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Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells

We previously reported anti-miR-328 therapy for dry eye disease (DED). Since decreased mucin secretion is a risk factor for DED, we aimed to explore whether anti-miR-328 affects mucin expression and goblet cells. MiR-328 was increased in goblet cells when they were under desiccating stress or treate...

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Autores principales: Choo, Jackson, Liao, Chun-Huei, Tseng, Ching-Li, Chen, Jiunn-Liang, Cheng, Huey-Chuan, Liang, Chung-Ling, Juo, Suh-Hang Hank
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953597/
https://www.ncbi.nlm.nih.gov/pubmed/36830824
http://dx.doi.org/10.3390/biomedicines11020287
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author Choo, Jackson
Liao, Chun-Huei
Tseng, Ching-Li
Chen, Jiunn-Liang
Cheng, Huey-Chuan
Liang, Chung-Ling
Juo, Suh-Hang Hank
author_facet Choo, Jackson
Liao, Chun-Huei
Tseng, Ching-Li
Chen, Jiunn-Liang
Cheng, Huey-Chuan
Liang, Chung-Ling
Juo, Suh-Hang Hank
author_sort Choo, Jackson
collection PubMed
description We previously reported anti-miR-328 therapy for dry eye disease (DED). Since decreased mucin secretion is a risk factor for DED, we aimed to explore whether anti-miR-328 affects mucin expression and goblet cells. MiR-328 was increased in goblet cells when they were under desiccating stress or treated with benzalkonium chloride (BAC), both of which are risk factors for DED. Based on bioinformatics tool results, miR-328 was predicted to directly target the transcription factor CREB1 that has been known to promote the expression of mucin5AC. The inhibitory effect of miR-328 on CREB1 was confirmed by the transfection assay. A miR-328 binding site on the CREB1 gene was confirmed by the luciferase assay. Furthermore, anti-miR-328 increased CREB1 and mucin5AC in cultured goblet cells according to qPCR, Western blot, and IF staining experiments. Anti-miR-328 increased mucin5AC secretion from the cultured goblet cells based on an ELISA assay for the cultured medium. Finally, impression cytology data revealed anti-miR-328 increased conjunctival goblet cells in the DED rabbits induced by BAC. In conclusion, anti-miR-328 increases CREB1 expression leading to an increase in mucin5AC production and secretion. Furthermore, anti-miR-328 also increases conjunctival goblet cells. These results warrant the further development of anti-miR-328 therapy for DED.
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spelling pubmed-99535972023-02-25 Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells Choo, Jackson Liao, Chun-Huei Tseng, Ching-Li Chen, Jiunn-Liang Cheng, Huey-Chuan Liang, Chung-Ling Juo, Suh-Hang Hank Biomedicines Article We previously reported anti-miR-328 therapy for dry eye disease (DED). Since decreased mucin secretion is a risk factor for DED, we aimed to explore whether anti-miR-328 affects mucin expression and goblet cells. MiR-328 was increased in goblet cells when they were under desiccating stress or treated with benzalkonium chloride (BAC), both of which are risk factors for DED. Based on bioinformatics tool results, miR-328 was predicted to directly target the transcription factor CREB1 that has been known to promote the expression of mucin5AC. The inhibitory effect of miR-328 on CREB1 was confirmed by the transfection assay. A miR-328 binding site on the CREB1 gene was confirmed by the luciferase assay. Furthermore, anti-miR-328 increased CREB1 and mucin5AC in cultured goblet cells according to qPCR, Western blot, and IF staining experiments. Anti-miR-328 increased mucin5AC secretion from the cultured goblet cells based on an ELISA assay for the cultured medium. Finally, impression cytology data revealed anti-miR-328 increased conjunctival goblet cells in the DED rabbits induced by BAC. In conclusion, anti-miR-328 increases CREB1 expression leading to an increase in mucin5AC production and secretion. Furthermore, anti-miR-328 also increases conjunctival goblet cells. These results warrant the further development of anti-miR-328 therapy for DED. MDPI 2023-01-19 /pmc/articles/PMC9953597/ /pubmed/36830824 http://dx.doi.org/10.3390/biomedicines11020287 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Choo, Jackson
Liao, Chun-Huei
Tseng, Ching-Li
Chen, Jiunn-Liang
Cheng, Huey-Chuan
Liang, Chung-Ling
Juo, Suh-Hang Hank
Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells
title Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells
title_full Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells
title_fullStr Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells
title_full_unstemmed Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells
title_short Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells
title_sort inhibition of microrna-328 increases ocular mucin expression and conjunctival goblet cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953597/
https://www.ncbi.nlm.nih.gov/pubmed/36830824
http://dx.doi.org/10.3390/biomedicines11020287
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