Gene Delivery Stents

描述（由申请人提供）： 可扩张金属支架已成功用于缓解冠状动脉阻塞。在过去的一年里，美国使用了超过五十万个冠状动脉支架。此外，人们对预防支架血管成形术后再狭窄的药物输送支架越来越感兴趣。我们的实验室率先使用支架作为动脉壁基因治疗的基因传递系统平台。在本研究计划中，我们将提出以下假设：通过基因递送支架可以实现支架内再狭窄和易损斑块稳定的基因治疗。我们的基因递送支架利用抗体介导的复制缺陷腺病毒基因载体的束缚；这导致增强的位点特异性 ta'ansgene 表达，以及仅限于支架部署位点的高度局部化的生物分布。目标 目标 1：聚氨基二膦酸盐-钢相互作用以及随后的聚胺和抗体结合：配方和表征。双膦酸盐化学吸附将是直接或用放大多胺对钢支架进行分子表面修饰的基础，以允许使用双功能交联将抗腺病毒抗体与氨基共价结合，从而实现载体束缚。目标 2：钢-PAABP-抗体基因传递系统的配制和表征：细胞培养研究。动脉平滑肌细胞培养物将用作模型系统来研究基因传递机制，fi-半乳糖苷酶（LacZ）和绿色荧光蛋白（GFP）将用作这些实验的报告基因。目标3：体内效率和抗支架内再狭窄功效。猪冠状动脉支架研究将使用基于目标 1 和 2 的最佳配方。记者研究 (LacZ) 将重点关注载体的体内效率和生物分布。治疗基因将是 Fas-Ligand，一种促凋亡蛋白，对再狭窄具有确定的功效。主要治疗终点是新内膜形成的抑制程度。

DESCRIPTION (provided by applicant): Expandable metallic stents have been successfully utilized to relieve coronary arterial obstruction During this past year in the United States more then five hundred thousand coronary stents were used. Furthermore, there has been increasing interest in drug delivery stents to prevent restenosis following stent angioplasty. Our laboratory has pioneered the use of stents as platforms for gene delivery systems for arterial wall gene therapy. In this research program, we will address the following hypothesis: Gene therapy for in-stent restenosis and stabilization of vulnerable plaque can be achieved with a gene delivery stent. Our gene delivery stent utilizes antibody-mediated tethering of replication defective adenoviral gene vectors; this results in enhanced site specific ta'ansgene expression, and a highly localized biodistribution restricted to the site of stent deployment. Aims Aim 1: Polyaminobisphosphonate-steel interactions with subsequent polyamine and antibody binding: Formulation and Characterization. Bisphosphonate chemosorption will be the basis for a molecular surface modification of the steel stents either directly or with amplifying polyamines, to permit the covalent binding of anti-adenoviral antibodies to amino groups using bifunctional crosslinking, thereby enabling vector tethering. Aim 2: Formulation and characterization of the steel-PAABP-antibody gene delivery system: Ceil culture studies. Arterial smooth muscle cell cultures will be used as a model system to investigate the mechanism of gene delivery, fi-galactosidase (LacZ) and Green Fluorescent Protein (GFP) will be used as the reporter genes for these experiments. Aim 3: In vivo efficiency and anti-in-stent restenosis efficacy. Pig coronary stent studies will use the optimal formulations based on Aims 1 and 2. Reporter studies (LacZ) will focus on in vivo efficiency and biodistribution of vector. The therapeutic gene will be Fas-Ligand, a pro-apoptotic protein with established efficacy for restenosis. The chief therapeutic endpoint will be extent of inhibition of neointimal formation.