2008 Barcelona - European Society of Human Genetics

Therapy for genetic disease
in cells bearing the following genotypes: L444P;E326K/G202R (1 .3fold
at 2.5 μM), D409H/N188S;E326K (1.4-fold at 5μM and 10μM, and
1.2-fold at 20μM), and N370S/N370S (1.7-fold at 5μM and 10 μM, and
1.5-fold at 20μM). Treatment with the aminocyclitol C10 at different
concentrations increased 1 .5-fold the GBA activity in L444P/G202R
fibroblasts, and from 1.2 to 1.7-fold in L444P;E326K/G202R patient
cells .
P10.13
Hematopoietic stem cells therapy and risk of graft versus host
disease: A report from iran
M. Ebrahimi 1 , M. Houshmand 2,1 ;
1 Special Medical Center, Tehran, Islamic Republic of Iran, 2 National Institute of
Genetic Engineering & Biotechnology, Tehran, Islamic Republic of Iran.
Transplanted hematopoietic stem cells (HSC) and progenitor’s cells
can treat malignant and nonmalignant disorders, including immunological,
gynecological, neurological, endocrinological and others
pathologies and disorders . The transplantation of HSC that was not
genetically identical (allogeneic)to that of the recipient resulted in an
immunologic reaction by the donor lymphocytes against the recipient,
causing inflammation of the target tissues, termed graft-versus-host
disease (GVHD) . GVHD is one of the major limiting factors in successful
HSC transplantation . A wide range of host antigens can initiate
graft-versus-host-disease, among them the human leukocyte antigens
(HLAs) . HLA-identical siblings or HLA-identical unrelated donors often
have genetically different proteins that can be presented by MHC molecules
to the recipient’s T-cells, which see these antigens as foreign
and so mount an immune response .
we enrolled 18 patients with one form of neuromuscular disorders at
Genetics Department of Medical Special Center in Tehran .The all patients
received transplants of HSC of the human embryonic liver . All
patients were treated on clinical protocols, which were reviewed and
approved by the Embryotech and Special Medical Centers . All patients
provided writing informed consent before being enrolled in the protocols
.
The diagnosis of acute GVHD was initially based on clinical signs must
be confirm by positive biopsy results from at least one involved organ.
In period of observation GVHD is developed in nobody . The lowest risk
of GVHD is associated with that the HSC derived from liver do not contain
the antigens of major histocompatibility complex on their surface
what makes them tolerant towards recipients .
P10.14
Effects of silibinin on cell growth and invasive properties
of a human hepatocellular carcinoma cell line, HepG-2,
through inhibition of extracellular signal-regulated kinase 1/2
phosphorylation
M. Noori-Daloii, M. Momeny, M. Khorramizadeh, M. Yousefi, M. Yekaninejad,
R. Esmaeili, Z. Jahanshiri, A. Noori-Daloii, S. H. Ghaffari;
Tehran Univ. of Medical Sciences, Tehran, Islamic Republic of Iran.
The purpose of the current study is to evaluate the effect of silibinin on
human hepatocellular carcinoma HepG-2 cells . MTT assay, LDH release,
Gelatin zymography, Griess reaction, Cell-based ERK 1/2 phosphorylation
assay and quantitative real-time RT-PCR were employed
to appraise the effect of silibinin on cell proliferation, cytotoxicity, metastatic
potential, nitric oxide (NO) production, ERK 1/2 phosphorylation
and activation in HepG-2 cells . Silibinin inhibited cell proliferation, matrix
metalloproteinase 2 enzymatic activity, NO production and ERK
1/2 phosphorylation in a dose-dependent manner without exerting any
cytotoxicity effect . In addition, an expressive increase in mRNA levels
of Raf kinase inhibitor protein (RKIP), sprouty-related protein 1 with
EVH-1 domain (Spred-1), sprouty-related protein with EVH-1 domain
2 (Spred-2) and tissue inhibitor of matrix metalloproteinase 2 (TIMP-2)
coupled with a significant reduction in transcriptional levels of highly
expressed in cancer (Hec1) and MMP-2 were observed . Altogether,
these issues show for the first time that silibinin treatment could inhibit
cell proliferation and invasive potential of HepG-2 cells through inhibition
of ERK 1/2 cascade both directly (through suppression of ERK 1/2
phosphorylation) and indirectly (through up-regulation of RKIP, Spred-
1 and Spred-2) . In addition, cell growth and proliferation may be inhibited
by silibinin through down-regulation of Hec1 .
