Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett’s EsophagusSummary

Background & Aims: Esophageal adenocarcinoma (EAC) develops from its precursor Barrett’s esophagus through intermediate stages of low- and high-grade dysplasia. However, knowledge of genetic drivers and molecular mechanisms implicated in disease progression is limited. Herein, we investigate...

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Main Authors: Jovana R. Gotovac, Tanjina Kader, Julia V. Milne, Kenji M. Fujihara, Luis E. Lara-Gonzalez, Kylie L. Gorringe, Sangeetha N. Kalimuthu, Madawa W. Jayawardana, Cuong P. Duong, Wayne A. Phillips, Nicholas J. Clemons
Format: Article
Language:English
Published: Elsevier 2021-01-01
Series:Cellular and Molecular Gastroenterology and Hepatology
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2352345X21000606
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author Jovana R. Gotovac
Tanjina Kader
Julia V. Milne
Kenji M. Fujihara
Luis E. Lara-Gonzalez
Kylie L. Gorringe
Sangeetha N. Kalimuthu
Madawa W. Jayawardana
Cuong P. Duong
Wayne A. Phillips
Nicholas J. Clemons
author_facet Jovana R. Gotovac
Tanjina Kader
Julia V. Milne
Kenji M. Fujihara
Luis E. Lara-Gonzalez
Kylie L. Gorringe
Sangeetha N. Kalimuthu
Madawa W. Jayawardana
Cuong P. Duong
Wayne A. Phillips
Nicholas J. Clemons
author_sort Jovana R. Gotovac
collection DOAJ
description Background & Aims: Esophageal adenocarcinoma (EAC) develops from its precursor Barrett’s esophagus through intermediate stages of low- and high-grade dysplasia. However, knowledge of genetic drivers and molecular mechanisms implicated in disease progression is limited. Herein, we investigated the effect of Mothers against decapentaplegic homolog 4 (SMAD4) loss on transforming growth factor β (TGF-β) signaling functionality and in vivo tumorigenicity in high-grade dysplastic Barrett’s cells. Methods: An in vivo xenograft model was used to test tumorigenicity of SMAD4 knockdown or knockout in CP-B high-grade dysplastic Barrett’s cells. RT2 polymerase chain reaction arrays were used to analyze TGF-β signaling functionality, and low-coverage whole-genome sequencing was performed to detect copy number alterations upon SMAD4 loss. Results: We found that SMAD4 knockout significantly alters the TGF-β pathway target gene expression profile. SMAD4 knockout positively regulates potential oncogenes such as CRYAB, ACTA2, and CDC6, whereas the CDKN2A/B tumor-suppressor locus was regulated negatively. We verified that SMAD4 in combination with CDC6-CDKN2A/B or CRYAB genetic alterations in patient tumors have significant predictive value for poor prognosis. Importantly, we investigated the effect of SMAD4 inactivation in Barrett’s tumorigenesis. We found that genetic knockdown or knockout of SMAD4 was sufficient to promote tumorigenesis in dysplastic Barrett’s esophagus cells in vivo. Progression to invasive EAC was accompanied by distinctive and consistent copy number alterations in SMAD4 knockdown or knockout xenografts. Conclusions: Altogether, up-regulation of oncogenes, down-regulation of tumor-suppressor genes, and chromosomal instability within the tumors after SMAD4 loss implicates SMAD4 as a protector of genome integrity in EAC development and progression. Foremost, SMAD4 loss promotes tumorigenesis from dysplastic Barrett’s toward EAC.
