Abstract
Dilated cardiomyopathy (DCM) can be caused by mutations in the cardiac protein phospholamban (PLN). We used CRISPR/Cas9 to insert the R9C PLN mutation at its endogenous locus into a human induced pluripotent stem cell (hiPSC) line from an individual with no cardiovascular disease. R9C PLN hiPSC-CMs display a blunted β-agonist response and defective calcium handling. In 3D human engineered cardiac tissues (hECTs), a blunted lusitropic response to β-adrenergic stimulation was observed with R9C PLN. hiPSC-CMs harboring the R9C PLN mutation showed activation of a hypertrophic phenotype, as evidenced by expression of hypertrophic markers and increased cell size and capacitance of cardiomyocytes. RNA-seq suggests that R9C PLN results in an altered metabolic state and profibrotic signaling, which was confirmed by gene expression analysis and picrosirius staining of R9C PLN hECTs. The expression of several miRNAs involved in fibrosis, hypertrophy, and cardiac metabolism were also perturbed in R9C PLN hiPSC-CMs. This study contributes to better understanding of the pathogenic mechanisms of the hereditary R9C PLN mutation in the context of human cardiomyocytes.
Original language | English (US) |
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Pages (from-to) | 147-154 |
Number of pages | 8 |
Journal | Journal of Molecular and Cellular Cardiology |
Volume | 119 |
DOIs | |
State | Published - Jun 2018 |
Bibliographical note
Funding Information:This work is supported by the National Institutes of Health ( R01 HL117505 , HL 132226 , HL119046 , HL129814 , HL128072 , HL128099 , HL132684 , HL131404 , and HL135093 ; and R37 AG026160 ), and a Transatlantic Fondation Leducq grant (Cellular and Molecular Targets to Promote Therapeutic Cardiac Regeneration). DKC was a fellow of the American Heart Association (15POST25090116). ICT is supported by NIH / NHLBI K01 HL 133424-01 . We would like to acknowledge the Microscopy Core and Black Family Stem Cell Institute at the Icahn School of Medicine at Mount Sinai.
Funding Information:
This work is supported by the National Institutes of Health (R01 HL117505, HL 132226, HL119046, HL129814, HL128072, HL128099, HL132684, HL131404, and HL135093; and R37 AG026160), and a Transatlantic Fondation Leducq grant (Cellular and Molecular Targets to Promote Therapeutic Cardiac Regeneration). DKC was a fellow of the American Heart Association (15POST25090116). ICT is supported by NIH/NHLBIK01 HL 133424-01. We would like to acknowledge the Microscopy Core and Black Family Stem Cell Institute at the Icahn School of Medicine at Mount Sinai.
Publisher Copyright:
© 2018
Keywords
- CRISPR/Cas9
- Cardiomyocytes
- Dilated cardiomyopathy
- Engineered cardiac tissue
- Human induced pluripotent stem cells
- Phospholamban