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:

Restriction of intestinal stem cell expansion and the regenerative response by YAP

Nature volume 493, pages 106–110 (2013)Cite this article

Subjects

Abstract

A remarkable feature of regenerative processes is their ability to halt proliferation once an organ’s structure has been restored. The Wnt signalling pathway is the major driving force for homeostatic self-renewal and regeneration in the mammalian intestine. However, the mechanisms that counterbalance Wnt-driven proliferation are poorly understood. Here we demonstrate in mice and humans that yes-associated protein 1 (YAP; also known as YAP1)—a protein known for its powerful growth-inducing and oncogenic properties1,2—has an unexpected growth-suppressive function, restricting Wnt signals during intestinal regeneration. Transgenic expression of YAP reduces Wnt target gene expression and results in the rapid loss of intestinal crypts. In addition, loss of YAP results in Wnt hypersensitivity during regeneration, leading to hyperplasia, expansion of intestinal stem cells and niche cells, and formation of ectopic crypts and microadenomas. We find that cytoplasmic YAP restricts elevated Wnt signalling independently of the AXIN–APC–GSK-3β complex partly by limiting the activity of dishevelled (DVL). DVL signals in the nucleus of intestinal stem cells, and its forced expression leads to enhanced Wnt signalling in crypts. YAP dampens Wnt signals by restricting DVL nuclear translocation during regenerative growth. Finally, we provide evidence that YAP is silenced in a subset of highly aggressive and undifferentiated human colorectal carcinomas, and that its expression can restrict the growth of colorectal carcinoma xenografts. Collectively, our work describes a novel mechanistic paradigm for how proliferative signals are counterbalanced in regenerating tissues. Additionally, our findings have important implications for the targeting of YAP in human malignancies.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: YAP overabundance inhibits Wnt-mediated intestinal regeneration.
Figure 2: Loss of YAP leads to hyperactive Wnt signalling and expansion of the stem cell niche after injury or stimulation with Rspo1.
Figure 3: YAP restricts Wnt signalling by blocking DVL nuclear translocation.
Figure 4: YAP function in human colorectal cancer.

Similar content being viewed by others

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

Microarray data have been deposited in the Gene Expression Omnibus database under accession number GSE41509.

References

  1. Pan, D. The Hippo signaling pathway in development and cancer. Dev. Cell 19, 491–505 (2010)

    Article  CAS  Google Scholar 

  2. Ramos, A. & Camargo, F. D. The Hippo signaling pathway and stem cell biology. Trends Cell Biol. 22, 339–346 (2012)

    Article  CAS  Google Scholar 

  3. Camargo, F. D. et al. YAP1 increases organ size and expands undifferentiated progenitor cells. Curr. Biol. 17, 2054–2060 (2007)

    Article  CAS  Google Scholar 

  4. Dong, J. et al. Elucidation of a universal size-control mechanism in Drosophila and mammals. Cell 130, 1120–1133 (2007)

    Article  CAS  Google Scholar 

  5. Zhang, J. et al. YAP-dependent induction of amphiregulin identifies a non-cell-autonomous component of the Hippo pathway. Nature Cell Biol. 11, 1444–1450 (2009)

    Article  CAS  Google Scholar 

  6. Roth, S. et al. Generation of a tightly regulated doxycycline-inducible model for studying mouse intestinal biology. Genesis 47, 7–13 (2009)

    Article  CAS  Google Scholar 

  7. Pinto, D., Gregorieff, A., Begthel, H. & Clevers, H. Canonical Wnt signals are essential for homeostasis of the intestinal epithelium. Genes Dev. 17, 1709–1713 (2003)

    Article  CAS  Google Scholar 

  8. Andreu, P. et al. A genetic study of the role of the Wnt/β-catenin signalling in Paneth cell differentiation. Dev. Biol. 324, 288–296 (2008)

    Article  CAS  Google Scholar 

  9. Sato, T. et al. Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. Nature 469, 415–418 (2011)

    Article  ADS  CAS  Google Scholar 

  10. van der Flier, L. G. et al. Transcription factor achaete scute-like 2 controls intestinal stem cell fate. Cell 136, 903–912 (2009)

    Article  CAS  Google Scholar 

  11. Muñoz, J. et al. The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent ‘+4’ cell markers. EMBO J. 31, 3079–3091 (2012)

    Article  Google Scholar 

  12. Fevr, T., Robine, S., Louvard, D. & Huelsken, J. Wnt/β-catenin is essential for intestinal homeostasis and maintenance of intestinal stem cells. Mol. Cell. Biol. 27, 7551–7559 (2007)

