Gut Liver. 2022;16(4):625-636.

Establishment of Patient-Derived Pancreatic Cancer Organoids from Endoscopic Ultrasound-Guided Fine-Needle Aspiration Biopsies.

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제1저자 이지형, 김혜령 · 교신저자 이상협, 구자록

PMID 34916338 PMC9289822 DOI 10.5009/gnl210166 IF 4.1 소화기내과

소속 (6)

Department of Internal Medicine and Liver Research Institute, Seoul National University Hospital, Seoul National University College of Medicine, Seoul, Korea.
Natural Products Research Institute, Seoul National University College of Pharmacy, Seoul, Korea.
Department of Pathology, Seoul National University Hospital, Seoul National University College of Medicine, Seoul, Korea.
Department of Biomedical Sciences, Korean Cell Line Bank, Laboratory of Cell Biology and Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
Center for Liver and Pancreatobiliary Cancer, National Cancer Center, Goyang, Korea.
Division of Surgical Oncology, Department of Surgery, Moores Cancer Center, University of California, San Diego, La Jolla, CA, USA.

Abstract

BACKGROUND/AIMS Three-dimensional cultures of human pancreatic cancer tissue also known as "organoids" have largely been developed from surgical specimens. Given that most patients present with locally advanced and/or metastatic disease, such organoids are not representative of the majority of patients. Therefore, we used endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) to collect pancreatic cancer tissues from patients with advanced pancreatic cancer to create organoids, and evaluated their utility in pancreatic cancer research.

METHODS Single-pass EUS-FNA samplings were employed to obtain the tissue for organoid generation. After establishment of the organoid, we compared the core biopsy tissues with organoids using hematoxylin and eosin staining, and performed whole exome sequencing (WES) to detect mutational variants. Furthermore, we compared patient outcome with the organoid drug response to determine the potential utility of the clinical application of such organoid-based assays.

RESULTS Organoids were successfully generated in 14 of 20 tumors (70%) and were able to be passaged greater than 5 times in 12 of 20 tumors (60%). Among them, we selected eight pairs of organoid and core biopsy tissues for detailed analyses. They showed similar patterns in hematoxylin and eosin staining. WES revealed mutations in KRAS, TP53, CDKN2A, SMAD4, BRCA1, and BRCA2 which were 93% homologous, and the mean nonreference discordance rate was 5.47%. We observed moderate drug response correlations between the organoids and clinical outcomes in patients who underwent FOLFIRINOX chemotherapy.

CONCLUSIONS The established organoids from EUS-FNA core biopsies can be used for a suitable model system for pancreatic cancer research.

Keywords

Endoscopic ultrasound-guided fine-needle aspirationFOLFIRINOXHigh-throughput nucleotide sequencingOrganoidPancreatic neoplasms

Figures

**Fig. 1** Study scheme. Among 20 biopsy tissues, 12 organoids were established. Histological & genetic comparisons (eight organoids); SNU-3947-TO, SNU-4158-TO, SNU-4208-TO, SNU-4309-TO, SNU-4354-TO, SNU-4365-TO, SNU-4425-TO, and SNU-4607-TO. Correlation analysis between drug response and clinical data (eight organoids); SNU-4158-TO, SNU-4192-TO, SNU-4206-TO, SNU-4208-TO, SNU-4305-TO, SNU-4354-TO, SNU-4365-TO, and SNU-4607-TO.

**Fig. 2** (A) Organoid generation from endoscopic ultrasound-guided fine-needle biopsy sampling. (B) Bright-field microscopy images of 12 established organoids. Scale bars, 400 µm. EUS-FNA, endoscopic ultrasound-guided fine-needle aspiration; DPBS, Dulbecco's phosphate-buffered saline.

**Fig. 3** Representative H&E staining of patient biopsy tissues and pancreatic cancer-derived organoids and corresponding bright-field microscopy images. Scale bars: 400 µm.

**Fig. 4** (A) Venn diagrams of mutations present in biopsy tissues and organoids. (B) Mutational comparison of organoids and biopsy tissues. Representative mutations: *KRAS, TP53, CDKN2A, BRCA1, BRCA2*, and *ARID1A* in an organoid and biopsy tissue pair (SNU3947-TO, organoid; SNU-3947-tissue, biopsy tissue).

**Fig. 5** (A) X-axis, overall survival months of patients who underwent the FOLFIRINOX regimen; Y-axis, overall survival of organoids (SNU-4365-PO, SNU-4365-patient overall survival); the AUCs of eight organoids treated with the FOLFIRINOX regimen are illustrated by heatmap (0<AUC<1). (B) Correlation between patient overall survival and AUC by Spearman’s correlation coefficient (rs). FOLFIRINOX, oxaliplatin, 5-FU, irinotecan, and leucovorin; AUC, area under the curve.

그림 출처: PubMed Central 공개 논문 (PMC9289822)

Conflict of interest statement

CONFLICTS OF INTEREST . J.K.R. is an editorial board member of the journal but was not involved in the peer reviewer selection, evaluation, or decision process of this article. No other potential conflicts of interest relevant to this article were reported.

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