bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Target Genes of an Erythroid Transcription Factor Complex Containing SCL (TAL1).
Authors: Jim Johanna H. , Dhami Pawandeep , King Amanda , Cooper Jonathan , Vetrie Dave 
Published: October 2019

Abstract
Haematopoiesis is the process whereby haematopoietic stem cells give rise to mature blood cell lineages. The SCL (TAL1) gene, originally identified by chromosomal translocations associated with T-cell acute lymphocytic leukaemia, encodes a key transcription factor (TF) which is expressed in various blood lineages and is essential for haematopoietic development. It has been shown that the SCL protein forms a multi-protein complex during erythroid development with other TFs (GATA1, E2A, LDB1, and LMO2) which binds to a sequence-specific motif to regulate the expression of glycophorin A and c-kit. We have used two complementary approaches to identify novel target genes regulated by this TF complex during erythroid development. In the first approach, we have transfected short interfering RNAs (siRNAs) into the K562 cell line to knockdown transiently the TFs of the erythroid complex. For all members of the complex, a knockdown efficiency of at least 70% was confirmed at the mRNA and protein level within 48 hours after transfection. The consequences of the knockdown at the level of gene expression were observed using Affymetrix GeneChips in order to identify downstream events associated with the erythroid complex in transcriptional programmes. In the second approach, chromatin immunoprecipitation (ChIP) was performed for each member of the complex in the K562 cell line and the ChIP material hybridised to a human transcription factor promoter microarray. A number of novel target genes for the SCL erythroid complex have been identified and verified independently using both approaches. Our data shows that members of the erythroid complex are involved in auto-regulation and regulate genes which control chromatin structure and function. These findings demonstrate that the expression of this TF complex is tightly controlled and point to an important role for it in orchestrating fundamental biological processes which have profound effects on gene expression in erythroid development.
Study on Mechanism of Human Marrow Mesenchymal Stem Cell in Treating Patients with Aplastic Anemia.
Authors: Qiao Zhenhua , Zhao Xiujuan 
Published: October 2019

Abstract
Objective: To explore mechanism of human marrow mesenchymal stem cells (MSCs) in treating patients with aplastic anemia(AA).
Methods:
MSCs in patients with aplastic anemia(AA) and the control group were separated with Percoll(1.073g/m L) and cultured in low glucose DMEM. Then, observed their morphologies,checked their molecule surface antigen by flow cytometry and examined the process of adipogenic differention. The mononuclear cells (MNC)of marrow in patients with AA were enriched based 1.077g/L density centrifuge and cultured in the 1640 medium. (1)MSC in control group and MNC in AA group were co-cultured with or without cytokines. The function of supporting hematopoiesis for MSC was to be observed in single confluence layer after plating by counting the total cells and the clones in every well every week. Then analyzed the dynamics of proliferation. T cells were harvested by using nylon column. MSC in control group and T cells in AA group were co-cultured. The proliferation of T cell was measured by MTT method. The CD25,CD69,CD4,CD8,Annexin-V expression rates of CD3+T cells were analyzed by flow cytometry .The gene and protein of IL-2, IL-4,IL-10,TNF-α,IFN-γ,TGF-β1 were examined by RT-PCR and ELISA respectively. MSC treated to the model of AA, by the examination of peripheral hemogram, bone marrow biopsy, pathological section of spleen.
Results:
There was no significant difference between control group MSC and AA-MSC in morphologies but adipogenic differentiation in AA patients is earlier than controls. The clones of CFU-GM in group(MSC)(78.46±3.58)/2×105 cells, after 14 days cultured was significantly higher than(9.21±4.32)/2×105 cells in group(CK + DMEM medium), while lower than (99.32±4.34)/2×105 cells in group(MSC+CK). (1)the Treg cells (TCD4+CD25+) in AA group (2.01±1.21)/ 2×105 was significantly lower than (4.43±1.67)/2×105 cells in control group, while(5.43±2.31) / 2×105 in group (MSC+AAT) was no more than (4.43±1.67)/2×105 cells in control group. (2) MSCs significantly inhibited T cell proliferation (P< 0. O5)by MTT. (3) RT-PCR and ELISA analysis showed that MSCs induced the expression of IL-4, IL-10, TGF-β1 and decreased significantly the expression of IL-2, TNF-α, IFN -γ in T cells of AA. the model of AA treated by MSCs showed improvements in 3 blood components greatly(p<0.05), Bone marrow proliferated and restored to the normal level, hematopoietic cell increased obviously (hematopoietic cell capacity was more than 40%), and atrophied spleen restore to normality.
Conclusions:
morphologies of AA’MSC had no evident different with the control but was more easy adipogenic differention. aplastic anemia belongs to autoimmune diseases in which T cells effect organ-specific destruction. The fundamental mechanism of MSC in treating AA should be potential to promote hematopoietic cell proliferation by adjusting immunity.
Absence of Oncogenic Transformation in Long-Term Culture Mesenchymal Cells from Bone Marrow of Children.
Authors: Choumerianou Despoina M. , Dimitriou Helen , Perdikogianni Chrysoula , Martimianaki Georgia , Pelagiadis Iordanis , Kalmanti Maria 
Published: October 2019