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spelling doaj.art-a7e5902c1bfa452b8729ab028422f9392022-12-21T18:22:14ZengElsevierCellular and Molecular Gastroenterology and Hepatology2352-345X2021-01-01122689713Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett’s EsophagusSummaryJovana R. Gotovac0Tanjina Kader1Julia V. Milne2Kenji M. Fujihara3Luis E. Lara-Gonzalez4Kylie L. Gorringe5Sangeetha N. Kalimuthu6Madawa W. Jayawardana7Cuong P. Duong8Wayne A. Phillips9Nicholas J. Clemons10Division of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, AustraliaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, AustraliaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, AustraliaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, AustraliaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, AustraliaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, AustraliaAnatomical Pathology, Laboratory Medicine Program, University Health Network, Toronto, Ontario, Canada; Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, CanadaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, AustraliaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, AustraliaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, Australia; Department of Surgery, St Vincent’s Hospital, The University of Melbourne, Parkville, Victoria, AustraliaDivision of Cancer Research, Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, Parkville, Victoria, Australia; Correspondence Address correspondence to: Nicholas J. Clemons, PhD, Division of Cancer Research, Peter MacCallum Cancer Centre, 305 Grattan Street, Melbourne, Victoria 3000, Australia. fax: (61) 3-8559-7379.Background & Aims: Esophageal adenocarcinoma (EAC) develops from its precursor Barrett’s esophagus through intermediate stages of low- and high-grade dysplasia. However, knowledge of genetic drivers and molecular mechanisms implicated in disease progression is limited. Herein, we investigated the effect of Mothers against decapentaplegic homolog 4 (SMAD4) loss on transforming growth factor β (TGF-β) signaling functionality and in vivo tumorigenicity in high-grade dysplastic Barrett’s cells. Methods: An in vivo xenograft model was used to test tumorigenicity of SMAD4 knockdown or knockout in CP-B high-grade dysplastic Barrett’s cells. RT2 polymerase chain reaction arrays were used to analyze TGF-β signaling functionality, and low-coverage whole-genome sequencing was performed to detect copy number alterations upon SMAD4 loss. Results: We found that SMAD4 knockout significantly alters the TGF-β pathway target gene expression profile. SMAD4 knockout positively regulates potential oncogenes such as CRYAB, ACTA2, and CDC6, whereas the CDKN2A/B tumor-suppressor locus was regulated negatively. We verified that SMAD4 in combination with CDC6-CDKN2A/B or CRYAB genetic alterations in patient tumors have significant predictive value for poor prognosis. Importantly, we investigated the effect of SMAD4 inactivation in Barrett’s tumorigenesis. We found that genetic knockdown or knockout of SMAD4 was sufficient to promote tumorigenesis in dysplastic Barrett’s esophagus cells in vivo. Progression to invasive EAC was accompanied by distinctive and consistent copy number alterations in SMAD4 knockdown or knockout xenografts. Conclusions: Altogether, up-regulation of oncogenes, down-regulation of tumor-suppressor genes, and chromosomal instability within the tumors after SMAD4 loss implicates SMAD4 as a protector of genome integrity in EAC development and progression. Foremost, SMAD4 loss promotes tumorigenesis from dysplastic Barrett’s toward EAC.http://www.sciencedirect.com/science/article/pii/S2352345X21000606Esophageal Adenocarcinoma (EAC)SMAD4 LossCopy Number Alterations (CNA)Barrett’s Tumorigenesis
spellingShingle Jovana R. Gotovac
Tanjina Kader
Julia V. Milne
Kenji M. Fujihara
Luis E. Lara-Gonzalez
Kylie L. Gorringe
Sangeetha N. Kalimuthu
Madawa W. Jayawardana
Cuong P. Duong
Wayne A. Phillips
Nicholas J. Clemons
Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett’s EsophagusSummary
Cellular and Molecular Gastroenterology and Hepatology
Esophageal Adenocarcinoma (EAC)
SMAD4 Loss
Copy Number Alterations (CNA)
Barrett’s Tumorigenesis
title Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett’s EsophagusSummary
title_full Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett’s EsophagusSummary
title_fullStr Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett’s EsophagusSummary
title_full_unstemmed Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett’s EsophagusSummary
title_short Loss of SMAD4 Is Sufficient to Promote Tumorigenesis in a Model of Dysplastic Barrett’s EsophagusSummary
title_sort loss of smad4 is sufficient to promote tumorigenesis in a model of dysplastic barrett s esophagussummary
topic Esophageal Adenocarcinoma (EAC)
SMAD4 Loss
Copy Number Alterations (CNA)
Barrett’s Tumorigenesis
url http://www.sciencedirect.com/science/article/pii/S2352345X21000606
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