bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of Fluorescence In Situ Hybridization for EGR1 Vs CSF1R for the Detection of Del(5q) in Myelodysplasia and Acute Myeloid Leukemia.
Authors: Sun Yang , Cook James R. 
Published: April 2020

Abstract
The detection of del(5q) in myelodysplastic syndrome (MDS) provides useful information to guide the choice of therapy, given the efficacy of lenalidomide in cases containing this abnormality. Fluorescence in situ hybridization (FISH) analysis offers the opportunity to specifically detect chromosomal abnormalities much more rapidly than metaphase cytogenetics, which may have a turnaround time of several weeks. However, it is currently unclear which chromosomal loci are the most appropriate to examine for detection of del(5q) in routine practice. The breakpoints on chromosome 5q are heterogeneous and two commonly deleted regions (CDR) have been described. The first CDR occurs in acute myeloid leukemia (AML) and high grade MDS and encompasses a region at 5q31 including the EGR1 locus. A second CDR, occurring in at least some cases reported as 5q- syndrome, centers around 5q33 and includes the CSF1R locus. We therefore examined whether FISH studies for EGR1, CSF1R, or a combination of both probes would provide the greatest clinical utility for detection of del(5q). 51 cases of myeloid neoplasms with del(5q) by metaphase cytogenetics were analyzed, including 5q- syndrome (n=8), refractory anemia with excess blasts (RAEB, n=8), refractory cytopenia with multilineage dysplasia (RCMD, n=6), MDS unclassifiable (n=1), therapy related MDS (n=1), myelodysplastic/myeloproliferative overlap syndromes (MDS/MPD, n=6), and AML (n=21). FISH studies using EGR1/D5S23, D5S721 and CSF1R/D5S23, D5S721 probes (Abbot Molecular, Abbot Park, IL) were performed on archival bone marrows (45 coverslip aspirate smears, 4 cytogenetic culture cell pellets, and 2 formalin fixed paraffin embedded clot sections). Normal ranges were established for each probe by analysis of appropriate negative control samples. Deletion of the EGR1 locus was detected in 49/51 (96%) cases, including each case of 5q- syndrome. The CSF1R locus, which could be analyzed in 48 cases, was deleted in 44 cases (92%). In cases with concordant results, a similar percentage of abnormal nuclei was identified with each probe. Two cases (1 MDS/MPD and 1 AML) displayed deletion of the EGR1 locus but a normal pattern for CSF1R. Two cases (1 AML and 1 MDS/MPD) showed no evidence of EGR1 or CSF1R deletion despite a del(5q) identified by metaphase cytogenetics. In conclusion, FISH for EGR1 is sufficient to successfully detect del(5q) in the vast majority of cases of MDS and AML containing this abnormality, including at least most cases of 5q- syndrome. Additional FISH studies for the CSF1R locus did not increase the diagnostic yield. Further studies will be required to determine if deletions of 5q involving EGR1 but not CSF1R influence the response to lenalidomide. A small number of cases of del(5q) are detected only by metaphase cytogenetics, possibly due to a small number of abnormal cells present prior to in vitro culture.
Characteristics and Outcome of MDS with Isolated Del 20q: The GFM Experience in 64 Cases.
Authors: Braun Thorsten , de Botton Stephane , Taksin Anne-Laure , Park Sophie , Beyne-Rauzy Odile , Chevret Sylvie , Fontenay Michaela , Cassinat Bruno , Lazareth Anne , Eclache Virginie , Terre Chrisitine , Vey Norbert , Guerci Agnes , Dreyfus Francois , Bordessoule Dominique , Stamatoullas Aspasia , Castaigne Sylvie , Fenaux Pierre , Ades Lionel 
Published: April 2020

Abstract
Background: Isolated del 20q is common in MDS, and considered of good prognosis in the IPSS, but no large series of MDS with del 20q have been reported.
Methods: We studied characteristics and outcome of 64 MDS patients (pts) with isolated del 20q diagnosed between 1990 and 2008 in 5 centers of the GFM. They were also compared with MDS without del 20q included in the GFM registry (containing 995 karyotyped MDS diagnosed since 2003).
