PMID- 19786028 OWN - NLM STAT- MEDLINE DCOM- 20100315 LR - 20220318 IS - 1528-0012 (Electronic) IS - 0016-5085 (Print) IS - 0016-5085 (Linking) VI - 138 IP - 1 DP - 2010 Jan TI - Toll-like receptor-4 inhibits enterocyte proliferation via impaired beta-catenin signaling in necrotizing enterocolitis. PG - 185-96 LID - 10.1053/j.gastro.2009.09.045 [doi] AB - BACKGROUND & AIMS: Necrotizing enterocolitis (NEC), the leading cause of gastrointestinal death from gastrointestinal disease in preterm infants, is characterized by exaggerated TLR4 signaling and decreased enterocyte proliferation through unknown mechanisms. Given the importance of beta-catenin in regulating proliferation of many cell types, we hypothesize that TLR4 impairs enterocyte proliferation in NEC via impaired beta-catenin signaling. METHODS: Enterocyte proliferation was detected in IEC-6 cells or in ileum or colon from wild-type, TLR4-mutant, or TLR4(-/-) mice after induction of NEC or endotoxemia. beta-Catenin signaling was assessed by cell fractionation or immunoconfocal microscopy to detect its nuclear translocation. Activation and inhibition of beta-catenin were achieved via cDNA or small interfering RNA, respectively. TLR4 in the intestinal mucosa was inhibited with adenoviruses expressing dominant-negative TLR4. RESULTS: TLR4 activation significantly impaired enterocyte proliferation in the ileum but not colon in newborn but not adult mice and in IEC-6 enterocytes. beta-Catenin activation reversed these effects in vitro. To determine the mechanisms involved, TLR4 activation phosphorylated the upstream inhibitory kinase GSK3beta, causing beta-catenin degradation. NEC in both mouse and humans was associated with decreased beta-catenin and increased mucosal GSK3beta expression. Strikingly, the inhibition of enterocyte beta-catenin signaling in NEC could be reversed, and enterocyte proliferation restored, through adenoviral-mediated inhibition of TLR4 signaling in the small intestinal mucosa. CONCLUSION: We now report a novel pathway linking TLR4 with inhibition of beta-catenin signaling via GSK3beta activation, leading to reduced enterocyte proliferation in vitro and in vivo. These data provide additional insights into the pathogenesis of diseases of intestinal inflammation such as NEC. CI - Copyright 2010 AGA Institute. Published by Elsevier Inc. All rights reserved. FAU - Sodhi, Chhinder P AU - Sodhi CP AD - Division of Pediatric Surgery, Department of Surgery, Children's Hospital of Pittsburgh and University of Pittsburgh, Pittsburgh, Pennsylvania 15224, USA. FAU - Shi, Xia-Hua AU - Shi XH FAU - Richardson, Ward M AU - Richardson WM FAU - Grant, Zachary S AU - Grant ZS FAU - Shapiro, Richard A AU - Shapiro RA FAU - Prindle, Thomas Jr AU - Prindle T Jr FAU - Branca, Maria AU - Branca M FAU - Russo, Anthony AU - Russo A FAU - Gribar, Steven C AU - Gribar SC FAU - Ma, Congrong AU - Ma C FAU - Hackam, David J AU - Hackam DJ LA - eng GR - R01 GM078238-01/GM/NIGMS NIH HHS/United States GR - R01DK08752/DK/NIDDK NIH HHS/United States GR - R01 DK083752/DK/NIDDK NIH HHS/United States GR - R01 GM078238/GM/NIGMS NIH HHS/United States GR - R01GM078238/GM/NIGMS NIH HHS/United States GR - R01 DK083752-02/DK/NIDDK NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20090926 PL - United States TA - Gastroenterology JT - Gastroenterology JID - 0374630 RN - 0 (CTNNB1 protein, mouse) RN - 0 (Lipopolysaccharides) RN - 0 (Tlr4 protein, mouse) RN - 0 (Toll-Like Receptor 4) RN - 0 (beta Catenin) RN - 0 (lipopolysaccharide A) RN - EC 2.7.11.1 (GSK3B protein, human) RN - EC 2.7.11.1 (Glycogen Synthase Kinase 3 beta) RN - EC 2.7.11.1 (Gsk3b protein, mouse) RN - EC 2.7.11.26 (Glycogen Synthase Kinase 3) SB - IM CIN - Gastroenterology. 2010 Jan;138(1):39-43. PMID: 19932667 MH - Adenoviridae/genetics MH - Animals MH - Cell Division/drug effects/physiology MH - Cells, Cultured MH - Colon/pathology MH - Enterocolitis, Necrotizing/*metabolism/pathology MH - Enterocytes/*cytology/*metabolism MH - Glycogen Synthase Kinase 3/metabolism MH - Glycogen Synthase Kinase 3 beta MH - Humans MH - Ileum/pathology MH - Infant, Newborn MH - Intestinal Mucosa/metabolism/pathology MH - Lipopolysaccharides/pharmacology MH - Mice MH - Mice, Inbred C3H MH - Mice, Mutant Strains MH - Signal Transduction/physiology MH - Toll-Like Receptor 4/genetics/*metabolism MH - beta Catenin/*metabolism PMC - PMC2813409 MID - NIHMS150379 COIS- Conflicts of interest: Xia-hua Shi - no conflicts exist; Chhinder P. Sodhi - - no conflicts exist ; Ward M. Richardson - - no conflicts exist; Steven C. Gribar - - no conflicts exist; Thomas Prindle Jr - - no conflicts exist; Maria Branca - no conflicts exist; Anthony Russo - - no conflicts exist; Congrong Ma - - no conflicts exist ; Richard Shapiro - no conflicts exist; Zachary Grant - no conflicts exist; David J. Hackam - no conflicts exist. Transcript profiling - Not applicable Writing assistance - Not applicable. EDAT- 2009/09/30 06:00 MHDA- 2010/03/17 06:00 PMCR- 2011/01/01 CRDT- 2009/09/30 06:00 PHST- 2009/04/16 00:00 [received] PHST- 2009/08/19 00:00 [revised] PHST- 2009/09/20 00:00 [accepted] PHST- 2009/09/30 06:00 [entrez] PHST- 2009/09/30 06:00 [pubmed] PHST- 2010/03/17 06:00 [medline] PHST- 2011/01/01 00:00 [pmc-release] AID - S0016-5085(09)01695-3 [pii] AID - 10.1053/j.gastro.2009.09.045 [doi] PST - ppublish SO - Gastroenterology. 2010 Jan;138(1):185-96. doi: 10.1053/j.gastro.2009.09.045. Epub 2009 Sep 26.