bluuebunny/crossref_metadata_2025_split huggingface dataset:
Biological Significance of Erythropoietin and Erythropoietin Receptor Expression by Human Ovarian Cancer Cells.
Authors: Jeong Jee-Yeong , Feldman Laurie , Solar Peter , Sytkowski Arthur J. 
Published: April 2020

Abstract
Erythropoietin (Epo) is the primary regulator of erythroid cell proliferation and differentiation mediated through its specific binding to the Epo receptor (EpoR). Epo also is known to have non-hematopoietic actions, including the promotion of wound healing, tissue protection and others. Accumulating evidence demonstrating both Epo and EpoR expression in many types of human cancers has raised concern about the clinical use of Epo and other erythropoiesis stimulating agents (ESAs) in cancer-related anemia, since the presence of functional EpoR on the tumor cells suggests the potential for an adverse effect of Epo treatment by enhancing cancer growth and progression. Previously, we demonstrated that A2780 ovarian cancer cells express the EpoR gene and that long-term Epo treatment of these cells renders them resistant to paclitaxel. We now report the characterization and biological significance of Epo and EpoR expression by four ovarian cancer cell lines. Using semi-quantitative RT-PCR, restriction digestion of the PCR products and DNA sequence analysis, we demonstrated that A2780, CaOV, SKOV, and OVCAR-3 ovarian cancer cell lines examined express Epo and EpoR at the mRNA level. We demonstrated EpoR protein expression both by western blotting and by immunofluorescence, and demonstrated biologically active Epo protein expression by quantitative in vitro bioassay of the cell culture supernatants. The EpoR on A2780 cells are functional, since Epo stimulation of the cells resulted in a time-dependent 5-fold increase in phosphorylation of Erk1/2 that reached a peak in 10 min and then returned to the basal level. None of the cell lines exhibited a short-term (3-day) growth response in culture to exogenous Epo. However, addition of a neutralizing anti-Epo antibody to the cell culture resulted in partial growth inhibition of A2780 cells that was reversed by addition of excess Epo, consistent with an autocrine/paracrine mechanism of Epo growth enhancement of these cells. We also found that long-term Epo treatment of A2780 cells resulted in the development of a phenotype exhibiting enhanced Epo signaling, evidenced by a 20-fold increase in phosphorylation levels of Erk1/2. This phenotype was sustained even after the removal of Epo, suggesting a mechanism underlying the paclitaxel resistance developed after long-term Epo treatment. Our findings have implications for the clinical use of recombinant human Epo and other ESAs to correct and/or prevent anemia in ovarian and, potentially, other cancer patients. They also suggest that endogenous Epo and the EpoR may serve as novel therapeutic targets in the treatment of cancer patients.
Protective Role of Connexin 32 in Experimental Leukemogenesis.
Authors: Hirabayashi Yoko , Yoon Byung-Il , Tsuboi Isao , Huo Yan , Kodama Yukio , Kanno Jun , Ott Thomas , Trosko James E. , Inoue Tohru 
Published: April 2020

Abstract
Connexin (Cx) functions in the organization of cell-cell communication in multicellular organisms. Gap junctions have been implicated in the homeostatic regulation of various cellular functions, including growth control and differentiation, apoptosis, and the synchronization of electrotonic and metabolic functions. Primitive hemopoietic progenitor cells form a multicellular system, but a previous report describes that Cx32 is not expressed in the bone marrow. Thus, a question arises as to why Cx molecules are not detected in the hematopoietic tissue other than stromal cells. Based on our preliminary study that suggested a potential impairment of hematopoiesis in Cx32-knockout (KO) mice, the objectives of the present study were to determine whether Cx32 functions in the bone marrow during steady-state hematopoiesis and further to examine its possible protective roles during regeneration after chemical abrasions and during leukemogenesis after the administration of a genotoxic chemical, methyl nitrosourea (MNU). As results, the Cx32 molecule functioning in the hematopoietic stem cell (HSC) compartment during steady-state hematopoiesis was observed for the first time; the expression of Cx32 at the mRNA level determined by PCR analysis and that at the protein level determined using an anti-Cx32 antibody were observed only in the lin−c-kit+ HSC fraction using a combination of immunobead-density gradient and immunomagnetic-bead separation. Hematopoiesis was impaired in the absence of Cx32; it was delayed during regeneration after chemical abrasion with 5-fluorouracil at 150 mg/kg body weight in Cx32-KO mice. Cx32-KO mice also showed increased leukemogenicity compared with wild-type mice after MNU injection; furthermore, in a competitive assay for leukemogenicity in mice that had been lethally irradiated and repopulated with a mixed population of equal amount of bone marrow cells from Cx32-KO mice and wild-type mice, the resulting leukemias were originated predominantly from Cx32-KO bone marrow cells. The present competitive assay clearly showed that Cx32-KO bone marrow cells have a higher risk of becoming leukemogenic. The above-mentioned findings in this study imply that Cxs play an essential role in maintaining the steady-state hematopoiesis and suppressing the neoplastic change. In summary, the role of Cx32 in hematopoiesis was not previously recognized and Cx32 was expressed only in HSCs and their progenitors. The results indicate that Cx32 in wild-type mice protects HSCs from chemical abrasion and leukemogenic impacts. Our results indicate that the risk of developing leukemia in patients with X-chromosome-linked Cx32 deficiency, called Charcot-Marie-Tooth syndrome, may not be incidental.
Transcription Factor Dosage: Maximal STAT5-Induced Proliferation and Stem Cell Self-Renewal at Intermediate STAT5 Activity Levels.
Authors: Wierenga Albertus T.J. , Vellenga Edo , Schuringa Jan Jacob 
Published: April 2020

