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

Ronen Ben Jehuda, Binyamin Eisen, Yuval Shemer, Lucy N. Mekies, Agnes Szantai, Irina Reiter, Huanhuan Cui, Kaomei Guan, Shiraz Haron-Khun, Dov Freimark, Silke R. Sperling, Mihaela Gherghiceanu, Michael Arad, Ofer Binah*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Background Mutations in the PRKAG2 gene encoding the γ-subunit 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 afterdepolarizations, 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.

Original languageEnglish
Pages (from-to)267-276
Number of pages10
JournalHeart Rhythm
Volume15
Issue number2
DOIs
StatePublished - Feb 2018

Funding

FundersFunder number
Niedersachsisches Ministerium: Medizinischen Hochschule Hannover
Rappaport Institute
US-Israel BSF
Deutsches Zentrum für Herz-KreislaufforschungB15-006 SE
Deutsches Zentrum für Herz-Kreislaufforschung
Maison Heinrich Heine11-76251-99-16/14
Maison Heinrich Heine
Deutsche Forschungsgemeinschaft574157
Deutsche Forschungsgemeinschaft
Israel Science Foundation
Ministry of Science and Technology, Israel

    Keywords

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

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