美国斯坦福大学Joseph C. Wu研究小组发现，人类iPSC-心肌细胞中的CRISPRi/a筛选发现，糖酵解激活是多柔比星引起的心脏毒性可药物化的靶点。2024年11月7日，国际知名学术期刊《细胞—干细胞》在线发表了这一成果。

研究人员展示了一种综合药物发现管线，结合了人类诱导多能干细胞（iPSC）来源的心肌细胞（iCM）、CRISPR干扰与激活（CRISPRi/a）双向汇集筛选以及小分子筛选。该研究旨在识别能减轻多柔比星引起的心脏毒性（DIC）而不影响其抗肿瘤效果的治疗靶点。

筛选结果揭示了多个先前未报道的候选基因，这些基因与DIC相关，其中包括碳酸酐酶12（CA12）。通过基因抑制CA12，可以通过改善细胞存活、肌原纤维结构完整性、收缩功能和钙离子处理，保护iCMs免受DIC的影响。

CA12拮抗剂Indisulam可以有效减轻iCM、工程化心脏组织和动物模型中的DIC。机制研究表明，多柔比星诱导的CA12通过促进心肌细胞中的糖酵解激活，干扰细胞代谢和功能，从而加剧了DIC。

综上所述，该研究为未来的药物发现提供了一条路线图，有望开发出更具靶向性、且副作用最小的治疗方案。

据了解，多柔比星由于其致命的心血管副作用而限制了其治疗效用。

附：英文原文

Title: CRISPRi/a screens in human iPSC-cardiomyocytes identify glycolytic activation as a druggable target for doxorubicin-induced cardiotoxicity

Author: Chun Liu, Mengcheng Shen, Yanxia Liu, Amit Manhas, Shane Rui Zhao, Mao Zhang, Nadjet Belbachir, Lu Ren, Joe Z. Zhang, Arianne Caudal, Masataka Nishiga, Dilip Thomas, Angela Zhang, Huaxiao Yang, Yang Zhou, Mohamed Ameen, Nazish Sayed, June-Wha Rhee, Lei S. Qi, Joseph C. Wu

Issue&Volume: 2024-11-07

Abstract: 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.

DOI: 10.1016/j.stem.2024.10.007

Source: https://www.cell.com/cell-stem-cell/abstract/S1934-5909(24)00369-2