bluuebunny/crossref_metadata_2025_split huggingface dataset:
MPT Vs ThaDD in Very Elderly Patients with Multiple Myeloma: a Case-Match Study.
Authors: Offidani Massimo , Bringhen Sara , Corvatta Laura , Petrucci Maria Teresa , Gentili Silvia , Musto Pellegrino , Polloni Claudia , Gentilini Fabiana , Falcone Antonietta , Caravita Tommaso , Liberati Anna Marina , Brunori Marino , Callea Vincenzo , Piro Eugenio , Grasso Mariella , Cangialosi Clotilde , Galieni Piero , Rossini Fausto , Galli Monica , Montanaro Marco , Alesiani Francesco , Stefano Valerio De , Catarini Massimo , Elice Francesca , Zambello Renato , Visani Giuseppe , Griso Luciano , Angelucci Emanuele , Boccadoro Mario , Leoni Pietro , Palumbo Antonio 
Published: October 2019

Abstract
Abstract 1835
Poster Board I-861
Five studies demonstrated the superiority of MPT over MP regimen in elderly patients with MM not eligible for transplantation. In particular, one of these studies, showed this figure in patients aged more than 75 years who represent more than one third of MM patients. Nevertheless, in this latter study 42.5% of patients withdrawn from the MPT protocol because of toxicity. Therefore, there is a wide room of improving these results in this troublesome patient population. Using ThaDD regimen, including liposomal pegylated doxorubicin, we reported low haematological and non-hematological toxicity in elderly and
Unrelated Stem Cell Transplantation After Reduced Intensity Conditioning for Patients with Multiple Myeloma Relapsing After Autologous Transplantation A Prospective Multicenter Phase II Study of the Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation (EBMT).
Authors: Kröger Nicolaus , Shimoni Avichai , Schilling Georgia , Schwerdtfeger Rainer , Bornhäuser Martin , Nagler Arnon , Zander Axel R. , Heinzelmann Marion , Brand Ronald , Gahrton Goesta , Morris Currly , Niederwieser Dietger , de Witte T. M. 
Published: October 2019

Abstract
Abstract 2308
Poster Board II-285

Introduction:
Dose-reduced conditioning followed by allogeneic stem cell transplantation has become a treatment option for patients with multiple myeloma. However, the experience using unrelated donor is limited.

Patients and Methods:
From 2002 to 2007, 49 myeloma patients with relapse to a prior autologous SCT were included in a prospective multicenter trial to determine the efficacy of a reduced melphalan (140 mg/m2)/fludarabine regimen followed by allogeneic SCT from unrelated donors. GvHD prophylaxis consisted of anti-lymphocyte globulin (ATG-Fresenius®), cyclopsorin A and short course of MTX.

Results:
No primary or secondary graft failure was observed and all patients showed leukocyte and platelet engraftment after a median of 15 and 19 days, respectively. Acute graft-versus-host disease (GvHD) grade II to IV occurred in 25% and chronic GvHD in 35% of the patients. Limited GvHD was seen in 29 % and extensive GvHD was seen in 6 % of the patients. Overall response rate at day 100 was 95% including 46% complete remission (CR). Cumulative incidence (CI) of non-relapse mortality at one year was 25% (95% CI: 13-37%) and significantly lower for HLA matched compared to mismatched SCT (10% vs. 53%, p=0.001). During follow-up 22 patients experienced relapse (54 %) resulting in a cumulative incidence of relapse at 1, and 3 years of 27% (95% CI: 14-40%) and 55% (95% CI: 40-70%), respectively. The median time to relapse was 318 days (r: 56 – 861). After a median follow up of 43 months, the estimated 5-year progression-free (PFS) and overall survival (OS) rates were 20% and 26%, respectively and were significantly better for matched in CR at day 100 (41 vs. 7%, p=0.04 and 56 vs. 16%, p=0.02).

Conclusions:
Allogeneic stem cell transplantation from unrelated donors after a reduced intensity regimen is feasible, but an optimal donor selection is mandatory for a low non-relapse mortality. The high relapse incidence remains a major concern should be improved by including posttransplant strategies to upgrade remission status.

