Decreased expression of the insulin-like growth factor 1 receptor by ribozyme cleavage.

摘要: PURPOSE Insulin-like growth factor (IGF)-1 and its receptor (IGF-1R) are associated with abnormal retinal neovascularization. Ribozymes were designed that selectively decreased the expression of IGF-1R these ribozymes tested in angiogenesis models vitro vivo. METHODS Two hammerhead cleave human mRNA. The cloned into recombinant adeno-associated viral vectors (rAAV). rAAV constructs transfected endothelial cells (HRECs). mRNA protein levels examined modified Boyden chamber assay used to examine ribozyme effects on cell migration. These injected intravitreally mice determine effect neovascularization a mouse model oxygen-induced retinopathy. RESULTS Relative quantitative RT-PCR analysis showed Rz1 reduced by 40% +/- 10% (P = 0.003), Western blot 41% 5% 4.6 x 10(-5)) reduction protein, confirming this reduces expression. also IGF-1-induced migration 90% 2.9 10(-9)) showing function HRECs. amount preretinal 65% 6% 2.7 10(-5)), as measured average number nuclei per section. CONCLUSIONS studies demonstrate effective at reducing Therefore, an method for studying process may ultimately be gene therapy tools pathologic angiogenesis.