bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mesenchymal Stem Cell Differentiation Markers Shared with Soft Tissue Sarcomas
Authors: Wirths Stefan , Buehring Hans-Joerg , Kanz Lothar , Hartmann Joerg T , Kopp Hans-Georg 
Published: April 2020

Abstract
Malignant tumors are hypothesized to harbor small populations of self-renewing cancer stem cells. Targeting these cells may be the decisive step to overcome treatment resistance and achieve tumor eradication in cancer patients. Advanced soft tissue sarcomas (STS) are rare tumors with a dismal prognosis and a small number of systemic treatment options. STS may originate from mesenchymal stem cells (MSC); the latter have mainly been isolated from adult bone marrow (BM) as non-hematopoietic, self-renewing cells whose in vitro progeny comprises osteoblasts, chondroblasts, myocytes, and adipocytes. While in vitro expression profiles of MSC have been investigated extensively, the in vivo counterparts of MSC are still hypothetical. To target rare human cell BM populations including MSC, an exclusive antibody panel was developed. The target antigens include platelet-derived growth factor receptor-β (CD140b), HER-2/erbB2 (CD340), the recently described W8B2 antigen as well as several surface antigens identified by novel antibodies. To define the expression pattern of MSC-markers in STS, three STS cell lines were tested for expression of these antigens. In addition, snap-frozen primary STS sections were analyzed by immunohistochemistry using the same antibody panel. All cell lines revealed expression of selected markers including CD340, W8B2, and CD140b. Several MSC markers were restricted to a subpopulation of cells. In addition, leiomyosarcoma cells displayed a different expression pattern as compared to liposarcoma and Ewing’s sarcoma cells. Results of immunohistochemistry analysis of primary leiomyosarcoma tumor samples correlated strongly with expression patterns established by FACS analysis. However, important cytoarchitectural features regarding selected markers were revealed by immunohistochemistry: while primary leiomyosarcomas displayed uniform expression of W7C6, HEK3D6, CD10, and CD318, other markers such as CD34, W5C5, and 57D2 were expressed by tumor endothelia only. Moreover, a population of perivascular tumor cells was found to express the MSC-markers W4A5, W8B2, CD140b, W3D5, and W5C4. Novel MSC-markers are expressed by subpopulations in STS cell lines as well as in primary sarcoma tissue. Further studies on the functional significance of these phenotypical studies are underway and may help to identify novel specific targets recognizing the self-renewing STS-compartment.
Gata1 Regulates Erythroid Transcription by Cooperating with Chromatin Remodeling Protein Snf2h
Authors: Podskocova Jarmila , Burda Pavel , Vargova Karin , Kokavec Juraj , Curik Nikola , Zavadil Jiri , Skoultchi Arthur , Stopka Tomas 
Published: April 2020

Abstract
Gata1 is transcription factor that regulates erythropoiesis and its direct interaction with chromatin remodeling protein Snf2h may affect chromatin structure (Rodriguez 2005). Snf2h belongs to SWI/SNF2 superfamily of ATPases regulating structure of nuclear chromatin by nucleosome movement and assembly. Snf2h knockout in mice is embryonic lethal and heterozygotes display mild growth retardation (Stopka 2003). We studied nuclear localization of Snf2h and detected its presence in euchromatin and to a lesser extent in heterochromatin. Decreased Snf2h levels in Snf2h heterozygotes and Snf2h-null embryos exhibit significantly decreased heterochromatin size. In addition, histone modifications associated with transcription activation (histone H3K79 dimethylation and H4K16 acetylation) are globally decreased in Snf2h mutants. To test the involvement of Snf2h in hematopoiesis, ectopically expressed Snf2h mutants were tested in Gata1-mediated transcription assay in HeLa cells and demonstrated that Snf2h efficiently repressed Gata1 transactivation. Testing whether the ATPase domain is required for the repression mechanism we found the Snf2h dominant negative mutant (DN) can also repress Gata1-dependent transcription in both HeLa and Snf2h +/− fibroblasts. We next studied the effect of Snf2h DN mutant on histone modifications downstream the Gata1 binding site and found that Snf2h DN further increases H3K79 dimethylation induced by Gata1. In contrast, an occupancy of histone H3 downstream the Gata1 binding site was significantly reduced by Snf2h DN mutant indicated it caused a defect in chromatin remodeling. Collectively, our data demonstrate a cooperative role of Gata1 and Snf2h in erythroid transcription regulation and propose that Snf2h in both ATP-dependent and ATPindependent manner represses transcription by disrupting the regular array of nucleosomes near Gata1 binding sites.
Identification of a Novel Acetyl-CoA Binding Protein (EST1) Which Epigenetically Regulates the Expression of Erythroid-Specific Genes
Authors: Fujiwara Tohru , Ikura Tsuyoshi , Ichikawa Satoshi , Takahashi Shinichiro , Furuyama Kazumichi , Igarashi Kazuhiko , Harigae Hideo 
Published: April 2020

