Diagnosis of pancreatic lesions collected by endoscopic ultrasound-guided fine-needle aspiration using next-generation sequencing

Kameta,Eri; Sugimori,Kazuya; Kaneko,Takashi; Ishii,Tomohiro; Miwa,Haruo; Sato,Takeshi; Ishii,Yasuaki; Sue,Soichiro; Sasaki,Tomohiko; Yamashita,Yuki; Shibata,Wataru; Matsumoto,Naomichi; Maeda,Shin

doi:10.3892/ol.2016.5168

Diagnosis of pancreatic lesions collected by endoscopic ultrasound-guided fine-needle aspiration using next-generation sequencing

Authors:
- Eri Kameta
- Kazuya Sugimori
- Takashi Kaneko
- Tomohiro Ishii
- Haruo Miwa
- Takeshi Sato
- Yasuaki Ishii
- Soichiro Sue
- Tomohiko Sasaki
- Yuki Yamashita
- Wataru Shibata
- Naomichi Matsumoto
- Shin Maeda
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Affiliations: Department of Gastroenterology, Yokohama City University Graduate School of Medicine, Yokohama 236‑0004, Japan, Gastroenterological Centre, Yokohama City University Medical Centre, Yokohama 232‑0024, Japan, Department of Human Genetics, Yokohama City University Graduate School of Medicine, Yokohama 236‑0004, Japan
Published online on: September 21, 2016 https://doi.org/10.3892/ol.2016.5168
Pages: 3875-3881
Copyright: © Kameta et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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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. KRAS mutations were detected in 26 of 27 PDAC aspirates (96%) and 0 of 11 non‑PDAC lesions (0%). The G12, G13, and Q61 KRAS mutations were found in 25, 0, and 1 of the 27 PDAC samples, respectively. Mutations were confirmed by TaqMan® polymerase chain reaction analysis. TP53 mutations were detected in 12 of 27 PDAC aspirates (44%). SMAD4 was 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.