P10.15
Evaluation of transient transfection methods in Hu11 hybrid
cells
A. Asgharian 1 , J. Gharesouran 2 , Z. Deilami Khiabani 1 , H. Najmabadi 2 , M.
Banan 2 ;
1 Cell and molecular biology,Islamic Azad University, Tehran, Islamic Republic of
Iran, 2 Genetics Research Center, University of Social Welfare & Rehabilitation
Sciences, Tehran, Islamic Republic of Iran.
One important step in expression of exogenous genes in cells is transfection
. The Hu11 cell line is a mouse erythroleukemia (MEL) cell line
containing the human chromosome 11 (and thus the β-locus). Hu11
cells express the human globin genes and thus should useful for studying
the basis of globin gene regulation . In this study, we tested several
transient transfection methods in Hu11 hybrid cells. The PSV-β-Galactosidase
vector was used as a control to monitor transfection efficiency.
We tested commercially available reagents such as lipofectamin
TM 2000 (Invitrogen), FuGENE ® HD transfection Reagent (Roche),
X-tremeGENE siRNA transfection Reagent (Roche), HiPerfect Transfection
Reagent (Qiagen) and DEAE-Dextran (Sigma) . Also Hu11 cells
were electroporated under different conditions . We have shown that
different cationic lipid transfection reagents do not provide a reliable
and effective method of transfecting Hu11 cells . Also electroporation
did not work well for this cell line . Therefore we suggest that the best
method for efficient transfection of Hu11 cells can be accomplished via
a viral vector-based transfection procedure .
P10.16
characteristics of patients with Hunter syndrome in spain and
Portugal compared with those in the rest of the world: analysis
of data from HOs - the Hunter Outcome survey
G. Pintos-Morell 1 , E. Leão Teles 2 , M. del Toro Riera 3 , M. Beck 4 , R. Giugliani 5 ,
R. Martin 6 , J. Muenzer 7 , E. Wraith 8 ;
1 University Hospital ‘Germans Trias i Pujol’, Badalona, Spain, 2 Sao Joao Hospital,
Porto, Portugal, 3 Hospital Vall d’Hebron, Barcelona, Spain, 4 University
of Mainz, Mainz, Germany, 5 Medical Genetics Service, HCPA/UFRGS, Porto
Alegre, Brazil, 6 Saint Louis University, St Louis, MO, United States, 7 University
of North Carolina, Chapel Hill, NC, United States, 8 Royal Manchester Children’s
Hospital, Manchester, United Kingdom.
Aims: To compare the characteristics of patients with Hunter syndrome
(mucopolysaccharidosis type II) in Spain and Portugal with patients in
the rest of the world (ROW) .
Methods: Analysis of data from HOS - the Hunter Outcome Survey
- was conducted in January 2008 . HOS is a global outcome survey
established to assess the natural history of Hunter syndrome and the
safety and effectiveness of enzyme replacement therapy with idursulfase
(Elaprase ® ; Shire HGT, Danderyd, Sweden) . As of January 2008,
there were 367 ‘prospective’ patients included in HOS, 33 of whom
were from Spain and Portugal .
Results: Mean age (±SD) at onset of symptoms in patients from Spain/
Portugal was 1 .7±1 .1 years and from the ROW 2 .1±1 .8 years . Delay
in diagnosis after symptom onset in Spain/Portugal was markedly less
than that for the ROW (1 .1±1 .4 vs 2 .2±2 .9, respectively) . The occurrence
of any neurological signs/symptoms was similar (84%) in patients
from Spain/Portugal and the ROW . Respiratory symptoms were
reported in 75% of patients in Spain/Portugal and in 84% of patients
from the ROW . Cardiovascular signs/symptoms were reported in 69%
and 85% of patients from Spain/Portugal and the ROW, respectively .
Characteristic facial features were the most commonly reported manifestation
of Hunter syndrome, occurring in over 90% of patients in
Spain/Portugal and the ROW .
Conclusions: This analysis of HOS data indicates no substantial difference
between patients with Hunter syndrome in Spain/Portugal and
the ROW . However, it highlights the delay between the occurrence of
signs/symptoms and diagnosis, and the need for increased awareness
of this rare disease .
P10.17
Optimization of transient transfection of K562 in order to siRNA
transfection
A. Asgharian 1 , J. Gharesouran 2 , Z. Deilami Khiabani 1 , H. Najmabadi 2 , M.
Banan 2 ;
1 Cell and molecular biology,Islamic Azad University, Tehran, Islamic Republic of
Iran, 2 Genetics Research Center,University of Social Welfare & Rehabilitation