Inhibition of Rho-dependent kinases ROCK I/II activates VEGF-driven retinal neovascularization and sprouting angiogenesis

Am J Physiol Heart Circ Physiol. 2009 Mar;296(3):H893-9. doi: 10.1152/ajpheart.01038.2008. Epub 2009 Jan 30.

Abstract

Vascular endothelial growth factor (VEGF) is an endothelial-specific growth factor that activates the small GTPase RhoA. While the role of RhoA for VEGF-driven endothelial migration and angiogenesis has been studied in detail, the function of its target proteins, the Rho-dependent kinases ROCK I and II, are controversially discussed. Using the mouse model of oxygen-induced proliferative retinopathy, ROCK I/II inhibition by H-1152 resulted in increased angiogenesis. This enhanced angiogenesis, however, was completely blocked by the VEGF-receptor antagonist PTK787/ZK222584. Loss-of-function experiments in endothelial cells revealed that inhibition of ROCK I/II using the pharmacological inhibitor H-1152 and ROCK I/II-specific small-interfering RNAs resulted in a rise of VEGF-driven sprouting angiogenesis. These functional data were biochemically substantiated by showing an enhanced VEGF-receptor kinase insert domain receptor phosphorylation and extracellular signal-regulated kinase 1/2 activation after inhibition of ROCK I/II. Thus our data identify that the inhibition of Rho-dependent kinases ROCK I/II activates angiogenesis both, in vitro and in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / analogs & derivatives*
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / pharmacology
  • Angiogenesis Inducing Agents / pharmacology*
  • Angiogenesis Inhibitors / pharmacology
  • Animals
  • Animals, Newborn
  • Cells, Cultured
  • Disease Models, Animal
  • Dose-Response Relationship, Drug
  • Endothelial Cells / drug effects
  • Endothelial Cells / enzymology
  • Humans
  • Mice
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Neovascularization, Physiologic / drug effects*
  • Oxygen
  • Phthalazines / pharmacology
  • Protein Kinase Inhibitors / pharmacology*
  • Pyridines / pharmacology
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Receptors, Vascular Endothelial Growth Factor / antagonists & inhibitors
  • Receptors, Vascular Endothelial Growth Factor / metabolism
  • Retinal Neovascularization / chemically induced
  • Retinal Neovascularization / enzymology
  • Retinal Neovascularization / physiopathology*
  • Vascular Endothelial Growth Factor A / metabolism*
  • rho-Associated Kinases / antagonists & inhibitors*
  • rho-Associated Kinases / genetics
  • rho-Associated Kinases / metabolism

Substances

  • 2-methyl-1-((4-methyl-5-isoquinolinyl)sulfonyl)homopiperazine
  • Angiogenesis Inducing Agents
  • Angiogenesis Inhibitors
  • Phthalazines
  • Protein Kinase Inhibitors
  • Pyridines
  • RNA, Small Interfering
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • vascular endothelial growth factor A, mouse
  • vatalanib
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • Receptors, Vascular Endothelial Growth Factor
  • ROCK1 protein, human
  • ROCK2 protein, human
  • Rock1 protein, mouse
  • Rock2 protein, mouse
  • rho-Associated Kinases
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Oxygen