Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts
In fibrosis remodelling of ECM leads to changes in composition and stiffness. Such changes can have a major impact on cell functions including proliferation, secretory profile and differentiation. Several studies have reported that fibrosis is characterised by increased senescence and accumulating e...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2022.989169/full |
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| author | Kaj E. C. Blokland Kaj E. C. Blokland Kaj E. C. Blokland Kaj E. C. Blokland Mehmet Nizamoglu Mehmet Nizamoglu Habibie Habibie Habibie Habibie Habibie Habibie Theo Borghuis Theo Borghuis Michael Schuliga Barbro N. Melgert Barbro N. Melgert Barbro N. Melgert Darryl A. Knight Darryl A. Knight Darryl A. Knight Corry-Anke Brandsma Corry-Anke Brandsma Simon D. Pouwels Simon D. Pouwels Simon D. Pouwels Janette K. Burgess Janette K. Burgess |
| author_facet | Kaj E. C. Blokland Kaj E. C. Blokland Kaj E. C. Blokland Kaj E. C. Blokland Mehmet Nizamoglu Mehmet Nizamoglu Habibie Habibie Habibie Habibie Habibie Habibie Theo Borghuis Theo Borghuis Michael Schuliga Barbro N. Melgert Barbro N. Melgert Barbro N. Melgert Darryl A. Knight Darryl A. Knight Darryl A. Knight Corry-Anke Brandsma Corry-Anke Brandsma Simon D. Pouwels Simon D. Pouwels Simon D. Pouwels Janette K. Burgess Janette K. Burgess |
| author_sort | Kaj E. C. Blokland |
| collection | DOAJ |
| description | In fibrosis remodelling of ECM leads to changes in composition and stiffness. Such changes can have a major impact on cell functions including proliferation, secretory profile and differentiation. Several studies have reported that fibrosis is characterised by increased senescence and accumulating evidence suggests that changes to the ECM including altered composition and increased stiffness may contribute to premature cellular senescence. This study investigated if increased stiffness could modulate markers of senescence and/or fibrosis in primary human lung fibroblasts. Using hydrogels representing stiffnesses that fall within healthy and fibrotic ranges, we cultured primary fibroblasts from non-diseased lung tissue on top of these hydrogels for up to 7 days before assessing senescence and fibrosis markers. Fibroblasts cultured on stiffer (±15 kPa) hydrogels showed higher Yes-associated protein-1 (YAP) nuclear translocation compared to soft hydrogels. When looking at senescence-associated proteins we also found higher secretion of receptor activator of nuclear factor kappa-B ligand (RANKL) but no change in transforming growth factor-β1 (TGF-β1) or connective tissue growth factor (CTGF) expression and higher decorin protein deposition on stiffer matrices. With respect to genes associated with fibrosis, fibroblasts on stiffer hydrogels compared to soft had higher expression of smooth muscle alpha (α)-2 actin (ACTA2), collagen (COL) 1A1 and fibulin-1 (Fbln1) and higher Fbln1 protein deposition after 7 days. Our results show that exposure of lung fibroblasts to fibrotic stiffness activates genes and secreted factors that are part of fibrotic responses and part of the Senescence-associated secretory phenotype (SASP). This overlap may contribute to the creation of a feedback loop whereby fibroblasts create a perpetuating cycle reinforcing progression of a fibrotic response. |
| first_indexed | 2024-04-12T14:20:10Z |
| format | Article |
| id | doaj.art-db3ba90f7fe2417081a0e39944672a7b |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-04-12T14:20:10Z |
| publishDate | 2022-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Pharmacology |
| spelling | doaj.art-db3ba90f7fe2417081a0e39944672a7b2022-12-22T03:29:37ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122022-11-011310.3389/fphar.2022.989169989169Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblastsKaj E. C. Blokland0Kaj E. C. Blokland1Kaj E. C. Blokland2Kaj E. C. Blokland3Mehmet Nizamoglu4Mehmet Nizamoglu5Habibie Habibie6Habibie Habibie7Habibie Habibie8Theo Borghuis9Theo Borghuis10Michael Schuliga11Barbro N. Melgert12Barbro N. Melgert13Barbro N. Melgert14Darryl A. Knight15Darryl A. Knight16Darryl A. Knight17Corry-Anke Brandsma18Corry-Anke Brandsma19Simon D. Pouwels20Simon D. Pouwels21Simon D. Pouwels22Janette K. Burgess23Janette K. Burgess24University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, NetherlandsUniversity of Newcastle, School of Biomedical Sciences and Pharmacy, Callaghan, NSW, AustraliaNational Health and