bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prognostic Analysis of Refractory Anemia in Adult Patients with Myelodysplasric Syndrome in China.
Authors: Wang Xiao-qin , Chen Zi-xing , Chen Shu-chang , Lin Guo-wei , Ji Mei-rong , Liang Jian-ying , Liu Dan-dan , Li De-gao , Ma Yan 
Published: April 2020

Abstract
Several reports indicated a striking difference in age, chromosome abnormalities and prognosis between Western and Eastern MDS patients, even in the same subtype. The prognostic factors in Chinese MDS patients remain unclear. To investigate the prognostic factors and survival rate in Chinese cases with refractory anemia (RA) in adult MDS based on FAB classification, and to evaluate the applicability of international prognostic scoring system (IPSS) for Chinese MDS-RA patients by comparing with the clinical features of Western cases, Three hundred and seven MDS-RA cases were registered and followed-up in Shanghai, Suzhou and Beijing from 1995 to 2006. The longest follow-up duration was 103 months, and the median follow-up duration was 26.7 months. Kaplan-Meier curve, Log-rank and COX regression model were used to analyze the prognostic factors and survival rate. The median age of 307 MDS-RA cases was 52 years. The frequency of 2 or 3 lineage cytopenias was 85.6%. Abnormal karyotype occurred in 35.7% of all MDS-RA patients. By IPSS cytogenetic risk groups, 165 cases (70.2%) were in the good IPSS cytogenetic subgroup, 44 cases (18.7%) in the intermediate IPSS cytogenetic subgroup and 26 cases (11.1%) in the poor IPSS cytogenetic subgroup. According to IPSS, 20 cases (8.5%) were categorized as low risk, 195 cases (83.0%) as intermediate–I risk and 20 cases (8.5%) as intermediate–II risk. The 1–year, 2–year, 3–year, 4–year and 5–year survival rate were 90.8%, 85.7%, 82.9%, 74.9% and 71.2%, respectively. Fifteen cases (4.9%) transformed to acute myeloid leukemia and the median transformation time was 15.9 months (range 3–102 months). Lower white blood cells count (&lt;1.5 × 109/L), platelet count (&lt;30 × 109/L) and cytogenetic abnormalities were identified as independent prognostic factors by multivariate analysis, while the age (≥65 years), hemoglobin level(&lt;60g/L), IPSS cytogenetic subgroup and IPSS risk subgroup were not independent prognostic factors associated with survival time by COX regression analysis. In summary, Chinese patients were younger and had lower incidence of cytogenetic abnormalities and more severe cytopenias than that of the Western patients. However, the Chinese patients have more favorable prognosis than Western patients do. WBC count, platelet count and karyotype are major prognostic factors for prediction of survival and can be helpful in identifying patients with different prognosis and consequently designing the optimal therapeutic strategies. The present IPSS scores based on conditions of western patients may not be perfectly applicable to MDS-RA patients in Asia and need to be modified.
RNAi Screen of the Druggable Genome To Identify Modulators of Bortezomib in Myeloma.
Authors: Zhu Yuan Xiao , Yin Holly , Que Qiang , Tiedemann Rodger E. , Chng Wee-Joo , Shi Chang Xin , Mousses Spyro , Stewart Keith 
Published: April 2020

Abstract
Bortezomib and related proteasome inhibitors have profound pre-clinical and clinical activity in Multiple Myeloma (MM). However, the critical molecular target(s) modulated by proteosome inhibition remains unknown. The overall response rate to bortezomib in relapsed and refractory MM patients is consistently 35%–50%, suggesting specific molecular targets or resistance mechanisms are inherent in patients. None of the known effects of bortezomib has been shown to correlate with clinical responses in patients or to explain the unique sensitivity of MM (as opposed to other malignancies) to these reagents. Therefore, in this study, we have conducted a small interfering RNA (siRNA) screen of the druggable genome in human myeloma cells to identify genes that modulate sensitivity to bortezomib. KMS11 human myeloma cells were reverse transfected with 13,984 synthetic short interfering RNAs (siRNA) against each of 6992 druggable targets (2 siRNA/gene with a single-siRNA per well/384 well format). At 24 hours post transfection, bortezomib was added at IC10, 30, 60 and 90. Cell viability was measured by ATP-dependent luminescence assay 72 hours post drug treatment. The resulting data from primary screening was evaluated for multiple quality control metrics and found to exceed all expected performance parameters with &gt;99% global transfection efficiency, &lt;0.25 CV values across the dataset with minimal plate-to-plate and set-to-set variation observed. From analysis of the primary screening data, a core panel of 320 (4.6% of druggable targets in library) bortezomib sensitizers were identified. 187 (58%) of these were further confirmed in a secondary screening experiment by multiple (2 to 4) independent siRNA. Among these 187 genes (2.6% of the genes investigated), some known targets, such as proteasome subunits are evident. Other genes belonging to the same functional gene families (e.g. TNF receptor family, Ras family, kinases, oncogenes and transporter proteins) or functioning within the same network of signal transduction (e.g. NF-kB, Wnt and estrogen receptor pathways) are evident. Our primary screen also identified genes that attenuate bortezomib-induced cytotoxicity. Members in this group include genes implicated in the negative regulation of cell proliferation and survival, such as NF-kB inhibitors, JNK pathway activators, tumor suppressors, apoptosis inducers and phosphatases. These studies enable dissection of the molecular mechanism of bortezomib in myeloma cells and may serve to point to biomarkers for response prediction and novel chemotherapeutic combination therapies.
