bluuebunny/crossref_metadata_2025_split huggingface dataset:
Single Agent AMD3100 Mobilization of Peripheral Blood Progenitor Cells for Autologous Transplantation in Patients with Multiple Myeloma (MM).
Authors: Flomenberg Neal , Comenzo Raymond , Badel Karin , Calandra Gary 
Published: October 2019

Abstract
Background: The addition of AMD3100 to a G-CSF mobilization regimen increases the number of hematopoietic stem and progenitor cells (HSPCs) that can be collected by apheresis, allowing the successful (re)mobilization and transplantation of MM patients. This single-arm, open-label, study was therefore initiated to determine if AMD3100 alone could successfully mobilize HSPCs in MM patients for autologous stem cell transplantation (ASCT). This is the first clinical trial of HSPCs mobilized by AMD3100 alone in the autologous transplant setting.
Methods: Patients received 240 μg/kg AMD3100 followed by apheresis 6 hours later. Dosing and apheresis could be repeated for up to 4 consecutive days or until ge;4 × 106 CD34+ cells/kg were collected. Post-transplant follow-up included daily WBC measurements until leukocyte engraftment (≥0.5 × 109/l for 3 consecutive days or ≥1 × 109/l for 1 day), and platelet engraftment (≥20 × 109/l for 7 days).
Results: Nine patients have been enrolled. The median number of CD34+ cells collected was 2.7 × 106/kg (Range:1.9– 4.6), completed in 4 apheresis days by 8 patients and 3 days by 1 patient. Six patients (67 %) mobilized ≥2 × 106 CD34+ cells/kg and 2 pts (22 %) mobilized ≥4 × 106 CD34+ cells/kg. All patients were transplanted; 1 patient received tandem transplants with study cells; 1 patient received tandem transplants, the first transplant with study cells followed by a second transplant with G-CSF mobilized cells. Median time to PMN engraftment was 10 days (Range: 10–11 days); median time to PLT engraftment was 21 days (Range: 15–33 days, except for 1 patient with platelet recovery some time between day 23 and 82). One patient with Plasma Cell Leukemia died of disease relapse at 6 months. Engraftment was durable with a follow-up of over 19 months in the earliest transplanted patient. AMD3100 was safe and generally well tolerated with the most common adverse events being injection site reactions, GI upset, and paresthesias. There were no drug-related severe adverse events.
Conclusions: AMD3100 alone mobilizes autologous HSPCs that provide prompt and durable engraftment, indicating that AMD3100 mobilizes functional HSPCs independent of the presence of G-CSF. These results are consistent with and reinforced by observations made using AMD3100 to mobilize allogeneic HSPCs [Devine et al., ASH 2005, #299]. While the number of cells mobilized by AMD3100 alone is relatively modest, AMD3100 plus G-CSF mobilizes a more primitive subset of CD34+ cells than G-CSF alone which could potentially translate into fewer CD34+ cells being required for transplantation when AMD3100 is part of the mobilization regimen [Seeger et al., ASH 2005, #1969]. Therefore, further evaluation of dose and schedule, as well as the role of CD34+ phenotype, is required to optimize HSPC mobilization with single-agent AMD3100.
An Update on the Role of Thalidomide (THAL) in Total Therapy 2 (TT2) for Newly Diagnosed Patients with Multiple Myeloma (MM): Analysis of Subgroups Defined by Standard Prognostic Factors (SPF) and Gene Expression Profiling (GEP)-Derived Subgroups.
Authors: Shaughnessy John D. , Haessler Jeffrey , Zeldis Jerry , Huang Yongsheng , Zhan Fenghuang , Sawyer Jeffrey , Anaissie Elias , van Rhee Frits , Tricot Guido , Epstein Joshua , Zangari Maurizio , Pineda-Roman Mauricio , Hollmig Klaus , Crowley John , Barlogie Bart 
Published: October 2019

Abstract
Background: THAL, whose activity in MM was discovered in the setting of advanced and refractory disease in the late 1990’s (Singhal, NEJM, 2000), has become the standard front-line therapy in combination with dexamethasone (DEX). In a randomized phase III tandem transplant trial, TT2, a higher complete response (CR) rate and longer event-free survival (EFS) had been observed on the THAL arm (Barlogie, NEJM, 2006). The similar overall survival (OS) on THAL and control arms had been attributed to the routine use of THAL as salvage therapy for the patients randomized to the No-THAL arm and the shorter post-relapse OS among patients randomized to the THAL arm.
