and recombinant proteins and 

overlapping 15 mer peptides spanning the HBV core, S and X 

regions in GS-4774. ELISpot response was quantified as total 

spots/10^6 cells. Flow cytometry was performed at baseline, 

Weeks 12, 24, and 48 using whole blood. RNASeq from 

whole blood was performed on a subset of patients (N=30) at 

baseline and at Week 24. Results: ELISpot results were available 

for 108 patients. The strength of response, as determined 

by absolute IFN-γ spot counts to HBV protein and peptides, was 

generally higher among GS-4774 treated patients particularly 

among the HBeAg- patients, compared to controls. These differences 

were most consistent with HBcAg proteins and peptides 

and was significant among HBeAg- patients (p


levels increased significantly during therapy (8.1 vs. 11.9 vs. 

15.5ng/ml, p


2056 

Long-term Renal Safety of Adefovir Dipivoxil Treatment 

: Focused on development of Fanconi Syndrome 

Sung Won Cho, Soo Yeon Cho, Choong Kyun Noh, Soon Sun 

Kim, Dukyong Yoon, Rae Woong Park, In-Whee Park; Ajou University 

School of Medicine, Suwon, Korea (the Republic of) 

Background Adefovir dipivoxil (ADV) is a commonly used 

antiviral agent for the treatment of hepatitis B virus or human 

immunodeficiency virus infection. ADV exhibits dose-dependent 

nephrotoxicity that can cause Fanconi syndrome. Fanconi 

syndrome is a generalized dysfunction of the proximal renal 

tubule resulting in impaired reabsorption and increased urinary 

loss of phosphate and other solutes such as uric acid, glucose, 

amino acids and bicarbonate. Although low-dose ADV therapy 

(10mg/day) has been reported to be safe, there are increasing 

number of reports stating that the long-term use of lowdose 

ADV causes Fanconi syndrome. Therefore, we assessed 

the incidence and clinical characteristics of Fanconi syndrome 

during long-term ADV therapy of hepatitis B virus or human 

immunodeficiency virus infection. Methods We constructed a 

retrospective cohort composed of a total of 619 patients who 

took ADV for more than 30 days in a single tertiary teaching 

hospital between Jun 1994 and Dec 2013. Diagnosis of Fanconi 

syndrome required de novo appearance of at least two 

of three features: hypophosphatemia (below 2.5 mg/dL or 

decrement at least 0.5 mg/dL from baseline), hypouricemia 

(below 3.7 mg/dL or decline at least 0.5 mg/dL from baseline), 

or serum creatinine elevation (above 1.2 mg/dL or by at 

least 0.5 mg/dL from baseline). Results During the mean 27.6 

± 21.6 months ADV-treated period, 44 patients (7.1%) exhibited 

hypophosphatemia. Fourty-eight patients (7.8%) showed 

hypouricemia and 17 patients (2.7%) presented serum creatinine 

elevation. Among these patients, 6 patients (0.96%) 

developed Fanconi syndrome. Median time to development 

of Fanconi syndrome was 17.5 (range 15.17–35.07) months. 

All patients were treated with ADV for chronic hepatitis B. 

Conclusion Although Fanconi syndrome developed very rarely 

during low dose of ADV treatment, it occurred certainly in some 

patients. Physicians should carefully monitor the renal function 

and the level of serum phosphate and uric acid during the ADV 

treatment in particular after 1 year. 

Disclosures: 

The following authors have nothing to disclose: Sung Won Cho, Soo Yeon Cho, 

Choong Kyun Noh, Soon Sun Kim, Dukyong Yoon, Rae Woong Park, In-Whee 

Park 

2057 

The relationship between patient’s adherence to antiviral 

therapy and virological response in chronic hepatitis 

B and hepatitis C: Results of a 48-week observational 

study 

Iftihar Koksal, Seval Sonmez; Infectious Diseases, Black Sea Technical 

University Faculty of Medicine, Trabzon, Turkey 

Background:Patient’s adherence to antiviral drugs affects 

the success of chronic hepatitis B(CHB) and hepatitisC(CHC) 

treatment.In HBV, ‘non-adherence’ primarily refers to patientmissed 

doses.In HCV, in contrast,‘non-adherence’ primarily 

refers to dose reductions made by the clinician and early 

treatment discontinuation due to side-effects. Aim: To investigate 

the relationship between antiviral medication adherence 

and virological response in CHB & CHC patients. Materials– 

methods: Ninety-seven treated CHB (entecavir or tenofovir) 

and CHC (peg-IFN+ribavirin) patients were followed up for 

48 weeks in this prospective,non-interventional observational 

study.Patients were evaluated for virological response at the 

beginning of treatment and at the 4 th (CHC patients only), 

12 th , 24 th and 48 th weeks.Adherence was evaluated by counting 

prescribed drugs at each follow-up and through patient 

adherence questionnaires. Results: Twenty-one patients were 

excluded from the study.Of the 76 evaluated patients, 60.5% 

(n=46) had CHB and 39.5% (n=30) had CHC; 42.1% of the 

patients were women and 57.9% were men.Mean age was 

45.18±13.2.Based on data obtained from drug counts, 83.9% 

of the patients had at least one delay in drug administration at 

each visit.The most common causes of delayed drug administrations 

were omission (72.8%),running out of drugs and efforts 

to avoid side-effects.In addition, 92.6% of chronic hepatitis 

patients (n=68) exhibited adherence,and 92.1% of adherent 

and 62.5% of non-adherent patients responded to treatment. 

Adherent and non-adherent patient treatment response levels 

were statistically significant (p


equal to that of Pegasys 180μg. Results: 32 subjects entered 

the trial and 31 subjects completed study treatment. Groups 

were well generally matched (65% male, 12.5% HBeAg-negative, 

mean HBV DNA approximately 7.0 log10 IU/mL) at 

each group with HBV genotypes reflective of the population. 

rHSA/IFNα2a and Pegasys have shown antiviral activity and 

acceptable safety, tolerability. rHSA/IFNα2a was better tolerated 

than Pegasys such as neutrophil absolute value, white 

blood cell count, platelet count, hemoglobin and albumin 

decreased more at Pegasys group. Mean changes in serum 

HBV DNA were -1.32-2.13-1.10-2.48 log10 IU/mL for 600, 

750, and 900μg groups and Pegasys group after treatment. 

E antigen quantitation decreased more and normalization rate 

of ALT and AST were higher at rHSA/IFNα2a group. Half-life 

(t1/2) of rHSA/IFNα2a is 120 to 140 hours and duration of 

antiviral activity that indicate potential suitability for dosing 

intervals of 2 weeks or longer. Serum IFNα2a area under the 

curve and maximum concentration increased with dose escalating. 

IFNα2a Accumulation is not obvious at each group. 

Pharmacokinetics of the last dose except t1/2 and Kinetics of 

viral decline were similar between rHSA/IFNα2a 750μg and 

Pegasys. Conclusions: rHSA/IFNα2a was safe and well tolerated; 

declines in HBV DNA were similar to Pegasys evaluated. 

rHSA/IFNα2a 750μg has been selected for further hepatitis B 

clinical development. 

Disclosures: 

The following authors have nothing to disclose: Hong Zhang, Min Wu, Qingmei 

Li, xiaojiao li, Yanhua Ding, Junqi Niu 

2059 

Association between ALT flares and HBeAg loss and 

HBsAg decline in Patients with Chronic Hepatitis B 

during treatment with Tenofovir Disoproxil Fumarate or 

Adefovir Dipivoxil 

Patrick Marcellin 1 , Edward J. Gane 2 , Zahary Krastev 3 , Selim 

Gurel 4 , Geoffrey M. Dusheiko 5 , Anuj Gaggar 6 , Benedetta Massetto 

6 , Kyungpil Kim 6 , John F. Flaherty 6 , Mani Subramanian 6 , 

Harry L. Janssen 7 , Maria Buti 8 ; 1 Hôpital Beaujon, Clichy, France; 

2 Auckland City Hospital, Auckland, New Zealand; 3 University 

Hospital St. Ivan Rilsky, Sofia, Bulgaria; 4 Uludag Universitesi Tip 

Fakultesi, Bursa, Turkey; 5 Royal Free Hospital, London, United 

Kingdom; 6 Gilead Sciences, Inc., Foster City, CA; 7 University of 

Toronto, Toronto, ON, Canada; 8 Universitari Vall d’Hebron and 

Ciberehd, Barcelona, Spain 

Background and Aims: Increase in ALT levels (flares) are 

observed after the initiation of oral antiviral therapy and 

thought to be immune-mediated. We evaluated the association 

between ALT flares during the first 48 weeks of treatment with 

tenofovir disoproxil fumarate (TDF) or adefovir dipivoxil (ADV) 

and HBeAg loss and HBsAg decline. Methods: HBsAg levels 

were determined by Abbott Assay (linear range 0.05-250 

IU/mL; quantification test on undiluted samples, and at dilutions 

of 1:500 and 1:999) in 245 and 359 chronic infected 

HBeAg-positive and HBeAg-negative patients, respectively. ALT 

flare was defined as ALT >2x baseline and >5x upper limit 

of normal during the first 48 weeks of treatment. HBeAg loss 

and HBsAg decline were evaluated 24 weeks after a flare. 

Logistic regression analyses were performed to examine host 

and viral baseline factors (including viral load, HBV genotype, 

HBeAg status and HBsAg level) associated with flares. Significant 

factors in univariate analyses were examined in a multivariate 

model. Results: 23 HBeAg-positive patients (9%) and 5 

HBeAg-negative patients (1%) experienced ALT flares during 

the first 48 weeks of treatment. All ALT flares were considered 

not clinically significant except for one which was reported as 

a transient mild adverse event. Among HBeAg-positive patients, 

HBeAg loss and HBsAg decline were greater in patients who 

had ALT flares (Table, p


2060 

Antiviral treatment for chronic hepatitis B in Australia: a 

nationwide analysis of treatment uptake and prescribing 

trends according to individual antiviral agent 

Jennifer H. MacLachlan 1,2 , Nicole Allard 1,2 , Benjamin C. Cowie 1,2 ; 

1 WHO Collaborating Centre for Viral Hepatitis, Doherty Institute 

for Infection and Immunity, Melbourne, VIC, Australia; 2 Department 

of Medicine, Dentistry and Health Sciences, University of 

Melbourne, Melbourne, VIC, Australia 

Background: Antiviral treatment for chronic hepatitis B (CHB) 

is subsidized through Australia’s national health insurance 

scheme, Medicare, with all approved treatments funded for 

patients meeting specified virological and disease activity 

criteria. However, awareness and uptake of CHB therapy is 

low, and prescribing patterns and adherence to treatment 

guidelines have not been examined at a population level. 

Methods: Aggregate, population-level data regarding expenditure 

for CHB treatment through Medicare were obtained 

for all approved therapies (adefovir, entecavir, lamivudine, 

pegylated interferon, telbivudine, and tenofovir). Numbers of 

patients receiving therapy were derived from these data, and 

prescribing trends by individual agent were assessed for the 

period 2011-2013. Individualized data were also obtained 

for 2013 to examine prescribing patterns for combination 

antiviral therapy. Results: The number of patients treated for 

CHB in Australia increased from 9,000 in 2011 to 10,900 

in 2013, for an estimated proportion of all people living with 

CHB in 2013 of 5%. Patients receiving recommended first-line 

oral therapies represented an increasing majority, with those 

receiving entecavir increasing from 44.1% to 46.8% and tenofovir 

increasing from 26.3% to 34.6%. Usage of lamivudine 

and adefovir decreased, however together these drugs still 

represented 17.4% of prescribing expenditure in 2013. Very 

small numbers of patients received either telbivudine (36 months, consolidation 

duration>12 months and baseline HBV DNA level


mechanism triggered by introduction of small interfering RNA 

(siRNA). Tekmira’s clinically-validated lipid nanoparticle (LNP) 

platform enables effective delivery of siRNAs in vivo and in 

vitro. HDV-targeted siRNA combined with LNP technology may 

thus provide a novel opportunity to alleviate HDV pathogenesis. 

There is limited data regarding which of the HDV RNA 

species are susceptible to gene silencing. siRNAs targeting 

positive or negative strand HDV RNAs were designed and 

delivered to the Huh7-D12 stable HDV-expressing cell line 

using LNP technology. HDV RNAs were quantified using a 

branched DNA assay and nuclear/cytoplasmic fractionation, 

and HDAg protein was evaluated by ELISA. Rapid inhibitory 

effects were detected 1 day after LNP treatment, with HDAg 

mRNA-targeted siRNA showing a marked dose-dependent 

inhibitory effect on HDV positive strand RNAs but not on the 

negative strand viral genome. The nuclear/cytoplasmic fractionated 

RNA assay demonstrated that the cytosolic HDV RNAs 

were more susceptible to siRNA than nuclear HDV RNAs. In 

contrast to HDAg mRNA-targeted siRNAs that demonstrated 

gene silencing at day 1, genome-targeted siRNAs did not 

demonstrate gene silencing until day 4, hinting at differences 

in kinetics between the two siRNA targeting strategies. HDV-targeted 

siRNA-LNP effects were durable, with greater than 1-log 

reductions of viral RNAs maintained at even 21 days after a 

single treatment. Similarly, a 1-log reduction in HDAg protein 

was achieved out to day 12 so far and more extended time 

points are to be investigated. In conclusion, a direct HDV-targeted 

siRNA-LNP approach can effectively suppress positive 

and negative strand HDV RNAs and HDAg protein in vitro, and 

provides a promising novel strategy to treat HDV infection. The 

efficacy of direct HDV targeting relative to indirect effects from 

HBV gene silencing are currently under investigation. 

Disclosures: 

Xin Ye - Employment: Tekmira Pharmaceuticals Corporation 

Nicholas M. Snead - Employment: Tekmira Pharmaceuticals 

Trisha R. Barnard - Employment: Tekmira Pharmaceuticals Corporation 



The following authors have nothing to disclose: Ian MacLachlan 

2063 

Serum hepatitis B core-related antigen as a treatment 

predictor during pegylated interferon therapy in 

patients with HBeAg-positive chronic hepatitis B 

Natthaya Chuaypen 1 , Nawarat Posuwan 2 , Sunchai Payungporn 1 , 

Yasuhito Tanaka 3 , Yong Poovorawan 4 , Pisit Tangkijvanich 1 ; 1 Biochemistry, 

Chulalongkorn University, Bangkok, Thailand; 2 Chulalongkorn 

University, Bangkok, Thailand; 3 Virology and Liver unit, 

Nagoya City University Graduate School of Medical Sciences, 

Nagoya, Japan; 4 Paediatrics, Chulalongkorn University, Bangkok, 

Thailand 

Background: The role of quantitative serum hepatitis B core-related 

antigen (HBcrAg) in patients with chronic hepatitis B 

(CHB) receiving pegylated interferon (PEG-IFN) has never been 

investigated. The aims of this study were to assess the correlation 

of HBcrAg with intrahepatic covalently closed circular 

DNA (cccDNA), and compare its usefulness with quantitative 

serum HBsAg in patients with HBeAg-positive CHB during 

PEG-IFN therapy. Method: We retrospectively analyzed data 

of Thai patients with HBeAg-positive CHB treated with PEG- 

IFN for 48 weeks. Virological response (VR) was defined as 

HBeAg clearance and HBV DNA < 2,000 IU/mL at 24 weeks 

post treatment. Paired liver biopsies at weeks 0 and 48 were 

analyzed for cccDNA by real-time PCR. HBsAg and HBcrAg 

levels were assessed at weeks 0, 4, 12, 24, 48, and 72 by 

automated chemiluminescent immunoassays. HBV genotype 

was performed by direct sequencing. Results: A total of 46 

patients (31 male, mean age 33.2 years) were enrolled. 

The distribution of HBV genotypes B and C was 10.9% and 

89.1%, respectively. VR was achieved in 15 (32.6%) patients. 

Responders had higher cccDNA decline compared with non-responders 

(1.7±0.8 vs 0.4±1.1 log 10 

copies/cell, P=0.001), 

although baseline cccDNA were comparable between groups. 

Baseline HBsAg and HBcrAg were significantly correlated 

with cccDNA (Pearson correlation, r=0.450, P=0.013 and 

r=0.631, P20,000 IU/mL as the cut-off level, the 

negative predictive value (NPV) of achieving VR at weeks 12 

and 24 were 72.7% and 100%, respectively. For HBcrAg, the 

best cut-off level was log 10 

8.0 IU/mL, which provided NPV at 

week 12 and 24 of 89.5% and 100%, respectively. Conclusion: 

These results demonstrated that the convenient quantitative 

HBcrAg represented a good serum marker of intrahepatic 

cccDNA in patients with HBeAg-positive CHB, which was comparable 

with that of quantitative HBsAg. Monitoring HBcrAg 

levels during PEG-IFN therapy may be useful to identify patients 

with a very low probability of treatment response. 

Disclosures: 



Squibb 

The following authors have nothing to disclose: Natthaya Chuaypen, Nawarat 

Posuwan, Sunchai Payungporn, Yong Poovorawan, Pisit Tangkijvanich 

2064 

Encapsidation and secretion of HBV RNA can be inhibited 

by Core Inhibitors but not by Nucleoside Analogs 

Angela Lam, Suping Ren, Christine Espiritu, Mollie Kelly, Vincent 

Lau, Robert Vogel, George D. Hartman, Lalo Flores, Klaus Klumpp; 

Novira Therapeutics Inc., Doyleston, PA 

Background: Infectious HBV particles contain a partially double 

stranded DNA genome that is replicated in infected hepatocytes 

through reverse transcription of HBV pre-genomic RNA 

(pgRNA). HBV capsid, which encloses pgRNA and viral polymerase, 

is assembled from HBV core protein building blocks 

and is the site of HBV DNA synthesis. HBV core inhibitors 

are therefore able to potently inhibit HBV replication. In the 

current study, the effect of the HBV core inhibitor NVR-3891 on 

HBV RNA encapsidation and secretion was examined along 

with other core inhibitors and nucleoside analogs Lamivudine 

(LMV) or Tenofovir (TFV). Methods: Intracellular encapsidated 

HBV pgRNA was examined by Northern blot. HBV DNA and 

RNA levels were determined using Quantigene assays in HBV 

infected HepaRG cells and human serum samples. The nature 

and function of HBV RNA containing particles was evaluated 

using detergent and nuclease treatments, and analytical gradient 

centrifugation. Results: NVR-3891 induced HBV core 

protein mis-assembly in vitro and inhibited HBV replication 

with a mean EC 50 

of 1.9 mM in persistently infected differentiated 

HepaRG cells. Northern blot analysis of HBV infected 

cells showed that pgRNA encapsidation was inhibited by NVR- 

3891, while treatment of infected cells with nucleoside analogs 

led to an increase in encapsidated pgRNA. Analysis of 

the supernatants of infected cells and human sera from HBV 

infected patients showed high levels of HBV RNA containing


enveloped virus particles. Both HBV DNA and RNA containing 

particles fractionated similarly by density gradient centrifugation. 

The formation of these secreted RNA-containing viral particles 

could be blocked by treatment with core inhibitors, but 

not with nucleoside analogs. Conclusions: HBV RNA containing 

enveloped particles were observed in the supernatant of HBV 

infected cells and in the sera of HBV infected patients. Core 

inhibitors and nucleoside analogs could block HBV DNA production 

in HBV infected cells, but only core modulators could 

block the formation of HBV RNA containing particles. The more 

complete block of enveloped HBV particle formation (RNA and 

DNA particles) differentiates HBV core inhibitors from nucleoside 

analogs, which inhibit formation of only DNA-containing 

particles. Inhibitory effects on both HBV DNA and RNA particles 

could contribute to increased efficacy of HBV core inhibitor 

based treatment of chronic hepatitis B patients alone or in 

combination with nucleoside analogs. 

Disclosures: 

Angela Lam - Employment: Novira Therapeutics, Merck & Co 

Christine Espiritu - Employment: Novira Therapeutics 

Mollie Kelly - Independent Contractor: Novira Therapeutics 

George D. Hartman - Management Position: Novira Therapeutics 

Klaus Klumpp - Board Membership: Riboscience LLC; Employment: Novira Therapeutics 

Inc 

The following authors have nothing to disclose: Suping Ren, Vincent Lau, Robert 

Vogel, Lalo Flores 

2065 

Development and Recurrence of Hepatocellular Carcinoma 

in Patients with Chronic Hepatitis B Treated with a 

Nucleic Acid Analog 

Itaru Ozeki, Tomoaki Nakajima, Shuhei Hige, Yoshiyasu Karino, 

Joji Toyota; Department of Gastroenterology, Sapporo Kosei General 

Hospital, Sapporo, Japan 

Abstract Objectives: (1) To analyze the incidence of hepatocellular 

carcinoma (HCC) and factors contributing to carcinogenesis 

in patients treated with a nucleic acid analog (NA) 

and (2) to examine the background factors of HCC patients 

according to tumor recurrence. Materials and Methods: This 

study included 469 patients who received NA at our hospital 

for ≥1 year (305 who received entecavir for the first time 

and 164 who started lamivudine). Patients with a history of 

HCC or who received NA for


as proportions of resistances mutations maintain a relatively 

low level. Genotypic resistance to ETV was detected in all the 

partial responders with long-term therapy. The threshold related 

to substisutions requires corroboration by further studies with 

large samples to determine whether adapt the therapy.In addition 

to the common substitutions, the unknown amino acid substitutions, 

such as rtL145M/S, rtF151Y/L,rtR153Q, rtI224V, 

rtN248H, rtS223A, rtS256C,are also likely to influence the 

outcome of treatment. 

Disclosures: 

The following authors have nothing to disclose: Xiaxia Zhang, Minran Li, Ying 

Cao, Yu Zhang, Renwen Zhang, Yang Xu, Fang Li, Xiaoyuan Xu 

2067 

A Broad Spectrum Antibiotic Therapy As Empirical 

Treatment In Healthcare-Associated Infections Improves 

Survival In Cirrhotic Patients: A Randomized Trial 

Cristina Lucidi, Vincenza Di Gregorio, Barbara Lattanzi, Valerio 

Giannelli, Michela Giusto, Oliviero Riggio, Mario Venditti, Manuela 

Merli; Sapienza University of Rome, Rome, Italy 

Background and aims: An early diagnosis and an appropriate 

treatment of infections in cirrhosis are crucial due to their high 

morbidity and mortality. Multidrug-resistant (MDR) infections 

are growing in healthcare setting. Frequently, Healthcare-associated 

(HCA) infections are treated as community-acquired 

with a detrimental effect on survival. We aimed to prospectively 

evaluate for the first time in a randomized trial the effectiveness 

of a broad spectrum antibiotic treatment cirrhotic 

patients with HCA infections. Methods: Consecutive cirrhotic 

patients with HCA infections hospitalized in the last 3 years 

in our ward were enrolled. After culture sampling, patients 

were promptly randomized to empirically receive a standard 

or a broad spectrum antibiotic treatment (NCT01820026). The 

primary endpoint was the in-hospital mortality with the aim of 

66% reduction. Efficacy, sides effects, development of second 

infections and length of hospitalization was also considered. 

Treatment failure was followed by modification of therapies 

as appropriate. Results: As originally designed, 96 patients 

were randomized but 94 completed the treatment and were 

analyzed (two were excluded for incorrect randomization due 

to neoplastic ascites). Patients belonging to standard (48) and 

broad spectrum (46) groups were similar for demographic, 

clinical, and microbiological characteristics. The prevalence of 

MDR pathogens was similar in the 60 microbiologically documented 

infections (40% in standard vs 46% in broad spectrum 

group, p=ns). In-hospital mortality showed a 76% reduction 

in the broad spectrum vs standard group (respectively, 6% vs 

25%; p=0.01); the shift to another antibiotic treatment was not 

possible in about 20% of death patients. The standard therapy 

failed more often than the broad spectrum one (51 vs 18%, 

p=0.001). The length of the hospitalization was longer in the 

Standard Group (18 ± 15 days) than in the Broad Spectrum 

Group (12.3 ± 7 days) (p= 0.03). Only one patient showed 

a not severe side effect related to antibiotic treatment. Ten 

patients (12% of the group 1 and 9% of the group 2) developed 

a second episode of infection during the hospitalization 

with a similar prevalence of MDR of (60% in standard vs 

50% in broad spectrum, p=ns). Conclusions: A broad spectrum 

antibiotic therapy as empirical treatment in HCA infections 

improves survival in cirrhotic patients. This treatment resulted 

significantly effective, safe and cost-saving. 

Disclosures: 

The following authors have nothing to disclose: Cristina Lucidi, Vincenza Di 

Gregorio, Barbara Lattanzi, Valerio Giannelli, Michela Giusto, Oliviero Riggio, 

Mario Venditti, Manuela Merli 

2068 

Probiotic supplementation improves liver function but 

fails to restore neutrophil phagocytosis in stable cirrhosis. 

A randomized, double-blind, placebo-controlled 

study 

Angela Horvath, Bettina Leber, Bianca Schmerboeck, Monika 

Tawdrous, Astrid Hartl, Sandra Lemesch, Peter Fickert, Rudolf E. 

Stauber, Philipp Stiegler, Franziska Durchschein, Elisabeth Krones, 

Philipp Douschan, Gernot Zollner, Walter Spindelboeck, Karl 

Oettl, Doris Payerl, Vanessa Stadlbauer; Medical University Graz, 

Graz, Austria 

A multispecies probiotic was administered to cirrhotic patients 

in order to strengthen gut barrier function, decrease endotoxin 

load and ultimately restore phagocytic dysfunction of neutrophils 

that is commonly found in cirrhosis. Therefore, stable 

cirrhotic patients received either a mixture of Bifidobacteria bifidum 

and lactis, Lactobacili acidophilus, brevis, casei and salivarius, 

and Lactococcus lactis sp. (n=44) or a placebo (n=36) 

for 6 months. We found that during the intervention period 

the percentage of patients with Child-Pugh grade A increased 

significantly from 63% to 70% after three months and 66% 

after six months, only in the probiotic group. Of 16 patients 

in the probiotic group that started the study with Child-Pugh 

score 7 or higher, 6 improved their score after 6 months of 

probiotics, 7 did not change but only 3 deteriorated. Median 

Child-Pugh score decreased from 6 to 5 after three months of 

probiotics and MELD score decreased non-significantly from 

12 to 11 during the intervention period but returned to baseline 

6 months after intervention had ended. Gut permeability, 

measured with Lactulose-Mannitol ratio, improved significantly 

in both groups. With probiotics it remained significantly lower 

than baseline 6 months after the intervention has ended. On 

the other hand sucrose recovery, calprotectin and zonulin in 

stool and serum diamine oxidase stayed unchanged in both 

groups. Endotoxin levels decreased non-significantly with probiotics 

but similar changes occurred in the control group. Cytokine 

levels as well as soluble CD14 and LPS binding protein 

stayed unchanged in both groups. Probiotic supplementation 

did not have a significant influence on neutrophil phagocytosis. 

At baseline both groups had an intact phagocytic capacity but 

a significantly increased amount of non-phagocytic neutrophils 

compared to controls. Over the course of the study the amount 

of non-phagocytic cells remained constant while phagocytic 

capacity decreased to 60% of the baseline value. During the 

first 6 months of the study 1 severe infection occurred in the 

placebo group, none in the probiotic group. After intervention 

had ended 5 severe infections were reported in the probiotic 

group and 4 in the placebo group. In conclusion, probiotic 

supplementation is a save method to improve liver function in 

stable cirrhosis. However, its influence on gut barrier function 

and bacterial translocation in cirrhotic patients seem to be minimal. 

Independent of the intervention, patients gradually developed 

significant impairment in neutrophil phagocytosis over 

the course of one year and with this loss of function a higher 

incidence of severe infections occurred. 

Disclosures: 

Angela Horvath - Grant/Research Support: Instutut Allergosan 

Peter Fickert - Consulting: Falk Foundation, Falk Foundation, Falk Foundation, 

Falk Foundation; Speaking and Teaching: Roche Austria, Gilead Austria, MSD 

Austria, MERCK Austria, Roche Austria, Gilead Austria, MSD Austria, MERCK 

Austria, Roche Austria, Gilead Austria, MSD Austria, MERCK Austria, Roche 

Austria, Gilead Austria, MSD Austria, MERCK Austria 

Rudolf E. Stauber - Advisory Committees or Review Panels: Gilead, Janssen-Cilag, 

AbbVie, BMS; Grant/Research Support: MSD; Speaking and Teaching: Merz 

The following authors have nothing to disclose: Bettina Leber, Bianca Schmerboeck, 

Monika Tawdrous, Astrid Hartl, Sandra Lemesch, Philipp Stiegler, Franziska 

Durchschein, Elisabeth Krones, Philipp Douschan, Gernot Zollner, Walter 

Spindelboeck, Karl Oettl, Doris Payerl, Vanessa Stadlbauer


2069 

Impact of genetic variation of the AVP1a receptor on 

the presence of circulatory failure in patients with acute 

decompensation of liver cirrhosis or acute-on-chronic 

liver failure 

Annarein J. Kerbert 1 , Jelte Schaapman 1 , Johan van der Reijden 1 , 

Amorós Àlex 2 , Aiden McCormick 3 , Bart van Hoek 1 , Vicente 

Arroyo 4 , Pere Gines 4 , Rajiv Jalan 5 , Victor Vargas 6 , Rudolf E. 

Stauber 7 , Hein W. Verspaget 1 , Minneke Coenraad 1 ; 1 Gastroenterology-Hepatology, 

Leiden University Medical Center, Leiden, 

Netherlands; 2 Data Management Center, CLIF consortium, Barcelona, 

Spain; 3 Gastroenterology-Hepatology, St Vincent’s University 

Hospital, Dublin, Ireland; 4 Liver Unit, Hospital Clinic, University 

of Barcelona, Barcelona, Spain; 5 Gastroenterology-Hepatology, 

University College London, London, United Kingdom; 6 Liver Unit, 

Hospital Vall d’Hebron, CIBERehd, Barcelona, Spain; 7 Internal 

Medicine, Medical University of Graz, Graz, Austria 

Background & aims: Acute-on-chronic liver failure (ACLF) 

is defined as an acute decompensation of former stable 

chronic liver disease (AD) accompanied by the presence of 

organ failure and a high risk of short-term mortality. Systemic 

hemodynamic derangement and activation of endogenous 

vasoconstrictor systems are thought to contribute to the pathogenesis. 

Arginine vasopressin is a key-regulator in hemodynamic 

homeostasis and mediates splanchnic vasoconstriction 

through the arginine vasopressin 1a receptor (AVP1aR). Aim 

of the present study was to assess whether genetic variation of 

AVP1aR is associated with the presence of circulatory failure 

in patients with AD or ACLF. Methods: Eight single nucleotide 

polymorphisms (SNPs) of AVP1aR with possible clinical relevance 

were identified. From 824 cirrhotic patients admitted for 

AD, clinical, laboratory and survival data were retrieved from 

the CANONIC database. Presence of circulatory failure was 

defined as a mean arterial blood pressure (MAP) < 70 mmHg 

or the use of vasopressors. ACLF was defined according to 

the CLIF Consortium Organ Failure score. All patients were 

genotyped for all eight SNPs using polymerase chain reaction 

(PCR) followed by restriction fragment length polymorphism 

or PCR allele-specific amplification primers. Fisher’s exact test 

and linear regression analysis were used to test for association 

between allele frequencies and dichotomous and continuous 

variables respectively. Results are shown as mean ± SD. P< 

0.05 was considered statistically significant. Results: Patients 

with ACLF (n=184) had a significantly lower MAP as compared 

to patients without ACLF (80 ± 13 vs. 84 ± 12 mmHg, 

p< 0.001). The use of vasopressors was also significantly more 

frequent in patients with ACLF as compared to those without 

ACLF (19.0% vs. 2.9%, p< 0.001). Circulatory failure was 

present in 61 out of 824 patients, of whom 44 fulfilled the 

criteria of ACLF. A C>T mutation in SNP rs7308855 showed 

a significant association with the presence of circulatory failure 

in patients with AD (p= 0.025) and a clear trend towards the 

presence of circulatory failure in patients with ACLF (p= 0.085). 

A trend was also found for a T>A mutation in SNP rs7298346 

to be associated with the presence of circulatory failure in 

patients with AD (p= 0.062). In addition, this mutation showed 

a significant association with the presence of circulatory failure 

in the subgroup of patients with ACLF (p= 0.046). Conclusions: 

Single nucleotide polymorphisms in the AVP1a receptor are 

associated with the presence of circulatory failure in patients 

with acute decompensation of liver disease and ACLF. 

Disclosures: 



Vicente Arroyo - Speaking and Teaching: GRIFOLS 

Pere Gines - Advisory Committees or Review Panels: Ferring, Ikaria; Grant/ 

Research Support: Sequana Medical, Grifols 



Rudolf E. Stauber - Advisory Committees or Review Panels: Gilead, BMS; Grant/ 

Research Support: Abbvie, MSD; Speaking and Teaching: Merz 

The following authors have nothing to disclose: Annarein J. Kerbert, Jelte Schaapman, 

Johan van der Reijden, Amorós Àlex, Aiden McCormick, Victor Vargas, 

Hein W. Verspaget, Minneke Coenraad 

2070 

Prevalence, risk factors and outcome of infections by 

multidrug resistant bacteria (MDR) in a liver intensive 

care unit (ICU): a prospective study 

Amrish Sahney 1 , Cyriac A. Philips 1 , Rakhi Maiwall 1 , Ashok 

Choudhary 1 , Lalita G. Mitra 2 , Ankur Jindal 1 , Manoj Kumar 1 , Kapil 

D. Jamwal 1 , Vikram Bhatia 1 , Vikas Khillan 3 , Shiv K. Sarin 1 ; 1 Hepatology, 

ILBS, New Delhi, India; 2 Critical care, ILBS, New Delhi, 

India; 3 Microbiology, ILBS, New Delhi, India 

Background & Aim:There is limited data on prevalence, predictors 

and impact on extra hepatic organ failure and mortality 

due to MDR bacterial infections in critically ill cirrhotic 

patients. Methodology: We prospectively studied 522 cirrhoticpatients 

admittedto dedicated liver ICU during March 

to October 2014. Organ failure was defined as: Kidney failure: 

SCr ≥2mg% or need for dialysis, cerebral failure: Grade 

3 or 4 hepatic encephalopathy, Circulation failure: shock, & 

Lung failure: PaO2/FiO2


Disclosures: 

The following authors have nothing to disclose: Amrish Sahney, Cyriac A. Philips, 

Rakhi Maiwall, Ashok Choudhary, Lalita G. Mitra, Ankur Jindal, Manoj Kumar, 

Kapil D. Jamwal, Vikram Bhatia, Vikas Khillan, Shiv K. Sarin 

2071 

The macrophage activation markers sCD206 and 

sCD163 predict mortality in patients with liver cirrhosis 

without or with acute-on-chronic liver failure (ACLF) 

Henning Gronbaek 1 , Sidsel Rødgaard-Hansen 2 , Niels Kristian 

Aagaard 1 , Elisabet García 3 , Hendrik V. Vilstrup 1 , Søren K. 

Moestrup 4 , Vicente Arroyo 3 , Holger J. Møller 2 ; 1 Department of 

Hepatology & Gastroenterology, Aarhus University Hospital, Aarhus, 

Denmark; 2 Department of Clinical Biochemistry, Aarhus University 

Hospital, Aarhus, Denmark; 3 Liver Unit, Hospital Clinic, 

University of Barcelona, Barcelona, Spain; 4 Department of Biomedicine, 

Aarhus University, Aarhus, Denmark 

Introduction: Activation of liver macrophages plays a key role 

in liver and systemic inflammation and may be involved in 

the development and prognosis of ACLF. We therefore measured 

the macrophage activation markers soluble sCD206 and 

sCD163 in plasma and related them to the short- (1-3 months) 

and long-term (6 months) mortality in the cirrhosis patients of the 

CANONIC study. Methods: 86 patients had no ascites and no 

ACLF, 580 had ascites but no ACLF, and 100, 66 and 19 had 

ACLF-grade I (ACLF-1), ACLF-II, and ACLF-III, respectively. The 

patients’ clinical course was registered and their MELD, CLIF-C 

Acute Decompensation (AD), and ACLF (s)scores computed at 

study entry. Results: We found a stepwise increase (p


2073 

The Epidemiology and Clinical Associations of Budd- 

Chiari Syndrome. A Review of 507 Admissions from the 

2012 National Inpatient Sample 

Jennifer Saad 1 , Shawn Shah 1 , Joseph Shatzel 1 , Harley Friedman 1 , 

Neil Volk 1 , Rolland C. Dickson 2 ; 1 Internal Medicine, Dartmouth 

Hitchcock Medical Center, Lebanon, NH; 2 Gastroenterology, Dartmouth 

Hitchcock Medical Center, Lebanon, NH 

Background: Budd-Chiari syndrome (BCS) is a rare condition 

resulting from obstruction of the hepatic venous outflow tract 

and is strongly associated with hypercoagulable states. Large 

series on BCS are scarce, so epidemiological data has been 

derived primarily from reported case series. The aim of this 

study was to evaluate the epidemiology, common clinical associations, 

and mortality of patients admitted with BCS from a 

large national database. Methods: Using the 2012 National 

Inpatient Sample (NIS), admissions with an International Classification 

of Diseases, 9 th Revision, Clinical Modification code 

for BCS were extracted, and correlated with age, gender, 

length of stay, mortality and commonly associated diagnoses. 

Results: Of the 7,296,968 unweighted admissions in the 2012 

NIS, 507 were associated with BCS (prevalence 0.007%). 

BCS was more common in women (55.8%) with a mean age 

of 47 years (range 0-90). The average length of stay was 10 

days (range 0-125) with an in-hospital mortality of 6.5%. Of 

BCS admissions, 23.08% had cirrhosis of the liver, 16.77% 

had portal HTN, 8.68% had both cirrhosis of the liver and 

portal HTN, 16.3% had a hepatocellular malignancy (8.3% 

primary, 8.1% metastatic) and 8.9% had an acquired or inherited 

thrombophilia. Overall, 37.5% of patients with BCS had 

a new or previously diagnosed malignancy. Myeloproliferative 

or bone marrow disorders were associated with 8.1% of 

BCS admissions including polycythemia vera (3.8%), essential 

thrombocytosis (1.6%), paroxysmal nocturnal hemoglobinuria 

(1.4%), myelofibrosis (0.8%) and chronic myelogenous leukemia 

(0.6%). 28% of admissions had an acute thrombus at an 

additional anatomic site (9.9% superficial or deep veins, 8.9% 

inferior vena cava, 6.1% pulmonary vein and 3.2% portal 

vein). There was no significant difference in the age, mortality 

or length of stay between males and females with BCS. Males 

were more likely to have a primary hepatocellular malignancy 

(OR 3.97, 95% CI 1.95, 8.10, p


2075 

Circulating interleukin 6, 10 and 17 as prognostic 

markers in patients with liver cirrhosis 

Josiane Fischer 1 , Telma E. Silva 1 , Pedro E. Soares e Silva 1 , Bruno 

S. Colombo 1 , Letícia M. Wildner 2 , Maria Luiza Bazzo 2 , Tania 

S. Frode 2 , Silvana Vigil de Melo 2 , Julia S. Rosa 2 , Esther B. Dantas-Correa 

1 , Janaína L. Narciso-Schiavon 1 , Leonardo L. Schiavon 1 ; 

1 Internal Medicine, Division of Gastroenterology, Federal University 

of Santa Catarina, Florianópolis, Brazil; 2 Department of Clinical 

Analysis, Federal University of Santa Catarina, Florianopolis, 

Brazil 

Introduction: Activation of inflammatory system is present from 

the early stages of cirrhosis and it is associated with elevated 

levels of cytokines. However, data about the prognostic significance 

of circulating cytokines in liver cirrhosis is still lacking. 

We sought to investigate the prognostic significance of IL-6, 

IL-10 and IL-17 in patients with stable cirrhosis and in subjects 

admitted for acute decompensation (AD) of cirrhosis. Methods: 

This prospective study included two cohorts: (1) stable cirrhosis 

attended in the Outpatient Clinic (n = 118), and (2) subjects 

hospitalized for AD (n = 130). Thirty healthy subjects served as 

control group. The acute-on-chronic liver failure (ACLF) criteria 

were applied according to the EASL-CLIF Consortium. Results: 

IL-6 and IL-10 levels were higher in both groups of patients with 

cirrhosis as compared to control group and also in patients 

with AD in relation to stable cirrhosis (P < 0.05). In stable 

cirrhosis, during a median follow-up of 17 months, an event 

(hospitalization, death or liver transplantation) occurred in 26 

patients and was associated with higher IL-6 (3.56 pg/mL vs. 

2.13 pg/mL, P = 0.013) and IL-10 (0.54 pg/mL vs. 0.22 pg/ 

mL, P = 0.021), but not IL-17 levels. In the hospitalized cohort, 

39 patients died after 90 days of follow-up. Logistic regression 

analysis showed that death in AD cohort was independently 

associated with ascites (OR 6.286, 95% CI 1.826 – 21.635; 

P = 0.004), MELD (OR 1.300, 95% CI 1.175 – 1.439; P 

< 0.001) and IL-6 (OR 1.002, 95% CI 1.000 – 1.004, P = 

0.029). The AUROC of IL-6 to predict 90-day mortality was 

0.779 ± 0.046 and the Kaplan–Meier survival probability 

was 90.0% for IL-6 < 21 pg/mL and 46.7% for IL-6 ≥ 21 

pg/mL (P < 0.001). Cytokine levels were evaluated for the 

prediction of bacterial infection. Regression analysis showed 

that bacterial infection diagnosed during the first 48 hours 

after admission was associated with IL-6, CRP and ascites. IL-6 

exhibited higher AUROC than CRP for predicting bacterial 

infection (0.831 ± 0.043 vs. 0.763 ± 0.048, respectively). 

Higher IL-6 levels were observed in ACLF patients even in the 

absence of bacterial infection whereas IL-10 was higher only 

in subjects with infection-related ACLF. Cytokines levels were 

reassessed at the third day of hospitalization in 74 subjects. 

No differences between admission and third-day levels were 

noted for IL-6. Lower IL-10 levels were observed at third day 

regardless of the presence of ACLF or death during follow-up. 

However, IL-17 levels dropped significantly only in those who 

died during follow-up. Conclusion: Circulating IL-6, IL-10 and 

IL-17 are of prognostic value in patients with cirrhosis. 

Disclosures: 

The following authors have nothing to disclose: Josiane Fischer, Telma E. Silva, 

Pedro E. Soares e Silva, Bruno S. Colombo, Letícia M. Wildner, Maria Luiza 

Bazzo, Tania S. Frode, Silvana Vigil de Melo, Julia S. Rosa, Esther B. Dantas-Correa, 

Janaína L. Narciso-Schiavon, Leonardo L. Schiavon 

2076 

Systemic inflammatory response syndrome (SIRS) in 

cirrhotic patients with and without bacterial infections: 

clinical characteristics and correlation with liver and 

kidney function and prognosis 

Caroline B. Souza, Livia B. Victor, Camila M. Alcântara, Tatiana 

Valdeolivas, Vanessa L. Zenatti, Daniela M. Mariz, Zulane D. 

Veiga, Emilia Ahmed, Flavia F. Fernandes, Gustavo Pereira; Hospital 

Geral de Bonsucesso, Rio de Janeiro, Brazil 

SIRS has been associated with morbidity and mortality in cirrhosis. 

Although commonly described in infections, it’s also 

observed in patients without it. Differences in clinical data, incidence 

of complications and survival between these two groups 

have not been studied so far. Objective: Analyze differences 

in clinical characteristics, diagnostic criteria, liver and kidney 

function, complications and survival in cirrhotics with SIRS 

associated or not with infections. Methods: 256 patients were 

evaluated. Diagnosis of SIRS was established within 48 hours 

of admission. Patients were classified as having sepsis (SIRS 

associated with infections) or SIRS without infection. Additionally 

we compared these groups with patients with bacterial 

infections without SIRS and a control group, composed of cirrhotic 

patients without neither SIRS nor infections. Results: Prevalence 

of SIRS was 35%. SIRS was diagnosed at admission 

in 51 and developed within 48h in 39 patients. The majority 

of patients fulfilled 2 diagnostic criteria (83%) of which leucocytes 

and temperature were the most common (69 and 62%). 

Sepsis was diagnosed in 55 patients and SIRS without infections 

in the remaining 35. Patients with sepsis more frequently 

had 3 or more diagnostic criteria for SIRS (22 vs. 9%, p=0.1) 

and higher leucocyte count (9.6 ±8.1 vs 6.2±4.6, p=0.006). 

Patients with sepsis had worst liver and kidney function, as 

evidenced by lower albumin and higher INR and creatinine 

values. Complications of cirrhosis were equally distributed in 

both groups, except for higher frequency of gastrointestinal 

bleeding in SIRS group (29 vs 6%, p=0.003). Acute-on-Chronic 

Liver Failure (ACLF) was more frequent in sepsis group (46 vs 

13%, p=0.001). The probability of survival at 28 and 90 days 

was 79 and 68%. Patients with Sepsis had lower survival than 

patients with SIRS both at 28 days (62 vs. 86%) and 90 days 

(53% vs. 74%), p


2077 

The NDP52 Val248Ala variant increases the risk for 

spontaneous bacterial peritonitis in patients with alcoholic 

liver cirrhosis. 

Philipp Lutz 1,2 , Benjamin Krämer 1,2 , Dominik J. Kaczmarek 1,2 , 

Marc P. Hübner 3,2 , Bettina Langhans 1,2 , Beate Appenrodt 4 , Frank 

Lammert 4 , Jacob Nattermann 1,2 , Achim Hoerauf 3,2 , Christian 

P. Strassburg 1,2 , Ulrich Spengler 1,2 , Hans Dieter Nischalke 1,2 ; 

1 Department of Internal Medicine I, Bonn, Germany; 2 German 

Center for Infection Research, Bonn, Germany; 3 Institute for Medical 

Microbiology, Immunology and Parasitology, University of 

Bonn, Bonn, Germany; 4 Department of Medicine II, Saarland University 

Medical Center, Homburg, Germany 

Aim: Spontaneous bacterial peritonitis (SBP) is frequently a 

fatal infection in patients with advanced liver cirrhosis. We 

investigated if NDP52 (nuclear dot protein 52 kDa), a negative 

regulator of toll-like receptor signalling and an autophagy 

adaptor protein, might be involved in the development of SBP. 

Methods: Two cohorts comprising 152 (derivation cohort) and 

198 patients (validation cohort) with liver cirrhosis and ascites 

as well as 168 healthy controls were genotyped for the NDP52 

Val248Ala variant and stratified for the occurrence of SBP. 

Results: Overall, 57 (38%) patients in the derivation cohort 

and 77 (39%) in the validation cohort had SBP. Cirrhosis was 

due to alcohol abuse in 57% of the derivation and 66% of the 

validation cohort. NDP52 genotype distribution was consistent 

with Hardy-Weinberg equilibrium. In patients with alcoholic 

cirrhosis, patients with SBP had an increased frequency of 

the NDP52 minor variant in the derivation (p=0.04) and in 

the validation cohort (p=0.01). In patients with liver cirrhosis 

of non-alcoholic etiology, the frequency of the NPD52 minor 

variant was comparable between patients with and without 

SBP (11.3% versus 12.2%). Multivariate analysis confirmed 

the NDP52 minor variant (odds ratio 4.7, p=0.002) and the 

TLR2 -16934 TT variant (odds ratio 2.5, p=0.008) as independent 

risk factors for SBP in alcoholic liver disease. Analysis 

of the interaction between the TLR2 and the NDP52 risk 

variant revealed that patients carrying both risk variants had 

a particularly high risk to acquire SBP. Conclusion: Presence 

of the NPD52 minor variant is a risk factor for SBP in patients 

with alcoholic cirrhosis, indicating that autophagy might be 

involved in the pathogenesis of SBP. 

Disclosures: 



The following authors have nothing to disclose: Philipp Lutz, Benjamin Krämer, 

Dominik J. Kaczmarek, Marc P. Hübner, Bettina Langhans, Beate Appenrodt, 

Frank Lammert, Jacob Nattermann, Achim Hoerauf, Ulrich Spengler, Hans Dieter 

Nischalke 

2078 

Assessment and relevance of coagulation disorders in 

patients with acute-on-chronic liver failure during Systemic 

Inflammatory Response (SIRS) and development of 

sepsis 

Madhumita Premkumar, Priyanka Saxena, Roshni Mirza, Sukriti 

Sukriti, Chhagan Bihari, Ashok Choudhary, Chandan K. Kedarisetty, 

Shiv K. Sarin; Hepatology, Institute of Liver and Biliary Sciences, 

Delhi, India 

Background: Coagulation system is rebalanced in cirrhosis.The 

status of baseline coagulation disturbances in ACLF and their 

alterations with development of sepsis are largely unknown. 

Aim: To study the dynamic changes in coagulation profile in 

ACLF, and their correlation with evolving SIRS and sepsis. 

Patients and Methods : Of the 243 consecutive patients with 

ACLF (APASL criteria), 114 with no evidence of sepsis were 

recruited (mean age 44.3±11.7 yr, males 90%). Predominant 

etiology for underlying liver disease was ethanol (63.1%). 

Patients were evaluated by tests like INR and thromboelastography 

(TEG), Sonoclot, and coagulation factor assays. 

These results were compared with 25 controls. At presentation 

40/114 (35.1%) had SIRS. Twenty eight (24.5%) developed 

sepsis by day 3 and 52/112 (56.1%) by day 7. Only 11 /74 

(14.8%) developed sepsis de novo without prior SIRS. SIRS at 

presentation posed a mortality risk of 48%. A deranged TEG 

at presentation was a predictor of bleeding (OR 2.1, p=0.05) 

and mortality (OR 3.1, p=0.05).Platelet count and functions (as 

predicted by sonoclot) were significantly lower in ACLF patients 

with sepsis at day 3 and 7. INR at baseline was associated 

with a deranged R and K component of TEG (p=0.05) and 

ACT of Sonoclot (p=0.02). Baseline ACT, CR and PF (sonoclot 

parameters), were associated with development of SIRS at day 

3 (p=0.013) and sepsis at day 7( p=0.012). Likewise R and 

K of TEG (p=0.064) predicted sepsis. Coagulation assays for 

protein C,and antithrombin III were lower in all ACLF patients, 

when compared with controls, but did not change with sepsis. 

Factor 8 levels were significantly increased in all ACLF 

subgroups. Conclusions: Coagulation abnormalities in ACLF 

are associated and correlate with an increased tendency to 

bleed, risk of sepsis and mortality. Coagulation parameters 

such as INR, TEG and Sonoclot are affected by presence of 

SIRS. Sonoclot parameters at baseline can serve as predictors 

of development of sepsis and poor outcome 

Table 1: Coagulation parameters of ACLF when compared with 

those with sepsis at day 3 and day 7. 

ACT :Activated coagulation time 

Disclosures: 

The following authors have nothing to disclose: Madhumita Premkumar, Priyanka 

Saxena, Roshni Mirza, Sukriti Sukriti, Chhagan Bihari, Ashok Choudhary, Chandan 

K. Kedarisetty, Shiv K. Sarin 

2079 

Characteristics and outcome of Severe Alcoholic Hepatitis-related 

Acute-on-Chronic Liver Failure Syndrome 

Alexia Cornillie, Eric Trépo, Delphine Degré, Jonas Schreiber, 

Antonia Lepida, Christophe Moreno, Thierry Gustot; Gastroenterology 

and Hepato-Pancreatology, Erasme Hospital, Université 

Libre de Bruxelles, Brussels, Belgium 

Background & Aims: Although active alcoholism is known to 

be a potential trigger for Acute-on-Chronic Liver Failure (ACLF), 

place of severe alcoholic hepatitis (AH) as precipitating event is 

currently unknown. We assessed characteristics, outcome and 

predictive factors for the development of ACLF in patients with 

severe biopsy-proven alcoholic hepatitis (AH). Methods: We 

prospectively followed 145 consecutive biopsy-proven severe 

AH (mDF≥32) episodes for 6 months. We retrospectively 

reviewed ACLF diagnosis, grades (based on CANONIC criteria) 

and prognostic scores, CLIF-C Organ Failure score (OFs) 

and CLIF-C ACLFs (Jalan et al. J Hepatol 2014;61:1038-47) at 

the time of liver biopsy (baseline) and at different time points


during 3-month follow-up. Results: At baseline, ACLF was diagnosed 

among 52% of severe AH episodes (17% of ACLF grade 

1 (ACLF-1), 16% of ACLF-2 and 19% of ACLF-3). Compared 

to AH episodes without ACLF at baseline, AH episodes with 

ACLF had a higher 28-day and 90-day transplant-free mortality 

rate (54 vs. 10% and 68% vs. 18% respectively, p


ease diagnosis (p=0.00001, Odds Ratio 0.22 and p=0.008, 

OR 2.0 respectively) Conclusion: We found that sarcopenia 

has a higher prevalence among cirrhotic patients undergoing 

liver transplantation at our Institution than reported in the 

literature. Interestingly, obesity was found to be a significant 

predictor of pre-transplant sarcopenia in this cohort 

Disclosures: 

The following authors have nothing to disclose: Trushar Patel, Valery Vilchez, 

Rashmi T. Nair, Jennifer Watkins, Anna Christina Dela Cruz, Roberto Gedaly, 

Terrence Barrett 

2082 

Liver Cirrhosis Is Strongly And Independently Associated 

With Mortality In Patients With Bacterial Endocarditis: 

Results Of A Case Control Multicenter Study Of 202 

Cases 

Jean françois D. Cadranel 1 , Isabelle Ollivier-Hourmand 2 , Salah 

Zerkly 3 , Christophe Bureau 4 , Thierry Thevenot 5 , Patrice Cacoub 6 , 

Armand Garioud 1 , Gilles Macaigne 7 , Laurent Alric 8 , Vincent Jouannaud 

4 , Hortensia Lison 1 , Carine Chagneau-Derrode 9 , Alexandre 

Pariente 10 , Agnes Pelaquier 11 , Marc Bourlière 13 , Xavier Causse 12 , 

Manon Allaire 2 , Jean-Baptiste Nousbaum 14 , Jerôme Dumortier 15 , 

Alexandre Louvet 16 , Isabelle Rosa 17 , Nathalie Ganne-Carrié 18 , 

Jerôme Gournay 19 , Hélène Blasco-Perrin 4 , Teresa Antonini 20 , Laurent 

Spahr 21 , Jean-Pierre Bronowicki 22 , Christine Silvain 9 , Xavier 

Amiot 23 , Vincent Di Martino 5 , Bruno Lesgourgues 24 , Jean-Didier 

Grangé 23 , Jean-Marie Péron 4 , Didier Samuel 25 , Xavier Adhoute 13 , 

Hervé Hagège 17 , Philippe Mathurin 16 , Jacques Denis 26 , Pierre 

Iaria 27 , Thong Dao 2 ; 1 Hepatogastoenterology department, 

GHPSO Creil, Creil, France; 2 Hepatogastroenterology Unit, CHU, 

Caen, France; 3 Epidemiology Unit, CHU, Amiens, France; 4 Hepatogastroenterology 

Unit, CHU, Toulouse, France; 5 Hepatogastroenterology 

Unit, CHU, Besançon, France; 6 Internal Medicine Unit, 

CHU Pitié-Salpêtrière, Paris, France; 7 Hepatogastroenterology 

Unit, CH, Lagny-sur-Marne, France; 8 Internal medicine Unit, CHU, 

Toulouse, France; 9 Hepatogastroenterology Unit, CHU, Poitiers, 

France; 10 Hepatogastroenterology Unit, CH, Pau, France; 11 Hepatogastroenterology 

Unit, CH, Montélimar, France; 12 Hepatogastroenterology 

Unit, CHR, Orléans, France; 13 Hepatogastroenterology 

Unit, CH Saint-Joseph, Marseille, France; 14 Hepatogastroenterology 

Unit, CHU, Brest, France; 15 Hepatogastroenterology Unit, 

CHU, Lyon, France; 16 Hepatogastroenterology Unit, CHU, Lille, 

France; 17 Hepatogastroenterology Unit, CHIC, Créteil, France; 

18 Hepatogastroenterology Unit, CHU Jean Verdier, Bondy, France; 

19 Hepatogastroenterology Unit, CHU, Nantes, France; 20 Centre 

Hépato-Biliaire, Villejuif, France; 21 Hepatogastroenterology Unit, 

Geneva Hospitals, Geneva, Switzerland; 22 Hepatogastroenterology 

Unit, CHU, Nancy, France; 23 Hepatogastroenterology Unit, 

CHU Tenon, Paris, France; 24 Hepatogastroenterology Unit, CH, 

Montfermeil, France; 25 Hepatogastroenterology Unit, Centre Hépato-Biliaire, 

Villeuif, France; 26 Hepatogastroenterology Unit, CH Sud 

Francilien, Corbeil-Essonnes, France; 27 Cardiology Unit, GHPSO, 

Creil, France 

Background and Aims: Bacterial endocarditis (BE) is severe in 

patients (pts) with liver cirrhosis (LC); data rely on uncontrolled 

studies. Aims of this study were to compare clinical presentation 

and mortality of BE in LC pts with those of controls (CT) without 

cirrhosis matched for sex, age and diabetes. Methods: Medical 

charts for all LC pts with BE seen in 23 liver units (2000-2013) 

were reviewed and compared with those of CT. Short-term mortality 

was analyzed by univariate and logistic regression analysis. 

Results: 101 BE in LC pts and 101 BE in CT were analyzed. 

LC pts: 63.2 [42-87] years, CT: 65 [46-93]; 145 M (72.1%). 

LC causes: alcohol: 78 (67.2%), viral: 17 (14.6%), NAFLD 14 

(12%). Child-Pugh scores: A: 8pts (8.8%), B: 39 pts(42.9%), C: 

44 pts(48.4%). In LC pts: total bilirubin was 67.8μmol/l±77.7, 

prothrombin time (PT) 52.7%±18.1, serum albumin 25.2g/ 

l±5.9, serum creatinine (SC) 123.5μmol/l±103.6. Blood cultures 

were positive in 181 pts (91.4%). 39 LC pts (40.2%) 

and 52 CT (53.6%) had a pre-existing heart disease (NS). 

At diagnosis, 43 LC (47.7%) and 31 CT (34.0%) exhibited 

heart failure (NS). Severe sepsis or septic shock was noted 

in 33 LC pts (33.3%) and 23 CT (23.0%) (NS). The source 

of contamination was digestive, urinary, or cutaneous (DUC) 

in 41 LC pts and in 21 CT (OR 2.4 [1.1–5.1], p


The following authors have nothing to disclose: Jean françois D. Cadranel, Salah 

Zerkly, Thierry Thevenot, Armand Garioud, Gilles Macaigne, Vincent Jouannaud, 

Hortensia Lison, Carine Chagneau-Derrode, Agnes Pelaquier, Manon Allaire, 

Jean-Baptiste Nousbaum, Jerôme Dumortier, Alexandre Louvet, Isabelle Rosa, 

Jerôme Gournay, Hélène Blasco-Perrin, Teresa Antonini, Laurent Spahr, Christine 

Silvain, Bruno Lesgourgues, Jean-Didier Grangé, Jean-Marie Péron, Xavier 

Adhoute, Hervé Hagège, Pierre Iaria 

2083 

Hispanic ethnicity is associated with severe sepsis 

among infected patients with end-stage liver disease 

Vinay Sundaram 1 , Jason J. Xu 1 , Christie Jeon 1 , David S. Goldberg 

2 ; 1 Cedars-Sinai Medical Center, Los Angeles, CA; 2 Medicine, 


Background & Aims: The risk and prognosis for severe sepsis 

varies by race and ethnicity in the general population. We 

aimed to determine whether these disparities apply in patients 

with end-stage liver disease. Methods: We studied the Nationwide 

Inpatient Sample, 2009-2011. The cohort included 

patients with decompensated cirrhosis using validated ICD-9- 

code based algorithms, with at least one or more of the following 

infections: bacteremia, skin/soft tissue infection, urinary 

tract infection, lower respiratory infection, Clostridium difficile 

infection, and spontaneous bacterial peritonitis. We adjusted 

for patient co-morbidities using the Deyo modification of the 

Charlson index. We determined predictors of severe sepsis 

using logistic regression models. Results: Of 130,088 hospitalized 

patients with decompensated cirrhosis, 44,680 (34.4%) 

had ≥1 infection. Among infected patients, 8,077 (18.1%) 

had severe sepsis. Infected patients comprised 27,137 whites 

(71.1%), 3,704 black (9.7%) and 7,306 Hispanics (19.1%). 

White patients were older (mean 59.7y, p


2085 

Neutrophil to Lymphocyte Ratio (NLR) Predicts Liver 

Related Death in Low MELD Cirrhotic Patients Awaiting 

Liver Transplant 

Avash Kalra 1 , Joel P. Wedd 2 , Kiran Bambha 1 , Jane Gralla 1 , Hugo 

R. Rosen 1 , Scott W. Biggins 1 ; 1 University of Colorado Denver, 

Aurora, CO; 2 Division of Digestive Diseases, Emory, Atlanta, GA 

MELD score has reduced accuracy in cirrhotic patients with 

MELD ≤20. The D’Amico 5 Stages of Cirrhosis model is a 

cumulative clinical staging tool and aids in identifying low 

MELD patients at higher risk for liver related death (Liver Transpl 

2014; 20:1193-1201). High (≥4, J Hepatol 2013; 58:58-64) 

Neutrophil to Lymphocyte Ratio (NLR) is associated with inflammatory 

states and may predict cirrhotic decompensation and 

low MELD death in cirrhosis. AIM: To evaluate the prognostic 

utility of high NLR (≥4) for liver related death among low MELD 

patients listed for liver transplant (LT), controlling for Cirrhosis 

Stage. METHODS: A nested case-control study was performed 

using adults (>17 yo) with cirrhosis awaiting LT from 2/2002- 

5/2011; Cases= liver related death and MELD≤20 within 90 

days of death; and Controls= alive for ≥90 days after LT listing. 

Controls were randomly chosen from all listed patients 

and matched (up to 3:1) to Cases by listing year, lab MELD, 

age, gender, and disease etiology. NLR, Cirrhosis Stage, Na, 

albumin, and hepatic encephalopathy (HE) were assessed at 

date of lowest MELD within 90 days of death for Cases and 

within 90 days after listing for Controls. Cirrhosis Stages are 

1=no varices or ascites, 2=varices, 3=variceal bleed, 4=ascites, 

5=ascites and variceal bleed. Conditional logistic regression 

was used to evaluate risk of liver related death. RESULTS: 

There were 41 Cases and 66 Controls; MELD scores were similar. 

Clinical states contributing to death in Cases were: sepsis 

49%, spontaneous bacterial peritonitis 15%, variceal bleeding 

24%, and hepatorenal syndrome 22%. NLR 25 th , 50 th , 75 th 

percentile cut offs were 1.9, 3.1, and 6.8. NLR was ≥4 in 

4/41 (59%) Cases and 14/66 (21%) Controls. Median (IQR) 

NLR for Cases and Controls was 5.0 (3.0-10.3) and 2.35 

(1.6-3.9). In univariate analysis, NLR (continuous, ≥1.9, ≥4, 

≥6.8), increasing Cirrhosis Stage (**categorical,1-5), variceal 

bleeding, large ascites, HE, Na, Albumin were significantly 

(p 30 ng/ml) had significantly (P = 0.009) higher daily 

exposure to all thresholds of lux intensities, as compared to 

patients with vitamin D deficiency. A seasonal comparison confirmed 

significantly higher lux accrued in spring and summer (P 

< 0.0001). Median dietary vitamin D intake (1.48 mcg/day, 

0.26 - 12.12) was lower than typical intakes in the general 

population (2 - 4 mcg). We observed no correlations between 

light exposure and BMI or physical activity, neither between 

body fat mass and serum vitamin D levels. Light intensity was 

the only independent predictor of serum vitamin D level in multivariate 

regression analysis (P = 0.004). Serum vitamin D levels 

were highest in patients with > 30 min exposure at > 1000 

lux/day (P = 0.002). Conclusions: In CLD patients, serum vitamin 

D levels positively correlate with light exposure but not with 

dietary vitamin D intake. The contribution of vitamin D from diet 

is minimal and below the recently revised recommendations for 

vitamin D intake. In contrast, our findings highlight the need 

for combined interventions to treat vitamin D deficiency and 

may guide specific recommendations for sunlight exposure in 

patients with CLD. 

Disclosures: 

The following authors have nothing to disclose: Elisabeth Nick, Ralf Kaiser, Frank 

Lammert, Caroline S. Stokes


2087 

Polymorphisms in the interleukin (IL)-1 gene cluster are 

associated with multiple markers of systemic inflammation 

in patients with Acute-on-Chronic Liver Failure 

(ACLF) 

Jose Alcaraz-Quiles 1 , Esther Titos 4 , Cristina Lopez-Vicario 1 , 

Bibiana Rius 1 , Aritz Lopategi 1 , Mireia Casulleras 1 , Veronica 

Garcia-Alonso 1 , Andrea De Gottardi 5 , Mauro Bernardi 3 , Ivo Graziadei 

6 , Henning Gronbaek 7 , Pere Gines 8,4 , Vicente Arroyo 8,2 , 

Joan Clària 1,2 ; 1 Department of Biochemistry and Molecular Genetics, 

Hospital Clínic-University of Barcelona, Bareclona, Spain; 

2 University of Barcelona, Barcelona, Spain; 3 EASL-CLIF Consortium, 

Barcelona, Spain; 4 CIBERehd, Barcelona, Spain; 5 University 

of Berne, Berne, Switzerland; 6 Innsbruck Medical University, 

Innsbruck, Austria; 7 Aarhus University Hospital, Aarhus, Denmark; 

8 Liver Unit, Hospital Clinic, Barcelona, Spain 

Background and aim: Acute-on-Chronic liver failure (ACLF) is 

an increasingly recognized entity encompassing multiorgan 

failure in patients with cirrhosis. Recent findings suggest that 

immune dysregulation predisposes decompensated cirrhotic 

patients to a cycle of adverse events culminating in uncontrolled 

systemic inflammation and ACLF progression. Although 

the exact mechanisms leading to inflammation-driven ACLF 

remain to be elucidated, systemic inflammation is influenced by 

multiple genes regulating the innate immune response. In this 

study, we screened a set of six inflammation-related genes in 

a population-based study of patients with decompensated liver 

cirrhosis. Patients and Methods: Two hundred seventy-nine cirrhotic 

patients (178 with ACLF and 101 without ACLF) from the 

CANONIC study of the CLIF consortium were genotyped for 

SNP polymorphisms in genes coding for IL-1beta (rs1143623, 

G/C), IL-1 receptor antagonist (IL-1Ra) (rs4251961, T/C), 

SOCS3 (rs4969170, G/A), NOD2 (rs3135500, G/A), 

CMKLR1 (rs1878022, T/C) and IL-10 (rs1800871, G/A) by 

allellic discrimination TaqMan assays. Serum cytokine levels 

were measured by fluorescent bead-based (Luminex) immunoassays. 

Results: Among the six SNPs analyzed, we identified 

two polymorphisms belonging to the IL-1 gene cluster (IL-1beta, 

rs1143623 and IL-1Ra, rs425196) in strong association with 

ACLF. Homozygous C carriers in the rs1143623 SNP showed 

lower serum levels of IL-1β in parallel with reduced markers 

of systemic inflammation (i.e. IL-1alpha, IL-6 and TNF-alpha). 

Also, heterozygous TC carriers of the rs425196 SNP had 

lower serum levels of IL-1beta, IL-1alpha, IL-6, TNF-alpha and 

IL-8. Under different inheritance models, both genotypes were 

protective factors for presence of ACLF (rs1143623; OR: 0.34, 

p


2089 

Adding C-reactive Protein and Procalcitonin to the MELD 

Score Improves Mortality Prediction in Patients Admitted 

with Complications of Cirrhosis 

Sakkarin Chirapongsathorn 1,2 , Worawan Bunraksa 2 , Dollapas 

Punpanich 3 , Amnart Chaiprasert 4 , Patrick S. Kamath 1 ; 1 Gastroenterology 

and Hepatology, Mayo Clinic, Rochester, Rochester, 

MN; 2 Gastroenterology, Phramongkutklao Hospital and College 

of Medicine, Royal Thai Army, Bangkok, Thailand; 3 Biomedical 

Research and Development Center, Phramongkutklao Hospital and 

College of Medicine, Royal Thai Army, Bangkok, Thailand; 4 Nephology, 

Phramongkutklao Hospital and College of Medicine, Royal 

Thai Army, Bangkok, Thailand 

Background: MELD score has been shown to be the best predictor 

of mortality prediction in cirrhosis. Serum C-reactive protein 

(CRP) and procalcitonin (PCT) predict the risk of death in critical 

illness patients and also help to predict short-term mortality 

in cirrhosis. A model adding CRP and procalcitonin to the 

MELD score may increase accuracy for mortality prediction in 

patients admitted with complications of cirrhosis. Methods: A 

prospective cohort study was carried out in consecutive cirrhotic 

patients admitted with complications of cirrhosis during September 

2012 to December 2013 at Phramongkutklao Hospital, 

Thailand. All patients had venous CRP, PCT and laboratory 

values for MELD score calculation measured within 48 hours 

of admission. Cox regression analysis and ROC curve were 

used to predict mortality. The MELD-CRP score was externally 

validated in 818 eligible patients from Mayo Clinic, Rochester. 

Results: A cohort of 223 patients with cirrhosis was admitted 

during the study period. Seventy-one patients were eligible for 

analysis. MELD score was predictive of 90-day mortality (OR 

1.19 (95%CI; 1.09-1.32); AUROC of 0.81). Adding CRP or/ 

and PCT to the MELD score improved the predictive of 90-day 

mortality: MELD-CRP OR 2.71 (95% CI; 1.66-4.99); MELD-PCT 

OR 2.72 (95% CI; 1.66-4.99); MELD-CRP-PCT OR 2.71 (95% 

CI; 1.67-4.92). AUROC for MELD, MELD-CRP, MELD-PCT, and 

MELD-CRP-PCT were 0.81, 0.83, 0.84, and 0.85 respectively. 

Adding CRP or/and PCT to the MELD score also improved 

30-day mortality prediction. Similar results of MELD-CRP score 

were obtained from the Mayo Clinic external validation cohort. 

MELD-CRP score improved the predictive of 30-day mortality 

(OR 1.49 (95% CI; 1.28-1.73); AUROC 0.71) and 90-day 

mortality (OR 1.43 (95% CI; 1.27-1.62); AUROC 0.69) compared 

to MELD score. Conclusion: The MELD-CRP, MELD-PCT 

and MELD-CRP-PCT scores are superior to the MELD score in 

predicting mortality in cirrhotic patients admitted with complications. 


Worawan Bunraksa, Dollapas Punpanich, Amnart Chaiprasert 

2090 

Presepsin as a new biomarker for old expectations in 

the diagnosis and prognosis of bacterial infection in 

cirrhosis 

Maria Papp 1 , Tamas Tornai 1 , David Tornai 2 , Zsuzsanna Vitalis 1 , 

Istvan Tornai 1 , Peter Antal-Szalmas 2 ; 1 Department of Gastroenterology, 

University of Debrecen, Faculty of Medicine, Debrecen, 

Hungary; 2 Department of Laboratory Medicine, University of 

Debrecen, Faculty of Medicine, Debrecen, Hungary 

Background&Aims: Bacterial infections are frequent complications 

in cirrhosis with significant mortality. Early diagnosis is 

essential but still a diagnostic challenge from both the clinical 

and the laboratory part. The aim of this study is to evaluate 

and compare the diagnostic and prognostic value of presepsin 

plasma levels with CRP and PCT in bacterial infections of 

patients with cirrhosis. Methods: A total of 216 patients with 

cirrhosis (54.4% males, age: 57.6±10.3 years and median 

MELD score: 13 [95% CI: 10-17]) were consecutively enrolled. 

At admission enrollment presence of bacterial infection were 

assessed on the basis of conventional criteria, liver-oriented 

scores were calculated and plasma presepsin, CRP and PCT 

levels were measured. A short-term follow-up study was conducted 

to assess the development of organ system failure(s) and 

28-day mortality associated to bacterial infections. Results: Bacterial 

infection was found in 75 (34.7%) patients. Plasma presepsin 

levels were significantly higher in patients with infection as 

compared to those without (1002 pg/mL [575-2149] vs. 477 

[332-680] pg/mL, p0.5 pg/ml (OR: 

9.10, p=0.006) or CRP >40 mg/l (OR: 4.03, p=0.039) but 

not presepsin level were independet risk factor for 28-day mortality. 

Conclusions: Presepsin is a valuable new biomarker for 

defining severity of infections in cirrhosis proving same efficacy 

as PCT. However, for the prediction of short-term mortality, 

liver-oriented scores and admission level of conventional APP 

proteins, particularly PCT are the appropriate tools. 

Disclosures: 

Istvan Tornai - Advisory Committees or Review Panels: Roche, AbbVie, BMS, 

Janssen-Cilag 

The following authors have nothing to disclose: Maria Papp, Tamas Tornai, 

David Tornai, Zsuzsanna Vitalis, Peter Antal-Szalmas 

Disclosures: 

Patrick S. Kamath - Advisory Committees or Review Panels: Sequana Medical


2091 

The relationship of fecal bacterial isolates to systemic 

infections in cirrhotic patients 

Mary K. Rude 1 , Jimmy Ma 1 , Suki Chandrasekaran 2 , Phillip Tarr 2 , 

Jaquelyn Fleckenstein 1 ; 1 Gastroenterology, Washington University, 

Saint Louis, MO; 2 Pediatrics, Washington University, St. Louis, MO 

Background: Infections in patients with cirrhosis increase mortality 

4-fold, with the most frequent infections being spontaneous 

bacterial peritonitis and urinary infections. Bacterial 

translocation from the gut in cirrhosis is postulated to play an 

etiologic role in developing infection. However, no strategies 

exist to proactively risk-stratify which patients with cirrhosis will 

develop systemic infection. This study sought to determine if the 

pathogens can be isolated from stool prior to the onset or during 

systemic infections. Methods: Twenty-six adults with cirrhosis, 

hospitalized for a variety of indications, were prospectively 

enrolled and stool samples were obtained approximately every 

48 hours. Patients received routine medical care, and if there 

was clinical evidence of an infection, appropriate cultures were 

obtained. Fecal samples collected prior to the infectious event 

were used to recover the pathogens that subsequently invaded 

their bloodstreams and/or urine. We then sought to recover the 

same organisms in stool samples in enrolled patients who did 

not have evidence of a systemic infection. Culture-based techniques 

were used to recover stool bacteria that provisionally 

matched the bloodstream and/or urine organisms. MALDI-TOF 

was then used to confirm identity between fecal and systemic 

isolates. Results: Of the 26 subjects enrolled, 10 patients developed 

extraintestinal infections. Of these 10 patients, 3 were 

infected with Klebsiella pneumoniae, and 2 were infected with 

Staphylococcus aureus. K. pneumoniae was cultured from the 

stool and confirmed with MALDI-TOF in all 3 patients (6 stool 

samples) who subsequently developed K. pneumoniae infection. 

K. pneumoniae was recovered from only 7 of the 23 

patients (9 of 50 stool samples) who did not develop infection 

with K. pneumoniae (Fisher’s exact test, p


of NC, and 2.4% of CHF patients. Sepsis rose from 2.2% to 

5.7% (159% increase) in cirrhotic patients and by 2010 was 

greater than NC (5.3%) or CHF (3.1%) patients. 35.8% of 

cirrhotic patients with sepsis died compared to 17.9% of NC 

and 16.4 % of CHF patients (p


cirrhosis (M/F: 12/5, age: median 59.2 years (49-65), etiology 

of liver disease: HBV±HDV: 23.5%, HCV: 17.7%, alcohol: 

41.2%, other: 17.6%, Child-Pugh score: median 7.5 (5-12), 

MELD: median 10.5 (7-21), ascites (%): 3 (17.7) attending 

the outpatient clinics were included. None had hepatocellular 

carcinoma. Indications for PPIs administration were esophagitis 

(35%), peptic ulcer (59%) and others (6%). Bacterial DNA 

in serum and the levels of endotoxin, LBP, TGF-β, IL-1β, IL-6, 

IL-8, IL-12, IL-10, TNF-α and NOx were assessed before (time 

1) and at the end of treatment with PPIs (time 2). Bacterial 

DNA was amplified by PCR using universal 16SrRNA primers, 

endotoxin, LBP and TGF-β were measured by ELISA, the other 

cytokines by a Cytometric Bead Array method, and NO levels 

determined with a nitric oxide quantitation kit. Wilcoxon 

Signed Rank Test was used to evaluate significant differences 

in the parameters assayed before and after PPIs administration. 

Results: None of the patients developed an infection during the 

study period. Spearman’s correlation analysis demonstrated 

a correlation between baseline LBP and MELD score (r=0.63, 

p=0.02). Bacterial DNA was not detected before or after the 

treatment. Time 1: Median endotoxin: 3.19 (1.03-4.33), LBP: 

4.08 (3.23-12.99) μg/ml, NOx: 9.04 (2.44-15.72) μΜ, 

TGF-β: 4.85 (2.43-16.64) ng/ml, TNF-α: 3.05 (0-8.26), IL-1: 

4.7 (0-11.95), IL-6: 4.37 (0-16.09), IL-12: 7.69 (0-25.88), 

IL-10: 6.56 (0-27.87), IL-8: 103.4 (0-281.4) pg/ml. Time 2: 

Median endotoxin: 2.76 (1.49-86.55), LBP: 4.65 (0-17.53), 

NOx: 7.36 (0.56-23.08), TGF-β: 3.49 (1.54-15.94), TNFα: 

5.4 (0-60.98), IL-1: 8.57 (0-184.8), IL-6: 10.29 (0-31.5), 

IL-12: 15.96 (0-159), IL-10: 5.16 (0-34.8), IL-8: 64.45 (18.99- 

2892.4). No significant differences were observed between 

the concentrations of all measured indices between times 1 and 

2 (p>0.05). Subgroup analysis according to Child-Pugh stage 

yielded similar results. Conclusions: Short-term PPIs administration 

had no effect on serum bacterial DNA, bacterial products 

or cytokine concentrations in patients with liver cirrhosis. 

Disclosures: 

Christos K. Triantos - Speaking and Teaching: Bristol, Gilead 

The following authors have nothing to disclose: Maria Kalafateli, Panagiota Spadidea, 

Dimitrios Goukos, Efstratios Koutroumpakis, Stelios F. Assimakopoulos, 

Konstatinos Thomopoulos, Charalambos Gogos, Chrisoula Labropoulou-Karatza 

C, Athanasia Mouzaki, George Daikos, Vasiliki Nikolopoulou 

2096 

A prognostic model evaluating neutrophil-to-leucocyte 

ratio, organ failure and MELD scores as predictors of 

long-term survival in acute-on-chronic liver failure 

Danai Agiasotelli, Alexandra Alexopoulou, Larisa E. Vasilieva, 

Georgia Kalpakou, Spyros P. Dourakis; 2nd Dept of Medicine, 

Medical School University of Athens, Athens, Greece 

Purpose/Background: Acute-on chronic liver failure (ACLF) is 

defined as an acute deterioration of liver disease with high 

mortality in patients with cirrhosis. The time needed to reverse 

this condition and the factors affecting mortality after the early 

28-day-period have been evaluated. Methods: One-hundredninety-seven 

consecutive patients with cirrhosis were included. 

Patients were prospectively followed-up for 180 days. Results: 

ACLF was diagnosed in the 54.8% of patients. Infection was 

the most common precipitating event in patients with ACLF. 

Patients with ACLF had the worst prognosis compared to those 

without (log-rank P30 days & ≤75 days) and 

to 1.26 [(95% CI 0.53-3.00), P=0.609] during the third period 

of observation (>75 and


2098 

A novel mouse model for the study of pediatric Non-Alcoholic 


Veronica Marin 1 , Natalia Rosso 1 , Matteo Dal Ben 1 , Alan Raseni 2 , 

Cristina Degrassi 3 , Claudio Tiribelli 1,4 , Silvia Gazzin 1 ; 1 Fondazione 

Italiana Fegato, Trieste, Italy; 2 S.C.Laboratorio Analisi 

Cliniche, IRCCS Burlo Garofalo, Trieste, Italy; 3 Medical Research 

Institute, Trieste, Italy; 4 Department of Medical Science, University 

of Trieste, Trieste, Italy 

The increasing prevalence of pediatric NAFLD is considered 

a booming problem with worrying future outcome. Consistent 

pediatric models to mimic the clinical condition, as well as 

studies considering eventual gender differences, are still lacking. 

The aim of this study is to develop and characterize a 

Juvenile NAFLD model. Males (M) and females (F) C57BL/6 

mice, immediately after weaning were randomly assigned to 

control (CTRL) or high-fat high-carbohydrate diet (HFHCD). 

Animals had ad-libitum food for 16wks access. Body-weight, 

glycaemia, insulinemia, triglycerides, total cholesterol, HDL-C, 

LDL-C, ALT and liver histology were screened every 4wks and 

compared to CTRL. Soon after the 1 st week, HFHCD induced 

a significant bodyweight gain in both genders. Males, after 

4wks presented also hyperplasia of epididymal fat-pads and 

after week 12 th , a significant hepatomegaly. Males showed 

earlier alteration of glycemia, insulinemia, lipid profile and 

ALT (Table1). Interestingly, comparable body/blood alterations 

were observed in females only at the 16 th week. Liver histology 

showed in both genders a mixed macro-microvesicular steatosis 

increasing steadily after the 8 th week. Inflammatory cells foci 

were observed in males from the beginning and increased over 

the time. On the contrary, inflammation was absent in females. 

Surprisingly, both genders developed progressive fibrosis starting 

from the 8 th week and rising steadily over the time. This 

juvenile NAFLD model progresses faster than those previously 

reported in adults. A clear gender difference was found in the 

onset of the liver injury. Even if the final outcome was comparable 

between genders (fibrosis grade 2), males presented early 

signs of liver injury, whereas females did not. 

was January 2004 to March 2014. The total number of abnormal 

results for both caeruloplasmin and A1AT, defined as a 

value below the lower limit of normal, were assessed and subsequent 

diagnosis evaluated from medical records. The value 

for caeruloplasmin varied during the study, however the majority 

of samples were < 0.17 g/L. For A1AT deficiency the limit 

was < 0.9 g/L. Results During the study period caeruloplasmin 

was ordered on 2444 patients. The mean age of patients was 

46.4 years, 56.4% were male and 76 tests (3.1%) were abnormal. 

Plasma copper was performed on 41 patients (53.9%), 

however 36 (87.8%) were simultaneous with caeruloplasmin. 

Only 13 patients (17.1%) underwent 24-hour urinary copper 

measurement with 9 (69.2%) elevated. 21 patients (27.6%) 

had a liver biopsy, and 3 (14.3%) had hepatic copper quantification. 

Only 1 patient had WD confirmed. Other diagnoses 

included viral hepatitis (32.3%), alcohol (26.3%), drug induced 

liver injury (10.5%) and NAFLD (7.9%). The positive predictive 

value of a low caeruloplasmin level for WD was 1.4%. A1AT 

levels were requested for 3247 patients. The mean age of 

patients was 47 years, 55.9% were male and 88 tests (2.7%) 

were abnormal. Genotyping was performed on 55 patients 

(62.5%) with MZ (41.8%) most prevalent followed by MM 

(38.2%). Only 1 patient had ZZ genotype and was diagnosed 

with A1AT deficiency. Five patients had SZ genotype however 

only three had this noted in medical records. Of the 21 patients 

with MM genotype, 6 have been referred for further genotyping 

of rare genetic variants. 1 has shown MM procida 

. Other 

diagnoses included viral hepatitis (44.3%), NAFLD (18.2%), 

alcohol (11.4%) and drug induced liver injury (6.8%). The 

positive predictive value of a low A1AT level to detect ZZ genotype 

was 1.8%. Conclusion Caeruloplasmin and A1AT have 

very low detection rates when used as screening tests within the 

hepatology department of a tertiary hospital. The use of these 

investigations should be limited to patients in whom initial liver 

screen is negative and clinical suspicion remains. 

Disclosures: 

Leon A. Adams - Patent Held/Filed: Quest diagnostics 

The following authors have nothing to disclose: Tim Mitchell, John Beilby, Gary P. 

Jeffrey, George Garas, John Joseph, Ric Rossi, Gerry C. MacQuillan 

Disclosures: 

The following authors have nothing to disclose: Veronica Marin, Natalia Rosso, 

Matteo Dal Ben, Alan Raseni, Cristina Degrassi, Claudio Tiribelli, Silvia Gazzin 

2099 

The real world experience of screening investigations 

for Wilson’s disease and α1-antitrypsin deficiency 

within a tertiary liver unit – is it worthwhile? 

Tim Mitchell 1 , John Beilby 2 , Gary P. Jeffrey 1 , Leon A. Adams 1 , 

George Garas 1 , John Joseph 2 , Ric Rossi 2 , Gerry C. MacQuillan 1 ; 

1 Sir Charles Gairdner Hospital, Nedlands, WA, Australia; 2 Path- 

West, Nedlands, WA, Australia 

Introduction Wilson’s disease (WD) and α1-antitrypsin (A1AT) 

deficiency are rare but important causes of liver disease. The 

phenotypic presentation is varied and diagnosis can be difficult. 

Despite limitations as screening tests, caeruloplasmin 

and A1AT levels have been performed routinely at this center 

among patients presenting with liver disease. We reviewed the 

yield of these investigations to guide future ordering practices. 

Methods All serum caeruloplasmin and A1AT levels requested 

by the hepatology department at a tertiary hospital from the 

state reference laboratory were reviewed. The study period 

2100 

Relapse of Porphyria Cutanea Tarda After Achieving 

Remission With Phlebotomy or Low Dose Hydroxychloroquine 

Ashwani K. Singal 1 , Eric Gou 2 , Maira Rizwan 1 , Marisol Albuerne 2 , 

Csilla Kormos Hallberg 3 , Karl E. Anderson 3,2 ; 1 Gastroenterology 

and Hepatology, UAB, Birmingham, AL; 2 Internal Medicine, 

UTMB, Galveston, TX; 3 Preventive Medicine, UTMB, Galveston, TX 

Background: Porphyria cutanea tarda (PCT) is due to inhibition 

of hepatic uroporphyrinogen decarboxylase (UROD) and is 

effectively treated by phlebotomy (to serum ferritin of


(38%) patients relapsed biochemically (30% of 27 after phlebotomy 

and 50% of 18 after HCQ; Log Rank P=0.14). Relapse 

rates at 1, 3, and 5 years after phlebotomy or HCQ were 

8% vs. 24%, 30% vs. 30%, and 44% vs. 50% respectively. 

Relapse rates after these treatments did not differ by initial 

age, sex, race, alcohol use, smoking, HFE (hemochromatosis) 

genotypes C282Y/C282Y or C282Y/H63D, estrogen use, 

serum ferritin, or time to remission. Compared to non-relapsers, 

relapsing patients had higher pretreatment plasma porphyrin 

concentrations (14±20 vs.6±5 mcg/dL; P=0.046) and a higher 

proportion with HCV (94% vs. 71%, P=0.055). All remained 

abstinent from alcohol during treatment. But resumption of alcohol 

use tended to be more common among relapsers compared 

to non-relapsers (67 vs. 43%, P=0.1). Pooled data on relapse 

rate from 14 published studies on 1073 PCT patients showed 

relapse rates of 13.5% per year with phlebotomy and 17.9% 

per year with low dose hydroxychloroquine or chloroquine. 

The end-point of low dose HCQ or chloroquine treatment in all 

of these previous studies was normalization of urinary porphyrins 

with variable cut-off to define normal levels. Conclusions: 

PCT relapses may be more frequent with more severe disease 

and after achieving remission with low-dose HCQ than with 

phlebotomy. Longer follow-up and a larger sample size are 

planned with the Porphyrias Consortium study. The optimal 

length and end-point of treatment with low dose hydroxychloroquine 

need to be better defined. 

Disclosures: 

Karl E. Anderson - Consulting: Mitsubishi Tanabe Pharma America, Inc. 

The following authors have nothing to disclose: Ashwani K. Singal, Eric Gou, 

Maira Rizwan, Marisol Albuerne, Csilla Kormos Hallberg 

2101 

Hepatocellular Carcinoma in Patients with Wilson’s Disease: 

Results of a Single Centre Registry of 262 Patients 

Harshad Devarbhavi, Mallikarjun Patil; Department of Gastroenterology, 

St. John’s Medical College Hospital, Bangalore, India 

Introduction: Although hepatocellular carcinoma (HCC) is a 

well recognized complication of cirrhosis of various etiologies, 

HCC is believed to be extremely rare in Wilson disease. 

Around 30 cases have been reported worldwide illustrating 

the rarity of this complication. We undertook this study 

to determine the frequency and characteristics of HCC in a 

cohort of 262 patients over 19 years Patients and methods: 

We reviewed the case records of patients with Wilson disease 

from the Wilson disease registry maintained from 1996 to 

2014. Diagnosis of Wilson disease was based on a score > 

4 developed by the International group at Leipzig (J Hepatol 

2012;56:671-85). Patients were treated with D-penicillamine 

with or without Zinc Sulfate. Patients were regularly followed 

up at 3-6 monthly intervals. Patients with HCC were initially 

assessed by ultrasonography of abdomen followed by a contrast 

enhance CT scan. Results: We identified 3 cases of HCC 

among 262 patients with Wilson disease. All 3 had cirrhosis 

with portal hypertension at the time of diagnosis as evidenced 

by esophageal varices and ascites. The characteristics of these 

patients are depicted in Table. All three had multicentric HCC 

and presented with resistant complications of portal hypertension 

such as variceal bleeding, resistant ascites and one with 

hemoperitoneum. Given the advanced nature of HCC at presentation 

only supportive treatment could be given. Conclusions: 

The incidence of HCC in a large cohort of 262 patients 

is 1.1%. Although low, patients with WD are also at risk for 

HCC. HCC appears to be aggressive in our cohort of Wilson 

disease. Patients with Wilson disease need to undergo regular 

screening for early detection of HCC similar to other etiologies. 

Those who are insufficiently chelated appear to be more at risk 

for HCC. 

Characteristics of three patients with HCC 

Disclosures: 

The following authors have nothing to disclose: Harshad Devarbhavi, Mallikarjun 

Patil 

2102 

Congenital glycosylation defects mimicking Wilsons disease 

in patients with unexplained elevated aminotransferases 

and low ceruloplasmin 

Jos Jansen 1,2 , Joost Drenth 1 , Dirk Lefeber 2 , Monique van Scherpenzeel 

2 ; 1 Gastroenterology & Hepatology, Radboud University 

Medical Center Nijmegen, Utrecht, Netherlands; 2 Translational 

Metabolic Laboraty, glycosylation disorders, Radboud University 

Medical Center Nijmegen, Nijmegen, Netherlands 

Introduction Congenital disorders of glycosylation (CDGs) are 

a heterogenous group of autosomal recessive metabolic disorders 

with abnormal glycosylation as a hallmark. GI tract and 

liver symptoms are frequent but often secondary. Here we show 

that mutations in two uncharacterized genes are the cause 

for a primarily Wilson disease-like liver phenotype in twelve 

patients with unexplained glycosylation defects. Methods & 

Results These twelve patients (gene A: 5 pt., gene B: 8 pt.) 

showed an overlapping phenotype with elevated aminotransferases, 

high bone-derived alkaline phosphatase, hypercholesterolemia 

and elevated LDL-C. Further hepatic analysis showed 

low serum ceruloplasmin levels and liver biopsy indicated fibrosis, 

cirrhosis, steatosis and mild hepatic copper accumulation 

for some patients. Despite similarities, differences between the 

two gene defects were noticed. Gene A patients had a milder 

phenotype with slightly elevated aminotransferases and hypercholesterolemia. 

In contrast, gene B patients developed hepatosplenomegaly, 

resembling a glycogen or lysosomal storage 

disease, and end stage liver disease was seen for four patients. 

Additionally, for gene B, neurological symptoms were seen. 

Disease gene identification was done by functional analysis 

of exome sequencing. Based on yeast glycosylation models 

and BLAST searches two uncharacterized human proteins were 

identified. Subsequently analysis of raw exome sequencing 

data indicated incomplete coverage of gene A in a family 

where previous linkage and standard exome sequencing filtering 

was unsuccessful. Additional screening revealed six families 

with missense mutations, all segregating via an autosomal 

recessive inheritance. We analyzed intact serum transferrin 

via mass spectrometry for abnormal glycosylation and found 

a specific Golgi homeostasis defect profile. Additionally, we 

incubated patient fibroblasts with metabolic labeled sialic acid. 

Upon incorporation, the sialic acid gives a fluorescent signal. 

We show that fluorescence is reduced by app. 50% in tested 

patients. We transiently transfected HeLa cells with V5-tagged 

gene constructs and showed via immunofluorescence that 

both proteins are located in the ER-to-Golgi region. Together


with lipid vacuolization and dilated ER and Golgi on electron 

microscopy, these data point towards an effect on Golgi 

homeostasis for both defects. How this influences copper and 

lipid metabolism is still unknown. Conclusion We identified two 

new genes associated with unexplained elevated aminotransferases, 

low ceruloplasmin and hypercholesterolemia in twelve 

patients with abnormal glycosylation and suggest screening for 

glycosylation defects in these patients. 

Disclosures: 

The following authors have nothing to disclose: Jos Jansen, Joost Drenth, Dirk 

Lefeber, Monique van Scherpenzeel 

2103 

Functional analysis and drug response to zinc and 

D-penicillamine in stable ATP7B mutant hepatic cell lines 

Gursimran Chandhok, Vanessa Sauer, Christoph Niemietz, Andree 

Zibert, Hartmut H. Schmidt; Klinik für Transplantationsmedizin, Universitätsklinikum 

Münster, Münster, Germany 

Objective: To assess in vitro functional analysis and drug 

response to zinc (Zn) and D-penicillamine (DPA) treatment of 

most common disease-causing ATP7B mutations observed in 

Wilson disease (WD) patients. Background: WD is a rare autosomal 

recessive disorder caused due to mutations in the copper 

transporter ATP7B. More than 600 disease-causing mutations 

are described. If not treated, WD is fatal. Commonly used 

drugs to reverse Cu toxicity in WD include direct chelation by 

DPA or metallothionein induction using Zn salts. The impact of 

these drugs in rescuing hepatocytes carrying individual mutations 

is unknown. Methods: Frequent homozygous mutations in 

Europe/US, Asia, and India were selected for functional analysis. 

An ATP7B knockout hepatic cell line previously established 

by our group (PLoS ONE, 9: e98809, 2014) was used to generate 

stable mutant cell lines. ATP7B gene and protein expression 

were determined for all mutant cell lines. Cu induced cell 

survival, apoptosis, and intracellular trafficking were studied. 

Cell viability against Cu toxicity was determined on treatment 

with Zn, DPA, or both. Results: ATP7B mRNA was similar 

whereas protein expression varied among the different mutant 

cell lines. Co-localization studies with late endosome lysosome 

marker lamp2 suggested impaired trafficking in presence or 

absence of Cu. All mutant cell lines showed different grades of 

ATP7B activity against Cu induced toxicity and apoptosis with 

p.H1069Q displaying moderate ATP7B activity. On treatment 

with Zn, DPA, or both most cell lines showed a characteristic 

response to treatment. DPA was more effective than Zn in rescuing 

cells against Cu toxicity. Only combination with Zn and 

DPA treatment was able to rescue the mutant cells similar to 

wild-type. Conclusions: The study provided insights into genotype-phenotype 

correlation in WD. Different ATP7B mutations 

showed varying sensitivity to Cu with high/moderate and low 

ATP7B activity. Individual ATP7B mutations showed a genotype-specific 

response to treatment. Combination treatment with 

Zn and DPA showed maximal survival of hepatic cells. Our 

data suggest that prognosis of WD patients may benefit from 

knowledge of mutation-specific in vitro functional analysis and 

use of Zn/DPA combination therapy. 

Disclosures: 

The following authors have nothing to disclose: Gursimran Chandhok, Vanessa 

Sauer, Christoph Niemietz, Andree Zibert, Hartmut H. Schmidt 

2104 

Cardiac MRI for Assessment of Iron Overload in Cirrhotic 

Patients Evaluated for Liver Transplantation 

R Todd Frederick 1 , Christopher R. Kagay 2 , Peter Hui 3 , Richard 

E. Shaw 3 , Robert W. Osorio 1 ; 1 Department of Transplantation, 

California Pacific Medical Center, San Francisco, CA; 2 Radiology, 

California Pacific Medical Center, San Francisco, CA; 3 Cardiology, 

California Pacific Medical Center, San Francisco, CA 

Background and Aims: Pathologic iron overload of the liver 

is often associated with cardiac iron overload and can have 

deleterious effects on cardiac function and compromise postliver 

transplant (LT) outcomes. We reviewed the experience 

at our center using cardiac MRI to estimate cardiac iron content 

in the evaluation of cirrhotic patients for LT. Methods: All 

patients undergoing evaluation for LT with moderate to severe 

hepatic iron overload (>2+ iron stain and/or >4000mcg/gm 

by liver biopsy or liver MRI) required cardiac MRI with calculation 

of T2* signal decay (GE StarMap). A minimum of 

3 regions of interest were selected and the mean T2* was 

determined (msec). Endomyocardial biopsy (EMBx) was performed 

for patients with suspected iron overload meeting all 

other LT criteria. EMBx was graded as normal (no iron), “borderline” 

(trace or 1+), “moderate” (2-3+), or “severe” (>3-4+). 

Comparison of T2* with iron content on EMBx was assessed 

using ANOVA. Results: 28 patients with hepatic iron overload 

underwent cardiac MRI between 6/2012 - 4/2015. Mean 

age was 54 (26-66), 71% male, 68% Caucasian. Etiology of 

liver disease included alcohol (9), HCV (5), HCV/alcohol (4), 

hereditary hemochromatosis (4), cryptogenic/biliary (3), sickle 

cell (1), HBV (1), and NASH (1). Mean transferrin saturation 

was 86.1% and mean ferritin 1600 ng/mL. The mean T2* 

was 37.2 msec (range 11.75-88.5, normal > 40). 15 patients 

underwent EMBx. 7 were considered “positive” for iron (3 

severe and 4 moderate), 4 were borderline, and 4 were negative. 

All 7 patients with “positive” EMBx were declined for LT 

at our center per protocol; 5 subsequently died (one following 

LT at another center), while 2 are awaiting LT at other centers. 

Of the 4 “borderline” patients, 1 underwent LT with complications, 

1 was declined for CAD (died), 1 died of sepsis, and 1 

remains listed. Of the 4 with negative cardiac iron, 1 received 

LT, 1 died of HCC, and 2 remain listed. The mean values of 

T2* were significantly different when compared across EMBx 

iron content groups (p=0.001). Using a T2* cutoff of 30msec 

provided 100% PPV and NPV for EMBx for “positive” vs. negative 

or “borderline” iron. Conclusions: Cardiac MRI estimation 

of myocardial iron utilizing T2* accurately identifies endomyocardial 

iron loading as identified by EMBx and may be useful 

in stratifying risk prior to liver transplantation. 

Comparison of T2* with EMBx iron (p= 0.001) 

Disclosures: 



The following authors have nothing to disclose: Christopher R. Kagay, Peter Hui, 

Richard E. Shaw, Robert W. Osorio


2105 

Genetic variants associated with liver disease in α-1-antitrypsin 

deficiency 

Richard J. Thompson 1 , Jeid Turquet 1 , Rosamond Nuamah 2 , Patrick 

J. McKiernan 3 , Robert Stockley 4 , Gerome Breen 5 , Nedim Hadzic 6 ; 

1 Institute of Liver Studies, King’s College London, London, United 

Kingdom; 2 BRC Genomics Facility, Guy’s and St Thomas’ NHS 

Foundation Trust, London, United Kingdom; 3 Liver Unit, Birmingham 

Children’s Hospital, Birmingham, United Kingdom; 4 Lung 

Immuno Biochemical Research Laboratory, University of Birmingham, 

Birmingham, United Kingdom; 5 Institute of Psychiatry, King’s 

College London, London, United Kingdom; 6 Paediatric Liver, GI 

and Nutrition Centre, King’s College Hospital, London, United 

Kingdom 

Background Alpha-1-antitrysin is a major plasma serine protease 

inhibitor. It is encoded by SERPINA1. Several deficient 

alleles have been described. The Z allele (p.E342K) leads to 

protein polymerisation and intracellular retention. In homozygous 

form this allele is seen in patients with lung and liver 

disease; α-1-antitrypsin deficiency (A1ATD). The respiratory disease 

is highly penetrant, whereas neonatal cholestasis is seen 

in only 10-15% of Z homozygotes. The cause of the reduced 

penetrance seen in this liver phenotype is unexplained. Aim 

To identify genetic loci associated with neonatal presentation 

of A1ATD associated liver disease. Methods All patients were 

homozygous for p.E342K, the Z allele. One group consisted 

of 384 patients followed in an adult respiratory clinic. The 

second group consisted of 132 patients presenting with neonatal 

cholestasis. All were genotyped using Illumina Infinium 

HumanCoreExome arrays; interrogating >240,000 tagSNPs. 

Data was analysed using several software tools including 

GenomeStudio, PLINK and PRSice. Thus allowed single locus 

and polygenic associations to be identified. Results Differences 

in SNP allele frequencies between the 2 groups of patients 

were studied. Three separate loci with a lod score of >6 were 

identified. They are on chromosomes 1, 12 and 14. The latter 

is 10 Mb from the SERPINA1 gene. A large number of other 

loci have been subject to further analysis, looking for polygenic 

and pathway effects. Only 1 of the three loci lies within a gene. 

This in turn has not been linked to liver disease. Conclusions 

Although AIATD-associated liver disease is thought of as being 

genetic, the major locus (SERPINA1) shows markedly reduced 

penetrance. The fact that all Z alleles are inherited from a 

relatively recent ancestor means that all affected individuals 

are much more closely related than in nearly all similar studies. 

This in turn means that between phenotypically diverse 

groups, genetic associations can be identified using relatively 

small sample sizes. This proved to be the case and allowed the 

identification of 3 quite separate loci conferring susceptibility 

to liver disease. The mechanisms by which this risk is conferred 

remain to be identified, however this is the first step in the identification 

of the cause of A1ATD liver disease. 

Disclosures: 

Richard J. Thompson - Grant/Research Support: Shire; Speaking and Teaching: 

Shire 

Patrick J. McKiernan - Consulting: Alnylam Pharmaceuticals 

Nedim Hadzic - Consulting: Alnylam; Speaking and Teaching: Synageva 

The following authors have nothing to disclose: Jeid Turquet, Rosamond Nuamah, 

Robert Stockley, Gerome Breen 

2106 

Clinical features, biochemical measures and liver histology 

in Hereditary Hemochromatosis Argentinian 

patients with and without HFE gene mutations. 

Florencia Yamasato, Marcelo Castro, Alejandra Avagnina, Alejandra 

Vellicce, Alejandra Pernazza, Jorge Rey, Esteban González 

Ballerga, Juan A. Sorda, Jorge Daruich; Gastroenterologia, Hospital 

de Clínicas José de San Martín. Universidad de Buenos Aires, 

Buenos Aires, Argentina 

Introduction: Inclusion of HFE gene mutations, present in 

90-96% of Hereditary Hemochromatosis (HH) Anglo-Saxon 

patients (Pt), has been a great advance in diagnosis algorithm. 

However, the percentage of these mutations among HH Pt 

could be different in our population. Objectives: To describe 

in HH Pt, the differences in clinical and biochemical features, 

and the presence of cirrhosis among subjects with HFE gene 

mutations (G1) and those without them (G2). Materials and 

Methods: Cross sectional study. 277 Pt with HH diagnosis 

between 1989 and 2014 were included. Only Pt with studied 

HFE gene mutations were included. HH diagnosis was 

performed by internationally accepted criteria and compatible 

liver histology. HOMA, Diabetes Mellitus (DM), alcohol 

intake (>30g/d), articular and cardiac features, liver histology 

(cirrhosis) and non-alcoholic fatty liver disease (NAFLD) were 

investigated and compared between both groups. Statistical 

analysis: Descriptive statistics are displayed as Median values 

with 95% Confidence Intervals (CI). Mann Whitney method 

and logistic regressions were performed. P Values


2107 

Mutational analysis of ATP7B in Chinese Wilson disease 

patients 

Shan S. Peng, Lingxia QI, yuanyuan Cui, Wenli Kong, Linyuan 

Ma, Yu Pan, Junqi Niu, Rui Hua; Hepatology, The First Hospital, 

Jilin University, Changchun, China 

Wilson Disease (WD) is an inborn error of copper metabolism 

inherited in an autosomal recessive manner caused by 

the mutations in the P-type ATPase gene (ATP7B). In this study, 

we screen and detect the mutations of the ATP7B gene in unrelated 

Chinese WD patients.A total of 62 individuals from ten 

provinces of china with WD were recruited. Of them, 41 were 

males and 21 were females, and their onset ages were from 5 

to 61 years with a median age of 26 years. The full length of 

ATP7B gene of the patients was sequenced and aligned to the 

referred ATP7B gene sequence. The results suggested that all 

the patents carried with mutations and 48 different mutations 

were identified, of which 35 were missense, one was nonsense, 

two were splicing, and 10 were frameshift (five insertion 

and five deletion). Among these mutations, c.2333G>T 

(32.3%), c.2975C>T (11.5%), and c.3443T>C (10.4%) were 

the most prevalent mutants and c.2310C>G always linked with 

c.2333G>T. The eighth, 11 th , and 18 th exons carried more 

mutations (6/48, 5/48, and 5/48, respectively). After comparing 

with the mutations reported previously, 19 out of the 48 

mutations were novel made up of all the splicing and frameshift 

mutations and 10 of 35 missense mutations. Two popular algorithms 

were used to predict the effects of the novel mutations 

on ATP7B function and most of the mutations were unfavorable 

(18/21). Our study will broaden our knowledge about ATP7B 

mutations in WD patients in China and further analysis need to 

examine the relationship between these mutations and clinical 

characteristics of WD. 

Disclosures: 

The following authors have nothing to disclose: Shan S. Peng, Lingxia QI, 

yuanyuan Cui, Wenli Kong, Linyuan Ma, Yu Pan, Junqi Niu, Rui Hua 

2108 

UGT1A regulation by a member of the proto-oncogenic 

miR-106b-25 cluster: hsa-miR-25 

Sandra Kalthoff, Anja Winkler, Christian P. Strassburg; Department 

of Internal Medicine, University Hospital Bonn, Bonn, Germany 

Introduction: UDP-glucuronosyltransferases (UGTs) catalyze 

the detoxification of xenobiotics including a variety of carcinogens. 

Therefore, UGTs play an essential role in defence 

mechanisms protecting the organism against the development 

of cancer. Recent research has shown that deregulation of 

miRNAs can contribute to cancer development. In hepatocellular 

carcinoma (HCC) tissues, hsa-miR-25, for example, is 

overexpressed and associated with a poor prognosis in HCC 

patients. Aim of this study was to examine the influence of hsamiR-25 

on UGT1A-expression. Methods: For luciferase assays, 

reporter gene constructs containing different UGT1A promoters 

(UGT1A1, UGT1A3, UGT1A4, UGT1A7, UGT1A9), 

combined with luciferase sequence and the UGT1A 3’untranslated 

region (UTR) sequence were generated. Every promoter 

was combined with either the UGT1A 3’UTR-wild type(WT) 

sequence or the UGT1A 3’UTR-SNP sequence (containing the 

3 SNPs rs10929303, rs1042640 and rs8330). The constructs 

were transfected together with 5 nM hsa-miR-25 into HepG2 

cells. For RNA quantification HepG2 cells were treated with 5 

nM hsa-miR-25 and isolated RNA was analyzed by TaqMan 

PCR. Results: Transfection of hsa-miR-25 led to a significant 

decrease in luciferase expression of the UGT1A1-, UGT1A3-, 

UGT1A4- UGT1A7 and UGT1A9 reporter gene constructs (0.4- 

0.5 fold). Interestingly, transfection of the construct containing 

the UGT1A7 promoter combined with the UGT1A 3’UTR-SNP 

sequence, led to a further reduction of luciferase expression 

compared to the UGT1A 3 ‘UTR-WT sequence (SNP: 0.21 fold 

compared to UGT1A 3’UTR-WT 0.42 fold). RNA analysis of 

HepG2 cells revealed that mRNA expression of UGT1A1 (0.47 

fold), UGT1A3 (0.49 fold), UGT1A4 (0.68 fold), UGT1A6 

(0.6 fold), UGT1A7 (0.38 fold) and UGT1A9 (0.36 fold) were 

significantly reduced by treatment with 5 nM hsa-miR-25. Conclusions: 

This is the first demonstration of UGT1A regulation 

by hsa-miR-25. All UGT1A genes were significantly downregulated 

by treatment with 5 nM hsa-miR-25 on mRNA level as 

well in reporter gene assays. The presence of 3 SNPs in the 

UGT1A 3’untranslated region combined with the UGT1A7 

promoter led to a further reduction of luciferase expression 

indicating a suppressive effect of theses SNPs on the activity 

of the major carcinogen detoxifying UGT1A7 enzyme in the 

presence of overexpressed hsa-miR-25. The elucidated significant 

reduction of UGT1A expression and the accompanying 

affected detoxification capacity mediated by hsa-miR-25 may 

represent a possible explanation for the potential association 

between hsa-miR-25 and HCC development. 

Disclosures: 



The following authors have nothing to disclose: Sandra Kalthoff, Anja Winkler 

2109 

Wilson disease: gestational methyl group supplementation 

affects hepatic oxidative phosphorylation and 

movement disorder pathways in fetal mouse liver 

Valentina Medici, Noreene Shibata, Charles H. Halsted, Janine M. 

LaSalle; University of California Davis, Sacramento, CA 

Background & Hypothesis. Wilson Disease (WD) is a condition 

with highly variable phenotypic presentation, including 

hepatic and neurological involvement. Previously we showed 

that methionine metabolism, central in the regulation of methylation 

reactions, is altered in the tx-j mouse model of WD and 

therefore may influence the expression of genes involved in 

liver damage. Since maternal diet is known to affect fetal gene 

transcript levels through epigenetic mechanisms, we hypothesize 

the phenotype of WD is influenced by maternal methylation 

status. Methods. The tx-j mouse model of WD and wildtype 

C3H female mice were started on choline-supplemented (choline 

36 mmol/Kg of diet) and control diets (choline 8 mmol/ 

Kg of diet) 2 weeks before mating and through pregnancy 

to embryonic day 17. Livers of 6-11 embryos/dam were collected 

for RNA-sequencing. Pathway analysis was performed 

by DAVID software. An additional group of offspring was 

euthanized at 24 weeks to confirm persistence of changes in 

gene expression in adult livers. Results. Comparing tx-j and 

wildtype fetal livers, pathway analysis revealed differences 

in transcript levels of nuclear encoded genes related to oxidative 

phosphorylation, which overlapped with genes related to 

neurological movement disorders, including Huntington’s and 

Parkinson’s disease, both conditions listed in the differential 

diagnosis of WD. Maternal choline supplementation was associated 

with maintenance of gene transcript levels at control 

group levels in fetal livers. Transcript levels of neurexin-1, central 

to cell adhesion in the nervous system, was up-regulated 

67-fold in fetal livers of tx-j mice compared to wildtype mice, 

but were maintained at control levels in offspring of choline 

supplemented dams. In 24-week old tx-j offspring mouse livers, 

we found persistent changes in transcript levels of nuclear


encoded genes related to oxidative phosphorylation, including 

succinate dehydrogenase A (Sdha), a major catalytic subunit 

of the succinate-ubiquinone oxidoreductase, a complex of the 

mitochondrial respiratory chain. Of note, maternal choline 

maintained control levels of gene transcript levels of Sdha. 

Conclusions. Fetal and 24 week old livers of a mouse model 

of WD are characterized by changes in transcript levels of 

genes related to the oxidative phosphorylation pathway which 

is also involved in neurological diseases and these findings 

were prevented by maternal choline supplementation. Since 

these findings were present in both fetal and 24 week old tx-j 

livers, they may influence WD phenotype. 

Disclosures: 

The following authors have nothing to disclose: Valentina Medici, Noreene Shibata, 

Charles H. Halsted, Janine M. LaSalle 

2110 

Association between Psoas Muscle FDG Uptake and 

Metabolic Syndrome by 18F-Fluorodeoxyglucose Positron 

Emission Tomography: May psoas muscle FDG 

uptake use as a biomarker for impaired metabolic 

health? 

Seung Min Lee 1 , Dae Won Jun 2 , Yong Kyun Cho 3 , Eun Chul Jang 4 , 

Joo Hee Kwak 2 ; 1 Department of Food and Nutrition, Sungshin 

Women’s University, Seoul, Korea (the Republic of); 2 Department 

of Internal Medicine, Hanyang University College of Medicine, 


Kangbuk Samsung Hospital, Sungkyunkwan University, School of 

Medicine, Seoul, Korea (the Republic of); 4 Department of Occupational 

and Environmental Medicine, Soonchunhyang University 

Cheonan Hospital, Cheonan, Korea (the Republic of) 

Purpose: To identify metabolic activity of several organs 

responsible for the glucose metabolism on 18 F-FDG PET/CT 

according to several metabolic phenotypes and to evaluate 

whether it could be used to predict the metabolic health impairment. 

Methods: We retrospectively analyzed the data from 

157 subjects who underwent 18 F-FDG PET/CT for health medical 

examination. Liver function test, total cholesterol, triglyceride, 

and fasting blood glucose (FBG) level as well as presence 

of metabolic syndrome (MS) were evaluated in these subjects. 

Fixed or flexible spherical volume of interest (VOIs) were used 

to evaluate maximal and/or mean standardized uptake value 

(SUV) to assess the metabolism of liver, pancreas, mesenteric 

visceral fat, psoas muscle, and abdominal subcutaneous fat on 

18 F-FDG PET/CT. Effect of SUVs in each organ on the clinical 

metabolic parameters as well as on the presence of MS were 

analyzed for statistical significance. Results: 52 subjects (33.1 

%) were obese with a body mass index (BMI) > 25 and forty 

subjects (40/157, 25%) had MS. SUVmax of psoas muscle, 

mesenteric visceral fat, and abdominal subcutaneous fat as 

well as SUVmean of liver were positively correlated with BMI, 

weight and FBG, after adjusted by FBG. SUVmax of psoas 

muscle, visceral fat, and pancreas were significantly higher in 

the subjects with obesity and in the subject with MS (p for trend 

< 0.05). Among them, SUVmax of psoas muscle was found 

to be only risk factor for the presence of MS independent to 

weight and FBG. With the cutoff value of 1.34, SUVmax of 

psoas muscle accurately predicted MS, central obesity, and 

hypertension (AUROC 0.779, 0.810, and 0.646, respectively, 

p < 0.05). Conclusion: SUVmax of psoas muscle on the 18 F- 

FDG PET/CT is well associated with various clinical metabolic 

parameters and it could be used to predict the metabolic health 

impairment. Moreover, it could be valuable tool to sort metabolically 

healthy or abnormal especially in the obese subjects. 

Disclosures: 

The following authors have nothing to disclose: Seung Min Lee, Dae Won Jun, 

Yong Kyun Cho, Eun Chul Jang, Joo Hee Kwak 

2111 

Pitfalls in Erythrocyte Protoporphyrin Measurement for 

Diagnosis and Monitoring of Protoporphyrias 

Eric Gou 1 , Manisha Balwani 3 , D Montgomery Bissell 4 , Joseph 

R. Bloomer 5 , Herbert L. Bonkovsky 2 , Robert J. Desnick 3 , John D. 

Phillips 6 , Ashwani Singal 5 , Bruce M. Wang 4 , Hetanshi Naik 3 , 

Karl E. Anderson 1 ; 1 Preventive Medicine and Community Health, 

University of Texas Medical Branch, Galveston, TX; 2 Wake Forest 

University, Winston-Salem, NC; 3 Icahn School of Medicine at Mt. 

Sinai, New York, NY; 4 University of California San Francisdo, San 

Francisco, CA; 5 Universtiy of Alabama Birmingham, Birmingham, 

AL; 6 University of Utah, Salt Lake City, UT 

Erythropoietic protoporphyria (EPP) is due to the inherited 

deficiency of ferrochelatase, the enzyme that catalyzes the 

chelation of iron with protoporphyrin IX to complete heme biosynthesis. 

Increased erythrocyte and plasma protoporphyrin 

levels in EPP cause painful photosensitivity and impair quality 

of life. EPP is the third most common porphyria, the most common 

in children and its diagnosis is often delayed. Diagnosis 

of EPP requires: (1) a marked increase in total erythrocyte protoporphyrin 

(300-5,000 ug/dL erythrocytes) and (2) a predominance 

(85-100%) of metal-free protoporphyrin. X-linked 

protoporphyria (XLP) is less common and is due to gain of function 

mutations of the erythroid form of d-aminolevulinic acid 

synthase, the first enzyme in the heme biosynthetic pathway. 

XLP causes a similar increase in total erythrocyte protoporphyrin 

but with a somewhat lower fraction of metal-free protoporphyrin 

(50-85% of the total). The Porphyrias Consortium (PC) 

has enrolled more than 180 patients with EPP and XLP. Among 

the few patients with original laboratory reports of erythrocyte 

protoporphyrin levels, 15 appeared to have much higher levels 

(4.7 to 46.7 fold) when enrolled in studies at a PC Center. The 

diagnosis had been missed in 2 of these patients based on 

the earlier spurious results. All of the prior reports were from 

either Quest or LabCorp, which use only hematofluorometry for 

assessing erythrocyte protoporphyrin. The hematofluorometer 

was developed to screen for lead poisoning, and measures 

zinc protoporphyrin by front surface fluorescence using a drop 

of blood. The instrument displays two calculations based on 

assumed hematocrits each specified by OSHA and CDC, and 

variously reports results as “free” and “zinc” protoporphyrin 

(with different reference ranges), implying separate measurements 

of metal-free and zinc protoporphyrin. These reports are 

misleading and impair diagnosis and monitoring of patients 

with protoporphyria. This is concerning because reproducible 

methods are needed for diagnosis and longitudinal monitoring 

of erythrocyte protoporphyrin, as rising levels may predict 

severe hepatic complications requiring liver transplantation.


We suggest that laboratories offering erythrocyte protoporphyrin 

measurement should prioritize testing for EPP and XLP, 

which are essential for diagnosis and monitoring of these conditions, 

and less important for diagnosis of lead poisoning, iron 

deficiency and other erythrocyte disorders. Terms and abbreviations 

used in reporting results should be accurately defined. 

Disclosures: 

Joseph R. Bloomer - Advisory Committees or Review Panels: Recordati Rare 

Diseases; Consulting: Mitsubishi Tanabe Pharma; Grant/Research Support: Clinuvel, 

Inc., American Porphyria Foundation, NIH 5U54 DK083909, Alnylam 





Robert J. Desnick - Advisory Committees or Review Panels: Recordati Rare Diseases; 

Consulting: Alnylam Pharmaceuticals; Grant/Research Support: Alnylam 

Pharmaceuticals; Patent Held/Filed: Alnylam Pharmaceuticals; Stock Shareholder: 



The following authors have nothing to disclose: Eric Gou, Manisha Balwani, D 

Montgomery Bissell, John D. Phillips, Ashwani Singal, Bruce M. Wang, Hetanshi 

Naik 

2112 

Variation in erythrocyte and plasma protoporphyrin levels 

over time in protoporphyrias 

Eric Gou 1 , Cindy Weng 2 , John D. Phillips 2 , Tom Greene 2 , Manisha 

Balwani 4 , D Montgomery Bissell 6 , Joseph R. Bloomer 5 , Herbert L. 

Bonkovsky 3 , Robert J. Desnick 4 , Hetanshi Naik 4 , VM Sadagoparamanujam 

1 , Karl E. Anderson 1 ; 1 Preventive Medicine and Community 

Health, University of Texas Medical Branch, Galveston, TX; 

2 University of Utah, Salt Lake City, UT; 3 Wake Forest University, 

Winston-Salem, NC; 4 Icahn School of Medicine at Mt. Sinai, New 

York, NY; 5 University of Alabama, Birmingham, AL; 6 University of 

California, San Francisco, CA 

Background: Erythropoietic protoporphyria (EPP) and X-linked 

protoporphyria (XLP) cause painful, non-blistering cutaneous 

photosensitivity. EPP, the third most common porphyria and the 

most common in children, is due to decreased activity of ferrochelatase 

(FECH), which catalyzes the insertion of iron into protoporphyrin 

IX to complete heme synthesis. XLP is less common 

and is due to gain of function mutations of the erythroid form 

of d-aminolevulinic acid synthase, the first enzyme in the heme 

biosynthetic pathway. Hepatotoxic effects of excess protoporphyrin 

cause protoporphyric hepatopathy in less than 5% of 

patients. Hepatopathy impairs protoporphyrin excretion, which 

leads to further increases in circulating protoporphyrin levels 

and accelerated liver damage, often leading to liver transplantation. 

Normal variability in protoporphyrin levels over time has 

not been characterized in EPP and XLP, and whether threshold 

levels lead to hepatopathy is not known. Purpose: To compare 

porphyrin levels in EPP and XLP, and characterize expected 

variability of erythrocyte and plasma protoporphyrin levels in 

the absence of hepatopathy. Methods: Coefficients of variation 

(CV) were determined for levels of erythrocyte and plasma 

protoporphyrin repeated at least 4 times in 86 subjects with 

documented EPP or XLP. Patients with protoporphyric hepatopathy 

or elevated transaminases were excluded. Results: Total 

erythrocyte protoporphyrin was higher, on average, in XLP 

than in EPP, with considerable overlap. Total erythrocyte protoporphyrin 

levels were higher, on average, in males with XLP 

than females. Differences in erythrocyte protoporphyrin levels 

were lower between related than unrelated subjects with EPP or 

XLP. The ratio of metal-free to zinc protoporphyrin was higher 

in EPP, and was very stable over time. The CV for repeated 

values of total erythrocyte protoporphyrin averaged 19.59, 

and was not significantly different in EPP and XLP (19.06 and 

23.06 respectively). Plasma porphyrin levels were much lower, 

correlated only roughly with erythrocyte levels, and were much 

more variable over time. To date, none in this cohort has developed 

hepatopathy or sustained upward trends in protoporphyrin 

levels. Conclusion: Variation in erythrocyte protoporphyrin 

levels of up to ~25% may occur in EPP and XLP in the absence 

of liver disease. Studies in more patients over longer periods 

of time are in progress to see if greater increases or threshold 

levels of erythrocyte and plasma protoporphyrin precede the 

development of overt protoporphyric hepatopathy. 

Disclosures: 

Joseph R. Bloomer - Advisory Committees or Review Panels: Recordati Rare 

Diseases; Consulting: Mitsubishi Tanabe Pharma; Grant/Research Support: Clinuvel, 

Inc., American Porphyria Foundation, NIH 5U54 DK083909, Alnylam 





Robert J. Desnick - Advisory Committees or Review Panels: Recordati Rare Diseases; 

Consulting: Alnylam Pharmaceuticals; Grant/Research Support: Alnylam 

Pharmaceuticals; Patent Held/Filed: Alnylam Pharmaceuticals; Stock Shareholder: 



The following authors have nothing to disclose: Eric Gou, Cindy Weng, John D. 