Disclosures:
No relevant conflicts of interest to declare.
Chronic Lymphocytic Leukemia with Stereotyped IGHV4-59/IGKV3-20 B Cell Receptors: Another Manifestation of Hepatitis C Virus-Associated B Cell Lymphoproliferation?.
Authors: Kostareli Efterpi , Janus Agnieszka , Gounari Maria , Belessi Chrysoula , Pospisilova Sarka , Oscier David , Foroni Letizia , Francia di Celle Paola , Jurlander Jesper , Rosenquist Richard , Davi Frederic , Thompson Keith , Ghia Paolo , Stamatopoulos Kostas 
Published: October 2019

Abstract
Abstract 2331
Poster Board II-308
The hepatitis C virus (HCV) has been implicated in the development of B-cell lymphoproliferative disorders, including type II mixed cryoglobulinemia (MC-II) and B-cell lymphoma. MC-II is characterized by the presence of monoclonal IgM autoantibodies with rheumatoid factor (RF) activity. The monoclonal IgMs typically form immune complexes by binding polyclonal IgGs that exhibit anti-HCV reactivity. In a series of 6,196 patients affected by chronic lymphocytic leukemia (CLL), we have identified a subset of 12 cases sharing stereotyped mutated IGHV4-59/IGKV3-20 B cell receptors (BCRs) of the MD isotype (subset #13). Comparison of subset #13 heavy chain sequences to a comprehensive dataset of relevant public-database sequences revealed identical gene usage and remarkable junctional homology with the Ig sequence GenBank/U85234, the heavy chain of a RF detected in a healthy donor, as well as the sequence GenBank/AF303916, the clonotypic heavy chain from a CLL case with a history of HCV-associated MC-II. In addition, the light chain IGKV3-20/IGKJ1 stereotyped rearrangements in subset #13 were closely similar if not identical to the rearrangements expressed by clonally expanded IgM+κ+CD27+ B cells in HCV-associated MC-II. For both heavy and light chains, sequence similarities extended beyond junctional regions to shared, “stereotyped” somatic hypermutations across the entire IGHV and IGKV domain, respectively. We established viable and antibody-secreting heterohybridomas from the leukemic cells of a subset #13 case and confirmed the identity of the produced soluble antibody to the IG expressed by the CLL clone. ELISA tests against various antigens revealed that the soluble stereotyped IGHV4-59/IGKV3-20 antibody exhibited RF activity in vitro, while it was not reactive against HCV antigens. In conclusion, the present study for the first time provides evidence for the potential implication of HCV in the pathogenesis of at least a subset of CLL cases with distinctive stereotyped BCRs. The elucidation of the underlying immune mechanisms may pave the way for tailored anti-viral/anti-leukemic therapy for selected cohorts of patients that can be easily identified by molecular techniques during the diagnostic work-up.

Disclosures:
No relevant conflicts of interest to declare.
High Resolution Array-CGH Provides New Insights Into the Prognosis of Chronic Lymphocytic Leukemia (CLL): Is 8p Loss Worse Than 17p Loss?.
Authors: Rinaldi Andrea , Mian Michael , Rossi Davide , Forconi Francesco , Deambrogi Clara , Spina Valeria , Cencini Emanuele , Sozzi Elisa , Drandi Daniela , Ladetto Marco , Santachiara Rita , Marasca Roberto , Chigrinova Ekaterina , Cavalli Franco M.D. , Gattei Valter , Zucca Emanuele , Gaidano Gianluca , Kwee Ivo , Bertoni Francesco 
Published: October 2019

Abstract
Abstract 2339
Poster Board II-316

BACKGROUND:
CLL, the most common adult-onset leukemia in the Western world, has a heterogeneous clinical course. Many advances have led to a better understanding of its pathogenesis and to improvements in treatment strategies, but striking solutions are still missing. We conducted a study to evaluate the impact of genomic aberrations on the clinical course.

METHODS:
From January 1980 to May 2008, 395 frozen samples of CLL patients, were prospectively collected in four centers. Extracted DNA was analyzed with Affymetrix Human Mapping 6.0 arrays. Normal matched DNA was analyzed for one fourth of the cases. Correlations between minimal common regions (MCR) and clinical parameters were evaluated with the Fisherôs-exact test and their impact on OS with the log-rank test. A p-value after Bonferroni multiple test correction (MTC) (p-adj.) <0.05 was considered as statistically significant. Up to now 266 samples have been analyzed.

