Gene Delivery in Retinal Diseases

描述（由申请人提供）：将基因和药物递送到视网膜细胞是视网膜和其他疾病治疗中的一个重大挑战。我们开发了一种非侵入性方法，利用经巩膜施加的电场，有效地将大量RNA或DNA传递到视网膜色素上皮（RPE）。这种非侵入性方法将避免手术视网膜下病毒传递可能发生的副作用，并且重复给药应该是容易和实用的。在未来，一旦在目前简单的模型系统中发展，这些结果将指导我们将基因治疗应用于眼科实践，以治疗包括青光眼、视网膜和黄斑变性在内的疾病。

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.