title: Diagnosis of pancreatic lesions collected by endoscopic ultrasound-guided fine-needle aspiration using next-generation sequencing.
abstract: Endoscopic ultrasound-guided fine-needle aspiration (EUF-FNA) has improved the diagnosis of pancreatic lesions. Next-generation sequencing (NGS) facilitates the production of millions of sequences concurrently. Therefore, in the current study, to improve the detectability of oncogenic mutations in pancreatic lesions, an NGS system was used to diagnose EUS-FNA samples. A total of 38 patients with clinically diagnosed EUS-FNA specimens were analyzed; 27 patients had pancreatic ductal adenocarcinoma (PDAC) and 11 had non-PDAC lesions. DNA samples were isolated and sequenced by NGS using an Ion Personal Genome Machine system. The Cancer Hotspot Panel v2, which includes 50 cancer-related genes and 2,790 COSMIC mutations, was used. A >2% mutation frequency was defined as positive.KRASmutations were detected in 26 of 27 PDAC aspirates (96%) and 0 of 11 non-PDAC lesions (0%). The G12, G13, and Q61KRASmutations were found in 25, 0, and 1 of the 27 PDAC samples, respectively. Mutations were confirmed by TaqMan®polymerase chain reaction analysis.TP53mutations were detected in 12 of 27 PDAC aspirates (44%).SMAD4was observed in 3 PDAC lesions and cyclin-dependent kinase inhibitor 2A in 4 PDAC lesions. Therefore, the current study was successfully able to develop an NGS assay with high clinical sensitivity for EUS-FNA samples.
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Eri
authors.initials: E
authors.lastname: Kameta
authors.affiliation: Gastroenterological Centre, Yokohama City University Medical Centre, Yokohama 232-0024, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Sugimori
authors.affiliation: Gastroenterological Centre, Yokohama City University Medical Centre, Yokohama 232-0024, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kaneko
authors.affiliation: Gastroenterological Centre, Yokohama City University Medical Centre, Yokohama 232-0024, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Ishii
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Haruo
authors.initials: H
authors.lastname: Miwa
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sato
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Ishii
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Soichiro
authors.initials: S
authors.lastname: Sue
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Sasaki
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yamashita
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan; Advanced Medical Research Center, Yokohama City University, Yokohama 236-0004, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Shibata
authors.affiliation: Department of Human Genetics, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Naomichi
authors.initials: N
authors.lastname: Matsumoto
authors.affiliation: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Maeda
keywords: KRAS; TP53; endoscopic ultrasound-guided fine-needle aspiration; next-generation sequencing; pancreatic ductal adenocarcinoma