Gene Delivery Stents

描述(由申请人提供)：在过去的一年里，美国已经成功地使用可膨胀金属支架来缓解冠状动脉阻塞，使用了超过50万个冠状动脉支架。此外，药物输送支架在预防支架血管成形术后再狭窄方面的应用也越来越受到关注。我们的实验室率先使用支架作为基因输送系统的平台，用于动脉壁基因治疗。在这个研究项目中，我们将解决以下假设：通过基因输送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。我们的基因输送支架利用抗体介导的复制缺陷型腺病毒基因载体的连接；这导致增强了部位特异性反式基因的表达，并且高度局限于支架部署部位的生物分布。目的：目的1：多氨基二膦酸盐-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成为直接或与放大多胺一起对钢支架进行分子表面修饰的基础，以允许抗腺病毒抗体通过双功能交联剂与氨基共价结合，从而实现载体拴系。目的2：STEAR-PAABP-抗体基因递送系统的研制及特性：细胞培养研究。本实验将以动脉平滑肌细胞培养为模型系统，以半乳糖苷酶(LacZ)和绿色荧光蛋白(GFP)为报告基因，研究基因传递机制。目的3：体内疗效和抗支架内再狭窄的疗效。猪冠状动脉支架的研究将使用基于AIMS 1和2的最佳配方。报告研究(LacZ)将重点放在载体的体内效率和生物分布上。治疗基因将是Fas-Ligand，一种对再狭窄已有疗效的促凋亡蛋白。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.