Cars2‐Mediated Cysteine Catabolism Drives Brown Fat Development and Thermogenesis Through Persulfidating EBF2

ABSTRACT Targeting transcriptional regulatory complexes represents a promising strategy for activating thermogenic fat and treating obesity. In this study, we focused on identifying critical, specific metabolic pathways enriched during brown adipocyte differentiation. Through analysis of metabolomics and RNA sequencing data from precursor and mature brown and white adipocytes, we revealed that the cysteine catabolism pathway constitutes a unique metabolic process that is significantly upregulated during brown adipocyte differentiation. We further demonstrated that the cysteine persulfidation synthase Cars2, which 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, resulting in mice exhibiting reduced thermogenesis and energy expenditure. More importantly, Cars2 generates cysteine persulfide and its derivative H ₂ S cell‐autonomously induces brown adipogenesis and enhances uncoupled respiration through persulfidation of EBF2. Mechanistically, persulfidated EBF2 facilitates its interaction with PPARγ or BRG1, enhancing recruitment of the EBF2‐PPARγ complex to browning gene promoters, thereby driving brown fat development and boosting thermogenic activity. Furthermore, treatment with the Cars2 coenzyme PLP or an H ₂ S donor enhances brown adipocyte function and alleviates obesity progression in mice fed a high‐fat diet, indicating a novel metabolite‐based strategy for obesity treatment.