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Article ; Online: Constructing xenobiotic maps of metabolism to predict enzymes catalyzing metabolites capable of binding to DNA.

Conan, Mael / Théret, Nathalie / Langouet, Sophie / Siegel, Anne

2021 Volume 22, Issue 1, Page(s) 450

Abstract: Background: The liver plays a major role in the metabolic activation of xenobiotics (drugs, chemicals such as pollutants, pesticides, food additives ... ). Among environmental contaminants of concern, heterocyclic aromatic amines (HAA) are xenobiotics ... ...

Abstract	Background: The liver plays a major role in the metabolic activation of xenobiotics (drugs, chemicals such as pollutants, pesticides, food additives...). Among environmental contaminants of concern, heterocyclic aromatic amines (HAA) are xenobiotics classified by IARC as possible or probable carcinogens (2A or 2B). There exist little information about the effect of these HAA in humans. While HAA is a family of more than thirty identified chemicals, the metabolic activation and possible DNA adduct formation have been fully characterized in human liver for only a few of them (MeIQx, PhIP, A[Formula: see text]C). Results: We have developed a modeling approach in order to predict all the possible metabolites of a xenobiotic and enzymatic profiles that are linked to the production of metabolites able to bind DNA. Our prediction of metabolites approach relies on the construction of an enriched and annotated map of metabolites from an input metabolite.The pipeline assembles reaction prediction tools (SyGMa), sites of metabolism prediction tools (Way2Drug, SOMP and Fame 3), a tool to estimate the ability of a xenobotics to form DNA adducts (XenoSite Reactivity V1), and a filtering procedure based on Bayesian framework. This prediction pipeline was evaluated using caffeine and then applied to HAA. The method was applied to determine enzymes profiles associated with the maximization of metabolites derived from each HAA which are able to bind to DNA. The classification of HAA according to enzymatic profiles was consistent with their chemical structures. Conclusions: Overall, a predictive toxicological model based on an in silico systems biology approach opens perspectives to estimate the genotoxicity of various chemical classes of environmental contaminants. Moreover, our approach based on enzymes profile determination opens the possibility of predicting various xenobiotics metabolites susceptible to bind to DNA in both normal and physiopathological situations.
MeSH term(s)	Amines ; Bayes Theorem ; Carcinogens ; DNA Adducts ; Humans ; Xenobiotics
Chemical Substances	Amines ; Carcinogens ; DNA Adducts ; Xenobiotics
Language	English
Publishing date	2021-09-21
Publishing country	England
Document type	Journal Article
ZDB-ID	2041484-5
ISSN	1471-2105 ; 1471-2105
ISSN (online)	1471-2105
ISSN	1471-2105
DOI	10.1186/s12859-021-04363-6
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Article ; Online: Hepatotoxicity of cyanotoxin microcystin-LR in human: Insights into mechanisms of action in the 3D culture model Hepoid-HepaRG.

Chowdhury, Riju R / Rose, Sophie / Ezan, Frédéric / Sovadinová, Iva / Babica, Pavel / Langouët, Sophie

Environmental pollution (Barking, Essex : 1987)

2023 Volume 342, Page(s) 123047

Abstract: Microcystin-LR (MC-LR) is a potent hepatotoxin produced by harmful cyanobacterial blooms (CyanoHABs). MC-LR targets highly differentiated hepatocytes expressing organic anion transporting polypeptides OATP1B1 and OATP1B3 that are responsible for ... ...

Abstract	Microcystin-LR (MC-LR) is a potent hepatotoxin produced by harmful cyanobacterial blooms (CyanoHABs). MC-LR targets highly differentiated hepatocytes expressing organic anion transporting polypeptides OATP1B1 and OATP1B3 that are responsible for hepatocellular uptake of the toxin. The present study utilized an advanced 3D in vitro human liver model Hepoid-HepaRG based on the cultivation of collagen-matrix embedded multicellular spheroids composed of highly differentiated and polarized hepatocyte-like cells. 14-d-old Hepoid-HepaRG cultures showed increased expression of OATP1B1/1B3 and sensitivity to MC-LR cytotoxicity at concentrations >10 nM (48 h exposure, EC
MeSH term(s)	Humans ; Carcinoma, Hepatocellular ; Liver Neoplasms ; Microcystins/toxicity ; Microcystins/metabolism ; Cadherins ; Chemical and Drug Induced Liver Injury ; Marine Toxins
Chemical Substances	cyanoginosin LR (EQ8332842Y) ; Microcystins ; Cadherins ; Marine Toxins
Language	English
Publishing date	2023-11-29
Publishing country	England
Document type	Journal Article
ZDB-ID	280652-6
ISSN	1873-6424 ; 0013-9327 ; 0269-7491
ISSN (online)	1873-6424
ISSN	0013-9327 ; 0269-7491
DOI	10.1016/j.envpol.2023.123047
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Article ; Online: ReadEDTest: A tool to assess the readiness of in vitro test methods under development for identifying endocrine disruptors

