Liu Chun, Shen Mengcheng, Liu Yanxia, Manhas Amit, Zhao Shane Rui, Zhang Mao, Belbachir Nadjet, Ren Lu, Zhang Joe Z, Caudal Arianne, Nishiga Masataka, Thomas Dilip, Zhang Angela, Yang Huaxiao, Zhou Yang, Ameen Mohamed, Sayed Nazish, Rhee June-Wha, Qi Lei S, Wu Joseph C
Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA; Department of Physiology and Cancer Center, Milwaukee, WI, USA; Department of Medicine and Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, USA.
Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA; Institute for Stem Cell Biology and Regenerative Medicine, Stanford, CA, USA; Department of Medicine (Division of Cardiology), Stanford, CA, USA.
Cell Stem Cell. 2024 Dec 5;31(12):1760-1776.e9. doi: 10.1016/j.stem.2024.10.007. Epub 2024 Nov 7.
Doxorubicin is limited in its therapeutic utility due to its life-threatening cardiovascular side effects. Here, we present an integrated drug discovery pipeline combining human induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iCMs), CRISPR interference and activation (CRISPRi/a) bidirectional pooled screens, and a small-molecule screening to identify therapeutic targets mitigating doxorubicin-induced cardiotoxicity (DIC) without compromising its oncological effects. The screens revealed several previously unreported candidate genes contributing to DIC, including carbonic anhydrase 12 (CA12). Genetic inhibition of CA12 protected iCMs against DIC by improving cell survival, sarcomere structural integrity, contractile function, and calcium handling. Indisulam, a CA12 antagonist, can effectively attenuate DIC in iCMs, engineered heart tissue, and animal models. Mechanistically, doxorubicin-induced CA12 potentiated a glycolytic activation in cardiomyocytes, contributing to DIC by interfering with cellular metabolism and functions. Collectively, our study provides a roadmap for future drug discovery efforts, potentially leading to more targeted therapies with minimal off-target toxicity.
由于其危及生命的心血管副作用，阿霉素的治疗效用受到限制。在此，我们展示了一个整合的药物发现流程，该流程结合了人类诱导多能干细胞（iPSC）衍生的心肌细胞（iCMs）、CRISPR干扰和激活（CRISPRi/a）双向混合筛选以及小分子筛选，以识别减轻阿霉素诱导的心脏毒性（DIC）而不损害其肿瘤学效应的治疗靶点。筛选揭示了几个先前未报道的与DIC相关的候选基因，包括碳酸酐酶12（CA12）。对CA12的基因抑制通过改善细胞存活、肌节结构完整性、收缩功能和钙处理来保护iCMs免受DIC影响。CA12拮抗剂茚地那韦可有效减轻iCMs、工程心脏组织和动物模型中的DIC。从机制上讲，阿霉素诱导的CA12增强了心肌细胞中的糖酵解激活，通过干扰细胞代谢和功能导致DIC。总的来说，我们的研究为未来的药物发现工作提供了路线图，可能会带来具有最小脱靶毒性的更有针对性的疗法。
