Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome (original) (raw)

Nature volume 471, pages 230–234 (2011)Cite this article

Abstract

Individuals with congenital or acquired prolongation of the QT interval, or long QT syndrome (LQTS), are at risk of life-threatening ventricular arrhythmia1,2. LQTS is commonly genetic in origin but can also be caused or exacerbated by environmental factors1,3. A missense mutation in the L-type calcium channel CaV1.2 leads to LQTS in patients with Timothy syndrome4,5. To explore the effect of the Timothy syndrome mutation on the electrical activity and contraction of human cardiomyocytes, we reprogrammed human skin cells from Timothy syndrome patients to generate induced pluripotent stem cells, and differentiated these cells into cardiomyocytes. Electrophysiological recording and calcium (Ca2+) imaging studies of these cells revealed irregular contraction, excess Ca2+ influx, prolonged action potentials, irregular electrical activity and abnormal calcium transients in ventricular-like cells. We found that roscovitine, a compound that increases the voltage-dependent inactivation of CaV1.2 (refs 6–8), restored the electrical and Ca2+ signalling properties of cardiomyocytes from Timothy syndrome patients. This study provides new opportunities for studying the molecular and cellular mechanisms of cardiac arrhythmias in humans, and provides a robust assay for developing new drugs to treat these diseases.

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Acknowledgements

We thank K. Timothy and the Timothy syndrome patients who participated in this study; U. Francke for discussion and for providing karyotyping expertise; A. Cherry and D. Bangs for fibroblast isolation; K. C. Chan for iPSC cultures; O. Shcheglovitov for help with electrophysiological recordings; and A. Olson for help with the confocal microscope. Funding was provided by grants from the Japan Society for the Promotion for Science and the American Heart Association Western States to M.Y., and a National Institutes of Health Director’s Pioneer Award, a grant from the Simons Foundation to R.E.D and gifts from L. Miller, B. and F. Horowitz and M. McCaffery.

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Author notes

  1. Brian Hsueh, Xiaolin Jia & Anca M. Pasca
    Present address: Present addresses: Department of Chemistry, Princeton University, Princeton, New Jersey 08544, USA (B.H.); Baylor College of Medicine, Houston, Texas 77030, USA (X.J.); Lucile Packard Children's Hospital, Stanford University, Palo Alto, California 94304, USA (A.M.P.).,

Authors and Affiliations

  1. Department of Neurobiology, Stanford University School of Medicine, Stanford, 94305, California, USA
    Masayuki Yazawa, Brian Hsueh, Xiaolin Jia, Anca M. Pasca & Ricardo E. Dolmetsch
  2. Department of Pediatrics, Stanford University School of Medicine, Stanford, 94305, California, USA
    Jonathan A. Bernstein
  3. Department of Psychiatry & Behavioral Science, Stanford University School of Medicine, Stanford, 94305, California, USA
    Joachim Hallmayer

Authors

  1. Masayuki Yazawa
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  2. Brian Hsueh
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  3. Xiaolin Jia
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  4. Anca M. Pasca
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  5. Jonathan A. Bernstein
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  6. Joachim Hallmayer
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  7. Ricardo E. Dolmetsch
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Contributions

M.Y. and R.E.D. designed research and wrote the manuscript; J.A.B., J.H. and R.E.D recruited the Timothy syndrome patients; M.Y. and X.J. generated and characterized control and Timothy syndrome iPSCs; A.M.P. conducted karyotyping; M.Y. performed generation and characterization of human cardiomyocytes, whole-cell patch clamp, and Ca2+ imaging; M.Y. and B.H. analysed cardiomyocytes contraction rates.

Corresponding author

Correspondence toRicardo E. Dolmetsch.

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The authors declare no competing financial interests.

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Yazawa, M., Hsueh, B., Jia, X. et al. Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome.Nature 471, 230–234 (2011). https://doi.org/10.1038/nature09855

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Editorial Summary

New model for arrhythmias

It is difficult to model cardiac arrhythmias in mice and other genetically tractable animals because the mechanisms of cardiomyocyte contraction in these animals are unlike those in humans. A new model for studying these conditions is reported, in the form of cardiomyocytes produced from induced pluripotent stem cells derived by reprogramming fibroblasts from two patients with Timothy syndrome, a disorder characterized by autism, immune deficiency and cardiac arrhythmias. The abnormal electrical and calcium-signalling properties of these patients' cells were restored by a drug, roscovitine, known to increase voltage-dependent inactivation of CaV1.2, a calcium channel that is defective in patients with Timothy syndrome.