Discovery of Biomarkers for Age-Related Macular Degeneration

年龄相关性黄斑变性生物标志物的发现

• 批准号: 9384653
• 负责人: DIMITRIOS MORIKIS
• 金额: $ 37.55万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9384653
• 关键词: Affinity; Age; Age related macular degeneration; Alleles; Animal Model; Antigen-Antibody Complex; Binding; Biochemical; Biological Assay; Biological Availability; Biological Markers; Bruch' s basal membrane structure; C3bi; Cell Culture Techniques; Cells; Chemicals; Clinical; Complement; Complement 3b; Complement 3d; Complement 3d Receptors; Complement Activation; Complement Factor H; Complement Membrane Attack Complex; Complex; Computing Methodologies; Data; Deposition; Detection; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Disease; Disease Progression; Docking; Drug Design; Drusen; Enzyme-Linked Immunosorbent Assay; Epithelial; Evaluation; Eye; Fluorescence; Fluorescence Spectroscopy; Genes; Goals; Half-Life; Histologic; Human; In Vitro; Inflammation; Inflammatory; Lead; Ligands; Mediating; Methods; Modeling; Monitor; Monoclonal Antibodies; Mutation; Oral; Pathogenesis; Patients; Peptides; Pharmaceutical Preparations; Play; Population; Primates; Production; Property; Protein Engineering; Proteins; Retina; Retinal; Retinal Pigments; Risk; Role; Single Nucleotide Polymorphism; Site; Specificity; Structure; Structure of retinal pigment epithelium; Surface; Testing; Thermodynamics; Tissues; Variant; Vitronectin; Work; aged; base; biomarker discovery; complement system; cost; design; efficacy testing; fluorophore; high throughput screening; human tissue; imaging properties; in vivo; link protein; macula; molecular dynamics; molecular mass; nonhuman primate; pathogen; pharmacophore; public health relevance; rapid detection; receptor; risk variant; screening; spatiotemporal; specific biomarkers; targeted treatment; therapeutic target; tissue degeneration; virtual

Project Summary/Abstract The complement system is critical for rapid detection and clearance of pathogens and immune complexes. Distinction between self and nonself is conferred by regulator proteins, which downregulate complement activation on host cells. Complement dysregulation is involved in several inflammatory conditions, including age-related macular degeneration (AMD). Deficiencies or mutations in specific complement proteins are linked to increased risk of developing AMD, and complement-targeted therapeutics have shown promise in treating AMD. In addition, it is critical to be able to monitor the onset and progression of AMD, as aberrant complement activation occurs on the retinal pigmented epithelium (RPE) in AMD patients. Complement proteins and regulators are constituents of drusen, including the terminal product C3d that remains covalently attached on surfaces on which complement has been activated. Our goal is to develop low-molecular mass chemical compounds and peptides that have strong binding affinities for C3d and fluorescence properties. These molecules will have potential as quantifiable biomarkers of complement activation in the eye and as diagnostics for early stage AMD. Our methods are computational and experimental. We will use pharmacophore-based virtual high-throughput screening, and protein-ligand docking to discover small chemical compounds with predicted binding affinities for C3d and intrinsic fluorescence. We will also use peptide design principles to design peptides with attached clinically-approved fluorophores that have predicted binding affinities for C3d. Both approaches are based on the interactions of C3d with its receptor, complement receptor 2 (CR2), and complement regulator Factor H (FH), using crystallographic structures and molecular dynamics simulations. We will test our computationally predicted C3d ligands, using an in vitro direct binding, biochemical and functional assays, and we will examine their photophysical properties using fluorescence spectroscopy. We will then evaluate the efficacies of the C3d ligands with highest potency and optimal fluorescence properties in human retinal pigmented epithelial cell-based assays. Finally, we will test the most efficacious ligands in histological sections of drusen-laden retinal tissues from non-human primate models of AMD, as well as using retinal sections from human AMD eyes. The proposed studies will generate data for a more focused study to evaluate a lead compound in animal models. !

项目总结/摘要 补体系统对于快速检测和清除病原体和免疫复合物至关重要。 自我和非自我之间的区别是由下调补体的调节蛋白赋予的 激活宿主细胞。补体失调与几种炎症性疾病有关，包括 年龄相关性黄斑变性（AMD）。特定补体蛋白的缺陷或突变与 AMD的风险增加，而补体靶向治疗已显示出治疗AMD的前景。 AMD.此外，能够监测AMD的发作和进展是至关重要的，因为异常补体 AMD患者的视网膜色素上皮（RPE）上发生活化。补体蛋白和 调节剂是玻璃疣的成分，包括保持共价连接在玻璃疣上的末端产物C3 d。 表面上的补体已被激活。我们的目标是开发低分子量化学品 对C3 d具有强结合亲和力和荧光性质的化合物和肽。这些 分子将具有作为眼睛中补体激活的可定量生物标志物的潜力， 诊断早期AMD。我们的方法是计算和实验。我们将使用 基于药效团的虚拟高通量筛选，以及蛋白质-配体对接，以发现小的 对C3 d和固有荧光具有预测结合亲和力的化合物。我们还将使用 肽设计原理，用于设计具有临床批准的荧光团的肽， 对C3 d的结合亲和力。这两种方法都是基于C3 d与其受体，补体 受体2（CR2）和补体调节因子H（FH），使用晶体结构和分子生物学方法， 动力学仿真我们将测试我们的计算预测的C3 d配体，使用体外直接结合， 生物化学和功能测定，我们将研究他们的生物物理特性，使用荧光 谱然后，我们将评估具有最高效力和最佳活性的C3 d配体的效力。 在基于人视网膜色素上皮细胞的测定中的荧光性质。最后，我们将测试最 来自非人灵长类动物模型的载有玻璃疣的视网膜组织的组织切片中的有效配体 AMD，以及使用来自人AMD眼睛的视网膜切片。拟议的研究将产生数据， 更集中的研究，以评估动物模型中的铅化合物。 !