Next‑generation sequencing to identify genetic mutations in pancreatic cancer using intraoperative peritoneal washing fluid

Nakano,Yutaka; Shimane,Gaku; Nakamura,Kohei; Takamatsu,Reika; Aimono,Eriko; Yagi,Hiroshi; Abe,Yuta; Hasegawa,Yasushi; Hori,Shutaro; Tanaka,Masayuki; Masugi,Yohei; Kitago,Minoru; Nishihara,Hiroshi; Kitagawa,Yuko

doi:10.3892/ol.2024.14272

Next‑generation sequencing to identify genetic mutations in pancreatic cancer using intraoperative peritoneal washing fluid

Authors:
- Yutaka Nakano
- Gaku Shimane
- Kohei Nakamura
- Reika Takamatsu
- Eriko Aimono
- Hiroshi Yagi
- Yuta Abe
- Yasushi Hasegawa
- Shutaro Hori
- Masayuki Tanaka
- Yohei Masugi
- Minoru Kitago
- Hiroshi Nishihara
- Yuko Kitagawa
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Affiliations: Department of Surgery, Keio University School of Medicine, Tokyo 160‑8582, Japan, Genomics Unit, Keio University School of Medicine, Tokyo 160‑8582, Japan, Department of Pathology, Keio University School of Medicine, Tokyo 160‑8582, Japan
Published online on: February 2, 2024 https://doi.org/10.3892/ol.2024.14272
Article Number: 139

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Abstract

The efficacy of next‑generation sequencing (NGS) of tumor‑derived DNA from intraoperative peritoneal washing fluid (IPWF) of patients with pancreatic ductal adenocarcinoma (PDAC) who intend to undergo curative resection remains unclear. The aim of the present study was to evaluate whether genomic mutations in tumor‑derived DNA from IPWF samples of patients with PDAC who intend to undergo curative resection could be detected using NGS. A total of 12 such patients were included in this study. Cytology of IPWF (CY) was assessed and NGS of genomic tumor‑derived DNA from the IPWF was performed to determine whether genomic mutations could be detected in these patient samples. A total of 2 patients (16.7%) had a CY(+) status and 1 patient (8.3%) showed intraoperative macro‑peritoneal dissemination; 11 patients underwent radical surgery. Actionable gene alterations were detected in 8 (80.0%) out of the 10 patients with CY(‑) status based on NGS of IPWF samples, and 3 (37.5%) patients among those with actionable gene mutations identified from IPWF samples underwent peritoneal dissemination after surgery within ~12 months. The most common genomic mutation was in KRAS (9 patients, 75.0%), followed by TP53 (3 patients, 25.0%), SMAD4 (1 patient, 8.3%) and CDKN2A (1 patient, 8.3%). These findings indicated that the genomic mutations identified in tumor‑derived DNA from IPWF samples of patients with PDAC with a CY(‑) status who intend to undergo curative resection are potential biomarkers for predicting the recurrence of early peritoneal dissemination.