RESULTS:
Analysis of the clinical parameters (CPs) and known risk factors (Rai/Binet, age, doubling time, LDH, beta2, IGVH status, p53 mutations, telomere length, CD38, 11q, 17p) was consistent to previous published series. ZAP70 did not affect the clinical course, likely due inter-laboratories variability. After a median follow up of 53 months, 143/239 (60%) of the patients have started therapy and 63/261 (24%) died. 5-yr OS was 82%. Fisher test between the MCRs and CPs revealed an inverse relation between the presence of trisomy 12 by FISH and del13q14.3, an association between del17p and losses of 8p regions and between CD38 and 12q gain. Before MTC, 46 MCRs had a significant impact on OS and 67. After MTC, 3 regions maintained their role: 8p22 loss (38/248, 15%, p-adj.=0.002, median OS: 26 months vs. 48), 17p13.3-11.2 loss (20/248, 8%, p-adj.=0.001; median OS: 10 months vs. 48). In univariate analysis, the log-rank test among pts with 8p-/17p- (8/248, 3%), 8p- (30/248, 12%), 17p- (12/248, 5%), wild type (198/248, 80%) was statistically significant (p<0.001; see figure). Importantly, none of the analyzed clinical and biological parameters was associated with this aberration.

CONCLUSIONS:
Loss of 8p22 designated a CLL subgroup with a worse outcome among all patients and in the subset with 17p loss. Our data suggested that this aberration might constitute an independent prognostic factor to be evaluated in independent studies. Results, including a Cox regression model, will be presented on all 395 cases.

Disclosures:
No relevant conflicts of interest to declare.
Marrow Stromal Cells Induce TCL1 Expression in Chronic Lymphocytic Leukemia B Cells.
Authors: Sivina Mariela , Hartmann Elena , Keating Michael , Wierda William G , Rosenwald Andreas , Burger Jan A. 
Published: October 2019

Abstract
Abstract 2347
Poster Board II-324
The human T cell leukemia/lymphoma 1 (TCL1) oncogene was initially identified as a target of chromosomal translocations and inversions at the 14q32.1 chromosome breakpoint region in T-cell prolymphocytic leukemia (T-PLL). Increased TCL1 expression is seen in follicular lymphoma, Burkitt lymphoma, diffuse large B-cell lymphoma, and chronic lymphocytic leukemia (CLL). Transgenic mice over-expressing TCL1 under control of the mu immunoglobulin gene enhancer develop a CD5+ B cell lymphoproliferative disorder that mimics human CLL, indicating that TCL1 plays a central and/or causal role in the pathogenesis of CLL. However, chromosome aberrations that constitutively activate TCL1 have not (yet) been identified in the vast majority of CLL patients, and therefore the oncogenic mechanism(s) of TCL1 activation in CLL remain unclear. There is growing evidence that external signals from the microenvironment control and regulate the survival and proliferation of CLL cells. Marrow stromal cells (MSC) are highly effective in protecting CLL cells from spontaneous and drug-induced apoptosis, and are used as a model system to study the marrow microenvironment. In order to explore the molecular cross talk between CLL cells and MSC, we co-cultured CLL cells with different MSC and analyzed gene expression changes induced by co-cultures with MSC, an approach similar to our recent study with nurselike cells (Blood 113:3050-8, 2009). For this, RNA was extracted from 19-purified CLL cells from 10 different patients (baseline expression, day 0). Also, the same patients' samples were co-cultured on stroma cells (KUSA-H1, NK-Tert) for 2 and 7 days. At these time points, RNA again was isolated after CD19-purification. Then, gene expression was determined using HG U133 plus 2.0 oligonucleotide arrays from Affymetrix. Gene expression changes were analyzed in individual patients' samples, comparing baseline samples' gene expression to samples after 2 and 7 of co-culture on MSC. We observed relatively homogeneous gene expression changes in CLL cells after co-culture with MSC. We found that TCL1 was among the top 5 genes that were most highly up-regulated by MSC, based on at least 3-fold up-regulation in at least 6 of the paired samples. We also found an up-regulation of TCL1 at the protein level when assessed by immunoblotting and flow cytometry in CLL samples after co-culture with MSC. These findings indicate that MSC can induce and regulate TCL1 expression in CLL, suggesting that the microenvironment plays an even greater role in the pathogenesis of this disease than previously recognized.

