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

Nat Med. 2015 Nov;21(11):1364-71. doi: 10.1038/nm.3973. Epub 2015 Oct 26.

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 TP53(R175H) 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antimetabolites, Antineoplastic / pharmacology*
  • Carcinoma, Pancreatic Ductal / drug therapy*
  • Carcinoma, Pancreatic Ductal / genetics
  • Deoxycytidine / analogs & derivatives*
  • Deoxycytidine / pharmacology
  • Drug Screening Assays, Antitumor / methods
  • Humans
  • Mice
  • Models, Biological
  • Mutation
  • Organoids / drug effects*
  • Organoids / pathology
  • Organoids / ultrastructure
  • Pancreas / drug effects*
  • Pancreas / pathology
  • Pancreas / ultrastructure
  • Pancreatic Neoplasms / drug therapy*
  • Pancreatic Neoplasms / genetics
  • Pluripotent Stem Cells*
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins p21(ras)
  • SOX9 Transcription Factor / metabolism
  • Tissue Culture Techniques
  • Tumor Suppressor Protein p53 / genetics
  • ras Proteins / genetics

Substances

  • Antimetabolites, Antineoplastic
  • KRAS protein, human
  • Proto-Oncogene Proteins
  • SOX9 Transcription Factor
  • SOX9 protein, human
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • Deoxycytidine
  • gemcitabine
  • Proto-Oncogene Proteins p21(ras)
  • ras Proteins