bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mesenchymal Stem Cells (MSC) Exhibit Antigen Presenting (APC) and Phagocytic Properties: Implicatins to Bnone Marrow Failure during Inflammation.
Authors: Chan Jennifer L. , Harrison Jonathan S. , Ponzio Nicholas M. , Rameshwar Pranela 
Published: April 2020

Abstract
Mesenchymal stem cells (MSC) mostly surround the vasculature system of bone marrow (BM). MSC have been shown to exhibit immune suppressive properties. Since MSC express MHC Class II antigen, the question is whether these cells can act as APC. To this end, we hypothesize that MSC have the ability to present non-self antigens while acting as immune modulators. These dual roles of MSC prevent exacerbated inflammatory responses in the BM, thereby preventing hematopoietic dysfunction.
A ‘dampened’ immune response in BM during insults by foreign agents could cause protection of the barrier that separates BM cavity with the periphery. The phagocytic role of MSC was shown by confocal microscopy and fluoresbrite plain YG 1.0-micron microspheres. APC property was demonstrated by challenging MSC with C. albicans (pulsed MSC), followed by exposure to CD4+ cells. The latter was obtained by immunoselection from peripheral blood mononuclear cells (PBMC) cultured for 5 days with C. albicans (10 mg/ml). Proliferation of the CD4+ cells (3H-thymidine incorporation and cell counts) proved APC properties of MSC, at efficiency comparable to macrophages. Overall, the studies show that the window between APC function and the period at which MSC could become immune suppressive is critical, since activated T-cells could destroy the endothelial barrier between BM and lymphatics/peripheral circulation. These studies show that MSC could be key cells in regulating immune responses in BM, and thereby protect BM from failure.
IFNg-Mediated MHC-II Expression in Mesenchymal Stem Cells (MSC) Is Concentration-Dependent.
Authors: Tang Katherine C. , Kotenko Sergei , Harrison Jonathan S. , Rameshwar Pranela 
Published: April 2020

Abstract
A significant concern in tissue repair is allo-rejection in a host with MHC-II mismatch. Since MSC appear to have veto properties to offset its own rejection, these stem cells may serve as ideal candidates in repair medicine. Interferon-g (IFN-g), which exacerbates immune responses, is known to enhance the expression of MHC-II in antigen presenting cells (e.g., macrophages). This led to the question on the mechanism by which MSC act as immune suppressor cells. We hypothesize that endogenous IFN-γ maintains basal MHC-II expression, while increased levels lead to its down regulation. Indeed, we observed a biphasic response of IFN-γ on the expression of MHC-II. While unstimulated MSC produced 19±5 pg/ml IFN-γ, exogenous IFN-γ (&gt;100 U/ml) led to decreased expression of MHC-II. Suppression of IFN-g by siRNA resulted in undetectable MHC-II. The response of exogenous IFN-γ could not be explained by altered expression of IFN-g receptor type I (IFN-γRI) or changes in STAT-1 activation. However, western blots showed multiple isotypes of the MHC-II master regulator transcription factor, CIITA, in IFN-γ-stimulated MSC, with predominance of CIITA exhibiting low efficiency binding to DNA. This observation might explain why MHC-II is undetectable in the presence of high levels of IFN-γ. These findings support a potential for MSC as an adult stem cell in tissue repair.
Bone Marrow Stroma-Derived Mesenchymal Cell Cultures from Nude/Nude Rat Showing Both Ex Vivo Differentiation Properties and High Metastatic Potential in Nude Mice.
Authors: Wang Mengyu , Kvalheim Gunnar , Mikalsen Svein-Ole , Nesland Jahn M. , Mælandsmo Gunhild , Fodstad Øystein 
Published: April 2020

