Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Modelling pathogenesis and treatment of familial dysautonomia using patient-specific iPSCs

Abstract

The isolation of human induced pluripotent stem cells (iPSCs)1,2,3 offers a new strategy for modelling human disease. Recent studies have reported the derivation and differentiation of disease-specific human iPSCs4,5,6,7. However, a key challenge in the field is the demonstration of disease-related phenotypes and the ability to model pathogenesis and treatment of disease in iPSCs. Familial dysautonomia (FD) is a rare but fatal peripheral neuropathy, caused by a point mutation in the IKBKAP8 gene involved in transcriptional elongation9. The disease is characterized by the depletion of autonomic and sensory neurons. The specificity to the peripheral nervous system and the mechanism of neuron loss in FD are poorly understood owing to the lack of an appropriate model system. Here we report the derivation of patient-specific FD-iPSCs and the directed differentiation into cells of all three germ layers including peripheral neurons. Gene expression analysis in purified FD-iPSC-derived lineages demonstrates tissue-specific mis-splicing of IKBKAP in vitro. Patient-specific neural crest precursors express particularly low levels of normal IKBKAP transcript, suggesting a mechanism for disease specificity. FD pathogenesis is further characterized by transcriptome analysis and cell-based assays revealing marked defects in neurogenic differentiation and migration behaviour. Furthermore, we use FD-iPSCs for validating the potency of candidate drugs in reversing aberrant splicing and ameliorating neuronal differentiation and migration. Our study illustrates the promise of iPSC technology for gaining new insights into human disease pathogenesis and treatment.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Establishment of FD-iPSCs from patient fibroblasts.
Figure 2: FD-iPSC-derived cell lineages model the tissue specificity of FD IKBKAP splicing defect.
Figure 3: Molecular and functional characterization of FD-iPSC-derived neural crest precursor cells.
Figure 4: Validating kinetin as a candidate compound for treating FD-iPSC-derived neural crest cells.

Similar content being viewed by others

References

  1. Yu, J. et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 318, 1917–1920 (2007)

    Article  ADS  CAS  Google Scholar 

  2. Takahashi, K. et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872 (2007)

    Article  CAS  Google Scholar 

  3. Park, I. H. et al. Reprogramming of human somatic cells to pluripotency with defined factors. Nature 451, 141–146 (2008)

    Article  ADS  CAS  Google Scholar 

  4. Dimos, J. T. et al. Induced pluripotent stem cells generated from patients with ALS can be differentiated into motor neurons. Science 131, 1218–1221 (2008)

    Article  ADS  Google Scholar 

  5. Ebert, A. D. et al. Induced pluripotent stem cells from a spinal muscular atrophy patient. Nature 457, 277–280 (2009)

    Article  ADS  CAS  Google Scholar 

  6. Park, I. H. et al. Disease-specific induced pluripotent stem cells. Cell 134, 877–886 (2008)

    Article  CAS  Google Scholar 

  7. Soldner, F. et al. Parkinson’s disease patient-derived induced pluripotent stem cells free of viral reprogramming factors. Cell 136, 964–977 (2009)

    Article  CAS  Google Scholar 

  8. Slaugenhaupt, S. A. et al. Tissue-specific expression of a splicing mutation in the IKBKAP gene causes familial dysautonomia. Am. J. Hum. Genet. 68, 598–605 (2001)

    Article  CAS  Google Scholar 

  9. Close, P. et al. Transcription impairment and cell migration defects in elongator-depleted cells: implication for familial dysautonomia. Mol. Cell 22, 521–531 (2006)

    Article  CAS  Google Scholar 

  10. Axelrod, F. B., Goldberg, J. D., Ye, X. Y. & Maayan, C. Survival in familial dysautonomia: impact of early intervention. J. Pediatr. 141, 518–523 (2002)

    Article  Google Scholar 

  11. Anderson, S. L. et al. Familial dysautonomia is caused by mutations of the IKAP gene. Am. J. Hum. Genet. 68, 753–758 (2001)

    Article  CAS  Google Scholar 

  12. Slaugenhaupt, S. A. et al. Rescue of a human mRNA splicing defect by the plant cytokinin kinetin. Hum. Mol. Genet. 13, 429–436 (2004)

    Article  CAS  Google Scholar 

  13. Papapetrou, E. P. et al. Stoichiometric and temporal requirements of Oct4, Sox2, Klf4 and cMyc expression for efficient human iPSC induction and differentiation. Proc. Natl Acad. Sci. USA 106, 12759–12764 (2009)

    Article  ADS  CAS  Google Scholar 

  14. Elkabetz, Y. et al. Human ES cell-derived neural rosettes reveal a functionally dinstinct early neural stem cell stage. Genes Dev. 22, 152–165 (2008)

    Article  CAS  Google Scholar 

  15. Lee, G. et al. Isolation and directed differentiation of neural crest stem cells derived from human embryonic stem cells. Nature Biotechnol. 25, 1468–1475 (2007)

