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Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models

The evaluation of exposure to multiple contaminants in a mixture presents a number of challenges. For example, the characterization of chemical metabolism in a mixture setting remains a research area with critical knowledge gaps. Studies of chemical metabolism typically utilize suspension cultures o...

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Autores principales: Valdiviezo, Alan, Kato, Yuki, Baker, Erin S., Chiu, Weihsueh A., Rusyn, Ivan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9609317/
https://www.ncbi.nlm.nih.gov/pubmed/36287846
http://dx.doi.org/10.3390/toxics10100566
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author Valdiviezo, Alan
Kato, Yuki
Baker, Erin S.
Chiu, Weihsueh A.
Rusyn, Ivan
author_facet Valdiviezo, Alan
Kato, Yuki
Baker, Erin S.
Chiu, Weihsueh A.
Rusyn, Ivan
author_sort Valdiviezo, Alan
collection PubMed
description The evaluation of exposure to multiple contaminants in a mixture presents a number of challenges. For example, the characterization of chemical metabolism in a mixture setting remains a research area with critical knowledge gaps. Studies of chemical metabolism typically utilize suspension cultures of primary human hepatocytes; however, this model is not suitable for studies of more extended exposures and donor-to-donor variability in a metabolic capacity is unavoidable. To address this issue, we utilized several in vitro models based on human-induced pluripotent stem cell (iPSC)-derived hepatocytes (iHep) to characterize the metabolism of an equimolar (1 or 5 µM) mixture of 20 pesticides. We used iHep suspensions and 2D sandwich cultures, and a microphysiological system OrganoPlate(®) 2-lane 96 (Mimetas(TM)) that also included endothelial cells and THP-1 cell-derived macrophages. When cell culture media were evaluated using gas and liquid chromatography coupled to tandem mass spectrometry methods, we found that the parent molecule concentrations diminished, consistent with metabolic activity. This effect was most pronounced in iHep suspensions with a 1 µM mixture, and was lowest in OrganoPlate(®) 2-lane 96 for both mixtures. Additionally, we used ion mobility spectrometry–mass spectrometry (IMS-MS) to screen for metabolite formation in these cultures. These analyses revealed the presence of five primary metabolites that allowed for a more comprehensive evaluation of chemical metabolism in vitro. These findings suggest that iHep-based suspension assays maintain higher metabolic activity compared to 2D sandwich and OrganoPlate(®) 2-lane 96 model. Moreover, this study illustrates that IMS-MS can characterize in vitro metabolite formation following exposure to mixtures of environmental contaminants.
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spelling pubmed-96093172022-10-28 Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models Valdiviezo, Alan Kato, Yuki Baker, Erin S. Chiu, Weihsueh A. Rusyn, Ivan Toxics Article The evaluation of exposure to multiple contaminants in a mixture presents a number of challenges. For example, the characterization of chemical metabolism in a mixture setting remains a research area with critical knowledge gaps. Studies of chemical metabolism typically utilize suspension cultures of primary human hepatocytes; however, this model is not suitable for studies of more extended exposures and donor-to-donor variability in a metabolic capacity is unavoidable. To address this issue, we utilized several in vitro models based on human-induced pluripotent stem cell (iPSC)-derived hepatocytes (iHep) to characterize the metabolism of an equimolar (1 or 5 µM) mixture of 20 pesticides. We used iHep suspensions and 2D sandwich cultures, and a microphysiological system OrganoPlate(®) 2-lane 96 (Mimetas(TM)) that also included endothelial cells and THP-1 cell-derived macrophages. When cell culture media were evaluated using gas and liquid chromatography coupled to tandem mass spectrometry methods, we found that the parent molecule concentrations diminished, consistent with metabolic activity. This effect was most pronounced in iHep suspensions with a 1 µM mixture, and was lowest in OrganoPlate(®) 2-lane 96 for both mixtures. Additionally, we used ion mobility spectrometry–mass spectrometry (IMS-MS) to screen for metabolite formation in these cultures. These analyses revealed the presence of five primary metabolites that allowed for a more comprehensive evaluation of chemical metabolism in vitro. These findings suggest that iHep-based suspension assays maintain higher metabolic activity compared to 2D sandwich and OrganoPlate(®) 2-lane 96 model. Moreover, this study illustrates that IMS-MS can characterize in vitro metabolite formation following exposure to mixtures of environmental contaminants. MDPI 2022-09-27 /pmc/articles/PMC9609317/ /pubmed/36287846 http://dx.doi.org/10.3390/toxics10100566 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Valdiviezo, Alan
Kato, Yuki
Baker, Erin S.
Chiu, Weihsueh A.
Rusyn, Ivan
Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models
title Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models
title_full Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models
title_fullStr Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models
title_full_unstemmed Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models
title_short Evaluation of Metabolism of a Defined Pesticide Mixture through Multiple In Vitro Liver Models
title_sort evaluation of metabolism of a defined pesticide mixture through multiple in vitro liver models
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9609317/
https://www.ncbi.nlm.nih.gov/pubmed/36287846
http://dx.doi.org/10.3390/toxics10100566
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