There are few in vitro models of exocrine pancreas development and primary human pancreatic adenocarcinoma (PDAC). We establish three-dimensional culture conditions to induce the differentiation of human pluripotent stem cells into exocrine progenitor organoids that form ductal and acinar structures in culture and in vivo. Expression of mutant KRAS or TP53 in progenitor organoids induces mutation-specific phenotypes in culture and in vivo. Expression of TP53R175H induces cytosolic SOX9 localization. In patient tumors bearing TP53 mutations, SOX9 was cytoplasmic and associated with mortality. We also define culture conditions for clonal generation of tumor organoids from freshly resected PDAC. Tumor organoids maintain the differentiation status, histoarchitecture and phenotypic heterogeneity of the primary tumor and retain patient-specific physiological changes, including hypoxia, oxygen consumption, epigenetic marks and differences in sensitivity to inhibition of the histone methyltransferase EZH2. Thus, pancreatic progenitor organoids and tumor organoids can be used to model PDAC and for drug screening to identify precision therapy strategies.
Subscribe to Journal
Get full journal access for 1 year
only $18.75 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Ghaneh, P., Costello, E. & Neoptolemos, J.P. Biology and management of pancreatic cancer. Postgrad. Med. J. 84, 478–497 (2008).
Kanji, Z.S. & Gallinger, S. Diagnosis and management of pancreatic cancer. CMAJ 185, 1219–1226 (2013).
Vincent, A., Herman, J., Schulick, R., Hruban, R.H. & Goggins, M. Pancreatic cancer. Lancet 378, 607–620 (2011).
Agbunag, C., Lee, K.E., Buontempo, S. & Bar-Sagi, D. Pancreatic duct epithelial cell isolation and cultivation in two-dimensional and three-dimensional culture systems. Methods Enzymol. 407, 703–710 (2006).
Huch, M. et al. Unlimited in vitro expansion of adult bi-potent pancreas progenitors through the Lgr5/R-spondin axis. EMBO J. 32, 2708–2721 (2013).
Li, X. et al. Oncogenic transformation of diverse gastrointestinal tissues in primary organoid culture. Nat. Med. 20, 769–777 (2014).
Pagliuca, F.W. et al. Generation of functional human pancreatic beta cells in vitro. Cell 159, 428–439 (2014).
Rezania, A. et al. Reversal of diabetes with insulin-producing cells derived in vitro from human pluripotent stem cells. Nat. Biotechnol. 32, 1121–1133 (2014).
Schiesser, J.V. & Wells, J.M. Generation of beta cells from human pluripotent stem cells: are we there yet? Ann. NY Acad. Sci. 1311, 124–137 (2014).
Jennings, R.E. et al. Development of the human pancreas from foregut to endocrine commitment. Diabetes 62, 3514–3522 (2013).
Pan, F.C. & Wright, C. Pancreas organogenesis: from bud to plexus to gland. Dev. Dyn. 240, 530–565 (2011).
McCracken, K.W. & Wells, J.M. Molecular pathways controlling pancreas induction. Semin. Cell Dev. Biol. 23, 656–662 (2012).
Hick, A.C. et al. Mechanism of primitive duct formation in the pancreas and submandibular glands: a role for SDF-1. BMC Dev. Biol. 9, 66 (2009).
Riedel, M.J. et al. Immunohistochemical characterisation of cells co-producing insulin and glucagon in the developing human pancreas. Diabetologia 55, 372–381 (2012).
Lyttle, B.M. et al. Transcription factor expression in the developing human fetal endocrine pancreas. Diabetologia 51, 1169–1180 (2008).
Outzen, H.C. & Leiter, E.H. Transplantation of pancreatic islets into cleared mammary fat pads. Transplantation 32, 101–105 (1981).
Laitio, M., Lev, R. & Orlic, D. The developing human fetal pancreas: an ultrastructural and histochemical study with special reference to exocrine cells. J. Anat. 117, 619–634 (1974).
Nielsen, S.K. et al. Characterization of primary cilia and Hedgehog signaling during development of the human pancreas and in human pancreatic duct cancer cell lines. Dev. Dyn. 237, 2039–2052 (2008).
