Theranostics of Photoreceptor-RPE-Choroid Neurovascular Unit in Eye Diseases:

眼部疾病中光感受器-RPE-脉络膜神经血管单元的治疗诊断学：

基本信息

批准号：
10587983
负责人：
Robert J Zawadzki
金额：
$ 59.42万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-02-01 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10587983
关键词：
11 cis Retinal 3-Dimensional Acceleration Affect Age Age related macular degeneration Aging Angiography Animal Model Animals Basal lamina Biochemical Biological Assay Biological Markers Blood Circulation Blood Vessels Blood-Retinal Barrier Bruch&apos s basal membrane structure Cells Cellular Structures Cholesterol Choroid Complement Complex Custom Data Defect Degenerative Disorder Deposition Deterioration Diagnosis Diagnostic Diagnostic Procedure Dimensions Disease Disease Progression Doctor of Philosophy Doyne honeycomb retinal dystrophy Drug Delivery Systems Drug Targeting Electron Microscope Electroretinography Epithelial Cells Evaluation Exhibits Extracellular Matrix Eye Eye diseases Fluorescence Foundations Functional disorder Genetic Glucose Goals Grant Health High Fat Diet Histologic Hour Human Image Impairment Incubated Individual Inherited Investigation Label Light Light Microscope Link Lipids Macular degeneration Maps Measurement Measures Methods Microscope Microscopy Modeling Molecular Morphology Mus Mutation Natural regeneration Nutrient Opsin Optical Coherence Tomography Optical Methods Optics Organelles Oxidation-Reduction Oxygen Pathology Patients Pharmaceutical Preparations Photoreceptors Point Mutation Population Predictive Value Prevalence Recycling Resolution Retina Retinal Degeneration Retinal Photoreceptors Retinoids Role S1-5 protein Sclera Speed Stress Structure Structure of retinal pigment epithelium System Tail Testing Therapeutic Therapeutic Agents Thick Time Tissues Translations Validation Veins Visual impairment Vitamin A Water age related capillary bed cell type cohort confocal imaging density experimental study extracellular feeding imaging platform imaging system in vivo in vivo optical imaging innovation mitochondrial dysfunction mosaic multimodality nanoparticle nanotherapy neurovascular unit novel novel diagnostics ocular imaging optical spectra particle retinal imaging targeted treatment theranostics tool

项目摘要

TITLE: Theranostics of Photoreceptor-RPE-Choroid Neurovascular Unit in Eye Diseases PI: Robert J. Zawadzki, Ph.D. SUMMARY Inherited and age-related macular degeneration (AMD) are currently responsible for severe 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 the choroidal vasculature, a complex of tightly interdependent tissues in the posterior eye. This project will continue our in vivo investigations of morphology and function of the photoreceptor-RPE-choroid complex in animal models of retinal degenerations, including the RPE mitochondrial dysfunction model and Doyne Honeycomb Retinal Dystrophy. These models recapitulate 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 probing of retinal structure and function combined with additional functional tests to characterize age-related changes in major components of the photoreceptor-RPE-choroid neurovascular unit (PRC-NVU). These studies will be performed longitudinally in cohorts of mice with genetic defects and wild- type controls, fed with a high-fat diet (to accelerate disease progression). The studies will test the hypothesis that primary defects in RPE-Bruch’s membrane cause secondary deterioration of photoreceptors and choriocapillaris vasculature. The studies include measures of photoreceptor structure and function (using Temporal Speckle Averaging - Optical Coherence Tomography (TSA-OCT), OCT-based Optoretinography (ORG)), and full-field Electroretinography (ffERG)), mapping of RPE mosaic and Bruch’s membrane morphology with water driven transient changes in BrM scattering, RPE cell autofluorescence intensity and emission spectrum, the redox status of RPE cells, and choriocapillaris vascular morphology using TSA-OCT Angiography. We will analyze these multidimensional longitudinal data to discover potential structural and functional biomarkers of PRC-NVU degeneration and evaluate its predictive value for disease progression. At the termination of in vivo experiments, the choroid-RPE from one eye of each mouse will be imaged with a custom high-resolution ex vivo confocal microscope equipped with an Optical Coherence Microscopy (OCM) unit and a LiveCell stage top incubation system allowing imaging of a “live” RPE cells flat-mounted with the sclera (to allow validation of in vivo observations and evaluation of Fluorescence Lifetime of RPE organelles); 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 morphological 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.

标题：眼部疾病中光感受器-RPE-脉络膜神经血管单元的治疗诊断学 PI：Robert J. Zawadzki，博士概括遗传性和年龄相关性黄斑变性 (AMD) 目前是造成严重视力障碍的原因在超过 200 万美国居民中，随着人口老龄化，到 2040 年，患病率预计将翻一番。变性发生在感光细胞、视网膜色素上皮 (RPE) 细胞和脉络膜中脉管系统是眼后部紧密相互依赖的组织的复合体，该项目将继续进行。动物模型中光感受器-RPE-脉络膜复合体的形态和功能的体内研究视网膜变性，包括 RPE 线粒体功能障碍模型和 Doyne 蜂窝视网膜这些模型概括了遗传性和年龄相关性黄斑变性的主要特征： RPE 细胞变性以及 RPE 和 Bruch 之间细胞外沉积物与年龄相关的增加该项目将使用创新的细胞膜，将 RPE 与脉络膜毛细血管床分开。视网膜结构和功能的水平分辨率体内探测与附加功能测试相结合表征光感受器-RPE-脉络膜神经血管单元主要成分与年龄相关的变化 (PRC-NVU)，这些研究将在具有遗传缺陷和野生的小鼠群体中纵向进行。类型对照，喂食高脂肪饮食（以加速疾病进展）。 RPE-Bruch 膜的主要缺陷会导致光感受器的二次退化，这些研究包括光感受器结构和功能的测量（使用时间散斑平均 - 光学相干断层扫描 (TSA-OCT)、基于 OCT 的视视网膜成像 (ORG)) 和全视野视网膜电图 (ffERG))，RPE 镶嵌和布鲁赫膜的映射水驱动的 BrM 散射、RPE 细胞自发荧光和强度瞬时变化的形态使用 TSA-OCT 测量发射光谱、RPE 细胞的氧化还原状态和脉络膜毛细血管形态我们将分析这些多维纵向数据以发现潜在的结构和血管造影。 PRC-NVU 变性的功能生物标志物并评估其对疾病进展的预测价值。体内实验结束后，每只小鼠一只眼睛的脉络膜 RPE 将用配备光学相干显微镜 (OCM) 的定制高分辨率离体共焦显微镜单元和 LiveCell 阶段顶部孵化系统，可对平装的“活”RPE 细胞进行成像巩膜（以验证体内观察并评估 RPE 细胞器的荧光寿命）；每只小鼠第二只眼的视网膜将通过常规组织学和生化进行评估在光学或电子显微镜上进行测量，以验证体内形态学发现。一种通过近红外光可降解纳米颗粒将药物输送到 RPE 的新方法将用于将治疗剂局部靶向患病的 RPE 细胞，并使用空间分辨成像来确定是否治疗药物通过结合纵向、体内成像和治疗来减缓和阻止疾病进展。光学纳米疗法，这些研究将为人眼局部靶向药物输送奠定基础疾病。