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:

FADD prevents RIP3-mediated epithelial cell necrosis and chronic intestinal inflammation

Abstract

Intestinal immune homeostasis depends on a tightly regulated cross talk between commensal bacteria, mucosal immune cells and intestinal epithelial cells (IECs)1,2,3,4. Epithelial barrier disruption is considered to be a potential cause of inflammatory bowel disease; however, the mechanisms regulating intestinal epithelial integrity are poorly understood1,5. Here we show that mice with IEC-specific knockout of FADD (FADDIEC-KO), an adaptor protein required for death-receptor-induced apoptosis6, spontaneously developed epithelial cell necrosis, loss of Paneth cells, enteritis and severe erosive colitis. Genetic deficiency in RIP3, a critical regulator of programmed necrosis7,8,9, prevented the development of spontaneous pathology in both the small intestine and colon of FADDIEC-KO mice, demonstrating that intestinal inflammation is triggered by RIP3-dependent death of FADD-deficient IECs. Epithelial-specific inhibition of CYLD, a deubiquitinase that regulates cellular necrosis10, prevented colitis development in FADDIEC-KO but not in NEMOIEC-KO mice11, showing that different mechanisms mediated death of colonic epithelial cells in these two models. In FADDIEC-KO mice, TNF deficiency ameliorated colon inflammation, whereas MYD88 deficiency and also elimination of the microbiota prevented colon inflammation, indicating that bacteria-mediated Toll-like-receptor signalling drives colitis by inducing the expression of TNF and other cytokines. However, neither CYLD, TNF or MYD88 deficiency nor elimination of the microbiota could prevent Paneth cell loss and enteritis in FADDIEC-KO mice, showing that different mechanisms drive RIP3-dependent necrosis of FADD-deficient IECs in the small and large bowel. Therefore, by inhibiting RIP3-mediated IEC necrosis, FADD preserves epithelial barrier integrity and antibacterial defence, maintains homeostasis and prevents chronic intestinal inflammation. Collectively, these results show that mechanisms preventing RIP3-mediated epithelial cell death are critical for the maintenance of intestinal homeostasis and indicate that programmed necrosis of IECs might be implicated in the pathogenesis of inflammatory bowel disease, in which Paneth cell and barrier defects are thought to contribute to intestinal inflammation.

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: Mice with IEC-specific ablation of FADD spontaneously develop severe colitis.
Figure 2: RIP3- and CYLD-dependent necrosis of IECs triggers colitis in FADD IEC-KO mice.
Figure 3: Spontaneous colitis development in FADD IEC-KO mice requires MYD88-dependent signalling and the presence of the microbiota.
Figure 4: Spontaneous development of enteritis and loss of Paneth cells in FADD IEC-KO mice requires RIP3-mediated necrosis of IECs but does not depend on the microbiota.

Similar content being viewed by others

References

  1. Kaser, A., Zeissig, S. & Blumberg, R. S. Inflammatory bowel disease. Annu. Rev. Immunol. 28, 573–621 (2010)

    Article  CAS  Google Scholar 

  2. Strober, W., Fuss, I. & Mannon, P. The fundamental basis of inflammatory bowel disease. J. Clin. Invest. 117, 514–521 (2007)

    Article  CAS  Google Scholar 

  3. MacDonald, T. T. & Monteleone, G. Immunity, inflammation, and allergy in the gut. Science 307, 1920–1925 (2005)

    Article  ADS  CAS  Google Scholar 

  4. Xavier, R. J. & Podolsky, D. K. Unravelling the pathogenesis of inflammatory bowel disease. Nature 448, 427–434 (2007)

    Article  ADS  CAS  Google Scholar 

  5. Turner, J. R. Intestinal mucosal barrier function in health and disease. Nature Rev. Immunol. 9, 799–809 (2009)

    Article  CAS  Google Scholar 

  6. Wilson, N. S., Dixit, V. & Ashkenazi, A. Death receptor signal transducers: nodes of coordination in immune signaling networks. Nature Immunol. 10, 348–355 (2009)

