Colonic organoids derived from human induced pluripotent stem cells for modeling colorectal cancer and drug testing
Journal
Nature Medicine
Journal Volume
23
Journal Issue
7
Pages
878 +
Date Issued
2017
Author(s)
M. Crespo
E. Vilar
K. Chang
S. Amin
T. Srinivasan
T. Zhang
N. H. Pipalia
H. J. Chen
M. Witherspoon
M. Gordillo
J. Z. Xiang
F. R. Maxfield
S. Lipkin
T. Evans
S. B. Chen
Abstract
With the goal of modeling human disease of the large intestine, we sought to develop an effective protocol for deriving colonic organoids (COs) from differentiated human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs). Extensive gene and immunohistochemical profiling confirmed that the derived COs represent colon rather than small intestine, containing stem cells, transit-amplifying cells, and the expected spectrum of differentiated cells, including goblet and endocrine cells. We applied this strategy to iPSCs derived from patients with familial adenomatous polyposis (FAP-iPSCs) harboring germline mutations in the WNT-signaling-pathway-regulator gene encoding APC, and we generated COs that exhibit enhanced WNT activity and increased epithelial cell proliferation, which we used as a platform for drug testing. Two potential compounds, XAV939 and rapamycin, decreased proliferation in FAP-COs, but also affected cell proliferation in wild-type COs, which thus limits their therapeutic application. By contrast, we found that geneticin, a ribosome-binding antibiotic with translational 'read-through' activity, efficiently targeted abnormal WNT activity and restored normal proliferation specifically in APC-mutant FAP-COs. These studies provide an efficient strategy for deriving human COs, which can be used in disease modeling and drug discovery for colorectal disease. ? 2017 Nature America, Inc., part of Springer Nature. All rights reserved.
SDGs
Other Subjects
7,8 dihydro 2 [4 (trifluoromethyl)phenyl] 5h thiopyrano[4,3 d]pyrimidin 4 ol; antibiotic g 418; APC protein; rapamycin; 7,8 dihydro 2 [4 (trifluoromethyl)phenyl] 5h thiopyrano[4,3 d]pyrimidin 4 ol; antineoplastic antibiotic; APC protein; APC protein, human; fused heterocyclic rings; gentamicin; rapamycin; antiproliferative activity; Article; cancer model; cell differentiation; cell proliferation; colorectal cancer; controlled study; cytoplasm; drug screening; embryo; epithelium cell; familial adenomatous polyposis; gene expression; germline mutation; human; human cell; human embryonic stem cell; induced pluripotent stem cell; priority journal; wild type; Wnt signaling pathway; adenoma; colon; colon polyposis; colorectal tumor; confocal microscopy; cytology; cytoplasm; drug effects; drug screening; enteroendocrine cell; flow cytometry; fluorescent antibody technique; gene expression profiling; genetics; goblet cell; human embryonic stem cell; immunohistochemistry; induced pluripotent stem cell; metabolism; mutation; pathology; real time polymerase chain reaction; Western blotting; Wnt signaling; Adenoma; Adenomatous Polyposis Coli; Adenomatous Polyposis Coli Protein; Antibiotics, Antineoplastic; Blotting, Western; Cell Differentiation; Cell Proliferation; Colon; Colorectal Neoplasms; Drug Screening Assays, Antitumor; Enteroendocrine Cells; Flow Cytometry; Fluorescent Antibody Technique; Gene Expression Profiling; Gentamicins; Germ-Line Mutation; Goblet Cells; Heterocyclic Compounds, 3-Ring; Human Embryonic Stem Cells; Humans; Immunohistochemistry; Induced Pluripotent Stem Cells; Microscopy, Confocal; Mutation; Organoids; Real-Time Polymerase Chain Reaction; Sirolimus; Wnt Signaling Pathway
Type
journal article