Phillips, Tom Greene, Manisha Balwani, D Montgomery Bissell, Hetanshi Naik, 

VM Sadagoparamanujam 

2113 

A Nationwide, Population-based Epidemiology and 

Disease Burden of Wilson ’s disease in South Korea in 

2009-2013 

Joo Yeong Baeg 1 , Eun Sun Jang 1 , Mo-ran Ki 3 , Bo Hyun Kim 3 , 

Kyung-ah Kim 4 , Hwa Young Choi 2 , Sook-Hyang Jeong 1 ; 1 Internal 

Medicine, Seoul National University College of Medicine, 

Seongnam-Si,, Korea (the Republic of); 2 Department of Cancer 

Control and Policy, Graduate School of Cancer Science and Policy, 

National Cancer Center, Goyang, Korea (the Republic of); 

3 Center for Liver Cancer, National Cancer Center,, Goyang, 

Korea (the Republic of); 4 Departments of Internal Medicine, Inje 

University Ilsan Paik Hospital, Goyang, Korea (the Republic of) 

Background/Aims: Wilson’s disease (WD) is an inherited autosomal 

recessive disorder of copper metabolism in which copper 

accumulates in the organ, particularly in the liver and the central 

nervous system. As a rare disease, population-based epidemiology 

of WD is largely unknown. This study aimed to investigate 

the nationwide, population-based prevalence, comorbidity, 

treatment regimen, and direct medical cost of WD in South 

Korea from 2009 to 2013. Methods: Using 2 big data source 

from Health Insurance Review and Assessment Service (HIRA) 

claims database and Rare Intractable Diseases registration program 

database from 2009 to 2013 in Korea, we identified 

all patients with WD registered by physician as the International 

Classification of Diseases (ICD-10) code for WD (E83.0), 

and with the registration record (V174) for the rare disease in 

Korea. The five-year data included 1,509 patients linked with 

information including age, gender, comorbidity ICD-10 codes, 

prescribed medications, and direct medical costs. Results: The 

overall crude prevalence of WD was 2.38/100,000 people 

and it showed increasing tendency during the 5 years (2.27, 

2.28, 2.35, 2.45, and 2.53/100,000 at 2009-2013, respectively). 

Most (72%) patients were diagnosed before 30 years 

old, and 1.4% had additional disease code for neurologic/ 

psychologic disease. Liver cirrhosis and liver cancer were 

detected in 33.2% and 9.9%, respectively. Liver transplantation 

was performed in 4.6% of patients. D-penicillamine, trientine, 

and zinc were prescribed for 58.3%, 29.7%, and 13.9% 

of patients, respectively. Mean annual total medical cost per


person for WD was 1,643.15 USD. Conclusion: The prevalence 

of WD was slightly lower in Korean than other countries, 

showing an increase from 2009. Since about one third of WD 

patients had advanced liver disease such as cirrhosis and cancer, 

more efforts for early diagnosis and treatment for WD are 

warranted, especially during pediatric age. 

Disclosures: 

The following authors have nothing to disclose: Joo Yeong Baeg, Eun Sun Jang, 

Mo-ran Ki, Bo Hyun Kim, Kyung-ah Kim, Hwa Young Choi, Sook-Hyang Jeong 

2114 

Relapse of porphyria cutanea tarda after achieving 

remission: A Meta-analysis 

Maira Rizwan 1 , Karl Anderson 2 , Ashwani K. Singal 1 ; 1 Gastroenterology 

and Hepatology, University of Alabama, Birmingham, AL; 

2 university of texas, Galveston, TX 

Background: Porphyria cutanea tarda (PCT) is due to inhibition 

of hepatic uroporphyrinogen decarboxylase (UROD). 

Phlebotomy and 4-aminoquinolines (hydroxychloroquine and 

chloroquine, preferably low-dose) are effective treatment 

options. Data comparing relapse after documenting biochemical 

remission of PCT with these treatments are lacking. Methods: 

Pubmed and Embase databases were searched for studies 

reporting data on PCT relapse with minimum follow up for 

1 year after achieving remission with either phlebotomy or 

4-aminoquinolines. End point of treatment with phlebotomy 

was ferritin


gene, encoding alpha-1-antitrypsin (AAT) protein. AAT is a 

serine proteinase inhibitor that is synthesized in the liver and 

secreted into the bloodstream to protect the lungs from neutrophil 

elastase. Mutations in SERPINA1, most commonly the Z-allele 

(Z-AAT), cause the protein to misfold resulting in retention 

of aggregated AAT protein in hepatocytes and a corresponding 

decrease in circulating levels of AAT leading to Alpha-1-Antitrypsin 

Deficiency (A1ATD). Aggregation of Z-AAT protein in 

hepatocytes is associated with liver injury, fibrosis, cirrhosis, 

and hepatocellular carcinoma, while decreased circulating levels 

of AAT often presents as emphysema or chronic obstructive 

pulmonary disease. Using a lipid nanoparticle (LNP) system we 

developed for hepatic delivery of DsiRNAs, we demonstrate 

a significant reduction of human Z-AAT protein in the bloodstream 

and livers of the PiZ transgenic mouse model of hepatic 

A1ATD disease. Additionally, we provide evidence that targeting 

SERPINA1 with DsiRNA results in a reduction of A1ATD-related 

liver disease markers in PiZ transgenic mice. To further 

extend DsiRNA delivery approaches, we have also developed 

DsiRNA conjugates that can effectively deliver DsiRNAs to the 

liver without the need for LNP delivery systems. DsiRNA conjugates 

were prepared through the addition of N-acetyl galactosamine 

(GalNAc) sugars to the DsiRNA molecule to mediate 

delivery to hepatocytes via the highly expressed asialoglycoprotein 

receptor. SERPINA1-GalNAc conjugates delivered to 

PiZ transgenic mice effectively reduced Z-AAT protein levels. 

These results indicate Dicerna’s DsiRNA platform, delivered by 

either LNP-mediated or GalNAc-conjugated delivery systems, 

may be an effective therapeutic approach for hepatic disease. 

Disclosures: 

Rohan Diwanji - Employment: Dicerna Pharmceuticals; Stock Shareholder: 

Dicerna Pharmceuticals 

Utsav H. Saxena - Employment: Dicerna Pharmaceuticals 

Hank Dudek - Employment: Dicerna 

Nicole Avitahl-Curtis - Employment: Dicerna Pharmaceuticals, Inc. 

Chaitali Dutta - Employment: Dicerna Pharmaceuticals 

Benjamin Holmes - Employment: Dicerna Pharmaceuticals; Stock Shareholder: 

Dicerna Pharmaceuticals 

Marc Abrams - Employment: Dicerna Pharmaceuticals 

Kevin Craig - Employment: Dicerna Pharmaceuticals, Inc 

Luciano Apponi - Employment: Dicerna Pharmaceuticals 

Martin Koser - Employment: Dicerna Pharmaceuticals 

Chengjung Lai - Employment: Dicerna Pharmaceuticals 

Bob D. Brown - Employment: Dicerna Pharmaceuticals 

The following authors have nothing to disclose: Natalie W. Pursell, Wei Zhou, 

Weimin Wang, Dongyu Chen, Purva Pandya, Xue Shui, Boyoung Kim, Rachel 

Storr, Anee Shah, Edmond Chipumuro, Girish R. Chopda, Jr-Shiuan Yang, Marita 

S. Larsson Cohen, Naim Nazef, Nandini Venkat, Shanthi Ganesh, Venkat 

Krishnamurthy, Wendy Cyr, Zakir Siddiquee 

2117 

GNPAT Variant D519G in patients referred for HFE 

Hemochormatosis Testing 

Alexander Levstik 2 , Alan Stuart 2 , Paul Adams 1 ; 1 Gastroenterology, 

University Hospital, London, ON, Canada; 2 Laboratory Medicine, 

Western University, London, ON, Canada 

Background: Previous studies in high and low expressors has 

demonstrated that a variant in the GNPAT gene (D519G, 

Rs11558492, chromosome 1, exon 11) has been associated 

with severe iron overload in C282Y homozygotes for hemochromatosis 

(McLaren C, et al, Hepatology 2015). The GNPAT 

gene is associated with peroxisomal diseases and may be 

involved with transferrin receptor recycling. Gene silencing of 

GNPAT reduces intracellular hepcidin. In this study, a GNPAT 

variant was assessed prospectively in patients over a range of 

serum ferritin levels referred for HFE testing. Methods: Consecutive 

patients sent for HFE testing were studied for the GNPAT 

variant using a TaqMan kit assay (Life Technologies, Burlington, 

ON). The assay was validated by sequencing. Serum ferritin 

was compared in C282Y homozygotes with and without 

the GNPAT variant. The frequency of the GNPAT variant in 

referred patients was compared to a control population of voluntary 

blood donors without HFE mutations. Results: There were 

473 patients that had GNPAT analysis. The allele frequency for 

the GNPAT variant in C282Y homozygotes (n=81) was 0.302 

and in wild type control patients (n =392) was 0.213 (p = .08) 

Forty-nine percent (40) of the C282Y homozygotes were heterozygous 

(n=31) or homozygous (n = 9) for the GNPAT variant. 

The mean ferritin ± standard deviation did not significantly 

differ between C282Y homozygotes with (1450 μg/L ±1529) 

and without the GNPAT variant (1464 μg/L ± 1169). Conclusions: 

C282Y homozygotes referred for HFE testing commonly 

have a GNPAT variant. This GNPAT variant does not appear 

be a co-modifying gene affecting expression of HFE related 

hemochromatosis in this population. Further functional studies 

are in progress to determine the role of the GNPAT variant in 

cellular iron metabolism. 

GNPAT variant in patients 

Disclosures: 

The following authors have nothing to disclose: Alexander Levstik, Alan Stuart, 

Paul Adams 

2118 

Vitamin D deficiency promotes hepatocellular cancer 

through activation of WNT/TLR signaling with disruption 

of TGF-β/Smad3 

Ji-Hyun Shin 1 , Jian Chen 1 , Nina M. Muñoz 1 , Lior Katz 2 , Andrea 

C. Cortes 1 , Vivek Shukla 3 , Sangbae Kim 1 , H. Franklin Herlong 1 , 

Keigo Machida 4 , Hidekazu Tsukamoto 4 , Kirti Shetty 5 , Asif Rashid 1 , 

Wilma S Jogunoori 6 , Aiwu R. He 7 , Lynt B. Johnson 7 , Ju-Seog Lee 1 , 

Jon White 6 , Lopa Mishra 1 ; 1 The University of Texas MD Anderson 

Cancer Center, Houston, TX; 2 Sheba Medical Center, Tel 

Hashomer, Israel; 3 NCI, Bethesda, MD; 4 University of Southern 

California School of Medicine, Los Angeles, CA; 5 Johns Hopkins 

University School of Medicine, Baltimore, MD; 6 Veterans Affairs 

Medical Center, Washington, DC; 7 Georgetown University, Washington, 

DC 

Disruption of the TGF-β pathway has been associated with 

liver tumorigenesis, conditions associated with low Vitamin D 

(VD) levels. We examined a potential chemo-preventive role 

of VD in hepatocellular cancer (HCC) in the context of TGF-β 

inactivation. Methods: (1) We analyzed transcriptome profiles 

of 488 HCC and screened for somatic mutations (n = 202) and 

expression of the TGF-β pathway and VD pathway molecules 

(n = 147) in HCC from The Cancer Genome Atlas (TCGA). (2) 

Expression of TGF-β and VD pathway molecules was also evaluated 

by immunohistochemistry in liver samples from patients 

with hepatitis C virus (HCV)-associated cirrhosis who received 

VD supplements. (3) We examined the effects of VD on HCC 

development through gene and protein expression profiles in 

diethylnitrosamine -treated WT, and in mutant mouse models in 

the TGF-β pathway. (4) We further validated the effects of VD 

and performed further functional analyses examining crosstalk 

between TGF-β, VD and key members of cell proliferation and 

inflammation that were altered with deprivation of VD. Results: 

(1) We observed somatic mutations and a strong correlation 

between VD-related genes and the TGF-β pathway in the TCGA 

genomic analysis. (2) We observed low levels of TGF-β path-


way member expression profiles with activation of β-catenin 

in tissues from HCV cirrhotic patients not on VD supplementation, 

while those supplemented showed restoration of TGF-β 

pathway member expression profiles with lower expression 

of β-catenin. (3) Strikingly, we observed a threefold higher 

incidence of HCC in Smad3 mice deprived of VD compared 

to mutant and WT mice fed a high-VD diet. (4) RPPA analyses 

revealed an activation of β-catenin, Stat5A, and Bcl2-XL, with 

reduced levels of the tumor suppressor PDCD4 in Smad3 +/- 

mice deprived of VD diet. (5) VD treatment suppressed HCC 

cell proliferation and significantly reduced expression levels 

of TLR7 and TLR-mediated inflammation associated genes in 

Smad3 -/- MEFs. (6) Functional analyses revealed that loss of 

Smad3 and shRNA for VD receptor, induced nuclear localization 

and activation of β-catenin. (5) Luciferase and ChIP assays 

showed that TGF-β negatively regulates TLR7 transcriptional 

activity through Smad3 binding at Smad Binding Elements at 

the TLR7 promoter. Conclusions: We conclude that VD suppresses 

proliferation of hepatocellular cancer cells by restoring 

TGF-β signaling. VD deficiency promotes tumor growth in the 

context of Smad3 disruption potentially through regulation of 

TLR7 expression and β-catenin nuclear localization. VD could 

therefore be a strong candidate for hepatocellular cancer prevention 

in the context of aberrant Smad3 signaling. 

Disclosures: 

Hidekazu Tsukamoto - Consulting: Suntory Ltd.; Grant/Research Support: The 

Toray Co. 

The following authors have nothing to disclose: Ji-Hyun Shin, Jian Chen, Nina M. 

Muñoz, Lior Katz, Andrea C. Cortes, Vivek Shukla, Sangbae Kim, H. Franklin 

Herlong, Keigo Machida, Kirti Shetty, Asif Rashid, Wilma S Jogunoori, Aiwu R. 

He, Lynt B. Johnson, Ju-Seog Lee, Jon White, Lopa Mishra 

2119 

Chemokine CCL3/CCR5-recruited, Cytotoxic CD4 + T 

Cells Eradicate Hepatoma Cells in a Model of Anticancer 

Chemotherapy with High-Dose Cyclophosphamide 

Tatsushi Naito 1,2 , Tomohisa Baba 2 , Kazuyoshi Takeda 3 , Soichiro 

Sasaki 2 , Naofumi Mukaida 2 , Yasunari Nakamoto 1 ; 1 Second 

Department of Internal Medicine, University of Fukui, Fukui, Japan; 

2 Division of Molecular Bioregulation, Cancer Research Institute, 

Kanazawa University, Kanazawa, Japan; 3 Department of immunology, 

Juntendo University, School of Medicine, Tokyo, Japan 

BACKGROUND: The mortality rate of hepatocellular carcinoma 

(HCC) is high because of frequent recurrence due to 

the high carcinogenic potentials of background chronic liver 

inflammation and cirrhosis. Thus, novel therapeutic strategy 

is required including immune therapy. Today, accumulating 

evidences indicate the potential role of CD4 + T cells with direct 

cyototoxicity (CD4 + CTLs) in antitumor immunity. Correlation 

between decrease of CD4 + CTLs and high mortality is also 

reported in patients with HCC. However, the importance of 

CD4 + CTLs in patients treated with anticancer chemotherapy 

remains to be elucidated. Hence, we examined the role of 

CD4 + CTLs, in chemotherapy-induced eradication of hepatoma 

by means of an animal HCC model. METHODS and RESULTS: 

We inoculated a murine hepatoma cell line, BNL 1ME A.7R.1 

(BNL), subcutaneously into Balb/c mice and gave single intraperitoneal 

injection of 150 mg/kg cyclophosphamide (CTX) 

after tumor formation. CTX treatment eradicated tumors in wildtype 

(WT) mice, whereas none of the tumors disappeared in 

T cell lacking nude mice. Next, we depleted CD4 + or CD8 + 

cells by antibody to clarify which subset was involved in the 

efficacy of CTX. We found CD4 + but not CD8 + cell depletion 

abrogated CTX-induced tumor eradiation. Consistently, CTX 

treatment increased intratumoral CD4 + CTLs, which expressed 

cytolytic granule molecule, CD107a and granzyme B observed 

by flowcytometry and immunofluorescence analysis. To delineate 

the underlying mechanism of increase in CD4 + CTLs, congenic 

CD45.1 splenocytes were transferred to tumor-bearing 

CD45.2 mice one day after CTX treatment and tumors were 

analyzed with time by flowcytometry. Transferred CD4 + cells 

were recruited into tumors on day 3, and the recruited cells 

expressed CD107a without antigen presentation at draining 

lymph nodes and proliferation in tumor tissues. Moreover, CTX 

administration enhanced mRNA expression of CC chemokine 

CCL3 in tumor tissues, and CTX-mediated tumor regression 

was attenuated in mice deficient in CCR5 gene, the receptor 

for this chemokine. Consistently, CTX-induced accumulation of 

intratumoral CD107a-expressing CD4 + T cells was less in mice 

receiving CCR5-deficient mouse-derived splenocytes than those 

receiving WT mouse-derived splenocytes. CONCLUSIONS: CC 

chemokine CCL3/CCR5 plays an important role in intratumoral 

CD4 + CTL migration in the process of CTX-induced tumor eradication. 

The results suggest that the CC chemokine-dependent 

mechanism may be a plausible strategy to enhance the antitumor 

cytotoxicity in the treatment of HCC with chemotherapy. 

Disclosures: 

The following authors have nothing to disclose: Tatsushi Naito, Tomohisa Baba, 

Kazuyoshi Takeda, Soichiro Sasaki, Naofumi Mukaida, Yasunari Nakamoto 

2120 

IFN-α stimulates IFN-γ expression in type I NKT cells 

and enhances the inhibition of HCV replication in 

human hepatocyte chimeric mice 

Eisuke Miyaki 1 , Michio Imamura 1 , Nobuhiko Hiraga 1 , Takuro 

Uchida 1 , Hiromi Kan 1 , Masataka Tsuge 1 , Hiromi Abe 1 , C. Nelson 

Hayes 1 , Hiroshi Aikata 1 , Chise Tateno 2 , Kazuaki Chayama 1 ; 

1 Department of Gastroenterology and Metabolism, Applied Life 

Sciences, Institute of Biomedical & Health Sciences, Hiroshima 

University, Hiroshima, Japan; 2 PhoenixBio Co., Ltd., Higashihiroshima, 

Japan 

Background & Aims: Interferon (IFN) inhibits hepatitis C virus 

(HCV) replication through up-regulation of IFN-stimulated gene 

(ISG) expression. However, the effect of IFN on immune cells is 

not well known. In this study, we analyzed the immune cell-mediated 

antiviral effects of IFN-α using HCV-infected mice. Methods: 

Genotype 1b HCV-infected urokinase-type plasminogen 

activator (uPA)-severe combined immunodeficiency (SCID) mice 

with transplanted human hepatocytes were injected intra-peritoneally 

with 2 x 10 7 human peripheral blood mononuclear 

cells (PBMCs) obtained from a healthy volunteer. The mice then 

received daily intra-peritoneal injections with 1000 IU/g of 

IFN-α for seven days, during which mouse serum HCV RNA, 

human albumin and cytokine levels were analyzed. Flow cytometric 

analysis of liver infiltrating human immune cells and 

histopathological examination were performed. Results: Seven 

days of IFN-α treatment without human PBMC injection in mice 

reduced serum HCV RNA levels by 1.23 ± 0.15 log whereas 

in combination with PBMCs, HCV RNA levels were reduced 

by 2.5 ± 0.45 log (p


these mice, the anti-HCV effect and development of human 

mononuclear cell chimerism by human PBMCs and IFN-α treatment 

was completely negated, suggesting that IFN-γ produced 

by NKT cells is essential not only for its anti-viral effect, but also 

for the development of human lymphocyte chimerism in human 

PBMCs and IFN-α treated mice. Conclusion: IFN-α stimulates 

IFN-γ expression in type I NKT cells and enhances the inhibition 

of HCV replication. Type I NKT cells might represent a new 

therapeutic target for chronic hepatitis C patients. 

Disclosures: 








The following authors have nothing to disclose: Eisuke Miyaki, Michio Imamura, 

Nobuhiko Hiraga, Takuro Uchida, Hiromi Kan, Masataka Tsuge, Hiromi Abe, C. 

Nelson Hayes, Hiroshi Aikata, Chise Tateno 

2121 

Small specific-sized Hyaluronic Acid normalizes 

TLR4-mediated signaling in Kupffer cells after chronic 

ethanol exposure associated with M2 polarization 

Paramananda Saikia 1 , Katherine A. Pollard 1 , Megan R. McMullen 

1 , Laura E. Nagy 1,2 ; 1 Center For Liver Disease Research, Pathobiology, 

Lerner Research Institute, Cleveland Clinic, Cleveland, 

OH; 2 Molecular Medicine, Case Western Reserve University, 

Cleveland, OH 

Hyaluronan (HA) is the major glycosaminoglycan in the extracellular 

matrix and ubiquitously present in many tissues. While 

HA has been used as a biomarker for liver injury for decades, 

the contribution of HA to alcoholic liver disease is not well 

understood. During acute and chronic inflammation or tissue 

injury, reactive oxygen species and matrix metalloproteinases 

increase HA turnover, resulting in local and systemic accumulation 

of HA fragments of different molecular weights. HA 

is recognized as an important damage associated pattern 

molecule (DAMP) that regulates innate immunity. However, 

depending on their size, HA fragments can have either pro-inflammatory 

or anti-inflammatory functions. Chronic alcohol 

consumption is associated with an increase in the sensitivity of 

hepatic macrophages to signaling via TLR4 associated with an 

M1 polarization and enhanced expression of pro-inflammatory 

cytokines. Here we investigated the impact of specific-sized 

small HA on TLR4 mediated signaling in Kupffer cells after 

chronic ethanol exposure to rats. Primary cultures of rat Kupffer 

cells were isolated from rats chronically exposed to ethanol 

via the Lieber-DeCarli diet or pair-fed control diets. Kupffer 

cells from ethanol-fed rats were more sensitive to stimulation 

with LPS, resulting in increased expression of mRNA for the 

pro-inflammatory cytokine, TNFα. When Kupffer cells were 

pre-treated with HA fragments for 5 hr prior to LPS challenge 

for 60 min, specific small-sized HA normalized TNFα mRNA 

expression in Kupffer cells from ethanol fed rats. In contrast, 

higher molecular weight HA fragments had no effect on TNF-α 

production of Kupffer cells. Normalization of TLR4 signaling 

after treatment with small-sized HA was associated with an 

increase expression of the M2 phenotypic gene arginase-1 

(arg-1) and a decrease expression of the M1 associated gene 

iNOS. In summary, these data demonstrate for the first time 

that specific small-sized HA fragments promote polarization 

of Kupffer cells to M2/anti-inflammatory phenotype to normalize 

chronic ethanol induced sensitization of TLR4 signaling in 

Kupffer cells. 

Disclosures: 

The following authors have nothing to disclose: Paramananda Saikia, Katherine 

A. Pollard, Megan R. McMullen, Laura E. Nagy 

2122 

Novel in vivo and in vitro models of Hepatitis E virus 

genotype 3 infectivity for chronic human HEV infection. 

Martijn D. van de Garde 1 , Suzan D. Pas 3 , Guido van der Net 3 , 

Bart L. Haagmans 3 , Andre Boonstra 1 , Thomas Vanwolleghem 1,2 ; 

1 Department of Gastroenterology and Hepatology, Erasmus MC 

University Hospital, Rotterdam, Netherlands; 2 Gastroenterology 

and Hepatology, University Hospital Antwerp, Antwerp, Belgium; 

3 Viroscience, Erasmus Medical Center, Rotterdam, Netherlands 

Hepatitis E virus (HEV) genotype 3 (gt3) infections are emerging 

in western countries. The pathogenesis of HEV infection 

with associated liver pathology and clinical disease is poorly 

understood due to a lack of suitable model systems. Here we 

apply a novel in vivo and in vitro model system to characterize 

the infectivity of different HEV RNA containing human clinical 

samples. Human-liver chimeric mice (uPA +/+ Nod-SCID-IL2Rγ -/- ) 

and human lung adenocarcinoma A549 cells were challenged 

with HEV gt3 obtained from human plasma, feces or liver-biopsy 

of 3 patients. Detection and quantification of HEV RNA 

was performed in mouse serum, feces, bile and liver or in 

A549 culture supernatant using RT-qPCR. High HEV RNA levels 

(up to 8 log IU HEV RNA/gr) were consistently detected in 

100% of chimeric mouse livers (n=10) from week 2-14 post 

inoculation with human feces-derived HEV (6-8 logs IU). Feces 

of these mice was positive for HEV RNA at least once during 

follow-up, while HEV viremia was inconsistently detectable, 

with maximum viral loads of 3.6 log IU/ml. HEV derived from 

a cryopreserved liver biopsy (5.5 log IU) similarly resulted in 

moderate to high HEV RNA levels in mouse feces, bile and 

liver (n=2). In contrast, anti-HEV IgG/IgM negative, HEV RNA 

positive plasma (6 log IU) was not infectious in any of the 

inoculated chimeric animals (n=8). In these animals human 

albumin plasma levels were stable and comparable to HEV-infected 

chimeric mice, indicating a functional hepatocyte graft. 

Infection of A549 cells with HEV RNA positive clinical samples 

showed rapid and increasing HEV RNA titers in culture 

supernatant within 7 days and could be maintained for more 

than 7 passages. Consistent in vitro HEV infectivity was seen 

with feces samples of all 3 patients, while only 2 of 3 human 

serum samples could be cultured in vitro, showing less efficient 

propagation. In conclusion, infectivity of feces derived human 

HEV is higher compared to serum derived HEV both in vitro 

and in vivo. The sustained HEV gt 3 liver infections induced in 

chimeric mice, show preferential viral shedding towards mouse 

bile and feces, mimicking the course of infection in humans. 

Besides antiviral efficacy studies, both models will guide future 

investigations into the biology of HEV infectivity with implications 

towards transfusion transmitted HEV in humans. 

Disclosures: 


Roche, Gilead 

The following authors have nothing to disclose: Martijn D. van de Garde, Suzan 

D. Pas, Guido van der Net, Bart L. Haagmans, Thomas Vanwolleghem


2123 

Liver Plasmacytoid Dendritic Cells Expressing miR-181 

prolong graft survival by increasing T Regulatory cells 

and Decreasing B cells as Revealed by Mass Cytometry 

(CyTOF 

Audrey H. Lau 1 , Matthew J. Vitalone 2 , Xiumei Qu 2 , Todd Shawler 2 , 

Olivia M. Martinez 2 , Carlos O. Esquivel 2 , Sheri M. Krams 2 ; 1 Pediatrics, 

Division of Gastroenterology, Hepatology, and Nutrition, 

Stanford/LPCH, San Francisco, CA; 2 Surgery, Stanford University, 

Stanford, CA 

Liver allografts are well tolerated and other solid organ 

allografts, when transplanted concurrently with livers, show 

improved outcomes. However, the mechanisms underlying 

“hepatic tolerance” have yet to be elucidated. Previous data 

show that hepatic dendritic cells (DC) have diminished antigen 

presentation and immune stimulatory function compared 

with DC in lymphoid tissue. Immature plasmacytoid (p)DC have 

been shown to induce graft prolongation and we have previously 

shown miR-181a1b1 is increased in pDC as compared 

to conventional DC. Wild-type (WT) BALB/c mice were transplanted 

with allogeneic (C57BL/6) vascularized heterotopic 

heart grafts. Animals received either no treatment (n=3) or were 

injected i.v. with 0.5×10 6 hepatic pDC (n=5) or miR-181a1b1 -/- 

(KO) pDC (n=3) at day (D)−7 from transplant. Recipients with 

no treatment rejected their allograft by D7 while recipients pretreated 

with hepatic pDC had significantly prolonged allograft 

survival to D15-21 (p1.5 

yrs) Opn -/- mice, which displayed significant hepatomegaly 

and splenomegaly compared to their age-matched WT littermates. 

Morphological features consisted of small aggregates 

of cells with intensely basophilic nuclei and clusters of immature 

and mature myeloid cells located in the sinusoids and in 

the portal areas. Accumulation of HSCs occurred also in the 

spleen but to a much lesser extent in the kidney and in the lung. 

Furthermore, Opn -/- mice showed premature exit of immature 

hematopoietic cells from the bone marrow to the spleen and 

to the liver, increased cellularity of lineage-negative cells and 

elevated hepatic GCSF, CXCL1, MCSF, MIP1α, RANTES and 

TNFα as well as serum MCSF and CXCL1 compared to WT 

littermates. Opn -/- mice also presented numerous iron deposits 

in the periportal zone of the liver, in the bone marrow and in 

the spleen but they were minimal in the lung and were absent 

in the heart and in the kidney, suggesting dysregulation of iron 

metabolism in the absence of Opn compared to WT littermates. 

CONCLUSION: These results suggest a critical role for OPN in 

HSC homeostasis by regulation of bone marrow homing and 

modulation of cytokine levels involved in their development. 

Disclosures: 

The following authors have nothing to disclose: Fernando Magdaleno, Xiadong 

Ge, Holger Fey, Costica Aloman, M. Isabel Fiel, Natalia Nieto 

2125 

IL-17A plays a pivotal role in LPS/GalN-induced fulminant 

hepatic injury in mice. 

Hiroshi Kono, Shinji Furuya, Chao Sun, Hideki Fujii; First Department 

of Surgry, University of Yamanashi, Chuo, Japan 

Background Lipopolysaccharide/D-Galactosamine (LPS/Gal- 

N)-induced hepatic injury is an experimental model of fulminant 

hepatic failure in which tumor necrosis factor-a (TNF-a) plays 

a pivotal role. Moreover, it was reported from our laboratory 

that interleukin (IL)-17A enhanced production of TNF-a by the 

Kupffer cell. Objective The purpose of this study was to determine 

the role of IL-17A in LPS/GalN-induced hepatic injury 

in mice. Methods LPS/GalN was injected into three mouse 

models: wild-type (WT) mice, IL-17A knockout (KO) mice, or 

IL-17A knockout mice treated with recombinant mouse (rm) 

IL-17A homodimer (KO + rmIL-17A). Survival was assessed 

for 24 hours after LPS/GalN injection, and histopathologic 

findings were evaluated at various time points after LPS/GalN 

injection for neutrophil and apoptosis markers. After LPS/GalN 

injection, expression of the inflammatory mediators TNF-a, IL-6, 

monocyte chemotactic protein (MCP)-1, IL-17A, high mobil-


ity group box 1 (HMGB1) and soluble intercellular adhesion 

molecule (sICAM)-1 was assessed in serum by enzyme-linked 

immunosorbent assay (ELISA). Results In the WT mice, mortality 

was 50 % at 24 hours after LPS/GalN injection. In contrast, 

all KO mice survived at 24 hours after injection. There were 

significant differences in the mortality between the WT mice 

and the KO mice. However, in KO + rmIL-17A mice, mortality 

was not significantly different compared to the other groups. 

Neutrophil infiltration and apoptosis were significantly greater 

in WT mice than KO mice. Furthermore, liver injury was severe 

in the WT mice compared with the KO mice; however, in KO 

+ rmIL-17A mice, injury was similar to those in the WT mice. 

Serum ALT levels were also significantly greater in WT mice 

than KO mice. In KO + rmIL-17A mice, these levels were similar 

to those in WT mice, as expected. Supporting the pathological 

findings, serum TNF-a, MCP-1, IL-17A, HMGB1 and sICAM-1 

levels were also significantly greater in WT mice than KO mice. 

In KO + rmIL-17A mice, these levels were similar to those in 

WT mice. Conclusions IL-17A is a key regulator in hepatic 

injury caused by neutrophil-induced inflammatory responses 

after LPS/GalN injection. The lack of IL-17A decreases the 

serum cytokine and chemokine levels, and all animals survived 

after LPS/GalN treatment. These results suggest that IL-17A 

may play one of the important role in fulminant hepatic injury. 

Disclosures: 

The following authors have nothing to disclose: Hiroshi Kono, Shinji Furuya, 

Chao Sun, Hideki Fujii 

2126 

Upregulated hepatic expression of pattern recognition 

receptors and Th1 type cytokine genes are associated 

with clearance of HEV in genotype 1 infected rhesus 

macaques 

Youkyung Choi, Xiugen Zhang, Brianna Skinner, Chong-Gee Teo; 

Center for Disease Control and Prevention, Atlanta, GA 

Infection by hepatitis E virus (HEV) genotype 1 causes acute 

liver disease that is usually self-limited but occasionally leads 

to fulminant hepatic failure. Immune responses, rather than 

virus-induced hepatocytolysis, have been shown to relate to 

the pathogenesis of acute hepatitis E and liver failure. Since 

HEV-specific CTL responses do not appear to be robustly 

induced in peripheral blood of HEV-infected patients during the 

acute and convalescent phases of infection, the liver is likely to 

be the major site of pathogenically relevant anti-HEV immune 

responses. In order to investigate the host immune responses 

induced by HEV infection, host gene expression profiles were 

analyzed in liver biopsy tissues of experimentally infected rhesus 

macaques. Three rhesus macaques were inoculated with 

HEV genotype 1 and observed for up to 150 days. All 3 exhibited 

the course typical of acute hepatitis. Thus, HEV RNA was 

detected in stools from 8 to 13 days after infection (DAI) and 

in serum from DAIs 22 to 32, becoming undetectable between 

DAIs 51 and 140; and elevated ALT activity and IgM and IgG 

anti-HEV antibody became detectable between DAIs 39 and 

56. Hepatic gene expression profiles were studied using 3 different 

real-time PCR based rhesus-macaque-specific arrays (Qiagen) 

that included 250 immune response-related genes. PCR 

arrays were done at 3 time-points: during and after the peak 

of stool HEV RNA detectability, and when HEV RNA became 

undetectable. Levels of gene expression were compared to 

normal liver tissues obtained from a macaque seronegative for 

anti-HEV. Genes showing ≥2-fold change in expression were 

considered to be significantly altered by HEV infection. Up to 

57 genes were up-regulated during and after the peak of stool 

HEV RNA detectability. These 57 genes could be grouped to 

belong to 3 broad categories: pattern recognition receptors 

(PRRs) (including TLR7 and TLR9); those associated with type 1 

interferon response (including ISG15, ISG20, IRF1, IRF5 and 

IRF7); and genes encoding Th1 type cytokines (including IL-12, 

CCR5, CXCR3, IFNg, STAT4 and TBX21). Conclusions: Genes 

upregulated in the liver during HEV infection include those 

associated not only with type 1 interferon response and Th1 

type cytokines but also PRRs. Upregulated hepatic PRR genes 

possibly mediate heightened interferon response and cytokine 

secretion, thereby contributing to resolution of acute infection. 

Studies are ongoing to study the kinetics, in plasma, of proteins 

associated with these responses. 

Disclosures: 

The following authors have nothing to disclose: Youkyung Choi, Xiugen Zhang, 

Brianna Skinner, Chong-Gee Teo 

2127 

Hepatocellular carcinoma is accelerated by modified 

western diet involving M2 macrophage polarization 

mediated by HIF-1a activation 

Aditya Ambade, Abhishek Satishchandran, Banishree Saha, 

Karen Kodys, Gyongyi Szabo; Medicine, UMass Medical School, 

Worcester, MA 

Background/Aims: Obesity is an independent risk factor for 

the development of hepatocellular carcinoma (HCC), the third 

leading cause of cancer related deaths worldwide. Tissue resident 

macrophages play a key role in promoting tumor progression. 

Transformed cells within the tumor tissue can induce 

macrophage polarization as a survival mechanism. However, 

the effect of western diet on macrophage polarization in tumor 

tissue microenvironment is not known. Here we sought to understand 

the role of western diet in promoting tumor development 

and inducing macrophage polarization after carcinogen 

pre-exposure. Methods: Four week-old male C57BL/6 mice 

were injected with 6 doses of N, N diethyl nitrosamine (DEN) 

intraperitoneally. 75 mg/kg/week DEN was administered for 

3 weeks followed by 100 mg/kg/week for 3 weeks. At 6 

weeks of age, mice were divided into modified western diet 

containing high fat-high cholesterol-high sugar (HF-HC-HS) 

and chow (control) diet groups. All mice were sacrificed at 

30 weeks of age; blood, liver, primary hepatocytes and liver 

mononuclear cells (LMNCs) were collected. In vitro, naïve primary 

hepatocytes were treated with 10 mM DEN and/or 330 

mM palmitic acid (PA) and supernatants were transferred to 

RAW 264.7 macrophages. Results: HF-HC-HS diet resulted 

in weight gain except for mice with DEN injection. ALT and 

liver to body weight ratio were significantly higher in HF-HC- 

HS+DEN compared to HF-HC-HS diet mice. Serum AFP levels 

were significantly higher in HF-HC-HS+DEN mice compared to 

all other groups (P


hepatocyte supernatants induced HIF-1a DNA binding and 

up-regulated VEGF in RAW 264.7 macrophages. Conclusion: 

Modified western diet accelerated carcinogen initiated hepatocellular 

carcinoma, involving hepatocytes derived signals promoting 

M2 macrophage polarization. 

Disclosures: 

The following authors have nothing to disclose: Aditya Ambade, Abhishek Satishchandran, 

Banishree Saha, Karen Kodys, Gyongyi Szabo 

2128 

CD8 + T-cells drive liver injury and hepatocellular carcinoma 

in chronic liver disease 

Jessica Endig 1 , Laura E. Buitrago 1 , Silke Marhenke 1 , Anna Saborowski 

1 , jutta Schütt 2 , Florian Reisinger 3 , Florian Limbourg 1 , 

Christian Koenecke 1 , Alina Schreder 1 , Robert Geffers 4 , Michael 

P. Manns 1 , Mathias Heikenwälder 5 , Thomas Longerich 6 , Arndt 

Vogel 1 ; 1 Hannover Medical School, Hannover, Germany; 

2 Philipps University Marburg, Marburg, Germany; 3 Technische 

Universität München, Munich, Germany; 4 Helmholtz Centre for 

Infection Research, Braunschweig, Germany; 5 Helmholtz Zentrum 

München, Munich, Germany; 6 Aachen University Hospital, 

Aachen, Germany 

Tumors frequently arise at sites of inflammation and chronic 

inflammation is increasingly recognized as a key factor in 

the pathogenesis of many malignancies. Hepatocellular carcinoma 

(HCC), one of the most lethal and prevalent cancers 

worldwide, represents a classic example of inflammation-linked 

cancer. Although activation of different cytokines has been 

reported in chronic liver disease, the critical components linking 

inflammation and hepatocarcinogenesis remain elusive. To 

characterize the role of the immune system in hepatic injury 

and tumor development, we comparatively studied the extent 

of liver disease and hepatocarcinogenesis in immunocompromised 

vs. immunocompetent Fah-deficient mice. Flares of liver 

injury were repeatedly induced which led to HCC formation 

within 3-4 months. Strikingly, chronic liver injury and tumor 

development were markedly suppressed in alymphoid Fah -/- 

mice despite an overall increased mortality. Mechanistically, 

we show that CD8 + Tcell-derived lymphotoxin-β (LTβ) is a 

central mediator of HCC formation in Fah -/- mice. Pharmacological 

inhibition of the lymphotoxin-β receptor (LTβR) indeed 

delays tumors development in mice with chronic liver injury 

without preventing liver damage. We here demonstrate that 

lymphocytes play a decisive, yet ambiguous role in chronic 

liver disease: while infiltrating lymphocytes mediate hepatocyte 

damage, liver fibrosis and tumor development, they also protect 

mice from acute on chronic liver failure and are required 

for activation of liver progenitor cells during regeneration. Our 

data illustrate that the immune system needs to be tightly regulated 

in a context-specific fashion, in order to balance immune 

surveillance and cancer risk. We propose that targeting specifically 

the tumor-promoting pathways such as LTβ might be an 

attractive chemopreventive strategy for patients at risk. 

Disclosures: 





Novartis 

The following authors have nothing to disclose: Jessica Endig, Laura E. Buitrago, 

Silke Marhenke, Anna Saborowski, jutta Schütt, Florian Reisinger, Florian 

Limbourg, Christian Koenecke, Alina Schreder, Robert Geffers, Mathias Heikenwälder, 

Thomas Longerich, Arndt Vogel 

2129 

Combined bone marrow plus liver transplantation in 

cynomolgus monkeys 

Joshua Weiner, Yojiro Kato, Sulemon Chaudhry, Raimon Duran- 

Struuck, Mercedes Martinez, Paula Alonso-Guallart, Jonah Zitsman, 

Jay H. Lefkowitch, Tomoaki Kato, Megan Sykes, Adam D. 

Griesemer; Columbia Center for Translational Immunology, New 

York Presbyterian-Columbia University Medical Center, New York, 

NY 

Introduction Combined kidney/bone marrow transplantation 

(CKBMT) induces tolerance in both primates and humans. 

Since liver is thought to be more tolerogenic than kidney, we 

hypothesized that combined liver/bone marrow transplantation 

(CLBMT) should be successful. We performed this pre-clinical 

study in cynomolgus monkeys. Methods Five pairs of animals 

were used. All were mismatched for class I, and the first 2 pairs 

were mismatched for class II. Liver and ~3 x 10^8 donor bone 

marrow cells/kg were transplanted on day 0. The induction 

regimen consisted of 1.5Gy total body irradiation on day-6, 

ATGAM (Pfizer) (50mg/kg) on days -2, -1, and 0, rituximab 

(Genentech) (20mg/kg) on day -6 (Recipients 2-5, also day 

6 for Recipients 4 and 5), 7Gy thymic irradiation on day -1, 

splenectomy on day 0, and 28 days of either IM cyclosporine 

(Novartis) or continuous IV tacrolimus (Astellas) followed by 

4-week taper (tacrolimus only). Recipients 4 and 5 additionally 

received LoCD2 (Mass Biologics) and anti-CD40 mAb (Mass 

Biologics). Anti-donor MLR was assessed by plating recipient 

and irradiated donor cells (1 x 10^6 each) for 5 days and 

pulsing with tritiated thymidine for 24 hours. Results Recipients 

1-3 survived 42, 69, and 57 days with multilineage mixed 

chimerism (MC) up to days 36, 40, and 23 respectively. They 

had normal to increased anti-donor cellular responses, except 

Recipient 1, and severe histological cellular rejection. Flow 

cytometry showed high percentages of CD8 T effector memory 

cells (TEM) in the blood, graft, and draining node but not 

peripheral nodes. Therefore, we added costimulatory blockade 

(anti-CD40) and depletion of CD2+ memory cells (LoCD2b) 

for Recipients 4 and 5, and subsequently this cell population 

did not expand. In contrast to the first three recipients, the 

fourth recipient survived 61 days (final 3 weeks with negligible 

immunosuppression levels) and died of diarrhea/electrolyte 

abnormalities on day 61 with normal LFTs, no histological 

rejection or GVHD, significant MC, and decreased anti-donor 

cellular responses. Recipient 5 is now 40 days post-transplantation 

with 70-80% chimerism in all lineages and no signs 

of rejection despite weaning immunosuppression. Conclusion 

CLBMT induces transient MC, but rejection eventually occurs. 

High levels of CD8 T effector memory cells likely contribute. 

However, when additional costimulatory blockade and CD2 

depletion are given, these cells are significantly depleted, preventing 

rejection, prolonging MC, decreasing anti-donor cellular 

responses, and possibly permitting tolerance. 

Disclosures: 

Tomoaki Kato - Grant/Research Support: Novartis 

The following authors have nothing to disclose: Joshua Weiner, Yojiro Kato, 

Sulemon Chaudhry, Raimon Duran-Struuck, Mercedes Martinez, Paula Alonso-Guallart, 

Jonah Zitsman, Jay H. Lefkowitch, Megan Sykes, Adam D. Griesemer


2130 

Allogeneic and xenogeneic transplantation of primary 

hepatocytes into the fetal liver in mice 

Yoshinobu Saito, Hayato Hikita, Yasutoshi Nozaki, Yugo Kai, Yuki 

Makino, Tasuku Nakabori, Satoshi Tanaka, Satoshi Aono, Ryotaro 

Sakamori, Naoki Hiramatsu, Tomohide Tatsumi, Tetsuo Takehara; 

Gastroenterology and Hepatology, Osaka University Graduate 


Background and Aim: Liver humanized mouse models using 

Alb-uPA SCID mice and Alb-HSVtk NOG mice have been 

reported to provide a lot of information on human liver physiology 

and pathology. However, these mouse models are not 

appropriate for investigating chronic inflammation and liver 

cancer because these mice are immune-deficient and could not 

survive more than one year. Immune tolerance is considered 

to be established during embryonic or neonatal stage. In this 

study, we investigated the engraftment efficacy of allogeneic or 

xenogeneic hepatocytes into the fetal liver by in utero transplantation 

in immune-competent mice. Method: As immune-competent 

recipient mice, we used hepatocyte-specific Mcl-1 knockout 

(KO) mice, which cause spontaneous hepatocyte apoptosis 

in life (Liver Injury mice), and hepatocyte-specific Mcl-1 and 

Bcl-xL double KO (DKO) mice, which show a decreased number 

of hepatocytes on embryonic (E) 18.5 day and die within 

one day after birth due to hepatic failure (Liver Impairment 

mice). Primary hepatocytes isolated from ubiquitously green fluorescent 

protein (GFP) transgenic (Tg) mice or primary human 

hepatocytes were administered into these recipient mice on E 

16.5 day via vitelline vein. These embryos were given birth by 

cesarean section and these livers were investigated at birth or 

at the age of 6 weeks. Results: Hepatocyte-transplanted wildtype 

mice and Liver Injury mice showed scattered embolization 

areas at birth by HE staining of the liver sections. The embolization 

was considered to be caused by transplanted hepatocytes. 

In contrast, Liver Impairment mouse livers escaped from 

embolization. In livers of wild-type mice and Liver Injury mice 

transplanted with GFP Tg hepatocytes, GFP-positive cells were 

observed forming a lot of clusters. While repopulation rate was 

not different between livers of wild-type mice and those of Liver 

Injury mice at birth (35.8% (3.6-48.3) and 29.8% (20.4-39.1), 

respectively), that of Liver Impairment was 7.8% (3.3-16.7). 

Meanwhile, repopulation rate at the age of 6 weeks was 0% in 

wild-type mice and 4.2% (2.1-8.0) in Liver Injury mice. In livers 

of wild-type and Liver Injury mice transplanted with primary 

human hepatocytes, embolization was similarly observed at 

birth by HE staining, suggesting that transplanted hepatocytes 

might be engrafted. Conclusion: Our results suggests that transplantation 

of primary hepatocytes into the fetal liver of Liver 

Injury mice, but not Liver Impairment mice, have the possibility 

of generating humanized liver mice without immunodeficiency. 

Disclosures: 






The following authors have nothing to disclose: Yoshinobu Saito, Yasutoshi 

Nozaki, Yugo Kai, Yuki Makino, Tasuku Nakabori, Satoshi Tanaka, Satoshi 

Aono, Ryotaro Sakamori, Naoki Hiramatsu, Tomohide Tatsumi 

2131 

The gut-liver axis coordinate T-cell-mediated immunity 

by regulating the antigen-presenting activity of macrophages 

in murine liver 

Nobuhito Taniki 1 , Nobuhiro Nakamoto 1 , Hirotoshi Ebinuma 1 , 

Po-sung Chu 1 , Takeru Amiya 1 , Shunsuke Shiba 1 , Akihiro Yamaguchi 

1 , Yuko Wakayama 1 , Hidetsugu Saito 2 , Takanori Kanai 1 ; 

1 Internal Medicine, Keio University, School Of Medicine, Tokyo, 

Japan; 2 Pharmaceutical Department, Keio University, Tokyo, Japan 

BACKGROUND & AIMS: The liver is a barrier for gut-derived 

antigens from degraded commensal bacteria. In colitic mice, 

live bacteria escaping the gut enter the liver through the portal 

vein. Specific subsets of innate immune cells suppress immune 

responses to such gut-derived antigens. The hygiene hypothesis 

proposes the recent increase in autoimmune hepatitis is the 

result of modern highly hygienic lifestyles. Reduced antigen 

exposure may cause dysfunctional hepatic immunosuppressive 

mechanisms. We hypothesized that increased antigen stimuli 

might activate immunosuppression. We investigated the role of 

innate immune cells in immunological tolerance in murine liver 

and the underlying mechanisms, focusing on the gut-liver axis. 

METHODS: We reported that dextran sulfate sodium (DSS)- 

treated mice (a colitis model) developed mild liver inflammation 

by histology and serology compared with normal mice following 

a sub-lethal dose of concanavalin A (ConA) (a T-cell-mediated 

hepatitis model). Colitis severity was inversely correlated 

with subsequent liver inflammation. We analyzed immune cell 

subsets in organs from experimental groups (WATER pre-ConA, 

WATER-ConA, DSS-pre ConA, and DSS-ConA). RESULTS: In 

DSS-ConA mouse liver, numbers of CD4 + and CD8 + T cells 

were decreased significantly, compared with the WATER-ConA 

group, but numbers of Foxp3 + CD4 + regulatory T cells (Tregs) 

were unchanged. Importantly, gene expression of the inflammatory 

cytokines interferon-gamma (IFN-γ) and tumor necrosis 

factor (TNF) in sorted T cells were significantly decreased in the 

DSS-ConA group. We recently reported that TNF-producing 

CCR9 + CD11b + macrophages were critical in the pathogenesis 

of ConA-induced acute liver injury by inducing T helper 

1 responses. In WATER-ConA-treated livers, CCR9 + CD11b + 

macrophages (WATER-ConA mac) were increased and CCR9 - 

CD11b + macrophages (DSS-ConA mac), a functionally distinct 

subset from WATER-ConA mac, were increased in DSS-ConAtreated 

livers. Sorted DSS-ConA mac had regulatory characteristics 

and produced IL-10, but low levels of TNF or IFN-γ after 

LPS stimulation in vitro. DSS-ConA mac expressed low MHC 

class II and CD80, and antigen presentation to naïve CD4 T 

cells derived from ovalbumin-specific αβ-TCR transgenic mice 

was deficient. CONCLUSIONS: CCR9 - CD11b + macrophages 

migrate to the inflamed liver under a colitic state and contribute 

to immunological tolerance in the liver by suppressing T-cell 

immunity. Our data provide a novel mechanism underlying 

immune tolerance in the liver through the gut-liver axis. 

Disclosures: 

Takanori Kanai - Grant/Research Support: Mitsubishi Tanabe Pharma Corporation 

The following authors have nothing to disclose: Nobuhito Taniki, Nobuhiro Nakamoto, 

Hirotoshi Ebinuma, Po-sung Chu, Takeru Amiya, Shunsuke Shiba, Akihiro 

Yamaguchi, Yuko Wakayama, Hidetsugu Saito


2132 

Small-specific-sized hyaluronic acid prevents inflammation 

and liver injury in chronic ethanol-fed mice 

Damien Bellos 1 , Megan R. McMullen 1 , Rebecca L. McCullough 1 , 

Paramananda Saikia 1 , Sanjoy Roychowdhury 1 , Carol A. de la 

Motte 1 , Laura E. Nagy 1,2 ; 1 Lerner Research Institute, Cleveland 

Clinic, Cleveland, OH; 2 Center for Liver Disease Research, Case 

Western Reserve University, Cleveland, OH 

Chronic alcohol consumption leads to the activation of Kupffer 

cells and remodeling of the extracellular matrix to release hyaluronic 

acid fragments. Classically HA fragments are thought 

to act as DAMPs that promote inflammation; however, growing 

evidence indicates that small-specific-sized HA (SSS-HA) 

downregulate inflammatory signaling. We find that SSS-HA 

normalizes TNFα expression in primary Kupffer cells isolated 

from ethanol-fed rats. To investigate these findings in mice, 

female C57BL/6 mice were fed a Lieber DeCarli liquid diet 

with increasing concentrations of ethanol or pair-fed isocaloric 

maltodextrose over 25 days. SSS-HA was included or 

not in the liquid diet at 300ug/mouse/day. SSS-HA protected 

from ethanol-induced from increases in plasma ALT, AST and 

accumulation of hepatic triglycerides. SSS-HA also normalized 

lipid peroxidation and cell death in the liver as determined 

by 4-HNE and TUNEL staining, respectively. This decrease in 

liver injury was paralleled by lower expression of mRNA for 

inflammatory genes such as TNFα and IL-6. Semi-quantification 

of hepatic TNFα by immunohistochemistry showed that SSS-HA 

also prevented the ethanol-induced increase in TNFα in parenchymal 

and non-parenchymal cells. Taken together, these findings 

indicate that SSS-HA prevents liver injury by preventing the 

ethanol-induced increase in inflammatory signaling. Treatment 

with SSS-HA represents a potential pharmacologic intervention 

that could be targeted to normalize liver inflammation in ALD 

patients. 

Disclosures: 

The following authors have nothing to disclose: Damien Bellos, Megan R. McMullen, 

Rebecca L. McCullough, Paramananda Saikia, Sanjoy Roychowdhury, Carol 

A. de la Motte, Laura E. Nagy 

2133 

Sparstolinin B, a TLR4-antagonist Attenuates Early Steatohepatitic 

Injury in Progressive NonAlcoholic Steatohepatitis 

Diptadip Dattaroy 1 , Ratanesh K. Seth 1 , Suvarthi Das 1 , Firas Alhasson 

1 , Daping Fan 2 , Mitzi Nagarkatti 3 , Prakash Nagarkatti 3 , Saurabh 

Chatterjee 1 ; 1 Environmental Health Sciences, University of 

South Carolina, Columbia, SC; 2 Department of Cell Biology and 

Anatomy, University of South Carolin aSchool of Medicine, Columbia, 

SC; 3 PMI, University of South Carolina Shool of Medicine, 

Columbia, SC 

Nonalcoholic Steatoheaptitis, a hepatic manifestation of metabolic 

syndrome arises from an underlying condition of obesity. 

It is estimated that a roughly 5-8% (500,000) of obese 

population in USA alone have NASH. Sparstolonin B (SsnB) 

is derived from Chinese herb Spaganium stoloniferum and is 

a potent bioactive compound which has already been characterized 

to act as Toll like receptor 4 (TLR4) antagonist. In this 

research we hypothesize that SsnB attenuates early steatohepatitic 

injury and inflammation by inhibiting TLR4-Flotilin colocalization, 

decreased miR21 expression and PTEN activation. To 

prove our hypothesis, we have used an early steatohepatitic 

injury model of diet induced obese (DIO) mice administered 

with bromodichloromethane (BDCM) for 1 week. A similar 

group of mice were administered with SsnB for 1 week along 

with BDCM (SsnB treated group). Quantitative real time PCR 

(qRT PCR) results reveled that there was a significant decrease 

in CD68 (macrophage activation marker), MCP1 (monocyte 

chemoattractant protein 1), IL1β, TNFα, IL23 (inflammatory 

cytokines) gene expression in SsnB treated mice liver compared 

to BDCM treated group. SsnB treatment was found to decrease 

TLR4-flottilin colocalization compared to untreated group-thus 

attenuating TLR4 signaling. Our previous published research 

has shown that Micro RNA 21(miR21) upregulation causes 

inflammation and fibrosis in steatohepatitic injury by inhibiting 

its target proteins. Several other studies also show downregulation 

of PTEN and activation of PI3k/AKT pathway. Results in 

this model showed significantly decreased miR21 expression in 

SsnB treated mice liver compared to only BDCM treated mice. 

Increased PTEN activation is protective as it is a negative regulator 

of PI3K/AKT pathway and thus acts as a protective factor 

against steatohepatitic liver injury. Western blot data confirms 

significantly higher PTEN expression in SsnB treated mice liver 

as compared to BDCM treated mice. Serum ALT, a marker of 

liver injury was also decreased in SsnB treated mice as compared 

to BDCM-treated mice where liver injury was prominent. 

Taken together, we report a novel attenuation of steatohepatitic 

injury function of SsnB. Future studies will focus on molecular 

mechanisms in isolated cells to unravel the binding, kinetics 

and abrogation of TLR4 signaling in hepatic stellate cells. 

Disclosures: 

The following authors have nothing to disclose: Diptadip Dattaroy, Ratanesh 

K. Seth, Suvarthi Das, Firas Alhasson, Daping Fan, Mitzi Nagarkatti, Prakash 

Nagarkatti, Saurabh Chatterjee 

2134 

Macrophage activation through hypoxia inducible factor 

1 alpha (HIF1a) signaling is essential in parenteral 

nutrition associated cholestasis in mice 

Karim C. El Kasmi, Aimee Anderson, Michael W. Devereaux, 


Background: Parenteral Nutrition Associated Cholestasis 

(PNAC) is most severe and progressive in children and adults 

with intestinal failure. We have previously reported that LPS 

absorption into the portal vein and transcription of pro-inflammatory 

cytokines (e.g., IL1β) is increased in hepatic macrophages 

in mice with intestinal injury that receive PN and 

develop PNAC. Furthermore, activation of macrophages and 

suppression of hepatic bile salt export pump Abcb11/BSEP 

and bilirubin exporter Abcc2/MRP2 required intact TLR4 signaling 

in this model. (Sci. Transl. Med. 2013 ;5:206ra137). 

An important role for the transcription factor HIF1a has 

been reported in pro-inflammatory macrophage activation in 

response to LPS-TLR4 signaling and that HIF1 directly increases 

transcription of IL-1b in response to LPS. We therefore hypothesized 

that HIF1α expression in macrophages played a critical 

role in the pathogenesis of PNAC. Methods and Results: PNAC 

model: Wild type C57/B6 mice were maintained on infusion 

of phytosterol-containing (soy lipid) PN solution through a central 

venous catheter for 14 days (DSS-PN mice) following exposure 

to dextran sulfate sodium (DSS) (to induce intestinal injury) 

in drinking water for 4 days. DSS-PN mice at 14 d developed 

cholestasis (increased serum bile acids and bilirubin), 

hepatocyte injury (increased AST and ALT), hepatic macrophage 

hyperplasia and hypertrophy by immunohistochemistry 

(F4/80 stain) and increased expression of IL1b mRNA in liver 

homogenate as well as in isolated intrahepatic macrophages. 

DSS-PN mice displayed significantly reduced hepatic mRNA 

for bile transporters Abcb11 and Abcc2 and the sterol transporter 

(Abcg5/g8). We next used genetic approaches to elucidate 

the roles of IL1b and HIF1a in this model. Global genetic


interruption of IL1b signaling (IL1R -/- mice) prevented PNAC 

in DSS-PN mice, and normalized hepatic macrophages on 

histology and the expression of bile and sterol transporters to 

levels observed in control mice. Macrophage specific genetic 

deletion of HIF1a (Hif1a-LysMcre) in DSS-PN treated mice also 

normalized AST, ALT, bile acids, and bilirubin, whereas similarly 

treated wild type mice showed evidence of hepatocyte 

injury and cholestasis. Moreover, hepatic gene expression of 

Abcb11, Abcc2, and Abcg5/g8 was not reduced in DSS-PN 

HIF1a-LysMcre mice. Conclusions: These data support a key 

role of macrophage activation through gut-derived LPS in triggering 

the development of PNAC. Moreover, these data are 

consistent with HIF1 as a critical upstream regulator of IL1b 

signaling in hepatic macrophages. Finally, blocking HIF1a 

signaling in macrophages is sufficient to prevent PNAC in a 

mouse model. 

Disclosures: 





Michael W. Devereaux 

2135 

Knockdown of Receptor Interacting Protein kinase-1 

(RIPK1) markedly exacerbates immune-mediated liver 

injury and induces lethality through massive TNFα/ 

caspase-dependent apoptosis of hepatocytes, independently 

of RIPK3/MLKL-mediated necroptosis. 

Jo Suda 1 , Lily Dara 1 , Luoluo Yang 1,3 , William A. Gaarde 2 , Neil 

Kaplowitz 1 , Zhang-Xu Liu 1 ; 1 Medicine, University of Southern 

California, Los Angeles, CA; 2 ISIS Pharmaceutics, Carlsbad, CA; 

3 Medicine, Bethune First Hospital of Jilin University, Changchuan, 

China 

Background: RIPK1 has an essential role in the signaling pathways 

triggered by death receptors (DR) through activation of 

NF-kB and regulation of caspase-dependent apoptosis and 

RIPK3/MLKL-mediated necroptosis. The pathophysiological 

roles of RIPK1 and RIPK3 are controversial in immune-mediated 

liver injury. Alpha-galactosylceramide (αGal), a specific activator 

for NKT cells, induces a well-established immune-mediated 

liver injury predominantly through DR pathways. Aim: examine 

the effect of RIPK1 knockdown on αGal-mediated liver injury. 

Methods: C57BL/6 mice received 4μg/mouse of αGal i.p. to 

induce immune-mediated liver injury. Injection of RIPK1 antisense 

(ASO), every other day, for 10 days efficiently silenced 

RIPK1 expression in liver. Results: αGal caused moderate liver 

injury in control ASO-treated mice characterized by small 

patchy inflammatory necrotic foci in histology and mild increase 

in serum ALT (190±110 at 6hr, n=10), with 100% survival. 

However, in RIPK1 ASO-treated (RIPK1-KD) mice, αGal caused 

severe liver injury evidenced by markedly increased serum ALT 

(12,403±3547 at 6hr, n=10, p


TLR9 antagonists into DKO mice ameliorated serum ALT levels 

and the expression levels of IFN-beta. These results suggested 

that the activation of TLR9 was involved in the exacerbation 

of liver injury observed in DKO mice. Conclusion: Deficiency 

of DNase in apoptotic-prone hepatocytes with mitochondrial 

dysfunction is considered to induce necrotic cell death through 

the TL9 signaling pathway. 

Disclosures: 






The following authors have nothing to disclose: Yoshinobu Saito, Yasutoshi 

Nozaki, Yugo Kai, Yuki Makino, Tasuku Nakabori, Satoshi Tanaka, Satoshi 

Aono, Ryotaro Sakamori, Naoki Hiramatsu, Tomohide Tatsumi 

2137 

TRPV4 regulates inflammation and Kupffer cell activation 

in nonalcoholic steatohepatitis by attenuation of 

CYP2E1-mediated oxidative stress 

Ratanesh K. Seth 1 , Suvarthi Das 1 , Diptadip Dattaroy 1 , Firas Alhasson 

1 , Phillip D. Bell 2 , Gregory A. Michelotti 3 , Mitzi Nagarkatti 4 , 

Prakash Nagarkatti 4 , Wolfgang B. Liedtke 5 , Anna Mae Diehl 3 , 

Saurabh Chatterjee 1 ; 1 ENVIRONMENTAL HEALTH SCIENCES, 

UNIVERSITY OF SOUTH CAROLINA, Columbia, SC; 2 Department 

of Medicine, Medical University of South Carolina, Charleston, 

SC; 3 Division of Gastroenterology, Duke University, Durham, NC; 

4 Department of Pathology, Microbiology and Immunology, University 

of South Carolina School of Medicine, Columbia, SC; 5 Department 

of Neurology, Duke University Medical Center, Durham, NC 

Emerging evidence shows that oxidative stress via the activation 

of cytochrome p450 2E1 (CYP2E1) is key to progression 

of inflammation in nonalcoholic steatohepatitis (NASH). We 

have shown previously that CYP2E1 mediated oxidative stress 

and macrophage polarization in NASH livers were attenuated 

by administration of NO donor. However the molecular 

mediator and its pathways that regulate CYP2E1 mediated 

oxidative stress in NASH remains obscure. For this study we 

used a high fat diet induced obese mice (DIO), where CYP2E1 

substrate bromodichloromethane (BDCM) was used to induce 

CYP2E1 mediated oxidative stress, inflammation and NASH 

pathology. Results showed that the transient receptor potential 

vanilloid channel 4 (TRPV4) expression and protein levels were 

significantly elevated in parallel to increases in CYP2E1 and 

correlated well with increased lipid peroxidation, IL-1β, MCP- 

1, TNF-α and HMGB1 levels in the NASH livers and immortalized 

Kupffer cells. TRPV4 knockout (KO) mice exposed with 

BDCM showed increased CYP2E1 protein, lipid peroxidation, 

inflammatory cytokines, infiltration of leukocytes, endothelial 

dysfunction maker genes (CD34, cdh5, ICAM-1 and VEGFR2), 

HMGB1 levels, decreased endothelial nitric oxide synthase 

(NOS3) phosphorylation and exhibited early morbidity as 

compared to wild type mice with NASH. Mechanistically, diallyl 

sulfide (CYP2E1 inhibitor) or NO donor (DETANONOate) 

administration to TRPV4 KO mice completely abrogated 

enhanced NASH symptoms and morbidity. Interestingly, use 

of NO donor significantly decreased expression of CYP2E1, 

CYP2E1-mediated lipid peroxidation, an indirect measure of 

its activity, proinflammatory genes and endothelial dysfunction 

markers in TRPV4 KO mice. The results obtained show that 

TRPV4, a crucial protein responsible for sensing changes in 

osmotic pressure and Ca 2+ also regulates CYP2E1-mediated 

oxidative stress, inflammation and endothelial injury probably 

by activating NOS3 and release of nitric oxide. Based on the 

above, targeting TRPV4 or its downstream signaling cascade 

might be a promising therapeutic strategy in NASH. 

Disclosures: 

The following authors have nothing to disclose: Ratanesh K. Seth, Suvarthi Das, 

Diptadip Dattaroy, Firas Alhasson, Phillip D. Bell, Gregory A. Michelotti, Mitzi 

Nagarkatti, Prakash Nagarkatti, Wolfgang B. Liedtke, Anna Mae Diehl, Saurabh 

Chatterjee 

2138 

Immune Regulation of Mesenchymal Stem Cells Is 

Dependent on Notch/Cox2/PGE2-Mediated Macrophage 

Differentiation in Liver Inflammatory Injury 

Changyong Li, Shi Yue, Ronald W. Busuttil, Jerzy Kupiec-Weglinski, 

Bibo Ke; The Dumont-UCLA Transplant Center, Department 

of Surgery, David Geffen School of Medicine at UCLA, Los Angeles, 

CA 

Background: Mesenchymal stem cells (MSCs) have been 

shown an immune modulatory properties and tissue repairing 

capability under inflammatory conditions. However, it is 

unknown how MSCs may affect immunity in liver inflammatory 

injury. This study was designed to dissect the molecular mechanisms 

of MSCs in regulating immune responses in ischemia and 

reperfusion (IR)-triggered liver inflammation. Methods: Bone 

marrow-derived MSCs were transfected with CRISPR/Cas9-mediated 

Notch1 or Cox2 knockout vector (p-Notch1 KO and 

p-Cox2 KO), and then co-cultured with bone marrow-derived 

macrophages using a transwell system followed by LPS (100 

ng/ml) stimulation. Using liver IRI model, mice (n=6/gr) were 

injected i.v. with 1x10 6 MSCs 24 h prior to liver ischemia. Mice 

were sacrificed after 6 h of reperfusion. Results: The expression 

of Notch1, Cox2, and PGE2 was enhanced in LPS-stimulated 

MSCs. Macrophages co-cultured with MSCs reduced M1 macrophage 

iNOS expression yet strongly augmented the expression 

of arginase-1, an M2 macrophage phenotype, which 

accompanied by increased expression of IL-10/TGF-β. However, 

transfection of p-Notch1 KO in MSCs decreased Cox2 

and PGE2 secretion, as well as reduced macrophage expression 

of arginase-1. Furthermore, knockdown of Cox2 by transfecting 

p-Cox2 KO in MSCs diminished PGE2 production and 

IL-10/TGF-β expression. Unlike in controls, adoptive transfer 

of MSCs in mice ameliorated IR-induced liver damage, which 

accompanied by increased expression of Notch1, Cox2, and 

PGE2 in ischemic livers. Furthermore, macrophages isolated 

from MSCs-treated mice displayed an increased expression of 

arginase-1 than those untreated controls. Conclusion: This study 

demonstrates that MSCs can reprogram host macrophage differentiation 

towards an anti-inflammatory M2 phenotype via a 

Notch/Cox2/PGE2-dependent manner. Our findings provide 

a novel therapeutic potential for the management of IR-triggered 

liver inflammatory injury. 

Disclosures: 

Ronald W. Busuttil - Grant/Research Support: Bayer, Fujisawa, Novartis, Roche/ 

Genentech, NIH 

The following authors have nothing to disclose: Changyong Li, Shi Yue, Jerzy 

Kupiec-Weglinski, Bibo Ke


2139 

The attenuation of inflammatory liver injury by alkaline 

phosphatase is impaired in livers lacking asialoglycoprotein 

receptors: role of the major hepatic trafficking 

system in detoxification processes 

Robert S. Bridge 1 , John Gould 2 , Jacy L. Kubik 1 , Carol A. Casey 2,1 , 

Dean J. Tuma 1 , Benita L. McVicker 2,1 ; 1 Internal Medicine, University 

of Nebraska Medical Center, Omaha, NE; 2 VA Nebraska-Western 

Iowa Health Care System, Omaha, NE 

Alkaline phosphatase (ALP) is a known biomarker during 

inflammatory liver diseases and associated lipopolysaccharide 

(LPS) toxicity. It has been noted that ALP expression during disease 

is a hepatoprotective response against LPS toxicity. Also, 

the therapeutic potential of ALP has been shown to involve LPS 

dephosphorylation and attenuation of liver injury following the 

administration of exogenous intestinal ALP (iALP). However, 

the mechanisms involved in ALP-mediated LPS detoxification 

are not defined. We hypothesize that the functional expression 

of a major hepatic trafficking protein, the asialoglycoprotein 

receptor (ASGPR), is a key factor involved in ALP regulation 

and LPS detoxification. It is known that ASGPRs bind, internalize 

and transport ALP in hepatocytes, yet the contribution 

of ASGPRs in ALP’s protective actions is not known. Here, the 

hepatoprotective potential of ALP was investigated in normal 

wild-type (WT) mice compared to ASGP receptor deficient (RD) 

animals subjected to LPS/D-galactosamine (Gal) liver injury 

with or without iALP treatment. Results: LPS/Gal administration 

resulted in enhanced liver injury in the RD animals shown 

by measures of apoptosis, serum transaminases, altered liver 

architecture and inflammatory parameters (6-fold increase 

(p


2141 

Inflammasome Activation Due to Vinyl Chloride Metabolite 

Exposure in NAFLD Caused by High Fat Diet in Mice. 

Lisanne C. Anders 1 , Adrienne M. Bushau 1 , Anna L. Lang 1 , Gavin 

E. Arteel 1 , Matthew C. Cave 2,1 , Craig J. McClain 2,1 , Juliane I. 

Beier 1 ; 1 Pharmacology and Toxicology, University of Louisville 

health Science Center, Louisville, KY; 2 Medicine, University of Louisville, 


Background. Vinyl chloride (VC), a ubiquitous environmental 

contaminant, ranks 4 th on the ATSDR Hazardous Substances 

Priority List. A major paradigm shift in environmental research 

is to assess the impact of underlying disorders that may modify 

risk. A major health problem in the United States and worldwide 

is non-alcoholic fatty liver disease (NAFLD) due to dietary 

excess. NAFLD, the hepatic manifestation of metabolic complications 

due to obesity, may increase the sensitivity to other 

insults. Indeed, studies by our group and others suggest that 

obesity increases susceptibility to environmental hepatotoxicants 

(e.g., industrial solvents). Recent studies demonstrate a 

critical role of the inflammasome in macrophage activation 

during NAFLD. Inflammasome activation is induced by pathogen-associated 

molecular patterns (“PAMPs”), such as LPS, 

as well as by molecules released from dead or dying cells 

(damage-associated molecular patterns; “DAMPs”). Previously 

we have shown that VC metabolite chloroethanol (ClEtOH) 

exacerbated injury and inflammation leading to necrotic cell 

death in an experimental model of high-fat diet (HFD) induced 

NAFLD. The purpose of the current study was to investigate 

the interaction between NAFLD and VC metabolites in the context 

of inflammasome activation in an experimental model of 

HFD-induced obesity. Methods. Mice, fed a HFD (42% milk 

fat) or low fat control diet (LFD; 13% milk fat) for 10 weeks, 

were administered a bolus dose of ClEtOH or vehicle. Animals 

were sacrificed 0-24 hours after ClEtOH exposure. Plasma and 

tissue samples were harvested for determination of liver injury 

and inflammasome activation. Results. In LFD-fed control mice, 

ClEtOH caused no detectable liver damage, as determined by 

plasma parameters (AST and ALT) and histologic indices of 

damage. In HFD-fed mice, ClEtOH increased HFD-induced liver 

damage, steatosis, hepatocyte ballooning, infiltrating inflammatory 

cells, hepatic expression of proinflammatory cytokines 

and markers of endoplasmic reticulum (ER) stress. VC-metabolite 

induced cell death favors necrosis due to mitochondrial dysfunction 

and ATP depletion in this model. Moreover, in animals 

on a HFD, ClEtOH exacerbated expression of key markers 

involved in inflammasome activation, such as NLRP3 and IL-1β. 

Conclusions. Taken together, chloroethanol (as a surrogate VC 

exposure) can exacerbate liver injury and inflammasome activation 

in HFD-induced NAFLD. This serves as proof-of-concept 

that VC hepatotoxicity may be altered by risk-modifying factors 

such as diet-induced obesity and NAFLD. These data implicate 

exposure to VC as a risk factor in the development of liver disease 

in susceptible populations. 

Disclosures: 








The following authors have nothing to disclose: Lisanne C. Anders, Adrienne M. 

Bushau, Anna L. Lang, Gavin E. Arteel, Juliane I. Beier 

2142 

TNFα mediates the liver:lung axis in alcohol-enhanced 

acute lung injury in mice 

Lauren G. Poole 1 , Veronica L. Massey 1 , Edilson Torres-Gonzalez 2 , 

Keith C. Falkner 2 , Deanna L. Siow 1 , Nikole L. Warner 3 , Robin H. 

Schmidt 1 , Jeffrey D. Ritzenthaler 2 , Jesse Roman 2 , Gavin E. Arteel 1 ; 

1 Pharmacology and Toxicology, University of Louisville, Louisville, 

KY; 2 Medicine, University of Louisville, Louisville, KY; 3 Microbiology 

and Immunology, University of Louisville, Louisville, KY 

Background: It is well known that liver and lung injury can 

occur simultaneously during severe inflammation (e.g. multiple 

organ failure). However, whether these are parallel or interdependent 

(i.e. liver:lung axis) mechanisms is unclear. Previous 

studies have shown that chronic ethanol exposure increases 

the incidence, severity, and mortality of sepsis-induced acute 

lung injury (ALI). There is a known liver:lung axis, and previous 

studies have suggested that hepatic cytokines can contribute to 

pulmonary inflammation; however, this hypothesis in the context 

of alcohol exposure has not been tested. Therefore, the purpose 

of the current study was to investigate the role of hepatic 

cytokine release in alcohol-enhanced lung injury. Methods: 

Male mice were exposed to ethanol-containing Lieber-DeCarli 

diet or pair-fed control diet for 6 weeks. Some animals were 

administered intraperitoneal lipopolysaccharide (LPS) 4 or 24 

hours prior to sacrifice to mimic sepsis-induced ALI. The effect 

of systemic TNFα depletion on lung injury was determined with 

a TNFα-inactivating antibody that is restricted to the plasma 

compartment (etanercept). The expression of cytokine mRNA 

in lung and liver tissue was determined by qPCR. Cytokine 

levels in the bronchoalveolar lavage fluid (BALF) and plasma 

were determined by Luminex assay. Results: The combination 

of ethanol and LPS caused enhanced liver injury, as indicated 

by significantly increased levels of the transaminases ALT/AST 

in the plasma and by changes in liver histology. In the lung, 

ethanol pre-exposure enhanced pulmonary inflammation and 

alveolar hemorrhage caused by LPS. As expected, ethanol 

enhanced LPS-induced TNFα expression in the liver; this effect 

of ethanol was not observed in the lung. In contrast, TNFα-dependent 

chemokines (MIP-2 and KC) were superinduced by the 

combination of ethanol and LPS. Systemic TNFα depletion with 

etanercept almost completely prevented the enhancement of 

lung damage. Furthermore, protecting against this damage correlated 

with an almost complete attenuation of the enhanced 

induction of TNFα-responsive chemokines MIP-2 and KC. Conclusions: 

Chronic ethanol pre-exposure enhanced both liver 

and lung injury caused by LPS. Enhanced organ injury corresponded 

with unique changes in the pro-inflammatory cytokine 

expression profiles in the liver and the lung. Although there 

are also likely pulmonary-specific changes caused by alcohol, 

these data suggest that systemic (hepatic) TNFα drives alcohol-enhanced 

ALI. 

Disclosures: 

Jesse Roman - Advisory Committees or Review Panels: Cellgene; Grant/Research 

Support: Actelion, Intermune, Novartis 

The following authors have nothing to disclose: Lauren G. Poole, Veronica L. 

Massey, Edilson Torres-Gonzalez, Keith C. Falkner, Deanna L. Siow, Nikole L. 

Warner, Robin H. Schmidt, Jeffrey D. Ritzenthaler, Gavin E. Arteel


2143 

Osteopontin promotes cholangiocyte chemokine secretion 

and macrophage accumulation in mice 

Jason D. Coombes 1 , Paul P. Manka 1,2 , Marzena Swiderska-Syn 3 , 

Antonio Riva 4 , Danielle Reid 5 , Lee C. Claridge 6 , Laurent Dollé 7 , 

Rasha Younis 1 , Marco A. Briones-Orta 1 , Naoto Kitamura 1 , Zhiyong 

Mi 8 , Paul C. Kuo 8 , Roger Williams 1,4 , Anna Mae Diehl 3 , 

Shilpa Chokshi 4 , Bertus Eksteen 5 , Ali Canbay 2 , Wing-Kin Syn 1,8 ; 

1 Regeneration and Repair Group, Institute of Hepatology, Foundation 

for Liver Research, London, United Kingdom; 2 Department 

of Gastroenterology and Hepatology, Essen University Hospital, 

Essen, Germany; 3 Department of Medicine, Division of Gastroenterology, 

Duke University, Durham, NC; 4 Viral Hepatitis and 

Alcohol Research Group, The Institute of Hepatology, Foundation 

for Liver Research, London, United Kingdom; 5 Snyder Institute for 

Chronic Diseases, Health Research and Innovation Centre (HRIC), 

University of Calgary, Calgary, AB, Canada; 6 Department of 

Hepatology, Leeds Teaching Hospital NHS Trust, Leeds, United 

Kingdom; 7 Liver Cell Biology Lab (LIVR), Department of Cell Biology 

(CYTO),, Faculty of Medicine and Pharmacy, Vrije Universiteit 

Brussel, Brussels, Belgium; 8 Department of Surgery, Loyola University 

Chicago, Chicago, IL 

Background/Aim: Liver disease progression is characterized 

by recruitment and accumulation of inflammatory cells, which 

cluster around the peri-portal regions (i.e. ductular inflammatory 

response). We recently reported that repair-associated 

Hedgehog (Hh) ligands could induce cholangiocytes to secrete 

chemokines. The downstream Hh target Osteopontin (OPN) 

is widely upregulated during tissue injury and repair, and 

has been shown to regulate macrophage functions via pro-inflammatory 

chemoattractant properties. Although OPN is recognised 

as a key driver of liver fibrogenesis via activation of 

hepatic stellate cells and progenitor cells, the role of OPN in 

liver macrophage activation and recruitment remains unclear. 

We hypothesized that OPN stimulates cholangiocyte production 

of chemokines responsible for recruiting macrophage 

subsets that enhance liver fibrogenesis. Methods: In vivo: the 

role of OPN on total liver chemokine expression and macrophage 

numbers were assessed in two models of liver fibrosis 

(methionine-choline deficient diet and 3, 5,-diethoxycarbonyl-1,4-dihydrocollidine 

diet) by qRTPCR and/or FACS analysis 

(CD11b, Gr1, F4/80, CCR2) of liver infiltrating mononuclear 

cells. In vitro: OPN knockdown was achieved in murine 603B 

cholangiocytes using lentiviral mediated shRNA, and secreted 

OPN neutralized using specific aptamers. To determine if OPN 

modulates cholangiocyte chemokine production, conditioned 

media and cell lysates were harvested. Cytokine secretion was 

measured by cytometric bead array and mRNA by qRTPCR. 

The RAW264.7 cell line was used to study macrophage migration 

in a transwell assay. Results: At the end of treatment, MCD 

and DDC-fed mice developed liver fibrosis, upregulated total 

liver OPN, TGF-β, MCP-1, RANTES, and CXCL1 mRNA, and 

accumulated liver CD11b/F480(+) CCR2 (hi) macrophages. 

Conversely, MCD-fed mice that received OPN-aptamers (which 

neutralizes circulating OPN) downregulated MCP-1, RANTES, 

and CXCL1 mRNA, reduced liver CD11b/F4/80(+) CCR2 (hi) 

by 30%, and developed less fibrosis. In vitro, OPN knockdown 

reduced the secretion of chemokines RANTES, MCP-1 

and CXCL1 (by 75%, 73% and 41%, respectively). There were 

similar reductions in RANTES, MCP-1 and CXCL1 mRNA. Additionally, 

RAW264.7 macrophages cultured with conditioned 

media from 603B treated with OPN-neutralizing aptamer 

exhibited a 35% decrease in migration. Conclusions: OPN is 

overexpressed in progressive liver disease, enhances cholangiocyte 

production of key macrophage chemokines MCP-1, 

RANTES, and CXCL1, and promotes macrophage accumulation. 

OPN-neutralization leads to attenuated fibrogenic outcomes 

and is a promising anti-fibrotic strategy. 

Disclosures: 

The following authors have nothing to disclose: Jason D. Coombes, Paul P. 

Manka, Marzena Swiderska-Syn, Antonio Riva, Danielle Reid, Lee C. Claridge, 

Laurent Dollé, Rasha Younis, Marco A. Briones-Orta, Naoto Kitamura, Zhiyong 

Mi, Paul C. Kuo, Roger Williams, Anna Mae Diehl, Shilpa Chokshi, Bertus 

Eksteen, Ali Canbay, Wing-Kin Syn 

2144 

Liver Enzymes Elevation In The Setting Of Chronic 

Graft-Versus-Host Disease Is A Non Specific Marker Of 

Inflammation That Does Not Accurately Predict Disease 

Related Hepatic Injury 

Ma Ai Thanda Han 4 , Niharika Samala 4 , Bisharah S. Rizvi 2 , Kenneth 

J. Wilkins 4 , Ohad Etzion 4 , Elizabeth C. Jones 5 , Christopher 

Koh 4 , David E. Kleiner 1 , Steven Pavletic 3 , Theo Heller 4 ; 1 LABORA- 

TORY OF PATHOLOGY, NCI, Bethesda, MD; 2 Internal Medicine, 

Canton medical education foundation, Canton, OH; 3 NIH-NCI, 

Bethesda, MD; 4 Liver Diseases Branch, NIH/NIDDK, Bethesda, 

MD; 5 Radiology, NIH-Clinical Center, Bethesda, MD 

Introduction: Diagnosis of chronic hepatic graft-versus-host disease 

(HcGVHD) is challenging, as the current scoring system 

(the NIH Scoring System or NSS) is based on total bilirubin 

(TB), alanine aminotransferase (ALT) and alkaline phosphatase 

(ALP). The challenge is that liver associated enzyme (LAE) and 

bilirubin elevations may be nonspecific. Once diagnosed with 

HcGVHD, patients are treated with immunosuppressive medications 

that have risk. Aims: To stratify patterns of LAE that could 

mimic HcGVHD, to assess for cytokine associations, and to 

assess the accuracy of the NSS for HcGVHD. Method: Patients 

with stable chronic GVHD (cGVHD) at any site were enrolled at 

variable times from bone marrow transplant. LAE, PT, TB, albumin, 

platelets, inflammatory markers (ESR, ferritin, C3), and 

cytokines were measured. Patients underwent liver biopsy at 

the physicians’ discretion. Box-Cox power transformation and t 

test were used for testing strength of associations. Results: 302 

patients (58% men, median age 43 y, ALT=39 IU/ml, ALP=96 

IU/ml, platelet=245 x 10 3 /ml) with cGVHD were included. 

.Abnormal ALT and AST were significantly associated with 

lower platelets, and higher TB and PT, in addition to higher ferritin, 

C3, MDC, TGFα, IFNα, IL15 and amyloid A (p= 0.0009, 

0.02, 0.007, 0.02, 0.04, 0.02, 0.04 respectively). Abnormal 

γ glutamyl transferase (GGT) was associated higher TB and 

PT but not with lower platelets. In addition, higher GGT was 

associated with elevated ESR (p=0.04) and C3 (p=0.0002), 

as well as with MDC, TGFα, IL 6, IP10 and IFNγ (p= 0.009, 

0.02, 0.008, 0.04, 0.02 respectively). Abnormal ALT, AST 

and GGT together were associated with abnormal platelet, 

TB, PT, C3 and ferritin (p=0.03, 0.01, 0.0003, 0.0007, 

0.02 respectively), as well as with MDC, and IL15 (p=0.008, 

0.01 respectively). 151/302 patients were diagnosed with 

HcGVHD by NSS. 24/302 patients underwent liver biopsy. 

Among them, 12 patients were diagnosed with HcGVHD by 

NSS of whom only 5 had histologic features that were consistent 

with HcGVHD, while others had characteristics of non-alcoholic 

steatohepatitis or nodular regenerative hyperplasia. 

Of the non-HcGVHD group by NSS, 4 had HcGVHD. Overall 

sensitivity and specificity for HcGVHD was 55% and 53%. 

There was no statistical differences between biopsy proven 

HcGVHD versus non-HcGVHD for AST, ALT, ALP, GGT, platelet, 

TB and albumin. Conclusion: Abnormal LAE in cGVHD is a 

marker of inflammation rather than a reflection of any specific 

pattern of liver injury. The NSS is neither sensitive nor specific 

for HcGVHD diagnosis. Hepatic histology may therefore be


crucial to distinguish the causes of hepatic inflammation in 

cGVHD. 

Disclosures: 

The following authors have nothing to disclose: Ma Ai Thanda Han, Niharika 

Samala, Bisharah S. Rizvi, Kenneth J. Wilkins, Ohad Etzion, Elizabeth C. Jones, 

Christopher Koh, David E. Kleiner, Steven Pavletic, Theo Heller 

2145 

The hepatic and extra-hepatic profile of resolution of 

steatohepatitis induced by GFT-505 

Arun J. Sanyal 2 , Stephen A. Harrison 3 , Sven M. Francque 4 , Pierre 




Alice Roudot 13 , Sophie Megnien 13 , Bart Staels 14 , Vlad Ratziu 1 ; 

1 Hepatology, Hopital Pitie Salpetriere, Paris, France; 2 Internal 

Medicine/Division of Gastroenterology, Hepatology and Nutrition, 

Virginia Commonwealth University, Richmond, VA; 3 Gastroenterology, 

Brooke Army Medical Center, Fort Sam Houston, 

TX; 4 Gastroenterology Hepatology, Antwerp Univesrity Hospital, 

Antwerp, Belgium; 5 Pathology, Hopital Beaujon, Clichy, France; 

6 Hepatology Department, Saint-Antoine Hospital, Paris, France; 

7 Digestive Diseases, Valme University Hospital, Sevilla, Spain; 

8 Hepatology Department, University Hospital & LUNAM University, 

Angers, France; 9 Medicine, Duke University, Durham, NC; 

10 Hepatology Department, University of Virginia, Charlotesville, 

VA; 11 Department of Gastroenterology and Hepatology, RUNMC, 

Nijmegen, Netherlands; 12 Institute of cellular Medicine, Newcastle 

University, Newcastle, United Kingdom; 13 Genfit SA, Loos, 

France; 14 INSERM U1011, European Genomic Institute for Diabetes 

(EGID), Institut Pasteur de Lille, Lille, France 

Background: Non-alcoholic steatohepatitis (NASH) is associated 

with increased liver- and cardio-metabolic risks. It is 

unknown whether resolution of steatohepatitis (SH) is associated 

with fibrosis regression and changes in cardio-metabolic 

risk profile, and whether drug-induced resolution of SH is similar 

to spontaneous resolution of SH. AIMS: We compared the 

hepatic and extra-hepatic profile associated with resolution 

of SH in GOLDEN505, a phase 2 randomized placebo-controlled 

trial of GFT505, a PPAR α/δ agonist. METHODS: Subjects 

with SH and NAS≥4 who received GFT505 120 mg/ 

day (G120) or placebo (PBO) were studied. SH was defined 

by the presence of steatosis, inflammation and hepatocyte ballooning. 

Resolution of SH was defined by loss of any one of 

these components. Responders (R) were defined as resolution 

of SH without worsening of fibrosis. R and non-responders 

(NR) to G120 were compared for hepatic and extra-hepatic 

activity, and G120 R were compared to PBO R to explore the 

drug-induced effect of reversal of SH on extra-hepatic improvements. 

RESULTS: 202 subjects were included. The proportion of 

R was greater in G120 vs. PBO (22.4% vs. 12.7%; RR 2.01, 

p


(advanced NASH: a-NASH)). 13 C-labeled palmitate (an SFA) 

was administered directly into the duodenum using gastrointestinal 

endoscopy to avoid delays resulting from delivery by 

stomach. Breath 13 CO 2 

levels were then measured to quantify 

metabolized SFA, and apolipoprotein B-48 concentrations in 

postprandial serum after test meal were measured to quantify 

absorbed chylomicrons in the intestine. Expression of SFA 

transporters in intestinal specimens from these groups was also 

examined. Results: Overall, 13 CO 2 

excretion was significantly 

higher in the e-NASH group than in the control and a-NASH 

groups (P


centers (Hôpital Beaujon, Clichy and Hospital Clinic, Barcelona). 

Plasma samples were obtained from peripheral blood 

and M65, M30, CCL20 and TREM1 levels were measured by 

ELISA. The fraction of M65 and M30 carried by microparticles 

(MP) was determined. Plasma levels of biomarkers were compared 

between patients with and without histological diagnosis 

of AH. RESULTS AH was histologically confirmed in 47 of 88 

patients from the test cohort. The ABIC score was B or C in 

71/88 patients (81%), and 40/47 (85%) in those with AH. 

Patients with AH had higher median plasma levels of circulating 

free and MP-bound M65 (8.0 and 12.2-fold), free and 

MP-bound M30 (9.4 and 21.6-fold) and CCL20 (4.0-fold) than 

patients without AH (p


2150 

Cross-sectional and longitudinal agreement of magnetic 

resonance imaging proton density fat fraction with 

pathologist grading of hepatic steatosis in adults with 

nonalcoholic steatohepatitis in a multi-center trial 

Michael S. Middleton 1 , Elhamy Heba 1 , Catherine A. Hooker 1 , 

Brent A. Neuschwander-Tetri 2 , Mustafa R. Bashir 3 , Kathryn 

Fowler 4 , Kumar E. Sandrasegaran 11 , Elizabeth M. Brunt 5 , David 

E. Kleiner 6 , Edward Doo 7 , Mark L. Van Natta 8 , James Tonascia 9 , 

Rohit Loomba 10 , Claude B. Sirlin 1 ; 1 Radiology, UCSD, San Diego, 

CA; 2 Medicine, Saint Louis University, St. Louis, MO; 3 Radiology, 

Duke University, Durham, NC; 4 Radiology, Washington University, 

St. Louis, MO; 5 Pathology and Immunology, Washington University, 

St. Louis, MO; 6 Center for Cancer Research, NIH, Bethesda, 

MD; 7 Liver Disease Research Branch, NIH, Bethesda, MD; 8 Epidemiology, 

Johns Hopkins University, Baltimore, MD; 9 Biostatistics 

and Epidemiology, Johns Hopkins University, Baltimore, MD; 

10 Medicine, UCSD, San Diego, CA; 11 Radiology, Indiana University, 

Indianapolis, IN 

Materials and Methods Magnetic resonance imaging (MRI) 

was offered at baseline and end of treatment (EOT) to adults 

participating in the Farnesoid X Receptor Ligand Obeticholic 

Acid in NASH Treatment (FLINT) trial. Proton density fat fraction 

(PDFF), a non-invasive MRI biomarker for hepatic steatosis, 

was estimated using an advanced gradient-recalled-echo 

sequence that avoids or corrects confounding factors that can 

cause hepatic fat quantification to be inaccurate or scanner 

dependent. Imaging quality control was managed centrally 

by the NASH CRN Radiology Coordinating Center. Histologic 

steatosis grade, scored centrally by the NASH CRN Pathology 

Committee, served as the reference standard for PDFF. 

Cross-validated receiver operating characteristic (ROC) analyses 

were performed. Results Of 283 adults enrolled in FLINT, 

113 had MRI and biopsy at baseline, 85 had MRI and biopsy 

at EOT, and 78 had MRI and biopsy at both time points. MRIs 

were obtained at seven clinics using MRI scanners from three 

manufacturers, operating at one of two field strengths. Timing 

of MRIs ranged from 29 days before to 89 days after 

biopsy. At baseline, 34% of biopsies had steatosis grade 0 

or 1, 39% had grade 2, and 27% had grade 3 with corresponding 

mean(SD) PDFF(%) of 9.8(3.7), 18.1(4.3), and 

30.1(8.1), respectively. The area under ROC (AUROC) from 

logistic regression using PDFF as a surrogate for classifying 

steatosis grade 2+ vs. < 2 was 0.95 (95% CI: 0.91, 0.98) and 

for classifying steatosis grade 3 vs. < 3 was 0.96 (95 % CI: 

0.93, 0.99). PDFF cut-off values at 90% specificity were 16.3% 

for grade 2+ and 21.7% for grade 3 discrimination; sensitivities 

were 83% and 84%, respectively. At EOT, 42% of paired 

biopsies had improvement in steatosis grade, 49% had no 

change, and 9% had worsening with corresponding mean(SD) 

change from baseline in PDFF(%) of -7.4(8.7), 0.3(6.3) and 

7.7(6.0), respectively. The AUROC using PDFF change to classify 

steatosis grade improvement and worsening, respectively, 

were 0.81 (95% CI: 0.71, 0.91) and 0.81 (95% CI: 0.63, 

0.99). Cut-off values for PDFF change at 90% specificity were 

-5.1% for improvement and +5.6% for worsening; sensitivities 

were 58% and 57%, respectively. Conclusions In a multi-center 

setting, MRI PDFF showed high agreement with histologic 

hepatic steatosis grades on scanners at different sites using 

different scanner manufacturers and of different field strengths. 

Importantly, change in PDFF accurately classified change in 

72-week histologic hepatic steatosis grade. These findings support 

wider use of MRI PDFF in multi-center trials as a biomarker 

for hepatic steatosis at baseline, and for change in hepatic 

steatosis with treatment. 

Disclosures: 

Michael S. Middleton - Consulting: Bracco; Grant/Research Support: Gilead, 

Isis, Genzyme, Pfizer, Siemens, Bayer, Synageva, Merck, Janssen; Stock Shareholder: 

General Electric 



Scholar Rock 

Elizabeth M. Brunt - Consulting: Synageva; Independent Contractor: Rottapharm 







The following authors have nothing to disclose: Elhamy Heba, Catherine A. 

Hooker, Mustafa R. Bashir, Kathryn Fowler, Kumar E. Sandrasegaran, David E. 

Kleiner, Edward Doo, Mark L. Van Natta, James Tonascia 

2151 

Non-invasive Diagnosis of Non-alcoholic Steatohepatitis 

(NASH) using a Panel of Fatty Acids 

Donjeta Gjuka 1 , Xiaoling Song 2 , Wonbeak Yoo 1 , Suk Young 

Yoo 3 , Jing Wang 3 , Michael B. Fallon 4 , George N. Ioannou 5 , Stephen 

A. Harrison 6 , Laura Beretta 1 ; 1 Molecular and Cellular Oncology, 

MD Anderson Cancer Center, Houston, TX; 2 Fred Hutchinson 

Cancer Research Center, Seattle, WA; 3 Bioinformatics and Computational 

Biology, The University of Texas MD Anderson Cancer 

Center, Houston, TX; 4 Division of Gastroenterology, The University 

of Texas Health Science Center at Houston, Houston, TX; 5 Division 

of Gastroenterology, Veterans Affairs Puget Sound Health Care 

System and University of Washington, Seattle, WA; 6 Department 

of Medicine, Brooke Army Medical Center, Houston, TX 

Introduction: Non-alcoholic fatty liver disease (NAFLD) is rapidly 

becoming the most common form of liver disease worldwide. 

NAFLD represents a spectrum of disease states ranging 

from isolated steatosis to non-alcoholic steatohepatitis (NASH). 

Diagnosis of NASH is important as this form of the disease 

has been shown to progress to cirrhosis and hepatocellular 

carcinoma. To date, liver biopsy is required for the diagnosis 

of NASH. Methods: We measured 46 fatty acids in sera from 

106 patients with NAFLD who underwent biopsy at Brooke Military 

Hospital in San Antonio (n=75) and the Veterans Affairs 

Puget Sound Health Care System in Seattle (n=31). For fatty 

acid profiling, total lipids were extracted from 100 μL of serum, 

the phospholipid fraction was isolated by one-dimensional 

thin-layer chromatography and methyl esters of phospholipid 

fatty acids were prepared using direct transesterification and 

separated by gas chromatography. Results: Levels of 15:0, 

17:0 and 16:1n7t negatively correlated with NAFLD activity 

scores (NAS) and hepatocyte ballooning scores, while levels 

of 18:1n7c strongly correlated with fibrosis scores and significantly 

discriminated patients with mild fibrosis and patients 

with intermediate or advanced fibrosis. Levels of 18:1n7c also 

correlated with liver inflammation. In addition, 15:0 and 17:0 

levels negatively correlated with fasting glucose and AST, while 

levels of 16:1n7c, 18:1n7c and 22:5n3 positively correlated 

with AST, ferritin and albumin, respectively. None of the six 

fatty acids correlated with BMI. Inclusion of age, ferritin, AST 

or AST-to-Platelet ratio Index (APRI) scores improved the performance 

of the fatty acid panels in detecting patients with NAS 

≥5 or patients with intermediate to severe fibrosis. The panel 

composed of 15:0, 16:1n7t, 18:1n7c, 22:5n3, age, ferritin 

and APRI best predicted intermediate or advanced fibrosis in 

NAFLD patients, with an area under ROC curve (AUROC) of 

0.92 and a 82% sensitivity at 90% specificity. Conclusion: The 

comprehensive analysis of fatty acid composition through the 

different stages of NAFLD and their correlation to histological


hallmarks of NASH described in this study have allowed for 

the identification of 6 circulating fatty acids that could serve 

as non-invasive markers of disease severity in NAFLD patients. 

It also provides potential insights into mechanisms of disease 

progression. This panel should be further evaluated in prospective 

cohorts for its utility in distinguishing histological severity 

in patients with NAFLD. In addition, the biological effects of 

the fatty acids identified in this study should be investigated to 

evaluate their potential therapeutic utility. 

Disclosures: 

Michael B. Fallon - Grant/Research Support: Bayer/Onyx, Eaisi, Gilead, Grifolis 




The following authors have nothing to disclose: Donjeta Gjuka, Xiaoling Song, 

Wonbeak Yoo, Suk Young Yoo, Jing Wang, George N. Ioannou, Laura Beretta 

2152 

1 H-Magnetic Resonance Spectroscopy is superior to 

Controlled Attenuation Parameter (CAP) in assessing 

liver fat content in human non-alcoholic fatty liver disease 

(NAFLD) 

Jurgen H. Runge 1 , Loek Smits 4 , Joanne Verheij 3 , Aart Nederveen 1 , 

Ulrich Beuers 2 , Jaap Stoker 1 ; 1 Radiology, Academic Medical Center, 

Amsterdam, Netherlands; 2 Gastroenterology and Hepatology, 

Academic Medical Center, Amsterdam, Netherlands; 3 Pathology, 

Academic Medical Center, Amsterdam, Netherlands; 4 Vascular 

Medicine, Academic Medical Center, Amsterdam, Netherlands 

PURPOSE Non-alcoholic fatty liver disease (NAFLD) is an 

increasingly recognized global health problem. Liver biopsy 

is the diagnostic standard, but given its drawbacks the liver 

fat content is preferably assessed noninvasively. The FibroScan® 

now provides the Controlled Attenuation Parameter 

(CAP) as a noninvasive measure of the liver fat content. However, 

only limited data are available regarding its accuracy 

and reproducibility compared to established and validated 

quantitative measures such as H-Magnetic Resonance Spectroscopy 

(H-MRS). Therefore, we prospectively compared 

CAP and H-MRS against liver biopsy in a cohort of NAFLD 

patients. PATIENTS AND METHODS Forty-four NAFLD patients 

(M/F: 33/11) with median (IQR) age of 52.3 (43.8–56.5) 

years and BMI of 27.2 (25.4–33.1) kg/m 2 were included in 

this prospective board-approved study. Same-day 3T MRI and 

CAP measurement were performed within 28 (17–50) days of 

biopsy, the latter assessed by a single hepatopathologist. Within-weeks 

and within-test reproducibility were assessed for CAP/ 

MRI and CAP-only in a subgroup using Bland-Altman limits of 

agreement (BALA). H-MRS fat fractions (FF) and CAP values 

were compared between Brunt steatosis grades S0–S3 using 

Kruskall-Wallis and Mann-Whitney-U tests. Correlations were 

assessed with Spearman’s and diagnostic accuracies of CAP 

and FF to identify ≥S1 on biopsy with ROC analyses. RESULTS 

CAP showed considerable overlap between steatosis grades 

(see Table 1) and only differed between S0 and S2 (p=0.015) 

and S1 and S2 (p=0.004). FF differed (p


length appears to be an important factor impacting histological 

assessments and has important implications for the interpretation 

of placebo-controlled trials in NASH. Shorter biopsy 

length can increase placebo response in a trial and therefore, 

biopsy length should be closely monitored and factored into 

sample-size assessment. 

Disclosures: 







Gilead 






Scholar Rock 














The following authors have nothing to disclose: Xiaohong Yan, Arthur J. 

McCullough 

2154 

Predictors of improvement in NAFLD Activity Score to 

obeticholic acid: A secondary analysis of FLINT Trial 

Rohit Loomba 1 , Arun J. Sanyal 2 , Kris V. Kowdley 3 , Norah Terrault 

4 , Naga P. Chalasani 5 , Manal F. Abdelmalek 6 , Arthur J. 

McCullough 7 , Xiaohong Yan 8 , Reshma Shringarpure 8 , Beatrice 

Ferguson 8 , David Shapiro 8 , Brent A. Neuschwander-Tetri 9 ; 1 University 

of California, San Diego, La Jolla, CA; 2 Virginia Commonwealth 

University, Richmond, VA; 3 Swedish Medical Center, 

Seattle, WA; 4 University of California, San Francisco, San Francisco, 

CA; 5 Indiana University, Indianapolis, IN; 6 Duke University, 

Durham, NC; 7 Cleveland Clinic, Cleveland, OH; 8 Intercept 

Pharmaceuticals, San Diego, CA; 9 Saint Louis University Health 

Sciences Center, St. Louis, MO 

Background: The Farnesoid X Receptor (FXR) Ligand Obeticholic 

Acid (OCA) in NASH Treatment (FLINT) Trial showed 

that OCA led to significantly higher rates of histologic response 

(defined as an improvement in NAFLD activity score [NAS] by 

≥2 with no worsening of fibrosis). Aim: The aims of this secondary 

analysis of the FLINT trial was to identify predictors of histologic 

response after 72 weeks of OCA treatment. Methods: 

Logistic regression model with stepwise selection procedure 

was performed to identify potential predictors of response after 

72 weeks of OCA treatment. Model selection was based on 

Akaike information criterion (AIC) and Bayesian information 

criterion (BIC). All patients included in analysis had biopsies 

at baseline (BL) and 72 weeks. Predictor data collected at BL, 

weeks 12, 24, and 36 were evaluated in this model. 59 predictors 

were assessed including BL values for demographics, 

histology, liver and serum biochemistry as well as the changes 

in those parameters over the course of the trial. The selected 

parameters were used to build a predictive model in OCAtreated 

patients for predicting response defined as improvement 

in the NAS by ≥2 with no worsening of fibrosis. The model was 

cross-validated by jackknife method, area under the receiver 

operating characteristic curve (AUROC), and 95% CIs. Results: 

Among 102 OCA-treated patients, 50 (49%) showed ≥2 point 

improvement in NAS without worsening fibrosis. This predictive 

model showed that older age, higher BL NAS, lower BL BMI, 

higher BL uric acid, and decrease in AST at week 24, increase 

in creatinine at week 24 and greater weight loss at week 24 

was associated with histologic response on OCA treatment 

(Table). The predictive model for placebo-treated patients was 

distinct from the OCA-treated patients (not shown). Conclusions: 

Baseline characteristics and changes in clinical parameters 

may be helpful in predicting histological response to OCA 

therapy in adults with NASH and further evaluation with larger 

patient numbers is warranted. 

Disclosures: 














Gilead 










Beatrice Ferguson - Employment: Intercept Pharmaceuticals 





Scholar Rock 

The following authors have nothing to disclose: Arthur J. McCullough, Xiaohong 

Yan


2155 

Short Sleep Duration is associated with Nonalcoholic 

Fatty Liver Disease in US Adults 

Donghee Kim 1 , Hwa Jung Kim 2 , Alina M. Allen 3 , Aijaz Ahmed 1 , 

Nae-Yun Heo 1 , Prowpanga Udompap 1 , Ajitha Mannalithara 1 , W. 

Ray Kim 1 ; 1 Division of Gastroenterology and Hepatology, Stanford 

University School of Medicine, Stanford, CA; 2 Department 

of Clinical Epidemiology and Biostatistics, Asan Medical Center, 

Seoul, Korea (the Republic of); 3 Division of Gastroenterology and 

Hepatology, Mayo Clinic, Rochester, MN 

Background/Aims: Short sleep duration has been associated 

with increased risk of hypertension, type 2 diabetes and obesity. 

We investigate association between duration and quality 

of sleep and nonalcoholic fatty liver disease (NAFLD). Methods: 

Among adult participants in the National Health and 

Nutrition Examination Survey in 2005-2012, subjects with 

a potential diagnosis of NAFLD were selected by excluding 

individuals with excessive alcohol consumption (> 30 g/day 

for men and 20 g/day for women) or with evidence of viral 

hepatitis. Suspected NAFLD (sNAFLD) was diagnosed if serum 

alanine aminotransferase was >30 U/L for men and >19 U/L 

for women in the absence of other causes. Predicted NAFLD 

(pNAFLD) was diagnosed using fatty liver index (FLI) of ≥ 60 

(based on body mass index (BMI), waist circumference, triglycerides 

and gamma glutamyl transpeptidase). Duration of 

sleep, categorized into ≤ 5, 6, 7, 8, and ≥ 9 hours per night 

and sleep quality, graded as good, moderate and poor, were 

assessed using the Sleep Disorders’ Questionnaire. Results: 

There were 17,245 participants (mean age 46 years, 48.7% 

male) represented in the data set. The most common duration 

of sleep was 7 hours (27.1%), followed by 8 hours (26.3%), 

6 hours (23.8%), ≤ 5 hour (15.9%), and ≥ 9 hour (7.0%). 

Respondents rated their quality of sleep to be good (78.0%), 

moderate (11.7%), or poor (10.3%). The prevalence of sNA- 

FLD was 37.3%, which was inversely correlated with sleep 

duration: 39.2% of respondents with sleep ≤ 5 hours had sNA- 

FLD, 38.0% with 6 hours, 37.3% with 7 hours, 36.6% with 8 

hours, and 33.3% with ≥ 9 hours of sleep (p


larger generalized population. Conclusions: The combination 

of serum Fuc-Hpt and Mac2bp can distinguish NASH from 

NAFLD patients. Our noninvasive model using serum two glycobiomarkers 

contribute to a novel NASH diagnostic methodology 

instead of liver biopsy. 

Disclosures: 



















The following authors have nothing to disclose: Yoshihiro Kamada, Masafumi 

Ono, Hideyuki Hyogo, Hideki Fujii, Yoshio Sumida, Kohjiroh Mori, Saiyu 

Tanaka, Makoto Yamada, Maaya Akita, Kayo Mizutani, Hironobu Fujii, Akiko 

Yamamoto, Shinji Takamatsu, Yuichi Yoshida, Toshiji Saibara, Eiji Miyoshi 

2157 

Increased Immune Responses of Patients with Lean 

Non-Alcoholic Fatty Liver Diseases Over Obese Non-Alcoholic 

Fatty Liver Diseases in Response to Saturated 

Fatty Acid 

Ayub Al Mamun 2 , Mamun A. Mahtab 2 , Sheikh Mohammad Fazle 

Akbar 1 ; 1 Department of Medical Sciences, Toshiba General Hospital, 

Tokyo, Japan; 2 Department of Hepatology, Bangbandhu 

Sheikh Mujib Medical University, Dhaka, Bangladesh 

Purpose: To develop insights about mechanism of pathogenesis 

of obesity in lean persons, this study was initiated to dissect 

immunological events in lean patients with non-alcoholic fatty 

liver diseases (NAFLD) versus obese patients with NAFLD. Methods: 

Out of total 450 patients with NAFLD, clinical parameters 

of age and sex-matched 50 patients with lean NAFLD (body 

mass index (body mass index, BMI) 30.0) were compared. Peripheral 

blood mononuclear cells (PBMC) and antigen-presenting dendritic 

cells (DC) from both lean NAFLD and obese NAFLD 

patients were isolated and production of pro-inflammatory cytokines 

and nitric oxide (NO) in response to various stimuli were 

evaluated. Results: As mentioned, the BMI of lean NAFLD patents 

were significantly lower than that of obese NAFLD patients 

(p0.05). The levels of proinflammatory cytokines [interferon 

(IFN)-gamma, tumor necrosis factor (TNF)-alpha, and interleukin 

(IL)-6] produced by PBMC and DC were also comparable 

between lean NAFLD and obese NAFLD patients when these 

were stimulated by polyclonal stimulators like concanavalin 

A or lipopolysachcharides (p>0.05). However, significantly 

higher levels of IFN-gamma, TNF-alpha, and IL-6 were produced 

by PBMC and DC of lean NAFLD patients compared to 

those produced by obese NAFLD patients in response to palmitic 

acid (p0.05). 

This outcome was reproducible in three separate experiments 

with sera collected at three separate points. Also, nitric oxide 

(NO) production was significantly higher from PBMC of lean 

NAFLD patients compared to obese NAFLD patients (p


DM-HCC individuals than DM-non-HCC individuals (OR 3.44, 

p = 0.0002). The influence of PNPLA3 and JAZF1 was independent 

of age, sex, and body mass index after multivariable 

logistic regression analysis. There were no differences in the 

allele frequencies of PNPLA3 and JAZF1 between the HCV- 

HCC and HCV-non-HCC groups. Conclusions: The PNPLA3 

rs738409 was determined to be associated with HCC development 

in non-viral hepatitis patients with T2DM. Additionally, 

the JAZF1 rs864745 was identified as a risk factor for 

HCC among T2DM patients with the GG genotype at PNPLA3 

rs738409. 

Disclosures: 

The following authors have nothing to disclose: Misuzu Ueyama, Masaaki 

Korenaga, Nao Nishida, Keiko Korenaga, Erina Kumagai, Masaya Sugiyama, 

Yoshihiko Aoki, Masatoshi Imamura, Kazumoto Murata, Naohiko Masaki, 

Takumi Kawaguchi, Takuji Torimura, Hideyuki Hyogo, Hiroshi Aikata, Kiyoaki 

Ito, Yoshio Sumida, Tatsuya Kanto, Sumio Watanabe, Masashi Mizokami 

2159 

Older Age, Increasing Body Mass Index, and Concurrent 

Diabetes Mellitus Increases Risk of Advanced Fibrosis 

Among Nonalcoholic Fatty Liver Disease Patients 

Maria Aguilar 2 , Edward W. Holt 3 , Taft Bhuket 1 , Benny Liu 1 , Zobair 

M. Younossi 4,5 , Aijaz Ahmed 6 , Robert J. Wong 1 ; 1 Gastroenterology 

and Hepatology, Alameda Health System - Highland Hospital, 

Oakland, CA; 2 Medicine, Alameda Health System - Highland 

Hospital, Oakland, CA; 3 Transplantation - Division of Hepatology, 

California Pacific Medical Center, San Francisco, CA; 4 Medicine 

- Center for Liver Diseases, Inova Fairfax Hospital, Falls Church, 


Health System, Falls Church, VA; 6 Division of Gastroenterology 

and Hepatology, Stanford University School of Medicine, Stanford, 

CA 

Background:Nonalcoholic fatty liver disease (NAFLD) is the 

leading cause of chronic liver disease (CLD) in the U.S. and 

will become the leading etiology of hepatocellular carcinoma 

and liver transplantation. However, the true prevalence of 

NAFLD and NAFLD with advanced fibrosis (NAFLD-AF) is not 

well understood. Aim:To determine the current prevalence and 

predictors of NAFLD and NAFLD-AF among U.S. adults. Methods:Using 

the most recent 2011-2012 U.S. National Health 

and Nutrition Examination Survey, we defined NAFLD as presence 

of both abnormal alanine aminotransferase (ALT>20 U/L 

in women, ALT>30 U/L in men) and metabolic syndrome after 

excluding other CLD etiologies (eg. hepatitis C, hepatitis B, 

alcohol). NAFLD-AF was defined as NAFLD fibrosis score > 

0.676. Prevalence estimates were stratified by sex, ethnicity, 

and age. Multivariate logistic regression models evaluated predictors 

of NAFLD and NAFLD-AF. Results:Overall NAFLD prevalence 

was 21.1%, representing 49.2 million persons. NAFLD 

prevalence was higher in females compared to males (31.4% 

vs. 14.0%, p60: 48.2% vs. age


regression analysis of biomarkers indicated that HA, CK18 

and TIMP1 were independent predictor of advanced fibrosis 

(odds ratio [OR] HA=1.057, CK18=1.002, TIMP1=0.99, 

95% CI, p


hypertension, fasting plasma insulin and vitamin B12. In contrast, 

the fibrosis score was higher by 1.4 units in the high 

folate group (RBC folate > 620 ng/ml) compared to low folate 

group (p=0.003) in patients with the homozygous group. The 

fibrosis score increases by 0.06 units for every 25mg increase 

in serum folate level in homozygous group (p=0.014). Plasma 

vitamin B12 was not associated with histological severity. Conclusions. 

While folate has a protective effect on ballooning and 

the overall NAS score in NAFLD, in those NAFLD patients with 

the MTHFR homozygous mutation high folate levels had more 

advanced fibrosis. Despite the reported decreased activity of 

mutated MTHFR, folate supplementation may contribute to progression 

of fibrosis and should be used with caution. 

Disclosures: 

The following authors have nothing to disclose: Jaividhya Dasarathy, Rony Varghese, 

Iryna Kalinina, Rocio Lopez, Arthur J. McCullough, Srinivasan Dasarathy 

2163 

The Lipidomic Signature Of Disease Progression In Nonalcoholic 


Cristina Alonso 1 , Amon Asgharpour 2 , Ibon Martinez-Arranz 1 , 

Puneet Puri 2 , Mohammad S. Siddiqui 2 , Pablo Ortiz 1 , Jose M. 

Mato 3 , Arun J. Sanyal 2 ; 1 OWL, Derio, Spain; 2 VCU Medical Center 

Richmond, Division of Gastroenterology and Hepatology, Richmond, 

VA; 3 CIC bioGUNE, Ciberehd, Derio, Spain 

BACKGROUND:NAFLD includes a spectrum of histological 

phenotypes which can progress to cirrhosis at variable rates. 

While fibrosis assessment with a liver biopsy is the gold standard 

for assessment of disease progression it is limited by 

sampling variability and its invasive nature. There is therefore 

a major unmet need to identify a signature of disease progression 

that does not rely on a liver biopsy. Metabolomics 

provides an unbiased approach to obtain an assessment of 

whole-body metabolic response to disease progression. Previous 

attempts to describe a lipid signature focused on fatty liver 

vs.NASH (Hepatol 2009,50:1827, J Prot Res 2012,11:2521, 

J Lipid Res 2015,56:185) or NASH vs.cirrhosis (J Lipid Res 

2015,56:722). The lipidomic signature of disease progression 

over earlier stages remains unknown. AIM:To characterize 

the changes in the circulating lipidome with disease 

progression from no fibrosis to advanced fibrosis in subjects 

with NAFLD. METHODS:Plasma samples collected at the time 

of liver biopsy were analyzed by UPLC-MS. Specifically, amino 

acids, fatty acyls, bile acids, glycerolipids, glycerophospholipids, 

sterols and sphingolipids were analyzed. RESULTS: 

11controls, 29NAFLD with no fibrosis, 43NAFLD with early 

stage fibrosis (stage1-2) and 22NAFLD with advanced fibrosis 

(stage3-4) were studied. 22metabolites were associated 

with disease progression (Figure). While cholesteryl esters and 

phosphatidylcholines, decreased with the progression [t-test 

Control vs.NAFLD with no fibrosis; early stage or advanced 

fibrosis: PC(O-24:1/0:0) and PC(18:2/20:4):p


0.84), 0.84(0.76-0.92) and 0.88(0.82-0.94), respectively. 

NFS had the highest NPV (98%), while NPVs of AAR, APRI, 

BARD and FIB-4 were 87%, 85%, 94.5% and 94%, respectively. 

56% to 83% of cohort 1 pts were correctly classified as 

having or not AF. In cohort 2 (mean age 46±13, males 65%, 

diabetes 28%, mean BMI 30±5, AF 23.5%), AUROCs of AAR, 

APRI, BARD, FIB-4 and NFS were 0.58(0.50-0.66), 0.55(0.47- 

0.63), 0.64(0.57-0.71), 0.68(0.61-0.75) and 0.66(0.59- 

0.73), respectively. NFS had the highest NPV (84%), while 

NPVs of AAR, APRI, BARD and FIB-4 were 78%, 77.5%, 83% 

and 82%. 64% to 71% of cohort 2 pts were correctly classified 

as having or not AF. In cohort 3 (mean age 49±13, males 

61%, diabetes 38%, mean BMI 31±5, AF 22.7%), AUROCs 

of AAR, APRI, BARD, FIB-4 and NFS were 0.65(0.58-0.71), 

0.73(0.67-0.79), 0.71(0.65-0.76), 0.74(0.68-0.8) and 

0.71(0.65-0.77), respectively. BARD had the higher NPV 

(89%), while NPVs of AAR, APRI, FIB-4 and NFS were 84%, 

81%, 88% and 86%. 61% to 78% of cohort 3 pts were correctly 

classified as having or not AF. Conclusions: There is a 

substantial variability across different cohorts in the diagnostic 

accuracy of non-invasive scoring systems for ruling-out AF in 

NAFLD. Complex scores (BARD, FIB-4, NFS) perform better 

than simple ones (AAR and APRI), but none of them is consistently 

within the acceptable range for clinical-decision making 

in an individual patient. Combination with imaging methods 

should be tested in future studies. 

Disclosures: 



S.A. 

Giulio Marchesini - Advisory Committees or Review Panels: Sanofi-Synthelabo; 

Board Membership: GENFIT, Gilead, Glaxo, Novartis; Grant/Research Support: 

Merck Sharp & Dome; Speaking and Teaching: Novo Nordisk, Merck Sharp & 

Dome, Boerhinger Ingelheim, Eli Lilly, Astra-Zeneca 










The following authors have nothing to disclose: Fabio Nascimbeni, Salvatore 

Petta, Elisabetta Bugianesi, Stefano Bellentani, Christopher P. Day, Pierre 

Bedossa, Claudia P. Oliveira, Raluca Pais 

2165 

DJ-1 labeling index is a novel diagnostic biomarker for 

predicting progression of nonalcoholic steatohepatitis 

Masaaki Takamura 1 , Satoshi Yamagiwa 1 , Yasunobu Matsuda 2 , 

Minoru Nomoto 1 , Toshifumi Wakai 1 , Shuji Terai 1 ; 1 Niigata University 

Graduate School of Medical and Dental Sciences, Niigata, 

Japan; 2 Niigata University Graduate School of Health Sciences, 

Niigata, Japan 

Background & Aims: Nonalcoholic steatohepatitis (NASH) is 

a common cause of chronic liver disease that can progress to 

cirrhosis and hepatocellular carcinoma. We have previously 

reported that hepatic proteins that were dysregulated in a 

murine NASH model functioned in a wide variety of pathways, 

including oxidative stress, glycolysis, the urea cycle, fatty acid 

oxidation, and apoptosis, based on a proteomic approach. 

Among the candidate proteins, we focused on the antioxidant 

protein DJ-1, which was upregulated in the livers of NASH 

model mice. In this study we analyzed the clinical implications 

of DJ-1 in patients with NASH. Methods: Liver biopsies and 

blood samples were obtained from 72 and 38 patients with 

nonalcoholic fatty liver disease (NAFLD) and 8 and 16 control 

subjects, respectively. The hepatic DJ-1 expression and serum 

DJ-1 levels were investigated using immunohistochemistry and 

enzyme-linked immunosorbent assay, respectively. The percentage 

of immunoreactive hepatocyte nuclei in the total number 

of hepatocyte nuclei was defined as DJ-1 labeling index (LI). 

NASH was assessed according to the Brunt classification. 

Results: DJ-1 was localized in the hepatocyte cytoplasm in the 

control and was incrementally translocated to the nucleus in 

NAFLD. A stepwise increase in DJ-1 LI (median [interquartile 

range]) was found from the control group (0 % [0-0.16]) to the 

nonalcoholic fatty liver (NAFL) group (4.34 % [1.50-15.4]) to 

the NASH group (38.5 % [29.6-48.7]; p


viduals with NAFLD and HCC were significantly older (63.5 

± 9.7 vs. 57.4 ± 10.5 years, P=0.0004) and more frequently 

male (67% vs. 38%, P=0.0004). In addition, those with HCC 

were more likely to have esophageal varices (57% vs. 40%, 

P=0.047) and a trend toward a higher frequency of ascites 

(50% vs. 37%, P=0.1). However, the average MELD score was 

lower in those with HCC compared to controls (11.3 ± 3.6 vs. 

13.3 ± 4.6, P=0.05). Individuals with HCC had a trend toward 

a higher prevalence of dyslipidemia (52% vs. 38%, P=0.09) 

and cardiovascular disease (28% vs. 16%, P=0.09) and were 

more likely to use statins (31% vs. 16%, P=0.04). On multivariate 

regression analysis, increasing age (OR 1.09, 95% CI 

1.03-1.15, P=0.002), male gender (OR 4.31, 95% CI 1.69- 

10.97, P=0.002) and the presence of varices (OR 3.93, 95% 

CI 1.43-10.82, P=0.008) were significantly associated with 

HCC. Conclusion: Male gender, increasing age, and the presence 

of varices are associated with HCC in NAFLD cirrhosis. 

