title: The effect of AMA0428, a novel and potent ROCK inhibitor, in a model of neovascular age-related macular degeneration.
abstract: OBJECTIVE
  Rho kinase (ROCK) is associated with VEGF-driven angiogenesis, as well as with inflammation and fibrosis. Therefore, the effect of AMA0428, a novel ROCK inhibitor, was studied in these processes, which highly contribute to the pathogenesis of neovascular AMD.
  
  
  METHODS
  The effect of AMA0428 (0.5-5.0 μM) on human umbilical vein endothelial cells (HUVECs), human brain microvascular endothelial cells (HBMECs), and human brain microvascular pericytes (HBVPs) was determined using cell viability (WST-1), apoptosis (caspase 3/7), and migration (scratch and under-agarose) assays. The in vivo response was investigated using a laser-induced choroidal neovascularization (CNV) mouse model, in which intravitreal injections of AMA0428, murine anti-VEGF-R2 mAb (DC101), or placebo was given. Outcome was assessed by analysis of inflammation (CD45), angiogenesis (FITC-dextran), vessel leakage (Texas Red-conjugated Dextran and FITC-labeled lectin) and fibrosis (Sirius Red/Collagen I).
  
  
  RESULTS
  The AMA0428 dose-dependently reduced proliferation and VEGF-induced migration of HUVEC and HBMEC (P < 0.05). No significant effect was seen on HBVP proliferation; however, migration and pericyte recruitment were enhanced (P < 0.05) by AMA0428 administration. There was no apoptosis induction. The AMA0428 significantly reduced CNV and vessel leakage 2 weeks after laser treatment, comparable to DC101. In addition, AMA0428 inhibited inflammation on day 5 by 42% (P < 0.05) and collagen deposition on day 30 by 43% (P < 0. 05), whereas DC101 had no effect on inflammation nor on fibrosis.
  
  
  CONCLUSIONS
  The results suggest that targeting ROCK with AMA0428 not only reduces neoangiogenesis, but also blocks inflammation and fibrosis (contrary to VEGF suppression). These results point to a potential therapeutic benefit of ROCK inhibition in neovascular AMD.
authors: KU Leuven-University of Leuven, Department of Neurosciences, Laboratory of Ophthalmology, Leuven, Belgium.
authors: Karolien
authors: K
authors: Hollanders
authors: KU Leuven-University of Leuven, Department of Neurosciences, Laboratory of Ophthalmology, Leuven, Belgium.
authors: Tine
authors: T
authors: Van Bergen
authors: Amakem Therapeutics, Diepenbeek, Belgium.
authors: Nele
authors: N
authors: Kindt
authors: Amakem Therapeutics, Diepenbeek, Belgium.
authors: Karolien
authors: K
authors: Castermans
authors: Amakem Therapeutics, Diepenbeek, Belgium.
authors: Dirk
authors: D
authors: Leysen
authors: KU Leuven-University of Leuven, Department of Neurosciences, Laboratory of Ophthalmology, Leuven, Belgium KU Leuven-University of Leuven, University Hospitals Leuven, Department of Ophthalmology, Leuven, Belgium.
authors: Evelien
authors: E
authors: Vandewalle
authors: KU Leuven-University of Leuven, Department of Neurosciences, Laboratory of Ophthalmology, Leuven, Belgium KU Leuven-University of Leuven, Department of Biology, KU Leuven, Leuven, Belgium.
authors: Lieve
authors: L
authors: Moons
authors: KU Leuven-University of Leuven, Department of Neurosciences, Laboratory of Ophthalmology, Leuven, Belgium KU Leuven-University of Leuven, University Hospitals Leuven, Department of Ophthalmology, Leuven, Belgium.
authors: Ingeborg
authors: I
authors: Stalmans
mesh_terms: D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000627:therapeutic use*; D006801:Humans; D008268:Macular Degeneration / Q000188:drug therapy* / Q000209:etiology / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015861:Retinal Neovascularization / Q000150:complications* / Q000209:etiology / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
chemical_list: D004791:Enzyme Inhibitors; D042461:Vascular Endothelial Growth Factor A; D054460:rho-Associated Kinases
keywords: age-related macular degeneration; anti-vascular endothelial growth factor; rho kinase inhibition