Haematologica 2003 - Supplements

More documents

Recommendations

Info

8. Mouse models for MM 214 Suppressor of Cytokine Signalling-3 (SOCS3) is a key physiological negative regulator of IL-6 Andrew Roberts, Ben Croker, Danielle Krebs, Jian-Guo Zhang, Sam Wormald, Don Metcalf, Nicos Nicola, Douglas Hilton, Warren Alexander The Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia. Because IL-6 is a key modulator of myeloma cell survival and proliferation, identification of intracellular negative regulators of IL-6 signalling is an important step in the process of developing novel therapies. The SOCS family of proteins are attractive candidates as physiological regulators of IL-6 signalling. SOCS1 and SOCS3 are highly homologous and the expression of both is induced by IL-6 in both lymphoid and myeloid cells. In overexpression systems, both can inhibit STAT activation after gp130 ligation by IL-6. However, IL-6 signalling is not perturbed in primary cells from SOCS1 knockout mice. We have therefore concentrated on investigating the role of SOCS3 in IL-6 signalling in primary haemopoietic cells. A conditional gene targeting strategy was pursued because SOCS3 knockout mice die early in utero of placental failure. We have generated mice in which either haemopoietic or hepatic cells have no functional SOCS3 alleles, by using mice bearing a Lox-P flanked allele of SOCS3 (ki) and a null allele (o), and intercrossing them with mice expressing Cre recombinase selectively in haemopoietic cells (LysMCre or VavCre) or hepatic cells (AlbCre) respectively. Hepatocytes from AlbCre+SOCS3fl/o do not express SOCS3. Similarly, macrophages from LysMCre+SOCS3fl/o mice and all haemopoietic cells from VavCre+SOCS3fl/o mice are totally deficient in SOCS3. In all SOCS3 deficient cells from these mice, marked abnormalities in IL-6 signalling and cellular responses were observed. Following either in vitro or in vivo stimulation with IL-6, STAT3 phosphorylation was both increased and prolonged. Microarray analysis confirmed this excess signalling resulted in aberrant target gene transcription. Accordingly, cellular responses to IL-6 were amplified in the absence of SOCS3: (i) serum acute phase proteins were increased in AlbCre+SOCS3fl/o mice; (ii) IL-6 inhibition of macrophage proliferation was augmented; and (iii) myeloid progenitor cell proliferation was increased. These data unequivocally prove that SOCS3 is a key physiological negative regulator of IL-6 signalling. Whether the absence of SOCS3 increases plasmacytoma development or progression in murine models is being addressed. 215 NOVEL TARGETED DEREGULATION OF APOPTOTIC AND ONCOGENIC PATHWAYS LEADS TO ACCELERATED B-LYMPHOID MALIGNANCY IN GENETICALLY ENGINEERED MICE Michael Linden* Wan-Cheung Cheung#, Sung Sup Park$, Nicole Kirchhof*, Cathy Carlson* Kathy Pape#, Marc Jenkins#, Roberto Polakiewicz#, Siegfried Janz$, and Brian Van Ness* #Cell Signaling Technologies (Beverly, MA), $National Cancer Institute (Bethesda, MD), *University of Minnesota-Twin Cities (Minneapolis, MN) Multiple myeloma (MM) is characterized by the proliferation of malignant plasma cells (PC’s) in the bone marrow. We have undertaken a collaborative project to target gene deregulation that will contribute to models of plasma cell malignancy in the mouse. Specifically, our project seeks to address three goals: 1) Identify novel transcriptional regulators whose activity is limited to B- lymphoid cells in late developmental stages; 2) Develop new mouse models of B-lymphoid malignancies, including MM, using transcriptionally targeted transgenes; and 3) Develop novel strategies to traffic plasmacytomagenesis in vivo. While no pathognomonic lesion has been identified in MM, upregulation of anti-apoptotic proteins (BCL-XL) and deregulation of growthpromoting oncogenes (MYC and N-RAS) are common in many MM patients. The 3' kappa immunoglobulin (Ig) light chain enhancer (3’KE) regulates transcription of the kappa Ig gene, and its activity in murine B-cells is restricted to the late stages of B- cell development. The 3’KE was used to create a 3’KE/BCL-XL transgenic mouse. It was expected that this transgene would alter B-cell compartment composition, as the B-cells in late developmental stages would be rendered resistant to apoptosis. Indeed, we found significant increases in these cell populations, and nests of PC’s were found in the bone marrow of 3’KE/BCL- XL mice. Additionally, perivascular foci of PC’s with nuclear atypia occur in multiple soft tissues in aged 