bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of a Diagnostic Gene Signature for SGN-40, Anti-CD40 Monoclonal Antibody, in Pre-Clinical NHL Models and the Role of FAS in SGN-40 Mediated Apoptosis.
Authors: Shi Xiaoyan , Burington Bart , Januario Tom , Lau Jeffrey T , Yu Shang-Fan , Ebens Allen , Dornan David 
Published: April 2020

Abstract
SGN-40, an anti-CD40 monoclonal antibody, is a humanized IgG1 antibody that binds to CD40, mediates effector cell functions (ADCC/ADCP) and activates downstream signaling pathways. SGN-40 has shown activity in a phase I single agent multi-dose trial in non Hodgkin’s lymphoma, with greatest activity in diffuse large B-cell lymphoma (Advani et al. 2008, ICML). Previous in vitro studies implicated down-regulation of the germinal center expressed protein Bcl-6, upregulation of p53 family member TAp63a, and FAS death receptor induction as potential mechanisms leading to lymphoma cell death (Lewis et al. 2007, AACR, ASH). In order to further define the apoptosis signaling mechanism, we assessed the ability of SGN-40 to inhibit proliferation and promote apoptosis across a large panel of non-Hodgkin’s lymphoma cell lines. SGN-40 reduced cell viability in 58% (18/31) of cell lines tested. To identify the genes that may be promoting apoptosis and/or inhibiting proliferation, we compared gene expression levels before and after SGN-40 exposure in both sensitive and resistant cell lines, as well as in normal B-cells. SGN-40 strikingly and specifically upregulated FAS on the cell surface of sensitive cell lines. Furthermore, the addition of soluble FAS-Fc dampened SGN-40-induced apoptosis in a subset of sensitive cell lines, suggesting a dependence on a FAS-FASL interaction. These data imply that FAS-dependent apoptosis may directly contribute to the anti-tumor effect of SGN-40. Our data also demonstrate that SGN-40 sensitivity is dependent on the point in B-cell development at which the NHL cell lines were transformed. Sensitive cell lines had a gene signature characteristic of minimal activation of CD40 signaling prior to SGN-40 exposure, whereas resistant cell lines had a signature consistent with prior constitutive signaling downstream of CD40. Thus, SGN-40 appears to elicit its apoptotic properties through activation of CD40 signaling in NHL cell lines not previously exposed to CD40L signaling in the germinal center environment at the time of lymphocyte transformation (GCB lymphomas). In order to develop a clinically feasible assay from FFPE tissue, we developed a 14-gene signature by Stepwise Linear Modeling, utilizing genes from the CD40 pathway activation and GCB gene sets; the classifier gave &gt;96% accuracy (30/31) on the ‘training’ set of cell lines and 75% (3/4) accuracy on a ‘test’ set of xenografts. Overall, our data provides unique insights into SGN-40 mechanisms of action, provides a testable hypothesis of the clinical mechanism of action, and a potential diagnostic test to identify patients more likely to benefit from SGN-40. Efforts are currently underway to test the clinical relevance of these findings.
The Heterogeneous Prognosis of Patients with Myelodysplastic Syndrome (MDS) and Chromosome 5 Abnormalities: How Does It Relate to the Original Lenalidomide Experience in MDS?.
Authors: Kantarjian Hagop M , O’Brien Susan , Ravandi Farhad , Borthakur Gautam , Faderl Stefan , Bueso-Ramos Carlos , Abruzzo Lynne , Pierce Sherry , Shan Jianqin , Issa Jean-Pierre , Garcia-Manero Guillermo 
Published: April 2020

Abstract
Purpose. The positive lenalidomide experience in lower risk myelodysplastic syndrome (MDS) with deletion 5q and red blood cells transfusion dependence resulted in its broad use in MDS. The study aim was to define the prognosis in MDS and deletion 5q with or without other cytogenetic abnormalities.
