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Abstract

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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Acknowledgements

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.

Author information

Author notes

    • Gang Wang
    •  & Megan L McCain

    These authors contributed equally to this work.

Affiliations

  1. Department of Cardiology, Boston Children's Hospital, Boston, Massachusetts, USA.

    • Gang Wang
    • , Aibin He
    • , Dawei Jiang
    • , Donghui Zhang
    • , Lior Zangi
    • , Judith Geva
    • , Amy E Roberts
    • , Qing Ma
    • , Jian Ding
    • , Jinghai Chen
    • , Da-Zhi Wang
    • , Kai Li
    •  & William T Pu
  2. Wyss Institute for Biologically Inspired Engineering, School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA.

    • Megan L McCain
    • , Luhan Yang
    • , Francesco Silvio Pasqualini
    • , Ashutosh Agarwal
    • , Hongyan Yuan
    • , George M Church
    •  & Kevin Kit Parker
  3. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.

    • Luhan Yang
    •  & George M Church
  4. Department of Medicine, Division of Genetics, Boston Children's Hospital, Boston, Massachusetts, USA.

    • Amy E Roberts
  5. Allele Biotechnology & Pharmaceuticals, Inc., San Diego, California, USA.

    • Jiwu Wang
  6. Department of Photobiology and Bioengineering, The Scintillon Institute, San Diego, California, USA.

    • Jiwu Wang
  7. Department of Clinical Chemistry and Pediatrics, Laboratory of Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands.

    • Ronald J A Wanders
    • , Wim Kulik
    •  & Frédéric M Vaz
  8. Department of Pathology, Center for Cardiovascular Biology and Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, Washington, USA.

    • Michael A Laflamme
    •  & Charles E Murry
  9. Department of Bioengineering, Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, Washington, USA.

    • Charles E Murry
  10. Department of Medicine and Cardiology, Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, Washington, USA.

    • Charles E Murry
  11. Department of Cell and Molecular Biology and Medicine, Karolinska Institutet, Stockholm, Sweden.

    • Kenneth R Chien
  12. Division of Metabolism, Kennedy Krieger Institute, Baltimore, Maryland, USA.

    • Richard I Kelley
  13. Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, USA.

    • Kevin Kit Parker
    •  & William T Pu

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Contributions

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.

Competing interests

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

Corresponding authors

Correspondence to Kevin Kit Parker or William T Pu.

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DOI

https://doi.org/10.1038/nm.3545