NEW METHODOLOGY FOR VEGF-MEDIATED GENE DELIVERY

该项目的总体目标是开发一种商业上可行的系统，用于选择性地将基因输送到生长中的内皮细胞。内皮细胞的生长是血管生成的基础，血管生成几乎完全发生在病理情况下，并由内皮细胞特异性细胞因子血管内皮生长因子(VEGF)促进。在L阶段，我们证明了一种新的方法的可行性，即血管内皮生长因子介导的基因传递，它消除了生长因子或DNA的直接化学处理。在第二阶段的研究中，我们将构建在血管内皮细胞中表达的转基因，并将优化一个基于血管内皮生长因子的载体，用于在体内血管生成部位向内皮细胞高选择性地运送DNA。为了刺激血管生成，我们提出了一种编码血管内皮生长因子的转基因。为了抑制血管生成，我们将构建三种凋亡诱导蛋白与疱疹病毒蛋白vP22融合的转基因，以确保融合蛋白在细胞间的运输。我们选择的蛋白质是：P53、Proaspase-8a和Granzyme B。实现这些特定的目的将为一种新的血管内皮生长因子介导的基因传递方法提供体内的“原理证明”。优化的基于血管内皮生长因子的DNA载体和融合蛋白构建物将成为参与受体介导基因治疗的工业和学术团体的商业产品。拟议的商业应用：优化的基于血管内皮生长因子的DNA载体和用于刺激或抑制血管生成的融合蛋白的构建将成为参与受体介导的基因治疗的工业和学术团体的商业产品

The overall goal of this project is to develop a commercially viable system for selective gene delivery to growing endothelial cells. Growth of endothelial cells underlies angiogenesis that occurs almost exclusively in pathological situations and is promoted by the endothelial cell specific cytokine vascular endothelial growth factor (VEGF). In Phase l we proved the feasibility of a new methodology for VEGF- mediated gene delivery that eliminates direct chemical treatment of the growth factor or DNA. In Phase II studies we will construct transgenes for expression in endothelial cells and we will optimize a VEGF-based vector for highly selective DNA delivery to endothelial cells at angiogenesis sites in vivo. For stimulation of angiogenesis we propose to deliver a transgene encoding VEGF. For inhibition of angiogenesis we will construct transgenes for three apoptosis-inducing proteins fused to the herpesvirus protein vP22 that would ensure intercellular trafficking of the fusion proteins. The proteins we have chosen are: p53, procaspase-8a and granzyme B. Accomplishing these specific aims will provide the in vivo "proof-of- principle" for a novel methodology for VEGF-mediated gene delivery. The optimized VEGF-based DNA vehicle and constructs for fusion proteins will be commercial products for industrial and academic groups involved in receptor-mediated gene therapy. PROPOSED COMMERCIAL APPLICATIONS: The optimized VEGF-based DNA vehicle and constructs for fusion proteins that stimulates or inhibit angiogenesis will be commercial products for industrial and academic groups involved in receptor- mediated gene therapy