Patients and Methods. Patients with MDS (&lt;20% blasts) and evaluable cytogenetic studies (1966-present) were reviewed. Outcome of patients with deletion 5q or with loss of chromosome 5 (monosomy 5) was evaluated by the presence or absence of additional chromosomal abnormalities, overall, and in “lower risk” MDS.
Results. Of 2743 patients analyzed, 287 (10%) had deletion 5q, and 216 (8%) had monosomy 5 abnormalities. The median survival was 9 months with deletion 5q, 6 months with monosomy 5, and 17 months without a chromosome 5 abnormality. Considering patients with deletion 5q, the median survival was 33 months with deletion 5q alone, 17 months with deletion 5q and one additional abnormality, but only 6–12 months with deletion 5q and ≥ 2 abnormalities or chromosome 7 abnormality. Only 93 patients (3.4% of total) had lower risk MDS (International Prognostic Scoring System low or intermediate 1) and deletion 5q; their median survival 29 months (2-year survival 60%), and ranged from 13 to 41 months depending on the presence or absence of additional chromosomal abnormalities. Among 58 patients with lower risk MDS deletion 5q with identical entry criteria to the original lenalidomide study, the median survival was 44 months and the 2-year survival rate 80%. Among 198 patients with MDS with deletion 5q and &lt; 10% blasts (a subset now often offered lenalidomide) the median survival was 12 months. Monosomy 5 was significantly more frequently associated with advanced MDS and with other chromosomal abnormalities. Considering only patients with IPSS intermediate 2 or high risk, the survival was better with deletion 5q versus monosomy 5 (medians 7 versus 6 months; p = 0.005).
Conclusions. The prognosis of MDS with chromosome 5 abnormalities is heterogeneous. This study provides baseline expectations of outcome in different MDS subsets and deletion 5q, which could be used for comparative purposes with the lenalidomide experience.
Bortezomib Targets Stroma-Mediated APO2L/TRAIL Apoptosis Resistance in Multiple Myeloma.
Authors: Perez Lia , Parquet Nancy , Anasetti Claudio , Dalton William 
Published: April 2020

Abstract
TRAIL mediates graft versus tumor responses while sparing normal tissues. Previous studies in our laboratory showed that myeloma:stroma microenvironment interactions confer resistance to TRAIL-induced apoptosis, a process referred to as environmental mediated-immune resistance (EM-IR) by increasing synthesis of the anti-apoptotic protein c-FLIP (J Immunol, 180: 3; 2008). Since bortezomib influences TRAIL signaling proteins and targets stroma induced survival signals, we studied whether it reverses TRAIL EM-IR. Myeloma cells (RPMI 8226 and U266) were treated either adhered to HS5 stroma cells or exposed to HS5-released soluble factors in a transwell system with myeloma cells in the upper well and HS-5 stroma cells in the lower well. Pretreatment with bortezomib (10nM for 20 hours) not only induced synergistic cytotoxicity in myeloma but did overcome TRAIL (10mg/mL for 4 hours) apoptosis resistance induced by HS5-released soluble factors. Mechanistic studies showed that bortezomib is not cytotoxic to HS5 stroma cells and does not alter HS5 release of IL-6, a cytokine known to mediate APO2L/TRAIL EM-IR (J Immunol, 180: 3; 2008). Cytokine array testing showed that stroma-myeloma co-culture lead to a significant increase in IL-10 levels but bortezomib had no effect on IL-10. Bortezomib only decreased MCP-2 and MCP-3 levels without affecting other inflammatory cytokines tested. Expression of DR5 receptor (agonistic) and DCR1-DCR2 receptors (antagonist) were increased at 20 hours bortezomib treatment. Pretreatment with DR4/DR5 blocking antibodies suggested that TRAIL signals trough DR4, not DR5, in these cells and increased of decoy receptors suggest that receptor expression could not predict TRAIL sensitivity. Among the proteins involved in TRAIL signaling that are bortezomib targets, HS5-released factors increased only c-FLIP expression, without affecting Heat Shock proteins, BcL-2 and/or IAP protein families. In the presence of HS5 stroma cells, bortezomib reduced c-FLIP thereby contributing to overcome apoptosis resistance. Since c-FLIP is a NF-κB target gene, we studied whether stroma cells lead to NF-κB activation. Gel shift assay showed increased NF-κB activation in myeloma cells treated with HS5 stroma. Bortezomib decreased NF-κB activation in myeloma cells in suspension, but was not effective in the presence of stroma. Since bortezomib abrogates cytokine-induce kinase activation and understanding that c-FLIP may be under control of these kinases, we next studied the effect of stroma and/or bortezomib on proteins involved in IL-6 signaling. HS5-released factor(s) induced STAT-3 and ERK phosphorylation but not AKT activation, although bortezomib effects on c-FLIP shown to be independent of these pathways. Our findings provide the rationale to combine bortezomib to APO2L/TRAIL treatment to abrogate the positive influence of the stroma microenvironment on myeloma cell survival.
