Rho-Kinase Inhibition Reduces Myofibroblast Differentiation and Proliferation of Scleral Fibroblasts Induced by Transforming Growth Factor β and Experimental Glaucoma

Pitha, Ian; Oglesby, Ericka; Chow, Amanda; Kimball, Elizabeth; Pease, Mary Ellen; Schaub, Julie; Quigley, Harry A.

doi:10.1167/tvst.7.6.6

Cited by 41 publications

(38 citation statements)

References 39 publications

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“…Gel contraction alleviated by AR12286 was dose-related. These results are similar to findings from a study by Pitha et al [ 35 ]. Three types of ROCK inhibitors, namely, H1152, Y-27632, and fasudil, were applied in scleral fibroblasts.…”

Section: Discussionsupporting

confidence: 93%

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AR12286 Alleviates TGF-β-Related Myofibroblast Transdifferentiation and Reduces Fibrosis after Glaucoma Filtration Surgery

Cheng

Chen

et al. 2020

Molecules

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Scar formation can cause the failure of glaucoma filtration surgery. We investigated the effect of AR12286, a selective Rho-associated kinase inhibitor, on myofibroblast transdifferentiation and intraocular pressure assessment in rabbit glaucoma filtration surgery models. Cell migration and collagen contraction were used to demonstrate the functionality of AR12286-modulated human conjunctival fibroblasts (HConFs). Polymerase chain reaction quantitative analysis was used to determine the effect of AR12286 on the production of collagen Type 1A1 and fibronectin 1. Cell migration and collagen contraction in HConFs were activated by TGF-β1. However, compared with the control group, rabbit models treated with AR12286 exhibited higher reduction in intraocular pressure after filtration surgery, and decreased collagen levels at the wound site in vivo. Therefore, increased α-SMA expression in HConFs induced by TGF-β1 could be inhibited by AR12286, and the production of Type 1A1 collagen and fibronectin 1 in TGF-β1-treated HConFs was inhibited by AR12286. Overall, the stimulation of HConFs by TGF-β1 was alleviated by AR12286, and this effect was mediated by the downregulation of TGF-β receptor-related SMAD signaling pathways. In vivo results indicated that AR12286 thus improves the outcome of filtration surgery as a result of its antifibrotic action in the bleb tissue because AR12286 inhibited the TGF-β receptor-related signaling pathway, suppressing several downstream reactions in myofibroblast transdifferentiation.

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Section: Discussionsupporting

confidence: 93%

“…Compared with treatment using other ROCK inhibitors, AR12286 treatment requires lower concentrations (250 and 500 nM). A study reported that 5 to 10 μM ROCK inhibitors were required for myofibroblast differentiation [ 35 , 41 ]. However, drug concentrations may prove to be a limitation of AR12286.…”

Section: Discussionmentioning

confidence: 99%

AR12286 Alleviates TGF-β-Related Myofibroblast Transdifferentiation and Reduces Fibrosis after Glaucoma Filtration Surgery

Cheng

Chen

et al. 2020

Molecules

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“…Thus, activation of fibroblasts by TGF-is an important and essential step by which fibrosis develops. 26 Previous studies have shown that the incubation of human lung fibroblasts with the Ang-(1-7) agonist, A779, reduced levels of TGF-and collagen type I. 27 Therefore, a direct effect of Ang-(1-7) on myofibroblast could have accounted for the antifibrotic effects observed.…”

Section: Discussionmentioning

confidence: 99%

“…Collagen is produced by interstitials fibroblasts that differentiate in myofibroblasts activated by its binding to TGF‐β. Thus, activation of fibroblasts by TGF‐β is an important and essential step by which fibrosis develops . Previous studies have shown that the incubation of human lung fibroblasts with the Ang‐(1‐7) agonist, A779, reduced levels of TGF‐β and collagen type I .…”

Section: Discussionmentioning

confidence: 99%

Effect of preventive or therapeutic treatment with angiotensin 1–7 in a model of bleomycin-induced lung fibrosis in mice

