Letter | Published:

Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome

Nature 471, 230–234 (10 March 2011) | Download Citation

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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.

Author information

    • Brian Hsueh
    • , Xiaolin Jia
    •  & Anca M. Pasca

    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.).

Affiliations

  1. Department of Neurobiology, Stanford University School of Medicine, Stanford, California 94305, USA

    • Masayuki Yazawa
    • , Brian Hsueh
    • , Xiaolin Jia
    • , Anca M. Pasca
    •  & Ricardo E. Dolmetsch

  2. Department of Pediatrics, Stanford University School of Medicine, Stanford, California 94305, USA

    • Jonathan A. Bernstein

  3. Department of Psychiatry & Behavioral Science, Stanford University School of Medicine, Stanford, California 94305, USA

    • Joachim Hallmayer

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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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Ricardo E. Dolmetsch.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-8 with legends and Supplementary Tables 1-2.

  2. 2.

    Supplementary Methods

    This file contains Supplementary Materials and Methods.

Videos

  1. 1.

    Supplementary Movie 1

    This movie shows control EBs (round) spontaneously contracting at d37.

  2. 2.

    Supplementary Movie 2

    This movie shows control EBs (flat) spontaneously contracting at d37.

  3. 3.

    Supplementary Movie 3

    This movie shows TS EBs (round) spontaneously contracting at d37.

  4. 4.

    Supplementary Movie 4

    This movie shows TS EBs (flat) spontaneously contracting at d37.

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DOI

https://doi.org/10.1038/nature09855

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