bluuebunny/crossref_metadata_2025_split huggingface dataset:
JS-K, a GST-Activated Nitric Oxide Generator, Induces DNA-Double Strand Breaks and Inhibits Growth and Survival of Multiple Myeloma Cells In Vitro and In Vivo.
Authors: Kiziltepe Tanyel , Hideshima Teru , Raje Noopur , Ishitsuka Kenji , Ocio Enrique M. , Catley Laurence , Li Chun-Qi , Trudel Laura , Yasui Hiroshi , Shirashi Norihiko , Tai Yu-Tzu , Chauhan Dharminder , Mitsiades Constantine , Saavedra Joseph E. , Wogan Gerald N. , Keefer Larry K. , Shami Paul J. , Anderson Kenneth C. 
Published: October 2019

Abstract
JS-K (O2-(2,4-dinitrophenyl) 1-[(4-ethoxycarbonyl)piperazin-1-yl]diazen-1-ium-1,2-diolate) is a diazeniumdiolate class of prodrug which is designed to release nitric oxide (NO·) on reaction with glutathione S-transferases (GST). GST has been shown to be overexpressed in a broad spectrum of tumor cells. Therefore, JS-K can possibly turn GST overexpression to the tumor’s disadvantage by generating high intracellular concentrations of cytotoxic NO·. Multiple myeloma (MM) is currently an incurable hematological malignancy where new treatment options are urgently needed. In this study we investigated the cytotoxicity of JS-K in MM in vitro and in vivo. JS-K showed significant cytotoxicity in both conventional therapy-sensitive and -resistant MM cell lines, as well as patient MM cells (IC50: 0.3–2.5 mM). Importantly, no significant cytotoxic effects of JS-K at these doses were observed in normal peripheral blood mononuclear cells. JS-K treatment induced apoptosis in MM cells which was associated with PARP, caspase 8, and caspase 9 cleavage; increased cell surface expression of Fas/CD95; Mcl-1 cleavage; Bcl-2 phosphorylation; as well as mitochondrial cyt c, AIF, and EndoG release. Moreover, JS-K could overcome the survival and growth advantages conferred by exogenous IL-6 and IGF-1, or by adherence of MM cells to bone marrow stromal cells. Flow cytometry experiments revealed significant NO· generation in JS-K-treated MM cells. Since NO· is known to cause DNA double strand breaks (DSB), we hypothesized that JS-K induces DSB in MM cells and confirmed DSB formation by neutral comet assay. We further showed that JS-K also activated DNA damage response pathways as evidenced by H2AX, Chk2 and p53 phosphorylation. In addition, JNK was also activated by JS-K treatment in MM cells, and inhibition of JNK significantly decreased JS-K-induced cytotoxicity, suggesting that JS-K induced apoptosis is mediated via JNK signaling. Finally, JS-K was also significantly effective in inhibiting tumor growth and prolonging median survival (p < 0.01) in a human plasmacytoma xenograft mouse model. Analysis of tumors harvested from treated animals showed that JS-K induced apoptosis and decreased angiogenesis in vivo. Taken together, these data provide the preclinical rationale for the clinical evaluation of JS-K to improve patient outcome in MM.
Mi-63, a Small Molecule Inhibitor of MDM2-p53 Interaction, Has Significant In Vitro Activity in Multiple Myeloma.
Authors: Kumar Shaji , Timm Michael , Kline Michael P. , Haug Jessica L. , Kimlinger Teresa K. , Yang Dajun , Rajkumar S. Vincent 
Published: October 2019

