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：

• 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
• 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' 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

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：光感受器-RPE-脉络膜神经血管单位在眼科疾病中的治疗诊断学 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）细胞和脉络膜细胞中。 血管系统是后眼中紧密相互依赖的组织的复合体。该项目将继续我们在 视网膜色素上皮-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 Honeycomb视网膜 营养不良。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细胞的退化和RPE和Bruch’s之间细胞外沉积物的年龄相关性增加 膜，其将RPE与脉络膜毛细血管床分开。该项目将使用创新的蜂窝- 视网膜结构和功能的体内探测与附加功能测试相结合的水平分辨率， 表征光感受器-RPE-脉络膜神经血管单位主要成分的年龄相关变化 （PRC-NVU）。这些研究将在具有遗传缺陷和野生型的小鼠队列中纵向进行。 型对照，喂食高脂肪饮食（以加速疾病进展）。这些研究将检验这一假设 RPE-Bruch膜中的主要缺陷导致光感受器的二次退化， 绒膜毛细血管系统。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的视网膜光学成像 （ORG））和全视野视网膜电图（ffERG）），RPE镶嵌和Bruch膜的映射 在BrM散射、RPE细胞自发荧光强度和 TSA-OCT检测视网膜色素上皮细胞的发射光谱、氧化还原状态和脉络膜毛细血管形态 动脉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变性的功能性生物标志物，并评估其对疾病进展的预测价值。在 在体内实验结束时，来自每只小鼠的一只眼睛的脉络膜-RPE将用 定制的高分辨率离体共聚焦显微镜，配备光学相干显微镜（OCM） 单元和LiveCell载物台顶部孵育系统，其允许用该装置平装的“活”RPE细胞成像。 巩膜（允许验证体内观察和评估RPE细胞器的荧光寿命）; 将通过常规的组织学和生物化学方法评价每只小鼠第二只眼睛的视网膜 在光学或电子显微镜上进行的测量，以允许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递送到RPE的新方法将被用于 局部靶向治疗剂到患病的RPE细胞，空间分辨成像将用于确定是否 所述治疗剂减缓并阻止疾病进展。通过将纵向、体内成像和 光学纳米治疗，这些研究将奠定了基础，局部靶向药物输送在人类眼睛 疾病