Center for Molecular Medicine Cologne

Pasparakis, Manolis - B 07

Role of FADD and caspase-8 in the regulation of intestinal epithelial cell death and inflammation


Inflammatory bowel diseases (IBD) are chronic relapsing inflammatory disorders of the gastrointestinal tract with increasing incidence over the past 50 years. Despite a wide variety of possible triggers, inappropriate immune responses to the commensal microbiota are generally considered a central underlying theme in IBD. The intestinal epithelium forms a structural barrier between the commensal bacteria residing in the intestinal lumen and the mucosal immune cells and plays a critical role for the maintenance of gut immune homeostasis.

We showed previously that mice lacking FADD in intestinal epithelial cells (FADDIEC-KO) develop chronic ileitis and colitis due to RIPK3-mediated death of IECs. Moreover, mice with IEC-specific knockout of caspase-8 (Casp8IEC‑KO) were reported to develop ileitis but not colitis, suggesting that FADD and caspase-8 may have different functions in the regulation of intestinal epithelial homeostasis and inflammation.

Here we aim to dissect the mechanisms by which FADD and caspase-8 regulate intestinal inflammation using genetic mouse models. The recent discovery that human patients with caspase-8 mutations develop severe intestinal inflammation suggests that the FADD/Casp8-dependent pathways regulating intestinal homeostasis and inflammation are conserved between humans and mice.

Therefore, our project is relevant for the pathogenesis of intestinal inflammation in humans and may lead to the identification of new therapeutic targets for patients with IBD.

Our Aims

  1. Dissect the upstream pathways that regulate the development of colitis and ileitis in FADDIEC-KO and CASP8IEC-KO  mice

  2. Unravel the mechanisms by which FADD or Caspase-8 act in intestinal epithelial cells to regulate immune homeostasis and prevent mucosal inflammation

  3. Study the interplay of FADD/Casp8 with other pathways that are important for the pathogenesis of inflammatory bowel diseases

Previous Work

Impaired epithelial barrier function was reported in IBD patients and their healthy first-degree relatives, identifying epithelial permeability defects as possible causal factors in IBD. Our previous studies in mouse models provided evidence that death of intestinal epithelial cells (IECs) triggers severe chronic intestinal inflammation by disrupting the epithelial barrier.

We showed that mice lacking nuclear factor κB (NF-κB) essential modulator (NEMO, also named IKKγ) specifically in intestinal epithelial cells (IECs) develop chronic inflammatory colitis and ileitis that is induced by FADD/RIPK3-dependent death of IECs and depends on the presence of the intestinal microbiota (Nenci et al., 2007; Vlantis et al., 2016). In addition, we showed previously that mice with IEC-specific FADD deficiency developed severe chronic inflammatory colitis that depended on the presence of the intestinal microbiota and TNF (Welz et al., 2011).

Moreover, FADDIEC-KO mice also developed ileitis with loss of Paneth cells, secretory cells that reside in the intestinal crypt and are believed to have important functions in the regulation of the stem cell niche and the crypt microbiota. Interestingly, when FADDIEC-KO mice were raised under germ-free conditions they did not develop colitis but developed ileitis, suggesting that different mechanisms regulate the inflammatory response in the large and small intestine. Furthermore, TNF deficiency inhibited colitis but did not prevent ileitis also pointing towards a fundamentally different mechanism triggering the disease in the colon and the ileum.

However, RIPK3 deficiency prevented both ileitis and colitis, suggesting that FADD prevents inflammation in both the small and large intestine by preventing epithelial cell necroptosis (Welz et al., 2011).

