TY - JOUR
T1 - Correction of human phospholamban R14del mutation associated with cardiomyopathy using targeted nucleases and combination therapy
AU - Karakikes, Ioannis
AU - Stillitano, Francesca
AU - Nonnenmacher, Mathieu
AU - Tzimas, Christos
AU - Sanoudou, Despina
AU - Termglinchan, Vittavat
AU - Kong, Chi-Wing
AU - Rushing, Stephanie
AU - Hansen, Jens
AU - Ceholski, Delaine
AU - Kolokathis, Fotis
AU - Kremastinos, Dimitrios
AU - Katoulis, Alexandros
AU - Ren, Lihuan
AU - Cohen, Ninette
AU - Gho, Johannes M. I. H.
AU - Tsiapras, Dimitrios
AU - Vink, Aryan
AU - Wu, Joseph C.
AU - Asselbergs, Folkert W.
AU - Li, Ronald A.
AU - Hulot, Jean-Sebastien
AU - Kranias, Evangelia G.
AU - Hajjar, Roger J.
PY - 2015/4/29
Y1 - 2015/4/29
N2 - 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.
AB - 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.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84928966984&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/25923014
U2 - https://doi.org/10.1038/ncomms7955
DO - https://doi.org/10.1038/ncomms7955
M3 - Article
C2 - 25923014
SN - 2041-1723
VL - 6
JO - Nature communications
JF - Nature communications
M1 - 6955
ER -