    Article  CAS  Google Scholar 

  13. Cai, J. et al. The Hippo signaling pathway restricts the oncogenic potential of an intestinal regeneration program. Genes Dev. 24, 2383–2388 (2010)

    Article  CAS  Google Scholar 

  14. Ashton, G. H. et al. Focal adhesion kinase is required for intestinal regeneration and tumorigenesis downstream of Wnt/c-Myc signaling. Dev. Cell 19, 259–269 (2010)

    Article  CAS  Google Scholar 

  15. Davies, P. S., Dismuke, A. D., Powell, A. E., Carroll, K. H. & Wong, M. H. Wnt-reporter expression pattern in the mouse intestine during homeostasis. BMC Gastroenterol. 8, 57 (2008)

    Article  Google Scholar 

  16. Ootani, A. et al. Sustained in vitro intestinal epithelial culture within a Wnt-dependent stem cell niche. Nature Med. 15, 701–706 (2009)

    Article  CAS  Google Scholar 

  17. Kim, K. A. et al. Mitogenic influence of human R-spondin1 on the intestinal epithelium. Science 309, 1256–1259 (2005)

    Article  ADS  CAS  Google Scholar 

  18. Zhao, B. et al. TEAD mediates YAP-dependent gene induction and growth control. Genes Dev. 22, 1962–1971 (2008)

    Article  CAS  Google Scholar 

  19. Schlegelmilch, K. et al. Yap1 acts downstream of α-catenin to control epidermal proliferation. Cell 144, 782–795 (2011)

    Article  CAS  Google Scholar 

  20. Sansom, O. J. et al. Loss of Apc in vivo immediately perturbs Wnt signaling, differentiation, and migration. Genes Dev. 18, 1385–1390 (2004)

    Article  CAS  Google Scholar 

  21. Imajo, M., Miyatake, K., Iimura, A., Miyamoto, A. & Nishida, E. A molecular mechanism that links Hippo signalling to the inhibition of Wnt/β-catenin signalling. EMBO J. 31, 1109–1122 (2012)

    Article  CAS  Google Scholar 

  22. Cheung, A. F. et al. Complete deletion of Apc results in severe polyposis in mice. Oncogene 29, 1857–1864 (2010)

    Article  CAS  Google Scholar 

  23. Varelas, X. et al. The Hippo pathway regulates Wnt/β-catenin signaling. Dev. Cell 18, 579–591 (2010)

    Article  CAS  Google Scholar 

  24. Itoh, K., Brott, B. K., Bae, G. U., Ratcliffe, M. J. & Sokol, S. Y. Nuclear localization is required for Dishevelled function in Wnt/β-catenin signaling. J. Biol. 4, 3 (2005)

    Article  Google Scholar 

  25. Gan, X. Q. et al. Nuclear Dvl, c-Jun, β-catenin, and TCF form a complex leading to stabilization of β-catenin–TCF interaction. J. Cell Biol. 180, 1087–1100 (2008)

    Article  CAS  Google Scholar 

  26. Metcalfe, C. et al. Dvl2 promotes intestinal length and neoplasia in the ApcMin mouse model for colorectal cancer. Cancer Res. 70, 6629–6638 (2010)

    Article  CAS  Google Scholar 

  27. Van der Flier, L. G. et al. The intestinal Wnt/TCF signature. Gastroenterology 132, 628–632 (2007)

    Article  CAS  Google Scholar 

  28. Chan, A. T., Ogino, S. & Fuchs, C. S. Aspirin and the risk of colorectal cancer in relation to the expression of COX-2. N. Engl. J. Med. 356, 2131–2142 (2007)

    Article  CAS  Google Scholar 

  29. Konsavage, W. J., Kyler, S. L., Rennoll, S. A., Jin, G. & Yochum, G. S. Wnt/β-catenin signaling regulates Yes-associated protein (YAP) gene expression in colorectal carcinoma cells. J. Biol. Chem. 287, 11730–11739 (2012)

    Article  CAS  Google Scholar 

  30. Avruch, J., Zhou, D. & Bardeesy, N. YAP oncogene overexpression supercharges colon cancer proliferation. Cell Cycle 11, 1090–1096 (2012)

    Article  CAS  Google Scholar 

  31. Madison, B. B. et al. cis elements of the villin gene control expression in restricted domains of the vertical (crypt) and horizontal (duodenum, cecum) axes of the intestine. J. Biol. Chem. 277, 33275–33283 (2002)

    Article  CAS  Google Scholar 

  32. El Marjou, F. et al. Tissue-specific and inducible Cre-mediated recombination in the gut epithelium. Genesis 39, 186–193 (2004)

    Article  CAS  Google Scholar 

  33. Barker, N. et al. Identification of stem cells in small intestine and colon by marker gene Lgr5 . Nature 449, 1003–1007 (2007)