Abstract
Background: Multiple myeloma (MM) is a plasma cell proliferative disorder that results in considerable morbidity and mortality. As it is incurable with the current therapeutic approaches, more effective therapies based on better understanding of the pathobiology of the disease are needed. In MM, malignant plasma cells are characterized by low proliferative and apoptotic rates compared to other malignancies. The tumor suppressor gene p53, responsible for induction of cellular apoptosis in response to genotoxic stimuli, is relatively intact in most cases of myeloma. However, p53 mutations or deletion can occur late in the course of disease. Here we evaluate a novel small molecule inhibitor of the interaction between p53 and its negative regulator, MDM2, in the setting of myeloma.
Methods and Results: Mi-63 was cytotoxic to several different myeloma cell lines with a median effect observed at approximately 2.5 μM in cell lines including MM1.S that express wild type p53 and between 10–15 μM in cells with mutated p53 as measured using an MTT cell viability assay. Additionally, Mi63 induced cytotoxicity in myeloma cell lines resistant to conventional agents such as Melphalan (LR50), Doxorubicin (Dox40) and Dexamethasone (MM1.R), indicating non-overlapping mechanisms. To evaluate the ability of the drug to induce cell death in the tumor microenvironment, MM cells were co-cultured with marrow stromal cells or in the presence of VEGF or IL-6, two cytokines known to be important for myeloma growth and survival. Mi63 was cytotoxic to myeloma cells under these conditions as well, at doses similar to those seen with myeloma cells alone. Mi63 was able to inhibit proliferation and induce apoptosis in myeloma cells in a dose- and time-dependent fashion, as demonstrated by flow cytometry using Annexin/PI staining as well as cell cycle studies. Treatment of myeloma cells with Mi63 was associated with early mitochondrial membrane depolarization, inversion of Bax/Bcl-2 ratio, and down regulation of Mcl-1, indicating induction of mitochondrial mechanisms of cell death. Mi63 was also cytotoxic to freshly isolated primary patient myeloma cells, inducing apoptosis in a dose-dependent manner. In the patient cells the drug appears to have a differential effect on the CD45 positive and negative cells.
Conclusion: Mi-63 has significant activity in vitro in the setting of myeloma as demonstrated by its effect on myeloma cell lines and primary patient cells. It clearly induces apoptosis in myeloma cells, with higher activity seen in cells with wild type p53. Given the lack of p53 abnormalities in most of the patients with myeloma, this drug alone or in combination is likely to have significant clinical activity. Studies combining this with various DNA damaging drugs are in progress. These studies will eventually form the framework for future clinical studies.
Molecular Prognosis in Multiple Myeloma: The IFM Experience.
Authors: Decaux Olivier , Magrangeas Florence , Lode Laurence , Charbonnel Catherine , Gouraud Wilfried , Moreau Philippe , Harousseau Jean-Luc , Attal Michel , Bataille Regis , Campion Loic , Loiseau Herve Avet , Minvielle Stephane 
Published: October 2019

Abstract
Multiple myeloma is the second most common hematological malignancy and remains incurable despite high dose chemotherapy. Patients experience survival periods of few months to more than ten years. We used gene-expression profiles of malignant plasma cells obtained at diagnosis to develop and validate biomarkers to predict disease prognosis. Gene-expression profiling was realized on 250 bone marrow samples obtained from patients less than 66 years, treated according to the Intergroupe Francophone du Myelome (IFM) 99 clinical trials, based on double intensive treatments. The median follow-up time was 3 years and 55 patients died from their disease during this period. Supervised methods were performed to identify expression signatures associated with survival in a training group of 182 patients. From these genes, a classifier of prognosis was developed in the training group and validated in an independent validation group of 68 patients. Univariate Cox proportional hazards model at a significant level of p<0.05 and selection procedure from BRB ArrayTools at a significant level of p<0.001 were used to select 50 genes associated with survival. Resampling and permutation tests were performed in an rigorous internal validation procedure and 28 genes with a permutation p<0.005 and a resampling mean p<0.01 were retained. To reduce gene list instability, multiple training/test partitions (100) were performed and the 15 most stable genes were used to build a survival predictor. For each patient, a score based on the first component of principal component analysis was calculated. The patients were ranked according to their score and divided according to the median (low-risk and high-risk). The model was highly significantly associated with survival in the training group (p<0.001) and it was confirmed in the validation group (p<0.001). The relative risk of death for high-risk patients was 8.46 (95% confidence interval (CI) = 3.3 to 21.6) in the training group and was 15.3 (95% CI = 2 to 116.5) in the validation group. Various biological variables including serum beta2-microglobulin, serum albumin, combined in the International Staging System (ISS), the deletions/monosomy of chromosome 13 (del13) and the t(4;14)(p16;q32) have emerged as relevant prognostic factors in MM. The 15-gene prognostic classifier was independent of ISS (p<0.001) with an adjusted hazard ratio of death =7.5 (95% CI = 3.09 to 18.2), of del13 (p<0.001) with an adjusted hazard ratio of death =8.8 (95% CI = 3.7 to 21), and of t(4;14) (p<0.001) with an adjusted hazard ratio of death =9.3 (95% CI = 3.9 to 22.1). These results demonstrate that this molecular predictor is very powerful to identify different categories of patients based on expected survival, and thus may be clinically useful to design therapeutic trials.
Figure Figure
CCR1 Inhibition Impairs Osteoclast Activity and Interaction with Myeloma Cells.
Authors: Vallet Sonia , Raje Noopur , Ishitsuka Kenji , Hideshima Teru , Podar Klaus , Kizeltepe Tanyel , Yasui Hiroshi , Chhetri Shweta , Ocio Enrique M. , Jin Janice , Okawa Yutaka , Ikeda Hiroshi , Boccadoro Mario , Anderson Kenneth C. 
Published: October 2019