Abstract
Mesenchymal stromal cells (MSC) have attracted considerable interest in both the scientific and clinical fields because of their potential to regenerate tissues and support hematopoiesis. Although laboratory isolation of MSC is established, the lack of an MSC specific marker and their low frequency in bone marrow necessitates their in vitro expansion in order to obtain a sufficient cell number for applications. During this process their characteristics may be altered and cells may acquire oncogenic properties or abnormal proliferative capacity. We investigated the effect of in vitro expansion on MSC, isolated from BM of children with idiopathic thrombocytopenic purpura (ITP, n=10) and autoimmune neutropenia (AIN, n=6). MSC were maintained in culture for a number of serial passages (P1–P6). We studied the expression profile of tumor suppressor genes (p53, p16, Rb) and the oncogene (H-Ras) along with hTERT, involved in malignant transformation at different passages. The cell line HUVEC was used as normal control for adherent cells and the M5 as positive control for hTERT expression. P2-MSC and P6-MSC (n=9) were grown on soft agar to assess their anchorage dependence. P2-MSC showed normal, relative to GAPDH, expression of p53, p16, Rb and H-Ras (ITP: 0.37, 0.2, 0.53, 0.64 respectively and AIN: 0.41, 0.27, 0.62, 0.75 respectively) compared to HUVEC cells (0.5, 0.23, 0.73, 0.93, respectively) which remained stable upto P6 (ITP P2vsP6: p= 0.63, p= 0.63, p= 0.82, p=0.33, AIN P2vsP6: p= 0.85, p= 0.07, p= 0.14, p=0.07). In all samples, hTERT expression was undetectable both at P2 and P6, and moreover none of the samples showed anchorage independent growth. These findings suggest that expression profile of oncogenes and tumor suppressor genes does not change during in vitro expansion of MSC. Importantly, the similar transcript levels between MSC and HUVEC, the absence of hTERT expression and anchorage independent growth suggest that MSC may not be transformed for a number of passages. In conclusion, although in vivo models are required, such properties encourage MSC use in clinical practice and should be considered as candidates for therapeutic application.
Fog1 and Cebpa Are DNA Targets of GATA-1/PU.1 Antagonism during Leukemia Differentiation.
Authors: Burda Pavel , Curik Nikola , Kokavec Juraj , Pospisil Vit , Skoultchi Arthur I. , Zavadil Jiri , Stopka Tomas 
Published: October 2019

