Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of Mac-1+, c-Kit+ myeloid progenitor cells from mice homozygous for the p42-specific knock-out Cebpa mutation and independently transplanted with leukemic bone marrow, and from control mice heterozygous for Cebpa mutation

ABSTRACT: 5000 Mac-1+, c-Kit+ myeloid progenitor cells from 3 mice homozygous for the p42-specific knockout Cebpa mutation and independently transplanted with leukemic bone marrow, or from 3 control mice heterozygous for Cebpa mutation, were isolated by FACS. Total RNA was extracted and fluorescence labeled for hybridization to MOE430.2 gene expression arrays. cRNA synthesis entailed 2 rounds of linear amplification using the T7 cRNA protocol provided by the manufacturer. Hybridization and washing was performed according to Affymetrix GeneChip expression analysis technical protocols. Microarray data were analyzed in the R statistical computing framework using the Bioconductor suite. Hybridization quality was assessed with the Bioconductor package arrayQualityMetrics and the raw fluorescence data were processed using the GCRMA method.

ORGANISM(S): Mus musculus

SUBMITTER: Claus Nerlov

PROVIDER: E-MEXP-1444 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Modeling of C/EBPalpha mutant acute myeloid leukemia reveals a common expression signature of committed myeloid leukemia-initiating cells.

Kirstetter Peggy P Schuster Mikkel B MB Bereshchenko Oksana O Moore Susan S Dvinge Heidi H Kurz Elke E Theilgaard-Mönch Kim K Månsson Robert R Pedersen Thomas A TA Pabst Thomas T Schrock Evelin E Porse Bo T BT Jacobsen Sten Eirik W SE Bertone Paul P Tenen Daniel G DG Nerlov Claus C

Cancer cell 20080401 4

Mutations in the CEBPA gene are present in 7%-10% of human patients with acute myeloid leukemia (AML). However, no genetic models exist that demonstrate their etiological relevance. To mimic the most common mutations affecting CEBPA-that is, those leading to loss of the 42 kDa C/EBPalpha isoform (p42) while retaining the 30kDa isoform (p30)-we modified the mouse Cebpa locus to express only p30. p30 supported the formation of granulocyte-macrophage progenitors. However, p42 was required for contr ...[more]

PMID: 18394553

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Project description:C/EBPalpha is a transcription factor critically involved in myeloid development and indispensable for formation of granulocytes. To track the cellular fate of stem and progenitor (LSK) cells, which express C/EBPalpha, we developed a mouse model expressing Cre recombinase from the Cebpa promoter and an inducible EYFP allele. We show that Cebpa/EYFP+ cells represent a significant subset of LSK cells, which predominantly give rise to myeloid cells in steady state hematopoiesis. C/EBPalpha induced a robust myeloid gene expression signature and downregulated E2A-induced regulators of early lymphoid development. In addition, Cebpa/EYFP+ cells comprise a fraction of early thymic progenitors (ETP) with robust myeloid potential. However, Cebpa/EYFP+ LSK and ETP cells retained the ability to develop into erythroid and T-lymphoid lineages, respectively. These findings support an instructive, but argue against a lineage restrictive role of C/EBPalpha in multipotent hematopoietic and thymic progenitors. We performed global gene expression profiling of double-sorted Cebpa/EYFP+ and Cebpa/EYFP- LSK cells of pooled Cebpa Cre/wt R26 EYFP reporter mice to identify differentially regulated genes in Cebpa+ versus Cebpa- LSK cells. RNA was isolated from three biological replicates of Cebpa/EYFP+ LSK cells and two biological replicates of Cebpa/EYFP- LSK cells. To determine if the identified genes were truly dependent on Cebpa expression, we also performed global gene expression profilling of Cebpa/EYFP+ and Cebpa/EYFP- fetal liver LSK cells of Cebpa Cre/fl R26 EYFP mice. Induction of Cebpa/Cre expression in these mice leads to Cre-mediated recombination of the floxed wt Cebpa allele resulting in a complete Cebpa knock-out. In this case, RNA was isolated from two biological replicates of either Cebpa/EYFP+ and Cebpa/EYFP- LSK cells. In addition, we included one biological replicate of Cebpa/EYFP+ and Cebpa/EYFP- fetal liver LSK cells of Cebpa Cre/wt R26 EYFP mice to determine the correlation of differentially regulated genes in bone marrow and fetal liver LSK cells.

Dataset Information

Transcription profiling of Mac-1+, c-Kit+ myeloid progenitor cells from mice homozygous for the p42-specific knock-out Cebpa mutation and independently transplanted with leukemic bone marrow, and from control mice heterozygous for Cebpa mutation

Publications

Modeling of C/EBPalpha mutant acute myeloid leukemia reveals a common expression signature of committed myeloid leukemia-initiating cells.

Similar Datasets

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