Table. Selected characteristics of NAFLD cirrhosis patients with 

and without HCC 

2167 

Disturbance of regulatory T cells, MDSCs and NK cells is 

involved in NASH and mouse model of NASH 

Tatsuki Morosawa, Yasuteru Kondo, Takayuki Kogure, Jun Inoue, 

Yu Nakagome, Osamu Kimura, Yuta Wakui, Tomoaki Iwata, 

Yasuyuki Fujisaka, Teruyuki Umetsu, Tooru Shimosegawa; Tohoku 

University Hospital, Sendai, Japan 

Background and aim: The involvement of lipotoxicity, insulin 

resistance, intestinal bacteria, and disturbance of the immune 

system has been reported to be involved in the pathogenesis 

of non-alcoholic steatohepatitis (NASH). However, the immunological 

dis-regulation in NASH and mouse models of NASH 

has not been clarified yet. The aim of this study is to analyze 

the immunological responses in NASH patients and mouse 

models of NASH. Material and method: Patients: Permission 

for the study was obtained from the ethics committee at our 

university. We analyzed various kinds of lymphocytes in the 

peripheral blood from 35 NASH patients who had been diagnosis 

histologically, eight patients with non-alcoholic fatty liver 

disease, 35 patients with chronic hepatitis C, 20 patients with 

chronic hepatitis B, and 18 healthy subjects. We subjected the 

samples to BD FACS Canto flow cytometry and analyzed; 

(1) myeloid dendritic cells (mDC)1, mDC2, plasmacytoid DC 

(pDC), (2) T cells (regulatory T cells (Tregs), Th1, Th2, Th17), 

(3) Natural Killer (NK) cells, NK-T cells, (4) Myeloid-Derived 

Suppressor Cells (MDSCs). Further, the expression of PD-1, 

PD-L1, NKG2D and CD69 on these immune cells was analyzed 

to detect the level of functional activity. In addition, we 

separated the immune cells from the liver and spleen tissue of 

STAM mice, MCD mice, and C57BL/6 mice (normal diet group 

or high-fat diet group) and conducted an analysis of immune 

functions. Co-cultivation analysis: HepG2 cells with or without 

fatty acid were used for co-cultivation with PBMC to determine 

the effect of fatty hepatocytes on various immune cells. Results: 

We observed a significant increase of Tregs and MDSCs in the 

immune cells of the NASH patients compared with the NAFLD 

patients and healthy subjects (p


(IRR 5.7, p=0.001), but the presence of USS detected hepatic 

steatosis was not. Assessment of diagnostic test accuracy found 

area under the receiver operator curves of 0.7-0.8 for the liver 

fibrosis markers, with optimal cut-offs giving sensitivities of up 

to 90% but positive predictive values of 10% of total body weight lost (OR 

21.67; 95% CI 3.73-125.77; p=0.0006), and change in 

weight measured as mean kilograms lost (mean difference of 

21.0 kg; 95% CI 8.40-33.60; p=0.001), regression of fibrosis 

(OR 18.90; 95% CI 2.10-170.39; p=0.009), reduction in 

ALT (-3.87, 95%CI -3.89-3.85, p=


and 9 reported a change in fibrosis score (D weighted mean 

-0.34). Conclusion: In this meta-analysis, patients with NAFLD 

who underwent bariatric surgery showed significant reduction 

of all-cause mortality, weight loss, fibrosis score and ALT. Further 

studies are needed to assess the longterm benefit of bariatric 

surgery in the treatment of patients with NAFLD. 

Disclosures: 



The following authors have nothing to disclose: Amanda Schneier, Naveen Ganjoo, 

Rachel Pinotti, Malcolm M. Wells 

2171 

Clinical features and risk factors associated with 

Non-alcoholic steatohepatitis (NASH) in Spain. On 

behalf of HEPAmet Registry 

Rocío Gallego-Durán 12 , Rocío Aller 5 , Javier Ampuero 12 , Carmelo 

García-Monzón 1 , Jesus M. Banales 2 , Javier Crespo 3 , Víctor Aguilar-Urbano 

6 , María Luisa García-Torres 7 , José Luís Calleja 4 , Jose 

Luis Olcoz 8 , Javier Salmeron 9 , Martin Prieto 10 , Javier García-Samaniego 

11 , Moisés Diago 7 , Helena Pastor 12 , María J. Pareja 12 , 

Miguel Fernandez-Bermejo 13 , Judith Gomez-Camarero 14 , Raul 

J. Andrade 15 , Pamela Estévez 16 , Conrado M. Fernández-Rodríguez 

17 , Lourdes Grande 18 , Jose María Moreno-Planas 19 , Marta 

Maraver Zamora 20 , Agustin Albillos 21 , Manuel Romero-Gomez 12 ; 

1 Santa Cristina University Hospital, Madrid, Spain; 2 Biodonostia 

Research Institute, San Sebastián, Spain; 3 HUM Valdecillas. 

IDIVAL, Santander, Spain; 4 Puerta de Hierro Hospital, Madrid, 

Spain; 5 Clínico Valladolid University Hospital, Valladolid, Spain; 

6 Costa del Sol Hospital, Málaga, Spain; 7 General de Valencia 

Hospital, Valencia, Spain; 8 León University Hospital, León, Spain; 

9 San Cecilio University Hospital, Granada, Spain; 10 La Fe University 

Hospital, Valencia, Spain; 11 Carlos III Hospital, Madrid, 

Spain; 12 Valme University Hospital and CIBERehd, Sevilla, Spain; 

13 San Pedro de Alcántara Hospital, Cáceres, Spain; 14 Burgos University 

Hospital, Burgos, Spain; 15 Virgen de la Victoria University 

Hospital, Málaga, Spain; 16 Meixoxeiro Hospital, Vigo, Spain; 

17 Fundación Alcorcón University Hospital, Madrid, Spain; 18 Virgen 

del Rocío University Hospital, Sevilla, Spain; 19 Albacete University 

Hospital, Albacete, Spain; 20 Juan Ramón Jiménez University 

Hospital, Huelva, Spain; 21 Ramón y Cajal University Hospital, 

Madrid, Spain 

Aims: To describe the individual profile of NASH Spanish 

patients and to identify risk factors associated with this condition. 

Material and methods: Spanish Association for the 

Study of the Liver (AEEH) HEPAmet Registry is a multicentric 

monitored patient database, including Spanish NAFLD-biopsy 

patients that fulfilled at least 2/4 inclusion criteria (steatosis 

by ultrasound(US); ALT or AST above upper limits of normality(ULN); 

HOMA-IR>4 or metabolic syndrome(MetS) defined 

by ATPIII criteria). Demographic, anthropometric, concomitant 

diseases and medication, US, transient elastography, analytical 

and anatomopathological data were recorded.Univariate 

and multivariate analyses were performed to identify risk factors 

for NASH defined by histology. Results: Seven hundred 

and twenty-six patients were included, 43.8%(318/726) 

men, mean age 51.5+12.9 years old. 50%(370/726) presented 

MetS, 34%(250/726) morbid obese underwent bariatric 

surgery, 73%(528/726) overweight (BMI>25 kg/ 

m 2 ) and 1.2%(9/689) suffered from any cerebrovascular 

event. NASH at liver biopsy was found in 47%(343/726) of 

patients. Fibrosis stage distribution was F0:50%(364/726), 

F1:30%(218/726), F2:9%(65/726), F3:8%(57/726) and 

F4:3%(22/726). Extrahepatic conditions associated with 

NASH vs simple steatosis were T2DM(30.9% vs 21.4%, 

p=0.005) and hyperuricemia(7.8% vs 3.6%, p=0.043). No 

differences were observed between both groups according 

to MetS(55%(182/330) vs 51%(188/371); p=ns) and overweight 

prevalence(76%(250/329) vs 74%(278/376);p=ns). 

In univariate analysis, age,T2DM, arterial hypertension, 

hypercholesterolemia, BMI, AST, ALT, HOMA-IR, transferrin 

saturation index(TSI), platelets and ferritin were significantly 

associated with NASH. In multivariate analysis, HOMA-IR, ALT 

and TSI were confirmed as independent risk factors for NASH. 

Conclusions: The results of this large cohort confirmed insulin 

sensitivity, hepatic inflammation and iron metabolism as key 

factors for NASH. Interestingly, MetS and overweight seems to 

be equally distributed. 

Table 1. Univariate and multivariate analyses in NASH patients 

Disclosures: 


Javier García-Samaniego - Consulting: Bristol-Myers-Squibb, Gilead, Janssen, 

Abbvie 

Moisés Diago - Grant/Research Support: MSD, JANSSEN; Speaking and Teaching: 

BMS, ABBVIE, GILEAD 



S.A. 

The following authors have nothing to disclose: Rocío Gallego-Durán, Rocío Aller, 

Javier Ampuero, Carmelo García-Monzón, Jesus M. Banales, Javier Crespo, Víctor 

Aguilar-Urbano, María Luisa García-Torres, José Luís Calleja, Jose Luis Olcoz, 

Javier Salmeron, Helena Pastor, María J. Pareja, Miguel Fernandez-Bermejo, 

Judith Gomez-Camarero, Raul J. Andrade, Pamela Estévez, Conrado M. Fernández-Rodríguez, 

Lourdes Grande, Jose María Moreno-Planas, Marta Maraver 

Zamora, Agustin Albillos


2172 

WITHDRAWN 

2173 

MHC Class I Related Gene A (MICA) Alleles are Independently 

Associated with Advanced Pathogenic Features 

of Non Alcoholic Fatty Liver Disease (NAFLD) 

Azza Karrar 1 , Mohamad Houry 1 , Irfan Ali 2 , Dinan Abdelatif 2 , 

Pegah Golabi 1 , Mehmet Sayiner 1 , Zahra Younoszai 1 , Lakshmi 

Alaparthi 2 , James N. Cooper 1 , Munkhzul Otgonsuren 1 , Zachary 

D. Goodman 2 , Zobair M. Younossi 2 ; 1 Betty and Guy Beatty Center 

for Integrated Research, Inova Health Systems, Falls Church, VA; 


Hospital, Falls Church, VA 

Background and Aims: MICA is a highly polymorphic gene 

that modulates immune surveillance by binding to its receptor 

on natural killer cells, and its genetic polymorphisms have been 

associated with chronic immune-mediated diseases. NAFLD is 

characterized by fat accumulation in the liver and inflammation 

that can potentially progress to Non Alcoholic Steatohepatitis 

(NASH) and fibrosis. To-date, no data exists describing the role 

of MICA in the pathogenesis of NAFLD. The overall aim is to 

study the association between MICA polymorphism and NASH 

and its histological features. Methodology: DNA from biopsy-proven 

NAFLD patients were genotyped using (PCR-SSO) for 

MICA alleles. Liver biopsies were assessed for the presence of 

NASH, degree of fibrosis and inflammation. Multivariate analysis 

was performed to draw associations between MICA alleles 

and the different variables; p values ≤ 0.05 were considered 

to be significant. Results: The study cohort was comprised of 

134 patients; NAFLD: [NASH (n=32), Age 45.7±10.6 years; 

Male 37.5% and non NASH (n=49), Age 42.5±10.6 years; 

Male 10.2%] and Control: [(n=53), Age 55.1±16 years; Male 

59.6%]. Univariate analysis showed that MICA*007[4(4.9%)] 

and MICA*012[1(1.2%)] were less frequent in NAFLD 

patients, than healthy controls [8(15.1%), p=0.044], [5(9.4%), 

p=0.025] respectively. Furthermore, MICA*011 Odds Ratio 

[OR: 7.14(1.24-41.0), p=0.028] is associated with higher risk 

for NASH. Histologically, MICA*002 was found to be associated 

with lower risk for focal necrosis [OR: 0.34 (0.12-0.92), 

p=0.033], lymphocyte-mediated inflammation [OR: 0.13(0.02- 

1.05), p=0.056] and advanced fibrosis [OR: 0.20(0.04- 

0.92), p=0.039]. In addition, MICA*004 was associated with 

lower risk for portal fibrosis [OR: 0.30(0.09-1.02), p=0.054] 

and portal inflammation [OR: 0.29(0.08-1.05), p=0.059]. On 

the other hand, MICA*017 was associated with higher risk 

for lymphocyte-mediated inflammation [OR: 5.08(1.11-23.2), 

p=0.036]. Multivariate analysis showed that MICA*011 

[OR: 7.38(1.10-49.6), p=0.04] was still independently associated 

with higher risk for NASH. MICA*002 was found to 

be independently associated with lower risk for focal necrosis 

[OR: 0.24(0.08-0.74), p=0.013] and advanced fibrosis [OR: 

0.11(0.02-0.70), p=0.019]. MICA*017 continued to be independently 

associated with higher risk for lymphocyte-mediated 

inflammation [OR: 5.12(1.12-23.5), p=0.035]. Conclusions: 

MICA alleles were found to be independently associated with 

NASH, inflammation and fibrosis. Thus, they may contribute to 

the injury and scarring of liver. Additional research is required 

to investigate the potential role of MICA in suceptability and 

protection to NAFLD. 

Disclosures: 






The following authors have nothing to disclose: Azza Karrar, Mohamad Houry, 

Irfan Ali, Dinan Abdelatif, Pegah Golabi, Mehmet Sayiner, Zahra Younoszai, 

Lakshmi Alaparthi, James N. Cooper, Munkhzul Otgonsuren 

2174 

A 32-gene poor prognosis signature identifies a subset 

of NASH patients at high risk of long-term mortality 

after bariatric surgery 

Nicolas Goossens 1,2 , Won-Min Song 3 , Minoa K. Jung 4 , Philippe 

Morel 4 , Shigeki Nakagawa 5 , Xiaochen Sun 1 , Anu Venkatesh 5 , 

Anna Koh 5 , Jean-Louis Frossard 2 , Laurent Spahr 2 , Francesco 

Negro 2,6 , Scott L. Friedman 1 , Laura Rubbia-Brandt 6 , Emiliano Giostra 

2 , Yujin Hoshida 5 ; 1 Division of Liver Diseases, Department of 

Medicine, Icahn School of Medicine at Mount Sinai, New York, 

NY; 2 Division of Gastroenterology and Hepatology, Geneva University 

Hospital, Geneva, Switzerland; 3 Department of Genetics 

and Genomic Sciences, Icahn Institute of Genomics and Multiscale 

Biology, Icahn School of Medicine at Mount Sinai, New York, NY; 

4 Division of Surgery, Geneva University Hospital, Geneva, Switzerland; 

5 Division of Liver Diseases, Department of Medicine, Liver 

Cancer Program, Tisch Cancer Institute, Icahn School of Medicine 

at Mount Sinai, New York, NY; 6 Division of Pathology, Geneva 

University Hospital, Geneva, Switzerland 

Background & Aims Non-alcoholic steatohepatitis (NASH) is 

associated with increased mortality in subjects undergoing bariatric 

surgery (Goossens et al., EASL 2015). We aimed to 

determine whether bariatric surgery improved survival of obese 

patients with NASH and to identify molecular-based prognostic 

biomarkers in high-risk NASH patients undergoing bariatric 

surgery. Methods We included 492 subjects undergoing Rouxen-Y 

gastric bypass bariatric surgery in a single center between 

January 1997 and December 2004 with a median follow-up 

of up to 17 years. Survival of the entire cohort and of the 

subset of NASH subjects (defined here as steatosis and raised 

ALT for all bariatric surgery and NHANES III subjects) was 

compared to propensity score-matched subjects from the third 

National Health and Nutrition Examination Survey (NHANES 

III). We performed gene expression profiling in formalin-fixed, 

paraffin-embedded hepatic tissue of 47 NASH subjects from 

the bariatric surgery cohort. A prognostic 32-gene signature 

(PMID: 25143343) was evaluated in the NASH subjects and 

gene regulatory networks were identified by using Multiscale 

Embedded Gene Co-expression Network Analysis (MEGENA). 

Results In the bariatric surgery cohort, median body mass index 

(BMI), age, proportion of histologically documented NASH 

were 44kg/m 2 , 41 years, and 12%, respectively. Bariatric 

surgery was associated with reduced mortality compared to 

propensity-score matched NHANES III subjects (HR 0.54, 

p=0.035), however this benefit was not observed in the 

subgroup of NASH patients (HR 0.97, p=0.94). Within the 

47 NASH patients with gene expression analysis (baseline 

BMI 44.6kg/m 2 , 78% with fibrosis, 15% death during follow-up), 

a 32-gene poor prognosis signature prediction was 

associated with increased mortality in multivariable analysis 

(HR 7.7, p=0.045). The 32-gene poor prognosis NASH subjects 

(49%) had increased mortality compared to NHANES 

III NASH subjects (HR 4.4, p=0.005). MEGENA identified 

9 distinct gene regulatory modules associated with TNF-α/ 

NF-κB signaling (p


long-term mortality compared to control obese NASH subjects. 

Gene expression profiling of hepatic tissue in NASH subjects 

revealed the induction of TNF-α /NF-κB and fibrogenic pathways 

as potential targets. 

Disclosures: 













The following authors have nothing to disclose: Nicolas Goossens, Won-Min 

Song, Minoa K. Jung, Philippe Morel, Shigeki Nakagawa, Xiaochen Sun, Anu 

Venkatesh, Anna Koh, Jean-Louis Frossard, Laurent Spahr, Laura Rubbia-Brandt, 

Emiliano Giostra, Yujin Hoshida 

2175 

Naltrexone/Bupropion Extended-Release 32 mg/360 

mg significantly improves liver enzymes in obese/overweight 

individuals with elevated liver enzymes 

Allison Winokur 1 , Amy Halseth 2 , Claire Dybala 1 , Hung Lam 3 , 

Steve Chen 1 , Naga P. Chalasani 4 ; 1 Takeda Pharmaceuticals 

U.S.A., Inc., Deerfield, IL; 2 Orexigen Therapeutics, Inc., La Jolla, 

CA; 3 Takeda Development Center Americas, Inc., Deerfield, IL; 

4 Division of Gastroenterology and Hepatology, Indiana University 


Background: Non-alcoholic fatty liver disease (NAFLD) can be 

a consequence of obesity. Alanine aminotransferase (ALT) is 

often increased in patients with NAFLD and is used as a measure 

of liver dysfunction. NAFLD can improve with weight loss. 

Aim: This posthoc analysis examined the effect of extendedrelease 

naltrexone/bupropion (NB), approved in the US and 

Europe for obesity management, on ALT in non-diabetic, obese 

patients. Methods: Subjects from 3 Phase 3 studies who were 

randomly assigned to receive NB or placebo (PBO), and who 

remained in the trials for 56 weeks were included. NB subjects 

had to lose at least 5% of their body weight at week 16 to 

be included, consistent with prescribing information. Analyses 

by baseline (BL) ALT quartile included change from BL in 

ALT, % of subjects achieving a 25% or 50% reduction in ALT, 

and % subjects achieving Prati criteria (≤19 IU/L for women; 

≤30 IU/L for men) at Week 56. Results: Study population consisted 

of 781 NB subjects (mean age 46 years, BMI 36 kg/ 

m 2 , 85% female, median ALT 22 IU/L) and 663 PBO subjects 

(mean age 46 years, BMI 36 kg/m 2 , 84% female, median ALT 

22 IU/L). NB responders completing 56 weeks of treatment 

experienced significantly greater weight loss vs. PBO (-12.4% 

vs. -2.7%, p


ALT from randomisation to the end of treatment in the metronidazole 

and inulin treatment arm (Group 3) compared to 

the placebo group (group 1), (mean change in ALT -18.6 vs 

-0.5 U/L respectively; p=0.04). The reduction in ALT in group 

2 was not significantly different to the placebo group (mean 

change ALT -2.6 vs 0.5U/L respectively; p=0.5). At the end of 

treatment reduction in BMI, fasting lipids, glycaemia, BP and 

Fibroscan ® CAP scores did not significantly differ between the 

three groups. Conclusions: This is the first clinical trial evidence 

that supplementation with prebiotic inulin following brief metronidazole 

therapy can further reduce liver aminotransferase 

after 4 weeks VLCD therapy in patients with NAFLD. This is a 

potentially viable clinical therapy which requires validation in 

larger clinical studies of longer duration. 

Disclosures: 

The following authors have nothing to disclose: David W. Orr, Rinki Murphy 

2177 

Diagnostic accuracy for non-alcoholic fatty liver disease 

(NAFLD) with controlled attenuation parameter (CAP) 

measured by transient elastography 

Masahiro Kikuchi 1,2 , Rumiko Umeda 2 , Kota Tsuruya 2 , Hirokazu 

Shiozawa 2 , Miho Kikuchi 1 , Masahiko Takahashi 1 , Yoshinori 

Horie 4 , Yasuhiro Nishizaki 3 ; 1 Gastroenterology, National Hospital 

Organization Tokyo Medical Center, Tokyo, Japan; 2 Gastroenterology, 

Tokai University Tokyo Hospital, Tokyo, Japan; 3 Life Care 

Center, Tokai University Tokyo Hospital, Tokyo, Japan; 4 International 

University of Health and Welfare, Research Centre of Clinical 

Medicine, Sanno Medical Center, Tokyo, Japan 

Purpose: We reported that pulse wave velocity (PWV) as arteriosclerosis 

index accelerate and remnant like particles cholesterol 

(RLP-C), triglyceride (TG), and free fatty acid (FFA) which 

are arteriosclerosis promoting factors increase in non-alcoholic 

fatty liver disease (NAFLD). As NAFLD is closely related to 

cardiovascular deaths, a detection of the early stage of NAFLD 

patient is important. In this study, we aimed to investigate the 

diagnostic performance of controlled attenuation parameter 

(CAP) based on FibroScan ® 502 (Echosens,France) in NAFLD 

patients. Method: 341 non-viral hepatitis patients who intake 

alcohol less than 20g ethanol per day were enrolled. The 

existence of the fatty liver was assessed by B-mode ultrasonography, 

and it divided into two groups. (male: NAFLD (+) 

126cases and NAFLD (-) 97cases, female: NAFLD (+) 34cases 

and NAFLD (-) 84cases), and CAP was performed simultaneously. 

Moreover, fatty liver index (FLI) (Bedogni G.2006), 

hepatic steatosis index (HSI) (Lee JH.2010) and lipid accumulation 

product (LAP) (Chinag Jui-Kun.2012) known as liver steatosis 

prediction formulas from various physical measurements 

and multiple blood markers were investigated, and we compared 

the usefulness for detecting NAFLD with ROC analysis. 

Result: CAP value, FLI, HSI, and LAP were significantly high in 

the NAFLD (+) group compared with the (-) group (P< 0.0001). 

In male the area under ROC curve (AUROC) for NAFLD diagnostic 

tool was CAP: 0.890, FLI: 0.779, HSI: 0.811, and LAP: 

0.718 respectively. In female it was CAP: 0.894, FLI: 0.939, 

HSI: 0.911, and LAP: 0.928. The CAP was correlated with 

diagnosis of NAFLD especially in male (cut-off value=257d- 

B/m, sensitivity=87%, specificity=79%). Conclusion: The utility 

of CAP value was verified as accurate diagnosis tool for 

NAFLD. The detection of CAP value would influence the more 

aggressive treatment intervention and the raised motivations of 

therapy for NAFLD patients. This instrument must be contributed 

to not only the detection of NAFLD but also the prevention of 

arteriosclerosis related cardiovascular deaths. 

Disclosures: 

The following authors have nothing to disclose: Masahiro Kikuchi, Rumiko 

Umeda, Kota Tsuruya, Hirokazu Shiozawa, Miho Kikuchi, Masahiko Takahashi, 

Yoshinori Horie, Yasuhiro Nishizaki 

2178 

DACRAs are novel therapeutic candidates for the treatment 

of NAFLD and NASH 

Mette J. Nielsen 1,2 , Sara T. Hjuler 1 , Sofie Gydesen 1 , Morten A. 

Karsdal 1 , Kim Henriksen 1 ; 1 Nordic Bioscience A/S, Herlev, Denmark; 

2 Department of Gastroenterology and Hepatology, Odense 

University Hospital, University of Southern Denmark, Odense, Denmark 

Background and aim: Nonalcoholic fatty liver disease (NAFLD) 

is a significant complication of obesity and in many cases 

it leads to the development of nonalcoholic steatohepatitis 

(NASH). NAFLD is defined by the presence of liver fat accumulation 

exceeding 5% of hepatocytes and is strongly associated 

with the metabolic syndrome. While weight loss is beneficial, 

there are no pharmacological treatments for NAFLD and 

NASH, and novel candidates are intensely sought. DACRAs 

(dual amylin and calcitonin receptor agonists) are novel peptide-based 

drug candidates, which have shown promise in the 

treatment of type 2 diabetes due to potent weight reduction 

and alleviation of insulin resistance. Methods: In this study, we 

used High-Fat Diet (HFD)-induced obese Sprague-Dawley rats 

treated with different doses (0.625-10mg/kg) the DACRA KBP- 

042 for 8 weeks to elucidate the effects of a novel DACRA on 

liver fat accumulation, body weight, insulin/glucagon balance 

and glucose homeostasis. Results: A dose-dependent and sustained 

weight loss was obtained, resulting in a 4.7 % weight 

reduction (20 % vehicle-corrected) in the KBP-042 10 mg/kg 

group with a transient reduction in food intake. The weight 

loss was clearly superior to calorie-matched groups. Corresponding 

with the weight loss, visceral fat depots were significantly 

reduced (perirenal Adipose Tissue (AT): p


2179 

Digoxin provides hepatoprotection through inhibition of 

HIF-1a-NOX4-ROS active pathway 

Xinshou Ouyang 1 , Ji-Yuan Zhang 2 , Dechun Feng 3 , Shi-Ying Cai 1 , 

Irma Garcia-Martinez 1 , Sheng-Na Han 1 , Rafaz Hoque 1 , Yonglin 

Chen 1 , Fu-Sheng Wang 2 , Bin Gao 3 , Natalie J. Torok 4 , Wajahat Z. 

Mehal 1 ; 1 Yale University, New Haven, CT; 2 Liver Failure Therapy 

and Research Center, Beijing 302 Hospital (PLA 302 Hospital), 

Beijing, China; 3 NIAAA, NIH, Bethesda, MD; 4 Gastroenterology 

and Hepatology, UC Davis, Sacramento, CA 

Introduction: Reactive oxygen species (ROS) driven tissue damage 

is common to liver diseases. NADPH oxidase family members 

have shown important contribution to the production of 

hepatic ROS, and this dependent in part to HIF-1a. The cardiac 

glycoside digoxin was identified as potent HIF-1a antagonist 

but its role in liver disease is not well defined. Aim: To assess 

whether digoxin has therapeutic effects in Non Alcoholic Fatty 

Liver Disease (NASH) and alcoholic liver disease (ALD) in 

mice, and investigate the molecular mechanisms in both mouse 

and human cell cultures. Methods: C57BL/6J male mice were 

placed on a 45% high fat diet (HFD) for 11weeks with and without 

digoxin (ip 1, 0.2 and 0.05 mg/kg twice a week). Digoxin 

1mg/kg ip daily in mice results in the therapeutic serum levels 

achieved in humans (0.5-2 ng/ml). Plasma ALT, liver histology, 

neutrophil staining, leukocytes profiling, mitochondrial reactive 

oxygen species (ROS) generation, and gene transcriptome 

microarrays were analyzed. The chronic plus binge model of 

ALD was performed. The mechanism of digoxin on inhibition 

of HIF-1a mediated NADPH oxidase 4 (NOX4) transcription 

and ROS production was tested by RT-PCR, reporter luciferase 

and ChIP-PCR assay. Results: Digoxin dose-dependently 

reduced histological injury, neutrophilic infiltrate and serum 

ALT values in both co-treatment (starting digoxin same time 

with HFD, ALT, 417 +/- 398 U/L in HFD vs 91 +/- 73 U/L 

in HFD+DIG, P< 0.001) and post-treatment (starting digoxin 

after 4 weeks HFD, neutrophil 24.6% in HFD vs 14.3% in 

HFD+DIG; monocytes 31.6% in HFD vs 19.1% in HFD+DIG; 

ALT, 400 +/- 130 U/L in HFD vs 80 +/- 17 U/L in HFD+DIG, 

P< 0.001) without a reduction in food intake. A low dose of 

digoxin (0.05 mg/kg) also shows significant protective effects 

again injury and inflammation in both NASH and ALD models. 

Further microarray analysis in HFD liver tissues revealed 

that digoxin down-regulated ROS metabolism and antioxidant 

pathway including NOX4. In vitro data showed that digoxin 

dose-dependently inhibited mitochondrial ROS production 

under TLR and hydrogen peroxide stimulation in multiple mouse 

and human cell cultures. Digoxin increases VHL protein levels, 

resulting in a corresponding reduction HIF-1a protein levels. In 

addition digoxin blocks HIF-1a mediated NOX4 gene expression, 

promoter activity and NOX4 mediated ROS. Digoxin 

also reduces HIF-1a binding to the NOX4 promoter region. 

Conclusions: Digoxin has strong effects in reducing liver steatosis 

and inflammation in experimental models of NASH and 

ALD via a HIF-1a-NOX4-ROS pathway. Low dose digoxin may 

provide significant therapeutic value in the treatment of NASH 

and ALD patients. 

Disclosures: 

The following authors have nothing to disclose: Xinshou Ouyang, Ji-Yuan Zhang, 

Dechun Feng, Shi-Ying Cai, Irma Garcia-Martinez, Sheng-Na Han, Rafaz Hoque, 

Yonglin Chen, Fu-Sheng Wang, Bin Gao, Natalie J. Torok, Wajahat Z. Mehal 

2180 

Can FibroMeter VCTE be useful for detecting hepatic 

fibrosis in patients with non-alcoholic fatty liver disease? 

Elif Dincses 3 , Yusuf Yilmaz 1,2 ; 1 Department of Gastroenterology, 

Marmara University, School of Medicine, Istanbul, Turkey; 2 Liver 

Research Unit, Institute of Gastroenterology, Marmara University, 

Istanbul, Turkey; 3 Department of Internal Medicine, Marmara University, 

Istanbul, Turkey 

Hepatic fibrosis plays a paramount role in determining the 

prognosis of patients with non-alcoholic fatty liver disease 

(NAFLD). Originally developed for the non-invasive detection 

of fibrosis in patients with chronic viral hepatitis, the FibroMeter 

VTCE is a diagnostic tool comprising both biochemical 

markers and transient elastography (TE). We conducted this 

pilot study to investigate the diagnostic performance of the 

FibroMeter VTCE tool for diagnosing fibrosis in patients with 

biopsy-proven NAFLD and compare its diagnostic accuracy 

with those of the NAFLD fibrosis score (NFSA) and TE alone. 

We determined FibroMeter VTCE, NFSA, and TE in 52 

NAFLD patients. The results of liver biopsies were considered 

as the gold standard. Areas under the ROC curve (AUROCs) 

were used to express the diagnostic accuracy of each test. 

We detected significant (F≥2) and severe (F≥3) fibrosis in 20 

(38%) and 10 (19%) patients, respectively. The sensitivity of 

FibroMeter VTCE, NFSA, and TE for detecting significant 

fibrosis was 70%, 65%, and 75%, respectively, whereas specificity 

was 88%, 81%, and 78%. The sensitivity of FibroMeter 

VTCE, NFSA, and TE for diagnosing severe fibrosis was 

90%, 90%, and 100%, respectively, whereas specificity was 

93%, 78%, and 76%. ROC analysis showed that FibroMeter 

VTCE had a significantly larger AUROC (0.968) compared 

to both NFSA (0.833, P < 0.001) and TE (0.922, P < 0.05) for 

the detection of severe fibrosis (Table). To the best of our knowledge, 

this is the first study showing that FibroMeter VTCE – a 

tool originally developed for patients with viral hepatitis – can 

be superior to both NFSA and TE for the detection of severe 

hepatic fibrosis in patients with NAFLD. Financial disclosures: 

The authors declare no conficts of interest. 

AUROCs and sensitivities/specificities of FibroMeter VTCE, 

NFSA, and TE for the detection of severe fibrosis (F≥3) in patients 

with biopsy-proven non-alcoholic fatty liver disease 

Abbreviations: AUROC = area under the receiver operating 

characteristic curve; NFSA = NAFLD fibrosis score; TE = transient 

elastography. 

Disclosures: 

The following authors have nothing to disclose: Elif Dincses, Yusuf Yilmaz


2181 

Controlled Attenuation parameter (CAP) for the diagnosis 

of steatosis in NonAlcoholic Fatty Liver Disease. 

Jean-Baptiste Hiriart 1 , Grace Wong 2 , Julien Vergniol 1 , Henry Lik- 

Yuen Chan 2 , Brigitte Le Bail 3 , Anthony W. Chan 4 , Faiza Chermak 

1 , Juliette Foucher 1 , Wassil Merrouche 1 , Angel ML Chim 2 , 

Victor de Ledinghen 1 , Vincent W. Wong 2 ; 1 Hepatology, Hôpital 

Haut-Lévêque, Pessac, France; 2 Medicine and Therapeutics, Prince 

of Wales Hospital, Hong Kong, China; 3 Pathology, Hôpital Pellegrin, 

Bordeaux, France; 4 Anatomical and Cellular Pathology, 

Prince of Wales Hospital, Hong Kong, China 

Background & Aims: Controlled attenuation parameter (CAP) 

evaluated with transient elastography (FibroScan) is a recent 

method for non-invasive assessment of steatosis. Its usefulness 

in NonAlcoholic Fatty Liver Disease (NAFLD) is unknown. We 

prospectively investigated the performance of CAP for the 

diagnosis of steatosis in NAFLD. Methods: From April 2009 to 

June 2014, all CAP examinations performed in adult NAFLD 

patients with a liver biopsy performed within one week of CAP 

measurement were included. Liver biopsies were assessed for 

necro-inflammatory activity and fibrosis stage, NAFLD activity 

score (NAS), and steatosis graded as follows: S0, steatosis 

≤5% of hepatocytes; S1, 6-33%; S2, 34-66%; S3, >66%. 

The Fatty Liver Index (FLI) was calculated on the day of liver 

biopsy. Receiver-operating characteristics (ROC) curves were 

constructed to assess the overall accuracy of CAP. Results: 

A total of 261 consecutive patients (59% males, mean age 

56 years) from two ethnic groups with CAP measurement and 

satisfactory liver biopsy specimens were included. Mean BMI 

was 30,2 ± 5,1 kg/m 2 , 59% of patients had diabetes and 

65,9% had metabolic syndrome (according to the International 

Diabetes Federation criteria). Mean length of liver biopsy was 

25.1 ± 8.6 mm. 46% of patients had NAS score ≥ 5. Fibrosis 

stages were distributed as follows: 37,9% F0F1, 21,5% F2, 

21,8% F3 and 18,8% F4. All patients had steatosis > 5%, 

29,9% were graded S1, 38,3% S2 and 31,8% S3. The area 

under the ROC curve of CAP for steatosis ≥ S1, ≥ S2 and S3 

was 0.85, 0.80, and 0.66 respectively. At a cutoff value of 

310 dB/m, the sensitivity, specificity, positive and negative 

predictive values for ≥ S2 steatosis were 79%, 71%, 86% and 

71%, respectively. CAP had significantly better performance 

than FLI for the diagnosis of S2S3 steatosis (0,8 [0,73 – 0,86] 

vs 0,66 [0,59 – 0,74]; p < 0,001). Discordance of at least 

one grade between CAP and steatosis was observed in 81 

(31%) patients. By multivariate analysis, only steatosis S2S3 

was associated with no discordance. Only 15% of patients 

with NAS ≥ 5 had discordance between CAP and steatosis 

grade. By multivariate analysis, only BMI ≥ 30 kg/m 2 was 

significantly associated with CAP > 310 dB/m. Conclusion: 

CAP provides an immediate assessment of steatosis simultaneously 

with liver stiffness measurement. The strong association 

of CAP with steaosis, especially in patients with nonalcoholic 

steatohepatitis, and with elevated BMI could be of interest for 

the diagnosis and follow-up of NAFLD patients. 

Disclosures: 

Grace Wong - Speaking and Teaching: Echosens, Echosens, Echosens, Echosens 




Juliette Foucher - Board Membership: BMS, Gilead, Abbvie; Speaking and 

Teaching: BMS, Gilead, Janssen, AbbVie 





Echosens 

The following authors have nothing to disclose: Jean-Baptiste Hiriart, Julien 

Vergniol, Brigitte Le Bail, Anthony W. Chan, Faiza Chermak, Wassil Merrouche, 

Angel ML Chim 

2182 

Obesity fibrosis score: A new model to predict 

advanced fibrosis in morbidly obese patients with nonalcoholic 

fatty liver disease (NAFLD) 

Tavankit Singh, Srinivasan Dasarathy, Rocio Lopez, Arthur J. 

McCullough; Cleveland Clinic Foundation, Cleveland, OH 

Introduction: The prevalence of NAFLD is high in obese 

patients. Current diagnostic tests include a liver biopsy that 

is invasive and fibroscan whose cost and lack of validation in 

the morbidly obese population makes it necessary to evaluate 

subjects by other non-invasive measures. Although a number 

of prediction models (APRI, BARD, FIB-4 and NAFLD fibrosis 

scores) have been developed, they have not been validated 

in morbidly obese patients. We designed this study to develop 

a predictive model for advanced fibrosis in morbidly obese 

patients and to compare it with the other known models. Methods: 

Patients with morbid obesity (BMI >40 kg/m 2 or >35 

kg/m 2 with diabetes) and NAFLD proven by biopsy between 

2005-2014 were included and randomly split into two groups 

for model building (training cohort) and model validation (validation 

cohort) in a 2:1 ratio. To build the prediction model, 

multivariable logistic regression analysis was performed. Internal 

model validation was measured by area under receiver 

operating curve (AUROC). Results: 612 patients (408 and 

204 patients in training and validation cohorts respectively) 

were included in the study. Both cohorts had similar clinical 

profiles, laboratory values, patient demographics. 25% 

patients had advanced fibrosis. After analysis, the model to 

predict advanced fibrosis was defined as follows: z = -27303 

+(1.2671 if diabetic) +(0.0162XALP) - (0.0072X platelets) 

+(0.0165XAST) +0.6359 if AST/ALT ≥0.8 This value was then 

converted into a probability distribution with a value between 

0 and 100. AUROC was 0.756 and 0.808 in training and 

validation cohorts respectively. Scores of 44.2 and 14.6 were 

used to include or exclude the presence of advanced fibrosis 

and provided a sensitivity and specificity of 90% and 44% 

for the lower cut-off and 26% and 90% for the upper cut-off, 

respectively. This model was superior to APRI, FIB-4, BARD, 

NAFLD fibrosis scores in both the cohorts. Compared to FIB- 

4, the difference was not significantly different. However, the 

FIB-4 score could only identify 8/102 patients with advanced 

fibrosis thus making it less useful. Conclusion: In the obese 

population, the obesity fibrosis score is superior to the currently 

existing prediction models to detect advanced fibrosis and will 

permit careful selection of patients for further diagnostic evaluation. 

Prediction Scores for Advanced Fibrosis:ROC Analysis 

Disclosures: 

The following authors have nothing to disclose: Tavankit Singh, Srinivasan 

Dasarathy, Rocio Lopez, Arthur J. McCullough


2183 

Exercise reduces liver fat, visceral fat and circulating 

triglycerides, but not inflammatory markers in non-alcoholic 


David Houghton, Christian Thoma, Kate Hallsworth, Kieren G. 

Hollingsworth, Christopher P. Day, Quentin M. Anstee, Michael I. 

Trenell; Institute of Cellular Medicine, Newcastle University, Newcastle 

Upon Tyne, United Kingdom 

Background: Non-alcoholic steatohepatitis (NASH) represents 

steatosis and ballooning degeneration, inflammation and fibrosis. 

Current treatments for NASH are limited, with no studies 

to date reporting the effects of exercise in people with NASH. 

Studies in NAFLD show promising effects on liver lipid but the 

effect on inflammation and visceral fat, key mediators of fibrosis, 

are not known. Aims: Determine the effect of a 12-week 

exercise intervention on liver lipid and circulatory inflammation 

in people with NASH. Methods: 24 participants (mean age 

52 ± 14 years, BMI 33 ± 6) with histologically characterised 

NASH (NAFLD Activity Score ≥ 5) received either: exercise 

(n = 12) or continue standard care (n = 12) over 12 weeks 

and maintained baseline weight. Participants were not undergoing 

insulin sensitising treatment, dietary change or regular 

activity. Subjects with heart/kidney disease or in vivo ferrous 

material were excluded. Liver lipid content, subcutaneous and 

visceral adiposity were assessed using magnetic resonance 

techniques, inflammation, fibrosis markers, insulin sensitivity 

were assessed at baseline and at 12 weeks. Results: Resistance 

exercise produced a significant reduction in liver lipid content 

(-13 ± 24 vs. 6 ± 16%, P0.05), HOMA 

IR (2.3 ± 1.4 to 1.9 ± 0.8 vs. 1.9 ± 1.1 to 1.7 ± 1.1, P>0.05) 

or 2hour glucose levels or liver enzymes (ALT: 53 ± 25U/L to 

52 ± 18U/L vs. 81 ± 59U/L to 71 ± 52U/L; AST: 41 ± 14U/L 

to 45 ± 12U/L vs. 59 ± 41U/L to 58 ± 45U/L, P>0.05). Conclusion: 

This is the first study reporting the effects of exercise on 

liver lipid, body composition and circulating inflammation in 

patients with histologically defined NASH. Exercise produces 

a significant reduction in liver lipid, visceral fat and plasma triglycerides 

but no effect on body weight, abdominal adiposity, 

or circulatory markers of inflammation. These results suggest 

that exercise alone may be insufficient to target the mediators 

of NASH and warrants further exploration. 

Disclosures: 

Quentin M. Anstee - Advisory Committees or Review Panels: Genfit, Intercept, 

Raptor; Grant/Research Support: GSK; Speaking and Teaching: Abbott Laboratories 

The following authors have nothing to disclose: David Houghton, Christian 

Thoma, Kate Hallsworth, Kieren G. Hollingsworth, Christopher P. Day, Michael 

I. Trenell 

2184 

The link between intestinal microbiota, bile acids and 

non-alcoholic fatty liver disease 

Alice Y. Wang 1 , Marialena Mouzaki 2,3 , Bianca M. Arendt 4,5 , 

Robert Bandsma 2,3 , Scott Fung 4,5 , Sandra E. Fischer 4,6 , Johane 

P. Allard 4,5 ; 1 Physiology and Experimental Medicine Program, 

Research Institute, The Hospital for Sick Children, Toronto, ON, 

Canada; 2 Division of Gastroenterology, Hepatology and Nutrition, 

The Hospital for Sick Children, Toronto, ON, Canada; 3 University 

of Toronto, Toronto, ON, Canada; 4 Toronto General Hospital, 

University Health Network, Toronto, ON, Canada; 5 Department of 

Medicine, University of Toronto, Toronto, ON, Canada; 6 Department 

of Laboratory Medicine and Pathobiology, University of 

Toronto, Toronto, ON, Canada 

Purpose: Intestinal microbiota (IM) may be implicated in the 

pathogenesis of non-alcoholic fatty liver disease (NAFLD) 

through their effects on bile acid (BA) homeostasis. To better 

elucidate the associations, this study characterizes the fecal 

BA profiles of patients with NAFLD and examines the relationship 

between IM and fecal BA composition. Methods: This was 

a prospective, cross-sectional study. The fecal BA profiles of 

adults with biopsy-confirmed NAFLD (non-alcoholic fatty liver: 

NAFL and non-alcoholic steatohepatitis: NASH) and healthy 

controls (HC) were examined. In a subset of subjects, IM composition 

was assessed. Clinical and laboratory data, stool and 

blood samples and 7-day food records were collected prior 

to liver biopsy. Fecal BAs were quantified by liquid chromatography-tandem 

mass spectrometry . Quantitative real-time 

polymerase chain reaction was used to measure total bacterial 

counts, Bacteroides/Prevotella (Bacteroidetes), C. leptum, 

C. coccoides, Bifidobacteria, Escherichia coli and Archaea 

in stool. Statistical analyses were conducted using SPSS v.22 

(IBM Corp 2013). Results: This study included 53 participants 

(25 HC, 11 NAFL, 17 NASH), 56.5% male, with a mean 

(±SD) age of 41.0 ± 10.9 years and a median (IQR) BMI of 

28.4 (23.4-32.4) kg/m 2 . Total fecal BA levels were higher 

in patients with NAFLD (NASH and NAFL combined) compared 

to HC (log values: 3.65 and 3.50 pmol/mg, respectively; 

p=0.021). The ratio of conjugated to unconjugated BA 

was not different between the groups. Patients with NAFLD 

had a higher ratio of primary to secondary BA compared to 

HC. Levels of unconjugated cholic acid (CA) were higher in 

both NASH and NAFL compared to HC (p=0.005, p=0.026, 

respectively), whereas chenodeoxycholic acid (CDCA) was 

higher in NASH vs. HC (p=0.011). The primary conjugated BA 

were not different between the groups; however, patients with 

NASH had higher levels of conjugated lithocholic acid (LCA) 

than patients with NAFL (p=0.035). In a subset of 29 patients 

(18 HC, 11 NAFLD) with IM data, patients with NAFLD had 

decreased levels of Bacteroidetes and C. leptum (p=0.041; 

p=0. 006), which remained significant after adjusting for BMI 

and percent of fat intake. Bacteroidetes levels were inversely 

correlated with glycoLCA (r=-0.550, p=0.002) and tauroLCA 

(r=-0.408, p=0.048). C. leptum levels were positively correlated 

with fecal LCA (r=0.526, p = 0.003) and inversely with 

CA (r=-0.669, p


2185 

Serum marker of inflammasome activity correlates with 

liver injury in nonalcoholic fatty liver disease and is 

influenced by genetic polymorphisms. 

Leon A. Adams 1,2 , Alexander Wree 3 , Phillip Melton 4 , Gary P. 

Jeffrey 1,2 , Helena Ching 2,1 , Bastiaan de Boer 5 , John K. Olynyk 6 , 

Oyekoya T. Ayonrinde 6 , Trevor A. Mori 1 , Lawrence Beilin 1 , Patricia 

Price 7 , Craig E. Pennell 9 , Mohammed Eslam 8 , Jacob George 8 , 

Ariel E. Feldstein 3 ; 1 Medicine and Pharmacology, University of 

Western Australia, Nedlands, WA, Australia; 2 Hepatology, Sir 

Charles Gairdner Hospital, Nedlands, WA, Australia; 3 Pediatrics, 

University of California, San Diego, San Diego, CA; 4 Center of 

Genetic Origins of Health and Disease, The University of Western 

Australia, Nedlands, WA, Australia; 5 Anatomical Pathology, 

PathWest, Nedlands, WA, Australia; 6 Gastroenterology, Fiona 

Stanley Hospital, Perth, WA, Australia; 7 School of Biomedical Sciences, 

Curtin University, Perth, WA, Australia; 8 Medicine, University 

of Sydney, Sydney, NSW, Australia; 9 Womens and Childrens 

Health, The University of Western Australia, Perth, WA, Australia 

Background and Aims: The NLRP3 inflammasome has been 

implicated in the pathogenesis of liver injury in nonalcoholic 

fatty liver disease (NAFLD). In humans, mutations of Nlrp3 

have functional consequences and result in auto-inflammatory 

syndromes. We examined whether single nucleotide polymorphisms 

(SNP’s) of the NLRP3 and CARD8 genes of the NLRP3 

inflammasome influenced serum inflammatory markers and 

fatty liver in a general population based cohort. Secondly, we 

examined serum caspase-1 as a novel marker of liver injury 

and determined the association between inflammasome SNP’s, 

serum caspase-1 and liver injury. Methods: Inflammatory markers 

(CRP, IP-10, TNFR1, TNFR2, IL18), liver enzymes and 

ultrasound diagnosed fatty liver were assessed in a population-based 

cohort (n=1046) of 17-year old adolescents. Serum 

caspase-1 activity and protein levels were determined in a 

second cohort (n=154) and sub-group (n=20) of subjects with 

biopsy proven NAFLD by flourometric assay (Abcam, Cambridge, 

MA) and western blot respectively. Liver histology was 

evaluated according to the NASH CRN scoring system. Eleven 

NLRP3 and CARD8 SNPs were genotyped in both cohorts and 

assessed for phenotypic associations. Results: In cohort 1, the 

rs2043211 SNP in the CARD8 gene was associated with serum 

levels of IP10 (p=0.002), IL18 (p=0.009), sTNFR1 (p=0.04), 

sTNFR2 (p=0.04), CRP (p=0.056) and a diagnosis of fatty 

liver (adj. odds ratio 1.65, 95% CI 1.09-2.48, dominant allelic 

model). Serum IP10 and TNFR2 levels but not SNP genotypes 

were positively correlated with hepatic aminotransaminase 

levels. In cohort 2, serum caspase-1 activity was significantly 

increased in the presence of hepatocellular ballooning, inflammation 

and fibrosis (p


2187 

Liver Fat and Inflammation Are Associated with Circulating 

PCSK9 Levels 

Paola Dongiovanni 1 , Massimiliano Ruscica 2 , Nicola Ferri 2 , Claudia 

Lanti 2 , Raffaela Rametta 1 , Anna Ludovica Fracanzani 1 , Paolo 

Magni 2 , Silvia Fargion 1 , Luca Valenti 1 ; 1 Internal Medicine, University 

of Milano, Fondazione IRCCS Ca Granda, Milano, Italy; 

2 University of Milan, Milan, Italy 

Background and aims: Hepatocellular fat accumulation defines 

nonalcoholic fatty liver disease (NAFLD), also known as hepatic 

steatosis. Lipids accumulation derives from dietary fatty acids, 

increased peripheral lipolysis due to adipose tissue insulin resistance 

and elevated hepatic de novo lipogenesis due to hyperinsulinemia. 

Altered secretion of very low-density lipoproteins 

(VLDL) and free cholesterol accumulation also mediate hepatocellular 

damage. However, NAFLD also confers increased risk 

of cardiovascular events independently of classic risk factors. 

Proprotein Convertase Subtilisin/Kexin type 9 (PCSK9) is an 

important player in lipoprotein metabolism. Cleaved PCSK9 is 

secreted by hepatocytes with VLDL and circulate with lipoproteins. 

Soluble PCSK9 inhibits the uptake of low-density lipoproteins 

(LDL) by targeting LDL receptor (LDL-R) to degradation, and 

loss-of-function variants are associated with reduced circulating 

cholesterol and protection from cardiovascular disease. However, 

it is not known whether hepatic fat content and inflammation 

modify PCSK9 levels. Aim was to evaluate whether hepatic 

fat content affect circulating PCSK9 levels in individuals, who 

underwent liver biopsy for suspected nonalcoholic steatohepatitis 

(NASH). Methods: We considered 203 patients of Italian 

ancestry who underwent biopsy for suspected NASH (n=126 

for persistently elevated serum ALT, n=77 for severe obesity). 

Serum PCSK9 levels were measured using a commercial ELISA 

kit (R&D Systems, Minneapolis, MN, USA). Results: In NAFLD 

patients, at univariate analysis PCSK9 levels were significantly 

associated with steatosis severity (estimate +0.14±0.05 

log ng/mL; p=0.006), inflammation (estimate +0.15±0.06; 

p=0.01), and fibrosis (estimate +0.13±0.04; p=0.001). At 

multivariate analysis, PCSK9 serum levels were significantly 

associated with steatosis grade (estimate +0.13±0.06 log ng/ 

mL; p=0.03), age (estimate +0.008±0.004; p=0.03), BMI 

(estimate -0.016±0.006; p=0.01) independently of sex, and 

liver disease severity. In a subset of 30 obese patients, there 

was no significant association between circulating PCSK9 concentration 

and hepatic mRNA levels. Conclusions: Modulation 

of serum PCSK9 by post-transcriptional mechanisms might be 

involved in mediating the susceptibility to dyslipidemia and atherosclerosis 

progression in patients with NAFLD, with potential 

therapeutic implications. 

Disclosures: 

The following authors have nothing to disclose: Paola Dongiovanni, Massimiliano 

Ruscica, Nicola Ferri, Claudia Lanti, Raffaela Rametta, Anna Ludovica Fracanzani, 

Paolo Magni, Silvia Fargion, Luca Valenti 

2188 

Increasing severity of NAFLD is associated with gut dysbiosis 

and modification of the metabolic function of the 

gut microbiota 

Jerome Boursier 1,3 , Olaf Mueller 2 , Matthieu Barret 4 , Mariana V. 

Machado 5 , Lionel Fizanne 3 , Cynthia D. Guy 6 , John F. Rawls 2 , Lawrence 

David 2 , Gilles Hunault 3 , Frederic Oberti 1,3 , Paul Cales 1,3 , 

Anna Mae Diehl 5 ; 1 Hepato-Gastroenterology Department, University 

Hospital, Angers, France; 2 Center for Genomics of Microbial 

Systems, Duke University, Durham, NC; 3 Laboratoire HIFIH 

EA3859, University, Angers, France; 4 UMR1345, INRA, Beaucouzé, 

France; 5 Division of Gastroenterology, Duke University 

Medical Center, Durham, NC; 6 Department of Pathology, Duke 

University, Durham, NC 

Background: Several animal studies have suggested the role of 

gut microbiota in Non-Alcoholic Fatty Liver Disease (NAFLD). 

However, data about gut dysbiosis in human NAFLD remains 

scarce in the literature, especially studies including the whole 

spectrum of NAFLD lesions. We aimed to evaluate the association 

between gut dysbiosis and the severity of NAFLD, 

i.e. Non-Alcoholic Steatohepatitis (NASH) and fibrosis, in a 

well-characterized cohort of adult NAFLD. Methods: 57 patients 

with biopsy-proven NAFLD were enrolled. The taxonomic composition 

of gut microbiota was determined using 16S ribosomal 

RNA sequencing of stool samples. Results: 30 patients 

had F0/1 fibrosis stage at liver biopsy (10 with NASH), and 

27 patients had significant F ≥2 fibrosis (25 with NASH). 

Bacteroides were significantly increased in NASH and F ≥2 

patients, whereas Prevotella were decreased. Ruminococcus 

were significantly higher in F ≥2 patients. Multivariate analysis 

including age, sex, metabolic syndrome, serum transaminases, 

gammaGT, ultrasensitive CRP, and gut microbiota data showed 

that Bacteroides abundance was independently associated 

with NASH and Ruminococcus with F≥2 fibrosis. Stratification 

according to the abundance of these 2 bacteria generated 

3 patient subgroups with increasing severity of NAFLD 

lesions (low NASH/low fibrosis, high NASH/low fibrosis, high 

NASH/ high fibrosis). Patients with a metabolic syndrome had 

more severe liver lesions and, in those patients, NAFLD severity 

increased according to the 3 gut microbiota subgroups. Based 

on predicted metagenomics profile, the severity of NAFLD was 

associated with significant microbiota enrichment in KEGG 

pathways involved in carbohydrate metabolism (starch and 

sucrose metabolism, pentose phosphate pathway, pentose and 

glucoronate interconversions, pyruvate metabolism, glyoxylate 

and dicarboxylate metabolism), lipid metabolism (lipid biosynthesis 

proteins, sphingolipid metabolism), and amino acid 

metabolism (arginine and proline metabolism, cyanoamino 

acid metabolism). Conclusion: NAFLD severity associates with 

gut dysbiosis and altered metabolic functions of the gut microbiota. 

Bacteroides is an independent predictor for NASH and 

Ruminococcus for fibrosis. Thus, gut microbiota analysis adds 

information to classical predictors of NAFLD severity and suggests 

novel metabolic targets for pre/pro-biotic therapies. 

Disclosures: 

Frederic Oberti - Speaking and Teaching: LFB, gore 


The following authors have nothing to disclose: Jerome Boursier, Olaf Mueller, 

Matthieu Barret, Mariana V. Machado, Lionel Fizanne, Cynthia D. Guy, John F. 

Rawls, Lawrence David, Gilles Hunault, Anna Mae Diehl


2189 

Prevalence of non-alcoholic fatty liver disease (NAFLD) 

and its subtypes obtained by liver biopsy in patients 

with type 2 diabetes not selected by ultrasound and/or 

aminotransferase levels 

Lilian M. Silva 2 , Fatima F. Figueiredo 2,3 , Frederico F. Campos 4 , 

Marcio Q. Miguez 2 , Valéria P. Lanzoni 5 , Marilia B. Gomes 6 , Carlos 

Terra 2 , Hugo Perazzo 1 , Renata M. Perez 2,7 ; 1 National Institute 

of Infectious Diseases Evandro Chagas, Rio de Janeiro, Brazil; 

2 Liver Unit, University of the State of Rio de Janeiro, Rio de Janeiro, 

Brazil; 3 Internal Medicine, Federal University of Rio de Janeiro, 

Rio de Janeiro, Brazil; 4 Pathology, University of the State of Rio 

de Janeiro, Rio de Janeiro, Brazil; 5 Pathology, Federal University 

of São Paulo, São Paulo, Brazil; 6 Diabetology, University of the 

State of Rio de Janeiro, Rio deJaneiro, Brazil; 7 D’Or Institute for 

Research and Education, Rio de Janeiro, Brazil 

Background: Non-alcoholic fatty liver disease (NAFLD) and 

its severe form, steatohepatitis (NASH), have been described 

as highly prevalent in patients with metabolic diseases. The 

aims were: (i) to estimate the prevalence of NAFLD and its 

spectrum by liver biopsy in type 2 diabetes patients; (ii) to 

quantify steatosis and fibrosis; (iii) to identify variables predictive 

of NASH and fibrosis. Patients and methods: Patients with 

type 2 diabetes were submitted to percutaneous liver biopsy 

regardless of alanine aminotransferase (ALT) levels or abdominal 

ultrasound results. The exclusion criteria were presence of 

other chronic liver disease or serious systemic diseases; coagulation 

disturbances; morbid obesity; alcohol abuse; use of 

hepatotoxic drugs; pregnancy; or refusal to participate. Slides 

of liver biopsy were evaluated by two independent pathologists 

using the NASH Activity Score. Factors associated with NASH 

and significant fibrosis (stage F≥2) were defined by logistic 

regression analysis. Results: 85 patients [82% female, mean 

age=54 yrs, mean BMI=31 Kg/m 2 , 61% with normal ALT, 

53% treated by insulin] were included. Median (range) liver 

specimen length was 30 (25-75) mm. Prevalence of NAFLD in 

type 2 diabetes was 92% [n=78/85], 54% (n=42) of whom 

with simple steatosis and 46% (n=36) with NASH. Severe steatosis 

(>33% of hepatocytes) was more prevalent in patients 

with NASH compared to those with simple steatosis (65% vs 

7%; p


Rajarshi Banerjee - Board Membership: Perspectum Diagnostics; Employment: 

Perspectum Diagnostics; Grant/Research Support: Perspectum Diagnostics; Patent 

Held/Filed: Perspectum Diagnostics Ltd, University of Oxford; Stock Shareholder: 

Perspectum Diagnostics 

Elizabeth M. Tunnicliffe - Patent Held/Filed: Perspectum Diagnostics; Stock Shareholder: 

Perspectum Diagnostics 

Stefan Neubauer - Board Membership: Perspectum Diagnostics; Patent Held/ 

Filed: University of Oxford 

The following authors have nothing to disclose: Jane Collier, Lai Mun Wang, 

Fleming A. Kenneth, Jeremy F. Cobbold, Eleanor Barnes 

2191 

Non-invasive hepatic fibrosis markers and cardiometabolic 

risk among adults in the Framingham Heart Study 

Michelle T. Long 1,2 , Alison Pedley 2 , Joseph M. Massaro 3,2 , Udo 

Hoffmann 4 , Caroline S. Fox 2,5 ; 1 Gastroenterology, Boston Medical 

Center, Boston, MA; 2 National Heart, Lung, and Blood Institute’s 

Framingham Heart Study, Framingham, MA; 3 Biostatistics, Boston 

University School of Public Health, Boston, MA; 4 Radiology, 

Massachusetts General Hospital, Boston, MA; 5 Endocrinology, 

Brigham and Women’s Hospital, Boston, MA 

Background & Aims: Non-alcoholic fatty liver disease (NAFLD) 

is associated with an increased risk of cardiovascular related 

death, particularly in those with a high predicted risk for hepatic 

fibrosis. We determined (a) the prevalence of predicted hepatic 

fibrosis based on various non-invasive fibrosis markers and (b) 

the association of predicted hepatic fibrosis with cardiovascular 

risk factors. Methods: We conducted a cross-sectional 

study of 575 Framingham Heart Study participants with NAFLD 

based on computed tomography after excluding excess alcohol 

use. We estimated the prevalence of predicted hepatic fibrosis 

based on the aspartate aminotransferase (AST)/alanine aminotransferase 

(ALT) ratio, AST to platelet ratio index (APRI), 

and the NAFLD Fibrosis Score (NFS). Using multivariable logistic 

regression models, we examined the association between 

NAFLD with low, intermediate, or high risk for advanced fibrosis 

according to the NFS and various cardiometabolic risk factors. 

Results: Among participants with NAFLD, the prevalence 

of predicted hepatic fibrosis was 12%, 5%, and 32% based 

on the NFS, APRI, and AST/ALT ratio, respectively. Since the 

NFS predicted a high risk for advanced fibrosis most similar 

to our a priori hypothesis, we continued the analysis using 

this model to categorized participants as low, intermediate, 

or high risk for advanced fibrosis. In multivariable models, 

participants with NAFLD and a high risk for advanced fibrosis 

by the NFS had an average 6.87 mm Hg wider pulse pressure 

(95% CI 3.33-10.42 mm Hg; p=0.0002) and an increased 

odds of hypertension (OR 2.92 95% CI 1.35-6.34; p = 0.007) 

compared to those with NAFLD and low risk of fibrosis. There 

were no statistically significant differences between the systolic 

blood pressure, HDL cholesterol or triglycerides for those 

with NAFLD and a high risk of advanced fibrosis by the NFS 

compared to those with NAFLD and a low risk of advanced 

fibrosis. Conclusions: The AST/ALT ratio, APRI, and NFS give 

widely disparate predictions of hepatic fibrosis when applied 

to a community-based cohort. Participants with NAFLD and a 

high risk for advanced fibrosis based on the NFS had a wider 

pulse pressure and increased odds of hypertension. Whether 

modifying these risk factors impacts cardiovascular endpoints 

in NAFLD patients remains unknown and should be explored 

in future studies. 

Disclosures: 

Alison Pedley - Employment: Merck 

The following authors have nothing to disclose: Michelle T. Long, Joseph M. 

Massaro, Udo Hoffmann, Caroline S. Fox 

2192 

Macrophages (CD68+ Cells) and CD8+ T Cytotoxic Cells 

are Independently Associated with Advanced Fibrosis in 

Patients with Non Alcoholic Fatty Liver Disease (NAFLD) 

Azza Karrar 1 , Dinan Abdelatif 2 , Irfan Ali 2 , Yousef Fazel 1 , Pegah 

Golabi 1 , Mohamad Houry 1 , Zahra Younoszai 1 , Fanny Monge 2 , 

Munkhzul Otgonsuren 1 , Zachary D. Goodman 2 , Lakshmi Alaparthi 

2 , Zobair M. Younossi 1,2 ; 1 Betty and Guy Beatty Center for Integrated 

Research, Inova Health System, Falls Church, VA; 2 Center 

for Liver Diseases, Inova Fairfax Hospital, Falls Church, VA 

Background and Aims: Macrophages (CD68+ cells) and CD8+ 

T cytotoxic cells dominate the inflammatory infiltrate in NAFLD. 

The crosstalk between liver and adipose tissue with regard to 

inflammatory cells has not been studied in patients with NAFLD. 

We aimed to quantify macrophages and T Helpers cells in liver 

and adipose of patients with NAFLD. Methods: Liver biopsy 

samples (49) were stained with Hematoxylin and eosin (H&E) 

and Masson Trichrome for assessment of NAFLD/NASH and 

scored for fibrosis. Digitized images of CD3, CD4, CD8 and 

CD68 stained liver and adipose sections were acquired using 

The Scan scope XT. An IHC Nuclear Image Analysis algorithm 

was used to quantify the percentage of CD3+, CD4+ and 

CD8+ cells and a positive pixel count algorithm was used to 

quantify the CD68+ cells. Multivariate analysis was performed; 

p values ≤ 0.05 were considered to be significant. Results: The 

study cohort (N=49) included 33 patients with NASH NAFLD 

[Median(IQR) Age=49.00(42.00,55.00); Male=11(33%); 

White=25(75.8%); BMI(Kg/m 2 =49.22(40.27,54.67)], 

and 16 non NASH NAFLD [Age=45.00(31.00,51.50); 

Male=4(25.0%); White=14(87.5%); BMI Kg/ 

m 2 =44.53(42.23,51.49)]. In univariate analysis, percentages 

of CD3+ cells [Odds Ratio (OR): 1.44(1.02, 2.04), 

p=0.041], CD8+ cells [OR: 1.29(1.06, 1.58), p=0.011], and 

CD68+ cells [OR: 6.39 (1.11, 36.9), p=0.038] in the liver 

are significantly associated with higher risk for NASH NAFLD. 

Additionally, a greater percentage of CD8+ T cells [OR: 

1.33(1.08-1.64), p=0.006] in liver is correlated with higher 

risk for pericellular fibrosis, while in adipose, a greater percentage 

of CD3+ cells [OR: 0.67 (0.49-0.92), p=0.012] and 

CD8+ cells [OR: 0.78 (0.60-1.01), p=0.058] are correlated 

with lower risk of pericellular fibrosis. In liver, it is also found 

that a greater percentage of CD8+ cells [OR: 1.33(1.10-1.59), 

p=0.002] and CD68+ cells [OR: 5.87(1.38-25.1), p=0.017] 

are associated with higher risk of advanced fibrosis. In multivariate 

analysis, in the liver, a greater percentage of CD8+ 

cells [OR: 1.28 (1.04-1.58), p=0.020) are independently 

associated with higher risk for pericellular fibrosis and greater 

percentage of CD68+ cells [OR: 7.37 (1.58-34.4), p=0.011] 

are independently associated with higher risk of advanced 

fibrosis. Conclusion: We show that CD8+ T cells are strongly 

associated with pericellular fibrosis while CD68+ cells are 

strongly associated with advanced fibrosis. Therefore macrophages 

and CD8+ cells are key cells in NAFLD progression. 

Disclosures: 






The following authors have nothing to disclose: Azza Karrar, Dinan Abdelatif, 

Irfan Ali, Yousef Fazel, Pegah Golabi, Mohamad Houry, Zahra Younoszai, 

Fanny Monge, Munkhzul Otgonsuren, Lakshmi Alaparthi


2193 

Usefulness of mac-2 binding protein glycosylation isomer 

(M2BPGi) for advanced fibrosis and hepatocellular 

carcinoma in patients with non-alcoholic fatty liver disease 

Erina Kumagai 1,2 , Masaaki Korenaga 1 , Keiko Korenaga 1 , Misuzu 

Ueyama 1,2 , Masatoshi Imamura 1 , Masaya Sugiyama 1 , Kazumoto 

Murata 1 , Naohiko Masaki 1 , Tatsuya Kanto 1 , Sumio Watanabe 2 , 

Masashi Mizokami 1 ; 1 The research center for hepatitis and Immunology, 

National Center of Global Health and Medicine (NCGM) 

at Kohnodai, Ichikawa, Japan; 2 Department of Gastroenterology, 

Juntendo University School of Medicine, Bunkyo-ku, Hongo, Japan 

Background and Aim: Evaluation of liver fibrosis in non-alcoholic 

fatty liver disease (NAFLD) patients is important for 

identifying those who may progress to liver cirrhosis and hepatocellular 

carcinoma (HCC). Although liver biopsies are the 

gold standard for diagnosing NAFLD-associated liver fibrosis, 

there is an urgent need for a non-invasive method for estimating 

the stage of liver fibrosis in NAFLD patients.The serum 

M2BPGi value using a glycan-based immunoassay may provide 

an accurate and reliable method for assessing the liver 

fibrosis stage in not only chronic hepatitis C patients but also 

NAFLD patients; however, there is no data regarding its clinical 

usefulness and the association with HCC. The aim of this study 

was to assess the diagnostic performances of M2BPGi comparison 

with liver stiffness and Fib4-index for the diagnosis of 

advanced liver fibrosis and HCC in patients with NAFLD. Methods: 

A total of 358 consecutive patients (mean Age [ys]:63, 

male [%]:43, BMI [kg/m2]:26.3, and HCC n [%]: 25[7]) 

with NAFLD who underwent ultrasonography were prospectively 

enrolled in this study. M2BPGi (COI), Fib-4 index, and 

liver stiffness by FibroScan (kPa) were measured on the same 

day. Advanced fibrosis group was determined by cut of level 

in each marker from previous reports. (M2BPGi>1.46 COI, 

Fib4 >2.67, and FibroScan>9.8kPa) Liver stiffness measurement 

(LSM) failure was defined as zero valid shots; unreliable 

examination was defined as


2195 

Combined effects of the prosteatotic TM6SF2 and 

PNPLA3 mutations on NALFD severity: multicentre biopsy-based 

study 

Marcin Krawczyk 9,1 , Monika Rau 2 , Jörn Schattenberg 3 , Heike Bantel 

4 , Anita Pathil 5 , Münevver Demir 6 , Johannes Kluwe 7 , Tobias 

Boettler 8 , Frank Lammert 9 , Andreas Geier 2 ; 1 Laboratory of Metabolic 

Liver Diseases, Department of General, Transplant and Liver 

Surgery, Medical University of Warsaw, Warsaw, Poland; 2 Division 

of Hepatology, Department of Medicine II, University Hospital 

Wuerzburg, Wuerzburg, Germany; 3 I. Department of Medicine, 

University Medical Center Mainz, Johannes Gutenberg University, 

Mainz, Germany; 4 Department of Gastroenterology, Hepatology 

and Endocrinology, Hannover Medical School, Hannover, 

Germany; 5 Department of Internal Medicine IV, Gastroenterology 

and Hepatology, University of Heidelberg, Heidelberg, Germany; 

6 Clinic for Gastroenterology and Hepatology, University Hospital 

of Cologne, Cologne, Germany; 7 I. Department of Medicine, Hamburg 

University Medical Center, Hamburg, Germany; 8 Department 

of Medicine II, University Hospital Freiburg, Freiburg, Germany; 

9 Department of Medicine II, Saarland University Medical Center, 

Homburg, Germany 

Introduction: The PNPLA3 (adiponutrin) mutation p.I148M 

represents the common genetic risk factor for non-alcoholic 

fatty liver disease (NAFLD) and progressive liver fibrosis. Lately 

a second prosteatotic variant, TM6SF2 p.E167K, has been 

detected (Nat Genet 2014). It remains unclear if this mutation, 

alike the PNPLA3 p.I148M variant, increases the risk of 

liver fibrosis. In the current study we analyzed the combined 

effects of these variants on NAFLD severity in a large cohort 

of patients recruited at eight German tertiary referral centres. 

Patients and methods: After exclusion of acute or chronic liver 

diseases other than NAFLD, 514 patients (age 16 – 88 years, 

239 men) were included. In 309 patients liver biopsies were 

performed, which were examined by pathologists blinded to 

genotyping results. PCR-based assays were used to genotype 

the PNPLA3 (rs738409) and TM6SF2 (rs58542926) variants. 

Results: The genotype frequencies of the PNPLA3 p.I148M 

([CC] = 215, [CG] = 223, [GG] = 76) and the TM6SF2 

p.E167K ([CC] = 411, [CT] = 94, [TT] = 9) mutations did not 

deviate from Hardy-Weinberg equilibrium. One copy of the 

prosteatotic PNPLA3 and TM6SF2 alleles was detected in 58% 

and 20% of NAFLD patients, respectively. Patients carrying 

the PNPLA3 p.I148M or TM6SF2 p.E167K risk alleles had 

significantly (both P < 0.01) increased serum ALT and AST 

activities. The PNPLA3 risk genotype [GG] (OR = 2.15; 95% 

CI 1.21 - 3.87, P = 0.007), but not the TM6SF2 genotype was 

more frequent in individuals scheduled for liver biopsy. Among 

biopsied individuals, a total of 149 presented with hepatic steatosis 

grades S2 or S3, and fibrosis stage > 1 was detected in 

77 patients. The TM6SF2 variant increased the risk of developing 

steatosis grade S2 - S3 (OR = 1.52, P = 0.04) but did not 

affect fibrosis stage. The PNPLA3 genotype was, in turn, associated 

with both steatosis grades S2 - S3 (OR = 1.94, P < 0.001) 

and increased hepatic fibrosis (OR = 2.24, P < 0.001). In 

patients with the PNPLA3 genotype [GG], the presence of variant 

TM6SF2 further increased serum aminotransferase activities 

(both P < 0.05) and tended to aggravate hepatic steatosis (P 

= 0.09). Conclusions: Our results demonstrate that the PNPLA3 

and TM6SF2 variants are associated with increased liver injury 

as reflected by serum surrogate and histopathological markers. 

The TM6SF2 variant seems to modulate predominantly hepatic 

fat accumulation, while the PNPLA3 mutation confers risk of 

increased steatosis and fibrosis. 

Disclosures: 

The following authors have nothing to disclose: Marcin Krawczyk, Monika Rau, 

Jörn Schattenberg, Heike Bantel, Anita Pathil, Münevver Demir, Johannes Kluwe, 

Tobias Boettler, Frank Lammert, Andreas Geier 

2196 

Weight Loss Results in Significant Improvements in 

Quality of Life for Patients with Nonalcoholic Fatty Liver 

Disease 

Elliot B. Tapper, Michelle Lai; Gastroenterology, Beth Israel Deaconess 


Background: Nonalcoholic Fatty Liver Disease (NAFLD) is 

highly prevalent and associated with decreased quality of life 

(QOL). Longitudinal QOL data are lacking. Methods: We prospective 

enrolled patients with NAFLD from 2009-2014. All 

patients received a liver biopsy, transient elastography, lifestyle 

assessment, blood tests and QOL tools including the Chronic 

Liver Disease Questionnaire (CLDQ), a validated health-related 

quality of life measurement. All patients were followed 

for 6 months at which time the patients were re-examined and 

re-administered the CLDQ. Our primary outcome was overall 

QOL and our principle exposure variable was at least a 5% 

decrease in weight. Results: 151 patients were followed for 6 

months with complete biochemical and QOL data. The cohort 

included 91 (60%) men, aged 51.5 + 12.6 years, 46 (30%) 

of whom were diabetic. 30 (21%) had advanced fibrosis or 

cirrhosis on biopsy and 67 (47%) had nonalcoholic steatohepatitis 

(NASH), as defined by a NAFLD activity score (NAS) > 

4. Overall, 47 (31%) patients achieved at least a 5% reduction 

in weight. Neither advanced fibrosis (20% vs 22%) nor NASH 

(44% vs 54%, p = 0.28) were predictive of weight loss. QOL 

improved significantly for patients who achieved at least a 

5% reduction in weight, especially in nondiabetic patients, 

those with NASH, and without significant fibrosis. The cohort’s 

baseline total CLDQ value was 5.6 (4.8 – 6.2). Those who 

achieved at least a 5% reduction in weight had a 0.45 (95% 

CI:0.24 – 0.66, p


coholic Fatty Liver Disease (NAFLD), where the average BMI 

is higher than in Europe and Asia, is limited. We examined 

predictors of liver stiffness measurement (LSM) change and 

the effect of a combined testing strategy employing NAFLD 

Fibrosis Score (NFS) and VCTE. Methods: A prospective cohort 

of 161 biopsy-proven NAFLD patients evaluated with VCTE 

using an M probe and NFS at baseline and a repeat VCTE 

at 6 months. Unreliable VCTE readings were defined as a 

failure to obtain 10 valid measurements or an interquartile 

range (IQR) > 30%. Results: 121 (75.2%) patients had reliable 

VCTE. The mean body mass index (BMI) of the total cohort 

was 33.5 km/m 2 ; 32.2 and 37.2 in those with reliable and 

unreliable VCTE (p < 0.0001). The optimal BMI cutoff for an 

unreliable exam is BMI 37 kg/m 2 (odds ratio 6.76 95% CI 

(3.06 – 15.4). The area under the receiver operating curve 

(AUROC) for the detection of F3-F4 fibrosis by VCTE is 0.92 

(95% CI: 0.85–0.96) with an optimal LSM cutoff of 9.9 kPa 

(sensitivity 95%, specificity 77%). Conversely, the AUROC for 

the association between NFS and advanced fibrosis for the 

whole cohort was 0.77 (95% CI:0.63–0.97). In the detection 

of F3-F4 fibrosis in patients with reliable VCTE, the AUROC for 

VCTE is superior to that of NFS (0.92 vs. 0.77, p=0.01). VCTE 

led to a lower F3-F4 fibrosis misclassification rate (13% versus 

17%) A low risk VCTE result was obtained for 63 patients with 

F0-F2 fibrosis. However, 13 of these patients had indeterminate 

NFS; a combined strategy correctly classifies fewer (50) 

low-risk patients. The median LSM improvement after 6 months 

was 11.1% (IQR= -13.0% - 37.8%) and 37 patients (45%) 

experienced a 20% improvement. A 5% weight loss was associated 

with improved LSM (p = 0.009). Conclusion: Reliable 

VCTE exams are useful to rule out advanced fibrosis in the U.S. 

NAFLD patients. However, 25% have unreliable studies using 

an M probe. Weight loss improves liver stiffness in the short 

term, likely by addressing steatosis and not fibrosis. Further 

data using the XL probe and adjustment for hepatic steatosis 

will be important. 

Disclosures: 

The following authors have nothing to disclose: Elliot B. Tapper, Michelle Lai 

2198 

Functional inhibition of spleen tyrosine kinase ameliorates 

different stages of alcoholic liver disease in mice 

Terence N. Bukong, Arvin Iracheta-Vellve, Aditya Ambade, Karen 

Kodys, Gyongyi Szabo; Medicine, University of Massachusetts 

Medical School, Worcester, MA 

Background: The spectrum of alcoholic liver disease (ALD) 

includes steatosis triggered by acute alcohol binge drinking, 

chronic alcoholic hepatitis and cirrhosis, the latter a leading 

cause of mortality with limited therapies available. Because 

alcohol targets different signaling pathways in the liver, identification 

of a master regulatory target capable of modulating 

multiple signaling processes is attractive. The aim of this study 

was to evaluate the role of spleen tyrosine kinase (SYK), a 

non-receptor tyrosine kinase, given its central modulatory role 

of multiple pro-inflammatory signaling pathways previously 

identified in the pathomechanism of ALD. Methods: Three different 

models of ALD were assessed in mice: 3-day binge, 

10-day acute-on-chronic and 5 weeks chronic alcohol Lieber-DeCarli 

diet. Some mice received daily dose of SYK inhibitor 

(R406) [5-10mg/kg]. Liver and serum samples were assessed 

by western blot, ELISA, EMSA, qPCR and histology. Results: 

Liver Syk protein expression was increased by acute-on-chronic 

and chronic but not acute alcohol administration. However, 

we found a significant elevation of activated Syk and its downstream 

targets, phosphorylated(p)-SYK Y525/526 , p-ERK1/2 and 

p-p38, in livers of alcohol-fed mice compared to controls in 

all 3 models. In all models, alcohol administration led to liver 

injury and inflammation demonstrated by significant increases 

in ALT, p-ERK1/2, NF-κB binding, TNF-α, MCP1, and IL-1β protein 

expression compared to controls. In vivo administration of 

a functional SYK inhibitor attenuated alcohol-induced hepatic 

injury and inflammation indicated by significantly reduced ALT, 

p-ERK1/2, p-p38, NF-κB binding, TNF-α, MCP1, E-Selectin, 

IL-1β expression compared to vehicle treated ethanol-fed mice. 

The protective effect of SYK inhibition on liver inflammation 

was present in all 3 models; specifically, neutrophil activation 

markers, Ly6G, MPO and E-Selectin, were decreased in the 

acute-on-chronic model and macrophage activation (F4/80 

and CD68) was attenuated in chronic alcohol feeding. Furthermore, 

SYK inhibition resulted in a significant decrease in 

hepatic steatosis indicated by Oil Red-O staining, triglyceride 

levels and increases in UCP1 and PRDM16 expression in the 

liver suggesting that SYK-dependent signaling is involved in 

alcoholic steatosis at different levels, including lipogenesis and 

lipid metabolism. Conclusions: Our data demonstrate a novel 

functional role of SYK in modulating liver inflammation and 

steatosis at different stages of alcoholic liver damage. This 

study highlights SYK as a potential multifunctional therapeutic 

target to suppress inflammation and steatosis in alcoholic liver 

disease. 

Disclosures: 

The following authors have nothing to disclose: Terence N. Bukong, Arvin Iracheta-Vellve, 

Aditya Ambade, Karen Kodys, Gyongyi Szabo 

2199 

The glycosylation marker M2BPGi predicts the development 

of hepatocellular carcinoma in nonalcoholic 


Miwa Kawanaka 1 , Hideyuki Hyogo 2 , Masahiko Koda 3 , Toshihide 

Shima 5 , Yuichi Hara 6 , Hiroshi Tobita 4 , Ken Nishino 1 , Akira Hiramatsu 

2 , Shuichi Sato 4 , Kazuaki Chayama 2 , Hirofumi Kawamoto 1 , 

Takeshi Okanoue 5 , Keisuke Hino 6 ; 1 General Intrenal medicine2, 

Kawasaki-Hostital,Kawasaki Medical school, Okayama-city, 

Japan; 2 Medicine and Molecular Sciences,Graduate School of 

Biomedical Sciences, Hiroshima University, Hiroshima, Japan; 

3 Department of Multidisciplinary Internal Medicine, Tottori University 

School of Medicine, Yonago, Japan; 4 Gastroenterology 

and Hepatology, Shimane University, School of Medicine, Izumo, 

Japan; 5 Saiseikai Suita Hospital, Suita city, Japan; 6 Hepatology 

and Pancreatology, Kawasaki Medical School, Kurashiki city, 

Japan 

[Objectives] The number of non-B non-C hepatocellular carcinoma 

(HCC) cases has increased rapidly in recent years, 

and the increasing in the incidence of nonalcoholic steatohepatitis 

(NASH) has been an associated factor. Therefore, 

non-invasive methods are important to detect NASH patients 

with a high carcinogenesis risk.The present study examined 

the utility of M2BPGi as an indicator of liver carcinogenesis 

in NASH patients.[Subjects and methods] In this multicenter 

study, 63 patients diagnosed as NASH on the basis of hepatic 

histological findings and detected to have HCC (NASH-HCC) 

were compared with 280 control patients who, on the basis of 

liver biopsy findings, had not developed HCC for 9.8 years 

after a NASH diagnosis (NASHnon-HCC) in terms of age, 

sex, body mass index, ALT, AST, GTP, AST/ALT ratio, platelet 

count, homeostatic assessment model of insulin resistance, 

iron, ferritin, and high-sensitivity C-reactive protein, hyaluronic 

acid, type IV collagen 7S, and P-III-P, glycosylation markers 

(M2BPGi), and fibrosis-4 (FIB4) index, which contribute to HCC 

development.[Results] Patients in the NASH-HCC group were


older, more likely to be male, and had a higher incidence of 

diabetes than those in the NASH non-HCC group. The NASH- 

HCC group had more patients with advanced liver fibrosis. The 

NASH-HCC group had a lower ALT level, platelet count, and 

albumin level and significantly higher fibrosis marker levels, 

ALT/AST ratio, M2BPGi, and FIB4 index than the NASH non- 

HCC group. M2BPGi was 1.1 in the NASH non-HCC group 

and 3.4 in the NASH HCC group (P < 0.0001). M2BPGi 

levels were examined at fibrosis stage 3 (1.1 vs. 2.1, P < 

0.005) and 4 (2.6 vs. 4.6, P < 0.02); HCC developed in 

patients with higher M2BPGi levels. In multivariate analysis, 

age, sex, fibrosis stage, and M2BPGi were independent factors 

contributing to NASH-HCC. An age of ≥67 years (odds 

ratio: 10.5; 95% CI [4.5–26.2]), male sex (odds ratio: 6.935; 

95% CI [2.8–17.2]), fibrosis stages 2–4 (stage 2: odds ratio: 

2.4 [0.7–10.4]; stage 3: odds ratio: 2.6 [0.7–10.8]; stage 

4: odds ratio: 18.3 (4.9–82), P < 0.0001) versus stages 0-1, 

and M2BPGi levels of 1.25–2.66 (odds ratio: 2.0 [0.8–5]) 

and ≥2.67 (odds ratio 6.3 [1.7–24.2]) were associated with 

a higher risk of carcinogenesis. Thus, the risk was markedly 

elevated in patients with M2BPGi levels of ≥2.67. The rate of 

carcinogenesis at 10 years was 60% in patients with M2BPGi 

levels ≥2.67 and 15% in those with M2BPGi levels ≤2.67. 

[Conclusions] HCC risk was higher in those with non-alcoholic 

fatty liver disease who were men, aged ≥67 years, or had 

M2BPGi levels ≥2.67. Rourine follow-up using imaging tests is 

essential in these patients. 

Disclosures: 








The following authors have nothing to disclose: Miwa Kawanaka, Hideyuki 

Hyogo, Masahiko Koda, Toshihide Shima, Yuichi Hara, Hiroshi Tobita, Ken 

Nishino, Akira Hiramatsu, Shuichi Sato, Hirofumi Kawamoto, Takeshi Okanoue, 

Keisuke Hino 

2200 

Improvement in Hepatic Steatosis in Nonalcoholic Steatohepatitis 

in Response to Synbiotic Supplementation 

Silvia M. Ferolla, Claudia A. Couto, Geyza N. Armiliato, Luciana 

C. Silva, Erika C. Lima, Quelson C. Lisboa, Flaviano S. Martins, 

Maria de Lourdes A. Ferrari, Aloisio S. Cunha, Cristiano A. 

Pereira, Teresa C. Ferrari; Federal University of Minas Gerais, Belo 

Horizonte, Brazil 

Background: Nonalcoholic fatty liver disease (NAFLD) is one 

of the most prevalent chronic liver diseases in the world, which 

can progress to a severe form of nonalcoholic steatohepatitis 

(NASH), cirrhosis and even hepatocellular carcinoma. Oral supplementation 

with a synbiotic has been proposed as an effective 

treatment of NAFLD as these agents influence the gut-liver 

axis improving inflammatory parameters. Objective: We aim 

to evaluate the effects of supplementation with a synbiotic on 

the grade of hepatic steatosis, intestinal bacterial overgrowth, 

and serum levels of lipopolysaccharide (LPS) and adiponectin 

in NASH patients. Design: In a randomized, controlled clinical 

trial, 50 patients with biopsy-proven NASH were supplemented 

twice daily for three months with a synbiotic or not receive any 

supplementation. Both groups were advised to follow an energy-balanced 

diet and physical activity recommendations. The 

anthropometric and metabolic parameters, grade of steatosis 

(evaluated by magnetic resonance [MR] techniques), glucose 

hydrogen breath test, and levels of LPS and adiponectin were 

evaluated before and after the treatment. Results: At the end of 

the study, the following significant differences were observed in 

the synbiotic group when compared to the control individuals: 

decrease in the hepatic proton density fat fraction measured 

by MR (-3.4%; p=0.027); loss of body weight (-1.3kg; 1.5%; 

p=0.06); reduction in BMI (-0.4kg/m 2 ; 1.2%; p=0.005) and 

reduction in the waist circumference measurement (-1.9cm; 

1.8%;p=0.001). Food consumption (total daily calorie intake) 

was not different between groups. Conclusions: Synbiotic supplementation 

in addition to nutritional counseling seems superior 

to nutritional counseling alone for the treatment of NAFLD, 

as the intervention group presented attenuation of steatosis, 

and reduction of body weight, BMI and waist circumference. 

Whether these effects will be sustained with longer treatment 

durations remains to be determined. 

Disclosures: 

The following authors have nothing to disclose: Silvia M. Ferolla, Claudia A. 

Couto, Geyza N. Armiliato, Luciana C. Silva, Erika C. Lima, Quelson C. Lisboa, 

Flaviano S. Martins, Maria de Lourdes A. Ferrari, Aloisio S. Cunha, Cristiano A. 

Pereira, Teresa C. Ferrari 

2201 

Cross-sectional and longitudinal analysis of advanced 

MRI-derived measures of liver fat and volume in the 

treatment of nonalcoholic steatohepatitis: A secondary 

analysis of an RCT 

Steven C. Lin 1 , Elhamy Heba 2 , Ricki Bettencourt 1,3 , Claude B. 

Sirlin 2 , Rohit Loomba 1,3 ; 1 NAFLD Translational Research Unit, Division 

of Gastroenterology, University of California, San Diego, La 

Jolla, CA; 2 Liver Imaging Group, Department of Radiology, University 

of California, San Diego, La Jolla, CA; 3 Division of Epidemiology, 

Department of Family and Preventive Medicine, University of 

California, San Diego, La Jolla, CA 

Background: Magnetic resonance imaging (MRI)-derived measures 

of liver fat and volume are emerging as accurate, non-invasive, 

and validated imaging biomarkers in patients with 

biopsy-proven nonalcoholic steatohepatitis (NASH). However, 

little is known about the changes in these measures longitudinally, 

and their association with each other in a NASH clinical 

trial. Aim: This study aims to examine the cross-sectional 

relationships between MRI-derived proton density fat fraction 

(PDFF, %), total liver volume (TLV, mL), and total liver fat index 

(TLFI, % mL) at two time points in a NASH randomized clinical 

trial. Methods: This is a secondary analysis of a randomized, 

double-blind, placebo-controlled trial (MOZART) in which 50 

patients with biopsy-proven NASH were randomized to either 

oral ezetimibe 10mg daily (n = 25) vs. placebo (n = 25) over 

24 weeks for the treatment of NASH (Loomba et al. Hepatology 

2014). Baseline and post-treatment anthropometrics, biochemical 

profiling, MRI, and liver biopsies were obtained. PDFF, TLV, 

and TLFI were obtained and calculated from MRI scans captured 

at these two time points. Results: At baseline, mean PDFF 

correlated strongly with TLFI (Spearman’s ρ = 0.94, n = 45, 

P


trial, PDFF and TLV were both strongly correlated with TLFI. An 

increase in hepatic steatosis (as measured by PDFF) is associated 

with an increase in hepatomegaly, as measured by TLV. 

Our findings indicate that MRI-derived measures of liver fat and 

volume can be used both cross-sectionally and longitudinally as 

imaging biomarkers in a NASH trial. The varied strengths of 

correlations between these MRI-derived measures suggest that 

the use of all three measures provides a more comprehensive 

assessment of changes in liver fat burden and size, rather than 

the use of only one measure in isolation. (Clinicaltrials.gov 

NCT01766713) 

Disclosures: 







The following authors have nothing to disclose: Steven C. Lin, Elhamy Heba, 

Ricki Bettencourt 

2202 

Importance of reduction in serum alanine aminotransferase 

and the effect of PNPLA3 rs738409 

polymorphism for preventing the histological disease 

progression of nonalcoholic steatohepatitis in Japanese 

patients: a multi-center, retrospective study 

Yuya Seko 1 , Yoshio Sumida 1 , Yuichiro Eguchi 2 , Hideyuki Hyogo 3 , 

Saiyu Tanaka 4 , Kohjiroh Mori 4 , Keizo Anzai 2 , Kazuaki Chayama 

5 , Yoshito Itoh 1 ; 1 Kyoto Prefectural University of Medicine, 

Kyoto, Japan; 2 Division of Hepatology and Internal medicine, 

Saga Univercity, Saga, Japan; 3 JA Hiroshima General Hospital, 

Hiroshima, Japan; 4 Center for Digestive and Liver Diseases,, Nara 

City Hospital, Nara, Japan; 5 Department of Gastroenterology and 

Metabolisim,, Hiroshima University Hospital, Hiroshima, Japan 

BACKGROUND & AIMS: Some environmental factors and 

genetic variation were reported to associate with histological 

progression in patients with nonalcoholic fatty liver disease 

(NAFLD). Our aim is to determine predictors, including PNPLA3 

rs738409 (encoding the I148M variant), of histological progression 

in Japanese patients with biopsy-proven NASH. 

METHODS: This multi-center retrospective cohort study enrolled 

140 patients with NASH who underwent serial liver biopsies. 

Histological evaluation included NASH activity score (NAS) 

and liver fibrosis. The median interval between initial and second 

liver biopsies was 900 days. ALT response was defined 

as a decrease to ≤40 U/L or by ≥30% of baseline. RESULTS: 

Of 140 patients, NAS was ameliorated in 50.7%, deteriorated 

in 20.0%, and remained unchanged in 29.3%. Liver fibrosis 

was improved in 22.9% of patients, progressed in 22.9%, and 

remained stable in 54.3%. The PNPLA3 rs738409 genotype 

frequencies were CC 0.29, CG 0.42, and GG 0.28. Multivariate 

analysis identified woman (hazard ratio [HR], 5.36; 

p=0.021) and ALT non-response (HR, 6.62; p=0.008) as predictors 

of deterioration of NAS. ALT non-response (HR, 2.60; 

p=0.043) was also selected as a predictor of progression of 

liver fibrosis. The average annual rate of fibrosis was -0.06 

stages/year in ALT responders, increasing to 0.13 stages/year 

in ALT non-responders. Carriage of G allele was associated 

with deterioration of NAS (p=0.045), but not associated with 

fibrosis progression. ALT responders were found in 78.1% of 

CC homozygotes, 66.7% of CG and 53.3% of GG. The ALT 

responders frequencies in PNPLA3 CC was 78.1%, in CG was 

66.7%, and in GG was 53.3%. Carriage of G allele was tend 

to associated with ALT non-responders (p=0.070). CONCLU- 

SIONS: A lack of reduction in serum ALT level was a predictor 

for histological progression in patients with NASH. Serum ALT 

should be strictly controlled to prevent liver histological progression 

in patients with NASH, and ALT response was associated 

with PNPLA3 rs738409 G allele. 

Disclosures: 













The following authors have nothing to disclose: Yuya Seko, Yoshio Sumida, 

Yuichiro Eguchi, Hideyuki Hyogo, Saiyu Tanaka, Kohjiroh Mori, Keizo Anzai 

2203 

First degree relatives of subjects with type 2 Diabetes 

Mellitus get early and severe NASH related cirrhosis 

Ajeet S. Bhadoria 1 , Chandan K. Kedarisetty 2 , Ankit Bhardwaj 1 , 

Guresh Kumar 1 , Tanmay S. Vyas 2 , Jaya Benjamin 3 , Varsha 

Shasthry 3 , Manoj Kumar 2 , Shiv K. Sarin 2 ; 1 Clinical Research, Institute 

of Liver and Biliary Sciences, New Delhi, India; 2 Hepatology, 

Institute of Liver and Biliary Sciences, New Delhi, India; 3 Clinical 

Nutrition, Institute of Liver and Biliary Sciences, New Delhi, India 

Background and Aims: Familial aggregation of type 2 diabetes 

mellitus (T2DM) and NAFLD is well documented. However, 

there is scarcity of data on such association with NASH related 

cirrhosis. We investigated the influence of family history (FH) of 

T2DM on age at diagnosis and severity at presentation among 

NASH related cirrhotics. Patients and Methods: In a cross-sectional 

study, all NASH related cirrhotics presenting to Hepatology 

Department of ILBS, New Delhi from Jan 2014-May 

2015 were included. Other etiologies, like alcoholic, viral, 

autoimmune, cholestatitc liver diseases and inherited metabolic 

disorders were excluded. FH, demographic characteristics, 

medical history data, anthropometrical measurements and laboratory 

data were recorded. FH for T2DM was defined as at 

least one first degree relative having T2DM. Results: Out of 

1,033 NASH related cirrhotics (71.8% males) with mean age 

54.7 (±11.1) years, 549 (53.1%) had FH for T2DM (


MTs had HCC (7.0% Vs 3.1%, p=0.004) at presentation. 

Conclusions: A significant number (52.7%) of NASH related 

cirrhotics had FH for T2DM. Importantly, FH of T2DM constitutes 

a high risk group for early and severe cirrhosis. Community 

based research to further substantiate this hypothesis and 

health education strategies to disseminate relevant awareness, 

is a prime requisite. 

Disclosures: 

The following authors have nothing to disclose: Ajeet S. Bhadoria, Chandan K. 

Kedarisetty, Ankit Bhardwaj, Guresh Kumar, Tanmay S. Vyas, Jaya Benjamin, 

Varsha Shasthry, Manoj Kumar, Shiv K. Sarin 

2204 

Non-alcoholic Fatty Liver Disease (NAFLD) Is Associated 

with Impairment of Health Related Quality of Life 

(HRQoL): Data from US Population 

Munkhzul Otgonsuren 2 , Rebecca Cable 2 , Sean C. Felix 2 , Patrick 

W. Austin 2 , Yun Fang 2 , Zobair M. Younossi 2,1 ; 1 Center For Liver 



Inova Health System, Falls Church, VA 

Background and Aim: NAFLD is an important cause of chronic 

liver disease (CLD). Our aim was to assess the impact of NAFLD 

on patients’ HRQoL in the US adult population. Methods: 

National Health and Nutrition Examination Survey (NHANES 

2001-2011) was used to identify adults with NAFLD [Fatty Liver 

Index (FLI)>60 in the absence of other causes of liver disease, 

chronic diseases and excessive alcohol use]. Subjects without 

any of these conditions were considered as healthy controls. 

Patients with HCV RNA(+) were considered as HCV-controls. 

All patients completed HRQoL-4 questionnaire: (1) Component 

1 (C1) rate health status as poor, fair, good, very good, or 

excellent; (2) C2: thinking about your physical health, which 

includes physical illness and injury, for how many days during 

the past 30 days was your physical health not good; (3) C3: 

thinking about your mental health, which includes stress, 

depression, and problems with emotions, for how many days 

during the past 30 days was your mental health not good; and 

(4) C4: about how many days did poor physical/mental health 

keep him/her from doing your usual activities, such as selfcare, 

work, or recreation. Linear regression models adjusted 

for age, gender, race, and BMI was used to assess associations. 

Results: 9,661 NHANES participants (3,333 NAFLD, 

346 HCV+ and 5,982 controls). The proportion of subjects 

rating their health as “fair” or “poor” was higher in NAFLD 

(20%) than healthy controls (10%). However, 30% of HCV 

reported poor to fair health (all p


= 4.80 (2.88-9.69)] are independent predictors of mortality 

(all p


ciated with HCC recurrence in ALD-HCC. Conclusion: Although 

NAFLD-HCC and ALD-HCC differ in several characteristics, the 

clinical course of these two diseases is similar. Both have a 

high recurrence rate as with other types of HCC. Expression of 

the tumor markers DCP and AFP differed between NAFLD-HCC 

and ALD-HCC patients and might be useful for predicting HCC 

recurrence. 

Disclosures: 

The following authors have nothing to disclose: Tomomi Kogiso, Etsuko 

Hashimoto, Hideyuki Hyogo, Tomoaki Nakajima, Masafumi Ono, Takumi Kawaguchi, 

Koichi Honda, Yuichiro Eguchi, Yuichi Nozaki, Miwa Kawanaka, Kento 

Imajo, Yoshio Sumida, Yoshihiro Kamada, Hideki Fujii, Yasuaki Suzuki, Katsutoshi 

Tokushige 

2208 

Diacylglycerol acyltransferase 1 inhibition, as a metabolic 

regulator: clinical benefits of pradigastat in 

patients with non-alcoholic fatty liver disease 

Kenneth Cusi 1 , Arun J. Sanyal 2 , Samir Patel 3 , Melanie Wright 4 , 

Changlu Liu 3 , Deborah L. Keefe 3 ; 1 Division of Endocrinology, 

Diabetes and Metabolism, University of Florida, Gainesville, FL; 

2 Division of Gastroenterology, Hepatology and Nutrition, Virginia 

Commonwealth University, Richmond, VA; 3 Novartis Pharmaceuticals 

Corporation, East Hanover, NJ; 4 Novartis Pharma AG, Basel, 

Switzerland 

Purpose: Non-alcoholic fatty liver disease (NAFLD) affects up 

to 80% of obese and diabetic patients. Pradigastat, a potent 

selective diacylglycerol acyltransferase 1 (DGAT1) inhibitor, 

reduces post-prandial triglycerides, glucose and insulin 

in overweight or obese patients. This study assessed the efficacy 

and safety of pradigastat on multiple metabolic parameters 

in NAFLD patients. Methods: This was a multicenter, 

randomized, double-blind, placebo-controlled 24-week study 

(NCT01811472). NAFLD patients with liver fat ≥10% were 

randomized (2:1:2) to placebo, pradigastat 5/10 mg or 

10/20 mg and up-titrated from pradigastat 5 to 10 mg (or 

10 to 20 mg) after 2 weeks. Due to the pilot nature of the 

study, the significance level was set at the 1-sided 5% (2-sided 

10%) level, and not fully powered for many of the metabolic 

parameters listed in Table 1. Results: A total of 52 patients 

were randomized, of whom 45 (87%) completed the study. 

Mean baseline BMI and waist circumference were 33.4 kg/ 

m 2 and 110.0 cm, respectively, with 80% of patients being 

obese. There was a modest decrease in body weight and waist 

circumference with pradigastat compared to placebo, with the 

differences reaching statistical significance at Weeks 4 to 8 

(5/10 mg) and up to Week 20 (10/20 mg) for weight, and 

at week 12 for waist circumference (10/20 mg) (Table 1). A 

modest decrease was also observed in abdominal subcutaneous 

and visceral fat, as well as in some metabolic parameters 

(Table 1). Pradigastat was overall well tolerated except for GI 

symptoms. Diarrhea affected 35.0%, 54.5%, and 85.7% of 

patients on placebo, pradigastat 5/10 and 10/20 mg, respectively. 

Conclusion: Pradigastat treatment for up to 24 weeks 

led to a modest reduction in body weight (1.3-1.4 kg). Both 

pradigastat groups were associated with numerically greater 

and sustained decreases in different markers of body fat compared 

to placebo. The significance of these effects needs to be 

confirmed in larger studies. 

Table 1: Adjusted mean change from baseline of metabolic 

parameter 

*significant at 2-sided 10% level vs placebo 

Disclosures: 

Kenneth Cusi - Consulting: Merck, Janssen, Pfizer, Pfizer; Grant/Research Support: 

Janssen, Novartis 






Samir Patel - Employment: Novartis Pharmaceutical 

Melanie Wright - Employment: Novartis Pharma AG 

Changlu Liu - Employment: Novartis Pharmaceuticals 

Deborah L. Keefe - Employment: Novartis Pharmaceuticals Corp 

2209 

Subjects with non alcoholic fatty liver disease (NAFLD) 

display increased visceral and pancreatic fat associated 

with worse metabolic profile 

Chiara Saponaro 1 , Melania Gaggini 1 , Chiara Rosso 2 , Emma 

Buzzigoli 1 , Fabrizia Carli 1 , Demetrio Ciociaro 1 , Lavinia Mezzabotta 

2 , Francesca Saba 2 , Andrea Marengo 2 , Maria Lorena 

Abate 2 , Riccardo Faletti 2 , Maurizio Cassader 2 , Roberto Gambino 

2 , Antonina Smedile 2 , Mario Rizzetto 2 , Elisabetta Bugianesi 2 , 

Amalia Gastaldelli 1 ; 1 CNR, Pisa, Institute of Clinical Physiology, 

Pisa, Italy; 2 Dept. of Medical Sciences, University of Turin, Division 

of Gastroenterology and Hepatology and Lab. of Diabetology, 

Turin, Italy 

Together with insulin resistance (IR), lipotoxicity and inflammation, 

patients with non-alcoholic fatty liver diseases (NAFLD) 

often show increased visceral fat (VF) that correlates with 

increased insulin and hepatic fat (HF). If VF is playing a key 

role in the progression of liver disease is not clear. Beyond 

VF and HF, fat can also accumulate in the pancreas (PF) leading 

to impaired insulin secretion and increasing the risk of 

diabetes (T2DM). The aim of this work was to measure by 

magnetic resonance (MR) the separate impact of VF, HF and 

PF on histological liver damage, lipid profile and metabolic 

alterations in non diabetic patients with biopsy proven NAFLD. 

Methods: We studied 34 non diabetic subjects with NAFLD 

(biopsy scored according to Kleiner) and 8 healthy controls 

(CT). We measured plasma concentrations of triglycerides 

(TG), free fatty acids (FFA), insulin, C-peptide, monocyte chemoattractant 

protein-1(MCP-1) and adiponectin, FFAs composition 

by chromatography mass spectrometry (GCMS), visceral, 

hepatic and pancreatic fat by MR. In addition, using tracers, 

we evaluated lipolysis and endogenous glucose production 

(EGP). We calculated indexes of IR (i.e., HOMA, Adipo-IR 

=Lipolysis x Insulin and Hep-IR= EGP x Insulin); the ratios palmitic/linoleic 

acid (16:0/18:2, an indirect index of de novo 

lipogenesis, DNL) and the saturated to unsaturated fat ratio 

(SFA/PUFA, associated with impaired lipid metabolism and 

oxidative stress). Results: NAFLD patients with fibrosis F1-4 

(n=24) had a worse metabolic profile compared to NAFLD 

with F0 (n=10) and CT, showing increased parameters of


insulin resistance, inflammation (MCP-1) and lipotoxicity (i.e. 

increased DNL index and SFA/PUFA). Visceral and hepatic 

fat were both significantly increased in NAFLD, especially 

in F1-4 vs F0 vs CT (VF=2.9±1.1 vs 2.1±0.6 kg vs 0.7±0.4 

kg p


2212 

A Meta-analytic Assessment of Worldwide Prevalence 

of Non Alcoholic Fatty Liver Disease (NAFLD) and Associated 

Co-morbidities 

Aaron B. Koenig 1 , Dinan Abdelatif 1 , Yousef Fazel 1 , Mark Wymer 1 , 

Linda Henry 1 , Zobair M. Younossi 2 ; 1 Betty and Guy Beatty Center 

for Integrated Research, Inova Health System, Falls Church, VA; 


Hospital, Falls Church, VA 

Background: NAFLD has become a major public health problem 

worldwide. Our aim was to determine the prevalence of 

NAFLD worldwide, its associated comorbidities, and prevalence 

of non-alcoholic steatohepatitis (NASH). Methods: A systematic 

literature review using PubMed, Ovid Medline and 

Web of Science was done with appropriate search terms. 

Exclusion criteria: any study for special population (morbid 

obesity) and alcohol consumption>10 g/day. Studies were 

reviewed by 3 investigators. Reporting followed the Preferred 

Reporting Items for Systematic reviews and Meta-Analyses 

(PRISMA) statement. Random-effects model univariate meta-regression 

explored high heterogeneity sources (i.e. age, gender, 

country, year of study, and study design). Results: 700+ articles 

were reviewed. After exclusion criteria, 95 studies examined, 

53 analyzed [Asia n=13, Europe n=13, Middle East 

(ME) n=3, North America (NA) n=23 and South American (SA) 

n=1]. Number of studies for prevalence of comorbidities varied 

[obesity n=19; diabetes (DM) n=35, hyperlipidemia (HL) 

n=14, hypertriglyceridemia (HTG) n=8, metabolic syndrome 

(MS) n=18, hypertension (HTN) n=26, NASH n=4]. Worldwide 

NAFLD prevalence was 21.3% (17.9%-30.5%); By continent, 

Asia- 24%(19.6%-30.5%); Europe-21%(12.7%-31.7%); 

Middle East- 31.8% (13.5%-58.2%); North America- 18.5% 

(14.3%-23.6%); SA-35.3% (27.8%-43.5%)(Figure). Rate of 

obesity was 47.4% (34.2%-60.9%); DM: 18% (13.6%-23.5%); 

HL-62.2% (45.3%-76.5%); HTG- 37.3% (25.4%-51.0%); MS- 

43.3% (29.4%-58.3%); HTN- 38.6% (32.1%-45.7%) and 

NASH 26.2% (17.3%-37.7%). Conclusion: The prevalence of 

NAFLD and NASH are high with some variation throughout 

the world. HL and MS appeared to be the most prevalent risk 

factors associated with NAFLD. 

Disclosures: 



The following authors have nothing to disclose: Aaron B. Koenig, Dinan Abdelatif, 

Yousef Fazel, Mark Wymer, Linda Henry 

2213 

Risk factors for progression of liver disease status in 

Japanese patients with non-alcoholic steatohepatitis 

Ayako Urabe 1 , Naoki Hiramatsu 1 , Tsugiko Oze 1 , Takayuki 

Yakushijin 1 , Ryoko Yamada 1 , Naoki Morishita 1 , Yuki Tahata 1 , 

Yuichi Yoshida 1 , Tomohide Tatsumi 1 , Masahide Oshita 2 , Akira 

Kaneko 3 , Eiji Mita 4 , Masahiko Tsujii 5 , Toshihiko Nagase 6 , 

Hiroyuki Fukui 8 , Kunio Suzuki 7 , Atsuo Inoue 9 , Yasuharu Imai 10 , 

Tetsuo Takehara 1 ; 1 Gastroenterology and Hepatology, Osaka University 

graduate School of Medicine, Suita, Japan; 2 Osaka Police 

Hospital, Osaka, Japan; 3 NTT West Osaka Hospital, Osaka, 

Japan; 4 National Hospital Organization Osaka National Hospital, 

Osaka, Japan; 5 Osaka Rosai Hospital, Sakai, Japan; 6 Suita 

Municipal Hospital, Suita, Japan; 7 Saiseikai Senri Hospital, Suita, 

Japan; 8 Yao Municipal Hospital, Yao, Japan; 9 Osaka General 

Medical Center, Osaka, Japan; 10 Ikeda Municipal Hospital, Ikeda, 

Japan 

Background & Aim: The incidence of hepatocellular carcinoma 

(HCC) of chronic liver disease with hepatitis C virus infection 

is decreasing by the advance of antivirus therapy, while the 

increase of HCC from non-alcoholic steatohepatitis (NASH) 

is becoming a problem, recently. However, the risk factors 

for progression of NASH are still unknown. We performed 

a longitudinal study to investigate factors associated with the 

progression of histologic fibrosis and long-term outcomes of 

NASH. Patients & Methods: We did a retrospective analysis 

of 150 Japanese patients histologically diagnosed with NASH 

from March 1998 to March 2009 at the Osaka University 

Hospital and other institutions participating in the Osaka Liver 

Forum. All patients were Japanese. The patient characteristics 

at baseline were: mean age, 55.7±15.5 y.o.; male/female, 

86/64; BMI>25kg/m 2 , 104 (69%); ALT level, 113.6±82.8 

U/L; platelet count, 20.5±7.8 /μL. Concurrent diseases were: 

dyslipidemia, 77 (51%); hypertension, 56 (37%); diabetes, 49 

(33%). Histological appearance according to Brunt classification 

was: grade 1/2/3, 57(38%)/74(49%)/16(11%); stage 

1/2/3/4, 49(33%)/42(28%)/39(26%)/17(11%). Risk factors 

associated with the incidence of decompensated cirrhosis 

(varix, ascites, hepatic encephalopathy), HCC and liver-related 

death were examined for 134 patients who were followed up. 

The mean observation period was 72.9 months±41.2 months. 

The Kaplan-Meiyer method and Cox proportional-hazards 

model were used. Results: Significant factors associated with 

advanced fibrosis at baseline were diabetes (p=0.016), Brunt 

grade (p


Disclosures: 





The following authors have nothing to disclose: Ayako Urabe, Naoki Hiramatsu, 

Tsugiko Oze, Takayuki Yakushijin, Ryoko Yamada, Naoki Morishita, Yuki 

Tahata, Yuichi Yoshida, Tomohide Tatsumi, Masahide Oshita, Akira Kaneko, Eiji 

Mita, Masahiko Tsujii, Toshihiko Nagase, Hiroyuki Fukui, Kunio Suzuki, Atsuo 

Inoue, Yasuharu Imai 

2214 

Concomitant Non-Alcoholic Fatty Liver Disease Drives 

Progression of Overall Liver Disease More Than Chronic 

Hepatitis B Alone in Chinese-Americans 

Rouchelle D. dela Cruz 1 , Lan Sun Wang 1 , Anthony Ng 3 , Rene S. 

Eng 3 , Lauren M. Eng 3 , Neil D. Theise 1,2 ; 1 Division of Digestive 

Diseases, Mount Sinai Beth Israel Medical Center, New York, NY; 

2 Department of Pathology, Mount Sinai Beth Israel Medical Center, 

New York, NY; 3 Ng Medical Group, New York, NY 

Introduction: Despite the proven efficacy of current antiviral 

therapy for chronic hepatitis B (CHB), up to 20% of patients on 

long term oral antiviral treatment who have complete HBV suppression 

have persistently elevated serum alanine transferase 

(ALT) levels. It has been previously shown that non-alcoholic 

fatty liver disease (NAFLD) is a likely culprit. This phenomenon 

will contribute to an increasingly large cohort of patients who 

will develop NAFLD-related cirrhosis, decompensated liver disease, 

and hepatocellular carcinoma. Asians may be especially 

affected given endemic CHB and significant prevalence of metabolic 

syndrome and NAFLD despite lower body mass index 

and rate of obesity compared to other ethnicities. Methods: 

Consecutive Chinese American patients from a single non-transplant 

urban referral center (Mount Sinai Beth Israel Medical 

Center, New York City) who underwent biopsies for CHB 

between January 1, 2008 and Dec 31, 2012 were included in 

this retrospective study. The biopsy specimens were reviewed, 

regraded and restaged for NAFLD according to the scoring 

system designed and validated by Kleiner, Brunt and the 

Pathology Committee of the NASH Clinical Research Network. 

Concomitant HBV grading and staging were also evaluated 

using Scheuer’s classification for chronic hepatitis. Corresponding 

clinical data was gathered. The patients were divided into 

those with NAFLD (+) and without NAFLD (-) on pathology. 

Pathology and clinical features were compared between the 

two groups using the Wilcoxon rank-sum test. Results: Of the 

148 patients, 41 were NAFLD+: 38 with steatosis, 18 with 

steatohepatitis, 13 with steatofibrosis. Two of the patients with 

steatofibrosis had no NAFLD activity but had significant fibrosis 

attributable to NASH. There is a significantly higher combined 

necroinflammatory activity (P=.00001) and overall combined 

staging (P =.0015) in patients in the NAFLD+ group as compared 

to the NAFLD- group. Clinically, alanine transaminase 

(ALT, P=0.0084), body mass index (BMI, P


2216 

The dual and opposite role of the TM6SF2-rs58542926 

Variant in Protecting against Cardiovascular Disease 

and Conferring Risk for Non-alcoholic fatty liver: A 

meta-analysis 

Carlos J. Pirola 2 , Silvia Sookoian 1 ; 1 Clinical and Molecular Hepatology, 

IDIM-CONICET, Ciudad Autonoma de Buenos Aires, 

Argentina; 2 Molecular Genetics and Biology of Complex Diseases, 

IDIM CONICET, Buenos Aires, Argentina 

Background: The nonsynonymous p.Glu167Lys (rs58542926 

C/T, E167K) variant, located in TM6SF2 (transmembrane 6 

superfamily member 2) gene, was associated not only with 

blood lipid levels, including serum total cholesterol (TC), 

low-density lipoprotein cholesterol (LDL-C) and triglycerides 

(TG), but also myocardial infarction risk and NAFLD susceptibility. 

This variant, which has a relatively low frequency not 

only in Caucasian population (0.09) but globally (0.07), presents 

a clinical paradox, as the C (Glu167) allele was shown 

to be associated with increased CVD risk, while the T allele 

(Lys167) was associated with NAFLD. Objective: to examine 

the evidence provided in the available literature to estimate the 

strength of the effect of rs58542926 on both circulating lipid 

traits and NAFLD across different populations. In addition, we 

assessed whether associations are consistent across studies 

in magnitude and direction for all explored traits. Methods/ 

design: We performed a systematic review by a meta-analysis; 

literature searches identified eleven studies. Results: The 

rs58542926 exerts a significant role in modulating lipid traits, 

including TC, LDL-C, TG, and NAFLD. However, this influence 

on lipids and NAFLD is opposite between genotypes in the 

dominant model of inheritance. Pooled estimates of random 

effects in 193,931 individuals showed that, compared with 

carriers of the minor K allele (EK+KK individuals), subjects 

homozygous for the ancestral C allele (EE genotype) are at risk 

of having higher levels of TC, LDL-C and TG; the differences 

in mean±SE (mg/dL) are 7.4±2.3, 3.7±0.9 and 9.4±2.1, 

respectively. The rs58542926 was not associated with HDL-C 

in a large sample (n =91,937). Of note, the EE genotype is 

associated with a relatively large effect on blood levels of TC 

(about 2-3% increase on an upper limit of 200 mg/dL), LDL-C 

(about 1-2% increase on an upper limit of 150 mg/dL), and 

TG (about 6.4% increase on an upper limit of 150 mg/dL) for 

a single variant influencing a polygenic trait. In contrast, homozygous 

EE subjects appear to be protected against NAFLD 

(OR 0.469, 95% CI 0.300-0.734, p = 0.0009, n = 3273), 

while carriers of the K allele show about~ 2.2% higher lipid 

fat content when compared with homozygous EE (n = 3,413), 

and have 2.13-fold higher risk of developing NAFLD. Conclusion: 

The rs58542926 appears to be an important modifier of 

blood lipid traits in different populations. As a challenge for 

the personalized medicine, the “major” C-allele, which has a 

frequency as high as 93%, is associated with CVD risk, while 

the “minor”-low frequency T allele confers risk for NAFLD; in 

turn, CVD and NAFLD are strongly related outcomes. 

Disclosures: 

The following authors have nothing to disclose: Carlos J. Pirola, Silvia Sookoian 

2217 

Mitochondrial Mutator-Phenotype May Be Related 

to Pathogenesis of Nonalcoholic Fatty Liver Disease: 

Insights from Deep Sequencing of Liver Mitochondrial 

Genomes 

Carlos J. Pirola 1 , Romina Scian 1,2 , Cristian O. Rohr 3 , Hernán 

Dopazo 3 , Gustavo O. Castaño 4 , Silvia Sookoian 2 ; 1 Molecular 

Genetics and Biology of Complex Diseases, IDIM-CONICET, 

Buenos Aires, Argentina; 2 Clinical and Molecular Hepatology, 

IDIM-CONICET, Buenos Aires, Argentina; 3 Biomedical Genomics 

and Evolution Laboratory, CONICET, Buenos Aires, Argentina; 

4 Medicine and Surgery Department, Liver Unit, Hospital Abel Zubizarreta, 

Buenos Aires, Argentina 

Background: Mitochondrial (mt) dysfunction is involved in the 

development of NAFLD; normal activity of mitochondria critically 

determines fatty acid beta-oxidation, OXPHOS and insulin 

signaling. In addition, liver mitochondrial biogenesis is reduced 

in NAFLD. The mt-genome is highly polymorphic and variants 

in mtDNA affect mt function. A small proportion of mtDNA 

belongs to the control region involved in mtDNA duplication 

(D-loop). To understand the clinical implications of mtDNA-variation 

in the pathogenesis of NAFLD, we sequenced whole liver 

mtDNA-genomes of 28 individuals, including patients with 

NAFLD (n = 20) and age and sex-matched controls (n = 8). In 

addition, we sequenced the entire nuclear POLG and POLG2 

genes, which are involved in mtDNA-replication. Methods: 

Liver mtDNA was first amplified by long-range PCR; deep next 

generation sequencing was further performed. Results: We 

achieved an average read depth >800 per individual; mtDNA 

sequencing revealed 689 variants, 525 (76%) of them were 

observed in NAFLD showing an enrichment of 1.28-fold mutation 

fraction compared to controls (p = 0.0056). Ten of 16 

base positions containing heteroplasmic variants were highly 

polymorphic (>0.1) and were observed in NAFLD. The mutation 

fraction of liver mtDNA-genomes in controls compared with 

that in patients with simple steatosis (NAFL) shows no significant 

differences. Remarkably, patients with NASH compared 

with controls harbored a significantly higher number of mutations 

in mitochondrially encoded ATP synthase 6 (p = 0.033), 

cytochrome b (MT-CYB) (p = 0.011), and members of the 

NADH-dehydrogenase complex (p = 4.5 E -5 ). The comparison 

of liver mtDNA-diversity among patients with NAFL and NASH 

showed that the disease severity was associated with increased 

number of variants in the NADH-complex (p = 7.4 E -3 ). For variants 

predicted to be deleterious, an allelic association analysis 

was further conducted; we found a nonsynonymous variant in 

MT-CYB (m.15326) and two mutations in D-loop (m.146 and 

m.16298) that were significantly associated with the disease 

severity. Overall, mutations located in the NADH-complex were 

significantly associated with liver-related phenotypes and arterial 

hypertension. The missense p.Gln1236His variant in POLG 

was associated with liver mtDNA-copy number (p = 0.01) and 

the POLG2- rs7223078 was associated with the degree of histological 

steatosis (p = 0.036). Conclusion: The burden of mutations 

in liver mt-genomes may contribute to the pathogenesis 

of NAFLD and metabolic syndrome-associated comorbidities 

explaining part of the “missing heritability”. NASH development 

may be associated with an OXPHOS-negative phenotype. 

Disclosures: 

The following authors have nothing to disclose: Carlos J. Pirola, Romina Scian, 

Cristian O. Rohr, Hernán Dopazo, Gustavo O. Castaño, Silvia Sookoian


2218 

Reverse correlation with FXR expression of peripheral 

mononuclear cell and NPC1L1 expression of terminal 

ileum in non-alcoholic fatty liver disease. 

Sang Bong Ahn 1 , Dae Won Jun 2 , Yong Kyun Cho 3 , Eun Chul Jang 4 , 

Seung Min Lee 5 , Kiseok Jang 6 ; 1 Department of Internal Medicine, 

Eulji University School of Medicine, Seoul, Korea (the Republic of); 

2 Department of Internal Medicine, Hanyang University College of 


Medicine, Kangbuk Samsung Hospital, Sungkyunkwan University, 

School of Medicine, Seoul, Korea (the Republic of); 4 Department 

of Occupational and Environmental Medicine, Soonchunhyang 

University Cheonan Hospital, Cheonan, Korea (the Republic of); 

5 Department of Food and Nutrition, Sungshin Women’s University, 


and Pathology, Hanyang University School of Medicine, Seoul, 


Background: Niemann-Pick C1-Like 1 (NPC1L1) functions as 

sterol transporter to mediate intestinal cholesterol absorption. 