Restricted Oligo-Clonal Usage of Monoclonal Immunoglobulin λ Light Chain Germline in POEMS Syndrome.
Authors: Nakaseko Chiaki , Abe Daijiro , Takeuchi Masahiro , Takeda Yusuke , Tanaka Hiroaki , Oda Kayo , Ohwada Chikako , Ozawa Shinichi , Sakaida Emiko , Shimizu Naomi , Masuda Shinichi , Cho Ryuko , Nishimura Miki , Saito Yasushi , Misawa Sonoko , Kuwabara Satoshi 
Published: April 2020

Abstract
Background: POEMS syndrome (also known as Crow-Fukase or Takatsuki syndrome) is a rare plasma cell disorder characterized by peripheral neuropathy, a monoclonal plasma cell disorder, and other paraneoplastic features including organomegaly, endocrinopathy, skin changes, edema and effusions. High levels of vascular endothelial growth factor (VEGF) in the serum may contribute to pathology seen in this disease but exact pathogenetic mechanisms are still unclear. A unique feature of this syndrome is that the proliferating monoclonal plasma cells are virtually always λ-restricted. Here, we determined complete nucleotide sequences of monoclonal immunoglobulin λ light chain (IGL) variable regions in 10 patients with POEMS syndrome and found that Vλ germline usage is highly restricted to the Vλ1 family and oligoclonal germlines.
Materials and methods: Total RNA was extracted from bone marrow mononuclear cells of patients with POEMS syndrome and the V-J region of the IGL gene was amplified by RT-PCR using 5′ degenerative primers for the Vλ consensus leader lesion (5′-ATGGCCKGSWYYSYTCTCCTC-3′) and 3′ primers matching the consensus upstream part of the cl exon (5′-CTCCCGGGTAGAGAAGTCACT-3′). Monoclonal bands were detected by heteroduplex analysis or TA cloning and subjected to nucleotide sequencing. Sequence data were analyzed using the International ImMunoGeneTics information system (IMGT, http://imgt.cines.fr) and mutations were identified by comparison with the germline sequence databases.
Results: Of 13 POEMS syndrome patients with λ-type M protein, nucleotide sequencing of the IGL gene was successful in 10. The V-region of the IGL gene of all 10 patients was restricted to the Vλ1 family. Searching for homologies with IGL germlines revealed that only two were used, with an average homology of 90.1%. IGLV1-40*01 was used in 8 patients and IGLV1-44*01 in the remaining two. In the IGL-J region, the IGLJ3 gene was found in 9 of 10 patients and IGLJ2 in one (10%). All IGLV-J rearrangements were mutated with homologies to the germline sequence ranging from 77.66% to 96.10% (average homology: 90.1%). All CDR3 were composed of 11 amino acids, with identical acidic isoelectric point (pI) values (13.0) and similar molecular weights ranging from 1394.7 to 1552.8 (median, 1506.2).
Conclusions: IGL-M protein in POEMS syndrome belongs to the Vλ1 family and is markedly restricted to specific oligoclonal germlines. Our results suggest that the restricted use of IGL plays an important role in the pathogenesis of POEMS syndrome. These data provide an important insight for elucidating the pathogenesis of POEMS syndrome. To the best of our knowledge, this the first report of restricted oligo-clonal usage of monoclonal immunoglobulin germlines in plasma cell disorders.
ß-Catenin siRNA Successfully Suppressed the Progression of Multiple Myeloma in In Vivo Mouse Model.
