儿童神经母细胞瘤患者启动子 DNA 甲基化的预后意义。

Prognostic significance of promoter DNA methylation in patients with childhood neuroblastoma.

机构信息

Children's Cancer Institute Australia for Medical Research, Lowy Cancer Research Centre, University of New South Wales, Randwick, NSW, Australia.

出版信息

Clin Cancer Res. 2012 Oct 15;18(20):5690-700. doi: 10.1158/1078-0432.CCR-12-0294. Epub 2012 Aug 28.

DOI:10.1158/1078-0432.CCR-12-0294

PMID:22929802

Abstract

PURPOSE

To characterize the clinical significance of promoter methylation in a cohort of primary neuroblastoma tumors and investigate the association between DNA methylation and clinical outcome.

EXPERIMENTAL DESIGN

A customized Illumina GoldenGate methylation assay was used to assess methylation status of 96 CpG sites within 48 candidate genes in primary neuroblastoma tumors obtained from 131 children diagnosed in Australia. Genes were selected on the basis of previous reports of altered DNA methylation in embryonal cancers. Levels of DNA methylation were validated in a subset of 48 patient samples using combined bisulfite restriction analysis (CoBRA) and bisulfite sequencing. A Cox proportional hazards model was used to investigate the association between promoter hypermethylation and the risk of relapse/death within 5 years of diagnosis, while adjusting for known prognostic factors including MYCN amplification, age, and stage at diagnosis.

RESULTS

Levels of promoter methylation of DNAJC15, neurotrophic tyrosine kinase receptor 1 or TrkA (NTRK1), and tumor necrosis factor receptor superfamily, member 10D (TNFRSF10D), were higher in older patients at diagnosis (P < 0.01), whereas higher levels of methylation of DNAJC15, NTRK1, and PYCARD were observed in patients with MYCN amplification (P < 0.001). In multivariate analysis, hypermethylation of folate hydrolase (FOLH1), myogenic differentiation-1 (MYOD1), and thrombospondin-1 (THBS1) remained significant independent predictors of poorer clinical outcome after adjusting for known prognostic factors (P ≤ 0.017). Moreover, more than 30% of patients displayed hypermethylation in 2 genes or more and were at least 2 times more likely to relapse or die (HR = 2.72, 95% confidence interval = 1.55-4.78, P = 0.001), independent of MYCN status, age, and stage at diagnosis.

CONCLUSIONS

Our findings highlight the potential use of methylation profiling to identify additional prognostic markers and detect new therapeutic targets for selected patient subsets.

摘要

目的

描述原发性神经母细胞瘤肿瘤队列中启动子甲基化的临床意义，并研究 DNA 甲基化与临床结果之间的关联。

实验设计

使用定制的 Illumina GoldenGate 甲基化分析方法，评估从澳大利亚诊断的 131 名儿童的原发性神经母细胞瘤肿瘤中 48 个候选基因的 96 个 CpG 位点的甲基化状态。这些基因是基于胚胎癌中 DNA 甲基化改变的先前报道选择的。使用联合亚硫酸氢盐限制分析 (CoBRA) 和亚硫酸氢盐测序，在 48 个患者样本的亚组中验证了 DNA 甲基化水平。使用 Cox 比例风险模型，在调整已知预后因素（包括 MYCN 扩增、年龄和诊断时的分期）的情况下，研究启动子过度甲基化与诊断后 5 年内复发/死亡风险之间的关联。

结果

在诊断时年龄较大的患者中，DNAJC15、神经营养酪氨酸激酶受体 1 或 TrkA（NTRK1）和肿瘤坏死因子受体超家族成员 10D（TNFRSF10D）的启动子甲基化水平较高（P < 0.01），而在 MYCN 扩增的患者中，DNAJC15、NTRK1 和 PYCARD 的甲基化水平较高（P < 0.001）。在多变量分析中，调整已知预后因素后，叶酸水解酶 (FOLH1)、肌生成分化-1 (MYOD1) 和血小板反应蛋白-1 (THBS1) 的过度甲基化仍然是较差临床结果的独立预测因子（P ≤ 0.017）。此外，超过 30%的患者有 2 个或更多基因的过度甲基化，复发或死亡的可能性至少增加 2 倍（HR = 2.72，95%置信区间 = 1.55-4.78，P = 0.001），与 MYCN 状态、年龄和诊断时的分期无关。