Genomic Complexity and Recurrent Aberration in Multiple Myeloma: Analysis of a Compendium of aCGH and Gene Expression Profiles.
Authors: Chng Wee-Joo , Lee Ian , Jimenez-zepeda Victor , Braggio Esteban , Keats Jonathan , Bergsagel Peter Leif , Fonseca Rafael 
Published: April 2020

Abstract
Multiple Myeloma (MM) is the second most prevalent haematological disorder in the US with great recent progress propelled by various technological and therapeutic advances. Nevertheless, the complexity of MM genomes remains inadequately characterized. We have combined high-resolution (44K Agilent) array comparative genomic hybridization (aCGH) and gene expression data with clinical outcomes in an initial attempt at indexing genomic complexity in MM and its clinical implication in 100 MM patients. We enumerated the Number of Aberrations per Tumor (NAPT) as defined by the ADM-1 algorithm available within CGH Analytics. There appear to be no difference in degree of genetic complexity between hyperdiploid and non-hyperdiploid myeloma. Generally, t(11;14) have low NAPT whereas D2 tumors have high NAPT. Amongst the high-risk genetic subtypes, tumors with maf translocations have relatively low NAPT whereas t(4;14) have high NAPT. Whilst a Multivariate Cox Proportional Hazards regression of survival outcome showed little statistical support for popular indicators such as ploidy and TC classification, results for NAPT were considerably more encouraging (p~ 0.07, median survival of 40 months in low complexity vs 18 months in high). Log-rank tests further supported prognosis based on NAPT independent of clinical status and ploidy. To characterize complexity, we clustered aCGH profiles by clinical status, ploidy and TC classification. Clear patterns of aggregation and recurrent aberration were observed suggesting that they were possibly instrument to aberrant gene expression. Indeed, expression of a large proportion of genes approximately 2Mb of the recurrent breakpoints varied significantly between patients carrying aberrations vis-a-viz others that did not. Comparison against known gene sets showed an enrichment for cell-cycle and apoptosis genes. We are presently characterizing several such breakpoint-associated genes for their functional significance.
A Pre-Existing Plasma Cell Disorder Occurs in Most Patients with Multiple Myeloma.
Authors: Weiss Brendan M , Verma Pramvir , Abadie Jude , Howard Robin , Kuehl Michael 
Published: April 2020

Abstract
Background. A pre-existing plasma cell disorder (PPCD), such as monoclonal gammopathy of undetermined significance (MGUS), is thought to be present in at least one-third of patients presenting with symptomatic multiple myeloma (MM). However, no study has comprehensively evaluated the proportion of patients with MM that had a PPCD by laboratory testing on pre-diagnostic sera.
Methods. The Walter Reed Army Medical Center autologous stem cell transplant database was cross-referenced with the Department of Defense Serum Repository (DoDSR) database, which catalogs serum samples collected every 2 years on over 4 million active-duty service members. All samples 32 years prior to the diagnosis of MM were retrieved. Serum protein electrophoresis (SPEP), immunofixation electrophoresis (IFE) and serum free light chain analysis (sFLC) (The Binding Site, San Diego, CA) were performed on all samples. A PPCD was defined as a positive SPEP, IFE or abnormal sFLC ratio.
