Auto-Antibodies as Serum Biomarkers for Age-Related Macular Degeneration

自身抗体作为年龄相关性黄斑变性的血清生物标志物

• 批准号: 7471805
• 负责人: ALESSANDRO IANNACCONE
• 金额: $ 21.89万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7471805
• 关键词: Age; Age related macular degeneration; Ancillary Study; Antibodies; Antigen Presentation; Antigen-Presenting Cells; Antigens; Award; Biological; Biological Markers; Bruch' s basal membrane structure; CD4 Positive T Lymphocytes; Choroid; Data; Dendritic Cells; Deposition; Desegregation; Development; Disease; Drusen; Elderly; Experimental Genetics; Exploratory/Developmental Grant; Eye; Frequencies; Fresh Tissue; Funding Mechanisms; Future; Gel; Health; Helper-Inducer T-Lymphocyte; Human; Immune; Immune response; Immunohistochemistry; Individual; Infiltration; Inflammation; Inflammatory; Lead; Legal Blindness; Literature; Mass Spectrum Analysis; Measurable; Mediating; Mentored Patient-Oriented Research Career Development Award; Methodological Studies; Modification; Participant; Pathogenesis; Pathway interactions; Patients; Prevention; Proteins; Recruitment Activity; Research; Research Project Grants; Resources; Retinal; Role; Sampling; Serum; Spottings; Staging; Structure of retinal pigment epithelium; Testing; Time; Tissues; base; cohort; evidence base; genetic variant; innovation; macula; prognostic; repository; research study; therapeutic target

DESCRIPTION (provided by applicant): Age-related macular degeneration (AMD) is characterized since its earliest stages by formation of deposits between the Bruch's membrane (BM) and the retinal pigment epithelium (RPE), termed drusen. In recent years, a role for inflammation in the pathogenesis of both AMD in general and drusen in particular has been established at the biological, experimental, and genetic level. Drusen have high contents of inflammatory effectors and by-products, and dendritic cells (DCs) from the choroid (Ch), which are among the resident antigen-presenting cells of the eye, are recruited to and infiltrate the BM and drusen cores. With these premises, and based on evidence from the literature indicating that AMD patients have a higher frequency of circulating antibodies directed against antigens not only of Ret, but also of BM/Ch and RPE origin than in controls, we hypothesize that auto-antibodies (Auto-Abs) are biomarkers relevant to ocular disease status and that they will be measurable in the serum of AMD patients. To test the basic tenet of this hypothesis, the Aim of this proposal is to analyze serum samples collected from participants with (n=188) and without (n=222) AMD. Of these, 108 of the AMD subjects and the 222 unaffected individuals (mean age: 79 years old) are participants in the ARMA Study, a study ancillary to Health ABC conducted by the P.I. during his K23 Award. Additional 80 subjects with advanced AMD will be recruited during this award. Serum samples collected at the time of examination will be screened for the presence of auto-Abs directed against macula-specific Ret, RPE, and BM/Ch homogenates generated from fresh tissues dissected from human donors eyes. Homogenate proteins reacting against serum auto-Abs will be immunoprecipitated and separated on 2D gels. The identity of the most common gel spots observed uniquely and/or more intensely in AMD samples will be preliminarily characterized by mass spectrometry. Based on our overall rationale, we predict that sera from AMD patients will show higher frequency of auto-Abs than control subjects not only against Ret, but also and especially against RPE and/or BM/Ch antigens. If our hypothesis will prove correct, in a subsequent R01 application we plan to test the additional hypotheses that (1) within the group of patients with AMD, anti-Ret, anti-RPE, and/or anti-BM/Ch auto-Abs will be seen more often in subjects harboring disease-predisposing genetic variants, and that (2) patients with advanced AMD will have a higher frequency of auto-Abs than patients with early-stage AMD. We further plan to (3) characterize in detail all the identified auto-antigens towards which the auto-Abs are directed by mass spectrometry and (4) localize their expression at the tissue level by conducting immunohistochemistry experiments on macular section from human donor eyes. We predict that, ultimately, this line of research will provide us with useful biomarkers to elucidate further at the tissue level the pathways involved in AMD, and to identify potential prognostic and therapeutic targets.

描述（由申请人提供）：与年龄相关的黄斑变性（AMD）的特征是由于其最早的阶段是通过Bruch膜（BM）和视网膜色素上皮（RPE）之间的沉积物形成，称为drusen。近年来，在生物学，实验和遗传水平上已经确定了炎症在AMD和DRUSEN的发病机理中的作用。 DRUSEN具有较高的炎症效应子和副产品，以及来自脉络膜（CH）的树突状细胞（DC），它们是眼睛的常驻抗原细胞之一，被募集到BM和DRUSEN核心。有了这些前提，并基于文献的证据表明，AMD患者的循环抗体的频率更高，针对RET的抗原，而且与对照组相比，与对照组相比，与对照组相比，我们的自身抗体（自动标记物（Auto-Abs））与患者相关，他们的生物标志是Ocular疾病状况和衡量AM的生物量。为了检验该假设的基本原则，该提案的目的是分析（n = 188）和没有（n = 222）AMD的参与者收集的血清样品。其中，有108位AMD受试者和222名未受影响的人（平均年龄：79岁）是ARMA研究的参与者，这是P.I.卫生ABC的一项研究。在他的K23奖中。在本奖项期间，还将招募80名具有高级AMD的主题。在检查时收集的血清样品将被筛选，以针对针对大黄斑特异性RET，RPE和BM/CH匀浆的自动AB，从人类捐助者眼中剖析的新鲜组织产生的自动AB。与血清自动abs反应的匀浆蛋白将在2D凝胶上免疫沉淀并分离。在AMD样品中观察到的最常见凝胶斑的身份将以质谱法的初步为特征。基于我们的整体基本原理，我们预测，来自AMD患者的血清不仅要比对照组，而且还针对RET，尤其是针对RPE和/或BM/CH抗原的对照受试者的频率更高。如果我们的假设证明是正确的，那么在随后的R01应用中，我们计划测试（1）在AMD，ANTI-RET，抗RPE和/或抗BM/CH自动AB的患者组中（1）在具有预期疾病的遗传变异的患者（2）患者的患者中，将更频繁地看到AMD，抗RET，抗RPE和/或抗BM/CH自动-ABS的频率更高，并且会更频繁地出现。我们进一步计划（3）详细介绍了所有已确定的自身抗原，其自动抗原由质谱指导，（4）通过在人类供体眼中的黄斑剖面上进行免疫组织化学实验，将其表达定位于组织水平。我们预测，最终，这项研究将为我们提供有用的生物标志物，以在组织水平上进一步阐明AMD中涉及的途径，并确定潜在的预后和治疗靶标。