Authors: Kawata Eri , Ashihara Eishi , Tanaka Ruriko , Yokota Asumi , Murotani Yoshihide , Taniguchi Miki , Kamitsuji Yuri , Kuroda Junya , Inaba Tohru , Shimazaki Chihiro , Kimura Shinya , Maekawa Taira 
Published: April 2020

Abstract
ß-catenin is the downstream effector of the Wnt signaling pathway and has cyclin D1 and c-myc as its target genes. Activation of Wnt signaling is greatly involved in the process of carcinogenesis, and its overexpression has reported to be positively correlated with prognosis in many types of malignant tumors, as colorectal cancer and hepatocellular carcinoma, or hematological malignancy, as acute myeloid leukemia and chronic myeloid leukemia. Multiple myeloma (MM) is a relatively common hematological malignancy and remains incurable with conventional treatments. Here, we assessed the expression of ß-catenin in MM and growth inhibitory effect of ß-catenin siRNA on MM. We first examined the expression of ß-catenin on human MM cell lines: AMO-1, EJM, IM-9, KMS-12-BM, LP-1, NCIH929, OPM-2, RPMI8226, and U266. All cell lines examined expressed a significantly higher level of ß-catenin than normal human mononuclear cells. Moreover, ß-catenin was also overexpressed in myeloma cells from patients. We then assessed the in vivo growth inhibitory effects of ß-catenin siRNA in mouse model. Six -week-old male Balb/c nu/nu mice were subcutaneously inoculated in the right flank with 5 × 106 RPMI8226 myeloma cells in 100μl PBS. Three to four weeks later, when palpable tumors (100 mm3 in diameter) developed, mice (n = 5 for all groups) were treated with either subcutaneous (around tumors) injections of group A: ß-catenin siRNA (2.5μM) / atelocollagen complex (final atelocollagen concentration 0.5%), group B: control siRNA (2.5μM) /atelocollagen complex, group C: ß-catenin siRNA (2.5μM)/PBS, group D: PBS/atelocollagen twice a week for a total of eight injections. Tumor size was measured in two dimensions using a caliper, and tumor volume (mm3) was calculated as a2 × b /2 mm3 (a;minor axis,b;major axis). The mean tumor volume of each group after two weeks treatment was 412.2 mm3 in groupA, 1317.9 mm3 in group B, 2075.9 mm3 in group C, and 1802.3 mm3 in group D, respectively, and the treatment with ß-catenin siRNA (2.5μM) / atelocollagen complex significantly reduced tumor burdens and retarded tumor growth, measured as tumor volumes (p&lt;0.05) (Figure 1). Therefore, ß-catenin siRNA revealed growth inhibitory effects on MM in in vivo mouse model, which strongly indicates that ß-catenin is the attractive novel target in treating MM.
Figure Figure
The Tyrophostin Adaphostin (NSC680410) Inhibits Multiple Myeloma Bone Marrow Angiogenesis In Vitro and In Vivo.
Authors: Podar Klaus , Zhang Jing , Raab Marc S. , Vallet Sonia , Fulciniti Mariateresa , Sattler Martin , Ikeda Yutaka , Chauhan Dharminder , Anderson Kenneth C. 
