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: 8103513
• 负责人: Robert J Levy
• 金额: $ 25.13万
• 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/8103513
• 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. PUBLIC HEALTH RELEVANCE: The use of stents for treating vaso-occlusive disease has 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 in coronary disease (not peripheral vascular disease), but with significant risks. This proposal will investigate magnetic cell targeting of endothelial progenitor cells to stented arteries as a novel approach for regenerating diseased blood vessels.

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。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内皮祖细胞（EPC）产生磁性响应，从而能够将具有磁性靶向的治疗相关转基因传递到展开的支架，从而增强受损和患病动脉的再生。目标 1：制定并表征磁性靶向复合物的成分，包括：可生物降解的非聚合 MNP、EPC 和辅助依赖性腺病毒载体 (HD-Ad)。这些基因传递实验中使用的 MNP 可以通过在其成分中包含磁铁矿来实现磁响应，而不会影响其生物相容性。将从外周血中收获 EPC，并在 MNP 负载条件下研究磁响应性和活力。将与贝勒医学院的 Phillip Ng 博士合作构建 HD-Ad 载体，无需基因组整合即可实现持续转基因表达，并使用两种报告基因构建体（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。目标 2：研究带有报告基因构建体的磁性细胞体内靶向，以确定最佳的递送条件范围。将在大鼠颈动脉支架手术中研究预载有 MNP 的 GFPHD-Ad 转导 EPC，比较针对 304 钢支架的 EPC 与对照，以确定最佳递送条件以及表达 GFP 的 EPC 在目标动脉段中的分布。长达 28 天的长期研究将跟踪 LucHD-Ad 转导的 EPC 靶向的 304 个支架中 Luc 的表达，并通过 Luc 光学成像跟踪转基因表达随时间的变化。目标 3：研究 iNOSHD-Ad 转导的 MNP 修饰的 EPC 的体内抗再狭窄功效。根据 Aim 2 结果建立了最佳细胞制备和递送方案后，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再狭窄效果。 公众健康相关性：使用支架治疗血管闭塞性疾病与治疗效果和显着并发症有关，包括支架置入后的再梗阻，称为支架内再狭窄。药物洗脱支架已被证明可以改善冠心病（而非外周血管疾病）的支架内再狭窄，但风险也很大。该提案将研究磁性细胞将内皮祖细胞靶向支架动脉，作为再生患病血管的新方法。