Abstract
(Introduction) Embryonic stem (ES) cells that lack 5-aminolevulinate synthase-2 (ALAS2) gene provide a valuable model in dissecting molecular events in which heme is required during erythroid differentiation. Recently, we identified a novel acetyltransferase-like gene (EST1) through differential expression analysis between wild-type and heme-deficient erythroblasts as a potential downstream target gene of heme (BBRC2006;340:105–110, ASH 2007). EST1 belongs to the GNAT (GCN5-related N-acetyltransferase) superfamily since it contains the highly conserved amino acid residues (motif A), known as acetyl-CoA binding domain. Since the GNAT superfamily contains a wide variety of acetyltransferases with different substrates or with unknown functions, probing how EST1 might be required for biological events in erythroid cells, could be informative. Here, we investigated the role of EST1 during erythroid differentiation.
(Methods) EST1 was constitutively expressed using Flag/HA-tagged retroviral vector into mouse erythroleukemia (MEL) cell line. EST1 protein complex was purified by affinity chromatography from nuclear extract of EST1-expessed MEL cells. To obtain dominant-negative EST1-expressing cells, both Arg-62 and Gly-65 within EST1 were substituted to glutamic acid, and similarly transduced into MEL cells. These mutations have been widely applied for abolishment of acetyl-CoA binding activity. For depletion of endogenous EST1, siRNAs specific for EST1 were introduced into Hepa1c1c7 cells
(Results) Although recombinant EST1 protein did not have acetylase activity for free histones in vitro (ASH 2007), EST1 protein forms a multimeric protein complex. Western blot analysis using FLAG-eluted polypeptides revealed the presence of GCN5, TRRAP, SPT3 and GATA-1, implying that this protein complex might participate in the transcriptional regulation of erythroid-specific genes. Following EST1 depletion in Hepa1c1c cells, a significant decrease in the acetylation of H3 and a mild decrease in that of H4, were observed by Western blot analyses. Similarly, a significant decrease in the acetylation of H3 and a mild decrease in that of H4 were also observed in dominant-negative than in wild-type EST1-expressing MEL cells. Furthermore, the level of bmajor and ALAS2 mRNA were significantly lower in dominant-negative than in wild-type EST1-expressing MEL cells upon treatment with 1.5% DMSO for both 48h and 72h. We are currently exploring mechanisms of how EST1 participates in the regulation of histone modification in erythroid cells.
(Conclusion) EST1 may epigenetically regulate a subset of erythroid-specific genes under control of heme. Further elucidation of the function of the EST1 gene would enhance our understanding of the transcriptional network involving erythroid differentiation.
A Probabilistic Integrated Gene Network Model of Multiple Hematopoietic Lineage Specification
Authors: Fang Hai , Wang Kankan , Zhang Ji 
Published: April 2020

Abstract
Hematopoietic stem cells and their progenitor hierarchy are highly controlled by the underlying gene regulatory network. In past decades, great progress has been made in elucidating lineage-restricted transcription factors and lineage-specific gene expression patterns. With accumulation of genome-wide biological data, it is of great value to expand beyond mere transcriptional regulatory analysis to the systemic understanding. Here, we utilized a probabilistic integrated gene network for integrating heterogeneous functional genomic and proteomic data sources into predictive model of hematopoietic lineage diversification. We first constructed a naïve Bayesian network by incorporating disparate biological data, including time-series gene expression during re-dedifferentiation of leukemia along alternative paths into granulocyte or monocyte upon the treatment of differentiation-inducing agents, computationally generated transcription factor regulatory sites and microRNA targets, well-curated physical protein-protein interaction, and functional annotation data. The resultant network, coupled with binomial-based statistical analysis of the interplay between node properties and the network topology, predicted the specific hematopoietic lineage, generating testable hypotheses regarding their unified reprogramming principles. This study demonstrates the utility of the growing biological data in detailed elucidations of the orchestration of distinct hematopoietic cell fates.
Hematopoietic Stem/Progenitor Cell Mobilization in Donors and Recipients after Living Donor Liver Transplantation
Authors: Masuya Masahiro , Shiraki Katsuya , Ohishi Kohshi , Nakamura Shiho , Miyazaki Hiroshi , Katayama Naoyuki 
Published: April 2020

