Correction of human phospholamban R14del mutation associated with cardiomyopathy using targeted nucleases and combination therapy.
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.
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.|Department of Medicine, Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, California 94305, USA.|Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California 94305, USA.
Ioannis
I
Karakikes
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
Francesca
F
Stillitano
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
Mathieu
M
Nonnenmacher
Molecular Biology, Biomedical Research Foundation of the Academy of Athens, Athens, Greece.
Christos
C
Tzimas
Pharmacology Department, University of Athens, Athens, Greece.
Despina
http://orcid.org/0000-0003-3704-1941
D
Sanoudou
Department of Medicine, Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, California 94305, USA.|Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California 94305, USA.
Vittavat
V
Termglinchan
Stem Cell &Regenerative Medicine Consortium, Department of Physiology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, China.
Chi-Wing
CW
Kong
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
Stephanie
S
Rushing
Department of Pharmacology and Systems Therapeutics, Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
Jens
J
Hansen
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
Delaine
D
Ceholski
Attikon Hospital, University of Athens, Athens 12462, Greece.
Fotis
F
Kolokathis
Attikon Hospital, University of Athens, Athens 12462, Greece.
Dimitrios
D
Kremastinos
Attikon Hospital, University of Athens, Athens 12462, Greece.
Alexandros
A
Katoulis
Stem Cell &Regenerative Medicine Consortium, Department of Physiology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, China.
Lihuan
L
Ren
Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
Ninette
N
Cohen
Department of Cardiology, Division Heart and Lungs, University Medical Centre Utrecht, PO Box 85500, 3508 GA, Utrecht, The Netherlands.
Johannes M I H
JMIH
Gho
Ona ssis Cardiac Surgery Center, Athens, Greece.
Dimitrios
D
Tsiapras
Department of Pathology, Division Laboratories and Pharmacy, University Medical Center Utrecht, PO Box 85500, 3508 GA, Utrecht, The Netherlands.
Aryan
A
Vink
Department of Medicine, Division of Cardiovascular Medicine, Stanford University School of Medicine, Stanford, California 94305, USA.|Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, California 94305, USA.
Joseph C
JC
Wu
Department of Cardiology, Division Heart and Lungs, University Medical Centre Utrecht, PO Box 85500, 3508 GA, Utrecht, The Netherlands.|Durrer Center for Cardiogenetic Research, ICIN-Netherlands Heart Institute, Utrecht, The Netherlands.|Institute of Cardiovascular Science, faculty of Population Health Sciences, University College London, London WC1E 6BT, UK.
Folkert W
http://orcid.org/0000-0002-1692-8669
FW
Asselbergs
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.|Stem Cell &Regenerative Medicine Consortium, Department of Physiology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, China.
Ronald A
RA
Li
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.|Sorbonne Universités, UPMC Univ Paris 06, INSERM, UMR_S 1166 ICAN, F-75005 Paris, France.
Jean-Sebastien
JS
Hulot
Molecular Biology, Biomedical Research Foundation of the Academy of Athens, Athens, Greece.|Department of Pharmacology, University of Cincinnati, Cincinnati, Ohio 45267-0575, USA.
Evangelia G
EG
Kranias
Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
Roger J
RJ
Hajjar
D000256:Adenoviridae; D000328:Adult; D002135:Calcium-Binding Proteins / Q000235:genetics*; D009202:Cardiomyopathies / Q000235:genetics* / Q000378:metabolism / Q000628:therapy; D003851:Deoxyribonucleases; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D032383:Myocytes, Cardiac / Q000378:metabolism*; D010641:Phenotype; D017384:Sequence Deletion; D052416:Targeted Gene Repair; D000099211:Phospholamban
D002135:Calcium-Binding Proteins; D003851:Deoxyribonucleases; D000099211:Phospholamban