Project description:Pediatric severe aplastic anemia (PSAA) is a rare and life-threatening disease, which has been suggested to result from deficiency in essential cytokines or growth factors in bone marrow mesenchymal stem cells (MSCs). In this study, we examined expression profiles of the essential cytokines and growth factors, which were involved in proliferation and differentiation of MSC, in 5 PSAA patients using oligonucleotides microarrays. Following evaluation of gene expression patterns using cluster analysis and gene ontology classifications, fifteen potential genes that were cell proliferation- and cytokine-related and significantly down-regulated in PSAA were selected. The findings of the microarray analysis were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Interestingly, silencing of chemokine ligand 12 (CXCL12, also named stromal cell-derived factor 1, SDF1) or hepatocyte growth factor (HGF) gene expression reduced cell growth and response to adipogenic factor-induced differentiation, but increased osteogenesis, which possibly derived from hematopoietic stem cells (HSCs). Re-introduction of the CXCL12 gene or exogenously added HGF, on the other hand, partially recovered cell function of PSAA MSC. These results suggest that gene expression in PSAA MSC is globally down-regulated by a yet-to-be-determined mechanism. Among these down-regulated genes, CXCL12 and HGF played vital roles in regulating cell growth and differentiation. Mesenchymal stem cells from five pediatric SAA patients vs. MSC from five healthy donors, which were pooled together.
Project description:Bulk RNA-seq data of Lin-CD34+ hematopoietic stem and progenitor cells derived from bone marrow of healthy donors and untreated aplastic anemia patients
Project description:We performed single cell RNA sequencing, and VDJ sequencing of TCR and BCR of bone marrow samples (BMMMNCs and sorted CD34+ HSPCs) from 20 patients with severe aplastic anemia (SAA), pre- and post-treatment, to understand disease pathogenesis and response to treatment.
Project description:Transcriptome analysis of hematopoietic stem and progenitor cells (HSPCs) and T cells collected from bone marrow and peripheral blood of healthy donors and aplastic anemia patients untreated or response to immunosuppressive therapy.
Project description:Bone marrow nucleated cells (BMNCs) from healthy donors and patients with non-severe aplastic anemia (NSAA) at the time of initial diagnosis were separated, and then co-cultured with or without 40 μg/mL levamisole (LMS) for 48 hours in vitro. Cells were harvested and total RNA were extracted. The sequencing assay were carried out using the Illumina HiSeq X platform and the data obtained were analyzed by following the Hisat2 protocol.
Project description:CD3+ T cells derived from bone marrow aspirates and peripheral blood samples of newly diagnosed AA patients and healthy volunteers were analysed using Affymetrix HG_U133A GeneChips. Additionally, two patients were studied after achieving a partial remission (post-Therapy). Experiment Overall Design: Altogether, we analysed 8 different samples. Pool I contained three female and one male patients, 19 to 70 years old, suffering from (severe) aplastic anemia. From these four donors, samples derived from bone marrow aspirates and from peripheral blood, were pooled on total-RNA basis. Pool II consisted of additional two patients, one male (64 years old) and one female (70 years old). From both donors, total-RNA was extracted and pooled from CD3+ T cells derived either from bone marrow or peripheral blood, each prior and post therapy. Healthy control pools included three donors for the peripheral blood sample and two donors for bone marrow derived T cells.
Project description:Fanconi anemia (FA) is a rare inherited disease complicated by aplastic anemia. There is evidence that hematopoietic stem cells have lost self replicative capacity and undergo apoptosis when exposed to inhibitory cytokines including interferon gamma and tumor necrosis factor-alpha. We used gene expression microarrays to identify transcriptomal differences between bone marrow cells from normal volunteers and from children and adults with Fanconi anemia Experiment Overall Design: Fanconi anemia patients were identified using mitomycin C and/or diepoxybutane chromosomal breakage analysis. Eleven normal volunteers and 21 FA patients were studied. All FA patients with cytogenetic evidence of clonal evolution were excluded. All FA patients with acute leukemia were excluded. RNA was prepared from freshly obtained low density mononuclear cell fractions.
Project description:Fanconi anemia (FA) is a rare inherited disease complicated by aplastic anemia. There is evidence that hematopoietic stem cells have lost self replicative capacity and undergo apoptosis when exposed to inhibitory cytokines including interferon gamma and tumor necrosis factor-alpha. We used gene expression microarrays to identify transcriptomal differences between bone marrow cells from normal volunteers and from children and adults with Fanconi anemia
Project description:During immune-mediated severe aplastic anemia (SAA) monocytes (CD11b+ Ly6C+ Ly6G-) signficantly increase by both frequency and number within the bone marrow. We isolated BM monocytes from F1 hybrid mice (C57BL/6;Balb/c) induced with SAA via the splenocyte-transfer model. We utilized single-cell RNA sequencing (scRNA-seq) to analyze the heterogenity of BM monocytes during SAA.
Project description:Pediatric severe aplastic anemia (PSAA) is a rare and life-threatening disease, which has been suggested to result from deficiency in essential cytokines or growth factors in bone marrow mesenchymal stem cells (MSCs). In this study, we examined expression profiles of the essential cytokines and growth factors, which were involved in proliferation and differentiation of MSC, in 5 PSAA patients using oligonucleotides microarrays. Following evaluation of gene expression patterns using cluster analysis and gene ontology classifications, fifteen potential genes that were cell proliferation- and cytokine-related and significantly down-regulated in PSAA were selected. The findings of the microarray analysis were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Interestingly, silencing of chemokine ligand 12 (CXCL12, also named stromal cell-derived factor 1, SDF1) or hepatocyte growth factor (HGF) gene expression reduced cell growth and response to adipogenic factor-induced differentiation, but increased osteogenesis, which possibly derived from hematopoietic stem cells (HSCs). Re-introduction of the CXCL12 gene or exogenously added HGF, on the other hand, partially recovered cell function of PSAA MSC. These results suggest that gene expression in PSAA MSC is globally down-regulated by a yet-to-be-determined mechanism. Among these down-regulated genes, CXCL12 and HGF played vital roles in regulating cell growth and differentiation.
