DNA methylation biomarkers offer improved diagnostic efficiency in lung cancer.
DNA methylation biomarkers offer improved diagnostic efficiency in lung cancer.
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DOI：
10.1158/0008-5472.can-12-2309
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发表时间：
2012-11-15
期刊：
影响因子：
11.2
通讯作者：
Liloglou T
中科院分区：
文献类型：
--
作者：
Nikolaidis G;Raji OY;Markopoulou S;Gosney JR;Bryan J;Warburton C;Walshaw M;Sheard J;Field JK;Liloglou T
肺癌异常高的死亡率在很大程度上是由晚期诊断引起的。尽管有大量关于潜在的肺癌诊断分子生物标志物的研究，但很少有临床应用。在这项研究中，我们开发了一组DNA甲基化生物标志物，并通过大型回顾性队列验证了它们在支气管洗涤中的诊断效率。先前高通量方法的候选靶点通过焦磷酸测序在48对独立的肺肿瘤/正常配对中进行检测。选择10个启动子，开发定量甲基化特异性PCR （qMSP）检测方法，用于筛选来自利物浦肺项目（LLP）受试者的655例支气管洗涤（BWs），分为训练组（194例和214例对照）和验证组（139例和109例对照）。采用三种统计模型来选择最优的标记组，并对区分算法的性能进行评价。最终的logit回归模型纳入了p16、TERT、WT1和RASSF1位点的超甲基化。该4基因甲基化特征在验证集中的表现为82%的敏感性和91%的特异性。相比之下，单独的细胞学检查提供43%的敏感性和100%的特异性。该小组的诊断效率没有显示出年龄、性别、吸烟和非肺肿瘤存在的任何偏差。然而，中央（鳞状和小细胞）肿瘤的敏感性可预测高于周围（腺癌）肿瘤，以及2期或更大的肿瘤。这些发现清楚地证明了基于DNA甲基化的检测在细胞学上隐匿性肺肿瘤诊断中的作用。目前，LLP研究即将进行一项前瞻性试验，以提供临床环境中提高诊断准确性的数据，包括通过其他标记物。
The exceptional high mortality of lung cancer can be instigated to a high degree by late diagnosis. Despite the plethora of studies on potential molecular biomarkers for lung cancer diagnosis, very few have reached clinical implementation. In this study we developed a panel of DNA methylation biomarkers and validated their diagnostic efficiency in bronchial washings from a large retrospective cohort. Candidate targets from previous high-throughput approaches were examined by Pyrosequencing in an independent set of 48 lung tumor/normal paired. Ten promoters were selected and quantitative methylation-specific PCR (qMSP) assays were developed and used to screen 655 bronchial washings (BWs) from the Liverpool Lung Project (LLP) subjects divided into training (194 cases and 214 Controls) and validation (139 cases and 109 controls) sets. Three statistical models were employed to select the optimal panel of markers and evaluate the performance of the discriminatory algorithms. The final logit regression model incorporated hypermethylation at p16, TERT, WT1 and RASSF1.The performance of this 4-gene methylation signature in the validation set demonstrated 82% sensitivity and 91% specificity. In comparison, cytology alone in this set provided 43% sensitivity at 100% specificity. The diagnostic efficiency of the panel did not show any biases with age, gender, smoking and the presence of a non-lung neoplasm. However, sensitivity was predictably higher in central (squamous and small cell) than peripheral (adenocarcinomas) tumors, as well as in stage 2 or greater tumors.These findings clearly demonstrate the impact of DNA methylation-based assays in the diagnosis of cytologically occult lung neoplasms. A prospective trial is currently imminent in the LLP study to provide data on the enhancement of diagnostic accuracy in a clinical setting, including by additional markers.