Noninvasive Diagnosis of Actionable Mutations by Deep Sequencing of Circulating Free DNA in Lung Cancer from Never-Smokers: A Proof-of-Concept Study from BioCAST/IFCT-1002
Noninvasive Diagnosis of Actionable Mutations by Deep Sequencing of Circulating Free DNA in Lung Cancer from Never-Smokers: A Proof-of-Concept Study from BioCAST/IFCT-1002
复制标题
DOI：
10.1158/1078-0432.ccr-13-3063
复制
发表时间：
2014-09-01
影响因子：
11.5
通讯作者：
Souquet, Pierre-Jean
中科院分区：
文献类型：
--
作者：
Couraud, Sebastien;Vaca-Paniagua, Felipe;Souquet, Pierre-Jean
目的：肿瘤体细胞突变分析是转移性肺癌标准管理的一部分。然而，医生往往不得不处理小的活检，因此具有挑战性的突变检测。循环游离DNA（cfDNA）是一种有前途的工具，用于访问肿瘤基因组作为液体活检。在这里，我们评估了下一代测序（NGS）的cfDNA样品从一系列连续的患者筛选的一系列临床相关的mutation.Experimental设计：共107个血浆样本收集自BioCAST/IFCT-1002肺癌研究（从不吸烟者队列）。其中68例获得了与之匹配的肿瘤DNA（tDNA）。设计了基于多重PCR的检测以靶向EGFR、KRAS、BRAF、ERBB 2和PI 3 KCA基因中的特异性编码区，并使用NGS IonTorrent个人基因组机器平台在cfDNA/tDNA对上以深度覆盖进行扩增子测序。在tDNA中，鉴定了50个突变（36个EGFR、5个ERBB 2、4个KRAS、3个BRAF和2个PIK 3CA），其中26个在cfDNA中检测到。该测试的灵敏度为58%（95%置信区间，43%-71%），估计特异性为87%（62%-96%）。结论：这些数据证明了使用IonTorrent NGS在cfDNA中进行突变筛查用于检测转移性肺癌患者中的一系列肿瘤生物标志物的可行性和潜在效用。(C)2014年AACR。
Purpose: Tumor somatic mutation analysis is part of the standard management of metastatic lung cancer. However, physicians often have to deal with small biopsies and consequently with challenging mutation testing. Circulating freeDNA(cfDNA) is a promising tool for accessing the tumor genome as a liquid biopsy. Here, we evaluated next-generation sequencing (NGS) on cfDNA samples obtained from a consecutive series of patients for the screening of a range of clinically relevant mutations.Experimental Design: A total of 107 plasma samples were collected from the BioCAST/IFCT-1002 lung cancer study (never-smokers cohort). Matched tumor DNA (tDNA) was obtained for 68 cases. Multiplex PCR-based assays were designed to target specific coding regions in EGFR, KRAS, BRAF, ERBB2, and PI3KCA genes, and amplicon sequencing was performed at deep coverage on the cfDNA/tDNA pairs using the NGS IonTorrent Personal Genome Machine Platform.Results: CfDNA concentration in plasma was significantly associated with both stage and number of metastatic sites. In tDNA, 50 mutations (36 EGFR, 5 ERBB2, 4 KRAS, 3 BRAF, and 2 PIK3CA) were identified, of which 26 were detected in cfDNA. Sensitivity of the test was 58% (95% confidence interval, 43%-71%) and the estimated specificity was 87% (62%-96%).Conclusion: These data demonstrate the feasibility and potential utility of mutation screening in cfDNA using IonTorrent NGS for the detection of a range of tumor biomarkers in patients with metastatic lung cancer. (C) 2014 AACR.