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.

Telomere shortening and loss of self-renewal in dyskeratosis congenita induced pluripotent stem cells

Nature volume 474, pages 399–402 (2011)Cite this article

Subjects

Abstract

The differentiation of patient-derived induced pluripotent stem cells (iPSCs) to committed fates such as neurons, muscle and liver is a powerful approach for understanding key parameters of human development and disease1,2,3,4,5,6. Whether undifferentiated iPSCs themselves can be used to probe disease mechanisms is uncertain. Dyskeratosis congenita is characterized by defective maintenance of blood, pulmonary tissue and epidermal tissues and is caused by mutations in genes controlling telomere homeostasis7,8. Short telomeres, a hallmark of dyskeratosis congenita, impair tissue stem cell function in mouse models, indicating that a tissue stem cell defect may underlie the pathophysiology of dyskeratosis congenita9,10. Here we show that even in the undifferentiated state, iPSCs from dyskeratosis congenita patients harbour the precise biochemical defects characteristic of each form of the disease and that the magnitude of the telomere maintenance defect in iPSCs correlates with clinical severity. In iPSCs from patients with heterozygous mutations in TERT, the telomerase reverse transcriptase, a 50% reduction in telomerase levels blunts the natural telomere elongation that accompanies reprogramming. In contrast, mutation of dyskerin (DKC1) in X-linked dyskeratosis congenita severely impairs telomerase activity by blocking telomerase assembly and disrupts telomere elongation during reprogramming. In iPSCs from a form of dyskeratosis congenita caused by mutations in TCAB1 (also known as WRAP53), telomerase catalytic activity is unperturbed, yet the ability of telomerase to lengthen telomeres is abrogated, because telomerase mislocalizes from Cajal bodies to nucleoli within the iPSCs. Extended culture of DKC1-mutant iPSCs leads to progressive telomere shortening and eventual loss of self-renewal, indicating that a similar process occurs in tissue stem cells in dyskeratosis congenita patients. These findings in iPSCs from dyskeratosis congenita patients reveal that undifferentiated iPSCs accurately recapitulate features of a human stem cell disease and may serve as a cell-culture-based system for the development of targeted therapeutics.

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

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Learn more

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

Figure 1: Dyskeratosis congenita iPSCs with heterozygous TERT mutations show reduced telomerase levels.
Figure 2: Preserved activity, but pronounced mislocalization of telomerase in TCAB1 -mutant iPSCs.
Figure 3: Diminished TERC levels, reduced activity and impaired assembly of mature telomerase in X-linked dyskeratosis congenita iPSCs.
Figure 4: Impaired telomere maintenance and loss of self-renewal in dyskeratosis congenita iPSCs.

References

  1. 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 

  2. Rashid, S. T. et al. Modeling inherited metabolic disorders of the liver using human induced pluripotent stem cells. J. Clin. Invest. 120, 3127–3136 (2010)

    Article  CAS  Google Scholar 

  3. Liu, G. H. et al. Recapitulation of premature ageing with iPSCs from Hutchinson–Gilford progeria syndrome. Nature advance online publication. 10.1038/nature09879 (23 February 2011)

  4. 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 

  5. 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 

  6. Maehr, R. et al. Generation of pluripotent stem cells from patients with type 1 diabetes. Proc. Natl Acad. Sci. USA 106, 15768–15773 (2009)

    Article  ADS  CAS  Google Scholar 

  7. Walne, A. J. & Dokal, I. Advances in the understanding of dyskeratosis congenita. Br. J. Haematol. 145, 164–172 (2009)

    Article  CAS  Google Scholar 

  8. Alter, B. P. et al. Malignancies and survival patterns in the National Cancer Institute inherited bone marrow failure syndromes cohort study. Br. J. Haematol. 150, 179–188 (2010)

    PubMed  PubMed Central  Google Scholar 

  9. Lee, H. W. et al. Essential role of mouse telomerase in highly proliferative organs. Nature 392, 569–574 (1998)

    Article  ADS  CAS  Google Scholar 

  10. Allsopp, R. C., Morin, G. B., DePinho, R., Harley, C. B. & Weissman, I. L. Telomerase is required to slow telomere shortening and extend replicative lifespan of HSCs during serial transplantation. Blood 102, 517–520 (2003)

    Article  CAS  Google Scholar 

  11. Bessler, M., Wilson, D. B. & Mason, P. J. Dyskeratosis congenita. FEBS Lett. 584, 3831–3838 (2010)

    Article  CAS  Google Scholar 

  12. Mitchell, J. R., Wood, E. & Collins, K. A telomerase component is defective in the human disease dyskeratosis congenita. Nature 402, 551–555 (1999)

    Article  ADS  CAS  Google Scholar 

  13. Zhong, F. et al. Disruption of telomerase trafficking by TCAB1 mutation causes dyskeratosis congenita. Genes Dev. 25, 11–16 (2011)

