bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Three Novel Chromosomal Translocation Partners Involving the Immunoglobulin Loci in Newly Diagnosed Myeloma and Human Myeloma Cell Lines.
Authors: Hanamura Ichiro , Iida Shinsuke , Ueda Ryuzo , Kuehl Michael , Cullraro Contance , Bergsagel Leif , Sawyer Jeffrey , Barlogie Bart , Shaughnessy John 
Published: April 2020

Abstract
Chromosomal translocations involving the immunoglobulin heavy-chain (IgH) locus at 14q32 are genetic hallmarks of human multiple myeloma (MM) and are likely primary oncogenic events in myeloma transformation. Common recurrent translocation partner genes of IgH account for 40–50% of cases and these rearrangements result in the transcriptional dysregulation of various proto-oncogenes such as CCND1, CCND3, FGFR3/MMSET, c-MAF, MAFB, MUM 1(IRF-4), c-MYC and it is becoming clear that these dysregulated genes play important roles in the pathogenesis and the treatment outcome of MM. Although the frequency of other partner genes of IgH translocations may be rare (&lt;1%), it is possible that many genes not currently identified as translocation partners are functionally converged into several signal pathways and play important roles as well. Through global gene expression profiling (GEP) studies of a large population of newly diagnosed MM, we have recently reported that dysregulation of any one of the three D-type cyclins is seen in almost all newly diagnosed MM and most patients with high levels of CCND2 expression was highly correlated with expression of c-MAF, MAFB or FGFR3/MMSET. In order to identify novel translocation partner genes in MM we combined GEP and fluorescence in situ hybridization (FISH) analysis in a large population of newly diagnsoed myeloma. We investigated whether IGH-CCND2 and IGH-MAFA translocations are seen in patients exhibiting high levels of CCND2 expression but not expressing c-MAF, MAFB, or FGFR3/MMSET. We also looked for a breakpoint and targeted gene at a non-random breakpoint at the 11q23 locus in 20 MM patients with 11q23 abnormalities detected by conventional G-banding. Here we report evidence of an IGH-CCND2 fusion in a newly diagnosed case of MM and an IGH-MAFA fusion in a newly diagnosed MM and an MGUS and also mapped the 11q23 breakpoint to within 100 kb in a newly diagnosed MM and a human myeloma cell line, NCU-MM-1 and identified an IGL Pou2AF1 fusion in the NCU-MM-1 line. Importantly these translcoatios do not involve the MLL or ATM genes. These three genes were all highly expressed as detected by GEP or quantitive RT-PCR in the samples in which these genes were translocated to IG loci. Although the frequency of these 3 translocations are very rare, they further the concept of a critical role of CCND2 expression in myelomegenesis and identifed the transcriptional coactivator Pou2AF1, which specifically associates with either OCT1 or OCT2, in the B-cell response to antigens and required for the formation of germinal centers, as a potential new translocation target in myeloma. Finally, our strategy of combining GEP signatures and cytogenetic analyses appears to be a practical means of identifying new IG- mediated translocations in myeloma.
Frequent Gains of Chromosome Arm 1q21 in Plasma Cell Dyscrasia Detected by Fluorescence In Situ Hybridization: Incidence Increases from MGUS to Relapsed Myeloma and Is Related to Disease Progression and Prognosis.
Authors: Hanamura Ichiro , Stewart Peter , Cavallo Frederica , Burington Bart , Rasmussen Erik , Crowley John , Zhan Fenghuang , Huang Yongsheng , Barlogie Bart , Shaughnessy John 
Published: April 2020

Abstract
Background: High-dose therapy with autologous stem cell transplantation (HDT-ASCT), effecting complete remissions in &gt; 40% with median overall survival exceeding 6 years, has become the standard of care for patients with multiple myeloma (MM). Poor outcome following HDT-ASCT has been linked most strongly to metaphase cytogenetic abnormalities (CA). Tandem duplications and jumping translocations of the chromosome 1q21 region is a recurrent event in advanced MM and 1q21 amplification (amp1q21) and high expression of the CKS1B gene (mapping to 1q21), both signify poor survival in MM treated with HDT-ASCT. Smoldering multiple myeloma (SMM) and monoclonal gammopathy of undetermined significance (MGUS) have many of the genetic hallmarks of overt MM, but especially MGUS rarely proceeds to MM requiring treatment. The aim of this study was determine if amp1q21 relates to disease progression and prognosis.