Abstract
PU.1 and GATA1 are hematopoietic lineage-specific transcription factors that play key roles in normal myeloid and erythroid differentiation respectively. Inappropriate expression of PU.1 in proerythroblasts causes its binding to GATA-1 on DNA resulting in a block of erythroid differentiation and development of murine erythroleukemia (MEL) (Rekhtman 1999). Activation of a conditional transgene of GATA-1 (fused with the ligand binding domain of the estrogen receptor, ER) in MEL cells disrupts PU.1-mediated repression in chromatin leading to re-intiation of erythroid differentiation and cell cycle arrest (Choe 2003, Stopka 2005). In this study we show that MEL cells can also be induced to express myeloid differentiation programs upon PU.1-ER activation. Gene expression microarray analysis of GATA-1-ER MEL cells and PU.1-ER MEL cells treated with ER activators allowed us to identify mRNAs that are regulated by both GATA-1 and PU.1 including the set of GATA-1 targets repressed by PU.1, as well as the set of PU.1 targets repressed by GATA-1 in MEL cells. The targets of mutual antagonism of PU.1 and GATA-1 consisted of lineage specific transcription factors, differentiation markers and genes that cause cell cycle arrest and antiapoptotic regulators previously associated with myeloid and erythroid cell differentiation. To determine if PU.1 and GATA-1 directly regulate the lineage specific transcription factor genes, we performed chromatin immunoprecipitation (ChIP) and analyzed the ChIP samples on microarrays and by qPCR. We found that PU.1 and GATA-1 are localized near PU.1 binding sites in the genes for myeloid transcription factors Cebpa and Cbfb and near GATA-1 binding sites in the genes for erythroid transcription factors Fog1 and Nfe2. In addition, further ChIP experiments delineated chromatin architecture near the binding sites for PU.1 and GATA-1 including histone H3 content and acetylation of histone H3K9. We propose that the mutual antagonism of PU.1 and GATA-1 in inhibiting the respective differentiation programs is rendered through specific changes in chromatin structure around lineage specific transcription factors genes. These changes may contribute to the block to differentiation evident in leukemias.
The t(14;18) Is Associated with Germinal Center Phenotype Subset of the Diffuse Large B-Cell Lymphoma Patients in Egypt.
Authors: Abdel-Ghaffar Hasan A. , Abdel-Aziz Sherin M. , Shahin Doaa A. , Board Ezzat S. Sobki , Attwan Nadia I. , Al-Tonbary Youssef A. , Abraham Nader G. 
Published: October 2019

Abstract
Diffuse large B-cell lymphoma (DLBCL) is a generic term for clinically and biologically heterogeneous group of tumors. Identification of high risk patients at presentation will allow effective trials of treatment. Therefore, t(14;18) detection using interphase Florescence in Situ Hybridization (FISH) and Biomed multiplex polymerase chain reaction (PCR) was done on formalin fixed paraffin embedded lymph node archives from pathology department, National Cancer Institute, Cairo, Egypt. Diagnosis were confirmed by pathological review using the diagnostic criteria defined in the revised European-American Classification of Lymphoid Neoplasm / WHO classification. The study was carried out on 26 patients with lymph screen CD 19 +/ CD 5 - / CD 10 ± correlating t(14;18) with the immunophenotypic biological variables, Immunohistochemistry, and the standardized international prognostic index (IPI) with a median follow up for 5 years. Comparison of FISH and PCR techniques showed identical specificity with advantageous sensitivity of FISH over the PCR. Nine patients out of eleven with t(14;18) were associated with Germinal Center (GC) phenotype (CD10+ /Bcl-6 +). However, Only two out of fifteen with non GC phenotype(CD10- /Bcl-6 -) were associated t(14;18). The mean 5 years survival time of patients with t(14;18) was significantly lower (31.18 ± 3.06 month) compared to those without translocation (54.32 ± 2.54 month) (P=0.001). Interestingly, patients with t(14;18) showed Bcl-2 positive (100%) compared to 46.6% in patient without t(14;18) (P=0.004). There is a significant correlation between t(14;18) and the clinicopathological risk criteria of IPI(P=0.01). In our study we demonstrated a detection of t(14;18) by FISH was found to be superior to PCR. The high risk group of GC phenotype together with Bcl-2 expression were associated with t(14;18) and could be used to tailor treatment.
A Study on t(9;22)(q34;q11) in Endothelial Cells in Chronic Myeloid Leukemia.
Authors: Huang Liang , Wu Jingyi , Zheng Miao , Huang Mei , Zhang Yicheng , Zhou Jianfeng 
Published: October 2019

