Integrated single-cell and experimental validation links effector chondrocyte-associated oxidative stress to EIF6-related p47phox–NOX2 signaling in osteoarthritis

作者：Shuai Yuan, Huirun Chen, Jiaxue Wei, Jinxian Tan, Shuai Liu, Zhenyang Fu · 发表于：Frontiers in Physiology · 年份：2026 · DOI：10.3389/fphys.2026.1882484 · 研究领域：Osteoarthritis Treatment and Mechanisms、Single-cell and spatial transcriptomics、Bioinformatics and Genomic Networks

Background Oxidative stress is closely involved in osteoarthritis (OA), but the cell-type-specific oxidative stress landscape and related regulatory molecules in human cartilage remain incompletely defined. This study aimed to identify effector chondrocyte (EC)-associated oxidative stress features in OA and to prioritize candidate molecules linked to p47phox–NOX2-related ROS responses. Methods Public human cartilage single-cell RNA sequencing datasets were integrated to construct an OA cartilage atlas. Oxidative stress activity was evaluated using multiple scoring methods, and high oxidative stress (HOS) cells were defined according to the top 10% of integrated oxidative stress scores. Cell-state and communication features were examined using CytoTRACE and CellChat. High-dimensional weighted gene co-expression network analysis, machine learning, protein–protein interaction network analysis, summary-data-based Mendelian randomization, and molecular docking were combined to prioritize candidate genes. EIF6 and YWHAB were examined in an inflammatory EC-like model, and EIF6-associated p47phox/TGF-β responses were further assessed in DMM-induced OA mouse tissues. Results Single-cell analysis identified an EC population enriched in OA cartilage. ECs showed relatively high oxidative stress activity and represented the largest component of HOS cells. The proportion of ECs among HOS cells increased in OA samples, and HOS cells were concentrated near the EC-enriched region in the UMAP ...