ROCK 1 and 2 affect the spatial architecture of 3D spheroids derived from human corneal stromal fibroblasts in different manners
Abstract The objective of the current study was to examine the roles of ROCK1 and 2 on the spatial architecture of human corneal stroma. We examined the effects of a pan-ROCK inhibitor (pan-ROCK-i), ripasudil, and a ROCK2 inhibitor (ROCK2-i), KD025 on the expression of genes that encode for ECM prot...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-11407-1 |
| _version_ | 1828785452831735808 |
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| author | Yosuke Ida Araya Umetsu Masato Furuhashi Megumi Watanabe Yuri Tsugeno Soma Suzuki Fumihito Hikage Hiroshi Ohguro |
| author_facet | Yosuke Ida Araya Umetsu Masato Furuhashi Megumi Watanabe Yuri Tsugeno Soma Suzuki Fumihito Hikage Hiroshi Ohguro |
| author_sort | Yosuke Ida |
| collection | DOAJ |
| description | Abstract The objective of the current study was to examine the roles of ROCK1 and 2 on the spatial architecture of human corneal stroma. We examined the effects of a pan-ROCK inhibitor (pan-ROCK-i), ripasudil, and a ROCK2 inhibitor (ROCK2-i), KD025 on the expression of genes that encode for ECM proteins including collagen (COL) 1, 4, 6, and fibronectin (FN), their regulators, a tissue inhibitor of metalloproteinase (TIMP) 1–4, matrix metalloproteinase (MMP) 2, 9 and 14, and ER stress-related factors of two- and three-dimensional (2D and 3D) cultures of human corneal stroma fibroblasts (HCSFs), and the physical properties of 3D HCSF spheroids. A gene expression analysis using ROCK-is indicated that KD025 (ROCK2 selective ROCK inhibitor) induced more significant changes than Rip (ripasudil, pan-ROCK inhibitor), suggesting that ROCK2 might be more extensively involved in the metabolism of ECM proteins and cell architectures of the 2D cultured HCSFs than ROCK1. In terms of the physical properties, size and stiffness of the 3D HCSFs spheroids, Rip caused a significant enlargement and this enhancement was concentration-dependent while KD025 also exerted a similar but less pronounced effect. In contrast, Rip and KD025 modulated physical stiffness differently, in that Rip caused a substantial decrease and KD025 caused an increase. Such diverse effects between Rip and KD025 were also observed for the gene expressions of ECM proteins, their regulators, and ER-stress related factors. The findings presented herein suggest that the ROCK1 and 2 influence the spatial architecture of 3D HCFS spheroids in different manners. |
| first_indexed | 2024-12-11T23:51:34Z |
| format | Article |
| id | doaj.art-bbbcf84cb01943108006513b6eb48188 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-11T23:51:34Z |
| publishDate | 2022-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-bbbcf84cb01943108006513b6eb481882022-12-22T00:45:28ZengNature PortfolioScientific Reports2045-23222022-05-0112111310.1038/s41598-022-11407-1ROCK 1 and 2 affect the spatial architecture of 3D spheroids derived from human corneal stromal fibroblasts in different mannersYosuke Ida0Araya Umetsu1Masato Furuhashi2Megumi Watanabe3Yuri Tsugeno4Soma Suzuki5Fumihito Hikage6Hiroshi Ohguro7Departments of Ophthalmology, Sapporo Medical University School of MedicineDepartments of Ophthalmology, Sapporo Medical University School of MedicineCardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of MedicineDepartments of Ophthalmology, Sapporo Medical University School of MedicineDepartments of Ophthalmology, Sapporo Medical University School of MedicineDepartments of Ophthalmology, Sapporo Medical University School of MedicineDepartments of Ophthalmology, Sapporo Medical University School of MedicineDepartments of Ophthalmology, Sapporo Medical University School of MedicineAbstract The objective of the current study was to examine the roles of ROCK1 and 2 on the spatial architecture of human corneal stroma. We examined the effects of a pan-ROCK inhibitor (pan-ROCK-i), ripasudil, and a ROCK2 inhibitor (ROCK2-i), KD025 on the expression of genes that encode for ECM proteins including collagen (COL) 1, 4, 6, and fibronectin (FN), their regulators, a tissue inhibitor of metalloproteinase (TIMP) 1–4, matrix metalloproteinase (MMP) 2, 9 and 14, and ER stress-related factors of two- and three-dimensional (2D and 3D) cultures of human corneal stroma fibroblasts (HCSFs), and the physical properties of 3D HCSF spheroids. A gene expression analysis using ROCK-is indicated that KD025 (ROCK2 selective ROCK inhibitor) induced more significant changes than Rip (ripasudil, pan-ROCK inhibitor), suggesting that ROCK2 might be more extensively involved in the metabolism of ECM proteins and cell architectures of the 2D cultured HCSFs than ROCK1. In terms of the physical properties, size and stiffness of the 3D HCSFs spheroids, Rip caused a significant enlargement and this enhancement was concentration-dependent while KD025 also exerted a similar but less pronounced effect. In contrast, Rip and KD025 modulated physical stiffness differently, in that Rip caused a substantial decrease and KD025 caused an increase. Such diverse effects between Rip and KD025 were also observed for the gene expressions of ECM proteins, their regulators, and ER-stress related factors. The findings presented herein suggest that the ROCK1 and 2 influence the spatial architecture of 3D HCFS spheroids in different manners.https://doi.org/10.1038/s41598-022-11407-1 |
| spellingShingle | Yosuke Ida Araya Umetsu Masato Furuhashi Megumi Watanabe Yuri Tsugeno Soma Suzuki Fumihito Hikage Hiroshi Ohguro ROCK 1 and 2 affect the spatial architecture of 3D spheroids derived from human corneal stromal fibroblasts in different manners Scientific Reports |
| title | ROCK 1 and 2 affect the spatial architecture of 3D spheroids derived from human corneal stromal fibroblasts in different manners |
| title_full | ROCK 1 and 2 affect the spatial architecture of 3D spheroids derived from human corneal stromal fibroblasts in different manners |
| title_fullStr | ROCK 1 and 2 affect the spatial architecture of 3D spheroids derived from human corneal stromal fibroblasts in different manners |
| title_full_unstemmed | ROCK 1 and 2 affect the spatial architecture of 3D spheroids derived from human corneal stromal fibroblasts in different manners |
| title_short | ROCK 1 and 2 affect the spatial architecture of 3D spheroids derived from human corneal stromal fibroblasts in different manners |
| title_sort | rock 1 and 2 affect the spatial architecture of 3d spheroids derived from human corneal stromal fibroblasts in different manners |
| url | https://doi.org/10.1038/s41598-022-11407-1 |
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