gene therapy using siena technology

为了评估基因治疗方法治疗脉络膜新生血管（CNV）的可行性，我们生成了一种腺相关病毒8型载体（AAV8），该载体编码靶向血管内皮生长因子（VEGF）的小干扰RNA (siRNA)，并确定了其抑制血管生成的能力。我们制作了AAV8/SmVEGF-2。我们将AAV8/SmVEGF-2或对照载体（AAV8/GFP）注入C57BL/6小鼠视网膜下间隙。取眼进行CNV表面积平装分析。与AAV8/GFP相比，视网膜下递送AAV8/SmVEGF-2显著降低激光病变处的CNV。使用ELISA，我们发现在AAV8/SmVEGF-2治疗的眼睛中，VEGF水平降低了约一半。这些结果表明siRNA-VEGF可以在视网膜上表达，并且通过使用稳定的aav介导的siRNA-VEGF表达可以长期抑制CNV。

To assess the feasibility of a gene therapeutic approach to treating choroidal neovascularization (CNV), we generated an adeno-associated virus type 8 vector (AAV8) encoding a small interfering RNA (siRNA) targeting vascular endothelial growth factor (VEGF), and determined its ability to inhibit angiogenesis. We made AAV8/SmVEGF-2. We injected AAV8/SmVEGF-2 or control vector (AAV8/GFP) into the subretinal space in C57BL/6 mice. The eyes were removed for flat mount analysis of CNV surface area. Subretinal delivery of AAV8/SmVEGF-2 significantly diminished CNV at the laser lesions, as compared to AAV8/GFP Using ELISA, we found that VEGF levels were reduced by approximately half in AAV8/SmVEGF-2 treated eyes. These results suggest that siRNA-VEGF can be expressed across the retina and that long-term suppression of CNV is possible through the use of stable AAV-mediated siRNA-VEGF expression.