A Novel Approach for the Genetic Analysis of Biliary Tract Cancer Specimens Obtained Through Endoscopic Ultrasound-Guided Fine Needle Aspiration Using Targeted Amplicon Sequencing
A Novel Approach for the Genetic Analysis of Biliary Tract Cancer Specimens Obtained Through Endoscopic Ultrasound-Guided Fine Needle Aspiration Using Targeted Amplicon Sequencing
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DOI：
10.14309/ctg.0000000000000022
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发表时间：
2019-03-19
影响因子：
3.6
通讯作者：
Sakamoto, Naoya
中科院分区：
文献类型：
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作者：
Hirata, Koji;Kuwatani, Masaki;Sakamoto, Naoya
目的：胆道癌（BTC）是一种侵袭性恶性肿瘤，基于生物标志物的临床试验目前正在进行中。内镜超声引导下的细针穿刺（EUS-FNA）是一种安全的手术，可以进行病理诊断；然而，通过EUS-FNA获得的微小BTC肿瘤样本是否可以分析BTC发展和耐受性中的各种遗传改变尚不确定。因此，我们的目的是验证用BTC的EUS-FNA样本进行遗传分析的可行性。方法：使用包含50个基因的癌症基因面板对21例BTC患者的组织样本进行靶向扩增子测序，这些组织样本通过EUS-FNA获得，采用一种新的快速现场处理方法与配对的外周血样本进行比较。结果：21例BTC EUS-FNA标本中有20例（95.2%）成功鉴定出致病基因改变，其中19例为腺癌，2例为腺鳞癌。共鉴定出39个基因的80个单核苷酸变异和8个indel，鉴定出14个基因的28个致病改变（平均每例1.4个改变）。胆囊癌中最常见的改变是TP53、KRAS和CDKN2A；TP53、KRAS、PIK3CA和BRAF在肝内胆管癌中的作用TP53和smad4在肝外胆管癌中的作用。21例患者中有7例发现可操作基因改变（BRAF、NRAS、PIK3CA和IDH1）。结论：利用EUS-FNA获得的BTC标本进行靶向扩增子测序进行遗传分析的新方法是可行的，并使我们能够识别基因组改变。
OBJECTIVES: Biliary tract cancer (BTC) is an aggressive malignant tumor, and biomarker-based clinical trials for this cancer are currently ongoing. Endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) is a safe procedure and enables pathological diagnoses; however, it is uncertain whether a tiny tumor sample of BTC obtained through EUS-FNA can be analyzed for diverse genetic alterations in the development and tolerance of BTC. Thus, we aimed to verify the feasibility of genetic analyses with EUS-FNA samples of BTC.METHODS: Targeted amplicon sequencing using a cancer gene panel with 50 genes was performed with tissue samples of 21 BTC patients obtained through EUS-FNA with a novel rapid on-site process compared with paired peripheral blood samples.RESULTS: Pathogenic gene alterations were successfully identified in 20 out of 21 patients (95.2%) with EUS-FNA specimens of BTC, which included 19 adenocarcinomas and 2 adenosquamous carcinomas. Eighty single nucleotide variants and 8 indels in 39 genes were identified in total, and 28 pathogenic alterations in 14 genes were identified (average, 1.4 alterations per patient). The most common alterations were TP53, KRAS, and CDKN2A in gallbladder carcinoma; TP53, KRAS, PIK3CA, and BRAF in intrahepatic cholangiocarcinoma; and TP53 andSMAD4 in extrahepatic cholangiocarcinoma. Actionable gene alterations (BRAF, NRAS, PIK3CA, and IDH1) were identified in 7 out of 21 patients.CONCLUSIONS: A novel approach in genetic analysis using targeted amplicon sequencing with BTC specimens obtained through EUS-FNA was feasible and enabled us to identify genomic alterations.