title: Oncogenic transformation of diverse gastrointestinal tissues in primary organoid culture.
abstract: The application of primary organoid cultures containing epithelial and mesenchymal elements to cancer modeling holds promise for combining the accurate multilineage differentiation and physiology of in vivo systems with the facile in vitro manipulation of transformed cell lines. Here we used a single air-liquid interface culture method without modification to engineer oncogenic mutations into primary epithelial and mesenchymal organoids from mouse colon, stomach and pancreas. Pancreatic and gastric organoids exhibited dysplasia as a result of expression of Kras carrying the G12D mutation (Kras(G12D)), p53 loss or both and readily generated adenocarcinoma after in vivo transplantation. In contrast, primary colon organoids required combinatorial Apc, p53, Kras(G12D) and Smad4 mutations for progressive transformation to invasive adenocarcinoma-like histology in vitro and tumorigenicity in vivo, recapitulating multi-hit models of colorectal cancer (CRC), as compared to the more promiscuous transformation of small intestinal organoids. Colon organoid culture functionally validated the microRNA miR-483 as a dominant driver oncogene at the IGF2 (insulin-like growth factor-2) 11p15.5 CRC amplicon, inducing dysplasia in vitro and tumorigenicity in vivo. These studies demonstrate the general utility of a highly tractable primary organoid system for cancer modeling and driver oncogene validation in diverse gastrointestinal tissues.
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Xingnan
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Lincoln
authors.initials: L
authors.lastname: Nadauld
authors.affiliation: 1] Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA. [2] Department of Internal Medicine, Saga Medical School, Saga, Japan.
authors.forename: Akifumi
authors.initials: A
authors.lastname: Ootani
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: David C
authors.initials: DC
authors.lastname: Corney
authors.affiliation: Department of Pathology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Reetesh K
authors.initials: RK
authors.lastname: Pai
authors.affiliation: Department of Radiology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Olivier
authors.initials: O
authors.lastname: Gevaert
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Cantrell
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Paul G
authors.initials: PG
authors.lastname: Rack
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: James T
authors.initials: JT
authors.lastname: Neal
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Carol W-M
authors.initials: CW
authors.lastname: Chan
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Trevor
authors.initials: T
authors.lastname: Yeung
authors.affiliation: Baxter Laboratories, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Xue
authors.initials: X
authors.lastname: Gong
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Jenny
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Division of Oncology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Wilhelmy
authors.affiliation: Equipe de Morphogenèse et Signalisation Cellulaires, UMR 144 CNRS/Institut Curie, Paris, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Robine
authors.affiliation: Division of Radiation Biology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Laura D
authors.initials: LD
authors.lastname: Attardi
authors.affiliation: Department of Radiology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Sylvia K
authors.initials: SK
authors.lastname: Plevritis
authors.affiliation: Department of Medicine, Sackler School of Graduate Biomedical Sciences, Tufts University School of Medicine, Boston, Massachusetts, USA.
authors.forename: Kenneth E
authors.initials: KE
authors.lastname: Hung
authors.affiliation: Baxter Laboratories, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Chang-Zheng
authors.initials: CZ
authors.lastname: Chen
authors.affiliation: Division of Oncology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Hanlee P
authors.initials: HP
authors.lastname: Ji
authors.affiliation: Department of Medicine, Hematology Division, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Calvin J
authors.initials: CJ
authors.lastname: Kuo
mesh_terms: D000818:Animals; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D005770:Gastrointestinal Neoplasms / Q000473:pathology; D041981:Gastrointestinal Tract / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009857:Oncogenes; D009924:Organ Culture Techniques