3’KE/BCL-XL mice, and other sequelae consistent with excess Ig production are common. To accelerate B-lymphoid malignancies, we crossed the 3’KE/BCL-XL mouse to an E/MYC or E/MYC transgenic mouse (these mice use the IgH E or E enhancers to drive MYC expression). While Eµ activity begins early in B-cell development and continues throughout all other developmental stages, Eα activity is developmentally restricted to late stages of development and is especially influential in governing heavy chain Ig expression in PC’s. Co-expression of BCL-XL and MYC under the context of the 3’KE and the E leads to a highly fatal B-lymphoid malignancy with a median survival of 5.5 weeks. When MYC expression is controlled by the E, however, fatal PC neoplasms in the bone marrow and other lymphoid organs develop with a median survival of 14.5 weeks. To facilitate visualization of normal and malignant PC’s, we are employing the use of the PC-GFP mouse, which uses kappa Ig regulatory elements to drive GFP expression in PC’s. To investigate the role of constitutively activated RAS expression in B-lymphoid malignancy, we have generated a 3’KE/N-RASV12 mouse. To elicit antigen-induced clonal expansion of the genetically altered plasma cells, we are immunizing the genetically engineered mice. In summary, deregulating apoptotic and oncogenic pathways in plasma and other B-cells by using novel transcriptional regulators in genetically engineered mice serves as a good platform to further the understanding of B- lymphoid malignancies, including MM. S183
216 Transgenic expression of Myc and IL-6 in B cells causes plasma cell tumor formation in mice Siegfried Janz 1, Joong Su Kim 1, Alexander L. Kovalchuk 1, Seong Su Han 1, Sung Sup Park 1, Lino Tessarollo 2, Nicole McNeil 3, Thomas Ried 3, Ted A. Torrey 4, Herbert C.Morse III 4 and Michael Potter 1 1 Laboratory of Genetics, NCI, 2 Mouse Cancer Genetics Program, NCI, 3 Laboratory of Immunopathology, NIAID, 4 Genetics Branch, NCI, NIH, Bethesda. Purpose: Accurate mouse models of human plasma cell tumors are needed to study the events that are involved in the initiation and progression of these neoplasms and to test new intervention strategies that might lead to a better outcome. Based on the fact that the cellular oncogene, MYC, and the B-cell growth, differentiation and survival factor, IL-6, are key players in the pathogenesis of human plasma cell tumors including multiple myeloma, we decided to study plasma cell tumor development in mice that express a human IL-6 transgene and/or a human MYC or mouse Myc transgene in B cells. Mouse models: Three different model systems were utilized to study IL-6 and MYC/Myc driven plasma cell tumor formation in mice. The first model is the BALB/c.H2-Ld-IL-6 congenic strain that harbors a human IL-6 transgene controlled by the widely expressed H2-Ld promoter. The second model is a doubly transgenic mouse that carries in addition to the IL-6 transgene of the first model a human MYC gene driven by the human Igλ enhancer (λ-MYC mice). The third model is a gene-targeted mouse, designated IgH-MycE, which was generated by inserting a histidine-tagged mouse Myc gene, MycHis, into the mouse Ig heavy-chain locus, IgH, just 5' of the intronic enhancer, Eµ Results: All three transgenic mouse strains are characterized by the spontaneous development of plasmacytomas in extramedullary lymphoid tissues with secondary involvement of the bone marrow. Approximately 50% of BALB/c.H2-Ld-IL-6 mice developed IgG plasmacytomas in lymph nodes and spleen by 18 months of age. Virtually all tumors contained Mycactivating chromosomal translocations. Bone marrow infiltration with malignant plasma cells occurred at a late stage of tumor development (plasma cell leukemia). Doubly transgenic λ- MYC/IL-6 mice developed plasmacytomas in the gut-associated lymphoid tissue, often beginning in Peyer’s patches. Bone marrow involvement was variable. The mature plasmacytic phenotype of the λ-MYC/IL-6 tumors was apparently caused by the constitutive expression of IL-6, as singly transgenic λ-MYC mice (without the IL-6 transgene) developed lymphoblastic B- cell lymphoma resembling human Burkitt lymphoma. Approximately 20% of the gene-inserted IgHMycEµ mice developed spontaneous lymph node plasmacytomas by 21 months of age. Bone marrow involvement was common and often widespread. Conclusions: Our findings demonstrate that IL-6 and Myc are crucial for plasma cell tumor formation in mice. The newly developed mice are useful for studying the mechanisms by which IL-6 and Myc promote neoplastic plasma cell development. In addition, they afford a valuable pre-clinical model system in which pharmacological approaches to inhibiting plasma cell neoplasia by interrupting IL-6 signaling or deregulated Myc expression can be tested. 