Abstract
Multiple Myeloma (MM) is characterized by increased osteoclasts (OC) activity leading to severe bone disease. Several inducers of OC number and activity have been identified, including several key chemokines. In particular, macrophage inflammatory protein-1α (MIP-1α) and RANTES activate CCR1 and CCR5, resulting in increased osteoclastogenesis (Oba et al. Exp Hematol. 2005) and increased OC motility (Yu X. J Bone Miner Res. 2004) These studies underscore the role of CCR1 in promoting osteoclastogenesis. Here we demonstrate the effects of inhibition of CCR1 with a specific receptor antagonist (MLN3897, Millenium Pharmaceuticals) on normal mature OC. Mature OC were obtained by stimulating PBMC of healthy donors with RANKL and MCSF (50 ng/ml) for three weeks. OC expressed high levels of CCR1 and secreted both MIP-1α (1 ng/ml +/− 1.8) and RANTES (12 pg/ml +/− 0.66), suggesting an autocrine effect of these chemokines. Analyzing the bone resorptive ability with a pit formation assay, we observed that MLN3897 impaired OC function in a dose-dependent fashion (at 10 nM: 20% of reduction in resorbed area, at 100 nM: 50% of reduction). We then studied the effects of CCR1 inhibition on OC viability by analyzing nuclear integrity with Hoechst33258 staining. After 12 hours of cytokine-deprivation, MLN3897 enhanced OC nuclear fragmentation and condensation, suggesting a role for CCR1 ligands in OC survival. We next studied the interactions between OC and MM cells. Because OC secrete several chemoattractants, we studied their ability to stimulate RPMI8226 MM cell migration. We first confirmed high expression levels of CCR1 on RPMI8226 MM cells by flow cytometry (86% compared to isotype control). Inhibition of CCR1 with MLN3897 did not induce any direct cytotoxicity or growth arrest of these cells. We then studied the effects of OC on RPMI8226 MM cells. We observed that OC potently stimulated migration of RPMI8226 MM cells (8-fold increase), which was almost completely blocked by treatment with MLN3897. In contrast, MIP-1α alone induced only a modest effect on migration: the highest effective concentration (0.5 ng/ml) induced only a 2-fold increase. Neutralizing MIP-1α antibody partially reversed these effects, suggesting that other factors may contribute to this migratory effect. Our data therefore demonstrate that CCR1 inhibition by MLN3897 impairs mature OC survival and activity. Although MLN3897 does not have any direct antiMM activity, it affects MM-OC interactions and inhibits MM cell migration to OC. Ongoing studies are further characterizing the effects of CCR1 inhibition on MM-OC interactions in order to provide the framework for its clinical evaluation in MM.
Differentiation of Multiple Myeloma and MGUS by Cross Validated Differential Proteomic Analysis.
Authors: Brown Ross D. , Belov Larissa , dos Remedios Cris , Sze Daniel , Cooper Simon , Dolotin Mila , Gibson John , Joshua Douglas E. 
Published: October 2019

Abstract
The recent development of a standardised proteomic microarray technique, DotScan, has allowed an innovative approach to the investigation of haematological disorders. In this study, mononuclear cells from 49 peripheral blood samples were studied to determine whether the technology could identify a differential consensus pattern of antigen expression for patients with monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM). An automated reader simultaneously determined the expression of 82 different cell surface antigens by binding cells to antibodies on microscopic dots on the nitrocellulose-coated region of a microscope slide. The figure illustrates the image from a typical slide. A consensus pattern of antigen expression was analysed in duplicate by cross-validated discriminant analysis and a pilot database of disease groups was accumulated. Cross validated discriminant analysis successfully predicted patients with monoclonal gammopathy (98% success) and a discrete mosaic pattern could differentiate patients with MM who were treated with thalidomide (n=9). As expected, no single antigen could be used to discriminate between multiple myeloma (n=24), MGUS (n=14) and normal controls (n=11). Antigens with the highest ranking for differentiating the monoclonal gammopathies were CD25 (reduced after thalidomide), CD8 and CD57 (high in MM), CD28 (reduced in MM) and CD95 (reduced in MGUS) reinforcing the importance of immunomodulatory mechanisms in both MM and MGUS. Traditional flow cytometry was used to confirm these specific observations but also to demonstrate that the reduced CD28 expression was specific for CD8+ cells and the reduced CD95 expression was on CD8+ CD57+ cells. Three patients with MGUS were misclassified as MM but on review these 3 patients could have been classified as smoldering myeloma. There was no significant difference in the mosaic of 5 long term survivors of MM (> 10 years). This study has demonstrated the potential of using disease-specific databases to compare the mosaic of antigen expression for the diagnosis of monoclonal gammopathies. Long term studies will be required to accurately determine the prognostic significance at diagnosis and the ability of consensus patterns to identify which MGUS patients develop MM.
Figure Figure Specificity and sensitivity of proteomic array for monoclonal gammopathies MGUS MM MM Thal Normal Sensitivity % Specificity % MGUS prediction 5 3 2 4 35 100 MM prediction 0 15 0 0 100 91 MM Thal prediction 0 0 9 0 100 95 Normal prediction 0 0 0 11 100 89
Distinct Roles of Integrins α6 and α4 in Fetal Liver Hematopoietic Stem and Progenitor Cell Homing.
Authors: Qian Hong , Georges-Labouesse Elisabeth , Nyström Alexander , Domogatskaya Anna , Tryggvason Karl , Jacobsen Sten Eirik W. , Ekblom Marja 
Published: October 2019

