A mouse model and iPS cells to study hyperactive ABCA1 in the eye in age-related macular degeneration

小鼠模型和 iPS 细胞研究年龄相关性黄斑变性中过度活跃的 ABCA1

• 批准号: 10362536
• 负责人: Nicholas Lyssenko
• 金额: $ 19.22万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10362536
• 关键词: ATP binding cassette transporter 1; ATP-Binding Cassette Transporters; Adherent Culture; Adverse effects; Age related macular degeneration; Age-Months; Aging; Agreement; Alleles; Alzheimer' s Disease; Animal Model; Antibodies; Apolipoprotein A-I; Apolipoprotein E; Apolipoproteins; Apolipoproteins B; Appearance; Area; Atherosclerosis; Blood Circulation; Blood capillaries; Bruch' s basal membrane structure; Cell Culture Techniques; Cell Line; Cell membrane; Cells; Cholesterol; Choroid; Complex; Data; Deposition; Development; Diagnostic; Disease; Drusen; Environment; Exhibits; Extracellular Matrix; Exudative age-related macular degeneration; Eye; Fundus photography; Genetic study; Goals; High Density Lipoprotein Cholesterol; High Density Lipoproteins; Human; Human Genetics; Hyperactivity; Individual; Investigation; LXRalpha protein; Lesion; Lipids; Lipoproteins; Mediating; Mifepristone; Minor; Morphology; Mus; Mutation; Oils; Pathology; Phospholipids; Plasma; Production; Protein Isoforms; Proteins; Resources; Retina; Risk; Risk Factors; Role; Side; Source; Stains; Structure; Structure of retinal pigment epithelium; System; Testing; Therapeutic; Time; Vesicle; Virulence Factors; Water; basolateral membrane; beta catenin; experimental study; genome wide association study; geographic atrophy; human fetal retinal pigment epithelial cell; in vivo; induced pluripotent stem cell; macula; member; monolayer; mouse model; neovascularization; overexpression; particle; promoter; repository; water flow

Abstract Age-related macular degeneration (AMD) is a disease of aging that presently has very limited therapeutic resources. Soft drusen are deposits of protein and lipid between the retinal pigment epithelium (RPE) and choroidal capillaries in the Bruch's membrane. These deposits are visible in fundus photography as white to yellow patches in the macula. Their presence is a diagnostic feature of early asymptomatic AMD and a risk factor for developing late, symptomatic AMD. It was suggested over 25 years ago that drusen interfere with the flow of water and metabolites between the choroid capillaries and RPE cells and cause RPE cells either to die as in geographic atrophy and to initiate neovascularization as in wet AMD. However, the source of druse lipid has remained unknown. At various times, it was proposed that RPE plasma membrane vesicles or apolipoprotein B-lipoprotein secreted by the RPE was the source of these lesions. Recent genome-wide association studies pointed to an unexpected culprit: high-density lipoprotein (HDL). It had been known that apolipoprotein E (apo E) isoform ε4 is protective for AMD. This is the same isoform that does not mediate cell cholesterol efflux well and is a risk factor in atherosclerosis and Alzheimer's disease. But in AMD it is protective. The genome-wide association studies additionally implicated ATP-binding cassette transporter subfamily A member 1 (ABCA1) in AMD. ABCA1 mediates synthesis of HDL from lipid and apoE. ABCA1 alleles that increase plasma HDL cholesterol levels are a risk factor for AMD. Thus, increased production of HDL is an AMD rick factor. We outline reasons from druse composition and our preliminary data for the RPE and not plasma origins of HDL that is deposited in drusen. We then propose to test the hypothesis that hyperactive synthesis of HDL by the RPE contributes to druse formation. The hypothesis is tested in a mouse model of ABCA1 overexpression in the RPE in Aim 1. The hypothesis is also tested in polarized RPE monolayer cultures derived from human induced pluripotent stem cells either expressing or missing ABCA1 in Aim 2. Results of this exploratory project will provide evidence for a larger project into the role of HDL in AMD pathology.

抽象的 与年龄相关的黄斑变性（AMD）是一种衰老的疾病，目前的治疗非常有限 资源。软drusen是视网膜色素上皮（RPE）之间的蛋白质和脂质的沉积物 Bruch膜中的脉络膜毛细血管。这些沉积物在底眼摄影中可见 黄斑中的黄色斑块。它们的存在是早期无症状AMD的诊断特征和风险 迟到，有症状的AMD的因素。 25年前，有人建议杜鲁森干扰 脉络膜毛细血管和RPE细胞之间的水和代谢产物的流动，并导致RPE细胞死亡 就像在地理萎缩中一样，启动新血管形成，如湿AMD中一样。但是，Druse脂质的来源 仍然未知。在不同时间，有人提出RPE质膜蔬菜或 RPE分泌的载脂蛋白B-脂蛋白是这些病变的来源。最近全基因组 协会研究指出了意外的罪魁祸首：高密度脂蛋白（HDL）。众所周知 载脂蛋白E（APO E）同工型ε4受到AMD的保护。这是不介导细胞的同工型 胆固醇外排很好，是动脉粥样硬化和阿尔茨海默氏病的危险因素。但是在AMD中是 保护的。全基因组关联研究还实施了ATP结合盒转运蛋白 AMD中的亚家族成员1（ABCA1）。 ABCA1介导了脂质和APOE的HDL合成。 ABCA1 增加血浆HDL胆固醇水平的等位基因是AMD的危险因素。这增加了生产 HDL是AMD Rick因子。我们概述了Druse组成的原因以及RPE的初步数据 而不是沉积在drusen中的HDL的血浆起源。然后，我们建议检验以下假设 RPE对HDL的多动性合成有助于Druse的形成。该假设在小鼠中进行了检验 AIM 1中RPE中ABCA1过表达的模型。在极化RPE中还测试了该假设 源自人类诱导的多能干细胞的单层培养物要么表达或缺失ABCA1 AIM 2。此探索项目的结果将为HDL在AMD中的作用提供更大的项目的证据 病理。