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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 that regulate gene expression and show changes in inflammatory bowel disease. Tumour necrosis factor alpha [TNFα] 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 [CD], 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 was quantified. Tumour necrosis factor alpha inhibitor protein 3 was characterised 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 with healthy epithelium. 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 intercohort 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. Conclusions: 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.

Crohn's, MicroRNA, TNF, TNFAIP3, epithelium, inflammatory bowel disease
1873-9946
381-392
Felwick, Richard
b19f1ab6-1de4-4df6-b149-4bc337c3fcb9
Dingley, Geraint
f955289e-1a55-4d45-b65e-446d67f48a7d
Martinez-Nunez, Rocio T.
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Sanchez-Elsner, Tilman
b8799f8d-e2b4-4b37-b77c-f2f0e8e2070d
Cummings, J.R. Fraser
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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 (2020) 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, 14 (3), 381-392. (doi:10.1093/ecco-jcc/jjz145).

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 that regulate gene expression and show changes in inflammatory bowel disease. Tumour necrosis factor alpha [TNFα] 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 [CD], 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 was quantified. Tumour necrosis factor alpha inhibitor protein 3 was characterised 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 with healthy epithelium. 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 intercohort 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. Conclusions: 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.

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More information

In preparation date: 2018
Submitted date: 15 December 2018
Accepted/In Press date: 9 August 2019
e-pub ahead of print date: 18 October 2019
Published date: 13 March 2020
Keywords: Crohn's, MicroRNA, TNF, TNFAIP3, epithelium, inflammatory bowel disease

Identifiers

Local EPrints ID: 435027
URI: http://eprints.soton.ac.uk/id/eprint/435027
ISSN: 1873-9946
PURE UUID: e39cc2f3-2682-482d-a659-c5108024fcd6
ORCID for Tilman Sanchez-Elsner: ORCID iD orcid.org/0000-0003-1915-2410

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Date deposited: 18 Oct 2019 16:30
Last modified: 26 Nov 2021 06:32

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Contributors

Author: Richard Felwick
Author: Geraint Dingley
Author: Rocio T. Martinez-Nunez
Author: J.R. Fraser Cummings
Author: Jane Collins

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