Cargando…

APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas

BACKGROUND: Esophageal adenocarcinoma (EAC) is characterized by poor prognosis and low survival rate. Chronic gastroesophageal reflux disease (GERD) is the main risk factor for the development of Barrett’s esophagus (BE), a preneoplastic metaplastic condition, and its progression to EAC. Yes-associa...

Descripción completa

Detalles Bibliográficos
Autores principales: Ballout, Farah, Lu, Heng, Chen, Lei, Sriramajayam, Kannappan, Que, Jianwen, Meng, Zhipeng, Wang, Timothy C., Giordano, Silvia, Zaika, Alexander, McDonald, Oliver, Peng, Dunfa, El-Rifai, Wael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9434868/
https://www.ncbi.nlm.nih.gov/pubmed/36045416
http://dx.doi.org/10.1186/s13046-022-02472-5
_version_ 1784780978180325376
author Ballout, Farah
Lu, Heng
Chen, Lei
Sriramajayam, Kannappan
Que, Jianwen
Meng, Zhipeng
Wang, Timothy C.
Giordano, Silvia
Zaika, Alexander
McDonald, Oliver
Peng, Dunfa
El-Rifai, Wael
author_facet Ballout, Farah
Lu, Heng
Chen, Lei
Sriramajayam, Kannappan
Que, Jianwen
Meng, Zhipeng
Wang, Timothy C.
Giordano, Silvia
Zaika, Alexander
McDonald, Oliver
Peng, Dunfa
El-Rifai, Wael
author_sort Ballout, Farah
collection PubMed
description BACKGROUND: Esophageal adenocarcinoma (EAC) is characterized by poor prognosis and low survival rate. Chronic gastroesophageal reflux disease (GERD) is the main risk factor for the development of Barrett’s esophagus (BE), a preneoplastic metaplastic condition, and its progression to EAC. Yes-associated protein 1 (YAP1) activation mediates stem-like properties under cellular stress. The role of acidic bile salts (ABS) in promoting YAP1 activation under reflux conditions remains unexplored. METHODS: A combination of EAC cell lines, transgenic mice, and patient-derived xenografts were utilized in this study. mRNA expression and protein levels of APE1 and YAP1 were evaluated by qRT-PCR, western blot, and immunohistochemistry. YAP1 activation was confirmed by immunofluorescence staining and luciferase transcriptional activity reporter assay. The functional role and mechanism of regulation of YAP1 by APE1 was determined by sphere formation assay, siRNA mediated knockdown, redox-specific inhibition, and co-immunoprecipitation assays. RESULTS: We showed that YAP1 signaling is activated in BE and EAC cells following exposure to ABS, the mimicry of reflux conditions in patients with GERD. This induction was consistent with APE1 upregulation in response to ABS. YAP1 activation was confirmed by its nuclear accumulation with corresponding up-regulation of YAP1 target genes. APE1 silencing inhibited YAP1 protein induction and reduced its nuclear expression and transcriptional activity, following ABS treatment. Further investigation revealed that APE1-redox-specific inhibition (E3330) or APE1 redox-deficient mutant (C65A) abrogated ABS-mediated YAP1 activation, indicating an APE1 redox-dependent mechanism. APE1 silencing or E3330 treatment reduced YAP1 protein levels and diminished the number and size of EAC spheroids. Mechanistically, we demonstrated that APE1 regulated YAP1 stability through interaction with β-TrCP ubiquitinase, whereas APE1-redox-specific inhibition induced YAP1 poly-ubiquitination promoting its degradation. CONCLUSION: Our findings established a novel function of APE1 in EAC progression elucidating druggable molecular vulnerabilities via targeting APE1 or YAP1 for the treatment of EAC. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-022-02472-5.
format Online
Article
Text
id pubmed-9434868