Although the role of Liver X receptor (LXR) and Farnesoid X 

receptor (FXR) in hepatic steatosis is well known, its correlation 

with LXR/FXR and intestinal cholesterol circulation has not been 

thoroughly studied. We investigated the associations among 

LXR, FXR and NPC1L1 as well as its correlation with intestinal 

cholesterol circulation in patients with non-alcoholic fatty liver 

disease (NAFLD). Methods: We evaluated clinical characteristics 

from 25 NAFLD and 28 control patients. We conducted 

upper endoscopy, colonoscopy and non-enhanced CT. Fatty 

liver diagnosed with decreased liver/spleen HU. We calculated 

the expression level of LXR and FXR in peripheral mononuclear 

cell by qPCR. Immunohistochemical analysis (ABCA1, 

ABCG5/8, NPC1L1, SREBP, LXR, FXR, CD36) was carried 

out on tissue samples of duodenum and ileum from NAFLD 

patients. Results: The expression of LXR (p=0.01) and FXR 

(p=0.03) in mononuclear cells increased in NAFLD group compared 

to the control group. Expression of LXR, FXR, NPC1L1, 

ABCA1, ABCG5/8, and SREBP in ileum was decreased in 

fatty liver disease. The expression of LXR in mononuclear cell 

showed a negative correlation with the expression of LXR, 

FXR, ABCA1, ABCG8, SREBP, NPC1L1 in ileum. Expression 

of NPC1L1 (p=0.15), FXR (p=0.30) in ileum was decreased 

in patients with hypercholesterolemia. Expression of NPC1L1 

(p=0.03), LXR (duodenum) and LXR (ileum) was decreased as 

triglyceride increased. Conclusion: We can predict the expression 

of NPC1L1 in terminal ileum by measuring the expression 

of LXR and FXR in peripheral mononuclear cell. Keywords: Liver 

X receptor, Farnesoid X receptor, NPC1L1, non-alcoholic fatty 

liver disease 

Disclosures: 

The following authors have nothing to disclose: Sang Bong Ahn, Dae Won Jun, 

Yong Kyun Cho, Eun Chul Jang, Seung Min Lee, Kiseok Jang 

2219 

Association of Nonalcoholic Fatty Liver Damage and 

the Degree of Nocturnal Hypoxia in Obstructive Sleep 

Apnea 

Erol Cakmak 1 , Faysal Duksal 2 , Engin Altinkaya 3 , Fettah Acibucu 5 , 

Omer T. Dogan 4 , Ozlem Yonem 1 , Abdulkerim Yilmaz 1 ; 1 Department 

of Gastroenterology, Cumhuriyet University School of Medicine, 

Sivas, Turkey; 2 Department of Chest Diseases, Numune 

Hospital, Sivas, Turkey; 3 Department of Gastroenterology, Kayseri 

Training and Research Hospital, Kayseri, Turkey; 4 Department of 

Chest Diseases, Cumhuriyet University School of Medicine, Sivas, 

Turkey; 5 Department of Endocrinology, Numune Hospital, Sivas, 

Turkey 

Background: Nonalcoholic fatty liver disease (NAFLD) has a 

large spectrum of findings ranging from a simple fatty liver to 

advanced fibrosis, cirrhosis and hepatocellular cancer. Obstructive 

sleep apnea (OSA) can lead to the development and progression 

of NAFLD because of repeated nocturnal hypoxia. 

The aim of this study was to evaluate the effect of the nocturnal 

hypoxia severity on hepatic steatosis. Methods: The parameters 

evaluated in this retrospective study in 137 subjects were body 

mass index, biochemical tests, degree of steatosis according 

to the findings of the liver ultrasound, and polysomnographic 

parameters demonstrating the severity of OSA (apnea-hypopnea 

index [AHI], oxygen desaturation index [ODI], mean 

nocturnal SpO2, lowest desaturation value [LaSO2]). According 

to the AHI score, patients with sleep apnea with a score of 

30 were categorized as control, mild, 

intermediate, and severe, respectively. Results: The total number 

of subjects included in this study was 137 (76 females and 

61 males) with a mean age of 56±10 years. Among the 118 

patients who were diagnosed with OSA, 19 had mild OSA, 

39 had intermediate OSA, 60 had severe OSA, and there 

were 19 cases in the control group. AHI and ODI values were 

significantly higher in the intermediate NAFLD (p


2220 

Transient Elastography is Feasible with High Success 

Rate for Evaluation of Non-Alcoholic Fatty Liver Disease 

(NAFLD) in a Multicenter Setting 

Raj Vuppalanchi 1 , Mohammad S. Siddiqui 2 , Erin K. Hallinan 

3 , Manal F. Abdelmalek 4 , Brent A. Neuschwander-Tetri 5 , 

Rohit Loomba 6 , Kris V. Kowdley 7 , Norah Terrault 8 , Arthur J. 

McCullough 9 , James Tonascia 3 , Elizabeth M. Brunt 10 , David E. 

Kleiner 11 , Edward Doo 12 , Naga P. Chalasani 1 , Arun J. Sanyal 2 ; 

1 Indiana University, Indianapolis, IN; 2 Virginia Commonwealth, 

Richmond, VA; 3 Johns Hopkins Bloomberg School of Public Health, 

Baltimore, MD; 4 Duke, Durham, NC; 5 St.Louis University, St.Louis, 

MO; 6 University of California at San Diego, San Diego, CA; 

7 Swedish Medical Center, Seattle, WA; 8 University of California at 

San Francisco, San Francisco, CA; 9 Cleveland Clinic, Cleveland, 

OH; 10 Washington University, St.Louis, MO; 11 NCI, Bethesda, 

MD; 12 NIH, Bethesda, MD 

Background: Vibration-controlled transient elastography using 

FibroScan to obtain the liver stiffness measurement (LSM) and 

controlled attenuation parameter (CAP) is a promising non-invasive 

tool for assessment of hepatic fibrosis and steatosis 

respectively. However, prior studies using older versions of 

FibroScan reported high failure rates in nonalcoholic fatty liver 

disease (NAFLD). Aim: The aim of the current study was to 

examine the feasibility and success of the FibroScan 502 Touch 

in patients with NAFLD who were participating the NAFLD 

Database Study conducted by the NASH Clinical Research 

Network (NASH CRN). Methods: A total of 511 NAFLD 

patients across eight clinical centers were eligible and provided 

informed consent. The FibroScan 502 Touch was used in 

all patients with either the medium (M+) or large (XL+) probes. 

Both LSM and CAP were measured after an overnight fast 

using a standardized protocol. The scanning was performed 

twice (FS1 and FS2) during the same study visit. “Failure” was 

defined as the inability to obtain a valid exam whereas “unreliable” 

was defined as those exams with 30%. Results: The cohort 

included 65% women with a mean (± SD) age of 56 (± 12) 

years, BMI 33.6 (± 6.8) kg/m 2 and waist circumference 107 

(± 14) cm. Based on the recommendations from the automatic 

probe selection tool, XL+ probe was required for the study in 

57% of patients. Five patients (1.0%) refused the procedure 

after providing informed consent and five patients (1.0%) had 

a skin-to-capsule distance greater than 3.5 cm and the exam 

could not be completed. Among remaining 501 patients, the 

results were unreliable in 8 (1.6%). Using logistic regression 

analysis, only female gender [OR: 0.29, 95%CI: 0.10, 0.83, 

p=0.02) and waist circumference [OR: 1.04 per cm, 95%CI: 

1.01, 1.08, p=0.004) were associated with a failed or unreliable 

exam. There was excellent agreement between FS1 and 

FS2 for LSM (r=0.96, 95% CI: 0.95, 0.97) and CAPs (r=0.81, 

95% CI: 0.77, 0.84). In a subset of patients who had a liver 

biopsy within 12 months of the FibroScan (n=74), log-linear 

regression analysis showed statistically significant relationships 

between the geometric mean (GM) LSM and advanced (stage 

3 or 4) vs. no or less advanced fibrosis [GM ratio: 2.04 (95% 

CI: 1.60, 2.60), p


Disclosures: 

The following authors have nothing to disclose: Claudia P. Oliveira, José Tadeu 

Stefano, Roberto M. Ribeiro, Sebastião M. Duarte, Livia Rodrigues, Priscila B. 

Campos, Fernando G. Costa, Daniel F. Mazo, Flair J. Carrilho, Ester C. Sabino 

2222 

Outcome of bariatric surgery, in highly selected morbid 

obese with compensated cirrhosis 

Guillaume Lassailly, Robert Caiazzo, Charlotte Vanveuren, 

gnemmi vivianne, Alexandre Louvet, Emmanuelle Leteurtre, Florent 

Artru, Sebastien Dharancy, Valerie Canva, Francois Pattou, 

Philippe Mathurin; CHRU lille, Lille, France 

Introduction: Morbid obesity contributes to liver function impairment 

in cirrhotic patients and especially in NASH. Weight loss 

strategies could improve liver function, but bariatric surgery 

is not considered due to an increased risk of mortality after 

surgery. However its benefit and feasibility in highly selected 

patients is still unclear. The aim of this study was to describe 

the outcome of highly selected cirrhotic patients after bariatric 

surgery. Methods: Among the 2024 morbid obese patients of 

the Lille Bariatric prospective cohort (1994-2015), 28 compensated 

cirrhotic patients underwent bariatric surgery. When cirrhosis 

was identified before surgery, each case was discussed 

to validate the procedure. Only Child-Pugh A5 patients without 

portal hypertension (without varice and hepatic vein gradient 


Disclosures: 

The following authors have nothing to disclose: Leonardo A. Martinez Rodriguez, 

Aldo Torre, David Kershenobich 

2224 

The association of ALT levels with myocardial perfusion 

imaging and cardiovascular morbidity 

David Yardeni 2 , Ohad Etzion 1,3 , Ronen Toledano 4 , Victor 

Novack 4 , Aryeh Shalev 5 , Arik Wolak 5 , Yaron Rotman 1 ; 1 Liver Disease 

Branch, NIH, Rockville, MD; 2 Internal Medicine Division, 

Soroka University Medical Center, Beer-Sheva, Israel; 3 Gastroenterology 

and Liver disease, Soroka University Medical Center, 

beer-Sheva, Israel; 4 Clinical Research Center, soroka University 

Medical Center, Beer-Sheva, Israel; 5 Department of Cardiology, 

Soroka University Medical Center, beer-Sheva, Israel 

Background and aims: There is a growing body of evidence 

suggesting that non-alcoholic fatty liver disease (NAFLD) is 

associated with an independent increased risk of cardiovascular 

disease (CVD) beyond that is conferred by the metabolic 

syndrome. We utilized a large cohort of patients undergoing 

myocardial perfusion imaging (MPI) with single photon emission 

computed tomography (SPECT) to determine the association 

between alanine aminotransferase (ALT) as a surrogate 

marker for NAFLD,and the presence of myocardial ischemia as 

well as the effects of ALT on mortality in patients with and without 

ischemia. Methods: We retrospectively assessed SPECT-MPI 

and laboratory test results, prescriptions and clinical diagnoses, 

extracted from the electronic medical records of all individuals 

who underwent SPECT MPI at Soroka University Medical 

Center between 1997 and 2008. We excluded patients with 

viral hepatitis, positive autoimmune markers, ALT values 340 u/L, and absent liver tests within 1 year prior or 

following SPECT MPI. Cases with elevated ALT were classified 

as presumed NAFLD. The primary outcome was abnormal myocardial 

perfusion, defined as >1% perfusion defect. Secondary 

outcomes were a composite cardiac outcome of cardiac death 

or acute myocardial infarction 3 and 5 years following SPECT- 

MPI, and all-cause mortality Results: Of 26,028 patients who 

underwent SPECT-MPI, 11,582 met inclusion criteria and were 

included in the final analysis. 1,704 (14.7%) patients had elevated 

ALT within a year of SPECT-MPI. Patients with abnormal 

ALT were younger and included significantly more males and 

smokers (p < 0.001 for all). Dyslipidemia, diabetes mellitus, 

obesity and congestive heart failure were also more common 

in the elevated ALT group. SPECT-MPI results did not differ 

between subjects with elevated ALT and controls. Elevated ALT 

was associated with increased risk for the composite outcome 

of cardiac death or acute myocardial infarction at 3-year follow-up, 

HR, 1.46 (95% CI 1.07–1.99) and in all cause mortality 

(HR. 1.69, CI (1.28–2.23) but only in patients with normal 

SPECT-MPI. At 5-years follow-up ALT levels were not associated 

with the composite outcome or all-cause mortality. Conclusions:. 

The long-term mortality of patients with abnormal SPECT-MPI is 

not modulated by ALT, likely reflecting an already high risk and 

established advanced atherosclerotic process. On the other 

hand, patients with normal SPECT-MPI are at increased risk for 

a future cardiac event if they have an elevated ALT level, suggesting 

an important role for NAFLD in earlier stages of CVD. 

Our results highlight the complex interaction between NAFLD 

and ischemic heart disease. 

Disclosures: 

The following authors have nothing to disclose: David Yardeni, Ohad Etzion, 

Ronen Toledano, Victor Novack, Aryeh Shalev, Arik Wolak, Yaron Rotman 

2225 

Non-Alcoholic Fatty Liver Disease (NALFD) and Hepatocellular 

Carcinoma (HCC), clinical characteristics and 

outcome: A single centre experience. 

Sum Team Lo, David W. Orr, Adam Barlett, Edward J. Gane; New 

Zealand Liver Transplant Unit, Auckland, New Zealand 

Aims: To compare patient survival between NAFLD-related 

HCC and chronic viral hepatitis-related HCC and to examine 

the risk factors and characteristics of NAFLD patients with 

HCC. Methods: A retrospective analysis was undertaken at 

a national hepatocellular carcinoma (HCC) multidisciplinary 

service of all cases of NALFD-related HCCs and matched cases 

with chronic viral hepatitis-related HCCs diagnosed between 

2001 and 2014. Results: A total of 93 patients with NAFLD- 

HCC were diagnosed. These were compared to case-matched 

patients with chronic viral hepatitis; HBV or HCV-related HCC 

(4:1; n=366). The mean age in NAFLD-HCC was 70.8 and 

viral-HCC 58.1 years respectively (p


is a common finding which can be associated with chronic 

liver disease but referral practices vary. Aim: to investigate 

referral practices, prevalance of liver disease and three, five 

and 10 year mortality of minimally deranged ALT. Methods All 

LFT results requested by primary care in Newcastle between 

1 st January and 31 st December 2003 were collected. Patients 

with at least one ALT in the range 41-90IU/L were flagged 

and the patients subsequently referred to Gastroenterology or 

Hepatology were identified and case notes reviewed. Mortality 

was investigated by matching ALT41-90 patients by age, 

gender and GP practice to patients with normal range ALT. 

Up to date mortality data was collected and three, five and 

10 year mortality rates for both groups were calculated and 

compared using chi-square tests. Results In 2003, 56,131 LFTs 

were checked by primary care in Newcastle. Of these, 5,489 

unique patients (9.8%) had an ALT in the range 41-90, while in 

986 (1.8%) ALT was >90. Of the ALT41-90 patients, only 331 

(6%) were referred to Gastroenterology or Hepatology within a 

year. Casenotes were available for review in 309 (93%). 179 

(58%) had evidence of significant liver disease and 22 (7%) 

had established cirrhosis. Mortality: 1,195 (21.7%) ALT41- 

90 patients were matched to normal ALT controls. Mean ALT 

was 55.6 (SD 15.9) and 22.1 (SD 7.7) in the ALT41-90 and 

normal groups respectively. Mean age was 50.2 (SD 14.1) in 

both groups. There was no statistically significant difference 

in the three, five or 10 year mortality between the ALT41-90 

patients and the normal ALT patients (see table). Conclusions 

Minimally deranged ALT is associated with a high risk of liver 

pathology in asymptomatic patients but many of these patients 

are not referred to a specialist. We did not find that a minimally 

deranged ALT was associated with significantly higher 

mortality rates at three, five or 10 years but it is possible that 

this would be observed after longer follow-up. It would be valuable 

to study the relative benefits of other methods of screening 

for asymptomatic liver disease, in particular the combination of 

aspartate aminotransferase (AST) with ALT in the AST:ALT ratio. 

key pathological processes in NAFLD. We sought to determine 

whether changes in VLDL profiles predict NAFLD disease progression. 

Methods: We evaluated VLDL profiles of127 patients 

from a single center NAFLD registry, and examined VLDL size 

and subclass particle concentrations in relation with biopsy-determined 

NASH and liver fibrosis. Results: NASH was associated 

with larger VLDL size, but not higher concentration. In 

a multivariate model, every nm increase in mean VLDL size 

was associated with 2% increase in the relative risk of NASH 

(95% CI 1.01–1.03, p = 0.002). A decrease in small VLDL 

particle concentration was associated with more advanced 

liver fibrosis. Every nmol/L decrease in the concentration of 

small VLDL particles was associated with 2% increase (95% CI 

1.003–1.04, p = 0.03) in the relative risk of stage 2 or above 

fibrosis. Mean VLDL size and small VLDL particle concentration 

alone performed equally well as cytokeratin-18 (CK18) and 

NAFLD fibrosis score (NFS) in predicting both NASH and significant 

liver fibrosis (Figure). The incorporation of VLDL measurements 

improves the predictive value of existing NASH and 

fibrosis indicators. Conclusions: The development of NASH is 

associated with an increase in mean VLDL size, whereas the 

progression of liver fibrosis is associated with a decrease in 

the concentration of small VLDL particles. Profiles of circulating 

VLDL may be used as biomarkers for NAFLD. 

A. Comparison of CK18, VLDL size and the combination of both 

in predicting NASH (NAS score ≥ 4). B. Comparison of NFS, 

small VLDL concentration and the combination of both in predicting 

significant liver fibrosis (stage 2 or above). 

Mortality of normal ALT vs ALT41-90 

Disclosures: 

The following authors have nothing to disclose: James G. Orr, Richard C. 

Thomas, David Jones, Mark Hudson 

2227 

Steatohepatitis and liver fibrosis are predicted by the 

size and subclass concentration of very low density lipoprotein 

in nonalcoholic fatty liver disease 

Zhenghui G. Jiang 1 , Elliot B. Tapper 1 , Margery A. Connelly 2 , Carolina 

F. Pimentel 1 , Linda Feldbrügge 1 , Misung Kim 3 , Sarah Krawczyk 

3 , Nezam H. Afdhal 1 , Simon C. Robson 1 , Mark A. Herman 3 , 

James Otvos 2 , Kenneth Mukamal 4 , Michelle Lai 1 ; 1 Gastroenterology 

and Hepatology, Beth Israel Deaconess Medical Center, 

Harvard Medical School, Boston, MA; 2 LabCorp, Raleigh, NC; 

3 Endocrinology, Beth Israel Deaconess Medical Center, Harvard 

Medical School, Boston, MA; 4 General Internal Medicine, Beth 

Israel Deaconess Medical Center, Harvard Medical School, Boston, 

MA 

Background: A major challenge in the management of nonalcoholic 

fatty liver disease (NAFLD) is to identify patients at 

risk for disease progression characterized by steatohepatitis 

(NASH) and early liver fibrosis. Very low density lipoprotein 

(VLDL) is produced exclusively from the liver, and influenced by 

Disclosures: 

Margery A. Connelly - Employment: LabCorp 









Mark A. Herman - Speaking and Teaching: Pfizer 

The following authors have nothing to disclose: Zhenghui G. Jiang, Elliot B. 

Tapper, Carolina F. Pimentel, Linda Feldbrügge, Misung Kim, Sarah Krawczyk, 

James Otvos, Kenneth Mukamal, Michelle Lai


2228 

Relationships between very low density lipoprotein 

profiles and pathological processes in nonacoholic fatty 

liver disease: insights from the Cardiovascular Health 

Study 

Zhenghui G. Jiang 1 , Ian H. de Boer 2 , Rachel H. Mackey 3 , Majken 

K. Jensen 4 , Michelle Lai 1 , Russel Tracy 3 , Simon C. Robson 1 , Lewis 

Kuller 5 , Kenneth Mukamal 6 ; 1 Gastroenterology and Hepatology, 

Beth Israel Deaconess Medical Center, Harvard Medical School, 

Boston, MA; 2 Nephrology, University of Washington, Seattle, WA; 

3 Pathology, University of Vermont College of Medicine, Burlington, 

VT; 4 Nutrition, Harvard School of Public Health, Boston, MA; 

5 Epidemiology, University of Pittsburgh Graduate School of Public 

Health, Pittsburgh, PA; 6 General Internal Medicine, Beth Israel 

Deaconess Medical Center, Harvard Medical School, Boston, MA 

Background: Nonalcoholic fatty liver disease (NALFD) significantly 

impacts the production of very low density lipoprotein 

(VLDL) from the liver. As a result circulating VLDL profile 

may serve as a marker for NAFLD disease severity. Herein, 

we aim to determine the impact on VLDL profiles from three 

key pathological processes in NAFLD: insulin resistance, liver 

synthetic function and inflammation. Methods: We examined 

cross-sectional associations of markers for insulin sensitivity, 

inflammation, and liver synthetic function with VLDL particle 

(VLDL-P) subclass concentration and size among 1,850 participants 

in the Cardiovascular Health Study. Results: Insulin resistance 

(lower Matsuda index and 1/HOMA-IR) was associated 

with larger mean VLDL size and higher concentrations of large 

VLDL-P. Markers of liver synthetic function (albumin, fibrinogen 

and factor VII) were positively associated with concentrations 

of total and subclass VLDL-P. CRP and IL-6, markers for inflammation, 

demonstrated a nonlinear relationship with both total 

VLDL-P concentration and VLDL size. When mutually adjusted, 

one standard deviation (SD) increment in Matsuda index and 

CRP were associated with -4.9 nmol/L (-8.2 - -1.5, p = 0.005) 

and -6.3 nmol/L (-11.0 - -1.6, p = 0.009) lower VLDL-P concentration 

respectively. In contrast, one-SD increment in factor VII 

was associated with 16.9 nmol/L (12.6 – 21.2, p < 0.001) 

higher VLDL-P concentration. In addition, a one-SD increment in 

the Matsuda index was associated with -1.1 nm (-2.0 - -0.3, p 

= 0.006) smaller mean VLDL size, whereas CRP and factor VII 

were not associated with VLDL size. Conclusion: Insulin resistance 

is associated with higher VLDL-P concentration and larger 

mean VLDL size, whereas both inflammation and impaired liver 

synthetic function are associated with lower concentrations of 

VLDL-P, but have no relationship with VLDL size. These differential 

impacts on VLDL profile set the foundation for the utility of 

VLDL in disease staging of NAFLD. 

Adjusted regression coefficients for VLDL profiles from standardized 

Matsuda index, CRP and factor VII 

Regression models calculated using the Z-scores of three primary 

exposures (Mastuda index, CRP, factor VII) in units of standard 

deviation (SD), and covariates including age, gender, ethnicity, 

BMI, alcohol, smoking history and the use of lipid lowering medications. 

The following authors have nothing to disclose: Zhenghui G. Jiang, Ian H. de 

Boer, Rachel H. Mackey, Majken K. Jensen, Michelle Lai, Russel Tracy, Lewis 

Kuller, Kenneth Mukamal 

2229 

WITHDRAWN 

2230 

The development of hepatocellular carcinoma in Japanese 

patients with biopsy-proven nonalcoholic fatty liver 

disease : the relation between carriage of the PNPLA3 

rs738409 C >G polymorphism and hepatocarcinogenesis 

Yuya Seko 1 , Yoshio Sumida 1 , Saiyu Tanaka 2 , Hiroyoshi Taketani 1 , 

Kazuyuki Kanemasa 2 , Hiroshi Ishiba 1 , Akira Okajima 1 , Tasuku 

Hara 1 , Kanji Yamaguchi 1 , Michihisa Moriguchi 1 , Kohichiroh 

Yasui 1 , Hironori Mitsuyoshi 1 , Yoshito Itoh 1 ; 1 Kyoto Prefectural University 

of Medicine, Kyoto, Japan; 2 Center for Digestive and Liver 

Diseases, Nara City Hospital, Nara, Japan 

BACKGROUND: Some patients with nonalcoholic fatty liver disease 

(NAFLD) develop hepatocellular carcinoma (HCC). Carriage 

of the PNPLA3 rs738409 (encoding the I148M variant) 

has been reported to be associated with advanced fibrosis 

and HCC related to NAFLD. Our aim was to determine the 

incidence of and risk factors including PNPLA3 rs738409 polymorphism 

for HCC in Japanese patients with biopsy-proven 

NAFLD. METHODS: This retrospective cohort study analyzed 

hepatocarcinogenesis in 341 patients with biopsy-proven 

NAFLD. PNPLA3 rs738409 genotype was determined by allelic 

discrimination in 133 patients. RESULTS: Of 341 patients, 205 

(60.1%) were diagnosed with nonalcoholic steatohepatitis. The 

PNPLA3 rs738409 genotype frequencies were CC 0.14, CG 

0.46, GG 0.40. During a median follow-up period of 5.0 

years, 9 patients (2.6%) developed HCC. The cumulative rate 

of HCC was 1.5% at the end of the 5th year and 7.1% at the 

end of the 10th year. Multivariate analysis identified PNPLA3 

genotype (GG; hazard ratio [HR] 14.2, p=0.035), and platelet 

count (^15x10/μl; HR 11.6, p=0.036) as predictors for the 

development of HCC. PNPLA3 genotype was analysed in 9 

patients with HCC, 7 were PNPLA3 GG and 2 were CG. In 

patients with GG allele, the cumulative rate of HCC was 4.9% 

at the end of the 5th year and 33.5% at the end of the 10th 

year. The incidence of HCC was significantly higher in patients 

with PNPLA3 GG allele than in those with CC and CG allele 

(p=0.027). CONCLUSIONS: Severe fibrosis was a predictor 

for HCC in NAFLD patients. And PNPLA3 polymorphism genotyping 

was also the risk to the development of HCC. These 

findings might help us HCC surveillance in NAFLD patients. 

Disclosures: 






The following authors have nothing to disclose: Yuya Seko, Yoshio Sumida, Saiyu 

Tanaka, Hiroyoshi Taketani, Kazuyuki Kanemasa, Hiroshi Ishiba, Akira Okajima, 

Tasuku Hara, Kanji Yamaguchi, Michihisa Moriguchi, Kohichiroh Yasui, Hironori 

Mitsuyoshi 

Disclosures: 




Nanopharma, Puretech


2231 

High prevalence of undiagnosed liver cirrhosis and 

advanced fibrosis in type 2 diabetic outpatients 

Francisco Barrera, Juan P. Arab, Carlos E. Benítez, Arnoldo 

Riquelme, Marco Arrese; Department of Gastroenterology, Escuela 

de Medicina, Pontificia Universidad Catolica de Chile, Santiago, 

Chile 

BACKGROUND: Patients with Type 2 Diabetes Mellitus (T2DM) 

are at risk for developing end-stage liver disease due to nonalcoholic 

steatohepatitis (NASH). Non-invasive methods have 

been validated for assessing the severity of nonalcoholic fatty 

liver disease (NAFLD). Data on prevalence of advanced fibrosis 

among T2DM patients evaluated by these methods is scarce. 

AIM: To evaluate prevalence of steatosis, advanced fibrosis 

and cirrhosis by non-invasive methods in T2DM patients. 

METHODS: We conducted a cross-sectional study in 145 consecutive 

T2DM patients (>55 years-old). The presence of cirrhosis 

and advanced fibrosis was evaluated by liver morphology 

assessed by magnetic resonance imaging (MRI) and NAFLD 

fibrosis score (NFS) respectively. Exclusion criteria included 

significant alcohol consumption, viral hepatitis or other liver 

diseases and exposure to hepatotoxic agents. RESULTS: 52.6% 

were women, average age was 60 years (57-64), BMI was 

29.6±4.7 kg/m 2 and diabetes duration was 7.6±6.9 years. 

A high prevalence of liver steatosis (62.4%) and steatosis 

and abnormal ALT (37.6%) was found. The prevalence of 

advanced fibrosis using the NFS was 12.8 and evidence of 

liver cirrhosis on MRI was 5.3%. In a multivariate analysis GGT 

>82 IU/L (P=0.004) and no alcohol intake (P=0.032) were 

independently associated to advanced fibrosis. CONCLUSION: 

A high frequency of undiagnosed advanced fibrosis and cirrhosis 

was observed in otherwise unselected T2DM patients older 

than 55 y/o. These patients are at high risk of developing liver-related 

complications such as portal hypertension and hepatocellular 

carcinoma. Routine screening for liver disease should 

be considered in this population (Grant Support: FONDECYT 

1150327 to M.A.and 1150311 to F.B) . 

Disclosures: 

The following authors have nothing to disclose: Francisco Barrera, Juan P. Arab, 

Carlos E. Benítez, Arnoldo Riquelme, Marco Arrese 

2232 

Discovery of plasma biomarkers for NAFLD using high 

resolution metabolomics 

Miriam B. Vos, Ran Jin, Sophia Banton, Shuzhao Li, Dean P. Jones; 

Emory University, Atlanta, GA 

Purpose: Past attempts to develop diagnostic tests for NAFLD 

using single metabolite biomarkers have failed in part because 

of the heterogeneity of NAFLD. High throughput, high resolution 

metabolomics is rapid and relatively inexpensive, making it a 

promising platform for disease identification and characterization. 

Methods: We performed an exploratory, unbiased metabolomics 

pilot study to develop a biomarker panel for detecting 

NAFLD. Fasting plasma samples of 39 Hispanic-American adolescents 

with either MRS-documented NAFLD (intrahepatic fat > 

5%) or age and BMI matched control individuals (intrahepatic 

fat < 5%) were analyzed using LC-MS high-resolution metabolomics. 

A total of 9,583 metabolite features were yielded 

initially. The top discriminatory and identifiable metabolites 

were selected. Multivariate receiver operating characteristic 

(ROC) curves were generated for individual metabolites and 

combinations of metabolites, respectively. Results: Univariate 

ROC curves >0.80 were observed with 7 of the top metabolites, 

which included metabolites from de novo lipogenesis, 

ketogenic amino acids and hormones. Multivariate ROC analysis 

with cross validation using balanced subsampling demonstrated 

that combinations of the top 7, 10, and 16 selected 

metabolites had higher prediction for NAFLD (AUC>0.9 for all 

panels, Figure 1) compared to single metabolites. Conclusions: 

This pilot “omics”-based biomarker development study shows 

feasibility in using a metabolomics based biomarker panel to 

identify NAFLD with high sensitivity and specificity. Independent 

validation studies are warranted to confirm clinical utility. 

Disclosures: 

The following authors have nothing to disclose: Miriam B. Vos, Ran Jin, Sophia 

Banton, Shuzhao Li, Dean P. Jones 

2233 

Polychlorinated biphenyl exposure in Anniston, Alabama 

is associated with elevated prevalence of liver 

disease 

Heather B. Clair 1 , Keith C. Falkner 1 , Russell A. Prough 2 , Christina 

Pinkston 1 , Guy N. Brock 1 , Marian Pavuk 3 , N. Dutton 3 , Matthew 

C. Cave 1,4 ; 1 Department of Medicine/GI, University of Louisville, 

Louisville, KY; 2 Biochemistry, University of Louisville, Louisville, KY; 

3 CDC, Atlanta, GA; 4 Robley Rex VAMC, Louisville, KY 

Purpose: Elevated serum concentrations of Polychlorinated 

Biphenyls (PCBs) are associated with increased liver injury 

in the general US population. Residents of Anniston, AL are 

highly exposed to PCBs, resulting in a mean serum PCB concentrations 

3-4 times the mean of the general US population. 

An adult cohort of Anniston residents (Anniston Community 

Health Survey – ACHS) was assembled to study health effects 

and PCB exposure. We report elevated liver injury biomarkers 

characteristic of Toxicant-Associated Steatohepatitis (TASH), 

cytokine and adipokine abnormalities. A resampling of this 

population provided longitudinal data related to liver disease, 

metabolic abnormalities and total PCB load, as well as an 

expanded analysis of the dioxin-like PCB congener concentration 

in Anniston residents. Methods: Subjects were recruited 

from residences within the city limits of Anniston, Alabama. 

Serum samples from 738 of the individuals recruited in 2005- 

2007 (ACHS1), and 352 individuals re-sampled in 2014 

(ACHS2) were included in our analysis. In both groups, whole 

(CK18 M65) and caspase-cleaved cytokeratin 18 (CK18 M30) 

were measured using the PEVIVA ELISAs. Serum cytokine/


adipokines were measured using the Luminex cytokine bead 

array. We also analyzed ACHS2 samples for clinical indicators 

of liver injury. Results: ACHS1 was stratified into three 

groups based on CK18: no liver disease (None, M30


0.838±0.035 and 0.816±0.036 respectively, FIB4 (cut-off 

1.4, Se=77%, Sp=79.8%, and DA=78.6%) and FM-NAFLD 

(cut-off 0.6; Se=68.9%, Sp=85.7%, and DA=78.6%) outperformed 

other models in predicting moderate (F2) hepatic 

fibrosis (P15 e ≤15. Results: 

The sample included 37severe obese patients. The mean age 

was 38.1(SD=9.8) and 51.4% were women. Steatosis was 

observed in 43.2% (16) of them and NASH with and without 

fibrosis in 57% (21). Comparing the 2 groups, patients 

with NASH presented higher levels of ROS [Md 1 

: 11621 

(IIQ:954-30033) versus Md 2 

: 2485 (IIQ:1206-5564) photons, 

p


2238 

Non-alcoholic fatty liver disease: assessment of intestinal 

microbiota and metabolites 

Hannah Da Silva 1,2 , Anastasia Teterina 1 , Bianca M. Arendt 1 , 

Marialena Mouzaki 3 , Sandra E. Fischer 4 , Scott Fung 5 , Johane P. 

Allard 5 ; 1 Gastroenterology, Toronto General Hospital, Toronto, 

ON, Canada; 2 Department of Nutritional Sciences, University of 

Toronto, Toronto, ON, Canada; 3 GI/Hepatology/Nutrition, Hospital 

for Sick Children, Toronto, ON, Canada; 4 Pathology, University 

Health Network, Toronto, ON, Canada; 5 Medicine, Toronto 

General Hospital, Toronto, ON, Canada 

Background: Intestinal microbiota (IM) may contribute to non-alcoholic 

fatty liver disease (NAFLD) through products of bacterial 

metabolism. This study aimed to characterize the bacterial 

products present in serum and stool of NAFLD patients and 

HC and to examine the relationship between these products 

and IM. Methods: This was a prospective, cross-sectional 

study. Fecal and serum metabolite profiles of 39 patients with 

biopsy-proven NAFLD and 28 living liver donors as HC were 

examined using nuclear magnetic resonance spectroscopy. 

Metabolic profiles included 11 fecal and 9 serum bacterial 

products. In a subset of 29 NAFLD and 16 HC, IM composition 

was assessed. Clinical, biochemical, and anthropometric 

data were also collected. Quantitative real-time polymerase 

chain reaction was used to measure total bacterial counts, 

Firmicutes:Bacteroidetes ratio, Bacteroidetes, C. leptum, C. 

coccoides, Bifidobacteria, E. coli and Archaea in stool. Continuous 

variables were non-normal and described as median 

(Q1, Q3). Groups were compared using Wilcoxon rank-sum 

tests. Spearman correlation coefficients were used to examine 

association between IM and bacterial products. Results: 

NAFLD patients were older than HC with higher BMI, liver 

enzymes (AST, ALT), insulin resistance (HOMA-IR), hemoglobin 

A1c, and blood triglycerides (p














NGM 






Dawn M. Torres - Advisory Committees or Review Panels: Genetech; Speaking 

and Teaching: Merck 









Sciences 









The following authors have nothing to disclose: Arthur J. McCullough 

2240 

The effectiveness of SGLT-2 inhibitors as second-line 

treatments for NAFLD patients with type 2 diabetes 

mellitus who failed to incretin based therapy including 

GLP-1 analogues and DPP-4 inhibitors 

Takamasa Ohki 1 , Isogawa Akihiro 2 , Nobuo Toda 1 ; 1 Gastroenterology, 

Mitsui Memorial Hospital, Tokyo, Japan; 2 Diabetes and 

Metabolism, Mitsui Memorial Hospital, Tokyo, Japan 

Background: We already reported that incretin based medicine, 

such as GLP-1 analogues or DPP-4 inhibitors, improved 

glycaemic control and liver inflammation in non-alcoholic fatty 

liver disease (NAFLD) patients with type 2 diabetes mellitus 

(T2DM). However, the effect on ALT normalization was still 

limited. Aims: The aim of this study is to elucidate the effectiveness 

of sodium-glucose co-transporter 2 (SGLT-2) inhibitors 

as second-line treatments for NAFLD patients with T2DM who 

failed to incretin based therapy. Methods: We retrospectively 

enrolled consecutive 130 Japanese NAFLD patients with T2DM 

who were treated with GLP-1 analogues or DPP-4 inhibitors. 

Among them, 70 (53.8%) patients achieved normal ALT level 

and the rest 60 (46.2%) patients did not. We finally obtained 

informed consent from 24 (40.0%) patients out of 60 patients 

and administered SGLT-2 inhibitors in addition to GLP-1 analogues 

or DPP-4 inhibitors. We compared the changes of laboratory 

data including ALT level and body weight at the end of 

follow-up. Results: Thirteen patients were used in combination 

with DPP-4 inhibitors and rest 11 patients were used in combination 

with GLP-1 analogues. The median dosing period was 

320 days. At the end of follow-up, body weight significantly 

decreased (84.8 kg to 81.7 kg, P < 0.01) with amelioration 

of HbA1c level (8.4% to 7.6%, P < 0.01). Serum ALT level 

also significantly decreased (62 IU/l to 49 IU/l, P < 0.01) 

with improvement of FIB-4 index (1.75 to 1.39, P = 0.04). 

Univariate analysis indicated that 3% reduction of body weight 

(OR: 12.0, 95%CI; 1.18-122, P = 0.04) as a factor which 

contributed to normalization of serum ALT level. Conclusions: 

Administration of SGLT-2 inhibitors led not only good glycaemic 

control but also reduction of body weight, normalization of 

ALT level, and alteration of FIB-4 index, even in patients who 

failed to incretin based therapy. 

Disclosures: 

Takamasa Ohki - Speaking and Teaching: Otsuka Pharmaceutical Co. Ltd. 

The following authors have nothing to disclose: Isogawa Akihiro, Nobuo Toda 

2241 

The impact of alcohol consumption for hepatocarcinogenesis 

in Japanese fatty liver disease patients 

Yusuke Kawamura 1,2 , Yasuji Arase 1,3 , Kenji Ikeda 1 , Yushi Sorin 1 , 

Hideo Kunimoto 1 , Hitomi Sezaki 1 , Tetsuya Hosaka 1 , Norio Akuta 1 , 

Masahiro Kobayashi 1 , Satoshi Saitoh 1 , Fumitaka Suzuki 1 , Yoshiyuki 

Suzuki 1 , Mie Inao 2 , Satoshi Mochida 2 , Hiromitsu Kumada 1 ; 

1 Hepatology, Toranomon Hospital, Tokyo, Japan; 2 Gastroenterology 

and Hepatology, Saitama Medical University, Saitama, 

Japan; 3 Health Management Center, Toranomon Hospital, Tokyo, 

Japan 

Background & Aims: The effect of ethanol consumption for 

hepatocarcinogeneis of fatty liver is not clear. The aims of this 

study were to determine the influence of alcohol consumption 

on hepatocarcinogenesis and the risk factors for hepatocellular 

carcinoma (HCC) in a large population of Japanese fatty liver 

patients without viral hepatitis. Methods: This study was retrospective 

cohort study and setting was specialized center for 

hepatology. The study subjects were 9,959 patients with fatty 

liver disease (FLD) without viral hepatitis diagnosed by ultrasonography. 

In this study, the levels of daily ethanol consumption 

divided into following four category;


Disclosures: 

Kenji Ikeda - Speaking and Teaching: Eisai company, Dainippon Sumitomo 

Pharmaceutical company 

Norio Akuta - Patent Held/Filed: SRL. Inc.; Speaking and Teaching: Mitsubishi 

Tanabe Pharma, Janssen Pharmaceutical K.K., Bristol-Myers Squibb 

Fumitaka Suzuki - Speaking and Teaching: BMS 

Yoshiyuki Suzuki - Speaking and Teaching: Bristol-Myers Squibb 



Hiromitsu Kumada - Patent Held/Filed: SRL; Speaking and Teaching: Bristol-Myers 

Squibb,Pharma International, MSD, Janssen Pharma., Glaxosmithkline 

The following authors have nothing to disclose: Yusuke Kawamura, Yasuji 

Arase, Yushi Sorin, Hideo Kunimoto, Hitomi Sezaki, Tetsuya Hosaka, Masahiro 

Kobayashi, Satoshi Saitoh, Mie Inao 

2242 

WITHDRAWN 

2243 

Differences of lifestyle behaviors between non-obese 

and obese non-alcoholic fatty liver disease; beyond visceral 

obesity and metabolic syndrome 

Joo Hee Kwak 1 , Dae Won Jun 1 , Seung Min Lee 2 , Yong Kyun 

Cho 3 , Eun Chul Jang 4 ; 1 Department of Internal Medicine, Hanyang 


2 Department of Food and Nutrition, Sungshin Women’s University, 


Kangbuk Samsung Hospital, Sungkyunkwan University, School of 

Medicine, Seoul, Korea (the Republic of); 4 Department of Occupational 

and Environmental Medicine, Soonchunhyang University 

Cheonan Hospital, Cheonan, Korea (the Republic of) 

Background and Aims: Best way to treat non-alcoholic fatty 

liver disease (NAFLD) is weight reduction. bu However, strategy 

of lifestyle modification in non-obese NAFLD patient is 

unclear. The aim of study was to investigate differences of 

lifestyle behaviors between non-obese and obese non-alcoholic 

fatty liver disease. Methods: This study has been performed 

with 209 patients who wanted to participate in nutrition education 

program because of their abnormal liver enzyme or 

fatty liver. All participants undertook computed tomography. 

NAFLD was diagnosed when liver/spleen Hounsfield Unit (HU) 

was less than 1.1. Five day food record and physical activity 

was measured by two dietitians. Results: Prevalence of NAFLD 

was significantly higher as 66.1% in obese subjects (BMI >25) 

than 42.4% in normal weight (BMI≤25) (p=0.001). Non-obese 

NAFLD group showed higher visceral fat area, aminotransferase 

activity, and fasting glucose compare to normal liver/ 

spleen HU subjects. In univariate analysis, non-obese NAFLD 

group showed higher carbohydrate intake (281 g/day vs. 246 

g/day), and did lower moderate level exercise compare to 

control group (p=0.013). But total calorie intake (1,955 Kcal 

vs. 1,837 Kcal) and walking time/week were not difference. 

In multivariate analysis, the amount of carbohydrate (p=0.018) 

and more than 2 hours of exercise a week (p=0.010) were 

risk factors for NAFLD independent with visceral fat area and 

total calorie intake. Meanwhile fasting glucose (p25). Conclusions: The moderate level exercise of 2 hours a 

week and the amount of carbohydrate intake were independent 

risk factor in non-obese NAFLD patients. 

Disclosures: 

The following authors have nothing to disclose: Joo Hee Kwak, Dae Won Jun, 

Seung Min Lee, Yong Kyun Cho, Eun Chul Jang 

2244 

Subtyping nonalcoholic steatohepatitis for the development 

of effective treatments 

Ibon Martinez-Arranz 1 , Cristina Alonso 1 , David Fernández 2 , 

Rebeca Mayo 1 , Marta Varela 2 , Mazen Noureddin 3 , Maria L. Martinez-Chantar 

2 , Shelly C. Lu 3 , Azucena Castro 1 , Jose M. Mato 2 ; 

1 OWL, Derio, Spain; 2 CIC bioGUNE, Ciberehd, Derio, Spain; 

3 Division of Gastroenterology, Cedars-Sinai Medical Center, Los 

Angeles, CA 

NASH (nonalcoholic steatohepatitis) is a histopathological 

definition that groups together defects in diverse biochemical 

processes that cause hepatic fat accumulation, inflammation 

and necrosis as it were a single disease. The identification 

of the types of mechanisms leading to NASH, together with 

the development of noninvasive biomarkers of these different 

NASH subtypes, is central for the development of precision 

treatments, since the pathways that should be targeted will 

depend upon the specific subtype of NASH. For tissues whose 

primary physiological role is metabolic, such as the liver, 

metabolites are unquestionably the best indicators of organ 

function. Therefore, the detailed analysis of serum metabolome 

may reveal metabolic profiles that may be used as disease 

biomarkers. We previously found that hepatic methionine adenosyltransferase 

(MAT, the enzyme that synthesizes S-adenosylmethionine 

or SAMe) deletion (Mat1a KO) in mice led to the 

spontaneous development of NASH and liver cancer. NASH 

patients often show reduced expression of MAT1A, indicating 

that the Mat1a KO mouse model is relevant to study human 

NASH. Here we have analyzed the serum lipidome (over 400 

different molecular species) in 10-11 month old Mat1a KO 

with histological diagnose of NASH and wild type mice with 

normal liver (twelve animals per group), and selected the top 

fifty serum lipids that more significantly differentiated between 

both genotypes. Then, we observed that this serum biomarker 

panel robustly classified a cohort of 387 patients with biopsy 

proven NAFLD into two well-defined clusters, based on optimum 

average silhouette width, with around 66% of the patients 

showing a MAT1A metabolic subtype (M-subtype). Since there 

is considerable interest in the utility of SAMe to ameliorate 

NAFLD, these results suggest that NAFLD patients with a serum 

M-subtype are best suited to be selected to study the efficacy 

of SAMe treatment. 

Disclosures: 

The following authors have nothing to disclose: Ibon Martinez-Arranz, Cristina 

Alonso, David Fernández, Rebeca Mayo, Marta Varela, Mazen Noureddin, 

Maria L. Martinez-Chantar, Shelly C. Lu, Azucena Castro, Jose M. Mato


2245 

Non-alcoholic fatty liver is not associated with incident 

chronic kidney disease: a large histology based case 

controlled study 

Neeraj Saraf, Narendra S. Choudhary, Sanjiv Saigal, Naveen 

Kumar, Rahul Rai, Dheeraj Gautam, Arvinder Soin; Hepatology, 

Medanta Medicity, Gurgaon, India 

Background: Non-alcoholic steatohepatitis or fibrosis is associated 

with the increased prevalence of impaired kidney function. 

It is not known whether non alcoholic fatty liver (NAFL) 

which is steatosis without inflammation or fibrosis is associated 

with renal impairment as these subjects are not candidates for 

liver biopsy. Materials and methods: The study group included 

all liver donors who underwent donor hepatectomy at our centre 

and had a preoperative liver biopsy for various reasons 

like, high body mass index, dyslipidemia, presence of metabolic 

syndrome or evidence of liver steatosis on imaging. All 

the subjects had negative viral markers. Non-alcoholic fatty 

liver (NAFL) was defined as >5% hepatocytes having steatosis 

and no changes of steatohepatitis and/or fibrosis. Subjects 

with NAFL were compared with subjects with normal liver histology. 

Estimated glomerular filtration rate was calculated with 

Modification of Diet in Renal Disease (MDRD) and CKD-EPI 

method. All subjects had assessment for proteinuria and serum 

creatinine. Results: The study group aged 35±10.3 years and 

mean BMI was 26±3.2 kg/m 2 . One hundred and eighty seven 

adults having NAFL (80 males) were compared to 186 (88 

males) subjects with normal liver histology (controls). Subjects 

with steatosis had significantly higher body mass index 

(26.8±3.5 versus 25.5±3.8 kg/m 2 , P


CVC and PGZ exposures were slightly lower when both drugs 

were co-administered, with a modest decrease in steady-state 

C max 

and AUC 0–tau 

for both drugs, whereas C min 

remained 

unchanged; GMRs for both drugs were ≥0.80 (Table). Effects 

of co-administration on PGZ M-III and M-IV metabolites were 

less pronounced, with 90% CI for systemic exposure ratios 

within the 0.80–1.25 ‘no effect’ range for all 3 PK parameters 

(data not shown). Combination treatment was well tolerated, 

with no serious AEs or AEs leading to discontinuation. All AEs 

were of mild severity and the 2 most commonly reported were 

headache and fatigue. Conclusions: Co-administration of CVC 

and PGZ was well tolerated and resulted in a modest interaction 

that was not considered clinically significant, which suggests 

that dose adjustment is not required when both drugs are 

used in combination. 

(62.1%) male, with a mean age of 57.7 years. 54 (47%) had 

not ultrasonography signals of NAFLD. Between those with 

NAFLD (n=61; 53.1%), 34 (29,6%) had NAFLD grade I, 19 

(16,5%) grade II and 8 (7%) grade III. 26 (22.4%) patients 

had no signals of CAD on their coronary angiography. In 50 

(43.1%), serious injury has found. NAFLD showed correlation 

with CAD in 57 (64.04%) patients (p 25kg/m 2 ), and 15 patients with 

alcoholic (ASH) cirrhosis. Hemostatic status was assessed by 

basal and agonist-induced platelet activation using flow cytometry, 

thrombin generation testing, thromboelastography (TEG), 

a plasma-based clot lysis assay, and an analysis of fibrin pore 

structure by permeation analyses. Results: Basal and agonist-induced 

platelet activation were comparable between patients 

with NAFLD and controls, but agonist-induced platelet activation 

was decreased in patients with cirrhosis. Thrombomodulin-modified 

thrombin generation was comparable between 

patients and controls, although patients with cirrhosis had a 

reduced anticoagulant response to thrombomodulin. TEG test 

results were comparable between controls and non-cirrhotic 

NAFLD patients, but revealed moderate hypocoagulability 

in cirrhosis. Plasma fibrinolytic potential was decreased in 

NAFLD, but accelerated fibrinolysis was observed in ASH cirrhosis. 

Clot permeability was decreased in overweight controls 

and patients with NAFLD. Platelet activity, thrombin generation, 

and clot lysis test results showed a higher variability in 

patients than in controls, suggesting individual patients may 

have a more thrombogenic hemostatic profile. Conclusions: The


combined results of this study show that the overall hemostatic 

status is comparable between patients with NAFLD and controls, 

which contrasts with previously published results in similar 

populations. However, our data show some pro-thrombotic 

features in patients with NAFLD, including hypofibrinolysis and 

a pro-thrombotic structure of the fibrin clot, the latter of which 

appears driven by obesity rather than NAFLD. Our study suggests 

that the role for hyperactive hemostasis in the increased 

risk of thrombosis in patients with NAFLD is probably limited. 

Disclosures: 






The following authors have nothing to disclose: Wilma Potze, Mohammad S. 

Siddiqui, Sherry L. Boyett, Jelle Adelmeijer, Kalyani Daita, Ton Lisman 

2250 

Characteristics and Clinical Outcomes of A Diverse 

Cohort of Patients with Biopsy-Proven Non-Alcoholic 


Benjamin Yip 1 , Brittany E. Yee 2,4 , Ailinh Do 3,4 , Nghia H. 

Nguyen 2,4 , Joseph K. Hoang 4 , Mindie H. Nguyen 4 ; 1 Department 

of Internal Medicine, University of California, Irvine, Orange, CA; 


Diego, CA; 3 School of Medicine, Texas Tech University, El Paso, 

TX; 4 Division of Gastroenterology and Hepatology, Stanford University 

Medical Center, Palo Alto, CA 

PURPOSE: NAFLD is the most common cause of chronic liver 

disease in the U.S. and is associated with increased rates of 

all-cause mortality with known ethnic disparities, though little is 

known about NAFLD in Asian populations. This study aims to 

evaluate presentation and outcomes of patients with NAFLD in 

a cohort of ethnically diverse U.S. cohort. METHODS: We identified 

a cohort of 696 consecutive Asian (n=71) and non-Asian 

(n=625: 41% White, 16% Hispanic, 3% Black, and 30% other) 

patients who underwent liver biopsy showing at least mild steatosis 

(greater than or equal to 5%) who were evaluated at a 

university hospital. Patients with significant alcohol use (> 2 

drinks a day or > 14 drinks a week) were excluded. RESULTS: 

The majority (67%) were female with a median age of 49 

(18-82). The average alcohol intake was 0.29 ± 0.44 drinks 

a day. Concurrent liver diseases included chronic viral hepatitis 

in 12% and other liver disease (including autoimmune, 

iatrogenic, and hereditary liver disease) in 5%. Hypertension 

was common (64%) as well as dyslipidemia (49%) and diabetes 

mellitus (23%). About half underwent liver biopsy during 

another surgical intervention (43% were for weight reduction 

and 10% others) with most of the remaining for work up of 

liver disease (43%). Asians had similar NAFLD activity scores 

(NAS) on liver biopsy as non-Asians (2.08 ± 1.13 vs. 2.00 ± 

1.02, p=0.54) with similar rates of non-alcoholic steatohepatitis 

(NASH, 24% vs. 21%, respectively, p=0.61). However, 

the Asian cohort had higher rates of subsequent cirrhosis (30% 

vs. 17%, p=0.009), HCC (23% vs. 5%, p < 0.0001), and 

orthotopic liver transplantation (13% vs. 4%, p=0.002). On 

multivariate analysis, Asian ethnicity was a significant predictor 

for HCC (OR=3.20, CI 1.31-7.82, p=0.01) but not for 

developing cirrhosis (OR=1.56, CI 0.84-2.91, p=0.16), after 

adjusting for age, gender, daily alcohol use, NAS, the presence 

of NASH, cirrhosis, and other liver diseases. The presence 

of another liver disease was also a significant predictor 

for cirrhosis (OR 3.03, CI 1.83-5.01, p < 0.0001) and HCC 

(OR 13.02, CI 5.49-30.94, p < 0.0001). CONCLUSION: The 

majority of patients with NAFLD in this study underwent incidental 

liver biopsy during surgery. Asian patients with NAFLD 

have significantly higher risk for HCC compared to other ethnicities, 

though they presented with similar NAS and presence 

of NASH, cirrhosis, and secondary liver disease. 

Disclosures: 





The following authors have nothing to disclose: Benjamin Yip, Brittany E. Yee, 

Ailinh Do, Nghia H. Nguyen, Joseph K. Hoang 

2251 

Normal body mass index as a risk factor for osteopenia 

in patients with non-alcoholic fatty liver disease patients 

Maria Farrugia 2 , Martina Gerada 2 , John Bonello 2 , James Mario 

Gauci 2 , Richard Pullicino 1 , Jurgen Gerada 2 ; 1 Radiology, Mater 

Dei Hospital, Msida, Malta; 2 Gastroenterology, Mater Dei Hospital, 

Msida, Malta 

Background/Aim: The effect of non-alcoholic fatty liver disease 

(NAFLD) on bone mineral density (BMD) is poorly understood. 

NAFLD is more prevalent with higher body mass indexes (BMI). 

We aimed to evaluate the effect of different classes of BMI on 

BMD in adult NAFLD patients. Methods: Adult patients diagnosed 

with NAFLD on liver imaging in 2013 were enrolled 

in the study. Patients with concomitant liver pathologies were 

excluded. Patient demographics were obtained from the medical 

notes and the BMI calculated and classified using WHO 

classification (normal (18.5-24.9), overweight (25-29.9) and 

obese (≥30)). BMDs of the femur and lumbar spine were measured 

in all patients by dual energy X-ray absorptiometry. 

Age and gender-matched Z scores and T scores were calculated 

and analysed against the different BMI classes using the 

ANOVA model. Results: 197 NAFLD patients were enrolled in 

the study (178 females, 90.3%; mean age 63.5, range 35-82; 

mean BMI 32.8, range 22-48.6). Obese patients had higher 

BMD T scores of the whole femur than overweight patients and 

normal BMI patients (p 0.001)(Table 1). This was also true 

for the femoral neck (p 0.007) and lumbar spine (p 0.017). 

Likewise, obese patients had higher BMD Z scores of the whole 

femur (p 0.002), femoral neck (p 0.005) and lumbar spine 

(p 0.008) compared to overweight patients and normal BMI 

patients. On subgroup analysis of obese patients, obese class 

3 patients (BMI >40) had higher BMD T scores of the whole 

femur (p 0.002) and lumbar spine (p 0.003) but not of the 

femoral neck (p 0.321) when compared to obese class 2 (BMI 

35-39.9) and obese class 1 (BMI 30-34.9) patients. Conclusions: 

In NAFLD patients, obesity was associated with stronger 

bone mineralisation, demonstrating a linear relationship 

with increasing BMIs. NAFLD patients with normal BMI should 

undergo BMD to screen for osteopenia. Weight loss in NAFLD 

obese patients might have deleterious effects on bone health. 

Table 1. Comparison between BMI and bone mineral density in 

NAFLD patients 

Disclosures: 

The following authors have nothing to disclose: Maria Farrugia, Martina Gerada, 

John Bonello, James Mario Gauci, Richard Pullicino, Jurgen Gerada


2252 

Testosterone Is Associated with Biochemical Markers of 

Non Alcoholic Fatty Liver Disease (NAFLD) in Women 

With Polycystic Ovarian Syndrome (PCOS) 

Monika Sarkar, Chia-Ning Kao, Norah Terrault, Phyllis Tien, Ruth 

Greenblatt, Marcelle I. Cedars, Heather Huddleston; University of 


Background: Non-alcoholic fatty liver disease (NAFLD) affects 

at least half of all women with PCOS. This may relate to the 

high prevalence of insulin resistance (IR) and obesity in PCOS. 

PCOS is also characterized by hyperandrogenism although 

it is unclear whether this resting high androgen state further 

promotes worsening NAFLD in PCOS. Methods: This is a cross 

sectional study of women without heavy alcohol use and with 

confirmed PCOS (n=149) based on established Rotterdam Criteria, 

evaluated in a multidisciplinary PCOS clinic between 

2006-2014. We aimed to determine if total testosterone is 

associated with elevated ALT as a marker of NAFLD, after 

adjusting for age, body mass index (BMI) and Homeostasis 

Model Assessment of Insulin Resistance (HOMA-IR). Linear 

regression was used to evaluate the association between total 

testosterone and ALT levels, with point estimates reflecting beta 

coefficients. Results: The mean age was 28.4 (+/-6.6), median 

BMI was 28.5 (IQR 24.1-36.0), and median HOMA-IR was 

1.9, (IQR 1.0-4.7). Median ALT was 21 (IQR 14-32) and half 

of women had ALT > 1.5 times upper limits (>29 U/L). Median 

total testosterone was 54ng/dL (IQR 40-66). On unadjusted 

analysis, total testosterone was positively associated with ALT 

(beta coefficient 0.1047, 95% CI 0.0023-0.207, p=0.045) 

with stronger association after adjusting for age, BMI, and 

HOMA-IR (beta coefficient 0.1152, 95% CI 0.016-0.21, 

p=0.02). Increasing BMI (beta coefficient 0.576, 95% CI 0.18- 

0.005, p=0.005), but not HOMA-IR (beta coefficient 0.0227, 

95% CI -0.122-0.575, p=0.2) was also independently associated 

with elevated ALT. Conclusion: Elevated ALT levels, a biochemical 

marker of NAFLD, is common in women with PCOS. 

Total testosterone was associated with increasing ALT, independent 

of BMI and insulin resistance. These findings support an 

important role of androgens as a potential mediator of risk for 

NAFLD in PCOS, which may offer potential targets for NAFLD 

treatment in this high-risk population. 

Disclosures: 




The following authors have nothing to disclose: Monika Sarkar, Chia-Ning Kao, 

Phyllis Tien, Ruth Greenblatt, Marcelle I. Cedars, Heather Huddleston 

2253 

Prevalence of hepatic steatosis in healthy health care 

workers as quantified by controlled attenuation parameter 

(CAP) 

Anita Arslanow, Frank Lammert, Caroline S. Stokes; Department 

of Medicine II, Saarland University Medical Center, Homburg, 

Germany 

Background: Hepatic fat accumulation is the hallmark of non-alcoholic 

fatty liver (NAFL), for which prevalence estimates range 

widely from 5 to 50% in the population due to differences 

of screening and detection strategies. Since up to 25% of 

patients with NAFLD might develop advanced liver fibrosis 

and cirrhosis, which increase the risk of liver-associated mortality, 

better NAFL screening strategies are needed. Our aim 

was to assess the frequency of hepatic steatosis in participants 

without a history of chronic liver disease who were recruited 

from the Occupational Health Department at Saarland University 

Medical Center. Methods: For this prospective study, 81 

participants volunteered (80% women, median age 29 years, 

range 21 – 63 years). Hepatic steatosis was assessed using 

controlled attenuation parameter (CAP), which quantifies the 

degree of ultrasound attenuation based on vibration- controlled 

transient elastography (VCTE; Fibroscan). Body composition 

was determined using an eight electrode bioelectrical impedance 

analyzer (BIA). Liver function tests (LFT) and serum lipid 

concentrations were measured with standard clinical assays. 

Results: The median CAP score was 226 dB/m (range 100 

– 400 dB/m). In total, 33 participants (41%) presented with 

hepatic steatosis, when using 238 dB/m as cut-off for liver fat 

accumulation (Sasso Ultrasound Med Biol 2010). The median 

body mass index was significantly (P < 0.001) higher in participants 

with than without liver steatosis (26 vs. 22 kg/m 2 , 

respectively) and accordingly, the majority of NAFL patients 

were overweight (42%) or obese (15%). In contrast, only 15% 

of participants without hepatic steatosis were overweight, and 

none was obese (P < 0.0001). In line with these associations, 

body fat mass as determined by BIA was elevated in 48% of 

participants with hepatic steatosis, as compared to only 6% in 

those with low liver fat contents. In contrast, LFT and serum lipid 

parameters did not differ significantly between the two groups. 

Conclusions: Presence of hepatic steatosis was documented 

in almost half of this cohort of perceived healthy individuals. 

Whereas serum surrogate markers did not differentiate the individuals, 

body composition reflected the differences in liver fat 

contents among the two groups. CAP (and BIA) represent rapid 

non-invasive methods that facilitate the identification of fatty 

liver. Broad NAFL screening in the workplace might be encouraged, 

since it may allow timely detection of at risk individuals 

and early implementation of lifestyle changes. 

Disclosures: 

The following authors have nothing to disclose: Anita Arslanow, Frank Lammert, 

Caroline S. Stokes 

2254 

Assessment of parameters related to choline metabolism 

in non-alcoholic fatty liver disease: a comparison with 

healthy controls 

Hannah Da Silva 1,2 , Bianca M. Arendt 1 , Anastasia Teterina 1 , Sandra 

E. Fischer 3 , Scott Fung 4 , Johane P. Allard 4 ; 1 Gastroenterology, 

Toronto General Hospital, Toronto, ON, Canada; 2 Department of 

Nutritional Sciences, University of Toronto, Toronto, ON, Canada; 

3 Pathology, Toronto General Hospital, Toronto, ON, Canada; 

4 Medicine, Toronto General Hospital, Toronto, ON, Canada 

Background/Objectives: Choline deficient diets promote steatosis 

similar to non-alcoholic fatty liver disease (NAFLD). Thus, 

one of the proposed mechanisms for NAFLD pathogenesis is 

reduced choline bioavailability resulting from the metabolism 

of dietary choline to trimethylamine (TMA) by gut micriobiota, 

causing subsequent reduction of phosphatidylcholine in the 

liver and decrease in VLDL export of triglycerides. The validity 

of this proposed mechanism was tested in animal models, 

but data on choline metabolism in general NAFLD population 

and its relation to intestinal microbiota is lacking. Our objective 

was to assess parameters related to choline metabolism 

and determine whether these parameters are associated with 

NAFLD. Methods: We studied serum choline and fecal choline 

and TMA in 39 biopsy-confirmed patients (15 simple steatosis 

(SS), 24 non-alcoholic steatohepatitis (NASH)), and 28 living 

liver donors as healthy controls (HC) by nuclear magnetic resonance 

spectroscopy. Clinical, biochemical, and anthropometric 

data were also collected. Dietary intake was assessed using 

a 7-day food record in a subset of participants (10 SS, 20


NASH, 25 HC). The data are presented as median (Q1, Q3) 

for non-normal and mean (SD) for normal continuous variables. 

Groups were compared using Wilcoxon rank-sum tests and 

t-tests. Spearman correlation coefficients were used to examine 

association between variables. Results: NAFLD patients were 

older than HC and had significantly higher BMI, liver enzymes 

(AST, ALT), insulin resistance (HOMA-IR), hemoglobin A1c, 

and blood triglycerides (p


(5%) pt had uncontrolled diarrhea requiring hospitalization. 

Other minor reported AE’s were fatigue (20%), anemia (15%), 

rash/itching (10%) and nausea (5%). Conclusion In summary, 

HCV in ESRD is considered a difficult-to-treat group and in 

whom SVR is associated with overall clinical improvement and 

outcomes after kidney transplantation. Half-dose SOF based 

regimens provides an attractive option. The regimen appears 

to be a safe, well-tolerated and efficacious resulting in higher 

rates of sustained virologic response. Additional SVR 12 will 

be available for presentation. 

Demographics 

Grade 2 event (body aches in a patient receiving 100 mg EDP- 

239). There were no serious adverse events. The mean terminal 

half-life for EDP-239 was approximately 24 hours; inter-subject 

variability as measured by CV% was


treatment of chronic Hepatitis C Virus infection. TT-034 uses the 

process of DNA directed RNAi interference (ddRNAi) which 

triggers the cell’s own transcriptional machinery to continuously 

produce steady state levels three independent short hairpin 

RNA (shRNA) to target 3 independent regions of the HCV 

genome. Designed to be administered as a single treatment 

delivered intravenously, TT-034 uses an Adeno-Associated 

Virus type 8 (AAV8) capsid to deliver a recombinant genome 

preferentially into hepatocytes. Biodistribution analyses presented 

here demonstrate that 90% of the vector distributes 

into liver tissues, with close to 100% transduction of the liver 

hepatocytes, as assessed by in situ hybridization. Furthermore, 

the durability to expression following a single injection, as 

assessed by qPCR, demonstrated that shRNA expression persisted 

for the duration of the 180 day experiment. Because 

previous reports have suggested that high expression of shRNA 

may cause global dysregulation of endogenous miRNA processes 

within cells, the impact of long term expression of TT-034 

on endogenous miRNA levels was interrogated. Analyses were 

performed using RNA isolated from liver biopsies at Day 15, 

and from liver and heart tissues collected 60 or 180 days 

post TT-034 administration. This miRNA profiling demonstrated 

reliable detection of 260 microRNAs in the primate heart samples, 

as compared to 266 in liver and 269 in the liver biopsy 

samples. The analyses of heart tissues demonstrated that there 

was no statistical difference across the groups treated with 

TT-034 (ANOVA Benjamini Hochberg (BH) pvalue < 0.05). 

Although the liver biopsy samples showed significant effects in 

27 microRNA, analyses of the day 60 and day 180 liver samples 

showed no statistical differences from the control group. 

Collectively, these data suggest that TT-034 is not likely to have 

any adverse effects in miRNA processes in primate cells. 

Disclosures: 

David Suhy - Advisory Committees or Review Panels: Regen BioPharma; Management 

Position: Benitec Biopharma 

Tin Mao - Employment: Benitec Biopharma 

Shih-Chu Kao - Employment: Benitec Biopharma 

Per Lindell - Consulting: Benitec Ltd, Tacere Inc 

2259 

Pharmacokinetics, Safety, and Tolerability of EDP-239, 

a Novel Hepatitis C NS5A inhibitor, in Healthy Volunteers 

Lijuan Jiang 2 , Xuemin Jiang 1 , Kiran Dole 1 , Kimberley Caliri 2 , Kenneth 

Tack 2 , Richard Colvin 1 , Yat Sun Or 2 ; 1 NIBR, Cambridge, MA; 

2 Enanta Pharmaceuticals, Watertown, MA 

Background: EDP-239, a novel NS5A inhibitor, demonstrates 

potent anti-HCV activity in vitro, with mean EC 50 

values of 31 

and 10 pM against genotypes 1a and 1b, respectively. Methods: 

Pharmacokinetics, safety, and tolerability of EDP239 were 

assessed in a randomized, double-blind, placebo-controlled 

study. 94 healthy volunteers received EDP239/placebo either 

as single oral doses from 3 mg to 300 mg or 7 days of QD 

dosing of 10 mg to 200 mg. Results: EDP239 plasma exposure 

(C max 

and AUC) increased with dose in a slightly more 

than dose-proportional manner following single doses or 7-day 

dosing. The half-life was approximately 24 hours. Steady-state 

was achieved by Day 5 with a 2-fold increase in exposure. On 

Day 7, inter-subject variation for AUC 0-24 

was less than 22% 

from 10 mg through 200 mg; mean AUC 0-24 

was 73,500 

ng*hr/mL at the 200 mg dose. EDP239 was well tolerated. 

Rates of adverse events (AEs) were similar in EDP239 and 

placebo-treated subjects. There were no serious adverse events 

and no discontinuations due to AEs. No clinically significant 

laboratory abnormalities were observed. Conclusions: EDP239 

was well absorbed, achieving therapeutic levels at achievable 

doses. No safety or tolerability issues were observed. Further 

development of EDP239 as part of a combination regimen for 

HCV infection is warranted. 

EDP239 plasma PK parameters following QD oral dosing x 7d in 

healthy volunteers 

Disclosures: 

Lijuan Jiang - Employment: Enanta Pharmaceuticals, Inc 

Xuemin Jiang - Employment: Novartis 

Kenneth Tack - Consulting: Enanta Pharmaceuticals, Inc 

Richard Colvin - Employment: Novartis Institute for Biomedical Research 

Yat Sun Or - Employment: Enanta Pharmaceuticals, Inc, Enanta Pharmaceuticals, 

Inc 

The following authors have nothing to disclose: Kiran Dole, Kimberley Caliri 

2260 

Pharmacokinetics, Safety and Tolerability of EDP-239, a 

Novel Hepatitis C (HCV) NS5A inhibitor, in Patients with 

HCV Infection 

Lijuan Jiang 1 , Xuemin Jiang 2 , Kiran Dole 2 , Eric Lawitz 3 , Fred Poordad 

3 , Stefan Zeuzem 4 , Kimberley Caliri 1 , Kenneth Tack 1 , Richard 

Colvin 2 , Yat Sun Or 1 ; 1 Enanta Pharmaceuticals, Watertown, 

MA; 2 Novartis Institutes for Biomedial Research, Cambridge, MA; 

3 Texas Liver Institute, San Antonio, TX; 4 Johan Wolfgang Goethe 

Universitat, Frankfurt, Germany 

Background: EDP-239, a novel NS5A inhibitor, demonstrates 

potent anti-HCV activity in vitro. Studies in normal volunteers 

have shown good absorption, safety, and tolerability at doses 

up to 200 mg daily x 7 days. Methods: Pharmacokinetics, 

safety, and tolerability of EDP-239 were assessed in a randomized, 

double-blind, placebo-controlled study. 28 patients with 

chronic HCV (GT 1a/b) infection received EDP-239 or placebo 

as single oral doses from 10 mg to 200 mg. Results: EDP- 

239 was rapidly absorbed following a single oral dose with 

T max 

of 2 – 3 hrs. Plasma exposure (C max 

and AUC) increased 

with dose in a slightly more than dose-proportional manner 

following single doses from 10 mg to 200 mg. The half-life 

was approximately 24 hours. EDP-239 was well tolerated. 

Rates of adverse events (AEs) were similar in EDP239- and 

placebo-treated patients. Almost all AEs were mild in intensity. 

There were no serious adverse events. No clinically significant 

laboratory abnormalities were observed. Conclusions: EDP- 

239 was well absorbed, achieving supratherapeutic levels in 

HCV-infected patients. Mean plasma concentration 24 hrs after 

a single oral dose of 100 mg was > 10,000 X the in vitro 

replicon EC 50 

values of genotype 1 virus. EDP-239 was well 

tolerated and no safety issues were observed. Further development 

of EDP-239 as part of a combination regimen for HCV 

infection is warranted.


EDP-239 plasma PK parameters following oral dosing in HCV 

patients 

Disclosures: 

Lijuan Jiang - Employment: Enanta Pharmaceuticals, Inc 
















Kenneth Tack - Consulting: Enanta Pharmaceuticals, Inc 


Yat Sun Or - Employment: Enanta Pharmaceuticals, Inc, Enanta Pharmaceuticals, 

Inc 

The following authors have nothing to disclose: Kiran Dole, Kimberley Caliri 

2261 

Short-term safety and pharmacokinetics of co-administration 

of EDP239, an HCV NS5A inhibitor, and alisporivir 

(ALV), a cyclophilin inhibitor, in healthy subjects 

Xuemin Jiang 1,2 , Richard Colvin 1,2 , June Ke 1,2 , Sachin Desai 1,2 , 

Surendra Machineni 1,2 , Jie Zhang 1,2 , Wei Zhou 1,2 , Sun Haiying 

1,2 , Steven J. Kovacs 1,2 ; 1 Novartis, East Hanover, NJ; 2 Institutes 

for Biomedical Research, East Hanover, NJ 

Introduction: Cyclophilin A plays essential roles in HCV replication 

and assembly, including modulating the function of NS5A. 

Combined cyclophilin and NS5A inhibition is a clinically 

untested therapeutic approach despite in vitro studies suggesting 

additive-to-synergistic effects of the two mechanisms, acting 

at functionally dependent sites of the HCV replication complex. 

An EDP239 C 24h 

of 317 ng/mL reduced viral load by >3 

log10 after a single 100 mg dose in GT1-infected patients. ALV 

is a cyclophilin inhibitor that has pan-genotypic anti-HCV activity 

with demonstrated clinical efficacy. It is a time-dependent 

CYP3A4 inhibitor and inhibitor of multiple uptake and efflux 

transporters, including P-gp. EDP239 is a substrate of CYP3A4 

and P-gp in vitro. Methods: Open-label; fixed sequence design 

(n=26). Subjects received multiple doses of EDP239 30 mg BID 

for 21 days with co-administration of ALV 400 mg BID days 8 

through 21. Both drugs administered with food. Blood samples 

were collected up to 12 hours after dose administration on 

days 7, 15 and 21 to measure plasma concentrations by LC/ 

MS/MS. Non-compartmental PK analysis using WinNonlin; 

ANOVA for statistical comparisons; routine safety assessments 

with safety data descriptively summarized. Results: 22 subjects 

completed the study. Study treatments were well tolerated 

with no safety-related discontinuations. Adverse effects were 

consistent with the known effects of either EDP239 (gastrointestinal 

complaints) or ALV (gastrointestinal complaints, headache, 

laboratory abnormalities, increases in blood pressure). 

Median EDP239 T max 

was 3, 4.5, and 4 h on days 7, 14, 

and 21, respectively. The EDP239 C 12h 

was 2180 ng/mL 

when administered with ALV. EDP 100 mg was comparable 

to C12h of EDP 30 mg when combined with a alisporivir. 

EDP239 had no apparent effect on ALV exposure (compared 

with historical data). Conclusion Short term co-administration 

of EDP239 and ALV was safe and well tolerated. The data 

suggest that therapeutic combination of ALV and EDP239 is 

feasible. The observed EDP239 exposures can be expected 

to have clinical activity against GT1 and GT3 HCV. The time 

course of exposure to EDP239 was relatively flat with minimal 

difference between peak and trough. Exploiting the interaction 

may allow EDP239 to treat patients with G3 HCV infection 

without increasing overall exposure beyond exposures previously 

safely investigated in humans. 

Disclosures: 



Surendra Machineni - Employment: Novartis Healthcare Private Limited 

Steven J. Kovacs - Employment: Novartis 

The following authors have nothing to disclose: June Ke, Sachin Desai, Jie Zhang, 

Wei Zhou, Sun Haiying 

2262 

SB 9200 Shows Antiviral Activity Against HCV-RNA 

by Targeting Host Cytosolic Sensor Proteins RIG-I and 

NOD-2 to activate IFN signaling pathways 

Kumar Visvanathan 2,1 , Rosemary M. Millen 2,1 , Stuart K. Roberts 4,5 , 

Peter W. Angus 6,2 , Wendy Cheng 8 , Nada Farhat 9 , My My Trinh 9 , 

Radhakrishnan P. Iyer 10 , Murray L. Barclay 7 , Alex J. Thompson 3,2 ; 

1 Innate Immunity Laboratory, St Vincent’s Hospital Melbourne, Melbourne, 

VIC, Australia; 2 Medicine, Univeristy of Melbourne, Melbourne, 

VIC, Australia; 3 Gastroenterology, St. Vincents Hospital, 

Melbourne, Fitzroy, VIC, Australia; 4 Gastroenterology, Alfred Hospital, 

Melbourne, VIC, Australia; 5 Medicine, Monash University, 

Melbourne, VIC, Australia; 6 Liver Transplant Unit, Austin Hospital, 

Melbourne, VIC, Australia; 7 Gastroenterology & Clinical Pharmacology, 

Christchurch Hospital and University of Otago, Christchurch, 

New Zealand; 8 Gastroenterology, Royal Perth Hospital, 

Perth, WA, Australia; 9 Pharsight Consulting Services, Princeton, 

NJ; 10 Spring Bank Pharmaceuticals, Milford, MA 

Introduction. SB9200 activates RIG-I and NOD-2 pathways in 

virally infected cells and has been shown to maximally reduce 

HCV RNA by 1.9 log10 in HCV patients (Thompson EASL 

2015). We aimed to evaluate the relationship between the 

pharmacokinetics of SB 9200, and the induction of innate 

immunity and antiviral response. Methods. HCV patients, GT 1 

and 3, received 200-900 mg of SB9200 PO daily for 7 days. 

Early antiviral kinetics were measured from baseline through 

7 days after the first dose of SB 9200. Patients were divided 

into null responders at day 7 (NR, < 0.5 log10 reduction in 

HCV RNA, n=11) or responders (R, >0.9 log10, n=6). Clinical 

characteristics at baseline for NR vs. R were similar for 

median HCV RNA, BMI, ALT, AST, platelets and Fibroscan. 

50% of R were IL28b CC compared to 18% of NR. Plasma 

interferon alpha (IFNa) was measured on day 1, 7 and 14 

by ELISA. Interferon stimulating genes ISG15 and OAS1 were 

measured on Day 1, 2, 3, 7 and 14 in peripheral blood via 

RT-PCR. GAPDH was used as an internal control. Data are represented 

as ratios from baseline for IFNa, ISG15, and OAS1. 

For pharmacokinetic analysis, blood samples were collected on


Day 7 from pre-dose through 48 hrs post-dose. Results. Plasma 

IFNa was significantly increased from baseline in R vs.NR 

(Fig 1; P


HCV replication and the impact on the response to treatment of 

HCV Materials and Methods: The replicon cells Huh7/Ava.5 

(genotype 1b), Huh7/J6/JFH (genotype 2a) and Huh7.5/ 

Con1 (genotype 1b) were obtained and the mirVana let-7g 

mimic/inhibitor, miR-122 inhibitor and miRNA mimic/inhibitor 

negative control were purchased. The 1.0-kb and 0.5- 

kb fragments of the let-7g promoter were amplified by PCR. 

Site-directed mutagenesis of the AP-1 binding-site presented in 

the let-7g promoter region was carried out. WST-1 assay and 

Renilla Luciferase Assay were used. Expression levels of let-7g 

in each sample were normalized to the corresponding level of 

snU6B. Expression levels of let-7g were determined by using 

the Quantitative real-time PCR. Anti-phospho-ERK, p38, JNK, 

and total-ERK, p38, JNK antibodies, Anti-GAPDH and α-tubulin 

antibodies were used for Immunoblot analysis. Results: 

Our results demonstrated that overexpression of let-7g reduces 

HCV expression in the cell line. The HCV loads were more 

decreased by let-7g mimic than miR-122 inhibitor transfected 

cell. High levels of lin28 correlate with low levels of let-7g 

in HCV-infected cells and the knockdown of lin28 via siRNA 

reduces HCV replication. The treatment with a let-7g mimic 

alone was shown to induce IFN-induced genes inclusion MxA 

and OAS1. Interferon (IFN)/RBV induces let-7g expression, 

and let-7g and IFN/ribavirin also elicited an addictive inhibitory 

effect on HCV expression. The anti-viral effects of let-7g 

mediated IFN/RBV signaling and the regulation of let-7g by 

IFN/RBV occurs through p38/AP1 signaling. Conclusions: We 

have indicated an important role of let-7g on the replication of 

HCV and on the response of HCV to anti-HCV treatment. The 

IFN/ribavirin induces let-7g expression through p38/AP-1 signaling 

and the let-7g and IFN/RBV have additively inhibitory 

effect on HCV replication. The let-7g may serve as a potential 

target for HCV therapy. 

Disclosures: 



Ming-Lung Yu - Advisory Committees or Review Panels: ABBOTT, MSD, ABBVIE, 

GILEAD, J&J, ROCHE, BMS; Grant/Research Support: ABBOTT, ROCHE, MSD, 

ABBVIE, GILEAD; Speaking and Teaching: ABBOTT, ROCHE, MSD, GILEAD, 

BMS, GSK 

The following authors have nothing to disclose: Chia-Yen Dai, Wen-Wen Chou, 

Yi-Shan Tsai, Shu-Chi Wang, Chung-Feng Huang, Ming-Lun Yeh, Jee-Fu Huang 

2265 

Drug-Drug Interaction Studies between HCV Antivirals 

Sofosbuvir and GS-5816 and HIV Antiretrovirals 

Erik Mogalian 1 , Luisa M. Stamm 2 , Anu Osinusi 2 , Gong Shen 4 , 

Karim Sajwani 3 , John McNally 2 , Anita Mathias 1 ; 1 Clinical Pharmacology, 

Gilead Sciences, Inc., Foster City, CA; 2 Clinical Research, 

Gilead Sciences, Inc., Foster City, CA; 3 Clinical Operations, Gilead 

Sciences, Inc., Foster City, CA; 4 Biostatistics, Gilead Sciences, 

Inc., Foster City, CA 

Introduction A once-daily fixed-dose combination tablet composed 

of sofosbuvir (SOF; nucleotide analog NS5B inhibitor) 

and GS-5816 (pangenotypic NS5A inhibitor) is in clinical 

development for the treatment of chronic HCV infection. Phase 

1 studies were conducted in healthy volunteers to evaluate 

potential drug-drug interactions (DDIs) between SOF/GS-5816 

and HIV antiretroviral (ARV) regimens without a pharmacokinetic 

“booster” (ritonavir or cobicistat) to support their use 

together in HIV/HCV co-infected patients. Methods These were 

multiple-dose, randomized, cross-over DDI studies. Subjects 

received SOF/GS-5816 and ARVs (EFV/FTC/TDF [Atripla ® ; 

ATR], FTC/RPV/TDF [Complera ® ; CPA], DTG [Tivicay ® ], and 

RAL [Isentress®] + FTC/TDF [Truvada®]) alone and in combination. 

Steady-state plasma concentrations of SOF, its predominant 

circulating nucleoside metabolite GS-331007, GS-5816, 

and ARVs were analyzed on the last day of dosing for each 

treatment and PK parameters were calculated. Geometric leastsquares 

means ratios and 90% confidence intervals (combination 

vs. alone) for SOF, GS-331007, GS-5816, and ARV 

AUC tau 

, C max 

and C tau 

were estimated and compared against 

pre-specified lack of PK alteration boundaries of 70-143% for 

all analytes except RAL (50-200%). Safety assessments were 

conducted throughout the study. Results Twenty-nine of 30 subjects 

in Group 1 and all subjects in Groups 2-4 (N=78 total) 

completed the study. The majority of adverse events (AEs) were 

Grade 1 and there were no serious AEs. The most frequent AEs 

were headache (12%) and constipation (10%). One subject in 

Group 1 discontinued due to an AE (Grade 1 urticaria). SOF/ 

GS-5816 PK was unaffected by RPV, DTG, and RAL regimens. 

A decrease (~53%) in GS-5816 exposure with no impact on 

overall SOF or GS-331007 exposure was observed following 

administration with ATR. No clinically significant changes 

in the PK of DTG, EFV, FTC, RAL, and RPV were observed 

when administered with SOF/GS-5816. Increased TFV exposure 

(ATR: ~1.8-2.2-fold; CPA: ~1.4-1.8-fold; FTC/TDF+RAL: 

~1.4-1.7-fold) was observed with SOF/GS-5816; overall, 

absolute TFV AUC in the test (combination) treatments were 

similar to those achieved when FTC/TDF is administered with 

ritonavir-boosted PIs, which do not warrant dose adjustment. 

Conclusion Study treatments were generally well tolerated. 

Results from this study demonstrate that SOF/GS-5816 may be 

administered safely with CPA, RAL and DTG with a backbone 

of FTC/TDF. Additional data from SOF/GS-5816 Phase 3 PK/ 

PD will guide the recommendation for use with ATR. 

Disclosures: 

Erik Mogalian - Employment: Gilead Sciences, Inc; Stock Shareholder: Gilead 

Sciences, Inc 


Anu Osinusi - Employment: Gilead Sciences 

Karim Sajwani - Employment: Gilead Sciences, Inc. 

John McNally - Employment: Gilead Sciences, Inc; Stock Shareholder: Gilead 

Sciences, Inc 


The following authors have nothing to disclose: Gong Shen 

2266 

Discovery of AT-337 and AT-339, Two Highly Potent 

and Selective Nucleotide Prodrug Inhibitors of HCV 

Polymerase 

Steven S. Good 1 , Adel Moussa 1 , Jean-Christophe Meillon 2 , Jean- 

Pierre Sommadossi 1 ; 1 Atea Pharmaceuticals Inc., Boston, MA; 

2 Oxeltis, Montpellier, France 

Background: Nucleotide analogs are preferred candidates for 

treatment of HCV infection since they are potent, pan-genotypic 

inhibitors of NS5B polymerase with a high barrier to 

drug resistance. Here we report novel nucleotide prodrugs with 

both base and sugar modifications that are selective for and 

highly active against in vitro HCV replication. Methods: Novel 

nucleotide prodrugs were synthesized and tested in Huh-7 cells 

bearing an HCV genotype 1b replicon and evaluated for activity 

against other viruses. Selectivity was assessed in 14-day 

exposure with human BFU-E and CFU-GM cells and in 3-day 

exposure with human iPS cell-derived cardiomyocytes. Selected 

nucleoside-5’-triphosphate analogs were evaluated against the 

NS5B polymerase and human DNA polymerases α, β and 

γ. Results: The two most potent nucleotide prodrugs, AT-337 

and AT-339, inhibited HCV replication, with EC 50 

values as 

low as 0.005 mM. Intra-assay comparisons demonstrated that 

AT-337 and AT-339 were at least 10-fold more potent than


sofosbuvir, as shown in the inhibition curves below. A unique 

feature of this series is the formation of at least two active 

metabolites from each candidate that compete against different 

naturally occurring ribonucleoside triphosphates, with IC 50 

values 

against the NS5B polymerase between 1.7 and 0.11 mM. 

AT-337 and AT-339 demonstrated excellent selectivity, with 

CC 50 

values greater than 100 mM in Huh-7 cells, human bone 

marrow stem cells and human cardiomyocytes. Furthermore, 

no inhibition of human DNA polymerase α, β or γ, no activity 

against other RNA or DNA viruses, and no toxicity in all host 

cell lines was observed at concentrations up to 100 mM. Conclusions: 

New nucleotide prodrugs potently inhibited the HCV 

NS5B polymerase by producing multiple active analogs of different 

ribonucleoside triphosphates. IND enabling studies are 

underway, targeting clinical evaluation in early 2016. 

Disclosures: 

Steven S. Good - Employment: Atea Pharmaceuticals Inc.; Stock Shareholder: 

Atea Pharmaceuticals Inc. 

Jean-Christophe Meillon - Employment: Oxeltis; Independent Contractor: Atea 

Pharmaceuticals, Inc.; Stock Shareholder: Oxeltis 

The following authors have nothing to disclose: Adel Moussa, Jean-Pierre Sommadossi 

2267 

The impact of α-fetoprotein level during interferon-free 

treatment of hepatitis C virus 

Masataka Seike, Koichi Honda, Junya Oribe, Mizuki Endo, Mie 

Yoshihara, Masao Iwao, Masanori Tokoro, Kazunari Murakami; 

gastroenterology, Oita University, Yufu-city, Japan 

AIM: The prevalence of older patients with hepatitis C virus 

(HCV) has been increasing in Japan. Elderly patients are at 

higher risk for hepatocellular carcinoma (HCC) even after HCV 

eradication. The risk of developing HCC is age-dependent, and 

patients aged ≥ 75 years occasionally present with HCC. α-Fetoprotein 

(AFP) levels after interferon (IFN) therapy for HCV are 

significantly associated with hepatocarcinogenesis. However, 

the significance of AFP level after IFN-free therapy in older 

patients remains unclear. Thus, we analyzed AFP levels during 

IFN-free treatment in patients with HCV. Patients and methods: 

This study included 63 patients (31 males and 32 females) 

with HCV genotype 1b who received 60-mg daclatasvir once 

daily plus 100-mg asunaprevir twice daily. The mean age of 

these patients was 71.3 years (range, 49–82 years). AFP level 

was assessed at baseline and every month. These cases were 

followed up by abdominal ultrasonography (US), contrast-enhanced 

computed tomography (CT), or ethoxybenzyl contrast 

magnetic resonance imaging (MRI) every 3–6 months. Results: 

The mean serum AFP level in all patients before treatment was 

26.7 ng/ml. AFP level during treatment tended to decrease 

immediately and had decreased to 11.7 ng/ml at the end of 

treatment. Serum AFP levels in 36 patients decreased to < 5 

ng/ml at the end of treatment (group A). The group A patients 

had no HCC on abdominal US, CT, or MRI. However, serum 

AFP levels in 27 patients decreased to > 5 ng/ml at the end of 

treatment or increased during therapy (group B). Twelve group 

B patients (44%) were diagnosed with HCC after treatment. 

Conclusion: The AFP trend during treatment was useful for predicting 

future HCC risk after all-oral therapy with daclatasvir 

plus asunaprevir for HCV. Patients with HCV and an AFP level 

> 5 ng/ml at the end of treatment should be examined for HCC 

by US, CT, and/or MRI as soon as possible. 

Disclosures: 

The following authors have nothing to disclose: Masataka Seike, Koichi Honda, 

Junya Oribe, Mizuki Endo, Mie Yoshihara, Masao Iwao, Masanori Tokoro, 

Kazunari Murakami

1312A AUTHOR INDEX HEPATOLOGY, October, 2015 

Author Index 

All numbers refer to the corresponding abstract. 