Results: Median age of the 64 MDS with isolated del 20q was 72 (48–92), with 41 males and 23 females. They included 34 RA, 8 RARS, 16 RAEB, 4 CMML and 2 RAEB-T. Fifty two percent of them had less than 5% of marrow blasts, compared to 34% in the non del 20q pts included in the GFM registry (p=0.006). Despite a significantly higher proportion of low and int-1 IPSS risk (90% vs 78%, p=0.021) the isolated del 20q cohort had lower platelet count (mean 156 vs 212 G/l, p=0.014) and higher reticulocyte count (mean 84 vs 52 G/l, p=0.012) than non del 20 q pts included in the GFM. No significant difference was found for age, gender, FAB, ANC and Hb level between isolated del 20q and non del 20q pts. When the analysis was restricted to pts with less than 5% marrow blasts, the same differences were seen between isolated del 20q and non del 20q pts. On the other hand, 12 (19%) of the pts with isolated del 20q had Hb &gt; 12g/dl and platelets &lt; 100 G/L vs 80 of non del 20q patients (p=0.007). The 12 pts with isolated del 20q and plts &lt;100G/L without anemia, were mostly men (9/12), with marrow blasts &lt;5% in 83% (10/12), and all (100%) had ANC &gt;1 G/l. None of those del 20q pts progressed to AML (compared to 17% of isolated del 20 q pts with Hb &lt;12 g/dl and/or plts &gt;100 G/l) and their median overall survival was 39 months vs 29 months, respectively (p=0.32). Median survival of MDS with isolated del 20q was 37 months and was significantly longer in pts with marrow blasts &lt;5% (12 vs 60 months, p=0,0001), with low and int-1 IPSS score (39 vs 11 months, p=0.01) and with Hb &gt;11 g/dl (53 vs 16 months, p=0.002). Survival could not be compared between isolated del 20q and non del 20q pts because of the short observation period of the GFM registry.
Conclusion: MDS with isolated del 20q are characterized by usual absence of excess of blasts lower platelets and less anemia than other MDS. About 20% of them present with isolated thrombocytopenia. Prognosis is favorable only in the absence of excess of blasts.
Large Scale Evaluation of Genetic Variation and the Risk of Multiple Myeloma.
Authors: Berndt Sonja I , Johnson David C , Crowley John , Durie Brian G , Hoover Robert , Katz Michael , Rothman Nathaniel , Van Ness Brian G. , Baris Dalsu , Morgan Gareth J 
Published: April 2020

Abstract
Genetic factors are thought to influence susceptibility to multiple myeloma, but most published studies to date have been small and limited in scope. To identify genetic polymorphisms associated with myeloma risk, we conducted a case-control study of 976 Caucasian myeloma cases enrolled from clinical trials as part of the International Myeloma Foundation’s Bank On A Cure® initiative and 3692 Caucasian controls from the three cohorts [Nurses’ Health Study (NHS), Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO), and 1958 British Birth Cohort (BC58)] with genome-wide scan data. A candidate gene approach was taken with a preference given to single nucleotide polymorphisms (SNPs) in coding or regulatory regions. A total of 1097 SNPs with a minor allele frequency ≥1% were genotyped in the cases and at least one control population. In order to increase our statistical power, SNPs not genotyped in NHS and PLCO were imputed from the genome scan with MACH using the HapMap CEU population as a referent and included in the analysis if the quality control r2 was high (r2 ≥0.9). Logistic regression was used to estimate the odds ratios (ORs) and 95% confidence intervals (95% CIs) adjusting for age, sex, and country as appropriate. We found 26 loci to be associated with myeloma risk with P &lt; 0.01. Of particular interest, we observed an increased risk of myeloma with variants in two genes involved in the metabolism of pyrimidines, DPYD and MTHFR. An increased risk of myeloma was found with two independent SNPs, rs1023244 and rs1399291, in DPYD (ORperGallele = 1.43, 95% CI: 1.16–1.76, P = 0.0008 and ORperTallele = 1.18, 95% CI: 1.06–1.31, P = 0.003, respectively) and with the MTHFR high activity 677C allele (rs1801133, ORperC allele = 1.18, 95% CI: 1.05–1.33, P = 0.006). We also observed significant associations for nonsynonymous SNPs in genes involved in cell cycle checkpoint regulation (ATR, P = 0.009; ZAK, P = 0.007) and the DNA damage bypass pathway (REV3L, P = 0.008), suggesting that alterations in DNA damage mediation may modulate myeloma susceptibility. In conclusion, this large study found SNPs in several pathways, including pyrimidine metabolism and DNA damage mediation, to be associated with myeloma risk. Additional studies are needed to replicate these findings and to further explore genetic variation in these regions.
Wnt3 Signaling Augments integrinβ1-Dependent Adhesion-Mediated Drug Resistance of Multiple Myeloma in Autocrine and Paracrine Manner.
