Genome editing of isogenic human induced pluripotent stem cells recapitulates long QT phenotype for drug testing

Yongming Wang; Ping Liang; Feng Lan; Haodi Wu; Leszek Lisowski; Mingxia Gu; Shijun Hu; Mark A Kay; Fyodor D Urnov; Rami Shinnawi; Joseph D Gold; Lior Gepstein; Joseph C Wu

doi:10.1016/j.jacc.2014.04.057

Genome editing of isogenic human induced pluripotent stem cells recapitulates long QT phenotype for drug testing

J Am Coll Cardiol. 2014 Aug 5;64(5):451-9. doi: 10.1016/j.jacc.2014.04.057.

Authors

Yongming Wang¹, Ping Liang², Feng Lan², Haodi Wu², Leszek Lisowski³, Mingxia Gu², Shijun Hu², Mark A Kay³, Fyodor D Urnov⁴, Rami Shinnawi⁵, Joseph D Gold⁶, Lior Gepstein⁵, Joseph C Wu⁷

Affiliations

¹ Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, China.
² Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California.
³ Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, California.
⁴ Sangamo Biosciences, Richmond, California.
⁵ Rambam Medical Center, Technion, Israel Institute of Technology, Haifa, Israel.
⁶ Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California.
⁷ Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, California; Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California; Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California. Electronic address: joewu@stanford.edu.

Abstract

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.

Keywords: disease models; drug testing; genome editing; long QT syndrome; stem cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Anti-Arrhythmia Agents / therapeutic use*
Cell Differentiation
Cell Transdifferentiation
Cells, Cultured
Genome
Humans
Induced Pluripotent Stem Cells / metabolism
Induced Pluripotent Stem Cells / pathology*
Long QT Syndrome / drug therapy
Long QT Syndrome / genetics*
Long QT Syndrome / pathology
Patch-Clamp Techniques

Substances

Anti-Arrhythmia Agents

Abstract

Publication types

MeSH terms

Substances

Grants and funding