Crouzet, Thibault / Grignard, Elise / Brion, François / Blanc, Etienne B. / Podechard, Normand / Langouet, Sophie / Alonso-Magdalena, Paloma / Hubert, Philippe / Kim, Min-ji / Audouze, Karine

Environment International. 2023 Apr., v. 174 p.107910-

2023

Abstract: Growing evidence shows that endocrine disruptors (EDs), known to affect the reproductive system, may also disturb other hormone-regulated functions leading to cancers, neurodevelopmental defects, metabolic and immune diseases. To reduce exposure to EDs ... ...

Abstract	Growing evidence shows that endocrine disruptors (EDs), known to affect the reproductive system, may also disturb other hormone-regulated functions leading to cancers, neurodevelopmental defects, metabolic and immune diseases. To reduce exposure to EDs and limit their health effects, development of screening and mechanism-based assays to identify EDs is encouraged. Nevertheless, the crucial validation step of test methods by regulatory bodies is a time- and resource-consuming process. One of the main raisons of this long duration process is that method developers, mainly researchers, are not fully aware of the regulatory needs to validate a test. We propose an online self-assessment questionnaire (SAQ) called ReadEDTest easy to be used by all researchers. The aim of ReadEDTest is to speed up the validation process by assessing readiness criteria of in vitro and fish embryo ED test methods under development. The SAQ is divided into 7 sections and 13 sub-sections containing essential information requested by the validating bodies. The readiness of the tests can be assessed by specific score limits for each sub-section. Results are displayed via a graphical representation to help identification of the sub-sections having sufficient or insufficient information. The relevance of the proposed innovative tool was supported using two test methods already validated by the OECD and four under development test methods.
Keywords	environment ; fish ; questionnaires ; reproductive system ; Endocrine disrupting chemicals ; Zebrafish embryo ; OBERON ; Pepper ; Validation process ; IATA ; OECD
Language	English
Dates of publication	2023-04
Publishing place	Elsevier Ltd
Document type	Article ; Online
ZDB-ID	554791-x
ISSN	1873-6750 ; 0160-4120
ISSN (online)	1873-6750
ISSN	0160-4120
DOI	10.1016/j.envint.2023.107910
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Generation of proliferating human adult hepatocytes using optimized 3D culture conditions.

Rose, Sophie / Ezan, Frédéric / Cuvellier, Marie / Bruyère, Arnaud / Legagneux, Vincent / Langouët, Sophie / Baffet, Georges

Scientific reports

2021 Volume 11, Issue 1, Page(s) 515

Abstract: Generating the proliferation of differentiated normal adult human hepatocytes is a major challenge and an expected central step in understanding the microenvironmental conditions that regulate the phenotype of human hepatocytes in vitro. In this work, we ...

Abstract	Generating the proliferation of differentiated normal adult human hepatocytes is a major challenge and an expected central step in understanding the microenvironmental conditions that regulate the phenotype of human hepatocytes in vitro. In this work, we described optimized 3D culture conditions of primary human hepatocytes (PHH) to trigger two waves of proliferation and we identified matrix stiffness and cell-cell interactions as the main actors necessary for this proliferation. We demonstrated that DNA replication and overexpression of cell cycle markers are modulate by the matrix stiffness while PHH cultured in 3D without prior cellular interactions did not proliferate. Besides, we showed that PHH carry out an additional cell cycle after transient inhibition of MAPK MER1/2-ERK1/2 signaling pathway. Collagen cultured hepatocytes are organized as characteristic hollow spheroids able to maintain survival, cell polarity and hepatic differentiation for long-term culture periods of at least 28 days. Remarkably, we demonstrated by transcriptomic analysis and functional experiments that proliferating cells are mature hepatocytes with high detoxication capacities. In conclusion, the advanced 3D model described here, named Hepoid, is particularly relevant for obtaining normal human proliferating hepatocytes. By allowing concomitant proliferation and differentiation, it constitutes a promising tool for many pharmacological and biotechnological applications.
MeSH term(s)	Cell Communication ; Cell Culture Techniques/methods ; Cell Cycle ; Cell Differentiation ; Cell Polarity ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Collagen ; DNA Replication ; Elasticity ; Hepatocytes/physiology ; Humans ; MAP Kinase Signaling System ; Spheroids, Cellular
Chemical Substances	Collagen (9007-34-5)
Language	English
Publishing date	2021-01-12
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-020-80019-4
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Article ; Online: In silico prediction of Heterocyclic Aromatic Amines metabolism susceptible to form DNA adducts in humans.

