Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell- and patient-derived tumor organoids.
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.
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.
Ling
L
Huang
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.|McEwen Center for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada.
Audrey
A
Holtzinger
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.
Ishaan
I
Jagan
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.
Michael
M
BeGora
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.
Ines
I
Lohse
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.
Nicholas
N
Ngai
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.|McEwen Center for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada.
Cristina
C
Nostro
Department of Physiology, Western University, London, Ontario, Canada.|Department of Pharmacology, Western University, London, Ontario, Canada.
Rennian
R
Wang
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.
Lakshmi B
LB
Muthuswamy
Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, Michigan.|Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan.
Howard C
HC
Crawford
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.|Structural Genomics Consortium, Toronto, Ontario, Canada.
Cheryl
C
Arrowsmith
Division of Anatomic Pathology, Vancouver General Hospital, Vancouver, British Columbia, Canada.|Department of Medicine, The University of British Columbia, Vancouver, British Columbia, Canada.|Pancreas Centre British Columbia, Vancouver, British Columbia, Canada.
Steve E
SE
Kalloger
Department of Medicine, The University of British Columbia, Vancouver, British Columbia, Canada.|Pancreas Centre British Columbia, Vancouver, British Columbia, Canada.|Division of Medical Oncology, British Columbia Cancer Agency, Vancouver, British Columbia, Canada.
Daniel J
DJ
Renouf
Division of General Surgery, University of Toronto, Toronto, Ontario, Canada.
Ashton A
AA
Connor
Division of General Surgery, University of Toronto, Toronto, Ontario, Canada.
Sean
S
Cleary
Division of Anatomic Pathology, Vancouver General Hospital, Vancouver, British Columbia, Canada.|Department of Medicine, The University of British Columbia, Vancouver, British Columbia, Canada.|Pancreas Centre British Columbia, Vancouver, British Columbia, Canada.
David F
DF
Schaeffer
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.
Michael
M
Roehrl
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.|Department of Pathology, University Health Network, Toronto, Ontario, Canada.
Ming-Sound
MS
Tsao
Division of General Surgery, University of Toronto, Toronto, Ontario, Canada.
Steven
S
Gallinger
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.|McEwen Center for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada.
Gordon
G
Keller
Princess Margaret Cancer Center, University Health Network (UHN), University of Toronto, Toronto, Ontario, Canada.
Senthil K
SK
Muthuswamy
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D021441:Carcinoma, Pancreatic Ductal / Q000188:drug therapy* / Q000235:genetics; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D006801:Humans; D051379:Mice; D008954:Models, Biological; D009154:Mutation; D009940:Organoids / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure; D010179:Pancreas / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000235:genetics; D039904:Pluripotent Stem Cells; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras); D055758:SOX9 Transcription Factor / Q000378:metabolism; D046509:Tissue Culture Techniques; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D018631:ras Proteins / Q000235:genetics; D000093542:Gemcitabine
D000964:Antimetabolites, Antineoplastic; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D003841:Deoxycytidine; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins; D000093542:Gemcitabine