Involvement of RhoA/Rho-Associated Kinase Signal Transduction Pathway in Dexamethasone-Induced Alterations in Aqueous Outflow

Fujimoto, Tomokazu; Inoue, Toshihiro; Kameda, Takuro; Kasaoka, N.; Inoue-Mochita, Miyuki; Tsuboi, Naoko; Tanihara, Hidenobu

doi:10.1167/iovs.12-9989

Cited by 59 publications

(59 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, with the use of in vitro experiments using cultured trabecular meshwork and Schlemm canal endothelial cells, cytoskeletal redistribution and changes in cell-cell interactions in trabecular meshwork and Schlemm canal endothelial cells have been reported. 4,6,13 Ripasudil has selective rho kinase inhibitory effects and IOP-lowering and outflow facilityincreasing effects in animal eyes. 5 Because these mechanisms of rho kinase inhibitors (including ripasudil) are different from the known IOP-lowering mechanisms for other antiglaucoma medications, additive IOP-lowering effects had been expected.…”

Section: Discussionmentioning

confidence: 99%

Additive Intraocular Pressure–Lowering Effects of the Rho Kinase Inhibitor Ripasudil (K-115) Combined With Timolol or Latanoprost

et al. 2015

Self Cite

View full text Add to dashboard Cite

for the K-115 Clinical Study Group IMPORTANCE Ripasudil hydrochloride hydrate (K-115), a novel rho kinase inhibitor, provides statistically significant intraocular pressure (IOP)-lowering effects and has a tolerable safety profile. However, no studies have evaluated ripasudil combined with β-blockers and prostaglandin analogues.OBJECTIVE To evaluate the additive IOP-lowering effects and the safety of ripasudil, 0.4%, combined with timolol, 0.5%, or latanoprost, 0.005%, in patients with primary open-angle glaucoma or ocular hypertension. DESIGN, SETTING, AND PARTICIPANTSWe conducted 2, multicenter, randomized, double-masked, parallel group comparison studies of ripasudil-timolol and ripasudillatanoprost in 29 and 36 Japanese clinical centers, respectively. Analyses were performed on an intention-treat-treat basis. After appropriate run-in periods with timolol or latanoprost, 208 and 205 patients whose IOP levels were 18 mm Hg or higher were enrolled in the ripasudil-timolol and ripasudil-latanoprost groups, respectively. Enrollment began December 1, 2011, and follow-up was completed on September 7, 2012, in the ripasudil-timolol study. Enrollment began December 1, 2011, and follow-up was completed on September 27, 2012, in the ripasudil-latanoprost study.INTERVENTIONS Patients were subdivided into 2 groups in each study and were treated with ripasudil or placebo twice daily for 8 weeks. MAIN OUTCOMES AND MEASURESThe IOP reductions in the ripasudil and placebo groups were analyzed with a repeated-measures analysis of variance model at weeks 4, 6, and 8, at trough (before instillation [9 AM]) and peak (2 hours after instillation [11 AM]) levels. RESULTSIn the ripasudil-timolol study, the mean IOP reductions from baseline in the ripasudil and placebo groups were −2.4 and −1.5 mm Hg at 9 AM for a difference of 0.9 mm Hg (95% CI, 0.4-1.3 mm Hg; P < .001) and −2.9 and −1.3 mm Hg at 11 AM for a difference of 1.6 mm Hg (95% CI, 1.1-2.1 mm Hg; P < .001), respectively. In the ripasudil-latanoprost study, those IOP reductions were −2.2 and −1.8 mm Hg at 9 AM for a difference of 0.4 mm Hg (95% CI, −0.0 to 0.9 mm Hg; P = .06) and −3.2 and −1.8 mm Hg at 11 AM for a difference of 1.4 mm Hg (95% CI, 0.9-1.9 mm Hg; P < .001), respectively. The most frequently reported adverse event was conjunctival hyperemia, which was mild and in most cases resolved without treatment before the next instillation.CONCLUSIONS AND RELEVANCE These clinical trials found additive IOP-lowering effects of ripasudil from placebo at trough and peak levels in combination with timolol and at peak level in combination with latanoprost. However, a definitive difference in the addition of placebo to latanoprost was not identified in the trough level.

show abstract

Section: Discussionmentioning

confidence: 99%

Additive Intraocular Pressure–Lowering Effects of the Rho Kinase Inhibitor Ripasudil (K-115) Combined With Timolol or Latanoprost

