Assay Validation of a Circulating miRNA Test for Diagnosis and Monitoring of Malignant Germ Cell Tumors

项目摘要 生殖细胞肿瘤（GCT）是青少年和年轻人最常见的实体肿瘤。切除术后 在原发性肿瘤中，许多患者没有其他疾病的证据，并被归类为“临床 I期（CSI）患者。目前CSI患者的标准治疗是“主动监测”， 标记和成像。然而，这些患者中有20-50%会复发。此外，血清肿瘤 目前可用的标记物AFP和HCG仅存在于50%的病例中。的生物标记物 可以预测术后即刻复发的可能性，并在所有患者中准确检测复发。 接受监测的患者将使医疗决策合理化，允许立即开始治疗。 在复发可能性高的患者中进行化疗，并在监测中更早地发现复发。一个 准确的血清生物标志物也将减少大多数连续监测CT扫描的需要， 辐射暴露和医疗保健费用。我们的小组已经鉴定了一组4种血清miRNA（miR-371 a-3 p， miR-372- 3 p、miR-373- 3 p和miR-367- 3 p），在迄今为止回顾性分析的超过1500例病例中，似乎 无论年龄、性别、原发部位或GCT的主要组织学如何，均普遍升高。我们 我已经开发了一个严格的管道，使用定量逆转录病毒来定量这4种miRNAs的水平。 转录PCR。管道中的每一步都能最大限度地提高灵敏度和特异性。在UH 2部分期间， 在这个应用程序中，我们建议进行最后一组必要的实验，以“锁定”分析 过程中，建立一个正常范围内的这些miRNA，通过它来量化的程度的升高， miRNAS在病例中的应用，证明了这项技术是可转移的，并且可以得到一致的结果。 获得跨实验室，并提出最终建议进入UH 3部分的赠款， 敏感性和特异性的临界点。在申请的UH 3部分中，血清miRNA检测将被 在一项正在进行的临床试验AGCT 1531的背景下进行了前瞻性评价，该试验目前正在进行 美国和加拿大的儿科和成人患者，并将于 2019.将在术后即刻确定血清miRNA升高的患病率， 在复发时，前瞻性评估的敏感性，特异性，阴性和阳性预测值， 将进行测试。根据本临床试验期间前瞻性收集的数值， 使用受试者工作曲线方法，推荐敏感性和特异性的临界点。

PROJECT SUMMARY Germ cell tumors (GCT) are the most common solid tumor of adolescents and young adults. After resection of the primary tumor, many patients do not have evidence of disease elsewhere and are categorized as “clinical stage I (CSI) patients.” The current standard of care for CSI patients is “active surveillance” with serum tumor markers and imaging. However, between 20-50% of these patients will relapse. Moreover, the serum tumor markers that are currently available, AFP and HCG, are present in only 50% of the cases. A biomarker that could predict the likelihood of relapse in the immediate post-op period and detect relapse accurately in all patients on surveillance would rationalize medical decision-making, allowing for immediate initiation of chemotherapy in those at high likelihood of relapse and earlier detection of relapse of those on surveillance. An accurate serum biomarker would also obviate the need for most of the serial surveillance CT scans, reducing radiation exposure and health care costs. Our group has identified a panel of 4 serum miRNAs (miR-371a-3p, miR-372-3p, miR-373-3p and miR-367-3p) that, in over 1500 cases analyzed retrospectively to date, appear to be universally elevated regardless of age, gender, site of primary, or the principal histology of the GCT. We have developed a rigorous pipeline to quantify the levels of these 4 miRNAs using quantitative reverse transcription PCR. Each step in the pipeline maximizes sensitivity and specificity. During the UH2 portion of this application, we propose to conduct the final set of experiments necessary to “lock-down” the analytic process, establish a normal range of these miRNAs by which to quantify the degree of elevation of the miRNAS in cases, demonstrate that this technology is transferrable and that concordant results can be obtained across laboratories, and make final recommendations going into the UH3 portion of the grant on cutpoints for sensitivity and specificity. In the UH3 portion of the application, the serum miRNA test will be prospectively evaluated in the context of an ongoing clinical trial, AGCT1531, that is currently accruing pediatric and adult patients in the United States and Canada and will be opening in the United Kingdom in 2019. The prevalence of the elevated serum miRNAs will be determined in the immediate post-op period and at relapse, and a prospective assessment of the sensitivity, specificity, negative and positive predictive value of the test will be undertaken. With the values gathered prospectively during this clinical trial, the optimal cutpoints for sensitivity and specificity will be recommended, using receiver operating curve methodology.