bluuebunny/crossref_metadata_2025_split huggingface dataset:
Angiogenic Signature of Multiple Myeloma Determined by Gene Expression Profiling.
Authors: Hose Dirk , Moreaux Jerome , Meissner Tobias , Rossi Jean-Francois , Benner Axel , Mahtouk Karene , Seckinger Anja , Heiss Christiane , Hundemer Michael , Reme Thierry , Hillengass Jens , Herde Katja , Bertsch Uta , DeVos John , Wenisch Sabine , Pantesco Veronique , Jauch Anna , Jourdan Eric , Goldschmidt Hartmut , Klein Bernard , Moehler Thomas 
Published: April 2020

Abstract
BACKGROUND. Angiogenesis is a hallmark of active multiple myeloma and a therapeutic target, e.g. by thalidomide. Others and we have shown this to be due to a differential and novo expression of pro/antiangiogenic genes in MM as well as an effect of increased plasma cell number. AIM of this study was to investigate based on the expression of (anti)angiogenic genes alone (consensus list determined by medline search),whether for respective samples being PPC, BMPC or MMC can be predicted,what expression differences between these entities exist,if differences between the expression pattern of MMC from early stage vs. therapy requiring MM can be used for prediction, andwhether the survival of MM–patients undergoing HDT is different in two groups distinguished by their angiogenic signature.
PATIENTS AND METHODS. 187 newly diagnosed MM/MGUS–patients (65 training (TG) / 122 independent validation group (VG)), 14 normal donors (ND) and 12 in vitro generates PPC samples were included. Bone marrow aspirates were CD138-purified by activated magnetic cell sorting. RNA was in-vitro transcribed and hybridised to Affymetrix HG U133 A+B GeneChip (TG) and HG U133 2.0 plus arrays (VG). Expression data were gcrma-normalised and the empirical Bayes algorithm used. P-Values were adjusted using the Benjamini-Hochberg method (Bioconductor). The PANP-algorithm was used to identify expressed genes. iFISH was performed on purified MM-cells using probesets for chromosomes 1q21, 9q34, 11q23, 11q13, 13q14, 15q22, 17p13, 19q13, 22q11 and the translocations t(4;14) and t(11;14). Selected expression data were verified by real time RT-PCR and western blotting.
RESULTS.
On TG and VG, being BMPC or PPC is predicted by a 17 gene predictor (31 probe sets) without error. Being MMC is predicted with an error rate of 6.9/4.7% with an overall-error rate of 5.8/4.3% in TG/VG, respectively. 5 proangiogenic genes/7 probe sets (HGF [hepatocyte growth factor], ADM [adrenomedullin], CXCL2 [chemokine (C-X-C-motif)-ligand 2], IGF1 [insulin-like growth factor 2], Met [HGF-receptor]) and 1 antiangiogenic/1 probe set (SerpinF1, [Serpin peptidase inhibitor F1]) are differentially expressed between these entities concordantly in TG and VG. Attribution of MMC to early or therapy requiring can not be predicted. Most patients with MGUS and MM display 1 or several over expressed angiogenic gene. Survival analysis (EFS/OAS) will now be performed based on these findings.
CONCLUSION. Our analysis reveals novel targets for antiangiogenic therapy of multiple myeloma as anti-HGF. There is evidence that angiogenesis related genes are activated early during disease progression as rationale for early intervention using antiangiogenic compounds.
Molecular and Transcriptional Characterization of the Novel 17p11.2-p12 Chromosome Amplification in Multiple Myeloma.
