bluuebunny/crossref_metadata_2025_split huggingface dataset:
Roles of HLA-DR15 Alleles in the Pathogenesis of Acquired Aplastic Anemia (AA): DRB1*1502 Is in Linkages Disequilibrium with Unique Genes Which Determine Susceptibility to AA.
Authors: Sugimori Chiharu , Mochizuki Kanako , Feng Xingmin , Nakao Shinji 
Published: April 2020

Abstract
It is well known that HLA-DRB1 alleles corresponding to HLA-DR15 such as DRB1*1501 and DRB1*1502 are associated with susceptibility to acquired aplastic anemia (AA) and paroxysmal nocturnal hemoglobinuria (PNH). However, little is known about how these DRB1 alleles contribute to the development of AA. We previously reported that DRB1*1501 is significantly associated with both an increase in the proportion of PNH-type cells and good response to a immunosuppressive therapy (IST) including antithymocyte globulin (ATG) plus cyslosporine (CsA) or CsA alone, while DRB1*1502, whose DR molecules differ from those derived from DRB1*1501 in only one amino acid, is not associated with either of the two factors. To gain insight into the reason for different contributions to the development of AA by the two DRB1 alleles, we determined DRB1 alleles and the proportion of PNH-type cells of 77 patients with recently diagnosed AA and analyzed the relationship of DRB1 alleles and the presence of increased PNH-type cells with response to ATG plus CsA therapy. Fifty-five of 77 patients (71%) improved with ATG plus CsA therapy. When the factors favorably affecting the response to IST in the AA patients were examined under a multivariate analysis, only the presence of PNH-type cells was significantly associated with the response to IST. Kaplan-Meier curves showed that there were significant differences in the probability of the response to IST between the DRB1*1501 + 1502 − patients and either the DRB1*1501 − 1502 + patients (P &lt;0.01) or the DR15- patients (P =0.01). However, these differences in the probability of response to IST were no longer observed when the probability of response was compared in either the PNH + patients or the PNH − patients (Figure). Although DRB1*1502 was not associated with a good response to IST, the markedly high frequency of the allele in patients aged over 40 (52%) prompted us to study genes in linkage disequilibrium with DRB1*1502 which may determine susceptibility to AA. We compared the frequencies of 30 microsatellite marker alleles on chromosome 6 among three different groups of individuals possessing DRB1*1502 which include 18 PNH + AA patients, 13 PNH − AA patients, and 29 healthy individuals. There were no significant differences in the frequency of any of the polymorphic makers between PNH + AA patients and healthy individuals. On the other hand, one of the microsatellite maker alleles (C1-3-1) in close proximity to HLA-C locus was found at a significantly higher (Pc =0.01) frequency in PNH − AA patients than in PNH + AA patients and healthy individuals. These data suggest that DRB1*1501 contributes to development of AA via immune mechanisms leading to an increase in the proportion of PNH-type cells while DRB1*1502 is involved in the pathogenesis of AA by way of other genes in linkage disequilibrium with this DRB1 allele.
Figure Figure
Clinical Implications of Abnormal Cytogenetics at Diagnosis of Aplastic Anemia.
Authors: Kim Sung-Yong , Lee Jong-Wook , Cho Byung-Sik , Eom Ki-Seong , Kim Yoo-Jin , Lee Seok , Min Chang-Ki , Kim Hee-Je , Cho Seok-Goo , Kim Dong-Wook , Min Woo-Sung , Kim Chun-Choo 
Published: April 2020

Abstract
Because cytogenetic abnormalities of aplastic anemia at diagnosis have been reported fairly infrequently, their clinical implications have not known yet. A retrospective study was performed of the cytogenetics findings and clinical courses in patients with typical morphological and clinical features of aplastic anemia from a single institution for the years 1995 through 2005. The results of chromosome analysis of 610 patients were evaluable. Of the evaluable patients, 584 (95.7 %) had normal karyotypes and 26 (4.3 %) had abnormal karyotypes at diagnosis. The most frequent abnormality was trisomy 8 (n=13) followed by deletion 1q (n=5) and monosomy 7/deletion 7q (n=5). Other chromosome abnormalities were isochromosome 17q (n=1), trisomy 15 (n=1) and monosomy 21 (n=1). Among the 584 patients with typical aplastic anemia and no cytogenetic abnormalities, only two developed MDS/AML during the follow-up period, while 5 (19.2%) of 26 patients with typical aplastic anemia and abnormal cytogenetics subsequently developed MDS/AML. The incidence of secondary MDS/AML was statistically higher in abnormal cytogenetics group compared with normal cytogenetics group (p&lt;0.001). The incidence of secondary MDS/AML was not influenced by immunosuppressive therapy (IST) (p=0.715). The patients with trisomy 8 responded poorly to immunosuppressive therapy (IST) and showed statistically significant lower response rate compared with the patients with other cytogenetics (p=0.033). However, response rates of IST were not statistically different in the patients with normal cytogenetics group and the patients with abnormal cytogenetics other than trisomy 8 (p=1.000). Four patients with abnormal cytogenetics received allogeneic hematopoietic stem cell transplantations (allo-HSCT) with the same conditioning as the patients with normal cytogenetics. Three of them are still alive with normal peripheral blood counts. One of them died of acute GVHD and infection after successful engraftment. Our analysis suggested that cytogenetic abnormalities at diagnosis of aplastic anemia could be a risk factor for development of secondary MDS/AML and the patients with trisomy 8 at diagnosis of aplastic anemia might hardly respond to IST. Outcomes of allo-HSCT for aplastic anemia with abnormal cytogenetics probably are not different compared with aplastic anemia with normal cytogenetics.