Patients and Methods: With a median follow-up on TT2 of 53mo, 107 patients have relapsed and 219 died. Subset analyses were performed to determine whether THAL confers an OS advantage in any subgroup of patients.
Results: 6-yr EFS and OS rates are 48%/63% on THAL and 38%/58% on control arm (p=0.01/0.67). Post-relapse OS is now similar with median durations of 5.3mo/4.3mo among control/THAL arms (p=0.11). According to multivariate analyses of 11 standard prognostic factors, EFS was shorter among patients treated without THAL, in the presence of cytogenetic abnormalities (CA), B2M and LDH elevations and low albumin, whereas CR was favorable; OS was inferior with CA, high LDH, low albumin and in patients not receiving 2nd transplant or not achieving CR. Randomization to THAL was beneficial only in the >2 risk factor group: 6-yr OS was 47% in 31 patients on THAL and 12% in 31 control patients (Figure 1, p=0.01). When examined in the context of GEP (70 gene model-based high versus low risk groups) and inter-phase FISH data (amp1q21), available in 260 patients, the 57 with GEP low risk and absence of amp1q21 receiving THAL had 5-yr OS of 90% compared to 74% among 73 controls (p=0.13).
Conclusion: With longer follow-up of 53mo on TT2, EFS remains superior among patients randomized to THAL; post-relapse survival is no longer inferior among those randomized to THAL; THAL benefited a high-risk subgroup with >2 standard risk factors, whereas no significant `difference has yet emerged among genetically defined subgroups.
Figure Figure
Amplification of 1q21 Is Associated with Poor Outcome after Treatment with Bortezomib in Relapsed/Refractory Multiple Myeloma.
Authors: Drach Johannes , Sagaster Verena , Odelga Victoria , Kaufmann Hannes , Ackermann Jutta , Galhuber Markus , Zojer Niklas , Küenburg Elisabeth , Zielinski Christoph , Wieser Rotraud , Ludwig Heinz 
Published: October 2019

Abstract
Bortezomib is an active agent for treatment of multiple myeloma (MM) and may even be effective in patients (pts) with adverse prognostic factors including unfavorable cytogenetic abnormalities. However, it is unknown whether or not bortezomib may overcome the negative prognostic impact of a CKS1B gene amplification at chromosome 1q21 (amp1q21), which has recently been reported as a negative prognostic factor even in the setting of a total therapy approach. We therefore evaluated chromosome 1q21 among other abnormalities in 46 pts with relapsed/refractory MM who were treated with single-agent bortezomib (1.3 mg/m2 on days 1, 4, 8, and 11 every 3 weeks). Median age of pts was 63 years (range, 40 – 82 ) and median time to bortezomib therapy was 40 months (median number of prior therapies: 3; 96% of pts had high-dose pulsed dexamethasone, 61% thalidomide, 85% alkylating agents, and 41% high-dose melphalan). A response to bortezomib was noted in 45% of pts, with 17% of pts achieving a CR/near CR. Amp1q21 as determined by interphase FISH was observed in 20 of the 46 pts (43.5%). Treatment outcome after bortezomib was negatively affected by presence of amp1q21: The overall response rate was 30% (versus 58% in pts with normal 1q21; P = .06) and the CR/near-CR rate was 10% (versus 23%). Moreover, amp1q21 was associated with shortened time to treatment failure (median, 2.4 versus 6.6 months; P = .043) and overall survival (OS) (median, 4.4 versus 19.8 months; P = .003) compared to pts with normal 1q21. Beta-2-microglobulin and 14q32 translocations were unrelated to treatment outcome with bortezomib. In contrast, median OS was short in the presence of low serum albumin (4.8 versus 17.8 months; P = .036) and deletion of 13q14 (6.7 months versus median not yet reached; P = .06). Using amp1q21, low serum albumin, and deletion of 13q14 as risk factors, patients with significantly different median OS after bortezomib treatment were discriminated: Pts with ≥ 2 risk factors had a median OS of only 4.9 months as opposed to pts with 1 risk factor (median OS 16.5 months) or without any risk factor (median OS not yet reached) (P = .004). In conclusion, FISH-defined amp1q21 is a strong adverse prognostic factor for pts with relapsed/refractory MM treated with single-agent bortezomib. We are currently investigating whether or not bortezomib combinations may be more effective in MM pts with amp1q21.
A Novel Multiplex Drug Screening Assay for Identification of Potential Compounds with Anti-Myeloma Activity from a Library of 1120 Drugs.