Results. Serum samples prior to the diagnosis of MM were available for 30/90 patients, and the median number of samples per patient was 3.5 (range, 1–14). The median age at diagnosis of MM was 48.1 yrs (29–67), with 96% male, 53% Caucasian, and 47% African-American. The Ig isotype of MM was IgG 76%, IgD 10%, light-chain 7%, and non-secretory 7%. A PPCD was detected in 27/30 patients (90%, 95% CI 74–97%). The initial PPCD was detected by sFLC alone in 6/27 (22.2%), IFE alone 2/27 (7.4%), SPEP+IFE 5/27 (18.5%), SPEP+IFE+sFLC 13/27 (48.1%) and IFE+sFLC 1/27 (3.7%). There were 4 patients whose only positive sera was 2.5–3.5 years prior to diagnosis, with all preceding sera negative.
Conclusions. First, a pre-existing plasma cell disorder is present in most MM patients at least 2.5 years prior to diagnosis. Second, consistent with published evidence for a small fraction of patients with high risk MGUS, 4/30 patients were documented to progress rapidly through an MGUS phase to MM. Third, 4/4 patients with light chain only or non-secretory MM had a PPCD that was detected only by sFLC, thereby indicating that all these tumors are preceded by a light chain only or non-secretory PPCD.
Decreased Thymosin Beta 4 Expression Results in Poor Prognosis and Decreased Survival in Multiple Myeloma.
Authors: Caers Jo , Hose Dirk , Kuipers Ine , Bos Tomas Jan , Van Valckenborgh Els , Menu Eline , De Bruyne Elke , Goldschmidt Hartmut , Van Camp Ben , Klein Bernard , Vanderkerken Karin 
Published: April 2020

Abstract
Thymosin beta 4 (Tβ4) is a small cytoplasmic and nuclear protein involved in different cellular processes including differentiation, migration and proliferation. It is overexpressed in several malignant cells, resulting in an increased angiogenic respons and metastatic potential of tumor cells. In the current study we wanted to evaluate the expression of Tβ4 in primary human multiple myeloma cells (MM) and murine 5TMM cells. Microarray analysis and qRT-PCR of 171 MM patients treated with high dose therapy and autologous stem cell transplantation, showed that MM cells display a decreased Tβ4 expression compared to plasma cells from healthy individuals. We investigated whether Tβ4 expression in MM cells influences the event free (EFS) and overall survival (OAS) of these patients. As Tβ4 was expressed in all MM cells, we compared survival of patients with Tβ4 expression in MM cells in the upper thirtile (Tβ4high MMC) against the lowest thirtile (Tβ4low MMC). Patients with Tβ4high MMC had an increased EFS (P&lt; 0.05) and also their OAS tended to be longer. A similar observation was made in the 5TMM model where qRT-PCR and ELISA revealed a decreased expression of Tβ4 in BM samples from 5T33MM and 5T2MM diseased mice compared to control mice. To study the functionality of Tβ4, we overexpressed Tβ4 in 5T33MMvitro (5T33MMvt) cells by lentiviral transduction. These 5T33MMvt-Tβ4+ cells demonstrated a significantly decreased proliferative capacitity and an increased sensitivity to different anti-myeloma agents (bortezomib, melphalan, dexamethasone) compared to control 5T33MMvt cells. Furthermore, injection of 5T33MMvt-Tβ4+ cells in C57BlKaLwRij mice resulted in a significant decreased tumor formation (paraprotein concentration was 1.92 g/dl versus 3.74 g/dl for untransduced cells, p&lt;0.05) and a prolonged survival compared to mice injected with untransduced 5T33MMvt cells (respectively 65.9 days versus 88.9 days, p&lt; 0.05). In conclusion, we found a decreased Tβ4 expression in human and murine MM cells compared to normal plasma cells. In our cohort of MM patients treated with high-dose chemotherapy, low expression of TB4 identifies a group of patients with adverse prognosis while in the murine 5TMM33vt model, overexpression of Tβ4 resulted in an inhibitory effect on tumor formation suggesting a role of Tβ4 as a tumor suppressor gene in MM.