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1	Vincent A, Herman J, Schulick R, Hruban RH and Goggins M: Pancreatic cancer. Lancet. 378:607–620. 2011. View Article : Google Scholar : PubMed/NCBI
2	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2012. View Article : Google Scholar
3	Hamada T, Yasunaga H, Nakai Y, Isayama H, Horiguchi H, Matsuda S, Fushimi K and Koike K: Severe bleeding and perforation are rare complications of endoscopic ultrasound-guided fine needle aspiration for pancreatic masses: An analysis of 3,090 patients from 212 hospitals. Gut Liver. 8:215–218. 2014. View Article : Google Scholar : PubMed/NCBI
4	Katanuma A, Maguchi H, Yane K, Hashigo S, Kin T, Kaneko M, Kato S, Kato R, Harada R, Osanai M, et al: Factors predictive of adverse events associated with endoscopic ultrasound-guided fine needle aspiration of pancreatic solid lesions. Dig Dis Sci. 58:2093–2099. 2013. View Article : Google Scholar : PubMed/NCBI
5	Kato K, Kamada H, Fujimori T, Aritomo Y, Ono M and Masaki T: Molecular biologic approach to the diagnosis of pancreatic carcinoma using specimens obtained by EUS-guided fine needle aspiration. Gastroenterol Res Pract. 2012:2435242012. View Article : Google Scholar : PubMed/NCBI
6	Gillis A, Cipollone I, Cousins G and Conlon K: Does EUS-FNA molecular analysis carry additional value when compared to cytology in the diagnosis of pancreatic cystic neoplasm? A systematic review. HPB (Oxford). 17:377–386. 2015. View Article : Google Scholar : PubMed/NCBI
7	de Biase D, Visani M, Baccarini P, Polifemo AM, Maimone A, Fornelli A, Giuliani A, Zanini N, Fabbri C, Pession A and Tallini G: Next generation sequencing improves the accuracy of KRAS mutation analysis in endoscopic ultrasound fine needle aspiration pancreatic lesions. PLoS One. 9:e876512014. View Article : Google Scholar : PubMed/NCBI
8	Kubota Y, Kawakami H, Natsuizaka M, Kawakubo K, Marukawa K, Kudo T, Abe Y, Kubo K, Kuwatani M, Hatanaka Y, et al: CTNNB1 mutational analysis of solid-pseudopapillary neoplasms of the pancreas using endoscopic ultrasound-guided fine-needle aspiration and next-generation deep sequencing. J Gastroenterol. 50:203–210. 2015. View Article : Google Scholar : PubMed/NCBI
9	Bryant KL, Mancias JD, Kimmelman AC and Der CJ: KRAS: Feeding pancreatic cancer proliferation. Trends Biochem Sci. 39:91–100. 2014. View Article : Google Scholar : PubMed/NCBI
10	Biankin AV, Waddell N, Kassahn KS, Gingras MC, Muthuswamy LB, Johns AL, Miller DK, Wilson PJ, Patch AM, Wu J, et al: Pancreatic cancer genomes reveal aberrations in axon guidance pathway genes. Nature. 491:399–405. 2012. View Article : Google Scholar : PubMed/NCBI
11	Collins MA, Bednar F, Zhang Y, Brisset JC, Galbán S, Galbán CJ, Rakshit S, Flannagan KS, Adsay NV and di Magliano M Pasca: Oncogenic Kras is required for both the initiation and maintenance of pancreatic cancer in mice. J Clin Invest. 122:639–653. 2012. View Article : Google Scholar : PubMed/NCBI
12	Ryan DP, Hong TS and Bardeesy N: Pancreatic adenocarcinoma. N Engl J Med. 371:1039–1049. 2014. View Article : Google Scholar : PubMed/NCBI
13	Jeong J, Park YN, Park JS, Yoon DS, Chi HS and Kim BR: Clinical significance of p16 protein expression loss and aberrant p53 protein expression in pancreatic cancer. Yonsei Med J. 46:519–525. 2005. View Article : Google Scholar : PubMed/NCBI
14	Bardeesy N, Cheng KH, Berger JH, Chu GC, Pahler J, Olson P, Hezel AF, Horner J, Lauwers GY, Hanahan D and DePinho RA: Smad4 is dispensable for normal pancreas development yet critical in progression and tumor biology of pancreas cancer. Genes Dev. 20:3130–3146. 2006. View Article : Google Scholar : PubMed/NCBI
15	Waddell N, Pajic M, Patch AM, Chang DK, Kassahn KS, Bailey P, Johns AL, Miller D, Nones K, Quek K, et al: Whole genomes redefine the mutational landscape of pancreatic cancer. Nature. 518:495–501. 2015. View Article : Google Scholar : PubMed/NCBI
