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Circular RNA (circRNA) is expected to provide a long-lasting therapy for OA. However, the involvement of the circRNA-associated competitive endogenous RNA network in chondrocyte senescence induced by mechanical overloading remains unestablished. A mechanical overloading-induced chondrocyte senescence model in human primary chondrocytes is constructed, and differences in the expression of circRNAs and miRNAs were analyzed. The biological roles of circKIAA0586\/miR-335-5p in chondrocyte senescence and OA progression under mechanical overloading and its downstream targets were determined using gain- and loss-of-function experiments in various biochemical assays in human chondrocytes. The in\u00a0vivo effects of circKIAA0586 overexpression were also determined in destabilization of the medial meniscus (DMM) OA mice and aged spontaneous OA mice. The mechanical overloading-induced chondrocyte senescence was aggravated by miR-335-5p or circKIAA0586 knockdown. Accumulated DNA damage response was observed following mechanical overloading, which reduced after miR-335-5p inhibition or circKIAA0586 supplementation. MiR-335-5p was regulated by circKIA0586 adsorption. HELLS was prominently down-regulated following mechanical overloading treatment. Moreover, miR-335-5p bound to lymphoid-specific helicase (HELLS) mRNA during mechanical overloading was demonstrated to mediate the nonhomologous end joining (NHEJ) pathway, thereby inducing DNA damage and senescence. In addition, the senescence delaying and cartilage protective functions of circKIAA0586 and HELLS were validated in DMM OA mice and aged spontaneous OA mice. Our findings suggest that miR-335-5p, which escapes circKIAA0586 adsorption, facilitates mechanical overloading-induced chondrocyte senescence and OA progression by impairing the NHEJ pathway through HELLS inhibition. Overall, targeting circKIAA0586\/miR-335-5p\/HELLS signaling provides a novel therapeutic approach for OA.<\/jats:p>","DOI":"10.34133\/research.0694","type":"journal-article","created":{"date-parts":[[2025,4,16]],"date-time":"2025-04-16T13:01:01Z","timestamp":1744808461000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark_01","source":"Crossref","is-referenced-by-count":10,"title":["MiR-335-5p Escaped from CircKIAA0586 Adsorption Contributes to Mechanical Overloading-Induced Cartilage Degeneration by Targeting Lymphoid-Specific Helicase"],"prefix":"10.34133","volume":"8","author":[{"given":"Haoyu","family":"Xie","sequence":"first","affiliation":[{"name":"Department of Joint Surgery, Center for Orthopaedic Surgery, \rThe Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China."},{"name":"Department of Orthopedics, Orthopedic Hospital of Guangdong Province, Academy of Orthopedics\u00b7Guangdong Province, \rThe Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China."},{"name":"The Third School of Clinical Medicine, \rSouthern Medical University, Guangzhou 510630, China."},{"name":"Guangdong Provincial Key Laboratory of Bone and Joint Degeneration Diseases","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuheng","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Rehabilitation Medicine, Xijing Hospital, \rFourth Military Medical University, Xi \u2019an 710032, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianying","family":"Pan","sequence":"additional","affiliation":[{"name":"Department of Joint Surgery, Center for Orthopaedic Surgery, \rThe Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China."},{"name":"Department of Orthopedics, Orthopedic Hospital of Guangdong Province, Academy of Orthopedics\u00b7Guangdong Province, \rThe Third Affiliated Hospital of Southern Medical University, 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