Delannée, Victorien / Langouët, Sophie / Siegel, Anne / Théret, Nathalie

Toxicology letters

2018 Volume 300, Page(s) 18–30

Abstract: Heterocyclic Aromatic Amines (HAAs) are environmental and food contaminants that are classified as probable or possible carcinogens by the International Agency for Research on Cancer. Thirty different HAAs have been identified. However the metabolism of ... ...

Abstract	Heterocyclic Aromatic Amines (HAAs) are environmental and food contaminants that are classified as probable or possible carcinogens by the International Agency for Research on Cancer. Thirty different HAAs have been identified. However the metabolism of only three of them have been fully characterized in human hepatocytes: AαC (2-amino-9H-pyrido[2,3-b]indole), MeIQx (2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline) and PhIP (2-amino-1-methyl-6-phenyl-imidazo[4,5-b]pyridine). In this study, we use an integrative approach to accurately predict the biotransformation of 30 HAAs into DNA reactive and non DNA reactive compounds. We first build predicted metabolites networks by iterating a knowledge-based expert system of prediction of metabolic reactions based on fingerprint similarities. Next, we combine several methods for predicting Sites Of Metabolism (SOM) in order to reduce the metabolite reaction graphs and to predict the metabolites reactive with DNA. We validate the method by comparing the experimental versus predicted data for the known AαC, MeIQx and PhIP metabolism. 28 of the 30 experimentally determined metabolites are well predicted and 9 of the 10 metabolites known to form DNA adducts are predicted with a high probability to be reactive with DNA. Applying our approach to the 27 unknown HAAs, we generate maps for the metabolic biotransformation of each HAA, including new metabolites with a high-predicted DNA reactivity, which can be further explored through an user-friendly and interactive web interface.
MeSH term(s)	Amines/chemistry ; Amines/metabolism ; Carcinogens/chemistry ; Carcinogens/metabolism ; DNA Adducts/metabolism ; Hepatocytes/metabolism ; Heterocyclic Compounds/chemistry ; Heterocyclic Compounds/metabolism ; Humans
Chemical Substances	Amines ; Carcinogens ; DNA Adducts ; Heterocyclic Compounds
Language	English
Publishing date	2018-10-10
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	433788-8
ISSN	1879-3169 ; 0378-4274
ISSN (online)	1879-3169
ISSN	0378-4274
DOI	10.1016/j.toxlet.2018.10.011
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Article: A modeling approach to evaluate the balance between bioactivation and detoxification of MeIQx in human hepatocytes.

Delannée, Victorien / Langouët, Sophie / Théret, Nathalie / Siegel, Anne

PeerJ

2017 Volume 5, Page(s) e3703

Abstract: Background: Heterocyclic aromatic amines (HAA) are environmental and food contaminants that are potentially carcinogenic for humans. 2-Amino-3,8-dimethylimidazo[4,5-: Methods: Using a computational approach, we developed a numerical model for MeIQx ... ...

Abstract	Background: Heterocyclic aromatic amines (HAA) are environmental and food contaminants that are potentially carcinogenic for humans. 2-Amino-3,8-dimethylimidazo[4,5- Methods: Using a computational approach, we developed a numerical model for MeIQx metabolism in the liver that predicts the MeIQx biotransformation into detoxification or bioactivation pathways according to the concentration of MeIQx. Results: Our results demonstrate that (1) the detoxification pathway predominates, (2) the ratio between detoxification and bioactivation pathways is not linear and shows a maximum at 10 µM of MeIQx in hepatocyte cell models, and (3) CYP1A2 is a key enzyme in the system that regulates the balance between bioactivation and detoxification. Our analysis suggests that such a ratio could be considered as an indicator of MeIQx genotoxicity at a low concentration of MeIQx. Conclusions: Our model permits the investigation of the balance between bioactivation (i.e., DNA adduct formation pathway through the prediction of potential genotoxic compounds) and detoxification of MeIQx in order to predict the behaviour of this environmental contaminant in the human liver. It highlights the importance of complex regulations of enzyme competitions that should be taken into account in any further multi-organ models.
Language	English
Publishing date	2017-09-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	2703241-3
ISSN	2167-8359
ISSN	2167-8359
DOI	10.7717/peerj.3703
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Article ; Online: Assessment of endocrine disruptor impacts on lipid metabolism in a fatty acid-supplemented HepaRG human hepatic cell line.

