Establishment of a PRKAG2 cardiac syndrome disease model and mechanism study using human induced pluripotent stem cells

J Mol Cell Cardiol. 2018 Apr:117:49-61. doi: 10.1016/j.yjmcc.2018.02.007. Epub 2018 Feb 13.

Abstract

PRKAG2 cardiac syndrome is a distinct form of human cardiomyopathy characterized by cardiac hypertrophy, ventricular pre-excitation and progressive cardiac conduction disorder. However, it remains unclear how mutations in the PRKAG2 gene give rise to such a complicated disease. To investigate the underlying molecular mechanisms, we generated disease-specific hiPSC-derived cardiomyocytes from two brothers both carrying a heterozygous missense mutation c.905G>A (R302Q) in the PRKAG2 gene and further corrected the R302Q mutation with CRISPR-Cas9 mediated genome editing. Disease-specific hiPSC-cardiomyocytes recapitulated many phenotypes of PRKAG2 cardiac syndrome including cellular enlargement, electrophysiological irregularities and glycogen storage. In addition, we found that the PRKAG2-R302Q mutation led to increased AMPK activities, resulting in extensive glycogen deposition and cardiomyocyte hypertrophy. Finally we confirmed that disrupted phenotypes of PRKAG2 cardiac syndrome caused by the specific PRKAG2-R302Q mutation can be alleviated by small molecules inhibiting AMPK activity and be rescued with CRISPR-Cas9 mediated genome correction. Our results showed that disease-specific hiPSC-CMs and genetically-corrected hiPSC-cardiomyocytes would be a very useful platform for understanding the pathogenesis of, and testing autologous cell-based therapies for, PRKAG2 cardiac syndrome.

Keywords: AMPK activity; Cardiac hypertrophy; Cardiomyocytes; Glycogen storage; Human induced pluripotent stem cells; PRKAG2 cardiac syndrome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AMP-Activated Protein Kinases / antagonists & inhibitors
  • AMP-Activated Protein Kinases / genetics
  • AMP-Activated Protein Kinases / metabolism*
  • Adult
  • Base Sequence
  • Calcium / metabolism
  • Cardiomegaly / enzymology
  • Cardiomegaly / pathology
  • Cell Differentiation
  • Electrophysiological Phenomena
  • Glycogen / metabolism
  • Heart Diseases / enzymology*
  • Heart Diseases / pathology*
  • Heart Diseases / physiopathology
  • Humans
  • Induced Pluripotent Stem Cells / metabolism*
  • Male
  • Mitochondria / metabolism
  • Models, Biological*
  • Mutation / genetics
  • Myocardial Contraction
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / pathology
  • Myocytes, Cardiac / ultrastructure
  • Oxidation-Reduction
  • Phenotype
  • Reproducibility of Results
  • Syndrome

Substances

  • Glycogen
  • PRKAG2 protein, human
  • AMP-Activated Protein Kinases
  • Calcium