Authors: Feng Rentian , Lokshin Anna , Gorelik Elieser , Lentzsch Suzanne 
Published: October 2019

Abstract
The majority of drug screening assays are aimed at selection of compounds that affect proliferation or survival of myeloma cells. However, this approach might fail to identify compounds with a potent therapeutic activity that are unable to directly inhibit tumor cell proliferation in vitro but might have potent anti-tumor activity in vivo by targeting the microenvironment of the myeloma cell. For this purpose we used a Multiplex drug-screening assay (MDSA) to identify compounds with potential anti-myeloma activity from a library of 1120 compounds provided by the Multiple Myeloma Research Foundation (MMRF). MDSA is based on use of the Luminex technology (LabMAP Multianalyte Profiling), and testing various myeloma producing factors (MPFs), such as cytokines, chemokines and growth factors that are important for myeloma cell proliferation and survival. The multiple myeloma cell lines MM1.S, RPMI-8226, and IM9 were tested for their capacity to secrete the full set of 31 cytokines, chemokines and growth factors. RPMI-8226 was selected for MDSA due to its high capacity to secrete MPFs (IL-8, VEGF, MCP-1, MIP-1α, MIP-1β, IP10, RANTES and SIL-6R). RPMI-8226 cells were treated with 10 10−6M of each compound (first screening phase) and 1 10−6M (secondary screening), and supernatants from 72-hour cultures were analyzed. The criterion of effective drugs for each cytokine was set up as the ability to inhibit or stimulate MPFs (exceed +/− 1.5 mean value of non-treated control). The resulting data on the drugs were graded by the degree to which they caused inhibition or stimulation of all MPFs (greater than 50% and greater than 90%).
A total of 205 of the 1,120 candidates were picked out from the first screening at 10 10−6M. Results from the second analysis (at 1 10−6M) indicated that 14 compounds achieved inhibition of all MPFs and dequalinium dichloride manifested the strongest inhibition of all MPFs. Forty drugs were able to selectively inhibit certain MPFs at levels that exceeded 50% and 14 drugs inhibited MPFs by 90%. With respect to stimulation of cytokine secretion, a total of 39 compounds demonstrated selective stimulation of some MPFs and three drugs (amethopterin (R, S), etoposide, and lasalocid sodium salt) induced stimulation at the level of 90% or greater.
Overall, MDSA is a powerful high throughput screening assay to analyze compounds with inhibitory or stimulatory effects on cytokines, chemokines and growth factors that are involved in the pathogenesis of multiple myeloma. Potent compounds identified in this study warrant further investigation for their anti-myeloma effects in vitro and in vivo.
Variation in DNA Repair Genes XRCC3, XRCC4, and XRCC5 and Risk of Myeloma.
Authors: Hayden Patrick , Tewari Prerna , Staines Anthony , Morris Derek , Crowley Dominique , Nieters Alexandra , Becker Nicholas , de Sanjose Silvia , Foretova L. , Maynardie Mark , Coco Pier Luigi , Boefetta Paolo , Brennan Paul , Browne Paul V. , Lawler Mark 
Published: October 2019

Abstract
Aberrant class switch recombination (CSR), the physiological process that regulates maturation of the antibody response, is believed to be an early event in the pathogenesis of myeloma. The genetic basis of CSR, from initiation of the DNA double-strand break through to detection and repair, has been elucidated. We hypothesised that germline polymorphisms in the genes implicated in DNA double strand break repair may contribute to susceptibility to myeloma. We therefore assessed 32 SNPs in 3 genes central to the DNA repair pathway in patients with myeloma and controls from the EpiLymph Study, a European study of the epidemiology of lymphoid neoplasms, and from an Irish hospital registry (306 cases, 263 controls). The genes examined were XRCC3, XRCC4, and XRCC5. XRCC3 is a member of the RecA/Rad51-related protein family that participates in homologous recombination. The XRCC4 protein forms a complex with DNA ligase IV and DNA-dependent protein kinase in the repair of DNA double-strand breaks by non-homologous end joining. XRCC5 encodes the 80-kilodalton subunit of the Ku heterodimer protein, the DNA-binding component of the DNA-dependent protein kinase. SNPs from the chosen genes were identified from HapMap CEU Phase I and II genotype data (Public Release #20; 2006-01-24). Haplotype-tagging SNPs (htSNPs) were chosen based on Tagger analysis (as implemented in Haploview Version 3.2). A SNP in GSTP1 was also genotyped to allow for comparison with allele frequencies previously generated from a myeloma cohort. Genotyping was performed using TaqMan®-based assays on the 7900 ABI HT platform. The drop-out rate was consistently less than 3% in all assays. For quality control (QC) purposes, duplicates of 10% of the samples were interspersed throughout the plates. The concordance rates for QC samples were greater than 98%. GSTP1 SNP results were comparable to previously published findings. For the htSNP rs963248 in XRCC4, Allele A was significantly more frequent in cases than in controls (86.4% vs.80.8%) (p=0.0105), as was the AA genotype (74% vs. 65%) (p=0.026). Haplotype analysis was performed using Cocaphase for rs963248 in combination with additional SNPs in XRCC4. The strongest evidence of association came from the A - T haplotype from rs963248-rs2891980 (80.9% vs. 74.5%; p=0.008). For XRCC5, the genotype GG from rs1051685 was detected in 10 cases from different national populations but in only 1 control (p=0.015). Interestingly, this SNP is located in the 3′ UTR of XRCC5. Overall, these data provide support for the hypothesis that common variation in the genes encoding DNA repair proteins contributes to susceptibility to myeloma.