A Multicenter Phase I/II Dose Escalation Study of Lenalidomide in Combination with Melphalan and Dexamethasone in Subjects with Newly-Diagnosed Light-Chain (AL)-Amyloidosis.
Authors: Moreau Philippe , Jaccard Arnaud , Benboubker Lotfi , Royer Bruno , Coiteux Valerie , Bridoux Franck , Harousseau Jean-Luc , Traulle Catherine , Grosbois Bernard , Leblond Veronique , Alakl May , Hermine Olivier , Fermand Jean-Paul 
Published: April 2020

Abstract
A recent prospective randomized trial comparing standard-dose to high-dose melphalan in patients presenting with newly diagnosed AL-amyloidosis showed that oral melphalan-dexamethasone (M-dex) given monthly could be considered the current standard of care, with a median survival of 56 months (Jaccard, N Engl J Med 2007). The use of a combination of lenalidomide and dexamethasone has also been tested in patients with symptomatic AL-amyloidosis. The initial dose of lenalidomide at 25 mg/day was poorly tolerated. However, a 15 mg/day dose regimen was well tolerated and effective, with both hematologic and organ responses (Dispenzieri, Blood 2007 & Sanchorawala, Blood 2007). Combining M-dex with lenalidomide (M-dex-rev) could increase the response rate but the toxicity of this regimen is still unknown. Thus we have initiated a multicenter single-arm open-label phase I/II dose escalation study of lenalidomide administered in combination with M-dex. The primary endpoint was the incidence of dose limiting toxicities (DLT) during the first cycle of lenalidomide at a given dose level in order to determine the maximum tolerated dose (MTD). In addition to melphalan 0.18mg/kg/day from day 1–4 of each 28 day cycle and dexamethasone 40mg/day from day 1– 4 of each 28 day cycle, 3 cohorts of 3 subjects were successively exposed to escalating doses of lenalidomide (5, 10 and 15mg once daily on days 1–21 of a 28 day cycle). DLT was defined using National Cancer Institute (NCI) common toxicity criteria during the first 4 weeks of treatment (one cycle) as the following: at least grade 2 cardiac arrhythmia, at least grade 3 non hematologic toxicity, grade 4 neutropenia lasting &gt;7 days or any other Grade 4 hematologic toxicity, or treatment delay due to toxicity that occurred during the first cycle. No DLT was observed among the first 3 patients treated at 5, 10 and 15 mg lenalidomide/day. 9 additional subjects will be enrolled at 15 mg/day to better define the safety profile and estimate the hematologic and organ response rate. Final results regarding both toxicity and efficacy will be presented during the meeting.
Similar Rate of Thrombosis in Essential Thrombocythemia and Polycythemia Vera Patients after Stratification for JAK2 V617F Allele Burden.