Authors: Fabris Sonia , Todoerti Katia , Mosca Laura , Agnelli Luca , Ronchetti Domenica , Intini Daniela , Lionetti Marta , Ciceri Gabriella , Deliliers Giorgio Lambertenghi , Bertoni Francesco , Neri Antonino 
Published: April 2020

Abstract
Multiple myeloma (MM) is a fatal malignancy of clonal bone marrow plasma cells characterized by a high genomic instability increasing with disease progression. We describe here a genomic amplification at 17p11.2–p12, an unstable chromosomal region characterized by a large number of low copy repeats which have been proven to mediate deletion and duplication in several genomic disorders and amplifications in solid tumors. An approximately 5 Mb 17p11.2–p12 amplified region was detected in KMS-26 myeloma cell line by SNP microarray analysis. Further FISH mapping showed two unidentified amplified chromosomes as well as a complex pattern of rearranged chromosomes 17. The analysis of transcriptional profiles in a proprietary data base of myeloma cell lines, identified 12 significantly overexpressed genes in KMS–26 amplified region, including TNFRSF13B/TACI, COPS3, and NCOR1. The evaluation of their expression levels in a data base including 11 monoclonal gammopathy of uncertain significance, 121 MM and 9 plasma cell leukaemia, showed a significant overexpression of at least one gene in 13 patients. FISH analyses of these patients identified one MM carrying a 3.8 Mb amplified region and two MMs with gains specifically involving the TACI locus. Interestingly, the complete inactivation of TP53 at 17p13.1 was found in KMS–26 whereas a monoallelic loss was identifiable in two of the three patients carrying gain/amplification. Our data suggest that, as in the case of solid tumors, amplification/gain of the 17p11.2–p12 in MM could be mediated by the presence of repeats located in this region and may provide insights for defining novel candidate myeloma-associated genes.
Evaluation of the Cytogenetic Aberration Pattern in AL Amyloidosis Compared to Monoclonal Gammopathies Not Requiring Treatment: Translocation t(11;14) Is More Frequent in AL Amyloidosis.
Authors: Bochtler Tilmann , Schonland Stefan O. , Jauch Anna , Heiss Christiane , Benner Axel , Cremer Friedrich W. , Hose Dirk , Moos Marion , Bartram Claus R. , Ho Anthony D. , Goldschmidt Hartmut , Hegenbart Ute 
Published: April 2020

Abstract
Introduction: Cytogenetic aberrations (CA) have emerged as important pathogenetic and prognostic factors in plasma cell disorders. However, in AL amyloidosis (AL) only a few reports with small numbers of patients have been published.
Methods: Using interphase FISH analysis in CD138+ cells, we evaluated the role of CA in a series of 85 AL patients as compared to 146 patients with a monoclonal gammopathy without treatment requirement in a prospective manner. Our panel included IgH translocations t(11;14), t(4;14), t(14;16) and translocation of 14q32 with an unidentified partner, gains of 1q21, 11q23 and 19q13 as well as deletions of 8p21, 13q14 and 17p13. Using these probes we could detect at least one of these aberrations in 95% of AL and in 88% of the control group. Age, gender and plasma cell content were statistically equally distributed among both groups.
Results: The most frequent aberration in AL was t(11;14), which was detected in 45% of AL patients as compared to 26% of the control group (p=0,056). It was strongly associated with the lack of an intact immunoglobulin (p&lt;0,001), thus accounting for the frequent light chain only subtype in AL. Markedly, t(11;14) was more frequently found in combination with gain 11q23 in AL than in the control group (20% versus 6%, p = 0.005). Other frequent aberrations in AL included deletion 13q14 (32%) and gain 1q21 (21%), which were observed in the control group at comparable frequencies (34% and 20%). The overlapping character of the underlying plasma cell disorder in both disease entities was also emphasized by the similarities of branching patterns of the five major CA in cluster analysis applied in 169 patients (figure 1). The relation of clinical parameters and chromosomal aberrations was also evaluated. The analyzed CA had no impact on the organ involvement pattern in AL patients.
Conclusions: We observed a high frequency for t(11;14) in AL. Apart from this finding, the cytogenetic patterns known in monoclonal gammopathy of unknown significance and multiple myeloma were widely shared by AL amyloidosis.
Figure Figure
Cathepsin G, a Potent Platelet Aggregation Activator, Is Upregulated in Multiple Myeloma Patients during Treatment with Lenalidomide (Revlimid).