217 TAILOR-MADE MOUSE MODELS FOR STUDYING HUMAN MULTIPLE MYELOMA. María Pérez-Caro, Manuel Sánchez-Martín, Inés González- Herrero, Isidro Sánchez-García Laboratory 13, Instituto Biología Molecular y Celular del Cáncer (IBMCC), Centro de Investigación del Cáncer (CIC), CSIC/University of Salamanca, Campus Unamuno s/n 37007- Salamanca SPAIN In the last few years there have been remarkable advances in our understanding of the molecular biology of human multiple myeloma (MM). This advance has been particularly evident in the case of gene activation by MM-specific chromosomal abnormalities. However, it is still not clear how these chromosomal abnormalities cause the clinical MM features. We believe that this is in part due to the fact that most studies have evaluated the effect of these genes in cell line models, but not in proper animal models. We are mimicking MM in the mouse by modelling the rearrangements of the IgH locus with various partner genes (CCND1, FGFR3, c-maf) associated with human MM. The strengh/novelty of this approach is that the MMchromosomal abnormalities will be generated in embryonic stem (ES) cell-derived mice by a strategy involving sequential gene targeting and Cre/loxP site-specific recombination using methods develop in our lab to assess the effect of chromosomal abnormalities in cancer development. This will involve targeting loxP sites to appropiate positions within the genes involved in the translocations and then using Cre recombinase to catalyze recombination between them to generate the translocation in the mice. The translocation will be activated in a cell-specific manner by expressing Cre recombinase under control of lymphoid specific promoters. These novel mouse models will serve as relevant models to examine the pathophysiology of MM and to develop new therapeutic strategies. 218 Disruption of the Major Vault Protein (MVP, LRP) strongly influences VPARP, TEP1 and vRNA levels, but does not induce hypersensitivity to cytostatics. Marieke H. Mossink1, Arend van Zon1, Erna Fränzel- Luiten1, Martijn Schoester1, George L. Scheffer2, Rik J. Scheper2, Erik A. C. Wiemer1 ,Pieter Sonneveld1 1Department of Hematology, Erasmus University Rotterdam, Netherlands 2Department of Pathology, Academic Hospital "Vrije Universiteit" Amsterdam, the Netherlands. Vaults are large ribonucleoprotein complexes with a distinct structure and a high degree of conservation between species. The cellular localization of the complex, mostly in the cytoplasm but with a vast amount of particles associated with the nuclear suggest a function in cytoplasmic-nuclear shuttling. Also the shape of the complex, a very highly conserved hollow barrel-like structure3 suggests a role in cellular transport. This transport might involve steroid hormones, ribosomes and mRNA . Many reports point to a role of vaults in cellular defense. An increased expression levels of vaults was found in tissues often exposed to xenobiotic elements and in drug resistant cell lines. The expression of the major vault protein (MVP or sometimes referred to as Lung resistance Related Protein, LRP) was associated with a multidrug resistant (MDR) phenotype in human cancer cell lines and in Multiple Myeloma. Several clinical studies indicate that expression of MVP is associated with a poor response to Melphalan in Multiple Myeloma. Gene array analysis has revealed a high expression of LRP in Multiple Myeloma. S184
Page 1 and 2:
Focussed Symposia Arsenic trioxide
Page 3 and 4:
assess whether such a program will
Page 5 and 6:
The overall response rate was noted
Page 7 and 8:
Figure 5A Table 1: VEL + THAL for R
Page 9 and 10:
naturally occurring OPG. Present tr
Page 11 and 12:
0.505). Finally, the multiple event
Page 13 and 14:
CLINICOPATHOLOGICAL DEFINITION OF W
Page 15 and 16:
Plenary Sessions 1. Development of
Page 17 and 18:
clonotypic IgH VDJ signature confir
Page 19 and 20:
can be considered as two entities,
Page 21 and 22:
immunofluorescence staining for DKK
Page 23 and 24:
P2.3 DEVELOPMENT OF A MULTIPLE MYEL
Page 25 and 26:
P2.5 MOLECULAR CYTOGENETICS OF MYEL
Page 27 and 28:
clear that the biggest challenge fo
Page 29 and 30:
esponse and have demonstrated that
Page 31 and 32:
variable region of the idiotypic im
Page 33 and 34:
4. From MGUS to symptomatic MM Fig.
Page 35 and 36:
(MRI); some have claimed that level
Page 37 and 38:
P4.5 CYTOKINES IN MGUS: THERAPEUTIC
Page 39 and 40:
preventing phosphorylation of p70S6
Page 41 and 42:
transducing component of the interl
Page 43 and 44:
survive initial drug exposure and a
Page 45 and 46:
quiescent counterpart, the human um
Page 47 and 48:
transcriptional activity of NF-êB;
Page 49 and 50:
manifestations and anomalous protei
Page 51 and 52:
P7.4 ROLE OF SERUM FREE LIGHT CHAIN
Page 53 and 54:
P7.6 IMMUNOPHENOTYPIC INVESTIGATION
Page 55 and 56:
presumably through the circulation,
Page 57 and 58:
9. Chemotherapy, maintenance treatm
Page 59 and 60:
stratified according to their antim
Page 61 and 62:
explore higher doses of zoledronic
Page 63 and 64:
initial therapy. However, the role
Page 65 and 66:
P10.2.2 SINGLE VERSUS TANDEM HIGH D
Page 67 and 68:
With a median follow-up of 38 month
Page 69 and 70:
cells could be harvested following
Page 71 and 72:
11. What is the role of allogeneic
Page 73 and 74:
found that 75% of patients who were
Page 75 and 76:
patients with responsive disease 3
Page 77 and 78:
9. Giralt S, Estey E, Albitar M, et
Page 79 and 80:
Badros A, Barlogie B, Siegel E, et
Page 81 and 82:
Figure 3b. Event-free Survival 4-Ye
Page 83 and 84:
improve patient outcome in MM. Impo
Page 85 and 86:
myeloma and in 40% of previously un
Page 87 and 88:
degrade OPG in the same way as myel
Page 89 and 90:
P12.2.5 MONOCLONAL ANTIBODY THERAPY
Page 91 and 92:
myeloma. Blood 2001; 98:210-216. 6.
Page 93 and 94:
MHC molecules, in effectively prese
Page 95 and 96:
mice demonstrate that class II-spec
Page 97 and 98:
detected in 293 and NIH3T3 cells. S
Page 99 and 100:
hypothesis that (1) CCND1 and FGFR3
Page 101 and 102:
patient samples. In addition, the p
Page 103 and 104:
016 NEUTRAL ENDOPEPTIDASE (CD10) KN
Page 105 and 106:
2. Genetic heterogeneity in MM: imp
Page 107 and 108:
evidence from two t(11;14) cases wh
Page 109 and 110:
immunoglobulin heavy chain (IgH) lo
Page 111 and 112:
034 Instability of Pericentromeric
Page 113 and 114:
clusters were found, the first was
Page 115 and 116:
MGUS but most likely not crucial fo
Page 117 and 118:
Objective: To compare the impact on
Page 119 and 120:
4 patients had MMSET/IgH but did no
Page 121 and 122:
and clonal PC from MGUS, MM and PCL
Page 123 and 124:
059 VEGF receptor expresion in Mult
Page 125 and 126:
two HLA-A2+ myeloma patients with C
Page 127 and 128:
molecules expressed on the surface
Page 129 and 130:
Recognition of these antigens appea
Page 131 and 132:
Diagnosis requires a single area of
Page 133 and 134:
081 Incidence of solid tumors in co
Page 135 and 136:
Growth Factor (VEGF), Hepatocyte Gr
Page 137 and 138:
PC-AI levels of MGUS and SMM patien
Page 139 and 140:
093 The predominant pathway of tran
Page 141 and 142: was associated with I.V. line, whil
Page 143 and 144: 103 Vascular amyloidosis induces se
Page 145 and 146: 5. Signal transduction pathways and
Page 147 and 148: integrin in IGF-1-triggered MM cell
Page 149 and 150: 118 IL-6- Induced Phosphorylation o
Page 151 and 152: 123 THE IGF/IGF-1R SYSTEM IS A MAJO
Page 153 and 154: human myeloma cell line (HMCL) to a
Page 155 and 156: ligation or dexamethasone (Georgii-
Page 157 and 158: 6. Role of microenvironment 6.1 Cel
Page 159 and 160: 141 Human myeloma cells adhere to f
Page 161 and 162: 145 STUDY OF BONE MARROW ANGIOGENES
Page 163 and 164: patients, the growth of primary mye
Page 165 and 166: tibia (con=49.1±0.7mg/cm2 vs 5T2=4
Page 167 and 168: 157 The Nitrogen-Containing Bisphos
Page 169 and 170: 7. New prognostic criteria for clas
Page 171 and 172: atio of >3. This system has subdivi
Page 173 and 174: Patient 1 (IgG/κ);IgG - 16.5 days,
Page 175 and 176: 176 Pro-inflammatory and angiogenic
Page 177 and 178: 180 Clinical study on the bone lesi
Page 179 and 180: 7.2 Imaging studies 185 99mTc-MIBI
Page 181 and 182: 189 Factors Predicting Occult Spina
Page 183 and 184: vivo detected resonances was invest
Page 185 and 186: Support Unit (CTSU) with flexibilit
Page 187 and 188: (β2m) & C reactive protein (CRP).