Published: April 2020

Abstract
Our own and other previous studies demonstrate marked anti-proliferative activity of the tyrophostin adaphostin (NSC680410) in a variety of hematologic malignancies including chronic myelocytic leukemia (CML), chronic lymphcytic leukemia (CLL), acute myelocytic leukemia (AML), and Multiple Myeloma. Here we show that adaphostin (NSC680410), similar to bortezomib, additionally inhibits tumor angiogenesis within the MM bone marrow (BM) microenvironment. This effect is elicited both indirectly by inhibition of VEGF production and secretion in MM cells, as well as directly by abrogation of endothelial cell growth. Specifically, adaphostin triggers marked downregulation of nuclear c-Myc expression in MM cells. Both adaphostin, as well as specific downregulation of c-Myc using siRNA, lead to a decrease in cobalt chloride- induced Hif-1alpha- expression and Hif-1alpha activity, as evidenced by western blot analysis and expression of Hif-1alpha- driven luciferase, respectively. Indeed secretion of the Hif-1alpha target gene VEGF is markedly inhibited in a dose- and time- dependent manner. Importantly, neither knockdown of c-Abl expression nor exogenous overexpression of caspase- cleavage- induced c-Abl fragment abrogates drug- induced Hif-1alpha downregulation or inhibition of its activity. Taken together, these results indicate the existence of a c-Myc/ Hif-1alpha- dependent, but c-Abl- independent, pathway modulating MM cell production and secretion of VEGF. In contrast, we demonstrate a direct antiangiogenic effect of adaphostin on endothelial cells, similar to H2O2, is mediated via c-Jun upregulation, inhibition of cell proliferation, and the induction of cell apoptosis. Moreover, our data further demonstrate activity of adaphostin within the BM microenvironment. Adaphostin, similar to bortezomib, significantly inhibits VEGF secretion triggered by adhesion of MM cells to BMSCs and endothelial cells. Consequently, conditioned medium derived from adaphostin- treated co-cultures markedly inhibits endothelial cell growth and tubule formation in a dose- dependent manner. Finally, we confirmed these in vitro results using an in vivo xenograft mouse model of human MM. Specifically, western blot analysis, as well as immunohistochemistry, demonstrate marked downregulation of both Hif-1alpha and CD31 in tumors isolated from adaphostin- treated animals versus control animals, confirming the in vivo antiangiogenic effect of adaphostin. Similar effects were obtained using a SCIDhu mouse model as well as a significant decrease of MM- related bone disease, due to anti- VEGF activity of adaphostin. Taken together, these data provide the rationale for the clinical evaluation of adaphostin to target both MM cells and the BM milieu to improve patient outcome in Multiple Myeloma.
Activated CMRF-56 Selected Blood Dendritic Cells: A Potential Anti-Myeloma Vaccine.
Authors: Freeman Jennifer L. , Vari Frank , Hart Derek N.J. 
Published: April 2020

Abstract
Despite recent advances in therapy, such as stem cell transplantation and new drugs including proteosome inhibitors, multiple myeloma (MM) remains incurable for most patients. Immune therapy using dendritic cell (DC) vaccination to generate anti-myeloma cytotoxic T cell (CTL) responses targeting residual disease is a promising, potentially curative strategy that would be compatible with other therapies. Blood DC isolated using the monoclonal antibody (mAb) CMRF-56, can induce CTL responses (J. Immunotherapy. 2005; 28:322). We have established the ability of CMRF-56 blood DC to migrate in vitro and induce CTL responses to MM antigens and developed clinical scale production of the CMRF-56 blood DC preparations. Migration of CMRF-56 blood DC toward a CCL21 gradient was significantly improved by addition of granulocyte macrophage colony stimulating factor (GM-CSF) and prostaglandin E2 (PGE2) (n=5, p&lt;0.05) and exceeded that of activated MoDC. Activation of CMRF-56 blood DC with GM-CSF also optimal for induction mature phenotype (n=4, p&lt;0.05), survival of blood DC after 48 hours culture (n=3, p&lt;0.01), and generation of CTL (n=5, p&lt;0.05). Addition of PGE2 made no significant difference to CTL induction. GM-CSF activated CMRF-56 blood DC loaded with U266 lysate induced polyclonal anti-myeloma CTL responses capable of 38–61% specific lysis of U266 cells in a 51Cr release assay. To determine specificity, we examined the CTL specific for known MM tumour antigens and could identify CTL recognising Muc1 and HM1.24 HLA-A*201 epitopes in 3/3 donors tested. CMRF-56 blood DC derived from both healthy and MM donors were pulsed with HLA-A*201 restricted peptides derived from MM tumour antigens including Muc1 and HM1.24/BST2, along with novel antigens such as cancer testis antigen (CT)-7 (MAGE-C1) and B cell maturation antigen (BCMA). Results from healthy donors have shown that CTL specific for all above mentioned antigens can be induced with 4/8 responding to HM1.24, 2/8 to Muc1, 4/8 to CT7 and 1/6 to BCMA and these CTL lysed U266 cells specifically. In parallel, we have developed and validated clinical scale production of CMRF-56 blood DC preparations using clinical grade biotinylated CMRF-56 mAb and the CliniMACS system. On average over 100 million CMRF-56 blood DC were recovered from immunoselected apheresis products from both healthy (n=10) and MM donors (n=4) with average blood DC purities of 31% and 42% respectively. These results indicate that CMRF-56 blood DC can be produced on a clinical scale, can migrate toward secondary lymphoid chemokine CCL21 in vitro and can induce anti-myeloma CTL responses to both known and novel MM antigens. These data supports the use of CMRF-56 blood DC preparations loaded with peptide antigen as a potential anti-myeloma vaccine for patients with minimal residual disease.