Authors: Pal Rekha , Kennedy Ryan , Mansour L. Renee , Stirling David , Mapara Markus Y. , Roodman G. David , Lentzsch Suzanne 
Published: April 2020

Abstract
Lenalidomide induces impressive response rates in patients with multiple myeloma. Unfortunately, one of the major side effects of this therapy is the increased risk of thromboembolic events (TEE) up to 17%. The reason for a higher risk of TEE associated with Lenalidomide is not known, but it has been found that aspirin is very effective in preventing TEEs induced by thalidomide and lenalidomide (Rajkumar, 2005). Cathepsin G is a serine proteinase present in the azurophilic granules of polymorphonuclear neutrophils (PMNs) and is a strong platelet aggregation agonist with potency similar to thrombin. In the current study we investigated the effect of the immunomodulatory derivatives of thalidomide (IMiDs) including Actimid and lenalidomide on cathepsin G in vitro and in vivo. To investigate the effect of IMiDs in vitro CD34+ hematopoietic progenitors were cultured in the presence of IL-3, IL-6 and SCF supporting the development of granulocytes and treated with Actimid or DMSO (control). RNA and protein were extracted for the analysis of cathepsin G. A significant up-regulation of cathepsin G (7.7 fold) was detected in oligonucleotide gene array analysis after 3 days of treatment with Actimid as compared to control group. These results were confirmed by RT-PCR, which showed a 3.7 and 7.6 fold mRNA-increase on day 6 and day 10 of culture compared to control. Next we analyzed the cathepsin G levels of patients (n=10) treated with lenalidomide before treatment, on day 15 of each cycle (1–4) as well as 1, 2, and 3 month after stop of lenalidomide treatment. We observed a continuous significant increase of the mRNA levels of PMNs over the course of treatment (baseline: 1 fold, cycle 2: 1.7 fold, cycle 3: 4.8 fold, cycle 4: 20.7 fold). These data were confirmed by measuring cathepsin G levels in patient serum by ELISA. Cathepsin G significantly increased from a baseline mean of 53 ng/ml to 77.5 ng/ml (cycle 2), 129.2 ng/ml (cycle 3) and 145.5 ng/ml (cycle 4). After stop of the lenalidomide treatment we observed a time dependent normalization to baseline levels of the cathepsin G with 61 ng/mL (1 month post treatment), 42.5 ng/mL (2 months post treatment), and 49.2 ng/mL (3 months post treatment). Our data show that IMiDs up-regulate the potent platelet aggregation activator cathepsin G in hematopoietic cells and thereby might contribute to the development of TEE in patients receiving IMiD treatment. Based on our data TEE prophylaxis should be continued for at least 1 more month after stop of therapy. For the first time our results provide an explanation why aspirin is effective in preventing TEE in patients receiving IMiDs. Further studies are needed to determine whether cathepsin G helps to predict TEE. Inhibition of cathepsin G might be a potential therapeutic target for preventing the hypercoaguable state induced by IMiD treatment.
Autophagy as a Target Pathway in Multiple Myeloma: A Forward Chemical Genetic Approach.
Authors: Shen John P. , Divakaran Sanjay , Kuai Letian , Wang Xiang , Ong Shao-En , Amaravadi Ravi , Wood John , Carr Steven , Schreiber Stuart , Haggarty Stephen , Bradner James 
Published: April 2020

Abstract
Autophagy is an evolutionarily conserved process by which cellular structures may be degraded to support ongoing biogenesis. Recent evidence has identified the induction of autophagy following growth factor pathway inhibition, rapid tumor expansion and proteasome inhibition. Thus, autophagy is a highly desirable pathway for the development of targeted therapeutics in cancer, in particular in tumors characterized by misfolded protein stress such as multiple myeloma (MM). Unfortunately, there are only several known classes of autophagy inhibitors and the emerging biology of autophagy has identified few validated targets. We therefore performed a forward chemical genetic study of autophagy using novel techniques in high-content imaging, high-throughput screening and multidimensional data analysis. Using LN229 glioblastoma cells stably transfected with an LC3-EGFP construct, we created a robust screening platform capable of quantifying the number, size and fluorescence intensity of autophagosomes. Assay validation was performed with the known autophagy inhibitor chloroquine, which induces a characteristic shift from diffuse to punctate fluorescence. Initially, we profiled a library of approximately 3,500 structurally diverse bioactive small molecules to leverage possible downstream mechanistic insights. Assay positives and controls were re-tested in the adherent H1299 adenocarcinoma cell line likewise stably expressing LC3-EGFP. Known autophagy inhibitors were enriched with this approach: nocodazole, bafilomycin A1, colchicine and monensin. Structure-activity relationship analysis of the primary screening data and 34 novel autophagy modulators revealed a compelling activity of bis-indolyl maleimides. We confirmed sub-micromolar inhibition of autophagy by three such compounds using a quantitative, radiolabeled protein degradation assay (tritiated tyrosine pulse-chase), LC3-II immunoblotting and quantitative measurement of autophagosomes across three cell lines. From these studies we selected a lead compound, K252a, for further characterization and tested twenty-two closely related analogues. Detailed SAR analysis has identified features of the molecule required for inhibition of autophagy and sites permissive for further functionalization in ongoing target identification studies. Using MM as a translational model system of misfolded protein stress, we found dose-dependent, selective cytotoxicity in MM of K252a compared to analogues lacking autophagy inhibitory activity. We report here the results of a forward chemical genetic study of autophagy which identifies and characterizes novel small molecule inhibitors of autophagy and establishes a rationale for clinical studies of autophagy inhibitors in the treatment of MM.
