bluuebunny/crossref_metadata_2025_split huggingface dataset:
Acetylation of an Ets Transcription Factor PU.1 Suppresses Its Transcriptional Activity.
Authors: Mouri Fumihiko , Tsukada Junichi , Fukamizu Akiyoshi , Tanaka Yoshiya 
Published: April 2020

Abstract
PU.1, a member of the Ets family transcription factors, is expressed restrictively in hematopoietic cells including monocytes and macrophages, and plays critical roles in the inflammatory responses and the development of hematopoietic cells. CREB-binding protein (CBP) regulates transcription by acetylating not only histones but also certain transcription factors. Here, we demonstrated that a specific inhibitor of histone deacetylases, trichostatin A (TSA) inhibits PU.1 transcriptional activity in monocytes and further showed that deletion of a histone acetyltransferase (HAT) domain of CBP resulted in synergistic cooperativity between CBP and PU.1. When human monocytic cells THP-1 were treated with TSA, our immunoprecipitaion and western blot assay showed that TSA enhanced PU.1 acetylation. Next, we investigated the effect of TSA on the transcriptional regulation of PU.1-dependent gene promoters such as the human prointerleukin 1β (IL1B) gene and the human granulocyte-macrophage colony-stimulating factor receptor α (GM-CSFRα) gene in transient transfection studies. Two distinct luciferase reporter plasmids (Luc) for the IL1B gene promoter and the GM-CSFRα gene promoter, IL1B-Luc and GM-CSFRα-Luc were used. When these plasmids were transiently transfected into THP-1 cells, TSA suppressed LPS-induced activities for the IL1B promoter and the GM-CSFRα promoter in a dose-dependent manner. In contrast, when NF-κB luciferase reporter, NF-κB-Luc was transfected into THP-1 cells, TSA synergistically increased LPS-induced NF-κB activities. Moreover, when a PU.1 expression vector, pECEPU.1 was cotransfected into PU.1-deficient murine thymocytes EL4 along with either IL1B-Luc or GM-CSFRα-Luc. The PU.1-induced promoter activities were strongly suppressed through TSA treatment. FACS analysis further indicated that TSA suppressed LPS-induced expression of IL-1β and GM-CSFRα proteins. In addition, our EMSA data showed that TSA treatment did not affect DNA binding activity of PU.1 to the IL1B promoter. PU.1 has been shown to interact physically with CBP to transactivate their target genes. In our study, expression vectors for CBP wild-type or with a deletion of its HAT domain was cotransfected into EL4 cells along with IL1B-Luc and pECEPU.1. The HAT activity-deficient mutant showed synergistic transcriptional activity with PU.1 more strongly than the wild-type CBP. In this regard, our GST-pulldown assay showed that deletion of CBP HAT domain did not change binding affinity of CBP for PU.1. Our results propose a novel molecular mechanism by which PU.1-dependent genes is negatively regulated by HAT-induced acetylation in monocytes.
Toll-Like Receptor-Induced Expression of the NKG2D Ligand MICA on Monocytes Modulates Their Interaction with NK Cells.
Authors: Kloss Mercedes , Decker Patrice , Krusch Matthias , Baltz Katrin M. , Baessler Tina , Kanz Lothar , Steinle Alexander , Salih Helmut R. 
