Abstract
Susceptibility to Crohn’s disease, a complex inflammatory disease involving the small intestine, is controlled by over 30 loci1. One Crohn’s disease risk allele is in ATG16L1, a gene homologous to the essential yeast autophagy gene ATG16 (ref. 2). It is not known how ATG16L1 or autophagy contributes to intestinal biology or Crohn’s disease pathogenesis. To address these questions, we generated and characterized mice that are hypomorphic for ATG16L1 protein expression, and validated conclusions on the basis of studies in these mice by analysing intestinal tissues that we collected from Crohn’s disease patients carrying the Crohn’s disease risk allele of ATG16L1. Here we show that ATG16L1 is a bona fide autophagy protein. Within the ileal epithelium, both ATG16L1 and a second essential autophagy protein ATG5 are selectively important for the biology of the Paneth cell, a specialized epithelial cell that functions in part by secretion of granule contents containing antimicrobial peptides and other proteins that alter the intestinal environment3. ATG16L1- and ATG5-deficient Paneth cells exhibited notable abnormalities in the granule exocytosis pathway. In addition, transcriptional analysis revealed an unexpected gain of function specific to ATG16L1-deficient Paneth cells including increased expression of genes involved in peroxisome proliferator-activated receptor (PPAR) signalling and lipid metabolism, of acute phase reactants and of two adipocytokines, leptin and adiponectin, known to directly influence intestinal injury responses. Importantly, Crohn’s disease patients homozygous for the ATG16L1 Crohn’s disease risk allele displayed Paneth cell granule abnormalities similar to those observed in autophagy-protein-deficient mice and expressed increased levels of leptin protein. Thus, ATG16L1, and probably the process of autophagy, have a role within the intestinal epithelium of mice and Crohn’s disease patients by selective effects on the cell biology and specialized regulatory properties of Paneth cells.
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Acknowledgements
We thank N. Abe for technical assistance with fluorescence microscopy, V. Cavalli for microscope use, J. Eisenberg and A. Ng for help with bioinformatics analysis, A. J. Ouellette for technical advice, and I. Mysorekar and J. Mills for help with antibodies. This research was supported by grant U54 AI057160 Project 6 and the Broad Foundation (K.C., J. Loh and H.W.V.), training grant NIH T32 AR07279 (K.C.), the Lallage Feazel Wall Fellowship DRG-1972-08 (K.C.) from the Damon Runyon Cancer Research Foundation; the Pew Foundation (J. Liu, S.L.B., H.M., W.K. and T.S.S.); the Washington University Digestive Diseases Research Core Center P30 DK52574, Barnes Jewish Foundation, Johnson and Johnson Translational Seed Award, and the Crohn’s Colitis Foundation of America (E.L., S.H. and C.S.); grants-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and the Toray Science Foundation (C.K. and N.M.); NIH R01 AI062832 (J.A.C.); and NIH AI062773 and DK43351 (R.J.X.).
Author Contributions The original hypothesis of the article was formulated by K.C., H.W.V. and T.S.S. Autophagy experiments were performed by K.C. and C.K. Mouse lines were established by K.C., J. Loh and B.P.S. Mouse analysis experiments were performed by K.C., J. Liu, S.L.B., H.M., W.K., T.S.S. and J. Lennerz. Histological examination was performed by T.S.S, E.M.B. and J. Lennerz. L. monocytogenes experiments were performed by J.C. and K.C. Microarray analysis was performed by R.J.X. and T.S.S. Human tissue collection, genotyping and preparation were performed by E.L., S.H. and C.S. N.M. provided advice, the antibody to ATG16L1, and experiments by C.K. were performed in N.M.’s laboratory. The manuscript was written by K.C., H.W.V. and T.S.S., and all authors commented on the manuscript, data and conclusions before submission.
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Supplementary Table 1. Microarray analysis of RNA from crypt-base epithelial cells (XLS 58 kb)
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Cadwell, K., Liu, J., Brown, S. et al. A key role for autophagy and the autophagy gene Atg16l1 in mouse and human intestinal Paneth cells. Nature 456, 259–263 (2008). https://doi.org/10.1038/nature07416
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DOI: https://doi.org/10.1038/nature07416
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