Authors: Kobune Masayoshi , Murase Kazuyuki , Iyama Satoshi , Sato Tsutomu , Kawano Yutaka , Kikuchi Shohei , Miyanishi Koji , Takimoto Rishu , Kato Junji 
Published: April 2020

Abstract
Adhesion of myeloma cells to BM stromal cells is now considered to play a critical role in chemo-resistance. However, little is known about the molecular mechanism of cell adhesion mediated drug resistance (CAM-DR) in MM. In this study, we focused on relationship between drug resistance and expression of Wnts, the factor regulating the cell adhesion and proliferation, in myeloma cells. To gain insight into involvement of Wnt signaling in CAM-DR, we first screened the expression of Wnt family in myeloma cell lines (RPMI8226, ARH77, KMS-5, and MM1S) by reverse transcription-polymerase chain reaction analysis. Although the mRNAs of Wnt2b, Wnt7a and Wnt10b were variably expressed in some of myeloma cell lines, Wnt3 mRNA was detected in all the myeloma cells examined. KMS-5 and ARH77, which highly expressed Wnt3 protein, tightly adhered to human BM stromal cells and accumulation of beta-catenin and GTP-bounded RhoA was observed in these myeloma cell lines. Conversely, RPMI8226 and MM1S, which modestly expressed Wnt3 protein, rather weakly adhered to human BM stromal cells. We then examined the relevance of Wnt3 expression to adhesive property to stromal cells and to CAM-DR of myeloma cells. KMS-5 and ARH-77 exhibited apparent CAM-DR against Doxorubicin. This CAM-DR was significantly reduced by anti-integrinβ1 antibody, a Wnt-receptor competitor, secreted Frizzled related protein-1 and Rho kinase inhibitor (Y27632 and OH-fasudil), but not by the specific inhibitor of canonical signaling (DKK-1), indicating that Wnt-mediated CAM-DR which is dependent on integrinβ1- mediated attachment to stromal cells is induced by Wnt/Rho pathway signal. This CAM-DR for doxorubicin was also significantly reduced by Wnt3 siRNA transfer to KMS-5. On the other hand, the cell adhesion of MM1S was dramatically augmented by addition of Wnt3 containing conditioned medium (CM) and suppressed by integrinα4 or β1 antibody (VLA4)(Fig 1). Furthermore, CAM-DR of MM1S was significantly augmented by Wnt3 CM or adhesion of mesenchymal stem cells which expressed Wnt3, but not BM stromal cells which did not express Wnt3 mRNA. These results suggest that adhesion of myeloma cells on stromal cells was regulated by Wnt signaling in autocrine or paracrine manner. The Wnt3 signaling pathway could be a promising molecular target to overcome CAM-DR.
Glucocorticoid Receptor Expression Correlates with Clinical Outcome in Myeloma Patients Treated with Glucocorticoid-Containing Regimens.
Authors: Ma Shuo , Zhan Fenghuang , Ma Chunguang , Tessel Michael , Zhang Lijun , Rajkumar S. Vincent , Singhal Seema , Barlogie Bart , Shaughnessy John , Rosen Steven T. , Krett Nancy 
Published: April 2020

Abstract
Glucocorticoid (GC) is an effective anti-myeloma agent by directly inducing apoptosis and growth inhibition in myeloma cells. The anti-myeloma action of GC is mediated by its intracellular receptor, the glucocorticoid receptor (GR). The response to GC is variable among myeloma patients and resistance to glucocorticoid treatment develops almost invariably. The mechanism regulating glucocorticoid-sensitivity in not well understood, although it has been hypothesized that the regulation might occur at both the receptor (GR) and post-receptor level. In MM.1 myeloma cell lines, we observed significant down-regulation of GR expression as the cells develop GC-resistance, suggesting that GR expression level is an important determinant of GC-response. Here we present evidence that GR expression level directly correlates with the clinical outcome in myeloma patients who were treated with GC-containing regimens. First, using a quantitative real-time PCR analysis, we performed a retrospective analysis of GR gene expression on bone marrow plasma cell samples from 35 patients with newly diagnosed myeloma who were then treated on the ECOG study E1A00. This is a phase 3 trial which randomized patients to dexamethasone (dex) alone vs. dex in combination with thalidomide. Pre-treatment expression levels of GR-alpha, GR-beta and GR-P splicing isoforms were measured for 35 patients, of whom, 10 had post-treatment samples available. Interestingly, a large variation of the GR expression levels was observed across the patient samples, with the GR-alpha being the predominant isoform. However, a clear correlation between GR expression and treatment response was not seen, likely due to the small sample size. To access a larger patient pool, we retrospectively analyzed the GR expression levels using the cDNA microarray data from the Arkansas group on 351 newly diagnosed myeloma patients treated with Total Therapy 2, of which dexamethasone was an essential component throughout the treatment course. Of the 351 patients, both event-free (EFS) and overall survivals (OS) were compared between 49 patients (14%) with low levels of GR expression and the rest (86%) with high levels of GR expression. Strikingly, the cases with low levels of GR expression were associated with statistically significantly worse EFS and OS compared to those with high levels of GR expression, with the 5-year EFS of 20% vs. 52% and OS of 34% vs. 68%, P &lt; 0.01. In summary, we have shown for the first time that the baseline level of glucocorticoid receptor expression in newly diagnosed myeloma patients predicts their clinical response to GC-containing regimen and the overall clinical outcome.