Abstract
Recent studies have revealed that stem cell like cancer cells can be isolated from different types of primary tumor tissue from brain, breast and skin. Such cells have high capability to form tumors when transplanted into experimental animals. Here we describe a newly developed nude rat mesenchymal stem cellS (rMSCs), which spontaneously changed in vivo properties during culturing. Rat bone marrow cells were isolated from the femur and tibia of a male nude/nude rat and cultured in DMEM medium supplemented with 20% bovine fetal serum (FCS). Non- adherent cells were removed after 72 hours of incubation and the adherent bone marrow cells were cultured until a confluent layer appeared. Adherent cells were detached by trypsin, washed and continued to be cultured. After 23 passages by high-density culture the rMSCs spontaneously became more homogenous and formed spheroids in three-dimensional culture. Following ex vivo culturing under different growth conditions these cells were still able to differentiate into fat and neural like cells. The rMSCs also contain high numbers of colony-forming-unit-fibroblasts(cfu-f). The “tumourigenic” growth pattern observed ex vivo prompted us to examine the rMSCs in vivo. Following subcutaneous injection of 1 x106 cells into nude/nude mice a solid tumor appeared under the skin after 12 days. Moreover, intravenous injections and direct injections into left heart ventricle of rMSCs created solid tumors both in the lung, abdominal cavity, bone and skin. Both phenotypic and genetic characterization of early and late passages of the rMSCs and the tumors formed in the mice are being investigated and more details will be presented at the meeting.
Co-Activation of HOXB7 and BCL2/MYC Via Biallelic IGH Rearrangements in a B-Cell Lymphoma Cell Line.
Authors: Nagel Stefan , Kaufmann Maren , Drexler Hans G. , MacLeod Roderick A.F. 
Published: April 2020

Abstract
Homeobox (Hox) genes mediate tissue-specific expression during development where dysregulation may provoke neoplasia and the involvement of specific Hox genes in hematopoietic malignancies is a recurring theme. In B-cell neoplasias dysregulation may be effected by juxtaposition of Hox genes with immunoglobulin gene regulatory elements, notably IGH. We describe the molecular cytogenetic analysis of a cell line (Epstein-Barr virus negative) established from ascites fluid taken from a 67-year-old male patient at diagnosis of B-cell non-Hodgkin’s lymphoma (follicular, small cleaved cell type). The cell line carries a complex t(8;14;18)(q24;q32;q21) leading to activation of both MYC (at 8q24) and BCL2 (at 18q21) via juxtaposition with different enhancer regions of the IGH locus at 14q32. Both translocated alleles of MYC and BCL2 showed additional genomic amplification in conjunction with IGH by formation of an homogeneously staining region on the der(8). Interestingly, the IGH allele on the second chromosome 14 homolog had also undergone chromosomal juxtaposition, this time with a breakpoint at the HOXB cluster at 17q21 leading to transcriptional activation of HOXB7 but of no other HOXB gene. Although its deregulated expression has been described in certain solid tumor subtypes, notably malignant melanoma, this is the first report describing the involvement of HOXB7 in hematopoietic malignancy. Moreover, the cell line carries a novel internal duplication of chromosome 3, dup(3)(q21q27), effecting fusion of BCL6 with MBNL1, a triplet-expansion repeat gene preferentially expressed during hematopoiesis. Our data thus identify a cell line resource modelling novel hematopoietic rearrangements, notably HOXB7 - an attractive candidate target of recurrent 17q2 rearrangements in B-cell lymphoma.
Decrease of Malignant Potential of t(8;21) Positive Cells after Stable Expression of RUNX-CBFA2T1-Specific Small Interfering RNA.
Authors: Drescher Bettina , Goerlich Kerstin , Reile Dagmar , Heil Gerhard , Heidenreich Olaf , Wagner Katharina , Ganser Arnold , Krauter Juergen 
Published: April 2020

