Abstract
Dysfunction of the intestinal epithelium is believed to result in the excessive translocation of commensal bacteria into the bowel wall that drives chronic mucosal inflammation in Crohn’s disease, an incurable inflammatory bowel disease in humans characterized by inflammation of the terminal ileum1. In healthy individuals, the intestinal epithelium maintains a physical barrier, established by the tight contact of cells. Moreover, specialized epithelial cells such as Paneth cells and goblet cells provide innate immune defence functions by secreting mucus and antimicrobial peptides, which hamper access and survival of bacteria adjacent to the epithelium2. Epithelial cell death is a hallmark of intestinal inflammation and has been discussed as a possible pathogenic mechanism driving Crohn’s disease in humans3. However, the regulation of epithelial cell death and its role in intestinal homeostasis remain poorly understood. Here we demonstrate a critical role for caspase-8 in regulating necroptosis of intestinal epithelial cells (IECs) and terminal ileitis. Mice with a conditional deletion of caspase-8 in the intestinal epithelium (Casp8ΔIEC) spontaneously developed inflammatory lesions in the terminal ileum and were highly susceptible to colitis. Casp8ΔIEC mice lacked Paneth cells and showed reduced numbers of goblet cells, indicating dysregulated antimicrobial immune cell functions of the intestinal epithelium. Casp8ΔIEC mice showed increased cell death in the Paneth cell area of small intestinal crypts. Epithelial cell death was induced by tumour necrosis factor (TNF)-α, was associated with increased expression of receptor-interacting protein 3 (Rip3; also known as Ripk3) and could be inhibited on blockade of necroptosis. Lastly, we identified high levels of RIP3 in human Paneth cells and increased necroptosis in the terminal ileum of patients with Crohn’s disease, suggesting a potential role of necroptosis in the pathogenesis of this disease. Together, our data demonstrate a critical function of caspase-8 in regulating intestinal homeostasis and in protecting IECs from TNF-α-induced necroptotic cell death.
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Gene Expression Omnibus
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Chip data were deposited at the NCBI Gene Expression Omnibus under the series accession number GSE30873 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE30873).
References
Strober, W., Fuss, I. & Mannon, P. The fundamental basis of inflammatory bowel disease. J. Clin. Invest. 117, 514–521 (2007)
Artis, D. Epithelial-cell recognition of commensal bacteria and maintenance of immune homeostasis in the gut. Nature Rev. Immunol. 8, 411–420 (2008)
Kaser, A., Zeissig, S. & Blumberg, R. S. Inflammatory bowel disease. Annu. Rev. Immunol. 28, 573–621 (2010)
Hall, P. A. et al. Regulation of cell number in the mammalian gastrointestinal tract: the importance of apoptosis. J. Cell Sci. 107, 3569–3577 (1994)
Nenci, A. et al. Epithelial NEMO links innate immunity to chronic intestinal inflammation. Nature 446, 557–561 (2007)
Sanders, D. S. Mucosal integrity and barrier function in the pathogenesis of early lesions in Crohn’s disease. J. Clin. Pathol. 58, 568–572 (2005)
Elphick, D. A. & Mahida, Y. R. Paneth cells: their role in innate immunity and inflammatory disease. Gut 54, 1802–1809 (2005)
Maelfait, J. & Beyaert, R. Non-apoptotic functions of caspase-8. Biochem. Pharmacol. 76, 1365–1373 (2008)
Valmiki, M. G. & Ramos, J. W. Death effector domain-containing proteins. Cell. Mol. Life Sci. 66, 814–830 (2009)
Mielgo, A. et al. The death effector domains of caspase-8 induce terminal differentiation. PLoS ONE 4, e7879 (2009)
Sato, T. et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459, 262–265 (2009)
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)
Vercammen, D. et al. Tumour necrosis factor-induced necrosis versus anti-Fas-induced apoptosis in L929 cells. Cytokine 9, 801–808 (1997)
Vandenabeele, P. et al. Molecular mechanisms of necroptosis: an ordered cellular explosion. Nature Rev. Mol. Cell Biol. 11, 700–714 (2010)
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)
Declercq, W., Vanden Berghe, T. & Vandenabeele, P. RIP kinases at the crossroads of cell death and survival. Cell 138, 229–232 (2009)
He, S. et al. Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-α. Cell 137, 1100–1111 (2009)
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)
Kaiser, W. J. et al. RIP3 mediates the embryonic lethality of caspase-8-deficient mice. Nature 471, 368–372 (2011)
Oberst, A. et al. Catalytic activity of the caspase-8–FLIPL complex inhibits RIPK3-dependent necrosis. Nature 471, 363–367 (2011)
Degterev, A. et al. Identification of RIP1 kinase as a specific cellular target of necrostatins. Nature Chem. Biol. 4, 313–321 (2008)
Wehkamp, J. et al. Barrier dysfunction due to distinct defensin deficiencies in small intestinal and colonic Crohn’s disease. Mucosal Immunol. 1 (Suppl. 1). S67–S74 (2008)
Lewin, K. The Paneth cell in disease. Gut 10, 804–811 (1969)
Arijs, I. et al. Mucosal gene expression of antimicrobial peptides in inflammatory bowel disease before and after first infliximab treatment. PLoS ONE 4, e7984 (2009)
Mayhew, T. M. et al. Epithelial integrity, cell death and cell loss in mammalian small intestine. Histol. Histopathol. 14, 257–267 (1999)
Kontoyiannis, D. et al. Impaired on/off regulation of TNF biosynthesis in mice lacking TNF AU-rich elements: implications for joint and gut-associated immunopathologies. Immunity 10, 387–398 (1999)
Beisner, D. R. et al. Cutting edge: innate immunity conferred by B cells is regulated by caspase-8. J. Immunol. 175, 3469–3473 (2005)
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)
El Marjou, F. et al. Tissue-specific and inducible Cre-mediated recombination in the gut epithelium. Genesis 39, 186–193 (2004)
Becker, C. et al. In vivo imaging of colitis and colon cancer development in mice using high resolution chromoendoscopy. Gut 54, 950–954 (2005)
Acknowledgements
The research leading to these results has received funding from the Interdisciplinary Center for Clinical Research (IZKF) of the University Erlangen-Nuremberg and the European Community's 7th Framework Program (FP7/2007-2013) under grant agreement no. 202230, acronym GENINCA. E.M. received funding from the Welcome Trust (WT087768MA) and S.M.H. was supported by NIH grant AI037988. The authors thank A. Watson for critical reading of the manuscript, A. Nikolaev, S. Wallmüller, V. Buchert and M. Klewer for technical assistance and J. Mudter, R. Atreya and C. Neufert for sampling biopsies.
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C.G., K.A., M.F.N. and C.B. designed the research. C.G., E.M., N.W., B.W., H.N., M.W. and S.T. performed the experiments. S.M.H. provided material that made the study possible. C.G., K.A. and C.B. analysed the data and wrote the paper.
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This file contains Supplementary Figures 1-11 with legends and Supplementary Table 1 displaying the results of gene chip experiments comparing control and Casp8ΔIEC mice. (PDF 1663 kb)
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Günther, C., Martini, E., Wittkopf, N. et al. Caspase-8 regulates TNF-α-induced epithelial necroptosis and terminal ileitis. Nature 477, 335–339 (2011). https://doi.org/10.1038/nature10400
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DOI: https://doi.org/10.1038/nature10400
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