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

doi:10.3389/fimmu.2019.02449

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^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

30 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 language	English
Article number	2449
Journal	Frontiers in Immunology
Volume	10
DOIs	https://doi.org/10.3389/fimmu.2019.02449
State	Published - 20 Nov 2019
Externally published	Yes

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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10.3389/fimmu.2019.02449

Cite this

Dubois-Camacho, K., Diaz-Jimenez, D., De la Fuente, M., Quera, R., Simian, D., Martínez, M., Landskron, G., Olivares-Morales, M., Cidlowski, J. A., Xu, X., Gao, G., Xie, J., Chnaiderman, J., Soto-Rifo, R., González, M. J., Calixto, A., & Hermoso, M. A. (2019). Inhibition of miR-378a-3p by Inflammation Enhances IL-33 Levels: A Novel Mechanism of Alarmin Modulation in Ulcerative Colitis. Frontiers in Immunology, 10, Article 2449. https://doi.org/10.3389/fimmu.2019.02449

@article{15b19aa89cf44169a73af069e7706c92,

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

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.",

keywords = "IBD, IL-33, PPARGC1B, alarmin, miR-378a-3p, microRNA, microRNA family, ulcerative colitis",

author = "Karen Dubois-Camacho and David Diaz-Jimenez and {De la Fuente}, Marjorie and Rodrigo Quera and Daniela Simian and Maripaz Mart{\'i}nez and Glauben Landskron and Mauricio Olivares-Morales and Cidlowski, {John A.} and Xiaojiang Xu and Guangping Gao and Jun Xie and Jon{\'a}s Chnaiderman and Ricardo Soto-Rifo and Gonz{\'a}lez, {Mar{\'i}a Julieta} and Andrea Calixto and Hermoso, {Marcela A.}",

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: Cl{\'i}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). Publisher Copyright: {\textcopyright} Copyright {\textcopyright} 2019 Dubois-Camacho, Diaz-Jimenez, De la Fuente, Quera, Simian, Mart{\'i}nez, Landskron, Olivares-Morales, Cidlowski, Xu, Gao, Xie, Chnaiderman, Soto-Rifo, Gonz{\'a}lez, Calixto and Hermoso.",

year = "2019",

month = nov,

day = "20",

doi = "10.3389/fimmu.2019.02449",

language = "English",

volume = "10",

journal = "Frontiers in Immunology",

issn = "1664-3224",

publisher = "Frontiers Media S.A.",

}

Dubois-Camacho, K, Diaz-Jimenez, D, De la Fuente, M, Quera, R, Simian, D, Martínez, M, Landskron, G, Olivares-Morales, M, Cidlowski, JA, Xu, X, Gao, G, Xie, J, Chnaiderman, J, Soto-Rifo, R, González, MJ, Calixto, A & Hermoso, MA 2019, 'Inhibition of miR-378a-3p by Inflammation Enhances IL-33 Levels: A Novel Mechanism of Alarmin Modulation in Ulcerative Colitis', Frontiers in Immunology, vol. 10, 2449. https://doi.org/10.3389/fimmu.2019.02449

TY - JOUR

T1 - Inhibition of miR-378a-3p by Inflammation Enhances IL-33 Levels

T2 - A Novel Mechanism of Alarmin Modulation in Ulcerative Colitis

AU - Dubois-Camacho, Karen

AU - Diaz-Jimenez, David

AU - De la Fuente, Marjorie

AU - Quera, Rodrigo

AU - Simian, Daniela

AU - Martínez, Maripaz

AU - Landskron, Glauben

AU - Olivares-Morales, Mauricio

AU - Cidlowski, John A.

AU - Xu, Xiaojiang

AU - Gao, Guangping

AU - Xie, Jun

AU - Chnaiderman, Jonás

AU - Soto-Rifo, Ricardo

AU - González, María Julieta

AU - Calixto, Andrea

AU - Hermoso, Marcela A.

N1 - 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.

PY - 2019/11/20

Y1 - 2019/11/20

N2 - 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.

AB - 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.

KW - IBD

KW - IL-33

KW - PPARGC1B

KW - alarmin

KW - miR-378a-3p

KW - microRNA

KW - microRNA family

KW - ulcerative colitis

UR - http://www.scopus.com/inward/record.url?scp=85076346182&partnerID=8YFLogxK

U2 - 10.3389/fimmu.2019.02449

DO - 10.3389/fimmu.2019.02449

M3 - Article

C2 - 31824476

AN - SCOPUS:85076346182

SN - 1664-3224

VL - 10

JO - Frontiers in Immunology

JF - Frontiers in Immunology

M1 - 2449

ER -

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

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Keywords

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