Abstract
The reciprocal chromosomal translocation t(8;21) results in the fusion of the DNA-binding domain of the transcription factor RUNX1 (also called AML1 or CBFα) to CBFA2T1 (also called MTG8 or ETO). The chimeric protein RUNX1-CBFA2T1 interferes with hematopoietic gene expression by recruiting histone deacetylases via N-CoR and mSin3. In addition, impairment of the function of transcription factors such as C/EBPα has been described. To further investigate the function of RUNX1-CBFA2T1 in the development of leukemia, the expression of the fusion protein was inhibited by small interfering RNAs (siRNAs). For stable expression of the siRNA in the target cells, a RUNX1-CBFA2T1-specific siRNA-sequence was cloned into a lentivial vector as short-hairpin-RNA (shRNA). Subsequently, the t(8;21) positive Kasumi-1 cell line was infected with an effeciency of &gt;95% as detected by expression of the GFP reporter gene. As a control, an shRNA targeting luciferase mRNA was used. Expression of the anti-RUNX1-CBFA2T1-shRNA in the Kasumi-1 cells led to a marked reduction of mRNA and protein expression of the fusion gene, whereas the expression of the wildtype RUNX1 gene was not affected. The surface expression of the M-CSF-Receptor, which is a known target gene of C/EBPα, was increased in the RUNX1-CBF2T1 depleted cells. Also, Kasumi-1 cells treated with the shRNA displayed a decrease in CD34 surface expression. In parallel, CD34 mRNA expression was reduced to 10%. To analyze, if CD34 downregulation of the Kasumi-1 cells after RUNX1-CBF2T1 depletion correlates with a loss of progenitor status, the clonogenicity of the cells in semisolid medium was investigated. In Kasumi-1 cells treated with the shRNA against RUNX1-CBFA2T1 the number of spontaneous colonies after 14 days of culture was reduced to about 30% in comparison to cells expressing the control shRNA. In conclusion, these experiments show that RUNX1-CBF2T1 expression can be stably suppressed in t(8;21) positive cells by endogenously expressed shRNAs thereby reducing their clonogenic potential.
A Novel Fusion, KIAA0379 (Ankyrin Repeat Domain 28) to NUP98 in an Adult MDS/AML with a t(3;5;11)(p25;q35;p15).
Authors: Ishikawa Maho , Yagasaki Fumiharu , Okamura Daisuke , Maeda Tomoya , Sugahara Yuichi , Misumi Motohiro , Yoshida Katsuhiko , Nakamura Yuichi , Kawai Nobutaka , Matsuda Akira , Jinnai Itsuro , Bessho Masami 
Published: April 2020

Abstract
In hematologic malignancies, deletion of 3p25-26 is a rare but recurrent cytogenetic aberration, indicating the presence of tumor suppressor gene (TSG) on this choromosome arm.
We revealed a t(3;5)(p25;q35) that was identified by conventional cytogenetics results in a novel fusion of KIAA0379 on 3p25 to NUP98 on 11p15 in a patient with refractory MDS/AML by the molecular analysis. Fluorescence in situ hybridization (FISH) analysis using whole chromosomal painting and telomeric probes unclosed a criptic three-way translocation t(3;5;11)(p25;q35;p15). FISH restriction using BAC contig revealed that the 3p25 breakpoint is involved in KIAA0379 which locates between BAC 792N12 and 259O19. Split signals of BAC 265K23 for 5q35 and 348A20 for 11p15 showed that NSD1 on 5q35 and NUP98 on 11p15 are the responsible genes of this translocation. As the results of this translocation, expression of three chimeric transcripts were identified by RT-PCR. In KIAA0379-NUP98, exon 18 of KIAA0379 was fused in-flame with exon 13 of NUP98. In NUP98-NSD1, exon 12 of NUP98 was fused in-flame with exon 7 of NSD1. NSD1-KIAA0379 resulted in out-flame fusion. To investigate the expression of KIAA0379 in leukemic cells, RT-PCR for full-length KIAA0379 transcript was performed on bone marrow cells from the patient and a healthy volunteer, and leukemic cell line K562, HL-60 and KG-1a cells. Full-lengh PCR product with expected size was found in all samples, however, short variant transcript was amplified significantly from KG-1a and HL-60 cells. Subsequent sequence analysis of the short PCR product from KG-1a cells revealed a direct fusion of a part of exon 10 to a part of exon 28 without any hidden splicing sites. Southern blot analysis using a cloned KIAA0379 cDNA fragment suggested an internal deletion of KIAA0379 in KG-1a cells. KIAA0379 locates at 3p25.1 about 5.5Mb centromeric to VHL and is predicted to have 28 ankyrin repeat motifs by computational analysis. It is thought to be involved in protein to protein interaction whereas its precise funtion remains unclear. Immunostaining for KIAA0379 protein in NIH3T3 cells transfected with the wild KIAA0379 and the short variant KIAA0379 revealed that the wild KIAA0379 protein localized in cytoplasm and nuclear membrane, on the other hand, the truncated KIAA0379 protein localized exclusively on nuclear membrane. KIAA0379-NUP98 fusion protein is expected to localize on nuclear membrane by sub-localization signal of C-terminal of NUP98. Therefore, dysfunction of KIAA0379 protein might lead to leukemogenesis and KIAA0379 may be potential candidate for a novel TSG. In addition, it has been reported that t(5;11)(q35;p15.5) results in NUP98-NSD1 fusion in childhood AML. To our knowledge, this is the first report in adult case.
Experimental Design & Analysis of Protein Array Data: Applying Methods from cDNA Arrays.
Authors: Eckel Jeanette E. , Hoering Antje , Ghobrial Irene 
Published: April 2020

