NEW METHODOLOGY FOR VEGF-MEDIATED GENE DELIVERY

VEGF 介导的基因传递的新方法

• 批准号: 6072495
• 负责人: Joseph M Backer
• 金额: $ 36.97万
• 依托单位: SIBTECH, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6072495
• 关键词: angiogenesis; angiogenesis inhibitors; biotechnology; cell line; chimeric proteins; cysteine endopeptidases; gene expression; gene targeting; gene therapy; laboratory mouse; method development; p53 gene /protein; protein engineering; protein purification; protein tyrosine kinase; serine proteinases; transfection; transfection /expression vector; vascular endothelial growth factors; vascular endothelium; virus protein

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

该项目的总体目标是开发一种商业上可行的系统，用于选择性基因递送到生长的内皮细胞。内皮细胞的生长是血管生成的基础，血管生成几乎只发生在病理情况下，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I期，我们证明了VEGF介导的基因递送新方法的可行性，该方法消除了生长因子或DNA的直接化学处理。在第二阶段研究中，我们将构建用于在内皮细胞中表达的转基因，并优化基于VEGF的载体，以高选择性地将DNA递送到体内血管生成部位的内皮细胞。为了刺激血管生成，我们提出递送编码VEGF的转基因。为了抑制血管生成，我们将构建三种与疱疹病毒蛋白vP 22融合的促增殖蛋白的转基因，这将确保融合蛋白的细胞间运输。我们选择的蛋白质是：p53，procaspase-8a和颗粒酶B。实现这些特定的目标将提供一种新的方法VEGF介导的基因传递在体内的“原理证明”。优化的基于VEGF的DNA载体和融合蛋白的构建体将成为工业和学术团体参与受体介导的基因治疗的商业产品。拟议的商业应用：优化的基于VEGF的DNA载体和用于刺激或抑制血管生成的融合蛋白的构建体将成为涉及受体介导的基因治疗的工业和学术团体的商业产品