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.

Induced pluripotent stem cells from a spinal muscular atrophy patient

Abstract

Spinal muscular atrophy is one of the most common inherited forms of neurological disease leading to infant mortality. Patients have selective loss of lower motor neurons resulting in muscle weakness, paralysis and often death. Although patient fibroblasts have been used extensively to study spinal muscular atrophy, motor neurons have a unique anatomy and physiology which may underlie their vulnerability to the disease process. Here we report the generation of induced pluripotent stem cells from skin fibroblast samples taken from a child with spinal muscular atrophy. These cells expanded robustly in culture, maintained the disease genotype and generated motor neurons that showed selective deficits compared to those derived from the child’s unaffected mother. This is the first study to show that human induced pluripotent stem cells can be used to model the specific pathology seen in a genetically inherited disease. As such, it represents a promising resource to study disease mechanisms, screen new drug compounds and develop new therapies.

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

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Newly generated iPS cells were fully reprogrammed.
Figure 2: iPS-SMA cells show decreased SMN transcripts.
Figure 3: iPS-WT and iPS-SMA cells can generate cells in the neural lineage.
Figure 4: iPS-WT and iPS-SMA cells increase SMN protein in response to drug treatment.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

Microarray data have been deposited in GEO under accession number GSE13828.

References

  1. Lefebvre, S. et al. Identification and characterization of a spinal muscular atrophy-determining gene. Cell 80, 155–165 (1995)

    Article  CAS  Google Scholar 

  2. Coovert, D. D. et al. The survival motor neuron protein in spinal muscular atrophy. Hum. Mol. Genet. 6, 1205–1214 (1997)

    Article  CAS  Google Scholar 

  3. Crawford, T. O. & Pardo, C. A. The neurobiology of childhood spinal muscular atrophy. Neurobiol. Dis. 3, 97–110 (1996)

    Article  CAS  Google Scholar 

  4. Munsat, T. L. & Davies, K. E. International SMA consortium meeting. (26–28 June 1992, Bonn, Germany). Neuromuscul. Disord. 2, 423–428 (1992)

    Article  CAS  Google Scholar 

  5. Lorson, C. L., Hahnen, E., Androphy, E. J. & Wirth, B. A single nucleotide in the SMN gene regulates splicing and is responsible for spinal muscular atrophy. Proc. Natl Acad. Sci. USA 96, 6307–6311 (1999)

    Article  ADS  CAS  Google Scholar 

  6. Lefebvre, S. et al. Correlation between severity and SMN protein level in spinal muscular atrophy. Nature Genet. 16, 265–269 (1997)

    Article  MathSciNet  CAS  Google Scholar 

  7. Schmid, A. & DiDonato, C. J. Animal models of spinal muscular atrophy. J. Child Neurol. 22, 1004–1012 (2007)

    Article  Google Scholar 

  8. Schrank, B. et al. Inactivation of the survival motor neuron gene, a candidate gene for human spinal muscular atrophy, leads to massive cell death in early mouse embryos. Proc. Natl Acad. Sci. USA 94, 9920–9925 (1997)

    Article  ADS  CAS  Google Scholar 

  9. DiDonato, C. J. et al. Cloning, characterization, and copy number of the murine survival motor neuron gene: homolog of the spinal muscular atrophy-determining gene. Genome Res. 7, 339–352 (1997)

    Article  CAS  Google Scholar 

  10. Hsieh-Li, H. M. et al. A mouse model for spinal muscular atrophy. Nature Genet. 24, 66–70 (2000)

    Article  CAS  Google Scholar 

  11. Monani, U. R. et al. The human centromeric survival motor neuron gene (SMN2) rescues embryonic lethality in Smn-/- mice and results in a mouse with spinal muscular atrophy. Hum. Mol. Genet. 9, 333–339 (2000)

    Article  CAS  Google Scholar 

  12. Le, T. T. et al. SMNΔ7, the major product of the centromeric survival motor neuron (SMN2) gene, extends survival in mice with spinal muscular atrophy and associates with full-length SMN. Hum. Mol. Genet. 14, 845–857 (2005)

    Article  CAS  Google Scholar 

  13. 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 

  14. Jaenisch, R. & Young, R. Stem cells, the molecular circuitry of pluripotency and nuclear reprogramming. Cell 132, 567–582 (2008)

    Article  CAS  Google Scholar 

  15. 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 

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

    Article  ADS  CAS  Google Scholar 

  17. Lowry, W. E. et al. Generation of human induced pluripotent stem cells from dermal fibroblasts. Proc. Natl Acad. Sci. USA 105, 2883–2888 (2008)

    Article  ADS  CAS  Google Scholar 

  18. Dimos, J. T. et al. Induced pluripotent stem cells generated from patients with ALS can be differentiated into motor neurons. Science 321, 1218–1221 (2008)

    Article  ADS  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  20. Jakel, R. J., Schneider, B. L. & Svendsen, C. N. Using human neural stem cells to model neurological disease. Nature Rev. Genet. 5, 136–144 (2004)

    Article  CAS  Google Scholar 

  21. Gavrilov, D. K., Shi, X. Y., Das, K., Gilliam, T. C. & Wang, C. H. Differential SMN2 expression associated with SMA severity. Nature Genet. 20, 230–231 (1998)

    Article  CAS  Google Scholar 

  22. Sumner, C. J. et al. SMN mRNA and protein levels in peripheral blood: biomarkers for SMA clinical trials. Neurology 66, 1067–1073 (2006)

    Article  CAS  Google Scholar 

  23. Fox, V. et al. Cell-cell signaling through NOTCH regulates human embryonic stem cell proliferation. Stem Cells 26, 715–723 (2008)