Abstract
Chronic myeloid leukemia (CML) is a disease characterized by t(9;22)(q34;q11), the Philadelphia chromosome (Ph chromosome), and speculated to derive from the hemangioblast, which can give rise to both blood cells and endothelial cells. By using fluorescence in situ hybridization (FISH), we detected Ph chromosome in endothelial cells, which were separated with magnetic beads of CD105+, the marker for endothelial cells, from bone marrow mononuclear cells of 6 CML patients. Moreover, by using Fiction technique, a method combined immunofluorescence with FISH assay, we studied the translocation in vascular endothelium in hepatic and gastroenteric tissue sections and bone marrow trephines of another 7 CML patients. We showed that CD105+ endothelial cells of 6 CML patients with Ph chromosome were identified to harbor the same chromosome aberration as the leukemia cells, 69∼87% of bone marrow endothelial cells carried Ph chromosome. And Ph chromosome was identified in 21∼74% of microvascular endothelial cells in another 7 patients. These results confirmed that CML is derived from the hemangioblast, which can contribute to the maintenance of the blood vascular endothelium. Given specific cytotoxic T lymphocytes can be activated by the immunogenic bcr/abl peptide, we presume that the expression of bcr/abl on endothelial cells in CML patients may be one of the reasons for the incidence of chronic graft versus host disease (cGVHD) after bone marrow transplantation.
FGFR3 Tyrosine Kinase Inhibitor AB1010 as Treatment of t(4;14) Multiple Myeloma.
Authors: Arnulf Bertrand , Ghez David , Leblond Veronique , Choquet Sylvain , Belhadj Karim , Macro Margaret , Bouscary Didier , Facon Thierry , Moreau Philippe , Jaccard Arnaud , Soulier Jean , Karlin Lionel , Hermine Olivier , Fermand Jean-Paul 
Published: October 2019

Abstract
The t(4;14)(p16;q32), found in 15% of multiple myeloma (MM) cases, is associated with a short time to progression (TTP) despite a good initial response. In this subset of MM, relapses are resistant to conventional and intensive chemotherapy leading to a poor prognosis. Plasma cells with t(4;14) ectopically express the fibroblast growth factor receptor 3 (FGFR3), a tyrosine kinase receptor which has proven transforming activity and may represent a therapeutic target. We have studied the safety and efficacy of AB1010, an FGFR3 tyrosine kinase inhibitor, in patients with relapsing/refractory t(4;14) MM. 24 MM patients (M 33%, F 67%, median age 55 years) with t(4;14) were enrolled. FGFR3 expression was detected in all but one patient. AB1010 (9 mk/kg/d) was given orally and Dexamethasone (Dex) (40 mg/d X4d/month) was added if progression. In case of explosive relapse (defined by deep cytopenia, renal failure, circulating plasma cells), a chemotherapy was given followed by a wash out period of 1 month before start of AB1010. Two patients were enrolled after the first line of treatment at a plateau state with a response < 75% (<75% decrease of the monoclonal component). Among the 24 patients, 19 were evaluable since 5 had a duration of treatment < 1month. The main toxicities were gastrointestinal (nausea 63,5%, grade I/II, diarrhoea 25%, grade I; anorexia 25%, grade II) and oedema (face 50%, grade I/II and legs 24% grade I). Hematological toxicities were limited to a transient grade III neutropenia in one patient. Dex was added in all cases. Six patients (3 in 1st relapse, 3 in >3rd lines) had explosive relapse. Treatment regimen before start of AB1010 were :VTD (4); Thal/Dex (1); MLPHD (1). In these 6 cases, the TTP range from 1,5 to 4 months. 11 patients had non explosive relapse. Of these, 5 were in 1st relapse in whom 1 near CR (figure) and 1 PR were observed, with a median TTP of 10 months. Six patiens were in > 2nd relapse and 2 MR were obtained. Of the 2 remaining patients enrolled at plateau, 1 progressed at 6 and 1 is still on going after 13 months. In conclusion, AB1010, in combination with Dex, is well tolerated and may be useful in the treatment of patients with t(4;14) MM, especially in early phase of the disease. Responses were observed (40%) in patients with first relapse with a TTP superior as compared to the reported one (4,7 months with Thalidomide/Dex, Jaksic et al, 2006). Given the synergistic effect on MM cells proliferation observed in vitro, a study is now on going to test the safety and the efficacy of the combination AB1010 + Dex + Velcade in early relapsing patients with t(4;14) MM.
Figure Figure
SMRT; Not So Smart in Multiple Myeloma.
Authors: Ghoshal Pushpankur , Houde Christiane , Szafranek Angela , Nganga Alain , Johnson Timothy , Bigelow Ashley , Moran-Guiati Joseph , Smiraglia Dominic , Chanan-Khan Asher Alban , Coignet Lionel J. 
Published: October 2019