Medical Research Council Centre of Research Excellence in Pulmonary Fibrosis, Sydney, NSW, AustraliaUniversity of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, NetherlandsUniversity of Groningen, Department of Molecular Pharmacology, Groningen Research Institute of Pharmacy, Groningen, NetherlandsHasanuddin University, Faculty of Pharmacy, Makassar, IndonesiaUniversity of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, NetherlandsUniversity of Newcastle, School of Biomedical Sciences and Pharmacy, Callaghan, NSW, AustraliaUniversity of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, NetherlandsUniversity of Groningen, Department of Molecular Pharmacology, Groningen Research Institute of Pharmacy, Groningen, NetherlandsUniversity of Newcastle, School of Biomedical Sciences and Pharmacy, Callaghan, NSW, AustraliaNational Health and Medical Research Council Centre of Research Excellence in Pulmonary Fibrosis, Sydney, NSW, AustraliaProvidence Health Care Research Institute, Vancouver, BC, CanadaUniversity of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Pulmonology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, NetherlandsIn fibrosis remodelling of ECM leads to changes in composition and stiffness. Such changes can have a major impact on cell functions including proliferation, secretory profile and differentiation. Several studies have reported that fibrosis is characterised by increased senescence and accumulating evidence suggests that changes to the ECM including altered composition and increased stiffness may contribute to premature cellular senescence. This study investigated if increased stiffness could modulate markers of senescence and/or fibrosis in primary human lung fibroblasts. Using hydrogels representing stiffnesses that fall within healthy and fibrotic ranges, we cultured primary fibroblasts from non-diseased lung tissue on top of these hydrogels for up to 7 days before assessing senescence and fibrosis markers. Fibroblasts cultured on stiffer (±15 kPa) hydrogels showed higher Yes-associated protein-1 (YAP) nuclear translocation compared to soft hydrogels. When looking at senescence-associated proteins we also found higher secretion of receptor activator of nuclear factor kappa-B ligand (RANKL) but no change in transforming growth factor-β1 (TGF-β1) or connective tissue growth factor (CTGF) expression and higher decorin protein deposition on stiffer matrices. With respect to genes associated with fibrosis, fibroblasts on stiffer hydrogels compared to soft had higher expression of smooth muscle alpha (α)-2 actin (ACTA2), collagen (COL) 1A1 and fibulin-1 (Fbln1) and higher Fbln1 protein deposition after 7 days. Our results show that exposure of lung fibroblasts to fibrotic stiffness activates genes and secreted factors that are part of fibrotic responses and part of the Senescence-associated secretory phenotype (SASP). This overlap may contribute to the creation of a feedback loop whereby fibroblasts create a perpetuating cycle reinforcing progression of a fibrotic response.https://www.frontiersin.org/articles/10.3389/fphar.2022.989169/fullcellular senescencestiffnessGelMA hydrogelsfibrosisSASP |
| spellingShingle | Kaj E. C. Blokland Kaj E. C. Blokland Kaj E. C. Blokland Kaj E. C. Blokland Mehmet Nizamoglu Mehmet Nizamoglu Habibie Habibie Habibie Habibie Habibie Habibie Theo Borghuis Theo Borghuis Michael Schuliga Barbro N. Melgert Barbro N. Melgert Barbro N. Melgert Darryl A. Knight Darryl A. Knight Darryl A. Knight Corry-Anke Brandsma Corry-Anke Brandsma Simon D. Pouwels Simon D. Pouwels Simon D. Pouwels Janette K. Burgess Janette K. Burgess Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts Frontiers in Pharmacology cellular senescence stiffness GelMA hydrogels fibrosis SASP |
| title | Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts |
| title_full | Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts |
| title_fullStr | Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts |
| title_full_unstemmed | Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts |
| title_short | Substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts |
| title_sort | substrate stiffness engineered to replicate disease conditions influence senescence and fibrotic responses in primary lung fibroblasts |
| topic | cellular senescence stiffness GelMA hydrogels fibrosis SASP |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2022.989169/full |
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