Significance of Copy Number Alterations for Molecular Treatment Response in Childhood Acute Lymphoblastic Leukemia.
Authors: Steinemann Doris , Cario Gunnar , Stanulla Martin , Karawajew Leonid , Tauscher Marcel , Weigmann Anja , Göhring Gudrun , Ludwig Wolf-Dieter , Harbott Jochen , Radlwimmer Bernhard , Bartram Claus-Rainer , Lichter Peter , Schrappe Martin , Schlegelberger Brigitte 
Published: April 2020

Abstract
In vivo response to initial therapy, as assessed by determination of minimal residual disease after five and 12 weeks of treatment, has evolved as one of strong prognostic factors in children with acute lymphoblastic leukemia (ALL) treated according to the BFM regime. It is currently not known if the individual treatment response might be influenced by copy number alterations (CNA) leading to altered gene expression. We compared leukemic genomic profiles of 25 treatment sensitive (MRD-SR) and 25 resistant (MRD-HR) childhood ALL patients by means of high-resolution array-CGH. CNA were found in 46 patients (92%) of both treatment response groups. Microscopic alterations affecting the whole or nearly whole chromosome arm were frequently found, e.g. gain of 21 in 11/50, loss of 9p in 5/50, loss of 8p in 3/50, loss of 20q in 3/50 and loss of 7p in 2/50 or gain of 1q in 2/50. The most significant difference was a gain of chromosome 1q23-qter due to an unbalanced t(1;19), found in 10/25 MRD-SR patients, but in none of the MRD-HR patients (p&lt;0.002). The most frequent CNA in the MRD-HR group were deletions of genomic regions harboring the immunoglobulin genes (Ig), e.g. 2p11.2 in 15 of 25 cases (60%) compared to 7 of 25 in the MRD-SR group (28%) (p=0.045). Combining all Ig loci, significantly more MRD-HR than MRD-SR patients were affected with deletions (17 versus 8 patients, p=0.02). The frequency of other CNA, like loss of 9p21 or gains of 21q, did not differ strongly between the two patient groups. This is the first study evaluating the clinical significance of CNA as detected by array-CGH in childhood ALL and may lead to improved risk classification.
The Impact of Transfusion-Dependency on the Outcome of Patients with Myelodysplastic Syndromes (MDS) Receiving Allogeneic Peripheral Blood Stem Cell Transplantation after Myeloablative Conditioning.
Authors: Platzbecker Uwe , Germing Ulrich , Stumpf Julian , Kröger Nikolaus , Böhm Anne , Valent Peter , Schwerdtfeger Rainer , Krieger Otto , Kobbe Guido , Ehninger Gerhard , Bornhäuser Martin 
Published: April 2020

Abstract
Introduction: The majority of MDS patients become red blood cell transfusion-dependent during their clinical course. In fact, the presence or absence as well the extent of red blood cell transfusion-dependency (RBC-TD) has been recently shown to add significant prognostic information for an individual MDS patient treated with supportive care (BSC) only. Given the fact that allogeneic hematopoietic cell transplantation (HCT) is the only curative option for MDS patients, the aim of this study was to elucidate the role of RBC-TD on patient outcome.
