Gene Delivery Stents

基因递送支架

• 批准号: 8051784
• 负责人: Robert J Levy
• 金额: $ 40.31万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8051784
• 关键词: Address; Adenoviruses; Amplifiers; Angioplasty; Animals; Antineoplastic Agents; Binding; Binding Sites; Blood Vessels; Cell Culture Techniques; Chemicals; Chemistry; Complex; Coronary Arteriosclerosis; Coronary heart disease; Cytomegalovirus; Dose; Dose-Limiting; Drug Formulations; Feasibility Studies; Gene Delivery; Generations; Genes; Goals; Green Fluorescent Proteins; Health; Imines; Immune; Immunosuppressive Agents; In Vitro; Inflammatory; Investigation; Lead; Learning; Mediating; Medicine; Metals; Modeling; Molecular Weight; Natural regeneration; Outcome; Paclitaxel; Patients; Pharmaceutical Preparations; Polyamines; Polyethylenes; Polymers; Publications; Rattus; Reporter; Reporting; Research; Risk; Sirolimus; Site; Smooth Muscle Myocytes; Steel; Stents; Study Section; Sulfhydryl Compounds; Surface; Therapeutic; Therapeutic Effect; Therapeutic Studies; Time; Transgenes; Treatment Efficacy; Vascular Diseases; Water; base; bisphosphonate; college; dosage; gene therapy; helper-dependent adenoviral vector; human NOS2A protein; improved; in vivo; innovation; novel; novel strategies; optical imaging; polyallylamine; prevent; promoter; response; restenosis; synthetic construct; therapeutic gene; therapeutic transgene; time use; transduction efficiency; transgene expression; vector

DESCRIPTION (provided by applicant): Stent angioplasty over the last decade has made a major impact on improved therapeutic outcomes for coronary disease. Our group has pioneered the concept that an even greater therapeutic benefit than possible with current bare metal stents (BMS) or drug eluting stents (DES) could be achieved with stent-mediated gene therapy from the bare metal surfaces of stents. This proposal will address the following questions: 1) Can a high enough dose of a gene vector be loaded onto a stent? 2) Can transgene expression be sustained long enough to achieve a therapeutic benefit? The first question is now addressed through our investigations of a three component synthetic complex that includes a synthetic amplifier construct (AC) thereby enabling higher dosing. Prolonged expression will be addressed through our use of Helper Dependent Adenoviruses (HD-Ad) that can demonstrate transgene expression in vivo for several years. Aim 1. To investigate the syntheses of novel Amplifier Constructs (AC) for attaching higher levels of HD-Ad to the bare metal surfaces of stents. The overall complex consists of: 1) A water soluble bisphosphonate (PABT) with irreversible binding to stent surfaces; 2) A hydrolysable linker (HL) that can be reacted with Ad (via sulfyhydryl chemistry) for tethering Ad to stents; and 3) An amplifier construct (AC). This Aim will focus on novel AC synthetic constructs to increase HD-Ad doses. Aim 2. To investigate and model in cell culture, local delivery of HD-Ad from steel surfaces, in order to assess the site specific efficiency and therapeutic potential of novel synthetic ACs. We will create HD- Ad vectors: For reporter HD-Ad cell culture studies, we will create a green fluorescent protein (GFP) vector under the control of the cytomegalovirus promoter (CMV). We will also create a HD-Ad luciferease (LUC) construct with a CMV promoter to be evaluated in vitro and also used in the Aim 3 in vivo studies for optical imaging studies. Our therapeutic vector will include rat inducible nitric oxide synthase (iNOS) with the rat arterial smooth muscle cell specific SM22alpha promoter, to both target smooth muscle cells and to minimize the effects of immune factors on both the efficacy and duration of therapeutic transgene activity. Aim 3. To investigate in vivo the efficiency and efficacy of AC enhanced local delivery of HD-Ad from stent surfaces. These investigations will study the dose ranging and therapeutic dose response capabilities of our three component complex approach to gene delivery from stent surfaces. Reporter studies delivering LUCHD-Ad will investigate transgene activity over time using in vivo LUC optical imaging of stent delivery. Once the lead AC formulation with the greatest in vivo delivery capacity of reporter HD-Ad (re. the upper limits of the dose range) has been characterized, therapeutic studies will be carried out investigating sustained transgene expression and anti-instent restenosis efficacy over time with iNOSHD-Ad delivery using our 3 component complex with maximal AC HD-Ad delivery capacity. PUBLIC HEALTH RELEVANCE The use of stents for treating coronary artery disease had been associated with both therapeutic benefit and significant complications, including reobstruction post-stenting, termed instent restenosis. Drug eluting stents have been shown to ameliorate instent restenosis, but with significant risks. This proposal will investigate gene delivery stents as a novel approach for regenerating diseased blood vessels.