Patients and Methods: 467 untreated patients with MGUS (n = 14), SMM (n = 32) and MM (n = 421) as well as 42 with relapsed MM were examined by means of triple color interphase fluorescence in situ hybridization (TRI-FISH) of the CKS1B locus.
Results: The frequency of ampq21 was 0% in 14 patients with MGUS, 43% in 32 with SMM, 43% in 421 with newly diagnosed MM and 78% in 42 with relapsed MM (p&lt;0.001; untreated vs relapsed MM). Among SMM patients, 11 of 14 (78%) with amp1q21 but only 2 of 18 (11%) without amp1q21 progressed to active MM (p&lt;0.001) (median follow-up, 44 mo; range, 0–120 mo). In 421 MM patients treated on Total Therapy 2 (high-dose melphalan-based tandem autotransplants +/− thalidomide), 5-yr overall survival/event-free survival were 75%/61% in without amp1q21 compared to 48%/34% with amp1q21 (both p&lt;0.001). Among 42 relapses, amp1q21 was present in 20/21 occurring on thalidomide vs 13/21 on no thalidomide (p=0.020); the median post-relapse survival in those with amp1q21 was only 11 mo vs 25 mo in patients without amp1q21 (p=0.02).
Conclusion: (1) Amp1q21 was absent in MGUS and present in &gt;40% of symptomatic MM and SMM. (2) in SMM, amp1q21 was associated with a higher risk of conversion to MM. (3) In overt MM, amp1q21 conferred inferior event-free and overall survival.
Gains on Chromosome Arm 9q Represent a Novel and Independent Marker of Adverse Prognosis in Multiple Myeloma Patients Receiving Upfront High-Dose Chemotherapy and Autologous Stem Cell Transplantation.
Authors: Liebisch Peter , Benner Axel , Tschajka Kathrin , Wendl Christiane , Goldschmidt Hartmut , Döhner Hartmut 
Published: April 2020

Abstract
Introduction: Genomic aberrations represent important prognostic markers in many hematological cancers. In multiple myeloma (MM), chromosome 13q and 17p deletions (13q−, 17p−) have emerged as important outcome predictors that indicate a dismal prognosis. Other chromosomal abnormalities have been discussed as prognostic markers in this disease but came not out as independent variables when they were tested in a multivariable fashion. However, the complexity of genomic rearrangements and the clinical heterogeneity seen in malignant plasma cell disorders argue against 13q− and 17p− as the sole genomic change of prognostic relevance. The significance of chromosome arm 1q, 9q, and 11q extra copies - three frequent genomic imbalances in MM - is undetermined.
Material and Methods: 90 patients (pts.) treated with one or two cycles of high-dose chemotherapy (HD-CTX) followed by autologous stem cell transplantation (ASCT) at a single center were analyzed by tri-color FISH and five DNA probes mapping to chromosome bands 1q21, 9q34, 11q25, 13q14, and 17p13. A multivariable analysis (Cox proportional hazards regression model) including genetic and clinical variables was performed.
Results: The most frequent aberrations in the present series were (in order of decreasing prevalence): +1q (n=39/78, 50.0%), +9q (n=38/78, 48.7%), 13q− (n=42/90, 46.6%), and +11q (n=39/85, 45.9%). 17p− was identified in only 3 out of 88 patients (3.4%), while +17p were present in 13 out of 88 patients (14.8%). The median follow-up time was 37 months (m) and the median event free survival (EFS) and overall survival (OS) time from first ASCT of the entire cohort was 26 m and 71 m, respectively. Multivariable analysis including genetic aberrations, ß2-microblobulin, albumin, LDH, Salmon/Durie stage, and age at time of diagnosis revealed +9q and 13q− as the only independent predictors of EFS (p=0.003 and p=0.01, respectively). The mEFS in patients with 13q− was 19.0 m and 20.7 m in patients with +9q. In patients with concurrent +9q and 13q−, mEFS was only 12.2 m. In patients lacking these two abnormalities, mEFS was not reached. OS was not significantly influenced by any genetic or clinical variable in our series, most likely due to effective salvage treatment after relapse.
Conclusions: +9q represents a novel and independent marker of adverse prognosis in MM. A single FISH experiment applying two DNA probes allows easy and rapid assessment of outcome in patients with malignant plasma cell disorders.
Studies of the Bcl Inhibitor GX15-070 in Multiple Myeloma Demonstrate Substantial Pre-Clinical Activity.