Page 189 and 190: plasmids carrying the clonotypic in
Page 191: newly diagnosed multiple myeloma as
Page 195 and 196: 221 Prolonged survival in the 5T2MM
Page 197 and 198: 9. Chemotherapy, maintenance, treat
Page 199 and 200: 229 EFFECTIVENESS OF STANDARD CHEMO
Page 201 and 202: 234 Melphalan and Dexamethasone- Is
Page 203 and 204: Methods. We investigated the VECD p
Page 205 and 206: 9.2 Renal complications. 243 MERIT
Page 207 and 208: cost of SRE-related care was $10,24
Page 209 and 210: those with Hb >13 g/dL. Analysis of
Page 211 and 212: sustained response, lasting from 52
Page 213 and 214: proportion of bone abnormality. IgD
Page 215 and 216: disease. The lack of response to in
Page 217 and 218: yielding a median of 3x106/kg CD34
Page 219 and 220: patients(pts) aged ≥60 yrs. We co
Page 221 and 222: PBSC mobilizing regimen utilizing C
Page 223 and 224: their leukocytes and platelets. No
Page 225 and 226: 25 Gy to marrow. The tracer dose wa
Page 227 and 228: non-CR after the first transplant a
Page 229 and 230: 296 PERIPHERAL BLOOD STEM CELL TRAN
Page 231 and 232: References Effect of dose-intensive
Page 233 and 234: with cyclosporine A and methotrexat
Page 235 and 236: 11.Role of novel therapies targetin
Page 237 and 238: 312 Thalidomide as Rescue of Relaps
Page 239 and 240: patients, light chain in 1 patients
Page 241 and 242: platelets of 207 (76-402), B2M of 3
Page 243 and 244:
325 Low Dose Thalidomide plus Dexam
Page 245 and 246:
in 5 pts (11%), M protein decreased
Page 247 and 248:
time from diagnosis was 3.7 years a
Page 249 and 250:
339 Thalidomide, Clarithromycin and
Page 251 and 252:
344 COMBINATION OF THALIDOMIDE, DEX
Page 253 and 254:
experienced early death during the
Page 255 and 256:
untreated patients with high tumor
Page 257 and 258:
357 Thalidomide protects endothelia
Page 259 and 260:
362 EOSINOPHILIA IS A VERY COMMON F
Page 261 and 262:
11.5 CC 4047, PS 341 and arsenic tr
Page 263 and 264:
without chromosome 13. Mean IC50 fo
Page 265 and 266:
myeloma patients. Phase I data indi
Page 267 and 268:
11.6 Other drugs 380 EFFECT OF STI5
Page 269 and 270:
significant inhibition of cell prol
Page 271 and 272:
which acts to prevent the synthesis
Page 273 and 274:
of B and its metabolites, however,
Page 275 and 276:
and 10 months after start of therap
Page 277 and 278:
terminal kinase (JNK) pathway which
Page 279 and 280:
lymphocytes was tested by Ellispot.
Page 281 and 282:
411 Dendritic Cell-Based Idiotype V
Page 283 and 284:
Author Index Abe M 74 160 Abelman W
Page 285 and 286:
De La Serna J 291 De Laurenzi A P11
Page 287 and 288:
Hull DR 338 Hullen C P10.2.1 Humes
Page 289 and 290:
Morra E 249 280 359 Morris CM 226 M
Page 291 and 292:
Siegel D 70 115 250 Sierra J 304 30
show all

Haematologica 2003 - Supplements

Create successful ePaper yourself

Delete template?

Save as template?