A novel ERβ high throughput microscopy platform for testing endocrine disrupting chemicals
In this study we present an inducible biosensor model for the Estrogen Receptor Beta (ERβ), GFP-ERβ:PRL-HeLa, a single-cell-based high throughput (HT) in vitro assay that allows direct visualization and measurement of GFP-tagged ERβ binding to ER-specific DNA response elements (EREs), ERβ-induced ch...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2024-01-01
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| Series: | Heliyon |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844023103276 |
| _version_ | 1797337097718202368 |
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| author | Derek A. Abbott Maureen G. Mancini Michael J. Bolt Adam T. Szafran Kaley A. Neugebauer Fabio Stossi Daniel A. Gorelick Michael A. Mancini |
| author_facet | Derek A. Abbott Maureen G. Mancini Michael J. Bolt Adam T. Szafran Kaley A. Neugebauer Fabio Stossi Daniel A. Gorelick Michael A. Mancini |
| author_sort | Derek A. Abbott |
| collection | DOAJ |
| description | In this study we present an inducible biosensor model for the Estrogen Receptor Beta (ERβ), GFP-ERβ:PRL-HeLa, a single-cell-based high throughput (HT) in vitro assay that allows direct visualization and measurement of GFP-tagged ERβ binding to ER-specific DNA response elements (EREs), ERβ-induced chromatin remodeling, and monitor transcriptional alterations via mRNA fluorescence in situ hybridization for a prolactin (PRL)-dsRED2 reporter gene. The model was used to accurately (Z’ = 0.58–0.8) differentiate ERβ-selective ligands from ERα ligands when treated with a panel of selective agonists and antagonists. Next, we tested an Environmental Protection Agency (EPA)-provided set of 45 estrogenic reference chemicals with known ERα in vivo activity and identified several that activated ERβ as well, with varying sensitivity, including a subset that is completely novel. We then used an orthogonal ERE-containing transgenic zebrafish (ZF) model to cross validate ERβ and ERα selective activities at the organism level. Using this environmentally relevant ZF assay, some compounds were confirmed to have ERβ activity, validating the GFP-ERβ:PRL-HeLa assay as a screening tool for potential ERβ active endocrine disruptors (EDCs). These data demonstrate the value of sensitive multiplex mechanistic data gathered by the GFP-ERβ:PRL-HeLa assay coupled with an orthogonal zebrafish model to rapidly identify environmentally relevant ERβ EDCs and improve upon currently available screening tools for this understudied nuclear receptor. |
| first_indexed | 2024-03-08T09:04:30Z |
| format | Article |
| id | doaj.art-72f160106c83423bac993c7ba9428a4b |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-03-08T09:04:30Z |
| publishDate | 2024-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-72f160106c83423bac993c7ba9428a4b2024-02-01T06:30:48ZengElsevierHeliyon2405-84402024-01-01101e23119A novel ERβ high throughput microscopy platform for testing endocrine disrupting chemicalsDerek A. Abbott0Maureen G. Mancini1Michael J. Bolt2Adam T. Szafran3Kaley A. Neugebauer4Fabio Stossi5Daniel A. Gorelick6Michael A. Mancini7Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USADepartment of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; GCC Center for Advanced Microscopy and Image Informatics, Houston, TX, USAGCC Center for Advanced Microscopy and Image Informatics, Houston, TX, USA; Center for Translational Cancer Research, Institute of Biosciences & Technology, Texas A&M University, Houston, TX, USADepartment of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; GCC Center for Advanced Microscopy and Image Informatics, Houston, TX, USACenter for Precision Environmental Health, Baylor College of Medicine, Houston, TX, USADepartment of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; GCC Center for Advanced Microscopy and Image Informatics, Houston, TX, USA; Corresponding author. GCC Center for Advanced Microscopy and Image Informatics, Department of Molecular and Cellular Biology, 117A Cullen Building, Baylor College of Medicine One Baylor Plaza Houston, TX 77030, USA.Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; Center for Precision Environmental Health, Baylor College of Medicine, Houston, TX, USADepartment of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA; GCC Center for Advanced Microscopy and Image Informatics, Houston, TX, USA; Center for Translational Cancer Research, Institute of Biosciences & Technology, Texas A&M University, Houston, TX, USA; Department of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, TX, USA; Corresponding author. GCC Center for Advanced Microscopy and Image Informatics Departments of Molecular and Cellular Biology, and Pharmacology and Chemical Biology 119A Cullen Building Baylor College of Medicine One Baylor Plaza Houston, TX 77030, 713, USA.In this study we present an inducible biosensor model for the Estrogen Receptor Beta (ERβ), GFP-ERβ:PRL-HeLa, a single-cell-based high throughput (HT) in vitro assay that allows direct visualization and measurement of GFP-tagged ERβ binding to ER-specific DNA response elements (EREs), ERβ-induced chromatin remodeling, and monitor transcriptional alterations via mRNA fluorescence in situ hybridization for a prolactin (PRL)-dsRED2 reporter gene. The model was used to accurately (Z’ = 0.58–0.8) differentiate ERβ-selective ligands from ERα ligands when treated with a panel of selective agonists and antagonists. Next, we tested an Environmental Protection Agency (EPA)-provided set of 45 estrogenic reference chemicals with known ERα in vivo activity and identified several that activated ERβ as well, with varying sensitivity, including a subset that is completely novel. We then used an orthogonal ERE-containing transgenic zebrafish (ZF) model to cross validate ERβ and ERα selective activities at the organism level. Using this environmentally relevant ZF assay, some compounds were confirmed to have ERβ activity, validating the GFP-ERβ:PRL-HeLa assay as a screening tool for potential ERβ active endocrine disruptors (EDCs). These data demonstrate the value of sensitive multiplex mechanistic data gathered by the GFP-ERβ:PRL-HeLa assay coupled with an orthogonal zebrafish model to rapidly identify environmentally relevant ERβ EDCs and improve upon currently available screening tools for this understudied nuclear receptor.http://www.sciencedirect.com/science/article/pii/S2405844023103276Estrogen receptor betaHigh content analysisHigh throughput microscopyEndocrine disruptorsZebrafishEstrogen receptor alpha |
| spellingShingle | Derek A. Abbott Maureen G. Mancini Michael J. Bolt Adam T. Szafran Kaley A. Neugebauer Fabio Stossi Daniel A. Gorelick Michael A. Mancini A novel ERβ high throughput microscopy platform for testing endocrine disrupting chemicals Heliyon Estrogen receptor beta High content analysis High throughput microscopy Endocrine disruptors Zebrafish Estrogen receptor alpha |
| title | A novel ERβ high throughput microscopy platform for testing endocrine disrupting chemicals |
| title_full | A novel ERβ high throughput microscopy platform for testing endocrine disrupting chemicals |
| title_fullStr | A novel ERβ high throughput microscopy platform for testing endocrine disrupting chemicals |
| title_full_unstemmed | A novel ERβ high throughput microscopy platform for testing endocrine disrupting chemicals |
| title_short | A novel ERβ high throughput microscopy platform for testing endocrine disrupting chemicals |
| title_sort | novel erβ high throughput microscopy platform for testing endocrine disrupting chemicals |
| topic | Estrogen receptor beta High content analysis High throughput microscopy Endocrine disruptors Zebrafish Estrogen receptor alpha |
| url | http://www.sciencedirect.com/science/article/pii/S2405844023103276 |
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