The CXCR4 Antagonist AMD3100 Stimulates the Proliferation of Myeloma Cells.
Authors: Kim Ha-Yon , Kim Seong-Woo , Lee Hyo-Jin , Yun Hwan-Jung , Kim Samyong , Jo Deog-Yeon 
Published: April 2020

Abstract
AMD3100, an antagonist of the chemokine receptor CXCR4, is about to be used clinically for the peripheral mobilization of hematopoietic stem cells, especially in patients with lymphoma and multiple myeloma. However, AMD3100 has been shown to activate a G protein coupled with CXCR4 and thus acts as a partial CXCR4 agonist in vitro. Although stromal cell-derived factor-1 (SDF-1) alone has minimal or negligible effects on the growth and survival of myeloma cells in vitro, many reports are consistent with the SDF-1/CXCR4 axis being involved in the progression of myeloma. Therefore, it is necessary to address the question of whether AMD3100 functions as a partial agonist for CXCR4 in myeloma cells, before it is released for wide clinical application. In this study, we explored whether AMD3100 affects the proliferation and survival of myeloma cells in vitro. As demonstrated previously, AMD3100 markedly inhibited the SDF-1 induced chemotaxis of myeloma cells, including three cell lines (RPMI8226, U266, and ARH77 cells), and CD138+ primary human myeloma cells. AMD3100 also induced internalization of CXCR4. SDF-1 alone did not stimulate the proliferation of these myeloma cells, nor did it rescue the cells from apoptosis induced by serum deprivation. By contrast, AMD3100 at 10−5M enhanced the proliferation of all three myeloma cell lines in serum-free condition by up to 2-folds in 3-day cultures, which was abrogated by pretreating the cells with pertussis toxin (PTX). This phenomenon was also observed with CD138+ primary human myeloma cells. In addition, AMD3100 enhanced the proliferation of U266 and ARH77 cells induced by interleukin-6 (IL-6). AMD3100 partially inhibited the apoptosis induced by serum deprivation, and the anti-apoptotic effect of AMD3100 was further enhanced in the presence of IL-6. AMD3100 on its own induced the phosphorylation of Akt and ERK1/2 but not Stat3 and p38/MAPK in RPMI8226 and U266 cells. The phosphorylation was also inhibited by pretreating the cells with PTX. The signal blocking agents wortmannin, PD98056, SB203580, and rapamycin did not affect the AMD3100-induced proliferation of RPMI8226 cells. LY294002 at concentrations not inhibiting spontaneous proliferation (5 μM or less) did not affect AMD3100-induced proliferation of the cells. AG490 inhibited spontaneous proliferation and AMD3100-induced proliferation of the cells in a dose-dependent manner. Accordingly, the signal-blocking effects were unclear. However, these results show that AMD3100 partially overcame the strong growth inhibition of myeloma cells induced by AG490. Our results indicate that AMD3100 can induce the phosphorylation of signaling molecules and stimulate the proliferation of myeloma cells, through signaling via G-protein pathway.
Proteomic Profiling of Multiple Myeloma: Correlation of Protein and Gene Expression Data.
Authors: Edmondson Ricky D , Thyparambil Sheeno P , MacLeod Veronica , Barlogie Bart , Shaughnessy John D. 