  1. Nenci, A., Becker, C., Wullaert, A., Gareus, R., van Loo, G., Danese, S., Huth, M., Nikolaev, A., Neufert, C., Madison, B., Gumucio, D., Neurath, M.F., and Pasparakis, M. (2007). Epithelial NEMO links innate immunity to chronic intestinal inflammation. Nature 446, 557-561.
  2. Vlantis, K., Wullaert, A., Polykratis, A., Kondylis, V., Dannappel, M., Schwarzer, R., Welz, P., Corona, T., Walczak, H., Weih, F., Klein, U., Kelliher, M., and Pasparakis, M. (2016). NEMO Prevents RIP Kinase 1-Mediated Epithelial Cell Death and Chronic Intestinal Inflammation by NF-kappaB-Dependent and -Independent Functions. Immunity 44, 553-567.
  3. Welz, P.S., Wullaert, A., Vlantis, K., Kondylis, V., Fernandez-Majada, V., Ermolaeva, M., Kirsch, P., Sterner-Kock, A., van Loo, G., and Pasparakis, M. (2011). FADD prevents RIP3-mediated epithelial cell necrosis and chronic intestinal inflammation. Nature 477, 330-334.
  • Kumari S, Van TM, Preukschat D, Schuenke H, Basic M, Bleich A, Klein U, and Pasparakis M (2021). NF-kappaB inhibition in keratinocytes causes RIPK1-mediated necroptosis and skin inflammation. Life Sci Alliance4. doi:10.26508/lsa.202000956.
  • Oikonomou N, Schuijs MJ, Chatzigiagkos A, Androulidaki A, Aidinis V, Hammad H, Lambrecht BN, and Pasparakis M (2021). Airway epithelial cell necroptosis contributes to asthma exacerbation in a mouse model of house dust mite-induced allergic inflammation. Mucosal Immunol14, 1160-1171. doi:10.1038/s41385-021-00415-5.
                                                                                                                            • Schunke H, Gobel U, Dikic I, and Pasparakis M (2021). OTULIN inhibits RIPK1-mediated keratinocyte necroptosis to prevent skin inflammation in mice. Nat Commun12, 5912. doi:10.1038/s41467-021-25945-1.
                                                                                                                                                                          • Theobald SJ, Grab J, Fritsch M, Suarez I, Eisfeld HS, Winter S, Koch M, Holscher C, Pasparakis M, Kashkar H, and Rybniker J (2021). Gasdermin D mediates host cell death but not interleukin-1beta secretion in Mycobacterium tuberculosis-infected macrophages. Cell Death Discov7, 327. doi:10.1038/s41420-021-00716-5.
                                                                                                                                                                          • Jiao H, Wachsmuth L, Kumari S, Schwarzer R, Lin J, Eren RO, Fisher A, Lane R, Young GR, Kassiotis G, Kaiser WJ, and Pasparakis M (2020b). Z-nucleic-acid sensing triggers ZBP1-dependent necroptosis and inflammation. Nature 580, 391-5.
                                                                                                                                                                          • Lalaoui N, Boyden SE, Oda H, Wood GM, Stone DL, Chau D, Liu L, Stoffels M, Kratina T, Lawlor KE, Zaal KJM, Hoffmann PM, Etemadi N, Shield-Artin K, Biben C, Tsai WL, Blake MD, Kuehn HS, Yang D, Anderton H, Silke N, Wachsmuth L, Zheng L, Moura NS, Beck DB, Gutierrez-Cruz G, Ombrello AK, Pinto-Patarroyo GP, Kueh AJ, Herold MJ, Hall C, Wang H, Chae JJ, Dmitrieva NI, McKenzie M, Light A, Barham BK, Jones A, Romeo TM, Zhou Q, Aksentijevich I, Mullikin JC, Gross AJ, Shum AK, Hawkins ED, Masters SL, Lenardo MJ, Boehm M, Rosenzweig SD, Pasparakis M, Voss AK, Gadina M, Kastner DL, and Silke J (2020). Mutations that prevent caspase cleavage of RIPK1 cause autoinflammatory disease. Nature 577, 103-8.
                                                                                                                                                                          • Laurien L, Nagata M, Schunke H, Delanghe T, Wiederstein JL, Kumari S, Schwarzer R, Corona T, Kruger M, Bertrand MJM, Kondylis V, and Pasparakis M (2020). Autophosphorylation at serine 166 regulates RIP kinase 1-mediated cell death and inflammation. Nat Commun 11, 1747.
                                                                                                                                                                          • Schiffmann LM, Werthenbach JP, Heintges-Kleinhofer F, Seeger JM, Fritsch M, Gunther SD, Willenborg S, Brodesser S, Lucas C, Jungst C, Albert MC, Schorn F, Witt A, Moraes CT, Bruns CJ, Pasparakis M, Kronke M, Eming SA, Coutelle O, and Kashkar H (2020). Mitochondrial respiration controls neoangiogenesis during wound healing and tumour growth. Nat Commun 11.
                                                                                                                                                                          • Verboom L, Martens A, Priem D, Hoste E, Sze M, Vikkula H, Van Hove L, Voet S, Roels J, Maelfait J, Bongiovanni L, de Bruin A, Scott CL, Saeys Y, Pasparakis M, Bertrand MJM, and van Loo G (2020). OTULIN Prevents Liver Inflammation and Hepatocellular Carcinoma by Inhibiting FADD- and RIPK1 Kinase-Mediated Hepatocyte Apoptosis. Cell Rep 30, 2237-47 e6.
                                                                                                                                                                          • Wong J, Garcia-Carbonell R, Zelic M, Ho SB, Boland BS, Yao SJ, Desai SA, Das S, Planell N, Harris PA, Font-Burgada J, Taniguchi K, Bertin J, Salas A, Pasparakis M, Gough PJ, Kelliher M, Karin M, and Guma M (2020). RIPK1 Mediates TNF-Induced Intestinal Crypt Apoptosis During Chronic NF-kappaB Activation. Cell Mol Gastroenterol Hepatol 9, 295-312.
                                                                                                                                                                          • Schwarzer R, Laurien L, and Pasparakis M (2020). New insights into the regulation of apoptosis, necroptosis, and pyroptosis by receptor interacting protein kinase 1 and caspase-8. Curr Opin Cell Biol63, 186-193. doi:10.1016/
                                                                                                                                                                          • Schwarzer, R., Jiao, H., Wachsmuth, L., Tresch, A. & Pasparakis, M. FADD and Caspase-8 Regulate Gut Homeostasis and Inflammation by Controlling MLKL- and GSDMD-Mediated Death of Intestinal Epithelial Cells. Immunity 52, 978-993.e6 (2020).
                                                                                                                                                                          Prof. Dr. Manolis Pasparakis CMMC Cologne
                                                                                                                                                                          Prof. Dr. Manolis Pasparakis

                                                                                                                                                                          Institute for Genetics - CECAD Research Center

                                                                                                                                                                          CMMC - PI - B 07

                                                                                                                                                                          +49 221 478 84351

                                                                                                                                                                          +49 221 478 6360

                                                                                                                                                                          Institute for Genetics - CECAD Research Center

                                                                                                                                                                          Joseph-Stelzmann-Str. 26

                                                                                                                                                                          50931 Cologne

                                                                                                                                                                          CMMC Profile Page

                                                                                                                                                                          Curriculum Vitae (CV)

                                                                                                                                                                          Publications on PubMed

                                                                                                                                                                          Publications - Manolis Pasparakis

                                                                                                                                                                          Link to PubMed

                                                                                                                                                                          Figure 1