Abstract
The level of transcription factor activity critically regulates cell fate decisions such as hematopoietic stem cell self-renewal and differentiation. Here, we describe a model in which STAT5 activity is introduced into human hematopoietic stem/progenitor cells in a dose-dependent manner by overexpression of tamoxifen-inducible STAT5A-ER fusion proteins that can be activated upon treatment with 4-OHT. For this study, we tested STAT5A(wt), STAT5A(1*6) and STAT5A(S711F)-ER fusion proteins. Introduction of STAT5A activity in cord blood-derived CD34+ cells resulted in impaired myelopoiesis and induction of erythropoiesis which was most pronounced at the highest STAT5A transactivation levels. In contrast, intermediate STAT5A activity levels resulted in the most pronounced proliferative advantage of CD34+ cells. This coincided with increased CAFC and LTC-IC frequencies that were predominantly elevated at intermediate STAT5A activity levels, but not at high STAT5A activity levels. Self-renewal of progenitors was addressed by serial replating of CFUs grown in methylcellulose, and only progenitors containing intermediate STAT5A activity levels contained self-renewal capacity. Extensive gene expression profiling in CB CD34+ cells over a range of STAT5A activation levels identified subsets of genes that were induced by low, intermediate and high STAT5A transcriptional activity. Thus, we could cluster gene expression levels of STAT5 target genes into a ‘differentiation signature’ versus a ‘proliferation/self-renewal signature’. In conclusion, our data suggest that intermediate rather than high STAT5 activity levels might be required to induce leukemic transformation.
Ligation of TLR5 by Flagellin Converts Tolerogenic Dendritic Cells into Activating Dendritic Cells through Inhibition of IL-10 Production.
Authors: Vicente-Suarez Ildefonso , Sotomayor Eduardo M. 
Published: April 2020

Abstract
Antigen presentation by DCs differentiated in the presence of the anti-inflammatory cytokine IL-10 has been shown to result in the development of regulatory T cells (Tregs) and tolerance induction. Whether this outcome could be reverted -or not- through manipulation of the tolerogenic DCs (tol-DCs) has not been explored in detail. Here we report for the first time that tol-DCs differentiated in the presence of IL-10 display increased expression of TLR5 as compared with bone marrow-derived DCs differentiated in the presence of GM-CSF and IL-4 (BM-DCs). The increased expression of TLR5 in tol-DCs seems to be a unique characteristic given that expression of other TLRs such as TLR4 and TLR9 was not different in tol-DCs as compared to BM-DCs. In lieu of our previous observation that the TLR5 ligand, flagellin, inhibits IL-10 production in antigen-presenting cells (APCs)1, and the unexpected finding that tol-DCs display higher expression of TLR5, we asked next whether flagellin could influence the functional properties of tol-DCs. First, unlike BM-DCs, tol-DCs stimulated with LPS (TLR4 ligand) produced high levels of IL-10. In sharp contrast, tol-DCs stimulated with flagellin do not produce IL-10 but produce significant levels of the pro-inflammatory cytokine IL-12. Second, while antigen presentation by LPS-stimulated tol-DCs results in the generation of IL-10 producing CD4+ T-cells, presentation of cognate antigen by flagellin-stimulated tol-DCs resulted in CD4+ T-cells that secrete IFN-gamma but not IL-10. This divergent T-cell outcome in response to antigen-presentation by tol-DCs stimulated with LPS (IL-10 producing T-cells) versus flagellin (IFN-gamma producing T-cells) suggests that TLR5 engagement might be a novel approach to convert tolerogenic DCs into activating DCs and effectively trigger productive immune responses in environments that are otherwise conducive to unresponsiveness due to the presence of IL-10.
Chemotherapy Alone Versus Chemotherapy Plus Radiotherapy for Early Stage Hodgkin Lymphoma.
Authors: Rehan Fareed A. , Brillant Corinne , Schulz Holger , Knauel Ina , Bohlius Julia , Specht Lena , Engert Andreas 
Published: April 2020

