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

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Organoids
Barrett Esophagus
Neoplasms
Catenins
Biopsy
Goblet Cells
Gene Editing
Basement Membrane
Noise
Cell Differentiation
Adenocarcinoma
Genome
Apoptosis
Viruses
Cell Line

Keywords

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

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Modeling Wnt signaling by CRISPR-Cas9 genome editing recapitulates neoplasia in human Barrett epithelial organoids. / Liu, Xi; Cheng, Yulan; Abraham, John M.; Wang, Zhixiong; Wang, Zhe; Ke, Xiquan; Yan, Rong; Shin, Eun; Ngamruengphong, Saowanee; Khashab, Mouen; Zhang, Guanjun; McNamara, George; Ewald, Andrew; Lin, De Chen; Liu, Zhengwen; Meltzer, Stephen.

In: Cancer Letters, Vol. 436, 01.11.2018, p. 109-118.

Research output: Contribution to journalArticle

Liu, Xi ; Cheng, Yulan ; Abraham, John M. ; Wang, Zhixiong ; Wang, Zhe ; Ke, Xiquan ; Yan, Rong ; Shin, Eun ; Ngamruengphong, Saowanee ; Khashab, Mouen ; Zhang, Guanjun ; McNamara, George ; Ewald, Andrew ; Lin, De Chen ; Liu, Zhengwen ; Meltzer, Stephen. / Modeling Wnt signaling by CRISPR-Cas9 genome editing recapitulates neoplasia in human Barrett epithelial organoids. In: Cancer Letters. 2018 ; Vol. 436. pp. 109-118.
@article{8ad4ed05bb064355a9ba4e4daee0aaf7,
title = "Modeling Wnt signaling by CRISPR-Cas9 genome editing recapitulates neoplasia in human Barrett epithelial organoids",
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.",
keywords = "Barrett esophagus, CRISPR/Cas9, Neoplastic transformation, Organoids, Wnt signaling",
author = "Xi Liu and Yulan Cheng and Abraham, {John M.} and Zhixiong Wang and Zhe Wang and Xiquan Ke and Rong Yan and Eun Shin and Saowanee Ngamruengphong and Mouen Khashab and Guanjun Zhang and George McNamara and Andrew Ewald and Lin, {De Chen} and Zhengwen Liu and Stephen Meltzer",
year = "2018",
month = "11",
day = "1",
doi = "10.1016/j.canlet.2018.08.017",
language = "English (US)",
volume = "436",
pages = "109--118",
journal = "Cancer Letters",
issn = "0304-3835",
publisher = "Elsevier Ireland Ltd",

}

TY - JOUR

T1 - Modeling Wnt signaling by CRISPR-Cas9 genome editing recapitulates neoplasia in human Barrett epithelial organoids

AU - Liu, Xi

AU - Cheng, Yulan

AU - Abraham, John M.

AU - Wang, Zhixiong

AU - Wang, Zhe

AU - Ke, Xiquan

AU - Yan, Rong

AU - Shin, Eun

AU - Ngamruengphong, Saowanee

AU - Khashab, Mouen

AU - Zhang, Guanjun

AU - McNamara, George

AU - Ewald, Andrew

AU - Lin, De Chen

AU - Liu, Zhengwen

AU - Meltzer, Stephen

PY - 2018/11/1

Y1 - 2018/11/1

N2 - 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.

AB - 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.

KW - Barrett esophagus

KW - CRISPR/Cas9

KW - Neoplastic transformation

KW - Organoids

KW - Wnt signaling

UR - http://www.scopus.com/inward/record.url?scp=85052316463&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85052316463&partnerID=8YFLogxK

U2 - 10.1016/j.canlet.2018.08.017

DO - 10.1016/j.canlet.2018.08.017

M3 - Article

C2 - 30144514

AN - SCOPUS:85052316463

VL - 436

SP - 109

EP - 118

JO - Cancer Letters

JF - Cancer Letters

SN - 0304-3835

ER -