16	Young G, Wang K, He J, Otto G, Hawryluk M, Zwirco Z, Brennan T, Nahas M, Donahue A, Yelensky R, et al: Clinical next-generation sequencing successfully applied to fine-needle aspirations of pulmonary and pancreatic neoplasms. Cancer Cytopathol. 121:688–694. 2013. View Article : Google Scholar : PubMed/NCBI
17	Liang WS, Craig DW, Carpten J, Borad MJ, Demeure MJ, Weiss GJ, Izatt T, Sinari S, Christoforides A, Aldrich J, et al: Genome-wide characterization of pancreatic adenocarcinoma patients using next generation sequencing. PLoS One. 7:e431922012. View Article : Google Scholar : PubMed/NCBI
18	Ab Mutalib NS, Syafruddin SE, Md Zain RR, Dali AZ Mohd, Yunos RI Mohd, Saidin S, Jamal R and Mokhtar NM: Molecular characterization of serous ovarian carcinoma using a multigene next generation sequencing cancer panel approach. BMC Res Notes. 7:8052014. View Article : Google Scholar : PubMed/NCBI
19	Rothé F, Laes JF, Lambrechts D, Smeets D, Vincent D, Maetens M, Fumagalli D, Michiels S, Drisis S, Moerman C, et al: Plasma circulating tumor DNA as an alternative to metastatic biopsies for mutational analysis in breast cancer. Ann Oncol. 25:1959–1965. 2014. View Article : Google Scholar : PubMed/NCBI
20	Cao YY, Qu YJ, Song F, Zhang T, Bai JL, Jin YW and Wang H: Fast clinical molecular diagnosis of hyperphenylalaninemia using next-generation sequencing-based on a custom AmpliSeq™ panel and Ion Torrent PGM sequencing. Mol Genet Metab. 113:261–266. 2014. View Article : Google Scholar : PubMed/NCBI
21	Gonzalez-Perez A, Deu-Pons J and Lopez-Bigas N: Improving the prediction of the functional impact of cancer mutations by baseline tolerance transformation. Genome Med. 4:892012. View Article : Google Scholar : PubMed/NCBI
22	Bournet B, Selves J, Grand D, Danjoux M, Hanoun N, Cordelier P and Buscail L: Endoscopic ultrasound-guided fine-needle aspiration biopsy coupled with a KRAS mutation assay using allelic discrimination improves the diagnosis of pancreatic cancer. J Clin Gastroenterol. 49:50–56. 2015. View Article : Google Scholar : PubMed/NCBI
23	Liu QY, Chang MN, Lei JX, Koukiekolo R, Smith B, Zhang D and Ghribi O: Identification of microRNAs involved in Alzheimer's progression using a rabbit model of the disease. Am J Neurodegener Dis. 3:33–44. 2014.PubMed/NCBI
24	Ramesh J and Varadarajulu S: How can we get the best results with endoscopic ultrasound-guided fine needle aspiration? Clin Endosc. 45:132–137. 2012. View Article : Google Scholar : PubMed/NCBI
25	Lunardi S, Muschel RJ and Brunner TB: The stromal compartments in pancreatic cancer: Are there any therapeutic targets? Cancer Lett. 343:147–155. 2014. View Article : Google Scholar : PubMed/NCBI
26	Fuccio L, Hassan C, Laterza L, Correale L, Pagano N, Bocus P, Fabbri C, Maimone A, Cennamo V, Repici A, et al: The role of K-ras gene mutation analysis in EUS-guided FNA cytology specimens for the differential diagnosis of pancreatic solid masses: A meta-analysis of prospective studies. Gastrointest Endosc. 78:596–608. 2013. View Article : Google Scholar : PubMed/NCBI
27	Yachida S, White CM, Naito Y, Zhong Y, Brosnan JA, Macgregor-Das AM, Morgan RA, Saunders T, Laheru DA, Herman JM, et al: Clinical significance of the genetic landscape of pancreatic cancer and implications for identification of potential long-term survivors. Clin Cancer Res. 18:6339–6347. 2012. View Article : Google Scholar : PubMed/NCBI
28	Blackford A, Serrano OK, Wolfgang CL, Parmigiani G, Jones S, Zhang X, Parsons DW, Lin JC, Leary RJ, Eshleman JR, et al: SMAD4 gene mutations are associated with poor prognosis in pancreatic cancer. Clin Cancer Res. 15:4674–4679. 2009. View Article : Google Scholar : PubMed/NCBI
29	Navas C, Hernández-Porras I, Schuhmacher AJ, Sibilia M, Guerra C and Barbacid M: EGF receptor signaling is essential for k-ras oncogene-driven pancreatic ductal adenocarcinoma. Cancer Cell. 22:318–330. 2012. View Article : Google Scholar : PubMed/NCBI
30	Jancík S, Drábek J, Radzioch D and Hajdúch M: Clinical relevance of KRAS in human cancers. J Biomed Biotechnol. 2010:1509602010. View Article : Google Scholar : PubMed/NCBI