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RNA-seq profiling of mammary epithelial cell populations from Brca2 knockout and wildtype mice

ABSTRACT: RNA-seq profiling of mammary epithelial cell populations from Brca2 knockout and wildtype mice

PROVIDER: PRJNA973059 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24583306_1.fastq.gz	Fastqsanger.gz
	SRR24583306_2.fastq.gz	Fastqsanger.gz
	SRR24583307_1.fastq.gz	Fastqsanger.gz
	SRR24583307_2.fastq.gz	Fastqsanger.gz
	SRR24583308_1.fastq.gz	Fastqsanger.gz

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Project description:CCAAT/enhancer binding protein beta (C/EBPb) is a member of a family of highly conserved transcription factors that regulates numerous genes involved in proliferation and differentiation in a variety of tissues. C/EBPb is deregulated in human breast cancer and germline deletion of this gene results in multiple defects in mammary gland development. We hypothesized that C/EBPb regulates mammary stem cell self-renewal, maintenance and/or differentiation through the regulation of multiple target genes that coordinate mammary gland development. Utilizing both a germline knockout mouse model and a conditional knockout strategy, we demonstrated that mammosphere formation was significantly decreased in C/EBPb-deficient mammary epithelial cells (MECs). The ability of C/EBPb-deleted MECs to regenerate the mammary gland in vivo was severely impaired when transplanted at limiting dilution. Furthermore, serial transplantation of C/EBPb-null mammary tissue resulted in decreased outgrowth potential when compared to wildtype, and an early senescence phenotype. Flow cytometric analysis revealed that C/EBPb-null MECs contain a lower frequency of repopulating stem cells accompanied by an increase in committed, differentiated luminal cells as compared to wildtype. Microarray analysis of stem/progenitor cell populations was performed and revealed an alteration in cell fate specification in C/EBPb-null glands, exemplified by the aberrant expression of basal markers in the luminal cell compartment. Collectively, our studies demonstrate that C/EBPb is a critical regulator of mammary stem cell differentiation, and an important determinant of luminal cell fate specification. Experiment Overall Design: To identify potential signaling pathways regulated by C/EBPb in stem/progenitor cells, microarray analysis was performed on two stem/progenitor cell subpopulations. For this analysis, subpopulations defined by LIN-CD24+CD29hi and LIN-CD24hiCD29lo were FACS sorted from wildtype and germline C/EBPb-/- glands, and RNA was isolated from each group.

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RNA-seq profiling of mammary epithelial cell populations from Brca2 knockout and wildtype mice

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