High-throughput immunoproteomics for cancer biomarker discovery

项目摘要/摘要 亚利桑那州立大学生物标志物鉴定中心的目标是通过以下方式改善卵巢癌和肺癌的筛查 与生物相关的循环免疫生物标记物的发展。我们中心的科学方法是 基于几个基本原则。首先，这改变了癌症蛋白的表达、结构和后- 翻译修饰诱导宿主自身抗体产生循环生物标记物。第二，这种变化 在微生物中表达抗原(如呼吸道病原体)也可诱导免疫，常见于良性疾病 而不是恶性疾病。第三，蛋白质的修饰，以及对这些的免疫反应 新的抗原性结构，人与人之间是不同的，血清生物标记物可以补充 循环蛋白生物标志物。我们将采用系统免疫学的方法来发现三种类型的 抗体、抗微生物抗体、自身抗体和抗异常糖蛋白抗体。我们的建议 以我们在癌症生物标记物发现和免疫蛋白质组学技术方面的丰富经验为基础 发展。我们之前关于自身抗体生物标记物的结果已经在盲法第二阶段得到证实。 多中心验证研究，并导致了CLIA认证的商业血液测试。我们的结果表明， 要有足够的预测价值，需要多组自身抗体。有了先前的EDRN支持，我们 开发了一套创新的免疫蛋白质组学技术，即高密度核酸 可编程蛋白质阵列(HD-NAPPA)、反向捕获蛋白质阵列(CCPA)和在溶液中多路传输 蛋白质阵列(MISPA)，连同我们最大的全长人类和微生物基因收集 DNASU质粒库，使我们能够研究针对完整的人类蛋白质组、微生物蛋白质组的抗体 和人类O-糖蛋白在癌症中的抗体生物标记物签名。我们的中尺度诊断 (MSD)团队在全球部署了3,000多台仪器，以及700多种可商业化的生物标志物分析 工具包。我们在血清学分析开发方面的专业知识被Warp Fast选中使用V-PLEX® 作为其所有资助阶段免疫原性评估的标准结合分析的基础的血清学小组 新冠疫苗临床试验。我们将使用我们的MSD多阵列平台来迁移顶级血清学和 蛋白质标记物在我们的目标临床应用中的效用。我们将与肺和肺方面的专家合作 波士顿大学范德比尔特大学医学中心的卵巢癌筛查 中心和德国癌症研究中心，谁也将提供高质量的 用于开发循环生物标志物的样本，以加强卵巢癌筛查或区分良恶性 恶性肺结节。秉承探测器设计原则，我们将通过以下方式执行第一阶段发现 用癌症患者和对照血清筛选蛋白质阵列以检测癌症或对照特异性抗体。侯选人 肺癌和卵巢癌的生物标记物都将经历第二阶段的验证。

Project Summary/Abstract The goal of the ASU Biomarker Characterization Center is to improve ovarian and lung cancer screening through the development of biologically-relevant circulating immune biomarkers. The scientific approach of our Center is based on several fundamental principles. First, that altered cancer protein expression, structure, and post- translational modifications induce host autoantibodies to create circulating biomarkers. Second, that alterations in microbial antigen expression (such as respiratory pathogens) also induce immunity, often detected in benign rather than malignant disease. Third, that the protein modifications, as well as the immune response to these neoantigenic structures, are heterogeneous between people, and that serologic biomarkers may complement circulating protein biomarkers. We will take a systems immunology approach to discover three types of antibodies, anti-microbial antibodies, autoantibodies and anti-aberrant glycoprotein antibodies. Our proposal builds on our extensive experiences with cancer biomarker discovery and immunoproteomics technology development. Our previous results on autoantibody biomarkers have been confirmed in blinded phase 2 multicenter validation studies and led to a CLIA-certified commercial blood test. Our results have shown that multiplexed panels of autoantibodies are required for adequate predictive value. With prior EDRN support, we have developed a set of innovative immunoproteomics technologies, namely high-density nucleic acid programmable protein array (HD-NAPPA), contra-capture protein array (CCPA) and multiplexed in solution protein array (MISPA), that, together with the largest full-length human and microbial gene collection at our DNASU plasmid repository, enable us to study antibodies against the full human proteome, microbial proteomes and the human O-glycoproteome for antibody biomarker signatures in cancer. Our Meso Scale Diagnostics (MSD) team has fielded over 3,000 instruments worldwide, and over 700 commercially available biomarker assay kits. Our expertise at serologic assay development was selected by Operation Warp Speed to use the V-PLEX® serology panels as the basis of its standard binding assays for immunogenicity assessments in all funded Phase III clinical trials of COVID vaccines. We will use our MSD MultiArray platform to migrate the top serologic and protein markers for their utility in our target clinical applications. We will collaborate with experts on lung and ovarian cancer screening at Vanderbilt University Medical Center, Boston University, MD Anderson Cancer Center, and German Cancer Research Center, who will also provide access to high-quality well-characterized samples to develop circulating biomarkers to enhance ovarian cancer screening or to distinguish benign from malignant pulmonary nodules. Adhering to the principles of PRoBE design, we will perform Phase I discovery by screening protein arrays with cancer patient and control sera for cancer or control-specific antibodies. Candidate biomarkers for both lung and ovarian cancers will undergo Phase 2 validation.