Abstract
It appears that a number of recent manuscripts using protein microarray technology are using equivalent analysis procedures that the gene-expression microarray community implemented in their infancy. That is, utilizing a classic reference design such that the ratio of the sample of interest to a reference sample is the response of interest and assessing fold change to determine differential expression. For example, recent publications have concluded that proteins with a fold change less than 0.7 or greater than 1.3 demonstrate significant down- or up-regulated differential expression, respectively. However, fold change is an unreliable measure of differential expression and statistical models that distinguish true signal from random noise should be utilized instead of fold changes. Over the last half decade a tremendous amount of research has been devoted to gene-expression microarrays to vastly improve on the areas of experimental design, normalization and statistical analyses to assess differential expression and classification and these methods are directly applicable to protein microarray technology. Thus, the objective is to review the statistical methodology that has been developed for two-color cDNA arrays that is directly applicable to protein arrays. Examples are provided from a mantle-cell lymphoma protein-array experiment.
Expression of Human Enhancer of Invasion 10 (HEI10) in Human Myeloid Leukemias and Downregulation during Differentiation of Hematopoietic Stem Cells.
Authors: Fujishima Naohito , Hirokawa Makoto , Fujishima Masumi , Ichikawa Yoshikazu , Horiuchi Takahiro , Miura Ikuo , Sawada Ken-ichi 
Published: April 2020

Abstract
Background: We previously investigated the gene expression profile of drug-resistant human leukemia K562 cells by the serial analysis of gene expression (SAGE) (Ichikawa et al. Int J Hematol, 2004;79:276), and found that the HEI10 gene was overexpressed in K562 cells. With the same technology, we also observed that human CD34+ cells expressed high levels of HEI10 gene expression compared to erythroid progenitor cells (Fujishima et al. Int J Hematol, in press). HEI10 encodes a protein which belongs to a family of E3 ubiquitin ligase,and interacts with cyclin B1 leading to the regulation of cell cycle. In the present study, we examined the gene expression of HEI10 quantitatively in human leukemias in order to learn whether there would be any association between the expression levels and leukemia subtypes. We also investigated whether the expression levels of HEI10 would be related to differentiation of myeloid leukemia cell lines and normal hematopoietic stem cells.
Methods: The quantitative real-time PCR method was employed to estimate the relative gene expression levels of HEI10. Total RNA was extracted from mononuclear cells of bone marrow that had been taken for diagnostic purposes. Patients include acute myeloid leukemia (AML) (n=21) and chronic myeloid leukemia (CML) (n=7; 2 in blastic crisis, 5 in chronic phase). Differentiation of human acute myeloid leukemia cell line HL60 cells was induced by PMA. Human normal CD34+ cells were purified from G-CSF mobilized blood stem cells and were induced to differentiate into erythroid, myeloid or dendritic cells by various cytokines.
Results: Quantitative real-time PCR analysis revealed the high expression of HEI10 gene in patients with leukemia with undifferentiated phenotype (3 cases of AML M1 in FAB classification and 2 cases of CML in blast crisis) and low levels of expression in acute promyelocytic leukemia and CML in chronic phase. When HL60 cells were induced to differentiation with PMA, accumulation in the G0/G1 phase was observed and the expression of HEI10 was decreased. Differentiation of normal CD34+ cells also resulted in a decrease of HEI10 expression.
Conclusions: The expression of HEI10 gene in hematopoietic cells appears to be upregulated when cells are immature. The role for HEI10 in hematopoietic stem cells remains to be elucidated. Biological effects of the HEI10 gene silencing in human hematopoietic cells are currently being investigated by RNAi technology.
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Identification of TAL1/SCL Target Genes through siRNA and Microarray Expression Analysis.
Authors: O’Neil Jennifer , Ferrando Adolfo A. , Palomero Teresa , Look A. Thomas 
Published: April 2020

