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Modeling the mitochondrial cardiomyopathy of Barth syndrome with induced pluripotent stem cell and heart-on-chip technologies



Study of monogenic mitochondrial cardiomyopathies may yield insights into mitochondrial roles in cardiac development and disease. Here, we combined patient-derived and genetically engineered induced pluripotent stem cells (iPSCs) with tissue engineering to elucidate the pathophysiology underlying the cardiomyopathy of Barth syndrome (BTHS), a mitochondrial disorder caused by mutation of the gene encoding tafazzin (TAZ). Using BTHS iPSC-derived cardiomyocytes (iPSC-CMs), we defined metabolic, structural and functional abnormalities associated with TAZ mutation. BTHS iPSC-CMs assembled sparse and irregular sarcomeres, and engineered BTHS 'heart-on-chip' tissues contracted weakly. Gene replacement and genome editing demonstrated that TAZ mutation is necessary and sufficient for these phenotypes. Sarcomere assembly and myocardial contraction abnormalities occurred in the context of normal whole-cell ATP levels. Excess levels of reactive oxygen species mechanistically linked TAZ mutation to impaired cardiomyocyte function. Our study provides new insights into the pathogenesis of Barth syndrome, suggests new treatment strategies and advances iPSC-based in vitro modeling of cardiomyopathy.

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Figure 1: Mitochondrial abnormalities in BTHS iPSC-CMs.
Figure 2: TAZ deficiency is necessary to cause the iPSC-CM metabolic phenotype.
Figure 3: Construction and characterization of TAZ mutant and isogenic control iPSCs by Cas9-mediated genome editing.
Figure 4: Sarcomere organization is impaired in BTH-H mutant iPSC-CMs.
Figure 5: BTHS myocardial tissue constructs exhibit depressed contractile stress generation.
Figure 6: Effect of small molecules on BTHS iPCS-CM ATP levels and mitochondrial function.

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This work supported by the Barth Syndrome Foundation, the Boston Children's Hospital Translational Investigator Service, the US National Institutes of Health (NIH) NHLBI Progenitor Cell Biology Consortium (NIH U01 HL100401 and U01 HL100408), NIH RC1 HL099618, NIH UH2 TR000522 and charitable donations from E. Marram, K. Carpenter and G.F. Smith.

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Authors and Affiliations



G.W. designed and performed experiments and analyzed data. M.L.M. designed and performed experiments on MTFs and sarcomere organization and analyzed data. L.Y. and G.M.C. provided expert assistance and reagents for genome editing. F.S.P. designed the sarcomere organization analysis method. H.Y. developed the MTF analysis method. A.A. assisted with MTF substrate fabrication and experiments. D.J. provided advice on mitochondrial assays. D.Z. imaged iPSC-CMs to assess their mitochondrial organization and potential. L.Z. and K.R.C. provided expert assistance with modRNA, and J.C., J.D. and D.-Z.W. helped construct modRNAs. K.L. contributed to genome editing. R.J.A.W., W.K. and F.M.V. analyzed phospholipids. M.A.L. and C.E.M. provided expert assistance in iPSC differentiation to cardiomyocytes. A.H. developed TAZ shRNA viruses and provided technical assistance. J.G. and A.E.R. obtained patient samples. Q.M. assisted in teratoma analysis. J.W. contributed control iPSC lines. R.I.K. provided expert input, patient samples and 31P nuclear magnetic resonance data. K.K.P. and W.T.P. supervised the study. W.T.P. wrote the manuscript, and it was revised by K.K.P., G.W. and M.L.M.

Corresponding authors

Correspondence to Kevin Kit Parker or William T Pu.

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Competing interests

J.W. is an employee of Allele Biotechnology & Pharmaceuticals.

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Wang, G., McCain, M., Yang, L. et al. Modeling the mitochondrial cardiomyopathy of Barth syndrome with induced pluripotent stem cell and heart-on-chip technologies. Nat Med 20, 616–623 (2014).

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