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.

长期目标是了解眼睛中补体信号的基本基础，以及如何 在这个过程中的失调导致病理学，最终有助于治疗性AP的发展， 对毁灭性致盲疾病的研究BrM是一种五层细胞外基质隔室，含有 中间弹性层（含有弹性蛋白，EL）。这种EL的厚度和完整性更薄且含量更少 在黄斑中比在周边中更严重;在早期AMD和活动性脉络膜中的差异更严重。 新生血管形成（CNV）。AMD患者具有升高的血清EL-肽和抗EL抗体水平;以及 EL-肽可以增加脉络膜内皮细胞迁移，总体上表明EL异常 稳态在AMD中起作用。我们已经描述了烟雾诱导的眼部病理学的小鼠模型 （SIOP）。这些小鼠表现出抗EL抗体（Abs，IgG 2a）的血清水平增加， 并且显示出弹性层的完整性的完全丧失。相反，用氧化EL免疫的小鼠 当与免疫的那些相比时，（新表位）小鼠在BrM中发展更严重的视力丧失和病理学 对照EL（自身蛋白）。我们的总体假设是，年龄和/或压力依赖 弹性蛋白酶活性增加，导致弹性蛋白降解和（抗）α-弹性蛋白Ab的协同产生 是AMD的早期事件。我们进一步假设，α-弹性蛋白抗体与FcγRs的结合触发抗体- 依赖性细胞介导的细胞毒性，加剧AMD病理，这是一种由复合物放大的机制， 补体依赖性细胞毒性。此外，我们认为α-1抗胰蛋白酶（A1 AT），一种内源性弹性蛋白酶， 抑制剂可用于保持BrM完整性，作为潜在的早期AMD治疗。三个具体目标是 旨在确定小鼠模型中BrM病理学中抗弹性蛋白抗体的参与，和 第二，测试A1-AT减少小鼠和人的病理生物学的预测，使用A1-AT治疗， 在小鼠模型中进行研究，并分析MarketScan数据库中的A1-AT使用和AMD发作。 这一想法的新奇之处在于弹性蛋白酶活性导致结构病理变化（弹性蛋白损失， 炎症），并且将测试现有弹性蛋白酶抑制剂的预炎症能力是高度创新的。 服务于BrM完整性，作为一种治疗模式，在过程的早期减缓疾病的进展。