A. Bhanji, Rahima 308 

Aa, Jiye 919, 934 

Aagaard, Niels Kristian 768, 2071 

Aarslev, Kristian 301 

Abate, Maria Lorena 1740, 2209 

Abbas, Zaigham 1872 

Abbasy, Mohammed 733 

Abd, Mortadha 943 

Abdallah, Ayat 1890 

Abdel-Razek, Wael 733 

Abdelatif, Dinan 892, 2173, 2192, 

2212 

Abdelmalek, Manal F. 105, 161, 162, 

239, 737, 739, 889, 1428, 1435, 

2145, 2153, 2154, 2169, 2220, 2239 

Abdelsameea, Eman 1890 

Abe, Hiromi 663, 986, 1150, 1627, 

1645, 1649, 1670, 1679, 1681, 

1858, 1895, 1944, 2120 

Abe, Hiroshi 1097 

Abe, Masafumi 551 

Abe, Masanori 338, 788, 1293, 2146 

Abe, Mitsuhiko 688 

Abe, Yuta 336 

Abecassis, Michael M. 71 

Aberg, Judith 380, 457 

Abergel, Armand 1036 

Abergel, Armando 95, 219, 777, 1009, 

1505 

Abiru, Seigo 408, 783, 1600 

Aboelsoud, Mohammed M. 169, 1625 

Abouelhoda, Mohamed 1821 

Abouljoud, Marwan S. 556, 1274 

Aboulnasr, Fatma 501, 506, 1096 

Abraham-Enachescu, Ioana 1940 

Abraldes, Juan 742, 745 

Abramowsky, Carlos R. 12 

Abrams, Marc 2116 

Abrams, Michael 2007 

Abreu, Rodrigo M. 1544 

Abreu de Carvalho, Luis Filipe 487 

Abreu-Gonzalez, Pedro 1342 

Abrignani, Sergio 2011 

Abt, Peter L. 837, 853, 864, 868, 871 

Abu Shanab, Ahmed 1275 

Abu-Raddad, Laith 1829 

Abunimeh, Manal 722, 1039 

Abzug, Mark J. 1737 

Acalovschi, Monica 463 

Achahboun, Mohamed 480 

Achiwa, Koichi 967 

Acibucu, Fettah 2219 

Ackerman, Peter 716, 726, 728 

Acosta Umanzor, Carlos 691 

Adam, Rene 449, 487, 1237 

Adams, David H. 1296, 1298, 1710 

Adams, Leon A. 848, 1264, 2099, 

2185 

Adams, Paul 2117 

Adams, William 1172 

Adamson, Gord 424 

Addissie, Benyam D. 397 

adebajo, corlan 539 

Adelmeijer, Jelle 740, 2249 

Adem, Fatima 1118 

Adeyi, Oyedele 11 

Adhoute, Xavier 394, 2082 

Adinolfi, Luigi 1837 

Adler, Joel T. 1732 

Adorini, Luciano 142, 882, 893, 902, 

907 

Aerts, Raymond 171 

Afdhal, Nezam H. 96, 249, 737, 739, 

746, 1130, 1428, 1435, 1442, 2227 

Afdhal, Sophie K. 1177 

Afendy, Mariam 534 

Afonso, Marta B. 176, 832, 897, 940, 

960 

Afredj, Nawel 1621 

Africa, Jonathan A. 159 

Afsharzadeh, Danial 1416 

Agarwal, Amol 536 

Agarwal, Banwari 1769, 1780, 1781 

Agarwal, Kosh 249, 646, 1084, 1098, 

1113, 1117, 1132, 1173, 1588, 

1704, 1744, 2053, 2054 

Agarwal, Parul D. 434 

Agarwal, Rajiv 267 

Agarwal, Ritu 1095, 1185 

Agarwal, Shaleen 853 

Aggarwal, Rakesh 1475, 1817 

Aggarwal, Richa 1945 

Aghemo, Alessio 1743, 1747 

Agiasotelli, Danai 1489, 2096 

Agopian, Vatche 369 

Agostinelli, Laura 21 

Agrawal, Saurabh 1903 

Agrawal, Swastik 214 

Aguiar, Miguel Osman D. 1484, 1494, 

1495 

Aguilar, Humberto I. 41, 1065 

Aguilar, Maria 237, 1252, 2159 

Aguilar Schall, Raul E. 624, 632, 737, 

739, 746, 777, 1428, 1435, 1442, 

2149, 2239 

Aguilar-Urbano, Víctor 2171 

Aguilera, Victoria 1230, 1251 

Ahammed, Sk Mahiuddin 750 

Ahlén, Gustaf 992 

Ahlquist, David 169 

Ahluwalia, Vishwadeep 52, 1472, 

1491 

Ahmad, Dina 748, 769 

Ahmad, Irfan 745 

Ahmad, Jawad 1185, 1922, 1931 

Ahmad, Maliha 295, 1181 

Ahmed, Aijaz 199, 201, 237, 534, 

541, 877, 1079, 1182, 1189, 1252, 

1534, 1538, 1745, 1750, 2155, 2159 

Ahmed, Emilia 2072, 2076 

Ahmed, Emily 1242 

Ahmed, Kazi 316 

Ahmed, Ola 1821 

Ahmed, Osman 542 

Ahmed, Shahzad 1279 

Ahmed, Zohair 774 

Ahmed Mohammed, Hager 381, 415 

Ahn, Hongkeun 242, 479, 2016 

Ahn, Jem Ma 433, 1504, 1591 

Ahn, Joseph 1101 

Ahn, Joseph C. 7 

Ahn, Sang Bong 924, 941, 951, 2218 

Ahn, Sang Hoon 241, 346, 504, 1542, 

1558, 1581, 2014, 2025 

Ahn, Daegeon 423 

Ahrens, William A. 653, 1023 

Ahuja, Chirag K. 214 

Ahya, Vivek N. 536 

Aibe, Yuki 760, 1447, 1464, 1492 

Aichelburg, Maximilian C. 561, 1204 

Aihara, Arihiro 1962 

Aihara, Eitaro 676 

Aikata, Hiroshi 472, 986, 1056, 1150, 

1627, 1649, 1670, 1681, 1858, 

1893, 1895, 1944, 2120, 2158 

Ain, Dani 1076 

Ait Younes, Sonia 1621 

Ait-Oufella, Hafid 2148 

Aït-Slimane, Tounsia 196 

Aithal, Guruprasad P. 225, 908, 1524, 

1930, 1933 

Aitken, Celia 1803 

Aizawa, Nobuhiro 466, 770, 781, 

1533, 1669, 1975 

Ajmera, Veeral H. 2161 

Akamatsu, Sakura 986, 1649, 1681, 

1858, 1944 

Akarca, Ulus S. 246, 558, 1247, 1739, 

1822 

Akat, Kemal M. 1364 

Akazawa, Yoshihiro 1956 

Akazawa, Yuko 952, 1671 

Akbani, Rehan 130 

Akbar, Sheikh Mohammad Fazle 2157 

Akiba, Jun 465, 1990 

Akihiro, Isogawa 2240 

Akinc, Akin 36 

Akita, Maaya 906, 2156 

Akiyama, Takumi 296 

Akpadock, Evelyn 197 

Akremi, Raoudha 206 

Akriviadis, Evangelos 1126 

Akuta, Norio 1062, 1191, 2020, 2241 

Akyuz, Filiz 478 

Akyuz, Umit 478 

Al Ikhwan, Mahdi 1431 

Al Kaabi, Saad R. 1843 

Al Mamari, Said 622 

Al Mamun, Ayub 2157 

Al Masaoudi, Malika 1924 

Al Zoairy, Ramona 1092, 1885 

Al-Akkad, Walid 337, 1388 

Al-ashgar, Hamad I. 1607 

Al-Ejji, Khalid 1188 

Al-Judaibi, Bandar 1246 

Al-khafaji, Ali 275 

Al-Khatib, Safa H. 113 

Al-mana, Hadeel 1607 

Al-Nujaidi, Danya Y. 413 

Al-Osaimi, Abdullah M. 765, 766, 

1122 

Al-Rubaish, Fatima A. 413 

Alahdab, Fares 282 

Alain, Sophie 1162 

Alam, Altaf 1872 

Alam, Seema 1697, 1725 

Alao, Hawwa 218, 1498

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) AUTHOR INDEX 1313A 

Alaparthi, Lakshmi 892, 1428, 2173, 

2192 

Alarcon-Galvan, Gabriela 1281 

Alarcón-Vila, Cristina 899 

Alatrakchi, Nadia 25, 1015 

Alavi, Maryam 543, 1868 

Albeladi, Khalid 1607 

Albert, Dustin 285 

Alberti, Alfredo 775, 1089, 1802, 

1844 

Alberts, Steven 127, 1966 

Albillos, Agustin 745, 2171 

Albrecht, Jeffrey 31 

Albuerne, Marisol 2100 

Albuquerque, Miguel 480 

Alcântara, Camila M. 2072, 2076 

Alcaraz-Quiles, Jose 2087 

Aldairy, Wassim 1920 

Aldea, A. 596 

Alegre, Fernando 1913 

Alessandra, Mazzola 1239 

Alessio, Loredana 1543 

Àlex, Amorós 2069 

Alexander, Graeme J. 258 

Alexandrino, Paula 150 

Alexopoulou, Alexandra 1489, 2019, 

2096 

Alfadda, Abdulrahman A. 1843 

Alghanem, Mansour G. 1246 

Alharbi, Asia 1118 

Alhassani, Abdulla 1118 

Alhasson, Firas 962, 2133, 2137 

Ali, Irfan 2173, 2192 

Alibrandi, Angela 288, 1898 

Alison, Malcolm 101 

Aljudaibi, Bandar 204 

Aljumah, Abdulrahman A. 1607 

Alkaabi, Saad R. 1147, 1188 

Alkhatib, Zahraa 344 

Allaire, Manon 2082 

Allard, Johane P. 2184, 2238, 2254 

Allard, Nicole 2060 

Allawi, Hussain 892 

Allawy, Allawy 330, 334, 518, 651, 

667, 668 

Allen, Alina M. 6, 2155 

Aller, Rocío 2171 

Allotey, Loretta K. 1965, 1966 

Almagro, Jorge 1295, 1520 

Almarzooqi, Saeed 1099 

Almazrouei, Sultan 1118 

Almeida, Marcio D. 469 

Almeida-Dalmet, Swati S. 1527 

Almer, Sven 76 

Almishri, Wagdi 1290 

Aloman, Costica 1266, 1294, 1370, 

2124 

Alonso, Cristina 953, 2163, 2244 

Alonso, Estella M. 133, 1714 

Alonso-Guallart, Paula 2129 

Aloysius, M. 1037, 1074, 1075 

Alpini, Gianfranco 190, 602, 621, 817, 

834, 835, 929, 1317, 1349, 1365, 

1978 

Alqahtani, Saleh 1057 

Alric, Laurent 95, 1158, 1803, 2082 

Alsaad, Khaled 1607 

Alsahhar, Jamil S. 1199 

Alsebaey, Ayman 1862, 1879 

Alsina, Angel 526, 1784 

Alswat, Khalid A. 1607 

Alt, Yvonne 1949 

Altamirano, José T. 110, 1307, 1327, 

2148 

Althouse, Sandra K. 436 

Altice, Frederick 40 

Altinkaya, Engin 2219 

Altintas, Mehmet M. 914 

Altraif, Ibrahim H. 1607 

Aluri, Krishna C. 36 

Aluvihare, Varuna 1744 

Alvarado, Edilmar A. 734, 1509 

Alvarez, Fernando 305 

Alvarez, Maria 86, 543 

Alvarez-Guaita, Anna 64 

Alvaro, Domenico 602, 621, 1654 

Alves, Erivaldo 2237 

Alves, Venancio Avancini F. 240 

Alzaabi, Mohamed 1118 

Amaddeo, Giuliana 421, 449, 1232 

Amador, Cory 1100 

Amadori, Dino 435 

Amano, Keisuke 304, 1819 

Amaral, Andreia J. 960 

Amaral, Joana D. 832 

Amarapurkar, Deepak N. 752 

Amathieu, Roland 275 

Ambade, Aditya 496, 1318, 1330, 

1351, 1958, 2127, 2198 

Ambroise, Jerome 1734 

Ambros, Victor 512 

Ameneshoa, Kelly 564 

Amer, Aliaa 1147, 1188 

Amer, Johnny 660, 1532, 1878 

Amin, Janaki 1868 

Amin, Saista 1733 

Amiot, Bruce 2 

Amiot, Xavier 2082 

Amitrano, Lucio 742 

Amiya, Takeru 181, 2131, 2140 

Ammons, Christian E. 1309 

Amorosa, Valerianna 1826 

Amoroso, Antonio 1 

Amory, Catherine 1151 

Ampuero, Javier 1134, 1863, 2171 

An, Jihyun 446 

An, Linjing 354 

Anan, Akira 385, 410 

Anand, Lovkesh 146, 262, 690, 694, 

1340, 1483, 2088 

Anand, Ravi 420 

Anand, Vijay 1791 

Ananthanarayanan, Meena 661 

Ananthavarathan, Abbesega 2047 

Anastassopoulos, Georgios 775 

Anders, Lisanne C. 2141 

Andersen, Jesper B. 677 

Anderson, Aimee 1687, 1688, 2134 

Anderson, James 20 

Anderson, Joshua 1479 

Anderson, Karl E. 2100, 2111, 2112, 

2114 

Ando, Satsuki 1474 

Andrade, Raul J. 225, 583, 595, 596, 

1863, 2171 

Andre, Remy 1072 

André, Patrice 1652 

Andreasson, Anna 158 

Andreola, Fausto 908, 1407, 1517 

Andreone, Pietro 609, 1086, 1802, 

1844, 1888 

Andreoni, Massimo 222, 1802, 1844 

Andres, Riera 202 

Andreu, Victoria 378 

Anduze-Faris, Beatrice 217 

Angeli, Paolo 148, 266, 268, 292, 

767, 775 

Angelico, Mario 203, 222 

Anguita, Juan 953 

Angus, Peter W. 106, 627, 798, 842, 

1077, 1225, 1264, 2262 

Annable, Tami 190, 835, 929, 1317, 

1349 

Annamalai, Alagappan A. 1479 

Annicchiarico, Brigida E. 432 

Annoni, Giorgio 1453 

Ansari, Aftab A. 77 

Ansari-Gilani, Kianoush 1894 

Ansolabehere, Xavier 1052 

Anstee, Quentin M. 105, 162, 2145, 

2183 

Antal-Szalmas, Peter 2090 

Anthony, Tracy 919 

Antonelli, Barbara B. 1743 

Antoniak, Silvio 81 

Antonini, Teresa Maria 1237, 1782, 

2082 

Antonucci, FrancescoPaolo 222 

Anty, Rodolphe 95, 145, 264 

Anwar, Tarique 327 

Anwer, Mohammed S. 582 

Anzai, Kazuya 383, 697 

Anzai, Keizo 2202 

Anzalone, Christopher R. 32, 2022 

Aoki, Tomoko 781, 1533, 1975 

Aoki, Yoshihiko 476, 794, 1643, 1795, 

2158 

Aono, Satoshi 26, 669, 1635, 1636, 

1661, 1809, 1982, 2130, 2136 

Aoudjehane, Lynda 30, 997, 1409 

Aouizerat, Brad 2161 

Aoyama, Tomonori 917, 1771 

Apostolova, Nadezda 1913 

Appenrodt, Beate 2077 

Applegate, Tanya L. 1083 

Apponi, Luciano 510, 2116 

Appourchaux, Kevin 998, 1601 

Apte, Udayan 650, 1767, 1774 

Aqel, Bashar A. 1038, 1124 

Arab, Juan P. 2210, 2231 

Arachchi, Niranjan J. 442 

Aracil, Carles 378 

Aragones, Julian 930 

Aragri, Marianna 222 

Arai, Jun 1302 

Arai, Kumiko 226, 917 

Arai, Kuniaki 392, 1348, 1653 

Arai, Makoto 1207 

Arai, Masahiro 1778 

Arai, Taeang 291 

Arakawa, Tomohiro 1116 

Araki, Norie 513 

Arama, Victoria 1161 

Arancibia, Juan P. 595 

Aransay, Ana M. 953 

Arase, Yasuji 1062, 1191, 2020, 2241 

Arase, Yoshitaka 314, 383 

Arauz, Oscar 1264 

Araya, Patrick 197


Arce-Cerezo, Altamira 659 

Arden, Katherine J. 1870 

Ardevol, Alba 734 

Arduino, Jean Marie 717, 729, 1867, 

1883 

Arenas, Juan I. 378, 2032 

Arendale, Evelyn 1178 

Arends, Pauline 2002, 2003 

Arendt, Bianca M. 2184, 2238, 2254 

Arendt, Karen 1111, 1155, 1164 

Arfianti, Evi 230 

Argentini, Claudio 1153 

Argo, Curtis K. 761 

Ari, Alpay 1622 

Arias, Irwin M. 20, 1909 

Arii, Shigeki 1980 

Arima, Shiho 937 

Arinaga-Hino, Teruko 304, 1819 

Ariza, Xavier 266 

Armiliato, Geyza N. 2200 

Armstrong, Matthew J. 1216 

Arnold, Frances 764 

Arnold, Hays 106, 627 

Aronsohn, Andrew I. 152, 271, 539, 

544, 553, 1229 

Aronson, Sem J. 492 

Arora, Ankur 1340 

Arora, Sanjeev 1065 

Arora, Zubin 1265 

Arosemena, Leopoldo 914 

Arput, Jean Pierre 1072 

Arranz, Jose Antonio 1509 

Arrese, Marco 595, 969, 971, 1217, 

2210, 2231 

Arriazu, Elena 1382 

Arroyo, Vicente 2069, 2071, 2087 

Arroyo-Manzano, David 745 

Arsalla, Aimal 575 

Arslanow, Anita 2253 

Arteel, Gavin E. 587, 2141, 2142 

Arterbery, Adam 1731 

Arterburn, Sarah 1045, 1049 

Artru, Florent 1269, 1782, 2222 

Arya, Aman V. 1556 

Aryafar, Hamed 913 

Asagiri, Masa 1918 

Asahina, Kinji 1354 

Asahina, Yasuhiro 164, 406, 985, 

1018, 1027, 1959 

Asahina, Yoshiro 699 

Asai, Akihiro 676 

Asai, Akira 966 

Asano, Yasukane 1420 

Asaoka, Yoshinari 461 

Asara, John 1961 

Asatryan, Armen 41, 705, 710, 719, 

723 

Asch, Steven 1048 

Ascherl, Rudi 1222 

Asghar, Aliya 724 

Asgharpour, Amon 160, 807, 905, 

1309, 2163 

Ashfaq, Mohammad 1199 

Ashouri, Besher 1100 

Asmann, Yan W. 358 

Aspichueta, Patricia 953 

Aspinall, Esther J. 1794 

Asrani, Sumeet K. 528, 1210 

Assan, Alya 1791 

Asselah, Tarik 249, 251, 714, 746, 

998, 1036, 1179, 1601, 2044, 2045 

Assene, Collins 1091 

Assimakopoulos, Stelios F. 2094, 2095 

Assis, David N. 298 

Ata, Nesrin 2051 

Ataman Hatipoglu, Cigdem 2051 

Atanasov, Georgi 1222 

Atanasov, Petar K. 1176, 1205 

Athinarayanan, Shaminie 56 

Athwal, Varinder 1364 

Atkinson, Elizabeth J. 607, 610 

Atreja, Ashish 1151 

Atsukawa, Masanori 291, 476, 1097 

Attar, Nahid 1776 

Attarwala, Husain 36 

Atwell, Thomas 293 

Aucejo, Federico N. 1271 

Aucott, Rebecca 62 

Audimoolam, Vinod K. 1588, 1763, 

2054 

Audureau, Etienne 1700 

Auger, Patrice 731 

Augustin, Salvador 742, 743 

Austin, Andrew 1480, 1524 

Austin, Patrick W. 2204 

Auth, Marcus K. 1702 

Auton, Adam 943 

Auzinger, Georg 212, 1216, 1758, 

1775 

Avagnina, Alejandra 2106 

Avani, S. 1475 

Avci, Meltem 1622 

Avila, Diana 1312 

Avila, Meera B. 1513 

Avila, Nathaniel P. 1513 

Avila, Shanika 190, 817, 834 

Avior, Yishai 679 

Avitahl-Curtis, Nicole 510, 2116 

Avitzur, Yaron 1731 

Avner, Ellis D. 1718 

Avril, Elisabeth 1796 

Ay, Nazli 444 

Ayala, Carmen 2257 

Ayer, Turgay 104, 151 

Ayers, Andrew 646, 1098 

Ayonrinde, Oyekoya T. 2185 

Ayoub, Walid S. 1479 

Aytaman, Ayse 16, 380, 438 

Aziz, Karim 264 

Aziz, Natali 123 

Azoulay, Daniel 215, 863 

Azuma, Seishin 164, 406, 1959 

Azzam, Ruba K. 1229 

Baas, Frank 637 

Baba, Hayato 1981 

Baba, Hideo 364 

Baba, Tomohisa 2119 

Bababekov, Yanik 1732 

Babatin, Mohammed A. 1607 

Bacchetti, Peter 1444 

Bach, Nancy 1095, 1185 

Bachetoni, Alessandra 983 

Back, David J. 1131 

Backstedt, David W. 1043, 1109, 

1145, 1224 

Backus, Lisa I. 93, 1786 

Bacon, Bruce 1046, 1108 

Badoe, Nina 2205 

Badra, Gamal. A. 733, 1853 

Badran, Hanaa M. 1862, 1879 

Badri, Prajakta 1066, 1084 

Badshah, Maaz B. 380, 436, 457, 

1903 

Bae, Si Hyun 452, 1615, 1882 

Baeck, Christer 961 

Baeg, Joo Yeong 389, 2113 

Baffy, Gyorgy 1469 

Bagate, François 1194 

Baggett, Sarah 1174 

Bahar, Ivet 193 

Baheti, Saurabh 600 

Bahng, Joseph 871 

Bai, Haibo 190, 514, 813, 834, 929 

Bai, Lang 1562 

Bai, Ming 754 

Bai, Shasha 1914 

Bai, Wei 468 

Baichoo, Esha 397 

Baik, Daniel 765, 766 

Baik, Soon Koo 736, 1322, 1515, 

2080 

Bailey, Lorna 1104 

Bailey, Shannon 1313 

Bailly, François 1158 

Bain, Gretchen 141 

Bain, Vince 303 

Baiocchi, Leonardo 203 

Bajaj, Jasmohan S. 52, 82, 570, 1463, 

1472, 1485, 1491, 1521, 1527 

Bakalov, Vladimir 1720 

Baker, Alastair j. 134, 258, 1691 

Baker, Talia B. 71, 838 

Bakr, Yasser M. 1821 

Bala, Shashi 109, 496, 512 

Balart, Luis A. 715, 1096 

Balasubramaniyan, Natarajan 809, 

1688, 1692 

Balasubramaniyan, Vairappan 1517 

Balba, Gayle 1034 

Balcells, Mercedes 1292 

Baldan, Angel 65 

Baldassarre, Maurizio 1493 

Balderramo, Domingo 1752 

Baldo, Vincenzo 629 

Bale, Shyam Sundhar 25 

Balestrieri, Cinzia 309, 1153 

Baliga, Prabhakar 1272 

Balk, Ethan M. 1592 

Balkrishnan, Rajesh 395 

Ball, Greg 1051, 1065, 1067 

Balladur, Pierre 1409 

Ballinger, Brad 873 

Ballot, Eric 1782 

Balteau, Sylvie 1796 

Balwani, Manisha 2111, 2112 

Bambha, Kiran 1435, 2085 

Banales, Jesus M. 21, 2171 

Bañares, Juan 1520 

Bañares, Rafael 1295, 1503, 1520 

Bandi, Sriram 1764 

Bandsma, Robert 2184 

Banerjee, Dipti 1263 

Banerjee, Rajarshi 931, 2190 

Bangma, Sander 798 

Bani, Daniele 902 

Banks, Alexander S. 881 

Bannister, Nicole 1606 

Baños, Isolina 1180 

Bansal, Meena B. 1095, 1185, 1397


Bansal, Sanjay 1704, 1712 

Bantel, Heike 2195 

Banton, Sophia 19, 2232 

Baqir, Altaf 1872 

Baquerizo, Angeles 861, 872 

Barbaro, Federico 432 

Barbati, Zachary 1090 

Barbour, April M. 730 

Barbour, Youssef 626, 1790, 1798, 

1807 

Barca, Lucia 309, 1153 

Barcia, Rita 1921 

Barclay, Murray L. 2262 

Barclay, Stephen T. 1052, 1140 

Bari, Khurram 743, 873 

Barlett, Adam 2225 

Barman, Pranab 418 

Barn, Vanessa 230 

Barnaba, Vincenzo 958 

Barnard, Trisha R. 2062 

Barnes, David S. 1265 

Barnes, Eleanor 637, 2190 

Barnes, Maisha 1146, 1169 

Barnhart, Huiman X. 225, 1922, 1923, 

1930, 1932, 1933 

Baron, Todd H. 643 

Barr, Eliav 40, 42, 210, 251, 700, 

701, 703, 712, 715 

Barr Fritcher, Emily G. 169 

Barrault, Camille 1102 

Barreiro, Pablo 438, 1180, 1881, 1908 

Barrera, Francisco 1217, 2210, 2231 

Barret, Matthieu 2188 

Barrett, Terrence 1267, 2081 

Barritt, Alfred S. 14, 533, 2093 

Barros, Raffaelle K. 2237 

Barroso, Eduardo 1256 

Barry, Jeffrey 766 

Bartels, Michael 261, 1222 

Bartenschlager, Ralf 992 

Bartolome, Sonja 9, 290 

Barton, Franca B. 286 

Barton, Ryan W. 1347 

Bartres, Concepció 982 

Barve, Shirish 114, 1304, 1312, 1331, 

1334 

Bashir, Mustafa R. 2150 

Bass, Nathan M. 1485, 1761, 2161 

Bassissi, Firas 394 

Bassit, Leda C. 1544 

Basu, Patrick 1037, 1074, 1075 

Bataller, Ramon 110, 533, 1327, 1451, 

2148 

Bate, John P. 106, 627 

Bathgate, Andrew 1216 

Batra, Sachin 1470, 1513 

Battezzati, Pier Maria 73, 634 

Batwa, Faisal 1607 

Baudesson, Camille 987 

Bauer, Christina M. 427 

Bauer, David J. 1525 

Bauer, Michael 808 

Baugé, Eric 887 

Baumert, Thomas F. 1019 

Baumgarten, Axel 1060, 1081, 1107, 

1156 

Bautista-Osorio, Eduardo 1022 

Baven-Pronk, Martine A. 302 

Bayliss, Julianne 1551, 1557, 1563 

Baytarian, Michelle 16 

Bazinet, Michel 31 

Bazzo, Maria Luiza 2075 

Beaty, Brenda 137, 1738 

Bechmann, Lars 1779, 1924 

Becker, Christina 261 

Beckerman, Rachel 877, 1534 

Beckett, Geoff 1574 

Beckford, Miguel 1961 

Becquart, Jérôme 1409 

Bedard, Patricia W. 1770 

Bedossa, Pierre 105, 162, 480, 887, 

998, 1601, 2145, 2148, 2164 

Beets, Greet 39 

Behari, Jaideep 1496, 1531 

Behling, Cynthia A. 157, 159 

Behrns, Kevin E. 177 

Beier, Juliane I. 2141 

Beigelman, Leo 993 

Beilby, John 2099 

Beilin, Lawrence 2185 

Beinhardt, Sandra 297, 732, 1070, 

1092, 1137 

Beiser, David G. 522 

Bejarano-Fernandez, Eloy 333 

Bekki, Shigemune 408, 783, 1600 

Béland, Kathie 305 

Belghiti, Jacques 417 

Bell, Chaim M. 350 

Bell, David 424 

Bell, Lauren N. 161 

Bell, Phillip D. 962, 2137 

Bell, Sally 442, 483 

Belle, Steven H. 121, 1598 

Bellelli, Giuseppe 1453 

Bellentani, Stefano 2164 

Belli, Dominique 1735 

Belli, Luca S. 571, 1751 

Bellini, Giulia 1837 

Belloni, Laura 958 

Bellos, Damien 2132 

Belloula, Djamel 554 

Belperio, Pamela S. 93, 1786 

Belt, Patricia H. 157, 159 

Bemeur, Chantal 51 

Ben Ari, Ziv 94, 1032, 1685 

Ben Mosbah, Ismail 215 

Benbow, Jennifer H. 1389 

Bendersky, Noelle 373 

Benedetti, Antonio 21 

Benetti, Gianpiero 148 

Benfatto, Salvatore 1127, 1654 

Benidir, Andreanne 1713 

Beninati, Concetta 1127, 1654 

Benitez, Laura 1881 

Benítez, Carlos E. 1217, 2210, 2231 

Benítez-Gutiérrez, Laura M. 1180 

Benito, Yolanda 1503 

Benjamin, Jaya 1328, 1340, 2088, 

2203 

Benjaram, Sindhuri 420 

Benkert, Pascal 1884 

Benlloch, Salvador 1230, 1251 

Bennai, Yacia 206 

Bennett, Michael 248, 250, 1039 

Benson, Ariel 1878 

Benyashvili, Tamara 1172 

Benzing, Christian 1222 

Beppu, Toru 364 

Berardi, Sonia 1751 

Berenguer, Marina 1084, 1230, 1251 

Bereshchenko, Oxana 1963 

Beret, Antoine 394 

Beretta, Laura 170, 1424, 2151 

Berg, Christoph P. 76, 1055, 1183 

Berg, Thomas 37, 245, 249, 252, 261, 

276, 308, 1058, 1594, 1638, 1678, 

1861, 2003, 2004, 2012 

Bergeat, Damien 487 

Berger, Calise K. 169 

Bergin, Colm J. 1103 

Bergquist, Annika 76 

Berhane, Sarah 344, 851 

Berk, Paul D. 972 

Berkane, Saadi 1621 

Berkes, Jamie 1248 

Berkowski, Caterina 980, 1055, 1559 

Bermejo, Javier 1503 

Bernabucci, Veronica 1751 

Bernal, William 212, 740, 1775 

Bernard, Denis 1239 

Bernard-Chabert, Brigitte 1072 

Bernardi, Mauro 1493, 2087 

Bernstein, David 726, 1039, 1051, 

1065, 1067, 1086 

Berntsen, Natalie L. 819 

Bernuzzi, Francesca 602, 621 

Berres, Marie-Luise 1364 

Berry, Kristin 49, 1211 

Bertelli, Cristina 393 

Bertero, Alessandro 258 

Bertinetto, Francesca E. 1 

Bertoletti, Antonio 167 

Besch, Camille 3, 95 

Beschorner, Niklas 1629 

Besisik, Fatih 478 

Bessho, Ryosuke 1981 

Bessone, Fernando 225, 595 

Best, Jan 1779 

Beste, Lauren A. 128, 540 

Besur, Siddesh 17 

Betrapally, Naga 1463, 1527 

Bettencourt, Ricki 913, 1431, 2201 

Bettinger, Dominik 1803 

Beudeker, Boris 1007 

Beuers, Ulrich 74, 76, 317, 492, 604, 

623, 637, 827, 1995, 2152 

Beumont, Maria 39 

Bevans-Fonti, Shannon 1422 

Beyer, Jill 705 

Beysen, Carine 1425 

Bezerra, Jorge A. 133, 676 

Bhadoria, Ajeet S. 1268, 1328, 2203 

Bhalla, Ashish 259 

Bhamidimarri, Kalyan R. 200, 1054, 

1148, 1206, 1587, 2256 

Bhandari, Bal R. 248, 250 

Bhanji, Rahima A. 303, 1481 

Bhardwaj, Ankit 1328, 1340, 1659, 

2203 

Bhat, Sathesh P. 957, 1938 

Bhatia, Vikram 2070, 2088 

Bhatnagar, Raj K. 1659 

Bhatt, Archana 913, 1431 

Bhattacharya, Dipankar 1392 

Bhattacharya, Renuka 1221 

Bhoori, Sherrie 1751 

Bhore, Rafia 706, 716, 726 

Bhuket, Taft 7, 237, 1252, 2159 

Bhushan, Bharat 1767 

Bianchi, Ilaria 602


Bibert, Stephanie 221 

Bichoupan, Kian 1095, 1151, 1185, 

1202 

Bider-Canfield, Zoe 1135, 1171 

Bieghs, Veerle 100 

Bigaeva, Emilia 1434 

Biggins, Scott W. 82, 570, 2085 

Bihari, Chhagan 327, 690, 694, 1483, 

2078 

Bilasy, Shymaa E. 858, 2215 

Biliotti, Elisa 222, 983 

Billaud, Eric 568, 1093 

Billiar, Timothy R. 656 

Billingsley, Kevin 481 

Bilodeau, Marc 179, 204 

Bilzer, Manfred 1170, 1200 

Binder, Christoph J. 922 

Bindmann, Julia 98 

Biolato, Marco 432 

Birerdinc, Aybike 2205 

Birnbaum, Morris J. 65 

Bisch, Grégoire 1409 

Biselli, Maurizio 1493 

Bishnu, Saptarshi 750 

Bissell, D Montgomery 2111, 2112 

Bissonnette, Julien 772, 2148 

Bizzotto, Paola 775 

Bjoernetroe, Tonje 819 

Bjornsson, Einar 225 

Blach, Sarah 1818, 1842 

Black, Sylvester 874, 1260, 1280 

Blacklidge, Judith 200 

Blackman, Brett R. 175, 664, 915, 

1916 

Blanc, Etienne 30 

Blanc, Jean-Frédéric 132, 450, 851 

Blanc, Pierluigi 1802, 1844 

Blanco, Albert 1509 

Blandino, Giovanni 958 

Blank, Emma E. 507 

Blas-García, Ana 1913 

Blasco-Perrin, Hélène 2082 

Blasi, Maria 1021, 1674 

Blatt, Lawrence M. 993 

Bloch, Mark 210 

Block, Timothy 371 

Bloomer, Joseph R. 2111, 2112 

Blumberg, Richard S. 819, 833 

Blumenthal, Antje 1362 

Bobardt, Michael 2035 

Boberg, Kirsten M. 76, 602 

Bochud, Pierre-Yves 221 

Bock, Izona 1874 

Bockamp, Ernesto 183 

Bode, Konrad A. 618 

Boeck, Christina 444 

Boehlig, Albrecht 276 

Boehm, Stephan 245, 276, 1594, 1638 

Boehme, Shannon M. 885 

Boeker, Klaus H. 1861 

Boekschoten, Mark V. 1924 

Boemio, Adriana 1837 

Boesecke, Christoph 1060 

Boettler, Tobias 2195 

Boffelli, Silvia 1493 

Bohorquez, Humberto E. 1242 

Boin, Ilka F. 469 

Boix, Loreto 65 

Bojito-Marrero, Lizza 295, 1181 

Bolcas, Paige 823 

Boldt, Andreas 261 

Boleslawski, Emmanuel 487, 1269 

Bolier, Ruth 827 

Bollipo, Steven J. 106, 627 

Bolognesi, Massimo 292, 767 

Bombonato, Giancarlo 292, 775 

Bond, Geoffrey 763 

Bondy, Carolyn 1720 

Bonello, John 2251 

Bonkovsky, Herbert L. 597, 1023, 1923, 

2111, 2112 

Bonnefont-Rousselot, Dominique 786 

Bono, Patrizia 1747 

Bonomo, Lorenzo 432 

Boodram, Basmattee 1854 

Boone, Pamela 1229 

Boonsirichan, Rattana 1631, 1662 

Boonstra, Andre 1007, 1196, 1585, 

1586, 1590, 1637, 2122 

Boonstra, Kirsten 74, 623 

Boor, Peter 961 

Booty, Jordan 521 

Bopp, Tobias 1353 

Borad, Mitesh J. 396, 1965, 1966 

Borchardt, Hannes 1299, 1385 

Borg, Brian B. 1178 

Borgeaud, Nathalie 665 

Borghesan, Michela 1445 

Borgia, Guglielmo 1802, 1844 

Borgne, Anne 1796 

Borok, Zea 1945 

Borralho, Pedro M. 176, 897, 960 

Borralho Nunes, Paula 940 

Borst, Andrew 1316 

Bortoluzzi, Ilaria 1270 

Borude, Prachi C. 1774 

Borzio, Franco 355 

Boscarino, Joseph A. 15, 525, 1578, 

1789, 1799, 1800 

Bosch, Jaime 326, 737, 739, 745, 746, 

1418, 1518, 2239 

Bosma, Piter J. 492, 827 

Bosman, Dean 724 

Bosoi, Cristina R. 51, 1520, 1523 

Bossi, Krista 2166 

Boström, Adrian 519 

Bota, Simona 270, 475, 732, 755, 771 

Botta-Fridlund, Danielle 3, 95, 206 

Bottai, Matteo 116 

Bottari, Antonio 288 

Bottero, Julie 1677 

Botwin, Gregory J. 724, 1129 

Bouattour, Mohamed 417, 480 

Boudjema, Karim 487 

Boudon, Marc 1237 

Boueyre, Estelle 1194 

Boulanger, Chantal 887, 2148 

Boulter, Luke G. 678 

Bourcier, Valérie 362, 1808, 1847 

Bourgeois, Stefan 797, 1051, 1091, 

1842 

Bourhis, Francois 1072 

Bourlière, Marc 206, 249, 394, 777, 

1442, 1860, 2082 

Boursier, Jerome 117, 1439, 2188 

Bouvier, Antoine 263 

Bouvier-Alias, Magali 1123 

Bouyssou, Caroline 132 

Bovitz, Tanya 1867, 1883 

Bowden, Scott 1557, 1563, 1566 

Bowe, Andrea 687 

Bowen, David 1243 

Bowen, William C. 670, 1979 

Bowlus, Christopher L. 76, 77, 602, 

606, 609, 613, 616, 624, 630, 639, 

1034 

Bowring, Mary G. 1749 

Bowyer, Teresa 1588, 2053 

Box, Terry D. 2257 

Boyd, Anders 1676, 1677 

Boyer, James L. 23, 298, 805, 829 

Boyer, Nathalie 206, 998, 1601, 2044, 

2045 

Boyer, Thomas D. 278, 1459 

Boyett, Sherry L. 905, 2235, 2249 

Bozdayi, A.Mithat 2033 

Bozorgzadeh, Adel 845 

Bradley, J. Andrew 258 

Brady, Joanne E. 1792, 1839 

Brahm, Javier 595 

Brahmania, Mayur 542, 1556 

Braid-Forbes, Mary Jo 531 

Brain, John G. 611 

Brainard, Diana M. 38, 91, 96, 205, 

219, 247, 249, 713, 718, 746, 1034, 

1041, 1049, 1080, 1094, 1120, 

1128, 1442, 1860 

Brancatelli, Santa 1898 

Branch, Andrea D. 29, 229, 988, 1095, 

1112, 1151, 1185, 1202, 1289, 

1397, 1742 

Branco, Fernanda 469 

Brandan, Enrique 969 

Brandl, Katharina 59 

Brandman, Danielle 414, 1273 

Brandon-Warner, Elizabeth 1389 

Brantly, Mark 138 

Braspenning, Joris 331 

Bratthauer, Gary 891, 956 

Bratton, Charles F. 1272 

Bräu, Norbert 205, 380, 438, 457 

Braun, Felix 76 

Braun, Marius 875 

Breckenridge, David 1359 

Breen, Gerome 2105 

Brejda, John 725 

Bremer, Birgit 1572, 1597, 2026 

Bremer, Corinna M. 2026 

Brenner, David A. 59, 186, 187, 189, 

890, 1415, 1425, 1431 

Breuer, Monika 46 

Bricault, Ivan 421, 449 

Bridge, Robert S. 1969, 2139 

Bridges, Brian 352, 1291, 1814, 1815 

Bridges, John F. 1727, 1729 

Bridle, Kim 818, 975 

Brigham, Benjamin S. 36 

Brigstock, David 192, 1356, 1404 

Brimacombe, Michael B. 1774 

Briones-Orta, Marco A. 1978, 2143 

Briot, Charline 264 

Brisac, Cynthia 25, 991, 1015, 1630, 

1644 

Britton, Laurence J. 818, 975 

Brivio, Simone 1950 

Brixko, Christian 1842 

Brock, Guy N. 2233 

Brodsky, Allison 1864, 1880 

Bronich, Tatiana 589 

Bronk, Steven 825, 895


Bronowicki, Jean-Pierre 206, 2082 

Brookes, Steve 424 

Brookmeyer, Ron 1068 

Brooks, Alexandria 536 

Brooks, Louis 1805 

Brosch, Mario 617 

Broschewitz, Johannes 1222 

Brouillet, Arthur 693, 1391 

Brouwer, Linda 1416 

Brouwer, Willem Pieter 348, 1568, 

1585, 1586, 1590, 1637, 2001, 2003 

Brown, Ashley S. 1826, 2003, 2047 

Brown, Bob D. 510, 2116 

Brown, Deborah D. 712, 717 

Brown, Helen 44 

Brown, Kimberly Ann 307, 1084, 1792 

Brown, Maxwell A. 1754 

Brown, Robert 1037, 1049, 1074, 

1075 

Brown, Robert E. 905 

Brown, Robert S. 217, 272, 369, 1084, 

1454 

Bruce, David S. 1242 

Bruce, Matthew J. 1113, 1588, 1704, 

1744, 2053, 2054 

Bruce, Michael 1790, 1798 

Bruce, Robert 1874 

Bruckbauer, Antje 933 

Bruden, Dana J. 1790, 1798, 1807 

Bruggmann, Philip 1818 

Bruha, Radan 1487 

Bruix, Jordi 65, 378, 860 

Brumme, Chanson J. 1832 

Brun, Sonia 394 

Brunetto, Maurizia R. 1802, 1844 

Brunner, Jim 1839 

Bruno, David A. 1274 

Bruno, Raffaele 1802, 1844 

Bruns, Tony 73 

Brunt, Elizabeth M. 157, 159, 2150, 

2220 

Bruscella, Patrice 987 

Brusch, Lutz 617 

Brusilovskaya, Ksenia 1525 

Brusquant, David 417 

Bryan, William E. 1142 

Bucala, Richard 298 

Buchanan, Charlotte E. 1480 

Buchholz, Frank 1629 

Bucsics, Theresa 270, 771, 1204 

Budoff, Matthew 1488 

Buechler, MarkusW. 1227 

Buggisch, Peter 1078, 1170, 1176, 

1179, 1200, 1205, 1559, 1861, 2257 

Bugianesi, Elisabetta 973, 2164, 2209 

Buist-Homan, Manon 591 

Buitrago, Laura E. 2128 

Bukong, Terence N. 1413, 2198 

Bulkow, Lisa 125 

Bull, Laura 78 

Bulut, Cemal 2051 

Bunchorntavakul, Chalermrat 1631, 

1662 

Bunniran, Suvapun 1050 

Bunraksa, Worawan 2089 

Burchill, Matthew A. 28 

Bureau, Christophe 145, 263, 264, 

744, 2082 

Burgess, Gary 798 

Burke, Ann E. 1452 

Burks, Deborah J. 691 

Burns, Justin M. 50, 53 

Burra, Patrizia 268, 351, 1270, 1889 

Burrin, Douglas 194, 1689 

Busch, Heiner W. 1060, 1081, 1156 

Bushau, Adrienne M. 2141 

Bushyhead, Daniel W. 1218 

Busuttil, Ronald W. 1283, 2138 

Buti, Maria 241, 243, 1021, 1179, 

1674, 2003, 2012, 2014, 2032, 

2037, 2059 

Butt, Adeel 1488 

Butt, Mohammed T. 1147 

Butte, Atul 497, 500 

Butterfield, David 673 

Butterton, Joan R. 701, 717, 725, 730, 

731 

Butterworth, Jacqueline 652 

Buzzigoli, Emma 2209 

Byrne, Sean 1041 

Byun, Kwan Soo 391, 405, 1504, 

1897, 2017 

Bzeizi, Khalid I. 1607 

Bzowej, Natalie H. 1242 

Caballeria, Juan 1307, 1327, 2148 

Caballeria, Llorenç 73, 629, 634 

Cabarrou, Pauline 263 

Cabello-Verrugio, Claudio 969 

Cabellos, Laia 253 

Cabibbo, Giuseppe 355 

Cable, Rebecca 2204 

Cabrera, Daniel 969, 971 

Caccamo, Gaia 288, 1476 

Caccamo, Lucio 1743 

Cacciarelli, Thomas V. 1753 

Cacciola, Irene 288, 1127, 1476 

Cachero, Alba 742 

Cacoub, Patrice 2082 

Cadamuro, Massimiliano 1950 

Cadranel, Jean François D. 264, 554, 

2082 

Cai, Dachuan 991, 1015, 1630, 1644 

Cai, Hongwei 468 

Cai, Hou Ming 197 

Cai, Luke 1284 

Cai, Shi-Ying 805, 2179 

Cai, Yan 110, 903 

Caiazzo, Robert 2222 

Caines, Allyce 532, 559 

Caire, Thure 526, 1240 

Cakaloglu, Yilmaz 558, 2001 

Cakmak, Erol 2219 

Calderaro, Julien 421, 449, 693, 1232 

Caldwell, Stephen H. 13, 105, 162, 

737, 761, 1428, 2145, 2239 

Cales, Paul 105, 117, 162, 741, 2145, 

2188 

Caligiuri, Alessandra 1410 

Calinas, Filipe 1859 

Caliri, Kimberley 2257, 2259, 2260 

Caliskan Kartal, Aysun 2033 

Calleja, José Luís 746, 1511, 2012, 

2032, 2171 

Calmus, Yvon 30, 997, 1239, 1409 

Caloggero, Simona 288 

Calvisi, Diego 1934, 1948 

Cam, Margaret 218 

Camacho, Jocelyn A. 1870 

Cambe, Joy 1870 

Cambula, Linda 974 

Cameron, Andrew M. 1255, 1727, 

1729, 1749 

Campana, Lara 62 

Campanale, Mariachiara 432 

Campbell, Andrew L. 705 

Campbell, Catherine M. 818 

Campbell, Deanna J. 1993 

Campbell, Jean S. 1395 

Campos, Frederico F. 2189 

Campos, Laura 1991 

Campos, Priscila B. 2221 

Camprecios, Genis 253, 254 

Campsen, Jeffrey 200 

Canbay, Ali 1779, 1924, 1978, 2143 

Canchola, Jesse A. 1597 

Cancino, Alejandra 1217 

Candotti, Fabio 1717 

Canini, Irene 1153 

Canini, Laetitia 982 

Canizaro, Leticia 722 

Cannon, Ryan 1069 

Canva, Valerie 3, 2222 

Cao, Chuanhui 1939 

Cao, Furong 1541, 1579, 1617, 1773 

Cao, Jiawei 1998 

Cao, Sheng 1315, 1355, 1406 

Cao, Xiaoming 169 

Cao, Ying 2066 

Cao, Yirong 131 

Cao, Yumei 1048 

Cao, ZhuJun 1611 

Cao, Zongxian 1374 

Caparosa, Susan 87 

Capocelli, Kelley E. 1709, 1715 

Capoluongo, Nicolina 1837 

Cappellini, Sara 309 

Cappoli, Andrea 983 

Cappon, Andrea 973 

Caprio, Nunzio 1543 

Caraceni, Paolo 1493, 1751, 1802, 

1844 

Carbone, Marco 73, 155, 601 

Carbonell, Nicolas 263, 264 

Carcassi, Carlo 309 

Cardenas, Andres 2084 

Cardenas, Victor M. 1857 

Cardillo Marricco, Nadia 725, 731 

Cardoso, Alberto 473 

Cardoso, Filipe S. 1759 

Cardoso, Helder 1540 

Cardoso de Brito, Miguel 258 

Cardozo, Karina H. 48 

Carey, Elizabeth J. 106, 605, 627, 

1468 

Carey, Ivana 1098, 1113, 1117, 1132, 

1173, 1571, 1588, 1626, 1704, 

1744, 2003, 2053, 2054 

Caridade, Marta 897, 940 

Carito, Brenda 36 

Carl, Daniel 1521 

Carli, Fabrizia 2209 

Carlson, Rolf I. 256, 1962 

Carlton-Smith, Charles 1015 

Carmody, Ian C. 1242 

Carmona, Damien 1102 

Caro, Luzelena 707, 725, 731 

Carpentier, Arnaud 30 

Carpino, Guido 817 

Carr, Brian I. 437 

Carr, David 260, 753


Carr, Kathy 1433 

Carr, Rotonya M. 1314 

Carrat, Fabrice 206 

Carrera, Enrique 595 

Carriero, Damaris C. 1112 

Carrilho, Flair J. 48, 240, 469, 1033, 

1544, 2221 

Carter, Jeffrey D. 1141 

Carvalho, Armando 473 

Carvalho, Catarina C. 897 

Carvalho, Tânia 940 

Carvalho, Valdemir M. 48 

Casadei Gardini, Andrea 435 

Casado, Marta 2032 

Casale, Michele 309, 1153 

Casalino, Paolo 222 

Casals, Gregori 659 

Casanovas, Teresa 2032 

Cascinu, Stefanu 435 

Cascorbi, Ingolf 225 

Casey, Carol A. 1310, 1969, 2139 

Casey, Stephen 798 

Casillas, Rosario 1021, 1674 

Casillas-Ramírez, Araní 342 

Casingal, Vincent P. 17 

Cassader, Maurizio 2209 

Cassagnol, David 1112 

Cassiman, David 171 

Cassinotto, Christophe 450 

Cassio, Doris 816 

Cast, Ashley E. 680 

Castaing, Denis X. 487, 1237 

Castaño, Gustavo O. 2217 

Castellani, Paul 394 

Castellote, Jose 742 

Castelnau, Corinne 2044, 2045 

Castera, Laurent 417, 480 

Castiella, Agustin 583 

Castiello, Filomena 431 

Castillo, Alvaro E. 481 

Castillo, Eliana 1576 

Castro, Azucena 620, 2244 

Castro, Marcelo 2106 

Castro, Rui E. 176, 832, 897, 940, 960 

Castro Filho, Elio C. 84 

Castroagudin, Javier F. 378 

Casu, Stefania 309, 1153 

Casulleras, Mireia 2087 

Casuscelli di Tocco, Francesca 1898 

Catalano, Donna 496, 512 

Catalina, Maria Vega 1503 

Catalli, Lisa 1825 

Catone, Stefania 1153 

Cattral, Mark S. 11 

Caturelli, Eugenio 355 

Causse, Xavier 1072, 2082 

Cavalcanti, Marta 762 

Cavallin, Marta 148 

Cave, Matthew C. 666, 1303, 2141, 

2233 

Cavenago, Margherita 1743 

Caviglia, Gian Paolo 1740 

Cazanave, Sophie C. 160, 905 

Cazier, Alain 554 

Cazzagon, Nora 73, 299, 629 

Cebotarescu, Valentin 31 

Ceccherini-Silberstein, Francesca 222 

Cedars, Marcelle I. 2252 

Cehic, Damir G. 1021 

Celik, Neslihan 763 

Cellai, Ilaria 882, 902 

Ceni, Elisabetta 949, 1963 

Cento, Valeria 222 

Cereser, Biancastella 101 

Cerny, Andreas 221, 1884 

Cerocchi, Orlando 1099, 1556 

Cervoni, Jean Paul 264, 1036, 1187 

Cesana, Giancarlo 571 

Cesario, Valentina 432 

Cha, Seung-Kuy 1401 

Chaabna, Karima 1829 

Chagas, Aline 48, 469 

Chagneau-Derrode, Carine 2082 

Chahal, Barinder 1413 

Chainuvati, Siwaporn 467 

Chaiprasert, Amnart 2089 

Chait, Alan 923 

Chaiteerakij, Roongruedee 127, 360, 

397, 416, 1497, 1965, 1966 

Chakrabari, Subrata 1246 

Chakraborty, Shweta 1814, 1815 

Chakraborty, Souvik 197 

Chalasani, Naga P. 225, 236, 239, 

436, 607, 624, 948, 1316, 1332, 

1903, 1922, 1923, 1926, 1930, 

1933, 2154, 2166, 2175, 2220 

Challies, Tracy L. 2160 

Chamberland, Robin 1535 

Champion, Mia D. 1965 

Chamulitrat, Walee 1411 

Chan, Albert 70, 1746 

Chan, Anthony W. 238, 2181 

Chan, Che-Chang 965 

Chan, Henry Lik-Yuen 235, 238, 241, 

349, 1120, 1548, 1557, 1563, 1612, 

1651, 2014, 2025, 2181 

Chan, Kelvin K. 350 

Chan, Sarah Y. 307 

Chan, See-Ching 70, 1746, 2000 

Chan, Stephen L. 344 

Chandhok, Gursimran 684, 2103 

Chandra, Partha K. 501, 506, 1096 

Chandrasekar, Edwin 579 

Chandrasekaran, Suki 2091 

Chandrashekaran, Varun 962 

Chang, Albert 471 

Chang, Binxia 903 

Chang, Chao-Hsuan 619 

Chang, Charissa Y. 1095, 1185 

Chang, Chi-Yang 1547 

Chang, Chia-Lin 1593, 1604 

Chang, Christine Y. 123, 359, 388, 

1182, 1189, 1806, 1816, 1848, 2009 

Chang, Chung-Chou 1488 

Chang, David C. 1732 

Chang, Ellen T. 1668 

Chang, Hye Young 1399 

Chang, Jason 451 

Chang, Kuo-Chin 1873, 1942 

Chang, Kyong-Mi 1001 

Chang, Mei-Hwei 1701, 1705 

Chang, Na 1396 

Chang, Paul 765, 766 

Chang, Sanders 1202 

Chang, Ting-Tsung 1554 

Chang, U Im 1615 

Chang, Will 2247 

Chang, Ya-Ju 654 

Chang, Yung-Sheng 1639 

Chang, Young 474, 479, 1610, 2016 

Channick, Richard 9 

Chao, Xiaojuan 598 

Chapkin, Robert 56 

Chapman, Roger W. 74, 616, 622, 

624, 637 

Chapman, William C. 704 

Charatcharoenwitthaya, Phunchai 467 

Charbek, Edward 1535 

Charier, Florian 95 

Charisse, Klaus B. 36 

Charles, Edgar 712 

Charles, Hoppel 518, 651, 667 

Charlot, Hélène 1102 

Charlton, Carmen L. 1576 

Charlton, Michael R. 96, 616, 746, 

1045, 1049, 1130, 1226, 1359, 

1442, 2149 

Charnaux, Nathalie 362 

Charuworn, Prista 241, 2025 

Chasan, Rachel 1870 

Chascsa, David M. 1498 

Chatterjee, Saswata 750 

Chatterjee, Saurabh 962, 2133, 2137 

Chatterji, Udayan 2035 

Chattieng, Piyanat 360 

Chaturvedi, Prasoon 36 

Chau, Gar-Yang 439 

Chaubey, Pururawa M. 806 

Chaudhry, Asad A. 1872 

Chaudhry, Hunana 1864, 1880 

Chaudhry, Sulemon 2129 

Chauhan, Ranjit 1641 

Chaung, Kevin T. 207, 368, 1561 

Chava, Srinivas 501, 506, 1096 

Chavan, Shailesh 200 

Chavez, Deborah 32, 2022 

Chavin, Kenneth D. 1272 

Chawla, Yogesh K. 214 

Chayama, Kazuaki 472, 663, 707, 

986, 1056, 1150, 1627, 1645, 1649, 

1670, 1679, 1681, 1858, 1893, 

1895, 1944, 2120, 2156, 2199, 2202 

Chazouillères, Olivier 76, 95, 624, 642 

Che, Li 1934 

Cheetham, T. Craig 87, 1135, 1171, 

1788 

Chekuri, Sweta 1095, 1202 

Chelis, Leonidas 1900 

Chelvaratnam, Uthayanan 1264 

Chemaitelly, Hiam 1829 

Chemello, Liliana 1802, 1844 

Chemnitz, Jan 1629 

Chen, Anping 911, 1361, 1365 

Chen, Bin 497, 500 

Chen, Chi-Hua 1431 

Chen, Chi-Ling 1996 

Chen, Chi-Yi 1996 

Chen, Chia-Lin 1945 

Chen, Chien-Hung 1873, 2010 

Chen, Chien-Jen 207, 1561, 1593, 

1604, 1828 

Chen, Chien-Yu 1945 

Chen, Ding-Shinn 2027 

Chen, Dongyu 510, 2116 

Chen, Erluo 700 

Chen, Feng 680 

Chen, Guang 1656, 1683, 1684 

Chen, Gui-Hua 419, 440 

Chen, Guofeng 1536, 1537, 2008 

Chen, Hank H. 1450


Chen, Hong-Song 1973 

Chen, Huan 353, 375 

Chen, Huey-Ling 1701, 1705 

Chen, Jian 130, 682, 2118 

Chen, Jianhong 1595 

Chen, Jiegen 1327 

Chen, Jing 1536, 1537, 2008 

Chen, Jiun-Sheng 130, 682 

Chen, Li 192, 1356, 1404 

Chen, Liang 1998 

Chen, Limin 981 

Chen, Long 782 

Chen, Michael 1986 

Chen, Ming-Yao 1996 

Chen, Minjun 596 

Chen, Minshan 851 

Chen, Pei-Jer 1593, 1604, 2027 

Chen, Peng 111, 1319 

Chen, Ping-Hsien 738, 751, 757, 759 

Chen, Po-Hung 1214, 1749 

Chen, Ruju 192, 1356 

Chen, Shi-Jun 2023 

Chen, Shiyao 131 

Chen, Steve 2175 

Chen, Tao 1660 

Chen, Tianyan 1541, 1579, 1617, 

1773 

Chen, Ting-Yi 380, 438, 457 

Chen, Tsung-Ming 1996 

Chen, Vincent L. 367, 387, 428, 459, 

1668, 2009 

Chen, Wei 454 

Chen, Wei-Yang 1304 

Chen, Weina 686 

Chen, Wen-Chi 738 

Chen, Xiangmei 1634 

Chen, Xin 1934, 1948 

Chen, Xinyue 2048, 2061 

Chen, Yi-Cheng 1552 

Chen, Yi-Ju 345, 1589 

Chen, Ying 1693 

Chen, Ying-chun 619 

Chen, Yonglin 979, 2179 

Chen, Yu 1382, 1940 

Chen, Zhaochun 1646 

Chen, Zhi-wei 1044 

Chen, Zishuo 904 

Cheng, Frank 285 

Cheng, Jin 1634 

Cheng, Jin-Shiung 738 

Cheng, Jun 2013 

Cheng, Li-sha 1383 

Cheng, Linling 1430 

Cheng, Rex 1221 

Cheng, Stephen 1169 

Cheng, Sunny Lihua 655 

Cheng, Wendy 2262 

Cheng, Yang 167 

Cheon, Gab Jin 277, 484, 1515 

Cheon, HyeonJoo 1025 

Cheong, Jae Youn 379, 1567 

Cheraitia, Salima 1621 

Chéret, Antoine 568, 1093 

Cherk, Erika 549 

Chermak, Faiza 450, 2181 

Cherniack, Andrew D. 129 

Cheroni, Cristina 2011 

Cherqui, Daniel 449, 487, 1237 

Chessa, Luchino 309, 1153, 1802, 

1844 

Cheung, Angela C. 73, 634 

Cheung, Cindy K. 1746 

Cheung, Ka-Shing 126 

Cheung, Michael 1261, 1276 

Cheung, Michelle C. 1173 

Cheung, Tan To 70, 1746 

Chevaliez, Stéphane 999, 1015, 1123, 

1700, 1797, 1833 

Chhatwal, Jagpreet 104, 151 

Chi, Aileen 1560 

Chi, Benjamin 1575 

Chi, Heng 1998, 2002, 2003, 2012 

Chi, Xiumei 1149, 1193, 1603, 1869 

Chi-Cervera, Luis A. 1471 

Chiang, John 885 

Chiang, Kevin 87, 1788 

Chiaradia, Mélanie 421, 449 

Chiaramonte, Maria 355 

Chiba, Tsutomu 1989 

Chiba, Yousuke 1369 

Chibaudel, Benoist 417 

Chida, Takeshi 1393 

Chie, Wei-Chu 1701 

Chien, Rong-Nan 1552, 1996 

Chikada, Hiromi 697, 1369 

Chikhi, Yazid 1621 

Childs, Kate 1113 

Childs, Kate E. 646, 1098, 1117, 1132 

Chim, Angel ML 238, 1548, 2181 

Ching, Helena 2185 

Chinnasamy, Thirunavukkarasu 340 

Chiodo, Letizia 165 

Chiou, Yu-Wei 1639 

Chipumuro, Edmond 2116 

Chirapongsathorn, Sakkarin 281, 282, 

2089 

Cho, Eun Ju 242, 400, 447, 470, 474, 

479, 1610, 1907, 2016, 2042, 2055 

Cho, Eun-Suk 464 

Cho, Hyeki 242, 479, 2016 

Cho, Hyo Jung 379, 1567 

Cho, Hyosun 1001 

Cho, Jai Young 857, 1902 

Cho, Ju-Yeon 1415 

Cho, Junhyeon 389 

Cho, Kyung Joo 504 

Cho, Mee-Yon 1322 

Cho, Mong 876, 1542, 2028 

Cho, Soo Yeon 2056 

Cho, Sung Won 379, 1567, 2056 

Cho, Yong Kyun 924, 941, 951, 1624, 

1760, 1992, 2110, 2218, 2243 

Cho, Young Youn 242, 400, 447, 474, 

479, 1610, 1907, 2016 

Cho, Yuri 242, 400, 447, 474, 479, 

1610, 2016, 2042, 2055 

Chodavarapu, Krishna 713 

Chodavarapu, Ramakrishna K. 1168 

Choi, Angela O. 725 

Choi, Cheol Woong 876 

Choi, Dae Hee 1515 

Choi, Duck Joo 391, 1896 

Choi, Gina 122 

Choi, Hanlim 857 

Choi, Hwa Young 2113 

Choi, Jong Young 452, 1615, 1882 

Choi, Jonggi 127, 360, 381, 397 

Choi, Jung Eun 502, 1419 

Choi, Moon Seok 422, 423, 485, 

1591, 1960, 2049 

Choi, Myunghan 1043, 1109, 1145, 

1224, 1241, 1262 

Choi, Paul C. 238 

Choi, Sang Il 377 

Choi, Steve S. 80, 1142 

Choi, Won-Mook 1358, 1907 

Choi, Yong-Hun 403, 426, 749 

Choi, Yoo-Shin 1278 

Choi, Youkyung 2126 

Choi, YoungRok 857, 1902 

Chok, Kenneth S. 70, 1746, 2000 

Chokshi, Shilpa 163, 599, 2143 

Cholankeril, George 372 

Chong, Hui Heng 1686 

Chong, Yutian 120 

Chopda, Girish R. 2116 

Chopra, Kapil B. 607, 648, 1754 

Chotiyaputta, Watcharasak 467 

Chou, Hsin I 1364, 1392 

Chou, Wen-Wen 2264 

Choudhary, Manish C. 1005 

Choudhary, Narendra S. 2245 

Choudhury, Ashok K. 103, 1344, 2070, 

2078 

Chousterman, Michel 567 

Chow, Eric 1727, 1729 

Chowdhary, Vivek K. 99 

Chowdhury, Abhijit 750 

Christensen, Peer B. 792 

Christensen, Stefan 1052, 1060, 1081, 

1156, 1861 

Chromy, David 561, 1204 

Chu, Andrew 193, 197 

Chu, Chia-Ming 1552 

Chu, Po-sung 181, 569, 2131, 2140, 

2234 

Chu, Winnie C. 235 

Chua, Cui Wen 1812 

Chua, Mei-Sze 497, 500 

Chuang, Wan-Long 91, 241, 367, 398, 

459, 1080, 1105, 1120, 1168, 1828, 

1996, 2014, 2025, 2264 

Chuang, Ya-Hui 619 

Chuaypen, Natthaya 2036, 2063 

Chui, Celia B. 1832 

Chun, Hoon Jai 1504 

Chun, Youngjae 216 

Chung, Jung Wha 389 

Chung, Kyu Sik 346, 1542, 1558 

Chung, Randy 357 

Chung, Raymond T. 25, 154, 704, 778, 

991, 1015, 1094, 1630, 1644, 1768 

Chung, Sook In 504 

Chung, Sookja K. 1961 

Chung, Woo Jin 2038 

Chung, Young-Hwa 446, 448 

Churchill, Norma D. 1641 

Ciaccio, Antonio 571, 1453 

Ciancio, Alessia 1740 

Ciarleglio, Maria 743, 1731, 1899 

Cicalese, Luca 1991 

Ciccarese, Francesca 355 

Cillo, Umberto 148, 351, 356, 1270 

Cinar, Resat 1374 

Ciociaro, Demetrio 2209 

Cipriano, Madalena Z. 1921 

Cirincione, Brenda 720, 728 

Citti, Caitlin C. 380, 438, 457 

Civan, Jesse M. 10, 172, 1452, 1906 

Claassen, Mark 1007


Clahsen, Thomas 963 

Clair, Heather B. 666, 2233 

Clarett, Danisa M. 1257 

Clària, Joan 2087 

Claridge, Lee C. 2143 

Clark, Virginia C. 138, 839 

Clasen, Kristen M. 213 

Claudel, Thierry 977 

Clavien, Pierre A. 499, 665 

Cleary, Sean P. 350 

Clemens, Mark G. 1023 

Clément, Marc-André 51, 1520, 1523 

Cloherty, Gavin A. 1797 

Cloonan, Yona K. 1555 

Clouston, Andrew D. 784, 1362 

Coakley, Dion F. 1467, 1478, 1485 

Coakley, Eoin 36 

Cobanoglu, Murat C. 193 

Cobbold, Jeremy F. 2190 

Cocchio, Silvia 629 

Cocchis, Donatella 1 

Codrington, John F. 1846 

Coelho, Henrique Sergio M. 273, 289 

Coenraad, Minneke 2069 

Coffin, Carla S. 1576 

Cogliati, Bruno 586 

Cohen, Alice 1034 

Cohen, Ari J. 818, 1242 

Cohen, Ayelet 1235 

Cohen, Daniel E. 1039, 1088 

Cohen, David E. 322, 881, 900, 942 

Cohen, Eric 1039 

Cohen, Jose L. 215 

Cohen-Naftaly, Michal 875 

Cohn, Jennifer 1826 

Coilly, Audrey 3, 95, 1237, 1269, 

1782 

Cojuhari, Lilia 31 

Colan, Juan 2084 

Cole, Banumathi K. 664 

Colecchia, Antonio 741 

Coleman, Carla 1864, 1880 

Collado, Sol 175, 1916 

Colle, Isabelle 305 

Colledge, Danni 1557 

Collier, Jane 2190 

Collins, Allan J. 1867, 1883 

Collins, Christine 705 

Collins, Jenna 1050 

Collins, Peter 564 

Colmenero, Jordi 860 

Colnot, Nathalie 887 

Colombo, Bruno S. 2075 

Colombo, Massimo 229, 393, 1049, 

1086, 1743, 1747, 2011, 2012 

Colvin, Richard 2257, 2259, 2260, 

2261 

Combalia, Andres 615 

Comeglio, Paolo 882, 902 

Comerford, Megan 2166 

Compagnon, Philippe 215 

Compan, Anita 1154 

Concepcion, Waldo 1208 

Concepción, Mar 734, 1509 

Conde, Isabel 1230 

Conde de la Rosa, Laura 1382 

Cong, Min 187, 1404 

Congiu, Mario 996 

Congly, Stephen 1576 

Congly, Stephen E. 204 

Connelly, Margery A. 2227 

Conner, Elizabeth A. 677 

Conrads-Frank, Annette 1089 

Considine, Aisling B. 646, 1098, 1113, 

1744 

Conti, Filomena 30, 95, 997, 1239, 

1409 

Conti, Maria 309, 1153 

Contreras, Patricia C. 1315, 1522 

Conway, Brian 40, 249, 1161, 1198, 

1892 

Cook, Darrel 86 

Cooke, Graham 1826 

Coombes, Jason D. 163, 1978, 2143 

Cooper, Curtis 204, 249 

Cooper, David 1083 

Cooper, James N. 2173 

Cooper, Matthew A. 55 

Cooper, Stewart 1598 

Copeland, Neal 234 

Coppel, Ross L. 77, 613 

Coppleson, Yael J. 1454 

Coppola, Carmine 1802, 1844 

Coppola, Nicola 431, 1543, 1837 

Corbelli, Jody 435 

Corbett, Christopher 1321 

Corbin, Ian 361 

Cordero, Paula 1022, 1281 

Cordoba, Juan 1509 

Corey, Kathleen E. 154, 2166, 2194 

Cornberg, Markus 37, 252, 1058, 

1131, 1559, 1572, 1861, 2026 

Cornelissen, Jan 1803 

Cornella, Helena 253 

Cornella, Scott L. 1501 

Cornide-Petronio, Maria Eugenia 342 

Cornillie, Alexia 2079 

Corouge, Marion 1194 

Corpechot, Christophe 73, 634, 642 

Correia, Lurdes 473 

Correia, Pedro 473 

Correnti, Jason 1314 

Corrigan, Margaret 636 

Corsa, Amoreena C. 1678 

Corsello, Tiziana 1991 

Cortes, Andrea C. 2118 

Cortesi, Paolo 571, 1453 

Cortez-Pinto, Helena 150, 832, 897, 

940, 960 

Coseano, Hernan 1752 

Coskuner, Seher A. 1622 

Costa, Cilénia B. 150, 1876 

Costa, Fernando G. 2221 

Costa, José N. 473 

Costentin, Charlotte E. 132, 373, 417, 

421, 449, 1232 

Costes, Laurent 567 

Cotler, Scott 532, 559, 982, 1172, 

1248, 1854 

Cotoi, Corina G. 1733 

Cotrim, Helma P. 2237 

Cotte, Laurent 568, 1093 

Cotterell, Adrian 838 

Coughlan, Diarmuid 636 

Courcambeck, Jerome 394 

Cournand, Henri 1813 

Cousien, Anthony 1845 

Cousin, Vladimir 1728, 1735 

Couto, Claudia A. 2200 

Covington, Kyle 129 

Covinsky, Kenneth E. 841 

Cowie, Benjamin C. 1566, 2060 

Cox, Eleanor 1480, 1524 

Cox, I. Jane 1300 

Cox, Josiah 352 

Coyle, Catelyn 1810 

Coyne, Karin 1467, 1485 

Crabb, David W. 1332 

Craig, Kevin 2116 

Crawford, Darrell H. 818, 975 

Craxi, Antonio 1179, 1802, 1844 

Creighton, Chad 129 

Crespo, Javier 378, 1511, 2032, 2171 

Crespo Yanguas, Sara 586 

Crissien, Ana Maria 108 

Cronberg, Alexandra 113 

Crosbie, Orla M. 1103 

Crosby, Jeff R. 183, 1353 

Cross, Jennifer 2007 

Crothers, Michael W. 13 

Crouchet, Emilie 1019 

Cruz, Helder 1921 

Cruz, Pedro E. 1921 

Cruz, Ricardo 971 

Csak, Timea 512 

Csizmadia, Eva 1394 

Cubero, Francisco Javier 828, 1779, 

1924 

Cuervas-Mons, Valentin 1180 

Cuervo, Ana Maria 333 

Cui, Jeffrey Y. 45, 913, 1425, 1431 

Cui, Julia Yue 655 

Cui, Qingwen 1546, 1854 

Cui, Wei 85, 1031, 1785 

Cui, Yuanyuan 2107 

Culberson, Cathy 1389 

Cullaro, Giuseppe 272 

Culver, Emma L. 637 

Cummings, Oscar 160, 948 

Cummings, Yvette 1111, 1155, 1164 

Cunha, Aloisio S. 2200 

Cuperus, Frans 808 

Curry, Michael P. 704, 1045, 1046, 

1049, 1108, 1177 

Cursaro, Carmela 1888 

Curth, Harald M. 687 

Curti, Marco 337, 1388 

Curtis, Donna 1737 

Curtis, Timothy 71, 843 

Cusi, Kenneth 2147, 2208 

Cutilli, Carolyn 536 

Cutler, Murray J. 424 

Cuzin, Lise 568, 1093 

Cyr, Wendy 2116 

Czaja, Albert J. 303 

Czaja, Mark J. 1333 

Czul, Frank 1054, 2256 

D’Albuquerque, Luiz C. 469 

D’Aliberti, Deborah 1127 

d’Alteroche, Louis 3, 95, 145, 206 

D’Amico, Ronald 1067 

D’Souza, Rashmi 1733 

da Silva, Rita de Cássia M. 469 

da Silva, Tereza Cristina 586 

Da Silva, Hannah 2238, 2254 

da Veiga, Zulane 762, 2072, 2076 

Dadabhai, Alia S. 1749 

Daelken, Nicole 200 

Daffis, Stephane 35, 2009 

Dahari, Harel 982, 1854


Daher, Saleh N. 1878 

Dahlan, Yaser 1607 

Dai, Chia-Yen 398, 2264 

Dai, Erhei 209, 1587 

Dai, Feng 16 

Dai, Shenglong 1583 

Dai, Wing-chiu 70, 1746 

Daiber, Andreas 950 

Daikoku, Atsuko 517 

Daikos, Georgios L. 2094, 2095 

Daily, Michael F. 854 

Daita, Kalyani 905, 1309, 1527, 2249 

Daix, Valérie 974 

Dal Ben, Matteo 2098 

Dale, Cheryl 647 

Dalekos, George N. 73, 76, 2012 

Daley, Donnele 1282 

Dalgard, Olav 40 

Dalgic, Buket 1719 

Dalla Valle, Fabio 767 

Dalton, Harry 523 

Daltro, Carla 2237 

Daly, Ann K. 225, 1930, 1933 

Dametto, Ennia 1 

Damjanov, Nevena 521 

Dammermann, Werner 1629 

Dams, Els 1846 

Dan, Yunjie 1865 

Dandekar, Satya 63 

Danelski, Lisa 1839 

Danford, Christopher J. 2166 

Dannhorn, Emily 489, 490 

Danso, Hassan 987 

Dantas-Correa, Esther B. 2075 

Dao, Doan Y. 1555 

Dao, Thong 1072, 1187, 2082 

Dar, Sadaf 1005 

Dara, Lily 2135 

Darling, Jama M. 217, 1057 

Daruich, Jorge 438, 2106 

Das, Kausik 750 

Das, Kshaunish 750 

Das, Sukanta 581, 1344 

Das, Suvarthi 962, 2133, 2137 

Dasarathy, Jaividhya 2162, 2186 

Dasarathy, Srinivasan 330, 334, 518, 

651, 667, 668, 671, 1308, 1468, 

2162, 2182, 2186 

Dasgupta, Aditi 1825 

Dasgupta, Kathleen 1229 

Dash, Ajit 175, 664, 915, 1916 

Dash, Asha 1096 

Dash, Srikanta 501, 506, 1096 

Dash, Suchismita 1911 

DaSilvera, Eduardo 1549 

Dattaroy, Diptadip 962, 2133, 2137 

Daud, Amna 71, 563, 843, 847, 849 

Daugherty, Tami 1182, 1189 

Daulouede, Jean-Pierre 1796 

Davalos-Moscol, Milagros B. 595 

Davenport, Mark 134, 1691 

David, Lawrence 2188 

David, Paul 1663, 2088 

David, Waseem 358 

Davidson, Brian 1373 

Davidson, Nicholas O. 911, 1361, 

1365 

Davies, Nathan 1300 

Davies, Sean S. 1407 

Davies, Susan 258 

Davila, Jessica A. 357 

Davila, Marta L. 130, 682 

Davis, Brian C. 213, 1768 

Davis, Eric 1178 

Davis, Roger J. 1924 

Davuluri, Gangarao 330, 334, 518, 

651, 667, 668, 671, 1308 

Dawson, George 1797 

Dawson, Paul 19 

Day, Christopher P. 2164, 2183 

Day, Gretchen 626 

Day, James W. 776 

Daye, John 1716 

Dayoub, Rania 954 

de Andrade, Mariza 607, 610 

de Araujo Horcel, Lucas 57 

de Assuncao, Thiago 692, 1355, 1406 

de Avila, Leyla 2205 

de Boer, Bastiaan 2185 

de Boer, Ian H. 2228 

de Branco, Gabriel 1484, 1494, 1495 

De Carlis, Luciano 1751 

de Carvalho, Juliana R. 273, 289 

De Chiara, Francesco 908, 1517 

De Coppi, Paolo 337, 1388 

De Francesco, Raffaele 2011 

De Gaetano, Anna Maria 432 

de Galocsy, Chantal 1091 

De Gottardi, Andrea 2087 

De Gramont, Armand 417 

de Jong, Koert P. 1434 

De Jong, Ype P. 1841 

de Knegt, Robert J. 445, 492, 1007, 

1166, 1196, 1585, 1586, 1590, 

1998, 2002, 2003 

De La Cruz-Munoz, Nestor 914 

De la Flor-Robledo, María 1758 

de la Grange, Pierre 1288 

de la Motte, Carol A. 2132 

de la Peña, Joaquin 745 

de la Revilla, Juan 1511, 2012 

de la Rosa, Brauley 1870 

De La Rosa, Guy 1040 

de la Vega-Prieto, Maria Jose 1342 

de Ledinghen, Victor 3, 95, 117, 206, 

251, 450, 777, 1439, 1796, 1847, 

2181 

De Luca, Francesca 222 

de Magnée, Catherine 1734 

de Man, Robert A. 68, 1590, 1803 

De Man, Joris G. 976 

De Maria, Vincenzo 1802, 1844 

De Martin, Eleonora 487, 1782 

de Mendoza, Carmen 1180, 1881, 

1908 

de Mesquita, Fernanda C. 326, 1418 

De Minicis, Samuele 21 

De Nicola, Stella 1747 

de Niet, Annikki 1995, 2046 

de Oliveira, Jarbas R. 1418 

De Pasquale, Paola 1493 

de Stefano, Giorgio 431 

De Vito, Claudio 221 

de Vos, Marie 1216 

de Vos, Paul 1427 

de Vree, J. Marleen 208, 2263 

de Vries, Annemarie C. 68 

de Vries, Elisabeth M. 74, 604, 623 

de Vries, Marianne 1238 

de Vries, Niek 637 

de Waart, Dirk R. 604 

De Winter, Benedicte Y. 894, 976 

De Wolf, Andre M. 9 

De-Oertel, Shampa 1080, 1105, 1120 

Deal, Barbara J. 401 

Deans, Julie 1290 

Debes, Jose 572, 1752 

Debette-Gratien, Marie Line 95 

Debray, Dominique 1728 

Debzi, Nabil 1621 

Decaens, Thomas 421, 449 

Decaris, Martin 1425 

Deckmyn, Olivier 786 

Deemer, James 527 

Defreitas, Andrew 401 

Degallaix, Nathalie 974 

Degrassi, Cristina 2098 

Degré, Delphine 1091, 2079 

Deguchi, Yoshio 1941 

DeHart, David N. 319 

Deheragoda, Maesha 1711, 1712, 

1721 

Dehovitz, Jack 1444 

Deichsel, Danilo 245 

Dejong, Cornelis H. 818 

Dekhtyar, Tanya 705 

Del Bello, David P. 1202 

Del Rio Hernandez, Armando E. 337, 

1388 

dela Cruz, Rouchelle D. 2214 

Dela Cruz, Anna Christina 914, 1267, 

2081 

Delaney, William E. 35, 1594, 2009 

Delasalle, Patrick 1072 

Delaunay, Jean-Louis 196 

Delelo, Rolland 1409 

Delemos, Andrew 17, 2166 

Deleuran, Bent 301 

Delgado, Evan 493 

Delio, Maria 943 

Delitto, Anthony 1244 

Dell’Olio, Dominic 1 

Della Corte, Cristina 435 

Deltenre, Pierre 1818 

Delvart, Valérie 1237 

Delwaide, Jean 1091 

DeMasi, Ralph 1163 

Demetris, Anthony J. 216, 1736 

DeMino, Mary 1650 

Demir, Kadir 478 

Demir, Münevver 2195 

Demiroz, Ali Pekcan 2051 

Demma, Shirin 1904 

DeMorrow, Sharon 454, 621, 835, 

1507 

Demzik, Alysen 1235 

den Dulk, A. Claire 1238 

Denarie, Michel F. 1085 

Deneau, Mark 1702 

Deng, Yanghong 1731 

Deng, Yanhong 743, 1899 

Deng, Yong-Qiong 1564, 1602 

Denis, Jacques 2082 

Denning, Jill M. 746, 1049, 1128 

Deny, Paul 1162 

DePaoli, Alex M. 106, 198, 627, 1986 

DePeralta, Danielle K. 957 

Derbala, Moutaz F. 1147, 1188 

Derderian, S. C. 98 

Desai, Archita P. 1459


Desai, Payal 1801 

Desai, Sachin 2261 

DeSanto, Jennifer 1849 

Deshmukh, Sandeep 1906 

Desmond, Paul V. 442, 996 

Desnick, Robert J. 2111, 2112 

Deterding, Katja 1131, 2003 

Detlefsen, Sönke 780, 789, 1440 

Deuffic-Burban, Sylvie 1845 

Deutsch, Melanie 2019 

Deutz, Nicolaas E. 671, 1523 

Dev, Anouk T. 442, 483 

Devara, Anupama 420 

Devarajan, Karthik 371 

Devarbhavi, Harshad 284, 2101 

Dever, John 549 

Devereaux, Michael W. 1687, 1688, 

2134 

Devière, Jacques 178 

Devoto, Marcella 1693 

Devue, Cécile 887, 2148 

Devun, Daniel 1242 

DeWitt, Sheila H. 143 

Dhali, Gopal K. 750 

Dhaliwal, Sandeep S. 639 

Dhamija, Ravinder Mohan 147 

Dhanasekaran, Renumathy 129, 1965 

Dharancy, Sebastien 95, 263, 1269, 

1808, 2222 

Dhawan, Anil 134, 1691, 1733 

Dhersin, Jean-Stéphane 1845 

Dhillon, Amar P. 793, 1530 

Dhillon, Ammen P. 2007, 2034 

Dhillon, Sonu 774 

Dhiman, Radha K. 214 

Di Benedetto, Antonino 1476 

Di Benedetto, Fabrizio 1751 

Di Bisceglie, Adrian M. 94, 121, 1057, 

1478, 1598 

Di Carlo, Antonio 1122 

Di Cecco, Sara R. 836 

Di Cocco, Silvia 958 

Di Gregorio, Vincenza 2067 

Di Leo, Alfredo 1802, 1844 

Di Maio, Velia Chiara 222 

Di Maira, Giovanni 973 

Di Maira, Tommaso 1230, 1251 

Di Marco, Mariella 355 

Di Marco, Vito 973 

Di Martino, Vincent 3, 95, 145, 206, 

554, 1036, 1187, 2082 

Di Pace, Maria 595 

Di Paolo, Daniela 44 

Di Paolo, Daniele 203, 222 

Di Spirito, Mike 730 

Diago, Moisés 2032, 2171 

Diago, Teresa 1271 

Diaz, Giacomo 166, 1646 

Diaz, Maria Alba 860, 1889 

Diaz-Meco, Maria T. 232 

Dickerman, Richard 1169 

Dickmeyer, Laura J. 401 

Dickson, Rolland C. 580, 1329, 1831, 

2073 

Dickstein, Aaron 1592 

Dieguez, Gabriela 1069 

Diehl, Anna Mae 66, 80, 161, 737, 

889, 936, 962, 1386, 1435, 1715, 

2137, 2143, 2169, 2188 

Diener, Katrina 503 

Dieterich, Douglas 199, 380, 457, 545, 

726, 1046, 1065, 1079, 1090, 1095, 

1106, 1108, 1112, 1151, 1185, 

1202, 1538 

Dietz, Julia 980, 1559 

Difrancesco, Robin 332 

Dige, Anders 301 

Dik, Ebru 1622 

Dillon, Jayne 11 

Dillon, John F. 225, 1930 

Dimova, Rositsa B. 1823, 1841 

Dincses, Elif 2180 

Ding, Dora 616, 624 

Ding, Jia 366, 935 

Ding, Jiguang 2048 

Ding, Qian 1355 

Ding, Shitao 1865 

Ding, Wen-Xing 140, 598, 1325 

Ding, Xiwei 1966 

Ding, Yanhua 2058 

Ding, Yifeng 140 

Dinh, Phillip 241, 2021, 2025 

Dippold, Christine 197 

Disabato, Mara 215 

Dispenzieri, Angela 1263 

Divine, George 1254 

Diwanji, Rohan 510, 2116 

Do, Ailinh 2250 

Do, Albert 283 

Doan, Sylvia 1702 

Dodge, Jennifer L. 4, 50, 53, 462, 

1273, 1444 

Doehle, Brian 91, 205, 219, 1168 

Dohmen, Kazufumi 2031 

Doi, Hiroyoshi 1302, 1941 

Doi, Yoshinori 1201 

Doignon, Isabelle 816 

Dole, Kiran 2259, 2260 

Dolganiuc, Angela 839, 1286 

Dolgin, Natasha 845 

Dolin, Christine E. 587 

Dollé, Laurent 2143 

Dombrowicz, David 928 

Domenicali, Marco 1493 

Dominguez, Edward A. 1169 

Dominitz, Jason A. 128 

Donato, Francesco 1568 

Donato, Maria F. 1743, 1747, 1751 

Donchev, Vladamir 1479 

Donde, Hridgandh 1331 

Dondossola, Daniele E. 1743, 1751 

Donepudi, Ajay C. 885 

Dong, Chao 1526 

Dong, Tien S. 271 

Dong, Weiyan B. 1832 

Dongiovanni, Paola 973, 2187 

Dongre, Neelesh 260, 753 

Donner, Aaron J. 507 

Donohue, Julie M. 104, 151 

Donohue, Terrence M. 589, 1389 

Donovan, John 1350 

Donovan, Julie 36 

Doo, Edward 213, 218, 2150, 2220 

Doorenspleet, Marieke E. 637 

Dopazo, Hernán 2217 

Dore, Gregory 40, 91, 543, 1083, 

1161, 1868 

Doria, Cataldo 1276 

Dorko, Kenneth 1291 

Doshi, Dilesh 1189 

Doss, Wahid H. 708, 1076, 1163 

Dossier, Claire 196 

Dostal, David E. 830 

Dottin, Stephania 1452 

Dou, Xiaoguang 2048 

Dougherty, Michelle 1864, 1880 

Dourakis, Spyros P. 1489, 2096 

Douschan, Philipp 2068 

Doussau, Adelaide 132 

Dove, Lorna M. 531, 550 

Doyle, Adam 11 

Doyle, Erin H. 229, 988, 1289, 1742 

Dranoff, Jonathan A. 1384, 1387 

Dreher, Bernd 1170, 1200 

Drenth, Joost 105, 162, 293, 609, 

2102, 2145 

Drescher, Hannah K. 963 

Dreymueller, Daniela 100 

Drinane, Mary 1406 

Drobnik, Ann 1823 

Du, Huijuan 638 

Du, Sandie Y. 2007 

Du, Xiaofei 2061 

Duan, Na-Na 935 

Duan, Weijia 374 

Duan, Xiaoqiong 981 

Duan, Yuyou 685 

Duan, Zhong-Ping 209 

Duarte, António 897 

Duarte, Ignacio 2210 

Duarte, Sebastião M. 2221 

Duarte-Rojo, Andres 445, 1099, 1166, 

1307, 1471, 1481, 1857 

Duarte-Salles, Talita 170 

Dubay, Derek 1250 

Dubin, Sterling M. 529 

Duboc, Denis 1194 

Dubray, Clarisse 394 

Dubreuil, Marta 615 

DuBrock, Hilary M. 9 

Duca, Ana Maria 1180 

Ducancelle, Alexandra 1439 

Duchesne, Léa 1833 

Duclos-Vallee, Jean-Charles 3, 95, 1237, 

1782 

Ducom, Julie 529, 549 

Dudek, Hank 510, 2116 

Dueñas, Eva 378 

Dufour, Jean-Francois 221, 425, 445, 

1049, 1166, 1884, 2164 

Dugas, Lara R. 1248 

Dugum, Alex 855, 856 

Dugum, Mohannad 855, 856 

Duksal, Faysal 2219 

Dumitru, Raluca 1327 

Dumont, Laure 98 

Dumortier, Jérôme 3, 95, 1158, 2082 

Duncan, Andrew W. 493 

Duncan, Oliver J. 848 

Duncan, Richard G. 36 

Dundas, Pauline 1104 

Dunn, Michael A. 648, 1244, 1468, 

1754 

Dunn, Winston 1814, 1815 

Dunnington, Katherine M. 725 

Dupont, Benoît 179 

Duran, Domingos 1859 

Duran-Struuck, Raimon 2129 

Durand, Christine M. 1749


Durand, Francois 95, 265, 480, 772, 

887, 1288, 1781, 2148 

Durand-Schneider, Anne-Marie 196 

Duranyildiz, Derya 478 

Durchschein, Franziska 2068 

Durel, Ryan M. 1242 

Durkalski, Valerie L. 213, 1766 

Durkin, Marian E. 677 

Duron, Cédric 1505 

Duseja, Ajay K. 214 

Dusheiko, Geoffrey M. 712, 2059 

Dutta, Amal K. 815 

Dutta, Chaitali 510, 2116 

Dutta, Sandeep 710, 719, 723 

Dutton, N. 2233 

Duvivier, Claudine C. 568, 1093 

Duvoor, Chitharanjan 1307 

Duvoux, Christophe 3, 95, 421, 449, 

1232 

Dvorak, Karel 1487 

Dvory, Hadas S. 247 

Dvory-Sobol, Hadas 91, 219, 718, 

1168 

Dweik, Nazeeh Z. 1147, 1188 

Dybala, Claire 2175 

Dyson, Jessica K. 611 

Eason, James 1212, 1233 

Eaton, John E. 607 

Ebinuma, Hirotoshi 181, 569, 2131, 

2140, 2234 

Ebisutani, Yusuke 906 

Eccleston, Jason L. 1498 

Eckert, Christoph 1299, 1385 

Eckert, Doerthe 1066 

Eckmann, Lars 1319 

Eda, Keisuke 1724 

Edelman, Devorah 943 

Edelman, Elazer R. 1292 

Eder, Michael 297, 1070 

Edlin, Brian R. 1042 

Edula, Raja G. Reddy 295, 1181 

Edwards, Angela 1063 

Edwards, Jeffrey 1915 

Edwards, Michael 503 

Efe, Cumali 308 

Egawa, Mayumi 227, 1974 

Egert, E. M. 521 

Egetemeyr, Daniel P. 1183 

Eghtesad, Bijan 840, 850, 855, 856, 

1265, 1271 

Eguchi, Akiko 57, 112, 1371, 1522, 

1971 

Eguchi, Junichi 1302, 1941 

Eguchi, Yuichiro 1097, 1632, 1819, 

2202, 2207 

Ehman, Richard 1355 

Ehrinpreis, Murray N. 420 

Ehrlich, Adam C. 765 

Ehrlich, Laurent 830 

Ehsan, Nermine 1862, 1879, 1890 

Einar, Bjornsson S. 1926 

Eisner, Sigal 875 

Ekbom, Anders 116 

Ekong, Udeme D. 1731 

Eksteen, Bertus 76, 616, 2143 

El Aggan, Hoda 1011 

El Deeb, Nevine M. 1011 

El Demiry, Sally 1011 

El Kasmi, Karim C. 1687, 1688, 1692, 

2134 

El Raziky, Maissa 1163 

El Refaie, Ahmed 1890 

El Sabaawy, Maha 1890 

El-Bendary, Mahmoud 1236 

El-Fert, Ashraf Y. 733 

El-Gazzaz, Galal 850, 1271 

El-Matary, Wael 1702 

El-Saadany, Mohamed M. 758 

El-Serag, Hashem B. 90, 156, 244, 

357, 1048 

El-Sherif, Omar 1103 

Elam, April D. 1835 

Elashry, Amr 1853 

Elbadri, Mohammed E. 1147 

Elbanna, Abduh M. 2215 

Elbasha, Elamin 727 

Eley, Timothy 720, 728 

Elfeki, Mohamed 1236 

Elfeky, Sarah 534 

Elgilani, Faysal 2 

Elia, Chiara 266 

Elizaga, Jaime 1503 

Elkhammas, Elmahdi 874, 1260, 1280 

Elkhashab, Magdy 1051, 2004, 2025 

Elkrief, Laure 145, 264, 772 

Elliott, Michael 106, 627 

Elmazaly, Mohamed A. 1862, 1879 

Elorza, Ainara 930 

Elsabaawy, Maha M. 1862, 1879 

Elsayad, Elham 1147 

Elshamy, Ahmed M. 229 

Elsharkawy, Aisha 1163 

Elsheikh, Elzafir 316, 892, 1186 

Elzohry, Hasan A. 733, 1853 

Eminler, Ahmet T. 846 

Emmert-Buck, Michael R. 166 

Emond, Jean C. 369, 838, 1454 

Emson, Claire 1425 

Enders, Felicity B. 605 

Endig, Jessica 2128 

Endo, Mizuki 2267 

Endo, Ryujin 1528 

Endo, Takeshi 135 

Enejosa, Jeffrey 1065, 1084 

Enestvedt, C. Kristian 69, 481 

Eng, Francis J. 229 

Eng, Lauren M. 2214 

Eng, Rene S. 2214 

Engelen, Marielle 671 

Engelmann, Cornelius 261, 276 

Engelmann, Michael 1002 

Engle, Ronald E. 166, 1646 

English, Shirley 1104 

Enomoto, Hirayuki 466, 770, 1533, 

1669, 1975 

Enomoto, Masaru 429, 1125, 1824 

Enomoto, Nobuyuki 406, 1027, 1565, 

1673 

Enooku, Kenichiro 461, 912 

Enrich, Carlos 64 

Enriquez, Aimee 1478 

Ensor, Katherine 1991 

Enweluzo, Chijioke U. 748, 769 

Epstein, Sara 414 

Erario, Madeline 575 

Erdelyi, Katalin 1374 

Erensoy, Selda 1739 

Ergen, Can 1756 

Erhart, Wiebke 617 

Erne, Elke M. 1802, 1844 

Ersoy, Baran A. 942 

Ersoz, Galip 246, 1247, 1739 

Erwin, Patricia J. 282 

Escajadillo, Natali 734 

Escalante, Shannon M. 278 

Escheik, Carey 534 

Escobedo, Miguel 1281 

Eshelman, Anne 556 

Eskind, Lon 17 

Eslam, Mohammed 2185 

Eslick, Guy D. 1343, 1450 

Esmadi, Mohammad 748, 769 

Esmat, Gamal E. 708, 1076 

Español-Suñer, Regina 98 

Espera, Hannah G. 1798, 1807 

Espinosa-Cuevas, Angeles 1471 

Espiritu, Christine 33, 2064 

Esplugues, Juan V. 1913 

Esposito, Isabella 1180 

Esquivel, Carlos O. 341, 1208, 2123 

Esteban, Rafael 1021, 1674, 2012 

Estep, James M. 316 

Estep, Michael J. 956, 1186 

Estes, Chris R. 1859 

Estévez, Pamela 2171 

Estrabaud, Emilie 998, 1601 

Eswaran, Sheila L. 1215 

Etschmaier, Alexandra 1466 

Ettorre, Giuseppe M. 1751 

Etzion, Ohad 779, 1498, 1720, 2144, 

2224 

Eubanks, Haleigh B. 1384 

Eun, Hyuk-Soo 1358, 1414 

Eun, Jong Ryeol 685 

Evans, Alison 371 

Evans, Helen M. 1728 

Evelyn, Zoehrer 1726 

Everson, Gregory T. 630, 746, 1045, 

1049, 1065, 1130, 1849 

Evon, Donna M. 1063, 1114 

Ewing, Micheal 193, 197 

Ezaz, Ghideon 283 

Fabbrizzi, Alessio 1190 

Faber, Klaas Nico 591, 612, 1398, 

1416 

Fabre, Thomas 1380 

Fabrellas, Nuria 266 

Fabri, Milotka J. 2004 

Fabris, Luca 76, 571, 811, 1950 

Facchetti, Floriana 435, 2011, 2012 

Facciuto, Marcelo 870 

Factor, Valentina M. 677 

Fadel, William 1903 

Fadin, Mariangela 351 

Fagan, Kevin 784 

Fagiuoli, Stefano 571 

Fagoaga, Omar 1254 

Faisal, Nabiha 204 

Faivre, Sandrine 417 

Falck-Ytter, Yngve 1154 

Falcon-Perez, Juan M. 953 

Faletti, Riccardo 2209 

Falguières, Thomas 196 

Falissard, Bruno 373 

Falkner, Keith C. 666, 1303, 2142, 

2233 

Fallon, Michael B. 79, 82, 570, 743, 

1424, 1470, 1513, 2151 

Fallowfield, Jonathan A. 260, 753, 

2168


Faloppi, Luca 435 

Falzano, Loredana 1802, 1844 

Famoso, Giulia 268 

Fan, Daiming 468, 754 

Fan, Daping 2133 

Fan, Jia 366 

Fan, Yang-Yi 56 

Fang, Yun 2204 

Fang, Zhengfeng 194, 1689 

Fanning, Liam J. 1103 

Fante, Garrett 1111, 1164 

Fantini, Damiano 1988 

Farag, Raghda 1236 

Farah, Taha 1607 

Farci, Patrizia 166, 1646 

Farella, Nunzia 431 

Farges, Olivier 373 

Fargion, Silvia 393, 2187 

Farhang Zangneh, Hooman 350 

Farhat, Nada 2262 

Farhood, Anwar 586 

Farinati, Fabio 351, 355, 356 

Färkkilä, Martti A. 76, 633 

Farnan, Jeanne M. 544 

Farooqi, Javed I. 1872 

Farrell, Geoffrey C. 55, 230, 1909 

Farrokhi, Kaveh 1012, 1297 

Farrugia, Helen 442 

Farrugia, Maria 2251 

Farsad, Khashayar 481 

Fartoux, Laetitia 417 

Fasano, Massimo 2003 

Fasola, Carlos 1169 

Fasolato, Silvano 148, 266, 292 

Fasseu, Magali 1288 

Fateen, Waleed 791, 800 

Fatela, Narcisa 150 

Fattovich, Giovanna 1166, 1568, 1802, 

1844 

Fauler, Günter 1726 

Fausther, Michel 1384 

Fawcett, Jonathan 818 

Fazel, Yousef 2192, 2212 

Feaver, Ryan E. 915 

Fedchuk, Larysa 206 

Federman, Alex D. 1792 

Fehr, Jan 1818 

Fei, Ran 1850, 1877, 1973 

Feigh, Michael 978 

Feilen, Nicole A. 1389 

Feld, Jordan J. 348, 350, 445, 542, 

714, 1051, 1067, 1086, 1099, 1107, 

1166, 1556, 1797, 1998 

Feldbrügge, Linda 1222, 1394, 2227 

Felder, Martina M. 355 

Feldman, Amy G. 137, 1714, 1738 

Feldman, Brian M. 1713 

Feldner, Ana Cristina A. 1484, 1494, 

1495 

Feldstein, Ariel E. 55, 57, 112, 969, 

1371, 1522, 1971, 2185 

Feldstein, Vickie A. 1570 

Felga, Guilherme 469 

Felix, Sean C. 2204 

Félix, Jorge 1859 

Felizarta, Franco 41, 1065 

Feng, Bo 1436, 1850, 1877 

Feng, Chun 731 

Feng, Dan 1323, 1347 

Feng, Dechun 182, 2179 

Feng, Hwa-Ping 725, 730, 731 

Feng, Sandy 841, 852, 1736 

Feng, Wenke 114, 1305, 1306 

Feng, Ziding 371 

Feng, Zongdi 1703 

Fenkel, Jonathan M. 1452 

Fenlon, Laura 1455 

Feranchak, Andrew P. 815 

Feray, Cyrille 987, 1123 

Ferenci, Peter 37, 252, 297, 714, 732, 

1058, 1067, 1070, 1092, 1137, 1885 

Ferguson, Beatrice 2154 

Ferguson, James W. 631, 1216, 1234, 

1710 

Ferlitsch, Arnulf 149, 270, 475, 732, 

755, 771, 1466 

Ferlitsch, Monika 1466 

Fernandes, Flavia F. 84, 762, 2072, 

2076 

Fernandes, Prabhavathi 2246 

Fernandez, Alejandro 2084 

Fernandez, Javier 266, 2084 

Fernández, David 953, 2244 

Fernández, Natalia 1511 

Fernandez Mena, Carolina 1295, 1520 

Fernandez-Bermejo, Miguel 2171 

Fernández-Carrillo, Carlos 1511 

Fernandez-Checa, Jose 899, 1320 

Fernandez-Hernando, Ana 65 

Fernandez-Hernando, Carlos 65 

Fernandez-Montero, Jose V. 1881, 1908 

Fernández-Moreira, Daniel 896, 939 

Fernandez-Rodriguez, Camino Maria 

1341 

Fernández-Rodríguez, Conrado M. 2171 

Fernández-Varo, Guillermo 659 

Fernandez-Zapico, Martin E. 692 

Fernando, Samantha 527 

Ferolla, Silvia M. 2200 

Ferrante, Shannon A. 727 

Ferrarese, Alberto 268, 351, 1270, 

1889 

Ferrari, Carlo 1802, 1844, 2015, 2052 

Ferrari, Maria de Lourdes A. 2200 

Ferrari, Teresa C. 2200 

Ferrari-Lacraz, Sylvie 1735 

Ferraris, Pauline 501, 506, 1096 

Ferraz, Maria Lucia 48, 1484, 1494, 

1495 

Ferreira, Carlos R. 1722 

Ferreira, Paula 150 

Ferrell, Linda 1736 

Ferrer, Maria Teresa 378 

Ferri, Nicola 2187 

Fessel, W. Jeffrey 2015, 2052 

Festi, Davide 741 

Fevery, Bart 1040 

Fevery, Johan 76 

Fey, Holger 1294, 1370, 2124 

Feyssa, Eyob L. 530, 535 

Fialla, Annette D. 780, 789, 1440 

Fibbi, Meghan F. 1136 

Fickert, Peter 662, 2068 

Fiel, M. Isabel 29, 253, 254, 870, 988, 

1228, 1289, 1364, 1397, 1742, 2124 

Fierer, Daniel S. 29, 89, 1090, 1094, 

1112, 1852 

Fierro, Nora A 1855 

Figler, Robert 175, 1916 

Figorilli, Francesco 309, 1153, 1300, 

1769, 1780, 1781 

Figueiredo, Fatima F. 84, 2189 

Figueiredo, Sabrina R. 2237 

Figueruela, Blanca 2032 

Filippi, Sandra 882, 902 

Filomia, Roberto 288, 1127, 1476 

Fimmel, Claus J. 1791 

Fink, Michael A. 442, 483 

Finnemann, Claudia 1287 

Fiore, Francesco 222 

Fiorotto, Romina 811 

Firpi, Roberto J. 839 

Fischer, Hans-Peter 2115 

Fischer, Janett 1638 

Fischer, Josiane 2075 

Fischer, Sandra E. 11, 2184, 2238, 

2254 

Fischl, Margaret 1444 

Fisher, Jonathan S. 861, 872 

Fisher, Robert A. 71, 843, 847 

Fisher-Hoch, Susan 1424 

Fishman, Sarah 1875 

Fitzgerald, Katherine A. 180 

Fitzhugh, Courtney 1498 

Fitzmorris, Paul S. 1220, 1253 

Fitzpatrick, Emer 1711, 1733 

Fitzpatrick, Thomas 577 

Fizanne, Lionel 2188 

Flaherty, John F. 243, 1678, 2004, 

2059 

Flake, Sebastian 412 

Flamm, Steven L. 702, 739, 1045, 

1046, 1049, 1108, 1130, 1442 

Flanigan, Colleen 573 

Flavell, Richard A. 1284 

Fleckenstein, Jaquelyn 2091 

Fleming, Michael F. 1231 

Fletcher, Linda 784 

Fletcher, Simon P. 35, 2009 

Flisiak, Robert 241, 243, 1502, 2014 

Flood, Michael C. 790 

Floreani, Annarosa 73, 76, 299, 609, 

629, 634 

Flores, Lalo 33, 2064 

Flores Molina, Manuel 1380 

Flores-Toro, Joseph A. 177 

Florman, Sander S. 253, 870, 988, 

1228, 1289, 1397 

Fluker, Shelly-Ann 1835 

Fogacas, Homero S. 762 

Fognani, Elisa 1190 

Fok, Brandon 462 

Foldes, Emily 275 

Folino, F. 268 

Folkert, Ian W. 868 

Fombuena, Blanca 1863 

Fondevila, Constantino 860 

Font-Burgada, Joan 1971 

Fontaine, Hélène 206, 1194, 1808 

Fontana, Robert J. 213, 225, 706, 

1768, 1850, 1877, 1922, 1923, 

1930, 1931, 1932, 1933 

Fontanges, Thierry 1072 

Foote, Patrick H. 169 

Forbes, Kevin F. 531 

Forbes, Stuart J. 62, 678 

Ford, David A. 65 

Ford, Jo-Ann E. 647 

Ford, Joel S. 153


Ford, Mary 1801 

Forde, Kimberly A. 122, 536, 865, 

871, 1470 

Forne, Montserrat 378 

Forns, Xavier 700, 703, 746, 982, 

1045, 1049, 1084, 1086, 1130, 1889 

Forsythe, Jordan M. 1804 

Fortea, Jose Ignacio 1520 

Fosby, Bjarte 819 

Foschi, Francesco G. 355, 435 

Foskett, Pierre 1721 

Foss, Aksel 819 

Foster, Andrew L. 89, 1090 

Foster, Graham R. 167, 241, 249, 

1098, 1173, 1179, 1571, 1626, 2014 

Foti, Michelangelo 665 

Fouchard-Hubert, Isabelle 117, 206 

Foucher, Juliette 450, 1796, 2181 

Fougerou-Leurent, Claire 3 

Fourati, Slim 999 

Fournier, Claire 1997 

Fowler, Kathryn 2150 

Fox, Caroline S. 2191 

Fox, Rena K. 16 

Fracanzani, Anna Ludovica 393, 2187 

Fraeman, Kathy 1175 

Frampton, Gabriel A. 1507 

Franceschet, Irene 73, 299, 629 

Francescucci, Angelica 2160 

Franchitto, Antonio 621, 817, 835 

Francis, Fadi 1753 

Francis, Heather L. 190, 621, 812, 817, 

822, 835, 929, 1317, 1349, 1363 

Francis, Prashanth 1210 

Francis, Susan 1480, 1524 

Francisco Ziller, Nicki M. 836 

Francisco-Recuero, Irene 1028 

Franco, Edson S. 526, 1784 

Franco, Ricardo A. 1804 

Francoz, Claire 265, 772, 1781 

Francque, Sven M. 105, 162, 797, 

894, 928, 976, 1091, 2145 

Frank, Brombacher 183, 1353 

Frank, Wacker 444 

Franke, Olivia 626 

Frankel, Angela 1680 

Frankova, Sona 787 

Franques, François 450 

Fransen, Floris 1427 

Fransen, Paul 976 

Frantsen, Marijke 527 

Franzè, Maria Stella 288 

Fraser, Andrew 1104 

Frassineti, Giovanni Luca 435 

Frater, Yvonne 424 

Frazier, Lynn M. 94, 1057 

Frederic, Oberti 117 

Frederick, R Todd 538, 1467, 1485, 

2104 

Fredericks, Emily M. 133 

Fredrick, Linda 722 

Freedman, Daniel 36 

Freiberg, Matthew 1488 

Freise, Chris 838 

Freissmuth, Clarissa 297, 732, 1070, 

1092, 1137, 1885 

Freitas, Luiz Antônio R. 2237 

Frelin, Lars 992 

French, Audrey 1444 

French, Benjamin 853 

French, Dorothy 632, 1359, 1367, 

2149 

French, Pim 1637 

Frenette, Catherine T. 4, 7, 108, 861, 

872 

Frenguelli, Luca 337, 1388 

Frey, Christian 2009 

Freyer, Nora 1921 

Fried, Michael 46 

Fried, Michael W. 94, 217, 1034, 

1057, 1114, 1598, 1826 

Friedman, Harley 1329, 2073 

Friedman, Nir 672 

Friedman, Scott L. 29, 131, 1095, 

1185, 1337, 1364, 1392, 1405, 

1779, 2174 

Frigo, Anna Chiara 148 

Frissen, Mick 100 

Frode, Tania S. 2075 

Fröhlich-Reiterer, Elke 1726 

Frossard, Jean-Louis 2174 

Fryzek, Nancy 218 

Fu, Bo 1051 

Fu, Dong 1909 

Fu, Rongdang 353, 375 

Fu, Sherry 1850, 1877 

Fu, Yan-Ling 1616 

Fu, Yiming 1536 

Fu, Yu 328 

Fuchs, Bryan C. 957, 1938 

Fuchs, Michael 52, 1041, 1472 

Fuentes, Javier 378, 2032 

Fujie, Satomi 513, 1912, 1987 

Fujii, Hideki 1956, 2125, 2156, 2207 

Fujii, Hironobu 906, 2156 

Fujii, Yohei 1059 

Fujikawa, Kazuyuki 1679 

Fujiki, Masato 1271 

Fujimori, Shunji 291 

Fujimoto, Go 707 

Fujimoto, Jiro 188, 466, 513, 781, 

1420, 1987 

Fujimoto, Toyoshi 880 

Fujinaga, Hidetaka 912 

Fujino, Hatsue 986, 1056, 1649, 1681, 

1944 

Fujisaka, Yasuyuki 1026, 1648, 1943, 

2167 

Fujisawa, Koichi 329 

Fujisawa, Toshio 505 

Fujitani, Naoki 1375 

Fujiwara, Kei 1954 

Fujiwara, Naoto 461 

Fujiya, Mikihiro 927 

Fujiyama, Kazuhito 1642 

Fujiyama, Shunichiro 1191, 2020 

Fukazawa, Kyota 878 

Fukubayashi, Kotaro 1987 

Fukuhara, Takayuki 472, 1056, 1893 

Fukui, Hiroyuki 1201, 2213 

Fukuizumi, Kunitaka 304, 1159 

Fukumitsu, Ken 1976 

Fukumura, Yuki 820 

Fukunishi, Shinya 966 

Fukushima, Hirofumi 1771 

Fukushima, Kentaro 869 

Fukutomi, Keisuke 1584 

Funaki, Masaya 495, 994, 1000 

Fung, James 70, 1545, 1550, 1746, 

2000 

Fung, Phoenix 295 

Fung, Scott 241, 1678, 2004, 2014, 

2025, 2184, 2238, 2254 

Furlage, Rosemary 332 

Furlini, Giuliano 1888 

Fürst, Dieter O. 80 

Furuichi, Yoshihiro 1047 

Furuta, Kunimaro 227, 453, 1974 

Furuya, Shinji 1956, 2125 

Fusai, Giuseppe 1407 

Fusco, Dahlene 991, 1015, 1630, 

1644 

Fushimi, Kazumi 392 

Fuster, Josep 860 

G. Lavoie, Elise 1387 

Gaarde, William A. 2135 

Gabr, Reham Mohamed 1821 

Gacusan Munson, Marie Lou 1920, 

1929 

Gaeta, Giovanni B. 241, 1802, 1844, 

2014 

Gaetano, John N. 553 

Gaggar, Anuj 243, 1551, 1557, 1563, 

1594, 1651, 1668, 2014, 2015, 

2037, 2043, 2052, 2059 

Gaggini, Melania 2209 

Gaglio, Paul J. 1141 

Gagnieu, Marie-Claude 1158, 1162 

Gahl, William A. 1722 

Gaidosh, Gabriel S. 914 

Gailer, Ruth E. 1455 

Gaisa, Michael 89 

Galanis, Kostas 2012 

Galati, Joseph S. 1057 

Galbraith, James W. 1804 

Gale, Michael 28 

Galen, Kelly 1266 

Galil, Karin 36 

Gallay, Philippe 2035 

Galle, Peter R. 376, 1949 

Gallego, Adolfo 378 

Gallego-Durán, Rocío 2171 

Galli, Andrea 949, 1963 

Galli, Claudio 1888 

Galli, Silvia 1888 

Galloway, Christopher 2247 

Galmozzi, Enrico 2011 

Gama-Carvalho, Margarida 960 

Gambato, Martina 982, 1889 

Gambino, Carmine G. 288 

Gambino, Roberto 2209 

Gamil, Mohamed 1163 

Gamil, Mostafa 1076 

Gan-Schreier, Hongying 1411 

Gandhi, Yash 720, 728 

Gane, Edward J. 38, 40, 219, 243, 

249, 713, 746, 1045, 1049, 1083, 

1120, 1128, 1130, 1557, 1563, 

1651, 1678, 2004, 2015, 2052, 

2059, 2225 

Ganesan, Murali 589 

Ganesh, Shanthi 2116 

Ganger, Daniel 211, 401, 1759, 1761 

Ganguli, Anirban 871 

Ganjoo, Naveen 2170 

Ganne, Nathalie 1847 

Ganne-Carrié, Nathalie 421, 449, 2082 

Ganz, Michael 1456, 1461 

Gao, Bin 110, 115, 182, 334, 903, 

1580, 2179


Gao, Bing 1080, 1105, 1120 

Gao, Guangping 496 

Gao, Rui 698 

Gao, Xiuzhu 1149, 1603, 1657, 1869 

Gao, Yanhang 182, 1580, 1620 

Gao, Zhiliang 120, 1423, 1772, 2048 

Gaouar, Farid 642 

Garabedian, Elizabeth 1717 

Garas, George 848, 2099 

Garassini, Miguel E. 595 

Garcia, Gabriel 1182, 1189 

Garcia, José H. 469 

Garcia, Maria 1251 

Garcia, Mina A. 1874 

Garcia, Ruel 1549 

García, Elisabet 2071 

Garcia-Alonso, Veronica 2087 

Garcia-Buitrago, Monica 914 

Garcia-Caldero, Hector 1518 

García-Cortés, Miren 595 

Garcia-Martinez, Irma 979, 1284, 2179 

García-Monzón, Carmelo 930, 2171 

Garcia-Muñoz, B. 595 

Garcia-Pagan, Juan Carlos 326, 745, 

1518 

Garcia-Ruiz, Carmen 899, 1320 

García-Ruiz, Inmaculada 896, 939 

Garcia-Saenz de Sicilia, Mauricio 1307 

García-Samaniego, Javier 1028, 2171 

García-Torres, María Luisa 2171 

Garcia-Tsao, Guadalupe 82, 570, 743, 

745 

García-Valdecasas, Juan Carlos 860 

Gardenier, Donald 1151, 1185, 1870 

Garimella, Tushar 720, 728 

Garioud, Armand 554, 2082 

Garner, Jessica S. 929, 1317, 1349 

Garnier, Muriel 417 

Garre, Carmen 378 

Gartner, Michael 725, 730, 731 

Garvin, Jennifer H. 529 

Gasbarrini, Antonio 355, 432, 1802, 

1844 

Gasmi, Imène 693 

Gaspar, Rui 1540 

Gassler, Nikolaus 100, 828, 1924 

Gastaldelli, Amalia 2209 

Gatselis, Nikolaos 2012 

Gatta, Angelo 148 

Gauci, James Mario 2251 

Gaudio, Eugenio 621, 817, 834, 835 

Gault, Nathalie 145 

Gautam, Dheeraj 2245 

Gautam, Manjushree 1146, 1199 

Gautherot, Julien 196 

Gauthier, Aline 1176, 1205 

Gautsch, Sebastian 80 

Gavasso, Sabrina 351 

Gavis, Edith A. 52, 1463, 1491, 1527 

Gawrieh, Samer 436, 1903, 2166 

Gayed, Yasser 1275 

Gazzin, Silvia 2098 

Ge, Dongliang 1651, 1668, 2037 

Ge, Fengxia 972 

Ge, Jingjing 1396 

Ge, Xiadong 1382, 2124, 1940 

Gedaly, Roberto 673, 854, 1267, 2081 

Geerts, Anja M. 1091 

Geevarghese, Sunil K. 1223 

Geffers, Robert 1659, 2128 

Gehrke, Nadine 1949 

Geier, Andreas 37, 46, 2195 

Geiser, Mary 78 

Geldenhuys, Werner J. 1338 

Gelli, Maximiliano 487 

Gelman, Ekaterina 1597 

Gelson, William 258 

Gely, Cristina 1509 

Genco, Petrina V. 1236 

Genda, Takuya 1851 

Genescà, Joan 742, 1509 

Genovese, Domenico 1153 

Genovese, Federica 1437 

George, Jacob 119, 441, 1077, 1868, 

2185 

Gerada, Jurgen 2251 

Gerada, Martina 2251 

Geraghty, Estella M. 630 

Geratikornsupuk, Nopavut 360 

Gerber, Naomi Lynn 316 

Gerken, Guido 1779 

German, Polina 1133, 1707 

Germani, Giacomo 1270, 1889 

Gershwin, M. Eric 77, 602, 606, 613, 

619, 621, 639, 640, 1285 

Gerstoft, Jan 251 

Geskus, Ronald 74, 623 

Gevers, Tom J. 293 

Geyer, Peter R. 1170, 1200 

Ghabril, Marwan 1467, 1478, 1903, 

1931, 1932 

Ghali, Peter 204 

Ghalib, Reem H. 715, 739, 1040, 

2239 

Ghanekar, Anand 11 

Ghanta, Mythili 1122 

Ghany, Marc G. 218, 1598, 1722 

Ghare, Smita 1331 

Gharib, Ahmed M. 1498 

Gharibian, Derenik 1135 

Ghesquiere, Wayne 1161 

Ghobrial, R. Mark 1215 

Ghosh, Dipu 269 

Ghosh, Martha S. 521 

Ghosh, Siddhartha 1521 

Ghoshal, Kalpana 99, 493 

Ghoz, Hassan M. 169, 397 

Gi, Young Jin 130, 682 

Giabicani, Mikhael 1288 

Giama, Nasra H. 127, 169, 1965 

Gianella, Sara 1852 

Giannakeas, Nikos 801 

Giannelli, Valerio 265, 2067 

Giannini, Edoardo G. 355 

Giannone, Ferdinando A. 1493 

Gianserra, Laura 222 

Giard, Jeanne-Marie 1259 

Gibert, Cynthia 1488 

Gibson, Neil 208, 2263 

Gijbels, Marion J. 922 

Gil, Ana Isabel 1028 

Gil, Gregorio 804 

Gilbert, Melissa 1693 

Gill, Kirat 1111 

Gill, Ryan M. 157, 236, 2161 

Gill, Upkar S. 167, 1571, 1626 

Gillberg, Per-Göran 810 

Gilles, HoChong 52 

Gillespie, Brenda W. 838 

Gillevet, Patrick M. 905, 1463, 1527 

Gillies, Robert D. 29 

Gilmartin, Jocelyn 730 

Gilmour, Kimberly 1723 

Gilroy, Richard 565, 837, 1814, 1815 

Gines, Pere 266, 860, 1327, 2069, 

2087 

Giordani, Maria Teresa 1803 

Giostra, Emiliano 2174 

Giovanni, Shewit 1229 

Girala, Marcos A. 595 

Girard, Pierre-Marie 1676, 1677, 1833 

Girardin, Francois 1539 

Girgrah, Nigel 1242 

Girish, Veeranna 284 

Gish, Robert 32, 237, 1111, 1155, 

1164, 1549, 2022 

Giugliano, Silvia 1430, 1706, 1708 

Giuily, Nathalie 2044, 2045 

Giuliante, Felice 432 

Giusto, Michela 2067 

Gjuka, Donjeta 2151 

Glaser, Shannon S. 190, 621, 817, 

830, 834, 835, 929, 1317, 1349, 

1978 

Glässner, Andreas 1287 

Glebe, Dieter 2026 

Gloria, Helena 1256 

Glorioso, Jaime M. 2 

Go, Kristina L. 177 

Gobejishvili, Leila 1312, 1334 

Goblet, David 731 

Godlewski, Grzegorz 1374 

Goel, Amit 1475, 1817 

Goel, Aparna 545, 1337 

Goel, Satyender 563, 849 

Goeser, Felix 1287 

Goeser, Tobias 687 

Goetz, Matthew B. 1488 

Goetze, Oliver 46 

Goetzmann, Jason E. 32, 2022 

Gogela, Neliswa A. 1667 

Gogoi, Naminita 1344 

Gogos, Charalambos 2094, 2095 

Goh, Boon-Bee George 451 

Goh, Evan 47 

Gokturk, Huseyin Savas 2206 

Gola, Anna 1455 

Gola, Elisabetta 148, 292 

Golabi, Pegah 153, 1186, 2173, 2192 

Goldberg, David J. 1794 

Goldberg, David S. 5, 9, 427, 563, 

837, 849, 853, 864, 865, 868, 1218, 

1470, 2083 

Goldberg-Clein, Tova 1878 

Golden, Aaron 515 

Golden-Mason, Lucy 28, 503, 938, 

1430, 1706, 1708 

Goldenberg, Anna 1713 

Goldkamp, William J. 1212 

Goldstein, David B. 1651, 2037 

Goldstein, Eric 556 

Golin, Carol E. 1114 

Gomaa, Asmaa 344, 1163, 1853 

Gomberoff, Leon 1796 

Gomel, Rachel 630 

Gomes, Marilia B. 2189 

Gomes-Gouvêa, Michele S. 48 

Gómez Rodríguez, Rafael 2032 

Gomez-Camarero, Judith 2171 

Gomez-Moreno, E.M. 583


Gonçalves, Afonso 150 

Gonçalves, Nilza 1876 

Gong, Guo-Zhong 2023 

Gong, Jiao 120 

Gong, Li 1526 

Gong, Yuewen 361 

Gonzales, Gabriel R. 1146, 1199 

Gonzalez, Adam 774 

Gonzalez, Adriano m. 1484, 1494, 

1495 

Gonzalez, Frank J. 110 

Gonzalez, Patricia 982 

Gonzalez, Stevan A. 1146, 1199 

González, Javier M. 745 

González Ballerga, Esteban 2106 

Gonzalez-Aldaco, Karina 1855 

González-Jiménez, Andrés 595, 596 

Gonzalez-Reimers, Emilio 1341, 1342 

Gonzalez-Rodriguez, Agueda 930 

Good, Steven S. 2266 

Goodman, Zachary D. 624, 632, 737, 

739, 891, 892, 956, 1359, 1428, 

1435, 2149, 2173, 2192 

Goodrich, Nathan P. 1694 

Goossens, Nicolas 364, 1539, 1742, 

2174 

Gooz, Monika 319 

Gordon, Fiona H. 564 

Gordon, Fredric D. 412, 1106 

Gordon, Ronald E. 1364 

Gordon, Stuart C. 15, 106, 249, 525, 

627, 722, 1079, 1128, 1538, 1578, 

1789, 1800, 2015, 2052 

Goree, Jessica R. 1384, 1387 

Gores, Gregory J. 127, 381, 415, 825, 

895, 1124, 1315, 1994 

Gorgievski, Meri 1575 

Goria, Odile 1187 

Gorn, Chad 1452 

Gorsh, Boris 1456, 1461 

Gosselin, Nathalie H. 1915 

Gossmann, Mona 272 

Gotlieb, Neta 1685 

Goto, Fumio 164, 406 

Goto, Hidemi 411, 443, 880, 967, 

1061, 1167 

Goto, Kaku 505 

Goto, Takaaki 1573, 1599, 1618 

Gottfried, Michelle 211, 1759, 1766, 

1777 

Gottfriedova, Halima 787 

Gotthardt, Daniel 76, 618 

Gottwald, Mildred D. 2247 

Gou, Eric 2100, 2111, 2112 

Goukos, Dimitrios 2094, 2095 

Gould, John 1969, 2139 

Goulis, Ioannis 1126, 2012 

Goumard, Claire 1239, 1409 

Gounder, Prabhu P. 125, 1790, 1798, 

1807 

Gourari, Samir 1621 

Gournay, Jerôme 2082 

Goutte, Nathalie 373 

Gouw, Annette S. 1728 

Gove, James E. 1790, 1798, 1807 

Govil, Sanjay 867 

Gow, Paul 442, 483, 842, 1077, 1225 

Goyal, Vik 532, 559 

Gozgit, Joseph M. 1966 

Grabmeier-Pfistershammer, Katharina 

561, 1204 

Gracia-Sancho, Jordi 8, 326, 342, 

1418, 1518 

Graf, Rolf 499, 665 

Graffner, Hans 810 

Gragnani, Laura 1190 

Graham, Camilla S. 1826 

Graham, Emily 648 

Gralla, Jane 2085 

Grammatikopoulos, Tassos 134, 1691, 

1721 

Granato, Celso 48 

Grande, Lourdes 2171 

Grange, Jean Didier 1158 

Grangé, Jean-Didier 2082 

Grant, Charlotte R. 300, 608 

Grant, Claire 1480 

Grant, David 11, 53, 838 

Grant, Stephanie 1507 

Grant, Tiffannia 785 

Granzow, Michaela 80 

Grasset, Denis 264 

Graupera, Isabel 266 

Graw, Frederik 982 

Graziadei, Ivo 1092, 1885, 2087 

Grbec, Matjaz 292, 767 

Greaves, Wayne 727 

Grebely, Jason 40, 1083, 1868 

Greco, Stephanie 1282 

Green, Kile J. 611 

Green, Pamela 128 

Green, Richard M. 658 

Greenblatt, Ruth 1444, 2252 

Greene, Laurence 1141 

Greene, Tom 2112 

Greenhalgh, Stephen 62 

Greenstein, Andrew E. 1367 

Greenup, Astrid-Jane 1243 

Greenwood, Jeremy R. 957, 1938 

Gregori, Josep 1021, 1674 

Greig, Paul D. 11, 53 

Greinert, Robin A. 1519 

Greisen, Stinne 301 

Grenier, Carole 889 

Gress, Jacqueline 210 

Grewal, Priya 1095, 1185 

Grewal, Thomas 64 

Grieco, Antonio 432 

Grieco, Stefania 983 

Griesemer, Adam D. 2129 

Griffin, Sean 1243 

Griffiths, Jenifer 521 

Grigorieva, Julia 1900 

Grimm, Andrew A. 98 

Grimm, Dirk 98 

Grint, Paul 208, 2263 

Gripshover, Janet 1101 

Grochowski, Christopher 1693 

Groen, Albert 604 

Groessl, Erik J. 529, 549 

Grogan, Martha 1263 

Gronbaek, Henning 119, 301, 2071, 

2087 

Groothuis, Geny M. 1427 

Grossmann, Maria 1594 

Grossmann, Mathis 842 

Grove, Jane 225, 908 

Grønbæk, Henning 768 

Grundhoff, Adam 1629 

Grünhage, Frank 1429 

Gruss, Hans J. 2169 

Gschwantler, Michael 37, 252, 1058, 

1179, 1885, 1886 

Gu, Daehoe 1438 

Gu, Jiezhun 597, 1922, 1926, 1931 

Gu, Yurong 1772 

Gualano, Gisela 595 

Gualdieri, Luciano 1543 

Guan, Bo-Jhih 668 

Guañabens, Nuria 615 

Guardascione, Maria Anna 742 

Guarner, Carlos 734 

Guarner-Argente, Carlos 2084 

Guarneri, Valeria 1888 

Guarrera, James V. 71, 843, 847 

Guasti, Daniele 902 

Gubertini, Guido A. 222 

Guerin, Coralie L. 887 

Guerra, Bernadette 32, 2022 

Guerra, Giselle 1148 

Guerra, Juan Francisco 1217 

Guerra, Vito 437 

Guerrier, Micheleine 190 

Guerrieri, Francesca 165 

Guessab, Nawal 1621 

Guest, Rachel V. 678 

Guevara, Asdrubal 1281 

Gufraind, Alexander 1854 

Guglielmo, Flavius 1276, 1906 

Guha, Indra Neil 624, 1524, 2168 

Guicciardi, Maria Eugenia 825 

Guillot, Adrien 693 

Guimarães, Raquel a. 84 

Guixé-Muntet, Sergi 326, 1418 

Gulamhusein, Aliya 600, 610 

Gulati, Rishabh 1827 

Guldiken, Nurdan 831 

Gundogdu, Ceren 932 

Gunneson, Tim 293 

Gunsar, Fulya 246, 308, 1247, 1739 

Guo, Grace L. 919, 934 

Guo, Haifeng 1867, 1883 

Guo, Jinsheng 131, 1405 

Guo, Junli 1977 

Guo, Luyang 1947 

Guo, Qing 1611, 2002 

Guo, Simin 2002 

Guo, Wei 1660 

Guo, Wengang 754 

Guo, Xiao-Lin 1465 

Guo, Xin 1651 

Guo, Zifang 725 

Gupta, Ekta 1005, 1268 

Gupta, Nidhi 223 

Gupta, Nitika A. 12 

Gupta, Parul 1664 

Gupta, Priya 1917 

Gupta, Sanjeev 340, 515, 1764, 1917 

Gupta, Soumi 1189 

Gupta, Subhash 853 

Gupta, Tarana 214 

Gupta, Tripti 1242 

Gurakar, Ahmet 1214, 1255, 1749 

Gurel, Selim 2059 

Gustot, Thierry 178, 1091, 2079 

Guthke, Reinhard 954 

Guthrie, Deanne 1804 

Guthrie, Gregory J. 194, 1689 

Gutic, Enisa 1886


Gutierrez, Julio A. 39, 247, 1433, 

1900 

Gutkind, J. Silvio 20 

Guy, Cynthia D. 157, 161, 2169, 2188 

Guy, Jennifer 4, 462 

Guyader, Dominique 206, 1808, 1847 

Guyer, William 1041 

Guzman, Carlos A. 1762 

Guzman, Grace 1266, 1940 

Guzzardi, Maria Angela 672 

Gwak, Geum-Youn 422, 423, 485, 

1591, 1957, 1960, 2049 

Gydesen, Sofie 2178 

Gyongyosi, Benedek 496, 1318, 1330 

Ha, Yeonjung 446, 448 

Haagmans, Bart L. 2122 

Haber, Barbara A. 42, 133, 701 

Habersetzer, François 1158 

Habib, Adil 1169 

Habib, Aida 1391 

Habib, Nagy 118 

Habib, Naomi 672 

Habib, Robert 118 

Habic (Suciu), Alina 741 

Habra, Reema 556 

Hadengue, Alexandra 417 

Hadi, Salah 208, 2263 

Hadj-Nacer, Khaled 554 

Hadzic, Nedim 1711, 1712, 1723, 

2105 

Hadziyannis, Emilia 2019 

Hafler, David 1731 

Haga, Hiroaki 335, 358 

Haga, Yuki 1207, 1628 

Hagan, Michael 528 

Hagège, Hervé 2082 

Hagen, Susan J. 322 

Hagey, Lee R. 23, 829 

Hagihara, Atsushi 1125, 1824 

Hagiwara, Hideki 1201 

Hagström, Hannes 158, 308 

Hahn, Judith A. 1793 

Hahn, Katherine 286 

Hai, Hoang 191, 1125, 1824 

Hai, Seikan 466, 1420 

Haider, Samran 420 

Haifer, Craig 1225 

Haigh, W. G. 230, 923 

Hainaut, Pierre 170 

Hairwadzi, Henry N. 1667 

Haiying, Sun 2261 

Hajarizadeh, Behzad 1868 

Hajelssedig, Omer E. 1147 

Hajifathalian, Kaveh 1265, 1894 

Hakobyan, Syune 1891 

Halazun, Karim J. 369 

Halbower, Ann C. 1715 

Hale, Pamela 193, 197 

Halegoua-De Marzio, Dina 1261, 1276, 

1452 

Halff, Glenn A. 1900 

Halfon, Philippe 394, 1009 

Halilbasic, Emina 76 

Hall, Andrew R. 337, 793, 1388, 1530 

Hall, Coleen 708 

Hall, Russell 2169 

Hallal, Hacibe 583 

Hallberg, Par 225 

Haller, Wolfram 1728 

Halliday, John 622 

Hallinan, Erin K. 2220 

Hallsworth, Kate 2183 

Halm, Ethan 548 

Halseth, Amy 2175 

Halsted, Charles H. 2109 

Hamano, Mina 796 

Hamdaoui, Nabila 693 

Hamdeh, Shadi 748, 769 

Hameed, Bilal 160, 236, 1777 

Hametner, Stephanie 1092, 1885 

Hamid, Saeed 1872 

Hamilton, James P. 1749 

Hamilton, Thomas 668 

Hammad, Radi 1076, 1163 

Hammami, Muhammad B. 1535 

Hammond, Rachel 1551, 1563 

Hampe, Jochen 617 

Han, Chang 185, 686 

Han, Guo Rong 209 

Han, Guohong 468, 754, 851 

Han, Ho Seong 857, 1902 

Han, Huanzi 1370 

Han, Hui 588 

Han, Jude D. 1366 

Han, Juqiang 1595 

Han, Kwang-Hyub 91, 346, 504, 1080, 

1105, 1168, 1542, 1558, 1581, 1605 

Han, Lingling 249 

Han, Ma Ai Thanda 1720, 2144 

Han, Meifang 1656, 1683, 1684 

Han, Ping 328 

Han, Seungbong 446 

Han, Sheng-Na 1284, 2179 

Han, Steven-Huy B. 488, 2015, 2052 

Han, Yuan-Ping 904 

Han, Yuyan 190, 454, 835, 929, 

1317, 1349 

Han, Zhou 709 

Hanczko, Robert 1961 

Hand, Fiona M. 1216 

Handa, Priya 946 

Hanes, Justin 115 

Hanf, Rémy 105, 162, 974, 2145 

Hangartner, Thomas N. 1694 

Hanje, A. James 213, 874, 1260, 

1280, 1759 

Hann, Hie-Won 371, 1554, 2004 

Hannan, Nicholas R. 258 

Hanouneh, Ibrahim A. 840, 855, 856, 

1215, 1265 

Hansen, Bettina E. 68, 73, 76, 348, 

445, 601, 609, 634, 1166, 1196, 

1568, 1585, 1586, 1590, 1998, 

2001, 2002, 2003, 2012, 2013 

Hansen, Janne F. 780, 789, 792, 1440 

Hansi, Navjyot K. 167, 1571 

Hanson, Jeffrey 166 

Hanson, John 722 

Hao, Ke 253, 254 

Hao, Ying 451 

Hara, Tasuku 886, 2230 

Hara, Yuichi 995, 1937, 2199 

Harada, Masaru 593, 883, 1819 

Haraguchi, Masafumi 1671 

Harbers, Marten 208, 2263 

Harbonnier, Jean 1796 

Hardesty, Josiah 666 

Hardie, Claire 611 

Hardison, Regina M. 1714 

Hardtke, Svenja 1572 

Hardwick, James P. 925 

Hardy, Timothy 1408 

Hargrove, Laura 812, 822, 1363 

Haridy, James 311 

Harif, Yael 875 

Hariharan, Siddharth 316 

Hariri, Susan 1787 

Harms, Maren H. 73, 634 

Harmsen, Martin C. 1416 

Harmsen, William S. 397 

Harney, Sarah 136 

Harnois, Denise M. 50, 53, 1236, 1245 

Harper, Ann M. 861, 872 

Harrell, Sherrie M. 1099, 1857 

Harrigan, Richard 1832 

Harriman, Geraldine 957, 1938 

Harrington, Andrew 253 

Harris, Aaron M. 1574 

Harris, Edward N. 507 

Harris, Melissa 726 

Harrison, Stephen A. 105, 162, 737, 

1428, 1435, 2145, 2151, 2239 

Harrison-Findik, Duygu Dee 60 

Harry, Kathy M. 829 

Hartl, Astrid 2068 

Hartl, Johannes 523 

Hartleif, Steffen 1728 

Hartman, George D. 33, 2064 

Hartman, Joshua 1095 

Hartman-Neumann, Sandra 709 

Hartmann, Bolette 1689 

Hartmann, Dagmar 1170, 1200 

Hartmann, Phillipp 59 

Harty, Alyson 1095, 1151, 1185, 1202 

Haruna, Yoshimichi 173 

Harvey, Brandon K. 323 

Harvey, Ray 1050 

Harwood, H. James 957, 1938 

Hasan, Mohsin 954 

Hasan, Raza 1906 

Hasanaliyeva, Nigar 161, 2169 

Hasanin, Mohsen 1249, 1250 

Hasebe, Takumu 927 

Hasegawa, Daisuke 1392, 1405 

Hasegawa, Kunihiro 466, 770, 781, 

1533, 1669 

Hashem, Mohamed 1142 

Hashiba, Tomomi 699, 1953 

Hashimoto, Etsuko 1326, 2207 

Hashimoto, Koji 1271 

Hashimoto, Satoru 408, 783, 1600 

Hashimoto, Shigeatsu 551 

Hashmi, Haroon 36 

Haslett, Patrick 36 

Hasnain, Nadeem 1268 

Hassan, Ammar 1791 

Hassan, Mohamed A. 1101, 1184 

Hassanein, Tarek I. 41, 106, 627, 714, 

1120, 1813 

Hassany, Mohamad 708, 1076, 1163 

Hassona, Ehab M. 1011 

Hatano, Etsuro 402, 1213, 1918, 1976 

Hatooka, Masahiro 472, 1893 

