Patient-Specific and Genome-Edited Induced Pluripotent Stem Cell-Derived Cardiomyocytes Elucidate Single-Cell Phenotype of Brugada Syndrome

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• 作者：Ping Liang ; MD ; PhD^a ; ^b ; ^c ; ^{pingliang@zju.edu.cn" class="auth_mail" title="E-mail the corresponding author} ; Karim Sallam ; MD^a ; Haodi Wu ; PhD^a ; Yingxin Li ; PhD^a ; Ilanit Itzhaki ; PhD^a ; Priyanka Garg ; PhD^a ; Ying Zhang ; PhD^a ; Vittavat Termglichan ; MD^a ; Feng Lan ; PhD^a ; Mingxia Gu ; PhD^a ; Tingyu Gong ; MD^a ; ^b ; Yan Zhuge ; PhD^a ; Chunjiang He ; PhD^a ; Antje D. Ebert ; PhD^a ; Veronica Sanchez-Freire ; PhD^a ; Jared Churko ; PhD^a ; Shijun Hu ; PhD^a ; Arun Sharma ; PhD^a ; Chi Keung Lam ; PhD^a ; Melvin M. Scheinman ; MD^d ; Donald M. Bers ; PhD^e ; Joseph C. Wu ; MD ; PhD^a ; ^{joewu@stanford.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：action potential ; arrhythmia ; Ca2+ transient ; gene expression ; genome editing ; SCN5A
• 刊名：Journal of the American College of Cardiology
• 出版年：2016
• 出版时间：8 November 2016
• 年：2016
• 卷：68
• 期：19
• 页码：2086-2096
• 全文大小：2129 K

文摘

Brugada syndrome (BrS), a disorder associated with characteristic electrocardiogram precordial ST-segment elevation, predisposes afflicted patients to ventricular fibrillation and sudden cardiac death. Despite marked achievements in outlining the organ level pathophysiology of the disorder, the understanding of human cellular phenotype has lagged due to a lack of adequate human cellular models of the disorder.

Objectives

The objective of this study was to examine single cell mechanism of Brugada syndrome using induced pluripotent stem cell–derived cardiomyocytes (iPSC-CMs).

Methods

This study recruited 2 patients with type 1 BrS carrying 2 different sodium voltage-gated channel alpha subunit 5 variants as well as 2 healthy control subjects. We generated iPSCs from their skin fibroblasts by using integration-free Sendai virus. We used directed differentiation to create purified populations of iPSC-CMs.

Results

BrS iPSC-CMs showed reductions in inward sodium current density and reduced maximal upstroke velocity of action potential compared with healthy control iPSC-CMs. Furthermore, BrS iPSC-CMs demonstrated increased burden of triggered activity, abnormal calcium (Ca²⁺) transients, and beating interval variation. Correction of the causative variant by genome editing was performed, and resultant iPSC-CMs showed resolution of triggered activity and abnormal Ca²⁺ transients. Gene expression profiling of iPSC-CMs showed clustering of BrS compared with control subjects. Furthermore, BrS iPSC-CM gene expression correlated with gene expression from BrS human cardiac tissue gene expression.

Conclusions

Patient-specific iPSC-CMs were able to recapitulate single-cell phenotype features of BrS, including blunted inward sodium current, increased triggered activity, and abnormal Ca²⁺ handling. This novel human cellular model creates future opportunities to further elucidate the cellular disease mechanism and identify novel therapeutic targets.