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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.

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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.

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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.

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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.

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Correspondence to Manolis Pasparakis.

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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

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