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Deciphering regulation of FOXP3 expression in human conventional T cells

Umhoefer, J. M.; Arce, M. M.; Dajani, R.; Belk, J. A.; Mowery, C. T.; Nguyen, V.; Gowen, B. G.; Simeonov, D. R.; Curie, G. L.; Corn, J. E.; Chang, H. Y.; Marson, A.

2024-09-02 immunology
10.1101/2024.08.30.610436 bioRxiv
Show abstract

FOXP3 is a lineage-defining transcription factor (TF) for immune-suppressive regulatory T cells (Tregs). While mice exclusively express FOXP3 in Tregs, humans also transiently express FOXP3 in stimulated conventional CD4+ T cells (Tconvs). Mechanisms governing these distinct expression patterns remain unknown. Here, we performed CRISPR screens tiling the FOXP3 locus and targeting TFs in human Tregs and Tconvs to discover cis-regulatory elements (CREs) and trans-regulators of FOXP3. Tconv FOXP3 expression depended on a subset of Treg CREs and Tconv-selective positive (TcNS+) and negative (TcNS-) CREs. The CREs are occupied and regulated by TFs we identified as critical regulators of FOXP3. Finally, mutagenesis of murine TcNS- revealed that it is critical for restriction of FOXP3 expression to Tregs. We discover CRE and TF circuitry controlling FOXP3 expression and reveal evolution of mechanisms regulating a gene indispensable to immune homeostasis. HighlightsO_LIComprehensive CRISPR maps of CREs and TFs controlling FOXP3 in human Tregs and Tconvs C_LIO_LIKey TFs that control FOXP3 directly occupy and regulate CREs forming TF-CRE circuits C_LIO_LIA previously unknown negative CRE stringently restricts FOXP3 to Tregs in mice C_LI

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