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-94348682022-09-02 APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas Ballout, Farah Lu, Heng Chen, Lei Sriramajayam, Kannappan Que, Jianwen Meng, Zhipeng Wang, Timothy C. Giordano, Silvia Zaika, Alexander McDonald, Oliver Peng, Dunfa El-Rifai, Wael J Exp Clin Cancer Res Research BACKGROUND: Esophageal adenocarcinoma (EAC) is characterized by poor prognosis and low survival rate. Chronic gastroesophageal reflux disease (GERD) is the main risk factor for the development of Barrett’s esophagus (BE), a preneoplastic metaplastic condition, and its progression to EAC. Yes-associated protein 1 (YAP1) activation mediates stem-like properties under cellular stress. The role of acidic bile salts (ABS) in promoting YAP1 activation under reflux conditions remains unexplored. METHODS: A combination of EAC cell lines, transgenic mice, and patient-derived xenografts were utilized in this study. mRNA expression and protein levels of APE1 and YAP1 were evaluated by qRT-PCR, western blot, and immunohistochemistry. YAP1 activation was confirmed by immunofluorescence staining and luciferase transcriptional activity reporter assay. The functional role and mechanism of regulation of YAP1 by APE1 was determined by sphere formation assay, siRNA mediated knockdown, redox-specific inhibition, and co-immunoprecipitation assays. RESULTS: We showed that YAP1 signaling is activated in BE and EAC cells following exposure to ABS, the mimicry of reflux conditions in patients with GERD. This induction was consistent with APE1 upregulation in response to ABS. YAP1 activation was confirmed by its nuclear accumulation with corresponding up-regulation of YAP1 target genes. APE1 silencing inhibited YAP1 protein induction and reduced its nuclear expression and transcriptional activity, following ABS treatment. Further investigation revealed that APE1-redox-specific inhibition (E3330) or APE1 redox-deficient mutant (C65A) abrogated ABS-mediated YAP1 activation, indicating an APE1 redox-dependent mechanism. APE1 silencing or E3330 treatment reduced YAP1 protein levels and diminished the number and size of EAC spheroids. Mechanistically, we demonstrated that APE1 regulated YAP1 stability through interaction with β-TrCP ubiquitinase, whereas APE1-redox-specific inhibition induced YAP1 poly-ubiquitination promoting its degradation. CONCLUSION: Our findings established a novel function of APE1 in EAC progression elucidating druggable molecular vulnerabilities via targeting APE1 or YAP1 for the treatment of EAC. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13046-022-02472-5. BioMed Central 2022-09-01 /pmc/articles/PMC9434868/ /pubmed/36045416 http://dx.doi.org/10.1186/s13046-022-02472-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Ballout, Farah
Lu, Heng
Chen, Lei
Sriramajayam, Kannappan
Que, Jianwen
Meng, Zhipeng
Wang, Timothy C.
Giordano, Silvia
Zaika, Alexander
McDonald, Oliver
Peng, Dunfa
El-Rifai, Wael
APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas
title APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas
title_full APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas
title_fullStr APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas
title_full_unstemmed APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas
title_short APE1 redox function is required for activation of Yes-associated protein 1 under reflux conditions in Barrett’s-associated esophageal adenocarcinomas
title_sort ape1 redox function is required for activation of yes-associated protein 1 under reflux conditions in barrett’s-associated esophageal adenocarcinomas
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9434868/
https://www.ncbi.nlm.nih.gov/pubmed/36045416
http://dx.doi.org/10.1186/s13046-022-02472-5
work_keys_str_mv AT balloutfarah ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT luheng ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT chenlei ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT sriramajayamkannappan ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT quejianwen ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT mengzhipeng ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT wangtimothyc ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT giordanosilvia ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT zaikaalexander ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT mcdonaldoliver ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT pengdunfa ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas
AT elrifaiwael ape1redoxfunctionisrequiredforactivationofyesassociatedprotein1underrefluxconditionsinbarrettsassociatedesophagealadenocarcinomas