Disulfidptosis-Induced Chondrocyte-Macrophage Crosstalk via GYS1/CCND1/NOD2 Axis Promotes Osteoarthritis Progression

Qing Sun,1,* Zhihui Wei,1,* Gaohai Shao,1 Mengfan Yang,1 Xianyan Zeng,2 Xiang Hong,1 Rong Gou,3 Han Zhang,4 Yining Zhang,5 Shuheng Dong,1 Yiming Qu,1 Yujiao Liu1,5 1Department of Orthopedics, Affiliated Yongchuan Hospital of Chongqing Medical University, Chongqing, 402160, People’s Republic of China; 2College of Pharmacy, Chongqing Medical University, Chongqing, 402160, People’s Republic of China; 3Affiliated Cancer Hospital of Chongqing University, Chongqing, 400000, People’s Republic of China; 4College of Biomedical Engineering, Hubei University of Medicine, Hubei, 442000, People’s Republic of China; 5Laboratory of Skeletal Development and Regeneration, Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yujiao Liu, Department of Orthopedics, Affiliated Yongchuan Hospital of Chongqing Medical University, Chongqing, 402160, People’s Republic of China, Email [email protected] Yiming Qu, Department of Orthopedics, Affiliated Yongchuan Hospital of Chongqing Medical University, Chongqing, 402160, People’s Republic of China, Email [email protected]: To investigate the potential role and underlying mechanisms of disulfidptosis, a novel form of regulated cell death, in the pathogenesis of degenerative osteoarthritis (OA), and to evaluate its association with M1-type macrophage infiltration and diagnostic biomarker potential.Methods: Human articular cartilage samples from OA patients and non-OA controls were analyzed using high-throughput RNA sequencing to identify disulfidptosis-related gene expression patterns. Immunohistochemical assays were performed to validate key molecular signatures in chondrocytes. Differentially expressed genes (DEGs) were screened to identify candidates linking disulfidptosis to M1 macrophage biology. Functional correlation analyses were conducted to explore gene–immune cell interactions. Glucose starvation was applied to induce disulfidptosis in ATDC5 chondrocyte line; a co-culture system with RAW264.7 macrophage cell line was established to validate their functional roles and underlying mechanisms.Results: OA chondrocytes exhibited a low-glucose metabolic state and elevated SLC7A11 (a cystine/glutamate transporter implicated in disulfidptosis) expression, consistent with a disulfidptosis signature, and showed increased immune infiltration of M1-type macrophages. Among the DEGs, Glycogen synthase 1 (GYS1) emerged as a key disulfidptosis-related gene directly regulating transcriptional programs in M1 macrophages. Functional analyses suggested that disulfidptosis in chondrocytes may indirectly promote M1 macrophage-mediated immune infiltration through cyclin D1 (CCND1) and nucleotide-binding oligomerization domain-containing protein 2 (NOD2), thereby contributing to OA progression.Conclusion: Disulfidptosis in chondrocytes is associated with M1 macrophage immune infiltration and could drive OA progression. Mechanism genes like GYS1/CCND1/NOD2 have diagnostic potential and could be novel biomarkers and therapeutic targets for OA. These findings highlight the translational potential of targeting disulfidptosis-related pathways to improve OA management and patient outcomes. Keywords: disulfidptosis, macrophages, osteoarthritis, GYS1, chondrocyte