    Article  ADS  CAS  Google Scholar 

  34. Colnot, S. et al. Colorectal cancers in a new mouse model of familial adenomatous polyposis: influence of genetic and environmental modifiers. Lab. Invest. 84, 1619–1630 (2004)

    Article  CAS  Google Scholar 

  35. Barker, N. et al. Crypt stem cells as the cells-of-origin of intestinal cancer. Nature 457, 608–611 (2009)

    Article  ADS  CAS  Google Scholar 

  36. Koo, B. K. et al. Controlled gene expression in primary Lgr5 organoid cultures. Nature Methods 9, 81–83 (2012)

    Article  CAS  Google Scholar 

  37. Sato, T. et al. Single Lgr5 stem cells build crypt–villus structures in vitro without a mesenchymal niche. Nature 459, 262–265 (2009)

    Article  ADS  CAS  Google Scholar 

  38. Meerbrey, K. L. et al. The pINDUCER lentiviral toolkit for inducible RNA interference in vitro and in vivo . Proc. Natl Acad. Sci. USA 108, 3665–3670 (2011)

    Article  ADS  CAS  Google Scholar 

  39. Ogino, S. et al. Combined analysis of COX-2 and p53 expressions reveals synergistic inverse correlations with microsatellite instability and CpG island methylator phenotype in colorectal cancer. Neoplasia 8, 458–464 (2006)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank L. Zon and Camargo laboratory members for critical review of this manuscript and X. He for Myc–DVL2 constructs. This work was supported by grants from the Stand Up to Cancer-AACR initiative (F.D.C.), National Institutes of Health R01 CA131426 and AR064036 (F.D.C.) and the Harvard Stem Cell Institute (F.D.C.). F.D.C. is a Pew Scholar in the Biomedical Sciences. E.R.B. is supported by a postdoctoral fellowship from the American Cancer Society Illinois Division (PF-12-245-01-CCG). This work has also benefited from support to K.S.Y. (CIRM, and 1K08DK096048), C.J.K. (1U01DK085527), S.O. (R01 CA151993 and R01 DK091427), S.T.M. (P01CA87969) and C.S.F. (P50CA127003).

Author information

Authors and Affiliations

  1. Stem Cell Program and Department of Hematology/Oncology, Children’s Hospital, Boston, 02115, Massachusetts, USA

    Evan R. Barry, Brian L. Butler, Kriti Shrestha, Rosemarie de la Rosa & Fernando D. Camargo

  2. Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, 02138, Massachusetts, USA

    Evan R. Barry, Brian L. Butler & Fernando D. Camargo

  3. Harvard Stem Cell Institute, Cambridge, 02138, Massachusetts, USA

    Evan R. Barry & Fernando D. Camargo

  4. Department of Medical Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, 02215, Massachusetts, USA

    Teppei Morikawa, Charles S. Fuchs & Shuji Ogino

  5. Department of Medicine, Hematology Division, Stanford University, Stanford, 94305, California, USA

    Kelley S. Yan & Calvin J. Kuo

  6. Department of Medicine, Channing Laboratory, Brigham and Women’s Hospital and Harvard Medical School, Boston, 02115, Massachusetts, USA

    Charles S. Fuchs

  7. Departments of Medicine and Biomedical Engineering, The University of North Carolina at Chapel Hill, Chapel Hill, 27599, North Carolina, USA

    Scott T. Magness

  8. Department of Gastroenterology and Hepatology, Erasmus MC, 3000 CA Rotterdam, The Netherlands,

    Ron Smits

  9. Department of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston, 02115, Massachusetts, USA

    Shuji Ogino

Authors
  1. Evan R. Barry
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Teppei Morikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Brian L. Butler
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kriti Shrestha
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rosemarie de la Rosa
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kelley S. Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Charles S. Fuchs
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Scott T. Magness
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ron Smits
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Shuji Ogino
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Calvin J. Kuo
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Fernando D. Camargo
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

E.R.B. and F.D.C. designed the study. E.R.B., B.L.B., K.S., R.d.l.R. and S.T.M. performed experiments. T.M. performed immunohistochemistry and analysis of human tumour samples. K.S.Y., C.S.F., R.S., S.O. and C.J.K. provided reagents for the study.

Corresponding author

Correspondence to Fernando D. Camargo.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-11. (PDF 11027 kb)

Supplementary Data

This file contains Supplementary Table 1 with genes identified in microarrays as being misregulated. (XLS 247 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Barry, E., Morikawa, T., Butler, B. et al. Restriction of intestinal stem cell expansion and the regenerative response by YAP. Nature 493, 106–110 (2013). https://doi.org/10.1038/nature11693

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature11693

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