Disclosures:
No relevant conflicts of interest to declare.
Molecular Heterogeneity as a Basis for Rational Therapeutics in Chronic Lymphocytic Leukemia.
Authors: Herreros Beatriz , Wang Shaomeng , Bischoff James Robert , Carro Ruben , Córdoba Raul , Delgado Julio , García-Marco Jose Antonio , Gómez Natalia , García Juan F. , Tomáas José Francisco , Martínez-López Joaquín , Rodríguez Antonia , Villuendas Raquel , Piris Miguel A. 
Published: October 2019

Abstract
Abstract 2348
Poster Board II-325
Chronic lymphocytic leukemia (CLL), the most frequent form of adult leukemia in Western countries, is characterized by a highly variable clinical course. Data from our group have shown that the gene expression profile can be used to classify CLL patients into different risk groups, according to a biology-based predictive model that includes genes from essential survival pathways, such as the B cell receptor (BCR) and NF-κB pathways. We propose that this molecular heterogeneity may be associated with variable sensitivity to drugs affecting these survival pathways. Using the Connectivity Map bioinformatic tool, we identified several drugs that may indirectly regulate BCR signaling. We decided to explore one inhibitor of calmodulin (Calmidazolium) further. Based on previously available information about specific signaling cascades, we also selected a PIM kinase inhibitor (C6) and a small molecule inhibitor (SMI) of the BCL2 family (TW-37), which targets Bcl-2, Bcl-XL, and, unlike most SMIs, Mcl-1. An automated drug dispensation protocol coupled to a cell viability assay was set up to test these compounds in primary CLL samples. EC50s were calculated at 72 hours of treatment for each sample and drug. EC50s were in the low μM range for Calmidazolium (0.9-6.8 μM) and C6 (1.5-6.5 μM) and in the nM range for TW-37 (90-740 nM). Patients were stratified using IgVH, SHM, ZAP70, PIM1, PIM2, MCL1 expression, and BCR signature expression as markers. Pharmacodynamic markers such as p-AKT were chosen. Associations of EC50s with SHM and ZAP70 clinical parameters were examined. Strikingly, unmutated cases (4/13), characterized by a worse prognosis, were associated with greater sensitivity (i.e., lower values of EC50) to TW-37 (p = 0.017). Interestingly, ZAP70-positive cases (4/10) also showed a tendency towards greater sensitivity to the TW-37 treatment (p = 0.079). However, variability in the response to Calmidazolium and C6 was not statistically significant, given the small size of the series (11 and 5 samples, respectively). In summary, we show that samples from patients with a poor prognosis (unmutated IgHV and ZAP70-positive) are more sensitive to the BCL2 family SMI, supporting the hypothesis that molecular heterogeneity in CLL is associated with variable drug response.

Disclosures:
Wang: Ascenta Therapeutics: Consultancy, Patents & Royalties, Research Funding. Bischoff:Eli Lilly & Co: Consultancy.
Nucleophosmin Is Required for Macrophage Function and Maturation.
Authors: Sportoletti Paolo , Clohessy John G , Riccardi Luisa , Khanna-Gupta Arati , Chen Jian , Berliner Nancy , Pandolfi Pier Paolo 
Published: October 2019

Abstract
Abstract 238
Nucleophosmin (NPM1) is a multifunctional phosphoprotein predominantly localized in the nucleolus that is involved in numerous cellular processes. The NPM1 gene is frequently a target of genetic alteration in hematological tumors and recent published reports have highlighted the importance of NPM1 in malignancies that affect predominantly the myeloid lineage. Complete inactivation of Npm1 in the mouse disrupts primitive hematopoiesis and results in embryonic lethality. Npm1 heterozygosity produces features similar to those of MDS patients, and can progress to overt leukemia. Here we generated a conditional mousemutant to inactivate Npm1 in the myeloid compartment crossing mice carrying a floxed conditional allele of Npm1 (Npm1fl/fl) with the MRP8-Cre transgenic mouse model. Specific deletion of the Npm1 allele in Npm1fl/fl;MRP8-Cre+ mice resulted in complete loss of Npm1 at the protein level in 40% of peritoneal macrophages analyzed by immunofluorescence. In vitro analysis demonstrates that Npm1 deletion altered peritoneal macrophage functions and influenced monocytic differentiation in Npm1fl/fl;MRP8-Cre+ mice. Specifically we demonstrate that (i) superoxide production was significantly attenuated over time in Npm1fl/fl;MRP8-Cre+ compared with Npm1fl/fl;MRP8-Cre- macrophages; (ii) The chemotactic response was decreased by up to 46% ± 9.6 in Npm1fl/fl;MRP8-Cre+ macrophages when compared to controls; and (iii) Phagocytic index of Npm1fl/fl;MRP8-Cre+ macrophages dropped to 40% of that for the control Npm1fl/fl;MRP8-Cre- macrophages. Analysis of monocyte transcriptional regulators revealed a significant reduction of Maf-b and Irf-8 mRNA expression in Npm1 deficient macrophages, demonstrating an altered maturation process in the absence of Npm1. Our analysis demonstrates that Npm1 loss in the myeloid compartment affects macrophage function and profoundly alters gene expression patterns required for proper monocyte/macrophage development and maturation.

