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<jats:p>Cisplatin\u2010induced ototoxicity remains a clinical dilemma with limited mechanistic understanding and no food and drug administration (FDA)\u2010approved therapies. Despite emerging roles of small extracellular vesicles (sEV) in drug ototoxicity, their molecular cargo profiles and causal roles to cisplatin\u2010induced ototoxicity are unexplored. This study systematically investigates sEV derived from cochlear explants treated with cisplatin (Cis\u2010sEV) and controls (Ctrl\u2010sEV) using multi\u2010omics profiling. Through small RNA sequencing, 83 differentially expressed microRNAs (miRNAs) are identified in Cis\u2010sEV compared to Ctrl\u2010sEV. Notably, mmu\u2010miR\u201034a\u20105p, mmu\u2010miR\u2010140\u20105p, mmu\u2010miR\u201015b\u20105p, mmu\u2010miR\u201025\u20103p, and mmu\u2010miR\u2010339\u20105p are significantly upregulation in Cis\u2010sEVs. Predicted target pathways of these differentially expressed miRNAs are enriched in apoptosis, inflammation, and cellular damage, indicating their potential involvement in cisplatin\u2010induced cochlear damage. LC\u2010MS\/MS analysis reveals 90 upregulated and 150 downregulated proteins in Cis\u2010sEV, with many involved in damage response. Specifically, CLTC, CCT2, ANXA6, and HSPA8 are uniquely upregulated proteins in Cis\u2010sEV, and CLTC and ANXA6 are exclusively co\u2010localized in hair cells (HCs) post\u2010cisplatin exposure, suggesting that Cis\u2010sEV originate primarily from damaged HCs. Moreover, CLTC in sEV may serve as a potential biomarker for cisplatin\u2010induced ototoxicity as verified in both in vitro and in vivo models. This study provides novel insights into the molecular mechanisms of cisplatin\u2010induced ototoxicity and identifies potential biomarker and therapeutic targets.<\/jats:p>","DOI":"10.1002\/advs.202502627","type":"journal-article","created":{"date-parts":[[2025,5,24]],"date-time":"2025-05-24T07:05:57Z","timestamp":1748070357000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Small Extracellular Vesicles Orchestrate Cisplatin\u2010Induced Ototoxicity: Potential Biomarker and Targets Discovery"],"prefix":"10.1002","volume":"12","author":[{"given":"Jingru","family":"Ai","sequence":"first","affiliation":[{"name":"State Key Laboratory of Digital Medical Engineering Department of Otolaryngology Head and Neck Surgery Zhongda Hospital School of Life Sciences and Technology Advanced Institute for Life and Health Jiangsu Province High\u2010Tech Key Laboratory for Bio\u2010Medical Research Southeast University  Nanjing 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