Journal
CANCER LETTERS
Volume 436, Issue -, Pages 109-118Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.canlet.2018.08.017
Keywords
Organoids; Barrett esophagus; Wnt signaling; CRISPR/Cas9; Neoplastic transformation
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Funding
- National Institutes of Health [CA190040, CA211457]
- Johns Hopkins University Discovery Fund
- Key Science and Technology Program of Shaanxi Province, China [2015SF128]
- International Scientific and Technological Cooperation and Exchange Program of Shaanxi Province, China [2015KW-030]
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Primary organoid cultures generated from patient biopsies comprise a novel improved platform for disease modeling, being genetically stable and closely recapitulating in vivo scenarios. Barrett esophagus (BE) is the major risk factor for esophageal adenocarcinoma. There has been a dearth of long-term in vitro expansion models of BE neoplastic transformation. We generated a long-term virus-free organoid expansion model of BE neoplasia from patient biopsies. Both wild-type and paired APC-knockout (APC(KO)) BE organoids genome-edited by CRISPR-Cas9 showed characteristic goblet cell differentiation. Autonomous Wnt activation was confirmed in APC(KO) organoids by overexpression of Wnt target genes and nuclear-translocated beta-catenin expression after withdrawal of Wnt-3A and R-spondin-1. Wnt-activated organoids demonstrated histologic atypia, higher proliferative and replicative activity, reduced apoptosis, and prolonged culturability. Wnt-activated organoids also showed sustained protrusive migration ability accompanied by disrupted basement membrane reorganization and integrity. This CRISPR-Cas9 editing human-derived organoid model recapitulates the critical role of aberrant Wnt/beta-catenin signaling activation in BE neoplastic transformation. This system can be used to study other 'driver' pathway alterations in BE-associated neoplasia, avoiding signaling noise present in immortalized or cancer-derived cell lines.
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