Theranostics of Photoreceptor-RPE-Choroid Neurovascular Unit in Mouse Models of Eye Diseases

标题：眼病小鼠模型中光感受器-RPE-脉络膜神经血管单位的治疗诊断学 PI：Robert J. Zawadzki，博士 总结 遗传性和年龄相关性黄斑变性（AMD）目前是造成严重视力损害的原因 在超过200万美国居民中，随着人口老龄化，预计到2040年患病率将翻一番。 变性发生在感光细胞、视网膜色素上皮（RPE）细胞和脉络膜中。 血管系统是后眼中紧密相互依赖的组织的复合体。该项目将在 在两种黄斑变性的动物模型中， 线粒体功能障碍模型和Doyne Honeycomb Retinal Dystrophy。这些模型概括了两个 遗传性和年龄相关性黄斑变性、RPE细胞变性和年龄- RPE和Bruch膜之间细胞外沉积物的相关增加，Bruch膜将RPE分离 从脉络膜毛细血管床。该项目将使用创新的，细胞级分辨率的活体成像 结合额外的功能测试，以表征与年龄相关的结构和功能变化， 光感受器-RPE-脉络膜神经血管单位（PRC-NVU）的细胞。这些研究将在 在具有遗传缺陷的小鼠群和野生型对照中纵向地观察。这些研究将测试 RPE-Bruch膜中的原发性缺陷引起继发性恶化的假设 光感受器和脉络膜毛细血管系统。这些研究包括光感受器结构的测量 和电活动，视杆视色素视紫红质的漂白和再生，Bruch's的映射 膜厚度、RPE细胞自发荧光、RPE细胞的氧化还原状态和脉络膜毛细血管 形态和流动。在研究结束时，将对来自每只小鼠的一只眼睛的脉络膜-RPE进行分析。 用高分辨率离体共聚焦显微镜成像;将来自每只小鼠的第二只眼睛的视网膜 通过在光或电子上进行的常规组织学和生物化学测量进行评价 显微镜，以允许验证体内发现。一种新的方法，用于将药物通过近端递送到RPE， 红外光可降解纳米颗粒将用于将治疗剂局部靶向到患病的RPE细胞， 空间分辨成像将用于确定治疗剂是否减缓和停止疾病 进展通过结合纵向，体内成像和光学纳米治疗，这些研究将奠定一个 为人类眼部疾病的局部靶向药物输送奠定了基础。

TITLE: Theranostics of Photoreceptor-RPE-Choroid Neurovascular Unit in Mouse Models of Eye Diseases PI: Robert J. Zawadzki, Ph.D. SUMMARY Inherited and age-related macular degeneration (AMD) are currently responsible for serious vision impairment in over 2 million US residents, with prevalence expected to double by 2040 as the population ages. Degeneration occurs in photoreceptor cells, retinal pigment epithelial (RPE) cells and in the choroidal vasculature, a complex of tightly interdependent tissues in the posterior eye. This project will investigate in vivo the photoreceptor-RPE-choroid complex in animal models of two macular degenerations, RPE mitochondrial dysfunction model and Doyne Honeycomb Retinal Dystrophy. These models recapitulate two major hallmarks of inherited and age related macular degeneration, degeneration of RPE cells, and age- related increase in extracellular deposits between the RPE and Bruch's membrane, which separates the RPE from the choroidal capillary bed. The project will use innovative, cellular-level resolution in vivo imaging combined with additional functional tests to characterize age-related changes in the structure and function of cells of the photoreceptor-RPE-choroid neurovascular unit (PRC-NVU). These studies will be performed longitudinally in cohorts of mice with the genetic defects, and in wild type controls. The studies will test the hypothesis that the primary defects in RPE-Bruch's membrane cause secondary deterioration of photoreceptors and choriocapillaris vasculature. The studies include measures of photoreceptor structure and electrical activity, bleaching and regeneration of the rod visual pigment rhodopsin, mapping of Bruch's membrane thickness, RPE cell autofluorescence, the redox status of RPE cells, and choriocapillaris vascular morphology and flow. At the termination of the study, the choroid-RPE from one eye of each mouse will be imaged with high resolution ex vivo confocal microscopy; the retina from the second eye of each mouse will be evaluated by conventional histological and biochemical measurements performed on light or electron microscopes, to allow validation of in vivo findings. A novel method for delivering drugs to the RPE via near infrared light-degradable nanoparticles will be used to locally target therapeutic agents to ailing RPE cells, and spatially resolved imaging will be used to determine if the therapeutic agents slow and stop disease progression. By combining longitudinal, in vivo imaging and optical nanotherapies, these studies will lay a foundation for locally targeted drug delivery in human ocular disease.