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miR-199a-5p inhibits the expression of ABCB11 in obstructive cholestasis

ATP-binding cassette, subfamily B member 11 (ABCB11) is an efflux transporter for bile acids on the liver canalicular membrane. The expression of this transporter is reduced in cholestasis; however, the mechanisms contributing to this reduction are unclear. In this study, we sought to determine whet...

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Autores principales: Balasubramaniyan, Natarajan, Devereaux, Michael W., Orlicky, David J., Sokol, Ronald J., Suchy, Frederick J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8665360/
https://www.ncbi.nlm.nih.gov/pubmed/34774795
http://dx.doi.org/10.1016/j.jbc.2021.101400
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author Balasubramaniyan, Natarajan
Devereaux, Michael W.
Orlicky, David J.
Sokol, Ronald J.
Suchy, Frederick J.
author_facet Balasubramaniyan, Natarajan
Devereaux, Michael W.
Orlicky, David J.
Sokol, Ronald J.
Suchy, Frederick J.
author_sort Balasubramaniyan, Natarajan
collection PubMed
description ATP-binding cassette, subfamily B member 11 (ABCB11) is an efflux transporter for bile acids on the liver canalicular membrane. The expression of this transporter is reduced in cholestasis; however, the mechanisms contributing to this reduction are unclear. In this study, we sought to determine whether miR-199a-5p contributes to the depletion of ABCB11/Abcb11 in cholestasis in mice. In a microRNA (miRNA) screen of mouse liver after common bile duct ligation (CBDL), we found that miR-199a-5p was significantly upregulated by approximately fourfold. In silico analysis predicted that miR-199a-5p would target the 3′-untranslated region (3′-UTR) of ABCB11/Abcb11 mRNA. The expression of ABCB11-3′-UTR luciferase construct in Huh-7 cells was markedly inhibited by cotransfection of a miRNA-199a-5p mimic, which was reversed by an miRNA-199a-5p mimic inhibitor. We also show treatment of mice after CBDL with the potent nuclear receptor FXR agonist obeticholic acid (OCA) significantly increased Abcb11 mRNA and protein and decreased miR-199a-5p expression. Computational mapping revealed a well-conserved FXR-binding site (FXRE) in the promoter of the gene encoding miR-199a-5, termed miR199a-2. Electromobility shift, chromatin immunoprecipitation, and miR199a-2 promoter-luciferase assays confirmed that this binding site was functional. Finally, CBDL in mice led to depletion of nuclear repressor NcoR1 binding at the miR199a-2 promoter, which facilitates transcription of miR199a-2. In CBDL mice treated with OCA, NcoR1 recruitment to the miR199a-2 FXRE was maintained at levels found in sham-operated mice. In conclusion, we demonstrate that miR-199a-5p is involved in regulating ABCB11/Abcb11 expression, is aberrantly upregulated in obstructive cholestasis, and is downregulated by the FXR agonist OCA.
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spelling pubmed-86653602021-12-21 miR-199a-5p inhibits the expression of ABCB11 in obstructive cholestasis Balasubramaniyan, Natarajan Devereaux, Michael W. Orlicky, David J. Sokol, Ronald J. Suchy, Frederick J. J Biol Chem Research Article ATP-binding cassette, subfamily B member 11 (ABCB11) is an efflux transporter for bile acids on the liver canalicular membrane. The expression of this transporter is reduced in cholestasis; however, the mechanisms contributing to this reduction are unclear. In this study, we sought to determine whether miR-199a-5p contributes to the depletion of ABCB11/Abcb11 in cholestasis in mice. In a microRNA (miRNA) screen of mouse liver after common bile duct ligation (CBDL), we found that miR-199a-5p was significantly upregulated by approximately fourfold. In silico analysis predicted that miR-199a-5p would target the 3′-untranslated region (3′-UTR) of ABCB11/Abcb11 mRNA. The expression of ABCB11-3′-UTR luciferase construct in Huh-7 cells was markedly inhibited by cotransfection of a miRNA-199a-5p mimic, which was reversed by an miRNA-199a-5p mimic inhibitor. We also show treatment of mice after CBDL with the potent nuclear receptor FXR agonist obeticholic acid (OCA) significantly increased Abcb11 mRNA and protein and decreased miR-199a-5p expression. Computational mapping revealed a well-conserved FXR-binding site (FXRE) in the promoter of the gene encoding miR-199a-5, termed miR199a-2. Electromobility shift, chromatin immunoprecipitation, and miR199a-2 promoter-luciferase assays confirmed that this binding site was functional. Finally, CBDL in mice led to depletion of nuclear repressor NcoR1 binding at the miR199a-2 promoter, which facilitates transcription of miR199a-2. In CBDL mice treated with OCA, NcoR1 recruitment to the miR199a-2 FXRE was maintained at levels found in sham-operated mice. In conclusion, we demonstrate that miR-199a-5p is involved in regulating ABCB11/Abcb11 expression, is aberrantly upregulated in obstructive cholestasis, and is downregulated by the FXR agonist OCA. American Society for Biochemistry and Molecular Biology 2021-11-12 /pmc/articles/PMC8665360/ /pubmed/34774795 http://dx.doi.org/10.1016/j.jbc.2021.101400 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Balasubramaniyan, Natarajan
Devereaux, Michael W.
Orlicky, David J.
Sokol, Ronald J.
Suchy, Frederick J.
miR-199a-5p inhibits the expression of ABCB11 in obstructive cholestasis
title miR-199a-5p inhibits the expression of ABCB11 in obstructive cholestasis
title_full miR-199a-5p inhibits the expression of ABCB11 in obstructive cholestasis
title_fullStr miR-199a-5p inhibits the expression of ABCB11 in obstructive cholestasis
title_full_unstemmed miR-199a-5p inhibits the expression of ABCB11 in obstructive cholestasis
title_short miR-199a-5p inhibits the expression of ABCB11 in obstructive cholestasis
title_sort mir-199a-5p inhibits the expression of abcb11 in obstructive cholestasis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8665360/
https://www.ncbi.nlm.nih.gov/pubmed/34774795
http://dx.doi.org/10.1016/j.jbc.2021.101400
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