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MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway

Purpose. Nerve growth factor efficacy was demonstrated for corneal lesions treatment, and recombinant human NGF (rhNGF) was approved for neurotrophic keratitis therapy. However, NGF-induced molecular responses in cornea are still largely unknown. We analyzed microRNAs expression in human epithelial...

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Autores principales: Compagnoni, Chiara, Zelli, Veronica, Bianchi, Andrea, Di Marco, Antinisca, Capelli, Roberta, Vecchiotti, Davide, Brandolini, Laura, Cimini, Anna Maria, Zazzeroni, Francesca, Allegretti, Marcello, Alesse, Edoardo, Tessitore, Alessandra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8998691/
https://www.ncbi.nlm.nih.gov/pubmed/35408969
http://dx.doi.org/10.3390/ijms23073597
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author Compagnoni, Chiara
Zelli, Veronica
Bianchi, Andrea
Di Marco, Antinisca
Capelli, Roberta
Vecchiotti, Davide
Brandolini, Laura
Cimini, Anna Maria
Zazzeroni, Francesca
Allegretti, Marcello
Alesse, Edoardo
Tessitore, Alessandra
author_facet Compagnoni, Chiara
Zelli, Veronica
Bianchi, Andrea
Di Marco, Antinisca
Capelli, Roberta
Vecchiotti, Davide
Brandolini, Laura
Cimini, Anna Maria
Zazzeroni, Francesca
Allegretti, Marcello
Alesse, Edoardo
Tessitore, Alessandra
author_sort Compagnoni, Chiara
collection PubMed
description Purpose. Nerve growth factor efficacy was demonstrated for corneal lesions treatment, and recombinant human NGF (rhNGF) was approved for neurotrophic keratitis therapy. However, NGF-induced molecular responses in cornea are still largely unknown. We analyzed microRNAs expression in human epithelial corneal cells after time-dependent rhNGF treatment. Methods. Nearly 700 microRNAs were analyzed by qRT-PCR. MicroRNAs showing significant expression differences were examined by DIANA-miRpath v.3.0 to identify target genes and pathways. Immunoblots were performed to preliminarily assess the strength of the in silico results. Results. Twenty-one microRNAs (miR-26a-1-3p, miR-30d-3p, miR-27b-5p, miR-146a-5p, miR-362-5p, mir-550a-5p, mir-34a-3p, mir-1227-3p, mir-27a-5p, mir-222-5p, mir-151a-5p, miR-449a, let7c-5p, miR-337-5p, mir-29b-3p, miR-200b-3p, miR-141-3p, miR-671-3p, miR-324-5p, mir-411-3p, and mir-425-3p) were significantly regulated in response to rhNGF. In silico analysis evidenced interesting target genes and pathways, including that of neurotrophin, when analyzed in depth. Almost 80 unique target genes (e.g., PI3K, AKT, MAPK, KRAS, BRAF, RhoA, Cdc42, Rac1, Bax, Bcl2, FasL) were identified as being among those most involved in neurotrophin signaling and in controlling cell proliferation, growth, and apoptosis. AKT and RhoA immunoblots demonstrated congruence with microRNA expression, providing preliminary validation of in silico data. Conclusions. MicroRNA levels in response to rhNGF were for the first time analyzed in corneal cells. Novel insights about microRNAs, target genes, pathways modulation, and possible biological responses were provided. Importantly, given the putative role of microRNAs as biomarkers or therapeutic targets, our results make available data which might be potentially exploitable for clinical applications.
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spelling pubmed-89986912022-04-12 MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway Compagnoni, Chiara Zelli, Veronica Bianchi, Andrea Di Marco, Antinisca Capelli, Roberta Vecchiotti, Davide Brandolini, Laura Cimini, Anna Maria Zazzeroni, Francesca Allegretti, Marcello Alesse, Edoardo Tessitore, Alessandra Int J Mol Sci Article Purpose. Nerve growth factor efficacy was demonstrated for corneal lesions treatment, and recombinant human NGF (rhNGF) was approved for neurotrophic keratitis therapy. However, NGF-induced molecular responses in cornea are still largely unknown. We analyzed microRNAs expression in human epithelial corneal cells after time-dependent rhNGF treatment. Methods. Nearly 700 microRNAs were analyzed by qRT-PCR. MicroRNAs showing significant expression differences were examined by DIANA-miRpath v.3.0 to identify target genes and pathways. Immunoblots were performed to preliminarily assess the strength of the in silico results. Results. Twenty-one microRNAs (miR-26a-1-3p, miR-30d-3p, miR-27b-5p, miR-146a-5p, miR-362-5p, mir-550a-5p, mir-34a-3p, mir-1227-3p, mir-27a-5p, mir-222-5p, mir-151a-5p, miR-449a, let7c-5p, miR-337-5p, mir-29b-3p, miR-200b-3p, miR-141-3p, miR-671-3p, miR-324-5p, mir-411-3p, and mir-425-3p) were significantly regulated in response to rhNGF. In silico analysis evidenced interesting target genes and pathways, including that of neurotrophin, when analyzed in depth. Almost 80 unique target genes (e.g., PI3K, AKT, MAPK, KRAS, BRAF, RhoA, Cdc42, Rac1, Bax, Bcl2, FasL) were identified as being among those most involved in neurotrophin signaling and in controlling cell proliferation, growth, and apoptosis. AKT and RhoA immunoblots demonstrated congruence with microRNA expression, providing preliminary validation of in silico data. Conclusions. MicroRNA levels in response to rhNGF were for the first time analyzed in corneal cells. Novel insights about microRNAs, target genes, pathways modulation, and possible biological responses were provided. Importantly, given the putative role of microRNAs as biomarkers or therapeutic targets, our results make available data which might be potentially exploitable for clinical applications. MDPI 2022-03-25 /pmc/articles/PMC8998691/ /pubmed/35408969 http://dx.doi.org/10.3390/ijms23073597 Text en © 2022 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
Compagnoni, Chiara
Zelli, Veronica
Bianchi, Andrea
Di Marco, Antinisca
Capelli, Roberta
Vecchiotti, Davide
Brandolini, Laura
Cimini, Anna Maria
Zazzeroni, Francesca
Allegretti, Marcello
Alesse, Edoardo
Tessitore, Alessandra
MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway
title MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway
title_full MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway
title_fullStr MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway
title_full_unstemmed MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway
title_short MicroRNAs Expression in Response to rhNGF in Epithelial Corneal Cells: Focus on Neurotrophin Signaling Pathway
title_sort micrornas expression in response to rhngf in epithelial corneal cells: focus on neurotrophin signaling pathway
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8998691/
https://www.ncbi.nlm.nih.gov/pubmed/35408969
http://dx.doi.org/10.3390/ijms23073597
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