Generation and characterization of a recombinant adenovirus expressing vascular endothelial growth factor for studies of neovascularization in the eye

There is accumulating evidence that an increased expression of vascular end othelial growth factor from retinal pigment epithelial cells may be importa nt in choroidal neovascularization, in vivo studies have demonstrated that subretinal injection of recombinant adenovirus vectors produces long-term t ransgene expression specifically within retinal pigment epithelial cells. A recombinant adenovirus encoding of vascular endothelial growth factor (Ad. RSV.VEGF) was therefore produced and characterized in order to determine wh ether an upregulation of vascular endothelial growth factor expression is s ufficient to induce choroidal neovascularization. Ad.RSV.VEGF was produced by homologous recombination and its identity confirmed by rertriction enzym e analysis. Ad.RSV.VEGF was characterized in vitro by the transduction of c ultured retinal pigment epithelial cells. The in vitro characterization con firmed vascular endothelial growth factor mRNA and protein expression from Ad.RSV.VEGF and demonstrated the biological activity of the vascular endoth elial growth factor protein. A preliminary in vivo study suggested that the subretinal injection of Ad.RSV.VEGF induced vascular leakage.