Abstract
Both chemotherapy (CT) alone and combined modality therapy (CMT) are effective modalities for the treatment of early stages of Hodgkin lymphoma (HL). However, the optimal choice of treatment is still being discussed. Different research groups reported that ABVD-like CT followed by radiotherapy is highly effective for early favourable and early unfavourable (intermediate) stage HL. A recently conducted randomized trial aimed at determining whether CMT is superior to CT alone in early stages, showed no significant difference in 5 year overall survival (OS) and event-free survival in patients treated with either 4–6 cycles of ABVD alone or 2 cycles of ABVD plus radiotherapy. Thus, this systematic review and meta-analysis was performed for a more comprehensive comparison.
Methods: Only randomized controlled trials (RCT) comparing CT alone with CMT in newly diagnosed adults with early stages of HL (CS IA, IB, IIA, IIB) were included. Medline and Cochrane Library were systematically searched for randomized controlled trials from 1975 to 2007. Patients who received CMT were considered as experimental group and patients who received ABVD-like CT alone were considered as control group. Data were collected from full text publications or abstracts and treatment effects for OS were calculated as hazard ratios (HR).
Results: A total of 590 references were screened. Four eligible RCTs were identified, including 1207 patients with early-stage HL (early favourable and early unfavourable stages). Between 1983 and 2004, 765 patients where treated with CMT and 442 patients with CT alone. The OS for the CMT group was significantly better than the OS for the CT-alone group (HR: 0.54; 95% CI [0.35 - 0.84]; p = 0.006), but there was evidence for a substantial heterogeneity between the studies (I2 = 69%, p=0.02).
Discussion: First results of this systematic review showed improved OS for patients treated with CMT in early-stage HL compared to patients treated with CT alone. The substantial heterogeneity between the trials should be clarified. Further comprehensive analyses are ongoing and will provide more detailed information.
Does FDG-PET/CT Obviate the Need for Standard Diagnostic Contrast-Enhanced CT in Patients with Lymphoma?.
Authors: Elstrom Rebecca L. , Brown Richard K.J. 
Published: April 2020

Abstract
Background and Significance: Positron emission tomography using 18-fluoro-2-deoxyglucose (FDG-PET) is useful in the staging and follow up of patients with lymphoma, and has been shown in several studies to be more accurate than computed tomography (CT). These studies have, however, demonstrated a continued role for CT in staging and restaging of lymphoma, and the two modalities are complementary. Increasingly, FDG-PET is performed in conjunction with a low radiation dose, non-contrast CT scan for attenuation correction and localization of lesions. Currently, many patients undergo both FDG-PET/CT and standard diagnostic, contrast enhanced CT, at a significant cost both financially and in terms of radiation exposure. In this study, we evaluated the clinical utility of performing both studies in patients with lymphoma.
Study Design and Methods: We retrospectively identified patients with lymphoma who had undergone both FDG-PET/CT and diagnostic, contrast-enhanced CT (a scan pair) for either staging or restaging following treatment. Patients were included if the two imaging studies were performed within 6 weeks of each other with no intervening anti-lymphoma therapy. We compared the results of the two studies, identifying findings that were detected in either FDG-PET/CT or diagnostic CT scan but not both. Discrepancies were considered clinically significant if they were determined to be related either to lymphoma or another disease process which potentially required intervention.
Results: Eighty-nine scan pairs which met the criteria were identified in 75 patients. Sixty-one scan pairs were performed for staging, and 28 were performed for treatment follow up. FDG-PET/CT detected additional potentially clinically relevant lesions over CT in 30 patients, of which 11 demonstrated increased clinical stage. Lymphoma therapy was changed based on FDG-PET/CT findings in 2 patients, and in one patient an occult rectal cancer was detected. In contrast, diagnostic CT detected 5 potentially clinically relevant findings, including 2 incidental findings (one definite and one possible venous thrombosis), and 3 patients with splenic lesions. Of the patients with splenic lesions, one was found on follow up to be definitely not related to lymphoma, and the nature of the splenic lesions in the other two patients remained indeterminate. No patient had a change of stage or lymphoma therapy based on diagnostic CT scan, and one patient was treated with anticoagulation based on CT findings. In the subgroup of scan pairs performed for follow up, diagnostic CT added clinically relevant information in none of the patients.
Conclusion: In our series of patients, diagnostic contrast-enhanced CT scan did not contribute to staging or restaging of lymphoma when performed concurrently with FDG-PET/CT. Two clinically important incidental findings were detected by CT alone, of which one led to intervention.
Case-Control Study Using Array-Based Genotyping Produced a Panel of Genes Associated with Risk to Acute Myelogenous Leukemia (AML).
Authors: Lee Il-Kwon , Kim Hee Nam , Kim Yeo-Kyeoung , Park Kyeong-Soo , Yang Deok-Hwan , Lee Je-Jung , Shin Myung Geun , Choi Chan , Kook Hoon , Hwang Tae Ju , Park Moo Rim , Jo Deog Yeon , Won Jong Ho , Kim Hyeoung-Joon 
Published: April 2020

