Establishment of Patient-Derived Pancreatic Cancer Organoids from Endoscopic Ultrasound-Guided Fine-Needle Aspiration Biopsies
1
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
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.
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.
참고문헌 (36)
- Mizrahi JD, "Pancreatic cancer", Lancet, 395, pp.2008~2020 (2020)
- Paulson AS, "Therapeutic advances in pancreatic cancer", Gastroenterology, 144, pp.1316~1326 (2013)
- Cowan RW, "Genetic progression of pancreatic cancer", Cancer J, 20, pp.80~84 (2014)
- Vincent A, "Pancreatic cancer", Lancet, 378, pp.607~620 (2011)
- Conroy T, "FOLFIRINOX versus gemcitabine for metastatic pancreatic cancer", N Engl J Med, 364, pp.1817~1825 (2011)
- Villarroel MC, "Personalizing cancer treatment in the age of global genomic analyses : PALB2 gene mutations and the response to DNA damaging agents in pancreatic cancer", Mol Cancer Ther, 10, pp.3~8 (2011)
- Kim MP, "Generation of orthotopic and heterotopic human pancreatic cancer xenografts in immunodeficient mice", Nat Protoc, 4, pp.1670~1680 (2009)
- Boj SF, "Organoid models of human and mouse ductal pancreatic cancer", Cell, 160, pp.324~338 (2015)
- Kaushik G, "Concise review : current status of three-dimensional organoids as preclinical models", Stem Cells, 36, pp.1329~1340 (2018)
- Sato T, "Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche", Nature, 459, pp.262~265 (2009)
- Sato T, "Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium", Gastroenterology, 141, pp.1762~1772 (2011)
- Barker N, "Lgr5(+ve)stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro", Cell Stem Cell, 6, pp.25~36 (2010)
- Huch M, "Unlimited in vitro expansion of adult bi-potent pancreas progenitors through the Lgr5", R-spondin axis, 2013, pp.32, 2708~2721
- Gao D, "Organoid cultures derived from patients with advanced prostate cancer", Cell, 159, pp.176~187 (2014)
- Huch M, "In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration", Nature, 494, pp.247~250 (2013)
- Karthaus WR, "Identification of multipotent luminal progenitor cells in human prostate organoid cultures", Cell, 159, pp.163~175 (2014)
- Tiriac H, "Successful creation of pancreatic cancer organoids by means of EUS-guided fineneedle biopsy sampling for personalized cancer treatment", Gastrointest Endosc, 87, pp.1474~1480 (2018)
- Morganti AG, "A systematic review of resectability and survival after concurrent chemoradiation in primarily unresectable pancreatic cancer", Ann Surg Oncol, 17, pp.194~205 (2010)
- Eloubeidi MA, "Endoscopic ultrasound-guided fine needle aspiration biopsy of patients with suspected pancreatic cancer : diagnostic accuracy and acute and 30-day complications", Am J Gastroenterol, 98, pp.2663~2668 (2003)
- Lacomb JF, "Single-pass vs 2-pass endoscopic ultrasound-guided fine-needle biopsy sample collection for creation of pancreatic adenocarcinoma organoids", Clin Gastroenterol Hepatol, 19, pp.845~847 (2021)
- Vlachogiannis G, "Patient-derived organoids model treatment response of metastatic gastrointestinal cancers", Science, 359, pp.920~926 (2018)
- Seidlitz T, "Human gastric cancer modelling using organoids", Gut, 68, pp.207~217 (2019)
- Ganesh K, "A rectal cancer organoid platform to study individual responses to chemoradiation", Nat Med, 25, pp.1607~1614 (2019)
- Driehuis E, "Pancreatic cancer organoids recapitulate disease and allow personalized drug screening", Proc Natl Acad Sci U S A, 116, pp.26580~26590 (2019)
- National Institute of Environmental Health Sciences, "Guidance document on using in vitro data to estimate in vivo starting doses for acute toxicity: NIH publication No. 01-4500" (2001), National Toxicology Program
- Stein SM, "Final analysis of a phase II study of modified FOLFIRINOX in locally advanced and metastatic pancreatic cancer", Br J Cancer, 114, pp.737~743 (2016)
- KleeffJ, "Pancreatic cancer", Nat Rev Dis Primers, 2, pp.16022~ (2016)
- Chen JY, "Slow-pull and different conventional suction techniques in endoscopic ultrasoundguided fine-needle aspiration of pancreatic solid lesions using 22-gauge needles", World J Gastroenterol, 22, pp.8790~8797 (2016)
- Huang L, "Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell-and patient-derived tumor organoids", Nat Med, 21, pp.1364~1371 (2015)
- Gendoo DMA, "Whole genomes define concordance of matched primary, xenograft, and organoid models of pancreas cancer", PLoS Comput Biol, 15, pp.e1006596~ (2019)
- Mapes BL, "Universal genetic and transcriptomic concordance metrics to validate patientderived tumor organoid models", Cancer Res, 80(16Suppl), pp.3908~ (2020)
- Tiriac H, "Organoid profiling identifies common responders to chemotherapy in pancreatic cancer", Cancer Discov, 8, pp.1112~1129 (2018)
- Broutier L, "Human primary liver cancer-derived organoid cultures for disease modeling and drug screening", Nat Med, 23, pp.1424~1435 (2017)
- Li X, "Organoid cultures recapitulate esophageal adenocarcinoma heterogeneity providing a model for clonality studies and precision therapeutics", Nat Commun, 9, pp.2983~ (2018)
- Baker LA, "Modeling pancreatic cancer with organoids", Trends Cancer, 2, pp.176~190 (2016)
- Yachida S, "Distant metastasis occurs late during the genetic evolution of pancreatic cancer", Nature, 467, pp.1114~1117 (2010)