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-螺旋神经血管单元的疗法 PI：Robert J. Zawadzki，博士概括遗传和与年龄有关的黄斑变性（AMD）目前负责严重视力障碍在超过200万的美国居民中，随着人口年龄的增长，预计到2040年的流行率将翻一番。变性发生在感光细胞，视网膜色素上皮（RPE）和脉络膜中脉管系统，后眼中紧密相互依赖的组织的复合物。这个项目将继续我们在动物模型中，光感受器-RPE-舌形成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 Honeycomb视网膜营养不良。这些模型概括了遗传和年龄相关的黄斑变性的主要标志： RPE细胞的退化以及与年龄相关的RPE和BRUCH之间的细胞外沉积膜，将RPE与脉络膜毛细血管床分开。该项目将使用创新的细胞 - 视网膜结构和功能的体内探测中的水平分辨率与其他功能测试相结合表征了年龄相关的光感受器-RPE-螺旋神经血管单元的主要成分的变化（PRC-NVU）。这些研究将在患有遗传缺陷和野生的小鼠队列中进行纵向进行类型对照，以高脂饮食喂食（加速疾病进展）。研究将检验假设 RPE-BRUCH膜中的主要缺陷导致感光器和脉络膜毛细血管脉管系统。研究包括光感受器结构和功能的度量（使用）时间斑点平均 - 光学相干断层扫描（TSA-OCT），基于OCT的Optoretinography （org））和全场电子图（FFERG），RPE Mosaic和Bruch膜的映射与水驱动的瞬态变化，BRM散射，RPE细胞自动荧光强度和发射光谱，RPE细胞的氧化还原状态以及使用TSA-OCT的绒毛膜毛细血管形态血管造影。我们将分析这些多维纵向数据，以发现潜在的结构和 PRC-NVU变性的功能生物标志物，并评估其疾病进展的预测价值。在在体内实验的终止，将用A的一只眼睛的脉络脉冲将成像定制的高分辨率外体共聚焦显微镜，配备了光学相干显微镜（OCM）单位和Livecell舞台顶孵育系统，允许对“活” RPE单元格进行成像巩膜（允许验证体内观测和RPE细胞器荧光寿命的评估）；每只小鼠的第二眼的视网膜将通过常规的组织学和生化评估在光或电子显微镜上执行的测量值，以验证体内形态学发现。通过近红外降解纳米颗粒将药物输送到RPE的新方法将用于局部靶向治疗剂，用于病态的RPE细胞，空间分辨的成像将用于确定是否是否治疗剂缓慢并停止疾病进展。通过结合纵向，体内成像和光学纳米疗法，这些研究将为人眼中的局部靶向药物提供基础疾病。