Abstract
Multiple loci with small genetic effects are thought to be linked to AML pathogenesis. To detect such loci requires systematic screening of large number of single nucleotide polymorphisms (SNPs) within large study population. In this study we tested whether 3104 SNPs in 200 candidate genes were associated with risk to AML. Genes were considered potential candidates for their known or suspected roles in DNA repair system, pharmacogenomics and transcriptional regulation in hematopoiesis or putative pathways related to leukemogenesis. Selection of SNPs was performed using dbSNP and HapMap project databases, with emphasis on non-synonymous SNPs or haplotype tagging SNPs. To discover potential SNPs responsible for AML susceptibility, we conducted a case-control study using Affymetrix targeted genotyping 3K array. We applied this platform to analyze the samples from 309 de novo AML patients and 382 healthy controls. Genotype scorings were processed using GeneChip scanner 3000 TG(Affymetrix) and analyzed with GCOS software(Affymetrix). In total more than 255,000 SNPs were genotyped for this study. Samples with suboptimal call rates were excluded. Statistical testing were carried out using χ2, Cochran-Armitage trend, Fisher’s exact, odds ratio, haplotype estimation, LD block definition. Here we report that 23 SNPs in 16 genes are associated with elevated or reduced risk to AML. Some of associated genes were transcription factors such as ZNF23 and ZNF233. While associated genes were distributed evenly among the whole genome, four associated genes were found on chromosome 1. Among those associated SNPs, two were located in coding region, one in exon-intron boundary, while the rest of SNPs were located in introns of associated genes. Among 23 SNPs identified, two intronic SNPs from 1st intron of BAALC(Brain and acute leukemia, cytoplasmic) gene were associated with the reduced risk to AML. Haplotype estimation and linkage disequilibrium pattern of 9 SNPs including two associated SNPs will be presented. In conclusion, we identified SNPs responsible for AML susceptibility by candidate gene-based SNP array approach. These promising data when supported by further molecular validation would greatly enhance the current understanding of AML predisposition and diseases progression. Implication of polymorphic variants in AML etiopathogenesis will be presented and discussed.
Molecular Allelo-Karyotyping of Adult T-Cell Leukemia Using High SNP Genotyping Microarrays.
Authors: Muto Satsuki , Yamamoto Go , Nannya Yasuhito , Sanada Masashi , Dabaghmanesh Nazanin , Ishida Takaomi , Utsunomiya Atae , Yamada Yasuaki , Kamihira Shimeru , Yamaguchi Kazunari , Watanabe Toshiki , Ogawa Seishi 
Published: April 2020

