The University of Southampton
University of Southampton Institutional Repository

MicroRNA23a overexpression in Crohn’s Disease targets Tumour Necrosis Factor Alpha Inhibitor Protein 3, increasing sensitivity to TNF and modifying the epithelial barrier

MicroRNA23a overexpression in Crohn’s Disease targets Tumour Necrosis Factor Alpha Inhibitor Protein 3, increasing sensitivity to TNF and modifying the epithelial barrier
MicroRNA23a overexpression in Crohn’s Disease targets Tumour Necrosis Factor Alpha Inhibitor Protein 3, increasing sensitivity to TNF and modifying the epithelial barrier
Background and Aims: Mucosal healing is important in Crohn’s disease therapies. Epithelial homeostasis becomes dysregulated in Crohn’s with increased permeability, inflammation and diarrhoea. MicroRNAs are small non-coding RNAs, which regulate gene expression and show changes in inflammatory bowel disease. Tumour Necrosis Factor Alpha Inhibitor Protein 3 is raised in Crohn’s and regulates TNFα-mediated activation of NFκB. We investigated TNFα regulation by microRNA in Crohn’s disease and studied effects on epithelial permeability and inflammation.

Methods: Colonic epithelium from CD and healthy donor biopsies was isolated using laser capture microdissection and microRNA quantified. Tumour Necrosis Factor Alpha Inhibitor Protein 3 was characterized immunohistochemically on serial sections. Expression effect of microRNA was confirmed with luciferase reporter assays. Functional barrier permeability studies and innate cytokine release were investigated with cell and explant culture studies.

Results: MicroRNA23a levels significantly increased in colonic Crohn’s epithelium compared to healthy. Luciferase reporter assays in transfected epithelial cells confirmed that microRNA23a repressed expression via the 3’ untranslated region of Tumour Necrosis Factor Alpha Inhibitor Protein 3 mRNA coinciding with increased NFκB-mediated transcription. Immunohistochemical staining of TNFAIP3 protein in colonic biopsies was reduced or absent in adjacent Crohn’s sections, correlating inversely with microRNA23a levels and encompassing some inter-cohort variation. Overexpression of microRNA23a increased epithelial barrier permeability in a colonic epithelial model and increased inflammatory cytokine release in cultured explant biopsies, mimicking Crohn’s disease characteristics.

Conclusion: MicroRNA23a overexpression in colonic Crohn’s epithelium represses Tumour Necrosis Factor Alpha Inhibitor Protein 3, enhancing sensitivity to TNFα with increased intestinal permeability and cytokine release.
microRNA, epithelium, TNFAIP3, inflammatory bowel disease, Crohn's disease, TNF
1873-9946
1-12
Felwick, Richard
b19f1ab6-1de4-4df6-b149-4bc337c3fcb9
Dingley, Geraint
f955289e-1a55-4d45-b65e-446d67f48a7d
Martinez-Nunez, Rocio T.
3b2f9516-1067-4c16-90f9-e13dad821666
Sanchez-Elsner, Tilman
b8799f8d-e2b4-4b37-b77c-f2f0e8e2070d
Cummings, J.R. Fraser
89e8e80c-b6e8-4387-a63c-3796b5ad7e14
Collins, Jane
be0e66f1-3036-47fa-9d7e-914c48710ba4
Felwick, Richard
b19f1ab6-1de4-4df6-b149-4bc337c3fcb9
Dingley, Geraint
f955289e-1a55-4d45-b65e-446d67f48a7d
Martinez-Nunez, Rocio T.
3b2f9516-1067-4c16-90f9-e13dad821666
Sanchez-Elsner, Tilman
b8799f8d-e2b4-4b37-b77c-f2f0e8e2070d
Cummings, J.R. Fraser
89e8e80c-b6e8-4387-a63c-3796b5ad7e14
Collins, Jane
be0e66f1-3036-47fa-9d7e-914c48710ba4

Felwick, Richard, Dingley, Geraint, Martinez-Nunez, Rocio T., Sanchez-Elsner, Tilman, Cummings, J.R. Fraser and Collins, Jane (2019) MicroRNA23a overexpression in Crohn’s Disease targets Tumour Necrosis Factor Alpha Inhibitor Protein 3, increasing sensitivity to TNF and modifying the epithelial barrier. Journal of Crohn's and Colitis, 1-12. (doi:10.1093/ecco-jcc/jjz145). (In Press)

Record type: Article

Abstract

Background and Aims: Mucosal healing is important in Crohn’s disease therapies. Epithelial homeostasis becomes dysregulated in Crohn’s with increased permeability, inflammation and diarrhoea. MicroRNAs are small non-coding RNAs, which regulate gene expression and show changes in inflammatory bowel disease. Tumour Necrosis Factor Alpha Inhibitor Protein 3 is raised in Crohn’s and regulates TNFα-mediated activation of NFκB. We investigated TNFα regulation by microRNA in Crohn’s disease and studied effects on epithelial permeability and inflammation.

Methods: Colonic epithelium from CD and healthy donor biopsies was isolated using laser capture microdissection and microRNA quantified. Tumour Necrosis Factor Alpha Inhibitor Protein 3 was characterized immunohistochemically on serial sections. Expression effect of microRNA was confirmed with luciferase reporter assays. Functional barrier permeability studies and innate cytokine release were investigated with cell and explant culture studies.

Results: MicroRNA23a levels significantly increased in colonic Crohn’s epithelium compared to healthy. Luciferase reporter assays in transfected epithelial cells confirmed that microRNA23a repressed expression via the 3’ untranslated region of Tumour Necrosis Factor Alpha Inhibitor Protein 3 mRNA coinciding with increased NFκB-mediated transcription. Immunohistochemical staining of TNFAIP3 protein in colonic biopsies was reduced or absent in adjacent Crohn’s sections, correlating inversely with microRNA23a levels and encompassing some inter-cohort variation. Overexpression of microRNA23a increased epithelial barrier permeability in a colonic epithelial model and increased inflammatory cytokine release in cultured explant biopsies, mimicking Crohn’s disease characteristics.

Conclusion: MicroRNA23a overexpression in colonic Crohn’s epithelium represses Tumour Necrosis Factor Alpha Inhibitor Protein 3, enhancing sensitivity to TNFα with increased intestinal permeability and cytokine release.

Text
Felwick R2 Manuscript Full figs Revised - Accepted Manuscript
Restricted to Repository staff only until 9 August 2020.
Request a copy
Text
Felwick et al jjz145-1corr - Proof
Restricted to Repository staff only
Request a copy
Text
Felwick R2 Supplementary Information
Restricted to Repository staff only until 9 August 2020.
Request a copy

More information

In preparation date: 2018
Submitted date: 15 December 2018
Accepted/In Press date: 9 August 2019
Keywords: microRNA, epithelium, TNFAIP3, inflammatory bowel disease, Crohn's disease, TNF

Identifiers

Local EPrints ID: 435027
URI: https://eprints.soton.ac.uk/id/eprint/435027
ISSN: 1873-9946
PURE UUID: e39cc2f3-2682-482d-a659-c5108024fcd6

Catalogue record

Date deposited: 18 Oct 2019 16:30
Last modified: 18 Oct 2019 16:30

Export record

Altmetrics

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×