Gene Delivery in Retinal Diseases

视网膜疾病中的基因传递

基本信息

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
项目类别：
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

项目摘要

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）：将治疗构建体传递到后眼是治疗视网膜和视网膜色素上皮（RPE）的盲目疾病的持续问题。为了应对这一挑战，我们的长期目标是在基因输送到RPE中的细胞外和细胞内贩运方面的见解。在过去的赠款期间，我们证明了用质粒DNA的眼部电穿孔后，小鼠RPE中报告基因的体内表达较高。最近，由于其特定的尺寸，核心和外壳设计，我们开发了新的纳米颗粒，其功能优势比前身具有许多优势。在初步数据中，我们表征了新颗粒的药代动力学。我们表明它们到达后段并有效地转染视网膜细胞，使其非常适合实用基因治疗。中央假设 - 透明质酸涂层的纳米颗粒（HA-NP）可以提供其他非病毒方法不能：1。它们通过自然和粘合性穿过间质空间。 2。它们通过脂质摄取可以有效地穿透细胞膜。 3。借助抑制剂，避免了细胞内和胞质外源-DNA宿主先天防御。 4。它们可以有效地运输到微管上的细胞核上。我们的研究将显示HA-NP如何克服这些非病毒基因疗法的经典障碍：我们将学习这四个关键特征的机制。这些HA-NP（以及他们的传递机理的了解以及如何操纵它们以进行优化的递送）可能在其他非病毒基因治疗方法可能没有的基因治疗中有效。对现场的影响：这项研究应改善基因递送，以至于患者可以用眼药水自我治疗。使用局部眼部滴剂将基因治疗剂传递到后段，而RPE具有变化性。