Abstract
Adult T cell leukemia/lymphoma (ATL) is a mature T-cell neoplasm in adult that is caused by human T-cell leukemia virus type 1 (HTLV-1) and highly intractable to conventional therapeutics. Since there are 1.2 million HTLV-1 carriers in Japan and more than 50,000 carriers are expected to develop ATL from now on, it is of particular importance to understand the pathogenesis of ATL. The malignant processes of T-cell transformation in ATL are initiated by HTLV-1 infection in early childhood, and the HTLV-1 infected and immortalized T-cells are thought to accumulate a series of genetic hits during the later life, ultimately giving rise to malignant ATL clones after decades of latency periods. However, little is known about the nature of these genetic hits that take place after the early immortalization processes of T-cells mediated by HTLV-1 Tax protein. So in order to clarify the genetic alterations involved in the later processes of T-cell transformation in ATL, we analyzed a total of 130 ATL samples using Affymetrix® GeneChip® 250K Nsp arrays. Combined with CNAG/AsCNAR software, these arrays allow for accurate determination of allele-specific copy numbers in extremely high-resolution (less than 12kb) to detect copy number alterations as well as allelic imbalances in ATL genomes without depending on the availability of constitutive DNA of tumor specimens (molecular allelo-karyotyping). ATL genomes show characteristic copy number profiles and unique patterns of allelic imbalances, which are distinct from acute lymphoblastic ALL and non-Hodgikin’s lymphomas and include gains of 1q arm, 2q33, 3p and 3q arms, 9p12, 17q12, and 19p13, and losses of 1p13.1, 2q end, 3q22, 4q31, 6p22, 7q31, 9p21, 10p14, 12q13, 14q24, and 19q13. Moreover, allele-specific determination of copy numbers disclosed a number of regions showing copy number neutral LOH. Numerous homozygous deletions and gene amplifications were also identified and commonly mapped to less than 500Kb regions, which facilitated identification of candidate gene targets. Interestingly, these genetic lesions involved many T-cell related genes, indicating that the de-regulation of normal T cell functions may contribute the pathogenesis of ATL. In conclusion, molecular allelo-karyotyping of ATL genomes using SNP arrays provides valuable information about the molecular targets in ATL pathogenesis.
Gas6 and Its Receptors Are Implicated in Sepsis as Modulators of Innate Immunity.
Authors: Burnier Laurent , Sugamele Rocco , Le Roy Didier , Roger Thierry , Fumeaux Thierry , Chanson Marc , Borgel Delphine , Lemke Greg , Matsushima Glenn K. , Earp H. Shelton , Carmeliet Peter , Schapira Marc , Feihl François , Calandra Thierry , Angelillo-Scherrer Anne 
Published: April 2020

Abstract
Gas6 downregulates the activation state of macrophages and thereby their production of proinflammatory cytokines induced by various stimuli. We aimed to determine whether Gas6 is involved in sepsis. We measured Gas6 plasma levels in 13 healthy subjects, 29 patients with severe sepsis, and 18 patients with non-infectious inflammatory diseases. Gas6 level was higher in septic patients than in control groups (P≤0.0001). The sensitivity and specificity of Gas6 levels to predict fatal outcome were 83% and 88%. We next investigated whether Gas6 affects cytokine production and outcome in experimental models of endotoxemia and peritonitis in wild-type (WT) and Gas6−/− mice. Circulating levels of Gas6 after LPS 25mg/kg i.p. peaked at 1 hour (P&lt;0.001). Similarly, TNF-α was higher in Gas6−/− than in WT mice 1 hour after LPS (P&lt;0.05). Furthermore, 62 anti- and pro-inflammatory cytokines were quantified in plasma after LPS injection. Their levels were globally higher in Gas6−/− plasma after LPS, 47/62 cytokines being at least 50% higher in Gas6−/− than in WT plasma after 1 hour. Mortality induced by 25mg/kg LPS was 25% in WT versus 87% in Gas6−/− mice (P&lt;0.05). LPS-induced mortality in Gas6 receptors Axl−/−, Tyro3−/− and Merkd was also enhanced when compared to WT mice (P&lt;0.001). In peritonitis models (cecal ligation and puncture, CLP, and i.p. injection of E. coli), Gas6 plasma levels increased and remained elevated at least 24 hours. CLP increased mortality in Gas6−/− mice. Finally, we explored the role of Gas6 in LPS-treated macrophages. We found that Gas6 was released by LPS-stimulated WT macrophages and that Gas6−/− macrophages produced more TNF-α and IL-6 than WT macrophages. Cytokine release by Gas6−/− macrophages was higher than by WT macrophages (cytokine array). Adjunction of recombinant Gas6 to the culture medium of Gas6−/− macrophages diminished the cytokine production to WT levels. In LPS-treated Gas6−/− macrophages, Akt and Erk1/2 phosphorylation was reduced whereas p38 and NFκB activation was enhanced. Thus, in septic patients, elevated Gas6 levels were associated with fatal outcome. In mice, they raised in experimental endotoxemia and peritonitis models, and correlated also with sepsis severity. However, Gas6−/− mice survival in these models was reduced compared to WT. Gas6 secreted by macrophages in response to LPS activated Akt and restrained p38 and NFκB activation, thereby dampening macrophage activation. Altogether these data suggest that, during endotoxemia, Gas6−/− mice phenotype resembles that of mice which have undergone PI3K inhibition, indicating that Gas6 is a major modulator of innate immunity.