Molecular Diagnostics for Risk Stratification and Monitoring of Neuroblastoma

项目总结/摘要 尽管在过去，对高危转移性神经母细胞瘤儿童的治疗已经有了显著的改善， 20年来，这些患者中只有45%成为长期的无病生存者，骨髓是一个常见的 耐药或复发性疾病的部位。生存率的提高需要有效的新疗法 基于诊断时基于生物学的风险分层和对 该治疗将识别具有不同结局的患者亚组，这些患者亚组目前在临床上无法区分。 高危神经母细胞瘤很少有复发性体细胞突变，这就排除了分子分类， 依赖于频繁改变的致癌驱动因子的疗法。我们开发了一种新颖的， TaqMan（R）低密度阵列（TLDA）基因表达测定用于诊断时的鉴别和用于 定量该组患者骨髓和血液中的"肿瘤负荷"。总体目标：提高临床 与儿童肿瘤学小组（COG）合作，利用这些新的分子诊断方法， 将其确立为CAP/CLIA认证的测试。具体目的：确认性能特征和临床 未处理的原发性MYCN非扩增的预测特征（14-基因TLDA测定）的效用1） 2）用于定量骨中"肿瘤负荷"的检测标记（5-基因TLDA测定） 骨髓和血液。3)确定这些检测单独或联合是否可改善对患者结局的预测。 影像学和临床变量的背景。研究设计和方法：性能特征 TLDA检测用于神经母细胞瘤风险分层和反应监测的临床效用将是 经过验证，并将在CAP/CLIA认证的实验室（包括儿童医院的实验室）进行检测 洛杉矶[CHLA]和全国儿童/COG生物病理学中心[BPC]，以便它们可以用于 临床决策的未来。这些检测的分析性能特征将在 CHLA和BPC实验室，临床效用研究将在CHLA进行。14基因预测 在三个独立的临床试验中，签名预测了结果，我们相信，更多的研究， 使用来自四项COG研究的300份存档和前瞻性样本将复制现有数据。初步研究 5个基因检测标记（CHGA、DCX、DDC、PHOX 2B和TH），其可以识别一个肿瘤细胞 106例正常骨髓或血细胞中的肿瘤负荷显示， 诱导、清髓性巩固治疗后和巩固治疗后与结果相关。 将使用2，250份存档和前瞻性骨髓、血液和PBSC标本对这些结果进行验证 四项COG研究。旨在优化临床分层和监测的综合分析 决策将包括这些分子诊断测试，131 I-间碘苄胍（MIBG）成像， 临床变量。总结：我们预期我们的分子诊断试验将提高对 结果和发展更有效的治疗高危转移性神经母细胞瘤患者。

Project Summary/Abstract Although treatment for children with high-risk metastatic neuroblastoma has improved significantly in the past 20 years, only 45% of these patients become long-term, disease-free survivors, and bone marrow is a frequent site of resistant or recurrent disease. Improvement in survival necessitates effective new therapies for patients based upon biologically based risk stratification at diagnosis and highly sensitive monitoring of response to therapy that will identify patient subgroups with different outcomes who currently are clinically indistinguishable. High-risk neuroblastomas have few recurrent somatic mutations, which obviates molecular classification and therapies that rely on frequently altered oncogenic drivers. We have developed novel and clinically applicable TaqMan(R) Low Density Array (TLDA) gene expression assays for prognostication at diagnosis and for quantifying "tumor load" in bone marrow and blood for this group of patients. Overall goal: Validate the clinical utility of these new molecular diagnostics in collaboration with the Children's Oncology Group (COG) and then establish them as CAP/CLIA-accredited tests. Specific Aims: Confirm performance characteristics and clinical utility 1) of a prediction signature (14-gene TLDA assay) for untreated primary MYCN non-amplified neuroblastomas and 2) of a detection signature (5-gene TLDA assay) for quantifying "tumor load" in bone marrow and blood. 3) Determine if these assays alone or combined improve prediction of outcome in the context of imaging and clinical variables. Research Design and Methods: The performance characteristics and clinical utility of the TLDA assays for risk stratification and monitoring of neuroblastoma response will be validated, and tests will be installed in CAP/CLIA-accredited laboratories including those at Children's Hospital Los Angeles [CHLA] and Nationwide Children's/COG Biopathology Center [BPC] so that they may be used in the future for clinical decision-making. Analytical performance characteristics of these tests will be confirmed in both CHLA and BPC laboratories, and clinical utility studies will be performed at CHLA. The 14-gene prediction signature predicted outcome in three independent clinical trials, and we are confident that additional studies using 300 archived and prospective samples from four COG studies will replicate existing data. Initial studies of the 5-gene detection signature (CHGA, DCX, DDC, PHOX2B, and TH), which can identify one tumor cell among 106 normal bone marrow or blood cells, showed that tumor load in bone marrow and PBSC during induction, after myeloablative consolidation, and after post-consolidation therapy correlates with outcome. These results will be validated with 2,250 archived and prospective bone marrow, blood, and PBSC specimens from four COG studies. Comprehensive analyses aimed at optimizing stratification and monitoring for clinical decision making will include these molecular diagnostic tests, 131I-meta-iodobenzyguanidine (MIBG) imaging, and clinical variables. Summary: We anticipate that our moelcular diagnostic assays will improve prediction of outcome and development of more effective therapy for patients with high-risk metastatic neuroblastoma.