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）：我们将研究一种提高支架血管成形术治疗效果的新方法。假设可生物降解的非聚合物磁性纳米颗粒（MNP）可用于使离体培养的循环内皮祖细胞（EPC）具有磁响应性，从而能够将具有磁性靶向的治疗相关转基因递送至展开的支架，从而能够增强损伤和患病动脉的再生。目标1：配制和表征磁性靶向复合物的组分，包括：可生物降解的非聚合物MNP、EPC和辅助依赖性腺病毒载体（HD-Ad）。在这些基因递送实验中使用的MNP可以通过在其组合物中包含磁铁矿而呈现磁响应性，而不会损害其生物相容性。将从外周血收获EPC，并在MNP负载条件下研究其磁响应性和活力。将与贝勒医学院的菲利普·吴博士合作构建HD-Ad载体，其导致持续的转基因表达而没有基因组整合，其中两种报告构建体（绿色荧光蛋白，GFP和荧光素酶，Luc）以及诱导型一氧化氮合酶（iNOS）作为我们的主要治疗基因。目的2：研究磁性报告基因在体内的靶向性，建立最佳的递送条件范围。将在大鼠颈动脉支架手术中研究预装载有MNP的GFPHD-Ad转导的EPC，比较靶向304钢支架的EPC与对照，以确定最佳递送条件和表达GFP的EPC在靶向动脉段中的分布。更长期的研究，长达28天，将在304个用LucHD-Ad转导的EPC靶向的支架中跟踪Luc表达，随后用Luc光学成像随时间推移进行转基因表达。目的3：研究iNOSHD-Ad转导的MNP修饰的EPC在体内抗再狭窄的作用。根据Aim 2结果建立了最佳细胞制备和递送的方案，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动脉支架血管成形术中的抗再狭窄作用。