Methods: We report results of a retrospective multicenter German-Austrian study investigating 100 patients with MDS (RA(RS) n=33, RAEB n=33, RAEB-t n=19, CMMOL n=11, MDS/AML n=4) with either IPPS LOW (n=2), INT-1 (n=31), INT-2 (n=37) or HIGH (n=30) undergoing allogeneic myeloablative conditioning followed by peripheral blood stem cells (PBSC) from related (n=39) or unrelated donors (n=61). The median age of the patients was 50 years (range 18–68).
Results: With a median follow-up of 32 months the 2-year overall survival (OS) was 48% for all patients and 64%, 50% and 34% for patients according to IPSS INT-1, INT-2 or HIGH, respectively (p=0.03 for INT-1 vs. HIGH). The 2-year OS of all patients was not different when comparing 68 patients displaying vs. 24 not displaying RBC-TD prior to PBSCT (46% vs. 55%, p=0.67). This holds also true when analyzing INT-1/INT-2 (n=62) as well as HIGH (n=27) patients separately. In a multivariate analysis only FAB classification (p=0.03) but not prior induction chemotherapy (n=18), presence of fibrosis (n=18), IPSS LOW/INT-1 vs. INT-2/HIGH, RBC-TD as well as ferritin level lower or above the median 973 μg/l (range 19–7000) had an impact on OS.
Conclusion: These data suggest that the negative prognostic impact of RBC-TD in MDS patients receiving BSC can be overcome by myeloablative allogeneic PBSCT.
Methylation Profiles of p16, p15 and p14 Genes in Korean Patients with Multiple Myeloma.
Authors: Kang Seong-Ho , Lee Dong Soon , Kim Tae Young , Min Hyun Jung , Oh Bo Ra , Cho Han-Ik , Yoon Sung-Soo ,   the Korean Multiple Myeloma Working Party
Published: April 2020

Abstract
Dysruption of cell cycle control genes (p16, p15 and p14) is known to be involved in the tumorigenesis of multiple myeloma (MM). We investigated the inactivation status of p16, p15 and p14 genes using promoter methylation study and fluorescent in situ hybridization (FISH) study in patients with MM and analyzed the association of inactivation of those genes and clinical prognosis.MM Promoter methylation study of p16, p14 and p15 gene was done with newly diagnosed 52 patients with by bisulfite modification and methylation specific PCR. Two sets of primers were used for p16 methylation study and one set of primer was used for p15, p14 methylation study. Deletion of p16, p14 and p15 gene was detected by dual color FISH (Vysis, Downers GroveIL, USA). Overall survival was analyzed by Kaplan Meier Method and Cox’s proportional hazard model. Methylation of p16, p15 and p14 promotor was detected in 36/52 (69.2%), 15/52(28.8%) and 7/52 (13.5%) patients with MM, respectively. Methylation of any of p16, p15 or p14 promotor was observed in 44/52 (84.6%) of MM patients Deletion of p16, p15 and p14 was detected in only one of the patients with MM. Of note, in the cases (15/52, 28.8%) showing that in cases with two positive methylation specific PCR of the p16 promotor with two sets of primers, significant lower overall survival rate (p&lt;0.036) were observed compared with only one positive methylation specific PCR. Heavy methylation of p16 protomor was a independent predictive variable for overall survival [Hazard Ratio, 3.74; 95% CI, 1.44–9.68, P = 0.007]. Other adverse prognostic factors by univariate analysis were old age (p=0.014), β2-microglobuline (p=0.026), high serum creatinine levels (≥2.0 mg/dL, p=0.014). More than 80% of MM patients showed the methylation of any of p16, p15 or p14 promotor, which suggest the potential applicability of hypomethlyating agents to MM. The promoter methylation of p16, p14 or p15 was a major contributor to the disruption of cell cycle regulation in MM, whereas both the deletion and/or the promoter methylation of p16, p14, and p15 contributed to the disruption of cell cycle regulation in ALL. In our study, methylation of two positive methylation specific PCR of the p16 promotor was an independent adverse prognostic factor in MM. We infer that quantitative methylation study for p16 promotor is helpful for the evaluation of prognosis.
Figure 1. Overall survival analysis of patients with according to p16 methylation amount (P = 0.036) Figure 1. Overall survival analysis of patients with according to p16 methylation amount (P = 0.036)
FIP1L1/PDGFRa Synergizes with SCF/c-Kit Signaling To Induce Systemic Mastocytosis in a Chronic Eosinophilic Leukemia Murine Model
Authors: Yamada Yoshiyuki , Cancelas Jose A. , Brandt Eric B. , Sanchez-Aguilera Abel , McBride Melissa , Al-Moamen Nabel J.H. , Finkelman Fred , Williams David A. , Rothenberg Marc E. 
