Gene Delivery in Retinal Diseases

视网膜疾病中的基因传递

• 批准号: 8689037
• 负责人: John M Nickerson
• 金额: $ 36.97万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8689037
• 关键词: Adhesiveness; Algorithms; Anterior; Applications Grants; Avidity; Back; Binding; Biodistribution; Biological Preservation; Blindness; Blood Circulation; Caliber; Cell Culture Techniques; Cell Nucleus; Cell membrane; Cell surface; Cells; Clathrin; Collaborations; Cultured Cells; Cytoplasm; DNA; DNA Sequence; Data; Disease; Dose; Drug Delivery Systems; Drug Formulations; Drug Kinetics; Electroporation; Electroretinography; Enhancers; Evaluation; Extracellular Matrix; Eye; Eyedrops; Figs - dietary; Gatekeeping; Gene Delivery; Gene Expression; Gene Expression Profiling; Gene Therapy Agent; Genes; Goals; Grant; Hyaluronan; Hyaluronic Acid; Immune response; Knowledge; Lead; Learning; Life; Measures; Mediating; Membrane Microdomains; Microscopy; Microtubules; Monitor; Morphology; Movement; Mus; Nuclear; Particle Size; Patients; Penetration; Pharmacologic Substance; Polysaccharides; Public Health; Reporter Genes; Research; Retina; Retinal; Retinal Diseases; Route; Science; Sclera; Structure of retinal pigment epithelium; Surface; System; Testing; Therapeutic; Time; Topical application; Treatment Protocols; Viral; Vision; base; design; extracellular; gene therapy; improved; in vivo; in vivo Model; inherited retinal degeneration; inhibitor/antagonist; insight; interstitial; meetings; mouse model; nanoparticle; non-viral gene therapy; novel; particle; plasmid DNA; prevent; promoter; research study; small molecule; success; therapeutic DNA; therapeutic gene; trafficking; uptake

DESCRIPTION (provided by applicant): Delivery of therapeutic constructs to the posterior eye is an ongoing problem in treating blinding diseases of the retina and retinal pigmented epithelium (RPE). To meet this challenge, our long- term goal is to gain insights into extra- and intracellular trafficking in gene delivery to the RPE. In the past grant period, we demonstrated high in vivo expression of reporter genes in mouse RPE following ocular electroporation with plasmid DNA. Recently, we developed new nanoparticles that have many functional advantages over predecessors due to their specific sizing, core, and shell design. In our preliminary data, we characterized the pharmacokinetics of our new particles. We showed that they reach the posterior segment and transfect retinal cells efficiently, making them ideal for practical gene therapy. Central hypotheses-Hyaluronan coated nanoparticles (HA-NPs) can deliver cargos that other nonviral approaches cannot: 1. They penetrate through interstitial spaces through avidity and adhesiveness. 2. They penetrate cell membranes efficiently through lipid-raft uptake. 3. With aid of inhibitors, they avoid intracellular and cytosolic foreign-DNA host innate defenses. 4. They can be efficiently transported to the nucleus on microtubules. Our studies will show how the HA-NPs overcome these classic barriers to nonviral gene therapy: We will learn the mechanisms of these four key features. These HA-NPs (and knowledge of their mechanism of delivery, and how to manipulate them for optimized delivery) are likely to be effective in gene therapy where other nonviral gene therapy approaches might not. Impact on Field: This research should improve gene delivery to the point that a patient can self- treat with eye drops. The use of topical eye drops to deliver gene therapy agents to the posterior segment and the RPE is transformative.

描述（由申请人提供）：将治疗结构递送至后眼是治疗视网膜和视网膜色素上皮（RPE）致盲疾病中持续存在的问题。为了应对这一挑战，我们的长期目标是深入了解 RPE 基因传递过程中的细胞外和细胞内运输。在过去的资助期内，我们在用质粒 DNA 进行眼部电穿孔后，证明了小鼠 RPE 中报告基因的高体内表达。最近，我们开发了新的纳米颗粒，由于其特定的尺寸、核和壳设计，与前代产品相比具有许多功能优势。在我们的初步数据中，我们描述了新颗粒的药代动力学。我们证明它们能够到达后段并有效转染视网膜细胞，这使它们成为实际基因治疗的理想选择。中心假设 - 透明质酸包被的纳米颗粒 (HA-NP) 可以传递其他非病毒方法无法传递的货物： 1. 它们通过亲和力和粘附性穿透细胞间隙。 2. 它们通过脂筏摄取有效地穿透细胞膜。 3. 在抑制剂的帮助下，它们可以避免细胞内和胞质外源DNA宿主的先天防御。 4. 它们可以通过微管有效地转运到细胞核。我们的研究将展示 HA-NP 如何克服非病毒基因治疗的这些经典障碍：我们将了解这四个关键特征的机制。这些 HA-NP（以及对其传递机制的了解，以及如何操纵它们以优化传递）可能在基因治疗中有效，而其他非病毒基因治疗方法可能无效。对领域的影响：这项研究应该改善基因传递，使患者可以用滴眼液进行自我治疗。使用局部滴眼剂将基因治疗药物输送至眼后段和 RPE 具有变革意义。