Role of nuclear bile acid receptor FXR in intestinal bacterial overgrowth and translocation
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1 Role of nuclear bile acid receptor FXR in intestinal bacterial overgrowth and translocation Antonio Moschetta University of Bari, Consorzio Mario Negri Sud, Santa Maria Imbaro (Chieti), Italy
2 Plan Bile acids and the gut microbiota Nuclear bile acid receptor FXR FXR and mucosal defense Regulation of intestinal FXR
3 Physiologic Functions of Bile Acids CO 2 H OH CO 2 H HO OH H CDCA HO H CA OH liver Absorption of lipids Elimination of cholesterol gall bladder stomach small intestine ileum
4 Bile Acids Protect Intestine Biliary obstruction in rats causes bacterial overgrowth and translocation, which are reversed by oral bile acids (Ding et al., Eur Surg Res 25: 11; Lorenzo-Zuniga et al., Hepatology 37: 551) In humans, biliary obstruction increases susceptibility to bacterial overgrowth and sepsis Preoperative administration of sodium deoxycholate prevents postoperative endotoxaemia in patients with obstructive jaundice (Cahill et al., Surg Gynecol Obstet 165:519) Does bile nuclear receptor FXR protect against bacterial overgrowth and translocation?
5 Bile Acid Receptor, FXR Highly expressed in liver and intestine Binds DNA to IR-1 motifs as RXR heterodimer Activated by primary bile acids FXR-KO mice have elevated CYP7A1 and bile acid synthesis
6 FXR Target Genes in Ileum Fold Gene +83 fibroblast growth factor orphan nuclear receptor SHP +4. inducible nitric oxide synthase +2.3 angiogenin +2.2 interleukin ileal bile acid binding protein -3.7 MHC Q1 BA homeostasis Mucosal defense
7 Bile Duct Ligation Experiments Day 1: Begin treatment with vehicle or GW464 liver Day 3: Perform or sham operation Day 8: Kill mice and quantitate bacteria in ileum, cecum, and mesenteric lymph nodes gall bladder ileum cecum
8 GW464 Blocks Bacterial Overgrowth in presence of FXR wild type FXRKO 3 4 ileum CFU/ml (x 1 7 ) 2 1 a b a CFU/ml (x 1 7 ) Sham GW464 Sham GW cecum CFU/g (x 1 8 ) 8 4 a Sham b a GW464 CFU/g (x 1 8 ) Sham GW464
9 anti-occludin immunostaining FXR Promotes Barrier Integrity Wild type FXR-KO Sham GW464 Sham
10 FXR-KO Sham Bacterial Translocation in FXR-KO
11 Why Link FXR to Mucosal Defense? 1. Bile acids are strong detergents FXR serves as a rheostat to prevent damage to the mucosal barrier 2. Bile acids are released into the intestine after a meal; since food is an important source of microbes, FXR ensures that appropriate levels of antimicrobial proteins are produced inos IL18 angiogenin mucosal protection inflammatory disease FXR agonists may be useful to treat intestinal disease
12 Intestinal Mucosa Architecture Villus Surface Epithelium Differentiation Differentiation Proliferative progenitors Crypt Proliferative progenitors Crypt Stem cells Stem cells Ileum Colon Clevers H. Cell 26
13 FXR expression increases during crypt-to-villus differentiation merge Anti-FXR DAPI protein Reaching confluence
14 FXR expression increases during crypt-to-villus differentiation 12 1 FXRα1/α2 pre-confluence confluence post-confluence mrna
15 FXR disappears in APCm-driven tumors APC min/+ mice FAP patients normal tumor P1 P2 P3 P4 P5 FXRα1/α2 P1 P2 P3 P4 P5 FXRα3/α4
16 Induction of WT APC via tet-on system in APCmHT29 restores FXR expression 1 FXRα1/α ZnCl2 +ZnCl2 4 2
17 Thanks Salvatore Modica, PhD student Giuseppe Losasso, PhD student Michele Petruzzelli, PhD student Ilenia D Errico, Postdoc fellow Stefania Murzilli, Research Tech Lorena Salvatore, Research Tech Filomena Cinalli, Lab Manager Jean-Noel Freund, Strasbourg, Fr Ken Kinzler & Bert Vogelstein, Baltimore US Davo Mango and Steven Kliewer Center, Dallas US Giuseppe Palasciano & Piero Portincasa, Bari, It Italian Association for Cancer Research, University of Bari, EU
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