Rago

Melo

Kraemer

et al. 2019

Journal of Leukocyte Biology

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Idiopathic pulmonary fibrosis is characterized by aberrant fibroblast activation and excessive collagen deposition that may eventually lead to organ dysfunction. Lung fibrosis is frequently observed in cancer patients undergoing bleomycin (BLM) treatment. Therefore, BLM instillation in mice is the most frequent model used to investigate pulmonary fibrosis. Angiotensin 1–7 [Ang‐(1‐7)] is a heptapeptide with anti‐inflammatory and proresolving activity. Here, we studied the effects of preventive and therapeutic oral administration of Ang‐(1‐7) in a model of BLM‐induced lung fibrosis in mice. Male C57Bl/6j mice were instilled with BLM and followed for weight loss and survival or euthanized to examine pulmonary inflammation, fibrosis, and lung function. For preventive treatment, mice were treated with Ang‐(1‐7) 1 h before instillation and then twice daily. We observed that preventive treatment with Ang‐(1‐7) decreased weight loss, inflammation and collagen deposition, increased survival, and ameliorated lung function. Therapeutic treatment with Ang‐(1‐7), starting 3 days after BLM instillation resulted in decreased inflammation, decreased collagen deposition, and ameliorated lung function, although the effects were of lower magnitude than the preventive treatment. Therapeutic treatment with Ang‐(1‐7) starting 7 or 14 days after BLM instillation failed to alter any of the changes observed. Therefore, although oral preventive treatment with Ang‐(1‐7) is effective to decrease pulmonary inflammation, fibrosis, and functional changes induced by BLM, therapeutic effects are much less significant, arguing against its use in patients with chronic fibrosis. It remains to be determined whether other proresolving molecules will have better therapeutic effects in the context of chronic pulmonary fibrosis.

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“…The methods of ROCKi delivery may affect the site and mechanism of action. Repeated topical applications can achieve delivery of ROCKi drugs to both anterior and posterior segments, however, intravitreal, subconjunctival, and suprachoroidal injection methods may be more effective for delivery to posterior segment components, such as RGCs and optic nerve axons (Isobe et al, 2016;Pitha et al, 2018). Thus, we used subconjunctival injection to target TM cells, RGCs, and optic nerve axons.…”

Section: Discussionmentioning

confidence: 99%

Rho-Associated Protein Kinase Inhibitor Treatment Promotes Proliferation and Phagocytosis in Trabecular Meshwork Cells

et al. 2020

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Purpose: Continuous reductions in trabecular meshwork (TM) cellularity inhibit aqueous humor (AH) outflow, which is the main cause of primary open-angle glaucoma. Rhoassociated protein kinase inhibitor (ROCKi) targets the TM to reduce intraocular pressure (IOP) and increase AH outflow facility. However, the underlying mechanisms are not entirely clear. Here, we aimed to investigate the effect of a ROCKi (Y-27632) on TM cell proliferation and phagocytosis. Methods: Immortalized human TM (iHTM) cells, glaucomatous TM (GTM 3) cells, and primary human TM (pTM) cells were cultured and identified. The effects of various concentrations of Y-27632 on F-actin cytoskeleton were assessed using immunofluorescence. Cell proliferation effects were evaluated using a cell counting kit-8 (CCK8), cell counting, and Ki67 immunostaining. Cell phagocytosis was evaluated using immunofluorescence and flow cytometry in immortalized TM cells. C57BL/6J and Tg-MYOC Y437H mice were used to investigate the proliferative effects of Y-27632 on TM cells in vivo. The effect of Y-27632 on IOP was monitored for 2 weeks, and the outflow facility was detected 2 weeks after IOP measurement. TM cells in mice were counted using immunohistochemistry. Results: Y-27632 (100 µM) significantly promoted the proliferation of both immortal TM cells and pTM cells. In GTM 3 cells, phagocytosis was significantly greater in the Y-27632 group than in the control group, nearly reaching the level of phagocytosis in iHTM, as determined using immunofluorescence and flow cytometry. In Tg-MYOC Y437H mice, treatment with Y-27632 significantly decreased IOP and increased outflow facility, which greatly influenced the long-term IOP-lowering effect. The number of TM cells in Tg-MYOC Y437H mice was significantly improved after Y-27632 administration. Conclusion: Y-27632 promoted cell proliferation and phagocytosis of TM cells, and its proliferative effect was demonstrated in a transgenic mouse model. These results revealed a new IOP-lowering mechanism of Y-27632 through effects on TM cells, suggesting the potential for a correlation between TM cellularity and long-term recovery of IOP.

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Rho-Kinase Inhibition Reduces Myofibroblast Differentiation and Proliferation of Scleral Fibroblasts Induced by Transforming Growth Factor β and Experimental Glaucoma

Cited by 41 publications

References 39 publications

AR12286 Alleviates TGF-β-Related Myofibroblast Transdifferentiation and Reduces Fibrosis after Glaucoma Filtration Surgery

AR12286 Alleviates TGF-β-Related Myofibroblast Transdifferentiation and Reduces Fibrosis after Glaucoma Filtration Surgery

Effect of preventive or therapeutic treatment with angiotensin 1–7 in a model of bleomycin-induced lung fibrosis in mice

Rho-Associated Protein Kinase Inhibitor Treatment Promotes Proliferation and Phagocytosis in Trabecular Meshwork Cells

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