Published: April 2020

Abstract
Reciprocal interactions of NK cells with Dendritic Cells (DC) have been shown to influence activation of NK cells as well as maturation or lysis of DC and subsequent adaptive immune responses. However, little is known about the interaction of peripheral blood Monocytes with NK cells. Here we report that stimulation of Toll-like receptors (TLR) on Monocytes using LPS, Pam3Cys, R848 and PolyI:C induced mRNA and surface expression of MICA but no relevant changes of the further ligands of the activating immunoreceptor NKG2D, MICB or ULBP1-3. MICA upregulation upon stimulation with TLR ligands correlated with CD80 and HLA-DR upregulation and increased secretion of the pro-inflammatory cytokines IL-6, IL-8 and TNF by Monocytes. Interestingly, high MICA surface expression on activated Monocytes was not accompanied by a release of MICA in soluble form. The LPS-induced MICA upregulation by Monocytes was highly sensitive since it was observed with as little as 4 ng/ml LPS after 15 hours and was associated with IL-6 secretion and inhibition of apoptosis indicating that Monocytes were activated and functional. To determine whether upregulated MICA expression on Monocytes was detected by NK cells we took advantage of the fact that NKG2D is down-regulated after interacting with its ligands. While NKG2D levels on NK cells were not substantially changed in cultures with unstimulated Monocytes, a marked reduction of NKG2D was observed in the presence of activated MICA-expressing Monocytes. The modulation of NKG2D did not occur when NK cells and monocytes were separated by a transwell filter, which demonstrates that the numerous cytokines produced by TLR-activated Monocytes are not responsible for the NKG2D modulation and confirmes the role of MICA in Monocyte-NK cell interaction. Importantly, TLR-induced MICA expression on Monocytes stimulated IFN-γ production of NK cells, which could be reduced by addition of blocking anti-MICA/B F(ab’)2 fragments while isotype control had no effect. This demonstrates that the observed stimulation of NK cells was in fact specifically due to NKG2D-NKG2DL interaction. Our data indicate that NKG2D-MICA interaction provides a mechanism by which NK cells and Monocytes may communicate directly during innate immune responses to infections in humans.
Dendritic Cells as a Novel Target for Erythropoietin: Studies in Murine Models.
Authors: Lifchitz Lilach , Sagiv Sari Prutchi , Markovitz Maayan , Mittelman Moshe , Newmann Drorit 
Published: April 2020

Abstract
Erythropoietin (EPO) is the major hormone that promotes the proliferation and differentiation of erythroid progenitor cells. Unexpectedly, EPO receptor (EPO-R) was also found on non-erythroid cells; thus leading to the discovery of non-erythroid effects of EPO. Our own previous contribution to that issue was in demonstrating that the immune system is a target for EPO, including both the cellular and humoral immune response types. As yet, the direct target cells for EPO as well as the molecular mechanisms underlying its function as an immunomodulator remain unknown. We first examined lymphocytes as possible candidates, and could not detect any expression of EPO-Rs on these cells. Here, we focused on dendritic cells (DCs), known to initiate immune response as antigen presenting and T cell priming cells. We employed murine bone marrow DCs (BMDCs) and splenic DCs (SDCs) models to determine EPO-R expression, and delineate in-vitro and in-vivo effects of EPO via these cells. We found that BMDCs express EPO-R mRNA, as detected by RT-PCR. In vitro stimulation of the BMDCs with recombinant human EPO (rHuEPO) activated the NFkB and MAPK signaling pathways, and induced a higher surface expression of CD80, CD86 and MHC class II. These data are reinforced by in vivo experiments, showing that rHuEPO injection into naïve mice led to an increase in the SDC population and in the cell surface expression of CD80, CD86 and MHC class II markers. These novel findings implicate the significance of the multifunctional role of EPO in the hematopoietic and immune systems, and may lead to its further clinical applications as an immunomodulator.
Role of Novel C-Type Lectin Receptor DCL-1 in Phagocytes and Dendritic Cells.
