Isaka Mitsuhiro, Serizawa Masakuni, Kenmotsu Hirotsugu, Koh Yasuhiro, Takahashi Shoji, Maniwa Tomohiro, Wakuda Kazushige, Ono Akira, Naito Tateaki, Murakami Haruyasu, Mori Keita, Endo Masahiro, Abe Masato, Hayashi Isamu, Nakajima Takashi, Yamamoto Nobuyuki, Takahashi Toshiaki, Ohde Yasuhisa
Division of Thoracic Surgery, Shizuoka Cancer Center, Nagaizumi-cho, Sunto-gun, Japan.
Drug Discovery and Development Division, Research Institute, Shizuoka Cancer Center, Nagaizumi-cho, Sunto-gun, Japan.
Clin Lung Cancer. 2017 Sep;18(5):519-526.e1. doi: 10.1016/j.cllc.2016.11.022. Epub 2016 Dec 2.
Amplicon-based massively parallel sequencing (MPS) is an effective platform for identifying clinically actionable mutations across many genes in limited amounts of tissue. Most lung cancers are diagnosed and staged using small tissue samples obtained by transbronchial biopsy (TBB). To determine whether the mutations in TBB specimens detected by amplicon-based MPS reflect those present in the tumors, we compared the mutational profiles of preoperative TBB specimens and corresponding surgically resected specimens.
Fresh-frozen primary tumor specimens from non-small-cell lung cancer patients (n = 46) obtained preoperatively by TBB and during surgical resection were analyzed. The concordance of mutations detected by amplicon-based MPS in the 2 sample types was investigated, and the allele frequency of the mutations common to both specimens from the same patient was determined.
An initial assessment of DNA quantity revealed that 46% of the TBB specimens (21 of 46) had less than the lower limit for amplicon-based MPS. These 21 TBB specimens were consequently omitted from the analysis. Of the 29 mutations detected in the TBB and/or surgically resected specimens from 25 patients, 23 were present in both samples, for a concordance rate of 79%.
Amplicon-based MPS with TBB specimens approximately reflects clinically relevant tumor mutation profiles. However, the rate of TBB specimens with sufficient DNA quantity for amplicon-based MPS was only around 50%. Therefore, surgically resected specimens have a valuable role in exploratory and comprehensive genomic profiling.
基于扩增子的大规模平行测序（MPS）是在有限组织量中鉴定多个基因临床可操作突变的有效平台。大多数肺癌通过经支气管活检（TBB）获取的小组织样本进行诊断和分期。为了确定基于扩增子的MPS检测到的TBB标本中的突变是否反映肿瘤中存在的突变，我们比较了术前TBB标本和相应手术切除标本的突变谱。
分析了通过TBB术前及手术切除时获得的非小细胞肺癌患者（n = 46）的新鲜冷冻原发性肿瘤标本。研究了基于扩增子的MPS在两种样本类型中检测到的突变的一致性，并确定了同一患者的两个标本中共同存在的突变的等位基因频率。
对DNA量的初步评估显示，46%的TBB标本（46个中的21个）低于基于扩增子的MPS的下限。因此，这21个TBB标本被排除在分析之外。在25例患者的TBB和/或手术切除标本中检测到的29个突变中，23个在两个样本中均存在，一致性率为79%。
基于扩增子的MPS与TBB标本大致反映了临床相关的肿瘤突变谱。然而，DNA量足以进行基于扩增子的MPS的TBB标本比例仅约为50%。因此，手术切除标本在探索性和全面的基因组分析中具有重要作用。