The Novel, Orally Available Multi-Kinase Inhibitor BAY 73-4506 Targets Myeloma Cells and Their Bone Marrow Microenvironment.
Authors: Raab Marc S. , Breitkreutz Iris , Klaus Podar , Zhang Jing , Blotta Simona , Christensen Olaf , Hideshima Teru , Chauhan Dharminder , Wilhelm Scott , Anderson Kenneth C. 
Published: April 2020

Abstract
Multitargeted treatment approaches have been shown to be more effective than single agent therapy in multiple myeloma (MM). In addition, agents targeting not only the MM cells directly but also their microenvironment, like bone marrow stromal cells (BMSCs), endothelial cells, and osteoclasts (OCLs) causing enhancement of tumor cell growth, angiogenesis, and MM bone disease, respectively, are promising new treatment modalities for this still non-curable disease.Here we investigated the novel, orally available multi-kinase inhibitor BAY 73-4506, currently in phase I clinical trials, for its therapeutic effect in MM. BAY is a potent inhibitor of angiogenic (VEGFR 1-3, PDGFR-b), as well as oncogenic, kinases (cKIT, RET, FGFR, Raf). We first tested the ability of BAY to suppress MM cell proliferation and survival in a wide array of MM cell lines (MM.1S, RPMI 8226, NCI H929, OPM2, KMS11, KMS 18, INA6, U266, KMS12BM, S6B45), including those resistant to conventional chemotherapeutics (MM.1R, Dox40, LR5). Our data show that BAY is active in all cell lines tested in a low micromolar range equivalent to concentrations achieved in patient plasma during the first clinical trial in solid tumors. Importantly, BAY also overcomes the growth advantage conferred in a BMSC-MM, as well as an endothelial cell-MM, coculture system. BAY treatment abrogates MEK, ERK and AKT phosphorylation in a time and dose dependent manner, followed by induction of apoptosis, evidenced by Annexin staining and DNA fragmentation. Since VEGF signaling pathway is a potent inducer of angiogenesis and BAY targets VEGFR 1-3, we examined anti-angiogenic properties of BAY. This compound inhibits endothelial cell growth and endothelial cell tubuli formation in vitro at concentrations less than 1mM; moreover, BAY markedly inhibits the VEGF-induced cell migration on fibronectin. Activation of MAP kinase is a critical event during OCL differentiation, activation, and survival; BAY inhibits osteoclastogenesis, evidenced by blockade of M-CSF/RANKL-triggered differentiation of mononuclear cells to TRAP-positive osteoclasts, an important marker of osteoclastogenesis. Finally, combination treatment of BAY with dexamethasone shows synergistic effects on MM cell growth and survival. These in vitro experiments on the effects of BAY on MM tumor cells directly, in co-culture with endothelial or BMSCs, as well as on osteoclast differentiation, provides the basis for its evaluation in a murine model of human MM to confirm these promising in vitro effects of this novel multi-kinase inhibitor, finally leading to clinical evaluation to improve patient outcome.
Transgenic Expression of JAK2V617F Causes Myeloproliferative Disorders in Mice.
Authors: Xing Shu , Zhao Zhizhuang Joe 
Published: April 2020

Abstract
Recently, an acquired mutation of tyrosine kinase JAK2 was found in most patients with myeloproliferative disorders (MPDs) including polycythemia vera (PV), essential thrombocythemia (ET), and idiopathic myelofibrosis. We have generated transgenic mice expressing the mutation enzyme, JAK2V617F, in the hematopoietic system driven by the promoter of the vav gene. The mice are viable and fertile. One line of the transgenic mice expressed a lower level of JAK2V617F and displayed elevated blood cell counts, while the other line expressed a higher level of JAK2V617F and exhibited a marked increase in blood counts and developed phenotypes that closely resembled human ET and PV. The latter line of mice also developed marrow and spleen fibrosis as the animal aged. In general, the transgenic mice had megakaryocytic hyperplasia in the bone morrow and extramedullary hematopoiesis resulting in splenomegaly, and their serum erythropoietin level was also significantly reduced. In vitro colony assays demonstrated that transgenic mice possessed an increased number of hematopoietic progenitor cells in peripheral blood, spleen, and bone marrow and that these cells displayed hyper-sensitivity to growth factors and cytokines. The data prove that JAK2V617F is a cause of MPDs. Our study thus provides a permanent mouse system for further study to define the pathological role of JAK2V617F and to develop treatment for MPDs.