WWP2 underlies ROS-induced granulosa cell apoptosis by promoting ubiquitination of BAK in polycystic ovary syndrome.

Granulosa cell (GC) apoptosis is intrinsically linked to the ovarian dysfunction of polycystic ovary syndrome (PCOS). Although oxidative stress and apoptosis in GCs have been detected in PCOS patients, how reactive oxygen species (ROS) links to GC apoptosis in PCOS remains to be further elucidated. Here, by integrating public single-cell RNA-seq data with clinical GC sample validation, we found that the expression of the E3 ubiquitin ligase WWP2 was significantly reduced, whereas its role in PCOS has not been previously reported. Notably, we first demonstrated that WWP2 overexpression can effectively antagonize mitochondrial apoptosis and ROS in KGNs. Mechanistically, oxidative stress weakened the interaction between WWP2 and BAK and reduced WWP2 expression, thereby suppressing BAK ubiquitination at Lys113. This inhibition impaired proteasomal degradation and consequently increased BAK protein levels. Consistently, disrupting BAK ubiquitination (BAK-K113R mutant) or knocking down WWP2 facilitated KGN apoptosis, and genetic ablation of Wwp2 in PCOS mice further aggravated GC apoptosis and hormonal disturbances. This study elucidates the molecular mechanism by which oxidative stress modulates GC mitochondrial apoptosis through WWP2-mediated BAK ubiquitination, and establishes WWP2 as a potential therapeutic target for PCOS.