Extensive enhancer crosstalk controls PPARG2 activation during adipogenesis

Transcriptional master regulators drive cell fate transitions. Peroxisome proliferator-activated receptor γ (PPARγ) is the master regulator of adipogenesis, and its expression must therefore be tightly regulated and efficiently induced in response to adipogenic cues. Here we decipher the regulatory mechanisms of the highly connected enhancer community driving activation of the PPARG locus during adipocyte differentiation of human mesenchymal stem cells. By systematically deleting nine individual enhancers, spanning upstream, promoter-proximal, and downstream super-enhancer constituents, we demonstrate elaborate enhancer crosstalk in cis involving stabilization of C/EBPβ 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 expression. Non-coding genetic variants associated with cardiometabolic traits and predicted to regulate PPARG expression map to E + 102 and other essential enhancers in the community, thereby supporting the importance of these enhancers in human physiology and disease.