Histone Deacetylase-6 (HDAC6) Modulates Akt and STAT3 Activity Via Heat Shock Protein (Hsp) 90 in Human Multiple Myeloma (MM) Cells.
Authors: Hideshima Teru , Bradner James E. , Yasui Hiroshi , Raje Noopur , Chauhan Dharminder , Podar Klaus , Tai Yu-Tzu , Mitsiades Constantine , Richardson Paul G. , Mazitschek Ralph , Schreiber Stuart L. , Anderson Kenneth C. 
Published: October 2019

Abstract
Histone deacetylase 6 (HDAC6) has an essential role to recruit ubiquitinated proteins to transport to aggresomes, which ultimately induces lysosomal protein degradation. We have shown that inhibition of proteasomes with bortezomib and of aggresomes with HDAC6 inhibitor Tubacin demonstrated significant cytotoxicity in MM cell lines and MM patient tumor cells in vitro (Hideshima T et al., PNAS2005, 102: 8597–8572). In this study, we further examined the biologic significance of HDAC6 inhibition by Tubacin in MM cells. We found that HDAC6 is constitutively associated with heat shock protein (Hsp) 90 in MM cell lines which is enhanced by Tubacin, as evidenced by co-immunoprecipitation. Since Akt and STAT3 have been shown to play important role in proliferation, anti-apoptosis, and drug resistance in MM cells; and all are client proteins of Hsp90, we next further examined whether inhibition of HDAC6 could modulate activities of these proteins via Hsp90. Importantly, Tubacin enhanced phosphorylation of Akt, associated with augmentation of Hsp90 acetylation. Hsp90 inhibitor 17-AAG downregulated Akt phosphorylation associated with enhanced interaction of Hsp90 with Akt, which was partially blocked by Tubacin. On the other hand, 17-AAG did not enhance acetylation of α-tubulin or ubiquitination of proteins, suggesting that Hsp90 does not affect HDAC6 function. Furthermore, we found that STAT3 is also constitutively associated with Hsp90. Importantly, both Tubacin and 17-AAG inhibit phosphorylation of STAT3 in a dose- and time-dependent fashion in MM cells. Taken together, our data indicate that HDAC6 has an important role not only in aggresomal protein degradation, but also in MM cell pathogenesis by modulating Akt and STAT3 signaling cascades via Hsp90 acetylation in MM cells.
Azacitidine Down-Regulates Both IL-6 Signalling and NFkB Activity in Human Myeloma Cells.
