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A fully validated blood stem/progenitor cell regulatory network reveals mechanisms of cell state stabilisation

ABSTRACT: The aim of this study was to determine the genomic binding sites of important haematopoietic transcription factors in haematopoietic cell lines.

ORGANISM(S): Mus musculus

PROVIDER: GSE69776 | GEO | 2016/03/17

SECONDARY ACCESSION(S): PRJNA286742

REPOSITORIES: GEO

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A fully validated blood stem/progenitor cell regulatory network reveals mechanisms of cell state stabilisation

Project description:The aim of this study was to determine the genomic binding sites of important haematopoietic transcription factors in haematopoietic cell lines. 416Bs cells were fixed with either 1 or 0.4 % formaldehyde for 10 mins. Chromatin was isolated, sonicated for 7 mins (30 sec on, 30 sec off), and specific antibodies were used to pull down the transcription factors of interest after a pre-clearing step. Chromatin was washed, de-crosslinked, amplified, size selected by gel purification and sequenced.

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Project description:Th17 cells have critical roles in mucosal defense and are major contributors to inflammatory disease. Their differentiation requires the nuclear hormone receptor RORγt working with multiple other essential transcription factors (TFs). We have used an iterative systems approach, combining genome-wide TF occupancy, expression profiling of TF mutants, and expression time series to delineate the Th17 global transcriptional regulatory network. We find that cooperatively-bound BATF and IRF4 contribute to initial chromatin accessibility, and with STAT3 initiate a transcriptional program that is then globally tuned by the lineage-specifying TF RORγt, which plays a focal deterministic role at key loci. Integration of multiple datasets allowed inference of an accurate predictive model that we computationally and experimentally validated, identifying multiple new Th17 regulators, including Fosl2, a key determinant of cellular plasticity. This interconnected network can be used to investigate new therapeutic approaches to manipulate Th17 functions in the setting of inflammatory disease. 143 RNA-seq, 83 ChIP-seq, 65 ChIP-seq controls, and 16 FAIRE-seq

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