4.4 Article

CRISPR correction of the PRKAG2 gene mutation in the patient's induced pluripotent stem cell-derived cardiomyocytes eliminates electrophysiological and structural abnormalities

期刊

HEART RHYTHM
卷 15, 期 2, 页码 267-276

出版社

ELSEVIER SCIENCE INC
DOI: 10.1016/j.hrthm.2017.09.024

关键词

arrhythmia; clustered regularly interspaced short palindromic repeats; electrophysiology; induced pluripotent stem cell-derived cardiomyocyte; PRKAG2; Wolff-Parkinson-White syndrome

资金

  1. Israel Science Foundation (ISF)
  2. US-Israel BSF
  3. Israeli Ministry of Science and Technology
  4. Rappaport Institute, Niedersachsisches Ministerium: Medizinischen Hochschule Hannover (MHH) [11-76251-99-16/14]
  5. German Research Foundation [574157]
  6. DZHK [B15-006 SE]

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BACKGROUND Mutations in the PRKAG2 gene encoding the gamma-sub-unit of adenosine monophosphate kinase (AMPK) cause hypertrophic cardiomyopathy (HCM) and familial Wolff-Parkinson-White (WPW) syndrome. Patients carrying the R302Q mutation in PRKAG2 present with sinus bradycardia, escape rhythms, ventricular preexcitation, supraventricular tachycardia, and atrioventricular block. This mutation affects AMPK activity and increases glycogen storage in cardiomyocytes. The link between glycogen storage, WPW syndrome, HCM, and arrhythmias remains unknown. OBJECTIVE The purpose of this study was to investigate the pathological changes caused by the PRKAG2 mutation. We tested the hypothesis that patient's induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) display clinical aspects of the disease. METHODS Using clustered regularly interspaced short palindromic repeats (CRISPR) technology, we corrected the mutation and then generated isogenic iPSC-CMs. Action potentials were recorded from spontaneously firing and paced cardiomyocytes using the patch clamp technique. Using a microelectrode array setup, we recorded electrograms from iPSC-CMs clusters. Transmission electron microscopy was used to detect ultrastructural abnormalities in the mutated iPSC-CMs. RESULTS PRKAG2-mutated iPSC-CMs exhibited abnormal firing patterns, delayed after depolarizations, triggered arrhythmias, and augmented beat rate variability. Importantly, CRISPR correction eliminated the electrophysiological abnormalities, the augmented glycogen, storage, and cardiomyocyte hypertrophy. CONCLUSION PRKAG2-mutated iPSC-CMs displayed functional and structural abnormalities, which were abolished by correcting the mutation in the patient's iPSCs using CRISPR technology.

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