Soft drusen in rhesus macaques as a nonhuman primate model of early age-related macular degeneration

恒河猴的软玻璃疣作为早期年龄相关性黄斑变性的非人类灵长类动物模型

• 批准号: 10547804
• 负责人: Glenn Yiu
• 金额: $ 68.48万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10547804
• 关键词: Age; Age related macular degeneration; Animal Model; Animals; Apolipoprotein E; Apolipoproteins; Apolipoproteins B; Arterial Fatty Streak; Binding; Bioinformatics; Blindness; Body Weight; Body Weights and Measures; Body fat; California; Cardiovascular Diseases; Cells; Cholesterol; Cholesterol Esters; Choroid; Complement; Complement Membrane Attack Complex; Contralateral; Deposition; Development; Diet; Dietary Fats; Dose; Drusen; Elderly; Epidemiology; Euthanasia; Evolution; Exposure to; Eye; Fasting; Fatty acid glycerol esters; Fundus; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Predisposition to Disease; Glial Fibrillary Acidic Protein; Goals; Growth; Health; Heterogeneity; High Density Lipoproteins; High Fat Diet; Histologic; Human; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Image; Imaging technology; Immune; Immunohistochemistry; Inflammatory Response; Intervention; Laboratory Animals; Lipids; Lipofuscin; Lipoproteins; Low-Density Lipoproteins; Macaca; Macaca mulatta; Macrophage Activation; Mammals; Measures; Metabolic; Modeling; Molecular; Multimodal Imaging; National Institute on Aging; Oils; Optical Coherence Tomography; Oral; Pathogenesis; Pathway interactions; Patients; Phenotype; Pilot Projects; Placebos; Plasma; Prevalence; Primates; Research; Resolution; Retina; Risk Factors; Role; Serum; Simvastatin; Smoking; Structure of retinal pigment epithelium; Surveys; Susceptibility Gene; Testing; Tissues; Transferase; Triglycerides; aged; atorvastatin; cohort; cost; dietary; genetic variant; genome sequencing; hepatic lipase; imaging capabilities; in vivo; in vivo imaging; insight; lipid metabolism; lipoprotein lipase; mRNA Expression; macroglia; macula; multimodality; new therapeutic target; next generation sequencing; nonhuman primate; novel therapeutics; ophthalmic examination; oxidation; programs; recruit; serial imaging; sex; single-cell RNA sequencing; sugar; therapy development; translational study; whole genome

PROJECT SUMMARY Age-related macular degeneration (AMD) is the leading cause of vision loss in the elderly, but current treatments target advanced stages when interventions may be limited by irreversible cell loss. Soft drusen are the initial and hallmark feature of AMD, but their pathogenesis is unclear and the development of treatments is limited by the lack of good animal models. Unlike most laboratory animals, nonhuman primates (NHPs) possess a true macula and spontaneously develop soft drusen resembling early human AMD. Prior studies of NHP drusen have been observational, cross-sectional, and qualitative, owing to the high costs of maintaining aged animals, limited longitudinal imaging capabilities, and lack of Next-Generation Sequencing (NGS) and bioinformatics support. We recently surveyed a unique colony of geriatric rhesus macaques at the California National Primate Research Center (CNPRC), and found a 30.6% prevalence of drusen. Using high-resolution multimodal in vivo imaging, we precisely measured drusen progression over 2 years, with histological and ultrastructural correlates demonstrating lipid accumulation resembling human soft drusen. Similar to cardiovascular diseases, AMD share epidemiologic risk factors such as high fat diets, susceptibility genes in lipid metabolism, and deposits similar to atherosclerotic plaques with lipoprotein accumulation, lipid oxidation, and complement-based inflammatory responses. Recent pilot studies suggest that high-dose oral atorvastatin can cause drusen regression in some AMD patients, but human studies are limited by the heterogeneity of AMD phenotypes and variability in subject diets and serum lipid concentrations. In this study, we plan to establish a new, unique cohort of aged rhesus macaques with soft drusen, and perform 1) detailed ophthalmic characterization using spectral-domain optical coherence tomography (SD-OCT) to precisely measure retinal layers and track drusen volume, and fundus autofluorescence (FAF) to measure RPE lipofuscin, 2) metabolic profiling to measure fasting plasma metabolites, lipoproteins, and apolipoproteins, and 3) whole-genome sequencing to identify genetic variants in animals with drusen that may be shared with human AMD. Next, we will establish the impact of a “Western-style” high-fat/sugar (HFS) diet and high-dose atorvastatin on drusen progression and RPE health over 2 years, and determine possible correlations with dietary fats and serum lipids. Finally, we will employ NGS-based single-cell RNA-sequencing (scRNA-Seq) to determine the gene expression profile in macular RPE cells with and without soft drusen, exposure to HFS diet, or treatment with high-dose statin, followed by immunohistochemistry of contralateral eyes to evaluate and validate identified gene sets or pathways, as well as complement and other immune pathways. Together, these studies will not only establish a highly-valuable and unique cohort of aged rhesus macaques with soft drusen suitable for translational studies, but also explore the role of dietary lipids, oral statins, RPE gene expression, and immune pathways in this NHP model of early AMD.

