Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast Differentiation
Transforming Growth Factor-Beta 1 (TGF-β1) plays a pivotal role in the differentiation of fibroblasts into myofibroblasts, which is a critical process in tissue repair, fibrosis, and wound healing. Upon exposure to TGF-β1, fibroblasts acquire a contractile phenotype and secrete collagen and extracel...
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MDPI AG
2025-04-01
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| Series: | Antioxidants |
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| author | Marzieh Aminzadehanboohi Manousos Makridakis Delphine Rasti Yves Cambet Karl-Heinz Krause Antonia Vlahou Vincent Jaquet |
| author_facet | Marzieh Aminzadehanboohi Manousos Makridakis Delphine Rasti Yves Cambet Karl-Heinz Krause Antonia Vlahou Vincent Jaquet |
| author_sort | Marzieh Aminzadehanboohi |
| collection | DOAJ |
| description | Transforming Growth Factor-Beta 1 (TGF-β1) plays a pivotal role in the differentiation of fibroblasts into myofibroblasts, which is a critical process in tissue repair, fibrosis, and wound healing. Upon exposure to TGF-β1, fibroblasts acquire a contractile phenotype and secrete collagen and extracellular matrix components. Numerous studies have identified hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) as a key downstream effector of TGF-β1 in this pathway. H<sub>2</sub>O<sub>2</sub> functions as a signalling molecule, regulating various cellular processes mostly through post-translational redox modifications of cysteine thiol groups of specific proteins. In this study, we used primary human skin fibroblast cultures to investigate the oxidative mechanisms triggered by TGF-β1. We analyzed the expression of redox-related genes, evaluated the effects of the genetic and pharmacological inhibition of H<sub>2</sub>O<sub>2</sub>-producing enzymes, and employed an unbiased redox proteomics approach (OxICAT) to identify proteins undergoing reversible cysteine oxidation. Our findings revealed that TGF-β1 treatment upregulated the expression of oxidant-generating genes while downregulating antioxidant genes. Low concentrations of diphenyleneiodonium mitigated myofibroblast differentiation and mitochondrial oxygen consumption, suggesting the involvement of a flavoenzyme in this process. Furthermore, we identified the increased oxidation of highly conserved cysteine residues in key proteins such as the epidermal growth factor receptor, filamin A, fibulin-2, and endosialin during the differentiation process. Collectively, this study provides insights into the sources of H<sub>2</sub>O<sub>2</sub> in fibroblasts and highlights the novel redox mechanisms underpinning fibroblast-to-myofibroblast differentiation. |
| format | Article |
| id | doaj-art-cf12aed6dd2f4b008be91bdf1e5f764d |
| institution | DOAJ |
| issn | 2076-3921 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj-art-cf12aed6dd2f4b008be91bdf1e5f764d2025-08-20T03:14:16ZengMDPI AGAntioxidants2076-39212025-04-0114448610.3390/antiox14040486Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast DifferentiationMarzieh Aminzadehanboohi0Manousos Makridakis1Delphine Rasti2Yves Cambet3Karl-Heinz Krause4Antonia Vlahou5Vincent Jaquet6Department of Pathology and Immunology, Medical School, University of Geneva, 1211 Geneva, SwitzerlandCenter of Systems Biology, Biomedical Research Foundation Academy of Athens, 11527 Athens, GreeceDepartment of Pathology and Immunology, Medical School, University of Geneva, 1211 Geneva, SwitzerlandREADS Unit, Medical School, University of Geneva, 1211 Geneva, SwitzerlandDepartment of Pathology and Immunology, Medical School, University of Geneva, 1211 Geneva, SwitzerlandCenter of Systems Biology, Biomedical Research Foundation Academy of Athens, 11527 Athens, GreeceDepartment of Pathology and Immunology, Medical School, University of Geneva, 1211 Geneva, SwitzerlandTransforming Growth Factor-Beta 1 (TGF-β1) plays a pivotal role in the differentiation of fibroblasts into myofibroblasts, which is a critical process in tissue repair, fibrosis, and wound healing. Upon exposure to TGF-β1, fibroblasts acquire a contractile phenotype and secrete collagen and extracellular matrix components. Numerous studies have identified hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) as a key downstream effector of TGF-β1 in this pathway. H<sub>2</sub>O<sub>2</sub> functions as a signalling molecule, regulating various cellular processes mostly through post-translational redox modifications of cysteine thiol groups of specific proteins. In this study, we used primary human skin fibroblast cultures to investigate the oxidative mechanisms triggered by TGF-β1. We analyzed the expression of redox-related genes, evaluated the effects of the genetic and pharmacological inhibition of H<sub>2</sub>O<sub>2</sub>-producing enzymes, and employed an unbiased redox proteomics approach (OxICAT) to identify proteins undergoing reversible cysteine oxidation. Our findings revealed that TGF-β1 treatment upregulated the expression of oxidant-generating genes while downregulating antioxidant genes. Low concentrations of diphenyleneiodonium mitigated myofibroblast differentiation and mitochondrial oxygen consumption, suggesting the involvement of a flavoenzyme in this process. Furthermore, we identified the increased oxidation of highly conserved cysteine residues in key proteins such as the epidermal growth factor receptor, filamin A, fibulin-2, and endosialin during the differentiation process. Collectively, this study provides insights into the sources of H<sub>2</sub>O<sub>2</sub> in fibroblasts and highlights the novel redox mechanisms underpinning fibroblast-to-myofibroblast differentiation.https://www.mdpi.com/2076-3921/14/4/486proteomicsthiol oxidationredox signallingfibroblastsmyofibroblaststransforming growth factor β1 |
| spellingShingle | Marzieh Aminzadehanboohi Manousos Makridakis Delphine Rasti Yves Cambet Karl-Heinz Krause Antonia Vlahou Vincent Jaquet Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast Differentiation Antioxidants proteomics thiol oxidation redox signalling fibroblasts myofibroblasts transforming growth factor β1 |
| title | Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast Differentiation |
| title_full | Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast Differentiation |
| title_fullStr | Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast Differentiation |
| title_full_unstemmed | Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast Differentiation |
| title_short | Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast Differentiation |
| title_sort | redox mechanisms driving skin fibroblast to myofibroblast differentiation |
| topic | proteomics thiol oxidation redox signalling fibroblasts myofibroblasts transforming growth factor β1 |
| url | https://www.mdpi.com/2076-3921/14/4/486 |
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