JAK2V617F mutant HSC microarray
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Anthony Green
PROVIDER: E-MTAB-6878 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:JAK2V617F mutation is found in most patients with a myeloproliferative neoplasm (MPN), including polycythemia vera (PV), essential thrombocythemia (ET). We have demonstrated that heterozygous human JAK2V617F are associated with ET-like phenotypes while homozygosity for human JAK2V617F results in a striking phenotypic switch from an ET-like to PV-like phenotype. The resultant erythrocytosis is driven by increased numbers of erythroid progenitors and enhanced erythroblast proliferation. To establish the molecular mechanisms and pathways involved in the erythrocytosis, microarray was performed on bone marrow erythroblasts isolated from 8-10 weeks old homozygous, heterozygous mice and wildtype controls (3 mice for each genotype).
2014-03-26 | E-MTAB-2427 | biostudies-arrayexpress
Project description:Comparison of mRNA expression profiles of LT-HSCs with or without mutations in JAK2 and Ezh2 by RNA sequencing. LT-HSC mRNA was extracted from six different transgenic mice (SclCre, SclCre;Ezh2+/-, SclCre;Ezh2-/-, SclCre; JAK2V617F, SclCre; JAK2V617F;Ezh2+/-, SclCre; JAK2V617F;Ezh2-/-) 10 weeks after tamoxifen injection. Our study represents the first detailed analysis of mRNA expression profile of LT-HSC with or without mutations in JAK2 and Ezh2 , with biologic replicates, generated by RNA-seq technology. Our results revealed that mRNA expression profile of LT-HSC with different genotype showed specific gene expression patterns, which allows to do biological comprehensive and quantitative analysis for hematopoiesis. LT-HSCs mRNA profiles six different transgenic mice (SclCre, SclCre;Ezh2+/-, SclCre;Ezh2-/-, SclCre; JAK2V617F, SclCre; JAK2V617F;Ezh2+/-, SclCre; JAK2V617F;Ezh2-/-) were generated by deep sequencing.
2016-06-24 | E-GEOD-82073 | biostudies-arrayexpress
Project description:Objective: Microarray analysis was used to determine the molecular mechanism underlying Fancd2 and Foxo3a double knockout mice HSCs exhaustion. Methods: SLAM cells were obtained from WT, Fancd2KO, Foxo3aKO and DKO bone marrow cells. Total RNA from SLAM cells was purified by RNeasy kit (Qiagen) following the manufacturer’s procedure. And then the RNA was fluorescently labeled and hybridized to Affmetrix Mogene 2.0 ST array. Data was analyzed by Genespring GX11 software. Results: a large proportion of differentially expressed genes in dKO HSCs belonged to cell cycle control. Another group of up-regulated genes were those involved in the differentiation of HSCs. Other differentially regulated transcripts included genes known to be involved in oxidative stress response, or to have roles in DNA repair. Conclusion: Cell cycle, DNA repair, DNA binding and hematopoietic lineage differentiation pathways might have been perturbed in HSC exhaustion. Mouse Femurs and tibias from WT, Fancd2KO, Foxo3aKO and DKO mice were flushed to dissociate the BM fraction. The mononuclear cells were isolated by Ficoll (GE Healthcare) gradient centrifugation. Single SLAM (Lin-ckit+Sca-1+CD34-) cells were sorted. RNA was extracted, labelled, and quantified using the Mouse ST-2 microarray.
2014-12-17 | E-GEOD-64215 | biostudies-arrayexpress
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2016-05-19 | E-GEOD-67120 | biostudies-arrayexpress
Project description:Hematopoietic stem cells (HSCs) in adult are specified early from the endothelium-derived precursors (e.g., hemogenic endothelium, and pre-HSCs) in mouse mid-gestation embryos, the detailed process, however, is still largely unknown due to their rareness, transience, and current inability to prospectively isolate them efficiently . Here we developed a potent set of surface markers that could capture the earliest emerging HSCs, the CD45- pre-HSCs with high accuracy and purity, as rigorously and functionally verified by single-cell-initiated serial transplantation assays. Then we applied single-cell RNA-Seq technique to analyze five populations related to HSC formation: the CD45- (type 1) and CD45+ (type 2) pre-HSCs as well as endothelial cells in the E11 AGM region; and later mature HSCs in the E12 and E14 fetal livers. Compared to other cell populations, both type 1 and type 2 pre-HSCs have their unique signatures of transcription machinery, transcription factor network, signaling pathway, cell cycle status, metabolism state, and lncRNA expression patterns. Our work paves the way for dissection of the complex molecular mechanisms regulating the step-wise formation of HSCs from endothelial cells, thus informing future efforts on engineering HSCs for clinical application. RNA-Seq of 35 10-cell pooled samples from 5 FACS sorted cell types, i.e., endothelial cells (ECs), T1 and T2 pre-HSCs from E11 AGM region, as well as mature HSCs from E12 and E14 fetal liver
2016-05-19 | E-GEOD-66954 | biostudies-arrayexpress
Project description:MTD project_description Inflammation and decreased stem cell function characterize organism aging, yet the relationship between these factors remains incompletely understood. This study shows that aged hematopoietic stem and progenitor cells exhibit increased ground-stage NF-κB activity, which enhances their responsiveness to undergo differentiation and loss of self-renewal in response to inflammation. The study identifies Rad21/cohesin as a critical mediator of NF-κB signals, by increasing chromatin accessibility of inter-/intra-genic and enhancer regions. Rad21/NF-κB are required for normal differentiation, but limit self-renewal of hematopoietic stem cells (HSCs) during aging and inflammation in an NF-κB dependent manner. HSCs from aged mice fail to downregulate Rad21/cohesin and inflammation/differentiation inducing signals in the resolution phase after acute inflammation. and The inhibition of cohesin/NF-κB is sufficient to revert the hypersensitivity of aged HSPCs to inflammation-induced differentiation. During aging, myeloid-biased HSCs with disrupted and naturally occurring reduced expression of Rad21/cohesin are increasingly selected over lymphoid-biased HSCs. Together, Rad21/cohesin mediated NF-κB signaling limits HSPC function during aging and selects for cohesin deficient HSCs with myeloid skewed differentiation.
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Project description:To determine genes in FL HSCs that are sensitive to Notch signagling, E14.5 FL cells were cultured on DL1( to stimulate Notch signaling). Cells were cultured in the presence of DMSO (vehicle control) or gamma secretase inhibitor (1uM) for 4 hrs or 10hrs. Gamma secretase inhibitor was used to inhibit Notch signaling. SLAM-LSKs were sorted and used for RNA preparation. 7 biological replicates (14 totalt samples) were collected for cells cultured for 4hrs. 4 biological replicates (8 total samples) were collected from cells cultured for 10hrs. Each replicate consisted of a DMSO and GSI treated sample
2014-01-28 | E-GEOD-53713 | biostudies-arrayexpress
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Study Design: Single arm Non-randomized
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