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: 7895584
• 负责人: ALESSANDRO IANNACCONE
• 金额: $ 18.5万
• 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 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7895584
• 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）：年龄相关性黄斑变性（AMD）从其最早阶段开始就以在布鲁赫膜（BM）和视网膜色素上皮（RPE）之间形成沉积物（称为玻璃膜疣）为特征。近年来，炎症在AMD和玻璃疣发病机制中的作用已在生物学、实验和遗传水平上得到证实。玻璃疣具有高含量的炎症效应物和副产物，来自脉络膜 (Ch) 的树突状细胞 (DC)（属于眼睛常驻抗原呈递细胞）被募集并渗透到 BM 和玻璃疣核心。基于这些前提，并基于文献证据表明，与对照组相比，AMD 患者不仅针对 Ret 抗原，而且还针对 BM/Ch 和 RPE 来源的抗原，具有更高频率的循环抗体，我们假设自身抗体（ Auto-Abs）是与眼部疾病状态相关的生物标志物，可以在 AMD 患者的血清中进行测量。为了检验这一假设的基本原理，本提案的目的是分析从患有 (n=188) 和不患有 (n=222) AMD 的参与者那里收集的血清样本。其中，108 名 AMD 受试者和 222 名未受影响的个体（平均年龄：79 岁）参加了 ARMA 研究，该研究是由 P.I. 进行的 Health ABC 的辅助研究。在获得 K23 奖期间。在此奖励期间还将招募另外 80 名患有高级 AMD 的受试者。检查时收集的血清样本将被筛选是否存在针对黄斑特异性 Ret、RPE 和 BM/Ch 匀浆的自身抗体，这些匀浆是从人类捐赠者眼睛解剖的新鲜组织中产生的。与血清自身抗体反应的匀浆蛋白将被免疫沉淀并在 2D 凝胶上分离。在 AMD 样品中独特和/或更强烈地观察到的最常见凝胶点的身份将通过质谱法进行初步表征。根据我们的总体原理，我们预测 AMD 患者的血清将比对照受试者显示更高频率的自身抗体，不仅针对 Ret，而且尤其针对 RPE 和/或 BM/Ch 抗原。如果我们的假设被证明是正确的，在随后的 R01 应用中，我们计划测试其他假设：(1) 在 AMD、抗 Ret、抗 RPE 和/或抗 BM/Ch 自身抗体患者组中在携带疾病易感基因变异的受试者中更常见，并且 (2) 晚期 AMD 患者比早期 AMD 患者具有更高的自身抗体频率。我们进一步计划（3）详细描述所有已识别的自身抗原，通过质谱法将自身抗体针对这些抗原，并（4）通过对人类供体眼睛的黄斑切片进行免疫组织化学实验，将其表达定位在组织水平。我们预测，最终，这一系列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 所涉及的途径，并确定潜在的预后和治疗靶点。

项目成果

Retinal pigment epithelium and microglia express the CD5 antigen-like protein, a novel autoantigen in age-related macular degeneration.

视网膜色素上皮和小胶质细胞表达 CD5 抗原样蛋白，这是年龄相关性黄斑变性中的一种新型自身抗原。

• 发表时间: 2017-02
• 影响因子: 3.4
• 作者: Iannaccone, Alessandro;Hollingsworth, T J;Koirala, Diwa;New, David D;Lenchik, Nataliya I;Beranova;Gerling, Ivan C;Radic, Marko Z;Giorgianni, Francesco
• 通讯作者: Giorgianni, Francesco