描述（由申请人提供）：过去十年中支架血管成形术对改善冠状动脉疾病的治疗结果产生了重大影响。我们的小组率先提出了这样的概念，即使用裸架的裸型金属表面可以实现当前裸金属支架（BMS）或药物洗脱支架（DES）的更大的治疗益处。该建议将解决以下问题：1）可以将足够高的基因载体加载到支架上吗？ 2）转基因表达能够维持足够长的时间以获得治疗益处吗？现在，通过我们对包括合成放大器构建体（AC）的三个成分合成复合物的研究来解决第一个问题，从而实现了更高的剂量。长时间的表达将通过我们使用辅助腺病毒（HD-AD）来解决，这些腺病毒（HD-AD）可以证明体内的转基因表达数年。目的1。研究新型放大器构建体（AC）的合成，以将更高水平的HD-AD连接到支架的裸金属表面上。总体复合物由：1）水溶性双膦酸盐（PABT）具有不可逆转的结合与支架表面的结合； 2）可以与AD（通过亚硫赖尔化学）反应的水解接头（HL）将AD绑定到支架上； 3）放大器构建体（AC）。该目标将集中在新型的AC合成构建体上，以增加HD-AD剂量。目的2。要研究和模拟细胞培养物，即从钢表面局部输送HD-AD，以评估新型合成ACS的特定效率和治疗潜力。我们将创建HD-AD向量：对于报告基因AD细胞培养研究，我们将在巨细胞病毒启动子（CMV）控制下创建绿色荧光蛋白（GFP）载体。我们还将使用CMV启动子在体外评估的CMV启动子创建HD-AD LUCIFERESE（LUC）构建体，并在AIM 3体内研究中用于光学成像研究。我们的治疗载体将与大鼠动脉平滑肌细胞特异性SM22Alpha启动子一起包括大鼠诱导的一氧化氮合酶（INOS），以靶向平滑肌细胞，并最大程度地降低免疫因子对治疗转基因活性疗效和持续时间的影响。目的3。研究体内AC从支架表面增强HD-AD的局部输送的效率和功效。这些研究将研究我们三个成分复杂方法从支架表面递送基因的剂量范围和治疗剂量反应能力。提供Luchd-AD的记者研究将使用支架递送的体内LUC光学成像随着时间的推移研究转基因活性。一旦将记者HD-AD的体内输送能力最大的铅AC配方（剂量范围的上限）进行了表征，则将进行治疗研究，以调查持续的转基因表达和抗稳定的再肾上腺增生功效，随着时间的推移，使用INOSHD-AD递送，使用我们的3个组件，使用我们的3个组件复合物与最大值AC HD-AC HD-AC HD-AC HD-AC HD-AC HD-AC HD-AC HD-AC HD-AC HD-AC HD-AC HD-AD交付。公共卫生相关性使用支架治疗冠状动脉疾病与治疗益处和显着并发症有关，包括后结构后的重新粘结，称为锻炼。药物洗脱支架已被证明可以改善体内再狭窄，但具有很大的风险。该提案将研究基因递送作为再生患病血管的新方法。