Abstract
Multiple myeloma (MM), a clonal B-cell malignancy, characterized by accumulation of plasma cells in bone marrow is the second most hematological malignancy in United States. In the BM, the myeloma and stromal cells secrete cytokines, which support the growth and survival of the myeloma cells. We previously showed that JAG2, one of the NOTCH ligand, is over-expressed in MM cells (Blood. 2004 Dec 1;104(12)). We hypothesized that over-expression of JAG2 in myeloma cells induce secretion of IL-6 from the stromal cells and subsequently enhances the proliferation of myeloma cells. JAG2 has been shown to be over-expressed in all cell lines and patient samples studied To identify the mechanism for JAG2 overexpression in MM cells, we assessed the potential modifications of the JAG2 promoter in MM cell lines as well as patient samples (an JAG2 negative controls) by studying both methylation and histone acetylation level of the JAG2 promoter. We show that difference in H4 acetylation level might play a crucial role in JAG2 expression. Acetylation state of histones can be regulated by the recruitment of histone deacetylases (HDACs). HDACs are typically recruited to promoter regions through interaction with nuclear co-repressors such as SMRT. The cell lines and patient samples studied presented significantly reduced levels of SMRT. Therefore, based on these observations we propose a model in which partial down-regulation of SMRT recruits less active HDAC3 (as confirmed by immunoprecipitation). As a result the deactylation process of histones in the JAG2 promoter region has been impaired and the cells lost their regulatory mechanism on transcriptional regulation of JAG2. This provides a mechanistic explanation for JAG2 over-expression in MM and on the direct involvement of the SMRT co-repressor in MM pathogenesis.
Insertional Activation of GLI2 in Adult T-Cell Leukemia/Lymphoma.
Authors: Cleveland Susan M. , Dave Utpal P. 
Published: October 2019

Abstract
Retroviruses induce cancer by integrating into the cellular genome and activating oncogenes or inactivating tumor suppressor genes. Human T-cell Leukemia Virus type 1 (HTLV-1), a complex retrovirus, induces Adult T-cell Leukemia/Lymphoma (ATLL) after a latency of over 30 years and in only 5% of carriers. The long latency and incomplete penetrance is similar to how slow transforming retroviruses induce cancer in mice and imply multiple oncogenic “hits” need to accumulate for clinically apparent disease. Insertional mutagenesis may be one mechanism by which ATLL develops. We used splinkerette-PCR to clone and map insertion sites from an HTLV-1 infected T-cell line, Hut-102. We identified an HTLV-1 insertion 5′ of the GLI2 gene, formerly known as Tax-Helper-Protein-1. We found GLI2 was up-regulated by promoter insertion. Interestingly, we found GLI2 protein occupied the HTLV-1 Long Terminal Repeat. The effect of GLI2 expression on viral expression was investigated by knockdown of GLI2 in Hut-102 cells. Our results show that retroviral insertional mutagenesis can be an important mechanism in HTLV-1-induced leukemias and lymphomas.
Mechanism of Tetrandrine in Combination with Droloxifen To Reverse Multidrug Resistance in Leukemia.
Authors: Chen Bao-An , Su Ai-ling , Gao Feng , Cheng Jian , Ding Jia-Hua , Gao Chong , Sun Yun-yu , Wang Jun , Zhao Gang , Chen Ning-Na , Zhao Hui-Hui , Pei Xiao-Ping 
Published: October 2019

Objective: The study aims to investigate the effect of Tetrandrine (Tetrandrine, Tet) in combination with Droloxifen (Droloxifen, DRL) on the expression of NF-κB(nuclear factor kappaB) of K562 and K562/A02 cell lines and the reversal mechanism of this combination.
Method:
The activation of NF-κB in K562 and K562/A02 cell lines and the effect of Tet,DRL alone or in combination on NF-κB were determined with Immunocytochemistry and Western blotting respectively.
Results:
K562/A02 cells displayed higher level of NF-κB protein expression than their parental K562 cells; The application of Tet or DRL alone or in combination had no effect on NF-κB expression in K562 cells at 6h and 12h; Tet and DRL used alone or in combination could significantly down-regulate NF-κB protein expression in nucleaus of K562/A02 cells.
The effect was more significant in combination of Tet and DRL than the application of single drug. This decrease became more significant at 12h than at 6h.
Conclusions:
Activation of NF-κB may be involved in the mechanism of MDR of K562/A02 cell line; Inhibition of NF-κB activation may be involved in the mechanism of multidrug resistance reversal to K562/A02 line of Tet and DRL.