Wnt3/RhoA/ROCK Signaling Pathway Is Involved in VLA6-Dependent Adhesion-Mediated Drug Resistance of Multiple Myeloma.
Authors: Kobune Masayoshi , Kawano Yutaka , Takimoto Rishu , Matsunaga Takuya , Kato Junji , Hamada Hirofumi , Niitsu Yoshiro 
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 multiple myeloma. 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 β-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, anti- integrinα6 antibody and 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 integrinα6/β1 (VLA-6)-mediated attachment to stromal cells is induced by Wnt/RhoA-Rho kinase (ROCK) pathway signal. This CAM-DR for doxorubicin was also significantly reduced by Wnt3 siRNA transfer to KMS-5 and further augmented by addition of Wnt3 conditioned medium. These results indicate that Wnt3 contributes to VLA-6-mediated CAM-DR via the Wnt/RhoA/ROCK pathway of myeloma cells. Thus, the Wnt3/RhoA/ROCK signaling pathway could be a promising molecular target to overcome CAM-DR.
KD7150, a Novel Mercaptoketone-Based HDAC Inhibitor, Exerts Potent Anti-Multiple Myeloma Effects In Vitro and In Vivo by Induction of DNA Damage and Mitochondrial Signaling.
Authors: Feng Rentian , Ma Huihui , Hassig Christian A. , Payne Joseph E. , Smith Nicholas D. , Mapara Markus Y. , Hager Jeffrey H. , Lentzsch Suzanne 
Published: April 2020

Abstract
Histone deacetylase (HDAC) inhibitors have emerged as a class of novel and promising anti-cancer agents and their activities are currently being investigated in both the pre-clinical and clinical settings. Using an unbiased, ultrahigh throughput screening system a novel mercaptoketone-based HDAC inhibitor KD5170 was identified. This novel non-hydroxamic acid non-benzamide compound was profiled for its anti-myeloma activity in vitro and in vivo. KD5170 inhibited the proliferation of multiple myeloma (MM) cell lines and the viability of CD138+ primary MM cells by induction of apoptosis. This was accompanied by an increase of acetylation of histones and activation of caspase-3, -8 and -9. Treatment with KD5170 caused a loss of mitochondrial membrane potential measured by JC-1 staining and resulted in release of apoptogenic factors such as cytochrome c, smac, and AIF from mitochondria. Furthermore, KD5170 induced oxidative stress and oxidative DNA damage in myeloma cells, as evidenced by the upregulation of heme oxygenase-1 and H2A.X phosphorylation, respectively. Combination of KD5170 with the proteasome inhibitor bortezomib or TNF-related apoptosis-inducing ligand synergistically enhanced the anti-myeloma activity. Resistance of MM cells to KD5170 was associated with activation of the ERK/MAPK pathway under treatment with KD5170. Pretreatment with the MAPK-inhibitor restored the sensitivity to KD5170, suggesting that the combination of KD5170 with a MAPK inhibitor (U0126) could overcome drug resistance. Growth of myeloma tumor xenografts in KD5170-treated nude mice was significantly inhibited (1401 mm3 ± 596 versus 442 mm3 ± 221; P = .009) and survival was prolonged (9 days and 17.5 days; P= .0095) compared to vehicle-treated mice. Accordingly, our in vivo data showed that histone acetylation was markedly upregulated in tela in spleen or tumor tissues of the animals treated with KD5170 as early as 2 hours. In conclusion, our data show that KD5170 has anti-myeloma activity in vitro and in vivo, mediated by the induction of apoptosis due to DNA damage and mitochondrial signaling.
Combination of Proteasome Inhibitors Bortezomib and NPI-0052 Trigger In Vivo Synergistic Cytotoxicity in Multiple Myeloma.
Authors: Chauhan Dharminder , Singh Ajita , Brahmandam Mohan , Podar Klaus , Hideshima Teru , Schlossman Robert , Richardson Paul , Munshi Nikhil , Palladino Michael , Anderson Kenneth C. 
Published: April 2020

Abstract
Bortezomib (Velcade™), the first in class proteasome inhibitor is FDA approved drug for the treatment of relapsed and relapsed/refractory multiple myeloma (MM). However, as with other agents, dose-limiting toxicities and the development of resistance limits its long-term utility. Our recent study demonstrated that a novel proteasome inhibitor NPI-0052 triggers apoptosis in MM cells; and importantly, is distinct from bortezomib in its chemical structure, effects on proteasome activities, and mechanisms of action. Here, we demonstrate that combining NPI-0052 and bortezomb induces synergistic anti-MM activity both in vitro using MM cell lines or patient CD138+ MM cells and in vivo in a human plasmacytoma xenograft mouse model. NPI-0052 + bortezomib-induced synergistic apoptosis is associated with:
activation of caspase-8, caspase-9, caspase-3, and PARP; induction of ER-stress response and JNK; inhibition of migration of MM cells and angiogenesis; suppression of chymotrypsin-like (CT-L), caspase-like (C-L) and trypsin-like (T-L) proteolytic activities; and blockade of NF-kappa B signaling.
Studies in a xenograft MM model show that low dose combination of NPI-0052 and bortezomib is well tolerated and triggers synergistic inhibition of tumor growth. Importantly, analysis of resected xenografted tumors show that 30–40% proteasome inhibition of all three (CT-L, C-L and T-L) proteasomal activities is sufficient to trigger significant MM cell apoptosis, confirming both the sensitivity of MM cells to proteasome inhibition and the importance of inhibiting all three proteolytic activities to obtain maximum response. Immunohistochemical analysis of MM tumors excised from NPI-0052 + bortezomib-treated mice showed growth inhibition, apoptosis, and a decrease in associated angiogenesis. The clinical observation that bortezomib therapy can be associated with toxicity and drug-resistance, coupled with our present preclinical findings demonstrating that low doses of bortezomib together with NPI-0052 trigger a potent anti-MM effect in vitro and in vivo, suggests the promise of combination treatment strategies to enhance anti-MM activity, reduce toxicity, overcome drug resistance, and improve outcome in MM patients. In addition to the above studies, data related to combination of NPI-0052 with lenalidomide (Revlimid™) and with histone deacetylase inhibitors such as MS-275, Tubacin or LBH589, will be presented.
Expression of V617F JAK2 in Mice Leads to MPD Mimicking Human ET, Idiopahtic Myelofibrosis, and PV.
Authors: Shide Kotaro , Shimoda Haruko K. , Kumano Takashi , Karube Kennosuke , Kameda Takuro , Oku Seido , Takenaka Katsuto , Suzuki Keiko , Kubuki Youko , Harada Mine , Shimoda Kazuya 
Published: April 2020

Abstract
An acquired JAK2 V617F mutation has been detected in up to 90% of patients with polycythemia vera (PV) and in a sizeable proportion of patients with other myeloproliferative disorders such as essential thrombocythemia (ET) and idiopathic myelofibrosis (IMF). However, how a single mutation may be responsible for such different clinical phenotypes is unknown. Mice transplanted with bone marrow cells in which V617F JAK2 was retrovirally expressed developed PV-like features, but not ET or IMF. To address the contribution of this mutation to the pathogenesis of these three MPDs, we generated transgenic mice expressing V617F JAK2 driven by the murine H2Kb promoter. We established two lines. The expression of V617F JAK2 mRNA in bone marrow cells was 0.45 and 1.35 that of endogenous wild-type JAK2 in the two lines. One line showed leukocytosis after 4 months of age, with a predominance of granulocytes. Among 43 mice, examined after 3 months of age, 8 (19%) showed polycythemia and 14 (33%) showed thrombocythemia. Two polycythemia cases also showed thromobocytosis. The other line showed extreme leukocytosis and thromobocytosis at one month of age. The leukocytosis progressed as the animals aged, but the thrombocytosis tended to resolve at 8 months. They showed anemia that means Hb value from 9 to 10 g/dL at one month old. Myeloid cells and megakaryocytes were predominant in the bone marrow of these animals, and splenomegaly with myeloid cell and megakaryocyte invasion was observed. We conclude that in vivo expression of V617F JAK2 results in ET-like, IMF-like, and PV-like disease.