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1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2020. CA Cancer J Clin. 70:7–30. 2020. View Article : Google Scholar : PubMed/NCBI
2	Ryan DP, Hong TS and Bardeesy N: Pancreatic adenocarcinoma. N Engl J Med. 371:1039–1049. 2014. View Article : Google Scholar : PubMed/NCBI
3	Neoptolemos JP, Kleeff J, Michl P, Costello E, Greenhalf W and Palmer DH: Therapeutic developments in pancreatic cancer: Current and future perspectives. Nat Rev Gastroenterol Hepatol. 15:333–348. 2018. View Article : Google Scholar : PubMed/NCBI
4	Tsuchida H, Fujii T, Mizuma M, Satoi S, Igarashi H, Eguchi H, Kuroki T, Shimizu Y, Tani M, Tanno S, et al: Prognostic importance of peritoneal washing cytology in patients with otherwise resectable pancreatic ductal adenocarcinoma who underwent pancreatectomy: A nationwide, cancer registry-based study from the Japan pancreas society. Surgery. 166:997–1003. 2019. View Article : Google Scholar : PubMed/NCBI
5	Tanaka M, Mihaljevic AL, Probst P, Heckler M, Klaiber U, Heger U, Büchler MW and Hackert T: Meta-analysis of recurrence pattern after resection for pancreatic cancer. Br J Surg. 106:1590–1601. 2019. View Article : Google Scholar : PubMed/NCBI
6	Satoi S, Murakami Y, Motoi F, Uemura K, Kawai M, Kurata M, Sho M, Matsumoto I, Yanagimoto H, Yamamoto T, et al: Reappraisal of peritoneal washing cytology in 984 patients with pancreatic ductal adenocarcinoma who underwent margin-negative resection. J Gastrointest Surg. 19:6–14. 2015. View Article : Google Scholar : PubMed/NCBI
7	Nakano Y, Kitago M, Matsuda S, Nakamura Y, Fujita Y, Imai S, Shinoda M, Yagi H, Abe Y, Hibi T, et al: KRAS mutations in cell-free DNA from preoperative and postoperative sera as a pancreatic cancer marker: a retrospective study. Br J Cancer. 118:662–669. 2018. View Article : Google Scholar : PubMed/NCBI
8	Yokose T, Kitago M, Matsuda S, Sasaki Y, Masugi Y, Nakamura Y, Shinoda M, Yagi H, Abe Y, Oshima G, et al: Combination of KRAS and SMAD4 mutations in formalin-fixed paraffin-embedded tissues as a biomarker for pancreatic cancer. Cancer Sci. 111:2174–2182. 2020. View Article : Google Scholar : PubMed/NCBI
9	Sugimori M, Sugimori K, Tsuchiya H, Suzuki Y, Tsuyuki S, Kaneta Y, Hirotani A, Sanga K, Tozuka Y, Komiyama S, et al: Quantitative monitoring of circulating tumor DNA in patients with advanced pancreatic cancer undergoing chemotherapy. Cancer Sci. 111:266–278. 2020. View Article : Google Scholar : PubMed/NCBI
10	Nagai M, Sho M, Akahori T, Nakagawa K and Nakamura K: Application of liquid biopsy for surgical management of pancreatic cancer. Ann Gastroenterol Surg. 4:216–223. 2020. View Article : Google Scholar : PubMed/NCBI
11	Goldman D and Domschke K: Making sense of deep sequencing. Int J Neuropsychopharmacol. 17:1717–1725. 2014. View Article : Google Scholar : PubMed/NCBI
12	Deveson IW, Gong B, Lai K, LoCoco JS, Richmond TA, Schageman J, Zhang Z, Novoradovskaya N, Willey JC, Jones W, et al: Evaluating the analytical validity of circulating tumor DNA sequencing assays for precision oncology. Nat Biotechnol. 39:1115–1128. 2021. View Article : Google Scholar : PubMed/NCBI
13	Patel A, Hissong E, Rosado L, Burkhardt R, Cong L, Alperstein SA, Siddiqui MT, Parl HJ, Song W, Velu PD, et al: Next-generation sequencing of cell-free DNA extracted from pleural effusion supernatant: Applications and challenges. Front Med (Lausanne). 8:6623122021. View Article : Google Scholar : PubMed/NCBI
14	Chiba K, Hata T, Mizuma M, Matsuda K, Aoki S, Takadate T, Kawaguchi K, Nakagawa K, Morikawa T, Motoi F, et al: Impact of tumor-derived DNA testing in peritoneal lavage of pancreatic cancer patients with and without occult intra-abdominal metastases. Ann Surg Oncol. 29:2685–2697. 2022. View Article : Google Scholar : PubMed/NCBI
15	Yang SR, Mooney KL, Libiran P, Jones CD, Joshi R, Lau HD, Stehr H, Berry GJ, Zehnder JL, Long SR, et al: Targeted deep sequencing of cell-free DNA in serous body cavity fluids with malignant, suspicious, and benign cytology. Cancer Cytopathol. 128:43–56. 2020. View Article : Google Scholar : PubMed/NCBI
16	Bae GE, Kim SH, Choi MK, Kim JM and Yeo MK: Targeted sequencing of ascites and peritoneal washing fluid of patients with gastrointestinal cancers and their clinical applications and limitations. Front Oncol. 11:7127542021. View Article : Google Scholar : PubMed/NCBI
