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Critical Role of Runx Transcription Factors in Maintaining Epididymal Epithelial Differentiation [RNA-seq II]

ABSTRACT: Critical Role of Runx Transcription Factors in Maintaining Epididymal Epithelial Differentiation [RNA-seq II]

PROVIDER: PRJNA989043 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25068714_1.fastq.gz	Fastqsanger.gz
	SRR25068714_2.fastq.gz	Fastqsanger.gz
	SRR25068715_1.fastq.gz	Fastqsanger.gz
	SRR25068715_2.fastq.gz	Fastqsanger.gz
	SRR25068716_1.fastq.gz	Fastqsanger.gz

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Project description:Runx transcription factors are essential for generating functional T cells starting from the earliest stages in thymic T-development. To evaluate how Runx factors instruct T cell development, we perturbed Runx activities using 1) CRISPR-Cas9, simultaneously deleting two predominant, redundant Runx factors, Runx1 and Runx3, and 2) Runx1-overepxression at a stage before pro-T cells commit to a T cell fate. Then single-cell transcriptome analysis was performed. Runx1/Runx3 knockout (KO) vs. Runx1 overexpression (OE) resulted in contrasting deviations from the normal developmental trajectory. The separation of both perturbation conditions from the controls implied that Runx factors regulate gene networks in individual cells to control separate pathways instead of changing distributions of cells among different stages along the control pathway. Notably, a common core set of genes responded to both KO and OE, in opposite directions. Runx activation target genes were selectively enriched for T-identity program and innate lymphoid cell program genes, whereas inhibition targets were significantly associated with stem/progenitor program and myeloid lineage pathways. Pseudotime inference analysis suggested that Runx perturbations also substantially regulate developmental speed, as Runx KO significantly slowed down developmental progression, whereas Runx1 OE resulted in pronounced acceleration. This interpretation was largely supported by Runx activities regulating key transcription factors involved in establishing distinct gene network modules during T cell development. Indeed, further analysis showed that Runx target genes get combinatorial inputs from these Runx-regulated transcription factors as well as from Runx factors themselves. Together, our data suggest that Runx transcription factors drive early T developmental progression by launching gene expression modules associated with T-identity and innate lymphoid cells through mediating other transcription factor cooperativities.

Dataset Information

Critical Role of Runx Transcription Factors in Maintaining Epididymal Epithelial Differentiation [RNA-seq II]

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