项目摘要 与年龄相关的黄斑变性（AMD）是古老的视力丧失的主要原因，但是当前治疗 当干预措施受到不可逆细胞损失的限制时，目标高级阶段。软drusen是最初的 AMD的标志性特征，但它们的发病机理尚不清楚，治疗的发展受到 缺乏好的动物模型。与大多数实验室动物不同，非人类灵长类动物（NHP）具有真正的黄斑 赞助商自发地发展出类似于早期人类AMD的软drusen。 NHP drusen的先前研究已经 观察性，横截面和定性，由于维持老年动物的高成本， 纵向成像功能，缺乏下一代测序（NGS）和生物信息学支持。 我们最近调查了加利福尼亚国家灵长类动物研究的独特老年恒河猴的独特殖民地 中心（CNPRC），发现drusen患病率为30.6％。使用高分辨率的多模式在体内成像， 我们在2年内仔细测量了drusen的进展，组织学和超微结构相关 表现出类似于人类软drusen的脂质积累。类似于心血管疾病，AMD分享 流行病学危险因素，例如高脂肪饮食，脂质代谢中的易感基因以及类似的沉积物 具有脂蛋白积累，脂质氧化和补体炎症性的动脉粥样硬化斑块 回答。最近的试点研究表明，高剂量的口服阿塔伐他汀会在某些人中引起drusen回归 AMD患者，但人类研究受到AMD表型的异质性和受试者的可变性的限制 饮食和血清脂质浓度。 在这项研究中，我们计划建立一个新的，独特的老年恒河猕猴，并进行柔软的杜鲁森，并执行 1）使用光谱域光学相干断层扫描（SD-OCT）详细的眼科表征 精确测量的残留层和跟踪drusen的体积和眼底自动荧光（FAF），以测量RPE 脂肪霉素，2）代谢分析，以测量禁食血浆代谢物，脂蛋白和载脂蛋白，以及 3）全基因组测序以鉴定具有drusen动物的遗传变异，可以与人共享 AMD。接下来，我们将建立“西式”高脂/糖（HFS）饮食和大剂量atorvastatin的影响 关于drusen的进展和RPE健康，并确定与饮食脂肪和 血清脂质。最后，我们将采用基于NGS的单细胞RNA-Sequent（SCRNA-Seq）来确定 有或没有软drusen的黄斑RPE细胞中的基因表达谱，暴露于HFS饮食或治疗 用高剂量他汀类药物，然后进行对侧眼的免疫组织化学，以评估和验证已确定的 基因组或途径，以及补体和其他免疫途径。在一起，这些研究不会 仅建立一个高度可观且独特的老年恒河猕猴，适合软drusen 翻译研究，但还探讨了饮食脂质，口服他汀类药物，RPE基因表达和免疫的作用 在这种NHP模型的早期AMD模型中。