Hatting, Maximilian 828 

Hattori, Noburiro 1446 

Hatzoglou, Maria 668 

Hau, Hans-Michael 1222 

Hauber, Joachim 1629 

Haufe, William 45, 913 

Hausding, Michael 950


Hawley, Richard C. 521 

Hay, Christopher 753 

Hayakawa, Masako 918 

Hayashi, Eiichi 1302, 1941 

Hayashi, Kazuhiko 411, 443, 880, 

967, 1061, 1167 

Hayashi, Norio 1197, 1201 

Hayashi, Paul H. 14, 533, 1922, 1923, 

1926, 1931, 2093 

Hayashi, Sanae 1655 

Hayashi, Takehiro 699, 1000, 1953 

Hayashi, Tomoyuki 699, 1953 

Hayata, Yuki 382 

Haydar, Ghada 1909 

Haydel, Brandy M. 988, 1289 

Hayden, Hubert 1525 

Hayes, C. Nelson 986, 1627, 1645, 

1649, 1681, 1858, 1895, 1944, 2120 

Hayes, Don 1280 

Hayes, Nelson 663, 1150, 1670, 1679 

Hayes, Peter C. 260, 753 

Haynes, Meagan 670, 1979 

Hazari, Sidhartha 1096 

Hazra, Saswati 239 

He, Aiwu R. 2118 

He, Chuangye 754 

He, Dengming 1865 

He, Hua 245 

He, Jia 919 

He, Lina 1945 

He, Xiao-Song 77, 602, 613, 639 

He, Xiuting 1580, 1620 

He, Yingli 1541, 1579, 1617, 1773 

He, Yong 182 

Heathcote, E. Jenny 348 

Heaton, Nigel 212, 1733 

Heba, Elhamy 45, 2150, 2201 

Heckmann, Melanie 1726 

Hede, Shalini 1456, 1461 

Hedskog, Charlotte 219 

Heegsma, Janette 612, 1398 

Hegde, Pushpa 1391 

Hegge, Julia O. 32, 2022 

Heidrich, Benjamin 1002, 1572 

Heier, Eva-Carina 1299, 1385 

Height, Susan 1711, 1723 

Heikenwälder, Mathias 2128 

Heim, Markus H. 221, 1884 

Heimbach, Julie 6, 50, 53, 836, 837, 

1226, 1263 

Heinisch, Birgit B. 149 

Heinz, Stefan 331 

Hellard, Margaret 1083 

Heller, Theo 218, 779, 1498, 1650, 

1717, 1720, 2144 

Hellerstein, Marc 1425 

Hellmich, Brigitte 1886 

Helmke, Steve M. 1849 

Helmy, Housam 733 

Heluwaert, Frédéric 1072 

Hemmingsson, Tomas 158 

Henao, Ricardo 161 

Henderson, Mark 323 

Hendrikx, Tim 922 

Heneghan, Michael A. 212, 300, 312, 

608, 1098, 1744, 1758 

Hénique, Carole 887 

Henkel, Anne 257 

Henriksen, Kim 2178 

Henriques Abreu, Pedro 473 

Henry, Linda 18, 315, 316, 1035, 

1460, 2212 

Henry, Mitchel 1280 

Henry, Zachary 13, 761 

Hentati, Hassen 215 

Heo, Jeong 749, 876, 2028, 2038 

Heo, Nae-Yun 88, 1208, 2155 

Hepner, Ashley 1111, 1155, 1164 

Herath, Chaturika 1246 

Herber, Adam 276 

Heredia, Nancy 1770 

Herlong, H. Franklin 682, 2118 

Herman, Mark A. 2227 

Hernaez, Ruben 1214 

Hernandez, Brenda Y. 1553 

Hernandez, Carolyn 45, 913, 1425 

Hernandez, Céline 30 

Hernandez, Dennis 709 

Hernandez, Maria D. 457 

Hernández, Nelia 225, 595 

Hernandez-Alejandro, Roberto 837 

Hernandez-Guedea, Marco A. 1281 

Hernandez-Luis, Ruben 1341 

Hernandez-Rocha, Cristian 2210 

Hernandez-Santos, Nydiaris 1650 

Herrin, Ann 549 

Herrine, Steven K. 1452 

Hertel, Paula M. 133, 1694 

Herve, Camille 3 

Herzer, Kerstin 37, 252, 1058 

Heubi, James E. 1694 

Heuman, Douglas M. 52, 1463, 1472, 

1491, 1527 

Heurgué-Berlot, Alexandra 264, 308, 

1072 

Hew, Huong 1832 

Hewitt, Annette 1790, 1798, 1807 

Heymann, Felix 100, 1756 

Hezode, Christophe 42, 206, 714, 

1086, 1102, 1107, 1123 

Hézode, Christophe 1442 

Hiasa, Yoichi 338, 788, 1293, 2146 

Hibi, Taizo 336 

Hickey, Raymond D. 2 

Hickman, Matthew 1794 

Hickson, Ford 1830 

Hidaka, Hisashi 756 

Hidalgo, Juan 1295 

Hidvegi, Tunda 193, 197 

Hige, Shuhei 1116, 2065 

Higuchi, Kazuhide 966 

Higuchi, Nobito 1159 

Higuera, Monica 253 

Hijdra, Rosanne M. 89, 1112 

Hikita, Hayato 26, 97, 231, 453, 669, 

1082, 1197, 1357, 1635, 1636, 

1642, 1661, 1809, 1982, 1985, 

2130, 2136 

Hilburn, David 658 

Hildt, Eberhard 1559 

Hill, Andrew M. 1826 

Hillaire, Sophie 145 

Hillman, Luke 211 

Hilscher, Moira 605 

Hinkle, Greg 36 

Hino, Keisuke 995, 1632, 1937, 2199 

Hinrichsen, Holger 1078, 1435 

Hinson, Jack A. 1914 

Hinton, Alice 874 

Hiraga, Nobuhiko 663, 986, 1150, 

1627, 1645, 1649, 1670, 1681, 

1858, 1895, 1944, 2120 

Hirahara, Kazuki 1486 

Hiramatsu, Akira 472, 1893, 2199 

Hiramatsu, Katsushi 944, 1682 

Hiramatsu, Naoki 26, 231, 453, 669, 

1082, 1195, 1197, 1201, 1357, 

1635, 1636, 1809, 1982, 1985, 

2130, 2136, 2213 

Hiramatu, Akira 1056 

Hirano, Tadamichi 1420 

Hiraoka, Atsushi 524 

Hirashima, Noboru 1097 

Hirayama, Tasuku 995 

Hiriart, Jean-Baptiste 450, 1796, 2181 

Hirooka, Masashi 788, 1293 

Hirooka, Yoshiki 411, 880, 967, 1061, 

1167 

Hirose, Shunji 314, 383 

Hirota, Seiichi 781 

Hirsch, Amy 1154 

Hirsch, Geri 204, 647 

Hirschbein, Jonah 1479 

Hirschfield, Gideon 73, 76, 348, 607, 

631, 634, 636, 1234, 1457, 1710 

Hirsova, Petra 825, 895, 1315 

Hisatake, Garrett M. 538 

Hiththetiya, Kanishka 80 

Hittatiya, Kanishka 961, 2115 

Hjuler, Sara T. 2178 

Ho, Chanda 538 

Ho, Hsiu-Jon 345 

Ho, Ja-an Annie 654 

Ho, Samuel B. 59, 529, 549, 1041 

Hoang, Joseph K. 207, 359, 368, 386, 

388, 458, 1561, 1805, 1806, 1816, 

1848, 2250 

Hoang, Tanya 677 

Hobbs, Ursula 1452 

Hocking, Kyle M. 1223 

Hodge, Alexander 47, 945 

Hodson, James 574, 1728 

Hoefs, John C. 269 

Hoeke, Martijn O. 612 

Hoekstra, Mark 612 

Hoener zu Siederdissen, Christoph 

1131, 1803 

Hoerauf, Achim 2077 

Hoermann, Rudolf 842 

Hoevelmeyer, Nadine 1949 

Hofer, Harald 297, 732, 1070, 1092, 

1137, 1885 

Hoffman, Hal M. 1371 

Hoffmann, Udo 2191 

Hoffmann, Vera 687 

Hofker, Marten H. 922 

Hofmann, Alan F. 1915 

Hofmann, Wolf P. 1166 

Hogan, Brian J. 793, 1530 

Hohenester, Simon 609 

Höijer, Jonas 116 

Hoki, Toshifumi 1984 

Holder, Beth S. 300 

Hollabaugh, Kimberly 1094 

Holland-Fischer, Peter 1482 

Hollenbach, Marcus 1519, 1520 

Hollingsworth, Kieren G. 2183 

Holm, Kristian 76, 833


Holmberg, Scott D. 15, 525, 1789, 

1799, 1800 

Holmes, Benjamin 510, 2116 

Holmes, Jacinta A. 996 

Holmstom, Fredrik 992 

Holst, Jens 1689 

Holt, Andrew 1216, 1321 

Holt, Edward W. 306, 1252, 2159 

Holtzman, Deborah 525 

Holzmayer, Vera 1797 

Homan, Chriss E. 1790, 1798, 1807 

Homs, Maria 1021, 1674 

Honda, Akira 970 

Honda, Hiroki 310 

Honda, Koichi 2207, 2267 

Honda, Masao 144, 168, 325, 495, 

657, 699, 947, 990, 994, 1000, 

1010, 1014, 1348, 1377, 1395, 

1632, 1653, 1953, 1968, 1970 

Honda, Takashi 411, 443, 880, 967, 

1061, 1167 

Honda, Takuya 1671 

Hong, Jian 991, 1015, 1630, 1644 

Hong, Leena K. 1121 

Hong, Qiuting 606 

Hong, Sung Woo 1419 

Hong, Thai 442, 483 

Hong, Youngmi 876, 2028 

Honma, Yuichi 593, 883 

Hoofnagle, Jay H. 121, 218, 236, 597, 

779, 1598, 1922, 1923, 1926, 1931 

Hooker, Catherine A. 913, 2150 

Hooker, Jonathan 45, 913 

Hooper, Lora V. 111 

Hooshmand-Rad, Roya 317, 609, 625, 

628, 644 

Hoque, Rafaz 979, 2179 

Horban, Andrzej 2004 

Horberg, Michael A. 1811, 1871 

Horie, Yoshinori 2177 

Horii, Rika 1014 

Horiike, Norio 524 

Horner, Mary 1113, 1588, 1704, 

1744, 2053, 2054 

Hornsey, Emma 798 

Horsfall, Leigh 784 

Horsmans, Yves J. 1051 

Horvath, Angela 59, 2068 

Hosaka, Tetsuya 1062, 1191, 2020, 

2241 

Hoshida, Yohmei 513 

Hoshida, Yujin 229, 253, 254, 364, 

778, 1337, 1392, 1742, 1940, 2174 

Hoshino, Takashi 477 

Hosho, Keiko 1499 

Hosoya, Mitsuaki 551 

Hot, Edina 893 

Hou, Feng-Qin 2023 

Hou, Jin-Lin 2013 

Hou, Jun 1637 

Hou, Mengjun 454 

Hou, Ming-Chih 738, 751, 757, 759 

Hou, Xiaohua 1441 

Houben, Tom 922 

Houghton, David 2183 

Houlihan, Diarmaid D. 1103, 1216, 

1275 

Houry, Mohamad 2173, 2192 

House, Michael J. 798 

Houssel-Debry, Pauline 3, 95, 1781 

Housset, Chantal 196, 642, 786, 1409 

Hov, Johannes R. 833 

Howarth, Rachel M. 1381, 1408 

Howe, Anita Y. 40, 42, 210, 700, 701, 

703, 707 

Howell, Brett A. 224, 228 

Howieson, Kevin 1051 

Hsiang, John C 1612 

Hsieh, Matthew 1498 

Hsieh, Tsai-Yuan 1996 

Hsieh, Wei-Yao 751 

Hsieh, Yun-Cheng 757, 965 

Hsin, I-Fang 738, 751 

Hsing, Ann W. 207 

Hsu, Chia-Yang 365, 409 

Hsu, Hong-Yuan 1701, 1705 

Hsu, Ping-I 738 

Hsu, Shih-Jer 1996 

Hsu, Shu-hao 493 

Hsu, Yao-Chun 1547 

Hu, Bingqian 79 

Hu, Chengcheng 1716 

Hu, Chi-Tan 1554, 1996 

Hu, Donglei 78 

Hu, Haihong 1811, 1871 

Hu, Huaidong 1647 

Hu, Hui-Han 1593, 1604 

Hu, Jiangting 1925 

Hu, Jinhua 1675 

Hu, Junjie 1934 

Hu, Ke-Qin 85, 1031, 1165, 1785 

Hu, Peng 1044, 1647 

Hu, Richard 904 

Hu, Shi 1910 

Hu, Tsung-Hui 1873, 1942, 1996, 

2010 

Hu, Wei 100, 1924 

Hu, Xiaojun 1755 

Hu, Xudong 879, 1324, 1338 

Hu, Yaoren 2048 

Hu, Ying 508, 649, 674 

Hu, Yiran 1065 

Hua, Rui 1149, 2107 

Huaman, Moises 854 

Huang, Changxing 984 

Huang, Chao-Cheng 1942 

Huang, Chi-Wen 1701 

Huang, Chia-Lin 2040 

Huang, Chiung-Kuei 256, 1962 

Huang, Chung-Feng 2264 

Huang, Guangyu 1865 

Huang, Hai 184 

Huang, Hongfei 2024 

Huang, Jee-Fu 398, 2264 

Huang, Jhy-Shrian 398 

Huang, Kai-Wen 118, 1449 

Huang, Li 454 

Huang, Shu-Pang 720 

Huang, Shuo 1988 

Huang, Vivian 1606 

Huang, Wen 1812 

Huang, Xiaobi 731 

Huang, Xiaohui 1772 

Huang, Yi 1264 

Huang, Yi-Hsiang 365, 409, 439, 759, 

965, 1996, 2050 

Huang, Yi-Wen 1996 

Huang, Ying 1441 

Huang, Zhanlian 1423 

Huard, Genevieve 1997 

Huber, Caroline 1144, 1462 

Huber, Hans 118 

Hubers, Lowiek M. 637 

Hübner, Marc P. 2077 

Huck, Ian 650 

Hucke, Florian 475 

Huddleston, Heather 2252 

Huddleston, Leslie 1121 

Hudock, Rebecca 1570 

Hudson, Benjamin E. 564, 1140, 1173 

Hudson, Mark 2226 

Huebert, Robert C. 692, 1355 

Huelin, Patricia 266 

Hueppe, Dietrich 1060, 1078, 1081, 

1156, 1861 

Hughes, Dempsey 412 

Hughes, Dominic A. 1691 

Hughes, Eric A. 702 

Hughes, Michael 1094 

Hui, Aric Josun 241, 2014, 2025 

Hui, Peter 2104 

Hull, Katherine L. 62 

Hullegie, Sebastiaan 1007 

Hultcrantz, Rolf W. 158 

Hum, Dean W. 105, 162, 974, 2145 

Humar, Abhinav 838 

Humar, Bostjan 499, 665 

Humberson, Annette 1265 

Humbert, Lydie 816 

Hummels, Hazel 1373 

Humphrey, Mark 1986 

Hunault, Gilles 2188 

Hung, Annie K. 462 

Hung, Chao-Hung 1873, 2010 

Hung, Hsu-Wei 1547 

Hung Wan, Isabel 1121 

Hunt, Christine M. 540 

Hunt, Katie 1712 

Hunt, Kristel K. 16 

Hunt, Sharon L. 18, 315, 534, 1035, 

1460 

Hunter, Jill E. 652 

Hunter, Stuart 1298 

Huntsman, Scott 78 

Huntzicker, Erik G. 632, 1359, 2149 

Huo, Teh-Ia 356, 365, 409, 439, 759 

Huppert, Kari A. 680 

Huppert, Stacey S. 680 

Hur, Chin 154 

Hur, Keun 403, 426, 749 

Hur, Wonhee 1419 

Hurt, Christopher B. 1063 

Hussain, Shabir 581, 1344 

Hussain, Zilla H. 580 

Hussaini, Kamran 1535 

Hussaini, Tarana 204 

Hussein, Abd Elrazek A. 858, 2215 

Hutabarat, Renta M. 36 

Hutchinson, Sharon 1794 

Hutton, David W. 67 

Huynh, Andrew 1182, 1549 

Huynh, Dep K. 76 

Hwang, Jaeseok 2038 

Hwang, Kyu-Hee 1401 

Hwang, Peggy 42, 251, 700, 701, 

703, 715 

Hwang, Seong Gyu 1542 

Hwang, Sunil 1923 

Hyland, Robert H. 38, 247, 713, 777, 

1034, 1045, 1120, 1130, 1442, 1860


Hylemon, Phillip B. 22, 804, 1463, 

1521, 1527 

Hynan, Linda S. 1776 

Hyogo, Hideyuki 2156, 2158, 2199, 

2202, 2207 

Hyun, Jeongeun 696 

Hyun, Jinhee 1017 

Iannitti, David A. 653 

Iaria, Pierre 2082 

Iavarone, Massimo 229, 393, 435 

Ibanez, Luisa 225 

Ibis, Mehmet 932 

Ibrahim, Samar 895 

Ibusuki, Rie 937, 1421, 1838 

Ichai, Philippe 1237, 1269, 1782 

Ichikawa, Tatsuki 1671 

Ichiki, Yasunori 2031 

Ida, Kinuyo 697 

Ide, Tatsuya 304, 1448, 1819 

Ide, Yasushi 296 

Idilman, Ramazan 558, 1822, 2012, 

2033 

Ido, Akio 799, 937, 1421, 1838 

Idowu, Michael 905 

Ieluzzi, Donatella 1166 

Iemmolo, Rosa Maria 1751 

Iezzi, Roberto 432 

Iguchi, Kohta 402 

Iida, Noriho 370, 392, 430 

Iida, Takashi 1696 

Iijima, Hiroko 466, 770, 781, 1533, 

1669, 1975 

Iijima, Sayuki 1018 

Iimuro, Yuji 188 

Iio, Etsuko 1006, 1097, 1448, 1655, 

1954, 1983 

Iio, Sadaharu 1809 

Ijuin, Sho 799, 937, 1838 

Ikarashi, Yuichi 1326 

Ikeda, Fusao 2236 

Ikeda, Hiroki 1446 

Ikeda, Kazuo 517, 592 

Ikeda, Kenji 1062, 1191, 2020, 2241 

Ikeda, Naoto 466, 770, 781, 1533, 

1669, 1975 

Ikeda, Yasuhiro 2 

Ikeda, Yoshio 2146 

Ikegami, Tadashi 970 

Ikejima, Kenichi 226, 917, 1771 

Ikenaga, Naoki 826, 1367, 1368, 

1379 

Ikeno, Yoshinobu 1213 

Ikezono, Yu 688 

Ilan, Yaron 46 

Ilani, Nadav 1878 

Ildstad, Suzanne 54 

Iliaz, Raim 478 

Iliceto, Sabino 268 

Illa, Xavi 8 

Ilyas, Ghulam 1333 

Im, Gene Y. 1095, 1185, 1337 

Imagawa, Kazuo 1690 

Imai, Norihiro 880 

Imai, Yasuharu 1195, 1197, 2213 

Imai, Yukinori 1474, 1486 

Imai, Yumi 807 

Imai, Yusuke 338, 788, 1293 

Imajo, Kento 2207 

Imam, Mohamad 76 

Imamine, Rinpei 402 

Imamura, Masatoshi 794, 1795, 2158, 

2193 

Imamura, Michio 472, 986, 1056, 

1150, 1627, 1645, 1649, 1670, 

1681, 1858, 1893, 1895, 1944, 2120 

Imazeki, Fumio 1207 

Imbert-Bismut, Françoise 786, 1510 

Imteyaz, Hejab 280 

Inada, Masami 1197, 1201 

Inada, Yuki 370 

Inagaki, Yutaka 1369 

Inami, Yoshihiro 1771 

Inao, Mie 1059, 1064, 1474, 1486, 

2241 

Iñarrairaegui, Mercedes 344, 378 

Indenbirken, Daniela S. 1629 

Ingiliz, Patrick 1058, 1060, 1081, 

1156 

Inglot, Malgorzata 1161 

Ingviya, Thammasin 280, 294, 560 

Inomata, Kenta 336 

Inoue, Atsuo 173, 1201, 2213 

Inoue, Haruhiro 1941 

Inoue, Jun 1026, 1648, 1943, 2167 

Inoue, Taisuke 1565, 1673 

Inoue, Takako 1573, 1599, 1618 

Inoue, Tomoyoshi 756 

Intagliata, Nicolas 761 

Invernizzi, Federica 2011, 2012 

Invernizzi, Pietro 73, 76, 602, 609, 

621, 634, 1654 

Ioannou, George N. 49, 128, 230, 

540, 803, 923, 1211, 2151 

Iodice, Valentina 431 

Iovanescu, Vlad F. 755 

Ipharraguerre, Ignacio R. 194 

Iracheta-Vellve, Arvin 180, 496, 1318, 

1330, 2198 

Iredale, John P. 62, 260, 329, 753 

Ireland, Hamish 753 

Iriana, Sentia 566 

Irie, Makoto 385, 410 

Irvine, Katharine 784, 1362 

Irving, William 1140, 1173 

Isakov, Vasily 721 

Ishiba, Hiroshi 886, 888, 2230 

Ishibashi, Hiromi 2031 

Ishibashi, Naoto 657 

Ishida, Hisashi 1584 

Ishida, Kosuke 325, 947 

Ishida, Yuji 1670, 1679 

Ishigami, Masatoshi 411, 443, 880, 

967, 1061, 1167 

Ishihara, Akio 1584 

Ishii, Akio 466, 770, 781, 1533, 1669 

Ishii, Kunihide 304 

Ishii, Takamichi 1976 

Ishikawa, Tsuyoshi 760, 1447, 1464, 

1492 

Ishizu, Yoji 411, 443, 880, 967, 1061, 

1167 

Islam, Fakhar U. 1257 

Isogawa, Masanori 1640, 1655 

Isoyama, Shigemi 1690 

Issachar, Assaf 875 

Isted, Alexander 1733 

Itakura, Jun 1115, 1192 

Itami, Saori 429 

Itano, Osamu 336 

Ito, Daisaku 382 

Ito, Kiyoaki 2158 

Ito, Koichi 135 

Ito, Masahiko 1393 

Ito, Rei A. 1420 

Ito, Sayaka 505 

Ito, Takayoshi 1302, 1941 

Ito, Toshifumi 796, 1197 

Itoh, Fumio 1006, 1446 

Itoh, Ikuyo 901 

Itoh, Jun 287, 1946 

Itoh, Yoshito 232, 707, 886, 888, 901, 

1980, 2156, 2202, 2230 

Itokawa, Norio 291, 476 

Itou, Minoru 233 

Itsui, Yasuhiro 164, 406, 1018, 1027, 

1959 

Iwagami, Yoshifumi 256, 1962 

Iwaisako, Keiko 1918 

Iwakiri, Katsuhiko 291 

Iwakiri, Yasuko 1322, 1526 

Iwama, Itaru 1724 

Iwamoto, Hideki 688 

Iwamoto, Junichi 970 

Iwamoto, Marian 725, 730, 731 

Iwamoto, Takuya 760, 1464, 1492 

Iwao, Masao 2267 

Iwasa, Motoh 584, 1500 

Iwasaki, Ryuichiro 1584 

Iwasaki, Tetsuya 1584 

Iwase, Tomoya 1618 

Iwata, Kaoru 410 

Iwata, Tomoaki 2167 

Iwata, Yoshinori 466, 770, 781, 1533, 

1669, 1975 

Iwelu, Chibuzor 1233 

Iyer, Malliga 1374 

Iyer, Radhakrishnan P. 2262 

Izmaylov, Michelle 1223 

Izumi, Kousuke 1771 

Izumi, Namiki 1115, 1192, 1632, 

2029 

Izumi, Takaaki 1672 

Izzo, Francesco 851 

J Sangwaiya, Minal 791, 800 

Jaber, Fadi L. 61, 340 

Jablkowski, Maciej S. 1678 

Jachymova, Marie 1487 

Jackson, Kathryn 563, 849 

Jackson, Kathy 1566 

Jacobs, W. Carl 1023 

Jacobson, Ira M. 42, 91, 1086, 1106, 

1841 

Jacobson, Karen B. 89 

Jacomet, Christine 1093 

Jacomino, Kristina 1223 

Jacquemin, Emmanuel 196 

Jacquemyn, Bert 39 

Jacques, Vincent 143 

Jadeja, Ravirajsinh 1911 

Jadhav, Vasant 36 

Jaeschke, Hartmut 585, 586, 598, 

1767, 1774, 1925 

Jafoui, Sami 1779 

Jafri, Syed M. 307, 1872 

Jahnel, Jörg 1726 

Jain, Mamta K. 438 

Jain, Surbhi 1554 

Jain, Vikas 1340 

Jalan, Rajiv 113, 908, 1300, 1482, 

1517, 1769, 1780, 1781, 2069


Jalan-Sakrikar, Nidhi 692 

James, Laura 99, 1763, 1914 

James, Spencer L. 580 

James, Theodore W. 1255 

Jamil, Khurram 278 

Jamwal, Kapil D. 2070, 2092 

Janardhan, Sujit V. 522 

Jang, Byoung Kuk 2038 

Jang, Eun Chul 924, 941, 1760, 2110, 

2218, 2243 

Jang, Eun Sun 389, 2113 

Jang, Jae Seong 857 

Jang, Jae Yool 857 

Jang, Jae Young 277, 484, 736 

Jang, Jeong Won 452, 1615, 1882 

Jang, Kiseok 2218 

Jang, Se Young 363, 403, 426, 749 

Jang, Seung Hyeon 2049 

Jang, Sun Kyung 363, 403, 426, 749 

Janjua, Naveed Z. 86, 543 

Jansen, Christian 773 

Jansen, Geraldine 1613 

Jansen, Jos 2102 

Jansen, Louis 1633, 1995, 2005, 2046 

Jansen, Peter L. 604 

Janssen, Harry L. 73, 121, 241, 243, 

348, 350, 445, 542, 616, 634, 1099, 

1166, 1556, 1585, 1586, 1590, 

1637, 1651, 1998, 2001, 2002, 

2003, 2012, 2013, 2037, 2059, 2255 

Janssen, Katrien 1040 

Jara, Evelyn 971 

Jarnik, Michal 20 

Jaroszewicz, Jerzy 1502 

Jarufe, Nicolás 1217 

Jarupongprapa, Saharat 467 

Jaruvongvanich, Veeravich 964 

Jaskowski, Lesley-Anne 975 

Jasser-Nitsche, Hildegard 1726 

Jauffret-Roustide, Marie 1845 

Javaherian, Kavon 1451 

Javaid, Amen 286 

Javaid, Asad 371 

Javle, Milind 130, 682 

Jayaprakash, Aravindakshan 118 

Jayasekera, Channa R. 199, 201, 541, 

1534, 1745, 1750 

Jeamsripong, Woramon 1631, 1662 

Jeddari, Safaa 165 

Jeen, Yoon Tae 1504 

Jeffers, Lennox J. 2256 

Jeffers, Thomas 956 

Jeffery, Hannah C. 1298 

Jeffrey, Gary P. 848, 1077, 1264, 

2099, 2185 

Jelsing, Jacob 978 

Jen, Chin-Lan 1593, 1604 

Jen, Kuang-Yu 1736 

Jenab, Mazda 170 

Jeng, Rachel Wen-Juei 1552 

Jenkins, Nancy 234 

Jenkins, Stephen 329 

Jensen, Donald M. 539, 544 

Jensen, Majken K. 1702, 2228 

Jeon, Christie 2083 

Jeon, Hoonbae 1266 

Jeon, Tae Joo 464 

Jeon, Woo Kyu 1624, 1992 

Jeong, Jong-Min 910, 1358 

Jeong, Sook-Hyang 389, 2113 

Jeong, Soung Won 277, 484, 736 

Jeong, Won-IL 910, 1358, 1414 

Jeong, Woo Kyoung 485 

Jeong, Yun Seong 130, 682 

Jesse, Michelle T. 556 

Jessen, Niels 301 

Jessop, Amy B. 547 

Jeurissen, Mike 922 

Jewell, Mark 66, 936, 1386 

Jezequel, Caroline 264 

Ji, Cheng 588 

Ji, Dong 1536, 1537, 2008 

Jia, Jidong 374, 1404, 1660, 2048 

Jia, Zhansheng 984, 1004 

Jiang, An 255 

Jiang, Deyuan 1076, 1860 

Jiang, Guanglong 1332 

Jiang, Hai-Xing 1465 

Jiang, Hong Xiu 209 

Jiang, Hui 911 

Jiang, Jie 1583 

Jiang, Jing 1149, 1193, 2018 

Jiang, Joy 1285 

Jiang, Lijuan 2257, 2259, 2260 

Jiang, Tao 1149 

Jiang, Wei 1383 

Jiang, Xuemin 2259, 2260, 2261 

Jiang, Yanjun 194, 1689 

Jiang, Yong 1544 

Jiang, Zhaoshi 1651, 1668 

Jiang, Zhenghui G. 2227, 2228 

Jiang, Zhiwei 729 

Jiao, Chunhua 1521 

Jiao, Jingjing 1370, 1424 

Jibara, Ghalib 851 

Jideh, Bilel 1450 

Jimbei, Paulina 31 

Jiménez, Wladimiro 65, 659 

Jimenez Exposito, Maria Jesus 37, 252, 

1058 

Jiménez-Castro, Mónica B. 342 

Jimeno, Carlota 1863 

Jin, Dongfang 1579, 1617 

Jin, Jinglan 1657 

Jin, Li 1541, 1579, 1617, 1773 

Jin, Michelle Q. 367, 388, 1848, 2009 

Jin, Mingjuan 359, 367, 388, 459, 

1806, 1816, 1848 

Jin, Quan 919 

Jin, Ran 2232 

Jin, Young-Joo 1542 

Jindal, Ankur 146, 2070 

Jindal, Rohit 25 

Jinushi, Masahisa 505 

Jirsa, Milan 787 

Jitraruch, Suttiruk 1711 

Jittorntam, Paisan 1999 

Jogasuria, Alvin 879, 1324, 1338 

Jogunoori, Wilma S 130, 2118 

Joh, Jae Won 400 

Joh, Takashi 1954 

John, Anil 1843 

John, Nimy 1037, 1074, 1075 

Johnson, Ann D. 2052 

Johnson, Casey 57, 1522, 1971 

Johnson, Courtney N. 2022 

Johnson, Daniel C. 152 

Johnson, Heather 648, 1741, 1754 

Johnson, Lynt B. 2118 

Johnson, Matt 579 

Johnson, Nirah 1801 

Johnson, Phillip 344, 489, 490, 851 

Johnson, Rena D. 521 

Johnson, Scott J. 1087, 1143 

Johnson, Sindhu 1713 

Jokelainen, Kalle 633 

Jonas, Maureen M. 136 

Jonas, Sven 1222 

Jones, Christopher T. 2257 

Jones, David 601, 611, 625, 631, 636, 

1457, 2226 

Jones, Dean P. 19, 2232 

Jones, DeAnn 1174 

Jones, Elizabeth C. 2144 

Jones, Fiona 1321 

Jones, Hannah 822 

Jones, Rachel E. 1114 

Jones, Robert M. 1225 

Jong, Simcha 1455 

Joo, Sae Kyung 2211 

Joo, Seung-Moon 464 

Jopson, Laura 611 

Jordan, Cynthia E. 1792 

Jorge-Ripper, Carlos 1341, 1342 

Josbeno, Deborah A. 1244 

Joseph, John 2099 

Joshi, Shobha 1242 

Joshi, Y.K 1340, 2088 

Joshi-Barve, Swati 1304, 1331 

Jouannaud, Vincent 2082 

Jourdan, Tony 1374 

Joy, Jeffrey B. 1832 

Joyce, Andrew R. 1309 

Ju, Hye Lim 504 

Juday, Timothy R. 1042, 1068, 1069, 

1087, 1139, 1143, 1144, 1152, 

1198, 1462 

Julian, Schulze zur Wiesch 1629 

Julich, Henrike 43, 1299, 1385 

Jun, Baek Gyu 484 

Jun, Dae Won 924, 941, 951, 1760, 

2110, 2218, 2243 

Jun, Mi-Jung 446 

Jung, Minoa K. 2174 

Jung, Yong Jin 685, 1277 

Jung, Young Kul 391, 405, 1504, 1897 

Jung, Youngmi 696 

Jüngst, Christoph 617 

Juran, Brian D. 76, 600, 607, 610 

Jurek, Marzena 1467, 1478, 1485 

Justice, Amy 1488 

Kaasis, Mehdi 264 

Kabe, Yasuaki 1936 

Kabiri, Mina 104, 151 

Kachaylo, Ekaterina 665 

Kachuwaire, Obert 1575 

Kaczmarek, Dominik J. 1287, 2077 

Kado, Akira 912 

Kadry, Zakiyah 9 

Kaestner, Klaus H. 494, 511 

Kagawa, Tatehiro 314, 383, 697 

Kagay, Christopher R. 2104 

Kage, Masayoshi 1690 

Kahn, Michael 1945 

Kai, Yugo 97, 231, 669, 1082, 1357, 

1661, 1985, 2130, 2136 

Kaido, Toshimi 1213, 1935 

Kaiser, Ralf 2086 

Kaiser, Tiffany E. 873 

Kaita, Kelly D. 243


Kajanachumpol, Saowanee 1999 

Kaji, Kiichiro 370 

Kaji, Kosuke 677 

Kajihara, Mikio 2074 

Kajiwara, Atsushi 1941 

Kajiwara, Keiko 1375 

Kakati, Bobby 1043, 1109, 1224 

Kakati, Donny 735, 1220, 1253 

Kakazu, Eiji 58 

Kakinuma, Sei 164, 406, 1018, 1027, 

1959 

Kakisaka, Keisuke 1528 

Kakiyama, Genta 804 

Kakizaki, Satoru 477 

Kakolyris, Stylianos 1900 

Kakuda, Thomas 39 

Kakuda, Yuko 820 

Kalabin, Aleksandr 1282 

Kalafateli, Maria 2094, 2095 

Kalal, Chethan 1340 

Kalathil, Sumodh 1121 

Kalergis, Alexis 971 

Kalia, Harmit S. 943 

Kaliamoorthy, Ilankumaran 859 

Kalinina, Iryna 2162, 2186 

Kalkan, Cagdas 2033 

Kallail, K. James 1814, 1815 

Kallwitz, Eric R. 1248 

Kalmeijer, Ronald 1040 

Kalpakou, Georgia 2096 

Kalra, Avash 2085 

Kalra, Naveen 214 

Kalsekar, Anupama 711, 1456, 1461 

Kalthoff, Sandra 1927, 2108 

Kalwaney, Shirley K. 575 

Kamachi, Hirofumi 866 

Kamada, Yoshihiro 227, 906, 1642, 

1974, 2156, 2207 

Kamar, Nassim 3, 1158 

Kamath, Binita M. 1694, 1713 

Kamath, Patrick S. 6, 82, 281, 282, 

293, 570, 1315, 1316, 2089 

Kamble, Pravin S. 1050 

Kamel, Yasser M. 1147, 1188 

Kamimura, Hiroteru 310 

Kamiya, Akihide 164, 697, 1369 

Kamiya, Kenji 551 

Kamiyama, Toshiya 866 

Kampa, Nicholas 1718 

Kan, Hiromi 986, 1056, 1150, 1649, 

1670, 1681, 1858, 1895, 1944, 2120 

Kanai, Takanori 181, 569, 2131, 

2140, 2234 

Kanai, Tomoya 2074 

Kanda, Tatsuo 1207, 1628 

Kane, Pauline A. 212 

Kaneko, Akira 1201, 2213 

Kaneko, Shuichi 144, 168, 325, 370, 

392, 430, 495, 699, 947, 990, 994, 

1000, 1010, 1014, 1348, 1377, 

1395, 1632, 1653, 1953, 1968, 1970 

Kaneko, Shun 164, 406, 1959 

Kaneko, Syun 1018 

Kanel, Gary C. 1350 

Kanemasa, Kazuyuki 2230 

Kaneyama, Yasunari 2097 

Kang, Ah-Young 464 

Kang, Chaeryon 1598 

Kang, Dae Joong 1521 

Kang, Daehwan 876 

Kang, Jong-Hon 1097, 1778 

Kang, Ningling 1947 

Kang, Ray 71 

Kang, Seong Hee 405, 1897, 2017 

Kang, SoHee 1415, 1972 

Kang, Sun Woo Sophie 1909 

Kani, Satomi 1618 

Kanna, Sowjanya 1827 

Kantarjian, Hagop 1160 

Kanto, Tatsuya 476, 794, 1138, 1632, 

1643, 1658, 1795, 2158, 2193 

Kanwal, Fasiha 90, 104, 151, 244, 

357, 1048 

Kanwar, Bittoo 632, 1707 

Kao, Chia-Ning 2252 

Kao, Jia-Horng 1080, 1105, 1547, 

1828, 1996, 2027 

Kao, Shih-Chu 2258 

Kapalko, Angela 1136 

Kapeller, Rosana 957, 1938 

Kaplan, David E. 16, 122, 438, 521, 

1041, 1302, 1608 

Kaplan, Keith 1023 

Kaplan, Lee M. 154 

Kaplowitz, Neil 590, 1350, 1922, 

2135 

Kaps, Leonard 1353, 1426 

Kapur, Saurabh 748, 769 

Kar, Premashis 1596 

Karaca, Cetin 478 

Karagozian, Raffi 1469 

Karakan, Tarkan 932 

Karakaya, Fatih 2033 

Karani, John B. 212 

Karasu, Zeki 246, 558, 1247, 1739, 

1822 

Karatapanis, Stylianos 1126 

Karatayli, Ersin 2033 

Karatayli, Senem C. 2033 

Kardashian, Ani A. 1273 

Karimova, Madina 1629 

Karimzadeh, Hadi 31, 1021 

Karin, Michael 232, 1971 

Karino, Yoshiyasu 707, 1116, 1655, 

1778, 2065 

Karkada, Joel G. 1099 

Karlsen, Tom H. 76, 258, 819, 833 

Karnik, Satyajit 632, 1359, 1379, 2149 

Karnsakul, Wikrom 280, 294, 560 

Karp, Seth J. 837 

Karpen, Saul J. 19, 133, 1694 

Karpi, Asia 1885, 1886 

Karra, Vijay 1596 

Karrar, Azza 316, 2173, 2192 

Karsdal, Morten A. 119, 441, 780, 

1432, 1437, 2178 

Karthikeyan, Palaniswamy 1691 

Kartoun, Uri 2194 

Karvellas, Constantine J. 1759 

Kasai, Yosuke 402 

Kaseb, Ahmed O. 1160 

Kaser, Arthur 258 

Kashyap, Amit 1459 

Kasirga, Erhun 1623 

Kaspar, Mathew 1840 

Kasperkovitz, Pia 36 

Kassa, Anketse D. 1945 

Kasumov, Takhar 330, 651, 667 

Katayama, Hokahiro 1976 

Katayama, Kazuhiro 1201 

Kathpalia, Priya 557 

Katlama, Christine 210 

Kato, Akinobu 1528 

Kato, Junji 1984 

Kato, Naoya 505 

Kato, Takanobu 985, 1003 

Kato, Tomoaki 2129 

Kato, Yojiro 2129 

Kattakuzhy, Sarah 92, 220 

Kattamis, Antonios 1126 

Kattentidt-Mouravieva, Anja A. 492 

Katz, Barry P. 1316 

Katz, Jason 527 

Katz, Lior 2118 

Kaur, Savneet 954 

Kautiainen, Hannu 633 

Kawabata, Kenji 1690 

Kawada, Norifumi 191, 429, 517, 592, 

707, 1125, 1420, 1748, 1824, 2156 

Kawaguchi, Kazunori 168, 1653 

Kawaguchi, Takumi 233, 918, 1819, 

2158, 2207 

Kawaguchi, Yasunori 296 

Kawai, Takayuki 1976 

Kawai-Kitahata, Fukiko 164, 406, 1018, 

1027, 1959 

Kawakami, Yoshiiku 472, 986, 1056, 

1649, 1681, 1893, 1895, 1944 

Kawamoto, Hirofumi 2030, 2199 

Kawamura, Etsushi 1125, 1824 

Kawamura, Satoshi 382 

Kawamura, Yusuke 1062, 1191, 2020, 

2241 

Kawanaka, Miwa 2030, 2199, 2207 

Kawanishi, Koki 382 

Kawano, Akira 2031 

Kawaoka, Tomokazu 472, 986, 1056, 

1649, 1681, 1893, 1895, 1944 

Kawasaki, Yukihiko 551 

Kawashima, Keigo 1640 

Kawashima, Minae 1138 

Kawata, Kazuhito 1393 

Kawazoe, Seiji 296 

Kawut, Steven M. 9, 1470 

Kaya Kilic, Esra 2051 

Kayali, Zeid 702, 1100 

Kayandabila, Johnstone 572 

Kaymakoglu, Sabahattin 478, 558, 

1822 

Kazankov, Konstantin 119 

Kazerouni, Firouzeh 554 

Kazim, Syed N. 1005 

Kazimi, Marwan 1274 

Ke, Bibo 1283, 2138 

Ke, June 2261 

Keaveny, Andrew P. 1038, 1124 

Kebapcilar, Levent 2206 

Kedarisetty, Chandan K. 1328, 2078, 

2203 

Keefe, Deborah L. 2147, 2208 

Keegan, Mathew J. 1243 

Kegasawa, Tadashi 26, 453, 1809, 

1982 

Keiding, Susanne 768 

Keim, Sofia 1163 

Keirstead, Natalie 36 

Keller, Bradley T. 19 

Keller, Salome A. 1884 

Kelley, Caroline 711 

Kelley, Michael J. 540


Kelley, Robin K. 414, 471 

Kellogg, Todd A. 836 

Kellum, John A. 275 

Kelly, Catherine 931 

Kelly, Deirdre A. 1234, 1710, 1728 

Kelly, Dympna 850, 1271 

Kelly, Erin 1444, 1570 

Kelly, Mollie 33, 2064 

Kelly, Sean G. 434 

Kemgang Fankem, Astrid Donald D. 642 

Kemmer, Nyingi M. 526, 1240 

Kemp, William W. 442, 483 

Kempe, Allison 137, 1738 

Kemper, Sherri 192, 1356 

Kennedy, Lindsey 190, 621, 812, 822, 

835, 1317, 1363 

Kennedy, Patrick T. 167, 1571, 1626 

Kennedy, Susan M. 911, 1365 

Kenneth, Fleming A. 2190 

Kenny, Thomas P. 77 

Kent, Robert N. 1366 

Keppel, Jessica A. 1215 

Kerbert, Annarein J. 2069 

Kerbouche, Rafik 1621 

Kerkar, Nanda 133, 1694 

Kerlan, Robert 471 

Kersey, Kathryn 1076 

Kershaw, Erin E. 1531 

Kershenobich, David 2223 

Keskin, Onur 2012, 2033 

Keum, Bora 1438, 1504 

Khairy, Marwa 1076, 1163 

Khaitova, Viktoriya 1095, 1185 

Khalid, Hamza 1535 

Khalili, Korosh 348, 542 

Khalili, Mandana 306 

Khan, Aamir G. 1872 

Khan, Bilal 2047 

Khan, Harun 1826 

Khan, Mohammad Qasim 1791 

Khan, Rashid 281 

Khanam, Arshi 1762 

Khandelwal, Niranjan 214 

Khanna, Rajeev 1697, 1723 

Kharbanda, Kusum K. 589, 1323, 1347 

Khatri, Amit 1039, 1088 

Khattar, Ramzi 1012, 1297 

Khawaja, Shazia 717, 729 

Khawar, Sabrina 764 

Khemichian, Saro 217 

Khera-Butler, Tanya 113 

Khillan, Vikas 2070 

Khlaiphuengsin, Apichaya 2036 

Kho, Abel 563, 849 

Kholmukhamedov, Andaleb 1925 

Khoo, Mui Joo 1812 

Khoo, Saye H. 1103 

Khor, Seik-Soon 1138 

Khorzad, Rebeca 847 

Khosla, Ritu 695 

Khullar, Vikas 839 

Khungar, Vandana 864 

Khurana, Sandeep 1911 

Ki, Mo-ran 2113 

Kiciak, Slawomir 1582 

Kida, Akihiko 370 

KIda, Sachiho 906 

Kienle, Eike 98 

Kijsirichareanchai, Kunut 748, 769 

Kikuchi, Luciana 48, 380, 438, 457, 

469 

Kikuchi, Masahiro 2177 

Kikuchi, Miho 2177 

Kil, Natalie B. 1792 

Kilisinska, Natalia 1502 

Kim, Jung Hee 422, 485, 1960, 2049 

Kim, Ahyoung 1894 

Kim, Arthur Y. 1034, 1094 

Kim, Beom Kyung 346, 1542, 1558, 

1581 

Kim, Bo Hyun 377, 486, 2113 

Kim, Boo Sung 277, 484 

Kim, Boyoung 2116 

Kim, Brian 1151 

Kim, Byung Ik 1624, 1992 

Kim, Byung Seok 2038 

Kim, Chang Duck 1504 

Kim, Chang Wook 1542 

Kim, Chang-Min 486 

Kim, Chunki 920, 925 

Kim, Connie 272 

Kim, Dayoung 504 

Kim, Do Young 346, 504, 1542, 1558, 

1581 

Kim, Dong Joon 736, 1515 

Kim, Donghee 88, 1208, 2155 

Kim, Eui Joo 1896 

Kim, Eun Sun 1504 

Kim, Grace 272, 1454 

Kim, Gyeonghwa 403, 426, 749 

Kim, Haeryoung 1902 

Kim, Hee Yeon 1542 

Kim, Hong Joo 1624, 1992 

Kim, Hong Soo 277, 484 

Kim, Hwa Jung 2155 

Kim, Hwi Young 1277 

Kim, Hyeyoung 1277 

Kim, Hyo Jin 2049 

Kim, Hyun Joo 1352 

Kim, Hyung-Don 446 

Kim, Hyungwook 876 

Kim, Irene K. 1479 

Kim, Ja Kyung 464, 1399 

Kim, Jae Keun 464 

Kim, Jae-Sung 177 

Kim, Jeong Min 2038 

Kim, Ji Hoon 391, 405, 1504, 1897, 

2017 

Kim, Ji Young 577 

Kim, Ji-Hee 1401 

Kim, Jieun 696 

Kim, Jihoon 112 

Kim, Jin-Wook 389 

Kim, Jong Man 400 

Kim, Jonghwa 1415, 1972 

Kim, Ju Hyun 391, 1896 

Kim, Jun Won 464 

Kim, Jung-Hee 502, 1419 

Kim, Kang Mo 446, 448 

Kim, Karen E. 579 

Kim, Kyoungmi 606 

Kim, Kyung Hee 486 

Kim, Kyung-ah 2113 

Kim, Kyunga 422 

Kim, Kyungpil 241, 1678, 2004, 2043, 

2059 

Kim, Mi Na 1605 

Kim, Miran 256, 1962 

Kim, Misung 2227 

Kim, Moon Young 736, 1322, 1515, 

2080 

Kim, Myung-Ho 1358 

Kim, Nathan G. 387, 428 

Kim, Rosa S. 1392 

Kim, Sang 988, 1289 

Kim, Sang Geon 1352 

Kim, Sang Gyune 277, 484, 736, 

1542, 1605 

Kim, Sangbae 2118 

Kim, Seong Hoon 400, 862 

Kim, Seong-Jun 968 

Kim, Seung Up 346, 363, 1542, 1558, 

1581, 1605 

Kim, Shin Hee 277 

Kim, So Yeon 910, 1358 

Kim, Soo-Jin 1401 

Kim, Soon Sun 379, 1567, 2056 

Kim, Sung Min 1419 

Kim, Sunmi 277 

Kim, Tae Hyun 1352 

Kim, Tae Hyung 433, 1438, 1504 

Kim, Tae Suk 405, 1897, 2017 

Kim, Tae Yeob 736 

Kim, W. Ray 6, 75, 88, 381, 415, 546, 

562, 1182, 1189, 1208, 1813, 2155 

Kim, Won 1277, 2211 

Kim, Yong Ook 183, 950, 955, 1353, 

1376, 1385 

Kim, Yoon Jun 242, 400, 447, 470, 

474, 479, 1610, 1907, 2016, 2042, 

2055 

Kim, Yoona 207 

Kim, Young Don 277, 484, 1515 

Kim, Young Seok 277, 484, 736, 1542, 

1605 

Kim, Yun Ju 218 

Kim, Yun Soo 391, 1896 

Kim-Schluger, Leona 1185 

Kimura, Akihiko 1696 

Kimura, Kiminori 476 

Kimura, Koichi 1983 

Kimura, Minoru 1369 

Kimura, Mutuumi 1116 

Kimura, Naruhiro 310 

Kimura, Osamu 2167 

Kimura, Shoko 338 

King, Jennifer 708 

King, Jesse A. 630 

Kini-Matondo, Willy 1102 

Kinikli, Sami 2051 

Kinoshita, Manabu 324 

Kinoshita, Masahiko 1748 

Kipp, Benjamin R. 169 

Kiraithe, Muthamia 1666 

Kirby, Brian 1128, 1707 

Kirchner, Gabi I. 76 

Kirkness, Carmen S. 774 

Kirkpatrick, James N. 1218 

Kirkpatrick, Robert B. 874, 1260, 1280 

Kirpich, Irina 114 

Kirschner, Janina 1002 

Kirstein, Martha 444 

Kiser, Jennifer 1094 

Kishi, Hiroyuki 1599 

Kishino, Kyohei 466, 770, 781, 1533, 

1669 

Kisiel, John B. 169 

Kiso, Shinichi 227, 1974 

Kisseleva, Tatiana 186, 187, 189, 890


Kissiedu, Juliana 840 

Kita, Sadahiko 1976 

Kitade, Mitsuteru 689 

Kitagawa, Noriyuki 869 

Kitagawa, Yuko 336 

Kitago, Minoru 336 

Kitahara, Masaaki 370, 392 

Kitamura, Kazuya 1348 

Kitamura, Naoto 1978, 2143 

Kitazono, Masaki 456 

Kitrinos, Kathryn M. 243, 1551, 1557, 

1563, 1594, 1678, 2004, 2021 

Kitsahawong, Bubpha 555 

Kittitrakul, Chatporn 555 

Kitzman, Geoffrey 1178 

Kiyono, Soichiro 1490, 1516 

Kiyota, Ryousuke 1584 

Kjærgaard, Maria 773 

Klaassen, Curtis 655 

Klair, Jagpal S. 1099 

Klar, Ernst 1227 

Klarenbeek, Paul L. 637 

Klassen, Carolyn 647 

Klatte, Ryan 840 

Klebanoff, Matthew 154 

Klee, Eric W. 1965 

Klein, Andrew S. 1479 

Klein, Niklas 43, 1299, 1385 

Klein, Sabine 80, 961 

Klein, Thomas 955, 1376 

Kleiner, David E. 157, 166, 218, 597, 

779, 1498, 1646, 1717, 1720, 1722, 

1923, 1926, 2144, 2150, 2220 

Kleinewietfeld, Markus 1731 

Kleinstein, Sarah E. 2037 

Klenerman, Paul 1298 

Klepper, Arielle L. 229, 988, 1289 

Kliethermes, Stephanie 532, 559 

Klinck, John R. 1216 

Klinker, Hartwig 252, 1058, 1559 

Klintman, Daniel 308 

Klopfer, Stephanie 210, 712 

Klumpp, Klaus 33, 2064 

Kluwe, Johannes 2195 

Kneiber, David 57 

Knight, Virginia H. 47, 442, 483 

Knighton, Sarah 646, 1113, 1744 

Knisely, A. S. 1721 

Knoespel, Fanny 1921 

Knöpfli, Marina 425 

Knox, Steven J. 91, 1076, 1080, 1105, 

1120, 1168 

Knudsen, Sanne M. 978 

Ko, Eun Jeong 377 

Koay, Evelyn S. 1812 

Kobashi, Gen 551 

Kobayashi, Akira 869 

Kobayashi, Ichizo 1809 

Kobayashi, Mariko 1062, 1191, 2020 

Kobayashi, Masahiro 364, 1062, 1191, 

2020, 2241 

Kobayashi, Tomoki 472, 1056, 1893 

Kobayashi, Yoshimasa 1393 

Kobayashi, Yoshinao 584, 1500 

Kobayashi, Yoshiyuki 1047 

Kobazi Ensari, Gokce 831, 2115 

Kobune, Masayoshi 1984 

Koc, Özgür M. 1613 

Koch, Sandra 376 

Kocher, Hemant 101 

Koda, Masahiko 1499, 2199 

Kodali, Sudha 735 

Kodama, Kazuhisa 1326 

Kodama, Nobuhiro 2097 

Kodama, Takahiro 231, 234, 669 

Kodys, Karen 109, 180, 496, 512, 

1016, 2127, 2198 

Koek, Ger H. 922, 1613 

Koelfat, Kiran V. 604 

Koenecke, Christian 2128 

Koenig, Aaron B. 891, 2212 

Koga, Hironori 688, 918, 1819, 1962, 

1990 

Kogiso, Tomomi 1326, 2207 

Kogure, Takayuki 1026, 1648, 1943, 

2167 

Koh, Anna 364, 2174 

Koh, Christopher 218, 779, 1498, 

1650, 1717, 1720, 2144 

Koh, Hong 195 

Koh, Kwang Cheol 422, 423, 485, 

1591, 1960, 2049 

Koh, Sarene 167 

Kohgo, Satoru 1655 

Kohgo, Yutaka 927 

Kohjima, Motoyuki 1157, 1159 

Kohli, Anita 92, 220, 1111, 1155, 

1164 

Kohli, Rohit 159, 160, 916 

Koido, Shigeo 2074 

Koike, Kazuhiko 461, 912, 1138, 1632 

Koike, Kazuko 339, 2236 

Koizumi, Jun 383 

Koizumi, Wasaburo 482, 756 

Koizumi, Yohei 788, 1293 

Kojima, Kentaro 382 

Kojima, Sei-ichiro 383 

Kojima, Soichi 657 

Kokordelis, Pavlos 1287 

Koksal, Aydin 846 

Koksal, Iftihar 2057 

Kolachala, Vasantha L. 12 

Kolb Nava, Casey M. 1831 

Kolbeck, Kenneth J. 481 

Kolli, K. Pallav 471 

Kolly, Philippe 425 

Koloda, Dmitry 721 

Komar, Michael 552 

Komatsu, Nobutoshi 1565 

Komolmit, Piyawat 360, 1631, 1662 

Komori, Atsumasa 408, 783, 1600 

Komorizono, Yasuji 456 

Komuta, Mina 1734 

Kon, Kazuyoshi 226, 917, 1771 

Kondili, Loreta A. 1802, 1844 

Kondo, Chisa 291 

Kondo, Hisayoshi 952 

Kondo, Mario 1484, 1494, 1495 

Kondo, Mayuko 461 

Kondo, Mitsumasa 1970 

Kondo, Reiichiro 465 

Kondo, Takayuki 1490, 1516 

Kondo, Yasuteru 1026, 1648, 1943, 

1983, 2167 

Kondo, Yuichi 1420 

Kondo, Yuji 461 

Konerman, Monica 576 

Koneru, Alaya 1787 

Kong, Bo 919, 934 

Kong, Fei 1149, 2018 

Kong, Wenli 2107 

Kong, Yuanyuan 374 

Kongsomgan, Anucha 1999 

Konidari, Anastasia 1702 

Konijn, Cynthia 68 

Koning, Ludi 1196 

Kono, Hiroshi 1956, 2125 

Konsten, Sarah 1613 

Kontos, Georgios 2019 

Koo, Imhoi 926 

Koola, Jejo D. 529 

Koorey, David J. 1243 

Koot, Maarten 1995 

Kootstra, Neeltje A. 1633, 2005, 2255, 

2263 

Kopanja, Dragana 1988 

Kopec, Anna K. 81 

Koral, Kelly 1979 

Korayem, Enas 733 

Korenaga, Keiko 794, 1795, 2158, 

2193 

Korenaga, Masaaki 794, 1138, 1658, 

1795, 2158, 2193 

Korkmaz, Huseyin 2206 

Kormos Hallberg, Csilla 2100 

Kornek, Miroslaw 43 

Kort, Jens 41, 248, 250, 705, 710, 

719, 723 

Korzun, William J. 1776 

Kosaka, Hisashi 1420 

Koser, Martin 510, 2116 

Koskinas, Ioannis 1126 

Kosloski, Matthew P. 710, 723 

Kosters, Astrid 19 

Kostev, Karel 1052 

Kota, Janaiah 436 

Koteish, Ayman A. 607 

Kotoh, Kazuhiro 1159 

Kottilil, Shyam 92, 220, 578, 1013, 

1029 

Kou, Xiaoni 1773 

Koudou, Kazuki 901 

Kouides, Peter 1034 

Koukoulioti, Eleni 1638 

Kountouras, Dimitrios 1126 

Kourakli, Alexandra 1126 

Kourikou, Anastasia 2012, 2019 

Kouroumalis, Elias A. 2094 

Koury, Kenneth J. 700 

Koutroumpakis, Efstratios 2095 

Kouyama, Jun-ichi 1064 

Kouyos, Roger 1818 

Kouznetsova, Maria A. 528 

Kovacs, Andrea 1444 

Kovacs, Steven J. 2261 

Kowdley, Kris V. 73, 107, 239, 607, 

616, 628, 634, 946, 1046, 1106, 

1108, 1120, 2153, 2154, 2161, 2220 

Kowgier, Matthew 174, 348, 350, 

1099, 1556 

Koyama, Yukinori 189, 1522 

Koyanagi, Toshimasa 1159 

Kozak, Ladislav 622 

Kozbial, Karin 297, 732, 1070, 1092, 

1137, 1885 

Kozono, Masaya 937, 1421 

Kozuka, Ritsuzo 1125 

Kraft, Anke R. 2026 

Krag, Aleksander 119, 773, 780, 789, 

792, 1440


Krajden, Mel 86, 543 

Kramer, Jennifer R. 156, 244, 1048 

Krämer, Benjamin 1008, 1287, 2077 

Krams, Sheri M. 341, 2123 

Kranidioti, Hariklia 2019 

Krasnewich, Donna 1722 

Krastev, Zahary 2059 

Krattinger, Regina 519, 520 

Krauel, Alexander 245, 1594 

Kraus, Michael R. R. 1170, 1200 

Krawczyk, Marcin 43, 1429, 2195 

Krawczyk, Sarah 2227 

Kreefft, Kim 1007 

Krenkel, Oliver 1756 

Krenzien, Felix 1222 

Kresge, Charles 815 

Kretzschmar, Georg 617 

Kreutzfeltd, Martin 301 

Krishnamurthy, Venkat 418, 2116 

Krishnan, Anuradha 825, 1994 

Krisko, Tibor I. 881, 900 

Kriss, Michael 503 

Krittanawong, Chayakrit 282 

Krohn, Sandra 261, 276 

Krok, Karen 1470 

Krokowski, Dawid 668 

Kronborg, Ian 442 

Kronenberger, Bernd 1055 

Krones, Elisabeth 2068 

Kronmann, Karl 1175 

Krowka, Michael J. 9, 1470 

Kroy, Daniela C. 963 

Kruger, Annie 25 

Kruglov, Emma A. 661 

Krull, Mona 1708 

Ku, Hye Jin 346, 1558 

Ku, Lawrence 1185 

Ku, Winston 1182 

Kuate, Seraphin 200 

Kubik, Jacy L. 1969, 2139 

Kubo, Shoji 429, 1748, 1983 

Kuchimanchi, Satya 36 

Kuchipudi, Aishwarya 307 

Kucirka, Lauren M. 1749 

Küçükay, Fahrettin 846 

Kudo, Masatoshi 851, 1935 

Kudo, Yotaro 461 

Kuehne, Felicitas 1089 

Kugelmas, Marcelo 1040 

Kugiyama, Yuki 408, 783, 1600 

Kuiken, Sjoerd 1995 

Kuk, Nathan T. 945 

Kulkarni, Supriya 23 

Kullak-Ublick, Gerd A. 225, 519, 520 

Kuller, Lewis 2228 

Kumada, Hiromitsu 364, 707, 1062, 

1191, 2020, 2241 

Kumada, Takashi 344 

Kumagai, Erina 794, 1795, 2158, 

2193 

Kumagai, Kotaro 799, 1421, 1838 

Kumagai, Masashi 370 

Kumagai, Takanori 296 

Kumagi, Teru 524, 634 

Kumar, Aditi 1234 

Kumar, Ambuj 1240 

Kumar, Anupam 690, 694, 1483 

Kumar, Avinash 146, 262, 330, 334, 

518, 651, 667, 668, 671, 1308, 2088 

Kumar, Dhananjay 690, 694, 1483 

Kumar, Divya P. 807 

Kumar, Guresh 1328, 1340, 1344, 

1663, 2092, 2203 

Kumar, Manoj 1328, 1659, 2070, 

2203 

Kumar, Naveen 2245 

Kumar, Niteen 1268 

Kumar, Princy N. 1049 

Kumar, Sachin 2092 

Kumar, Vipin 1345 

Kumari, Radhka 1182, 1189 

Kumer, Sean C. 352, 565 

Kummen, Martin 833 

Kumral, Dennis 13 

Kunasegaran, Kamini 167 

Kuniholm, Mark H. 1444 

Kunimoto, Hideo 1191, 2020, 2241 

Kuno, Atsushi 1448 

Kunos, George 1374 

Kuo, Alexander 94, 1057 

Kuo, Margot E. 86, 543 

Kuo, Paul C. 2143 

Kuo, Yong-Fang 1249 

Kupiec-Weglinski, Jerzy 1283, 2138 

Kurahashi, Tomohide 227, 1974 

Kuramitsu, Tomoyuki 1097 

Kurimoto, Ami 1420 

Kurioka, Ayako 1298 

Kuroda, Hidekatsu 1528 

Kuroda, Shunichi 1642 

Kuromatsu, Ryoko 1819 

Kurosaki, Masayuki 1115, 1192, 1632, 

2029 

Kurosawa, Takao 1696 

Kurt, Ramazan 1096 

Kurt-Jones, Evelyn A. 180 

Kurtz, Tracie L. 138 

Kuruvilla, Asha 1513 

Kusakabe, Atsunori 1097 

Kusama, Hiromi 226, 917 

Kusano, Hironori 465, 1990 

Kusanovic, Juan P. 78 

Kuscuoglu, Deniz 2115 

Kushner, Tatyana 1608 

Kutay, Huban 99 

Kutsenko, Alina 359, 1806 

Kuwahara, Reiichiro 304, 681, 1819 

Kuwaki, Kenji 2236 

Kuwata, Yasuaki 1116 

Kuzuya, Teiji 411, 443, 880, 967, 

1061, 1167 

Kwak, Joo Hee 1760, 2110, 2243 

Kwanten, Wilhelmus J. 894, 976 

Kwee, Sandi 1553 

Kweon, Young Oh 363, 403, 426, 749, 

2028, 2038 

Kwo, Paul Y. 42, 217, 248, 700, 702, 

703, 712, 1084, 1903 

Kwok, Ryan M. 1101 

Kwon, Hyunjoo 686 

Kwon, Oh Sang 391, 1896 

Kwon, Sang Ok 2080 

Kwon, Seong Uk 857 

Kyritsi, Tina 190, 817, 1317 

Kyvernitakis, Andreas 1030, 1160 

L’Italien, Gilbert 1828 

La Rosa, Katia Yu 222 

Labadie, Hélène 264 

Labarga, Pablo 1881, 1908 

Labreuche, Julien 1269 

Labriola, Raffaella 983 

Labropoulou-Karatza C, Chrisoula 2094, 

2095 

Lacerda, Marco A. 1903 

LaCerte, Carl 1915 

Lachlan, Neil J. 260, 753 

Lacombe, Karine 1676, 1677, 1833 

Lacoste, Benoit 179 

LaCreta, Frank 720, 728 

Ladenheim, Maya R. 387, 428 

Ladner, Daniela 71, 563, 843, 847, 

849 

Laengvejkal, Pavis 280 

Lafdil, Fouad 693 

Laferl, Hermann 1885 

Lafoz, Erica 1518 

Lages, Celine S. 823, 1695 

Laggetta, Maristella 1493 

Lahiri, Pooja 831 

Lahoti, Manohar M. 1784 

Lai, Chengjung 510, 2116 

Lai, Ching-Lung 126, 1120, 1545, 

1550, 1746 

Lai, Hsueh-Chou 2040 

Lai, Jennifer B. 1053 

Lai, Jennifer C. 72, 557, 841, 844, 

852, 1219, 1468 

Lai, Jiaming 454 

Lai, Michelle 1484, 1494, 1495, 2160, 

2196, 2197, 2227, 2228 

Lai, Paul B. 344 

Lai, Sara 309 

Laitinen, Tarja 225 

Lakatos, Isobel 1707 

Lakdawalla, Darius 1042, 1068 

Lake, John R. 1184, 2006 

Lal, Bikrant B. 1697, 1725 

Laleman, Wim 171, 1091 

Lalezari, Jacob P. 41, 210, 1065 

Laliena, Almudena 1398 

Lalley-Chareczko, Linden 1136 

Lam, Angela 33, 2064 

Lam, Brian P. 892, 1186 

Lam, Christina 1722 

Lam, Hung 2175 

Lam, Karen S. L. 898 

Lam, Katherine 1100 

Lamb, Cheri L. 1403 

Lambiase, Lara 222 

Lamblin, Géraldine 1505 

Lamerato, Lois 1578 

Lammers, Willem J. 73, 601, 634 

Lammert, Frank 445, 463, 617, 1166, 

1429, 2077, 2086, 2195, 2253 

Lampe, Guy 784 

Lampertico, Pietro 1747, 2011, 2012 

Lan, Tian 890 

Landas, Steve 1961 

Landeen, Lee K. 1770 

Landen, Michiel 976 

Landini, Maria Paola 1888 

Landis, Charles S. 706, 716, 726, 

1049, 1211, 1478 

Landsittel, Doug 1244 

Lane, Suzanne 1139, 1152 

Lanford, Robert E. 32, 2022 

Lang, Anna L. 2141 

Lang, Julia 1666 

Lange, Undine G. 1222 

Langedijk, Jacqueline 827


Langhans, Bettina 1008, 2077 

Langhi, Cedric 65 

Langiewicz, Magda 499 

Langlet, Philippe 1091 

Langworthy, James 766 

Lankisch, Tim 1002 

Lanti, Claudia 2187 

Lanza, Marika 1654 

Lanzoni, Valéria P. 2189 

Lapalus, Martine 998, 1601, 2044, 

2045 

Lapierre, Pascal 305 

Lapinski, Tadeusz W. 1502 

Lapuyade, Bruno 450 

Laraque, Fabienne 1801 

Larrey, Dominique G. 206, 225, 1847 

Larsen, Lars P. 768 

Larsen, Lois M. 1107 

Larson, Colin 503 

Larson, Joseph J. 6 

Larsson Cohen, Marita S. 510, 2116 

LaRusso, Nicholas F. 73 

LaSalle, Janine M. 2109 

Lascoux-combe, Caroline 1676, 1677 

Laskin, Debra 934 

Lassailly, Guillaume 1269, 2222 

Lasser, Luc 1091 

Latinne, Dominique 1734 

Latour, Veronique 1796 

Lattanzi, Barbara 2067 

Latz, Eicke 100 

Lau, Audrey H. 2123 

Lau, Daryl 371 

Lau, George K. 1536, 1537, 2008 

Lau, Vincent 33, 2064 

Lauer, Georg 332 

Lauer, Ulrich M. 1183 

Lauletta, Gianfranco 435 

Laura, Alvarez 1919 

Laurent, Alexis 421, 449, 1232 

Lauridsen, Mette M. 1472 

Lauriski, Shannon 1849 

Lavanchy, Daniel 1818 

Lavert, Marion 999 

Lavieri, Mariel S. 67 

Lavine, Joel E. 159, 2161 

Lavrut, Pierre-Marrie 195 

Law, Matthew 1868 

Lawitz, Eric 39, 42, 205, 219, 247, 

700, 702, 703, 712, 718, 726, 1039, 

1040, 1120, 1128, 1428, 1433, 

1435, 2257, 2260 

Lawler, Joseph 1151 

Lawson, Mark 175, 915, 1916 

Layden, Thomas J. 1248 

Layese, Richard 362, 1808, 1847 

Lazaridis, Konstantinos 76, 600, 607, 

610 

Lazaro, Leander 1425 

Lazarus, Arnaud 1194 

Le, An K. 207, 359, 368, 387, 388, 

428, 1561, 1806, 1816, 1848 

Le, Michael H. 367, 459 

Le, Richard H. 359, 367, 386, 388, 

458, 459, 1806, 1816, 1848 

Le, Thuy T. 191, 1824 

Le Bail, Brigitte 1439, 2181 

Le Bert, Nina 167 

Le Cleach, Aline 1997 

Le Grand, Béatrice 30 

Le Hello, Rolland 1796 

Le Marchand, Chloe 1793 

Leaf, David 1488 

Leal, Carlos 2084 

Leal Tassias, Aránzazu 691 

Learned, Marc 198, 1986 

Leary, Thomas 1551 

Leathers, James 1223 

Leber, Bettina 2068 

Leber, Werner 1052 

Lebofsky, Margitta 586 

Lebray, Pascal 3, 95, 206, 786, 1239 

Lebrec, Didier 145, 265, 772, 1288, 

2148 

Lebreton, Aurelien 1505 

Lebrilla, Carlito B. 606 

Lebuffe, Gilles 1269 

Leclair, Katherine B. 881, 900 

Leclercq, Isabelle A. 178, 678 

LeCuyer, Brian E. 658 

Lee, Amy C. 2007, 2034, 2062 

Lee, Andy Hung-Yi 71, 847 

Lee, Brian P. 1214, 1215 

Lee, Chang Hee 1897 

Lee, Chang Hyeong 2038 

Lee, Christine K. 136 

Lee, Chuan-Mo 1873, 2010 

Lee, Danbi 446, 448 

Lee, David D. 50, 53, 335 

Lee, Donghyeon 242, 400, 447, 474, 

479, 1610, 2016, 2042, 2055 

Lee, Eun Byul 502, 1419 

Lee, Eung Chang 862 

Lee, Fa-Yauh 738, 751 

Lee, Guan Huei 1686 

Lee, Hae Lim 452, 1882 

Lee, Hae Won 1277 

Lee, Han Chu 446, 448 

Lee, Hannah 2015, 2052 

Lee, Helen 32, 2022 

Lee, Heon Ju 2038 

Lee, Hong Sik 1504 

Lee, Hye Won 1558, 1581 

Lee, Hyejung 1902 

Lee, I-Cheng 439 

Lee, Ik Jae 464 

Lee, Jai Sun 941 

Lee, Jeong Min 474 

Lee, Jeong-Hoon 242, 400, 447, 470, 

474, 479, 1610, 1907, 2016, 2042, 

2055 

Lee, Jin-Woo 1542 

Lee, Joon Ho 1419 

Lee, Joon Hyeok 422, 423, 485, 1591, 

1960, 2049 

Lee, Ju-Seog 129, 2118 

Lee, June Sung 242, 400, 447, 470, 

479, 1907, 2016 

Lee, Jung Il 464, 1399, 1542 

Lee, Kuei-Chuan 757, 965, 2050 

Lee, Kwan Sik 464, 1399, 1542 

Lee, Kwang-Hun 464 

Lee, Kwang-Woong 400, 474, 1277, 

1278 

Lee, Kwangwon 879, 1324, 1338 

Lee, Major 521 

Lee, Mei-Hsuan 1593, 1604, 1828 

Lee, Mikang 920, 925 

Lee, Minjong 242, 400, 447, 474, 479, 

1610, 2016, 2055 

Lee, Richard 78 

Lee, Sae Hwan 277, 484, 991, 1630, 

1644 

Lee, Samuel S. 1051, 1529 

Lee, Sangbin 436 

Lee, Sangmin 1336 

Lee, Serene M. 520 

Lee, Seulki 115 

Lee, Seung Duk 862 

Lee, Seung Min 924, 941, 1760, 2110, 

2218, 2243 

Lee, Soonkyu 452 

Lee, Sooyeon 177 

Lee, Su Hyun 363, 403, 426, 749 

Lee, Sunyoung 1783 

Lee, Taeheon 1624 

Lee, Teng-Yu 345 

Lee, Tingfang 1364 

Lee, Tzu-Hao 1887 

Lee, William M. 99, 211, 213, 1555, 

1759, 1761, 1763, 1766, 1768, 

1774, 1776, 1777, 1914 

Lee, Yoonkwang 920, 925 

Lee, Young-Sun 405, 1897, 2017 

Lee, Youngmin A. 1364 

Lee, Yu Rim 363, 403, 426, 749 

Lee, Yun Bin 2042, 2055 

Lee, Yung Sang 446, 448 

Leeflang, Mariska M. 74, 623 

Leeming, Diana J. 119, 441, 780, 1432 

Lefeber, Dirk 2102 

Lefebvre, Eric 1756, 2247 

Lefèvre, Mathieu 1019 

Leffondre, Karen 132 

Lefkowitch, Jay H. 2129 

Leggett, Barbara A. 106, 627 

Legrand, Margaux 1409 

Lehar, Sophie M. 2009 

Lehmann, Patrick 1002, 1572, 1597 

Lehretz, Maria 2012 

Leipertz, Steven 128 

Leise, Michael D. 1038, 1124 

Lemasson, Matthieu 997 

Lemasters, John J. 319, 1925 

Lemesch, Sandra 2068 

Lemmers, Arnaud 178 

Lemoinne, Sara 642 

Lemon, Stanley M. 990, 1010 

Lenci, Ilaria 203, 1751 

Lenders, Marie-Hélène 922 

Lenicek, Martin 604, 1487 

Lennon, Michael 1491 

Lenoci, Maryanne 1133 

Lens, Sabela 982 

Lenz, Oliver 1040, 1179 

Lenzen, Henrike 76 

Leonard, Jennifer 204 

Leong, Jennifer 1095, 1185 

Lepida, Antonia 2079 

Lerner, Daniel J. 531, 550 

Leroy, Vincent 3, 95, 117, 206, 777, 

1158, 1439, 1808, 1860 

Lesgourgues, Bruno 1072, 2082 

Letenneur, Luc 132 

Leteurtre, Emmanuelle 2222 

Letoublon, Christian 421, 449 

Letouze, Eric 362 

Leung, Patrick S. 77, 602, 606, 613, 

619, 639 

Levander, Sepideh 992


Leveille-Webster, Cynthia 582 

Leventhal, Thomas M. 1777 

Levesque, Eric 215, 1269 

Levi, David 17 

Levi, Moshe 139, 142, 907, 959 

Levin, Douglas M. 874, 1280 

Levine, Matthew H. 868 

Levine, Vanessa 731 

Levitsky, Josh 5, 54, 1235 

Levitt, Brian S. 1549 

Levrero, Massimo 165, 958 

Levstik, Alexander 2117 

Levy, Adrian 1152 

Levy, Cynthia 76, 616, 624, 1054, 

1148, 1206 

Levy, Gahl 331, 672, 679 

Levy, Gary 11, 424, 1012, 1297 

Lewis, David L. 32, 2022 

Lewis, Dylan 212 

Lewis, James D. 865 

Lewis, James H. 286 

Li, Lei 1934 

Li, Xiaojiao 2058 

Li, Biao 1668 

Li, Bing 1536, 1537, 2008 

Li, Changyong 1283, 2138 

Li, Chen 1609 

Li, Dongxiao 328 

Li, Fan 1536, 1537, 2008 

Li, Fang 2066 

Li, Feng 874 

Li, FengDI 1611 

Li, Fengyuan 1305 

Li, Guangyu 984 

Li, Haiyang 1315 

Li, Hu 1044 

Li, Hui 2048 

Li, Jia 496 

Li, Jiayi 207, 368, 1561 

Li, Jie 2018 

Li, Jin 1595 

Li, Jun 332 

Li, Kelvin 1425 

Li, Li 35, 1359 

Li, Liying 1396 

Li, Man 829 

Li, Mengsen 1977 

Li, Minran 2066 

Li, Qin 815 

Li, Qingmei 2058 

Li, Shuzhao 19, 2232 

Li, Tiangang 140 

Li, Tony 1952 

Li, Wanyu 1657 

Li, Wei 1977 

Li, Wenwen 505 

Li, Xia 87 

Li, Xiaodong 1595, 1675 

Li, Xiaolei 1948 

Li, Xiaoxia 675 

Li, Xin 1557 

Li, Xu 2048 

Li, Yingxia 942 

Li, Yinping 1869 

Li, Yong 1684 

Li, Yue 322 

Li, Yunzhou 22 

Li, Zhiping 1214, 1215 

Li, Zhongbin 1537, 2008 

Li, Zhuan 352, 1311 

LI, Jie 193, 197 

Lian, Jianqi 984 

Liang, Chen 2002 

Liang, Cheng-Chao 1996 

Liang, Guang 22 

Liang, Lijian 454 

Liang, Nathan L. 216 

Liang, T. Jake 218, 779 

Liang, Yan 1004 

Liangpunsakul, Suthat 267, 1316, 

1332, 1903 

Liao, Lijun 100 

Liao, Wei-Jia 1973 

Liaw, Yun -Fan 1552 

Liberal, Rodrigo 300, 608 

Lichvar, Alicia B. 1741, 1754 

Lidofsky, Anna 25, 991, 1015, 1630, 

1644 

Liedtke, Christian 828, 831, 1924 

Liedtke, Wolfgang B. 962, 2137 

Liefferinckx, Claire 178 

Liew, Zhong Hong 451 

Liffmann, Danielle 156, 1792, 1839 

Lifton, Richard P. 1719 

Lightbourne, Teisha G. 1451 

Lillegard, Joseph 2 

Lilley, Kirthi 420 

Lilly, Les 11, 204 

Lim, Arlene 1351 

Lim, Joseph K. 94, 540, 1057, 1488 

Lim, Kieron B. 1928 

Lim, Mary Ann 871 

Lim, Rebecca 945 

Lim, Sarah P. 47 

Lim, Seng Gee 241, 1686, 1928, 

2014, 2025 

Lim, Won 749, 876, 2028 

Lim, Yoo Li 2080 

Lim, Young-Suk 367, 446, 448, 459, 

1080, 1105 

Lima, Erika C. 2200 

Limani, Perparim 665 

Limbourg, Florian 2128 

Limothai, Umaporn 2036 

Lin, Andy 1850, 1877 

Lin, Chia-Hsin 2040 

Lin, Chih-Lin 1996 

Lin, Chih-Wei 41, 248, 250, 719 

Lin, Chih-Wen 398 

Lin, Chun-Che 1996 

Lin, Clement Y. 451 

Lin, Derek 207, 368, 386, 458, 1561 

Lin, Grace 1470 

Lin, Han-Chieh 365, 409, 439, 738, 

751, 757, 759, 965, 2050 

Lin, Henry C. 1693 

Lin, Jaw-Town 345, 1547, 1589 

Lin, Jianguo 1361 

Lin, Kai 2257 

Lin, Lanjia 243, 2025 

Lin, Lanyi 1611 

Lin, Linda I. 1264 

Lin, Lung-Huang 1701 

Lin, Ming 219, 1130 

Lin, Ming-Tsung 1873 

Lin, Ming-Tzung 1942 

Lin, Pey-Ru 1942 

Lin, Selena 1554 

Lin, Steven C. 2201 

Lin, Su 1765 

Lin, Wen-Terng 1701 

Lin, Wenyu 991, 1015, 1630, 1644 

Lin, Yong 1562 

Lin, Ziyu 1772 

Linaberry, Misti 702 

Linas, Benjamin P. 152, 1811 

Linda, Sher 200 

Lindell, Per 2258 

Lindhout, Darrin 1986 

Lindor, Keith D. 73, 75, 76, 546, 562, 

605, 630, 634 

Line, Pål-Dag 819 

Ling, John 1133, 1707 

Ling, Lei 106, 198, 627, 1986 

Ling, Victor 824 

Lingala, Shilpa 1717, 1722 

Lingiah, Vivek A. 295, 1181 

Linthicum, Mark T. 1042 

Liorre, Giulia 431 

Liou, Iris W. 1211 

Lippai, Dora 512 

Lippincott-Schwartz, Jennifer 1909 

Lisboa, Quelson C. 2200 

Lisman, Ton 740, 2249 

Lison, Hortensia 264, 554, 1072, 2082 

Lissock, Frédéric S. 1833 

Littera, Roberto 309 

Littlejohn, Margaret 1566 

Litwin, Alain H. 40 

Liu, Andy 199, 201, 1745, 1750 

Liu, Benny 7, 237, 1252, 2159 

Liu, Changlu 2147, 2208 

Liu, Chao 498, 1951 

Liu, Chen 138, 989, 1967 

Liu, Chen-Hua 1828 

Liu, Chun-Jen 2027 

Liu, Chunxiao 1537 

Liu, Fang 730, 731 

Liu, Fanwei 994, 1000 

Liu, Feng 782, 1436 

Liu, Fu-Tong 1285 

Liu, Hannah 1455 

Liu, HongLing 1609 

Liu, Hongqun 1529 

Liu, Hui-Xin 63, 508, 649, 674 

Liu, James 506 

Liu, Jessica 1593, 1604 

Liu, Jialiang 1536, 1537, 2008 

Liu, Jie 1374 

Liu, Jinfeng 1541, 1579, 1617, 1773 

Liu, Jingmei 328 

Liu, Ju 36 

Liu, KeHui 1611 

Liu, Lawrence 1095, 1185 

Liu, Lei 468 

Liu, Li 722 

Liu, Liming 1305, 1306 

Liu, Lin 247, 824, 1128 

Liu, Mengyuan 1266 

Liu, Miao 1562 

Liu, Po-Hong 365, 409 

Liu, Qian 1647 

Liu, Ran 41, 248, 1106 

Liu, Runping 22, 1521 

Liu, Sichen 807 

Liu, Susan B. 826, 1367, 1368, 1379 

Liu, Sze Hang Kevin 1550 

Liu, Tianhui 1404 

Liu, Ting 1952 

Liu, Wanqing 56


Liu, Wei 710, 719, 723 

Liu, Xiao 890, 991, 1015, 1630, 1644 

Liu, Xiaoying 658 

Liu, Xiu-Ping 366 

Liu, Xuan 1198 

Liu, Xue-Jing 935 

Liu, Yali 2061 

Liu, Yan 1198, 1595, 1675 

Liu, Yang 1678, 2021 

Liu, Yingxia 2048 

Liu, Yuhan 1611 

Liu, Yun 1383 

Liu, Yunlong 1332 

Liu, Zhang-Xu 2135 

Liu, Zhaohui 728 

Liu, Ziyi 1374 

Liu, Li 1939 

Livingston, Stephen 1798, 1807 

Livnat, Yuval 1032 

Lizarzábal, Maribel 595 

Llach, Josep 2084 

Lledo, Jose Luis 378 

Lleo, Ana 73, 602, 634 

Llop, Elba 1511 

Llorente-Izquierdo, Cristina 59 

Llovet, Josep M. 253, 254, 860 

Llovet, Patricia 1889 

Lloyd, Carla 1234 

Lo, Chung-Mau 70, 1746, 2000 

Lo, Gin-Ho 745 

Lo, Pei-Shuan 2040 

Lo, Sum Team 2225 

Lo, Tracie 1286 

Lo, Victor 1998 

Lo, Ying-Ru 577 

Lo Re, Vincent 1488 

Lo Turco, Edson G. 240 

Loaeza del Castillo, Aurora 595 

Lobach, Iryna 841 

Lobritto, Steven J. 1731 

Locarnini, Stephen 32, 1551, 1557, 

1563, 1566, 2022 

Locatelli, Luigi 1408 

Locati, Massimo 602 

Locklear, Cameron T. 575 

Loehr, Hanns F. 1170, 1200 

Loggi, Elisabetta 1888 

Lohse, Ansgar W. 76, 523, 1803 

Lok, Anna S. 67, 94, 576, 1555, 1850, 

1877, 2015, 2052 

Lombardero, Manuel 121 

Lomberk, Gwen 692 

Londjon-Domanec, Isabelle 1163, 1179 

Londoño, María-Carlota 982 

Long, Gang 992 

Long, Michelle T. 2191 

Long, Zhengyu 2035 

Long, Zhifeng 166 

Longato, Lisa 337, 1373, 1388, 1407, 

1445 

Longerich, Thomas 2128 

Longhi, Maria Serena 300, 608 

Longo, Diane M. 224, 228 

Loo, Nicole 743 

Loomba, Rohit 45, 236, 239, 913, 

1425, 1428, 1431, 1435, 1720, 

2149, 2150, 2153, 2154, 2201, 

2220, 2239 

Loomes, Kathleen M. 133, 1693, 1694 

Loomis, Timothy P. 93, 1786 

Lopategi, Aritz 2087 

Lopes, Marta 1033 

Lopez, Rocio 855, 856, 2162, 2182, 

2186 

Lopez Gomez, Marta 1511 

Lopez-Andujar, Rafae 1251 

López-Nevot, MAngel 583 

Lopez-Talavera, Juan-Carlos 1106 

Lopez-Vicario, Cristina 2087 

Lorber, Margalit 1032 

Lorent, Kristin 814 

Loss, George E. 1242 

Lotersztajn, Sophie 1288, 1391 

Lotteau, Vincent 1652 

Lotto, Martin 1752 

Loughran, Patricia 184 

Louie, Stan G. 1936 

Louie, Vincent 1135 

Loustaud-Ratti, Veronique 777, 1162, 

1187 

Louvet, Alexandre 145, 264, 1269, 

2082, 2222 

Louwers, Lisa 850, 1271 

Lovell, Sandra S. 250 

Loveridge, Robert 1775 

Low, Sarah E. 1928 

Lowe, Andrew 1100 

Lowe, Patrick 496, 1318, 1330 

Lowy, Elliott 540 

Loy, Veronica 1172 

Loyer, Xavier 887 

Lozada, Maria E. 127 

Lozano-Sepulveda, Sonia A. 1022 

Lu, Fengmin 34, 1634 

Lu, James T. 161 

Lu, Jie 692 

Lu, Junfeng 2061 

Lu, Mei 15, 525, 1578, 1789, 1800 

Lu, Minqiang 353, 375 

Lu, Quiajin 613 

Lu, Shelly C. 953, 1952, 2244 

Lu, Sheng-Nan 1593, 1604, 1828, 

1873, 2010 

Lu, Sizhao 60 

Lu, Wei 2023 

Lu, Wei-Yu 678 

Lu, Yuanfang 1910 

Lua, Ingrid A. 1354 

Luangmonkong, Theerut 1434 

Lubel, John 442, 483 

Lucena, M. I. 225, 583, 595, 596, 

1933 

Lucey, Michael R. 1231 

Lucidarme, Damien 1072 

Lucidi, Cristina 2067 

Ludwig, Andreas 100 

Luedde, Tom 828, 1924 

Lueth, Stefan 1629 

Luetjohann, Dieter 922, 961 

Luetkemeyer, Anne 40, 726 

Luft, Olga 1297 

Lugea, Aurelia 904 

Lui, Ka Yin 353, 375 

Luk, Kevin 1597 

Lukacs-Kornek, Veronika 43, 1299, 

1385 

Luke, Amy 532, 559 

Luketic, Velimir A. 52, 607, 609, 628, 

715, 1309, 1472, 2235 

Lukose, Thresiamma 272, 1454 

Luli, Saimir 44 

Lumeng, Lawrence 948 

Luminos, Luminita 2043 

Lunel-Fabiani, Françoise 1162 

Lunghi, Giovanna 1743, 1747 

Luntz, Steffen P. 1227 

Luo, Donghan 39 

Luo, Jian 106, 627, 1986 

Luo, Jianhua 670 

Luo, Mei 904 

Luo, Qingyang 1242 

Luo, Xiaoping 909, 1684 

Luo, Xun 1727, 1729 

Luo, Yan 1161, 1198 

Luo, Yuhuan 139, 142, 907 

Luo, Yunjun 159 

Luong, Tu Vinh 337, 413, 1373, 1388 

Lupinacci, Lisa Chiacchierini 707 

Lustig, Robert H. 1730 

Lutchman, Glen A. 1182, 1189 

Lutz, Karen 635, 645 

Lutz, Philipp 1008, 1287, 2077 

Lutz, Thomas 1060, 1081, 1156 

Luyendyk, James P. 81 

Luz, Rodrigo 273, 289 

Lv, Juan 1149 

Lv, Jun 1616 

Lv, Yi 698 

Ly, Mytop 1549 

Lynn, Amanda M. 643 

Lytvyak, Ellina 641 

M. Yoshida, Eric 308 

Ma, Ariel 549 

Ma, Hong 1660 

Ma, Hsiao-Yen 186 

Ma, Jimmy 2091 

Ma, Li 500 

Ma, Lina 2061 

Ma, Linyuan 2107 

Ma, Mang M. 204 

Ma, Mang-Ming 303 

Ma, Xiaoli 2025 

Ma, Xiong 308 

Ma, Yun 300, 305, 608 

Maan, Raoel 445, 1099, 1166 

Maasoumy, Benjamin 1131, 1572, 

1597, 1797 

Mabileau, Guillaume 1845 

Macaigne, Gilles 1072, 2082 

MacArthur, Meaghan 11 

MacConell, Leigh 239, 635, 644, 645 

Macdonald, Douglas 1904 

Macdonald, Graeme A. 1264 

Macdonald, Stewart 113 

MacDonald - Ottevanger, Sigrid 1846 

Macé, Vincent 264 

Macedo, Guilherme 1107, 1540 

Macera, Margherita 1837 

MacGregor, Louis W. 1830 

Machado, Joao 1256 

Machado, Mariana V. 897, 936, 960, 

1386, 2188 

Machicao, Victor I. 1513 

Machida, Keigo 682, 1918, 1936, 

1945, 2118 

Machineni, Surendra 2261 

Machnes, Ziv 730 

Macías-Rodríguez, Ricardo 828, 1471, 

1508


Mack, Cara L. 1692, 1699, 1706, 

1709 

Mackey, Aaron J. 175, 915, 1916 

Mackey, Rachel H. 2228 

Macklin, Jared 1350 

Mackman, Nigel 81 

MacLachlan, Ian 2007, 2062 

MacLachlan, Jennifer H. 1566, 2060 

MacNaughtan, Jane 908, 1300 

MacQuillan, Gerry C. 848, 2099 

Maddox, Avery 823 

Madejon, Antonio 1028 

Madireddi, Malavi 141 

Madrigal, Pedro 258 

Madsen, Bjørn S. 773, 780, 789, 792, 

1440 

Madsen, Emilie A. 1437 

Maeda, Haruka 1642 

Maeda, Kenji 1655 

Maekawa, Shinya 406, 1027, 1565 

Maes, Michaël 586 

Magaldi, Lora 1864, 1880 

Magdaleno, Fernando 1382, 2124 

Magee, John C. 133, 1694 

Maggart, Michael 1223 

Maggi, Mario 882, 902 

Magistroni, Paola 1 

Magni, Carlo F. 222 

Magni, Paolo 2187 

Magnusson, Baldur 2257 

Mahachai, Varocha 416 

Mahajan, Indra 1945 

Mahale, Parag 1030, 1160 

Mahalingam, Devalingam 1900 

Maharshi, Sudhir 1473 

Maher, Jacquelyn J. 306 

Mahgoub, Amar 1184, 2006 

Mahmoud, Huda 1480 

Mahmoud, Sahah A. 1011 

Mahoney, Douglas W. 169 

Mahtab, Mamun A. 2157 

Maier, Martin A. 36 

Maieron, Andreas 1092, 1885 

Maillard, Aline 132 

Maillette de Buy Wenniger, Lucas 637 

Maimone, Sergio 288, 1476 

Main, Janice 1826 

Maini, Mala K. 1626 

Maione, Sabatino 1837 

Maire, Laurence 1083 

Mairiang, Pisaln 416 

Maitland, Hillary 761 

Maitland-van der Zee, Anke H. 225 

Maiwall, Rakhi 83, 103, 146, 262, 

694, 1288, 2070, 2092 

Major, Marian E. 1546, 1854 

Makar, George A. 536 

Makino, Yuki 231, 453, 669, 1082, 

1357, 1661, 1985, 2130, 2136 

Makoto, Sanomura 966 

Makris, Alexia 1784 

Malago, Massimo 337, 1388, 1517 

Malato, Yann 98 

Malaya, Irina 721 

Maldonado, Eduardo N. 319 

Malek, Nisar P. 1183 

Malhi, Harmeet 58, 1315 

Malhotra, Pawan 1659 

Maliakkal, Benedict 82, 248, 250, 570, 

1478 

Malik, Mohammad U. 1255 

Malinverni, Raffaele 221, 1884 

Malinverno, Federica 1743, 1747, 

1751 

Malipatel, Renuka 284 

Mallano, Alessandra 1802, 1844 

Mallat, Ariane 421, 449, 1123, 1232 

Mallet, Maxime 744, 1510 

Mallet, Vincent 1194, 1803 

Mallolas, Josep 210 

Malta, Fernanda 48 

Maltez, Fernando 1859 

Man, Tak Yung 678 

Manch, Richard A. 1111, 1155, 1164 

Mancinelli, Romina 817 

Mandorfer, Mattias 149, 270, 475, 

561, 732, 755, 771, 1204, 1466 

Mandrekar, Pranoti 1351 

Maneschi, Elena 882, 902 

Manesis, Emanuel K. 2019 

Mangia, Alessandra 91, 249 

Mangini, Chiara 629 

Manini, Matteo A. 1747 

Manka, David 175, 664, 915, 1916 

Manka, Paul P. 1779, 1978, 2143 

Manley, Jr., Michael W. 650, 1767 

Mann, Derek 44, 652, 1381, 1408 

Mann, Jelena 1381 

Mannalithara, Ajitha 88, 1208, 2155 

Mannem, Arun 1753 

Mannino, Federica 1127 

Manns, Michael P. 76, 243, 444, 445, 

616, 712, 1002, 1045, 1049, 1084, 

1130, 1131, 1166, 1572, 1861, 

2026, 2128 

Mano, Yohei 476, 1643 

Manoharan, Muthiah 36 

Manolakopoulos, Spilios 2012, 2019 

Manon-Jensen, Tina 1437 

Manousou, Pinelopi 413, 801, 1445 

Manser, Christine N. 76 

Mantovani, Lorenzo G. 571, 1453 

Mantry, Parvez S. 704, 722, 1039, 

1084, 1146, 1169 

Manyakin, Nikolai 599 

Manzano Núñez, Fátima 691 

Mao, Qianguo 2048 

Mao, Shennen A. 2 

Mao, Tin 2258 

Marabita, Francesco 602 

Maragkos, Konstantinos 1126 

Maras, Jaswinder S. 581, 1005, 1344 

Marasco, Giovanni 741 

Maraver, Marta 1863 

Maraver Zamora, Marta 2171 

Marc, Abrams 510 

Marcellin, Patrick 241, 243, 887, 998, 

1557, 1601, 1847, 1860, 2014, 

2025, 2037, 2044, 2045, 2059 

Marchesini, Giulio 2164 

Marconi, Vincent 1488 

Marcus, James H. 1745 

Marcus, Sonja 380, 457 

Mare, Ruxandra G. 795 

Marek, George W. 138 

Marenco, Ted 730 

Marengo, Andrea 2209 

Margolis, Mary Kay 1467, 1485 

Margotti, Marzia 1888 

Marhenke, Silke 2128 

Marianelli, Leonardo 1752 

Maricic, Igor 1345 

Marier, J.F. 1915 

Marin, Jose J. 506 

Marin, Veronica 1339, 1346, 2098 

Marinho, Rui T. 150, 1859 

Mariño, Zoe 982 

Marins, Ed G. 1597 

Mariotti, Valeria 811 

Marisi, Giorgia 435 

Mariz, Daniela M. 2072, 2076 

Marker, Amanda E. 1694 

Markmann, James F. 837, 1732 

Marks, Kristen M. 1094 

Marks, Philippa 1083 

Marmon, Tonya 75, 155, 317, 628, 

635, 644, 645, 1458 

Maroni, Luca 21 

Marotta, Paul 1246 

Marquardt, Jens U. 677 

Marques, Carlos Eduardo C. 762 

Marques, Dalila M. 804 

Marquet, Pierre 1162 

Marquez, Max 11 

Marra, Fabio 973, 1410 

Marra, Fiona 1052, 1131, 1140 

Marrero, Idania 1345 

Marrero, Jorge A. 371 

Marrero Colon, Wesley J. 67 

Marri, Smitha 436 

Marroncini, Giada 949, 1963 

Marrone, Aldo 1837 

Marrone, Giusi 337 

Mars, Wendy M. 670, 1979 

Marschall, Hanns-Ulrich 76, 308, 662, 

810 

Marsh, Christopher L. 861, 872 

Marsh, Philip 47 

Marshall, Aileen 413, 489, 490, 1904 

Marshall, Vanessa 1154 

Marshall, Vincent D. 395 

Marshall, William L. 725, 730 

Martay, Kenneth 878 

Marti, Francesc 673 

Martí-Rodrigo, Alberto 1913 

Martin, Aaron 1906 

Martin, Daniel 774 

Martin, Jennifer H. 225 

Martin, Natasha K. 1794, 1830 

Martin, Paul 1054, 1128, 1148, 1206, 

2256 

Martin, Ross 91 

Martin, Steven R 1576 

Martin-llahi, Marta 378 

Martinello, Marianne 1083 

Martinet, Wim 894 

Martinez, Allyson 830 

Martinez, Anthony D. 1823 

Martinez, Elisa 717, 729 

Martinez, Jose Luis 1511 

Martinez, Mercedes 1731, 2129 

Martinez, Miguel 734 

Martinez, Olivia M. 341, 2123 

Martínez, Jorge A. 1217 

Martinez Rodriguez, Leonardo A. 2223 

Martinez Wassaf, Maribel 1752 

Martinez-Arranz, Ibon 953, 2163, 2244 

Martinez-Chantar, Maria L. 953, 2244 

Martinez-Llordella, Marc 1117 

Martinez-Lopez, Erika 1855


Martini, Silvia 1 

Martinot-Peignoux, Michelle 998, 1601, 

2044, 2045 

Martins, Americo 1256 

Martins, Eduardo B. 241, 2014, 2025 

Martins, Flaviano S. 2200 

Martins, Paulo N. 845 

Marukian, Svetlana 175, 1916 

Marusawa, Hiroyuki 1989 

Maruyama, Hitoshi 1490, 1516 

Maruyama, Kageshi 1433 

Maruyama, Toru 1912 

Marx, Steven 1042, 1050, 1068, 1069 

Marzioni, Marco 21, 76, 602, 621, 

1802, 1844 

Masaki, Keiichi 986, 1056, 1627, 

1649, 1681, 1895, 1944 

Masaki, Naohiko 794, 1138, 1658, 

1795, 2158, 2193 

Maslov, Alexander Y. 515 

Masola, Valentina 299 

Mason, Andrew 73, 76, 303, 628, 

634, 641 

Mason, Susan 1077 

Massaro, Joseph M. 2191 

Massarwa, Muhammad 1878 

Massella, Maurizio 1802, 1844 

Massetto, Benedetta 1076, 2004, 2015, 

2043, 2052, 2059 

Massey, Veronica L. 587, 1327, 2142 

Massie, Allan 1727, 1729 

Masson, Neil 753 

Masson, Steven 1216 

Massoud, Omar I. 1174, 1792 

Mast, T. Christopher 717, 729 

Masumoto, Akihide 1157 

Masuoka, Howard C. 320, 884, 948, 

1903 

Masur, Henry 92, 220 

Masur, Jack 578 

Mateo, Gloria 930 

Matern, Hugo 1986 

Mateus, Elia S. 1256 

Mathei, Catharina 1842 

Matheny, Michael E. 529 

Matherly, Scott C. 52, 1309, 1472 

Mathew, Arun 1261, 1276 

Mathias, Anita 1133, 1707, 2265 

Mathias, Wilson 1484, 1494, 1495 

Mathur, Karan 420 

Mathurin, Philippe 263, 1269, 2082, 

2222 

Matilla, Ana 378 

Mato, Jose M. 953, 1952, 2163, 2244 

Matono, Tomomitsu 1499 

Matsubara, Tsutomu 517, 592 

Matsubara, Yasuo 505 

Matsubayashi, Sunao 2097 

Matsuda, Hidetaka 944, 1682 

Matsuda, Shuya 1565 

Matsuda, Takashi 1447 

Matsuda, Yasunobu 2165 

Matsui, Takeshi 1097, 1655, 1778, 

1983 

Matsumoto, Akihiro 1673 

Matsumoto, Nobuyuki 1446 

Matsumoto, Shuichi 2097 

Matsumoto, Tomonori 1989 

Matsumoto, Toshihiko 329, 1447 

Matsumoto, Yoshihiro 2074 

Matsumoto, Yoshinari 191 

Matsunaga, Kazuhito 760, 1464, 1492 

Matsunaga, Kotaro 1446 

Matsunami, Kayoko 1006 

Matsushita, Hiroshi 1327 

Matsuura, Bunzo 1293 

Matsuura, Kentaro 1006 

Matsuura, Yoshiharu 1643 

Matsuzaki, Yasushi 970 

Matsuzawa, Naoto 1377 

Matta, Bassem 1887 

Matta, Benjamin M. 338 

Matta, Laura 309, 1153 

Matthews, Gail 210, 1083, 1826, 

1868 

Mattos, Angelo A. 469 

Matwiy, Trudy 1576 

Mauer, Amy S. 58 

Maul, Timothy M. 216 

Maurice, James B. 113, 764 

Maurice, Michèle 196 

Maurice, Sean B. 1413 

Mauss, Stefan 1060, 1078, 1081, 

1156, 1559, 1861 

Maverakis, Emanual 606 

Mawatari, Seiichi 799, 937, 1421, 

1838 

May, Sarah B. 357 

Mayatepek, Ertan 1227 

Mayayo-Peralta, Isabel 492 

Maybüchen, Lara 961 

Mayhew, Christopher 676 

Maynard-Muet, Marianne 1162 

Mayne, Tracy J. 75, 155, 313, 317, 

537, 546, 562, 1457, 1458 

Mayo, Marlyn J. 73, 106, 627, 634 

Mayo, Rebeca 620, 2244 

Mayo, Tara 36 

Mayumi, Iijima 1642 

Mazagova, Magdalena 59, 1319 

Mazariegos, George V. 763 

Mazer, Laura 1477 

Mazo, Daniel F. 1033, 2221 

Mazza, Giuseppe 337, 1388 

Mazzaferro, Vincenzo 1751 

Mazzarelli, Chiara 1751 

Mazzella, Giuseppe 609 

Mazzotta, Francesco 249 

McCaleb, Michael L. 183, 1353 

McCallister, Katharine 548 

McCann, Adam 565 

McCaughan, Geoff 106, 627, 746, 

1049, 1077, 1084, 1243, 1264 

McCauley, Bryan 607, 610 

McClain, Craig J. 114, 926, 1303, 

1304, 1305, 1306, 1312, 1331, 

1334, 2141 

McClintock, Kevin 2007 

McCone, Jonathan 715 

McConville, John F. 544 

McCormick, Aiden 1275, 2069 

McCormick, Joseph B. 1424 

McCorriston, Debra 871 

McCoy, Annette 1709 

McCullough, Arthur J. 239, 1265, 

2153, 2154, 2162, 2182, 2186, 

2220, 2239 

McCullough, Rebecca L. 334, 1308, 

2132 

McCune, Anne 564 

McDaniel, Kelly 834, 835, 929, 1317, 

1349 

McDonald, Stuart A. 101 

McGeough, Matthew D. 1371 

McGill, Mitchell R. 585, 586, 598, 

1767 

McGillicuddy, John W. 1272 

McGilvray, Ian 11 

Mcglone, Cindy 530 

McGuire, Brendan M. 1174, 1250, 

1768 

McHutchison, John G. 38, 91, 96, 205, 

219, 249, 616, 624, 632, 737, 739, 

746, 777, 1034, 1045, 1049, 1076, 

1080, 1094, 1105, 1120, 1128, 

1130, 1428, 1435, 1442, 2015, 

2037, 2052, 2149, 2239 

McIntyre, Chris 1480 

McKiernan, Patrick J. 2105 

McKiernan, Susan 1103 

McKillop, Iain H. 653 

McLauchlan, John 1173 

Mclaughlin, Leo 534 

McLaughlin, Mary 92, 220 

McLeman, Lindsay 1104 

McLeod, Marie-Ange 1588, 2053 

McLin, Valerie A. 1728, 1735 

McMahan, Rachel 938, 959, 1708 

McMahon, Brian J. 125, 626, 1790, 

1798, 1807 

McMahon, Peter S. 1052 

McMillin, Matthew 1507 

McMullen, Megan R. 334, 1308, 1346, 

2121, 2132 

McNally, John 205, 249, 2265 

McNiven, Mark A. 1310 

McNulty, Gwen L. 202 

McPhee, Fiona 702, 709 

McVicker, Benita L. 1969, 2139 

Medeiros, Roseane P. 1033 

Mederacke, Ingmar 1957 

Medici, Valentina 2109 

Medina, Vivian G. 1945 

Medina-Cáliz, I. 583, 595, 596 

Medmoun, Mourad 554 

Meehan, Kolin 493 

Mega, Ines 1256 

Megnien, Sophie 105, 162, 2145 

Mehal, Wajahat Z. 805, 979, 1209, 

1284, 2179 

Mehrab-Mohseni, Marjan 1023 

Mehta, Anand 371 

Mehta, Gautam 113 

Mehta, Neil 4, 50, 53, 414, 471 

Mehta, Nicita 496 

Mehta, Rajiv 241, 2014, 2025 

Mehta, Rajni 16 

Mehta, Rohini 891, 956 

Mehta, Shivang 1513 

Mei, Ruqi 1657 

Mei, Xiaonan 854 

Meillon, Jean-Christophe 2266 

Meissner, Eric G. 1029 

Mejia, Alejandro 1169 

Mejias, Caroline A. 548 

Mekaru, Steven 556 

Melancon, Sir Norman T. 1223 

Mele, Caterina 432 

Melgar, Jennifer 78 

Melgar-Lesmes, Pedro 1292


Melis, Marta 166 

Mello, Tommaso 902, 949, 1963 

Mellone, Massimiliano 1978 

Mells, George F. 155, 601, 1457 

Mells, Jamie 19 

Melton, Phillip 2185 

Meltzer, David O. 152 

Melum, Espen 258, 819, 833 

Memon, Muhammad S. 1872 

Menchise, Alexandra 1695 

Mendão, Luís 1859 

Mendes-Braz, Mariana 342 

Mendez-Bocanegra, Angela 2084 

Méndez-Sanchéz, Nahum 595 

Meng, Fanyin 190, 454, 514, 621, 

817, 834, 835, 929, 1317, 1349 

Meng, Li-Na 1465 

Meng, Xiaochun 1755 

Mengual, Edgardo 595 

Mennone, Albert 805, 829 

Menon, Krishna 1775 

Menon, Rajeev 1066, 1088 

Mensa, Federico J. 41, 248, 250, 705, 

719 

Mensing, Sven 1066 

Merchant, Michael L. 587 

Mercier-Darty, Melanie 1700 

Merion, Robert M. 838 

Méritet, Jean-François 30, 997 

Merlen, Grégory 179 

Merli, Manuela 2067 

Merniz, Mohamed 1621 

Meroueh, Fadi 1796 

Merriman, Raphael 538 

Merritt, Elliot 1117 

Merrouche, Wassil 450, 2181 

Mertens, Joachim C. 1402 

Mesarwi, Omar 1422 

Meshrekey, Raymond 1076 

Metcalfe, Leanne N. 532, 559 

Metivier, Sophie 219, 777 

Metselaar, Herold J. 68, 1238 

Metwally, Sherif N. 1280 

Meyer, Bernhard 444 

Meyer, Kirstin 617 

Meyer, Timothy 344, 1530, 1904 

Meyers, Rachel 36 

Mezzabotta, Lavinia 2209 

Mgaieth, Sara 483 

Mi, Yu-Qiang 1465 

Mi, Zhiyong 2143 

Miailhes, Patrick 1676, 1677 

Mian, Ajmal 798 

Miao, Chuan L. 552 

Miao, Ruoyu 1222 

Micco, Lorenzo 1626 

Michaels, Anthony 874, 1260, 1280 

Michalak, Thomas I. 1641 

Michalopoulos, George K. 670, 1770, 

1979 

Michel, Gerd L. 1812 

Michel, Maurice 1519 

Michel-Treil, Veronique 1597 

Michelotti, Gregory A. 66, 936, 962, 

1386, 2137 

Michielsen, Peter P. 797, 894, 976 

Michl, Patrick 1519 

Micolochick Steuer, Holly 2009 

Middleton, Michael S. 2150 

Midorikawa, Yutaka 407 

Mieli-Vergani, Giorgina 300, 608, 

1704, 1711 

Miethke, Alexander G. 823, 1695 

Miguez, Marcio Q. 2189 

Mikels-Vigdal, Amanda 1379 

Miki, Daiki 663, 986, 1645, 1649, 

1681, 1858, 1895, 1944 

Miki, Koichiro 2031 

Miki, Masahiro 1981 

Mikolajczyk, Adam E. 544 

Milana, Martina 203 

Milani, Stefano 1963 

Milián-Medina, Lara 1913 

Milkiewicz, Piotr 76 

Millán, Raquel 1863 

Millen, Rosemary M. 996, 2262 

Miller, Anne 529 

Miller, Charles M. 840, 1271 

Miller, Colton M. 507 

Miller, George 1282 

Miller, Gregory 784 

Miller, Lesley 1835 

Miller, Michael D. 91, 700, 703, 713, 

718, 1168, 2021 

Milligan, Scott 1046, 1108 

Millis, J. Michael 1229 

Mills, Anthony M. 716 

Miloh, Tamir A. 1716 

Milstein, Stuart 36 

Miltiadous, Oriana 253, 254 

Min, Albert 1095, 1165 

Min, Hae-Ki 905 

Minami, Masahito 886 

Minami, Tatsuya 461 

Minczeles, Noémie 1237 

Mine, Tetsuya 314, 383, 697 

Minello, Anne 1158, 1187 

Ming-jong, Bair 1996 

Mingarelli, Eleonora 21 

Minguez, Beatriz 378, 380, 438, 457 

Minichini, Carmine 431, 1543 

Minnis-Lyons, Sarah E. 678 

Minniti, Caterina 1498 

Minomi, Kenjiro 1375, 1378 

Minsart, Charlotte 178 

Minteer, William B. 4, 108 

Minter, Freneka F. 529 

Minuk, Gerald Y. 361, 1149 

Miquel, Juan F. 2210 

Mirabella, Stefano 1 

Miraglia del Giudice, Emanuele 1837 

Miranda, Joana P. 1921 

Mirolo, Massimiliano 602 

Mirshahi, Faridoddin 807, 905, 1309 

Mirza, Darius 1234 

Mirza, Roshni 2078 

Mirzac, Angela 730 

Mirzazadeh, Ali 1793 

Misaki, Ryo 1642 

Mishra, Alita 575 

Mishra, Bibhuti 130, 682 

Mishra, Lopa 130, 682, 2118 

Misra, Sandeep 170 

Missale, Gabriele 355 

Mistry, Nipun 682 

Mistry, Sameer 163 

Misu, Hirofumi 990, 1010 

Misurski, Derek A. 1089 

Mita, Eiji 1197, 1584, 2213 

Mitamura, Kuniko 804 

Mitchell, Angela M. 1708 

Mitchell, Ben C. 2021 

Mitchell, Donald G. 1906 

Mitchell, Joanne 1077 

Mitchell, Kristen A. 1403 

Mitchell, Natalie 1257 

Mitchell, Paul D. 136 

Mitchell, Tim 2099 

Mitov, Mihail I. 673 

Mitra, Lalita G. 103, 2070 

Mitsuhashi, Shuji 1394 

Mitsuya, Hiroaki 1655 

Mitsuyoshi, Hironori 886, 888, 901, 

1980, 2230 

Mittal, Rasham 404, 1171, 1569, 1619 

Mittal, Sahil 244, 357 

Mitterhofer, Anna Paola 983 

Miuma, Satoshi 952, 1671 

Miura, Mika 1027, 1565 

Miyaaki, Hisamitsu 952, 1671 

Miyabe, Katsuyuki 396 

Miyadera, Hiroko 1138 

Miyagawa, Koichiro 593, 883 

Miyagawa, Shinichi 869 

Miyagi, Takuya 1636 

Miyajima, Ichiro 304, 1819 

Miyake, Rei 569, 2140, 2234 

Miyake, Teruki 1293, 2146 

Miyake, Yasuhiro 339, 2236 

Miyaki, Eisuke 986, 1150, 1649, 1670, 

1681, 1944, 2120 

Miyamoto, Yukiko 1319 

Miyanishi, Koji 1984 

Miyata, Takashi 820 

Miyazaki, Masayuki 1157 

Miyazaki, Miyono 1375 

Miyazaki, Teruo 970 

Miyoshi, Eiji 906, 1642, 1809, 2156 

Miyoshi, Kenichi 1499 

Miyoshi, Masato 164 

Miyoshi, Takahiro 477 

Mizerski, Jacek 2043 

Mizokami, Masashi 91, 476, 794, 

1080, 1105, 1138, 1448, 1577, 

1614, 1632, 1643, 1658, 1673, 

1778, 1795, 2158, 2193 

Mizuguchi, Hiroyuki 1690 

Mizukoshi, Eishiro 370, 392, 430, 

1348, 1653 

Mizuochi, Tatsuki 676, 1724 

Mizutani, Kayo 906, 2156 

Mkada, Helmi 786 

Mo, Frankie 344 

Mo, Hongmei 91, 92, 219, 220, 713, 

718, 1080, 1105, 1120, 1168 

Mo, Lein-Ray 1996 

Moaven, Omeed 957, 1938 

Mobashery, Niloufar 708, 714 

Mochida, Hatsune 947 

Mochida, Satoshi 1059, 1064, 1474, 

1486, 1577, 1757, 2241 

Modaresi Esfeh, Jamak 1894 

Modi, Apurva A. 1146, 1199 

Modi, Deval 724 

Moeini, Agrin 253, 254 

Moeller, Frederick G. 1491 

Moestrup, Søren K. 2071 

Mogalian, Erik 2265 

Mogilenko, Denis 928 

Mogul, Douglas 1727, 1729


Mohamed, Essa A. 1966 

Mohamed, Sofiane 1009 

Mohammad, Mohammad K. 1303, 

1334 

Mohankumar, Swathi 358 

Mohanty, Sujata 690 

Mohr, Ashley M. 60, 1969 

Mohr, Rosemary 521 

Mokhtarani, Masoud 1467, 1478, 

1485 

Mole, Larry A. 93, 1786 

Molenkamp, Richard 208, 1846, 1995 

Moles, Anna 652 

Molina, Esther 2032 

Molleston, Jean P. 133, 159, 1694 

Molokanova, Olena 955, 1353 

Molokhia, Mariam 225 

Molrine, Deborah C. 704 

Momen-Heravi, Fatemah 109, 512 

Moncsek, Anja 1402 

Mondal, Kalyani 1986 

Monegal, Ana 615 

Monereo Muñoz, Maria Blanca 1341 

Monga, Satdarshan (Paul) S. 24, 255, 

821 

Monge, Fanny 891, 892, 956, 1428, 

2192 

Monico, Jesus 1178 

Monico, Sara 1743, 1747, 1751 

Monneret, Denis 786, 1510 

Monraats, Melanie A. 1238 

Monsour, Howard P. 1215 

Montagnese, Sara 292 

Montalbano, Marzia 1751 

Montalbano, Mauro 1991 

Montalva, Eva 1251 

Montane, Eva 596 

Montano-Loza, Aldo J. 303, 308, 616, 

641, 1481 

Monteiro, Eduardo d. 2248 

Montemezzo, Mauricio 2248 

Montestruc, François 1621 

Monti, Monica 1190 

Montialoux, Helene 3, 95 

Montin, Umberto 1751 

Monto, Alexander 1041 

Mony, Shruti 1155 

Mookerjee, Rajeshwar 908, 1300, 

1482, 1517 

Moon, Hyuk 504 

Moon, Scott 1041 

Mooney, Tim 36 

Moonka, Dilip 307, 1254 

Moore, Ann 1111, 1155, 1164 

Moore, Kevin 113, 337, 1388, 1407 

Moore, Raymond M. 1965 

Moorjaney, Divya 1456, 1461 

Moorman, Anne C. 15, 525, 1789, 

1799, 1800 

Moorman, Jonathan P. 984 

Mor, Eytan 875 

Moradpour, Darius 221, 1672, 1884 

Morales, Amilcar 1866 

Morales-Ibanez, Oriol 1327 

Morales-Ruiz, Manuel 65, 659 

Moran, Lauren 36 

Moran-Salvador, Eva 1381 

Morando, Filippo 148, 292 

Moratalla, Alba 1300 

Morbey, Ana 1256 

Moreau, Richard 83, 145, 178, 265, 

1288, 1391 

Moreira, Rebeca 266 

Morel, Philippe 2174 

Moreland, Anita L. 1154 

Morelli, Annamaria 882, 902 

Morelli, Giuseppe 1057 

Morelli, Maria Cristina 1751 

Moreno, Christophe 178, 1051, 1091, 

1179, 2079 

Moreno, Gigi 1042, 1068 

Moreno-Planas, Jose María 583, 2171 

Moreo, Kathleen 1141 

Morey, Tristan 89, 1090, 1112 

Morgan, Jake R. 152 

Morgan, Phillip 1098 

Morgan, Sarah 1455 

Morgan, Timothy R. 724, 1041, 1129 

Morgan-Stevenson, Vicki 946 

Mori, Amit 17 

Mori, Federica 958 

Mori, Kohjiroh 2156, 2202 

Mori, Trevor A. 2185 

Moriguchi, Michihisa 886, 901, 1980, 

2230 

Morihara, Daisuke 385, 410 

Morikawa, Hiroyasu 429, 1125, 1824 

Morikawa, Kenichi 287, 1302, 1672, 

1946 

Morillas, Rosa María 2032 

Morillo, Ramón 1863 

Morinaga, Maki 917 

Morio, Kei 472, 1893 

Morio, Reona 1893 

Morisco, Filomena 355 

Morishita, Naoki 1082, 1195, 1197, 

1201, 2213 

Morita, Chie 2031 

Moriuchi, Akihiro 799, 937, 1421, 

1838 

Moriwaki, Hisataka 657 

Moriya, Kyoji 461, 912 

Moriyasu, Fuminori 1047 

Morizono, Syusuke 1159 

Morley, Roland 1920 

Morling, Joanne R. 2168 

Moro, Tadashi 1369 

Moroianu, Serban A. 295 

Moroney, Justin B. 493 

Morosawa, Tatsuki 1026, 1648, 1943, 

2167 

Morris, Adam R. 1346 

Morris, Jason 522 

Morris, Jeffrey S. 130 

Morris, Meghan D. 1793 

Morrison, Dee 530 

Morrow, Bernice 943 

Morse, Gene D. 332 

Morton, Ryan 307, 420 

Mosaad, Youssef 1236 

Mosadeghi, Sasan 7 

Mosca, Nicola 431 

Moscat, Jorge 232 

Moscatelli, Alessandro 355 

Moser, Catherine D. 169, 396, 1965, 

1966 

Moser, Stephan 1885, 1886 

Moshage, Han 591, 612, 969, 1398, 

1416 

Mosoian, Arevik 1397 

Moss, Stephen E. 64 

Mossanen, Jana C. 1756 

Mössner, Belinda 792 

Motamed, David B. 1095, 1185 

Motola, Daniel L. 778 

Motomura, Kenta 1157, 1159 

Motoyama, Hiroaki 869 

Motoyama, Hiroyuki 1125 

Mott, Justin L. 60 

Mottram, Carl 1470 

Moucari, Rami 2044, 2045 

Mouillot, Thomas 264 

Mounzer, Karam 1136 

Mousa, Omar 1720 

Moussa, Adel 2266 

Moustafa, Hamdy M. 858 

Mouzaki, Athanasia 2094, 2095 

Mouzaki, Marialena 2184, 2238 

Moy, Carolyn 1864, 1880 

Moya, Angel 1251 

Moylan, Cynthia A. 161, 540, 889, 

1142 

Møller, Holger J. 1300, 2071 

Møller, Linda S. 780, 789, 1440 

Mridha, Auvro R. 55 

Mrzljak, Anna 1113, 1744 

Mtegha, Marumbo P. 1712 

Mu, Fan 711 

Mu, Xianmin 1910 

Muczynski, Kimberly 1221 

Mudan, Satvinder 599 

Mueller, Olaf 2188 

Mueller, Tobias 76 

Mugavero, Michael 1804 

Muir, Andrew J. 96, 106, 616, 624, 

627, 632, 1435, 2239 

Mukai, Kaori 1636 

Mukaida, Naofumi 2119 

Mukaide, Motokazu 1614 

Mukamal, Kenneth 2227, 2228 

Mukherjee, Amitava 197 

Mukherjee, Sunil K. 1659 

Mukhopadhya, Ashis 1104 

Mukhopadhyay, Chinmay 1344 

Mulazzani, Lorenzo 905 

Mulkay, Jean-Pierre 1842 

Mullen, Alan 778 

Mullen, Michael P. 1112 

Muller, Michael 1924 

Müller, Clara 43 

Müller, Niklas F. 308 

Müllhaupt, Beat 46, 221, 1402, 1818, 

1884 

Mulligan, David C. 9 

Mumtaz, Khalid 350, 874, 1260, 1280 

Mumtaz, Shaham 532, 559 

Mund, Ross 1899 

Mungiole, Nicole S. 521 

Munir, Samina 2166 

Munoz, Santiago J. 202 

Muñoz, Carolina 378 

Muñoz, Linda E. 1022, 1281 

Muñoz, Nina M. 2118 

Muñoz-Durango, Natalia 971 

Muñoz-Yagüe, Teresa 896, 939 

Munteanu, Mona 786 

Munzy, Donna M. 129 

Murad, Mohammad H. 282 

Muragundla, Anjaneyulu 118 

Murai, Kazuhiro 1082, 1661


Murai, Kazuhisa 990, 994, 1000, 1010 

Murakami, Eisuke 1858 

Murakami, Kazunari 2267 

Murakami, Masashi 970 

Murakami, Seishi 168, 990, 1010 

Murakami, Shuko 1599, 1655 

Murakami, Yoshiki 429, 1125, 1824 

Murakami, Yuya 1375 

Murakawa, Miyako 164, 406, 1018, 

1027, 1959 

Murali, Ramachandran 1936 

Muraoka, Masaru 1565 

Murata, Kazumoto 794, 1138, 1614, 

1658, 1673, 1778, 1795, 2158, 2193 

Murata, Yasuhiro 1640 

Muratori, Luigi 308 

Muratori, Paolo 308 

Murayama, Asako 1003 

Muromachi, Kaori 799, 937, 1838 

Muroyama, Ryosuke 505 

Murphy, Brian J. 141 

Murphy, Rinki 2176 

Murphy, Susan K. 889 

Murphy, Trudy 1546 

Murray, John W. 321, 333 

Murray, Karen F. 133, 1694, 2043 

Murthy, Karnam S. 807 

Musa, Hawah 1122 

Musolino, Cristina 1654 

Musto, Kaitlyn R. 1245 

Musukuma, Kalo 1575 

Mut, Deniz 1739 

Mutchnick, Milton G. 420 

Mutimer, David J. 1045, 1049, 1130, 

2003 

Mutsaers, Henricus A. 1427, 1434 

Mutsuki, Taiji 1157 

Mwinyi, Jessica 519, 520 

Myers, Robert P. 616, 624, 632, 737, 

739, 746, 777, 1428, 1435, 1442, 

2149, 2239 

Myronovych, Andriy 916 

Nabinger, Sarah C. 436 

Nadal, Elena 1270 

Nader, Fatema 18, 315, 316, 1460 

Nadig, Satish 1272 

Nadji, Mehrdad 914 

Nadler, Jerry L. 807 

Nafady, Mohammed 858 

Nafisi, Shirin 1111, 1155, 1164 

Naga Prasad, Sathyamangla V. 651, 

667, 668, 671, 1308 

Nagahara, Akihito 1851 

Nagai, Masato 551 

Nagamatsu, Hiroaki 399 

Naganuma, Atsushi 477 

Nagaoka, Katsuya 513, 1987 

Nagaoka, Shinya 408, 783, 1600 

Nagaoki, Yuko 1056 

Nagaoki, Yuuko 472, 1893 

Nagarkatti, Mitzi 962, 2133, 2137 

Nagarkatti, Prakash 962, 2133, 2137 

Nagasawa, Hideko 995 

Nagase, Syoji 1159 

Nagase, Toshihiko 1197, 2213 

Nagata, Hiroko 164, 406, 1018, 1027, 

1959 

Nagata, Naoto 144, 1348, 1968 

Naggie, Susanna 1035, 1094, 1142 

Nagy, Laura E. 675, 1308, 1339, 

1346, 2121, 2132 

Nahass, Ronald 40, 219 

Nahmias, Yaakov 331, 672, 679 

Nahon, Pierre 145, 362, 1808, 1847 

Naik, Hetanshi 2111, 2112 

Naik, Jahnavi 1095 

Naik, Sandhia 1571 

Naiki, Kayoko 1064 

Naing, Aung 1160 

Nair, Devaki 113 

Nair, Rashmi T. 1267, 2081 

Nair, Satheesh 1212, 1233 

Nairat, Samir S. 1843 

Naito, Masafumi 796 

Naito, Tatsushi 944, 1682, 2119 

Naito, Yoshiki 465 

Nakabori, Tasuku 97, 231, 669, 1082, 

1357, 1661, 1985, 2130, 2136 

Nakagawa, Ai 291 

Nakagawa, Hayato 461 

Nakagawa, Hidetoshi 370, 392 

Nakagawa, Hidewaki 1895 

Nakagawa, Kentarou 1584 

Nakagawa, Mina 164, 406, 1018, 

1027, 1959 

Nakagawa, Ryo 505 

Nakagawa, Shigeki 364, 2174 

Nakagome, Yu 1648, 1943, 2167 

Nakagomi, Ryo 461 

Nakahara, Takashi 986, 1056, 1649, 

1681, 1895, 1944 

Nakai, Masato 287, 1672, 1946 

Nakajima, Hisakazu 901 

Nakajima, Shunsuke 927 

Nakajima, Tomoaki 1116, 2065, 2207 

Nakakuki, Natsuko 1115 

Nakamoto, Nobuhiro 181, 569, 2131, 

2140, 2234 

Nakamoto, Shingo 1207, 1628 

Nakamoto, Yasunari 509, 944, 1682, 

2119 

Nakamura, Ikuo 1047, 1420 

Nakamura, Jun 2030 

Nakamura, Masato 1207, 1628 

Nakamura, Mikiko 1014, 1395, 1653 

Nakamura, Minoru 640 

Nakamura, Ryota 1522 

Nakamura, Shinichiro 2236 

Nakamura, Takuya 1648 

Nakamura, Toru 688, 1819 

Nakamura, Yoshiko 788, 1293 

Nakamura, Yuki 472, 1893 

Nakamuta, Makoto 707, 1157, 1159 

Nakanishi, Hiroyuki 1115 

Nakano, Chikage 466, 770, 781, 

1533, 1669, 1975 

Nakano, Masahito 455, 1819 

Nakano, Norihito 472, 1893 

Nakanuma, Yasuni 347, 614, 820, 

1901 

Nakao, Kazuhiko 952, 1671 

Nakao, Masamitsu 1577, 1757 

Nakao, Sachie 1369 

Nakashima, Hiroyuki 324 

Nakashima, Kazuhisa 456 

Nakashima, Masahiro 324, 952 

Nakashima, Osamu 465, 1990 

Nakashita, Shunya 296 

Nakatani, Kazuki 592 

Nakatsuka, Takuma 461 

Nakatsura, Tetsuya 509 

Nakayama, Nobuaki 1059, 1064, 

1474, 1486, 1577, 1757 

Nakayama, Yasuhiro 1565 

Nakazawa, Manabu 1474 

Nakazawa, Takahide 482 

Nakazuru, Shoichi 1584 

Naksuk, Niyada 1497 

Nalpas, Catherine 1179 

Nam, Byung Ho 377, 486 

Nam, Joon Yeul 474, 479, 1610, 2016 

Nan, Yuemin 638 

Nangia, Saachi 280, 560 

Nanni, Oriana 435 

Naoe, Hideaki 513, 1912, 1987 

Napoli, Andrew A. 217 

Naqvi, Alissa 568, 1093 

Naqvi, Syed J. 98 

Narciso-Schiavon, Janaína L. 2075 

Narimatsu, Hisashi 1448 

Naritaka, Nakayuki 1696 

Narkewicz, Michael R. 1706, 1708, 

1714 

Nart, Deniz 1739 

Nascimbeni, Fabio 2164 

Nascimento e Silva, Rafaella 518, 651, 

667, 668 

Nash, Michelle 1359 

Nasser, Imad 2160 

Nasti, Alessandro 325 

Natarajan, Sathish Kumar 60 

Nathan, Hari 395 

Nathanson, Michael H. 661 

Natsuizaka, Mitsuteru 1946 

Nattermann, Jacob 1008, 1287, 2077 

Naugler, Willscott E. 10, 69, 172, 481 

Naumann, Uwe 1170, 1200, 1861 

Navaratnam, Annalan M. 764 

Navarra, Giuseppe 1654 

Navarro, J. M. 1863 

Navarro, Victor J. 530, 535, 597, 

1922, 1923, 1932 

Navarro-Zornoza, Maria 8 

Navasa, Nicolás 953 

Nawa, Takatoshi 26 

Nawaz, Arif A. 1872 

Nawaz, Mohammad 1827 

Nayak, Suman 262 

Nayer, Ali 914 

Naylor, Paul H. 420 

Nazario, Hector E. 1146, 1169, 1199 

Nazef, Naim 2116 

Ndugga, Nambi J. 1451 

Ndungu, Milka 1146, 1199 

Neben, Steven 208, 2263 

Nederveen, Aart 2152 

Neff, Guy 1106 

Neff, Maria 1452 

Negrin Dastis, Sergio 1091 

Negro, Francesco 221, 1539, 1818, 

1884, 2174 

Negus, Susan 125, 1798, 1807 

Neijenhuis, Myrte 293 

Nejak-Bowen, Kari 24, 255, 821 

Nekachtali, Ouardia 772 

Nelson, David R. 94, 716, 989, 1057, 

1826 

Nelson, Kirbylee K. 1240 

Nelson, Mark 210


Nemoto, Tomoyuki 944, 1682 

Nephew, Lauren D. 865 

Nerenz, David R. 1792 

Nesher, Eviatar 875 

Neubauer, Stefan 2190 

Neuberger, James 631 

Neumann, Helmut 1506 

Neumann-Haefelin, Christoph 1666 

Neurath, Markus F. 1506 

Neuschwander-Tetri, Brent A. 157, 239, 

1535, 2150, 2153, 2154, 2220 

Nevens, Frederik 73, 171, 609, 634, 

1107 

Neves Souza, Lara 1733 

Nevzorova, Yulia A. 1924 

Newberry, Carolyn 536 

Newberry, Elizabeth P. 911, 1361, 

1365 

Newell, Evan W. 167 

Newell, Karen 1452 

Newstrom, David 632, 1359, 2149 

Newton, Jennifer M. 579 

Newton, Kimberly P. 159 

Ng, Anthony 2214 

Ng, Michel 1151, 1185 

Ng, Teresa 41, 248, 250, 705 

Ng, Vicky L. 133 

Ng, Vivian 488 

Ngo, Yen 786 

Nguyen, Anh-Hoa 2015 

Nguyen, Bach-Yen T. 40, 210, 251, 

700, 703, 715 

Nguyen, Huy 1549 

Nguyen, Justin H. 1236, 1245 

Nguyen, Khanh 1549 

Nguyen, Kristy 945 

Nguyen, Linda 1848 

Nguyen, Long H. 386, 458 

Nguyen, Michael D. 386, 458, 1816 

Nguyen, Mindie H. 123, 207, 359, 

367, 368, 386, 387, 388, 428, 458, 

459, 1182, 1189, 1561, 1668, 1805, 

1806, 1816, 1848, 2009, 2250 

Nguyen, My T. 1182, 1549 

Nguyen, Nghia H. 207, 359, 368, 

386, 388, 458, 1182, 1189, 1561, 

1806, 1816, 1848, 2250 

Nguyen, Ngoc Tue X. 685 

Nguyen, Pauline 387, 428, 1848 

Nguyen, Peter T. 386, 388, 458, 1816, 

1848 

Nguyen, Phirum 913 

Nguyen, Quy P. 514, 813 

Nguyen, Tuyen M. 36 

Nguyen, Vincent G. 207, 368, 1561 

Nguyen-Khac, Eric 206, 264, 1072, 

1187 

Ni, Hong-Min 140, 598, 1325 

Ni, Liyun 1133, 1707 

Ni, Yen-Hsuan 1701, 1705 

Nicholls, Hayley T. 881 

Nick, Elisabeth 2086 

Nickenig, Georg 961 

Nickkholgh, Arash 1227 

Nicolas, Carine 1009 

Nicolas, Gregory 394 

Nicoletti, Paola 225, 1930, 1933 

Nicoll, Amanda J. 311, 442, 483 

Nie, Yuanyang 904 

Niederau, Claus 1861 

Nielsen, Mette J. 119, 780, 1432, 

2178 

Niemietz, Christoph 2103 

Niemietz, Christoph J. 684 

Nierhoff, Dirk 687 

Nieto, Leonardo 1674 

Nieto, Natalia 1382, 1940, 2124 

Nietzsche, Sandor 808 

Nigar, Sofia 272 

Niinomi, Takuro 411 

Niitsu, Yoshiro 1375, 1378, 1433 

Nikolopoulou, Vasiliki 2094, 2095 

Nimanong, Supot 467 

Ning, Qin 909, 1656, 1660, 1683, 

1684 

Ning, Shunbin 984 

Ninomiya, Masashi 1026 

Nio, Kouki 699, 1953 

Nischalke, Hans Dieter 1008, 2077 

Nishida, Nao 1138, 1632, 2158 

Nishida, Naoshi 1935 

Nishiguchi, Shuhei 466, 770, 781, 

1533, 1669, 1975 

Nishijima, Norihiro 1989 

Nishikawa, Hiroki 466, 770, 781, 

1533, 1669 

Nishikawa, Taichiro 886, 888, 901, 

1980 

Nishimura, Miyuki 1375, 1378 

Nishimura, Norihisa 689 

Nishimura, Takashi 466, 770, 781, 

1533, 1669, 1975 

Nishimura, Tatsuro 760, 1464, 1492 

Nishina, Sohji 995, 1937 

Nishino, Ken 2030, 2199 

Nishio, Akira 26, 1635, 1636, 1809, 

1982 

Nishio, Kumiko 1584 

Nishio, Takahiro 402, 1213 

Nishiyama, Kiyoshi 324 

Nishizaki, Yasuhiro 2177 

Nissen, Nicholas N. 1479 

Nitta, Sayuri 164, 406, 985, 1018, 

1959 

Nitta, Takeo 1901 

Nittono, Hiroshi 804, 1696 

Niu, Congrong 35, 2009 

Niu, Jing 754 

Niu, Junqi 1149, 1193, 1580, 1603, 

1620, 1657, 1869, 2018, 2058, 2107 

Niu, Xiaoxia 1536, 2008 

Nix, Sarah 335, 358 

Niyibizi, Nyiramugisha 1835 

Njouom, Richard 1833 

Njoya, Merlin 1597 

Nkuize, Marcel 1091 

Noel, Benoit 974 

Noel, Gillian 1737 

Nogata, Yoichi 918 

Noguchi, Kazunori 304 

Nogueira, Monize A. 240 

Noh, Choong Kyun 379, 1567, 2056 

Nojima, Masanori 1448, 1983 

Nojiri, Shunsuke 1448, 1954 

Nokes, Brandon 1459 

Noma, Naruto 1918 

Nomoto, Minoru 2165 

Nomura, Hideyuki 1097, 2031 

Nomura, Norihiro 1941 

Nomura, Yoshimoto 699, 1953 

Nong, Yunhong 1562 

Noon, Luke A. 691, 1364 

Noor, Bashir 2205 

Nordstrom, Beth L. 1175 

Norero, Blanca M. 1217 

Noronha Ferreira, Carlos 150 

Noronha-Dutra, Alberto A. 2237 

Norris, Suzanne 1103 

Northup, Patrick 761, 1258, 1501 

Notake, Tsuyoshi 869 

Notvall-Elkey, Lena M. 2022 

Noureddin, Mazen 2244 

Nousbaum, Jean-Baptiste 554, 2082 

Nouso, Kazuhiro 2236 

Novack, Victor 2224 

Noviello, Stephanie 706, 709, 711, 

716 

Nozaki, Yasutoshi 97, 231, 669, 1082, 

1357, 1661, 1985, 2130, 2136 

Nozaki, Yuichi 2207 

Nuamah, Rosamond 2105 

Nugent, FW 412 

Numthavaj, Pawin 1999 

Nunez, Marina 438 

Nuñez, Susana 899, 1320 

Nunn, Abigail D. 958 

Nusse, Roel 683 

Nwankwo, Chizoba 717, 727, 729 

Nyberg, Anders H. 87, 1788, 1825 

Nyberg, Lisa M. 87, 1788, 1825 

Nyberg, Scott L. 2 

Nydam, Trevor L. 838 

Ó Broin, Pilib 515 

O’Beirne, James 413, 489, 490, 764, 

793, 1455, 1530, 1904 

O’Bleness Gray, Nicole 1260 

O’Brien, April 830 

O’Brien, Christopher 2256 

O’Brien, Daniel R. 129, 1965 

O’Brien, Sean 396 

O’Donoghue, Pam 489, 490, 1904 

O’Grady, John G. 1744 

O’Hara, Steven P. 1994 

O’Leary, Jacqueline G. 82, 200, 570, 

1084 

O’Neil, Maura 352 

Oakes, Kath 646, 1098, 1113, 1588, 

2053, 2054 

Oakley, Fiona 44, 652, 1381, 1408 

Oaks, Zachary 1961 

Oberti, Frédéric 145, 263, 2188 

Obry-Roguet, Véronique 1093 

Ochi, Hidenori 986, 1056, 1645, 

1649, 1681, 1858, 1895, 1944 

Ochi, Noriyo 1573 

Ocho, Makoto 1448 

Oda, Kohei 799, 937, 1421, 1838 

Oda, Masaya 1514 

Odamaki, Toshitaka 2074 

Odena, Gemma 110, 1327 

Odin, Joseph A. 597, 607, 1095, 

1185, 1932 

Oe, Bibiana 316 

Oe, Shinji 593, 883 

Oehrle, Melissa 1695 

Oettl, Karl 2068 

Ofengeim, Dimitry 832 

Ofosu, Andrew 1749 

Ofotokun, Igho 1444 

Ofuji, Kazuya 509, 944


Ogasawara, Sachiko 1990 

Ogata, Kei 304, 1819 

Ogawa, Hirohisa 1981 

Ogawa, Koji 287, 1946 

Ogawa, Shintaro 1573, 1599 

Ogiso, Satoshi 1976 

Ogunwobi, Olorunseun 1967 

Ogura, Satoshi 227, 1974 

Ogwo, Cheri 1233 

Oh, Jae K. 1470 

Oh, Sohee 474, 1610 

Oh, Yumin 115 

Ohashi, Jun 1632 

Ohira, Hiromasa 551 

Ohira, Tetsuya 551 

Ohki, Takamasa 382, 2240 

Ohkusa, Toshifumi 2074 

Ohmae, Saori 1918 

Ohmoto, Masaki 524 

Ohmura, Takumi 1116 

Ohne, Kumiko 1573, 1599, 1618 

Ohno-Machado, Lucila 112 

Ohtake, Takaaki 927 

Ohtani, Masahiro 944, 1682 

Ohtsuka, Masato 1369 

Ohtsuru, Akira 551 

Oishi, Naoki 168, 699, 1348, 1653, 

1953, 1970 

Oja-Tebbe, Nancy 1578 

Ojiro, Keisuke 1001 

Oka, Takahito 2030 

Okabe, Haruka 314 

Okabe, Hirohisa 821 

Okada, Hikari 144, 1010, 1014, 1348, 

1377, 1395, 1653, 1953, 1968, 1970 

Okada, Toshihiro 188, 1420 

Okajima, Akira 886, 888, 2230 

Okajima, Hideaki 1213 

Okamato, Yuta 1642 

Okamoto, Koji 952 

Okamoto, Tomohiro 1420 

Okamoto, Toru 1643 

Okamoto, Toshiaki 1499 

Okano, Jun-ichi 1499 

Okanoue, Takeshi 707, 888, 2199 

Okazaki, Kazuichi 613 

Okimoto, Gordon S. 1553 

Okolicsanyi, Stefano 571 

Okoronkwo, Nneoma O. 295, 1181 

Okubo, Tomomi 291 

Okuda, Yukihiro 1213 

Okuno, Masayuki 402 

Okuse, Chiaki 1446 

Okushin, Kazuya 912 

