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High-content imaging-based BAC-GFP toxicity pathway reporters to assess chemical adversity liabilities

Adaptive cellular stress responses are paramount in the healthy control of cell and tissue homeostasis and generally activated during toxicity in a chemical-specific manner. Here, we established a platform containing a panel of distinct adaptive stress response reporter cell lines based on BAC-trans...

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Autores principales: Wink, Steven, Hiemstra, Steven, Herpers, Bram, van de Water, Bob
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5316409/
https://www.ncbi.nlm.nih.gov/pubmed/27358234
http://dx.doi.org/10.1007/s00204-016-1781-0
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author Wink, Steven
Hiemstra, Steven
Herpers, Bram
van de Water, Bob
author_facet Wink, Steven
Hiemstra, Steven
Herpers, Bram
van de Water, Bob
author_sort Wink, Steven
collection PubMed
description Adaptive cellular stress responses are paramount in the healthy control of cell and tissue homeostasis and generally activated during toxicity in a chemical-specific manner. Here, we established a platform containing a panel of distinct adaptive stress response reporter cell lines based on BAC-transgenomics GFP tagging in HepG2 cells. Our current panel of eleven BAC-GFP HepG2 reporters together contains (1) upstream sensors, (2) downstream transcription factors and (3) their respective target genes, representing the oxidative stress response pathway (Keap1/Nrf2/Srxn1), the unfolded protein response in the endoplasmic reticulum (Xbp1/Atf4/BiP/Chop) and the DNA damage response (53bp1/p53/p21). Using automated confocal imaging and quantitative single-cell image analysis, we established that all reporters allowed the time-resolved, sensitive and mode-of-action-specific activation of the individual BAC-GFP reporter cell lines as defined by a panel of pathway-specific training compounds. Implementing the temporal pathway activity information increased the discrimination of training compounds. For a set of >30 hepatotoxicants, the induction of Srxn1, BiP, Chop and p21 BAC-GFP reporters correlated strongly with the transcriptional responses observed in cryopreserved primary human hepatocytes. Together, our data indicate that a phenotypic adaptive stress response profiling platform will allow a high throughput and time-resolved classification of chemical-induced stress responses, thus assisting in the future mechanism-based safety assessment of chemicals. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00204-016-1781-0) contains supplementary material, which is available to authorized users.
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spelling pubmed-53164092017-03-03 High-content imaging-based BAC-GFP toxicity pathway reporters to assess chemical adversity liabilities Wink, Steven Hiemstra, Steven Herpers, Bram van de Water, Bob Arch Toxicol In vitro Systems Adaptive cellular stress responses are paramount in the healthy control of cell and tissue homeostasis and generally activated during toxicity in a chemical-specific manner. Here, we established a platform containing a panel of distinct adaptive stress response reporter cell lines based on BAC-transgenomics GFP tagging in HepG2 cells. Our current panel of eleven BAC-GFP HepG2 reporters together contains (1) upstream sensors, (2) downstream transcription factors and (3) their respective target genes, representing the oxidative stress response pathway (Keap1/Nrf2/Srxn1), the unfolded protein response in the endoplasmic reticulum (Xbp1/Atf4/BiP/Chop) and the DNA damage response (53bp1/p53/p21). Using automated confocal imaging and quantitative single-cell image analysis, we established that all reporters allowed the time-resolved, sensitive and mode-of-action-specific activation of the individual BAC-GFP reporter cell lines as defined by a panel of pathway-specific training compounds. Implementing the temporal pathway activity information increased the discrimination of training compounds. For a set of >30 hepatotoxicants, the induction of Srxn1, BiP, Chop and p21 BAC-GFP reporters correlated strongly with the transcriptional responses observed in cryopreserved primary human hepatocytes. Together, our data indicate that a phenotypic adaptive stress response profiling platform will allow a high throughput and time-resolved classification of chemical-induced stress responses, thus assisting in the future mechanism-based safety assessment of chemicals. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00204-016-1781-0) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2016-06-29 2017 /pmc/articles/PMC5316409/ /pubmed/27358234 http://dx.doi.org/10.1007/s00204-016-1781-0 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle In vitro Systems
Wink, Steven
Hiemstra, Steven
Herpers, Bram
van de Water, Bob
High-content imaging-based BAC-GFP toxicity pathway reporters to assess chemical adversity liabilities
title High-content imaging-based BAC-GFP toxicity pathway reporters to assess chemical adversity liabilities
title_full High-content imaging-based BAC-GFP toxicity pathway reporters to assess chemical adversity liabilities
title_fullStr High-content imaging-based BAC-GFP toxicity pathway reporters to assess chemical adversity liabilities
title_full_unstemmed High-content imaging-based BAC-GFP toxicity pathway reporters to assess chemical adversity liabilities
title_short High-content imaging-based BAC-GFP toxicity pathway reporters to assess chemical adversity liabilities
title_sort high-content imaging-based bac-gfp toxicity pathway reporters to assess chemical adversity liabilities
topic In vitro Systems
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5316409/
https://www.ncbi.nlm.nih.gov/pubmed/27358234
http://dx.doi.org/10.1007/s00204-016-1781-0
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