Cargando…

The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes

Corneal transparency, necessary for vision and depending on the high organization of stromal extracellular matrix, is maintained by keratocytes. Severe or continuous corneal injuries determine exaggerated healing responses resulting in the formation of irreversible fibrotic scars and vision impairme...

Descripción completa

Detalles Bibliográficos
Autores principales: Rosa, Irene, Fioretto, Bianca Saveria, Romano, Eloisa, Buzzi, Matilde, Mencucci, Rita, Marini, Mirca, Manetti, Mirko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9737374/
https://www.ncbi.nlm.nih.gov/pubmed/36499651
http://dx.doi.org/10.3390/ijms232315325
_version_ 1784847270313721856
author Rosa, Irene
Fioretto, Bianca Saveria
Romano, Eloisa
Buzzi, Matilde
Mencucci, Rita
Marini, Mirca
Manetti, Mirko
author_facet Rosa, Irene
Fioretto, Bianca Saveria
Romano, Eloisa
Buzzi, Matilde
Mencucci, Rita
Marini, Mirca
Manetti, Mirko
author_sort Rosa, Irene
collection PubMed
description Corneal transparency, necessary for vision and depending on the high organization of stromal extracellular matrix, is maintained by keratocytes. Severe or continuous corneal injuries determine exaggerated healing responses resulting in the formation of irreversible fibrotic scars and vision impairment. Soluble guanylate cyclase (sGC) stimulation demonstrated antifibrotic effects in both experimental fibrosis and human lung and skin fibroblasts. Here, we assessed whether sGC stimulation with BAY 41-2272 could attenuate transforming growth factor β1 (TGFβ1)-induced myofibroblast differentiation of human corneal keratocytes. Cells were challenged with TGFβ1, with/without BAY 41-2272 preincubation, and subsequently assessed for viability, proliferation, migration, chemoinvasion, as well for the expression of myofibroblast/fibroblast activation markers and contractile abilities. Treatment with BAY 41-2272 did not affect keratocyte viability, while preincubation of cells with the sGC stimulator was able to inhibit TGFβ1-induced proliferation, wound healing capacity, and invasiveness. BAY 41-2272 was also able to attenuate TGFβ1-induced myofibroblast-like profibrotic phenotype of keratocytes, as demonstrated by the significant decrease in ACTA2, COL1A1, COL1A2, FN1 and PDPN gene expression, as well as in α-smooth muscle actin, α-1 chain of type I collagen, podoplanin, vimentin and N-cadherin protein expression. Finally, BAY 41-2272 significantly counteracted the TGFβ1-induced myofibroblast-like ability of keratocytes to contract collagen gels, reduced phosphorylated Smad3 protein levels, and attenuated gene expression of proinflammatory cytokines. Collectively, our data show for the first time that BAY 41-2272 is effective in counteracting keratocyte-to-myofibroblast transition, thus providing the rationale for the development of sGC stimulators as novel promising modulators of corneal scarring and fibrosis.
format Online
Article
Text
id pubmed-9737374
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-97373742022-12-11 The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes Rosa, Irene Fioretto, Bianca Saveria Romano, Eloisa Buzzi, Matilde Mencucci, Rita Marini, Mirca Manetti, Mirko Int J Mol Sci Article Corneal transparency, necessary for vision and depending on the high organization of stromal extracellular matrix, is maintained by keratocytes. Severe or continuous corneal injuries determine exaggerated healing responses resulting in the formation of irreversible fibrotic scars and vision impairment. Soluble guanylate cyclase (sGC) stimulation demonstrated antifibrotic effects in both experimental fibrosis and human lung and skin fibroblasts. Here, we assessed whether sGC stimulation with BAY 41-2272 could attenuate transforming growth factor β1 (TGFβ1)-induced myofibroblast differentiation of human corneal keratocytes. Cells were challenged with TGFβ1, with/without BAY 41-2272 preincubation, and subsequently assessed for viability, proliferation, migration, chemoinvasion, as well for the expression of myofibroblast/fibroblast activation markers and contractile abilities. Treatment with BAY 41-2272 did not affect keratocyte viability, while preincubation of cells with the sGC stimulator was able to inhibit TGFβ1-induced proliferation, wound healing capacity, and invasiveness. BAY 41-2272 was also able to attenuate TGFβ1-induced myofibroblast-like profibrotic phenotype of keratocytes, as demonstrated by the significant decrease in ACTA2, COL1A1, COL1A2, FN1 and PDPN gene expression, as well as in α-smooth muscle actin, α-1 chain of type I collagen, podoplanin, vimentin and N-cadherin protein expression. Finally, BAY 41-2272 significantly counteracted the TGFβ1-induced myofibroblast-like ability of keratocytes to contract collagen gels, reduced phosphorylated Smad3 protein levels, and attenuated gene expression of proinflammatory cytokines. Collectively, our data show for the first time that BAY 41-2272 is effective in counteracting keratocyte-to-myofibroblast transition, thus providing the rationale for the development of sGC stimulators as novel promising modulators of corneal scarring and fibrosis. MDPI 2022-12-05 /pmc/articles/PMC9737374/ /pubmed/36499651 http://dx.doi.org/10.3390/ijms232315325 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
Rosa, Irene
Fioretto, Bianca Saveria
Romano, Eloisa
Buzzi, Matilde
Mencucci, Rita
Marini, Mirca
Manetti, Mirko
The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes
title The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes
title_full The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes
title_fullStr The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes
title_full_unstemmed The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes
title_short The Soluble Guanylate Cyclase Stimulator BAY 41-2272 Attenuates Transforming Growth Factor β1-Induced Myofibroblast Differentiation of Human Corneal Keratocytes
title_sort soluble guanylate cyclase stimulator bay 41-2272 attenuates transforming growth factor β1-induced myofibroblast differentiation of human corneal keratocytes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9737374/
https://www.ncbi.nlm.nih.gov/pubmed/36499651
http://dx.doi.org/10.3390/ijms232315325
work_keys_str_mv AT rosairene thesolubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT fiorettobiancasaveria thesolubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT romanoeloisa thesolubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT buzzimatilde thesolubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT mencuccirita thesolubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT marinimirca thesolubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT manettimirko thesolubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT rosairene solubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT fiorettobiancasaveria solubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT romanoeloisa solubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT buzzimatilde solubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT mencuccirita solubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT marinimirca solubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes
AT manettimirko solubleguanylatecyclasestimulatorbay412272attenuatestransforminggrowthfactorb1inducedmyofibroblastdifferentiationofhumancornealkeratocytes