    Article  CAS  Google Scholar 

  16. Lee, G. S., Kim, B. S., Sheih, J. H. & Moore, M. Forced expression of HoxB4 enhances hematopoietic differentiation by human embryonic stem cells. Mol. Cells 25, 487–493 (2008)

    CAS  PubMed  Google Scholar 

  17. Lu, S. J. et al. Generation of functional hemangioblasts from human embryonic stem cells. Nature Methods 4, 501–509 (2007)

    Article  CAS  Google Scholar 

  18. D’Amour, K. A. et al. Efficient differentiation of human embryonic stem cells to definitive endoderm. Nature Biotechnol. 23, 1534–1541 (2005)

    Article  Google Scholar 

  19. Iwashita, T., Kruger, G. M., Pardal, R., Kiel, M. J. & Morrison, S. J. Hirschsprung disease is linked to defects in neural crest stem cell function. Science 301, 972–976 (2003)

    Article  ADS  CAS  Google Scholar 

  20. Jones, N. C. et al. Prevention of the neurocristopathy Treacher Collins syndrome through inhibition of p53 function. Nature Med. 14, 125–133 (2008)

    Article  CAS  Google Scholar 

  21. Sommer, L., Shah, N., Rao, M. & Anderson, D. J. The cellular function of MASH1 in autonomic neurogenesis. Neuron 15, 1245–1258 (1995)

    Article  CAS  Google Scholar 

  22. Johansen, L. D. et al. IKAP localizes to membrane ruffles with filamin A and regulates actin cytoskeleton organization and cell migration. J. Cell Sci. 121, 854–864 (2008)

    Article  CAS  Google Scholar 

  23. Anderson, S. L., Qiu, J. & Rubin, B. Y. EGCG corrects aberrant splicing of IKAP mRNA in cells from patients with familial dysautonomia. Biochem. Biophys. Res. Commun. 310, 627–633 (2003)

    Article  CAS  Google Scholar 

  24. Anderson, S. L., Qiu, J. & Rubin, B. Y. Tocotrienols induce IKBKAP expression: a possible therapy for familial dysautonomia. Biochem. Biophys. Res. Commun. 306, 303–309 (2003)

    Article  CAS  Google Scholar 

  25. Axelrod, F. B. Familial dysautonomia: a review of the current pharmacological treatments. Expert Opin. Pharmacother. 6, 561–567 (2005)

    Article  CAS  Google Scholar 

  26. Perrier, A. L. et al. From the Cover: derivation of midbrain dopamine neurons from human embryonic stem cells. Proc. Natl Acad. Sci. USA 101, 12543–12548 (2004)

    Article  ADS  CAS  Google Scholar 

  27. Placantonakis, D. G. et al. BAC transgenesis in human ES cells as a novel tool to define the human neural lineage. Stem Cells 27, 521–532 (2009)

    Article  CAS  Google Scholar 

  28. Chambers, S. M. et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nature Biotechnol. 27, 275–280 (2009)

    Article  CAS  Google Scholar 

  29. Kennedy, M., D’Souza, S. L., Lynch-Kattman, M., Schwantz, S. & Keller, G. Development of the hemangioblast defines the onset of hematopoiesis in human ES cell differentiation cultures. Blood 109, 2679–2687 (2007)

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Rodriguez, L. G., Wu, X. & Guan, J. L. Wound-healing assay. Methods Mol. Biol. 294, 23–29 (2005)

    PubMed  Google Scholar 

Download references

Acknowledgements

We thank J. Hendrikx, M. Leversha and C. Zhao for technical help. The work was supported by grants from the Starr Foundation and NYSTEM, by the New York Stem Cell Foundation (NYCSF, Druckenmiller fellowships to G.L. and C.A.F.) and by the Starr Scholar fellowship to S.M.C.

Author Contributions G.L.: conception and study design, maintenance and directed differentiation of iPSCs, cellular/molecular assays for disease modelling, data assembly, analysis and interpretation, and writing of manuscript; E.P.P., H.K. and C.A.F.: iPSC clone derivation and maintenance; S.M.C., M.J.T. and A.V.: data collection, analysis and interpretation; Y.M.G., J.M. and F.S.: in vivo experiments and histological analyses; V.T. and M.S.: study design, data analysis and interpretation; L.S.: conception and study design, data analysis and interpretation, and writing of manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lorenz Studer.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-11 with Legends and Supplementary Tables 1-3. (PDF 3340 kb)

Supplementary Movie 1

This movie, shows in real-time beating putative cardiomyocytes derived from FD human iPS cell line (clone#22). (MOV 1730 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lee, G., Papapetrou, E., Kim, H. et al. Modelling pathogenesis and treatment of familial dysautonomia using patient-specific iPSCs. Nature 461, 402–406 (2009). https://doi.org/10.1038/nature08320

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature08320

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing