Genome Editing of Isogenic Human Induced Pluripotent Stem Cells Recapitulates Long QT Phenotype for Drug Testing

JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY

卷:64

Article

Wang, Yongming^1,2,3  Liang, Ping^1,2,4  Lan, Feng^1,2,4  Wu, Haodi^1,2,4  Lisowski, Leszek^5,6  Gu, Mingxia^1,2,4  Hu, Shijun^1,2,4  Kay, Mark A.^5,6  Urnov, Fyodor D.⁷  Shinnawi, Rami⁸  Gold, Joseph D.^1,2  Gepstein, Lior⁸  Wu, Joseph C.^1,2,4 

[1] Stanford Univ, Dept Med, Div Cardiol, Sch Med, Stanford, CA 94305 USA

[2] Stanford Univ, Sch Med, Stanford Cardiovasc Inst, Stanford, CA USA

[3] Fudan Univ, State Key Lab Genet Engn, Shanghai 200433, Peoples R China

[4] Stanford Univ, Sch Med, Inst Stem Cell Biol & Regenerat Med, Stanford, CA USA

[5] Stanford Univ, Dept Pediat, Sch Med, Stanford, CA 94305 USA

[6] Stanford Univ, Dept Genet, Sch Med, Stanford, CA 94305 USA

[7] Sangamo Biosci, Richmond, CA USA

[8] Technion Israel Inst Technol, Rambam Med Ctr, Haifa, Israel

关键词: disease models;    drug testing;    genome editing;    long QT syndrome;    stem cells;   

DOI  :  10.1016/j.jacc.2014.04.057

来源: Elsevier

【 摘 要 】

BACKGROUND Human induced pluripotent stem cells (iPSCs) play an important role in disease modeling and drug testing. However, the current methods are time-consuming and lack an isogenic control. OBJECTIVES This study sought to establish an efficient technology to generate human PSC-based disease models with isogenic control. METHODS The ion channel genes KCNQ1 and KCNH2 with dominant negative mutations causing long QT syndrome types 1 and 2, respectively, were stably integrated into a safe harbor AAVS1 locus using zinc finger nuclease technology. RESULTS Patch-clamp recording revealed that the edited iPSC-derived cardiomyocytes (iPSC-CMs) displayed characteristic long QT syndrome phenotype and significant prolongation of the action potential duration compared with the unedited control cells. Finally, addition of nifedipine (L-type calcium channel blocker) or pinacidil (KATP-channel opener) shortened the action potential duration of iPSC-CMs, confirming the validity of isogenic iPSC lines for drug testing in the future. CONCLUSIONS Our study demonstrates that iPSC-CM-based disease models can be rapidly generated by overexpression of dominant negative gene mutants. (C) 2014 by the American College of Cardiology Foundation.

【 授权许可】

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