    Article  CAS  Google Scholar 

  7. Cho, Y. S. et al. Phosphorylation-driven assembly of the RIP1–RIP3 complex regulates programmed necrosis and virus-induced inflammation. Cell 137, 1112–1123 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  8. He, S. et al. Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-α. Cell 137, 1100–1111 (2009)

    Article  CAS  Google Scholar 

  9. Zhang, D. W. et al. RIP3, an energy metabolism regulator that switches TNF-induced cell death from apoptosis to necrosis. Science 325, 332–336 (2009)

    Article  ADS  CAS  Google Scholar 

  10. Hitomi, J. et al. Identification of a molecular signaling network that regulates a cellular necrotic cell death pathway. Cell 135, 1311–1323 (2008)

    Article  CAS  Google Scholar 

  11. Nenci, A. et al. Epithelial NEMO links innate immunity to chronic intestinal inflammation. Nature 446, 557–561 (2007)

    Article  ADS  CAS  Google Scholar 

  12. Zhang, J., Cado, D., Chen, A., Kabra, N. H. & Winoto, A. Fas-mediated apoptosis and activation-induced T-cell proliferation are defective in mice lacking FADD/Mort1. Nature 392, 296–300 (1998)

    Article  ADS  CAS  Google Scholar 

  13. Yeh, W. C. et al. FADD: essential for embryo development and signaling from some, but not all, inducers of apoptosis. Science 279, 1954–1958 (1998)

    Article  ADS  CAS  Google Scholar 

  14. Vandenabeele, P., Galluzzi, L., Vanden Berghe, T. & Kroemer, G. Molecular mechanisms of necroptosis: an ordered cellular explosion. Nature Rev. Mol. Cell Biol. 11, 700–714 (2010)

    Article  CAS  Google Scholar 

  15. Holler, N. et al. Fas triggers an alternative, caspase-8-independent cell death pathway using the kinase RIP as effector molecule. Nature Immunol. 1, 489–495 (2000)

    Article  ADS  CAS  Google Scholar 

  16. Osborn, S. L. et al. Fas-associated death domain (FADD) is a negative regulator of T-cell receptor-mediated necroptosis. Proc. Natl Acad. Sci. USA 107, 13034–13039 (2010)

    Article  ADS  CAS  Google Scholar 

  17. Upton, J. W., Kaiser, W. J. & Mocarski, E. S. Virus inhibition of RIP3-dependent necrosis. Cell Host Microbe 7, 302–313 (2010)

    Article  CAS  Google Scholar 

  18. Newton, K., Sun, X. & Dixit, V. M. Kinase RIP3 is dispensable for normal NF-κBs, signaling by the B-cell and T-cell receptors, tumor necrosis factor receptor 1, and Toll-like receptors 2 and 4. Mol. Cell. Biol. 24, 1464–1469 (2004)

    Article  CAS  Google Scholar 

  19. Kovalenko, A. et al. The tumour suppressor CYLD negatively regulates NF-κB signalling by deubiquitination. Nature 424, 801–805 (2003)

    Article  ADS  CAS  Google Scholar 

  20. Wang, L., Du, F. & Wang, X. TNF-α induces two distinct caspase-8 activation pathways. Cell 133, 693–703 (2008)

    Article  CAS  Google Scholar 

  21. Kollias, G. TNF pathophysiology in murine models of chronic inflammation and autoimmunity. Semin. Arthritis Rheum. 34, 3–6 (2005)

    Article  CAS  Google Scholar 

  22. Peyrin-Biroulet, L. Anti-TNF therapy in inflammatory bowel diseases: a huge review. Minerva Gastroenterol. Dietol. 56, 233–243 (2010)

    CAS  PubMed  Google Scholar 

  23. Kaser, A. et al. XBP1 links ER stress to intestinal inflammation and confers genetic risk for human inflammatory bowel disease. Cell 134, 743–756 (2008)

    Article  CAS  Google Scholar 

  24. Cadwell, K. et al. A key role for autophagy and the autophagy gene Atg16l1 in mouse and human intestinal Paneth cells. Nature 456, 259–263 (2008)