    Article  CAS  Google Scholar 

  14. Alter, B. P. et al. Very short telomere length by flow fluorescence in situ hybridization identifies patients with dyskeratosis congenita. Blood 110, 1439–1447 (2007)

    Article  CAS  Google Scholar 

  15. Agarwal, S. et al. Telomere elongation in induced pluripotent stem cells from dyskeratosis congenita patients. Nature 464, 292–296 (2010)

    Article  ADS  CAS  Google Scholar 

  16. Marion, R. M. et al. Telomeres acquire embryonic stem cell characteristics in induced pluripotent stem cells. Cell Stem Cell 4, 141–154 (2009)

    Article  CAS  Google Scholar 

  17. Stadtfeld, M., Maherali, N., Breault, D. T. & Hochedlinger, K. Defining molecular cornerstones during fibroblast to iPS cell reprogramming in mouse. Cell Stem Cell 2, 230–240 (2008)

    Article  CAS  Google Scholar 

  18. Yoshida, Y., Takahashi, K., Okita, K., Ichisaka, T. & Yamanaka, S. Hypoxia enhances the generation of induced pluripotent stem cells. Cell Stem Cell 5, 237–241 (2009)

    Article  CAS  Google Scholar 

  19. Wong, J. M. & Collins, K. Telomerase RNA level limits telomere maintenance in X-linked dyskeratosis congenita. Genes Dev. 20, 2848–2858 (2006)

    Article  CAS  Google Scholar 

  20. Zaug, A. J., Podell, E. R., Nandakumar, J. & Cech, T. R. Functional interaction between telomere protein TPP1 and telomerase. Genes Dev. 24, 613–622 (2010)

    Article  CAS  Google Scholar 

  21. Vulliamy, T. et al. Disease anticipation is associated with progressive telomere shortening in families with dyskeratosis congenita due to mutations in TERC. Nature Genet. 36, 447–449 (2004)

    Article  CAS  Google Scholar 

  22. Armanios, M. et al. Haploinsufficiency of telomerase reverse transcriptase leads to anticipation in autosomal dominant dyskeratosis congenita. Proc. Natl Acad. Sci. USA 102, 15960–15964 (2005)

    Article  ADS  CAS  Google Scholar 

  23. Venteicher, A. S. et al. A human telomerase holoenzyme protein required for Cajal body localization and telomere synthesis. Science 323, 644–648 (2009)

    Article  ADS  CAS  Google Scholar 

  24. Tycowski, K. T., Shu, M. D., Kukoyi, A. & Steitz, J. A. A conserved WD40 protein binds the Cajal body localization signal of scaRNP particles. Mol. Cell 34, 47–57 (2009)

    Article  CAS  Google Scholar 

  25. Lundblad, V. & Szostak, J. W. A mutant with a defect in telomere elongation leads to senescence in yeast. Cell 57, 633–643 (1989)

    Article  CAS  Google Scholar 

  26. Matera, A. G., Terns, R. M. & Terns, M. P. Non-coding RNAs: lessons from the small nuclear and small nucleolar RNAs. Nature Rev. Mol. Cell Biol. 8, 209–220 (2007)

    Article  CAS  Google Scholar 

  27. Byrne, J. A., Nguyen, H. N. & Reijo Pera, R. A. Enhanced generation of induced pluripotent stem cells from a subpopulation of human fibroblasts. PLoS ONE 4, e7118 (2009)

    Article  ADS  Google Scholar 

  28. Sommer, C. A. et al. Induced pluripotent stem cell generation using a single lentiviral stem cell cassette. Stem Cells 27, 543–549 (2009)

    Article  CAS  Google Scholar 

  29. Tomlinson, R. L. et al. Telomerase reverse transcriptase is required for the localization of telomerase RNA to Cajal bodies and telomeres in human cancer cells. Mol. Biol. Cell 19, 3793–3800 (2008)

    Article  CAS  Google Scholar 

  30. Middleman, E. J., Choi, J., Venteicher, A. S., Cheung, P. & Artandi, S. E. Regulation of cellular immortalization and steady-state levels of the telomerase reverse transcriptase through its carboxy-terminal domain. Mol. Cell. Biol. 26, 2146–2159 (2006)

    Article  CAS  Google Scholar 

  31. Cristofari, G. & Lingner, J. Telomere length homeostasis requires that telomerase levels are limiting. EMBO J. 25, 565–574 (2006)

    Article  CAS  Google Scholar 

  32. Lei, M., Zaug, A. J., Podell, E. R. & Cech, T. R. Switching human telomerase on and off with hPOT1 protein in vitro. J. Biol. Chem. 280, 20449–20456 (2005)