Bernal, Kévin / Touma, Charbel / Le-Grand, Béatrice / Rose, Sophie / Degerli, Selenay / Genêt, Valentine / Lagadic-Gossmann, Dominique / Coumoul, Xavier / Martin-Chouly, Corinne / Langouët, Sophie / Blanc, Etienne B

Chemosphere

2023 Volume 349, Page(s) 140883

Abstract: The incidence of metabolic dysfunction-associated steatotic liver disease (MASLD) is increasing worldwide. This disease encompasses several stages, from steatosis to steatohepatitis and, eventually, to fibrosis and cirrhosis. Exposure to environmental ... ...

Abstract	The incidence of metabolic dysfunction-associated steatotic liver disease (MASLD) is increasing worldwide. This disease encompasses several stages, from steatosis to steatohepatitis and, eventually, to fibrosis and cirrhosis. Exposure to environmental contaminants is one of the risk factors and an increasing amount of evidence points to a role for endocrine disrupting compounds (EDCs). This study assesses the impact of selected EDCs on the formation of lipid droplets, the marker for steatosis in a hepatic model. The mechanisms underlying this effect are then explored. Ten compounds were selected according to their obesogenic properties: bisphenol A, F and S, butyl-paraben, cadmium chloride, p,p'-DDE, DBP, DEHP, PFOA and PFOS. Using a 2D or 3D model, HepaRG cells were exposed to the compounds with or without fatty acid supplementation. Then, the formation of lipid droplets was quantified by an automated fluorescence-based method. The expression of genes and proteins involved in lipid metabolism and the impact on cellular respiration was analyzed. The formation of lipid droplets, which is revealed or enhanced by oleic acid supplementation, was most effectively induced by p,p'-DDE and DEHP. Experiments employing either 2D or 3D culture conditions gave similar results. Both compounds induced the expression of PLIN2. p,p'-DDE also appears to act by decreasing in fatty acid oxidation. Some EDCs were able to induce the formation of lipid droplets, in HepaRG cells, an effect which was increased after supplementation of the cells with oleic acid. A full understanding of the mechanisms of these effects will require further investigation. The novel automated detection method described here may also be useful in the future as a regulatory test for EDC risk assessment.
MeSH term(s)	Humans ; Lipid Metabolism ; Fatty Acids/metabolism ; Endocrine Disruptors/metabolism ; Oleic Acid/toxicity ; Oleic Acid/metabolism ; Dichlorodiphenyl Dichloroethylene/metabolism ; Diethylhexyl Phthalate/toxicity ; Fatty Liver/metabolism ; Hepatocytes
Chemical Substances	Fatty Acids ; Endocrine Disruptors ; Oleic Acid (2UMI9U37CP) ; Dichlorodiphenyl Dichloroethylene (4M7FS82U08) ; Diethylhexyl Phthalate (C42K0PH13C)
Language	English
Publishing date	2023-12-11
Publishing country	England
Document type	Journal Article
ZDB-ID	120089-6
ISSN	1879-1298 ; 0045-6535 ; 0366-7111
ISSN (online)	1879-1298
ISSN	0045-6535 ; 0366-7111
DOI	10.1016/j.chemosphere.2023.140883
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Article ; Online: ReadEDTest: A tool to assess the readiness of in vitro test methods under development for identifying endocrine disruptors.

Crouzet, Thibault / Grignard, Elise / Brion, François / Blanc, Etienne B / Podechard, Normand / Langouet, Sophie / Alonso-Magdalena, Paloma / Hubert, Philippe / Kim, Min Ji / Audouze, Karine