Kolodecik, T., Shugrue, C., Ashat, M. & Thrower, E.C. Risk factors for pancreatic cancer: underlying mechanisms and potential targets. Front. Physiol. 4, 415 (2013).
Chang, D.K., Grimmond, S.M. & Biankin, A.V. Pancreatic cancer genomics. Curr. Opin. Genet. Dev. 24, 74–81 (2014).
Chakravarty, G., Rider, B. & Mondal, D. Cytoplasmic compartmentalization of SOX9 abrogates the growth arrest response of breast cancer cells that can be rescued by trichostatin A treatment. Cancer Biol. Ther. 11, 71–83 (2011).
Chakravarty, G. et al. Prognostic significance of cytoplasmic SOX9 in invasive ductal carcinoma and metastatic breast cancer. Exp. Biol. Med. (Maywood) 236, 145–155 (2011).
Marusyk, A., Almendro, V. & Polyak, K. Intra-tumour heterogeneity: a looking glass for cancer? Nat. Rev. Cancer 12, 323–334 (2012).
Waddell, N. et al. Whole genomes redefine the mutational landscape of pancreatic cancer. Nature 518, 495–501 (2015).
van den Beucken, T. et al. Hypoxia promotes stem cell phenotypes and poor prognosis through epigenetic regulation of DICER. Nat. Commun. 5, 5203 (2014).
Johnson, A.B., Denko, N. & Barton, M.C. Hypoxia induces a novel signature of chromatin modifications and global repression of transcription. Mutat. Res. 640, 174–179 (2008).
Viale, A. et al. Oncogene ablation-resistant pancreatic cancer cells depend on mitochondrial function. Nature 514, 628–632 (2014).
Boj, S.F. et al. Organoid models of human and mouse ductal pancreatic cancer. Cell 160, 324–338 (2015).
Nostro, M.C. et al. Stage-specific signaling through TGF-β family members and WNT regulates patterning and pancreatic specification of human pluripotent stem cells. Development 138, 861–871 (2011).
Nostro, M.C. et al. Efficient generation of NKX6-1+ pancreatic progenitors from multiple human pluripotent stem cell lines. Stem Cell Reports 4, 591–604 (2015).
Xiang, B. & Muthuswamy, S.K. Using three-dimensional acinar structures for molecular and cell biological assays. Methods Enzymol. 406, 692–701 (2006).
Ritchie, M.E. et al. Limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 43, e47 (2015).
We thank members of S.K.M.'s laboratory for helpful suggestions and discussions, D. Barsyte-Lovejoy for helping with epigenetic drug screening experiments and members of the PanCuRx team, including D. Hedley, for support and assistance. This work was supported by the Ontario Institute for Cancer Research (OICR) PanCuRx program; Canadian Cancer Society; Lee K Margaret Lau Chair for Breast Cancer Research and Campbell Family Institute for Breast cancer research to S.K.M. The Structural Genomics Consortium is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada, Innovative Medicines Initiative (EU/EFPIA) (ULTRA-DD grant 115766), Janssen, Merck and Co., Novartis Pharma AG, Ontario Ministry of Economic Development and Innovation, Pfizer, São Paulo Research Foundation (FAPESP), Takeda, and the Wellcome Trust. This was also funded in part by the Ontario Ministry of Health and Long Term Care (OMOHLTC). The views expressed do not necessarily reflect those of the OMOHLTC.
The authors declare no competing financial interests.
Supplementary Figures 1–5 and Supplementary Tables 1–3 (PDF 1510 kb)
Supplementary source data (XLSX 11 kb)
About this article
Cite this article
Huang, L., Holtzinger, A., Jagan, I. et al. Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell– and patient-derived tumor organoids. Nat Med 21, 1364–1371 (2015). https://doi.org/10.1038/nm.3973
Developmental Dynamics (2020)
Nature Reviews Gastroenterology & Hepatology (2020)
Generation and characterization of a cell line from an intraductal tubulopapillary neoplasm of the pancreas
Laboratory Investigation (2020)
Bioinformatics and computational approaches for analyzing patient-derived disease models in cancer research
Computational and Structural Biotechnology Journal (2020)