Authors: Carobbio Alessandra , Finazzi Guido , Antonioli Elisabetta , Guglielmelli Paola , Vannucchi Alessandro M. , Delaini Federica , Rambaldi Alessandro , Barbui Tiziano 
Published: April 2020

Abstract
Patients with Essential Thrombocythemia (ET) can be categorized as either JAK2 V617F mutated (V617F+) or wild type (V617F−). Mutated patients display multiple features resembling Polycythemia Vera (PV), with significantly higher hemoglobin level and neutrophil counts, lower platelet count, more pronounced bone marrow erythropoiesis and granulopoiesis and higher tendency to transform in PV. Presence of the mutation and/or allele burden has been variably associated with the rate of vascular complications in ET and PV, but a direct comparison between the two disorders under this respect has not been performed. To tackle this issue, we compared the rate of major thrombosis in 867 ET patients (57% were JAK2 V617F+) with that in 415 PV patients (all V617F+). The median follow-up was 4.9 (0 – 39) and 3.8 (0 – 26) years in ET and PV, respectively. High risk ET patients (age ≥ 60 years and/or previous thrombosis) received Hydroxyurea whereas the vast majority of low-risk remained untreated. PV patients were treated according to the current risk-stratified recommendations. Thrombotic episodes were recorded over time and calculated as rates % per patient/year (pt/yr). After adjusting for age, the thrombosis-free survival curves of JAK2 V617F+ and V617F− ET patients were superimposable until 10 years after the diagnosis, then they diverged so that the actuarial probability of major thrombosis in mutated ET patients reached that of PV (48% vs 55%, test for trend p=0.05). We found that JAK2 V617F+ allele burden measured by real-time quantitative PCR influenced these rates in a comparable way in both ET and PV. Actually, in JAK2 wild type ET (n=376, 43%) the rate was 1.4% pt/yr. In ET patients with JAK2 V617F+ allele burden ranging from 1 to 25% (N=190; 49%) the rate was 1.9 % pt/yr compared to 1.2 in PV patients (N=64, 19%); in the group with 26–50% the rate was 2.0 % pt/yr in ET (N=177; 45%) and 3.0 in PV patients (N=118, 36%); in cases of V617F+ allele burden greater than 50% the rate was 3.8 % pt/yr in ET (N=23; 6%) and 2.9 in PV patients (N=147, 45%). In conclusion, from this retrospective analysis, we conclude that in patients with ET harboring JAK2 V617F mutation the rate of stroke, myocardial infarction and venous thromboembolic complications is similar to that of PV patients and increases in dependence of V617F allele burden, supporting the hypothesis that ET and PV may be viewed as a continuum also in terms of vascular complications
Molecular Mechanisms of JAK2V617F Oncogenic Activation: The JAK2- V617F Mutant Utilizes the SH2 Domain for Constitutive Kinase Activity in the Presence of Unstimulated Heterodimeric Cytokine Receptor.
Authors: Gorantla Sivahari P , Dechow Tobias , Grundler Rebekka , Peschel Christian , Duyster Justus 
Published: April 2020

Abstract
The JAK2-V617F mutation has been reported in the majority of MPDs including PV, ET, and IMF. This mutation leads to the constitutive activation of the JAK2 tyrosine kinase activity and overexpression of JAK2V617F renders hematopoietic cell lines growth factor-independent. However, the molecular mechanism leading to constitutive activation of JAK2V617F is largely unclear and the requirement of homodimeric or heterodimeric cytokine receptors needs to be determined. Here we show that oncogenic JAK2-V617F requires an intact SH2 domain for constitutive kinase activity. To this end we mutated the conserved arginine 426 within the SH2 domain to a lysine. Ba/F3 cells expressing JAK2V617F grew IL-3-independent and showed constitutive activation of JAK2, STAT5, and ERK1/2. In contrast, introduction of the SH2 mutation in JAK2V617F abrogated both transformation as well as constitutive activation of downstream signaling pathways. Accordingly, reconstitution of JAK2 mutants in a JAK2-negative cell line with IL-3R co-expression revealed reduced activation of JAK2 when the SH2 domain was mutated. It has been reported that JAK2 binding to homodimeric type I cytokine receptor may facilitate JAK2V617F-mediated transformation. Interestingly, co-expression of the homodomeric EpoR with SH2 mutated JAK2V617F rescues the phenotype indicating that the SH2 domain is required for JAK2 signaling in the presence of heterodimeric but not homodimeric cytokine receptors. Membrane localization studies showed equal membrane distribution of SH2-mutated and unmutated JAK2-V617F indicating that the SH2 domain mutation does not affect subcellular distribution of JAK2. However, co-IP experiments revealed a possible role for the SH2 domain in the dimerization and transphosphorylation of JAK2. Consequently, reduced transphosphorylation was seen in IL-3R- but not in EpoR-expressing cells. In a BM transplantation model we found that an intact SH2 domain in JAK2V617F was required for the induction of a MPD-like disease. Thus, our results points to an important role of the SH2 domain for the constitutive activation of JAK2V617F in cells expressing heterodimeric cytokine receptors.