Authors: Khong Tiffany , Spencer Andrew 
Published: October 2019

Abstract
We have previously demonstrated that Azacitidine (AZA) at clinically achievable concentrations (<10μM) induces apoptosis in a range of genetically heterogeneous human myeloma cell lines (HMCL) and prolongs survival in the murine 5T33 model of systemic multiple myeloma (MM). HMCL U266 demonstrates autocrine IL-6 production, constitutive Stat3 phosphorylation and is resistant to conventional chemotherapy and glucocorticoids. Using U266 we have evaluated further the effect of AZA on human MM cells. AZA treatment (5mM daily) of U266 induced apoptosis at 24, 48 and 72 hours of 28%, 43% and 52%, respectively. Immunoblot analyses showed rapid falls in IL-6R and p-Stat3 (both within 4 hours) prior to evidence of proteolysis (PARP cleavage). ELISA of AZA-treated U266 conditioned media demonstrated a reduction in IL-6 and an immediate and sustained reduction in soluble IL-6R levels, consistent with reduced IL-6 synthesis and IL-6 receptor shedding, respectively. RT-PCR confirmed that the rapid falls in IL-6 and IL6-R were not secondary to reduced transcription with relative increases of 1.7 and 2.4-fold, respectively, by 8 hours post-AZA. Flow cytometry showed an initial rise and subsequent modest fall in both membrane and cytosolic gp130 while RT-PCR and immunoblot analyses showed a rapid fall in both SOCS3 transcriptional activity and protein levels, respectively. Time-course experiments with AZA 5μM and the addition of either physiological (100pg/ml) or supra-physiological (3ng/ml) concentrations of exogenous IL-6 over a 96-hour period demonstrated that 3ng/ml but not 100pg/ml IL-6 was capable of partially restoring Stat3 phosphorylation but that neither concentration of exogenous IL-6 prevented AZA-induced cell death. NFkB activity was evaluated with a luciferase reporter enzyme system following treatment for 24 hours with AZA 1μM, 5μM and 10μM. Marked suppression of NFkB activity was seen at both the 5μM and 10μM dose levels. Flow cytometric evaluation of phospho-IkB and IkB showed a modest decline in the pIkB:IkB ratio over the same period. Finally, combined treatment of U266 with AZA and dexamethasone demonstrated synergistic killing (synergism quotient = 1.2) when AZA was administered prior to dexamethasone but not in the reverse order. We conclude that AZA induces down-regulation of both IL-6 signalling and NFkB activity. Furthermore, our data suggest that the former effect is most likely mediated by rapid AZA-induced inhibition of protein synthesis. The mechanism of NFkB inhibition requires further clarification. AZA represents a potentially novel therapeutic approach to MM.
Stromal Cell-Derived Factor-1 Stimulates the Growth of Myeloma Cells in Concert with IL-6 in Both Autocrine and Paracrine Manners.
Authors: Kim Seong-Woo , Lee Hyo-Jin , Yun Hwan-Jung , Kim Samyong , Jo Deog-Yeon 
Published: October 2019

Abstract
Stromal cell-derived factor-1 (SDF-1), which is constitutively expressed and produced in bone marrow (BM) stromal cells (BMSCs), functions via its receptor CXCR4 in the homing of myeloma cells to the bone marrow. In addition, SDF-1 inhibits dexamethasone-induced cell apoptosis. However, it has not been clarified whether SDF-1 stimulates myeloma cell growth. In this study, we explored the possibility that SDF-1 functions as an autocrine growth factor of myeloma cells. RPMI-8226 myeloma cells and primary CD138+ cells obtained from the BM of patients with multiple myeloma expressed SDF-1 mRNA, as revealed by RT-PCR analysis. SDF-1 was detected by ELISA in the supernatants of cultures of these cells grown for three days in serum-free medium, which indicates that myeloma cells are able to produce and secrete SDF-1. SDF-1 (added at a concentration of up to 100 ng/mL) alone had no discernible effects on the proliferation of RPMI-8226 cells and primary BM CD138+ cells in a serum-free condition. Interleukin-6 (IL-6) stimulated a moderate level of proliferation of these cells, while the combination of SDF-1 and IL-6 markedly enhanced the proliferation of these cells. IL-6 did not influence CXCR4 expression on myeloma cells. However, SDF-1 up-regulated significantly the expression of IL-6 receptor in these cells, as determined by Western blot analysis. SDF-1 and IL-6 enhanced in an additive fashion the phosphorylation of Akt and ERK1/2, but did not increase the phosphorylation of STAT3. Pertussis toxin (PTX, 200 ng/mL) and AMD3100 (100 nM), which is a specific antagonist of CXCR4, inhibited the proliferation of RPMI-8226 cells by 35% and 37%, respectively, in cultures grown for four days under serum-free conditions. To determine the effects of blocking the SDF-1/CXCR4 axis on the growth of myeloma cells in the presence of BMSCs, RPMI-8226 cells were co-cultured with MS-5 murine BMSCs. The addition of PTX and AMD3100 (100 nM) twenty-four hours after the beginning of the co-culture markedly decreased the number and size of cobblestone-like areas underneath the stromal cells. These results indicate that myeloma cells can produce SDF-1, and that SDF-1 in concert with IL-6 enhances the proliferation of myeloma cells in both a paracrine and autocrine manner.