Disclosures:
No relevant conflicts of interest to declare.
Selective and Titratable Effects On AML Genome Wide Methylation Patterning, Transcription and Clonogenicity Using Very Low Concentrations of 5-Aza-2'deoxycytidine.
Authors: Heuston Elisabeth , Farrar Jason E. , Triche Timothy , Buckley Jonathan , Sorensen Poul , Reynolds C. Patrick , Reaman Gregory , Yegnasubramanian Vasan , Arceci Robert J. 
Published: October 2019

Abstract
Abstract 2385
Poster Board II-362
5-Aza-2'deoxycytidine (5AzadC) has significantly contributed to the treatment of myelodysplatic syndromes (MDS) and acute myeloid leukemia (AML). But while the cytotoxic effects of 5AzadC have been well characterized, its influence on methylation-induced cellular reprogramming remains poorly understood. We have treated several AML cell lines at extremely low concentrations of 5AzadC (0 nM to 1.0 nM) over the course of three days, followed by the determination of genome wide methylation changes, alterations in transcription profiles as well as cell viability, proliferation, apoptosis and changes in clonogenicity. The results demonstrate titratable responses on both genomic methylation and transcriptional patterns as well as a selective effect on clonogenicity compared to cytotoxicity. An alternative chemotherapeutic cytosine analog, cytosine arabinofuranoside (AraC), does not show the same selective depletion of clonogenic cells, suggesting that 5AzadC's effects are likely due to altered epigenetic changes associated with cellular reprogramming rather than a direct cytotoxic effect. We are currently evaluating 5AzadC and AraC effects on this population using immunophenotyping methods as well as xenograft models of tumorigenicity. These findings describe a potential role for very low concentrations of 5AzadC in treating acute myeloid leukemia through a selective affect on genome wide methylation patterns leading to altered transcription that differentially effects the clonogenic, leukemic stem cell compartment.

Disclosures:
No relevant conflicts of interest to declare.
AML Survival and In Vivo Progression Is Dependent On β-Catenin Signaling.
Authors: Siapati Elena K , Papadaki Magda , Kozaou Zoi , Rouka Erasmia , Michali Evridiki , Gogos Dimitrios , Savvidou Ioanna , Kyriakou Despoina S , Anagnostopoulos Nikolaos.I. , Vassilopoulos George 
Published: October 2019

Abstract
Abstract 2398
Poster Board II-375
B-catenin is the central effector molecule of the canonical wnt signaling pathway which governs cell fate and differentiation during embryogenesis as well as self-renewal of hematopoietic stem cells. Deregulation of the pathway has been observed in various malignancies including myeloid leukemias where over-expression of β-catenin is an independent adverse prognostic factor. In the present study we examined the functional outcome of stable β-catenin down-regulation through lentivirus-mediated expression of short hairpin RNA (shRNA). Reduction of the β-catenin levels in AML cell lines and patient samples diminished their in vitro proliferation ability without significantly affecting cell viability. In order to study the role of β-catenin in vivo, we transplanted leukemic cell lines with control or reduced levels of β-catenin in NOD/SCID animals and analyzed the engraftment levels in the bone marrow. We observed that while the immediate homing of the cells was not affected by the β-catenin levels, the bone marrow engraftment was directly dependent on its levels. Subsequent examination of bone marrow sections revealed that the reduced engraftment was partly due to the inability of the cells with lower β-catenin levels to dock to the endosteal niches, a finding that was confirmed in competitive repopulation assays with untransduced cells. When we examined the expression levels of adhesion molecules and integrins in engrafted cells in vivo, we observed a significant down-regulation of CD44 expression, a molecule that participates in the interaction of HSCs with the niche. Gene expression analysis of the components of the wnt signaling pathway showed that the pathway is subject to tight transcriptional regulation with minor expression deviations. We did, however, observe an up-regulation in components that participate in the non-canonical wnt signaling pathways such as the WNT5B ligand. Ongoing experiments in normal cord blood CD34+ cells will determine the in vivo role of β-catenin signaling in normal hematopoietic progenitors. In conclusion, our study showed that β-catenin comprises an integral part in the development and progression of AML in vivo, indicating that manipulation of the wnt pathway may hold a therapeutic potential in the management of AML.

Disclosures:
No relevant conflicts of interest to declare.