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.

中文翻译：

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人 iPSC 心肌细胞中的 CRISPRi/a 筛选将糖酵解激活确定为阿霉素诱导的心脏毒性的可成药靶标


由于阿霉素会危及生命的心血管副作用，其治疗效用受到限制。在这里，我们提出了一个综合的药物发现管道，结合了人诱导多能干细胞 （iPSC） 衍生的心肌细胞 （iCM）、CRISPR 干扰和激活 （CRISPRi/a） 双向混合筛选，以及小分子筛选，以确定减轻阿霉素诱导的心脏毒性 （DIC） 的治疗靶点，而不会影响其肿瘤作用。筛选揭示了几个以前未报道的导致 DIC 的候选基因，包括碳酸酐酶 12 （CA12）。CA12 的遗传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 iCM 免受 DIC 的侵害。Indisulam 是一种 CA12 拮抗剂，可有效减弱 iCM、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来促进 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，有可能带来更具靶向性的治疗，同时将脱靶毒性降至最低。