Elastase and Elastin Peptide Activity in Age-Related Macular Degeneration

年龄相关性黄斑变性中的弹性蛋白酶和弹性蛋白肽活性

• 批准号: 10077557
• 负责人: Baerbel Rohrer
• 金额: $ 32.43万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10077557
• 关键词: Acute; Age; Age of Onset; Age related macular degeneration; Air; Animal Disease Models; Animal Model; Antibodies; Antibody Formation; Autoimmune Responses; B-Lymphocytes; Basement membrane; Binding; Blindness; Bruch' s basal membrane structure; Cell model; Cell surface; Cell-Mediated Cytolysis; Choroidal Neovascularization; Complement; Complement 1q; Complement Activation; Complement-Dependent Cytotoxicity; Data; Databases; Disease; Disease Progression; Elastases; Elastin; Endothelial Cells; Enzymes; Epithelial; Event; Exhibits; Extracellular Matrix; Eye; Goals; Health; Homeostasis; Immunize; Immunoglobulin G; Inflammation; Injury; Lasers; Lectin; Lesion; Ligands; Lung; Modeling; Modification; Molecular Abnormality; Mus; Ocular Pathology; Oxidative Stress; Oxides; Pathologic; Pathology; Pathway interactions; Patients; Peptides; Play; Process; Production; Proteins; Pulmonary Emphysema; Rag1 Mouse; Reporting; Role; Serum; Signal Transduction; Smoke; Stress; Structure; Surface; T-Lymphocyte; Testing; Therapeutic; Therapeutic Effect; Thick; Transgenic Mice; Trypsin; Vascular Endothelial Growth Factors; Vision; alpha 1-Antitrypsin; antibody-dependent cell cytotoxicity; base; cell motility; complement system; design; elastase inhibitor; experimental study; exposure to cigarette smoke; gene complementation; human subject; immunogenic; innovation; intraperitoneal; lung injury; macula; man; mouse model; neoantigens; preservation; receptor; response; therapeutic development

The long-term goal is to understand the fundamental basis of complement signaling in the eye, and how misregulation in this process leads to pathology, to ultimately aid in the development of therapeutic ap- proaches for devastating blinding diseases. BrM, a pentalaminar extracellular matrix compartment, contains a middle elastic layer (contains elastin, EL). Thickness and integrity of this EL is thinner and less abundant in the macula than in the periphery; a discrepancy that is more severe in early AMD and active choroidal neovascularization (CNV). AMD patients have elevated serum EL-peptide and anti-EL antibody levels; and EL-peptides can increase choroidal endothelial cell migration, overall suggesting that abnormalities in EL homeostasis play a role in AMD. We have characterized a mouse model of smoke-induced ocular pathology (SIOP) in C57BL/6J mice. These mice exhibit increased serum levels of anti-EL antibodies (Abs, IgG2a), and show a complete loss of integrity of the elastic layer. Conversely, mice immunized with oxidized EL (neoepitope) develop more severe vision loss and pathology in BrM when compared to those immunized with control EL (self-protein). We are guided by our overall hypothesis that age- and/or stress-dependent increase in elastase activity leading to elastin degradation and coordinated production of (anti) α-elastin Abs are early events in AMD. We further hypothesize that α-elastin Abs binding to FcγRs trigger antibody- dependent cell-mediated cytotoxicity, exacerbating AMD pathology, a mechanism that is amplified by com- plement-dependent cytotoxicity. Also, we suggest that alpha-1 antitrypsin (A1AT), an endogenous elastase inhibitor, can be utilized to retain BrM integrity as a potential early AMD therapy. Three specific aims are designed to determine the involvement of antibodies against elastin in BrM pathology in mouse models, and second to test the prediction that A1-AT reduces pathobiology in mouse and man, using A1-AT for thera- peutic purposes in mouse models and analyzing the MarketScan Database for A1-AT use and AMD onset. The novelty of the idea is that elastase activity results in structural pathological changes (elastin loss, in- flammation), and it is highly innovative that an existing elastase inhibitor will be tested for its ability to pre- serve BrM integrity, as a treatment paradigm to slow down the progression of disease early in the process.

长期目标是了解眼中完成信号的基本基础，以及如何 在这一过程中的正调导致病理学，最终有助于发展治疗的发展 为毁灭性的盲目疾病提供侵害。 BRM，一种细胞外基质室，包含 中间弹性层（包含弹性蛋白，EL）。该EL的厚度和完整性更薄，较少 在黄斑中比在外围;在AMD早期和主动脉络膜中更严重的差异 新血管形成（CNV）。 AMD患者的血清EL肽和抗EL抗体水平升高。和 EL肽可以增加脉络膜内皮细胞迁移，总体表明EL异常 体内平衡在AMD中发挥作用。我们表征了烟雾诱导的眼病理的小鼠模型 （SIOP）在C57BL/6J小鼠中。这些小鼠暴露了抗EL抗体的血清水平升高（ABS，IgG2a）， 并显示弹性层完整性的完全丧失。相反，用氧化EL免疫的小鼠 （NeoEpitope）与免疫的人相比，BRM的视力丧失和病理更为严重 与控制EL（自蛋白）。我们的总体假设指导我们年龄和/或压力依赖性 弹性蛋白酶活性的增加导致弹性蛋白降解和（抗）α-肾上腺素ABS的协调产生 是AMD的早期事件。我们进一步假设α-肾上腺素与FcγR的结合触发抗体 - 依赖性细胞介导的细胞毒性，加剧AMD病理学，这种机制被调解而扩大 植物依赖性细胞毒性。另外，我们建议α-1抗胰蛋白酶（A1AT），一种内源弹性酶 抑制剂可以用来保留BRM完整性作为潜在的早期AMD治疗。三个具体目标是 旨在确定针对弹性蛋白在BRM病理学中的抗体参与小鼠模型，以及 其次测试A1-AT降低小鼠和人的病理生物学的预测，使用A1-AT进行thera- 在鼠标模型中的实现目的，并分析MarketScan数据库的使用和AMD发作。 这个想法的新颖性是弹性酶活性会导致结构性病理变化（弹性蛋白丧失，in- 燃烧），高度创新的是，现有的弹性酶抑制剂将获得预先测试的能力 将BRM的完整性作为治疗范式，以减慢此过程的早期疾病进展。