Flavoprotein Autofluorescence Imaging in AMD

AMD 中的黄素蛋白自发荧光成像

• 批准号: 8953161
• 负责人: SCOTT W COUSINS
• 金额: $ 20.64万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2017-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8953161
• 关键词: Acute; Affect; Age related macular degeneration; Aging; Algorithms; Animals; Biological Markers; Bruch' s basal membrane structure; Cell Culture Techniques; Cells; Cessation of life; Characteristics; Chronic; Cicatrix; Clinic; Clinical; Clinical Research; Companions; Computer software; Coupled; Cultured Cells; Cytosol; Deposition; Developed Countries; Devices; Disease; Disease model; Drug Targeting; Drusen; Elderly; Electron Transport; Experimental Animal Model; Exposure to; Extracellular Matrix; Exudative age-related macular degeneration; Eye; Flavoproteins; Fluorescence Spectroscopy; Frequencies; Functional disorder; Genetic; Goals; Health; Hemorrhage; Histopathology; Human; Hydroquinones; Image; Imaging Device; Individual; Induced Mutation; Injection of therapeutic agent; Lasers; Legal Blindness; Life; Lipids; Lipofuscin; Measures; Mediating; Metabolism; Methods; Mitochondria; Mitochondrial DNA; Mus; Ophthalmoscopes; Ophthalmoscopy; Optics; Oxidants; Oxidases; Oxidative Phosphorylation; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phospholipids; Photoreceptors; Pigment Epithelium of Eye; Pilot Projects; Plasma; Population; Production; Proteomics; Reperfusion Injury; Research; Retina; Retinal; Retinal Diseases; Role; Scanning; Societies; Source; Staging; Structure of retinal pigment epithelium; Superoxides; System; Technology; Therapeutic; Time; Tissues; Toxic Environmental Substances; United States; Vascular Endothelial Growth Factors; Work; absorption; age related; base; clinical application; design; effective therapy; extracellular; geographic atrophy; human subject; hydroquinone; image processing; imaging biomarker; improved; in vivo; mitochondrial dysfunction; mouse model; novel; optical spectra; pre-clinical; programs; protein expression; prototype; public health relevance; repaired; response to injury

 DESCRIPTION (provided by applicant): In developed societies, age-related macular degeneration (AMD) is the most common cause of legal blindness in people over age 65. Emerging evidence demonstrates an important role for retinal pigment epithelium (RPE) mitochondrial dysfunction in dry AMD pathobiology. Mitochondrial dysfunction is characterized by chronic overproduction of superoxide and diminished production of ATP, and it promotes activation of injury response pathways in RPE cells, leading to sub-RPE deposit formation (precursors of lipid-rich drusen) and dysregulation of extracellular matrix turnover, the hallmark features of dry AMD. Thus, mitochondrial-targeting drugs are emerging as an attractive class of potential therapeutics for dry AMD. Technologies to identify the presence of RPE mitochondrial dysfunction in living eyes are limited. Detecting mitochondrial dysfunction in vivo would enable selection of individuals who might benefit from mitochondria-targeting drugs and would accelerate mechanistic studies of mitochondrial dysfunction in dry AMD disease models. The goal of this project is to develop RPE flavoprotein autofluorescence as an imaging biomarker for mitochondrial dysfunction. Mitochondrial dysfunction is accompanied by an increased ratio in the oxidized to reduced forms of flavoproteins (especially FAD+), which are important components of the electron transport chain for ATP production. Increase in oxidized flavoprotein produces a characteristic shift in the autofluorescence emission spectra, which is easily measured in isolated cells or tissues. However, detecting this shift noninvasively in tissues of living animals or humans, especially the eye, is much more challenging. It requires both a specialized device to image the RPE and an analytical approach to detect the specific autofluorescence signature attributable to flavoprotein. In this project, we will design and build prototype multispectral imaging device using a rapidly tunable excitation laser source, an acousto-optical tunable barrier filter, and a confocal scanning laser ophthalmoscope retinal imager, integrated with analytical software based on synchronous fluorescence spectroscopy, to detect and quantify RPE flavoprotein autofluorescence. We will use cell culture and animal model experimental systems to develop and validate this technology, and we will adapt the integrated multispectral imaging device to initiate a pilot clinical study of RPE flavoprotein autofluorescence in human subjects without retinal disease and in subjects with AMD.

 描述(申请人提供)：在发达国家，老年性黄斑变性(AMD)是65岁以上人群法定失明的最常见原因。越来越多的证据表明视网膜色素上皮(RPE)线粒体功能障碍在干性AMD的病理生物学中起着重要作用。线粒体功能障碍的特征是长期过量产生超氧化物和减少ATP的产生，并促进RPE细胞损伤反应通路的激活，导致亚RPE沉积(富含脂质的玻璃体前体)形成和细胞外基质周转失调，这是干性AMD的特征。因此，线粒体靶向药物正在成为治疗干性AMD的一类有吸引力的潜在疗法。在活体眼睛中识别RPE线粒体功能障碍的技术是有限的。在体内检测线粒体功能障碍将使选择可能受益于线粒体靶向药物的个体成为可能，并将加快干性AMD疾病模型中线粒体功能障碍的机制研究。该项目的目标是开发RPE黄素蛋白自发荧光作为线粒体功能障碍的成像生物标记物。线粒体功能障碍伴随着氧化和还原形式的黄素蛋白(特别是FAD+)比例的增加，这些黄素蛋白是ATP产生的电子传输链的重要组成部分。氧化黄素蛋白的增加会导致自发荧光发射光谱的特征位移，这在分离的细胞或组织中很容易测量到。然而，在活体动物或人类的组织中非侵入性地检测这种变化，特别是在眼睛中，要具有更大的挑战性。它既需要专门的设备来对RPE进行成像，也需要一种分析方法来检测可归因于黄素蛋白的特定自体荧光信号。在这个项目中，我们将设计和建造原型多光谱成像设备，使用快速可调的激发光源、声光可调屏障滤光片和共焦扫描激光检眼镜视网膜成像器，并集成基于同步荧光光谱的分析软件来检测和定量RPE黄素蛋白的自发荧光。我们将使用细胞培养和动物模型实验系统来开发和验证这项技术，我们将采用集成的多光谱成像设备来启动在没有视网膜疾病的人类受试者和AMD受试者中进行RPE黄素蛋白自发荧光的试点临床研究。