CRISPR-Cas9 screening identified USP10 as a key regulatory gene and its mechanism for ovarian cancer to be sensitive to the PARP inhibitor olaparib
CRISPR-Cas9筛选鉴定USP10为卵巢癌对PARP抑制剂奥拉帕利敏感的关键调控基因及其机制

摘要

卵巢癌作为妇科恶性肿瘤中致死率最高的类型，其治疗面临耐药性频发的严峻挑战。PARP抑制剂奥拉帕利通过靶向DNA损伤修复缺陷（如BRCA突变）发挥抗肿瘤作用，但部分患者初治即不敏感或治疗后快速耐药，机制尚未完全明确。本研究采用CRISPR-Cas9全基因组敲除筛选技术，在卵巢癌细胞系中筛选调控奥拉帕利敏感性的关键基因，鉴定出泛素特异性蛋白酶10（USP10）为核心调控因子。功能验证显示，敲除USP10可显著增强卵巢癌细胞对奥拉帕利的敏感性，而USP10过表达则导致耐药；机制探究表明，USP10通过去泛素化稳定DNA修复蛋白RAD51，促进同源重组修复（HRR）通路激活，从而拮抗奥拉帕利诱导的DNA损伤积累。临床样本分析显示，卵巢癌组织中USP10高表达与患者奥拉帕利治疗响应差、无进展生存期短显著相关。本研究首次揭示USP10在卵巢癌奥拉帕利耐药中的关键作用，为优化PARP抑制剂治疗策略、开发联合治疗靶点提供了新的理论依据和实验基础。

Abstract

Ovarian cancer, the deadliest gynecological malignancy, faces significant challenges in treatment due to frequent drug resistance. PARP inhibitors like olaparib target DNA repair defects (e.g., BRCA mutations) to combat tumors, yet some patients show initial treatment resistance or rapid drug tolerance, with mechanisms remaining unclear. This study employed CRISPR-Cas9 genome-wide knockout screening to identify key genes regulating olaparib sensitivity in ovarian cancer cell lines, identifying ubiquitin-specific protease 10 (USP10) as a core regulatory factor. Functional validation demonstrated that USP10 knockout significantly enhances ovarian cancer cell sensitivity to olaparib, while overexpression induces drug resistance. Mechanistic analysis revealed that USP10 stabilizes DNA repair protein RAD51 through deubiquitination, activating the homologous recombination repair (HRR) pathway and counteracting olaparib-induced DNA damage accumulation. Clinical data showed strong correlation between high USP10 expression in ovarian cancer tissues and poor olaparib response and short progression-free survival. This study provides the first revelation of USP10's critical role in olaparib resistance, offering new theoretical foundations and experimental basis for optimizing PARP inhibitor strategies and developing combination therapy targets.