MicroRNA 29 targets nuclear factor-κB-repressing factor and Claudin 1 to increase intestinal permeability

Gastroenterology. 2015 Jan;148(1):158-169.e8. doi: 10.1053/j.gastro.2014.09.037. Epub 2014 Sep 30.

Abstract

Background & aims: Some patients with irritable bowel syndrome with diarrhea (IBS-D) have intestinal hyperpermeability, which contributes to their diarrhea and abdominal pain. MicroRNA 29 (MIR29) regulates intestinal permeability in patients with IBS-D. We investigated and searched for targets of MIR29 and investigated the effects of disrupting Mir29 in mice.

Methods: We investigated expression MIR29A and B in intestinal biopsies collected during endoscopy from patients with IBS (n = 183) and without IBS (controls) (n = 36). Levels were correlated with disease phenotype. We also generated and studied Mir29(-/-) mice, in which expression of Mir29a and b, but not c, is lost. Colitis was induced by administration of 2,4,6-trinitrobenzenesulfonic acid; intestinal tissues were collected and permeability was assessed. Microarray analysis was performed using tissues from Mir29(-/-) mice. Changes in levels of target genes were measured in human colonic epithelial cells and small intestinal epithelial cells after knockdown of MIR29 with anti-MIRs.

Results: Intestinal tissues from patients with IBS-D (but not IBS with constipation or controls) had increased levels of MIR29A and B, but reduced levels of Claudin-1 (CLDN1) and nuclear factor-κB-repressing factor (NKRF). Induction of colitis and water avoidance stress increased levels of Mir29a and Mir29b and intestinal permeability in wild-type mice; these increased intestinal permeability in colons of far fewer Mir29(-/-) mice. In microarray and knockdown experiments, MIR29A and B were found to reduce levels of NKRF and CLDN1 messenger RNA, and alter levels of other messenger RNAs that regulate intestinal permeability.

Conclusions: Based on experiments in knockout mice and analyses of intestinal tissue samples from patients with IBS-D, MIR29 targets and reduces expression of CLDN1 and NKRF to increase intestinal permeability. Strategies to block MIR29 might be developed to restore intestinal permeability in patients with IBS-D.

Keywords: Gene Regulation; Intestinal Barrier Function; Mouse Model; mRNA Processing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Case-Control Studies
  • Cell Line
  • Claudin-1 / genetics
  • Claudin-1 / metabolism*
  • Colitis / chemically induced
  • Colitis / genetics
  • Colitis / metabolism*
  • Colitis / pathology
  • Colon / metabolism*
  • Colon / pathology
  • Disease Models, Animal
  • Down-Regulation
  • Gene Knockdown Techniques
  • Genetic Predisposition to Disease
  • Humans
  • Inflammatory Bowel Diseases / genetics
  • Inflammatory Bowel Diseases / metabolism*
  • Inflammatory Bowel Diseases / pathology
  • Male
  • Mice, Inbred C57BL
  • Mice, Knockout
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Permeability
  • Phenotype
  • RNA, Messenger / metabolism
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Signal Transduction
  • Trinitrobenzenesulfonic Acid

Substances

  • CLDN1 protein, human
  • Claudin-1
  • Cldn1 protein, mouse
  • MIRN29 microRNA, human
  • MIRN29 microRNA, mouse
  • MicroRNAs
  • NKRF protein, human
  • NKRF protein, mouse
  • RNA, Messenger
  • Repressor Proteins
  • Trinitrobenzenesulfonic Acid