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Expression data from G0 versus G4/G5 CMP/GMP

ABSTRACT: Telomere dysfunctional CMP/GMP have deregulated pathways that are associated with DNA damage signaling We compared differentially expressed genes in the G4/G5 CMP relative to the G0 control to identify pathways that may affect CMP differentiation.

ORGANISM(S): Mus musculus

PROVIDER: GSE62278 | GEO | 2015/04/09

SECONDARY ACCESSION(S): PRJNA263712

REPOSITORIES: GEO

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Expression data from G0 versus G4/G5 CMP/GMP

Project description:Telomere dysfunctional CMP/GMP have deregulated pathways that are associated with DNA damage signaling We compared differentially expressed genes in the G4/G5 CMP relative to the G0 control to identify pathways that may affect CMP differentiation. Bone marrow CMP and GMP cells were sorted from two paired pools of G0 TERTER/+ or G4/G5 TERTER/ER mice (5,000-20,000 cells per sample) using the Influx Cell Sorter. Every paired pool includes CMP or GMP sorted from 4 age and gender matched G0 or G4/G5 mice. RNA from the respective sorted cells was extracted using Trizol (Ambion) and profiled on 2100 Bioanalyzer (Agilent). Gene expression profiling was performed at the Sequencing and Non-coding RNA Program at MD Anderson Cancer Center. Briefly, the GeneChip® 3 IVT Express Kit (Affymetrix) was used to generate biotin-labeled cRNA, which were purified and fragmented, before target hybridization on the GeneChip® Mouse Genome 430 2.0 Array (Affymetrix) according to the manufacturer's instructions. Affymetrix raw data (CEL files) were normalized using Affymetrix Microarray Suite (MAS) version 5.0 using a TGT=100. Paired pools used in the study were: CMP pool 1: G0-1 and G4/5-1 CMP pool 2: G0-2 and G4/G5-2 GMP pool 1: G0-1 and G4/G5-1 GMP pool 2: G0-2 and G4/G5-2

2015-04-09 | E-GEOD-62278 | biostudies-arrayexpress

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2015-04-09 | GSE62392 | GEO

Reduced representation bisulfite sequencing from G0 & G4/G5 CMP

Project description:Differential hyper- and hypo-methylation regions in G0 versus G4/G5 CMP The goal of this study is to evaluate changes in CpG methylation profilings of telomere dysfunctional common myeloid progenitor cells (CMP) as compared to their wild type controls Genomic DNA was extracted from sorted CMP populations isolated from 3 pools of G0 or 2 pools of G5 mice using UltraPure Phenol:Chloroform:Isoamyl Alcohol according to manufacturer’s instructions (Life Technologies). 14,000 to 30,000 cells were available for each sample, resulting in a minimum of 45ng of DNA. Genome-wide DNA methylation profiling was performed by RRBS. Library preparation and sequencing were performed at the UT MD Anderson Cancer Center’s DNA Methylation Analysis Core and Sequencing and Microarray Facility, according to published protocols. RRBS sequencing data were aligned and methylation was called using Bismark v0.7.119. In brief, bisulphite-treated DNA was aligned to UCSC Genome Browser mm10 reference genome using Bowtie. In total 29-38 million reads were generated per sample with alignment rates around 63%. Next, MethylKit10 implemented with Fisher’s exact test was used to compare the cytosine methylation profiles of G0 and G5 CMP. Gene promoter regions were calculated based on RefSeq gene annotations with regions starting 1 kb upstream of the annotated transcription start site (TSS) and extending 500 base pairs downstream of TSS. Exons, introns, and CpG islands coordinates were collected from the UCSC Genome Browser mm10 version.