Authors: Hart Derek N.J. , Khan Seema , d’Aniello Elisabetta , McDonald Kylie J. , Kato Masato 
Published: April 2020

Abstract
Cell surface C-type lectins are implicated in many aspects of immunity. For example, professional phagocytes utilise cell surface C-type lectins in pathogen recognition/binding and phagocytosing the pathogens for destruction and/or antigen processing (e.g. mannose receptor, DC-SIGN, dectin-1). Some C-type lectins are involved in cell-cell adhesion and trafficking of leukocytes (e.g. selectins). DCL-1 is a small type I transmembrane C-type lectin receptor, discovered as a genetic fusion partner of DEC-205. DCL-1 protein was highly conserved amongst the human, mouse and rat orthologs. The human DCL-1 gene, composed of 6 exons, was located to a cluster of type I transmembrane C-type lectin genes on chromosomal band 2q24. Multiple expression array, RT-PCR and FACS analysis using new anti-hDCL-1 mAbs established that DCL-1 expression in leukocytes was restricted to monocytes, macrophages, granulocytes and DC, although DCL-1 mRNA was present in many tissues. Stable hDCL-1 CHO cell transfectants endocytosed FITC-conjugated anti-hDCL-1 mAb rapidly (t1/2 = 20 min) and phagocytosed anti-hDCL-1 mAb-coated microbeads, indicating that DCL-1 may act as an antigen uptake receptor. However, anti-DCL-1 mAb-coated microbead binding and subsequent phagocytic uptake by macrophages was ∼8-fold less efficient than that of anti-macrophage mannose receptor (MMR/CD206) or anti-DEC-205/CD205 mAb-coated microbeads. Confocal studies showed that DCL-1 colocalized with F-actin in filopodia, lamellipodia and podosomes in macrophages and the colocalization was not affected with cytochalasin D, whereas the MMR/CD206 and DEC-205/CD205 did not colocalize with F-actin. Furthermore, DCL-1-EGFP transiently expressed in COS-1 cells colocalized with F-actin at cellular cortex and microvilli. These data suggest that hDCL-1 is an unconventional lectin receptor that plays roles not only in endocytosis/phagocytosis, but also in myeloid cell adhesion and migration.
Hematopoietic miRNAs Expression in Myelodysplastic Syndromes.
Authors: Pons Aina , Nomdedeu Benet , Navarro Alfons , Gaya Anna , Pairet Silvia , Granell Miquel , Valera Sandra , Gel Bernat , Montserrat Emili , Monzo Mariano 
Published: April 2020

Abstract
Myelodysplastic syndromes (MDS) are clonal hematopoietic stem-cell disorders that include a broad spectrum of entities characterized by ineffective hematopoiesis and risk of transformation into acute myeloid leukemia (AML). MicroRNAs (miRNA) act as negative expression regulators of important genes as those participating in cellular proliferation, apoptosis and carcinogenesis. In this study we analyzed the expression patterns of 26 miRNA hematopoietic-related genes in patients with MDS. Total RNA was extracted from 20 MDS patients (BM and PB) and 5 normal controls. Median age of patients was 70 (range, 55–82) years. Seven patients had refractory anemia with excess of blasts (RAEB-I, n=5; RAEB-II, n=2), 7 refractory cytopenia with multilineage dysplasia (RCMD), 3 RCMD with ringed sideroblasts (RCMD-RS), 1 chronic myelomonocytic leukemia (CMML), 1 refractory anemia with ring sideroblasts (RARS) and 1 MDS associated with isolated del(5q). 15 patients presented low risk IPSS (LR) (low or intermediate-1) and 5 patients had high risk IPSS (HR) (intermediate-2 and high risk). Twenty-six mature miRNAs were assessed by Stem-loop RT-PCR and Real time PCR in ABI PRISM 7500. MiRNA expression data was normalized to overall median and relative quantification was calculated with the 2−ΔΔCt method. The data were presented as log10 of relative quantity of target miRNA. Median of normal controls was used as calibrator for all samples. Data were analyzed by class comparison methods using BRB Array Tools version 3.4.0 and TIGR multiexperiment viewer. Hierarchical clustering analysis of BM categorized two clusters corresponding to BM-MDS and BM-controls. Then, miR-142-5p (p&lt;0.001) and miR-18a (p&lt;0.001) were differentially expressed between BM-MDS and BM-controls while miR-103 (p=0.003) was overexpressed and miR-18a (p=0.02) was underexpressed in PB-controls with respect to PB-MDS. The analysis of miR-10a and miR-103 in PB (p=0.035 and p=0.031) and miR-181a and miR-10a in BM (p=0.044 and p=0.02) allowed us to discriminate between LR and HR groups. We also found that miR-222 was overexpressed in AML regarding MDS samples (p&lt;0.001). It has been previously described that high activity of miR-222 is necessary to maintain low p27KIP1 levels and continuous proliferation in certain cancer cell lines. In conclusion, this study confirms that miRNAs have a role in the pathogenesis of MDS and their transformation into AML.
