Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell– and patient-derived tumor organoids

Abstract

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.

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Figure 1: Induction of polarized organoids from human pluripotent stem cells.
Figure 2: Organoids express markers associated with pancreatic progenitor cells.
Figure 3: Differentiation of pancreatic progenitor organoids in vitro and in vivo.
Figure 4: Progenitor organoids model early PDAC.
Figure 5: TP53 mutational status, localization of SOX9 and clinical outcome.
Figure 6: Establishment of tumor organoids that conserve patient-specific traits.

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Acknowledgements

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.

Author information

S.K.M. conceived and coordinated the project, designed experiments and co-wrote the manuscript with L.H. L.H. also designed, performed and/or coordinated all experiments. G.K. and A.H. contributed to pancreatic lineage–committed precursor generation. I.J. contributed tumor organoid live imaging and manuscript preparation. M.B. contributed to tumor organoid immunofluorescence microscopy. I.L. contributed to collection of patient-derived xenograft tumors. N.N. contributed to organoid size measurement. C.N. contributed to pancreatic lineage–committed precursor generation. R.W. contributed to human fetal pancreas studies. L.B.M. contributed to bioinformatics analysis for gene expressions. H.C.C. contributed to experimental design. C.A. contributed to epigenetic drug screening. S.E.K., D.J.R., A.A.C., S.C. and D.F.S. contributed to studies of P53 and SOX9 localization in patient samples. M.R. contributed to pathological analysis on patient tumor and tumor organoids, and studies of P53 and SOX9 localization in patient samples. M.-S.T. contributed to pathological analysis on patient tumor and tumor organoids, and studies of P53 and SOX9 localization in patient samples. S.G. contributed to obtaining patient resections for tumor organoid.

Correspondence to Senthil K Muthuswamy.

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The authors declare no competing financial interests.

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Time-lapse imaging of UHN6 tumor-organoid (AVI 17147 kb)

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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

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