Gene Delivery in Retinal Diseases

视网膜疾病中的基因传递

• 批准号: 8519457
• 负责人: John M Nickerson
• 金额: $ 35.84万
• 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/8519457
• 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 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）：将治疗性构建体递送至后眼是治疗视网膜和视网膜色素上皮（RPE）致盲性疾病的一个持续存在的问题。为了迎接这一挑战，我们的长期目标是深入了解基因递送到RPE的细胞外和细胞内运输。在过去的资助期间，我们证明了在小鼠RPE中报告基因的高体内表达与质粒DNA眼电穿孔。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，与前辈相比具有许多功能优势。在我们的初步数据中，我们描述了我们的新颗粒的药代动力学。我们发现，它们可以有效地到达后段并切除视网膜细胞，使其成为实用基因治疗的理想选择。中心假设--它们通过嗜酸性和嗜酸性穿透细胞间隙。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，它们避免细胞内和胞质外源DNA宿主的先天防御。4. 它们可以在微管上被有效地运输到细胞核。我们的研究将展示HA-NPs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（及其传递机制的知识，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到病人可以用眼药水自我治疗的程度。使用局部滴眼液将基因治疗剂递送至眼后段和RPE是变革性的。