Abstract
Combination therapy with interferon-α and anti-viral drug is the current treatment of choice for chronic hepatitis viral infection. However, an important number of patients fail to respond to this therapeutic strategy and develop cirrhosis or liver cancer. Liver transplantation (mainly living donor liver transplantation (LDLT) in Japan) is known to be the ultimate therapy for them. To clarify the association of liver regeneration and hematopoietic stem/progenitor cells (HSCs), we quantified the number of circulating CD34+ cells (CD34+ cell number) and serum concentration of thrombopoietin (TPO) before and after surgery in 10 donors and 9 recipients undergoing LDLT. The evaluation of CD34+ cells has been carried out by flow cytometry, by applying conventional protocols. In donors, CD34+ cell number increased from 3.69 ± 2.87 cells/μL at pre-LDLT to 8.25 ± 4.94 cells/μL at post-operative day (POD) 7. Platelet count increased from 209 ± 50 × 109/L at pre-LDLT to 273 ± 23 × 109/L at POD 7–11. In recipients, although CD34+ cell number was 0.78 ± 0.73 cells/μL at pre-LDLT and significantly lower than that in healthy volunteers (3.78 ± 3.93 cells/μL), the number at POD 9–42 increased to 4.25 ± 3.55 cells/μL. Platelet count increased from 64 ± 35 × 109/L at pre-LDLT to 197 ± 91 × 109/L at POD 9–42. Serum TPO concentration at pre-LDLT was 0.98 ± 0.61 fmol/L in donors and 1.00 ± 0.35 fmol/L in recipients. TPO level peaked at POD 3–7 (3.14 ± 1.67 fmol/L) in donors and at POD 7–22 (6.74 ± 3.13 fmol/L) in recipients. It has been reported that mobilization of HSCs after partial hepatectomy in donors or LDLT in recipients was likely enhanced by increased serum levels of granulocyte colony-stimulating factor (peaked at POD 1), interleukin-6 and stem cell factor (POD 7). However, in our study, TPO (peaked at POD 7–22) was strongly associated with not only the increase of platelet count (peaked at POD 9–42) but also the mobilization of HSCs (POD 9–42) in recipients. Both mobilized HSCs and platelets may contribute to the liver regeneration after LDLT.
Array-CGH Identifies Regions, Including the FOXP1 Locus, Associated with Different Clinical Outcome in Diffuse Large B-Cell Lymphomas (DLBCL) Treated with R-CHOP
Authors: Scandurra Marta , Rancoita Paola M.V. , Greiner Timothy C. , Chan Wing C. , Vose Julie M. , Inghirami Giorgio , Chiappella Annalisa , Baldini Luca , Ponzoni Maurilio , Ferreri Andres JM , Montes-Moreno Santiago , Piris Miguel A. , Franceschetti Silvia , Gaidano Gianluca , Facchetti Fabio , Tucci Alessandra , Lazure Thierry , Lambotte Olivier , Uccella Silvia , Pinotti Graziella , Pruneri Giancarlo , Martinelli Giovanni , Nomdedeu Josep , Tibiletti Maria Grazia , Chigrinova Ekaterina , De Campos Cassio P. , Rinaldi Andrea , Zucca Emanuele , Kwee Ivo , Bertoni Francesco 
Published: April 2020