Published: April 2020

Abstract
Although melphalan-based autologous stem cell transplantation has improved prognosis for patients diagnosed with Multiple Myeloma, survival varies from a few months to more than 15 years with an individual’s risk not accurately predicted with standard prognostic variables. Correlating genome-wide mRNA expression profiles in purified myeloma cells with outcome, we recently showed that that the differential expression of 70 genes could identify patients at high risk for early disease related death [1]. The utility of a high throughput proteomics platform in the analysis of clinical samples has great potential but as of yet none have been firmly established. Herein, we describe the use of such a platform and its utility in stratifying patients with Multiple Myeloma in terms of high and low risk disease. Preliminary analysis indicates that the proteomics data can separate the patients into risk groups, although the proteins responsible for the assignment are not identical to the 70 genes identified in the gene expression profiling experiments. In addition to the proteomic analysis of plasma cells enriched using anti-CD138 immunomagnetic beads from mononuclear cell fractions of bone marrow aspirates from newly diagnosed myeloma patients; we have performed (in triplicate) LCMS profiling on plasma cells from 30 patients isolated prior to and 48 hours after a single test-dose application of bortezomib at 1.0mg/m2. An aliquot of 100,000 plasma cells was enzymatically digested with trypsin and a fraction (~5,000 cells) analyzed using our proteomics platform (an Eksigent nanoHPLC coupled to a ThermoElectron LTQ-Orbitrap with data analyzed using the Elucidator software package from Rosetta Biosoftware). The correlation of the proteomic profiles to gene expression profiles and clinical parameters will be presented. The analysis of proteins that were observed to change (p&lt;0.01) in abundance after the single agent dose of the proteasome inhibitor bortezomib yielded an unanticipated finding; the abundance of 30 proteins associated with the proteasome were observed to increase in a subset of patients. The majority of the patients with the increased levels of proteasome related proteins are predicted by GEP to have high risk disease. The proteomic data will be discussed in terms of its utility in the identification of activated pathways as well as in the development of a prognostic indicator as was achieved using gene expression profiling.
Agonist Anti-TRAILR1 Human Mab, Hgs-ETR1, Induces Myeloma Cell Death Mainly through the Intrinsic Pathway of Apoptosis.
Authors: Ménoret Emmanuelle , Surget Sylvanie , Gomez-Bougie Patricia , Bataille Régis , Amiot Martine , Pellat-Deceunynck Catherine 
Published: April 2020

Abstract
We previously reported that agonist anti-TRAILR mAbs (HGS-ETR1) induced cell death of myeloma cells through both the extrinsic and intrinsic pathway. Although caspase 8 activation was fully required to initiate death signaling, it was unclear whether intrinsic pathway played or not a major role in apoptosis execution. In the current work, we evaluated the contribution of the intrinsic pathway in a large panel of HMCL (n=10). To address this question, we monitored mitochondrial depolarization and the contribution of caspase 9. Apoptosis and mitochondrial polarization were both evaluated by flow cytometry. Kinetic analysis of mitochondrial polarization (JC-1) shows that all mitochondria were fully depolarized after only 4h following HGS-ETR1 mAb addition: the depolarization started early, as soon as 1h. In good agreement with mitochondrial depolarization, we observed that specific inhibitor of caspase 9 activity (z-LEHD-fmk) always prevented at least 50% of death (and up to 90%). Our data show that in all cell lines evaluated, activation of the intrinsic pathway was required for cell death even with high concentrations of mAb. We previously showed that anti-TRAILR apoptosis was always associated with an early and massive Mcl-1 cleavage (Menoret et al Blood2006, 108:1346). In myeloma cells, Mcl-1 is physically associated with Bim and thus prevents Bim to induce mitochondrial cell death (Gomez-Bougie EJI 2004, 34;3156). By immunoprecipitation, we show that, in the presence of HGS-ETR1 mAb, Mcl-1/Bim complexes were disrupted and Bim was free to activate Bax. Bim is also found to be complexed to both Bcl-XL and Bcl-2 in viable cells. During anti-TRAILR mAb induced apoptosis, Bcl-2 and Bcl-XL were also cleaved and full length Bcl-XL, as Mcl-1, fully disappeared indicating that BcL-XL decrease probably participated to the increase of free Bim. Furthermore, immunoprecipitation of Bax showed that Bax was associated with Bcl-2 (and not with Mcl-1 or Bcl-XL) and this association was reduced in apoptotic conditions. These data show that apoptosis induced by anti-TRAILR mAb mainly involved the intrinsic pathway through disruption of both anti-apoptotic/Bim and Bcl-2/bax complexes. This result suggests that the association of anti-TRAILR mAb with melphalan or bortezomib would not synergistically increase anti-TRAILR mAb efficiency and that anti-TRAILR mAb should be rather combined with drugs acting independently of the intrinsic pathway.