TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal Fibroblasts
Excessive cross-linking is a major factor in the resistance to the remodelling of the extracellular matrix (ECM) during fibrotic progression. The role of TGFβ signalling in impairing ECM remodelling has been demonstrated in various fibrotic models. We hypothesised that increased ECM cross-linking by...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/3/984 |
| _version_ | 1797409560247402496 |
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| author | Mariya E. Semkova J. Justin Hsuan |
| author_facet | Mariya E. Semkova J. Justin Hsuan |
| author_sort | Mariya E. Semkova |
| collection | DOAJ |
| description | Excessive cross-linking is a major factor in the resistance to the remodelling of the extracellular matrix (ECM) during fibrotic progression. The role of TGFβ signalling in impairing ECM remodelling has been demonstrated in various fibrotic models. We hypothesised that increased ECM cross-linking by TGFβ contributes to skin fibrosis in Systemic Sclerosis (SSc). Proteomics was used to identify cross-linking enzymes in the ECM of primary human dermal fibroblasts, and to compare their levels following treatment with TGFβ-1. A significant upregulation and enrichment of lysyl-oxidase-like 1, 2 and 4 and transglutaminase 2 were found. Western blotting confirmed the upregulation of lysyl hydroxylase 2 in the ECM. Increased transglutaminase activity in TGFβ-1 treated ECM was revealed from a cell-based assay. We employed a mass spectrometry-based method to identify alterations in the ECM cross-linking pattern caused by TGFβ-1. Cross-linking sites were identified in collagens I and V, fibrinogen and fibronectin. One cross-linking site in fibrinogen alpha was found only in TGFβ-treated samples. In conclusion, we have mapped novel cross-links between ECM proteins and demonstrated that activation of TGFβ signalling in cultured dermal fibroblasts upregulates multiple cross-linking enzymes in the ECM. |
| first_indexed | 2024-03-09T04:16:32Z |
| format | Article |
| id | doaj.art-b156e4b0c0ea4532927b42a64a6f976e |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T04:16:32Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-b156e4b0c0ea4532927b42a64a6f976e2023-12-03T13:53:13ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-01-0122398410.3390/ijms22030984TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal FibroblastsMariya E. Semkova0J. Justin Hsuan1Institute for Liver and Digestive Health, Division of Medicine, University College London, Rowland Hill Street, London NW3 2PF, UKInstitute for Liver and Digestive Health, Division of Medicine, University College London, Rowland Hill Street, London NW3 2PF, UKExcessive cross-linking is a major factor in the resistance to the remodelling of the extracellular matrix (ECM) during fibrotic progression. The role of TGFβ signalling in impairing ECM remodelling has been demonstrated in various fibrotic models. We hypothesised that increased ECM cross-linking by TGFβ contributes to skin fibrosis in Systemic Sclerosis (SSc). Proteomics was used to identify cross-linking enzymes in the ECM of primary human dermal fibroblasts, and to compare their levels following treatment with TGFβ-1. A significant upregulation and enrichment of lysyl-oxidase-like 1, 2 and 4 and transglutaminase 2 were found. Western blotting confirmed the upregulation of lysyl hydroxylase 2 in the ECM. Increased transglutaminase activity in TGFβ-1 treated ECM was revealed from a cell-based assay. We employed a mass spectrometry-based method to identify alterations in the ECM cross-linking pattern caused by TGFβ-1. Cross-linking sites were identified in collagens I and V, fibrinogen and fibronectin. One cross-linking site in fibrinogen alpha was found only in TGFβ-treated samples. In conclusion, we have mapped novel cross-links between ECM proteins and demonstrated that activation of TGFβ signalling in cultured dermal fibroblasts upregulates multiple cross-linking enzymes in the ECM.https://www.mdpi.com/1422-0067/22/3/984transglutaminaseslysyl hydroxylase 2lysyl-oxidase-like enzymesdermal fibroblastsextracellular matrixtransglutaminase cross-linking |
| spellingShingle | Mariya E. Semkova J. Justin Hsuan TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal Fibroblasts International Journal of Molecular Sciences transglutaminases lysyl hydroxylase 2 lysyl-oxidase-like enzymes dermal fibroblasts extracellular matrix transglutaminase cross-linking |
| title | TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal Fibroblasts |
| title_full | TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal Fibroblasts |
| title_fullStr | TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal Fibroblasts |
| title_full_unstemmed | TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal Fibroblasts |
| title_short | TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal Fibroblasts |
| title_sort | tgfβ 1 induced cross linking of the extracellular matrix of primary human dermal fibroblasts |
| topic | transglutaminases lysyl hydroxylase 2 lysyl-oxidase-like enzymes dermal fibroblasts extracellular matrix transglutaminase cross-linking |
| url | https://www.mdpi.com/1422-0067/22/3/984 |
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