Modeling Wnt signaling by CRISPR-Cas9 genome editing recapitulates neoplasia in human Barrett epithelial organoids

Xi Liu, Yulan Cheng, John M. Abraham, Zhixiong Wang, Zhe Wang, Xiquan Ke, Rong Yan, Eun Shin, Saowanee Ngamruengphong, Mouen Khashab, Guanjun Zhang, George McNamara, Andrew Ewald, De Chen Lin, Zhengwen Liu, Stephen Meltzer

Research output: Contribution to journalArticle

Abstract

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 (APCKO) BE organoids genome-edited by CRISPR-Cas9 showed characteristic goblet cell differentiation. Autonomous Wnt activation was confirmed in APCKO organoids by overexpression of Wnt target genes and nuclear-translocated β-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/β-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.

Original languageEnglish (US)
Pages (from-to)109-118
Number of pages10
JournalCancer Letters
Volume436
DOIs
StatePublished - Nov 1 2018

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Keywords

  • Barrett esophagus
  • CRISPR/Cas9
  • Neoplastic transformation
  • Organoids
  • Wnt signaling

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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