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

• DOI: 10.1038/ncomms7955
• Publication Date: 2015-04-29T13:25:34Z
• Authors: Ioannis Karakikes; Francesca Stillitano; Mathieu Nonnenmacher; Christos Tzimas; Despina Sanoudou; Vittavat Termglin...; Chi-Wing Kong; Stephanie Rushing; Jens Hansen; Delaine Ceholski; Fotis Kolokathis; Dimitrios Kremast...; Alexandros Katoulis; Lihuan Ren; Ninette Cohen; Johannes M.I.H. Gho; Dimitrios Tsiapras; Aryan Vink; Joseph C. Wu; Folkert W. Asselb...; Ronald A. Li; Jean-Sebastien Hulot; Evangelia G. Kranias; Roger J. Hajjar
• Keywords: Adult; Induced Pluripotent Stem Cells; LONG-QT SYNDROME; ARRHYTHMOGENIC CARDIOMYOPATHY; CARDIOMYOCYTES; DISEASE; Article; Adenoviridae; 03 medical and health sciences; Humans; Myocytes, Cardiac; Sequence Deletion; 0303 health sciences; [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology; Deoxyribonucleases; HYPERTROPHIC CARDIOMYOPATHY; DIRECTED DIFFERENTIATION; Calcium-Binding Proteins; Gene Transfer Techniques; IN-VITRO; DILATED CARDIOMYOPATHY; 3. Good health; Phenotype; Female; Cardiomyopathies; PLURIPOTENT STEM-CELLS; [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology; ADENOASSOCIATED VIRUS VECTORS; Targeted Gene Repair

AbstractA 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 Ca2+ 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.