Gene Delivery Stents

描述（由申请人提供）：可膨胀金属支架已成功用于缓解冠状动脉阻塞在过去的一年里，在美国使用了超过50万个冠状动脉支架。此外，人们对药物输送支架越来越感兴趣，以防止支架成形术后再狭窄。我们的实验室率先使用支架作为动脉壁基因治疗基因传递系统的平台。在本研究计划中，我们将提出以下假设：基因传递支架可以实现支架内再狭窄的基因治疗和易损斑块的稳定。我们的基因传递支架利用抗体介导的复制缺陷腺病毒基因载体的系缚；这导致增强的位点特异性ta’ansgene表达，以及高度局限于支架部署部位的生物分布。目的1：聚氨基二磷酸盐-钢与随后的多胺和抗体结合的相互作用：配方和表征。双膦酸盐化学吸附将是钢支架分子表面修饰的基础，可以直接或与扩增的多胺一起修饰，以允许使用双功能交联将抗腺病毒抗体与氨基共价结合，从而实现载体捆绑。目的2：钢- paabp抗体基因传递体系的构建与表征：Ceil培养研究。动脉平滑肌细胞培养将作为研究基因传递机制的模型系统，fi-半乳糖苷酶（LacZ）和绿色荧光蛋白（GFP）将作为这些实验的报告基因。目的3：体内效率和抗支架内再狭窄疗效。猪冠状动脉支架研究将使用基于目标1和目标2的最佳配方。报告研究（LacZ）将重点关注媒介的体内效率和生物分布。治疗基因将是fas -配体，一种具有再狭窄疗效的促凋亡蛋白。主要的治疗终点将是抑制新生内膜形成的程度。

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.