Abstract
TAL1 is a basic helix-loop-helix (bHLH) protein that is essential for hematopoiesis. Activation of the TAL1/SCL gene occurs in as many as 60% of patients with pediatric and adult T cell acute lymphoblastic leukemia (T-ALL). Mouse models have demonstrated that TAL1 transforms by inhibiting the function of E proteins, however the transcriptional targets of TAL1 that contribute to leukemia are not known. In this study we sought to identify targets of TAL1 that contribute to the transformation of thymocytes through siRNA-mediated knockdown of TAL1 expression in the Jurkat T-ALL cell line that expresses high levels of TAL1. We have generated several clonal cell lines that stably express a siRNA directed against TAL1 with up to 90% knockdown of TAL1 at both the RNA and protein level. Phenotypic characterization of these cells revealed that Jurkat cells with decreased TAL1 expression display a decreased proliferative capacity. This phenotype is augmented in lower serum concentrations in which wild-type and siRNA control Jurkat cells proliferate rapidly. Microarray expression analysis using Affymetrix U133 Plus 2.0 arrays has revealed that TAL1 is a global regulator of gene expression and that many pathways are affected by TAL1 knockdown, including those implicated in cell growth and proliferation. This data will enable us to dissect the downstream pathways of TAL1 by knocking-down or overexpressing these TAL1 targets (depending on whether they are up- or down-regulated in the absence of TAL1), and assessing whether they affect the proliferative rate or survival T-ALL cells. We anticipate that we will identify and validate potential therapeutic targets for TAL1-induced leukemia using this combination of approaches.
Genetic Susceptibility to Hodgkin Lymphoma.
Authors: Cozen Wendy , Salam Muhammad T. , Nieters Alexandra , Ingles Sue A. , Mack Thomas M. , Masood Rizwan , Cockburn Myles G. , Gauderman William J. , Wang Wei , Gill Parkash S. 
Published: April 2020

Abstract
A case-control study of young adult Hodgkin lymphoma (diagnosed before age 50) in twins was conducted to determine if cases and their unaffected identical twins demonstrated dominance of Th1, Th2 or regulatory cytokines compared to unrelated controls. We collected blood specimens from sets of young adult Hodgkin lymphoma cases (n=60), their unaffected identical twins (n=60) and unrelated controls (n=60) and measured IL-6, IL-8, IL-10, and IL-12 levels in PHA-stimulated PBMC supernatant by ELISA. IL-6, IL-10 and IL-12 promoter polymorphisms were determined using PCR and either Taqman or Pyrosequencing. Mean levels of cytokines were compared using an analysis of covariance and genotypes were compared using multivariate logistic regression (SAS). Both cases and their unaffected twins had higher levels of IL-8, but lower levels of IL-12, compared to controls. Unaffected twins of cases had IL-6 levels that were 86% higher than those of controls (p=0.02), however cases’ levels were lower than controls (altered by treatment). Cases and their twins were three times more likely to have an IL-6 genotype associated with higher levels and 2.6 times more likely to have an IL-12 genotype associated with lower levels, compared to controls (Table 2). There were no significant differences in IL-10 levels or genotypes between cases and controls. A genetically determined skewing toward increased inflammatory cytokines (IL-6 and IL-8) and decreased Th1 cytokines (IL-12) is associated with increased susceptibility to young adult Hodgkin lymphoma.
Table 1. Mean and % Difference in Cytokine Levels in Hodgkin Lymphoma Cases, their Unaffected Identical Twins, and Controls
Hodgkin Lymphoma Case v. Control Unaffected Twin of Case v. Control Cytokine Mean Diff (pg/ml) % Diff P-Value Mean Diff (pg/ml) % Diff P-Value IL6 −43.2 −8.7 0.75 +372.8 +83.6 0.04 IL8 +1978.7 +42.9 0.36 +1342.3 +61.9 0.26 IL10 −3.9 −3.0 0.85 −25.6 −20.4 0.20 IL12 −1.2 −54.8 0.01 −0.7 −32.0 0.27
Table 2. Cytokine Genotype and Risk of Young Adult Hodgkin Lymphoma in Cases v. Controls
Cytokine Gene SNP Genotype Odds Ratio 95% Confidence Interval *reference genotype IL6 −174 G&gt;C GG v. CC* 3.12 1.03–9.44 IL12 +1188 A&gt;C CC v. AA* 2.59 1.03–6.55