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

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.

• Authors: Ioannis Karakikes, Francesca Stillitano, Mathieu Nonnenmacher, Christos Tzimas, Despina Sanoudou, Vittavat Termglinchan, Chi-Wing Kong, Stephanie Rushing, Jens Hansen, Delaine Ceholski, Fotis Kolokathis, Dimitrios Kremastinos, Alexandros Katoulis, Lihuan Ren, Ninette Cohen, Johannes M I H Gho, Dimitrios Tsiapras, Aryan Vink, Joseph C Wu, Folkert W Asselbergs, Ronald A Li, Jean-Sebastien Hulot, Evangelia G Kranias, Roger J Hajjar
• Journal: Nature communications (Nat Commun) Vol. 6 Pg. 6955 (Apr 29 2015) ISSN: 2041-1723 [Electronic] England
• PMID: 25923014 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Calcium-Binding Proteins
• phospholamban
• Deoxyribonucleases

Topics

• Adenoviridae
• Adult
• Calcium-Binding Proteins (genetics)
• Cardiomyopathies (genetics, metabolism, therapy)
• Deoxyribonucleases
• Female
• Gene Transfer Techniques
• Humans
• Induced Pluripotent Stem Cells
• Myocytes, Cardiac (metabolism)
• Phenotype
• Sequence Deletion
• Targeted Gene Repair