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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Elsevier
2021-01-01
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Series: | Cellular and Molecular Gastroenterology and Hepatology |
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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. |
first_indexed | 2024-12-22T14:54:59Z |
format | Article |
id | doaj.art-a7e5902c1bfa452b8729ab028422f939 |
institution | Directory Open Access Journal |
issn | 2352-345X |
language | English |
last_indexed | 2024-12-22T14:54:59Z |
publishDate | 2021-01-01 |
publisher | Elsevier |
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series | Cellular and Molecular Gastroenterology and Hepatology |
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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