通过电起搏评估的患者特异性人诱导多能干细胞模型验证了S107作为儿茶酚胺能多形性室性心动过速潜在治疗药物的有效性。
Patient-Specific Human Induced Pluripotent Stem Cell Model Assessed with Electrical Pacing Validates S107 as a Potential Therapeutic Agent for Catecholaminergic Polymorphic Ventricular Tachycardia.
作者信息
Sasaki Kenichi, Makiyama Takeru, Yoshida Yoshinori, Wuriyanghai Yimin, Kamakura Tsukasa, Nishiuchi Suguru, Hayano Mamoru, Harita Takeshi, Yamamoto Yuta, Kohjitani Hirohiko, Hirose Sayako, Chen Jiarong, Kawamura Mihoko, Ohno Seiko, Itoh Hideki, Takeuchi Ayako, Matsuoka Satoshi, Miura Masaru, Sumitomo Naokata, Horie Minoru, Yamanaka Shinya, Kimura Takeshi
机构信息
Department of Cardiovascular Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan.
Kyoto University iPS Cell Research and Application, Kyoto, Japan.
出版信息
PLoS One. 2016 Oct 20;11(10):e0164795. doi: 10.1371/journal.pone.0164795. eCollection 2016.
INTRODUCTION
Human induced pluripotent stem cells (hiPSCs) offer a unique opportunity for disease modeling. However, it is not invariably successful to recapitulate the disease phenotype because of the immaturity of hiPSC-derived cardiomyocytes (hiPSC-CMs). The purpose of this study was to establish and analyze iPSC-based model of catecholaminergic polymorphic ventricular tachycardia (CPVT), which is characterized by adrenergically mediated lethal arrhythmias, more precisely using electrical pacing that could promote the development of new pharmacotherapies.
METHOD AND RESULTS
We generated hiPSCs from a 37-year-old CPVT patient and differentiated them into cardiomyocytes. Under spontaneous beating conditions, no significant difference was found in the timing irregularity of spontaneous Ca2+ transients between control- and CPVT-hiPSC-CMs. Using Ca2+ imaging at 1 Hz electrical field stimulation, isoproterenol induced an abnormal diastolic Ca2+ increase more frequently in CPVT- than in control-hiPSC-CMs (control 12% vs. CPVT 43%, p<0.05). Action potential recordings of spontaneous beating hiPSC-CMs revealed no significant difference in the frequency of delayed afterdepolarizations (DADs) between control and CPVT cells. After isoproterenol application with pacing at 1 Hz, 87.5% of CPVT-hiPSC-CMs developed DADs, compared to 30% of control-hiPSC-CMs (p<0.05). Pre-incubation with 10 μM S107, which stabilizes the closed state of the ryanodine receptor 2, significantly decreased the percentage of CPVT-hiPSC-CMs presenting DADs to 25% (p<0.05).
CONCLUSIONS
We recapitulated the electrophysiological features of CPVT-derived hiPSC-CMs using electrical pacing. The development of DADs in the presence of isoproterenol was significantly suppressed by S107. Our model provides a promising platform to study disease mechanisms and screen drugs.
引言
人类诱导多能干细胞（hiPSC）为疾病建模提供了独特的机会。然而，由于hiPSC衍生的心肌细胞（hiPSC-CM）不成熟，重现疾病表型并非总是成功的。本研究的目的是建立并分析基于诱导多能干细胞的儿茶酚胺能多形性室性心动过速（CPVT）模型，该模型以肾上腺素能介导的致死性心律失常为特征，更确切地说是使用电起搏来促进新药物疗法的开发。
方法与结果
我们从一名37岁的CPVT患者身上获取hiPSC，并将其分化为心肌细胞。在自发搏动条件下，对照hiPSC-CM和CPVT-hiPSC-CM之间自发Ca2+瞬变的时间不规则性没有显著差异。在1Hz电场刺激下使用Ca2+成像，异丙肾上腺素在CPVT-hiPSC-CM中比在对照hiPSC-CM中更频繁地诱导异常舒张期Ca2+增加（对照12%对CPVT 43%，p<0.05）。自发搏动的hiPSC-CM的动作电位记录显示，对照细胞和CPVT细胞之间延迟后去极化（DAD）的频率没有显著差异。在1Hz起搏下应用异丙肾上腺素后，87.5%的CPVT-hiPSC-CM出现DAD，而对照hiPSC-CM为30%（p<0.05）。用10μM S107预孵育，可稳定兰尼碱受体2的关闭状态，使出现DAD的CPVT-hiPSC-CM百分比显著降低至25%（p<0.05）。
结论
我们使用电起搏重现了CPVT衍生的hiPSC-CM的电生理特征。S107显著抑制了异丙肾上腺素存在下DAD的发生。我们的模型为研究疾病机制和筛选药物提供了一个有前景的平台。
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