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…Primary monkey TM and SCE cells were isolated from the eyes according to a previously described method [22, 23]. Briefly, primary monkey TM and SCE cells were cultured in Dulbecco’s modified Eagle medium (DMEM; WAKO Pure Chemical Industries, Osaka, Japan) in the presence of 10% fetal bovine serum (FBS), glutamine (2 mM), penicillin (100 U/mL), streptomycin (100 μg/mL), and amphotericin B (0.5 μg/mL) at 37°C in 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Vascular Endothelial Growth Factor-A Increases the Aqueous Humor Outflow Facility

et al. 2016

Self Cite

View full text Add to dashboard Cite

PurposeAnti-vascular endothelial growth factor (VEGF) antibody therapy is an effective treatment for ocular angiogenesis. Although the intraocular pressure of some patients increases after anti-VEGF therapy, the effects of VEGF-A on the aqueous humor outflow pathway remain unknown. This study investigated the effects of VEGF-A on the aqueous humor outflow pathway.MethodsWe used human recombinant VEGF121 and VEGF165. Trabecular meshwork (TM) and Schlemm’s canal endothelial (SCE) cells were isolated from the eyes of cynomolgus monkeys. Expression of mRNA coding four VEGF receptors, VEGFR1 (FLT1), VEGFR2 (KDR), neuropilin-1, and neuropilin-2, was examined by RT-PCR. To evaluate the permeability of cell monolayers, we measured transendothelial electrical resistance (TEER). The outflow facility was measured in perfused porcine anterior segment organ cultures treated with 30 ng/mL VEGF121 for 48 h.ResultsFour VEGF-A-related receptor mRNAs were expressed in TM and SCE cells. The TEER of TM cells was not significantly affected by VEGF121 or VEGF165 treatment. In contrast, the TEER of SCE cells was significantly lower 48 h after treatment with 30 ng/mL VEGF121 to 69.4 ± 12.2% of baseline (n = 10), which was a significant difference compared with the control (P = 0.0001). VEGF165 (30 ng/mL) decreased the TEER of SCE cells at 48 h after treatment to 72.3 ± 14.1% compared with the baseline (n = 10), which was not a significant difference compared with the control (P = 0.0935). Ki8751, a selective VEGFR2 inhibitor, completely suppressed the effect of VEGF121 on SCE cell permeability, although ZM306416, a selective VEGFR1 inhibitor, did not affect the VEGF121-induced decrease in TEER. Perfusion with 30 ng/mL of VEGF121 for 48 h significantly increased the outflow facility compared with the control (47.8 ± 28.5%, n = 5, P = 0.013).ConclusionsThese results suggest that VEGF-A may regulate the conventional aqueous outflow of SCE cells through VEGFR2.

show abstract

“…37 Briefly, cells were fixed with 4% paraformaldehyde (PFA), permeabilized with 0.5% Triton X-100, blocked with 10% fetal bovine serum, and then incubated overnight with monoclonal antibodies against YAP/TAZ (cat. no.…”

Section: Immunocytochemistrymentioning

confidence: 99%

YAP/TAZ Are Essential for TGF-β2–Mediated Conjunctival Fibrosis

Futakuchi

Inoue

Wei

et al. 2018

Invest. Ophthalmol. Vis. Sci.

Self Cite

View full text Add to dashboard Cite

PURPOSE. To investigate the roles of Yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ), the major effector molecules of the Hippo pathway, in TGFb2-mediated conjunctival fibrosis.METHODS. Primary human conjunctival fibroblasts were treated with TGF-b2. The expression of YAP/TAZ was examined by Western blot analyses and immunocytochemistry. The expression of fibrotic proteins and genes were evaluated by Western blot analyses and quantitative real-time PCR, respectively. The effects of YAP/TAZ on fibrotic changes were examined by knockdown experiments and the YAP/TAZ inhibitor, verteporfin.RESULTS. TGF-b2 stabilized YAP/TAZ and subsequently activated Smad2/3, which led to the transcription of fibrotic genes in human primary conjunctival fibroblasts. These fibrotic genes were differently regulated by YAP/TAZ. Notably, a-smooth muscle actin, fibronectin, collagen I, and collagen IV were primarily regulated by YAP. In contrast, CCN family proteins (CTGF and CYR61) depended on both YAP and TAZ. Mechanistically, YAP/TAZ were located in close proximity to Smad2/3, and in particular, YAP was required for TGF-b2-mediated phosphorylation and the nuclear translocation of Smad2/3. Furthermore, a YAP/TAZ inhibitor markedly suppressed TGF-b2-mediated fibrotic changes in conjunctival fibroblasts.CONCLUSIONS. YAP/TAZ acted as a molecular hub of TGF-b2 signaling in a cellular model of conjunctival fibrosis. Moreover, verteporfin, a YAP/TAZ inhibitor exerted potent antifibrosis effects by suppressing TGF-b2-YAP/TAZ-Smad signaling. Our study highlights YAP/TAZ as essential regulators of conjunctival fibrosis and shows that inhibition of YAP/TAZ might potentially improve the outcomes of glaucoma filtration surgery.

show abstract

Involvement of RhoA/Rho-Associated Kinase Signal Transduction Pathway in Dexamethasone-Induced Alterations in Aqueous Outflow

Cited by 59 publications

References 30 publications

Additive Intraocular Pressure–Lowering Effects of the Rho Kinase Inhibitor Ripasudil (K-115) Combined With Timolol or Latanoprost

Additive Intraocular Pressure–Lowering Effects of the Rho Kinase Inhibitor Ripasudil (K-115) Combined With Timolol or Latanoprost

Vascular Endothelial Growth Factor-A Increases the Aqueous Humor Outflow Facility

YAP/TAZ Are Essential for TGF-β2–Mediated Conjunctival Fibrosis

Contact Info

Product

Resources

About