High-Resolution Analyses of Epigenetic Aberrations in Myelodysplastic Syndrome.
Authors: Yamamoto Go , Nakamura Fumihiko , Iio Mitsuru , Kato Motohiro , Nannya Yasuhito , Sanada14 Masashi , Hangaishi Akira , Chiba Shigeru , Mori Hiraku , Kurokawa Mineo , Watanabe Toshiki , Omine Mitsuhiro , Ogawa Seishi 
Published: April 2020

Abstract
Myelodysplastic syndromes are heterogeneous groups of clonal hematopoietic disorders characterized by ineffective blood cells production and predisposition to acute myeloid leukemia, and as such, it is well established that these syndromes actually represent neoplastic processes in which a series of gene mutations accumulate in blood cell precursors, leading to neoplastic expansion of dominant clones. During the past two decades, a number of genetic abnormalities have been described in MDS cases, including copy number alterations of particular chromosomal segments, mutations of Ras, p53, runx1, and Flt3 genes, and translocation involving Evi-1 family genes, TEL, MLL and Nup98 genes. On the other hand, epigenetic abnormalities are also thought to play an important role in the pathogenesis of MDS, because demethylating agents such as 5-azacydine and decitabine are often effective for high risk MDS. Unfortunately, however, only a few genes, such as INK4B gene, have been implicated in MDS pathogenesis. Especially, no genome-wide analysis of epigenetic changes in MDS has been reported. So, in the current study, we comprehensively investigated abnormalities of DNA methylation in 30 MDS specimens, using Affymetrix tiling array combined with methylated DNA immunoprecipitation (MeDIP). In this method, genomic DNA from MDS specimens was first fragmentized with ultrasonication and immuno-precipitated with anti-methylcytosine antibody (MeDIP). The immunoprecipitated DNA was then amplified by PCR and subjected to hybridization to the promorter tiling array. In this array, regulatory regions of more than 25,000 genes are tiled by 6.5 millions of oligonucleotide probes to enable sensitive detection of target sequences and approximately 59% of CpG islands in the human genomes are covered in a single array. The extent and distribution of methylation were highly variable between specimens, although some CpG islands, such as p15INK4B and HOX gene clusters, seemed to be commonly involved in different cases (Figure). In conclusion, MeDIP on chip analysis could be a powerful method for genome-wide detection of DNA methylation and facilitate our understanding of the pathogenesis of MDS.
Figure Figure
High-Resolution CGH Analysis of CD34+ Cells from Lower-Risk Myelodysplastic Patients Reveals Cryptic Copy Number Alterations and Predicts Overall and Leukemia-Free Survival.
