Diagnostic classifiers for sarcoidosis using a novel T7 phage display technology

使用新型 T7 噬菌体展示技术的结节病诊断分类器

• 批准号: 9805512
• 负责人: Lobelia Samavati
• 金额: $ 11.55万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9805512
• 关键词: Affect; African American; Age; Antibodies; Antigens; Autoantibodies; Bacterial Antigens; Bacteriophage T7; Bacteriophages; Biological Markers; Biopsy; Blood Tests; Bronchoalveolar Lavage; Cells; Clinical; Clinical Data; Collection; Complex; Cutaneous; Data; Development; Diagnosis; Diagnostic; Differential Diagnosis; Disease; Embryo; Enrollment; Epitopes; Etiology; Eye; Female; Fibroblasts; Future; Granuloma; Granulomatous; Human; Hypergammaglobulinemia; Immune System Diseases; Immunity; Immunoassay; Immunoglobulin G; Immunotherapy; Inflammatory; Leukocytes; Libraries; Lung; Mediastinal lymph node group; Messenger RNA; Methods; Modeling; Neuraxis; Organ; Pathogenesis; Patients; Performance; Phage Display; Plasma; Play; Population; Population Control; Predictive Value; Pulmonary Sarcoidosis; Role; Sampling; Sarcoidosis; Sensitivity and Specificity; Serological; Skin; Technology; Testing; Tissues; Training; United States National Institutes of Health; Validation; Viral Antigens; aged; anergy; base; biobank; biomarker panel; cDNA Library; case control; cohort; diagnostic biomarker; diagnostic panel; disease diagnosis; immune function; insight; instrument; monocyte; novel; novel therapeutics; patient response; predict clinical outcome; treatment response

Abstract Sarcoidosis is an inflammatory disease of unknown etiology that occurs worldwide and is characterized by granuloma formation in different organs. No specific test has been developed to diagnose this disease. Confirmation of non-caseating granuloma in tissue biopsy of involved organs in the absence of other causes is the current state of the art for diagnosing sarcoidosis. We propose to test the hypothesis that overall immunity plays a prominent role in the pathogenesis of sarcoidosis, since abnormalities of the immune function and the presence of various antibodies/autoantibodies occurs in this disorder. Using a high throughput method, we have developed a complex epitope library derived from materials of sarcoidosis patients. The epitopes are derived from a T7 phage cDNA library of potential sarcoidosis antigens using mRNA isolated from bronchoalveolar (BAL) cells and white blood cells (WBCs) of patients with sarcoidosis. This cDNA library containing large numbers of epitopes has been immunoscreened with sera from patients with sarcoidosis containing high titer IgG antibodies and the cloned phages have been used to construct an antigen microarray to detect antibodies against sarcoid antigen(s) in the sera of test subjects. We have identified a panel of biomarkers/classifiers with high sensitivity and specificity that can discriminate between sera of patients with sarcoidosis and healthy controls. In our study we used 80-90% of African American female population of sarcoidosis patients and these patients were not strictly age-matched with healthy controls. To test this hypothesis, we propose to use banked plasma from diversified population of sarcoidosis patients and aged-matched healthy controls, enrolled in the NIH-sponsored A Case Controlled Etiology of Sarcoidosis Study (ACCESS). We would like to use these plasma samples and the clinical data from ACCESS biorepository to first test and validate the bioreactivity of plasma obtained independently from cases and controls (ACCESS) to obtain a panel of diagnostic biomarkers/classifiers, which can discriminate between sarcoidosis and healthy controls. Second, we will determine whether the discovered biomarkers/classifiers can predict the clinical outcome and Scadding stages of sarcoidosis. In future, this approach could be used to identify a panel of biomarkers useful for diagnosis of various organ involvements in sarcoidosis and differential diagnosis of various granulomatous diseases or response to treatment in sarcoidosis subjects.

摘要 结节病是一种病因不明的炎性疾病，在世界范围内发生， 以在不同器官中形成肉芽肿为特征。尚未制定具体测试 来诊断这种疾病受累组织活检证实非干酪化性肉芽肿 在没有其他原因的情况下检查器官是诊断结节病的现有技术。 我们建议检验这一假设，即整体免疫力在免疫系统中起着重要作用。 结节病的发病机制，因为免疫功能异常和存在 各种抗体/自身抗体出现在这种疾病中。使用高通量方法，我们 已经开发了一个来源于结节病患者材料的复合表位文库。的 表位来自潜在结节病抗原的T7噬菌体cDNA文库， 从支气管肺泡（BAL）细胞和白色血细胞（WBC）分离的mRNA， 结节病这个含有大量抗原表位的cDNA文库已经过免疫筛选 用含有高滴度IgG抗体的结节病患者血清和克隆的 已将其用于构建检测类肉瘤抗体的抗原微阵列 测试对象血清中的抗原。我们已经确定了一组生物标志物/分类器， 可区分结节病患者血清的高灵敏度和特异性 健康的对照。在我们的研究中，我们使用了80-90%的非洲裔美国女性人口， 结节病患者，这些患者与健康对照组的年龄不严格匹配。到 为了验证这一假设，我们建议使用来自不同人群的库存血浆， 结节病患者和年龄匹配的健康对照，入组NIH赞助的A病例 结节病病因对照研究（ACCESS）。我们想用这些血浆 样品和来自ACCESS生物储存库的临床数据， 从病例和对照（ACCESS）中独立获得的血浆的生物活性，以获得 一组诊断生物标志物/分类器，可以区分结节病和 健康对照其次，我们将确定所发现的生物标志物/分类器是否可以 预测结节病的临床结局和Scadding分期。在未来，这种方法可以 用于识别一组可用于诊断各种器官受累的生物标志物 结节病和各种肉芽肿性疾病的鉴别诊断或对 治疗结节病受试者。