Extensive enhancer crosstalk governs the induction of PPARG2 expression during adipogenic lineage commitment [Capture-C]

Transcriptional master regulators drive cell fate transitions. Peroxisome proliferator- activated receptor ? (PPAR?) is the master regulator of adipogenesis, and its expression must therefore be tightly regulated and efficiently induced in response to adipogenic cues. Here we show that the PPARG locus is primed for activation in the mesenchymal stem cell state by a highly connected enhancer (HICE) community. By systematically deleting 9 individual enhancers, spanning upstream, promoter-proximal, and downstream super-enhancer constituents, we demonstrate elaborate enhancer crosstalk in cis involving stabilization of C/EBPß recruitment prior to chromatin remodeling. We show that the super-enhancer constituent E+102 plays a dual role in cis crosstalk and feedback activation and is obligate for activation of PPARG expresison. Non-coding genetic variants associated with cardiometabolic traits and predicted to regulate PPARG expression map E+102 and other essential enhancers in the community, thereby supporting the importance of these enhancers in human physiology and disease. Overall design: Chromatin interactions using Capture-C for PPARG2 TSS and selected enhancer in the PPARG locus during adipocyte differentiation of control and enhancer deletion human mesenchymal stem cells with iducible expression of Cas9 (hMSC-TERT4-iCas9-4).