Authors: Starczynowski Daniel T. , Vercauteren Suzanne , Telenius Adele , Sung Sandy , Tohyama Kaoru , Brooks-Wilson Angela , Spinelli John J. , Eaves Connie J. , Eaves Allen C. , Horsman Douglas E. , Lam Wan L. , Karsan Aly 
Published: April 2020

Abstract
The myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematologic malignancies characterized by peripheral cytopenias, a hypercellular marrow with ineffective hematopoiesis and a propensity to progress to acute myeloid leukemia (AML). MDS is thought to arise from a primitive (CD34+) hematopoietic cell that has acquired genetic and/or epigenetic abnormalities. Risk stratification according to the International Prognostic Scoring System (IPSS) defines patient categories that correlate with survival and the likelihood of transformation to AML. Greater than 50% of individuals diagnosed with MDS are in the lower-risk groups. The importance of cytogenetics in risk stratification has been verified in several studies, but is of limited value in patients with lower-risk subtypes because approximately 50% of these patients do not have karyotypic abnormalities detectable using standard techniques. To further our biological understanding of lower-risk subtypes of MDS, and to identify potential MDS-initiating alterations in the genome, we looked for alterations in DNA extracts from purified CD34+ marrow cells from 44 MDS lower-risk patients using a submegabase bacterial artificial chromosome (BAC) array to perform whole genome comparative genomic hybridization (CGH) analyses. These studies identified numerous cryptic structural DNA alterations that were not detectable by standard cytogenetic analysis and were also not found in 15 age-matched normal controls. Although most patients tested had a normal karyotype, 23 recurring, novel copy number alterations of a median size of 0.6 megabases were identified. These included gains at 11q24.2-qter, 17q11.2 and 17q12, and losses at 2q33.1-q33.2 and 14q12. Comparison of changes in CD34+ marrow cells with DNA from CD3+ cells isolated from the same patients showed that a recurring duplication at band 17q12 was exclusive to the CD34+ cells in 2 of 3 patients. Validation of this copy number gain at chr 17q12 by FISH confirmed duplication of the locus. In addition, whole genome array CGH analysis of CD34+ marrow cells from karyotypically normal (n = 25) and abnormal (n = 15) lower risk MDS patients revealed extensive genome alterations (involving &gt;3 Mb) correlated with poorer overall survival, and was more frequently associated with transformation to AML as compared to IPSS stratification alone. Our studies suggest that array CGH may be useful as an ancillary test to better stratify lower-risk subtypes of MDS and, at the same time lead to the identification of early mutations that contribute to disease initiation.
Dependency of Survival on Additional Karyotype Anomalies and Bone Marrow Blast Count in Patients Suffering from Myelodysplastic Syndromes with 5q Deletion.
Authors: Knipp Sabine , Hildebrandt Barbara , Jotterand Martine , Schapira Marc , Giagounidis Aristoteles , Gattermann Norbert , Aul Carlo , Germing Ulrich 
Published: April 2020

Abstract
The 5q deletion syndrome is described as having a favourable outcome in MDS. However, little data is available concerning del(5q) and additional karyotype anomalies or taking into account bone marrow blast count. We screened the MDS registries of Düsseldorf, Germany and Lausanne, Switzerland in order to obtain data on patients with karyotype anomalies including del(5q). In both registries 1073 patients were karyotyped at diagnosis. 198 patients with del(5q) with or without other karyotype anomalies and 105 patients with complex karyotype anomalies without del(5q) were analysed. 106 patients showed del(5q) as single anomaly, 23 had 1 additional karyotype anomaly, 69 had 2 or more additional anomalies and were therefore classified as complex karyotype. In the group with del(5q) only, mean survival was 76 months as compared to 42 months in patients with a normal karyotype. In the group with 1 additional anomaly mean survival was 47 months (p=0.02), in the group with 2 or more additional anomalies 7 months (p=0.0005). Survival differed significantly in each subgroup when patients presenting less than 10% of bone marrow blasts where compared to patients with 10% or more. Patients with del(5q) only show a median survival of 12 months when presenting with more than 10% medullary blasts. Patients with a complex karyotype including del(5q) with less than 5% medullary blasts had a median survival of 12 months, but median survival was only 6 months in patients with elevated medullary blasts. By multivariate analyses, we found that additional karyotype anomalies, followed by an elevated medullary blast count above 10% are the most important independent prognostic parameters. Furthermore, we compared the subgroups with del(5q) and 2 or more additional karyotype anomalies (n=69) to patients with complex karyotype anomalies not including del(5q) (n=105). Median survival of the group with complex karyotype anomalies including del(5q) was 7 months as compared to 12 months in the group without del(5q) (p=0.02). 12 months after diagnosis, 30% of the group with del(5q) in a complex karyotype was alive as compared to 45% of the group with a complex karyotype without del(5q). Disease related death was noted in 77% of patients with a complex karyotype and del(5q) and 73% without del(5q). Therefore, our data show that the prognosis of survival in patients with del(5q) is highly dependent on the number of additional karyotype anomalies as well as medullary blast count. The prognosis of MDS patients with del(5q) is associated with an extremely poor prognosis when diagnosed within a complex karyotype. Although there are no substantial differences in clinical, haematological and morphological data between patients with or without del(5q), the median survival differs significantly.