Abstract
During development and after transplantation, intravenously injected hematopoietic stem and progenitor cells (HSPCs) selectively transmigrate through the sinusoidal walls into the bone marrow (BM) niches to engraft and reconstitute hematopoiesis. The rate of reconstitution following transplantation varies depending on the source of hematopoietic stem cells (HSCs) (To, et al 1992). However, the molecular pathways that control the homing of HSCs, in particular, of fetal HSCs are still not well understood. In the present study we studied the contribution of α6 and α4 integrins in homing of fetal liver HSPCs into adult BM by using function-blocking antibodies and an integrin α6 knockout mouse model. We found an ubiquitous expression of both integrin α6 and α4 receptors on fetal liver Lin−Sca-1+c-kit+ (LSK) HSPCs. Genetic ablation of integrin α6 resulted in reduced homing of fetal liver progenitors (HPCs) to BM of lethally irradiated adult recipients. In agreement with this, the integrin α6 antibody inhibited homing of fetal liver HPCs into BM and spleen. The role of integrin a6 in homing and engraftment of fetal liver HSCs was studied by a competitive repopulation assay by using integrin α6−/− or α6+/+ fetal liver cells. Absence of α6 integrin in fetal liver cells did not cause any engraftment defect or mobilization hypersensitivity as compared to wild-type cells. In agreement with this, anti-integrin α6 antibody did not either inhibit BM homing of short-term or long-term HSCs. In contrast, homing of fetal liver HSCs and HPCs to BM was virtually abrogated after treatment with integrin α4 antibody. Our results show that the α6 integrin receptors are functional during homing of fetal liver HPCs, but not multilineage repopulating HSC in vivo. Furthermore, we show the critical role of integrin α4 receptor for homing of both fetal liver HPCs and multilineage repopulating HSCs to BM, indicating distinct developmentally regulated functions for integrin α6 and α4 receptors during fetal hematopoiesis
Survival Outcomes of Patients Receiving Thalidomide-Dexamethasone for Previously Untreated Multiple Myeloma.
Authors: Thomas Sheeba K. , Giralt Sergio A. , Wang Michael , Delasalle Kay B. , Gavino Maria , Rankin Kim , Alexanian Raymond , Weber Donna M. 
Published: October 2019

Abstract
Background: In three recent Phase 2 and 3 studies, the combination of thalidomide-dexamethasone (TD) induced responses in 64–72% of patients, including complete responses in 4–16% of patients. However, long term outcomes of patients receiving induction therapy with TD have not been well described.
Patients and Methods: In a retrospective fashion, we updated our experience (median f/u of 50 mos.) of 58 patients treated between 7/2000 and 5/2002, with thalidomide at 100mg nightly, increasing by 100mg every 7 days as tolerated to a maximum dose of 400mg (median 200mg), and dexamethasone (20mg/m2) on days 1–4, 9–12, and 17–20 (decreased to days 1–4 only, beginning with cycle 3). Cycles were repeated on day 28 and patients were treated on protocol for a maximum of 4 courses. To prevent thrombosis, 34 patients received therapeutic doses of either warfarin (INR 2–3) or low molecular weight heparin.
Results: Of 58 patients, 31 proceeded to myeloablative therapy followed by autologous stem cell support. Another 16 patients received maintenance chemotherapy until disease progression. The median age at enrollment was 60.5 (37–78), and 60% were male. By International Staging System criteria, 41% of patients were stage 1, 33% stage 2 and 26% stage 3. The overall response rate was 72%, including 12% with CR. Median overall survival (OS) of those who received autologous stem cell transplant (ASCT) exceeds 59.6 months. Median OS of those who did not receive ASCT was 31.8 months. The observed difference in OS may be partially explained by differences in performance status between the 2 groups.
Conclusion: Our survival data with TD induction does not differ significantly from either historical data with alkylating agent based therapy, or from previously noted prolongation of OS after intensive therapy with stem cell support (IFM-90). Further study of the impact of thalidomide and other novel agents as induction therapy in both the ASCT and non-ASCT settings is warranted.
Transvection Mediated by the Translocated Cyclin D1 Locus in B Cell Malignancies.
Authors: Epner Elliot , Huang Jing , Liu Hui , Wang Jin , Fleming W.H. 
Published: October 2019

