Abstract

Research Question Does pyruvate carboxylase (PC) drive endometriosis progression and, if so, what is the underlying mechanism? Design We examined PC expression in clinical endometrial specimens via immunohistochemistry. After PC knockdown or overexpression in immortalized human endometriotic stromal cells (ihESCs), cell proliferation and migration were assessed using 5-ethynyl-2′-deoxyuridine (EdU), colony formation and Transwell assays. Quasi-targeted metabolomics, RNA sequencing and western blotting were adopted to explore downstream molecular changes, and an allogeneic mouse endometriosis model was applied for in vivo functional verification. Results First, we found that PC expression was significantly upregulated in eutopic and ectopic endometrial tissues of patients with endometriosis compared with controls (p=0.022 and p<0.001, respectively). Then, in vitro and in vivo experiments showed that PC promoted the progression of endometriosis. In addition, quasi-targeted metabolomics showed that PC knockdown decreased the abundance of glycolytic metabolites in ihESCs. Furthermore, RNA sequencing and western blotting identified that PC enhanced the activity of the phosphoinositide 3-kinase (PI3K)-AKT pathway. Using AKT agonist (SC79) and inhibitor (AZD5363), we found that PC modulates glycolytic enzyme expression and endometriosis progression through the AKT pathway. Finally, PC inhibitors ZY-444 and octyl gallate attenuated the progression of endometriosis. Conclusions PC is significantly upregulated in eutopic and ectopic endometrial tissues from endometriosis patients, and modulates glycolytic enzyme expression as well as endometriosis progression via activation of the AKT pathway. Our findings provide mechanistic insight into the metabolic reprogramming underlying endometriosis pathogenesis.