Abstract
Background. Diffuse large B-cell lymphoma (DLBCL) consists of a heterogeneous group of tumors. More than 30% of cases are not cured with R-CHOP chemotherapy.
Aim. To identify genomic aberrations that could affect the response to therapy, we performed an arrayCGH study on uniformly treated DLBCL patients.
Materials and Methods. Tumor samples were analyzed with Affymetrix Human Mapping 250K SNP arrays. Eligibility criteria were diagnosis of de novo DLBCL, first line treatment with R-CHOP or R-CHOP-like regimens, availability of frozen biopsy and of clinical baseline and follow-up data. Exclusion criteria were: primary mediastinal DLBCL, primary central nervous system DLBCL, HIV-positivity. Material has been collected according to the local IRB guidelines. Results. Genomic tumor profiles have been obtained in 163 samples from 10 Institutions; 23/163 cases were excluded from this analysis because the information on response was not available; 140/163 cases fulfilled the study requirements. The clinical parameters of the patients reflected the normal DLBLC population, as shown by the IPI score distribution (0–1 in 30%, 2 in 37%, 3 in 23% and 4–5 in 10%). The median follow-up was 23 months (range 1–1251). Complete remission was observed in 108/140 (77%) patients, partial response in 21 (15%), stable disease in 6 (4%) and progression in 5 (2%). Genomic differences were observed between complete responders (108 cases) and the remaining patients, grouped together as poor responders (32 cases). The latter group had more gains of 3p14.1 (FOXP1 locus), 3q29, 11q24.3, and losses of 2p11.2-p13.3, 8p23.1- pter, 10p12.31-p13, 15q11.2-q14, 15q21.1 and copy neutral LOH of chromosome 9p. On the converse, 1q gains were more common among patients achieving complete remission. No differences were observed for other common region of gains (7, 12, 18q/BCL2) or losses (1p, 6q, 17p/TP53).
Conclusions. Specific genomic aberrations are associated with the response to R-CHOP in patients with DLBCL. In particular, the gain of the 3p14.1 (FOXP1 locus), which is associated with a lack of response to R-CHOP, suggests that the role of FOXP1 should be further investigated in DLBCL.
Identification and Characterization of Hematopoietic Stem (HSC)/Progenitor Cells in Human Solid Tumors
Authors: Leung Roland C , Dorn David C , Downey Robert J , Levine Douglas A , Moore Malcolm A.S. 
Published: April 2020

Abstract
It has recently been shown that a solid tumor recruits cells to the tumor mass, which facilitate vascularization, suppress immune responses, aid its growth, and influence metastatic potential. These recruited cells include mesenchymal stem cells, endothelial progenitor cells and tumor infiltrating leukocytes. By performing multiparameter FACS analysis on primary lung carcinoma and pleural effusion material, we demonstrate for the first time the consistent presence of CD45+/CD34+ cells (mean 1.6% of total CD45+cells range 0.24–13.2% n=12). In primary tumor samples, CD45+ lymphohematopoietic cells were purified by the MACS system and examined for progenitor colony formation in methycellulose. Erythroid and myeloid colony formation was observed in the CD45+ selected subfraction. We assayed for HSC activity by cobblestone formation (CAFC) on MS-5 stroma. Week 2 and week 5 CAFC potential was present in CD45+ purified ovarian cancer ascites. In non-irradiated NOD/SCID IL2 gamma null mice injected subcutaneously with dissociated and unfractionated lung tumor and ovarian peritoneal ascites, small numbers of human CD45+ cells were recovered from the bone marrow after 2–3 months which gave rise to hematopoietic colonies. Human CD45+ cells also persisted within the retransplanted subcutaneous tumors. In conclusion, we report the recruitment of HSCs and progenitors to primary tumor sites. The differentiation and self-renewal potential of these HSCs indicate a potential alternative source of tumor-infiltrating leukocytes as opposed to or in addition to recruitment of mature cells from the circulation.
The role of solid tumors in mobilizing HSCs and progenitors from the bone marrow and their selective chemotaxis to the primary and metastatic sites require further investigation. Targeting this novel tumor-HSC interaction may offer new therapeutic opportunities for controlling tumor growth and metastasis.
Nitric Oxide and Vascular Endothelial Growth Factor during Pregnancy in Sickle Cell Disease
Authors: Brittain Julia E. , Ataga Kenneth I , Lee Sean , Thorp John M. , Orringer Eugene P. 
Published: April 2020