Published: April 2020

Abstract
Systemic mastocytosis (SM) associated with chronic eosinophilic leukemia (CEL)/hypereosinophilic syndrome (HES) is a result of expression of the Fip1-like1 (FIP1L1)/platelet-derived growth factor receptor alpha (PDGFRa) (F/P) fusion gene. We have previously described a murine CEL/HES model (CEL-like mice) induced by F/P fusion gene transduction and T-cell overexpression of IL-5 (Yamada Y et al., Blood 2006). We have now validated a preclinical murine model of F/P-induced SM/CEL and analyzed the pathogenesis of SM in this model. F/P+ mast cells (MC, defined as EGFP+/c-kit+/FceRI+) were significantly increased in the small intestine, bone marrow (BM) and spleen of CEL-like mice compared to wild-type mice (Table). CEL-like mice also developed cutaneous MC infiltration. In addition, mMCP-1 serum levels, which correlate well with MC expansion and activation in vivo, were significantly higher in CEL-like mice than in wild-type mice (64,000 ± 23,800 and 38 ± 41.4 pg/ml, respectively). F/P induces increased expansion of BM-derived MC in vitro (∼2,000-fold) and F/P+ BM-derived MC survive longer than wild-type MC in cytokine-deprived medium (28.0 ± 2.3% vs. 8.7 ± 3.1% 7AAD−/Annexin V− cells after 48 hours). This correlated with increased Akt phosphorylation in the F/P+ MC. Since c-kit mutations are the most frequent cause of SM, we analyzed the possible synergistic role of SCF and F/P signaling. F/P and SCF/c-kit signaling indeed synergize in the development of BM-derived MC (16-fold greater expansion than in the absence of SCF) and F/P+ BM-derived MC showed a 3.7-fold greater migratory response to SCF than wild-type BM-derived MC. In order to determine the role of SCF/c-kit signaling in F/P+ MC development, activation and tissue infiltration in vivo,these responses were evaluated in mice that were treated with a blocking anti-c-kit blocking antibody, ACK-2, or an isotype-matched control antibody. ACK-2 treatment suppressed intestinal MC infiltration and elevated plasma levels of mMCP-1 induced by F/P expression by 95 ± 6.0% and 98 ± 0.76%, respectively, whereas MC and plasma mMCP-1 were completely undetectable in wild-type mice treated with ACK2. This suggests that SCF/c-kit interactions may synergize with F/P to induce SM. In summary, mice with CEL-like disease also develop SM. F/P-induced SM is a result of increased in vivo MC proliferation, survival, activation and tissue infiltration. SCF/c-kit signaling synergizes with F/P in vivo and in vitro to promote mast cell development, activation and survival.
EGFP+/c-kit+/FcεRI+ cell frequency in tissues of control and CEL-like mice (%) Control mice CEL-like mice Small intestine 1.0±0.95 47±21.4* Bone marrow 0.2±0.14 3±1.9* Spleen 0.05±0.01 3±0.8*
Anti-Tumor Effect of INNO-406, a Novel Bcr-Abl Inhibitor in Combination with Cyclosporin A Against the Murine Central Nervous System and Systemic Leukemia Model.
