Du Jie, Chen Fuqiang, Du Chenghong, Zhao Wenna, Chen Zihan, Ding Zhenhua, Zhou Meijuan
Jiangmen Central Hospital, Affiliated Jiangmen Hospital of Sun Yat-sen University, Jiangmen, Guangdong, China.
Department of Radiation Medicine, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, Guangdong, China.
Cell Death Discov. 2024 Oct 11;10(1):434. doi: 10.1038/s41420-024-02201-1.
DNA damage is considered to be a potentially unifying driver of ageing, and the stalling of DNA damage repair accelerates the cellular senescence. However, augmenting DNA repair has remained a great challenge due to the intricate repair mechanisms specific for multiple types of lesions. Herein, we miniaturized our modified detecting system for homologous recombination (HR) into a 96-well-based platform and performed a high-throughput chemical screen for FDA-approved drugs. We uncovered that amodiaquine could significantly augment HR repair at the noncytotoxic concentration. Further experiments demonstrated that amodiaquine remarkably suppressed stress-induced premature cellular senescence (SIPS), as evidenced by senescence-associated beta-galactosidase (SA-β-gal) staining or senescence-related markers p21 and p16, and the expression of several cytokines. Mechanistic studies revealed that the stimulation of HR repair by amodiaquine might be mostly attributable to the promotion of SIRT1 at the transcriptional level. Additionally, SIRT1 depletion abolished the amodiaquine-mediated effects on DNA repair and cellular senescence, indicating that amodiaquine delayed the onset of SIPS via a SIRT1-dependent pathway. Taken together, this experimental approach paved the way for the identification of compounds that augment HR activity, which could help to underscore the therapeutic potential of targeting DNA repair for treating aging-related diseases.
DNA损伤被认为是衰老潜在的统一驱动因素，而DNA损伤修复的停滞会加速细胞衰老。然而，由于多种类型损伤所特有的复杂修复机制，增强DNA修复仍然是一个巨大的挑战。在此，我们将改良的同源重组（HR）检测系统小型化到基于96孔的平台上，并对FDA批准的药物进行了高通量化学筛选。我们发现阿莫地喹在无细胞毒性浓度下可显著增强HR修复。进一步的实验表明，阿莫地喹能显著抑制应激诱导的早衰（SIPS），衰老相关β-半乳糖苷酶（SA-β-gal）染色或衰老相关标志物p21和p16以及几种细胞因子的表达均证明了这一点。机制研究表明，阿莫地喹对HR修复的刺激作用可能主要归因于其在转录水平上对SIRT1的促进作用。此外，SIRT1的缺失消除了阿莫地喹对DNA修复和细胞衰老的介导作用，表明阿莫地喹通过SIRT1依赖性途径延迟了SIPS的发生。综上所述，这种实验方法为鉴定增强HR活性的化合物铺平了道路，这有助于强调靶向DNA修复治疗衰老相关疾病的治疗潜力。
