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Caspase-8 regulates TNF-α-induced epithelial necroptosis and terminal ileitis

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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Figure 1: Casp8 ΔIEC mice spontaneously develop ileitis and lack Paneth cells.
Figure 2: Increased caspase-8-independent cell death within crypts of Casp8 ΔIEC mice.
Figure 3: Inhibition of TNF-α-induced epithelial necroptosis in Casp8 ΔIEC mice.
Figure 4: RIP-mediated necroptosis of Paneth cells in patients with Crohn’s disease.

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Gene Expression Omnibus

Data deposits

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).

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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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Correspondence to Christoph Becker.

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The authors declare no competing financial interests.

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Supplementary Information

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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