Authors: Stewart A. Keith , Li Zhi Hua , Wen Xiao Yan , Rauw Jennifer , Pathangey Latha , Bergsagel P. Leif , Trudel Suzanne 
Published: April 2020

Abstract
GX15-070 is a small-molecule antagonist of the BH3-binding groove of the Bcl-2 family of proteins and can thus induce apoptosis by inhibiting the interaction between pro- and anti-apoptotic proteins. In addition, GX15-070 activates a cellular pathway, which leads to the activation of the pro-apoptotic BH3-only protein Bim, a natural cellular inhibitor of anti-apoptotic members. Together, the direct inhibition of anti-apoptotic Bcl-2 family proteins and the activation of pro-apoptotic Bim by GX15-070 can sensitize tumor cells to apoptosis. Bcl-2 family members, particularly Bcl-XL and Mcl-1 have been implicated in the survival of myeloma cells and may be responsible, in part, for drug resistance. Consequently, GX15-070 was tested against the MMRC 12 validation cell line panel, a panel of 12 standardized and annotated human myeloma cell lines (HMCLs). Western blots preformed on 8 HMCLs confirmed expression of Bcl-2, Mcl-1 and/or Bcl-XL. Basal levels of Bcl-2 and MCl-1 were relatively similar across all HMCLs studied whereas levels of Bcl-XL were variable GX15-070 inhibited the viability of all 12 myeloma cell lines as determined by MTT assay. The mean IC50 was 215 nM; range (6.4-678nM) and sensitivity did not correlate with level of Bcl-2, MCL-1 or Bcl-XL expression. On the other hand cytotoxicity to normal human blood lymphocytes, stroma and bone marrow CFUs was not observed at concentrations which were effectively cytotoxic to HMCL. Time course experiments demonstrate that apoptosis and cleavage of caspase 3 began by 12 hours and continued to increase over a 96 hour period. Co-culture with human bone marrow stroma cells (BMSCs) failed to protect HMCL from GX15-070 induced cytotoxicity. Similarly, potent MM growth factors, Interleukin 6 (IL-6) and Insulin like growth factor (IGF)-1 did not confer resistance to GX15-070. Of interest, overexpression of Bcl-2 has been shown to attenuate PS-341-induced cell death (Mitisades); however, the combination of PS-341 and GX15-070 did not reveal synergistic interaction in preliminary results. Studies looking at other drug combinations are planned. Finally, GX15-070 produced cytotoxic responses in the first primary myeloma sample tested with further testing ongoing. Mouse xenograft studies are also ongoing and will be presented. These studies indicate extensive pre-clinical activity of GX15-070 in multiple myeloma and set the stage for planned Phase II clinical trials.
BCL6 Is Regulated by p53 through a Response Element Frequently Disrupted in B-Cell Non-Hodgkin’s Lymphoma.
Authors: Margalit Ofer , Amram Hila , Amariglio Ninette , Simon Amos J. , Granot Galit , Minsky Neri , Shaklai Sigal , Shimoni Avichai , Harmelin Alon , Givol David , Shohat Mordechai , Oren Moshe , Rechavi Gideon 
Published: April 2020

Abstract
The BCL6 transcriptional repressor mediates survival, proliferation and differentiation blockade of B-cells during the germinal-center reaction, and is frequently misregulated in B-cell non-Hodgkin’s lymphoma (BNHL). The p53 tumor-suppressor gene plays a crucial role in tumorigenesis. In a previous study done in our laboratory, DNA microarrays were used to identify primary p53 target genes, one of which was BCL6 (Kannan et al. 2001. Oncogene20: 2225). The BCL6 intron 1 contains a region in which 3 types of genetic alterations are frequent in BNHL: chromosomal translocations, point mutations and internal deletions. We therefore defined it as TMDR - Translocations, Mutations and Deletions Region. The BCL6 gene contains a p53 response element (p53RE) residing within the TMDR. This p53RE contains a motif which is known to be preferentially targeted by somatic hypermutation. This p53RE is evolutionary conserved in primates, but not in rodents. p53 binds to this RE in vitro and in vivo. Reporter assays revealed that the BCL6 intron 1 can confer p53-dependent transcriptional activation, through the p53RE. BCL6 mRNA and protein levels increased after chemo/radiotherapy in human, but not in murine tissues. The increase in BCL6 mRNA levels was attenuated by the p53 inhibitor PFT-α. Thus, we define the BCL6 gene as a new p53 target. In conjunction with the recent finding that BCL6 represses p53 gene expression (Phan & Dalla-Favera 2004. Nature 432: 635), our data define a novel autoregulatory loop, in which p53 is activated in response to genomic alterations that occur normally during B-cell maturation. In the physiological context, this p53 activation induces BCL6, which down-regulates p53 and spares the cell from p53-regulated apoptosis. This down-regulation of p53 is predicted, in turn, to attenuate BCL6, allowing it to return to baseline levels and thereby defining a tightly controlled time window for completion of B-cell maturation. Disruption of this loop may lead to deregulated BCL6 expression and promote lymphomagenesis.
