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。在过去的一年中，美国使用了超过五十万个冠状动脉支架。此外，人们对药物递送支架以防止支架血管成形术后再狭窄的兴趣越来越大。我们的实验室率先使用支架作为动脉壁基因治疗的基因递送系统平台。在这项研究计划中，我们将解决以下假设：基因治疗支架内再狭窄和稳定的易损斑块可以实现与基因输送支架。我们的基因递送支架利用复制缺陷型腺病毒基因载体的抗体介导的拴系;这导致增强的位点特异性taans基因表达，以及限制于支架部署位点的高度局部化的生物分布。目的目的1：聚氨基二膦酸-钢与随后的聚胺和抗体结合的相互作用：制剂和表征。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改性的基础，以允许使用双官能交联将抗腺病毒抗体共价结合至氨基，从而实现载体拴系。目的2：钢-PAABP-抗体基因递送系统的配制和表征：细胞培养研究。动脉平滑肌细胞培养物将用作研究基因递送机制的模型系统，β-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。目的3：体内有效性和抗支架内再狭窄有效性。猪冠状动脉支架研究将使用基于目标1和2的最佳配方。报告基因研究（LacZ）将侧重于载体的体内效率和生物分布。治疗基因将是Fas-配体，一种对再狭窄具有确定疗效的促凋亡蛋白。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.