Patient-Specific Human Induced Pluripotent Stem Cell Model Assessed with Electrical Pacing Validates S107 as a Potential Therapeutic Agent for Catecholaminergic Polymorphic Ventricular Tachycardia

PLOS ONE, Dec 2019

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.

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Patient-Specific Human Induced Pluripotent Stem Cell Model Assessed with Electrical Pacing Validates S107 as a Potential Therapeutic Agent for Catecholaminergic Polymorphic Ventricular Tachycardia

October Patient-Specific Human Induced Pluripotent Stem Cell Model Assessed with Electrical Pacing Validates S107 as a Potential Therapeutic Agent for Catecholaminergic Polymorphic Ventricular Tachycardia Kenichi Sasaki 1 2 Takeru Makiyama 1 2 Yoshinori Yoshida 0 2 Yimin Wuriyanghai 1 2 Tsukasa Kamakura 1 2 Suguru Nishiuchi 1 2 Mamoru Hayano 1 2 Takeshi Harita 1 2 Yuta Yamamoto 1 2 Hirohiko Kohjitani 1 2 Sayako Hirose 1 2 Jiarong Chen 1 2 Mihoko Kawamura 2 Seiko Ohno 2 Hideki Itoh 2 Ayako Takeuchi 2 Satoshi Matsuoka 2 Masaru Miura 2 Naokata Sumitomo 2 Minoru Horie 2 Shinya Yamanaka 0 2 Takeshi Kimura 1 2 0 Kyoto University iPS Cell Research and Application, Kyoto, Japan, 3 Department of Cardiovascular and Respiratory Medicine, Shiga University of Medical Science , Otsu , Japan , 4 Department of Integrative and Systems Physiology, Faculty of Medical Sciences, University of Fukui, Fukui, Japan, 5 Division of Cardiology, Tokyo Metropolitan Children's Medical Center, Tokyo, Japan, 6 Department of Pediatric Cardiology, Saitama Medical University International Medical Center , Saitama , Japan 1 Department of Cardiovascular Medicine, Kyoto University Graduate School of Medicine , Kyoto , Japan 2 Editor: Tomohiko Ai, Indiana University , UNITED STATES 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. - OPEN ACCESS Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: This work was supported by Japan Society for the Promotion of Science KAKENHI Grant Number 25461054 and Suzuken Memorial Foundation Grant Number 10-097. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Method and Results We generated hiPSCs from a 37-year-old CPVT patient and differentiated them into cardio myocytes. 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 Competing Interests: The authors have declared that no competing interests exist. 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 rya nodine 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. Introduction Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a hereditary arrhythmic disorder characterized by bidirectional ventricular tachycardia (VT) that is triggered by emotional stress or physical exercise and leads to syncope or sudden cardiac death without structural heart disease. CPVT is caused by autosomal dominant mutations in the cardiac ryanodine receptor gene (RyR2) [ 1 ], the calmodulin gene (CALM1, CALM2) [ 2, 3 ] and the inward rectifying potassium channel gene (KCNJ2) [4] and autosomal recessive mutations in the cardiac calsequestrin gene (CASQ2) [ 5 ] and the triadin gene (TRDN) [ 6 ]. Approximately 50–55% of CPVT cases are associated with RyR2 mutations [ 7 ] and 1–2% are due to CASQ2 mutations [ 8 ]. Beta-blockers are the first-line therapy for CPVT, but they often fail to prevent fatal arrhythmias [ 9 ]. Recently, flecainide, a class Ic Na+ channel blocker, has been reported to be effective for treating CPVT patients [ 10 ]. The advent of human induced pluripotent stem cell (hiPSC) technology has enabled us to use human cardiomyocytes that have the same genetic background as the patients. There have been several reports of CPVT iPSC-based model [ 11–15 ], however, it is becoming clearer that hiPSC-derived cardiomyocytes (hiPSC-CMs) have immature electrophysiological and structural properties compared t (...truncated)


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Kenichi Sasaki, Takeru Makiyama, Yoshinori Yoshida, Yimin Wuriyanghai, Tsukasa Kamakura, Suguru Nishiuchi, Mamoru Hayano, Takeshi Harita, Yuta Yamamoto, Hirohiko Kohjitani, Sayako Hirose, Jiarong Chen, Mihoko Kawamura, Seiko Ohno, Hideki Itoh, Ayako Takeuchi, Satoshi Matsuoka, Masaru Miura, Naokata Sumitomo, Minoru Horie, Shinya Yamanaka, Takeshi Kimura. Patient-Specific Human Induced Pluripotent Stem Cell Model Assessed with Electrical Pacing Validates S107 as a Potential Therapeutic Agent for Catecholaminergic Polymorphic Ventricular Tachycardia, PLOS ONE, 2016, Volume 11, Issue 10, DOI: 10.1371/journal.pone.0164795