Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis
Fibrosis is a major cause of progressive organ dysfunction in several chronic pulmonary diseases. Rho associated coiled-coil forming kinase (ROCK) has shown to be involved in myofibroblast differentiation driven by altered matrix stiffness in fibrotic state. There are two known ROCK isoforms in huma...
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| Format: | Article |
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American Thoracic Society
2017
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| Online Access: | https://eprints.nottingham.ac.uk/40910/ |
| _version_ | 1848796160915931136 |
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| author | Htwe, Su S. Cha, Byung H. Yue, Kan Khademhosseini, Ali Knox, Alan J. Ghaemmaghami, Amir M. |
| author_facet | Htwe, Su S. Cha, Byung H. Yue, Kan Khademhosseini, Ali Knox, Alan J. Ghaemmaghami, Amir M. |
| author_sort | Htwe, Su S. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Fibrosis is a major cause of progressive organ dysfunction in several chronic pulmonary diseases. Rho associated coiled-coil forming kinase (ROCK) has shown to be involved in myofibroblast differentiation driven by altered matrix stiffness in fibrotic state. There are two known ROCK isoforms in human, ROCK1 (ROKβ) and ROCK2 (ROKα), but specific role of each isoform in myofibroblast differentiation in lung fibrosis remains unknown. To study this, we developed a Gelatin methacryloyl (GelMA) hydrogel based culture system with different stiffness levels relevant to healthy and fibrotic lungs. We have shown that stiff matrix and not soft matrix, can induce myofibroblast differentiation with high αSMA expression. Furthermore, our data confirm that the inhibition of ROCK signalling by a pharmacological inhibitor (i.e. Y27632) attenuates stiffness induced αSMA expression and fibre assembly in myofibroblasts. To assess the role of ROCK isoforms in this process we used siRNA to knock down the expression of each isoform. Our data showed that knocking down either ROCK1 or ROCK2 did not result in a reduction in αSMA expression in myofibroblasts on stiff matrix as opposed to soft matrix where αSMA expression was reduced significantly. Paradoxically, on stiff matrix, the absence of one isoform (particularly ROCK2) exaggerated αSMA expression and led to thick fibre assembly. Moreover complete loss of αSMA fibre assembly was seen only in the absence of both ROCK isoforms suggesting that both isoforms are implicated in this process. Overall our results indicate the differential role of ROCK isoforms in myofibroblast differentiation on soft and stiff matrices. |
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| format | Article |
| id | nottingham-40910 |
| institution | University of Nottingham Malaysia Campus |
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| last_indexed | 2025-11-14T19:43:34Z |
| publishDate | 2017 |
| publisher | American Thoracic Society |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-409102020-05-04T18:47:59Z https://eprints.nottingham.ac.uk/40910/ Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis Htwe, Su S. Cha, Byung H. Yue, Kan Khademhosseini, Ali Knox, Alan J. Ghaemmaghami, Amir M. Fibrosis is a major cause of progressive organ dysfunction in several chronic pulmonary diseases. Rho associated coiled-coil forming kinase (ROCK) has shown to be involved in myofibroblast differentiation driven by altered matrix stiffness in fibrotic state. There are two known ROCK isoforms in human, ROCK1 (ROKβ) and ROCK2 (ROKα), but specific role of each isoform in myofibroblast differentiation in lung fibrosis remains unknown. To study this, we developed a Gelatin methacryloyl (GelMA) hydrogel based culture system with different stiffness levels relevant to healthy and fibrotic lungs. We have shown that stiff matrix and not soft matrix, can induce myofibroblast differentiation with high αSMA expression. Furthermore, our data confirm that the inhibition of ROCK signalling by a pharmacological inhibitor (i.e. Y27632) attenuates stiffness induced αSMA expression and fibre assembly in myofibroblasts. To assess the role of ROCK isoforms in this process we used siRNA to knock down the expression of each isoform. Our data showed that knocking down either ROCK1 or ROCK2 did not result in a reduction in αSMA expression in myofibroblasts on stiff matrix as opposed to soft matrix where αSMA expression was reduced significantly. Paradoxically, on stiff matrix, the absence of one isoform (particularly ROCK2) exaggerated αSMA expression and led to thick fibre assembly. Moreover complete loss of αSMA fibre assembly was seen only in the absence of both ROCK isoforms suggesting that both isoforms are implicated in this process. Overall our results indicate the differential role of ROCK isoforms in myofibroblast differentiation on soft and stiff matrices. American Thoracic Society 2017-06-01 Article PeerReviewed Htwe, Su S., Cha, Byung H., Yue, Kan, Khademhosseini, Ali, Knox, Alan J. and Ghaemmaghami, Amir M. (2017) Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis. American Journal of Respiratory Cell and Molecular Biology, 56 (6). ISSN 1535-4989 http://www.atsjournals.org/doi/10.1165/rcmb.2016-0306OC doi:10.1165/rcmb.2016-0306OC doi:10.1165/rcmb.2016-0306OC |
| spellingShingle | Htwe, Su S. Cha, Byung H. Yue, Kan Khademhosseini, Ali Knox, Alan J. Ghaemmaghami, Amir M. Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis |
| title | Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis |
| title_full | Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis |
| title_fullStr | Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis |
| title_full_unstemmed | Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis |
| title_short | Role of ROCK isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis |
| title_sort | role of rock isoforms in regulation of stiffness induced myofibroblast differentiation in lung fibrosis |
| url | https://eprints.nottingham.ac.uk/40910/ https://eprints.nottingham.ac.uk/40910/ https://eprints.nottingham.ac.uk/40910/ |