Pathogenic mechanism and gene correction for LQTS-causing double mutations in KCNQ1 using a pluripotent stem cell model

Zhen Wang; Lipeng Wang; Wenling Liu; Dayi Hu; Yuanfeng Gao; Qing Ge; Xin Liu; Lei Li; Yangming Wang; Shiqiang Wang; Cuilan Li

doi:10.1016/j.scr.2019.101483

Pathogenic mechanism and gene correction for LQTS-causing double mutations in KCNQ1 using a pluripotent stem cell model

Stem Cell Res. 2019 Jul:38:101483. doi: 10.1016/j.scr.2019.101483. Epub 2019 Jun 11.

Authors

Zhen Wang¹, Lipeng Wang², Wenling Liu¹, Dayi Hu¹, Yuanfeng Gao¹, Qing Ge¹, Xin Liu¹, Lei Li¹, Yangming Wang³, Shiqiang Wang⁴, Cuilan Li⁵

Affiliations

¹ Heart Center, Peking University People's Hospital, Beijing, China.
² State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University, Beijing, China.
³ Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Institute of Molecular Medicine, Peking University, Beijing, China. Electronic address: yangming.wang@pku.edu.cn.
⁴ State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University, Beijing, China. Electronic address: wsq@pku.edu.cn.
⁵ Heart Center, Peking University People's Hospital, Beijing, China. Electronic address: licuilan@pkuph.edu.cn.

PMID: 31226583
DOI: 10.1016/j.scr.2019.101483

Abstract

Aims: To establish a KCNQ1 mutant-specific induced pluripotent stem cell (iPSC) model of a Chinese inherited long QT syndrome (LQTS) patient and to explore the pathogenesis of KCNQ1 mutations.

Methods and results: (1) Two patient-specific iPSC lines from the proband were obtained. (2) The experiments produced spontaneously beating cardiomyocytes (CMs) from patient iPSCs. Splicing mutation c. 605-2A > G in iPSC-derived cardiomyocytes (iPSC-CMs) resulted in the skipping of exon 4, exons 3-4, or exons 3-6 in KCNQ1 transcription what was observed in the patient's peripheral leukocytes. (3) Action potential duration (APD) at 50% and 90% repolarization (APD50 and APD90) of the patient's iPSC-derived ventricular-like-CMs was significantly longer than that of the control. Moreover, early after depolarization (EAD) and coupled beats were observed only in L1-iPSC-CMs. (4) A c.815G > A corrected iPSC line was obtained by using the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein 9 (Cas9) system.

Conclusion: (1) Cardiomyocytes with spontaneous pulsation were successfully differentiated from LQTS patient-specific iPSC lines. (2) For KCNQ1 splicing mutations, there is a chance that splicing patterns in peripheral leukocytes are similar to that in patient iPSC-CMs. (3) The truncated KCNQ1 proteins induced by such splicing mutation might cause Iks decrease, which in turn produced APD prolongation and triggered activities. (4) Our data showed that CRISPR-Cas9 system could be used to rescue the LQTS-related mutations.

Keywords: Crispr-Cas9; KCNQ1; LQTS; Splicing mutation; iPSC.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems*
Gene Editing*
Humans
Induced Pluripotent Stem Cells* / metabolism
Induced Pluripotent Stem Cells* / pathology
KCNQ1 Potassium Channel*
Long QT Syndrome* / genetics
Long QT Syndrome* / metabolism
Long QT Syndrome* / pathology
Long QT Syndrome* / therapy
Models, Cardiovascular*
Mutation*
Myocytes, Cardiac / metabolism
Myocytes, Cardiac / pathology

Substances

KCNQ1 Potassium Channel
KCNQ1 protein, human