Gene Delivery in Retinal Diseases

描述(申请人提供)：将基因和药物输送到视网膜细胞是治疗视网膜和其他疾病的一个重大挑战。我们已经开发了一种非侵入性的方法，利用跨巩膜施加的电场来有效地将更多的RNA或DNA输送到视网膜色素上皮(RPE)。这种非侵入性的方法将避免手术视网膜下病毒输送可能产生的副作用，而且重复给药应该容易和实用。在未来，一旦在目前简单的模型系统中开发，这些结果将指导我们将基因疗法应用于眼科实践，以治疗包括青光眼和视网膜和黄斑变性在内的疾病。 这项研究选择的疾病和基因缺陷是已知可以在动物模型中治疗的，先前的工作作为指导和基线，与我们的交付方法进行疗效比较。 我们的DNA递送系统已经建立，并已满足经巩膜外DNA递送的优先功能。电场将被改进，以帮助和建立体内研究的启动参数。计划进行安全性研究，以帮助在不损害眼睛组织或动物的情况下实现高质量的交付。在优化实验的基础上进行迭代改进，并收集体外和体内的结果指标。在研究的后期阶段，获得了疗效数据。 我们计划在这个项目中学习何时以及如何预防由RPE65突变引起的LCA严重夜盲和视网膜变性。这项研究项目有两个目标。第一个目标是优化可以跨巩膜进入RPE的核酸的数量。第二个目标是评估RPE65基因在体内传递和表达的安全性和有效性。

DESCRIPTION (provided by applicant): Delivery of genes and drugs to the cells of the retina is a significant challenge in the treatment of retinal and other disorders. We have developed a non-invasive method that exploits electrical fields applied trans- sclerally to efficiently deliver larger amounts of RNA or DNA to the retinal pigmented epithelium (RPE). This noninvasive approach will avoid side effects that can occur with surgical subretinal viral delivery, and repeated dosing should be easy and practical. In the future, once developed in the present simple model system, these results will guide us in adapting gene therapy to ophthalmic practice to treat diseases including glaucoma and retinal and macular degenerations. The disease and gene defect selected for this study is known to be treatable in animal models, and prior work serves as guidance and as a baseline for efficacy comparisons with our delivery approach. Our DNA delivery systems are established and meet the priority function of trans-scleral DNA delivery in vitro already. The electrical fields will be refined to aid and establish starting parameters for in vivo studies. Safety studies are planned to help achieve high delivery without damage to eye tissues or the animal. Iterative improvements are made based on optimization experiments, and outcome measures are collected both in vitro and in vivo. In the latter stages of the study, efficacy data are obtained. We plan to learn when and how to prevent severe night blindness and retinal degeneration in LCA caused by mutations in RPE65 in this project. This research project has two goals. The first goal is to optimize the amounts of nucleic acids that can be driven into the RPE trans-sclerally. The second goal is to evaluate the safety and efficacy of RPE65 gene delivery and expression in vivo in mice with lesions in this gene.