Figure Figure
Human Leukocyte Antigen DR15 in Patients with Myelodysplastic Syndromes (MDS) – A Study in Chinese Patients.
Authors: Master Qiong Liao , Zhang Yan , Li Xiao , Zhang Zheng , Ying Shaoxu , Master Chunkang Chang , Pu Quan ,   (Intr. by H. Joachim Deeg)
Published: April 2020

Abstract
We investigate the frequency of the human leukocyte antigen DR15 (HLA-DR15) allele in patients with myelodysplastic syndromes (MDS). We used polymerase chain reaction-sequence-specific primers (PCR-SSP) to detect HLA-DR15 in the peripheral blood of patients with MDS(n = 76) and healthy controls (n = 227).
The frequency of HLA-DR15 in MDS patients (40.8%) was significantly higher than in controls (13.7%; P &lt; 0.01). The diagnoses of refractory anemia/refractory anemia with ring sideroblasts (RA/RARS) accounted for 77.4% (24/31) and 62.2% (28/45) of the DR15-positive and the DR15-negative patients, respectively (difference not statistically significant). Although no statistically significant difference was observed, some trends were observed: IPSS low-risk MDS (IPSS score, ≤1) accounted for 80.6% of the DR15-positive patients compared to 64.4% among the DR15-negative patients. However, the difference between the numbers of DR15-positive and DR15-negative patients with chromosomal abnormalities was not statistically significant. Nevertheless, poor risk chromosome abnormalities (according to IPSS), were present in only 1 DR15-positive patient, while such abnormalities were present in 8 DR15-negative patients. In addition, the proportions of DR15-positive and DR15-negative patients with more than 5% blasts in marrow were 19.4% and 31.1%, respectively. Peripheral blood pancytopenia occurred in 51.6% of DR15-positive, and in 40.0% of DR15-negative patients. Although the HLA-DR15 allele appeared to be present more frequently in patients less than 60 years of age, this association was not significant. The frequency of HLA-DR15 was significantly higher in patients with MDS than in healthy controls suggesting the possibility that HLA-DR15 is associated with an enhanced susceptibility to develop MDS. The fact that HLA-DR15 was predominantly noted in patients with RA/RARS and low IPSS scores, suggested that HLA-DR15 might be associated more with bone marrow failure and less with leukemic transformation.
clinical/experimental characteristics in HLA-DR15 positive or negative MDS patients Cohort HLA-DR15 positive(n=31) HLA-DR15 negative(n=45) P value RA/RARS(case/%) 24/31(77.4) 28/45(62.2) 0.161 Low risk (IPSS≤1) (case/%) 25/31(80.6) 29/45(64.4) 0.126 Karyotype abnormal(case/%) 13/31(41.9) 18/45(40.0) 0.866 Poor chromosome(case/%) 1/31(3.2) 8/45(17.8) 0.117 Blast&gt;5%(case/%) 6/31(19.4) 14/45(31.1) 0.253 Pancytopenia(case/%) 16/31(51.6) 18/45(40.0) 0.317 Male patients(case/%) 17/31(54.8) 25/45(55.6) 0.951 age(&lt;60 years) (case/%) 20/31(64.5) 20/45(44.4) 0.085
Figure Figure Figure Figure