Abstract
BACKGROUND: Vascular Endothelial Growth Factor (VEGF) is a multifunctional cytokine that may be clinically relevant in sickle cell disease (SCD). In additional to its role as a potent activator of angiogenesis, VEGF is also a potent inducer of Nitric Oxide (NO) release. Patients with SCD have impaired NO metabolism and decreased bioavailability of NO due to chronic hemolysis which may contribute to the vasculopathy that is present in this disorder. High levels of plasma arginase and the resultant decreased arginine/ornithine ratios have also been proposed to contribute to low NO levels in SCD patients. We have previously reported that pregnant SCD patients have higher circulating levels of the naturally occurring VEGF antagonist, sFLT-1, when compared to pregnant African American controls. High levels of sFLT-1 are associated with poor pregnancy outcomes including preeclampsia and delivery of small for gestational age (SGA) infants.
OBJECTIVE: In this study we wanted to assess the levels of NO, free VEGF, sFLT-1 and arginase activity during pregnancy in SCD. We also sought to explore the relationship between NO and sFLT-1 with pregnancy outcome.
METHODS: We performed an analysis of previously collected plasma/serum samples obtained from the Pregnancy, Infection and Nutrition (PIN) study cohort at UNC- Chapel Hill. Subjects were matched on race, gestational age, BMI and smoking status. Levels of free VEGF were quantified by luminex analysis and confirmed by ELISA. NO was quantified via its metabolite NOx using a modified Griess method in de-proteinated serum/plasma. Arginase activity was quantified by ELISA. Pregnancy outcomes were previously recorded by PIN study personnel.
RESULTS: We found that levels of free VEGF and NO were significantly lower in pregnant women with SCD than in matched pregnant controls (VEGF: 5.8 pg/ml vs. 13.56 pg/ml, p = 0.03, NOx: 14.31 μM vs. 26.99 μM, p=0.0009). We also found a significant inverse correlation between NOx levels and sFLT-1 levels (r2=0.6421, p = 0.0094), but no relationship between NOx and arginase activity in plasma. There was also no difference in arginase activity between pregnant SCD subjects and controls. Finally, pregnant SCD subjects that delivered an SGA infant had significantly lower levels of NOx than those women that did not.
CONCLUSIONS: These results suggest that VEGF may play a role in NO release during pregnancy in SCD. In addition, low NO levels may contribute to poor pregnancy outcomes in pregnant SCD women.
Markers of Coagulation Activation and Inflammation in Sickle Cell Disease and Sickle Cell Trait
Authors: Adam Soheir S. , Zaher Galila , Ibrahim Mahmoud , Ataga Kenneth I , De Castro Laura M. , Key Nigel S. 
Published: April 2020

Abstract
It has been well-described that sickle cell disease (SCD) due to HbSS or HbSC is associated with systemic activation of coagulation. A single study has reported a similar, albeit lesser, degree of chronic hyperactivation of coagulation in subjects with sickle cell trait (SCT) (Am J Hem2002; 69(2):89–94), and we have shown that SCT is a risk factor for venous thromboembolism (Blood2007;110(3):908–12). The aim of this study was to simultaneously compare markers of activation of coagulation and inflammation in SCD and SCT. Ninety Saudi Arabian individuals (40 SCD (HbSS), 20 SCT (HbAS) and 30 controls (HbAA)) were recruited. SCD patients were not in vaso-occlusive crisis at the time of sample collection. None of the subjects was taking any relevant medication (anticoagulants, hormones or hydroxyurea), nor had they received blood transfusion for at least 6 weeks. On all subjects, complete blood count (CBC), prothrombin time (PT), activated partial thromboplastin time (APTT), D-dimer antigen, protein C activity, protein S activity, antithrombin (AT) activity, fibrinogen concentration, activated protein C resistance ratio (APCR), and C-reactive protein (CRP) levels were obtained. Student’s “t” test was used to compare two independent means. Multivariate analysis “ANOVA” test was done to test statistical difference between more than 2 means.
Results: D-dimer and CRP were significantly higher, whereas protein C, protein S, and fibrinogen were significantly lower in SS patients compared to controls (Table). In SCT, D-dimer levels were higher, and levels of proteins C and S were significantly lower than controls. APC resistance was more pronounced in both SCD and SCT compared to controls. In summary, this study provides further evidence of hypercoagulability in SCD, but more intriguingly, confirms that significant changes in coagulation parameters may be found in SCT. We speculate that loss of asymmetry of phospholipids in the red cell membrane may occur in certain hypoxic vascular beds (such as the renal vasculature) in SCT. We are currently exploring other possible underlying mechanisms that may contribute to activation of coagulation in SCT.
Table: Comparison of Markers of Inflammation and Coagulation Activation in all Groups
Group I (SS) (n=40) Group II (AS) (n=20) Group III (AA) (n=30) *P &lt; 0.05 comparing SCD and control groups by paired t- test **P &lt;0.05 comparing SCT and control groups by paired t- test CRP 22.5 ± 10.9* 0.3 ± 0.2 0.3 ± 0.1 PT 12.5 ± 1.6* 12.2 ± 1.0** 11.5 ± 0.6 APTT 33.3 ± 4.2 35.8 ± 4.6** 32.9 ± 3.2 D-dimer 287 ± 140* 278 ± 207** 141 ± 65 Protein C 77.9 ± 20.3* 97.7 ± 21.7** 109.4 ± 17.2 APCR 0.82 ± 0.12* 0.79 ± 0.12** 1.05 ± 0.19 Protein S 54.0 ± 18.4* 64.6 ± 15.4** 89.7 ± 14.9 AT 94.4 ± 16.6* 102.2 ± 7.4** 109.3 ± 10.7 Fibrinogen 305 ± 101 361 ± 146 358 ± 80