DUOX2 variants associate with preclinical disturbances in microbiota-immune homeostasis and increased inflammatory bowel disease risk
Helmut Grasberger, … , Gilbert S. Omenn, John Y. Kao
Helmut Grasberger, … , Gilbert S. Omenn, John Y. Kao
Published March 2, 2021
Citation Information: J Clin Invest. 2021;131(9):e141676. https://doi.org/10.1172/JCI141676.
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Research Article Gastroenterology

DUOX2 variants associate with preclinical disturbances in microbiota-immune homeostasis and increased inflammatory bowel disease risk

Abstract

A primordial gut-epithelial innate defense response is the release of hydrogen peroxide by dual NADPH oxidase (DUOX). In inflammatory bowel disease (IBD), a condition characterized by an imbalanced gut microbiota-immune homeostasis, DUOX2 isoenzyme is the highest induced gene. Performing multiomic analyses using 2872 human participants of a wellness program, we detected a substantial burden of rare protein-altering DUOX2 gene variants of unknown physiologic significance. We identified a significant association between these rare loss-of-function variants and increased plasma levels of interleukin-17C, which is induced also in mucosal biopsies of patients with IBD. DUOX2-deficient mice replicated increased IL-17C induction in the intestine, with outlier high Il17c expression linked to the mucosal expansion of specific Proteobacteria pathobionts. Integrated microbiota/host gene expression analyses in patients with IBD corroborated IL-17C as a marker for epithelial activation by gram-negative bacteria. Finally, the impact of DUOX2 variants on IL-17C induction provided a rationale for variant stratification in case control studies that substantiated DUOX2 as an IBD risk gene. Thus, our study identifies an association of deleterious DUOX2 variants with a preclinical hallmark of disturbed microbiota-immune homeostasis that appears to precede the manifestation of IBD.

Authors

Helmut Grasberger, Andrew T. Magis, Elisa Sheng, Matthew P. Conomos, Min Zhang, Lea S. Garzotto, Guoqing Hou, Shrinivas Bishu, Hiroko Nagao-Kitamoto, Mohamad El-Zaatari, Sho Kitamoto, Nobuhiko Kamada, Ryan W. Stidham, Yasutada Akiba, Jonathan Kaunitz, Yael Haberman, Subra Kugathasan, Lee A. Denson, Gilbert S. Omenn, John Y. Kao

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Figure 5

High Il17c expression in the intestinal epithelium of DUOX2-deficient mice is linked to the expansion of gram-negative pathobionts.

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High Il17c expression in the intestinal epithelium of DUOX2-deficient mi...
(A) Differential microbiota-dependent regulation of Il17c, Il17a, and Reg3g (IL-22 target gene) in the mouse intestine. GF, germ-free; CONV, conventionalized (SPF); SFBmono, monocolonized with segmented filamentous bacteria. n = 5 animals per condition. Data represent median expression values with IQR. Kruskal-Wallis with Dunn’s post hoc test. (B) Mice were treated for 3 days with an antibiotics (Abx) regimen comprising ciprofloxacin and metronidazole that suppresses the gram-negative gut microbiota (see Supplemental Figure 4). n = 6 and n = 5 for control mice without or with Abx treatment, respectively, and n = 8 and n = 4 for intestinal epithelial-specific Duoxa–/– mice without or with Abx treatment, respectively. Data represent geometric means with 95% CI. Kruskal-Wallis test with Dunn’s post hoc test. (C) Acute cell-autonomous induction of Il17c expression in enteroid-derived epithelial monolayers directly exposed to bacteria. Each treatment was performed on 6 independent enteroid cultures derived from 3 Duoxa–/–/WT littermate pairs. Bars indicate median expression values. Kruskal-Wallis with Dunn’s post hoc test. (D) Cladogram (phylum to genus level) depicting results of LEfSe (54) analysis identifying taxa with distinct relative abundance (P < 0.01; LDA > 2) in ileal mucosa of Duoxa–/– (n = 26) compared with WT (n = 22) littermates. (E) Discriminative taxa in the ileal mucosal microbiota of Il17chi animals (marked with arrows in Figure 4A). (F) The relative mucosal abundance of genus Helicobacter. Data represent median values with IQR. Two-tailed Mann-Whitney. (G) The relative abundance of Proteobacterium otu0194 vs mucosal Il17c expression. Arrows in F and G indicate animals classified as Il17chi in Figure 4A. *P < 0.05; **P < 0.01; ***P < 0.001.