Inhibition of miR-378a-3p by Inflammation Enhances IL-33 Levels: A Novel Mechanism of Alarmin Modulation in Ulcerative Colitis

Karen Dubois-Camacho, David Diaz-Jimenez, Marjorie De la Fuente, Rodrigo Quera, Daniela Simian, Maripaz Martínez, Glauben Landskron, Mauricio Olivares-Morales, John A. Cidlowski, Xiaojiang Xu, Guangping Gao, Jun Xie, Jonás Chnaiderman, Ricardo Soto-Rifo, María Julieta González, Andrea Calixto, Marcela A. Hermoso

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9 Scopus citations

Abstract

Ulcerative colitis (UC) is an inflammatory bowel disease (IBD) characterized by mucosa damage associated with an uncontrolled inflammatory response. This immunological impairment leads to altered inflammatory mediators such as IL-33, which is shown to increase in the mucosa of active UC (aUC) patients. MicroRNAs present a distorted feature in inflamed colonic mucosa and are potential IL-33 regulating candidates in UC. Therefore, we studied the microRNA and mRNA profiles in inflamed colonic samples of UC patients, evaluating the effect of a microRNA (selected by in silico analysis and its expression in UC patients), on IL-33 under inflammatory conditions. We found that inflamed mucosa (n = 8) showed increased expression of 40 microRNAs and 2,120 mRNAs, while 49 microRNAs and 1,734 mRNAs were decreased, as determined by microarrays. In particular, IL-33 mRNA showed a 3.8-fold increase and eight members of a microRNA family (miR-378), which targets IL-33 mRNA in the 3′UTR, were decreased (−3.9 to −3.0 times). We selected three members of the miR-378 family (miR-378a-3p, miR-422a, and miR-378c) according to background information and interaction energy analysis, for further correlation analyses with IL-33 expression through qPCR and ELISA, respectively. We determined that aUC (n = 24) showed high IL-33 levels, and decreased expression of miR-378a-3p and miR-422a compared to inactive UC (n = 10) and controls (n = 6). Moreover, both microRNAs were inversely correlated with IL-33 expression, while miR-378c does not show a significant difference. To evaluate the effect of TNFα on the studied microRNAs, aUC patients with anti-TNF therapy were compared to aUC receiving other treatments. The levels of miR-378a-3p and miR-378c were higher in aUC patients with anti-TNF. Based on these findings, we selected miR-378a-3p to exploring the molecular mechanism involved by in vitro assays, showing that over-expression of miR-378a-3p decreased the levels of an IL-33 target sequence β-gal-reporter gene in HEK293 cells. Stable miR-378a-3p over-expression/inhibition inversely modulated IL-33 content and altered viability of HT-29 cells. Additionally, in an inflammatory context, TNFα decreased miR-378a-3p levels in HT-29 cells enhancing IL-33 expression. Together, our results propose a regulatory mechanism of IL-33 expression exerted by miR-378a-3p in an inflammatory environment, contributing to the understanding of UC pathogenesis.

Original languageEnglish
Article number2449
JournalFrontiers in Immunology
Volume10
DOIs
StatePublished - 20 Nov 2019
Externally publishedYes

Bibliographical note

Funding Information:
We wish to thank Gene Therapy Center UMASS (MA, USA); the NIEHS transduction laboratory (NC, USA); the Center for Genomics and Bioinformatics, Universidad Mayor (Santiago, Chile); Cell Transformation Laboratory and Cancer Genomic Laboratory, Universidad de Chile (Santiago, Chile) and Laboratory of Immunogastroenterology, Hospital Cl?nico Universidad de Chile (Santiago, Chile) for their support to this research. The figures were produced using Servier Medical Art from www.servier.com. We would also like to thank David Cox for his editing contribution. Funding. Cl?nica Las Condes Academic Project PI2013-B002 (RQ), National Fund for Scientific and Technological Development No. 1170648 (MH); National Commission for Scientific and Technological Research Scholarship No. 21150264 (DD-J), No. 21120682 (MO-M); and MECESUP Scholarship No. UCH 0714 (KD-C).

Funding Information:
Clínica Las Condes Academic Project PI2013-B002 (RQ), National Fund for Scientific and Technological Development No. 1170648 (MH); National Commission for Scientific and Technological Research Scholarship No. 21150264 (DD-J), No. 21120682 (MO-M); and MECESUP Scholarship No. UCH 0714 (KD-C).

Publisher Copyright:
© Copyright © 2019 Dubois-Camacho, Diaz-Jimenez, De la Fuente, Quera, Simian, Martínez, Landskron, Olivares-Morales, Cidlowski, Xu, Gao, Xie, Chnaiderman, Soto-Rifo, González, Calixto and Hermoso.

Keywords

  • IBD
  • IL-33
  • PPARGC1B
  • alarmin
  • miR-378a-3p
  • microRNA
  • microRNA family
  • ulcerative colitis

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