High Field Gradient Targeting Magnetic Nanoparticle Loaded Cells to Stents

高场梯度将磁性纳米粒子负载细胞靶向支架

• 批准号: 8270523
• 负责人: Robert J Levy
• 金额: $ 20.53万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8270523
• 关键词: Adenovirus Vector; Affect; Angioplasty; Aortic Aneurysm; Arteries; Autologous; Blood Vessels; Carotid stent; Cells; Clinical Trials Design; Collaborations; Complex; Control Groups; Coronary; Coronary Arteriosclerosis; Coronary heart disease; Drug usage; Endothelial Cells; Family suidae; Feasibility Studies; Gene Delivery; Generations; Genomics; Green Fluorescent Proteins; Harvest; Lead; Longitudinal Studies; Luciferases; Magnetism; Mediating; Medicine; Metals; Modeling; Natural regeneration; Outcome; Paper; Peripheral Vascular Diseases; Peripheral arterial disease; Pharmaceutical Preparations; Procedures; Protocols documentation; Publishing; Rattus; Reporter; Risk; Steel; Stem cells; Stents; Study of magnetics; Therapeutic; Time; Transgenes; Treatment Efficacy; Vascular Graft; Veno-Occlusive Disease; base; biomaterial compatibility; cell preparation; college; effective therapy; femoral artery; helper-dependent adenoviral vector; human NOS2A protein; improved; in vivo; injured; interest; magnetite ferrosoferric oxide; nanoparticle; non-viral gene delivery; novel strategies; optical imaging; peripheral blood; research study; restenosis; standard of care; therapeutic gene; transgene expression; vector; viral gene delivery

DESCRIPTION (provided by applicant): We will investigate a novel approach to improve the therapeutic efficacy of stent angioplasty. It is hypothesized that biodegradable nonpolymeric magnetic nanoparticles (MNP) can be used to render ex vivo cultured circulating endothelial progenitor cells (EPC) magnetically responsive and thus able to deliver a therapeutically relevant transgene with magnetic targeting to a deployed stent thereby enabling enhanced regeneration of injured and diseased arteries. Aim 1: To formulate and characterize the components of a magnetic targeting complex including: Biodegradable nonpolymeric MNP, EPC and Helper Dependent Adenoviral Vectors (HD-Ad). MNP used in these gene delivery experiments can be rendered magnetically responsive by inclusion of magnetite in their composition without compromising their biocompatibility. EPC will be harvested from peripheral blood and investigated under conditions of MNP loading for magnetic responsiveness and viability. HD-Ad vectors, that result in sustained transgene expression without genomic integration will be constructed in collaboration with Dr. Phillip Ng, Baylor College of Medicine, with both reporter constructs (green fluorescent protein, GFP, and luciferase, Luc), as well as inducible nitric oxide synthase (iNOS) as our lead therapeutic gene. Aim 2: To study magnetic cell targeting in vivo with reporter constructs to establish an optimal range of delivery conditions. GFPHD-Ad transduced EPC preloaded with MNP will be investigated in rat carotid stent procedures comparing EPC targeted to 304 steel stents versus controls to determine optimal delivery conditions and the distribution of the GFP expressing EPC in the targeted arterial segment. Longer term studies, up to 28 days, will follow Luc expression in 304 stents targeted with EPC transduced with LucHD-Ad, following transgene expression over time with Luc optical imaging. Aim 3: To investigate in vivo the antirestenotic efficacy of MNP-modified EPC transduced with iNOSHD-Ad. Having established protocols for optimal cell preparation and delivery per Aim 2 results, we will investigate the antirestenotic effect of magnetically targeted EPC modified with iNOSHD-Ad in both the rat carotid stenting model and pig femoral stent angioplasties.

描述（由申请人提供）：我们将研究一种新的方法来提高支架血管成形术的治疗功效。据推测，可生物降解的非聚合磁性纳米颗粒（MNP）可用于使离体培养的循环循环内皮祖细胞（EPC）具有磁响应性，从而能够将磁性靶向与磁性靶向进行治疗相关的靶向，从而向其部署的稳固型固定构成，使其受益于增强的调节性，并具有不良的调节性。 AIM 1：制定和表征磁靶向复合物的成分，包括：可生物降解的非聚合物MNP，EPC和辅助腺病毒载体（HD-AD）。这些基因递送实验中使用的MNP可以通过将磁铁矿包含在其组成中而不会损害其生物相容性来对磁响应。 EPC将从外周血中收获，并在MNP加载条件下进行磁反应性和生存能力进行研究。 HD-AD矢量，导致不持续的转基因表达而无需基因组整合，将与Phillip Ng博士合作构建Baylor Medicine College，均具有记者构造（绿色荧光蛋白，GFP和Luciferase，Luc），以及可诱导的一氧化物合酶（Inos）以及我们的领先基因。 AIM 2：使用报告构建体的体内靶向磁细胞靶向，以建立最佳的输送条件范围。将研究用MNP预加载的GFPHD AD转导的EPC在大鼠颈动脉支架程序中进行研究，将靶向的EPC与304钢支架与对照组相比，以确定最佳的递送条件和在靶向动脉段中表达EPC的GFP分布。长期研究（长达28天）将跟随304个支架的LUC表达，该支架用LuCHD-AD转导的EPC靶向，随着时间的流逝，LUC光学成像随着时间的流逝。目的3：在体内研究MNP修饰的EPC的抗雷神疗效。在建立了以最佳细胞制备和每个AIM 2的结果为结果的方案，我们将研究用InoSHD-AD修饰的磁靶向EPC在大鼠颈动脉支架模型和猪股骨支架血管成形术中的抗雷神效应。