Environment international

2023 Volume 174, Page(s) 107910

Abstract	Growing evidence shows that endocrine disruptors (EDs), known to affect the reproductive system, may also disturb other hormone-regulated functions leading to cancers, neurodevelopmental defects, metabolic and immune diseases. To reduce exposure to EDs and limit their health effects, development of screening and mechanism-based assays to identify EDs is encouraged. Nevertheless, the crucial validation step of test methods by regulatory bodies is a time- and resource-consuming process. One of the main raisons of this long duration process is that method developers, mainly researchers, are not fully aware of the regulatory needs to validate a test. We propose an online self-assessment questionnaire (SAQ) called ReadEDTest easy to be used by all researchers. The aim of ReadEDTest is to speed up the validation process by assessing readiness criteria of in vitro and fish embryo ED test methods under development. The SAQ is divided into 7 sections and 13 sub-sections containing essential information requested by the validating bodies. The readiness of the tests can be assessed by specific score limits for each sub-section. Results are displayed via a graphical representation to help identification of the sub-sections having sufficient or insufficient information. The relevance of the proposed innovative tool was supported using two test methods already validated by the OECD and four under development test methods.
MeSH term(s)	Animals ; Endocrine Disruptors/toxicity ; Endocrine Disruptors/metabolism ; In Vitro Techniques
Chemical Substances	Endocrine Disruptors
Language	English
Publishing date	2023-03-31
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	554791-x
ISSN	1873-6750 ; 0160-4120
ISSN (online)	1873-6750
ISSN	0160-4120
DOI	10.1016/j.envint.2023.107910
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Article ; Online: DMSO-free highly differentiated HepaRG spheroids for chronic toxicity, liver functions and genotoxicity studies.

Rose, Sophie / Cuvellier, Marie / Ezan, Frédéric / Carteret, Jennifer / Bruyère, Arnaud / Legagneux, Vincent / Nesslany, Fabrice / Baffet, Georges / Langouët, Sophie

Archives of toxicology

2021 Volume 96, Issue 1, Page(s) 243–258

Abstract: The liver is essential in the elimination of environmental and food contaminants. Given the interspecies differences between rodents and humans, the development of relevant in vitro human models is crucial to investigate liver functions and toxicity in ... ...

Abstract	The liver is essential in the elimination of environmental and food contaminants. Given the interspecies differences between rodents and humans, the development of relevant in vitro human models is crucial to investigate liver functions and toxicity in cells that better reflect pathophysiological processes. Classically, the differentiation of the hepatic HepaRG cell line requires high concentration of dimethyl sulfoxide (DMSO), which restricts its usefulness for drug-metabolism studies. Herein, we describe undifferentiated HepaRG cells embedded in a collagen matrix in DMSO-free conditions that rapidly organize into polarized hollow spheroids of differentiated hepatocyte-like cells (Hepoid-HepaRG). Our conditions allow concomitant proliferation with high levels of liver-specific functions and xenobiotic metabolism enzymes expression and activities after a few days of culture and for at least 4 weeks. By studying the toxicity of well-known injury-inducing drugs by treating cells with 1- to 100-fold of their plasmatic concentrations, we showed appropriate responses and demonstrate the sensitivity to drugs known to induce various degrees of liver injury. Our results also demonstrated that the model is well suited to estimate cholestasis and steatosis effects of drugs following chronic treatment. Additionally, DNA alterations caused by four genotoxic compounds (Aflatoxin B
MeSH term(s)	DNA Damage ; Dimethyl Sulfoxide/toxicity ; Hepatocytes ; Liver ; Micronucleus Tests/methods
Chemical Substances	Dimethyl Sulfoxide (YOW8V9698H)
Language	English
Publishing date	2021-11-11
Publishing country	Germany
Document type	Journal Article
ZDB-ID	124992-7
ISSN	1432-0738 ; 0340-5761
ISSN (online)	1432-0738
ISSN	0340-5761
DOI	10.1007/s00204-021-03178-x
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Article ; Online: In vitro

Cuvellier, Marie / Rose, Sophie / Ezan, Frédéric / Jarry, Ulrich / de Oliveira, Hugo / Bruyère, Arnaud / Drieu La Rochelle, Christophe / Legagneux, Vincent / Langouët, Sophie / Baffet, Georges

Biofabrication

2022 Volume 14, Issue 3

Abstract: In recent decades, ... ...

Abstract	In recent decades, 3D
MeSH term(s)	Animals ; Bioprinting/methods ; Gelatin/chemistry ; Hepatocytes/metabolism ; Humans ; Hydrogels/chemistry ; Mice ; Printing, Three-Dimensional ; Tissue Engineering/methods ; Tissue Scaffolds/chemistry
Chemical Substances	Hydrogels ; Gelatin (9000-70-8)
Language	English
Publishing date	2022-06-30
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2500944-8
ISSN	1758-5090 ; 1758-5082
ISSN (online)	1758-5090
ISSN	1758-5082
DOI	10.1088/1758-5090/ac7825
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