17	Mayo-de-Las-Casas C, Velasco A, Sanchez D, Martínez-Bueno A, Garzón-Ibáñez M, Gatius S, Ruiz-Miró M, Gonzalez-Tallada X, Llordella I, Tresserra F, et al: Detection of somatic mutations in peritoneal lavages and plasma of endometrial cancer patients: A proof-of-concept study. Int J Cancer. 147:277–284. 2020. View Article : Google Scholar : PubMed/NCBI
18	Compton CC and Henson DE: Protocol for the examination of specimens removed from patients with carcinoma of the exocrine pancreas: A basis for checklists. Cancer committee, college of American pathologists. Arch Pathol Lab Med. 121:1129–1136. 1997.PubMed/NCBI
19	Nakamura K, Aimono E, Tanishima S, Imai M, Nagatsuma AK, Hayashi H, Yoshimura Y, Nakayama K, Kyo S and Nishihara Hl: Intratumoral genomic heterogeneity may hinder precision medicine strategies in patients with serous ovarian carcinoma. Diagnostics (Basel). 10:2002020. View Article : Google Scholar : PubMed/NCBI
20	Tsumura K, Arai E, Tian Y, Shibuya A, Nishihara H, Yotani T, Yamada Y, Takahashi Y, Maeshima AM, Fujimoto H, et al: Establishment of permutation for cancer risk estimation in the urothelium based on genome-wide DNA methylation analysis. Carcinogenesis. 40:1308–1319. 2019. View Article : Google Scholar : PubMed/NCBI
21	Shin SH, Kim SC, Hong SM, Kim YH, Song KB, Park KM and Lee YJ: Genetic alterations of K-ras, p53, c-erbB-2, and DPC4 in pancreatic ductal adenocarcinoma and their correlation with patient survival. Pancreas. 42:216–222. 2013. View Article : Google Scholar : PubMed/NCBI
22	Schlitter AM, Segler A, Steiger K, Michalski CW, Jäger C, Konukiewitz B, Pfarr N, Endris V, Bettstetter M, Kong B, et al: Molecular, morphological and survival analysis of 177 resected pancreatic ductal adenocarcinomas (PDACs): Identification of prognostic subtypes. Sci Rep. 7:410642017. View Article : Google Scholar : PubMed/NCBI
23	Olmedillas-López S, García-Arranz M and García-Olmo D: Current and emerging applications of droplet digital PCR in oncology. Mol Diagn Ther. 21:493–510. 2017. View Article : Google Scholar : PubMed/NCBI
24	Huerta M, Roselló S, Sabater L, Ferrer A, Tarazona N, Roda D, Gambardella V, Alfaro-Cervelló C, Garcés-Albir M, Cervantes A and Ibarrola-Villava M: Circulating tumor DNA detection by digital-droplet PCR in pancreatic ductal adenocarcinoma: A systematic review. Cancers (Basel). 13:9942021. View Article : Google Scholar : PubMed/NCBI
25	Yamada S, Fujii T, Yamamoto T, Takami H, Yoshioka I, Yamaki S, Sonohara F, Shibuya K, Motoi F, Hirano S, et al: Phase I/II study of adding intraperitoneal paclitaxel in patients with pancreatic cancer and peritoneal metastasis. Br J Surg. 107:1811–1817. 2020. View Article : Google Scholar : PubMed/NCBI
26	Kilgour E, Rothwell DG, Brady G and Dive C: Liquid biopsy-based biomarkers of treatment response and resistance. Cancer Cell. 37:485–495. 2020. View Article : Google Scholar : PubMed/NCBI
27	Conroy T, Hammel P, Hebbar M, Ben Abdelghani M, Wei AC, Raoul JL, Choné L, Francois E, Artru P, Biagi JJ, et al: FOLFIRINOX or gemcitabine as adjuvant therapy for pancreatic cancer. N Engl J Med. 379:2395–2406. 2018. View Article : Google Scholar : PubMed/NCBI
28	Ueno M, Morinaga S, Hashimoto Y, Umemoto K, Sasahira N, Saiura A, Seyama Y, Honda G, Ioka T, Takahashi H, et al: Tolerability of nab-paclitaxel plus gemcitabine as adjuvant setting in Japanese patients with resected pancreatic cancer: Phase I study. Pancreas. 50:83–88. 2021. View Article : Google Scholar : PubMed/NCBI
29	Samorodnitsky E, Jewell BM, Hagopian R, Miya J, Wing MR, Lyon E, Damodaran S, Bhatt D, Reeser JW, Datta J and Roychowdhury S: Evaluation of hybridization capture versus amplicon-based methods for whole-exome sequencing. Hum Mutat. 36:903–914. 2015. View Article : Google Scholar : PubMed/NCBI
30	Hashimoto D, Arima K, Yokoyama N, Chikamoto A, Taki K, Inoue R, Kaida T, Higashi T, Nitta H, Ohmuraya M, et al: Heterogeneity of KRAS mutations in pancreatic ductal adenocarcinoma. Pancreas. 45:1111–1114. 2016. View Article : Google Scholar : PubMed/NCBI
31	Baldus SE, Schaefer KL, Engers R, Hartleb D, Stoecklein NH and Gabbert HE: Prevalence and heterogeneity of KRAS, BRAF, and PIK3CA mutations in primary colorectal adenocarcinomas and their corresponding metastases. Clin Cancer Res. 16:790–799. 2010. View Article : Google Scholar : PubMed/NCBI
32	Ryan DP, Hong TS and Bardeesy N: Pancreatic adenocarcinoma. N Engl J Med. 371:2140–2141. 2014. View Article : Google Scholar : PubMed/NCBI