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Karolien Paul Maria Hollanders, Tine Van Bergen, Evelien Vandewalle, Karolien Castermans, Nele Kindt, Lieve K M Moons, Ingeborg Stalmans; THE EFFECT OF A LOCALLY ACTING ROCK INHIBITOR AMA0428 IN A MODEL OF WET AGE-RELATED MACULAR DEGENERATION. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1963.
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Rho kinase (ROCK) is associated with VEGF-driven angiogenesis, and is involved in inflammation and fibrosis. Therefore, the in vitro and in vivo effect of a locally-acting ROCK inhibitor, AMA0428, in the pathogenesis of wet age-related macular degeneration was studied.
The in vitro effect of AMA0428 (0.5 -5µM) on human brain microvascular endothelial cells (HBMEC), human brain microvascular pericytes (HBVP) and human tenon fibroblasts (HTF) was determined by measuring: cell viability (WST-1), apoptosis (caspase 3/7) and two migration assays (scratch and under-agarose). The in vivo response of AMA0428 was investigated using a laser-induced choroidal neovascularization (CNV) mouse model. Intravitreal injections were given after laser treatment on day 0, 4, 10 and 20 with AMA0428 (0.1, 1, 10, 100 and 1000ng), murine anti-VEGFR antibody (DC101; 6.2 µg) or placebo (vehicle) (10 mice per group). Treatment outcome was blindly assessed by analysis of inflammation (CD45), angiogenesis (FITC-dextran), vessel leakage (Texas Red-conjugated Dextran and FITC-labeled lectin) and fibrosis (Sirius Red).
A dose-dependent reduction of proliferation and VEGF-induced migration of HBMEC and HTF was measured after AMA0428 administration in vitro (P<0.05). No significant effect was seen on HBVP proliferation, however, analysis of the scratch and under-agarose assays showed increased migration and pericyte recruitment (P<0.05). There was no significant apoptosis induction at all concentrations tested (P>0.05). AMA0428 (0.1 - 1000ng) dose-dependently reduced angiogenesis 2 weeks after CNV induction, (P<0.05), this was associated with a decrease in vessel leakage (P<0.05). These data are in line with those of DC101 (P>0.05). In addition, the highest dose of AMA0428 inhibited inflammation on day 5 by 20% (P<0.05) and collagen deposition on day 30 by 39% (P<0.001) while vehicle and DC101 had no effect on inflammation nor fibrosis.
Our in vitro and in vivo data suggest that targeting ROCK with AMA0428 not only reduces neoangiogenesis (similar to VEGF inhibition), but also blocks inflammation and fibrosis (contrary to VEGF inhibition). These results point to a potential therapeutic benefit of ROCK inhibition in wet AMD.
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