Up-Regulation of c-Jun contributes to the Induction of Apoptosis by Adaphostin in Human Multiple Myeloma Cells.
Authors: Podar Klaus , Tonon Giovanni , Raab Marc S. , Sattler Martin , Abtahi Dean , Tai Yu-Tzu , LeGouill Steven , Ishitsuka Kenji , Yasui Hiroshi , Depinho Ron A. , Hideshima Teru , Chauhan Dharminder , Anderson Kenneth C. 
Published: April 2020

Abstract
Adaphostin (NCI, Bethesda, MD) has been examined as an alternative to the 2-phenylaminopyrimidine derivate imatinib mesylate, with remarkable efficacy in the treatment of chronic myeloic leukemia (CML) including (1) Bcr/Abl+ cells; (2) imatinib mesylate resistant cells; and (3) Bcr/ Abl - cells. However exact functional mechanisms by which adaphostin induces cell apoptosis are unknown. Previously we have compared effects of adaphostin, a small molecule of the tyrophostin family versus imatinib mesylate in human MM cells: significant concentration - dependent antiproliferative the effects were observed after treatment with adaphostin, but not with imatinib mesylate, on MM cells alone and adherent to BMSCs. Here, we used gene expression microarray to identify signaling molecules modulated by adaphostin. As confirmed by protein expression, several proteins, most prominently c-Jun, were upregulated in a dose- and time- dependent manner. In contrast, c-Jun upregulation was not observed in MM cells treated with imatinib mesylate. Although PARP cleavage was induced, c-Jun upregulation was also not observed after treatment with dexamethasone (0.5 and 1mM), bortezomib (5 and 20nM) or melphalan (5 and 20mM). The functional relevance of adaphostin- induced c-Jun- upregulation was next investigated using small- interfering RNAs targeting c-Jun mRNA. Adaphostin- induced caspase- cleavage is abrogated in MM cells transfected with cJun- specific RNAi, but not with mock-RNAi. Consequently, a decrease of thymidine incorporation induced by adaphostin was only observed in MM cells transfected with mockRNAi, but not c-Jun- specific RNAi. Moreover, our results also demonstrate that adaphostin- induced caspase cleavage is dependent on the expression of c-Jun, but not visa versa. These studies for the first time provide evidence for a specific adaphostin- induced c-Jun upregulation and related induction of apoptosis in MM cells. Based on these findings, ongoing studies are investigating the role of c-Jun in MM pathogenesis.
JS-K, a GST-Activated Nitric Oxide Generator, Induces Apoptosis and Overcomes In Vitro Drug Resistance in Multiple Myeloma Cells.
Authors: Kiziltepe Tanyel , Ishitsuka Kenji , Hideshima Teru , Raje Noopur , Shiraishi Norihiko , Yasui Hiroshi , Catley Laurence , Chauhan Dharminder , Saavedra Joseph E. , Keefer Larry K. , Shami Paul J. , Anderson Kenneth C. 
