RhoA/ROCK signaling is essential for multiple aspects of VEGF‐mediated angiogenesis

Abstract

ABSTRACT The small GTPase RhoA and its downstream effectors, ROCK1 and ROCK2, regulate a number of cellular processes, including cell motility, proliferation, survival, and permeability. Pharmacological inhibitors of the Rho pathway reportedly block angiogenesis; however, the molecular details of this inhibition are largely unknown. We demonstrate that vascular endothelial growth factor‐A (VEGF) rapidly induces RhoA activation in endothelial cells (ECs). Moreover, the pharmacological inhibition of ROCK1/2 using 10 µM Y‐27632 (the IC 50 for this compound in ECs) strongly disrupts vasculogenesis in pluripotent embryonic stem cell cultures, VEGF‐mediated regenerative angiogenesis in ex vivo retinal explants, and VEGF‐mediated in vitro EC tube formation. Furthermore, using small interfering RNA knockdown and mouse heterozygote knockouts of ROCK1 and ROCK2, we provide data indicating that VEGF‐driven angiogenesis is largely mediated through ROCK2. These data demonstrate that Rho/ROCK signaling is an important mediator in a number of angiogenic processes, including EC migration, survival, and cell permeability, and suggest that Rho/ROCK inhibition may prove useful for the treatment of angiogenesis‐related disorders.—Bryan, B. A., Dennstedt, E., Mitchell, D. C., Walshe, T. E., Noma, K., Loureiro, R., Saint‐Geniez, M., Campaigniac, J.‐P., Liao, J. K., D'Amore, P. A. RhoA/ROCK signaling is essential for multiple aspects of VEGF‐mediated angiogenesis. FASEB J . 24, 3186–3195 (2010). www.fasebj.org