    Article  CAS  Google Scholar 

  33. Wang, F. et al. The POT1–TPP1 telomere complex is a telomerase processivity factor. Nature 445, 506–510 (2007)

    Article  ADS  CAS  Google Scholar 

  34. Venteicher, A. S., Meng, Z., Mason, P. J., Veenstra, T. D. & Artandi, S. E. Identification of ATPases pontin and reptin as telomerase components essential for holoenzyme assembly. Cell 132, 945–957 (2008)

    Article  CAS  Google Scholar 

  35. Mochizuki, Y., He, J., Kulkarni, S., Bessler, M. & Mason, P. J. Mouse dyskerin mutations affect accumulation of telomerase RNA and small nucleolar RNA, telomerase activity, and ribosomal RNA processing. Proc. Natl Acad. Sci. USA 101, 10756–10761 (2004)

    Article  ADS  CAS  Google Scholar 

  36. Seabright, M. Rapid banding technique for human chromosomes. Lancet 2, 971–972 (1971)

    Article  CAS  Google Scholar 

  37. Deb-Rinker, P., Ly, D., Jezierski, A., Sikorska, M. & Walker, P. R. Sequential DNA methylation of the Nanog and Oct-4 upstream regions in human NT2 cells during neuronal differentiation. J. Biol. Chem. 280, 6257–6260 (2005)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

L.F.Z.B. is the recipient of a Pew Fellowship. M.F.P. and K.T.X. are the recipients of NSF Graduate Research Fellowships. F.L.Z. was supported by A*STAR, Singapore. We thank the patients for their valuable contributions and L. Leathwood for study support. This work was supported, in part, by the intramural research program of the Division of Cancer Epidemiology and Genetics, NCI, NIH; by a CIRM Shared Research Laboratory grant to R.R.P.; and by grants from the NCI, NIA, NHLBI and CIRM to S.E.A.

Author information

Authors and Affiliations

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

    Luis F. Z. Batista, Matthew F. Pech, Franklin L. Zhong, Jinkuk Choi & Steven E. Artandi

  2. Cancer Biology Program, Stanford University School of Medicine, Stanford, 94305, California, USA

    Matthew F. Pech, Franklin L. Zhong, Jinkuk Choi, Renee A. Reijo Pera & Steven E. Artandi

  3. Department of Obstetrics and Gynecology, Institute for Stem Cell Biology & Regenerative Medicine, Stanford University School of Medicine, Stanford, 94305, California, USA

    Ha Nam Nguyen, Vittorio Sebastiano, Marius Wernig & Renee A. Reijo Pera

  4. Department of Biochemistry, Stanford University School of Medicine, Stanford, 94305, California, USA

    Kathleen T. Xie

  5. Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, University of Colorado, Boulder, 80309, Colorado, USA

    Arthur J. Zaug, Sharon M. Crary & Thomas R. Cech

  6. Department of Pathology, Stanford University School of Medicine, Stanford, 94305, California, USA

    Vittorio Sebastiano, Athena Cherry & Marius Wernig

  7. Division of Cancer Epidemiology & Genetics, Department of Health & Human Services, Clinical Genetics Branch, National Cancer Institute, National Institutes of Health, Bethesda, 20852, Maryland, USA

    Neelam Giri, Blanche P. Alter & Sharon A. Savage

  8. The Glenn Laboratories for the Biology of Aging, Stanford University School of Medicine, Stanford, 94305, California, USA

    Steven E. Artandi

Authors
  1. Luis F. Z. Batista
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Matthew F. Pech
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Franklin L. Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ha Nam Nguyen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kathleen T. Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Arthur J. Zaug
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Sharon M. Crary
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jinkuk Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Vittorio Sebastiano
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Athena Cherry
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Neelam Giri
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Marius Wernig
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Blanche P. Alter
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Thomas R. Cech
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Sharon A. Savage
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Renee A. Reijo Pera
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Steven E. Artandi
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

L.F.Z.B., M.F.P., F.L.Z and S.E.A. designed the experiments and analysed the data; L.F.Z.B., M.F.P., F.L.Z., H.N.N., K.T.X., A.J.Z., S.M.C., J.C., V.S. and A.C. performed the experiments; N.G., M.W., B.P.A., T.R.C., S.A.S. and R.A.R.P. analysed the data; N.G., B.P.A and S.A.S. collected patients’ material; L.F.Z.B. and S.E.A. wrote the manuscript.

Corresponding author

Correspondence to Steven E. Artandi.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-15 with legends, Supplementary Tables 1-2 and additional references. (PDF 1585 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Batista, L., Pech, M., Zhong, F. et al. Telomere shortening and loss of self-renewal in dyskeratosis congenita induced pluripotent stem cells. Nature 474, 399–402 (2011). https://doi.org/10.1038/nature10084

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature10084

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

Advanced 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