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


Karakikes I, Stillitano F, Nonnenmacher M, Tzimas C, Sanoudou D, Termglinchan V, Kong CW, Rushing S, Hansen J, Ceholski D, et al. Correction of human phospholamban R14del mutation associated with cardiomyopathy using targeted nucleases and combination therapy. Nat CommunNat CommunNat Commun. 2015;6:6955.


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.


Karakikes, IoannisStillitano, FrancescaNonnenmacher, MathieuTzimas, ChristosSanoudou, DespinaTermglinchan, VittavatKong, Chi-WingRushing, StephanieHansen, JensCeholski, DelaineKolokathis, FotisKremastinos, DimitriosKatoulis, AlexandrosRen, LihuanCohen, NinetteGho, Johannes M I HTsiapras, DimitriosVink, AryanWu, Joseph CAsselbergs, Folkert WLi, Ronald AHulot, Jean-SebastienKranias, Evangelia GHajjar, Roger JengP50 HL112324/HL/NHLBI NIH HHS/R01 HL119046/HL/NHLBI NIH HHS/R01 HL123968/HL/NHLBI NIH HHS/T32 HL007824/HL/NHLBI NIH HHS/R00 HL104002/HL/NHLBI NIH HHS/15BGIA22730027/AHA/American Heart Association-American Stroke Association/R01 HL126527/HL/NHLBI NIH HHS/R01 HL064018/HL/NHLBI NIH HHS/R01 HL093183/HL/NHLBI NIH HHS/HL26057/HL/NHLBI NIH HHS/R37 HL026057/HL/NHLBI NIH HHS/HHSN268201000045C/HL/NHLBI NIH HHS/R01 HL117505/HL/NHLBI NIH HHS/R01 HL026057/HL/NHLBI NIH HHS/K99 HL104002/HL/NHLBI NIH HHS/HL 119046/HL/NHLBI NIH HHS/Research Support, N.I.H., ExtramuralResearch Support, Non-U.S. Gov'tEngland2015/04/30 06:00Nat Commun. 2015 Apr 29;6:6955. doi: 10.1038/ncomms7955.