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Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials.

ABSTRACT: Engineered nanomaterials have been found to induce oxidative stress. Cellular oxidative stress, in turn, can result in the induction of antioxidant and detoxification enzymes which are controlled by the nuclear erythroid 2-related factor 2 (NRF2) transcription factor. Here, we present the results of a pre-validation study which was conducted within the frame of BIORIMA ("biomaterial risk management") an EU-funded research and innovation project. For this we used an NRF2 specific chemically activated luciferase expression reporter gene assay derived from the human U2OS osteosarcoma cell line to screen for the induction of the NRF2 mediated gene expression following exposure to biomedically relevant nanobiomaterials. Specifically, we investigated Fe₃O₄-PEG-PLGA nanomaterials while Ag and TiO₂ "benchmark" nanomaterials from the Joint Research Center were used as reference materials. The viability of the cells was determined by using the Alamar blue assay. We performed an interlaboratory study involving seven different laboratories to assess the applicability of the NRF2 reporter gene assay for the screening of nanobiomaterials. The latter work was preceded by online tutorials to ensure that the procedures were harmonized across the different participating laboratories. Fe₃O₄-PEG-PLGA nanomaterials were found to induce very limited NRF2 mediated gene expression, whereas exposure to Ag nanomaterials induced NRF2 mediated gene expression. TiO₂ nanomaterials did not induce NRF2 mediated gene expression. The variability in the results obtained by the participating laboratories was small with mean intra-laboratory standard deviation of 0.16 and mean inter laboratory standard deviation of 0.28 across all NRF2 reporter gene assay results. We conclude that the NRF2 reporter gene assay is a suitable assay for the screening of nanobiomaterial-induced oxidative stress responses.

SUBMITTER: Martin S

PROVIDER: S-EPMC9511406 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

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Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials.

Martin Sebastin S de Haan Laura L Miro Estruch Ignacio I Eder Kai Moritz KM Marzi Anne A Schnekenburger Jürgen J Blosi Magda M Costa Anna A Antonello Giulia G Bergamaschi Enrico E Riganti Chiara C Beal David D Carrière Marie M Taché Olivier O Hutchison Gary G Malone Eva E Young Lesley L Campagnolo Luisa L La Civita Fabio F Pietroiusti Antonio A Devineau Stéphanie S Baeza Armelle A Boland Sonja S Zong Cai C Ichihara Gaku G Fadeel Bengt B Bouwmeester Hans H

Frontiers in toxicology 20220905

Engineered nanomaterials have been found to induce oxidative stress. Cellular oxidative stress, in turn, can result in the induction of antioxidant and detoxification enzymes which are controlled by the nuclear erythroid 2-related factor 2 (NRF2) transcription factor. Here, we present the results of a pre-validation study which was conducted within the frame of BIORIMA ("biomaterial risk management") an EU-funded research and innovation project. For this we used an NRF2 specific chemically activ ...[more]

PMID: 36171865

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Project description:BackgroundHepatotoxicity accounts for a substantial number of drugs being withdrawn from the market. Using traditional animal models to detect hepatotoxicity is expensive and time-consuming. Alternative in vitro methods, in particular cell-based high-throughput screening (HTS) studies, have provided the research community with a large amount of data from toxicity assays. Among the various assays used to screen potential toxicants is the antioxidant response element beta lactamase reporter gene assay (ARE-bla), which identifies chemicals that have the potential to induce oxidative stress and was used to test > 10,000 compounds from the Tox21 program.ObjectiveThe ARE-bla computational model and HTS data from a big data source (PubChem) were used to profile environmental and pharmaceutical compounds with hepatotoxicity data.MethodsQuantitative structure-activity relationship (QSAR) models were developed based on ARE-bla data. The models predicted the potential oxidative stress response for known liver toxicants when no ARE-bla data were available. Liver toxicants were used as probe compounds to search PubChem Bioassay and generate a response profile, which contained thousands of bioassays (> 10 million data points). By ranking the in vitro-in vivo correlations (IVIVCs), the most relevant bioassay(s) related to hepatotoxicity were identified.ResultsThe liver toxicants profile contained the ARE-bla and relevant PubChem assays. Potential toxicophores for well-known toxicants were created by identifying chemical features that existed only in compounds with high IVIVCs.ConclusionProfiling chemical IVIVCs created an opportunity to fully explore the source-to-outcome continuum of modern experimental toxicology using cheminformatics approaches and big data sources.CitationKim MT, Huang R, Sedykh A, Wang W, Xia M, Zhu H. 2016. Mechanism profiling of hepatotoxicity caused by oxidative stress using antioxidant response element reporter gene assay models and big data. Environ Health Perspect 124:634-641;?http://dx.doi.org/10.1289/ehp.1509763.

Dataset Information

Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials.

Publications

Pre-validation of a reporter gene assay for oxidative stress for the rapid screening of nanobiomaterials.

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