Published: April 2020

Abstract
Multiple myeloma (MM) is currently an incurable hematological malignancy. A major reason for the failure of currently existing therapies is the chemotherapeutic resistance acquired by the MM cells upon treatment. Overexpression of glutathione S-transferases (GST) has been shown as one possible mechanism of anti-cancer drug resistance in a broad spectrum of tumor cells. JS-K (O2-(2,4-Dinitrophenyl) 1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate) belongs to a class of pro-drugs which are designed to release nitric oxide (NO) on reaction with GST. JS-K can possibly turn GST overexpression to the tumor’s disadvantage by (1) consuming intracellular GSH and preventing drug inactivation; and (2) by exposing tumor cells to high intracellular concentrations of NO. JS-K has potent in vitro and in vivo anti-leukemic activity. The purpose of the present study is to examine the biological effects of JS-K on human MM cells. We demonstrate that JS-K has significant in vitro cytotoxicity on MM cell lines, with an IC50 of 0.3-2 mM at 48 hours. JS-K also induces cytotoxicity on cell lines that are resistant to conventional chemotherapy (i.e., MM1R, RPMI-Dox40, RPMI-LR5, RPMI-MR20). Importantly, no cytotoxic effects of JS-K were detected on peripheral blood mononuclear cells (PBMNC) obtained from healthy volunteers at these doses. Moreover, JS-K could overcome the survival and growth advantages conferred by interleukin-6 (IL-6) and insulin-like growth factor-1 (IGF-1), or by adherence of MM cells to bone marrow stromal cells (BMSC). JS-K caused a transient G2/M arrest followed by apoptosis, as determined by flow cytometric analysis using PI, Annexin V and Apo2.7 staining. JS-K-induced apoptosis was associated with caspase 8, 7, 9 and 3 activation. Interestingly, Fas was upregulated by JS-K, suggesting the involvement of death receptor pathway in induction of apoptosis. JS-K also triggered Mcl-1 cleavage and Bcl-2 phosphorylation, suggesting the involvement of mitochondrial pathway. In addition, apoptosis inducing factor (AIF), endonuclease G (EndoG) and cytochrome c were released into the cytosol during apoptosis. Taken together, these findings suggest the involvement of both intrinsic and extrinsic apoptotic pathways in JS-K-induced apoptosis in MM cells. In summary, our studies demonstrate that JS-K induces apoptosis and overcomes in vitro drug resistance in MM cells. Therefore, JS-K is a novel compound which carries significant potential to be included in the repertoire of existing treatment modalities for MM. Ongoing studies are delineating the mechanism of action of JS-K to provide the preclinical rationale for combination therapies to overcome drug resistance and improve patient outcome.
Acceleration of Leukemogenesis in NUP98-HOXD13 Transgenic Mice by Infection with the MOL4070LTR Virus, and Identification of Collaborating Genes.
Authors: Slape Christopher , Lin Yingwei , Bies Juraj , Wolff Linda , Aplan Peter D. 
Published: April 2020

Abstract
NUP98 is a promiscuous oncogene that is fused to at least sixteen different partner genes in a wide range of haematopoietic malignancies. The NUP98-HOXD13 (NHD13) fusion gene is formed by the t(2;11)(q31;p15), which has been observed in patients with MDS or AML. This fusion gene encodes a protein that fuses the N-terminal portion of NUP98, a nucleoporin involved in mediating RNA and protein transport in and out of the nucleus, with the C-terminal portion of HOXD13, a homeodomain protein not expressed during normal haematopoietic development. We have previously demonstrated that expression of the NHD13 fusion gene in a transgenic mouse model results in an invariably fatal myelodysplastic syndrome (MDS). The median survival for these transgenic mice is 10 months, with no mice surviving beyond 14 months of age. About one-third die due to complications of severe pancytopenia, and about two thirds progress to acute leukemia. To identify genes that might collaborate with the NHD13 transgene, a cohort of thirty mice (15 NHD13 transgenic, 15 normal) were infected with the MOL4070LTR virus at birth and the disease course monitored. All transgenic infected mice developed an acute leukemia, predominantly of the myeloid lineage. The median survival of the transgenic infected mice was only four months, with none surviving beyond 7 months of age. These survival figures are vastly decreased compared to either control group (the wild type infected group or the transgenic uninfected group), suggesting a true synergistic effect between the NUP98-HOXD13 transgene and the genes affected by retroviral insertion events. Cloning of these insertion sites has revealed proximity to numerous genes of interest, including several genes with a known role in haematopoiesis (EpoR, Ifnar2, Gata2, PU.1), development (HOXB8, HOXC9) and/or cancer (p53, Stat5b, Pim1, CyclinD1). In addition two recurrent integration sites were identified near uncharacterized ESTs. Since the NHD13 fusion has been shown to block differentiation, several of the genes we identified (p53, EpoR, Stat5b, Pim1, CyclinD1) fit the evolving AML paradigm that postulates one mutation blocks differentiation, and a second, complementary mutation leads to increased proliferation or decreased apoptosis. Ongoing analysis of these and additional collaborating genes should provide important insights into the process of the MDS to AML transition.