    Article  CAS  Google Scholar 

  24. Svendsen, C. N. et al. A new method for the rapid and long term growth of human neural precursor cells. J. Neurosci. Methods 85, 141–152 (1998)

    Article  CAS  Google Scholar 

  25. Lendahl, U., Zimmerman, L. B. & McKay, R. D. G. Cns stem-cells express a new class of intermediate filament protein. Cell 60, 585–595 (1990)

    Article  CAS  Google Scholar 

  26. Li, X. J. et al. Specification of motoneurons from human embryonic stem cells. Nature Biotechnol. 23, 215–221 (2005)

    Article  Google Scholar 

  27. Jessell, T. M. Neuronal specification in the spinal cord: Inductive signals and transcriptional codes. Nature Rev. Genet. 1, 20–29 (2000)

    Article  CAS  Google Scholar 

  28. Wichterle, H., Lieberam, I., Porter, J. A. & Jessell, T. M. Directed differentiation of embryonic stem cells into motor neurons. Cell 110, 385–397 (2002)

    Article  CAS  Google Scholar 

  29. Carriedo, S. G., Yin, H. Z. & Weiss, J. H. Motor neurons are selectively vulnerable to AMPA/kainate receptor-mediated injury in vitro . J. Neurosci. 16, 4069–4079 (1996)

    Article  CAS  Google Scholar 

  30. Monani, U. R. Spinal muscular atrophy: a deficiency in a ubiquitous protein; a motor neuron-specific disease. Neuron 48, 885–896 (2005)

    Article  CAS  Google Scholar 

  31. Brichta, L. et al. Valproic acid increases the SMN2 protein level: a well-known drug as a potential therapy for spinal muscular atrophy. Hum. Mol. Genet. 12, 2481–2489 (2003)

    Article  CAS  Google Scholar 

  32. Sumner, C. J. et al. Valproic acid increases SMN levels in spinal muscular atrophy patient cells. Ann. Neurol. 54, 647–654 (2003)

    Article  CAS  Google Scholar 

  33. Wolstencroft, E. C., Mattis, V., Bajer, A. A., Young, P. J. & Lorson, C. L. A non-sequence-specific requirement for SMN protein activity: the role of aminoglycosides in inducing elevated SMN protein levels. Hum. Mol. Genet. 14, 1199–1210 (2005)

    Article  CAS  Google Scholar 

  34. Pellizzoni, L., Yong, J. & Dreyfuss, G. Essential role for the SMN complex in the specificity of snRNP assembly. Science 298, 1775–1779 (2002)

    Article  ADS  CAS  Google Scholar 

  35. Fischer, U., Liu, Q. & Dreyfuss, G. The SMN–SIP1 complex has an essential role in spliceosomal snRNP biogenesis. Cell 90, 1023–1029 (1997)

    Article  CAS  Google Scholar 

  36. Liu, Q., Fischer, U., Wang, F. & Dreyfuss, G. The spinal muscular atrophy disease gene product, SMN, and its associated protein SIP1 are in a complex with spliceosomal snRNP proteins. Cell 90, 1013–1021 (1997)

    Article  CAS  Google Scholar 

  37. Carrel, T. L. et al. Survival motor neuron function in motor axons is independent of functions required for small nuclear ribonucleoprotein biogenesis. J. Neurosci. 26, 11014–11022 (2006)

    Article  CAS  Google Scholar 

  38. Zhang, H. et al. Multiprotein complexes of the survival of motor neuron protein SMN with gemins traffic to neuronal processes and growth cones of motor neurons. J. Neurosci. 26, 8622–8632 (2006)

    Article  CAS  Google Scholar 

  39. Bolstad, B. M., Irizarry, R. A., Astrand, M. & Speed, T. P. A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 19, 185–193 (2003)

    Article  CAS  Google Scholar 

  40. Irizarry, R. A. et al. Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4, 249–264 (2003)

    Article  Google Scholar 

  41. Suzuki, R. & Shimodaira, H. Pvclust: an R package for assessing the uncertainty in hierarchical clustering. Bioinformatics 22, 1540–1542 (2006)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank J. Meyer for helpful discussions and B. Shelley, B. Heins and E. McMillan for technical assistance. We also thank WiCell Research Institute for karyotype analysis, Cell Line Genetics for DNA fingerprinting, R. Stewart, S. Tian and V. Ruotti at the Morgridge Institute for Research for microarray analysis, and Promega Corp. for qRT–PCR analysis (all at Madison, Wisconsin). The MNR2/HB9 (81.5C10) and the ISLET1 (40.2D6) monoclonal antibodies (both developed by T. Jessell) were obtained from the Developmental Studies Hybridoma Bank. Funding support was provided by the Amyotrophic Lateral Sclerosis Association (to C.N.S.), the National Institutes of Neurological Disorders and Stroke (P01NS057778 to C.N.S. and R01NS41584 to C.L.L.), National Institutes of Child Health and Human Development (R01HD054413 to C.L.L.), and National Institutes of General Medical Sciences (T32GM008396 for F.F.R.).

Author Contributions A.D.E. participated in all aspects and prepared the manuscript; J.Y. generated and aided in the characterization of iPS-SMA and iPS-WT clones; F.F.R., V.B.M. and C.L.L. performed SMN analysis and manuscript preparation; J.A.T. participated in the generation of the iPS clones; C.N.S. conceived the project and participated in planning, data analysis and manuscript preparation.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Allison D. Ebert or Clive N. Svendsen.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-7 with Legends and Supplementary Tables 1-3 (PDF 2790 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ebert, A., Yu, J., Rose, F. et al. Induced pluripotent stem cells from a spinal muscular atrophy patient. Nature 457, 277–280 (2009). https://doi.org/10.1038/nature07677

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature07677

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