Authors: Yokota Asumi , Kimura Shinya , Masuda Satohiro , Ashihara Eishi , Urasaki Yoshimasa , Terui Yasuhito , Ruthardt Martin , Ueda Takanori , Hatake Kiyohiko , Inui Ken-ichi , Maekawa Taira 
Published: April 2020

Abstract
Central nervous system (CNS) is one of the major cites for extramedullary relapse of Ph+ leukemias, which have been treated with imatinib mesylate (IM). The reason for this is that IM is a substrate for P-glycoprotein (P-gp) at the blood brain barrier and effluxed by it. We have already shown in the last annual meeting that INNO-406 had much stronger anti-tumor effects against the murine CNS leukemia model compared with IM, and INNO-406 is also effluxed from the murine CNS by P-gp. In this study, we investigated the combination effect of INNO-406 and P-gp inhibitors, verapamil or cyclosporin-A (CsA). First, we examined the growth-suppressive effect of INNO-406 and the combination with the P-gp inhibitors against the BCR-ABL positive leukemic cell line, K562 and the P-gp-overexpressing K562, K562/D1-9 cell line. K562/D1-9 showed 10 times higher resistant to both IM and INNO-406 compared with K562. Furthermore, both verapamil and CsA synergistically augmented the effect of INNO-406. Next, we investigated the pharmacokinetics of INNO-406 when orally administrated with CsA to mice. Mice were administrated p.o. with 50mg/kg of CsA 2 hours before INNO-406. We found that the concentration of INNO-406 in the CNS elevated by 2 times when combined with CsA, while the plasma concentration was decreased to two thirds of that when singly administrated with INNO-406. It was suggested that the decreased plasma concentration of INNO-406 seen here resulted from the increased uptake into the CNS by CsA inhibiting P-gp at the blood brain barrier. These changes of the drug distribution to the murine tissues may alter the anti-leukemic effect of INNO-406, thus we planned to investigate the combination effect of INNO-406 and CsA in the murine models of both CNS and systemic leukemia. We found that CsA significantly augmented the anti-tumor effect of INNO-406 in the CNS leukemia model. Moreover, in spite of the decreased plasma concentration of INNO-406, the combination with CsA also prolonged the survival phase of the mice in the systemic leukemia model, more significantly than single treatment of INNO-406 (Figure). This may be explained by which CsA increased the intracellular uptake of INNO-406, resulted from the direct inhibition of drug efflux via P-gp expressed in the leukemic cells. Phase I clinical study on INNO-406 is now underway in MD Anderson Cancer Center and in Frankfurt University. From the results of this study, we expected the effective application of INNO-406 in combination with P-gp inhibitor to the patients suffering from refractory Ph+ leukemia as well as CNS relapse.
Figure Figure
2-Phenylacetylenesulfonamide Upregulates Noxa and Induces p53-Independent Apoptosis of CLL Cells.
Authors: Steele Andrew J. , Prentice Archie , Cwynarski Kate , Hoffbrand A. Victor , Wickremasinghe R. Gitendra 
Published: April 2020

Abstract
The classical pathway of p53-mediated apoptosis involves transcriptional upregulation of pro-apoptotic proteins including Puma and Noxa. However, recent studies have identified a novel non-transcriptional mechanism mediated by direct binding of p53 to Bcl-2 or Bcl-XL at the mitochondrial surface and consequent neutralization of their anti-apoptotic function (Chipuk and Green, Cell Cycle, 2004; 3: 429–431). Induction of apoptosis of chronic lymphocytic leukaemia (CLL) cells by cytotoxic agents is strongly dependent on a functional p53 system and genetic changes which compromise p53 function result in drug resistance. We are therefore attempting to identify novel therapeutic agents which induce selective apoptosis of CLL cells by mechanisms independent of p53. Here we present data suggesting that 2-phenylacetylenesulfonamide (PAS; trivial name pifithrin μ) is a potential therapeutic agent for CLL. PAS was toxic to CLL cells, with a median LC50 of 10.72 μM (SD 3.8 μM n=20) as measured by a dye reduction cytotoxicity assay. Killing of CLL cells was by apoptosis, documented by the ability of 10–20 μM PAS to induce cytochrome c release from mitochondria (Fn2) to the cytosol (Fn1; Figure 1, upper), conformational change of Bax to a pro-apoptotic conformation, translocation of Bax from the cytosol to mitochondria (Figure 1, lower) and cleavage of the caspase 3 substrate poly(ADP ribose) polymerase (PARP). PAS toxicity was p53-independent, since p53-deleted CLL isolates were susceptible to killing. Furthermore, PAS was toxic towards leukemic cell lines with either inducible (Daudi), non-functional (Raji) or deleted (K562) p53. PAS treatment of CLL cells resulted in upregulation of Noxa and the subsequent displacement by Noxa of the pro-apoptotic Bim long (L) and extra-long (EL) isoforms from Bcl-2, as determined by western blot analysis of Bcl-2 immunoprecipitates (Figure 2). These observations contrast with those of Strom et al (Nat Chem Biol, 2006; 2: 474–479), who reported that PAS caused the displacement of p53 from mitochondria to the cytosol and thereby protected the cells from p53-dependent apoptosis. These differences may result from ectopic viral expression of p53 in the latter study whereas this protein was under normal cellular regulation in CLL cells. Normal T lymphocytes were relatively resistant to PAS, with IC50 values four times greater than those observed for CLL cells. In conclusion, our data suggest that PAS represents a novel class of agent which is toxic towards CLL cells via a mechanism involving p53-independent upregulation of Noxa and the subsequent unleashing of pro-apoptotic Bim from anti-apoptotic Bcl-2. This p53-independent toxicity may be of particular value in the treatment of drug-resistant CLL patients with compromised p53 pathways.
Figure 1. Figure 1. Figure 2. Figure 2.