Expression Analysis of Mitochondrial Energy Metabolism−Related Genes Identifies IRS2 as a Key Modulator in M2 Synovial Macrophages of Osteoarthritis
Yunlong Yang, Nianlong Zhang, Xuyang Li, Enbei Xie, Yangyu Wu, Jianlin ZhouBackground: Mitochondrial bioenergetic dysregulation disrupts immune−metabolic homeostasis and promotes pro−inflammatory microenvironments in osteoarthritis (OA) synovitis. However, the mechanistic contributions of mitochondrial energy metabolism to synovitis pathogenesis in OA remain poorly defined. Methods: We analyzed mitochondrial energy metabolism−related genes (MEMRGs) in OA synovitis by integrating transcriptomic data from OA synovial tissues (GSE55235, GSE55457). LASSO regression and maximal clique centrality (MCC) algorithms were applied to identify hub genes, and single−cell RNA sequencing (GSE152805) was used to examine cell−type−specific expression patterns. Functional validation was performed in IRS2−knockdown THP−1 macrophages. Results: We identified 22 mitochondrial energy metabolism−related differentially expressed genes (MEMR−DEGs), which were enriched in the AMPK signaling, glucagon signaling, and insulin signaling pathways. Four hub genes (FOXO3, FASN, PTGS2, IRS2) were identified, and their expression was negatively correlated with synovial macrophage infiltration. Single−cell RNA sequencing revealed that IRS2 was specifically upregulated in a synovial macrophage cluster. Functional studies in IRS2−knockdown THP−1 macrophages demonstrated that IRS2 deficiency impaired IL−4−induced M2 macrophage polarization and reduced mitochondrial membrane potential and ATP synthesis, which was mediated by the suppression of the AKT/FOXO1 signaling. Conclusions: IRS2 potentially influences mitochondrial energy metabolism, as evidenced by the maintenance of mitochondrial membrane potential and ATP synthesis, via the AKT/FOXO1 signaling pathways to maintain synovial macrophage M2 polarization homeostasis. These findings provide novel molecular targets for addressing immune−metabolic pathways in OA therapy.