Identification of novel mutations in FFPE lung adenocarcinomas using DEPArray sorting technology and next-generation sequencing.
Identification of novel mutations in FFPE lung adenocarcinomas using DEPArray sorting technology and next-generation sequencing.
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DOI：
10.1007/s13353-018-0439-4
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发表时间：
2018-08
影响因子：
2.4
通讯作者：
Seo JS
中科院分区：
文献类型：
--
作者：
Lee JW;Shin JY;Seo JS
福尔马林固定石蜡包埋 (FFPE) 组织被用作病理实验室的标准诊断方法。然而，不需要的组织的混合和可用于检测体细胞突变的正常样本的短缺被认为是准确诊断癌症的关键因素。为了探索这些挑战，我们通过双电泳阵列 (DEPArray) 技术（一种新的细胞分选技术）对 22 个 FFPE 肺腺癌组织中的纯肿瘤细胞进行分选，并使用新一代测序 (NGS) 分析变异，以进行最准确的分析。在通过DEPArray分选的细胞中，所有基因突变的等位基因频率提高了1.2倍（抑癌基因，1.3-10.1倍；癌基因，1.3-2.6倍）。我们通过 DEPArray 技术对分选细胞进行测序，鉴定出 16 个新突变，而通过对未分选细胞进行测序，检测到 4 个新突变。通过此分析，我们还发现五个基因（TP53、EGFR、PTEN、RB1、KRAS 和 CTNNB1）在多个同质肺腺癌中发生体细胞突变。我们一起通过 DEPArray 技术从 22 个 FFPE 肺腺癌中分选纯肿瘤细胞，并鉴定出 16 个新的体细胞突变。我们还建立了精确的基因组图谱，以便更准确地诊断 22 种肺腺癌，并在纯肿瘤细胞中检测到突变。这项研究获得的结果可以为鳞状细胞肺癌的治疗和诊断提供新的途径。本文的在线版本 (10.1007/s13353-018-0439-4) 包含补充材料，可供授权用户使用。
Formalin-fixed paraffin-embedded (FFPE) tissues are utilized as the standard diagnostic method in pathology laboratories. However, admixture of unwanted tissues and shortage of normal samples, which can be used to detect somatic mutation, are considered critical factors to accurately diagnose cancer. To explore these challenges, we sorted the pure tumor cells from 22 FFPE lung adenocarcinoma tissues via Di-Electro-Phoretic Array (DEPArray) technology, a new cell sorting technology, and analyzed the variants with next-generation sequencing (NGS) for the most accurate analysis. The allele frequencies of the all gene mutations were improved by 1.2 times in cells sorted via DEPArray (tumor suppressor genes, 1.3–10.1 times; oncogenes, 1.3–2.6 times). We identified 16 novel mutations using the sequencing from sorted cells via DEPArray technology, compared to detecting 4 novel mutation by the sequencing from unsorted cells. Using this analysis, we also revealed that five genes (TP53, EGFR, PTEN, RB1, KRAS, and CTNNB1) were somatically mutated in multiple homogeneous lung adenocarcinomas. Together, we sorted pure tumor cells from 22 FFPE lung adenocarcinomas by DEPArray technology and identified 16 novel somatic mutations. We also established the precise genomic landscape for more accurate diagnosis in 22 lung adenocarcinomas with mutations detected in pure tumor cells. The results obtained in this study could offer new avenues for the treatment and the diagnosis of squamous cell lung cancers. The online version of this article (10.1007/s13353-018-0439-4) contains supplementary material, which is available to authorized users.
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影响因子：
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作者：
Thorvaldsdóttir H;Robinson JT;Mesirov JP
通讯作者：
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通讯作者：
Piver, MS
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通讯作者：
Scarpa A
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4.6
作者：
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通讯作者：
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