2015-04-09 | E-GEOD-62392 | biostudies-arrayexpress

Mus musculus

Project description:Expression data from G0 versus G4/G5 CMP/GMP

| PRJNA263712 | ENA

Mus musculus

Project description:Reduced representation bisulfite sequencing from G0 & G4/G5 CMP

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Transcription profiles of WT and CreER-Raptor-flox/flox common myeloid progenitors (CMP), granulocyte-macrophage progenitor (GMP), or lineage negative (Lin-) stimulated with M-CSF for 12 hours

Project description:Myelopoiesis is impaired in Raptor-deleted mice (CreER-Rptor-flox/flox). To evaluate the transcriptional changes in myeloid precursors , we isolated CMP (Lin–Sca-1–c-Kit+CD34+FcγRII/IIImid), GMP (Lin–Sca-1–c-Kit+CD34+FcγRII/IIIhigh) and Lin (B220, Ly6C, Ly6G, CD3, Ter-119) negative cells (Lin–) from bone marrow of WT or CreER-Rptor-flox/flox mice. RNA was isolated from CMP and GMP immediately after sorting and Lin- cells were cultured for 12 hours with M-CSF (10 ng/mL) in 10% FBS and 1% P/S DMEM before RNA isolation. We used microarrays to compare the global transcription profiles of CMP, GMP, or Lin– stimulated with M-CSF for 12 hours from WT and CreER-Raptor-flox/flox mice.

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Transcription profiling of mouse hematopoietic cells (GMP, CMP, CLP and HSC), FACS sorted from wild type and Mll-AF9 knock-ins

Project description:The pathways by which oncogenes, such as MLL-AF9, initiate transformation and leukemia in humans and mice are incompletely defined. In a study of target cells and oncogene dosage, we found that Mll-AF9, when under endogenous regulatory control, efficiently transformed LSK (Lin- Sca1+ c-kit+) stem cells while committed granulocyte-monocyte progenitors (GMPs) were transformation-resistant and did not cause leukemia. Mll-AF9 was expressed at higher levels in hematopoietic stem (HSC) than GMP cells. Mll- AF9 gene dosage effects were directly shown in experiments where GMPs were efficiently transformed by the high dosage of Mll-AF9 resulting from retroviral transduction. Mll-AF9 up-regulated expression of 196 genes in both LSK and progenitor cells, but to higher levels in LSKs than in committed myeloid progenitors. Experiment Overall Design: Comparison of gene expression profiles among four types of hematopoietic cells (GMP, CMP, CLP and HSC), FACS sorted from wild type and Mll-AF9 knock-in mice. The goal was to identify genes differentially expressed in each Mll-AF9 cell type compared to the corresponding wild type cells.

2008-04-07 | E-GEOD-10627 | biostudies-arrayexpress

Genome-wide occupancy of H3K27me3 in human gingival mesenchymal and periodontal ligament stem cells in response to Electronic-cigarette

Project description:The epigenetic regulation on gene transcription affected by Electronic-cigarette (E-cig) to human gingival mesenchymal (GMSC) is not fully understood. Here, we collected GMSCs from systemically healthy patients due to the premolars and extracted for orthodontic reasons, and treated with E-cig smoke with original flavor (G2), E-cig smoke with menthol flavor (G3), E-cig liquid with original flavor (G4), and E-cig liquid with menthol flavor (G5) by the optimal conditions of LC50, and conducted H3K27me3 ChIP-seq to compare with untreated control (G0).

2022-09-03 | E-MTAB-12079 | biostudies-arrayexpress

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Transcriptome characterization of human gingival mesenchymal and periodontal ligament stem cells in response to Electronic-cigarette

Project description:The change of transcriptome and related biological functions affected by tobacco smoke and Electronic-cigarette (E-cig) to human gingival mesenchymal (GMSC) and periodontal ligament stem cells (PDLSCs) are not fully understood. Here, we collected GMSCs and PDLSCs from systemically healthy patients due to the premolars and extracted for orthodontic reasons, and treated with tobacco smoke (G1, P1), E-cig smoke with original flavor (G2, P2), E-cig smoke with menthol flavor (G3, P3), E-cig liquid with original flavor (G4, P4), and E-cig liquid with menthol flavor (G5, P5) by the optimal conditions of LC50, and conducted RNA-seq to compare with untreated control (G0, P0).

2022-06-17 | E-MTAB-11797 | biostudies-arrayexpress

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