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

项目总结 生殖细胞肿瘤是青少年和青壮年最常见的实体肿瘤。在切除后 原发肿瘤，许多患者没有其他疾病的证据，被归类为临床肿瘤 I期(CSI)患者。目前对CSI患者的护理标准是对血清肿瘤进行“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 目前可用的标志物AFP和HCG只出现在50%的病例中。一种生物标志物 可以预测术后立即复发的可能性，并准确地检测所有复发 接受监测的患者将使医疗决策合理化，允许立即启动 对复发可能性高的患者进行化疗，并对接受监测的患者更早发现复发。一个 准确的血清生物标志物还将消除对大多数连续监视CT扫描的需要，从而减少 辐射暴露和医疗保健费用。我们的团队已经鉴定了一组4个血清miRNAs(miR-371a-3p， MIR-372-3P、MIR-373-3P和MIR-367-3P)，在迄今回顾分析的1500多个病例中，似乎 无论年龄、性别、原发部位或GCT的主要组织学类型，均普遍升高。我们 已经开发了一套严格的管道来使用定量反向来量化这4个miRNAs的水平 转录聚合酶链式反应。管道中的每一步都最大限度地提高了灵敏度和特异度。在UH2部分期间 在此应用程序中，我们建议进行最后一组必要的实验，以“锁定”分析 过程中，建立这些miRNAs的正常范围，以量化 在案例中的miRNAs，表明这项技术是可转让的，并且一致的结果可以 通过实验室获得，并提出最终建议，进入拨款的UH3部分 敏感性和特异性的临界点。在应用程序的UH3部分，血清miRNA检测将是 在正在进行的临床试验AGCT1531的背景下进行前瞻性评估，该试验目前正在积累 在美国和加拿大的儿科和成人患者，并将于#年在英国开业 2019年。血清miRNAs升高的患病率将在手术后立即确定， 在复发时，对其敏感性、特异性、阴性和阳性预测价值进行前瞻性评估 测试将会进行。根据在这项临床试验期间收集的前瞻性值，最佳 建议使用接收器工作曲线方法确定灵敏度和特异度的临界点。

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.