Suga Takaomi, Kitani Tomoya, Kogure Masaya, Oishi Masatsugu, Ito Fumiaki, Hoshino Atsushi, Ogata Takehiro, Ikeda Koji, Matoba Satoaki
Department of Cardiovascular Medicine, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan.
Department of Pathology and Cell Regulation, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
Cardiovasc Res. 2025 May 6;121(4):601-613. doi: 10.1093/cvr/cvaf022.
Doxorubicin (DOX) is one of the most effective chemotherapeutic agents for various types of cancers. However, DOX often causes cardiotoxicity, which is referred to as DOX-induced cardiomyopathy (DIC). Despite extensive research, only a limited number of effective treatments are currently available. In this study, we aimed to identify a potential therapeutic target for DIC by preventing DOX-induced cell injury in cardiomyocytes.
We performed a kinome-wide CRISPR gene knockout screen in human cardiomyocytes derived from pluripotent stem cells (hPSC-CMs) and identified a member of the STE20 kinase family, thousand and one amino acid protein kinase 1 (TAOK1) as a potential regulator of DOX-induced cardiomyocyte death. Using CRISPR-mediated gene knockout and small interfering RNA-mediated gene knockdown, we demonstrated that TAOK1 suppression improved DOX-induced cardiomyocyte death and dysfunction, including sarcomere disarray, contractile dysfunction, DNA damage, and mitochondrial dysfunction in hPSC-CMs. Transcriptome analysis using RNA-seq also showed that DOX-induced mitochondrial dysfunction was attenuated by TAOK1 suppression. In contrast to the protective role of TAOK1 against DOX toxicity in cardiomyocytes, TAOK1 suppression did not induce DOX resistance in human cancer cell lines. DOX-induced activation of p38 mitogen-activated protein kinase (MAPK) was markedly attenuated in TAOK1-knockout hPSC-CMs. Furthermore, DOX-induced cardiomyocyte death and disruption of mitochondrial membrane potential were augmented by TAOK1 overexpression, which was partially attenuated by an inhibitor or knockdown of p38 MAPK or an apoptosis inhibitor. Finally, we demonstrated that TAOK1 suppression using adeno-associated virus (AAV)-mediated gene silencing attenuated DOX-induced myocardial damage, including myocardial fibrosis, apoptosis, and cardiomyocyte atrophy, resulting in improved cardiac function in a mouse model of DIC.
Our results indicate that TAOK1 suppression is a promising therapeutic approach for treating DIC in patients with cancer and highlight the advantages of hPSC-CMs as a platform to study drug-induced cardiotoxicity.
阿霉素（DOX）是治疗各类癌症最有效的化疗药物之一。然而，DOX常导致心脏毒性，即所谓的DOX诱导的心肌病（DIC）。尽管进行了广泛研究，但目前有效的治疗方法仍然有限。在本研究中，我们旨在通过预防DOX诱导的心肌细胞损伤来确定DIC的潜在治疗靶点。
我们在源自多能干细胞的人心肌细胞（hPSC-CMs）中进行了全激酶组CRISPR基因敲除筛选，并确定STE20激酶家族成员之一的千与一氨基酸蛋白激酶1（TAOK1）是DOX诱导的心肌细胞死亡的潜在调节因子。使用CRISPR介导的基因敲除和小干扰RNA介导的基因敲低，我们证明抑制TAOK1可改善DOX诱导的心肌细胞死亡和功能障碍，包括hPSC-CMs中的肌节紊乱、收缩功能障碍、DNA损伤和线粒体功能障碍。使用RNA测序进行的转录组分析还表明，抑制TAOK1可减轻DOX诱导的线粒体功能障碍。与TAOK1对心肌细胞DOX毒性的保护作用相反，抑制TAOK1不会在人癌细胞系中诱导DOX耐药。在TAOK1敲除的hPSC-CMs中，DOX诱导的p38丝裂原活化蛋白激酶（MAPK）激活明显减弱。此外，TAOK1过表达增强了DOX诱导的心肌细胞死亡和线粒体膜电位的破坏，p38 MAPK抑制剂或敲低或凋亡抑制剂可部分减弱这种增强作用。最后，我们证明使用腺相关病毒（AAV）介导的基因沉默抑制TAOK1可减轻DOX诱导的心肌损伤，包括心肌纤维化、细胞凋亡和心肌细胞萎缩，从而改善DIC小鼠模型的心脏功能。
我们的结果表明，抑制TAOK1是治疗癌症患者DIC的一种有前景的治疗方法，并突出了hPSC-CMs作为研究药物诱导心脏毒性平台的优势。