Olcoz, Jose Luis 2171 

Olde Damink, Steven 604 

Olinga, Peter 1427, 1434 

Oliva, Giovanni 1476 

Olivares, Shantel 257 

Oliveira, Carlos 1900 

Oliveira, Claudia P. 240, 1033, 2164, 

2221 

Oliveira, Ketti G. 1855 

Oliveira, Mariana M. 1523 

Oliveira, Nuno G. 1921 

Oliveira, Yanaihara P. 2237 

Oliver, Gavin R. 1965 

Olivera-Martinez, Marco A. 748, 769 

Ollivier-Hourmand, Isabelle 145, 264, 

2082 

Oloruntoba, Omobonike O. 540 

Olsen, Mark 256 

Olson, Janet E. 397 

Olson, Jody C. 1291, 1814, 1815 

Olthoff, Kim M. 838, 853, 868 

Olyaee, Mojtaba S. 1814, 1815 

Olynyk, John K. 2185 

Omagari, Katsumi 1655 

OMahony, Megan 1172 

Omar, Rabab F. 1076 

Omata, Masao 1080, 1105 

Omori, Risa 1302, 1941 

Omura, Hitoshi 1000 

On, Sissi 887 

Onali, Simona 309, 413, 489, 490, 

1153 

Oniki, Kentaro 1912 

Onishi, Hideki 2236 

Onishi, Hiroka 799, 937, 1838 

Onishi, Yoshiki 26, 453, 1635, 1636, 

1809, 1982 

Onisto, Maurizio 299 

Onji, Morikazu 524 

Ono, Atsushi 986, 1649, 1681, 1895, 

1944 

Ono, Masafumi 2156, 2207 

Ono, Suzane K. 1544 

Onodera, Mio 1528 

Onorato, Lorenzo 431 

Onori, Paolo 621, 817, 834, 835 

Oo, Ye H. 1298, 1710, 2003 

Ooka, Kohtaro 1370 

Oosterhuis, Dorenda 1427, 1434 

Or, Yat Sun 2257, 2259, 2260 

Oribe, Junya 2267 

Oriishi, Tetsuharu 233 

Orimo, Tatsuya 866 

Orkin, Chloe 210 

Orlent, Hans 1091 

Orlicky, David J. 139, 142, 907, 921 

Orloff, Susan L. 69, 481 

Orman, Eric S. 1451, 1903 

Oró, Denise 659 

Orr, Anne 670, 1979 

Orr, David W. 2176, 2225 

Orr, James G. 2226 

Ortega, A. 583, 595, 1863 

Ortiz, Inmaculada 378 

Ortiz, Juan 78 

Ortiz, Pablo 2163 

Ortiz-Lasanta, Grisell 1128 

Oruç, Nevin 492 

Ory, Daniel S. 911 

Osafo-Addo, Awo 1731 

Osalde-Solís, Geraldine 1471 

Osawa, Yosuke 476 

Osgood, Gregory 200 

Oshige, Akihiko 799, 937, 1421, 1838 

Oshita, Masahide 2213 

Osinusi, Anu 92, 205, 220, 249, 713, 

1013, 2265 

Osna, Natalia A. 589, 1323, 1347 

Osorio, Robert W. 4, 538, 2104 

Ostrenko, Oleksandr 617 

Ota, Tsuguhito 495 

Otal, Philippe 263 

Otani, Satoshi 164, 406 

Otazua, Pedro 583 

Otero, P. A. 493 

Otgonsuren, Munkhzul 575, 892, 2173, 

2192, 2204 

Otsubo, Takeshi 2160 

Otsuka, Taiga 296 

Ott, Peter 768 

Otto, Benjamin 523 

Ottobrelli, Antonio 1 

Ottosen, Soren 2255 

Otvos, James 2227 

Ou, Xiaojuan 374 

Oude Elferink, Ronald 827 

Ouled Cheikh, Ibtissem 1621 

Oules, Valerie 1052 

Outhay, Malena 1223 

Ouwerkerk-Mahadevan, Sivi 39 

Ouyang, Elsa 1165 

Ouyang, Xinshou 805, 979, 1284, 

2179 

Ouzan, Denis 1052, 1847 

Overcash, J. Scott 41, 248, 250 

Overton, Edgar T. 1444, 1804 

Owens, Christopher M. 2257 

Owens, Jennifer 812, 822, 1363 

Owens III, Albert P. 81 

Owens-Grillo, Janet 644 

Oyakawa, Leticia 1855 

Ozaki, Yoshihiko 944, 1682 

Ozasa, Kotaro 551 

Ozaslan, Ersan 308 

Ozdogan, Osman C. 558, 1822 

Oze, Tsugiko 1195, 1197, 1201, 2213 

Ozeki, Itaru 1116, 1203, 1778, 2065 

Ozenne, Violaine 145 

Ozutemiz, Omer 246, 1247 

Pabst, Oliver 100 

Pacher, Pal 1374 

Pack, Michael 195, 814 

Padberg, Kornelius 183 

Padula, William V. 152 

Pagan, Sarah 611 

Page, Agata 1381 

Page, Kimberly 1793 

Pageaux, Georges-Philippe 3, 95, 1158 

Pai, Rish K. 107 

Paik, Seung Woon 422, 423, 485, 

1591, 1960, 2049 

Paik, Yong-Han 422, 485, 1415, 1591, 

1960, 1972, 2049 

Pais, Raluca 1239, 2164 

Pak, Judy 260, 753 

Pak, Stephen C. 193, 197 

Palaios, Emmanouil 850, 1271 

Palaniyappan, Naaventhan 1524 

Palazzo, Adam 2009 

Palazzo, Donatella 222, 983 

Palle, Sirish 12 

Palma, Elena 599 

Palmer, Daniel 344 

Pamecha, Viniyendra 695, 1268, 1340 

Pan, Calvin Q. 209, 1165, 1587, 

2015, 2052 

Pan, Chen 2023 

Pan, Jason J. 108 

Pan, Jen-Jung 1424 

Pan, Terry 1216 

Pan, Yang 919, 934 

Pan, Yu 1149, 1193, 1869, 2107 

Pan, Zhaoxing 1709, 1715, 1737 

Pan-ngum, Wirichada 555 

Panchal, Ankur 1397


Pandak, William M. 804 

Pande, Gaurav 274 

Pandey, Akshay 1988 

Pandit, Vanshja 1340, 2088 

Pandol, Stephen J. 904 

Panduro, Arturo 1855 

Pandya, Prashant K. 1814, 1815 

Pandya, Purva 510, 2116 

Panebianco, Deborah L. 730, 731 

Pang, Phillip S. 91, 96, 219, 247, 

1045, 1049, 1105, 1130, 1442, 1860 

Panigrahi, Rajesh 501, 506, 1096 

Pant, Kishor 1664 

Pantea, Victor 31 

Panzitt, Katrin 662 

Paoli, Pier P. 1381, 1408 

Papa, Salvatore 1978 

Papadaki, Sotiria 1489 

Papatheodoridis, George V. 241, 1126, 

2012, 2014, 2019 

Papiamonis, Nikolaos 2094 

Papp, Maria 2090 

Pappas, Stephen Chris 278 

Pappo, Orit 1878 

Paradis, Valerie 417, 480, 887 

Paraná, Raymundo 595 

Paranaguá-Vezozzo, Denise 1544 

Paranjpe, Shirish 670 

Pareja, María J. 2171 

Parekh, Krishan 1298 

Parés, Albert 73, 76, 308, 603, 615, 

620, 629, 634, 642 

Pariente, Alexandre 1072, 1072, 2082 

Parikh, Neehar D. 67, 395, 418, 1850 

Parisé, Hélène 1087, 1143 

Parisi, Ioanna 793, 1530 

Park, Beom Jin 433, 1438 

Park, Brandi 1835 

Park, Do Youn 1025 

Park, Haesuk 1079, 1538 

Park, Hana 1542 

Park, Hyeongmin 491 

Park, In-Whee 2056 

Park, James 1165 

Park, Jang-June 1001 

Park, Jin-Kyu 1322 

Park, Jong Sung 115 

Park, Joohan 1567 

Park, Joong-Won 377, 486, 851 

Park, Jun Yong 346, 1542, 1558, 1581 

Park, Jung Gil 363, 403, 749 

Park, Kyoung-Sook 950 

Park, Kyu-Sang 1401 

Park, Nomi 464 

Park, Ogyi 115 

Park, Rae Woong 2056 

Park, Seol-Hee 1358 

Park, Seong W. 658 

Park, So Hyun 807 

Park, Soo Young 363, 403, 426, 749, 

2028, 2038 

Park, Soohyun 1415, 1972 

Park, Su Cheol 685 

Park, Su-Hyung 1025 

Park, SuBum 876 

Park, Sun Seob 377 

Park, Sun Young 1567 

Park, Tae Jun 187 

Park, Young Nyun 1902 

Parker, Richard 574, 1321 

Parkes, Julie 776, 793, 1455, 1530, 

1856 

Parlak, Erkan 846 

Parrella, Korin 1151, 1870 

Parruti, Giustino 1107 

Parsons, Judith 1023 

Parvin-Nejad, Fatemeh P. 1364 

Pas, Suzan D. 1590, 2122 

Pascasio, Juan Manuel 2032 

Pascual, Sonia 378 

Pasetto, Maria Cristina 309, 1153 

Pasic, Fuad I. 1188 

Pasquale, Giuseppe 1543 

Pasquazzi, Caterina 222 

Pasquet, Blandine 145 

Passmann, Sandra 1559 

Passos-Castilho, Ana Maria 48 

Pastor, Helena 2171 

Pastor, Jose J. 194 

Patch, David W. 413, 764, 793, 1530 

Patel, Anna 1151 

Patel, Arpan A. 1512 

Patel, Dilip 260 

Patel, Forum 606 

Patel, Janki R. 913 

Patel, Keyur 205, 624, 1428, 1887, 

2169 

Patel, Neal M. 1095, 1185, 1202 

Patel, Rahul 1151 

Patel, Rajiv 1787 

Patel, Ruchi 1720 

Patel, Sameer 1775 

Patel, Samir 2147, 2208 

Patel, Trushar 1267, 2081 

Patel, Tushar 335, 358, 384 

Patel, Vishal C. 202, 740 

Patel, Mamta 1470 

Paternostro, Rafael 149, 1466 

Pathil, Anita 1411, 2195 

Patidar, Kavish 2235 

Patil, Mallikarjun 284, 2101 

Patra, Sharda 1663 

Pattanasirigool, Chaowalit 1631, 1662 

Patten, Daniel A. 1296 

Pattou, Francois 2222 

Paugam-Burtz, Catherine 1781 

Paul, Andreas 1779 

Paul, Jean-Louis 887 

Paul, Sonali 1592 

Paule, Bernard 421, 449 

Paulino, Lismeiry 545 

Pauta, Montserrat 65, 659 

Pauwels, Arnaud 264 

Pavel, Mihai-Calin 860 

Pavel, Oana 734 

Pavendranathan, Gokulan 1243 

Pavkov, Douglas 1172 

Pavletic, Steven 2144 

Pavlides, Michael 2190 

Pavlovic, Julie 1225 

Pavlovic, Vedran 245, 2026 

Pavone, Nicole 536 

Pavuk, Marian 2233 

Pawlik, Timothy M. 356 

Pawlinski, Rafal 81 

Pawlotsky, Jean-Michel 693, 987, 999, 

1123, 1700, 1797 

Pawlowska, Malgorzata 2043 

Payance, Audrey 480, 772, 2148 

Payer, Berit A. 1204 

Payerl, Doris 2068 

Payungporn, Sunchai 2063 

Péan, Noémie 816 

Pearlman, Brian 715 

Peck-Radosavljevic, Markus 37, 149, 

252, 270, 475, 561, 732, 755, 771, 

1049, 1058, 1092, 1204, 1466, 1525 

Pecriaux, Caroline 1194 

Peddu, Praveen 212 

Pedersen, Mark 1043, 1109, 1145, 

1224, 1241, 1262 

Pediconi, Natalia 958 

Pedley, Alison 2191 

Pedro, Ana Júlia 150 

Pedrosa, Marcos C. 16, 1106 

Peeples, Miranda 711 

Peeraphatdit, Thoetchai 397, 1497 

Pei, Luying 2007 

Peiffer, Kai-Henrik 980, 1559 

Peiser, Leanne 2009, 2052 

Pelaquier, Agnes 2082 

Pelemis, Mijomir 2004 

Pellegatta, Gaia 355 

Pelletier, Gilles 1237 

Pelletier, Shawn 1258 

Pellicoro, Antonella 62 

Peltekian, Kevork M. 204 

Peña, Jose M. 1881 

Penaranda, Guillaume 1009 

Pencek, Richard 155, 625, 628, 635, 

644, 645, 1458 

Pendergast, Jason 573 

Penders, John 100 

Pene, Veronique 30, 997 

Peng, Cheng-Yuan 40, 42, 1828, 1996, 

2040 

Peng, Haoran 353, 375 

Peng, Hong 1583 

Peng, Jie 1998 

Peng, Jin 828, 1924 

Peng, Kesong 22 

Peng, Lee 1122 

Peng, Lee F. 1015 

Peng, Liang 1423, 1772 

Peng, Lijun 131 

Peng, Shan S. 2107 

Peng, Yan-Zhong 2023 

Peng, Zhechu 1945 

Peng, Zhen-Wei 826, 1367, 1368, 

1379 

Penn, Michael 1177 

Penna, Giuseppe 893 

Pennell, Craig E. 2185 

Pepe, Veronica 1270 

Peppa, Dimitra 1626 

Perales, Celia 1028 

Peralta, Carmen 8, 326, 342 

Perarnau, Jean-Marc 145 

Perazzo, Hugo 786, 2189 

Perdigoto, Rui 1256 

Pereira, Cristiano A. 2200 

Pereira, Gustavo 84, 762, 2072, 2076 

Pereira, Isabel V. 586 

Pereira, João Luiz 84, 762 

Pereira, Marcus 272 

Pereira, Stephen P. 76 

Pereira, Thiago A. 936 

Perez, Renata D. 762 

Perez, Renata M. 84, 273, 289, 2189 

Perez-Cormenzana, Miriam 620


Perez-Hernandez, Onan 1341, 1342 

Perez-Rodriguez, Edelmiro 1281 

Perichon, Regis 161 

Perilla, Mauricio 1271 

Perinelli, Paola 222, 983 

Perini, Lisa 629 

Peris, Pilar 615 

Perito, Emily R. 1730, 1736 

Perkins, Neil D. 652 

Perkins, Susan 436 

Perl, Andras 1961 

Perlmutter, David H. 193, 197 

Pernazza, Alejandra 2106 

Perner, Dany 980, 1559 

Perno, Carlo Federico 203, 222 

Péron, Jean-Marie 263, 1803, 2082 

Perrone, Leonardo 1089 

Persico, Marcello 1802, 1844 

Perumalswami, Ponni V. 545, 1095, 

1151, 1185, 1202, 1783 

Perumpail, Ryan B. 199, 201, 541, 

877, 1534, 1745, 1750 

Pesci, David 1451 

Pessoa, Mario G. 1033 

Peter, Joy A. 1826 

Peter, Senaka 1867, 1883 

Peters, Marion G. 1094, 1444, 1570 

Peters, Yvette 644 

Petersen, Dennis R. 921 

Petersen, Joerg 37, 241, 252, 1058, 

1078, 1176, 1205, 1559, 2003, 2014 

Peterson, Ryan M. 32, 2022 

Petit, Laetitia Marie 1735 

Petoumenos, Kathy 1083 

Petraccia, Luisa 1190 

Petraksa, Supanna 1999 

Petrasek, Jan 180 

Petropoulou, Fotini 1126 

Petrov-Sanchez, Ventzislava 362, 1808, 

1847 

Petrtyl, Jaromir 1487 

Petta, Salvatore 973, 2164 

Peyret, Thomas 1915 

Peyton, Adam 1041, 2256 

Peyton, Diane 536 

Pflanz, Stefan 2009 

Pfreundschuh, Michael 463 

Phakdeekitcharoen, Bunyong 1999 

Pham, Huong T. 1186 

Phan, Jennifer 488 

Phaosawasdi, Kamthorn 416, 555 

Phelan, Kieran 676 

Phelps, Janet R. 2007 

Philip, Philip A. 420 

Philips, Cyriac A. 103, 2070 

Phillips, John D. 2111, 2112 

Phillips, Sandra 163 

Philosophe, Benjamin 1214 

Pho, Mai T. 152 

Phung, Van 1986 

Pi, Liya 989 

Pianko, Stephen 91, 249 

Piano, Salvatore 148, 266, 292, 767 

Piao, Shenglong 113 

Piard-Ruster, Karine 341 

Piazza, Nicholas 1257 

Picard, Nicolas 1162 

Picat, Marie-Quitterie 132 

Picchio, Gaston 39 

Piconese, Silvia 958 

Pieper, Dietmar H. 1002 

Pierce, Ruth 17 

Pieri, Giulia 1751 

Pilette, Christophe 1072 

Pillai, Anjana 369 

Pillardy, Jaroslaw 1841 

Pilot-Matias, Tami 705, 714, 1065, 

1084, 1086 

Piluso, Alessia 1190 

Pilutti, Chiara 148, 266, 292 

Pimentel, Carolina F. 1484, 1494, 

1495, 2160, 2227 

Pine, Karen 1452 

Pineton de Chambrun, Marc 1288 

Pinho, João Renato R. 48, 1033, 1855 

Pinkston, Christina 2233 

Pinna, Antonio Daniele 1751 

Pinotti, Rachel 2170 

Pinzani, Massimo 337, 413, 517, 764, 

1373, 1388, 1407, 1410, 1445, 1517 

Pio, Jose R. 87 

Piotrowski, Joy I. 1121 

Piovesan, Sara 775 

Piratvisuth, Teerha 1631, 1662 

Pirillo, Martina 1888 

Pirmohamed, Munir 225 

Pirola, Carlos J. 2216, 2217 

Pisano, Giuseppina 393 

Pisano, Maia Belen 1752 

Pisaturo, Mariantonietta 1543 

Piscaglia, Fabio 355, 602 

Pischke, Sven 523, 1803 

Pitrone, Alessia 288 

Pizarro, Margarita 969 

Plankey, Michael 1444 

Plat, Jogchum 922, 961 

Platt, Blake 71, 843 

Platt, Heather L. 40, 210, 380, 438, 

729 

Plompen, Elisabeth P. 1590 

Plotnik, Julia N. 626, 1790, 1798, 

1807 

Plumeier, Iris 1002 

Poca, Maria 734 

Pocha, Christine 16, 1884 

Pockros, Paul J. 94, 108, 251, 609, 

1039, 1057 

Poddar, Minakshi 255 

Podesser, Bruno K. 1525 

Podevin, Philippe 30 

Podlaha, Ondrej 1651, 1668, 2052 

Podsadecki, Thomas 1039, 1086, 1161 

Poerzgen, Peter 1121 

Pogemiller, Hope 572 

Pohl, Sabine 1519 

Pohnert, Georg 808 

Poiteau, Lila 999, 1833 

Poizot-Martin, Isabelle 568, 1093 

Pokora-Pachowicz, Agnieszka 1582 

Pokorski, Izabella 1343, 1450 

Pol, Stanislas 1194, 1808, 1847, 1860 

Polak, Wojciech G. 68 

Polepally, Akshanth R. 1051, 1066 

Pollard, Katherine A. 2121 

Pollicino, Teresa 1127, 1654, 1898 

Polo, Miriam 1913 

Polotsky, Vsevolod 1422 

Poluektova, Larisa Y. 589 

Polvani, Simone 949, 1963 

Polychronidis, Georgios 1227 

Pomfret, Elizabeth A. 71, 838, 843, 

847 

Pomper, Martin 115 

Pompili, Maurizio 432 

Pons, Fernando 378, 1511 

Ponsioen, Cyriel Y. 73, 74, 76, 604, 

623, 634 

Ponzoni, Mirco 44 

Poole, Lauren G. 587, 2142 

Poon, Art 1832 

Poon, Kok Siong 1812 

Poongkuran, Mugilan 371 

Poonia, Bhawna 1029 

Poordad, Fred 39, 41, 247, 248, 250, 

706, 716, 1051, 1065, 1086, 1433, 

2257, 2260 

Poovorawan, Kittiyod 416, 555, 1631 

Poovorawan, Yong 2036, 2063 

Pop, Oltin T. 1763 

Pope, Amanda 1716 

Popov, Yury 826, 1367, 1368, 1379 

Porat-Shliom, Natalie 20 

Porayko, Michael K. 279 

Porcella, Rita 309 

Porcherot, Marine 1652 

Pornthisarn, Bubpha 1631, 1662 

Porsche, Cara 938, 959 

Port, Kerstin 1131 

Portal, Andrew J. 564 

Porte, Robert J. 68, 1258 

Porter, John R. 195, 814 

Posa, Alessandro 432 

Pose, Elisa 266 

Posner, Amanda J. 1736 

Posthouwer, Dirk 1613 

Posuwan, Nawarat 2063 

Potenza, Nicoletta 431 

Poterucha, John J. 836, 1263 

Potosky, Darryn R. 1101, 1215 

Potze, Wilma 2249 

Poulsen, Kyle L. 1339, 1346 

Poupon, Raoul 73, 634, 642 

Pouriki, Sophia 1489 

Pourmand, Kamron 1826 

Pouteau, Michele 2044, 2045 

Povero, Davide 57, 969, 1371, 1971 

Powell, Elizabeth E. 784, 1362 

Poynard, Thierry 786, 1510 

Pradat, Pierre 568, 1093, 1162, 2003 

Prado, Veronica 1307 

Prall, Stacy 552 

Pratschke, Johann 276, 1222 

Pratte, Katherine 174 

Precoma, Dalton B. 2248 

Preisinger, Christian 1924 

Premkumar, Madhumita 1344, 2078 

Premoli, Caterina 1747 

Premont, Richard T. 66, 936, 1386 

Preston, Ta-Wanda 1864, 1880 

Pretorius, Carel J. 784 

Price, Angie 92 

Price, Jackie 2168 

Price, Patricia 2185 

Prieto, Martin 583, 596, 1230, 1251, 

2032, 2171 

Prins, Maria 1846 

Prip-Buus, Carina 887 

Pritilata, Rout 284 

Protopopas, George 1181 

Protzer, Ulrike 1684


Prough, Russell A. 666, 2233 

Provenzale, Dawn T. 540 

Provenzano, Angela 1410 

Puente, Angela 1511 

Puerto, Marta 1295, 1520 

Pugasab, Alongkorn 1999 

Pugatch, David 710 

Pugliese, Pascal 568, 1093 

Pullicino, Richard 2251 

Püngel, Tobias 1756 

Pungpapong, Surakit 1038, 1124, 

1236 

Punpanich, Dollapas 2089 

Puntes, Víctor 659 

Puoti, Massimo 1802, 1844 

Purcell, Robert H. 1646 

Puri, Puneet 52, 240, 905, 1309, 1316, 

1340, 1472, 2163, 2235 

Purnak, Tugrul 308 

Pursell, Natalie W. 510, 2116 

Pustavoitau, Aliaksei 1255 

Puszyk, William M. 1967 

Putignano, Antonella 1769, 1780, 1781 

Pyenson, Bruce 1069 

Pyrsopoulos, Nikolaos 295, 1181, 1827 

Qamar, Amir A. 412 

Qaqish, Roula B. 708, 714 

Qi, Xin 651 

Qi, Xingshun 754 

Qi, Xun 1998, 2002 

QI, Lingxia 2107 

Qin, Shengying 225 

Qin, Xian-Yang 657 

Qiu, Chunhui 353 

Qiu, Xinjie 919 

Qu, Jun 332 

Qu, Xiaowang 1001 

Qu, Xiumei 2123 

Quaglia, Alberto 1733, 1978 

Qualls, Charles 1916 

Quan, David J. 1560 

Quaranta, Maria Giovanna 1153, 

1802, 1844 

Quer, Josep 1021, 1674 

Quertinmont, Eric 178 

Questel, Franck 1102 

Quinn, Gemma 39 

Quinn, Matthew A. 1507 

Quintana, Albert 1295 

Quintero-Platt, Geraldine del Carmen 

1341, 1342 

Quintini, Cristiano 840, 1271 

Quispe, Wilber 515 

Qureshi, Huma 1872 

Qureshi, Kamran 1122 

Rabatin, Abigail 1142 

Rachakonda, Vikrant 1496, 1531 

Racila, Andrei 575 

Radaeva, Svetlana 1332 

Radenne, Sylvie 3, 95, 1158 

Radford, Peggy 1716 

Radhakrishnan, Kavita 1560 

Radreau, Pauline 1652 

Radu, Claudia M. 351 

Raffa, Giuseppina 1654, 1898 

Raffetti, Elena 1568 

Raftopoulos, Spiro C. 1264 

Raghunathan, Sahana 330 

Rahimi, Robert S. 1210 

Rahman, Adeeb 988, 1289 

Rai, Nitish 259 

Rai, Rahul 2245 

Raimondo, Giovanni 288, 1127, 1476, 

1654, 1802, 1844, 1898 

Rainteau, Dominique 816 

Rajagopalan, Ramakrishnan 1693 

Rajanayagam, Jeremy K. 1710, 1728 

Rajesh, S. 1340 

Rajwanshi, vivek 993 

Rakela, Jorge 211, 1761 

Rakhmanaliev, Elian 1812 

Rallier, Sandrine 1833 

Ramachandran, Priya 867, 1698 

Ramakrishna, Gayatri 327, 695, 1005 

Ramalho, Fernando 150 

Ramanathan, Meera 1145, 1241, 1262 

Ramani, Komal 594 

Rametta, Raffaela 2187 

Ramirez, Gilberto 1574 

Ramirez-Vega, Ruben 1199 

Ramji, Alnoor 722 

Ramos, Hilario 35 

Rana, Abbas 369 

Rana, Khurram 716, 726 

Ranabhat, Ganesh 80 

Ranchal, Isidora 1300 

Rao, Anuradha 19 

Rao, Huiying 782, 1436, 1820, 1850, 

1877 

Rao, Kiran V. 1181 

Rao, Swati 1122 

Rao, Vinaya 1233 

Raouf, Ahmed A. 1853 

Raoul, Jean-Luc 394 

Rapaccini, Gian Ludovico 355, 432 

Raseni, Alan 2098 

Rashid, Asif 130, 682, 2118 

Rasineni, Karuna 1310 

Raskopf, Esther 80 

Rastellini, Cristiana 1991 

Rastogi, Archana 327, 695, 1483, 

1762 

Ratziu, Vlad 105, 162, 737, 739, 786, 

1428, 1435, 2145, 2164, 2239 

Rau, Monika 2195 

Rauch, Andri 1575, 1818 

Raun, Loren 1991 

Rautou, Pierre-Emmanuel 81, 145, 265, 

772, 887, 1288, 2148 

Ravendhran, Natarajan 715 

Ravenelle, Francois 1333 

Ravindran, Karthik 420 

Rawal, Bhupendra 1245 

Rawat, Dinesh 1697, 1725 

Rawls, John F. 2188 

Ray, Kristen 1831 

Rayar, Michel 487 

Raybon, Joseph J. 728 

Raychaudhuri, Pradip 1988 

Rayhill, Stephen C. 1211 

Raymond, Daniel 1839 

Raymond, Eric 417 

Raymond, Valérie-Ann 179 

Raza, Roshan 136 

Razavi, Homie 1818, 1842, 1859, 

1872 

Razavi-Shearer, Devin M. 1872 

Razumilava, Nataliya 1994 

Re, Viviana 1752 

Reau, Nancy 205, 271, 522, 539, 544, 

553, 1065, 1813 

Rechling, Christian 1137 

Reddy, K. Gautham 271, 539, 544, 

553 

Reddy, K. Rajender 82, 217, 570, 712, 

743, 1034, 1039, 1045, 1049, 1057, 

1130, 1134, 1442, 1759, 1768 

Reddy, Mettu S. 859, 867 

Reddy, Sheela S. 1452 

Reding, Raymond 1734 

Redman, Rebecca 708, 714 

Reebye, Vikash 118 

Reesink, Hendrik W. 208, 251, 1633, 

1995, 2005, 2255, 2263 

Reeves, Helen 425 

Reggiani, Paolo 1747 

Regula, Jaroslaw 609 

Rehermann, Barbara 1650 

Rehman, Saad 1223 

Reiberger, Thomas 149, 270, 475, 561, 

732, 755, 771, 1204, 1466, 1525 

Reich, David J. 202 

Reichman, Trevor W. 1242 

Reid, Danielle 2143 

Reig, Ana 629 

Reig, Anna 603, 620 

Reig, Maria Elisa 378 

Reiling, Janske 818, 975 

Reiller, Brigitte 1796 

Reilly, Briget 536 

Reimundo, Pilar 1674 

Rein, David B. 156, 1792, 1839 

Reinoso, Humberto 318 

Reisch, Thomas 705 

Reisinger, Florian 2128 

Reissing, Johanna 828 

Rela, Mohamed 859, 867, 1698 

Ren, Hong 1044, 1647, 2048 

Ren, Jinma 774 

Ren, Junping 984 

Ren, Suping 33, 2064 

Ren, Wanhua 516 

Ren, Xiaowei 1316 

Rendon, Paloma 378 

Renelus, Benjamin D. 790 

Rengasamy, Asokan 1692, 1699 

Renner, Eberhard L. 11, 204, 542 

Rennison, Julie H. 518, 651, 667 

Renou, Christophe 1072 

Rentero Alfonso, Carles 64 

Renz, John F. 1229 

Requena, Silvia 1180 

Resche-Rigon, Matthieu 998, 1601 

Rescigno, Maria 893 

Reuben, Adrian 1914 

Reul, Winfried 961 

Reverter Segura, Enric 982 

Revill, Peter A. 1551, 1557, 1563 

Rewisha, Eman A. 1862, 1879 

Rey, David 1093 

Rey, Jorge 2106 

Reyes, Ella F. 71, 843, 847 

Reyes, Jorge 878, 1211 

Reynaert, Hendrik 1091 

Reynolds, Catherine 530 

Reynolds, Justin A. 1111, 1155, 1164 

Rezvani, Milad 98 

Rhame, Frank S. 1034 

Rhee, David B. 515


Rhodes, Kimberly 1099, 1857 

Riachi, Ghassan 264 

Riad, Joseph 1076 

Ribas, Vicent 899, 1320 

Ribeiro, Roberto M. 2221 

Ribera, Jordi 65, 659 

Riccardi, Laura 432 

Rice, Kenner 1374 

Rice, Thomas M. 1793 

Richards, Lisa M. 913, 1431 

Richards, Tara D. 216 

Richardson, Crit T. 279 

Richardson, Jason R. 919, 934 

Richardson, Peter 90, 156, 244 

Richen, Xavier 1796 

Richou, Carine 1036, 1187 

Riedl, Florian 1525 

Rieger, Armin 561, 1204 

Rieger, Megan 1945 

Riese, Peggy 1762 

Rietdijk, Svend 208 

Rife, Kelsey 1154 

Rigamonti, Cristina 1743 

Riggio, Oliviero 2067 

Rigou, Geraldine 974 

Rijckborst, Vincent 1585 

Rijnders, Bart J. 1007 

Rikner, Leif 810 

Rimassa, Lorenza 602 

Rimland, David 438, 1488 

Rimola, Jordi 860 

Rinaldo, Piero 2 

Rinaudo, Jo Ann 371 

Rincón, Diego 1503 

Rinella, Mary E. 1215, 1226, 1235 

Ringers, Jan 68, 1238 

Rinker, Franziska 2026 

Rinninella, Emanuele 432 

Riordan, Stephen 106, 627 

Rios, Miguel 734 

Ríos-Torres, Silvia L. 1471 

Ripoll, Cristina 1503, 1519, 1520 

Riquelme, Arnoldo 2210, 2231 

Rispoli, Rossella 1978 

Ritzenthaler, Jeffrey D. 2142 

Rius, Bibiana 2087 

Riva, Alessandra 1751 

Riva, Antonio 599, 2143 

Rivas-Estilla, Ana Maria G. 1022 

Riveiro-Barciela, Mar 1021, 1674 

Rivera, Elenita M. 218 

Rivino, Laura 167 

Rizvi, Bisharah S. 2144 

Rizwan, Maira 2100, 2114 

Rizza, Giorgia 1 

Rizzetto, Mario 1, 1802, 1844, 2209 

Ro, Weonsang S. 504 

Roa, Jhoanna 1094 

Roach, Jonathan 1692, 1699 

Roayaie, Sasan 851 

Robaeys, Geert 1613, 1842 

Robbins, Kristen N. 1715 

Robertazzi, Suzanne 1101 

Roberts, Benjamin R. 1291 

Roberts, Dean W. 1914 

Roberts, Jackson L. 1407 

Roberts, John P. 4, 50, 53, 72, 414, 

1219, 1273, 1560 

Roberts, Lewis R. 121, 127, 129, 169, 

381, 396, 397, 415, 851, 1124, 

1497, 1965, 1966 

Roberts, Mark S. 104, 151 

Roberts, Sheree 1298 

Roberts, Stuart K. 106, 249, 442, 483, 

627, 1077, 2262 

Roberts, Teri 1826 

Robertson, Avril A. 55 

Robertson, Michael 40, 42, 210, 251, 

700, 701, 703, 707, 712, 715, 717 

Robic, Marie Angele 145 

Robinson, Benjamin 337, 1388 

Robinson, Joseph L. 1479 

Robles, Camila 2210 

Robles Diaz, Mercedes 583, 595, 596 

Robson, Richard A. 1128 

Robson, Simon C. 608, 826, 1222, 

1394, 2227, 2228 

Rocha, Chiara 1742 

Rocha, Clarissa S. 63 

Roche, Bruno 1237 

Rockey, Don C. 737, 739, 1478, 1922, 

1931 

Rockstroh, Jürgen K. 37, 210, 252, 700, 

703, 1058, 1081, 1156 

Rodell, Tim 2015, 2052 

Roder, Heinrich 1900 

Roder, Joanna 1900 

Rodes, Juan 342 

Rodgers, Joel B. 1804 

Rodrigo-Torres, Daniel 1327 

Rodrigues, Cecília M. 176, 832, 897, 

940, 960 

Rodrigues, Livia 240, 2221 

Rodrigues, Pedro M. 176, 832, 897, 

940, 960 

Rodrigues, Teresa 150 

Rodriguez, Carla V. 1811, 1871 

Rodriguez, Christophe 1700 

Rodriguez, Eduardo A. 1206 

Rodríguez, Manuel 378, 2032 

Rodriguez de Miguel, Cristina 2084 

Rodriguez-Agudo, Daniel 804 

Rodriguez-Barradas, Maria C. 1488 

Rodriguez-Canales, Jaime 166 

Rodríguez-Córdova, Paola A. 1471 

Rodriguez-Frias, Francisco 1021, 1674 

Rodriguez-Garcia, Jose Luis 1508 

Rodriguez-Lope, Carlos 378 

Rodriguez-Torres, Maribel 718 

Rodt, Thomas 444 

Roeker, Lindsey 1263 

Rogal, Shari S. 1753 

Rogers, Jason 1151 

Roggendorf, Michael 31, 1021 

Rogler, Charles E. 515 

Rohatgi, Sarika 1718 

Rohel, Alexandra 3 

Rohr, Cristian O. 2217 

Rohrbach, Jeff 536 

Rojas, Angela 1863 

Rojas de Santiago, Pamela 969, 971 

Rojas Luengas, Vanessa 1012, 1297 

Rokosh, S. Rae 1282 

Rollin, Francois 1835 

Romagnoli, Renato 1, 1740 

Romain, Mélissa 887 

Roman, Eva 583, 1509 

Roman, Jesse 2142 

Roman, Sonia 1855 

Romano, Antonietta 148, 266, 292, 

775 

Rombouts, Krista 337, 517, 908, 1373, 

1388, 1407, 1410, 1445, 1517 

Romero, Miriam 1028 

Romero-Gomez, Manuel 105, 162, 

1134, 1863, 2145, 2164, 2171 

Romero-Marrero, Carlos J. 1894 

Rondon, Juan 2257 

Rong, Guanghua 354 

Ronot, Maxime 417 

Rooge, Sheetalnath B. 690, 694, 1483, 

1659 

Roos, Ruud A. 1667 

Roosblad, Jimmy 1846 

Rorive, Sandrine 178 

Rosa, Isabelle 132, 206, 567, 1072, 

2082 

Rosa, Jose Luis 1418 

Rosa, Julia S. 2075 

Rosales-Zabal, Jose Miguel 1863 

Rosato, Stefano 1802, 1844 

Rose, Christopher F. 51, 1481, 1520, 

1523 

Rosen, Charles B. 836 

Rosen, Hugo R. 28, 503, 938, 959, 

1430, 1706, 1708, 2085 

Rosenau, Jens 200 

Rosenberg, Arielle R. 30, 997 

Rosenberg, Gillian 1557, 1563 

Rosenberg, William M. 249, 776, 793, 

1455, 1530, 1856, 1904 

Rosenthal, Philip 133, 1694, 1730, 

1736 

Rosi, Silvia 148, 292 

Rosmorduc, Olivier 132, 417 

Ross, David 128, 540 

Rossaro, Lorenzo 1041 

Rosselli, Matteo 113 

Rossi, Francesca 1837 

Rossi, Giorgio 1743, 1751 

Rossi, John 118 

Rossi, Ric 2099 

Rossi, Simona 530, 535 

Rossi, Stephen 106, 198, 627, 1986 

Rossignol, Emilie 3 

Rosso, Chiara 2209 

Rosso, Natalia 1339, 1346, 2098 

Rota, Matteo 571, 1453 

Rota, Monica 1453 

Roth, Christopher G. 1906 

Roth, David 700, 703, 1054, 1148 

Roth, Nitzan 1350 

Rothenberg, Marc E. 887 

Rothstein, Kenneth D. 202 

Rothweiler, Sonja 826 

Rotllan, Noemi 65 

Rotman, Yaron 218, 2224 

Rouach, Hélène 30 

Roudot, Alice 105, 162, 974, 2145 

Roudot-Thoraval, Françoise 362, 421, 

449, 567, 1232, 1808, 1847 

Rougemont, Anne-Laure 1735 

Rougier, Hayette 1676, 1677 

Rouhani, Farshid 138 

Roura-Poch, Pere 2084 

Rousseau, Annick 1162 

Rousseau, Benoit 693 

Rouveau, Nicolas 1833


Roux, Olivier 265, 772, 1781, 2148 

Rowe, Ian A. 574, 1321 

Rowell, Richard 1478 

Roychowdhury, Sanjoy 2132 

Roytman, Marina 1121 

Roytrakul, Sittiruk 1999 

Rødgaard-Hansen, Sidsel 2071 

Ruane, Peter J. 205, 248, 716, 1065, 

1076 

Rubbia-Brandt, Laura 2174 

Rubenstein, Kevin 1811, 1871 

Rubin, Angel 1230, 1251 

Rucci, Paola 983 

Rude, Eric J. 1801 

Rude, Mary K. 2091 

Rudler, Marika 145, 744, 1510 

Rudnick, Sean R. 1501 

Rugge, Massimo 1889 

Ruhaak, L. Renee 606 

Rui, Zhang 1812 

Ruiz, Alex 595 

Ruiz, Alicia 1674 

Ruiz, Isaac 1123, 1269 

Ruiz de Galarreta, Marina 1382 

Ruiz-Cabello, Francisco 583 

Ruiz-Gaspa, Silvia 615 

Ruiz-Margáin, Astrid 828, 1471, 1508 

Rule, Jody A. 1761, 1776 

Runge, Jurgen H. 2152 

Runkana, Ashok 668 

Ruocco, Giancarlo 165 

Ruping, Kathleen 1181 

Rupp, Christian 618 

Rupp, Daniel 992 

Rupp, Loralee B. 15, 525, 1578, 1789, 

1799, 1800 

Ruscica, Massimiliano 2187 

Rusie, Erica 1141 

Russ, Kirk B. 1220, 1253 

Russell, Stephen J. 2 

Russell, Stuart D. 1255 

Russo, Aniello 431 

Russo, Francesco P. 1270, 1802, 1844, 

1889 

Russo, Mark W. 17, 597, 1023 

Rustgi, Vinod K. 1478, 1741 

Ryan, Karen K. 916 

Ryan, Marno C. 442, 483 

Ryan, Michael 219 

Ryan, Robert 1163, 1179 

Rychlicki, Chiara 21 

Ryoo, Minjung 594 

Ryska, Miroslav 721 

Ryu, Ho Sang 391, 433, 1504 

Sa-Cunha, Antonio 487 

Saab, Sammy 201, 1079, 1087, 1139, 

1143, 1144, 1152, 1462, 1512, 

1538, 1745 

Saad, Jennifer 1329, 2073 

Saad Hashem, Mohammed 1163 

Saag, Michael 210, 1804 

Saba, Francesca 2209 

Saberi, Behnam 1350 

Sabino, Ester C. 2221 

Saborowski, Anna 2128 

Sacco, Rodolfo 355 

Saccomanno, Stefania 21 

Sacerdoti, David 767 

Sachdeva, Sanjeev 1473 

Sack, Ulrich 261 

Sada, Yvonne 357 

Sadagoparamanujam, V.M. 2112 

Sadana, Prabodh 1338 

Sadikova, Ekaterina 1788 

Sadiq, Javaid 1234 

Sadiq, Omar 420 

Sadowsky, H. Steven 1235 

Sadozai, Hassan 1012, 1297 

Saeb-Parsy, Kourosh 258 

Saeed, Ali 612 

Saeki, Akira 408, 783, 1600 

Sætrom, Pål 118 

Safadi, Rifaat 660, 1532, 1878 

Safaeinili, Niloufar 563, 849 

Safer, Ricky 630 

Safran, Michal 1685 

Safwan, Mohamed 867, 1698 

Sagnelli, Caterina 1543 

Sagnelli, Evangelista 431, 1543, 1837 

Saha, Banishree 109, 496, 1016, 

1318, 2127 

Sahin, Hacer 963 

Sahney, Amrish 103, 146, 147, 262, 

2070, 2092 

Sahota, Amandeep K. 1119, 1135, 

1171, 1569, 1619 

Sahraoui, Salah 1621 

Sahu, Smiti S. 436 

Saibara, Toshiji 2156 

Saich, Andrew 537 

Saigal, Sanjiv 2245 

Saikia, Paramananda 2121, 2132 

Saini, Rajnish 260, 753 

Saito, Hidetsugu 181, 569, 2131, 

2140, 2234 

Saito, Keigo 509 

Saito, Masaki 1975 

Saito, Yoshinobu 97, 231, 453, 669, 

1082, 1357, 1661, 1985, 2130, 2136 

Saitoh, Satoshi 1062, 1191, 2020, 

2241 

Saitoh, Shinji 135 

Saitta, Carlo 288, 1476, 1654, 1898 

Saji, Yukiko 1201 

Sajwani, Karim 1128, 2265 

Sakae, Haruka 799, 937, 1838 

Sakai, Akira 551 

Sakai, Hiroshi 869 

Sakai, Takao 1390 

Sakai, Yoshio 325, 947, 1348, 1395, 

1653, 1968 

Sakai, Yoshiyuki 466, 770, 781, 1533, 

1669 

Sakaida, Isao 329, 760, 1447, 1464, 

1492 

Sakakibara, Yuko 1584 

Sakamori, Ryotaro 26, 97, 231, 453, 

669, 1082, 1197, 1357, 1635, 1636, 

1661, 1809, 1982, 1985, 2130, 2136 

Sakamoto, Minoru 1565, 1673 

Sakamoto, Naoya 287, 1632, 1672, 

1946 

Sakane, Sadatsugu 97, 1985 

Sakaue, Takahiko 688 

Sakisaka, Shotaro 385, 410 

Sako, Katsumi 456 

Sakuma, Tetsushi 1627 

Sala, Margarita 378 

Salama, Khaled M. 2215 

Salamat, Amjad 1872 

Salas, Eduardo 35 

Salazar-Gonzalez, Rosa-Maria 916 

Salehitezangi, Nazanin 563, 849 

Salerno, Debora 165 

Salgia, Reena 9 

Salhab, Ahmad 660, 1532 

Saliba, Faouzi 145, 1237, 1269 

Salizzoni, Mauro 1 

Sällberg, Matti 992 

Salloum, Shadi 25, 1015 

Salmeron, Javier 2171 

Salmon, Isabelle 178 

Salotti, Joanie 913 

Salustro, Claudia 309 

Salvadori, Ricardo 1495 

Salvatore, Lucia 432 

Salyana, Muhammad Atif 1282 

Salzer, Hannah 618 

Samaan, Maggie H. 840 

Samadi Kochaksaraei, Golasa 1576 

Samala, Niharika 2144 

Sambrotta, Melissa 1721 

Samet, Jeffrey 1488 

Samir, Waleed 1076 

Samonakis, Dimitrios N. 2094 

Samp, Jennifer C. 1089 

Sampaziotis, Fotios 258 

Samsonov, Mikhail 721 

Samstein, Benjamin 369, 838, 1454 

Samuel, Didier 421, 449, 487, 1045, 

1049, 1084, 1130, 1237, 1269, 

1782, 2082 

Samyn, Marianne 1711 

San Juan, Fernando 1251 

Sanabria, Judith 595 

Sanai, Faisal M. 1179, 1607 

Sanaka, Sirish 1122 

Sanchez, Ana M. 652 

Sanchez, Concepcion 1281 

Sanchez, Jorge 2166 

Sanchez, William 50, 53 

Sanchez, Yuri 1042, 1068, 1069, 

1087, 1143, 1144, 1462 

Sánchez, Araceli G. 1028 

Sanchez Ciceron, Adriana 595 

Sánchez-Pacheco, Aurora 1028 

Sanchez-Perez, Maria Jose 1341, 1342 

Sancho-Bru, Pau 1327, 2148 

Sander, Beate 350 

Sandlers, Yana 330 

Sandoval, Daniela 2210 

Sandoval, Darleen A. 916 

Sandrasegaran, Kumar E. 2150 

Sanford, Ukina 78 

Sangiovanni, Angelo 229, 393 

Sangro, Bruno 344 

Sanguankeo, Anawin 964 

Sanpajit, Theeranun 1631, 1662 

Sansom, Owen J. 678 

Santana, Rubia A. 1855 

Santangello, Diana 536 

Santantonio, Teresa A. 1802, 1844, 

2003 

Santolaria-Fernandez, Francisco 1342 

Santopaolo, Francesco 203 

Santoro, Michael 1467, 1478 

Santos, Jorge M. 1921 

Santos, Jose Luis 2210 

Santos, Miriam 473 

Santrampurwala, Nishreen 818, 975


Sanyal, Arun J. 52, 105, 107, 157, 

160, 162, 175, 239, 278, 737, 739, 

807, 905, 1309, 1316, 1332, 1428, 

1472, 1521, 2145, 2147, 2153, 

2154, 2163, 2208, 2220, 2235, 

2239, 2249 

Sanz, Ignacio 77 

Sapir, Tamar 1141 

Sapisochin, Gonzalo 53 

Saponaro, Chiara 2209 

Saracino, Giovanna 1210 

Saraf, Neeraj 2245 

Sarangi, Aditya N. 1475 

Sarangi, Vivekananda 1965 

Sarchielli, Erica 882, 902 

Sari, Sinan 1719 

Sarin, Shiv K. 83, 103, 146, 147, 262, 

327, 581, 690, 694, 695, 745, 954, 

1005, 1268, 1328, 1340, 1344, 

1483, 1659, 1663, 1762, 2070, 

2078, 2088, 2092, 2203 

Sarkar, Avik 750 

Sarkar, Monika 4, 1444, 2252 

Sarmati, Loredana 222 

Sarrazin, Christoph 76, 980, 1055, 

1078, 1179, 1559, 1797, 1861 

Sarrecchia, Cesare 222 

Sartor, Ryan B. 1521 

Saruwatari, Junji 1912 

Sasadeusz, Joe 1083, 1161 

Sasaki, Motoko 347, 614, 1901 

Sasaki, Reina 1207, 1628 

Sasaki, Ryo 760, 1464, 1492 

Sasaki, Soichiro 2119 

Sasaki, Yutaka 513, 1912, 1987 

Sasazuki, Takehiko 1632 

Sass, David A. 1276, 1452 

Sastre Turrión, Beatriz 1287 

Sastry, Jagannadha 1030 

Satake, Shintaro 1369 

Satapathy, Sanjaya K. 200, 1212, 

1233 

Satishchandran, Abhishek 496, 512, 

1318, 1330, 1958, 2127 

Sato, Fumiyuki 287 

Sato, Ken 477 

Sato, Masaya 461 

Sato, Mitsuaki 1565 

Sato, Shuichi 2199 

Sato, Shunsuke 1851 

Sato, Takahiro 1116 

Sato, Toshifumi 1771 

Sato, Tsutomu 1984 

Sato, Yasunori 347, 614, 1901 

Sato, Yasushi 1984 

Sato, Matsubara, Misako 517 

Satoh, Hiroaki 551 

Satoh, Takeaki 1159 

Satoskar, Rohit 1101 

Sauer, Peter 618 

Sauer, Vanessa 684, 2103 

Sauerbruch, Tilman 80 

Saunders, Maria 523 

Savard, Christopher 923 

Saviano, Antonio 432 

Savvidou, Savvoula 2012 

Sawada, Koji 927 

Sawadpanich, Kookwan 416 

Sawara, Kei 1528 

Sawhney, Rohit 1482 

Sawinski, Deirdre 871 

Sawle, Ashley 225 

Sax, Paul 1034 

Saxena, Akriti P. 1592 

Saxena, Neeraj K. 1911 

Saxena, Priyanka 1483, 2078 

Saxena, Romil 436 

Saxena, Utsav H. 510, 2116 

Saxena, Varun 471, 1825 

Saxinger, Lynora 641 

Sayiner, Mehmet 153, 892, 1186, 

2173 

Scaglione, Steven J. 532, 559 

Scalone, Luciana 571 

Scanlon, Paul D. 1470 

Scantlebury, Petra 1455 

Scarpi, Emanuela 435 

Scartozzi, Mario 435 

Scatton, Olivier 417 

Scetbun, Jérémy 887 

Schaaf, Sebastian 43 

Schaap, Frank G. 604 

Schaapman, Jelte 2069 

Schaefer, Esperance A. 991, 1015, 

1630, 1644 

Schaeffer, David 1413 

Schaffalitzky de Muckadell, Ove B. 780 

Schaffer, Randolph L. 861, 872 

Schafmayer, Clemens 617 

Scharnagl, Hubert 1726 

Scharpf, Danielle T. 1260 

Scharschmidt, Bruce F. 1467, 1478, 

1485 

Schattenberg, Jörn M. 1949, 2195 

Schaub, Johanna R. 680 

Scheenstra, Rene 1728 

Scheimann, Ann O. 280, 294 

Scheiner, Bernhard 270, 561, 1204 

Schemmer, Peter 1227 

Schewe, Knud 1060, 1081, 1156 

Schiano, Thomas D. 200, 308, 704, 

870, 988, 1095, 1185, 1228, 1250, 

1289, 1337, 1742, 2170 

Schiavon, Leonardo L. 2075 

Schiefke, Ingolf 739 

Schierwagen, Robert 80, 961 

Schiff, Eugene R. 706, 785, 1017 

Schild, Hans-Joerg 1353 

Schillie, Sarah F. 1546 

Schilsky, Michael L. 211 

Schinazi, Raymond F. 1544 

Schinkel, Janke 208 

Schinoni, M I. 595 

Schiöth, Helgi B. 519 

Schippers, Angela 963 

Schlaak, Joerg F. 672 

Schlag, Michael 1179 

Schlansky, Barry 10, 69, 172 

Schlattjan, Martin 1779 

Schlegel, Andrea 499 

Schleicher, Michael 1886 

Schleser, Franziska 808 

Schlicht, Erik M. 607, 610 

Schluckebier, Dominique 1735 

Schmeltzer, Paul A. 17, 1023 

Schmelzle, Moritz 276, 1222, 1394 

Schmerboeck, Bianca 2068 

Schmid, Patrick 1818 

Schmid, Peter 31 

Schmidt, Hartmut H. 684, 2103 

Schmidt, Mark A. 15, 525, 1578, 

1789, 1800 

Schmidt, Monica 14, 533, 2093 

Schmidt, Robin H. 2142 

Schmidt, Warren N. 1100 

Schmidt, Wolfgang E. 2206 

Schmitt, Carolyn 1184 

Schmitt, Timothy 352, 565, 1258, 

1814, 1815 

Schmotzer, Amy R. 1244 

Schmutz, Guenther 1060, 1081, 1156 

Schnabl, Bernd 59, 111, 1319 

Schnee, Matthieu 264, 1072 

Schneider, Kai M. 100 

Schneier, Amanda 1783, 2170 

Schnell, Gretja 705 

Schnickel, Gabriel 1274 

Schnitzler, Paul 1803 

Schober, Andreas 1861 

Schoenbachler, Ben 1574, 1787 

Schölzel, Katrin 1373 

Schork, Nicholas 1431 

Schott, Eckart 37, 252 

Schott, Micah B. 1310 

Schramm, Christoph 76 

Schreder, Alina 2128 

Schreiber, Jonas 2079 

Schreiter, Thomas 1779 

Schroeder, Barbara 1310 

Schrum, Laura W. 1389 

Schrumpf, Elisabeth 258, 819, 833 

Schuch, Anita 1666 

Schuck, Peter 1646 

Schuetze, Marcel 1060, 1081 

Schulze, Ryan J. 1310 

Schumacher, Justin D. 919, 934 

Schuppan, Detlef 43, 119, 183, 950, 

955, 1353, 1368, 1376, 1379, 1385, 

1432, 1434, 1437 

Schuschke, Dale 926 

Schuster, Catherine 1019 

Schütt, jutta 2128 

Schwab, Robert 43 

Schwabe, Christian 713 

Schwabe, Robert F. 98, 1957 

Schwabl, Philipp 149, 270, 475, 561, 

732, 755, 771, 1204, 1525 

Schwartz, Myron E. 253, 254, 380, 

457, 870, 1397, 1742 

Schwartz, Robert E. 229, 679, 1680 

Schwartz Sagi, Liat 1920, 1929 

Schwarz, Kathleen B. 133, 1555, 1694, 

1727, 1729 

Schwarzer, Remy 149, 475, 732, 755, 

771, 1466 

Schwasinger, Emma 1718 

Schweitzer, Nora 444 

Schwimmer, Jeffrey B. 159 

Scian, Romina 2217 

Sciarrone, Salvatore Stefano 1889 

Scioscia, Rosetta 309, 1153 

Sclair, Seth N. 785 

Scoazec, Giovanna 1123 

Scopigno, Tullio 958 

Scott, Gregory 1839 

Scott, Melanie 656 

Scuteri, Alessandra 1888 

Seaberg, Eric C. 1444 

Seal, John 1242 

Seay, Toni 1220


Sebagh, Mylene 1782 

Secchi, Maria Francesca 299 

Secci, Luca 309 

Seebauer, Caroline T. 59 

Seegers, Barbara 1078 

Seeley, Randy J. 916 

Seetharam, Anil B. 1043, 1109, 1145, 

1224, 1241, 1262 

Segato, E. 268 

Segev, Dorry L. 852, 1727, 1729, 

1749 

Segovia-Miranda, Fabian 617 

Seidel, Raphael A. 808 

Seike, Masataka 2031, 2267 

Seipt, Clara C. 1571 

Seki, Akihiro 325, 947 

Seki, Ekihiro 1327, 1345 

Seki, Michiharu 382 

Seki, Shuhji 324 

Sekimoto, Tadashi 1490, 1516 

Seko, Yuya 886, 888, 2202, 2230 

Selby, Warwick 1243 

Sellge, Gernot 100 

Selvapatt, Nowlan 2047 

Selwyn Samraj, Felcy Pavithra 655 

Selzner, Markus 11 

Selzner, Nazia 11, 204, 1012, 1297 

Semela, David 221, 1818, 1884 

Semple, Scott I. 260 

Sen, Bijoya 327 

Sendi, Hossein 1023 

Sendino, Oriol 2084 

Sengupta, Tanya P. 36 

Senju, Takeshi 1157 

Senkerikova, Renata 787 

Sennett, Karen 1455 

Senzolo, Marco 268, 351, 1270, 1889 

Seo, Hyung-Il 1025 

Seo, Jin Seog 424 

Seo, Satoru 402, 1213 

Seo, Wonhyo 1358, 1414 

Seo, Yeon Seok 391, 405, 433, 736, 

1438, 1504, 1605 

Sepp-Lorenzino, Laura 36 

Serejo, Fatima 150, 1876 

Serfaty, Lawrence 105, 162, 251, 701, 

777, 1676, 2145 

Serilmez, Murat 478 

Serji, Badr 1782 

Seror, Olivier 421, 449 

Serper, Marina 122, 1608 

Serra, Giancarlo 309, 1153 

Serrano, Jose 225, 1930 

Sersté, Thomas 1842 

Sese, Pilar 603, 620 

Setchell, Kenneth D. 916, 1695 

Seth, Punit 507 

Seth, Ratanesh K. 962, 2133, 2137 

Seto, Wai-Kay 126, 1545, 1550 

Setoyama, Hiroko 513, 1912, 1987 

Setshedi, Mashiko 1667 

Setsu, Toru 310 

Setze, Carolyn 1084 

Severin, Thomas 260, 753 

Sewell, Justin L. 306 

Seyedkazemi, Star 1041 

Sezaki, Hitomi 1062, 1191, 2020, 

2241 

Sha, Xiaoying 1541 

Shaaban, Mohamed 1271 

Shackel, Nicholas A. 1243, 1264 

Shafi, Muhammad I. 1625 

Shafizadeh, Tracy 1359 

Shah, Anee 2116 

Shah, Apurva 752 

Shah, Hemant 348, 542, 628, 1556 

Shah, Malay B. 854 

Shah, Naina 1482 

Shah, Neeral L. 761 

Shah, Niraj J. 1037, 1074, 1075 

Shah, Omer 1512 

Shah, Puja M. 1501 

Shah, Rohan R. 330 

Shah, Samir R. 578 

Shah, Shawn 580, 1329, 2073 

Shah, Shimul 873 

Shah, Sohela 78 

Shah, Vijay 1315, 1316, 1332, 1355, 

1406, 1947 

Shahinian, Vahakn 395 

Shahoumian, Troy 93 

Shaikh, Haroon A. 521 

Shaikh, Obaid S. 1041, 1753 

Shajari, Shiva 1398, 1416 

Shakado, Satoshi 385, 410 

Shaked, Abraham 5, 853, 868 

Shaker, Mohamed E. 1209 

Shaleh, Hassan M. 1966 

Shalev, Aryeh 2224 

Shan, Hong 1755 

Shang, Jia 1465, 2023 

Shanker, Mihir 784 

Shanmugam, Naresh P. 867 

Shanmukhappa, Shiva K. 823, 1695 

Shannon, Adam 842 

Shao, Benjamin 1469 

Shao, Jian-ying 1647 

Shao, Qing 1536, 1537, 2008 

Shao, Tuo 1305 

Shapiro, David 75, 155, 317, 546, 

562, 609, 625, 628, 644, 645, 1457, 

1458, 1915, 2153, 2154 

Sharif, Khalid 1234 

Sharma, Anuj 1212 

Sharma, Barjesh C. 1473 

Sharma, Dinesh 489, 490 

Sharma, Navneet 259 

Sharma, Shvetank 327, 581, 1344 

Sharma, Suraj 174, 348 

Sharma, Vivek 1753 

Sharma, Yogeshwar 515, 1764, 1917 

Sharpe, Matthew R. 585 

Sharpton, Suzanne R. 852 

Sharr, William 70, 1746 

Shasthry, Saggere M. 581 

Shasthry, Varsha 1328, 1340, 2203 

Shatzel, Joseph 580, 1329, 2073 

Shaughnessy, Melissa 210, 729 

Shaw, David 1083 

Shaw, Kenneth 993 

Shaw, Richard E. 2104 

Shaw, Stanley 2194 

Shawler, Todd 2123 

Shay, Jerry W. 682 

Shayakhmetov, Dmitry 12 

Shearn, Colin T. 921 

Sheen, I-Shyan 1828 

Sheeron, Shawn 609 

Sheiko, Melissa A. 1706, 1708, 1709 

Sheinbaum, Alan J. 488 

Shen, Gong 2265 

Shen, Jianliang 919 

Shen, Ming 12 

Shen, Xi-Zhong 1964 

Sheng, Jifang 2048 

Sheng, Lili 508 

Shepherd, Thomas C. 1445 

Sheppard, Dean 1368 

Sheps, Jonathan 824 

Sherker, Averell H. 133, 213, 1694, 

1922 

Sherman, David I. 791, 800 

Sherman, Morris 174, 348, 542, 851 

Shetty, Kirti 2118 

Shetty, Shishir 1296 

Shi, Aichao 1660 

Shi, Hang 933 

Shi, Hong 131 

Shi, Jiaxiao 1788 

Shi, Tiffany 823 

Shi, Xiang-de 498, 1951 

Shi, Ying 1562 

Shiba, Shunsuke 181, 569, 2131, 

2140, 2234 

Shibata, Hidetaka 1671 

Shibata, Noreene 2109 

Shibata, Toshiya 402 

Shibatou, Toshihiko 456 

Shibolet, Oren 40, 331 

Shiboski, Stephen 1793 

Shiffman, Mitchell L. 94, 106, 205, 609, 

616, 627, 737, 1428, 1813 

Shigefuku, Ryuta 1446 

Shigekawa, Minoru 26, 231, 453, 

1636, 1809, 1982 

Shigematsu, Hirohisa 2031 

Shih, Chao-Jen 2040 

Shih, Mason 1920, 1929 

Shiha, Gamal E. 708, 1076 

Shiina, Shuichiro 461 

Shim, Ju Hyun 446, 448 

Shima, Toshihide 2199 

Shimada, Mitsuo 1983 

Shimada, Noritomo 1097 

Shimada, Shingo 866 

Shimakami, Tetsuro 495, 990, 994, 

1000, 1010, 1348, 1653 

Shimazaki, Tomoe 1672 

Shimizu, Akira 869 

Shimizu, Masahito 657 

Shimizu, Toshiaki 1696 

Shimoda, Michiko 606 

Shimoda, Shinji 640, 2031 

Shimomura, Mayuka 1642 

Shimomura, Yasuyuki 2236 

Shimono, Yoshihiro 466, 770, 781, 

1533, 1669 

Shimosegawa, Tooru 1026, 1648, 

1943, 2167 

Shin, Eui-Cheol 1025, 1650 

Shin, Ji-Hyun 2118 

Shin, Mi-Kyung 1422 

Shin, Seung Kak 1896 

Shinkai, Kazuma 1584 

Shinkai, Masato 1690 

Shinkai, Noboru 1006, 1573, 1599, 

1954 

Shinoda, Masahiro 336 

Shinohara, Fumi 472, 1893 

Shiode, Yuto 669, 1082


Shiota, Goshi 657 

Shiozawa, Hirokazu 2177 

Shirabe, Ken 1983 

Shirafkan, Ali 1991 

Shiraishi, Koichi 314, 383 

Shirasaki, Takayoshi 495, 990, 994, 

1000, 1010, 1014, 1377, 1395, 1653 

Shirasawa, Hiroshi 1628 

Shiratsuki, Shogo 760, 1464, 1492 

Shiri-Sverdlov, Ronit 922 

Shirota, Tomoki 869 

Shivakumar, Pranavkumar 676, 1698 

Shlomai, Amir 875 

Shneider, Benjamin 133, 1694 

Sho, Takuya 287 

Shoji, Hirotaka 476, 1643 

Shomura, Masako 314, 383 

Shoreibah, Mohamed G. 735, 1174 

Shorvon, Philip J. 791, 800 

Shoukry, Naglaa H. 1380 

Showalter, Victor C. 653 

Shrestha, Roshan 200 

Shringarpure, Reshma 239, 2153, 2154 

Shrivastav, Manoj V. 859 

Shrivastava, Shikha 1013, 1029 

Shu, Sally 238 

Shubham, Smriti 690, 694, 1483 

Shui, Xue 510, 2116 

Shukla, Vivek 2118 

Shulga Morskaya, Svetlana 36 

Shulman, Nancy 722, 1039, 1051, 

1065, 1084, 1107 

Shuster, Diana L. 1088 

Si, Won Keun 389 

Si Ahmed, Si Nafa 264 

Sia, Daniela 253, 254 

Siakavellas, Spyros I. 2012 

Sibulesky, Lena 1221 

Siciliano, Massimo 432 

Siddiq, Masood 1872 

Siddiqi, Arif 1872 

Siddique, Asma 248, 250, 1065 

Siddiquee, Zakir 2116 

Siddiqui, Aleem 968 

Siddiqui, Hamda 954 

Siddiqui, Mohammad S. 52, 905, 

1309, 1472, 2163, 2220, 2235, 2249 

Siddiqui, Shaheryar 1513 

Sidharthan, Sreetha 220 

Siebert, Uwe 1089 

Sieghart, Wolfgang 270, 475, 732 

Sievers, Tyson 2006 

Sievert, William 945, 996 

Sigel, Keith M. 1870 

Signoriello, Giuseppe 1543 

Signorovitch, James 711 

Sikaroodi, Masoumeh 905, 1463, 1527 

Sileanu, Florentina E. 275 

Siler, Scott Q. 224, 228 

Silk, Rachel 220 

Sill, Bruce E. 1456, 1461 

Silva, Ismael D. 240 

Silva, Karla 78 

Silva, Lilian M. 84, 2189 

Silva, Luciana C. 2200 

Silva, Marco 1540 

Silva, Raquel S. 473 

Silva, Renato F. 469 

Silva, Telma E. 2075 

Silvain, Christine 3, 206, 1072, 1187, 

2082 

Silveira, Marina G. 1154 

Silvera, Ana 1281 

Silverman, Earl 1713 

Silverman, Gary A. 193, 197 

Simão, Adélia 473 

Simão, André L. 176, 832, 897, 960 

Simerabet, Hayat 816 

Simioni, Paolo 351, 767 

Simmons, Julia 823, 1695 

Simon, Christian O. 1597 

Simon, Karl-Georg 1060, 1078, 1081, 

1156 

Simon, Krzysztof 2001 

Simon, Miguel A. 2032 

Simon-Talero, Macarena 1509 

Simons-Petrusa, Brenna 1790, 1807 

Simpson, Mary Ann 71, 412, 843, 847 

Simpson, Pippa 1914 

Sims, Zayani 92 

Sin, Sui-Ling 1746 

Sinakos, Emmanouil 1126 

Sinclair, Marie 842, 1264 

Sindhi, Rakesh 763 

Sinegre, Thomas 1505 

Singal, Amit G. 371, 418, 548 

Singal, Ashwani 1220, 1249, 1250, 

1253, 1257, 2111 

Singal, Ashwani K. 281, 735, 1316, 

2100, 2114 

Singanayagam, Arjuna 740 

Singh, Ankur 1475 

Singh, Avishek K. 1659 

Singh, Baljinder 259 

Singh, Dharmvir 330, 518, 651, 667, 

668, 671 

Singh, Gurdeep 1817 

Singh, Harsimran D. 1626 

Singh, Shyam 695 

Singh, Tavankit 2182 

Singh, Virendra 259 

Singhal, Pooja 286 

Singhvi, Ajay 1235 

Singla, Manish B. 285, 1866 

Sinha, Rekha 1040 

Sinn, Dong Hyun 422, 423, 485, 1591, 

1960, 2049 

Sinnige, Marjan J. 1633, 2046, 2263 

Siow, Deanna L. 587, 2142 

Sirica, Alphonse E. 1993 

Sirlin, Claude B. 45, 913, 1431, 2150, 

2201 

Sitnik, Roberta 1855 

Sjogren, Maria 285, 1866 

Skaro, Anton I. 71, 563, 843, 847, 849 

Skibba, Kathryn 563, 849 

Skinner, Brianna 2126 

Skinner, Narelle A. 996 

Sklan, Ella H. 331, 672 

Skoien, Richard 1161 

Skorupski, Sharon 1254 

Slagle, Jason M. 529 

Smaïl, Allaoua 554 

Smart, Laura 1260 

Smedile, Antonina 1740, 2209 

Smets, Françoise 1734 

Smith, Abigail 838 

Smith, Alexander 557 

Smith, Bryce 1792 

Smith, Coleman I. 1101 

Smith, Davey M. 1852 

Smith, David B. 993 

Smith, Donna 1048 

Smith, Graham R. 1381 

Smith, Heidi L. 704 

Smith, June 530 

Smith, Mary A. 60 

Smith, Nathaniel 877, 1534 

Smith, Robert 11 

Smith, Robert E. 552 

Smith, Victoria 632, 1367, 1379 

Smits, Loek 2152 

Smuts, Heidi 1667 

Smyrk, Thomas C. 169 

Snead, Nicholas M. 2062 

Sninsky, John J. 131 

Snowball, Mary 125, 1798, 1807 

So, Samuel K. 497, 500 

So-Armah, Kaku 1488 

Soares, Filipa 258 

Soares e Silva, Pedro E. 2075 

Sobajima, Tomoaki 906 

Sobesky, Rodolphe 1237 

Sobhonslidsuk, Abhasnee 1999 

Sobral Dias, Margarida 150 

Söderberg Löfdal, Karin 116 

Soffredini, Roberta 2011 

Sogni, Philippe 373, 1194 

Sohail, Hifsa 1234 

Sohail, Muhammad 968 

Sohda, Tetsuro 385, 410 

Sohn, Joo Hyun 736 

Sohn, Won 485, 1415 

Soin, Arvinder 2245 

Sokal, Etienne M. 1728, 1734 

Sokol, Ronald J. 133, 1687, 1688, 

1694, 1714, 1715, 2134 

Sokolov, Eugene 653 

Solà, Elsa 266 

Solari, Sandra 1217 

Soldevila-Pico, Consuelo 217 

Solé, Cristina C. 266 

Solid, Craig 1867, 1883 

Soliman, Reham 1076 

Solis, Nancy 969 

Solís-Herruzo, José A. 896, 939, 1758 

Solis-Munoz, Pablo 896, 939, 1758 

Sollik, Lisa 1002 

Soltan, Mervat M. 1890 

Soltys, Kyle A. 763 

Somani, Vaibhav S. 752 

Somasundar, Ponnandai 372 

Sommadossi, Jean-Pierre 2266 

Sommer, Lisa 980, 1559 

Son, Byoung Kwan 951 

Sonderup, Mark W. 1667 

Song, Do Seon 1615 

Song, Guisheng 102 

Song, Jinlin 711 

Song, Kyoungsub 686, 1096 

Song, Min-Ae 1553 

Song, Ming 926, 1303 

Song, Sharon 579 

Song, Wei 1554 

Song, Won-Min 364, 2174 

Song, Xiaoling 2151 

Song, Yong 47 

Sonmez, Seval 2057 

Sonnenberg, Amnon 1791


Sonneveld, Milan J. 2001, 2013 

Sonoda, Yuuki 408, 783 

Sood, Siddharth 311, 483, 1225 

Sood, Vikrant 1697, 1725 

Soofi, Madiha E. 1843 

Sookoian, Silvia 2216, 2217 

Soonthornworasiri, Ngamphol 416, 555 

Sophonsritsuk, Areepan 1999 

Sorbo, Maria Chiara 222 

Sorda, Juan A. 2106 

Sorensen, Lisa G. 133 

Soriano, German 583, 596, 1509 

Soriano, Vincent 1180, 1881, 1908 

Sorin, Yushi 1191, 2020, 2241 

Soro-Arnaiz, Ines 930 

Soroka, Carol J. 23, 805, 829 

Soubrane, Olivier 417, 480 

Soulier, Alexandre 999, 1123, 1833 

Sousa, Pedro 661 

Souza, Caroline B. 2072, 2076 

Soza, Alejandro 1161, 1217, 2210 

Sozbilen, Murat 1247 

Sørensen, Michael 768 

Spaan, Michelle 1007 

Spadidea, Panagiota 2094, 2095 

Spadoni, Ilaria 893 

Spaggiari, Mario 850, 1271 

Spahr, Laurent 2082, 2174 

Sparkenbaugh, Erica 81 

Spaulding, Anne C. 1835 

Spearman, C. W. 1667 

Speiser, Jaime L. 1761 

Spengler, Ulrich 37, 252, 1008, 1058, 

1287, 2077 

Sperl, Jan 787 

Spicak, Julius 787 

Spiezia, Luca 351, 767 

Spikes, Leslie 290 

Spindelboeck, Walter 2068 

Spinella, Rosaria 1482 

Spinner, Nancy B. 1693 

Spino, Cathie 1694 

Spirli, Carlo 811 

Spivey, James 200 

Splith, Katrin 276 

Spoletini, Gabriele 1388 

Sponholz, Christoph 808 

Sporea, Ioan 802, 1443, 2004 

Spradling, Philip R. 15, 125, 525, 

1789, 1798, 1800 

Sprague, Andrew G. 36 

Sprinzl, Martin F. 376, 1559 

Squadrito, Giovanni 288, 1476 

Squires, Robert H. 1714 

Sripariwuth, Ekawee 1631, 1662 

Srishord, Alexandrea 534 

Srishord, Indie 153 

Srishord, Manirath 575 

Srivastava, Ankur 793, 1455, 1530 

Srivastava, Siddharth 1473 

Srivastava, Sudhir 371 

Sroczynski, Gaby 1089 

St Clair, Kristina 1175 

St-Laurent Thibault, Catherine 1456, 

1461 

St. Pierre, Timothy G. 798 

Stadlbauer, Vanessa 2068 

Staels, Bart 105, 162, 887, 928, 974, 

2145 

Stahlschmidt, Fabio L. 2248 

Stål, Per 116, 158 

Stambul, Beatrice 1796 

Stamm, Luisa M. 38, 96, 713, 718, 

1034, 1128, 1133, 2265 

Stanco, Marialuisa 148, 266, 292 

Standeford, Holly A. 190, 817, 834, 

835 

Standish, Richard A. 798 

Stanzione, Maria 1837 

Starace, Mario 431, 1543 

Stark, George R. 1025 

Starkel, Peter 1091 

Stättermayer, Albert 297, 732, 1070, 

1137 

Stauber, Rudolf E. 1092, 1885, 2068, 

2069 

Staufer, Katharina 1092 

Staugaard, Benjamin 792 

Stawicka, Agnieszka 1502 

Stedman, Catherine A. 38, 219, 249 

Stefanescu, Horia 741 

Stefano, José Tadeu 240, 2221 

Steffens, Hermann 1170, 1200 

Steinacher, Daniel 977 

Steinberg, Joel L. 1491 

Steindl-Munda, Petra E. 1070, 1137 

Steiner, Sebastian 561, 1204 

Steinmann, Eike 1002 

Stella, Jacquelline 684 

Stelma, Femke 208, 1633, 1995, 2263 

Stepanova, Maria 18, 315, 316, 534, 

1035, 1186, 1460, 2205 

Stephen, Guy R. 202 

Stephenne, Xavier 1734 

Stephens, C. 225, 583, 595, 596 

Stepien, Magdalena 170 

Sterling, Richard K. 52, 1309, 1472, 

1840 

Stern, Nicholas 344 

Stern, Rafael 732, 1070, 1092, 1137, 

1885 

Stern, Sarah 420 

Steuerwald, Nury 1023 

Stevens, James L. 884 

Stewart, Stephen 1103, 1321 

Stewart, Thomas 94 

Sticca, Antonietta 148, 292 

Sticova, Eva 787 

Stieger, Bruno 806 

Stiegler, Philipp 2068 

Stiles, Bangyan L. 1945 

Stine, Jonathan G. 1258, 1501 

Stirling, Kathryn 1298 

Stites, Shana D. 535 

Stivala, Alicia 1095, 1151 

Stockley, Robert 2105 

Stoeckle, Martin 1818 

Stoehr, Albrecht 1176, 1205 

Stoica, Teodora 1114 

Stojakovic, Tatjana 977, 1726 

Stoker, Jaap 2152 

Stokes, Caroline S. 2086, 2253 

Stokes, Scott 1839 

Stokkeland, Knut 116 

Stoll, Barbara 194, 1689 

Stolz, Andrew 225, 1350, 1923, 1930, 

1931, 1932, 1933 

Stolz, Donna B. 197 

Stone, Jack 1794 

Storr, Rachel 2116 

Strachan, Mark W. 2168 

Straley, Stephanie 1825 

Strano, Sabrina 958 

Strassburg, Christian P. 76, 80, 961, 

1008, 1287, 1927, 2077, 2108 

Strasser, Michael P. 1092 

Strasser, Simone I. 609, 1077, 1243 

Strassl, Robert 561 

Straub, Becky 1063 

Strautnieks, S. 1721 

Stravitz, R. Todd 52, 213, 1240, 1309, 

1472, 1761, 1763, 1768, 1914 

Strazzabosco, Mario 571, 743, 811, 

1453, 1950 

Streetz, Konrad L. 963 

Streilein, Robert 2169 

Streinu-cercel, Adrian 1586 

Stremmel, Wolfgang 618, 1411 

Strnad, Pavel 831, 2115 

Strobel, Bastian 1525 

Stroble, Carol 606 

Stroehlein, John R. 130, 682 

Stuart, Alan 2117 

Stuart, Keith E. 412 

Stukenborg, George R. 1501 

Sturdevant, Dan 1029 

Sturm, Ekkehard 1728 

Sturm, Nathalie 1439 

Stypinski, Daria 730, 731 

Su, Chien-Wei 365, 409, 439, 751, 

757, 759, 1639, 2050 

Su, Christopher 1606 

Su, Danmei 904 

Su, Feng 1211 

Su, Grace L. 418 

Su, Jie 1411 

Su, Lishan 1620 

Su, Shicheng 1423 

Su, Tung-Hung 1996 

Su, Wei-Wen 1996 

Su, Wen-Pang 2040 

Su, Xiaoping 130, 682 

Su, Ying-Hsiu 1554 

Su, Zhuang 2035 

Suarez, Emilio 2032 

Suarez, Yajaira 65 

Suazo, Jose 2210 

Subajima, Tomoaki 1642 

Subramaniam, Nathan 975 

Subramanian, Mani 241, 616, 624, 

632, 737, 739, 746, 777, 1428, 

1435, 1442, 1551, 1557, 1563, 

1651, 2004, 2014, 2015, 2025, 

2037, 2043, 2052, 2059, 2149, 2239 

Subramanian, Ram M. 82, 570 

Subramanian, Savitha 923 

Suchy, Frederick J. 809, 1688 

Suda, Goki 287, 1672, 1946 

Suda, Jo 2135 

Suda, Takahiro 26, 1809, 1982 

Suda, Tsuyoshi 699 

Suddle, Abid 646, 1113, 1744 

Sudo, Tomoko M. 1378 

Sue, Paul K. 560 

Suehiro, Mitsuhiko 2030 

Suemizu, Hiroshi 1635 

Suemura, Shigeki 1635, 1809, 1982 

Sueoka, Hideaki 1420 

Sugawara, Kayoko 1059, 1064, 1474, 

1486


Sugihara, Takaaki 1499 

Sugimoto, Katsutoshi 1047 

Sugimoto, Kazushi 1020 

Sugimoto, Rie 1159 

Sugimoto, Ryosuke 584, 1500 

Sugimoto, Satoru 901 

Sugiura, Tokio 135 

Sugiyama, Kazuo 569 

Sugiyama, Masaya 476, 1138, 1614, 

1632, 1643, 1658, 1673, 1795, 

2158, 2193 

Sugiyama, Nao 1003 

Suguness, Arvind 1831 

Suh, Dong Jin 446 

Suh, Jeong Ill 2038 

Suh, Kyung-Suk 400, 474, 853, 1277 

Suh, Sang Jun 391, 405, 1897 

Suh, Suk-Won 853, 1278 

Suhy, David 2258 

Suk, Fat-Moon 1996 

Suk, Ki Tae 736, 1515, 1957 

Sukeepaisarnjaroen, Wattana 1631, 

1662 

Sukriti, Sukriti 327, 1344, 2078 

Suksawatamnuay, Sirinporn 1631, 1662 

Sulfab, Maisoun 1359 

Sulkowski, Mark S. 91, 94, 205, 210, 

219, 250, 700, 702, 703, 712, 716, 

1035, 1039, 1057, 1749 

Sullivan, Danielle 1161, 1198 

Sultan, Khaleel H. 1147, 1188 

Sultan, Maya 1685 

Sultanik, Philippe 1194 

Sumazaki, Ryo 1690 

Sumida, Yoshio 886, 888, 901, 1980, 

2156, 2158, 2202, 2207, 2230 

Sumiyoshi, Hideaki 1369 

Sumpter, Colin 113 

Sun, Yameng 374 

Sun, Bing 1149 

Sun, Chao 2125 

Sun, Haibo 1149 

Sun, Jing 1580, 1620 

Sun, Mengxi 186 

Sun, Qian 656, 1335 

Sun, Runbin 919, 934 

Sun, Xiaochen 229, 254, 364, 1392, 

1940, 2174 

Sun, Xiaofeng 1394 

Sun, Ying 77, 639 

Sun, Ying S. 945 

Sunagozaka, Hajime 1348 

Sundaram, Shikha 137, 1709, 1715, 

1737, 1738 

Sundaram, Vinay 566, 1279, 1479, 

1512, 2083 

Sung, Pil Soo 1025 

Superbia, Marcel 1484, 1494, 1495 

Supper, Paul 1525 

Surana, Pallavi 779 

Suriguga, Suriguga 1427, 1434 

Susser, Simone 980, 1055, 1559 

Sussman, Norman L. 9, 1761 

Suto, Hiroyuki 944, 1682 

Sutton, Angela 362 

Sutton, Harry 134 

Suzuki, Ayako 540, 596 

Suzuki, Fumitaka 707, 1062, 1191, 

2020, 2241 

Suzuki, Harukazu 657 

Suzuki, Hitoshi 551 

Suzuki, Kazuyuki 1528 

Suzuki, Kunio 2213 

Suzuki, Michihiro 1446 

Suzuki, Mitsuyoshi 1696 

Suzuki, Shoko 1115, 1192 

Suzuki, Tetsuro 1393 

Suzuki, Yasuaki 2207 

Suzuki, Yoshiyuki 1062, 1191, 2020, 

2241 

Suzuki, Yuhei 477 

Suzuki, Yuichiro 1565 

Suzumura, Kazuhiro 1420 

Svarovskaia, Evguenia S. 91, 219, 249, 

713, 1168 

Svegliati Baroni, Gianluca 21, 355 

Sverdlov, Deanna 1368, 1379 

Svicher, Valentina 203 

Swain, Mark 1290 

Swain, Telisha 1313 

Swallow, Elyse 711 

Swaney, James 141 

Swanson, Herbert 313, 537, 546, 562 

Swanson, William M. 1117, 1132 

Swearingen, Christopher J. 1914 

Swearingen, Dennis 725 

Sweeney, William E. 1718 

Swenson, Eugene S. 702, 706, 716, 

726 

Swet, Jacob H. 653 

Swiderska, Aleksandra 1502 

Swiderska-Syn, Marzena 66, 1386, 

1715, 2143 

Swierczewska, Magdalena 115 

Sydor, Svenja 1779 

Sykes, Megan 2129 

Symonds, Bill 1094 

Symons, Julian A. 993 

Syn, Wing-Kin 66, 163, 1978, 2143 

Sypsa, Vana 2012 

Szabo, Gyongyi 109, 180, 496, 512, 

1016, 1318, 1330, 1958, 2127, 2198 

Szabo, Shelagh M. 1139, 1152 

Szanda, Gergo 1374 

Szenborn, Leszek 2043 

T. Dogan, Omer 2219 

Tabak, Fehmi 241, 1586, 2001, 2013, 

2014 

Taber, David J. 1272 

Tabernero, David 1021, 1674 

Tabibian, James H. 605, 643 

Tabrizian, Parissa 851 

Tachi, Yoshihiko 1834 

Tachlytski, Irina 1685 

Tack, Kenneth 2257, 2259, 2260 

Tacke, Frank 100, 961, 1756 

Tada, Seiya 1159 

Tada, Toshifumi 344 

Taddei, Tamar H. 16, 743, 1899 

Tadrous, Paul 791, 800 

Tafarel, Jean R. 2248 

Tagawa, Kazumi 382 

Taguchi, Y-h 429 

Tahata, Yuki 1195, 1197, 1201, 2213 

Tai, Andrew W. 27 

Tai, Chi-San 1705 

Tai, Ming-Hong 1942 

Taibi, Chiara 1751 

Tajima, Kazuki 336 

Tajiri, Kazuto 1599 

Tak, Won Young 363, 403, 426, 749, 

2025, 2028, 2038 

Takabatake, Hisashi 325 

Takabatake, Riuta 1377, 1395, 1968 

Takada, Hitomi 1115 

Takagi, Hitoshi 477 

Takaguchi, Koichi 1097 

Takahashi, Atsushi 551 

Takahashi, Kazuaki 1778 

Takahashi, Kazuhiro 1274, 2031 

Takahashi, Kazuto 944 

Takahashi, Masahiko 2177 

Takahashi, Takeshi 1635 

Takahashi, Tetsuyuki 1981 

Takahira, Sachiko 314 

Takai, Satoshi 1648 

Takaki, Akinobu 339, 2236 

Takaki, Yugo 1724 

Takakura, Kazuki 2074 

Takamatsu, Shinji 906, 1642, 2156 

Takamatsu, Yuki 1655 

Takami, Taro 329, 760, 1447, 1464, 

1492 

Takami Lageborn, Christine 116 

Takamura, Masaaki 310, 2165 

Takamura, Toshinari 144, 990, 1010, 

1348, 1377, 1968 

Takase, Masaru 820 

Takashi, Niizeki 455 

Takashima, Tomoyuki 466, 770, 781, 

1533, 1669, 1975 

Takata, Kazuhide 385, 410 

Takata, Ryo 466, 770, 781, 1533, 

1669 

Takatori, Hajime 699 

Takayama, Kazuo 1690 

Takayama, Tadatoshi 407 

Takebe, Takanori 676 

Takeda, Kazuyoshi 2119 

Takegoshi, Kai 144, 1377, 1395, 1968 

Takehara, Tetsuo 26, 97, 227, 231, 

453, 669, 906, 1082, 1195, 1197, 

1201, 1357, 1635, 1636, 1642, 

1661, 1809, 1974, 1982, 1985, 

2130, 2136, 2156, 2213 

Takei, Hajime 804, 1696 

Takei, Yoshiyuki 584, 1020, 1500 

Takeiri, Masatoshi 1918 

Takemoto, Kenji 1918 

Takemura, Shigekazu 429, 1748 

Takeshita, Eiji 2146 

Taketani, Hiroyoshi 886, 888, 901, 

2230 

Taketomi, Akinobu 476, 866, 1632 

Takeuchi, Takahito 1724 

Takeuchi, Yasuto 2236 

Takeyama, Yasuaki 385, 410 

Takigawa, Atsuo 26, 453, 1809, 1982 

Takikawa, Hajime 1757 

Takikawa, Yasuhiro 1528 

Takimoto, Rishu 1984 

Takkenberg, Bart 1633, 1995, 2005, 

2046 

Takyar, Varun K. 779, 1498 

Talal, Andrew H. 332, 1823, 1841 

Talaty, Jennifer E. 725 

Taleb, Eman 1118 

Taliani, Gloria 222, 983, 1802, 1844 

Tallis, Caroline 1077 

Talmat, Nabila 1847


Talwani, Rohit 251, 715 

Tam, Edward 1051 

Tamai, Tsutomu 799, 937, 1421, 1838 

Tamaki, Nobuharu 1115 

Tamberi, Stefano 435 

Tamburrino, Domenico 1388 

Tamè, Maria Rosa 1751 

Tameda, Masahiko 1020 

Tamhane, Ashutosh 1804 

Tamko-Mella, Ghislaine Flore 1833 

Tamori, Akihiro 429, 1125, 1748, 

1824, 1983 

Tamura, Yasuaki 1375, 1378 

Tan, Amy 870 

Tan, Bingyan 454 

Tan, Brandon 916 

Tan, Chee-Kiat 451 

Tan, Corey 819 

Tan, Damien 167 

Tan, Hua 993 

Tan, Poh Seng 364 

Tan, Youwen 2048 

Tana, Michele M. 306 

Tanabe, Kazutaka 1213, 1918 

Tanabe, Kenneth K. 957, 1938 

Tanabe, Minoru 406 

Tanahashi, Toshihito 429 

Tanaka, Eiji 1632, 1673 

Tanaka, Hajime 613 

Tanaka, Hiroki 927 

Tanaka, Hironori 1975 

Tanaka, Kathryn 1333 

Tanaka, Motohiko 513, 1912, 1987 

Tanaka, Sadao 456 

Tanaka, Saiyu 2156, 2202, 2230 

Tanaka, Satoshi 97, 231, 453, 669, 

1082, 1357, 1661, 1985, 2130, 2136 

Tanaka, Shingo 1984 

Tanaka, Shinji 406, 1980 

Tanaka, Shogo 1748 

Tanaka, Takashi 385, 410 

Tanaka, Takuji 144, 1377, 1395, 1968 

Tanaka, Yasuhito 1006, 1018, 1026, 

1097, 1138, 1448, 1573, 1599, 

1618, 1640, 1655, 1983, 2063 

Tanaka, Yasunobu 1433 

Tanaka, Yasuo 461 

Tanaka, Yujiro 1018 

Tanaka, Yuki 1159 

Tandoi, Francesco 1, 1740 

Tandon, Puneeta 82, 570, 742 

Tanfin, Zahra 816 

Tang, Alister L. 1121 

Tang, Hong 1562 

Tang, Weiliang 1611 

Tang, Yilang 183, 1353 

Tange, Kazuhiro 524 

Tangkijvanich, Pisit 2036, 2063 

Taniai, Makiko 1326 

Taniguchi, Eitaro 233 

Tanikawa, Ken 1690 

Tanikawa, Tomohiro 2030 

Tanikella, Rajasekhar 1513 

Taniki, Nobuhito 181, 569, 2131, 

2140, 2234 

Tanimoto, Hironori 2031 

Tanné, Florence 264 

Tanno, Federico C. 595 

Tanoue, Yasushi 505 

Tantipanichtheerakul, Supot 1631, 1662 

Tanwandee, Tawesak 467, 1631, 1662 

Tanwar, Sudeep 793, 1455, 1530 

Tao, Junyan 1948 

Tao, Shiqi 1865 

Tao, Sijia 1544 

Tapper, Elliot B. 566, 1222, 1477, 

2196, 2197, 2227 

Tariciotti, Laura 203 

Tarocchi, Mirko 949, 1963 

Tarr, Phillip 2091 

Tartaglione, Erica 803 

Tartof, Sara Y. 1135 

Tashiro, Shigeki 1159 

Tashiro, Taku 1584 

Tate, Janet 1488 

Tateishi, Ryosuke 461 

Tateno, Chise 1150, 1670, 1679, 2120 

Tateyama, Masakuni 1912, 1987 

Tatosian, Daniel A. 707 

Tatsumi, Kohei 81 

Tatsumi, Tomohide 26, 97, 231, 453, 

669, 1082, 1195, 1197, 1201, 1357, 

1635, 1636, 1661, 1809, 1982, 

1985, 2130, 2136, 2213 

Taura, Kojiro 402, 1213, 1918 

Taura, Naota 952, 1671 

Tawara, Seiichi 26, 453, 1635, 1809, 

1982 

Tawdrous, Monika 2068 

Taylor, Amy E. 1695 

Taylor, Avril 1794 

Taylor, James 718 

Taylor, Ryan 1291, 1761, 1814, 1815 

Taylor, Sandra L. 606 

Taylor, Simeon 141 

Taylor, Stephanie 1309 

Taylor, Thom 1048 

Taylor, William R. 169 

Tayyab, Ghias Un Nabi 1872 

Tchaikovskaya, Tatyana 515 

Te, Helen S. 271, 539, 544, 553, 579, 

1101, 1229 

Teachenor, Olga 1146, 1199 

Teckman, Jeffrey 121, 1694 

Tedgui, Alain 887 

Telese, Andrea 337, 1388 

Tellatin, S 268 

Teltsch, Dana Y. 1175 

Tempesti, Sara 949, 1963 

Temple, Luisa 78 

Temple, Sarah 357 

Ten Have, Gabriella A. 671, 1523 

ten Kate, Fiebo J. 1590 

Teng, Kun-Yu 493 

Teng, Ruei-Dun 1639 

Tenti, Elena 435 

Teo, Chong-Gee 2126 

Teoh, Narci C. 55, 230 

Teperino, Raffaele 1386 

Teperman, Lewis W. 200 

Terai, Shuji 310, 1447, 2165 

Teranishi, Yuga 592, 1125 

Terao, Naoko 906 

Terashima, Takeshi 370, 392, 1653 

Terra, Carlos 84, 2189 

Terrault, Norah 94, 200, 236, 239, 

471, 1034, 1084, 1106, 1259, 1560, 

1598, 1825, 2153, 2154, 2161, 

2220, 2252 

Terrin, Norma 1592 

Teruki, Miyake 524 

Teshale, Eyasu H. 15, 525, 1789, 1800 

Testro, Adam 1225 

Teta-Bissett, Monica 494, 511 

Teterina, Anastasia 2238, 2254 

Teti, Elisabetta 222 

Teuber, Gerlinde 1170, 1200, 1861 

Teuber, Peter 278 

Tevar, Amit D. 216 

Thabut, Dominique 263, 744, 1510 

Thacker, Leroy 82, 570, 1463 

Than, Tin A. 590 

Thanapirom, Kessarin 360, 1631, 1662 

Thandassery, Ragesh B. 1843 

Thapaliya, Samjhana 651, 667, 668, 

671, 1308 

Tharinger, Hugo 2009 

Tharwa, El-Sayed S. 1862, 1879 

Thasler, Wolfgang E. 520, 1684 

Theise, Neil D. 681, 685, 2214 

Therapondos, George 200, 1242 

Therneau, Terry M. 6, 397 

Thevenon, Sylvie 1162 

Thevenot, Thierry 145, 264, 554, 1036, 

1187, 2082 

Thi, Emily P. 2007, 2034, 2062 

Thibault, Vincent 786, 1808 

Thiele, Maja 773, 780, 789, 792, 

1440 

Thimme, Robert 1666, 1803 

Thirumurthy, Harsha 1063 

Thogaripally, Suneetha 1804 

Thoma, Christian 2183 

Thomas, Emmanuel 785, 1017 

Thomas, Howard 1551 

Thomas, Peter 1969 

Thomas, Richard C. 2226 

Thomes, Paul G. 1323, 1347, 1389 

Thomopoulos, Konstatinos 2094, 2095 

Thompson, Alex J. 106, 249, 442, 627, 

703, 996, 1077, 1551, 1557, 1563, 

1651, 2037, 2262 

Thompson, Richard J. 1721, 2105 

Thomsen, Karen Louise 908, 1300 

Thomson, Angus W. 338 

Thomson, Mary 576 

Thongsawat, Satawat 1631, 1662 

Thonig, Antje 1519 

Thorburn, Douglas 73, 76, 413, 634, 

1216 

Thorgeirsson, Snorri S. 677 

Thuluvath, Paul J. 82, 570, 739 

Thung, Swan N. 254, 506, 870, 1337 

Thurner, Lorenz 463 

Thursfield, Vicky 442 

Thuy, Tuong Thi Van 191 

Tian, Dean 328 

Tian, Hui 198, 1986 

Tian, Jijing 1285 

Tian, Lei 1396 

Tice, Ashley B. 166 

Tieleman, Madelon 68 

Tien, Phyllis 2252 

Tierney, Amber 1101, 1184 

Tiirikainen, Maarit 1553 

Tijeras-Raballand, Annemilaï 417 

Tikhanovich, Irina 1291 

Tikhonova, Natalia 721 

Tillman, Bryan 216 

Tillman, Holly 213, 1761, 1768


Tillmann, Hans L. 540, 1922, 1931 

Timm, Joerg 672 

Ting Zhang, Ting 1939 

Tiniakos, Dina 652 

Tinti, Francesca 983 

Tiribelli, Claudio 1339, 1346, 2098 

Tiro, Jasmin A. 548 

Tisdale, John F. 1498 

Tisone, Giuseppe 203, 1751 

Titos, Esther 2087 

Tizzard, Sarah 1704 

Tobita, Hiroshi 2199 

Toda, Nobuo 382, 2240 

Todus, Robin 1172 

Toffanin, Sara 253 

Togawa, Takao 135 

Togayachi, Akira 1448 

Tokoro, Masanori 2267 

Tokumoto, Yoshio 788, 1293, 2146 

Tokunaga, Katsushi 1138, 1632 

Tokushige, Katsutoshi 1326, 2207 

Toledano, Ronen 2224 

Tolenaars, Dagmar 604, 827 

Toli, Barbara 1126 

Tomasi, Maria Lauda 594 

Tomasiewicz, Krzysztof 1582 

Tominaga, Kentaro 310 

Tomita, Kyoko 895 

Tomiyama, Takashi 613, 639 

Tomkoetter, Lena 1282 

Tona, Francesco 268 

Tonascia, James 160, 2150, 2220 

Tonello, Silvia 775 

Tong, Myron 1165 

Tong, William 1571 

Tonnu-Mihara, Ivy 1110 

Tonon, Marta 148, 292, 767, 775 

Tontini, Gian Eugenio 1506 

Topal, Baki 171 

Topic, Nina 538 

Torbeyns, Anne 702 

Tordjmann, Thierry 816 

Toribio, Wilma 1870 

Toriguchi, Kan 1918 

Torii, Nobuyuki 1326 

Torimura, Takuji 233, 304, 399, 455, 

681, 688, 918, 1819, 1990, 2158 

Tornai, David 2090 

Tornai, Istvan 2090 

Tornai, Tamas 2090 

Torok, Natalie J. 1285, 2179 

Torre, Aldo 1471, 1508, 2223 

Torrens, Maria 1509 

Torres, Dawn M. 285, 1866, 2239 

Torres, Harrys A. 1030, 1160 

Torres, Javiera 969 

Torres Hernandez, Alejandro 1282 

Torres-Capelli, Mar 930 

Torres-Gonzalez, Edilson 2142 

Torzilli, Guido 602 

Toshimori, Akiko 524 

Toshiro, Takezaki 1421 

Tosti, Maria Elena 1802, 1844 

Tosti Guerra, Cristina 1410 

Tosun, Selma 1622, 1623 

Toutouza, Marina G. 1489 

Towner, William J. 205 

Townsend, Mary L. 1142 

Townshend-Bulson, Lisa J. 1790, 1798, 

1807 

Toyoda, Hidenori 344 

Toyota, Joji 1116, 2065 

Tozun, Nurdan 558, 1822 

Trabucchi, Michele 987 

Trabut, Jean-Baptiste 1102 

Tracy, Russel 2228 

Traetow, Daniel 1260 

Trakroo, Sushrut 1122 

Tran, Albert 3 

Tran, Chris 36 

Tran, Lien 269 

Tran, Patricia 316 

Tran, Tram T. 249, 1479 

Tran, Viet Chi 1845 

Tran Minh, Margherita 1239 

Trapero, Maria 1511 

Trauner, Michael 76, 149, 270, 297, 

475, 561, 609, 662, 732, 755, 771, 

977, 1070, 1092, 1137, 1204, 1466, 

1525, 2164 

Trautwein, Christian 100, 828, 831, 

961, 963, 1045, 1756, 1924, 2115 

Trawick, Bobby N. 1209 

Traxinger, Brianna 1692, 1699 

Trebicka, Jonel 80, 773, 961 

Treeprasertsuk, Sombat 360, 416, 555 

Trehanpati, Nirupma 327, 695, 1344, 

1659, 1663, 1762, 2088 

Tremblay, Mélanie 51, 1523 

Trembling, Paul M. 793, 1530 

Trenell, Michael I. 2183 

Trepo, Christian 1162 

Trépo, Eric 178, 2079 

Treviño, Ana 1180 

Trevisani, Franco 355, 356 

Triantos, Christos K. 2094, 2095 

Trinacty, Connie M. 15, 525, 1578, 

1789, 1799, 1800 

Trindade, Alexandre 897 

Trindade, Miguel 1876 

Trinh, Huy N. 123, 207, 368, 1182, 

1549, 1561, 2015, 2025, 2052 

Trinh, My My 2262 

Trinh, Roger 722, 1051 

Trinh, Thu Le 1967 

Tripathi, Dinesh M. 1518 

Tripathi, Rakesh 705 

Tripodi, Gianluca 1654 

Tripon, Simona 744, 1510 

Trivedi, Palak J. 73, 76, 634 

Trooskin, Stacey B. 1864, 1880 

Trujillo, Ruby 1121 

Trunecka, Pavel 787 

Trychta, Kathleen 323 

Tsai, Ming-Chao 1942 

Tsai, Naoky CS C. 1046, 1108, 1121, 

1553 

Tsai, Shannon 195 

Tsai, Yi-Shan 2264 

Tsang, Owen Tak Yin 2025 

Tsang, Rebecca 532, 559 

Tschernig, Thomas 1299, 1385 

Tschudy-Seney, Benjamin 685 

Tschuor, Christoph 665 

Tse, Yee-Kit 349 

Tseng, Kuo-Chih 1996 

Tseng, Po-Lin 1873, 1942 

Tseng, Tai-Chung 1996 

Tsianos, Epameinondas 801 

Tsianou, Zoi 801 

Tsien, Cynthia 667 

Tsipouras, Markos 801 

Tsiriga, Athanasia 1489 

Tsochatzis, Emmanuel 413, 764, 1455 

Tsubouchi, Eiji 2146 

Tsubouchi, Hirohito 937, 1421, 1757 

Tsuchida, Takuma 1392 

Tsuchiura, Takayo 1632 

Tsuchiya, Hiroyuki 1336 

Tsuchiya, Kaoru 1115 

Tsuds, Yasuhiro 966 

Tsuge, Masataka 472, 663, 986, 1056, 

1150, 1627, 1645, 1649, 1670, 

1681, 1895, 1944, 2120 

Tsujii, Masahiko 1197, 2213 

Tsukamoto, Hidekazu 112, 682, 904, 

1936, 2118 

Tsunematsu, Seiji 287, 1946 

Tsuneyama, Koichi 1981 

Tsung, Allan 184 

Tsunoda, Tomoyuki 164 

Tsuruga, Yosuke 866 

Tsuruya, Kota 314, 383, 697, 2177 

Tsutsumi, Susumu 1006, 1599 

Tsutsumi, Takeya 912 

Tsuzaki, Ryuichiro 339 

Tsuzura, Hironori 1851 

Tucker, Edwin 1920, 1929 

Tucker, Joseph D. 577 

Tujios, Shannan R. 290, 1766 

Tuma, Dean J. 589, 1323, 1347, 1969, 

2139 

Tumas, Daniel 1359 

Tummala, Swetha 201, 1745 

Tunnicliffe, Elizabeth M. 2190 

Tunõn, María Jesús 1398 

Tur-Kaspa, Ran 875 

Turan, Ilker 246, 1247, 1739 

Turcios, Lilia 673 

Turmelle, Yumirle P. 133, 1694 

Turner, Samuel 89, 1090, 1852 

Turner, Scott M. 1425 

Turnquist, Heth R. 184 

Turquet, Jeid 2105 

Turturro, Francesco 1160 

Tuzun, Ali 2033 

Tyndall, Mark 86, 543 

Tzallas, Alexandros T. 801 

Tzarnas, Stephanie 1864, 1880 

U, Rebecca Y. 177 

Uchida, Daisuke 2236 

Uchida, Sawako K. 429, 1125, 1824 

Uchida, Shinjiro 783, 1600 

Uchida, Takuro 663, 986, 1150, 1649, 

1670, 1681, 1858, 1895, 1944, 2120 

Uchida, Yoshihito 1059, 1064, 1577, 

1757 

Uchikoshi, Manabu 1302 

Uchino, Koji 461 

Uchiyama, Akira 226, 917, 1771 

Uddin, Muazzam 1872 

Udo de Haes, Joanna 208, 2263 

Udoh, Uduak S. 1313 

Udompap, Prowpanga 88, 1208, 2155 

Ueda, Keiji 1642 

Uehara, Daisuke 477 

Uehara, Hisanori 1981 

Uehara, Takahide 524 

Ueland, Joseph 709 

Uemoto, Shinji 402, 1213, 1918, 1976


Uemura, Hayato 1064 

Ueno, Takato 688, 918 

Uesaka, Katsuhiko 820 

Ueyama, Misuzu 794, 1795, 2158, 

2193 

Ulivi, Paola 435 

Ullah, Naeem 1275 

Ullmer, Christoph 816 

Um, Soon Ho 391, 433, 736, 1438, 

1504, 1605 

Umbro, Ilaria 983 

Umeda, Rumiko 2177 

Umemura, Atsushi 232, 886, 888, 901, 

1980 

Umemura, Machiko 1672 

Umeno, Yumi 1990 

Umetsu, Teruyuki 1026, 1648, 1943, 

2167 

Ungerer, Jacobus 784 

Ungerleider, Nathan 686 

Unler, Gülhan K. 2206 

Unser, Ariel 52, 1463, 1472, 1491, 

1527 

Upala, Sikarin 964 

Uprichard, Susan L. 982 

Urabe, Ayako 1195, 1201, 2213 

Urata, Noriyo 2030 

Urban, Thomas J. 225, 1930, 1933 

Urbanek, Petr 1487, 1678 

Urbani, Luca 337, 1388 

Uribe, Claudia L. 1050 

Uriel, Alison J. 1112 

Urraro, Teresa 1190 

Urrunaga, Nathalie H. 1911 

Urrutia, Raul A. 692 

Ursic-Bedoya, José 816 

Urushihara, Naoko 1378 

Uschner, Frank E. 80, 961 

Uslan, Mustafa I. 846 

Usui, Akihiro 1993 

Usui, Shingo 569 

Uto, Hirofumi 799, 937, 1421, 1838 

Utrankar, Amol 1223 

Utsumi, Masashi 339 

Utsumi, Teruo 1322, 1526 

Utsunomiya, Hiroki 2146 

Utsunomiya, Tohru 1983 

Uyama, Naoki 188, 1420 

Vadalà, Domenica 288 

Vaddady, Pavan 731 

Vadnal, Prashant 118 

Vaillant, Andrew 31 

Vaishnav, Joban 1255 

Valantin, Marc-Antoine 568, 1093 

Valdeolivas, Tatiana 2072, 2076 

Valderama, Adriana 16 

Vale, Ana 1540 

Vale, Luke 636, 1457 

Valenti, Luca 973, 2187 

Valentin, Nelson 282 

Valla, Dominique C. 772, 1288, 2148 

Vallet-Pichard, Anaïs 95, 1158, 1194 

Vallier, Ludovic 258 

Valveri, Vincenza 2011 

Van Allen, Jessica 1770 

van Bömmel, Alena 245 

van Bömmel, Florian 245, 1559, 1594, 

1638, 2003, 2012 

van Buuren, Henk R. 68, 73, 302, 601, 

634 

van Campenhout, Margo J. 1585, 1586 

van de Garde, Martijn D. 2122 

van de Graaf, Stan F. 637 

van de Laar, Thijs J. 1846 

van den Berg, Aad P. 68, 302 

Van Den Berg, Machteld 167 

van den Boom, Rivka 1238 

Van Der Graaff, Denise 976 

van der Helm, Jannie 1846 

van der Mark, Vincent 492 

van der Meer, Adriaan J. 350, 445, 

1166, 1590, 2255 

Van Der Merwe, Schalk 171 

van der Net, Guido 2122 

Van der Ploeg, Lex H. 143 

van der Poorten, David 441 

van der Ree, Meike H. 208, 2005, 

2255, 2263 

van der Reijden, Johan 2069 

van der Valk, Marc 37, 208, 252, 1058 

Van der Ven, Peter F. 80 

van Dijk, Remco 492 

Van Dooren, Gino 1163, 1179 

van Dort, Karel A. 1633, 2005 

van Erpecum, Karel J. 609 

Van Eygen, Veerle 39 

van Gaal, Luc 928 

van Gorp, Patrick 922 

van Gulik, Thomas 637 

van Hoek, Bart 68, 302, 1049, 1238, 

2069 

Van Hul, Noemi 678 

Van Itallie, Christina 20 

van Leeuwen, Ester M. 1633 

van Liempd, Sebastiaan 953 

Van Natta, Mark L. 107, 157, 236, 

2150 

Van Nieuwkerk, Carin M. 302, 1995 

van Nuenen, Adrianus C. 2255 

van Oord, Gertine 1007, 1586 

Van Remoortere, Peter 39 

van Scherpenzeel, Monique 2102 

van Seggelen, Wouter 1112, 1852 

van Silfhout, Joanne J. 302 

Van Steenbergen, Werner 171 

Van Steenkiste, Christophe 1091 

Van Vlierberghe, Hans 1049, 1067 

van Vliet, Andre 208, 2263 

Van Wagoner, David R. 518, 651, 667 

Van Wilgenburg, Bonnie 1298 

Vanatta, Jason 1212, 1233 

Vanderhoff, Aaron M. 1151 

Vandevoorde, Ann 39 

Vanlemmens, Claire 1187 

Vannelli, Gabriella Barbara 882, 902 

Vanslembrouck, Ragna 171 

Vanveuren, Charlotte 2222 

VanWagner, Lisa B. 563, 849, 1235 

Vanwolleghem, Thomas 797, 1637, 

2122 

Vaquero, Javier 1295, 1520 

Varaut, Anne 1123 

Varela, Maria 378 

Varela, Marta 953, 2244 

Vargas, Hugo E. 82, 248, 250, 570, 

1038, 1106, 1124 

Vargas, Juan 78 

Vargas, Victor 609, 2069 

Varghese, Rony 2162, 2186 

Varier, Raghu 1702 

Varma, Sharat 1728, 1734 

Varshney, Aditi 1659 

Varunok, Peter 1065 

Vasconcelos, Ana Clara 2237 

Vasilescu, Alexandra 280 

Vasilieva, Larisa E. 1489, 2096 

Vasudevan, Madavan 695, 1659 

Vatcheva, Kristina 1424 

Vatsalya, Vatsalya 1334 

Vavassori, Sara 393 

Vega, Maricruz 597, 1932 

Veidal, Sanne S. 978 

Veitsman, Ella 1032 

Velame, Daniela 2237 

Veldt, Bart J. 445, 1166 

Vella, Stefano 1153, 1802, 1844 

Vellicce, Alejandra 2106 

Vellozzi, Claudia 1574, 1787, 1792 

Vellucci, Vincent 709 

Velosa, José F. 150, 1876 

Veloso, Julio M. 1256 

Venditti, Mario 2067 

Venkat, Nandini 2116 

Venkatesan, Chapy 575 

Venkatesh, Anu 364, 2174 

Venter, Julie 190, 621, 817, 834, 835, 

929, 1317, 1349 

Ventura-Cots, Meritxell 380, 438, 457, 

1509 

Venugopal, Senthil K. 1664 

Vergani, Diego 300, 608, 1704 

Vergniol, Julien 450, 1439, 2181 

Verhaag, Esther M. 591 

Verheij, Joanne 637, 1995, 2046, 

2152 

Verhey, Elke 2002 

Verheyen, Fons 922 

Verkade, Henkjan J. 1728 

Verlinden, Wim 797 

Verma, Manisha 530, 535, 1932 

Verma, Neha 1451 

Verma, Suman 646, 1098, 1113, 

1117, 1132, 1173, 1216, 1588, 2054 

Verma, Vikas K. 1315, 1406 

Verma, Yogita 1268 

Vermehren, Johannes 1055, 1078, 

1797 

Verna, Elizabeth C. 200, 272 

Vernaz, Nathalie 1539 

Verne, Julia 564 

Vernet, Guy 1833 

Verrier, Eloi R. 1019 

Verrijken, An 928 

Verslype, Chris 171 

Verspaget, Hein W. 1238, 2069 

Verucchi, Gabriella 1802, 1844 

Vetter, Diana 1779 

Vianna, Rodrigo M. 1054 

Vibert, Eric 417, 421, 449, 487, 1237, 

1269 

Vickerman, Peter 1794, 1830 

Victor, David W. 200, 1215 

Victor, Livia B. 2072, 2076 

Vidaud, Michel 998, 1601 

Vieira, Maria Clara 2237 

Vierling, John M. 200, 624, 701, 706, 

712, 715, 1086, 1478 

Vieth, Michael 1506 

Viganò, Mauro 2011 

Vigil de Melo, Silvana 2075


Vignozzi, Linda 882, 902 

Vij, Mukul 1698 

Vijayan, Balasubramaniam 1104 

Vijayan, Santosh 183 

Vijaykumar, T. R. 284 

Vijayvergiya, Rajesh 259 

Vijgen, Leen 39 

Vila, Marta 1674 

Vila, Sergi 326 

Vilar, Graca 1859 

Vilarinho, Silvia 1719 

Vilca-Melendez, Hector 1775 

Vilchez, Regis A. 1086, 1107 

Vilchez, Valery 673, 854, 1267, 2081 

Villa, Erica 1802, 1844 

Villa, Rosa 8 

Villadsen, Gerda E. 768 

Villani, Ambra 811 

Villanueva, Augusto 254 

Villanueva, Càndid 734 

Villard, Jean 1735 

Villaret, Maxime 997 

Villela-Nogueira, Cristiane A. 273, 289, 

762 

Villeneuve, Jean-Pierre 1997 

Vilstrup, Hendrik V. 301, 768, 2071 

Vimalesvaran, Sunitha 1733 

Vinaixa, Carmen 1230, 1251 

Vincent, Catherine 609 

Vincent, Robert 160, 905 

Vinci, Maria 1802, 1844 

Vinel, Jean-Pierre 263 

Viner, Kendra 1810 

Vinikoor, Michael J. 1575 

Vinken, Mathieu 586 

Violette, Shelia 1368 

Vion, Anne-Clémence 887 

Virabhak, Suchin 1087, 1143 

Virdone, Roberto 355 

Visconti, Luca 288 

Vismara, Marta 1950 

Vispo, Eugenia 438 

Visvanathan, Kumar 996, 1225, 2262 

Viswanathan, Preeti 515, 1764, 1917 

Vitale, Alessandro 351, 356 

Vitale-Cross, Lynn 20 

Vitalis, Zsuzsanna 2090 

Vitalone, Matthew J. 341, 2123 

Vitek, Libor 1487 

Vitin, Alexander A. 878 

Vittal, Anusha 352, 1814, 1815 

Vivarelli, Marco 1751 

Viveiros, Kathleen 1592 

Vivianne, Gnemmi 2222 

Vivoli, Elisa 973 

Vlachaki, Efthimia 1126 

Vlachogiannakos, Jiannis 2012 

Vlaic, Sebastian 954 

Vogel, Arndt 344, 444, 2128 

Vogel, Robert 33, 2064 

Vogel, Wolfgang 1092, 1885 

Voidonikolas, Georgios 1479 

Voigt, Michael D. 1215 

Voitenleitner, Christian 35 

Volk, Neil 580, 1329, 2073 

Von Drygalski, Annette 1034 

von Felden, Johann 1803 

Von Roenn, Natasha 1172 

Vonderscher, Jacky 1652 

Vonghia, Luisa 797, 928 

Vongsuvanh, Roslyn 441 

Vos, Miriam B. 2232 

Voss, Jordan 565 

Voutila, Jon 118 

Vradelis, Stylianos 1900 

Vrang, Niels 978 

Vreden, Stephen G. 1846 

Vu, Vinh D. 207, 368, 386, 458, 1182, 

1561 

Vue, Jenny 1231 

Vue, Padade M. 1688 

Vukotic, Ranka 1888 

Vuppalanchi, Raj 1903, 2220 

Vutien, Philip 1668, 1805 

Vyas, Ashish 1663, 2088 

Vyas, Tanmay S. 1328, 2203 

Waasdorp Hurtado, Christine 1706, 

1708 

Wada, Fumitaka 688 

Wada, Nozomu 2236 

Wade, James 52, 1491 

Wagner, Martin 662 

Wagner, Norbert 963 

Wahl, Janice 40, 42, 210, 251, 700, 

701, 703, 712, 715 

Wahlang, Banrida 666 

Wahlin, Staffan 308 

Waintraub, Daniel J. 1095 

Wainwright, Helen 1667 

Waisbourd-Zinman, Orith 195 

Waisman, Ari 183, 1353, 1949 

Waitzberg, Dan 240 

Wakai, Toshifumi 2165 

Wakayama, Kenji 866 

Wakayama, Yuko 569, 2131, 2234 

Waked, Imam 344, 708, 733, 1163, 

1853 

Wakefield, Dorothy 1469 

Wakimoto, Yukio 1573, 1599, 1618 

Wakita, Takaji 985, 1003, 1679 

Wakui, Yuta 1648, 1943, 2167 

Walczak, Robert 974 

Walenbergh, Sofie 922 

Walewski, Jose L. 972 

Walia, Sandeep 1254 

Waljee, Akbar K. 418 

Walker, Alex J. 1140 

Walker, David R. 1050, 1139, 1152, 

1175 

Walker, Lucy J. 611 

Walker, Susan A. 1309 

Wallace, Paul K. 332 

Walsh, Christopher 1034 

Walsh, Nick 577 

Walsh, Renae 1551 

Wamhoff, Brian R. 175, 664, 915, 

1916 

Wan, Jinghong 1391 

Wan, Ying 190, 514, 817, 834, 835, 

929, 1317, 1349 

Wan, Yu-Jui Yvonne 63, 508, 649, 674 

Wan, Zhihong 1609 

Wandeler, Gilles 1575 

Wands, Jack R. 256, 1962 

Wang, Chong 2018 

Wang, Alice Y. 1087, 1143, 1144, 

1462, 2184 

Wang, Amber W. 511 

Wang, Bruce M. 683, 2111 

Wang, Cheng 1536, 1537, 2008 

Wang, Chia-Chi 1996 

Wang, Chih-Chi 1942 

Wang, Chuan 2018 

Wang, Chunping 354 

Wang, Clay 1936 

Wang, Connie W. 844, 1219, 1468 

Wang, Deli 1039 

Wang, Dongsheng 1149 

Wang, Fu-Sheng 2179 

Wang, Gary 993 

Wang, Gui-Qiang 1564, 1602, 2023 

Wang, Guo-Ying 419, 440 

Wang, Hongliang 27 

Wang, Hongtao 194 

Wang, Hongwu 909 

Wang, Hongyan 1656 

Wang, Horng-Yuan 1996 

Wang, Hua 110 

Wang, Hui 1611 

Wang, Jiang-Bin 1465 

Wang, Jianhong 754 

Wang, Jiao-Jing 54 

Wang, Jiaohong 1952 

Wang, Jiayou 879, 1324, 1338 

Wang, Jie 34 

Wang, Jing 205, 613, 1034, 1541, 

1579, 1617, 1773, 2151 

Wang, Jing-Houng 1873, 2010 

Wang, Jingyun 1580, 1620 

Wang, Jiuping 1773 

Wang, Jiyao 131, 1465, 1586, 2002 

Wang, Junfeng 74, 623 

Wang, Kasper S. 133 

Wang, Ke 1772 

Wang, Lai Mun 2190 

Wang, Lan S. 1095 

Wang, Lan Sun 2214 

Wang, Li 1336, 1865 

Wang, Li-Yu 1593, 1604 

Wang, Lin 1360 

Wang, Lirui 59, 111 

Wang, Mengjun 371 

Wang, Mo-li 1149, 1193 

Wang, Peipei 1423 

Wang, Ping 189, 1404 

Wang, Qi 1184 

Wang, Qianfan 36 

Wang, Qianyi 374 

Wang, Qiaomin 1537 

Wang, Qixia 308 

Wang, Reena 720, 728 

Wang, Renxue 824 

Wang, Rong-qi 638 

Wang, Ruisi 1315, 1355 

Wang, Shaogui 1325 

Wang, Shaoqing 1966 

Wang, Shu-Chi 2264 

Wang, Sihyung 696 

Wang, Stanley 248, 250, 719 

Wang, Tao 943 

Wang, Ting 1528 

Wang, Tingting 1537 

Wang, Weimin 510, 2116 

Wang, Wen-Jun 321 

Wang, Wenjun 468 

Wang, Xiang 1521 

Wang, Xiaofeng 1656 

Wang, Xiaohong 1865 

Wang, Xiaojie 104, 151 

Wang, Xiaojing 909, 1660


Wang, Xiaomei 1149, 1193, 1580, 

1603, 1620, 1869 

Wang, Xiaoming 374 

Wang, Xiaoxin 139, 142, 907 

Wang, Xiaoyu 183, 955, 1353, 1376 

Wang, Xintao 333 

Wang, Xue-Yan 1973 

Wang, Xuemei 130 

Wang, Xuyang 168 

Wang, Yan 193, 197 

Wang, Yang 704, 717 

Wang, Yaqi 909 

Wang, Yen-Po 738, 751 

Wang, Yifeng 140 

Wang, Yin-Chen 2050 

Wang, Ying 686 

Wang, Yongping 919 

Wang, Yongqing 22 

Wang, Yu 374, 898, 1966, 1973 

Wang, Yucai 295 

Wang, Yudong 1536, 1537, 2008 

Wang, Yuming 209, 1865, 2024 

Wang, Yunwu 328 

Wang, Zhengyu 754 

Wangensteen, Kirk J. 494, 511 

Wani, Hamid 1188 

Wanichanuwat, Jirachaya 1999 

Ward, Stephen C. 870, 1337 

Warner, Alex 1455 

Warner, Nikole L. 2142 

Washburn, W. Kenneth 1900 

Washington, Shenna 946 

Wasitthankasem, Rujipat 2036 

Wasser, Martin N.J.M. 1238 

Wat, Cynthia 245, 2026 

Watabe, Takeshi 1795 

Watanabe, Hiroshi 1912 

Watanabe, Mamoru 164, 406, 1018, 

1027 

Watanabe, Sumio 226, 794, 917, 

1771, 2158, 2193 

Watanabe, Takako 406, 1018, 1027 

Watanabe, Takao 788, 1293 

Watanabe, Takehisa 513, 1912, 1987 

Watanabe, Tsunamasa 1006, 1446, 

1655 

Watashi, Koichi 1679 

Watkins, Jennifer 2081 

Watkins, Paul B. 96, 224, 225, 228, 

1930, 1932, 1933 

Watkins, Steve 1359 

Watt, Gordon P. 1424 

Watt, Jennifer 129 

Watt, Kymberly D. 836, 1038, 1124 

Wattacheril, Julia J. 2166 

Watts, Micah M. 521 

Watts, Michelle 1172 

Waziry, Reem K. 1868 

Weatherburn, Peter 1830 

Webb, Glynn 523 

Webb, Gwilym 631 

Weber, Achim 46, 1402 

Weber, Bernd 1170, 1200 

Weber, Susan C. 359, 386, 388, 458, 

1806, 1816, 1848 

Wedd, Joel P. 2085 

Wedemeyer, Heiner 445, 1002, 1086, 

1107, 1131, 1166, 1572, 1597, 

1797, 1803, 1861, 2003, 2026 

Wee, Aileen -. 1436 

Weerachayaphorn, Jittima 661 

Weersma, Rinse K. 76 

Wehrkamp, Cody J. 60 

Wei, Lai 782, 1436, 1820, 1850, 

1877, 1973 

Wei, Lan 957, 1938 

Wei, Liang 341 

Wei, Wei 497 

Wei, Xiaoli 114, 926 

Weick, Alexander 1254 

Weigert, Roberto 20 

Weijer, Sebastiaan 1995 

Weil, Delphine 1036, 1187 

Weiland, Ola 37, 252, 1058 

Weimer, Liliana E. 1802, 1844 

Weinberg, Ethan 1823 

Weiner, Joshua 2129 

Weinger, Matthew B. 529 

Weinman, Edward J. 829 

Weinman, Steven A. 352, 1291, 1311, 

1814, 1815 

Weinmann, Arndt J. 376 

Weinreb, Paul 1368 

Weinstein, Ali A. 18, 316 

Weinstein, Jeffrey S. 1146, 1169 

Weir, Graeme 753 

Weismuller, Tobias J. 76 

Weiss, Emmanuel 265, 1288, 1391 

Weiss, Jeffrey J. 1151, 1870 

Weiss, Karl Heinz 618 

Weiss, Nicolas 1510 

Weiss, Thomas S. 954 

Wek, Ronald C. 320, 884 

Weksbergc, Rosanna 682 

Welker, Martin W. 1055 

Welle, Stephen L. 671 

Weller, Shaun 1310 

Wells, James 676 

Wells, Malcolm M. 2170 

Wells, Rebecca G. 195, 814, 1366 

Weltman, Martin 106, 627, 1343, 

1450 

Welzel, Tania M. 37, 252, 1045, 

1057, 1058, 1106, 1130, 1860, 2003 

Wen, Li 305 

Wen, Simin 2018 

Wen, Wan-Hsin 1701 

Wendon, Julia 212, 740, 1758, 1763, 

1775 

Weng, Cindy 2112 

Weng, Shih-Yen 183, 955, 1353, 1376 

Werba, Gregor 1282 

Werby, Jasper 462 

Werner, Christoph R. 1183 

Werner, Mårten 308 

West, Sara 552 

Westbrook, Rachel 764, 1216 

Westlin, William 957, 1938 

Weston, Chris J. 1296 

Wheatley, Daniel J. 1321 

Wheeler, David A. 129 

White, Duncan 536 

White, Jon 130, 682, 2118 

White, Melanie 52, 1463, 1472, 1491, 

1527 

Whitener, Melissa 1814, 1815 

Whitington, Peter F. 159, 1694 

Wiedtke, Ellen 98 

Wiegand, Steffen B. 2026 

Wiegel, Joshua 1101 

Wiesner, Russell H. 1249 

Wievel, Emily 1839 

Wigg, Alan J. 106, 627, 1077, 1264 

Wiggins, Shanna M. 136 

Wijarnpreecha, Karn 964 

Wilairatana, Polrat 555 

Wilder, Julius M. 1887 

Wildhaber, Barbara E. 1735 

Wildner, Letícia M. 2075 

Wilkey, Daniel W. 587 

Wilkins, Benjamin J. 814 

Wilkins, Kenneth J. 779, 1717, 2144 

Willacy, Oliver 337, 1388 

Willandt, Barbara 171 

Willars, Christopher 212, 1758, 1775 

Willebrords, Joost 586 

Willems, Bernard E. 1045, 1997 

Willemse, Sophie 208, 1995, 2046, 

2263 

Willemssen, Francois 1238 

Willenbring, Holger 98, 680 

Willett, Michael S. 2247 

Williams, Bianca 1314 

Williams, Michael J. 678 

Williams, Roger 163, 599, 1978, 2143 

Williamson, Kate D. 74, 622 

Williamson, Rachel M. 2168 

Willis, Sarah 1114 

Willms, Arnulf G. 43 

Willy, Jeffrey A. 320, 884 

Wilson, Antoinette 527 

Wilson, Caroline 44 

Wilson, Eleanor 92, 220, 578 

Wilson, Laura A. 107 

Wiltberger, Georg 1222 

Win, Sanda 590 

Wind-Rotolo, Megan 28 

Winkler, Anja 1927, 2108 

Winokur, Allison 2175 

Winwood, Paul J. 1413 

Wires, Emily 323 

Wise, Eric S. 1223 

Witek, James 1040 

Witt, David J. 1053 

Witters, Peter 1691 

Witthoeft, Thomas 1170, 1200 

Wöbse, Michael 1572 

Wochner, Katharina 1525 

Wockner, Leesa F. 784 

Woerns, Marcus A. 1949 

Wohl, David 1063 

Wolak, Arik 2224 

Wolfe, Lynne A. 1722 

Wolff, Rodrigo 1217 

Wolford, Dennis 725 

Wolkoff, Allan W. 321, 333, 943 

Wolter, Franziska 1287 

Won, Je Hwan 379 

Wong, Amanda A. 1110 

Wong, April 1536, 1537 

Wong, Danny 126, 1545, 1550 

Wong, David K. 243, 348, 542, 1556, 

1998 

Wong, Florence 82, 278, 570 

Wong, Grace 367, 459, 2181 

Wong, Grace LH 235, 238, 349, 1548, 

1612 

Wong, Guan Wee 312 

Wong, John B. 1592 

Wong, Kelly A. 2257


Wong, Kristin N. 388 

Wong, Linda L. 1553 

Wong, Ophelia H. 47 

Wong, Philip 1045 

Wong, Robert J. 7, 199, 201, 237, 

306, 534, 877, 1252, 1534, 1549, 

1745, 1750, 2159 

Wong, She-Yan 1261, 1276, 1452 

Wong, Tiffany C. 1746, 2000 

Wong, Vincent W. 235, 238, 349, 

367, 459, 1548, 1612, 2181 

Wong, William W. 350 

Woo, Hyun Young 749, 876, 2028, 

2038 

Woo, Jacky 2052 

Woodard, Colin 1625 

Wooddell, Christine I. 32, 2022 

Woods, Donna 71, 843, 847 

Woods, Ryan 543 

Woody, Angela 1095, 1870 

Woolbright, Benjamin 585 

Woolson, Kathy L. 523 

Woolwine-Cunningham, Yvonne J. 332 

Workowski, Kimberly 1034 

Wörns, Marcus 376 

Wrba, Friedrich 297 

Wree, Alexander 55, 969, 1371, 

1522, 1971, 2185 

Wright, Elizabeth C. 218 

Wright, Matthew C. 44 

Wright, Melanie 2147, 2208 

Wu, Bechien U. 404 

Wu, Catherine H. 223 

Wu, Cheng-Kun 1873 

Wu, Christina 1121 

Wu, Chuanghong 1814, 1815 

Wu, Chun-Ying 345, 1547, 1589 

Wu, David 223 

Wu, Dehua 1939 

Wu, Di 1656, 1683 

Wu, Diana Y. 1625 

Wu, Elizabeth 1850, 1877 

Wu, George 223, 2014 

Wu, Guihui 904 

Wu, Hao 19 

Wu, Heng 102 

Wu, Hui-Lin 2027 

Wu, Jaw-Ching 759, 1639, 1665 

Wu, Jia-Feng 1701, 1705 

Wu, Jian 366, 935 

Wu, Jiashin 879, 1324, 1338 

Wu, Jingtao 710 

Wu, Li-Chen 654 

Wu, Min 2058 

Wu, Ming-Heng 1400 

Wu, Ming-Shiang 345, 1547 

Wu, Nan 190, 782, 813, 817, 834, 

835, 929, 1317, 1349 

Wu, Pengfei 904 

Wu, Quanfeng 989, 1967 

Wu, Quanxin 2024 

Wu, Raymond 1936 

Wu, Ruihong 1149, 1193, 1603, 

1620, 1869 

Wu, Shuang 1628 

Wu, Ting 909 

Wu, Tong 185, 501, 506, 686, 1096 

Wu, Vanessa 1536, 1537, 2008 

Wu, Wen-rui 498, 1951 

Wu, Xiaoning 374 

Wu, Xing 2018 

Wu, Yan 1222 

Wu, Yang 54, 1343, 1450 

Wu, Yujing 131, 1405 

Wu, Zeguang 1660 

Wulff, Jacob 161 

Wursthorn, Karsten 1176, 1205 

Wyatt, Brooke 1151 

Wyles, David L. 248, 250, 716 

Wymer, Mark 153, 2212 

Wymore, Erin 71, 843, 847 

Wyrzykiewicz, Tadeusz 36 

Wysocki, Jacek 2043 

Xavier, Torras 734 

Xavier Brito, Leonor 150 

Xia, Jielai 468 

Xia, Yuchen 1684 

Xiang, Junxi 698 

Xiang, Xiaogang 1611 

Xiang, Xiaoyu 1947 

Xiao, Gary S. 202 

Xiao, Li 1441 

Xiao, Xiaoyan 305 

Xie, Guanhua 66, 936, 1386 

Xie, Jun 496 

Xie, Pei-yi 1755 

Xie, Qing 1586, 1611, 2002, 2013, 

2023, 2048 

Xie, Wangang 1084, 1086 

Xie, Xing-Wang 1973 

Xie, Yan 54, 911 

Xie, Yao 2048 

Xie, Yi 2048 

Xin, ShaoJie 1609, 1675 

Xing, Jian 15, 1789, 1800 

Xu, Aimin 898 

Xu, Che 1910 

Xu, Cheng 1865 

Xu, Dongping 1595, 1675 

Xu, Fujie 15, 525, 1789, 1800 

Xu, Gang 366 

Xu, Hong 54 

Xu, Hongqin 1149, 1580, 1620, 1869 

Xu, Huilei 36 

Xu, Jason J. 2083 

Xu, Jian-Ming 1465 

Xu, Jianfang 1583 

Xu, Jun 186, 187, 890, 904 

Xu, Lei-bo 498, 1951 

Xu, Ming-Jiang 110, 334, 903 

Xu, Xiaoyuan 2066 

Xu, Yang 2066 

Xu, Yongcai 67 

Xu, Yuqin 2048 

Xu, Zhihui 1595 

Xuan, Lei 548 

Xue, Bingzhong 933 

Xue, Yuhua 1979 

Yaari, Shaul 1878 

Yab, Tracy C. 169 

Yada, Akito 188, 1420 

Yada, Masayoshi 1157 

Yadav, Ajay K. 1664 

Yagi, Hiroshi 336 

Yagi, Takahito 339 

Yakushijin, Takayuki 453, 1195, 1197, 

1201, 2213 

Yalamanchili, Rachana 1185, 1202 

Yalinaycirak, Meltem 932 

Yamada, Akira 1197 

Yamada, Gotaro 2030 

Yamada, Kazutoshi 370 

Yamada, Makoto 2156 

Yamada, Masanobu 477 

Yamada, Ryoko 1195, 1197, 1201, 

2213 

Yamada, Takuya 1584 

Yamada, Yukinori 1197 

Yamagiwa, Satoshi 310, 2165 

Yamagiwa, Yoko 794, 1138, 1795 

Yamaguchi, Akihiro 181, 569, 2131, 

2140, 2234 

Yamaguchi, Kanji 886, 888, 901, 

1980, 2230 

Yamaguchi, Masakatsu 1116 

Yamai, Takuo 1661 

Yamamoto, Akiko 906, 2156 

Yamamoto, Gen 1213 

Yamamoto, Ken 1632 

Yamamoto, Kuniko 1326 

Yamamoto, Naoki 329, 1447 

Yamamoto, Takashi 1627 

Yamamoto, Yasunori 2146 

Yamanaka, Junichi 1420 

Yamane, Keiko 482 

Yamasaki, Chihiro 1679 

Yamasaki, Kazumi 408, 783, 1600 

Yamasato, Florencia 2106 

Yamashina, Shunhei 226, 917, 1771 

Yamashita, Naoki 1159 

Yamashita, Shunichi 551 

Yamashita, Taro 144, 699, 1000, 

1348, 1395, 1653, 1953, 1968, 1970 

Yamashita, Tatsuya 392, 1348, 1653 

Yamato, Masatoshi 325, 947 

Yamauchi, Akira 1937 

Yamazoe, Taiji 513 

Yan, Fengxia 790 

Yan, Hongqing 1193 

Yan, Irene K. 335, 358, 384 

Yan, Libo 1562 

Yan, Lunan 851 

Yan, Taotao 1541, 1579, 1617, 1773 

Yan, Wei 328 

Yan, Weiming 1656, 1660, 1683 

Yan, Xiaohong 239, 2153, 2154 

Yan, Zehui 1865 

Yanagawa, Takayo 1369 

Yanagi, Ami 1679 

Yanagita, Kimihiko 2031 

Yang, Zhihong 1336 

Yang, Guo-Xiang 77, 602, 613, 639, 

1285 

Yang, Heping 1952 

Yang, Hong 198, 1986 

Yang, Hushan 371 

Yang, Hwai-I 207, 1561, 1604, 1828 

Yang, Hyun 452, 1882 

Yang, Jenny C. 219, 718, 1080, 1105, 

1120, 1133, 1168 

Yang, Jenny D. 1549 

Yang, Jiaqi 361 

Yang, Jijin 851 

Yang, Jin-Hui 1465 

Yang, Jing 22, 1521 

Yang, JinMo 1615 

Yang, Jr-Shiuan 2116 

Yang, Ju Dong 127, 381, 415, 1124 

Yang, Ju-Il 1960 

Yang, Kai-ting 1945


Yang, Lei 245, 2026 

Yang, Lifei 698 

Yang, Lin 1396 

Yang, Ling 1441 

Yang, Luoluo 2135 

Yang, Man 468 

Yang, Michael 549 

Yang, Ming 1436, 1850, 1877 

Yang, Rong 709 

Yang, Ruifeng 1820 

Yang, Sera 1972 

Yang, Sheng-Shun 1996 

Yang, Sien-Sing 2050 

Yang, Su-Jau 87 

Yang, Sun 516 

Yang, Tzeng-Huey 1547 

Yang, Victor 1015 

Yang, Wenli 79 

Yang, Wucai 1536 

Yang, Xiaoyan 709 

Yang, Yang 419, 440 

Yang, Yin 38 

Yang, Ying 1306 

Yang, Ying-Ying 965 

Yang, Yongfeng 2048 

Yang, Yongping 354 

Yang, Yuan 1541, 1579, 1617, 1773 

Yang, Yuching 224, 228 

Yang, Zhen 516 

Yang, Zhiping 754 

Yanko, Adir 1032 

Yanny, Beshoy T. 1119 

Yano, Hirohisa 465, 1990 

Yao, Francis Y. 4, 50, 53, 414, 471 

Yao, Liying 1293 

Yao, Lu 185, 686 

Yao, Zhi Q. 984, 1004 

Yapali, Suna 1247 

Yaqoob, Usman 1355, 1406 

Yara, Sho-ichiro 970 

Yardeni, David 2224 

Yarmush, Martin L. 25 

Yarnykh, Vasily L. 803 

Yartel, Anthony K. 1792 

Yaseen Alsabbagh, Mohammed Eyad 

1122 

Yasuchika, Kentaro 402, 1213, 1976 

Yasuda, Katsutaro 1976 

Yasui, Kohichiroh 886, 888, 901, 

1980, 2230 

Yasui, Shin 1207 

Yasui, Yutaka 1115 

Yasukawa, Lee Ann 359, 386, 388, 

458, 1806, 1816, 1848 

Yasumura, Seiji 551 

Yasunaka, Tetsuya 2236 

Yates, Denise 260, 753 

Yates, Katherine P. 160, 2161 

Yatsuhashi, Hiroshi 408, 783, 1448, 

1600, 1632 

Yatsuzuka, Naoyoshi 707 

Yazdani, Hamza 184 

Yazdanpanah, Yazdan 1845 

Ye, Dewei 898 

Ye, Juan 366 

Ye, Wen 133 

Ye, Xin 2007, 2062 

Yeap, Sze Pheh 1264 

Yee, Brittany E. 2250 

Yee, Leland J. 241, 2014, 2025 

Yeh, Heidi 1732 

Yeh, Matthew M. 230, 923, 946 

Yeh, Ming-Lun 2264 

Yeh, Wendy W. 725, 730, 731 

Yen, Elizabeth H. 2149 

