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Nat Commun. 2015 Apr 29;6:6955. doi: 10.1038/ncomms7955.

Correction of human phospholamban R14del mutation associated with cardiomyopathy using targeted nucleases and combination therapy.

Author information

1
1] Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA [2] Department of Medicine, Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, California 94305, USA [3] Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California 94305, USA.
2
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
3
Molecular Biology, Biomedical Research Foundation of the Academy of Athens, Athens, Greece.
4
Pharmacology Department, University of Athens, Athens, Greece.
5
1] Department of Medicine, Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, California 94305, USA [2] Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California 94305, USA.
6
Stem Cell &Regenerative Medicine Consortium, Department of Physiology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, China.
7
Department of Pharmacology and Systems Therapeutics, Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
8
Attikon Hospital, University of Athens, Athens 12462, Greece.
9
Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
10
Department of Cardiology, Division Heart and Lungs, University Medical Centre Utrecht, PO Box 85500, 3508 GA, Utrecht, The Netherlands.
11
Ona ssis Cardiac Surgery Center, Athens, Greece.
12
Department of Pathology, Division Laboratories and Pharmacy, University Medical Center Utrecht, PO Box 85500, 3508 GA, Utrecht, The Netherlands.
13
1] Department of Cardiology, Division Heart and Lungs, University Medical Centre Utrecht, PO Box 85500, 3508 GA, Utrecht, The Netherlands [2] Durrer Center for Cardiogenetic Research, ICIN-Netherlands Heart Institute, Utrecht, The Netherlands [3] Institute of Cardiovascular Science, faculty of Population Health Sciences, University College London, London WC1E 6BT, UK.
14
1] Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA [2] Stem Cell &Regenerative Medicine Consortium, Department of Physiology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, China.
15
1] Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA [2] Sorbonne Universités, UPMC Univ Paris 06, INSERM, UMR_S 1166 ICAN, F-75005 Paris, France.
16
1] Molecular Biology, Biomedical Research Foundation of the Academy of Athens, Athens, Greece [2] Department of Pharmacology, University of Cincinnati, Cincinnati, Ohio 45267-0575, USA.

Abstract

A number of genetic mutations is associated with cardiomyopathies. A mutation in the coding region of the phospholamban (PLN) gene (R14del) is identified in families with hereditary heart failure. Heterozygous patients exhibit left ventricular dilation and ventricular arrhythmias. Here we generate induced pluripotent stem cells (iPSCs) from a patient harbouring the PLN R14del mutation and differentiate them into cardiomyocytes (iPSC-CMs). We find that the PLN R14del mutation induces Ca(2+) handling abnormalities, electrical instability, abnormal cytoplasmic distribution of PLN protein and increases expression of molecular markers of cardiac hypertrophy in iPSC-CMs. Gene correction using transcription activator-like effector nucleases (TALENs) ameliorates the R14del-associated disease phenotypes in iPSC-CMs. In addition, we show that knocking down the endogenous PLN and simultaneously expressing a codon-optimized PLN gene reverses the disease phenotype in vitro. Our findings offer novel strategies for targeting the pathogenic mutations associated with cardiomyopathies.

PMID:
25923014
PMCID:
PMC4421839
DOI:
10.1038/ncomms7955
[Indexed for MEDLINE]
Free PMC Article

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