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Effect of loss of Gfi1b on hematopietic stem cells

ABSTRACT: Using Gfi1b conditional mice, deletion of gfi1b in the hematopietic system was induced by injecting MxCre tg Gfi1bfl/fl mice with pIpC. 30 days after injection, Cd150 pos, Cd 48 neg, Lin neg Sca and c-kit pos stem cells were sortrted from Gfi1bfl/fl and Mxcre tg Gfi1bfl/fl mice and analysed. We used the mouse Affymetrix Gene ST Array.

ORGANISM(S): Mus musculus

PROVIDER: GSE20655 | GEO | 2010/09/03

SECONDARY ACCESSION(S): PRJNA124877

REPOSITORIES: GEO

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Effect of loss of Gfi1b on hematopietic stem cells

Project description:Using Gfi1b conditional mice, deletion of gfi1b in the hematopietic system was induced by injecting MxCre tg Gfi1bfl/fl mice with pIpC. 30 days after injection, Cd150 pos, Cd 48 neg, Lin neg Sca and c-kit pos stem cells were sortrted from Gfi1bfl/fl and Mxcre tg Gfi1bfl/fl mice and analysed. We used the mouse Affymetrix Gene ST Array. The study should determine whether loss of Gfi1 alters the gene expression pattern in the hematopietic stem cells.

2010-09-03 | E-GEOD-20655 | biostudies-arrayexpress

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Growth factor independence 1b (Gfi1b) is required for erythroid cell maturation and regulates embryonic globin expression [Mouse430_2]

Project description:Growth factor independence 1b (Gfi1b) is a DNA binding repressor of transcription with vital functions in hematopoiesis. Gfi1b-null embryos die at midgestation very likely due to defects in erythro- and megakaryopoiesis. To analyze the full functionality of Gfi1b in erythropoiesis, we used conditionally deficient mice that harbor floxed Gfi1b alleles and the Mx-Cre transgene inducible by pIpC treatment. We used bone marow cells from 12 weeks old wild type C57BL/6 mice or conditional Gfi1b knock-out mice, both carrying the Mx-cre transgene induced by five treatments with pIpC (500µg each) every other day and sacrificed 42 days after the first injection. Ter119 positive erythroid cells were isolated by flow cytometry and RNA from these cells was isolated using the Trizol procedure according to the manufacturers instructions for gene expression analysis on Affymetrix MOE_430 2.0 expression arrays (GPL 1261)

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Comparative transcriptomics reveals circadian and pluripotency networks as two pillars of longevity regulation

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Expression data for normal flow sorted hematopietic cell subpopulations

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RNA-Seq Analysis of Gfi1b KO Hematopoietic Stem Cells

Project description:The zinc finger protein and SNAG domain transcription factor Gfi1b is highly expressed in hematopoietic stem cells (HSC) and in megakaryocytes (MKs). Deletion of Gfi1b in mice leads to a drastic expansion of both cell types, suggesting that Gfi1b controls their proliferation. Here we present evidence that Gfi1b exerts this control by modulating the Wnt/beta-catenin signaling pathway. We can show that Gfi1b binds to beta-catenin and is part of a larger complex that contains β-catenin co-factors. Gfi1b activates beta-catenin/TCF mediated transcription and is required for the activity of beta-catenin target gene promoter driven reporter genes in vivo. This is dependent of the ability of Gfi1b to recruit the histone modifying enzyme lysine demethylase 1 (LSD1) to β-catenin containing complexes. Moreover, LSD1 enhances the Gfi1b-mediated activation of beta-catenin/TCF dependent transcription. Both Gfi1b deficient HSCs and MKs show de-regulation of expression of many sets on Wnt/β-catenin target genes and Gfi1b and β-catenin co-occupy the promoters of many common target genes. Finally, activating the Wnt/β-catenin signaling pathway in Gfi1b deficient HSCs and MKs significantly reduces their expansion, which confirms that Gfi1b is a critical factor controlling the cellularity of HSCs and MKs and exerts this function by regulating the Wnt/β-catenin pathway in these cells.

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GFI1B and LSD1 repress myeloid traits during megakaryocyte differentiation

Project description:The transcription factor Growth Factor Independence 1B (GFI1B) recruits Lysine Specific Demethylase 1A (LSD1/KDM1A) to stimulate gene programs relevant for megakaryocyte and platelet biology. Inherited pathogenic GFI1B variants result in thrombocytopenia and bleeding propensities with varying intensity. Whether these affect similar gene programs is unknow. Here we studied transcriptomic effects of four patient-derived GFI1B variants (GFI1BT174N,H181Y,R184P,Q287*) in MEG01 megakaryoblasts. Compared to normal GFI1B, each variant affected different gene programs with GFI1BQ287* uniquely failing to repress myeloid traits. In line with this, single cell RNA-sequencing of induced pluripotent stem cell (iPSC)-derived megakaryocytes revealed a 4.5-fold decrease in the megakaryocyte/myeloid cell ratio in GFI1BQ287* versus normal conditions. Inhibiting the GFI1B-LSD1 interaction with small molecule GSK-LSD1 resulted in activation of myeloid genes in normal iPSC-derived megakaryocytes similar as observed for GFI1BQ287* iPSC-derived megakaryocytes. Thus, GFI1B and LSD1 facilitate gene programs relevant for megakaryopoiesis while simultaneously repressing programs that induce myeloid differentiation.

2024-03-28 | PXD050401 | Pride

RNA-Seq Analysis of Gfi1b KO Megakaryocytes

Project description:Megakaryocytes isolated from Gfi1b flox/flox mice carrying PF4-Cre or not, and from Gfi1b flox/flox mice carrying ROSA-Cre-ERT with or without tamoxifen injection were analyzed for differential expression by RNA-Seq

2017-01-21 | GSE71310 | GEO

Overexpression of Gfi1b in a pro-B Abelson murine leukemia virus transformed cell line

Project description:We have conducted a screen for factors that downregulate expression of the genes encoding the V(D)J recombinase (RAG1 and RAG2) during B cell development. We have identified the transcription factor Gfi1B as being one of the proteins capable of decreasing RAG transcription when overexpressed in Ableson transormed ProB cell lines. We have yet to determine whether the overexpression of Gfi1B downregulates the RAGs directly, or whether it initiates a signalling programme that results in RAG downregulation. We hypothesize that by comparing global gene expression patterns in cells that overexpress Gfi1B and those that do not, we can distinguish between these possibilities and additionally gain insight into the broader genetic program that may be influenced by Gfi1B during hematopoiesis. Abelson pro-B cells were infected with a retrovirus encoding Gfi1b fused to the estrogen receptor domain. Gfi1b expression was induced by adding tamoxifen to the culture medium for 12h. Three biological replicates of untreated and treated cells were analyzed.

2011-12-13 | E-GEOD-33709 | biostudies-arrayexpress

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