Yen, Yi-Hao 1873, 1942 

Yeo, Winnie 344 

Yeoman, Andrew D. 312 

Yeon, Jong Eun 391, 405, 1504, 1897, 

2017 

Yeung, Chun-Yan 1701 

Yi, Li-Na 1942 

Yi, Nam-Joon 400, 474, 1277 

Yi, Ruitian 1541 

Yi, Sooji 1554 

Yi, Wenjing 1004 

Yildirim, Beytullah 1613 

Yilmaz, Abdulkerim 2219 

Yilmaz, Funda 1739 

Yilmaz, Guldal 1719 

Yilmaz, Yusuf 2180 

Yim, Colina 348, 647 

Yim, Hyung Joon 391, 405, 1438, 

1504, 1897 

Yim, Sun Young 433, 1438, 1504 

Yimam, Kidist K. 76 

Yin, Xin 1703 

Yin, Chi-Biao 2023 

Yin, Hong 19 

Yin, Meng 1355 

Yin, Michael 380 

Yin, Paley 1587 

Yin, Philip 702, 706 

Yin, Wenwei 1647 

Yin, Xiaoyu 454 

Yin, Xinmin 114, 926 

Yin, Zhanxin 468, 754 

Ying, Jun 357 

Ying, Qilong 1283 

Ying, Wenbin 1433 

Yip, Benjamin 2250 

Yip, Marcus 29, 1852 

Yip, Terry C. 349 

Yoh, Kazunori 781, 1669 

Yokohama, Keisuke 966 

Yokomori, Hiroaki 1514 

Yokoo, Hideki 866 

Yokosuka, Osamu 1207, 1490, 1516, 

1628, 1632 

Yokota, Masaki 1159 

Yokote, Seiichiro 513 

Yokoyama, Keiji 385, 410 

Yokoyama, Takahide 869 

Yokoyama, Yoshinobu 1636, 1661 

Yoneda, Akihiro 1375, 1378 

Yoneda, Masato 785, 1017 

Yonem, Ozlem 2219 

Yonemoto, Koji 304 

Yong, Sherri 774 

Yoo, Byung Chul 423, 1960, 2049 

Yoo, Jeong-Ju 242, 400, 447, 470, 

474, 479, 1610, 2016, 2042, 2055 

Yoo, Suk Young 2151 

Yoo, Wonbeak 2151 

Yoo, Yang J. 405 

Yoo, Yang Jae 1897, 2017 

Yoon, Dukyong 2056 

Yoon, Jung-Hwan 242, 400, 447, 470, 

474, 479, 1610, 1907, 2016, 2042, 

2055 

Yoon, Ki Tae 876, 1542, 2028 

Yoon, Seung Kew 452, 502, 1025, 

1419, 1615, 1882 

Yoon, Yoo-Seok 857, 1902 

York, Samuel 778 

Yoshida, Atsushi 1254, 1274 

Yoshida, Eric M. 204, 1040, 1049 

Yoshida, Hitoshi 1302, 1941 

Yoshida, Kanako 1824 

Yoshida, Mariko 699, 1953 

Yoshida, Osamu 338, 788, 1293 

Yoshida, Shuhei 1379 

Yoshida, Yuichi 227, 906, 1195, 1197, 

1201, 1974, 2156, 2213 

Yoshihara, Mie 2267 

Yoshiji, Hitoshi 677, 689 

Yoshimaru, Youko 1912, 1987 

Yoshimi, Satoshi 1858 

Yoshimoto, Tsuyoshi 1159 

Yoshio, Sachiyo 476, 1643 

Yoshioka, Teppei 26, 1636, 1809 

Yoshioka, Wataru 296 

Yoshitoshi, Elena Y. 1976 

Yoshiza, Kai 1097 

Yoshizane, Yasumi 1679 

Yoshizato, Katsutoshi 191 

Yosry, Ayman 708 

Yotsuyanagi, Hiroshi 912 

Yotti, Raquel 1503 

You, Hong 374, 1404 

You, Kazunori 466, 770, 1533 

You, Min 879, 1324, 1338 

You, Qiang 1910 

You, San-Lin 1593, 1604 

You, Shaoli 1609, 1675 

Younes, Ziad 219, 1051, 1067 

Young, Elisa 106, 627 

Young, Kellie 237 

Younis, Rasha 1978, 2143 

Younossi, Issah 18 

Younossi, Zobair M. 18, 87, 153, 199, 

201, 315, 316, 534, 575, 877, 891, 

892, 956, 1035, 1079, 1177, 1186, 

1252, 1460, 1534, 1538, 1745, 

1750, 1788, 2159, 2173, 2192, 

2204, 2205, 2212 

Younoszai, Zahra 892, 2173, 2192 

Youssef, Amira S. 1821 

Yu, Amanda 86, 543 

Yu, Ami 377 

Yu, Dominic 413, 793, 1530 

Yu, Fei 709 

Yu, Ge 1149 

Yu, Haibin 1406 

Yu, Herbert 1553 

Yu, Lei 1211 

Yu, Lijia 996, 1225 

Yu, Ming-Lung 367, 398, 459, 1828, 

2264 

Yu, Su Jong 242, 400, 447, 470, 474, 

479, 1610, 1907, 2016, 2042, 2055 

Yu, Tunan 1962 

Yu, Xian-huan 498, 1951 

Yu, Yao 714 

Yu, Zhangsheng 1316 

Yu, Zu-Jiang 1616 

Yuan, Junying 832 

Yuan, Yong 711, 1456, 1461, 1828 

Yuasa, Korta 552 

Yudina, Tetyana 659


Yue, Shi 1283, 2138 

Yuen, Lilly 1551, 1557, 1563, 1566 

Yuen, Man-Fung 126, 1545, 1550, 

1746 

Yukawa, Toyokazu 2074 

Yuksel, Muhammed 305 

Yun, Tae Jung 1438 

Yurdaydin, Cihan 1572, 1678, 2004, 

2012, 2033 

Yusuf, Aasim 1872 

Zaaijer, Hans L. 1995 

Zabron, Abigail 212 

Zachou, Kalliopi 73 

Zaghla, Hassan E. 733 

Zaidan Dagli, Maria Lucia 586 

Zakharia, Kais 396, 1966 

Zamboni, Fausto 166, 1646 

Zamor, Philippe J. 17, 210, 715, 1023 

Zamora, Javier 745 

Zamora-Valdes, Daniel 836 

Zampino, Rosa 1837 

Zanetto, Alberto 351, 1270, 1889 

Zanieri, Francesca 949, 1963 

Zapata, E. 583 

Zapata, Homero A. 1281 

Zapata, Rodrigo L. 595 

Zarski, Jean-Pierre H. 1439 

Zatsiorsky, James V. 496 

Zbrzezniak, Justyna 1502 

Zehnter, Elmar 1170, 1200 

Zeilinger, Katrin 1921 

Zein, Nizar N. 840, 855, 856, 1265 

Zekri, Abdelrahman 1821 

Zemel, Michael 933 

Zenatti, Vanessa L. 2072, 2076 

Zendejas, Ivan 177 

Zeng, Changjun 685 

Zeng, Hong 498, 1951 

Zeng, Ni 1945 

Zeng, Qing-Lei 1616 

Zeng, Xianghua 1865 

Zeng, Zheng 1587 

Zeng, Zhenzhen 1634 

Zeng, Zhiping 131 

Zeremski, Marija 1841 

Zerial, Marino 617 

Zerkly, Salah 2082 

Zern, Mark 685 

Zeuzem, Stefan 37, 91, 94, 249, 252, 

445, 700, 701, 703, 714, 980, 1045, 

1055, 1057, 1058, 1086, 1106, 

1130, 1166, 1559, 1860, 1861, 

2001, 2013, 2257, 2260 

Zhai, Xiangjun 1583 

Zhan, Le 919, 934 

Zhang, Chen 1910 

Zhang, Yanqiong 2024 

Zhang, David 1405 

Zhang, David Y. 1392 

Zhang, Dazhi 1647 

Zhang, DingYu 1412 

Zhang, Dongyan 1939 

Zhang, Fang 218 

Zhang, Haiyun 1441 

Zhang, Hang 1301 

Zhang, Heng-Hui 1973 

Zhang, Hong 2058 

Zhang, Huaihong 209 

Zhang, Ji-Yuan 2179 

Zhang, Jian Q. 207, 359, 368, 388, 

1561, 1806, 1848 

Zhang, Jie 2261 

Zhang, Jingwen 1304, 1312, 1331 

Zhang, Jinjin 589 

Zhang, Jinqiang 686 

Zhang, Junlan 79 

Zhang, Kunsong 454 

Zhang, Lei 468 

Zhang, Li 904 

Zhang, Lihua 1305 

Zhang, Lumin 1397 

Zhang, Min 120, 1772 

Zhang, Ming-Xiang 2023, 2048 

Zhang, Nan 159 

Zhang, Ning-Ping 366, 935, 2002 

Zhang, Peixin 1004 

Zhang, Peng 418, 1149 

Zhang, Qin 1586 

Zhang, Qiongfang 1647 

Zhang, Renwen 2066 

Zhang, Rui 498, 1951 

Zhang, Shanshan 1948 

Zhang, Shengli 2035 

Zhang, Shulin 1579, 1617 

Zhang, Shuqin 209, 2048 

Zhang, Wei 782 

Zhang, Weici 77, 613, 619, 639 

Zhang, Wenliang 1335 

Zhang, Wujuan 916, 1695 

Zhang, Xiang 114, 926 

Zhang, Xiaogang 1773 

Zhang, Xiaoping 1660 

Zhang, Xiaxia 2066 

Zhang, Xiugen 2126 

Zhang, Yanling 685 

Zhang, Ying 984, 1004 

Zhang, Yizhou 663, 986, 1649, 1681, 

1944 

Zhang, Yu 1541, 2066 

Zhang, Yuanqing 131 

Zhang, Yuanya 1656 

Zhang, Yuguo 638 

Zhang, Yuxia 1336 

Zhang, Zheng Z. 54 

Zhang, Zhongyang 253, 254 

Zhang, Zhuoli 468 

Zhao, Changqing 359, 367, 386, 387, 

388, 428, 458, 459, 1182, 1806, 

1816, 1848 

Zhao, Chenyang 668 

Zhao, Cuiqing 1306 

Zhao, Enpeng 1333 

Zhao, Gang 828, 1924 

Zhao, Hong 1564, 1602, 1644 

Zhao, Huaying 1646 

Zhao, Jie 1311, 1814 

Zhao, Jun 1595, 1675 

Zhao, Lu-Lu 1701 

Zhao, Ming 613 

Zhao, Qiyi 120, 1423 

Zhao, Suxian 638 

Zhao, Weihan 710, 723 

Zhao, Wen Jing 209 

Zhao, Xiao 814 

Zhao, Xinyan 374 

Zhao, Yan 468 

Zhao, Yang-Jing 1973 

Zhao, Yi-Ming 366 

Zhao, Yingren 1541, 1579, 1617, 

1773 

Zhao, Yue 37, 252, 1058 

Zhao, Zhongxin 1396 

Zheng, Huanwei 2048 

Zheng, Hui 2166, 2194 

Zheng, Xinglong 698 

Zheng, Yubao 1772 

Zhong, Jin 1149 

Zhong, Li 496 

Zhong, Shuping 460 

Zhong, Wei 1335 

Zhou, Chan 778 

Zhou, Hao 675 

Zhou, Haoyue 919 

Zhou, Huiping 22, 804, 1521 

Zhou, Kali 577 

Zhou, Lili 255 

Zhou, Mei 198, 1986 

Zhou, Nannan 709 

Zhou, Tianhao 813, 1317, 1349 

Zhou, Tiaohao 929 

Zhou, Wei 2116, 2261 

Zhou, Xin-Min 1465 

Zhou, Xingliang 1283 

Zhou, Yueren 525, 1799 

Zhou, Yun 984 

Zhou, Zhanxiang 1335 

Zhou, Zhou 182, 903 

Zhu, Airu 904 

Zhu, Bao Shen 209 

Zhu, Bing 1609 

Zhu, Chuanlong 1644 

Zhu, Chuanwu 2048 

Zhu, Fusheng 2048 

Zhu, Ji-Min 1964 

Zhu, Liguo 1583 

Zhu, Lin 1660 

Zhu, Liqing 1441 

Zhu, Mansheng 498, 1951 

Zhu, Mingyue 1977 

Zhu, Qingjing 1441 

Zhu, Weiping 2048 

Zhu, Xuan 1465 

Zhu, Yanni 1041 

Zhu, Yong 120 

Zhuo, Luting 1651 

Zibbell, Jon E. 1839 

Zibert, Andree 684, 2103 

Ziegert, Szilvia 1002 

Zignego, Anna Linda 1190, 1802, 

1844 

Zimerman, Michal 679 

Zimmer, Sebastian 961 

Zimmer, Vincent 76, 463 

Zimmermann, Henning W. 828, 831 

Zimmermann, Tim 1432 

Zinski, Anne 1804 

Ziol, Marianne 132, 831 

Zipprich, Alexander 1519, 1520 

Zitsman, Jonah 2129 

Zitteli, Patricia M. 1033 

Ziv, Elad 78 

Zocco, Maria Assunta 432 

Zolfino, Teresa 309 

Zoli, Marco 355 

Zoller, Heinz M. 1092, 1885 

Zollner, Gernot 2068 

Zopf, Steffen 1506 

Zorzi, Daria 1991


Zou, Huaibin 209 

Zou, Yan 1583 

Zoubek, Miguel Eugenio 828, 1924 

Zoulim, Fabien 206, 1847, 2003 

Zozaya, Jose M. 2032 

Zuberi, Bader F. 1872 

Zubkova, Iryna 1546 

Zublena, Irène 1162 

Zuckerman, Valentina 653 

Zucman-Rossi, Jessica 132, 362 

Zürcher, Samuel 1575 

Zwahlen, Marcel 1575 

Zweers, Serge 604

1366A CATEGORY INDEX HEPATOLOGY, October, 2015 

Category Index 

All numbers refer to the corresponding abstract. 

Acute Liver Failure and 

Artificial Liver Support 

AO1. Acute Liver Failure and 

Artificial Liver Support 

Arcuate artery resistive ind 1758 

Bioluminescence imaging of m 1755 

Clinical Features and Outcom 1757 

Does Methacetin Used in Brea 1763 

Dual Role of Epidermal Growt 1767 

ELAD VTL C3A Cells May Impac 1770 

End Stage Liver Disease Pati 7 

Enhancing hepatocyte functio 8 

High CXCR-1 and CXCR-2 Expre 1762 

Indeterminate Etiology of Ac 1761 

Liver Volumetry determined b 212 

Lower fibrinogen level predi 1765 

Mesenchymal stem cell using 1760 

Nitric Oxide induced hepatic 1773 

Noradrenaline dose levels pr 1781 

Recovery of severe criticall 1769 

Replicative Stress and Cell 1764 

Single Center Experience: Hy 1784 

Acetaminophen Parent Compoun 1777 

Acute Liver Failure and ECMO 1775 

Cathepsin L-deficiency enhan 1771 

Characterization of Cerebral 214 

Clinical Features and Outcom 211 

Does weight loss surgery pre 1766 

Dual CCR2/CCR5 Antagonist Ce 1756 

Early elevation in serum IFN 1778 

Expression of Krüppel-like 1779 

Fulminant Hepatitis B Acute 1759 

Heat Stroke Leading to Acute 1768 

Hemophagocytic lymphohistioc 1783 

Histology predicts the need 1782 

Leukocyte Cell Derived Chemo 1774 

Liver specific organ failure 1780 

Safety and Tolerability of O 213 

Serial Procalcitonin Measure 1776 

The Receptor Interacting Pro 1772 

Biliary Physiology, 

Transport, Cholangiocyte 

Biology, and 

Experimental Cholestasis 

BO1. Transport, Bilirubin, 

Cholesterol, Lipids, and Bile Salts 

Bile acid induced cholestati 805 

Higher order metabolites of 808 

NF-kB is critically importan 809 

Proteomics analysis of rat c 806 

TGR5 agonists improve pancre 807 

The Ileal Bile Acid Transpor 810 

A Novel Mitochondrial Pathwa 804 

Administration of an Ileal A 19 

LKB1 regulates hepatocellula 20 

BO2. Cholangiocyte Biology 

Inhibition of Adenylyl Cycla 811 

Natural Killer T cells aggra 819 

Nlrp3 inflammasome activatio 21 

Pre-treatment with TIMP-3 pr 818 

Taurocholate Activates YAP v 22 

YAP Reduces Liver Parenchyma 813 

Cholestatic-induced biliary 812 

Glutathione depletion is cri 814 

Intraductal papillary neopla 820 

Knockout of the Secretin/Sec 817 

The bile acid Ursodeoxycholi 815 

The bile-acid receptor TGR5 816 

BO3. Experimental Cholestasis 

Anti-inflammatory and Antifi 198 

Keratin 23 represents a nove 831 

Loss of Caspase 8 in liver p 828 

RIP3-dependent signalling co 832 

Role of Cellular Inhibitor o 825 

Tetrahydroxylated Bile Acids 824 

CD39/ENTPD1 is protective in 826 

FXR-β-catenin complex and b 24 

Feedback regulation of autot 827 

Gut microbiota contributes t 833 

Hepatic CD4+ lymphocyte resp 823 

Knockdown of the melatonin r 835 

Knockdown of α7-Nicotinic R 830 

NHERF-1 assembles macromolec 829 

Prolonged usage of H1 or H2 822 

SRT1720 protects against cho 23 

The secretin receptor antago 834 

Wnt/β-catenin cooperates wi 821 

Cell and Molecular 

Biology 

CO1. Cell Structure and Function 

Acute alcohol binge causes s 334 

CD45+ fraction in murine adi 325 

Cataplerosis of α ketogluta 330 

Cross-talk between autophagy 326 

Genetic Induction of Metabol 331 

Hepatocyte Isolation from Li 332 

Histone 3 lysine 9 trimethyl 327 

Interaction with collagen ma 329 

Small Anti-Warburg Molecules 319 

UPR Regulation of Autophagy 320 

Annexin A6 is necessary for 64 

Characterization of Microtub 333 

Hepatocyte surface localizat 321 

Longitudinal monitoring of h 323 

Loss of adherens junctions p 255 

Mouse CD11b+Kupffer cells re 324 

Netrin-1 Promotes Liver Canc 328 

Tissue-Specific Subcellular 322 

CO2. Signal Transduction and 

Nuclear Receptors 

Akt-mediated FoxO1 inhibitio 65 

Aspartate β-hydroxylase mod 256 

Cerium oxide nanoparticle tr 659 

Cyclic AMP increases canalic 661 

Effects of Targeting Wnt/β- 673 

FGF21 facilitates normal liv 649 

HMGB1 interacts with AIM2 to 656 

Hedgehog regulates Yes-assoc 66 

Hepatic XBP1 deletion promot 257 

Hyperammonemia activates a l 668 

Hyperammonemia impairs skele 671 

IRAKM-Mincle axis contribute 675 

Lysophosphatidic acid recept 653 

MYCN is A Novel Biomarker fo 657 

Metabolomics-Targeted Pertur 672 

Orchestration of BH3-only pr 669 

RA and butyrate induce liver 674 

Regulation of the Hepatic Dr 655 

Restoration of Hepatic Physi 664 

Skeletal muscle hyperammonem 667 

Skeletal muscle mitochondria 651 

Thr505 RelA phosphorylation 652 

Circulating extracellular ve 109 

Effects of Ursodeoxycholic-a 662 

Farnesoid X Receptor (FXR) A 658 

HNF4α reprogramming during 650 

Hepatitis B virus X protein 654 

Involvement of Endoplasmic R 663 

NK cells from homozygous NLG 660 

Organopollutant Exposures Fu 666 

Sustained Inhibition of Live 670 

The regulation of liver volu 665 

CO3. Gene Expression and Therapy 

A proof of concept nonclinic 505 

Chenodeoxycholic acid induce 520 

Chimeric hepatitis E virus-l 502 

Defective mTOR-TFEB signalin 501 

Dynamic shift of microbiota 63 

Evaluation of Glyoxylate and 510 

Identification of FGF2 as a 517 

Impaired skeletal muscle mit 518 

Is Disulfiram a Potential Th 516 

MicroRNA-192 regulates the e 519 

Molecular heterogeneity in m 253 

Organic Cation Transporter-1 506 

Shifting gut microbiota and 508 

Stabilin-mediated endocytosi 507 

The Roles of Kras Isoforms i 504 

Therapeutic Potential in Tox 515 

YAP Promotes Cyclin D1 Expre 514 

Cell-Type-Specific Expressio 511 

Disruption of the HNF1α bin 492 

High-throughput sequencing t 503 

Identifying novel therapeuti 500 

In vivo distribution of exos 512 

Integrated proteomics analys 513 

Methylation-associated silen 498 

MicroRNA-122 (miR-122) Regul 493 

Mixed hepatocellular-cholang 254 

Molecular mechanisms underly 499 

Niclosamide ethanolamine inh 497 

Overexpression Screening to 494

HEPATOLOGY, VOLUME 62, NUMBER 1 (SUPPL) CATEGORY INDEX 1367A 

Peretinoin, an Acyclic Retin 495 

Pre-operative plasma glypica 509 

Therapeutic overexpression o 496 

CO4. Stem Cell Biology 

CD34+ Liver Cancer Stem Cell 685 

Decellularized Spleen Matrix 698 

Detection of putative cancer 695 

Disruption of TGF-β-regulat 682 

Ex vivo-expansion of human C 688 

Expression pattern of the st 687 

FAP disease modelling using 684 

Interleukin-17 mediates live 693 

Niche specific insulin signa 691 

Sonic Hedgehog-Containing Ex 692 

TGF-β inhibits lncRNA H19 e 686 

Wnt/beta-catenin signaling a 699 

Attenuation of liver fibrosi 689 

Bone Marrow Mesenchymal Stem 690 

Clonal analysis of the human 101 

DNMT1 is essential for effec 677 

Growth Factor Mobilized CD34 694 

Hepatic maturation of induce 676 

Hepatocyte plasticity underl 680 

Human Induced Pluripotent St 258 

Identification of hepatic st 681 

Identification of mechanism 697 

Microbial-Derived Cues Drive 679 

Notch and Wnt control ductul 678 

Self-renewing diploid Axin2+ 683 

TSG-6 enhances autophagy in 696 

Health Services Research 

DO1. Health Care Delivery/Access/ 

Quality 

Charges for Patients Hospita 533 

Communication, sequence and 529 

Development of a Model to Pr 14 

Did improvement of access to 524 

Diminished Health-Related Qu 522 

Evacuation after the Fukushi 551 

Hepatitis C Disease Burden i 104 

In Patients with Non-alcohol 575 

Learning the Liver: The Pilo 544 

National Estimates of Health 560 

Novel Algorithm to Predict H 580 

Outcome Indicators In Primar 571 

Outreach invitations improve 548 

Predicting survival in liver 555 

Rates of liver fibrosis asse 561 

Redesigning Outpatient Care 538 

Reduced Work Productivity (W 18 

Reducing Wait Times for Radi 542 

Successful management of pre 567 

The Prevalence and Mortality 534 

Thirty-day Readmission Follo 531 

Thirty-day Readmission for I 550 

Timing of Hepatitis B and C 543 

Wide variations in Albumin U 570 

Worse Outcomes Among Uninsur 563 

A simple educational 13 

A New Approach to Patient Ce 535 

A Prospective Study of a Pro 17 

A Regional Multidisciplinary 521 

A qualitative assessment of 547 

Abdominal Ultrasonography Gu 554 

Age, race and insurance stat 539 

Characteristics of Incident 562 

Comparable hepatitis C treat 541 

Deleterious effects of cirrh 566 

Depression and Alcohol Misus 525 

Disparities in the Quality o 553 

Distance to Transplant Cente 565 

Effect of HCV treatment outc 15 

Engagement in care of high r 549 

Epidemiology and survival in 532 

Epidemiology, treatment-patt 559 

Evaluation of DAA (direct-ac 527 

External Validity of Trials 574 

Geriatrics and cirrhosis: ch 528 

Heat Mapping Incident PBC Pa 546 

Hepatitis B Testing Prior to 540 

Hepatitis C Virus Sustained 573 

Hepatitis C care and treatme 568 

High Hepatitis E seroprevale 523 

High Rates of Hepatitis C Vi 545 

Interventions to optimize re 577 

Knowledge about hepatitis B 572 

Living Liver Donors: Dispari 556 

Modernizing the Child-Turcot 16 

Palliative Care Services are 557 

Physician versus Patient Per 537 

Primary Care Physician Persp 576 

QI in Practice: Reducing Rea 536 

Reduction of 30 day Readmiss 530 

Screening for poor prognosis 564 

Suboptimal Quality Of Preven 552 

Successful Implementation of 579 

The Burden of Alcohol Liver 526 

The loss of skeletal muscle 569 

Upper Limit Normal of Alanin 578 

“Normal” Serum ALT/AST L 558 

DO2. Cost-Effectiveness and 

Economics of Care 

Cost-effectiveness of Daclat 1461 

Hospital Based Initiative to 1452 

Superiority and Durability o 1460 

The Attention-to-Burden Inde 1451 

The Cost of Making Hepatitis 151 

Treating Hepatitis C in the 152 

A Trial-based Model of Liver 155 

A probabilistic decision mod 1455 

Bariatric surgery for the tr 154 

Cost-effectiveness of Sofosb 1453 

Hospitalisation for primary 1457 

Impact of Comorbidity on Inp 1459 

Impact of Daclatasvir-Sofosb 1456 

Living Donor Liver Transplan 1454 

Obeticholic Acid (OCA) has I 1458 

The cost-effectiveness of om 1462 

DO3. Health Economics and Cost 

Effectiveness of Viral Hepatitis 

Cost-effectiveness analysis 1536 

Cost-effectiveness analysis 1537 

Costs per Diagnosed Liver Di 156 

Hepatitis C (HCV) Infection 153 

Inappropriate Laboratory Uti 1535 

Rethinking pricing policy of 1539 

The Value of Cure Associated 1538 

All-oral direct acting antiv 1534 

Hepatitis B 

EO1. Virology, Pathogenesis, and 

Immunology 

An impact of cellular core-f 1642 

CRISPR/Cas9 “double”-nickase 1629 

Comprehensive analysis of th 1648 

Defective humoral immunity i 1663 

Deletion of APOBEC3B is Asso 1651 

Differential Serum Cytokine 1668 

Endogenous Antigen Presentat 1640 

Factors associated with decr 1677 

GORASP2 regulates HCV and HB 1630 

Genetic association study fa 1681 

Genome wide miRNA: mRNA Inte 1659 

Genome-wide scan of miRNA po 1649 

HBcrAg during nucleos(t)ide 1653 

Hepatitis B Virus Surface Pr 1656 

Humic matter inhibits HBV-in 1664 

IFN-α-mediated Base Excisio 1684 

Initial sites of hepadnaviru 1641 

Large Hepatitis Delta Antige 1665 

Long-term follow-up study of 1674 

Massive Intrahepatic Product 1646 

Modeling Hepatits B Virus in 1680 

Noninvasive markers of liver 1676 

Osteopontin is involved in c 163 

Precore and Basal Core Promo 1667 

Presence of naïve-like CD8+ 1666 

Proline142 deletion in the H 1639 

The mechanism of interferon 1644 

The Combination of KIR and H 1633 

The HBx-DLEU2 lncRNA complex 165 

The HLA-DPA1 rs3077 TT polym 1638 

The expansion of CD11b-CD27- 1647 

Viral Expression and Molecul 166 

A Novel Entecavir Resistance 1655 

Analysis of the effect on HB 1627 

Characterisation of the immu 167 

Comparison of detection rate 1675 

Difference of intracellular 1645 

Downregulation of TLR2 on CD 1683 

Dynamic Analysis Of Cellular 1672 

Effect of a novel synthetic 1652 

Effects of HLA-DPB1 genotype 1632 

Free episomal and integrated 1654 

Functional restoration of CD 1660 

Generation of human peripher 1635 

Genetic Variation in Vitamin 1631 

Geranylgeranylacetone exerts 1671 

Hepatitis B virus X protein 168 

Hepatitis B virus down regul 1685 

Hepatitis delta virus-induce 1650 

Human cytotoxic T lymphocyte 1670 

Human induced pluripotent st 164 

IFN-λ3 induction by nucleot 1673 

Impact of toll-like receptor 1658 

Indoleamine-2, 3-dioxygenase 1643 

Inhibition of epigenetic reg 1628 

Knockdown of NTCP reduces su 1661 

NFATc3 exerts anti-viral and 1634 

NK cell function is restored 1626 

NK cells play important role 1636


No Detectable Resistance to 1678 

Novel robust in vitro hepati 1679 

Possible involvement of hepa 1669 

Serial Changes of Th1 and Th 1682 

The Association of vitamin D 1662 

The activating receptor NKP4 1657 

The role of HBV quasispecies 1686 

Unique Liver Transcriptomic 1637 

EO2. Diagnostics, Epidemiology, 

Prevention, and Natural History 

Aldehyde dehydrogenases-2 rs 1580 

Application of Highly Sensit 1573 

Assessing the impact of hepa 1546 

Baseline quantitative hepati 1604 

Clinical Features of Chronic 1587 

Clinical outcome of long-ter 1581 

Clinical scoring system for 1616 

Comparative Study for Anti-H 1599 

Conconrdance between transie 1621 

Different DNA methylation pa 1553 

Evaluation of HBV infection 1622 

Four years real-life experie 1544 

Genotype-Specific Prevalence 1559 

HBV-related cirrhosis in the 1578 

Hepatitis B Core-Related Ant 1585 

Hepatitis B Core-Related Ant 1586 

Hepatitis B virus infection 1543 

Hepcidin: A new clinical fac 1620 

High Rates of Surface Antige 1549 

Individualizing Hepatitis B 1560 

Liver fibrosis progression, 1548 

Long term clinical outcome i 1558 

Lower Incidence of Hepatocel 1561 

Mapping HBsAg epitope profil 1551 

Nationwide Prospective Surve 1577 

Objective Determination of H 121 

PreS mutations analyzed by d 1565 

Prevalence and Molecular Vir 1566 

Prevalence and serological f 1555 

Prevalence of Hepatitis B vi 1596 

Relationship between hepatoc 126 

Risk assessment of hepatocel 1605 

Seroprevalence and Clinical 1598 

Targetting HNF4α via Activa 1562 

The Predictive Values of Hep 246 

The Usefulness of CLIF-SOFA 1609 

The role of hepatitis B core 1550 

Utility of quantitative hepa 1576 

Angiopoetin-like protein 2 i 1564 

Anti-platelet therapy reduce 1610 

Assessment Of Treatment Resp 1624 

Assessment of liver steatosi 1582 

Barriers to obtaining approp 1608 

Baseline Hepatitis B core re 1572 

Biomarkers of Fibrosis in Lo 1607 

Changes in hepatitis B virus 1584 

Changes of Hepatitis B surfa 1615 

Characterization of Nucleic 1603 

Clinical Outcomes of Patient 1556 

Collaborative Public Health 1574 

Comparing antiviral efficacy 1542 

Comparison between spontaneo 1552 

Detection of genetic and epi 1554 

Does Cesarean section and fo 1619 

Elevated serum levels of WFA 1600 

End-of-therapy Serum Gradien 1547 

Evaluation of Chronic Hepati 1606 

Evaluation of Serum Cell Dea 1611 

Factors Associated with the 1545 

HBsAg transferring from moth 1579 

Health care disparity in del 1569 

Hepatic B flares in rheumato 1589 

Hepatitis B viral load in dr 1575 

Hepatocellular Carcinoma (HC 125 

Hepatocellular Carcinoma Inc 1540 

High-throughput and ultrasen 1614 

Identification of a non-inva 1601 

Inadequate Treatment of Chro 1625 

Increased Viral Diversity is 1557 

Influence of the ethnic stat 1613 

Interleukin-2 Receptor and T 1602 

Is booster beneficial after 1623 

Is there any rule for antivi 1541 

Kinetics of Serum Hepatitis 1567 

Length of antiviral therapy 122 

Longitudinal analysis of com 1595 

Performance Assessment of Co 1618 

Performance of the new cobas 1597 

Postpartum hepatic flare aft 1617 

Prediction of Clinical Outco 1591 

Prevention of perinatal tran 1583 

Quantitative Hepatitis B sur 1571 

Redefining the Natural Histo 1563 

Serum Alanine Aminotransfera 123 

Serum HBV RNA is an early ma 1594 

Serum Quantitative Levels of 1593 

Statin and the risk of hepat 1612 

The PAGE-B Score Accurately 1590 

The PAGE-B Score Stratifies 1568 

The Role of Surface Antibody 1592 

Ultrasound predicts steatosi 1570 

What is predicting post-preg 1588 

EO3. Therapeutics: New and 

Approved 

“Real Life” outcomes for Ten 2047 

A Multicenter, Prospective, 2048 

A Randomized Prospective Ope 1995 

A randomized study of standa 1997 

A study to evaluate the dyna 2028 

Association between Pre-S2 D 2040 

Association between ALT flar 2059 

Association of Interferon-ga 2036 

Baseline HBsAg titer allows 2045 

Biochemical, Virologic, and 243 

Biologic Responses to GS-477 2052 

Characterization of Hepatiti 2026 

Comparative Effectiveness of 244 

Comparison of the efficacy o 2055 

Decreasing risk of hepatocel 2012 

Discontinuation of Antiviral 2049 

Discontinuation of Effective 2019 

Dynamics of hepatitis b viru 2066 

Early Decline in Serum HBsAg 2014 

Early prediction of response 2001 

Efficacy and Safety of Pegin 2023 

Encapsidation and secretion 2064 

Final Results of Peginterfer 2002 

GalNAc-siRNA conjugate ALN-H 36 

Inhibition of Hepatitis B Vi 33 

Intrahepatic IP-10 expressio 2046 

Liver Gene Expression Profil 2027 

Long Term Efficacy and Safet 2004 

Long term follow up of chron 2017 

Longitudinal PAGE-B Scores P 2003 

Monthly dosing of ARC-520 in 2022 

Nucleos(t)ide Analogue Disco 1998 

Nucleos(t)ide Analogues agai 242 

Plasma MicroRNA Levels are A 2005 

Prolongation of interferon t 2033 

Reduction of HBsAg by peg-in 2029 

Reductions in cccDNA under N 32 

Safety and Efficacy of GS-47 2015 

Serologic and Virologic Chan 2043 

Serum HBV RNA is an Early Pr 245 

Serum hepatitis B core-relat 2063 

Tenofovir Disoproxil Fumarat 209 

The New Cyclophilin Inhibito 2035 

The baseline combination of 2044 

The incidence and predictors 2010 

The relationship between pat 2057 

Treatment and HBeAg-status D 241 

Urinary β2-microglobulin fo 1999 

Week 24 HBsAg levels predict 2013 

Whole-Genome Sequencing of P 2037 

A randomized trial evaluatin 2038 

Antiviral Activity of Tenofo 2021 

Antiviral treatment for chro 2060 

Bone toxicity during long-te 2053 

Characterization of HBsAg lo 2032 

Combined hepatitis B core-re 2020 

Descriptive Comparison of Ef 2025 

Development and Recurrence o 2065 

Development of a Direct RNA 2062 

Dual-gRNAs and gRNA-microRNA 34 

Entecavir and Tenofovir More 2016 

Excellent outcomes of hepati 2000 

Functional Activation of NK 2009 

HBV Does Not Modulate the Tr 35 

Impact of pre-treatment plat 2031 

Is efficacy of tenofovir in 2042 

Long-Term Clinical And Serol 2008 

Long-term Renal Safety of Ad 2056 

NGAL can act as a renal safe 2054 

Prediction of sustained off- 2050 

Reduction of hepatocellular 1996 

Safety, antiviral activity, 2058 

Sequential therapy with Peg- 2030 

Study on post-treatment rela 2061 

TKM-HBV, a Novel RNA Interfe 2007 

TKM-HBV, a Novel RNA Interfe 2034 

Tenofovir-associated Fanconi 2006 

The Efficacy And Safety Of T 2024 

The Relationship Between Tel 2051 

The combination of IFN-Lambd 2011 

The impact of timely initial 2018 

Update on the safety and eff 31 

Hepatitis C 

FO1. Virology, Pathogenesis, and 

Immunology 

A methyl donor, S-adenosylme 1022 

Alcohol increases the produc 30 

Altered frequencies and func 1007 

Analysis of genetic variatio 1028 

Daclatasvir and Sofosbuvir i 217 

Different HCV genotypes have 999 

Elevation of Galectin-9 is a 26 

Elucidating Mechanisms Under 1017 

Evaluatiin of protective and 992 

Evolution Of Resistance Asso 220 

Expression of IFNλ4 in live 1018


Fast and deep early HCV-RNA 222 

HCV RNA and HCV core antigen 1002 

HIV and HCV co-infection in 25 

Individualized DAA treatment 982 

Interferon lambda 3 (IL28B) 980 

Iron chelation restores mito 995 

Lipid-Free apolipoprotein E 1019 

Liver pDCs from HCV-Infected 988 

Long noncoding RNAs regulate 991 

MicroRNA 17 Host Gene Protei 1011 

Persistence of HCV-specific 1031 

Polymorphisms in irisin-prod 221 

Pre-treatment levels of miR- 1023 

Rapid normalization of antiv 28 

Role of very-low-density lip 997 

Selenoprotein P regulates th 990 

Serial change of resistant a 1027 

The Antibody response to hep 1033 

The Emergence of NS5B Resist 219 

The impact of HLA-DQs and IF 986 

The roles of unphosphorylate 1025 

The tumor suppressor IQGAP2 1015 

Transcriptomic analysis of C 1029 

Type III interferon response 1006 

Variability of the hepatitis 1021 

Very low prevalence of Hepat 1032 

De novo formation and matura 27 

A miR-122/miR-224-based mode 998 

Activation of Hepatic Stella 29 

Analysis of AL-516, a Novel 993 

Cell-specific peripheral inn 996 

Differential microRNA expres 1014 

Downregulation of let-7 fami 1005 

Effects of Resistance Mutati 985 

Essential Factors for Recons 1003 

Establishment and Characteri 1000 

Establishment of A Novel Hep 989 

Hepatitis C Genotype 4r Resi 1009 

Hepatitis C virus facilitate 984 

Interferon-based treatment a 1008 

Kidney involvement in HCV Ch 983 

LECT2 specifically induced b 1010 

MicroRNA-130a inhibit HCV pr 981 

Novel effect of NS5A inhibit 1013 

PD-1 blockade enhances cytop 1001 

Rapid Decline In Interferon 218 

Regulation of Hepatitis C Vi 994 

Single-strand RNA-induced mo 1016 

Substitution in amino acid 7 1020 

The FGL2:FcγRIIB Immunosupp 1012 

The Hepatitis C Virus modula 987 

The effect of sofosbuvir-bas 1030 

The expression of Immune-miR 1026 

Tim-3 Inhibits Monocyte Func 1004 

FO2. Diagnostics, Epidemiology, and 

Natural History 

MICA SNP, but not DEPDC5, PN 1824 

Hepatitis E in haematologica 1803 

A Highly Specific and Sensit 85 

A New Simple Quantitative Li 1849 

Acute HCV recurrence after l 1889 

Acute hepatitis and reactiva 1882 

Age and Risk Factor-based Se 1823 

Application of a Novel Hepat 1785 

Benefits of HCV eradication 1808 

Can HCV Antigen Testing Repl 1826 

Cannabinoid receptor 2 (CB2) 1837 

Cascade of Care for Hepatiti 1786 

Cascade of Care of HCV & HIV 1891 

Comparison of performance ch 1817 

Correlates of Successful HCV 1892 

Differential Progression to 1848 

Dynamic change of α-fetopro 1873 

Effectiveness and cost-effec 1845 

Evaluation of liver and sple 1862 

Evaluation of liver and sple 1879 

Evaluation of the role of he 1890 

Expanded eligibility to HCV 1861 

Fibrosis Regression After He 1876 

Healthcare Utilization and C 1883 

Hepatitis C Seropositivity i 1822 

Hepatitis C Virus (HCV) Mort 86 

Hepatitis C Virus (HCV) Prev 1859 

Hepatitis C Virus (HCV) Prev 1818 

Hepatitis C and Renal Diseas 1867 

Hepatocellular carcinoma (HC 1847 

Highly accurate HCV genotypi 1812 

Illegal drug use disclosure 1857 

Incidence and Predictors of 90 

Increasing Prevalence of Cir 88 

Investigation into the high 1865 

Linkage to Care and Treatmen 1874 

Low HCV Treatment for Chroni 1816 

Male gender, genotype 3, pre 1844 

Mathematical model for early 1821 

Modeling the probability of 1854 

Non-invasive assessment of l 1834 

Performance of HCV Ag quanti 1833 

Policy Implications of Dispr 1813 

Pragmatic Non-invasive test 1856 

Prevalence and Risk-Factors 1853 

Prevalence of diabetes and a 1877 

Prevalence of hepatitis B co 1850 

Prevalence of viral hepatiti 1796 

Progression to liver cirrhos 1806 

Rate and Predictors of Serum 1881 

Repeated biopsies within 23 1878 

Risk of hepatitis C virus in 1793 

Safety and Efficacy of Inter 1885 

Screening Urban Baby Boomers 1835 

Self-reported Alcohol Use at 1880 

Stability and prevalence of 1832 

The Utility and Limitations 1799 

The hepatitis C epidemic amo 1830 

The influence of Hepatitis C 1869 

The modeled impact of improv 1842 

The potential prevention imp 1794 

Times they are a changing: H 1871 

Treatment prioritization acc 1802 

Trends in end-stage liver di 1868 

Wisteria floribunda agglutin 1851 

Ability of EMR-based fibrosi 1791 

Access To Reimbursement Of I 1886 

An Integrated Linkage to Car 1810 

Benefits of Antiviral Treatm 1828 

Can Hepatitis C be eliminate 1872 

Cause of death in HCV patien 1789 

Characteristics of Patients 1792 

Comparative analysis of onli 1827 

Comparison of On-Treatment H 1860 

Development of End Stage Liv 1790 

Distribution of pre-existing 1795 

Does pregnancy protect again 1884 

Evaluation of both Fucosylat 1809 

Factors Associated with Hepa 1839 

Fibrosis Progression in Pati 1841 

From Care to a Cure: Improvi 1801 

HCV in Semen of HIV-infected 1852 

Hepatitis C Birth-Cohort Tes 1787 

Hepatitis C Eradication with 1866 

Hepatitis C RNA assay differ 1797 

Hepatitis C Virus (HCV) Erad 1825 

Hepatitis C Virus (HCV) Geno 1798 

Hepatitis C Virus Genotype A 1820 

Hepatitis C prevalence, dete 1846 

Heterogeneous IL28B/IFNL4 di 1855 

Hyperbilirubinemia in HIV-HC 1840 

Impact of Hepatitis C (HCV) 1807 

Impact of comorbidities on t 1863 

Improvements in the Hepatiti 1870 

Increasing Hepatitis C Virus 1811 

Linkage to Care Outcomes for 1804 

Liver fibrosis stage and com 1800 

Natural History of Decompens 1815 

Non-Invasive Fibrosis Scores 1843 

Patients with Hepatitis C (H 1814 

Perceived risk and HCV knowl 1864 

Positive Impact of Point of 1831 

Predictive value of APRI and 1887 

Racial Differences in Socioe 1805 

Rapid, sensitive, and accura 1858 

Rectal Shedding of HCV in HC 89 

Regression of Advanced Fibro 108 

Serum levels of Wisteria flo 1838 

The Association of Sustained 87 

The Power of the Law: Improv 1875 

The clinical profiles of Jap 1819 

The epidemiology of hepatiti 1829 

The use of hepatitis C core 1888 

Treatment With Statins Reduc 1788 

FO3. Therapeutics: Preclinical and 

Early Clinical Development 

A Single Dose of RG-101, a G 208 

Miravirsen Dosing in Chronic 2255 

Pharmacokinetics, Safety and 2260 

Pharmacokinetics, Safety, an 2259 

SB 9200 Shows Antiviral Acti 2262 

Safety, Efficacy and Tolerab 2256 

Short-term safety and pharma 2261 

The effect of microRNA let-7 2264 

Treatment with the Anti-miRN 2263 

Discovery of AT-337 and AT-3 2266 

Drug-Drug Interaction Studie 2265 

EDP-239, a novel HCV NS5A in 2257 

The impact of α-fetoprotein 2267 

Toxicology evaluation of TT- 2258 

FO4. Therapeutics: New Agents (not 

approved, phase 2-3) 

98%–100% SVR4 in HCV Genot 41 

A Randomized Controlled Tria 205 

Absence of significant drug- 723 

An Integrated Analysis of 40 42 

An integrated safety analysi 716 

Analysis of HCV Genotype 1 V 705 

Baseline HCV NS5A resistance 709 

Characterization of HCV Resi 718 

Daclatasvir exposure alone d 728 

Daclatasvir exposure does no 720 

Daclatasvir in combination w 252 

Daclatasvir plus sofosbuvir 206 

Drug-drug interactions betwe 710 

Efficacy and Safety of Co-Fo 708 

Efficacy and Safety of Ombit 714


Efficacy of Grazoprevir (GZR 715 

High Efficacy of Grazoprevir 251 

High Efficacy of Grazoprevir 210 

High Efficacy of the Combina 701 

High SVR4 Rates Achieved Wit 250 

High SVR4 Rates Achieved Wit 248 

Improvement in liver disease 706 

Integrated Safety Analysis o 726 

No Clinically Meaningful Pha 725 

No Pharmacokinetic Interacti 731 

No Pharmacokinetic Interacti 730 

Ombitasvir/Paritaprevir/r an 722 

Predictors of Response to Gr 700 

Resistance Analysis of Treat 713 

SVR12 results from the Phase 39 

Safety and Tolerability of G 712 

Safety and efficacy of dacla 37 

Short-duration therapy with 702 

Sofosbuvir/GS-5816 Fixed Dos 249 

The Combination of Grazoprev 703 

A Pilot Evaluation of Twice 724 

C-EDGE CO-STAR: Efficacy of 40 

C-EDGE Co-Infection: Impact 729 

C-EDGE TN: Impact Of 12-Week 717 

Comparative Efficacy and Tol 711 

Efficacy, Safety And Pharmac 707 

High Rates of SVR in Treatme 247 

Pharmacokinetics of Coadmini 719 

Pharmacokinetics of Narlapre 721 

Prevention of allograft HCV 704 

Projected Long-Term Impact o 727 

Sofosbuvir/GS-5816+GS-9857 f 38 

FO5. Therapeutics: Approved Agents 

(THE SOFGER TRIAL) Sofosbuvi 1078 

IFNL4 ss469415590 Polymorphi 1149 

Adherence to All-Oral HCV Tr 1063 

Asian Patients with Chronic 1080 

Asian Patients with Genotype 1105 

Association of polymorphisms 1055 

Clinical Management of Ribav 1067 

Co-morbidities and co-medica 1140 

Combination of IFNL4 polymor 1153 

Combination therapy with dac 1125 

Comparison of 8 vs. 12 weeks 1036 

Demographic Predictors of Di 1048 

Early and persistent increas 1070 

Early dose adjustment of RBV 1162 

Effect of Chronic Kidney Dis 1088 

Effectiveness of Ledipasvir/ 93 

Effectiveness of Treatment o 1155 

Efficacy and Adverse Event P 1119 

Efficacy and Safety of Simep 1118 

Efficacy and Safety of Sofos 1165 

Efficacy and safety of IFN-f 1092 

Efficacy of Ledipasvir plus 1146 

Evaluation of the efficacy a 1191 

Global prevalence of pre-exi 1044 

Hepatic Decompensation and S 1185 

Hepatitis C treatment with S 1102 

Hepatitis C virus NS5A L31V 1082 

Hepatocellular carcinoma dev 1056 

High efficacy of a 12-week s 1179 

High efficacy of ledipasvir/ 1049 

Highly Successful Retreatmen 92 

IFN-λ inhibits miR-122 tran 1096 

IFN-free DAA regimens improv 1204 

Impact of baseline albumin l 1040 

Lower 25-OH Vitamin D Levels 1109 

NS3 Q80K Polymorphism in Vir 1127 


Outcomes of ribavirin free r 1174 

Patient Decision-Making: The 1114 

Patient Expectations of the 1152 

Patient Reasons for Initiati 1139 

Pharmacokinetic Analyses of 1133 

Pre-Existing Co-Morbidities 1200 

Prediction for poor virologi 1062 

Prevalence of Pre-Treatment 91 

Quality-Adjusted Cost of Car 1079 

Quantifying and Predicting E 1145 

Real Life Treatment Outcomes 1154 

Real-world effectiveness and 1176 

Real-world effectiveness of 1205 

Resistance Analyses of Phase 1168 

Retreatment with sofosbuvir 1123 

Ribavirin levels at treatmen 1098 

Safety and Efficacy of All-O 1126 

Safety and Efficacy of Sofos 1057 

Safety and Efficacy of Treat 1128 

Simeprevir plus sofosbuvir f 1072 

Simeprevir, pegylated-interf 1192 

Sofosbuvir, Ledipasvir in IB 1075 

Sofosbuvir (SOF) and Ledipas 1035 

Sofosbuvir + Ledispasvir Com 1101 

Sofosbuvir Plus Ribavirin Wi 1094 

Sofosbuvir and Ledipasvir/So 1120 

Sofosbuvir and Ribavirin for 1083 

Sofosbuvir and ledipasvir fo 1081 

Sofosbuvir in combination wi 1091 

Sofosbuvir plus Ribavirin in 1076 

Sofosbuvir-based antiviral t 1158 

Sofosbuvir-based treatments 1156 

Sofosbuvir-based treatments 1060 

Sustained Virologic Response 1043 

The TOSCAR Study: Sofosbuvir 1077 

The effect of interferon-fre 1047 

Treatment Outcomes With 8, 1 94 

Treatment of Chronic HCV Gen 1170 

Treatment of Chronic Hepatit 1122 

Treatment of Hepatitis C Gen 1163 

Treatment of Hepatitis C Vir 1186 

Virological and Clinical Res 1190 

What does Fibroscan® measur 1137 

Is ribavirin actuall 1187 

ITPA polymorphisms are predi 1193 

Interferon Signaling and the 1132 

A Meta-Analysis of the Assoc 1129 

A comparison of direct seque 1061 

Adherence and Discontinuatio 1050 

An Actuarial View of the Agi 1069 

Approved All-oral Sofosbuvir 1034 

Assessment of serum hepatiti 1207 

Association between severe r 1138 

Awareness Of Hepatitis C Sta 1042 

Better Work Productivity and 1198 

Cardiac arrhythmia in patien 1194 

Clinical Significance of Blo 1203 

Clinical- and Economic-perfo 1095 

Combination therapies with d 1150 

Concomitant use of chemother 1160 

Cost-effectiveness of ombita 1143 

Daclatasvir plus sofosbuvir 1058 

Detecting Drug-Induced Liver 96 

Determination of on-treatmen 1103 

Differences in NK phenotype 1117 

Early response and efficacy 1167 

Effect of Different Immunosu 1045 

Effect of Proton pump inhibi 1199 

Effect of Renal Function on 1066 

Effectiveness of 12 or 24 we 1108 

Effectiveness of 8 or 12 wee 1046 

Efficacy and safety of dacla 1116 

Efficacy and safety of ombit 1107 

Efficacy of Daclatasvir/Asun 1097 

Efficacy of Sofosbuvir and S 1206 

Efficacy of combination ther 1157 

Efficacy, Change in MELD Sco 1106 

Evaluation of an Integrated 1136 

Frequency of Renal Impairmen 1099 

Frequent Emergences of Rare 1064 

HCV genotype 3: Meta-analysi 1134 

Health Outcomes and Cost Eff 1089 

Health resource usage in per 1173 

HepCure: An Innovative web-b 1151 

Hepatitis C Cure Could Avoid 1054 

Hepatitis C Cure Results in 1202 

Hepatitis C and HCC Outcomes 1124 

High prevalence of co-morbid 1052 

High ribavirin dose and exte 1201 

Hope and Hopelessness in HCV 1177 

IFN and/or RBV-Free Therapy 1189 

Improving liver function and 95 

Lifetime risks of liver morb 1087 

Lifetime risks of liver morb 1144 

Limited Effectiveness of Sof 1164 

Long-Term Efficacy of Ombita 1086 

Multicenter Experience using 1038 


On-treatment HCV RNA Decline 1130 

Patterns of Care Since the A 1141 

Potential for drug-drug inte 1093 

Predictors of Treatment Adhe 1175 

Preliminary Experience Of Di 1148 

Preliminary Safety and Effic 1065 

Prevalence of drug-drug inte 1131 

Prospective Study for The Ef 1100 

RUBY-I: Ombitasvir/Paritapre 1039 

Rapid drop in serum glucose 1180 

Rapid virological response a 1110 

Real Life Experience of Dire 1104 

Real Life Experience with So 1121 

Real World Effectiveness Of 1188 

Real-Word Effectiveness of L 1053 

Resistance-associated varian 1115 

Resistant-associated variant 1197 

Ribavirin Levels and Transpo 1196 

Safety And Efficacy Of The C 1073 

Safety And Efficacy Of Two I 1147 

Similar Tolerability and Eff 1182 

Sofosbuvir Combination Thera 1184 

Sofosbuvir and Ledipasvir fo 1111 

Sofosbuvir and Ledipasvir in 1037 

Sofosbuvir and Ledipasvir ve 1074 

Sofosbuvir in the Treatment 1090 

Sofosbuvir is well tolerated 1178 

Sustained Virologic Response 1169 

Telaprevir in the Treatment 1112 

The Significance of Immunoal 1059 

The Time and Cost Investment 1172 

The healthcare cost burden o 1068 

The number needed to treat w 1166 

Therapeutic effect of dual o 1159 

Transient renal dysfunction 1113 

Treatment Outcomes of Vetera 1142 

Treatment of recurrent Hepat 1181 

Turquoise-III: 12-Week Ribav 1051 

Twelve Weeks of Sofosbuvir p 1041 

Utility of acoustic radiatio 1195


Utilization and Effectivenes 1135 

Utilizing claims data to und 1085 

Virological efficacy of new 1171 

“Real-life”-Experience w 1183 

Hepatobiliary Neoplasia 

GO1. Experimental 

Hepatocarcinogenesis 

Alpha fetoprotein harbors a 1977 

CD26 (DPP-4) as a molecular 1937 

Cholangiocarcinoma cells int 1993 

Co-activation of AKT and c-M 1934 

Colorectal liver metastasis 1969 

Correlation between Incidenc 1991 

Deregulated Interplay Betwee 1952 

Development of HCC is depend 1971 

EVI1 is a target for gene am 1980 

Efficacy and mechanism of de 1941 

Expression of Transforming G 1962 

FGF15 and FGF19 Induce Dispa 1986 

Fibrotic liver microenvironm 1970 

Glypican 3 (GPC3)-CD81 axis 1979 

Glypican 3 contributes to HC 1967 

HCV Core Gene Mutations with 229 

Hepatic Ruvbl1 is key regula 1963 

Hepatocyte-derived osteopont 1940 

Hyperinsulinemia, not hyperg 1981 

Liver Selective Acetyl-CoA C 1938 

Macrophage colony-stimulatin 1956 

Mevalonate pathway targets F 1974 

MicroRNA profiling of human 1992 

MicroRNA-207 controls Prp19 1964 

Notch-1 Up-regulated Rat Ova 1951 

Oxysterols Promote Malignant 1994 

P300 promotes TGF-β stimula 1947 

Platelet-Derived Growth Fact 1950 

Promoted malignant tumor phe 1968 

Promotion of Hepatocarcinoge 1957 

Steatosis Induces Wnt/Β-Cat 1945 

Survival of hepatocellular c 1960 

The Long Noncoding RNA SNHG6 1939 

The analyses of oxidative DN 1984 

The relationship between hep 1944 

Truncated Hepatitis B virus 1946 

Voluntary exercise opposes a 230 

Baicalein targets liver tumo 1936 

Branched chain amino acids p 1982 

Chronic alcohol accelerates 1958 

Classification of tumors bas 1935 

Continuous hepatocyte apopto 1985 

Critical role of Hippo casca 1948 

Cytotoxic synergy between so 1972 

DDB2 loss protects mice from 1988 

DNA methylome and cancer-spe 1983 

Differential Antitumor Effec 1966 

Hepatocyte-specific overexpr 1949 

Inactivation of Aldose Reduc 1961 

Inflammation contributes to 1989 

Inhibition of autophagy reve 1943 

Lymphotoxin–β regulated b 1987 

Osteopontin is a novel surro 1976 

Osteopontin-c isoform promot 1978 

PPPDE1 is a deubiquitinase t 1973 

PTEN Expression Affects the 1942 

Regorafenib inhibits ERK sig 1953 

Regulator of G-protein signa 1990 

Retrospective cohort study f 1975 

Significance of connective t 231 

Sitagliptin, a Dipeptidyl Pe 233 

Somatic Mutational Analysis 1965 

The Dead Box Protein P68 Can 1954 

Two-step forward genetic scr 234 

Variations of the host genom 1959 

p62 is a Key Regulator for I 232 

GO2. Diagnostics and Liver Imaging 

A mass spectrometry based se 1900 

ARID1A mutations may represe 1901 

Circulating tumor DNA analys 1895 

DNA Methylation Markers for 169 

Diagnostic Performance of Co 1897 

Efficacy of Delta-AFP in Pre 1899 

Evaluation of PIVKA-II effic 1898 

LI-RADS hepatocellular carci 1906 

Transforming acidic coiled-c 1902 

Transjugular Intrahepatic Po 172 

A Model to Estimate Survival 1907 

A Retrospective Cohort Study 1904 

Correlation of MR and pathol 1896 

High 18F-FDG uptake on preop 1893 

Reduced sensitivity of ultra 1894 

The role of AFP in screening 1903 

GO3. Clinical: Hepatocellular 

Carcinoma and Cholangiocarcinoma 

A Single Center Experience w 428 

A Study for the Risk Factors 131 

A potential biomarker, Brf1 460 

A stromal liver gene signatu 364 

Activation of Stemness Genes 366 

Albi score predicts survival 489 

Analyses of progressive dise 482 

Aspirin Use is Associated wi 397 

Cholangiocarcinoma, a signif 416 

Circulating Osteopontin and 170 

Clinical Features and Treatm 486 

Clinical significance of mic 426 

Combining transarterial chem 391 

Comparative Effectiveness of 357 

Comparison of balloon-occlud 411 

Diabetes, HBV infection and 132 

Differences between radiolog 413 

Differences in Presentation 387 

Doxorubicin Resistance in He 352 

Efficacy of Continuous Nucle 439 

Ethnicity impacts aggressive 362 

Etiology-Specific Effect of 415 

Hepatic function and tumour 444 

Hepatocellular Carcinoma Sur 442 

Heterogeneity of advanced he 473 

High gamma-glutamyl transfer 389 

Hong Kong Liver Cancer stagi 409 

Impact of Hepatic p62 Overex 456 

In patients with HCC and mac 421 

In patients with hepatocellu 449 

Influence of hepatitis B vir 405 

Intrahepatic Cholangiocarcin 372 

Lamivudine vs. Entecavir for 422 

Liver microRNA hsa-miR-125a- 431 

Liver-directed (LD) Therapy 481 

MESIAH score is an effective 470 

Meta-Analysis: Proportions o 367 

MicroRNA expression in hepat 429 

Model to predict recurrence 400 

Multi-platform analysis of T 129 

Non-Cirrhotic Hepatocellular 488 

Optimal age to start surveil 350 

Oral nucleos(t)ide analogues 349 

Outcomes following Radiofreq 174 

Outcomes of patients with re 491 

Patients with alcohol compar 373 

Preclinical characterization 394 

Predicting Tumor Death – T 369 

Prediction of Hepatocellular 381 

Prediction of liver decompen 450 

Prognostic Score predicting 467 

Prognostic assessment of pat 355 

Prognostic significance of i 376 

Progranulin Autoantibodies a 463 

Reappraisal of portal vein t 479 

Retreatment with TACE: Trans 447 

Risk Differences of Hepatoce 359 

Risk assessment of hepatocel 346 

Risk factors of complication 171 

Screening Practices for Hepa 401 

Serological markers of extra 441 

Short-term preoperative trea 417 

Significant Suppression of L 410 

Simple serological tests and 475 

Staging Hepatocellular Carci 425 

Stereotactic Body Radiothera 471 

Surveillance at six month in 374 

Survival After Intra-Arteria 344 

Survival analysis of second 472 

TBI 302: A first-in-class th 424 

Targeting the TGF-β Pathway 130 

The Doylestown algorithm – 371 

The autophagy marker LC3 is 398 

The effect of additional tra 464 

The life time frequency and 382 

The role of cytokines and ad 478 

Toronto Hepatocellular Carci 348 

Treatment not needed for hyp 407 

Trends in the burden of cirr 128 

Undetected asymptomatic alco 378 

When to perform surgical res 365 

Whole blood coagulation test 351 

Long term outcome after rese 423 

MicroRNA-26 expression is si 436 

Endothelial nitric oxide syn 435 

A New Clinically-Based Stagi 127 

A Pilot Safety Study of Natu 440 

A randomized controlled stud 402 

Albi score predicts survival 490 

Analytic Morphomics Predicts 418 

Antiviral Therapy for Chroni 207 

Blood alpha-fetoprotein leve 437 

Building of a Scoring Model 419 

Characteristics of etiology- 370 

Clinicopathologic analysis o 465 

Combination of modified Resp 468 

Comparison of Hepatocellular 451 

Comprehensive analyses of mu 406 

Conversion therapy with hepa 399 

Current knowledge of guideli 434 

Development of a model predi 377 

Dysregulated peripheral and 454 

Effect of Antiviral Therapy 368 

Efficacy and safety of mirip 383 

Efficacy of Radiotherapy in 433 

Elevated Serum Wisteria flor 408 

Etiological difference in bo 461 

Exploration of cancer-prone 430


Hepatic Decompensation After 412 

Hepatocellular Carcinoma Can 361 

Hepatocellular carcinoma has 445 

Hepatocellular carcinoma in 462 

Higher Incidence of Hepatoce 458 

Higher Risk for Hepatocellul 388 

Identification of candidate 384 

Identification of novel biom 358 

Impact of Viral Hepatitis Et 483 

Impact of screening on Hepat 404 

Improved Survival of Hepatoc 457 

Integrator complex subunit 6 375 

Korean validation and compar 452 

Liver Transplant Recipients 414 

Long non-coding RNA integrat 353 

Long-term Survival Analysis 474 

Multicentric Study of Liver 469 

Mutations of p53, ARID1A and 347 

Myeloid-derived suppressor c 392 

Natural history of portal ve 393 

Nucleos(t)ide analogue thera 345 

Overexpression of periostin 403 

Plasma MicoRNA-122 level as 379 

Prediction of Recurrence aft 363 

Preventive Effect of Urea-ba 354 

Pro-angiogenic Tie2-expressi 476 

Prospective cohort study for 455 

Racial Differences in the Pr 386 

Racial Disparity in Hepatoce 420 

Regional and Etiological Dif 459 

Skeletal muscle depletion as 477 

Small intrahepatic cholangio 487 

Sorafenib Therapy in HIV-inf 438 

Stereotactic body radiation 484 

Surgical resection versus ra 432 

Surveillance for Hepatocellu 360 

Survival and prognostic fact 485 

Survival of Patients with Ad 395 

Survival of Patients with Ad 448 

The Casein kinase II (CK2) i 396 

The ITA.LI.CA Staging System 356 

The presence of histological 466 

The prognosis and the recurr 453 

The radiofrequency ablation 385 

The relationship between the 443 

Trends of Hepatocellular Car 380 

Validation of the Metroticke 446 

Value of PIVKA II and AFP se 480 

Variability in False-Positiv 427 

Vitamin K dosing during sora 173 

Hepatotoxicity 

HO1. Pathogenesis and Mechanisms 

A comparative analysis of th 595 

A protective role of C/EBP h 592 

Caspase Inhibition Switched 598 

Differentially altered gut m 581 

Golgi Stress Response Is Ass 588 

HMGB1-driven Feedforward Hep 178 

Inhibition of NF-kB by deoxy 176 

Oxaloacetic Acid Protects th 179 

Phenotypic and genotypic cha 583 

Sumoylation Regulates Lipopo 594 

The Clinical-Pathological Sp 597 

The Hepatic “Matrisome” 587 

The influence of drug proper 596 

Transcriptomic Analysis Of H 175 

Acetaldehyde Triggers HCV-In 589 

Critical role of sirtuin 1-m 177 

EPAC mediated protection aga 582 

Effect of Branched Chain Ami 584 

Hormesis in cholestatic live 591 

Mitochondrial Fission Factor 590 

Mitochondrial-shaping protei 599 

Persistent Generation of Inf 585 

Protective effects of connex 586 

STING deficiency protects fr 180 

The Effect of Treatment with 593 

HO2. Drug Metabolism, Toxicity, and 

Therapeutics 

3D culture strategies for im 1921 

AMPK activation prevents and 1909 

Application of the Drug-Indu 1929 

Caffeine Confers Hepatoprote 1917 

Combined Activities of JNK1 1924 

Drug-Induced Liver Injury As 1920 

Gab1 adaptor protein is an e 227 

Gender-specific glucuronidat 1927 

Long-term Outcome of HIV+ Pa 1908 

Protective role of miR-122 a 99 

Severe and prolonged jaundic 1923 

Sodium 4-phenylbutyric acid 226 

Statins prevent liver tumor 1919 

The non-nucleoside reverse t 1913 

Transcriptional Profiling Of 1916 

A Rapid Test for Acetaminoph 1914 

Azathioprine And 6-Mercaptop 1926 

Death and Liver Transplantat 1922 

Drug Induced Liver Injury (D 1928 

Human mercapto-albumin level 1912 

Intervention of RIPK-depende 1918 

Rescue of hepatocytes from l 223 

The role of CD36 in acetamin 1910 

Translocation of Iron from L 1925 

Understanding the Relationsh 1915 

Withaferin-A Reduces Acetami 1911 

HO3. Predictors of Drug Toxicity 

HLA-A*33:01 is strongly asso 225 

Genome-wide investigation id 1933 

Lipotoxicity Accounts for Li 228 

Minocycline hepatotoxicity: 1930 

Predicting In Vivo Hepatotox 224 

Liver Disorders that can Mas 1931 

The Epidemiology of Suspecte 1932 

Human Cholestatic 

and Autoimmune Liver 

Diseases 

IO1. PBC/PSC and Other Cholestatic 

Disease 

Association between elevated 616 

Bezafibrate: A novel and eff 603 

CD39 expression regulates Th 608 

Characterization Of A Patien 630 

Cholestatic liver disease an 631 

Clinical Epidemiology of Pri 75 

Colectomy is associated with 622 

Combination Anti-Retroviral 641 

Elevation of alkaline phosph 634 

Emperipolesis mediated by CD 638 

Enhanced and Prolonged FGF19 604 

Expression of matrix metallo 632 

Gender and IBD phenotype are 76 

Ig Repertoire of Autoantigen 77 

Impact of NGM282 on the Inci 627 

Incidence and Impact of Deco 73 

Increased expression of ORM1 614 

Integrated Analysis of Effic 645 

Long-term follow-up of a mul 605 

Long-term safety and efficac 628 

NGM282, A Novel Variant of F 106 

Patients with Primary Biliar 610 

Patients with childhood-onse 600 

Phenotypic Variations in Pri 607 

S-Alkaline phosphatase is no 633 

Sustained Improvement in the 609 

The IgG/IgG4 mRNA ratio by q 637 

The prevalence and outcomes 626 

Thyroid dysfunction in prima 629 

Validation of serum fibrosis 624 

A Novel Prognostic Model for 74 

Alkaline Phosphatase as Biom 623 

Clinician confidence in stra 636 

Convergence of Two Predictiv 601 

Differentially Expressed Pla 602 

Down-regulation of the secre 621 

Efficacy and Safety of Obeti 625 

Evaluation of the Posology o 635 

Gene expression profiling of 615 

High-risk primary biliary ci 611 

In PSC with dominant bile du 618 

Innate Immunity Drives the I 619 

Intrahepatic Cholestasis of 78 

Long term impact of fibrates 642 

Long-Term Safety of OCA in P 644 

Modeling biliary fluid dynam 617 

Natural killer cells regulat 640 

Phenotypic and Cytotoxic Ana 639 

Predictors of recurrence and 643 

Serum metabolomic profile of 620 

The Modulation of Co-stimula 613 

Unique Immunologlobulin Glyc 606 

Vitamin A deficiency promote 612 

IO2. Autoimmune Liver Disease 

Autoimmune hepatitis type 1 302 

Features and patterns of con 312 

Impact of SNP rs2187668 in H 297 

In autoimmune liver disease 300 

Killer cell immunoglobulin-l 309 

Macrophage migration inhibit 298 

Mean Corpuscular Volume as S 303 

Possible involvement of acti 310 

Soluble PD-1 levels are asso 301 

Tacrolimus or Mycophenylate 308 

Autoimmune Hepatitis Disprop 306 

Carcinogenesis in autoimmune 304 

Immunity in HLA-DR4 expressi 305 

Methotrexate therapy for Aut 311 

Outcomes and mortality of de 307 

Plasma levels of heparanase 299


Inflammation and 

Immunobiology 

JO1. Animal Models 

Novel in vivo and in vitro m 2122 

Allogeneic and xenogeneic tr 2130 

CD8+ T-cells drive liver inj 2128 

Chemokine CCL3/CCR5-recruite 2119 

Chronic ethanol feeding supp 182 

IL-17 signaling in hepatocel 186 

IL-17A plays a pivotal role 2125 

Liver Plasmacytoid Dendritic 2123 

Osteopontin ablation drives 2124 

Small specific-sized Hyaluro 2121 

Small-specific-sized hyaluro 2132 

Sparstolinin B, a TLR4-antag 2133 

Upregulated hepatic expressi 2126 

Vitamin D deficiency promote 2118 

Combined bone marrow plus li 2129 

Hepatocellular carcinoma is 2127 

IFN-α stimulates IFN-γ exp 2120 

The gut-liver axis coordinat 2131 

JO2. Innate Immunity 

Activation of hepatic innate 1290 

Chronic alcohol consumption 1294 

Effects of myeloid cell-sele 1295 

Galectin-3 mediates NLRP3 in 1285 

High baseline expression of 1288 

IL28B genetic variants deter 1287 

Impaired TRAF6 methylation a 1291 

Implants of matrix-embedded 1292 

Kupffer cells signature in a 1286 

Proportion of Intrahepatic C 1289 

The long noncoding RNA VL30- 1284 

Identification of liver mono 1293 

Liver sinusoidal endothelial 184 

Macrophage Specific β-caten 1283 

Mincle Signaling Exacerbates 1282 

The antifibrotic effect of I 183 

JO3. Adaptive Immunity 

Intracellular crawling of ly 1296 

MAIT cells are enriched in p 1298 

Alterations in liver dendrit 1299 

Hepatitis B X-interacting pr 1301 

Inhibition of the FGL2:FcγR 1297 

Soluble CD163 plasma concent 1300 

Toll-like receptor 9 and B-c 1302 

JO4. Mechanisms of Injury 

15-PGDH prevents LPS/GalN-in 185 

Inflammasome Activation Due 2141 

Knockdown of Receptor Intera 2135 

Lack of DNase II exacerbates 2136 

Liver Enzymes Elevation In T 2144 

Macrophage activation throug 2134 

Osteopontin promotes cholang 2143 

Prior Blockade of TNF-α Sig 61 

TRPV4 regulates inflammation 2137 

The attenuation of inflammat 2139 

The gut-liver axis plays a c 181 

Immune Regulation of Mesench 2138 

Role of phagocytosis in coor 62 

TNFα mediates the liver:lun 2142 

The origin and differentiati 2140 

Liver Fibrogenesis and 

Non-Parenchymal Cell 

Biology 

KO1. Basic Fibrosis Research and 

Stellate Cell Biology 

Branched-chain amino acids p 1377 

Cartilage oligomeric matrix 1382 

Characterization of obesity- 1386 

Curcumin blocks the loss of 1412 

Deactivation of human and ra 1418 

Deletion of Wntless in macro 1362 

Deletion of fibrocytes in mi 187 

Dual combination therapy dir 1367 

Efficacy of an ASK1 Inhibito 1359 

Endothelial Niche Derived No 1360 

Essential Roles of RNA-bindi 1396 

Exosome-mediated activation 1358 

HIV infected Kupffer cells m 1397 

Hepatic stellate cell-derive 1414 

Hepatocyte autophagy impairm 1364 

Human precision Cut Liver Sl 1409 

IL-22 enhances TGF-beta pro- 1380 

Identification of Axon guida 1405 

Induction of XBP1 Leads to H 1392 

Inhibition of microRNA-214 a 1395 

Integrin αvβ6 critically r 1368 

Liver fatty acid-binding pro 1361 

MeCP2 exerts global control 1381 

Melatonin Suppresses Activat 1398 

MicroRNA profiling of circul 1404 

Myofibroblastic conversion o 1354 

NLRP3 inflammasome modulatio 1371 

Novel treatment strategy for 1374 

Oral hepatotropic DPP4 inhib 1376 

Paracrine suppression of pro 1416 

Proof of liver regeneration 1375 

Reactive gamma-ketoaldehydes 1407 

Rev-erb and TGF-β Different 1389 

Selective antibody targeting 1379 

Single adenoviral delivery o 1399 

Targeting Activated Stromal 1402 

The AMPK-related kinase NUAK 1410 

The bile acid-phospholipid c 1411 

The paracrine effect of hepa 1373 

Versican: A Novel Modulator 1413 

Interleukin 15 regul 1370 

Antifibrogenic properties of 1391 

Binding of hepatic stellate 1356 

Calcium Mobilization through 1415 

Cancer associated fibroblast 1420 

Circulating exosomes from he 192 

Engineering Healthy And Cirr 1388 

Expression of ENTPD1/CD39 is 1394 

Fibrous network composition 1366 

GP38/PODOPLANIN marks a nove 1385 

Galectin-1 promotes the acti 1400 

Hepatocyte- and hepatic stel 1365 

Human neutrophil peptide-1 e 1421 

IL-10-producing regulatory B 1383 

IL-13Rα1 signaling and M2 m 1353 

Knockout of the substance P 190 

Liver Fibrosis in a Mouse Mo 1422 

Loss of Cytoglobin Exacerbat 191 

Mast cells interact with pro 1363 

Membrane type I-matrix metal 1378 

Mice lacking calmodulin kina 1387 

Modulation of Extracellular 1403 

Molecular mechanism responsi 1390 

Opioid Growth Factor Recepto 1369 

PDGFRα ubiquitination leads 1406 

Potential of connective tiss 1357 

Role of CREBH activation ind 1393 

SNAP-23 Heterozygous Mice Ar 1384 

Selective disruption of dyna 1355 

Sequence Profiling of miRNAs 1408 

Serine Protease Omi/HtrA2 Re 1419 

Splenectomy attenuates advan 188 

TRPC6 Channels Contribute to 1401 

The role of Mesothelin in th 189 

KO2. Imaging and Noninvasive 

Markers of Liver Disease 

Comparison of noninvasive fi 790 

A Novel Diagnostic Index to 774 

Assessment of biliary fibros 788 

Changes in liver stiffness b 777 

Collagen maturation measured 780 

Comparative 10-Years Prognos 786 

ELF Test Thresholds for dise 776 

ELF predicts clinical out 793 

Effect of inflammation on li 791 

Feasibility of three ultraso 802 

Glyco-isomer of Serum Mac-2- 796 

Liver stiffness assessed by 787 

Magnetic resonance elastogra 45 

Non-invasive fluorescent ima 44 

Predicting clinical outcomes 784 

Role of Acoustic Radiation F 800 

Serum Lipidomic Profiling fo 48 

Supersonic Shear Imaging Is 785 

The liver stiffness evaluati 795 

The predictive value of 13C- 46 

Usefulness of Shear Wave Ela 775 

Usefulness of ultrasound ela 781 

qFibrosis scoring for differ 782 

2D Shear Wave Elasto 792 

Differences in Diagn 789 

A Hepatic Stellate Cell Secr 778 

A clustering based fully aut 801 

AnnexinV/EpCAM (CD326) tumor 43 

Association of skin capsular 794 

Coffee consumption reduces l 47 

Diagnostic disparity of FIB4 779 

Fast macromolecular proton f 803 

High risk of misclassifying 773 

Liver fibrosis evaluation us 797 

Machine-learned Image Analys 798 

Real-time shear wave elastog 799 

Serum Wisteria floribunda ag 783 

KO3. Clinical and Translational 

Fibrosis Research 

Activated human hepatic stel 120 

Azathioprine exerts an antif 1445 

Collagen and tissue turnover 1437 

Collagen proportionate area 1440 

Collagen scoring by qFibrosi 1436 

Comparison of 16 blood and/o 1439


Concerted effects of the two 1429 

Correlations between hepatic 1428 

Evaluation of EASL recommend 117 

Host microbiota dictates the 1427 

How gold is the “gold stan 1443 

Innate Lymphoid Cell (ILC) S 1430 

Lysyl oxidase-like-2 (LOXL2) 1442 

PEGylated TRAIL treatment am 115 

Reproductive Aging, Independ 1444 

Risk of cirrhosis in first d 1431 

Serum lysyl oxidase-like-2 ( 1435 

Serum wisteria floribunda ag 1446 

Statins, ACE inhibitors and 116 

Systemic administration of a 118 

The epidemiology of cirrhosi 1424 

Treatment of Hepatic Fibrosi 1423 

A marker of true type III co 1432 

A novel canine liver cirrhos 1447 

A novel glycobiomarker, Wist 1448 

Antifibrotic efficacy of a T 1434 

Comparing the liver stiffnes 1441 

Fibrosis is not just fibrosi 119 

Hepatitis C treatment and Pr 1450 

In vivo cell specific gene s 1426 

In vivo reprogramming of myo 98 

New Prognostic Model for 1-y 1438 

Novel assessment of hepatic 1425 

Safety, Pharmacokinetics, an 1433 

Second harmonic generation m 1449 

Liver Transplantation and 

Liver Surgery 

LO1. Cellular Immunobiology, 

Preservation, and Cell 

Transplantation 

Autologous hepatocyte transp 2 

Blockade of RIP kinase pathw 12 

Delayed Donor Bone Marrow In 54 

Identification of microRNAs 341 

Novel Vasodilator and Anti-i 340 

Previous ischemic preconditi 342 

Extracellular vesicles from 335 

Liver Fabrication Using Dece 336 

Promotion of liver transplan 338 

The innate immune cell micro 339 

Three-dimensional bioactive 337 

LO2. Donor and Allocation Issues, 

Living Donor and Split Liver 

Transplantation, and Hepatobiliary 

Surgery 

3D Printed Liver Models for 840 

A Failure of the Model of En 849 

A Novel Percutaneous Organ P 216 

Application of Gadoxetate Di 869 

Auxiliary partial orthotopic 867 

Clinical impact of 18F-FDG-P 862 

Comparison of Liver Transpla 49 

Controlled attenuation param 876 

Functional Status Predicts P 845 

Impact of UNOS Imaging Crite 10 

Liver Transplant Outcomes an 69 

Liver transplantation waitin 68 

Living donor liver transplan 860 

Living or Deceased Donor Liv 863 

Mechanisms of Gender Dispari 865 

Natural history of liver tra 873 

Older Liver Transplant (LT) 844 

Outcomes of Patients Undergo 854 

Physical function predicts l 841 

Recanalization of complete a 846 

Safety of live liver donatio 859 

The impact of non-identical 72 

Obesity is predictive of 839 

A shift to the left: A compa 838 

Association of Pre-Transplan 855 

Comparable Short And Long Te 70 

Coronary Angiography in Pati 874 

Could Share 35 disadvantage 6 

Effective and Safe postopera 71 

External Validation of a US- 853 

Graft Survival following Dom 868 

Interactive Effect Of Donor 878 

Lack of Radiologic-Pathologi 870 

Long Term Survival Of Liver 875 

Longer Term Efficacy of Simu 836 

Low serum testosterone predi 842 

Novel Sonar Guided Technique 858 

Objective Measurement of Kar 850 

PALBI-An Objective Score Bas 851 

Patient Safety in Living Don 847 

Predicting Futility of Liver 856 

Prediction of waitlist morta 9 

Predictors of Retransplantat 877 

Preliminary Data Analysis fr 837 

Region-Based Projections of 67 

Remnant Liver Ischemia Is As 857 

Significance of functional h 866 

The effect of recipient and 848 

The transportable machine pe 215 

Third Liver Transplantation 864 

Tool for efficient and effec 843 

Traveling for a Liver Transp 872 

Traveling for a Liver Transp 861 

Treatment with Optifast Redu 11 

Using Biomarkers to Predict 871 

Wait Times of 18 Mon 50 

“Oldest Old” (≥80 year 852 

LO3. Immunosuppression, Outcomes, 

and Complications 

15 first-days over-immunosup 1251 

Females, Hispanics, and Nona 1252 

A New Score based on explant 1232 

Aging of liver transplant re 1211 

Are cirrhotic patients await 1239 

Coffee consumption promotes 1222 

Comparative analysis of rena 1233 

Conversion from Sirolimus to 1247 

Downstaging or Bridging Ther 1262 

Early Cytomegalovirus infect 1268 

Early Liver Transplantation 1215 

Everolimus is Associated wit 1226 

Glycine is graft protective 1227 

Good results of liver transp 1269 

How intra-operative factors 1271 

IgG4 status in explanted liv 1246 

Impact of Positive Crossmatc 1254 

Lack of effect of CMV reacti 1230 

Liver Transplant (LT) Candid 1259 

Liver Transplantation For Al 1250 

MELD Based Outcomes of Simul 1249 

Metabolic Syndrome and Insul 1270 

Minimal hepatic encephalopat 51 

Morbid Obesity and Diabetes 1273 

Neurological Syndrome de nov 1256 

Outcomes of Transjugular Int 1280 

Post-Transplant Lymphoprolif 1275 

Pre-Transplant Sarcopenia Do 1267 

Pre-transplant portal vein t 1258 

Prediction of Posthepatectom 1213 

Recipient Risk Factors for A 1266 

Reduced Survival in Elderly 1208 

Reevaluating Risk Factors fo 1241 

Renal Function Improvement i 1281 

Renal function at month 1 an 1237 

Reporting Physical Function 1219 

Retained HLA Class II Donor 1236 

Risk Stratification for Opti 1277 

Role of Coronary Artery Calc 1255 

Rotational Thromboelastometr 1260 

Sarcopenia is a Risk Factor 1276 

Severity and Predictive Fact 1264 

Sublingual administration of 1217 

Synergistic anticancer effec 1278 

Systolic Dysfunction Post-Li 1261 

The Association of Pre-Trans 1242 

The Combination of Strongly 1228 

The Small Molecule TLR9 Anta 1209 

Two-Year Results of a Pilot 1214 

Ursodeoxycholic Acid as Adju 1224 

Outcomes Post-Liver 1243 

A Multi-Center Study to Deve 53 

A Multi-center Study on Down 4 

Acute Kidney Injury among Pa 1220 

Assessment of Energy Metabol 1235 

Blood Phosphatidyl ethanol i 1231 

Echocardiography for Liver T 1257 

Effective Systems of Care Im 1272 

Evaluation of risk indices t 1279 

Incidence of Acute Cellular 5 

Increase in Red Cell Distrib 1240 

Intrahepatic Cholangiocarcin 1274 

Liver Transplantation for Po 1216 

Liver Transplantation in Old 1245 

Long-Term Outcome of Extrahe 1234 

Ohio Solid Organ Transplanta 1265 

Outcomes of Patients with Fa 1263 

Outcomes of acute-on-chronic 1212 

Physical activity measured b 1248 

Pre-transplant echocardiogra 1218 

Primary Biliary Cirrhosis: N 1210 

Quantiferon-CMV testing afte 1225 

Recipient obesity is an inde 1223 

Renal Function Recovery and 1253 

Renal Recovery Outcomes afte 1221 

The Functional Basis for Cog 52 

The Gap Between Assessed Phy 1244 

The coming of age of the gra 1229 

Value of Magnetic Resonance 1238 

LO4. Viral Hepatitis 

Detectable HBVDNA in liver a 1739 

Determinants and impact of e 3 

Effect of simeprevir adminis 1754 

HBV cccDNA evaluation in HBV 1740 

Hepatitis E infection in imm 1752 

Impact of Sofosbuvir-Based R 204 

Liver Cancer in Patients Cur 1742 

Prevention of Recurrent Hepa 200 

Prevention of post-transplan 1751


The effect of ledipasvir/sof 1741 

Two Combined Genetic Markers 1 

An Evidence-Based Algorithm 1745 

Curing Decompensated Wait Li 202 

Current Trends in Liver Tran 199 

Effects of pre-transplant he 1749 

HCV Infection in Aging Baby 201 

Long-term Outcome of Entecav 1746 

Long-term complete prophylax 203 

Long-term outcomes after hep 1748 

Oral Interferon-free Antivir 1753 

Protective Birth Cohort Effe 1750 

Renal safety of nucleos(t)id 1744 

Treating HCV patients on the 1743 

Metabolic and Genetic 

Disease 

MO1. Hemochromatosis, Wilson 

Disease, and a-1 Antitrypsin 

Deficiency 

UGT1A regulation by a member 2108 

Accumulation of hepatitis B 2115 

Association between Psoas Mu 2110 

Clinical features, biochemic 2106 

Congenital glycosylation def 2102 

Dicer-Substrate Small RNAs a 2116 

Liver-specific deletion of t 197 

Mutational analysis of ATP7B 2107 

Pitfalls in Erythrocyte Prot 2111 

Variation in erythrocyte and 2112 

A Nationwide, Population-bas 2113 

Cardiac MRI for Assessment o 2104 

Combining high-throughput ge 193 

Functional analysis and drug 2103 

GNPAT Variant D519G in patie 2117 

Genetic variants associated 2105 

Hepatocellular Carcinoma in 2101 

Liver Fibrosis is Present in 138 

Relapse of Porphyria Cutanea 2100 

Relapse of porphyria cutanea 2114 

The real world experience of 2099 

Wilson disease: gestational 2109 

Midlevel Professionals 

(nurses, nurse 

practitioners, etc.) 

NO1. Behavioral and Quality Issues 

Interferon (IFN), Ribavirin 315 

TNF-alpha is the Most Consis 316 

Longitudinal alteration in h 314 

Monitoring Pruritus for Pati 317 

The Need for Improved Liver 313 

Uncertainty and Hepatitis C: 318 

NO2. Practice Issues 

Significant non–adherence 646 

Hepatology nursing in Canada 647 

Pharmacists mitigate effect 648 

Pediatric Hepatology 

OO1. Pediatric Liver Disease - Basic 

Science 

A novel mouse model for the 2098 

Enteral Obeticholic Acid Pre 194 

Enteral Obeticholic Acid Pro 1689 

Hepatic macrophage IL-1 beta 1687 

Molecular mechanism of IL-1 1688 

OO2. Biliary Atresia and Cholestasis 

B cell activation in the rot 1699 

Biliatresone causes GSH redu 195 

C5aR targets immune and epit 1698 

Exome Sequencing and de novo 1693 

Prophylactic endoscopic trea 134 

Role of transient elastograp 1697 

DXA Bone Density in Alagille 1694 

Diagnostic determination sys 1696 

Expansion of regulatory T ce 1695 

Human iPSC-derived hepatocyt 1690 

King’s-Variceal Prediction 1691 

Molecular genetic dissection 135 

Neurodevelopmental Outcomes 133 

The Rotavirus Induced Mouse 1692 

OO3. Viral and Autoimmune 

Hepatitis 

Autoimmune Liver disease in 1711 

Dissecting cellular entry me 1703 

Prediction of nucleotide ana 1700 

Quantitative Maternal Hepati 1701 

The Impact of Hepatitis B Va 1705 

Validation of transient elas 136 

“Hepatic Granulomas: 18 Ye 1712 

CD4+ helper T Cell Dysregula 1706 

Differentiating autoimmune h 1713 

HCV infection shifts CD8+ T 1708 

Long-term follow up of child 1710 

Pharmacokinetics of Once Dai 1707 

Pre-core region mutations an 1704 

Predictors of poor outcome i 1702 

The Natural History of Autoi 1709 

OO4. Metabolic and Genetic 

Diseases 

NOTCH2 variants in cholestat 1721 

A functional classification 196 

Congenital Lactic acidemia ( 1725 

Liver Involvement in Turnerâ 1720 

Autosomal Recessive Polycyst 1718 

Bile acid metabolism is alte 1726 

Hepatic Manifestations in Ad 1717 

Liver Disease in Subjects wi 1722 

Nocturnal Hypoxia is associa 1715 

Primary immunodeficiencies i 1723 

Recurrent recessive mutation 1719 

Risk Factors Associated with 1716 

Screening for Mitochondrial 1714 

Zinc monotherapy for young p 1724 

OO5. Pediatric Liver Transplantation 

AST-to-platelet ratio index 1733 

Epidemiology and outcomes of 1737 

Preformed and de novo DSA ar 1735 

Serum autoantibodies are ass 1728 

Surveillance biopsies in ped 1736 

Immunization Practic 1738 

Distance Affects Mortality o 1732 

Dynamics of pediatric liver 1734 

Hospitalizations for Respira 137 

Pediatric liver transplant r 1730 

Pre-transplant mortality ris 1727 

Racial and ethnic variations 1729 

Regulatory T cells in de nov 1731 

Portal Hypertension and 

Other Complications of 

Cirrhosis 

PO1. Experimental 

Adipose Tissue-Specific ATGL 1531 

Diabetes is Associated with 1527 

ELF and Angiopoietins acc 1530 

Elevated galectin-3 in cirrh 1529 

Enoxaparin treatment does no 1520 

Hepatocyte Tissue Factor Con 81 

Inflammation and hypoxia reg 1519 

L-carnitine prevents ammonia 1528 

Liver NK cells from NLG4-/- 1532 

Long term oral treatment of 1523 

Neuro-inflammation in Cirrho 1521 

Portal tract fibrosis is ind 1526 

Simvastatin prevents the agg 1518 

Statins activate the canonic 80 

Taurine lowers portal pressu 149 

The Evaluation of Portal Hyp 1524 

The effects of apoptosis inh 79 

Ammonia Produces Pathologica 1517 

Emricasan, a pan caspase inh 1522 

The soluble guanylyl cyclase 1525 

Usefulness of spleen stiffne 1533 

PO2. Ascites, Renal Dysfunction, and 

Hepatorenal Syndrome 

Acute Kidney Injury definiti 266 

Acute Kidney Injury in criti 275 

Acute kidney injury in cirrh 288 

Chronic Rifaximin Therapy De 271 

Clinical Utilization of Seru 280 

Coronary microvascular dysfu 268 

Deleterious effect of beta-b 265 

Diagnostic accuracy of the P 264 

Disturbance in diurnal renal 267 

Effect of Beta-Blockers on S 282 

Is Midodrine, Octreotide, an 286 

Midodrine and Tolvaptan in P 259 

Multicenter retrospective an 281 

New score CyBAC-Del incorpor 262 

Organ Failures Associated Wi 83 

Predictive factors for a goo 296


Predictors of response and s 278 

Serelaxin increased renal bl 260 

Slow, continuous low-dose al 274 

Terlipressin given by contin 148 

The Trigger Matters – Outc 270 

The association of non-selec 82 

Urine cystatin C is a strong 277 

Acute kidney injury and shor 289 

Ascites Neutrophil Gelatinas 272 

Ascites neutrophil dysfuncti 261 

CA-125 significance in cirrh 295 

Efficacy and safety of intra 279 

Goal-Directed Treatment for 290 

Hepatiq Measure Of Hepatic F 269 

Increasing incidence and cos 283 

Is prevalence of spontaneous 285 

Low ascites levels of cell m 276 

Renal Biopsy Histopathologic 284 

Signicant Ascites in Childre 294 

TIPS with PTFE-covered stent 263 

The decreased urinary aquapo 287 

Use of non-selective beta bl 273 

Usefulness of the hepatic ve 291 

Validation of Ascites-specif 293 

What is the evolution of Gra 292 

PO3. Varices and Bleeding 

An early experience of the B 764 

A Randomized, Multi-center, 736 

Atorvastatin and propranolol 750 

Balloon-occluded retrograde 748 

Clinical and histologic corr 737 

Correlation between noninvas 739 

Direct oral anticoagulants h 761 

Does Haemodynamic or Clinica 772 

External validation of risk- 742 

Hemostatic profiles in liver 740 

Inter-observer variability i 744 

Ischemic hepatitis following 146 

Obesity is associated with i 735 

Occlusion of portosystemic s 760 

Prevention of Recurrent Vari 745 

Propranolol for Prevention o 733 

Risk factors for in-hospital 765 

Serelaxin reduced portal pre 753 

Serum lysyl oxidase-like-2 ( 746 

The impact of esophagogastri 759 

The predictive value of chan 771 

Transient Elastography and s 755 

Transjugular intrahepatic po 769 

Viral suppression with IFN-f 732 

Transient elastograp 762 

A prospective study comparin 752 

Acute kidney injury predicts 757 

Argon Plasma Coagulation Ver 758 

Baveno`s VI non-invasive app 84 

Comparison of Beta blockers 766 

Effect of nonselective beta- 768 

Efficacy and safety of treat 767 

Efficacy of carvedilol as an 738 

Establish risk scores for pr 751 

Factors Influencing The Firs 734 

Long-term results of balloon 756 

Placebo-Controlled, Randomiz 743 

Prospective validation of th 749 

Shunt or meso-portal bypass 763 

Spleen stiffness assessed wi 741 

The relationship between the 770 

Transjugular intrahepatic po 754 

PO4. Encephalopathy and Other 

Complications 

Acute decompensation of cirr 1482 

Ammonemia predicts severity 1510 

Anticoagulation in patients 150 

Autonomic cardiac dysfunctio 1495 

Bioelectrical impedance vect 1471 

Bromocriptine is effective i 147 

Characteristics, indications 1508 

Clinical Characteristics of 1464 

Conjugated bile acids suppre 1507 

Efficacy of Ornithine Phenyl 1509 

Erectile Dysfunction in pati 1476 

Gut Microbiota Alterations a 1463 

Increasing frequency of gram 1489 

Liver Injury Independently P 1488 

Low Serum Leptin Level Is A 1496 

Minimal hepatic encephalopat 1502 

Muscle mass in liver transpl 1468 

Myocardial strain as a nonin 1494 

Obesity Paradox: Obesity is 1469 

Perceptions of the ‘cirrho 1477 

Plasma Metabolomic Profiling 1470 

Portal vein thrombosis, mort 1501 

Preoperative plasma glucose 1492 

Progressive Loss of Hematopo 1483 

Proliferative lymphangiogeni 1514 

Real Time Pressure Volume Lo 1503 

Role of 100 % FiO2 PaO2 and 1513 

Sarcopenia and Sarcopenic-Ob 1481 

Six minute walking test perf 1484 

Six-week administration of l 1475 

Transient elastography relat 1466 

Transjugular Liver Biopsy Is 1498 

Use of betablockers, previou 1511 

Usefulness of Balloon-Occlud 1486 

Usefulness of Balloon-Occlud 1474 

Validation of Relationship b 1504 

Videogame Training Improves 1491 

Sarcopenic obesity i 1500 

A half of hepatic hydrothora 1499 

Comparison Of Thrombin Gener 1505 

Development of a Novel Clini 1485 

Development of an Electronic 1467 

In Vivo Assessment Of Portal 1506 

Inadequate Food And Water In 1493 

Is it safe for patients with 1472 

Lactulose treatment improves 1465 

Linkage and interaction betw 1516 

Myocardial blood flow is red 1480 

Non-invasive assessment of p 1490 

Osteopontin is a new prognos 1487 

Prospective Multicenter Obse 1478 

Randomized Controlled Trial 1473 

Relative Adrenal Insufficien 1515 

Sarcopenia predicts moratili 1479 

The World Congress of Gastro 1497 

The burden of osteoporotic h 1512 

PO5. Infections and Acute on Chronic 

Liver Failure 

25-hydroxyvitamin D deficien 2086 

Adding C-reactive Protein an 2089 

Cholangitis is a Leading Com 2084 

Circulating interleukin 6, 1 2075 

Comparison And Outcomes Of 5 103 

Effects of norfloxacin thera 145 

Lack of evidence for bacteri 2094 

Liver cirrhosis and short-te 2095 

Neutrophil to Lymphocyte Rat 2085 

Obesity is a Major Predictor 2081 

Polymorphisms in the interle 2087 

Presepsin as a new biomarker 2090 

Prevalence, risk factors and 2070 

Sepsis in Cirrhosis Patients 2093 

Short-term infusion of soyab 2088 

The Epidemiology and Clinica 2073 

The NDP52 Val248Ala variant 2077 

The macrophage activation ma 2071 

The relationship of fecal ba 2091 

A Broad Spectrum Ant 2067 

A prognostic model evaluatin 2096 

Assessment and relevance of 2078 

Characterisation of blood mi 2074 

Characteristics and outcome 2079 

Hispanic ethnicity is associ 2083 

Impact of genetic variation 2069 

Liver Cirrhosis Is Strongly 2082 

MELD score and liver stiffne 2080 

Pneumonia Outcome Score In C 2092 

Probiotic supplementation im 2068 

Sequential assessment of Acu 2072 

Systemic inflammatory respon 2076 

The relationship between pro 2097 

Steatosis and 

Steatohepatitis 

QO1. Steatohepatitis: Experimental 

A DPP-4 inhibitor, sitaglipt 966 

NorUDCA reduces liver injury 977 

The obese insulin resistant 978 

A Balanced Carbohydrate and 950 

Association between nonalcoh 964 

Adipose tissue-derived strom 947 

Alteration of Neutrophil-der 898 

An Intact Fgf15 Signaling Pa 916 

Ancestral starvation-activat 936 

Angiogenesis in the pathogen 954 

Anti-Fibrotic Effect of Auto 141 

Assessment of Donor and Envi 915 

Beneficial effects of obetic 893 

Blocking of IL-17A signaling 890 

CHOP and the Unfolded Protei 884 

Cholesterol crystallization 923 

DSS colitis has tumorinogene 967 

Deficiency of intestinal muc 59 

Deletion of G-protein-couple 885 

Differential carbonylation o 921 

Dual targeting of nuclear re 897 

Ectopic expression of N-acet 906 

Ethanol-Mediated Lipocalin 2 879 

Fatty Acid-Stimulated Inflam 935 

Fenofibrate and LXRα agonis 924 

Fermented soymilk prevent fr 951 

FoxO3 increases miR-34a to c 60 

Free fatty acid treatment re 975 

GFT505 (ELAFIBRANOR) prevent 974 

Glucagon-like peptide-1 pote 955 

Hepatic Estrogen Receptor Ne 956 

Hypoxia accelerates fatty ac 930 

Inhibition of Mitophagy in N 968 

L-carnitine ameliorates hepa 917 

Lipid-induced Endoplasmic Re 883


Liver-Directed Allosteric In 957 

Metformin targets a phosphoS 958 

MicroRNA-21 is a Potential L 102 

Mixed Lineage Kinase 3 Media 895 

NADPH oxidase (NADPHox) play 896 

NFkB inhibition by Andrograp 969 

NOX2 deficiency attenuates e 910 

Non-invasive quantitative de 913 

Nuclear localization of auto 891 

Omentectomy prevents nonalco 939 

Over-expression of HNF4α at 941 

Peretinoin, an acyclic retin 144 

RIP3-dependent hepatocyte ne 940 

Reduction of Hepatic 27-Hydr 970 

Regulation of hepatocellular 929 

Regulatory T Cells Prevent A 909 

Role of Fatty Acid Desaturas 56 

Role of Fcgamma receptors in 971 

Role of the receptor tyrosin 973 

S-Adenosylmethionine Treatme 953 

Soluble Intercellular Adhesi 892 

Standard American Diet (SAD) 946 

Targeting enterohepatic bile 140 

The dual FXR/TGR5 agonist IN 882 

The dual FXR/TGR5 agonist IN 902 

Vitamin D through Induction 904 

miRNA-339-3p is increased in 960 

L-Fabp deletion attenuates b 911 

A novel transcriptional repr 925 

Aging associated impaired me 901 

Akkermansia Muciniphila Is D 932 

Aliskiren reduces liver and 965 

Alterations in NK cell pheno 938 

Analysis of T and myeloid ce 928 

Antihypertensive therapy imp 937 

Bile Acid Sequestrant Preven 139 

Blocking the NLRP3 inflammas 55 

CX3CR1 is a gatekeeper for i 100 

CXCL1 promotes steatohepatit 903 

DNA Methylation Profiles of 889 

DRX-065, the stabilized R-en 143 

Deletion of orphan nuclear r 920 

Effects of Dietary Different 926 

Enhanced expression of Rubic 97 

Exacerbation of non-alcoholi 959 

Fecal Alkaline Phosphatase I 905 

Genetic Ablation of Phosphat 900 

Genetic analysis of NAFLD wi 943 

Hepatic NOD-like receptors, 886 

Hepatic but not Intestinal S 919 

Hepatocyte-specific autophag 894 

Hepatocyte-specific depletio 880 

Hepatocytes Release Ceramide 58 

High Milkfat Diet Causes Mor 907 

High and Low Capacity Runner 948 

High calories intake and par 949 

Human amniotic epithelial ce 945 

Increased 53-binding protein 952 

Increased sensitivity and co 976 

Inhibition of β-hydroxybuty 979 

Inhibitor Of DNA Binding 2 G 944 

Key role of ASMase in the su 899 

Leucine synergizes with metf 933 

Liver MicroRNA-21 is Overexp 887 

Liver sinusoid endothelial c 927 

Lysosomal cholesterol in Kup 922 

Mast Cell Involvement in Non 914 

MiR-128-3p is enriched in th 57 

Ornithine transcarbamylase g 908 

Predicted prevalence of NAFL 931 

Regulation of Adaptive Therm 881 

Regulation of Hepatocellular 972 

Role of fibroblast growth fa 934 

TRPV4 deficiency enhances TL 962 

Thioesterase Superfamily Mem 942 

Treatment with the FXR-TGR5 142 

Two different aspects of PD- 888 

Western diet for 7 weeks in 961 

Wheat Bran Autolytic Peptide 918 

Intrahepatic expression leve 912 

β7-Integrins and MAdCAM-1 h 963 

QO2. Steatohepatitis: Clinical and 

Therapeutic 

A Meta-analytic Assessment o 2212 

A novel noninvasive diagnost 2156 

ANA Risk (Age-NAFLD-Apri Ris 2248 

Adults With Nonalcoholic Ste 237 

Application of transient ela 238 

Are the phenotypes of nonalc 2223 

Assessment of parameters rel 2254 

Association of Nonalcoholic 2219 

Beneficial effects of the du 162 

Can FibroMeter VCTE be us 2180 

Can we avoid liver transplan 2215 

Cenicriviroc and Pioglitazon 2247 

Characterization of insulin 2239 

Clinical features and risk f 2171 

Combination of serum HA, CK1 2160 

Combined effects of the pros 2195 

Controlled Attenuation param 2181 

Differences of lifestyle beh 2243 

Digoxin provides hepatoprote 2179 

Discovery of plasma biomarke 2232 

Effect of biopsy length on h 2153 

Efficacy and Safety of Vitam 107 

Exercise reduces liver fat, 2183 

Fibrometer Nonalcoholic Fatt 2235 

First degree relatives of su 2203 

Hepatic expression of the ap 2149 

High prevalence of undiagnos 2231 

High prevalence of PNPLA3 rs 2210 

Hypovitaminosis D in NAFLD r 2186 

IL-8 and MCP1 are Independen 2169 

Identification of Unique Mar 2194 

In Female Patients with Non- 2205 

Increasing severity of NAFLD 2188 

Liver Fat and Inflammation A 2187 

Longitudinal changes in FIB- 239 

MHC Class I Related Gene A ( 2173 

Macrophages (CD68+ Cells) an 2192 

Mechanism of Action of the A 2246 

Mitochondrial Mutator-Phenot 2217 

Molecular Characterization o 2221 

Naltrexone/Bupropion Extende 2175 

Non-Alcoholic Fatty Liver Di 2225 

Non-Invasive Estimation of D 2206 

Non-alcoholic Fatty Liver Di 2204 

Non-alcoholic fatty liver di 2238 

Non-invasive Diagnosis of No 2151 

Non-invasive hepatic fibrosi 2191 

Normal body mass index as a 2251 

Obesity fibrosis score: A ne 2182 

Older Age, Increasing Body M 2159 

Omega-3 fatty acids improve 240 

Outcome of bariatric surgery 2222 

Plasma Metabolomic Profiles 161 

Potential benefit of folate 2162 

Predictors of improvement in 2154 

Prevalence and severity of n 235 

Prevalence of hepatic steato 2253 

Prevalence of non-alcoholic 2189 

Relationships between very l 2228 

Relevance of the Oxidative S 2237 

Reverse correlation with FXR 2218 

Risk factors for hepatocellu 2166 

Serum marker of inflammasome 2185 

Short Sleep Duration is asso 2155 

Steatohepatitis and liver fi 2227 

Substantial variability of t 2164 

Subtyping nonalcoholic steat 2244 

Testosterone Is Associated w 2252 

The dual and opposite role o 2216 

The effectiveness of SGLT-2 2240 

The impact of alcohol consum 2241 

Transient Elastography is Fe 2220 

PNPLA3 and JAZF1 variants in 2158 

1H-Magnetic Resonance Spectr 2152 

A 32-gene poor prognosis sig 2174 

An international, phase 2 ra 105 

Changes in plasma biomarkers 2161 

Characteristics and Clinical 2250 

Clinical and Metabolic Effec 236 

Concomitant Non-Alcoholic Fa 2214 

Cross-sectional and longitud 2150 

Cross-sectional and longitud 2201 

DACRAs are novel therapeutic 2178 

DJ-1 labeling index is a nov 2165 

Diacylglycerol acyltransfera 2208 

Diagnostic accuracy for non- 2177 

Disturbance of regulatory T 2167 

Effect of Pradigastat, a Dia 2147 

Extending the Ballooning Sco 157 

Functional inhibition of spl 2198 

Haptoglobin Genotype 2 Allel 160 

How common is chronic liver 2168 

Importance of reduction in s 2202 

Improvement in Hepatic Steat 2200 

Increased Immune Responses o 2157 

Longterm Effects of Bariatri 2170 

Minimally deranged serum ala 2226 

Multi-parametric magnetic re 2190 

Non-alcoholic fatty liver is 2245 

Overweight in late adolescen 158 

Plasma Cytokeratin-18 Fragme 2148 

Polychlorinated biphenyl exp 2233 

Prebiotic supplementation wi 2176 

Preserved hemostatic status 2249 

Prospective comparison of no 2211 

Risk factors for hepatocellu 2207 

Risk factors for progression 2213 

Serum oxidative-anti-oxidati 2236 

Subjects with non alcoholic 2209 

The Association Between Sarc 2234 

The Lipidomic Signature Of D 2163 

The Performance of Vibration 2197 

The association of ALT level 2224 

The development of hepatocel 2230 

The glycosylation marker M2B 2199 

The hepatic and extra-hepati 2145 

The link between intestinal 2184 

Upregulated absorption of di 2146 

Usefulness of mac-2 binding 2193 

Weight Loss Results in Signi 2196 

Zone 1 and Zone 3 Steatosis 159


QO3. Alcohol: Clinical and 

Experimental 

Alcohol modifies the composi 1311 

An alarmin cytokine IL-33 in 1345 

Antimicrobial proteins Reg3b 111 

Ceramide Synthase May Regula 1314 

Cross talk between hepatocyt 1339 

Differential Changes In Pro- 1309 

Differential microRNA expres 1332 

Elevated leukocyte count com 1350 

Ethanol Exposure Inhibits He 1310 

Ethanol-impairs mTOR activit 1308 

Fat-specific Protein 27/CIDE 110 

Functional role of the Lin28 1349 

Genetic Ablation of MitoNEET 1338 

Hepatocyte-specific StARD1 d 1320 

Hepatocytes from mice on int 112 

Impaired Proteasome Function 1325 

Low Dose Zinc Sulfate (220mg 1303 

Microparticles Are Markers O 1344 

Molecular chaperone Hsp90 re 1351 

Nogo-B facilitates M1 polari 1322 

Positive familial metabolic 1328 

Role of Pre-mRNA Splicing Re 1324 

The Alcoholic Hepatitis Hist 1337 

The Epidemiology and Costs o 1329 

Short Term Abstinenc 113 

Creatine Supplementa 1347 

Ethanol-Induced Alte 1323 

Acrolein, a lipid-derived al 1304 

Alcohol stimulates macrophag 1315 

Alterations of gut microbiom 114 

Characteristic features of m 1326 

Coffee Consumption Protects 1316 

Cyr61 protection of hepatocy 1352 

Early prediction of response 1307 

Effects of Alcohol and Absti 1343 

Fibroblast growth factor 21 1306 

Gut Bacterial Load Influence 1330 

Hepatocyte-specific deletion 1313 

Increased IL-17 production i 1318 

Infection in alcoholic hepat 1321 

Inflammatory and apoptotic m 1334 

Interplay between microRNA-2 1317 

Intestinal HIF-1α Deletion 1305 

LPS-TLR4 pathway mediates du 1327 

Macrophage Migration Inhibit 1346 

Microbiota protects mice aga 1319 

Nosocomial Infection in Seve 1342 

Nuclear receptor Rev-Erbα f 1336 

Oral pentamidine (VLX103) pr 1333 

PDE4 inhibition attenuates a 1312 

Sarcopenia and adipopenia se 1340 

The L-carnitine alleviate he 1348 

The role of proinflammatory 1341 

Tributyrin, a butyrate pro-d 1331 

Zinc deficency inactivate he 1335

studies

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