CarS2-mediated cysteine catabolism drives brown fat development and thermogenesis through persulfidating EBF2

Targeting transcriptional regulatory complexes is a promising strategy to activate thermogenic fat and treat obesity. In this study, we focused on identifying critical and specific metabolic pathways enriched during brown adipocyte differentiation. By analyzing metabolomics and RNA sequencing results from precursor and mature brown and white adipocytes, we revealed cysteine catabolism pathway is a unique metabolic process that is highly increased in brown adipocyte differentiation. Then, we demonstrated cysteine persulfidation synthase CarS2 is directly induced by EBF2, mediates cysteine catabolism and triggers brown fat protein persulfidation both in vitro and in vivo. Loss of CarS2 in thermogenic fat blocks brown fat formation in both male and female mice, and CarS2 depletion in mice reduces thermogenesis and energy expenditure. More importantly, CarS2 generates cysteine persulfide, and its derivative H2S cell-autonomously induces brown adipogenesis and enhances uncoupled respiration by persulfidating EBF2. Mechanistically, persulfidated EBF2 facilitates its interaction with PPARγ or BRG1, enhancing the recruitment of the EBF2-PPARγ complex to the browning gene promoter, thus driving brown fat development and boosting thermogenic activity. Furthermore, treatment with CarS2 coenzyme PLP or H2S donor elevates brown adipocyte function and ameliorates obesity progression in mice under HFD feeding, indicating a novel metabolite-based strategy for obesity treatment.