Abstract
Translocations involving the immunoglobulin heavy chain gene locus (IgH) are common in B cell malignancies. One common target gene is cyclin D1, which is deregulated in most patients with mantle cell lymphoma (MCL) and 20–30% of patients with multiple myeloma (MM). The cyclin D1 locus represents a model locus to study the role of epigenetics in B cell malignancy. Cyclin D1 is known not to be expressed in normal lymphocytes, where the locus is DNA hypomethylated. In genetic variants that have lost the translocated chromosome, the cyclin D1 locus assumes a densely methylated configuration. Reintroduction of the translocated chromosome by somatic cell fusion reverts the methylated normal locus to a hypomethylated state in trans. Thus, the translocated chromosome exerts a trans DNA hypomethylating effect on the normal chromosome that resembles transvection in Drosophila. This is the first evidence of transvection in a human malignancy. Similar to transvection in Drosophila and paramutation in plants, evidence for pairing of the cyclin D1 loci and the participation of small RNA’s in this process has been obtained. Combined FISH/ immunofluorescent studies have shown the colocalization of both the normal and translocated chromosome 11’s at the nucleolus. Using chromatin immunoprecipitation assays, we show that CTCF and nucleophosmin are present at both the normal and translocated chromosomes in B cells expressing cyclin D1 but not in B lymphocytes. Tethering of the translocated and nontranslocated cyclin D1 loci by CTCF/nucleophosmin at the nucleolus provides physical and spatial juxtaposition of the cyclin D1 loci permissive for transvection.
Erythropoietin Therapy and Venous Thromboembolic Events in Patients with Multiple Myeloma Receiving Chemotherapy with or without Thalidomide.
Authors: Zangari Maurizio , Cavallo Federica , Prasad Keshava , Fink Louis , Coon Sharon , Barlogie Bart , Tricot Guido 
Published: October 2019

Abstract
Background: The association between the use of recombinant human erythropoietin (rHUEPO) and venous thromboembolic events (VTE) has been described in cancer patients treated with chemotherapy. A higher risk of VTE has also been observed with the concurrent use of erythropoiesis-stimulating agents and chemotherapy.
Methods: We retrospectively analyzed VTE incidence and concurrent use of erythropoiesis-stimulating agents in patients with multiple myeloma (MM) enrolled in our Total Therapy 2 (TT2) study. TT2 included 4 monthly cycles of induction chemotherapy followed by tandem transplant. Patients were randomly assigned to receive Thalidomide or not during the whole treatment. Both arms otherwise received identical chemotherapy.
Results: The charts of 599 of a total of 668 patients enrolled (90%) were reviewed; 284 patients received Thalidomide, 315 did not. With median age was 57 years, 59% were male, 24% had IgA myeloma. 62% of patients received erythropoietin (erythropoietin alpha 40000 U SQ/week) during the first 4 months of treatment. In thalidomide treated patients, VTE occurred in 33 out of 166 (20%) patients who concomitantly received administration of erythropoietic agents and in 38 out of 118 (32%) patients who did not receive erythropoietin (p=.025). Among patients not receiving thalidomide, VTE occurred in 18 out of 203 (8%) patients who received concomitant administration of erythropoietic agents and in 17 out of 112 (15%) patients who did not receive erythropoietin (p=.10).
Conclusions: The development of thrombosis in patients enrolled on TTII protocol was not increased by the concomitant use of erythropoiesis-stimulating agents and thalidomide. Similarly, erythropoiesis-stimulating therapy did not affect VTE incidence in newly diagnosed myeloma patients treated with intensive chemotherapy without thalidomide.