    Article  ADS  CAS  Google Scholar 

  25. Dourmashiin, R. R. et al. Epithelial patchy necrosis in Crohn’s disease. Hum. Pathol. 14, 643–648 (1983)

    Article  Google Scholar 

  26. Luedde, T. et al. Deletion of NEMO/IKKγ in liver parenchymal cells causes steatohepatitis and hepatocellular carcinoma. Cancer Cell 11, 119–132 (2007)

    Article  CAS  Google Scholar 

  27. Mc Guire, C. et al. Oligodendrocyte-specific FADD deletion protects mice from autoimmune-mediated demyelination. J. Immunol. 185, 7646–7653 (2010)

    Article  CAS  Google Scholar 

  28. Zhang, H. et al. Functional complementation between FADD and RIP1 in embryos and lymphocytes. Nature 471, 373–376 (2011)

    Article  ADS  CAS  Google Scholar 

  29. Oberst, A. et al. Catalytic activity of the caspase-8–FLIPL complex inhibits RIPK3-dependent necrosis. Nature 471, 363–367 (2011)

    Article  ADS  CAS  Google Scholar 

  30. Kaiser, W. J. et al. RIP3 mediates the embryonic lethality of caspase-8-deficient mice. Nature 471, 368–372 (2011)

    Article  ADS  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. Pasparakis, M., Alexopoulou, L., Episkopou, V. & Kollias, G. Immune and inflammatory responses in TNFα-deficient mice: a critical requirement for TNFα in the formation of primary B cell follicles, follicular dendritic cell networks and germinal centers, and in the maturation of the humoral immune response. J. Exp. Med. 184, 1397–1411 (1996)

    Article  CAS  Google Scholar 

  33. Adachi, O. et al. Targeted disruption of the MyD88 gene results in loss of IL-1- and IL-18-mediated function. Immunity 9, 143–150 (1998)

    Article  CAS  Google Scholar 

  34. Becker, C. et al. In vivo imaging of colitis and colon cancer development in mice using high resolution chromoendoscopy. Gut 54, 950–954 (2005)

    Article  CAS  Google Scholar 

  35. Ukena, S. N. et al. Probiotic Escherichia coli Nissle 1917 inhibits leaky gut by enhancing mucosal integrity. PLoS ONE 2, e1308 (2007)

    Article  ADS  Google Scholar 

  36. Schmidt-Supprian, M. et al. NEMO/IKKγ-deficient mice model incontinentia pigmenti. Mol. Cell 5, 981–992 (2000)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank C. Uthoff-Hachenberg, J. Pfeiffer, E. Mahlberg, D. Beier, J. Buchholz, B. Huelser, B. Wolff, E. Merkel and S. Schmidt for technical support. We also thank V. Dixit and K. Newton for providing Ripk3−/− mice and R. Massoumi for providing anti-CYLD antibody. This work was funded by grants from the Deutsche Forschungsgemeinschaft (SFB 670, SFB 829, CECAD) and European Commission FP7 program grants ‘INFLA-CARE’ and ‘Masterswitch’ (EC contract numbers 223151 and 223404 respectively) to M.P. V. F.-M. was supported by a long-term EMBO fellowship, G.v.L. was supported by ‘Group-ID MRP’ of Ghent University and P.-S.W. was supported by a fellowship from the International Graduate School in Genetics and Functional Genomics at the University of Cologne.

Author information

Authors and Affiliations

Authors

Contributions

P.-S.W., A.W., K.V., V.K., V.F.-M., M.E., P.K. and G.v.L. performed the research. P.-S.W., A.W., K.V., V.K., V.F.-M. and A.S.-K. analysed the data and contributed to writing the manuscript. M.P. provided ideas, co-ordinated the project and wrote the manuscript.

Corresponding author

Correspondence to Manolis Pasparakis.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Table 1 and Supplementary Figures 1-9 with legends. (PDF 10650 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Welz, PS., Wullaert, A., Vlantis, K. et al. FADD prevents RIP3-mediated epithelial cell necrosis and chronic intestinal inflammation. Nature 477, 330–334 (2011). https://doi.org/10.1038/nature10273

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature10273

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