Quantitative interpretation of <scp>ToxTracker</scp> dose–response data for potency comparisons and <scp>mode‐of‐action</scp> determination

Boisvert, Lorrie; Derr, Remco S.; Østerlund, Torben; Hendriks, Giel; Brandsma, Inger

doi:10.1002/em.22525

Cited by 6 publications

(4 citation statements)

References 43 publications

(60 reference statements)

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“…ToxTracker uses a panel containing six green fluorescent reporter proteins, one for each cell line, to evaluate the ability of genotoxic agents to react with genetic material. The result of this interaction may be blocking of DNA replication, induction of oxidative stress, activation of reactions mediated by denaturation of proteins or general reactions of cellular stress, dependent on the transcription factor R5, which serves as a suppressor of tumor growth [212,213]. The ToxTracker system allows determining the genotoxic potential of the test compound in one test, taking into account various endpoints of DNA damage [213].…”

Section: Modern Methods For Assessing the Genotoxicity Of Environment...mentioning

confidence: 99%

Systems for Genetic Assessment of the Impact of Environmental Factors

Kislyak,

Dugan,

Yalovenko

2024

Innov Biosyst Bioeng

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One of the most important components of environmental protection is the development of hygiene standards aimed at shielding the human population from the adverse effects of environmental pollution. The European and American Chemical Societies have reported approximately 800,000 chemicals, with no available information on potential risks to human genetic health and negative environmental impact. Given the exponential increase in chemical compounds generated by humanity in various industries, the issue of effectivly identifying and accounting for various genetic and carcinogenic hazards is particularle relevant. The assessment of potential genotoxicity of environmental factors is an integral part of genetic safety assessment for both prokaryotic and eukaryotic organisms, including humans. The evaluation of the genetic activity of chemical compounds is a fundamentsl requirement for their comprehensive toxicological assessment. From the perspective of genetic and epigenetic mechanisms of influence, our review considers standard methods for detecting and assessing the potential genetic hazard associated with environmental factors. These methods are part of a standard, generally accepted test system battery. Additionally, the review covers some modern experimental methods that are not widely accepted today. A detailed analysis of approaches to the assessment of potential genetic mutagenic activity was carried out, presenting their main advantages and disadvantages. Taking into account the recommendations issued by the Organisation for Economic Co-operation and Development on testing hazardous chemical compounds that may affect human health, an attempt was made to find optimal approaches to solving the task of predicting genetic effects and their consequences for humans.

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Section: Modern Methods For Assessing the Genotoxicity Of Environment...mentioning

confidence: 99%

Systems for Genetic Assessment of the Impact of Environmental Factors

Kislyak,

Dugan,

Yalovenko

2024

Innov Biosyst Bioeng

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“…This twofold increase as cut-off for a positive response is based on three times the standard deviation (SD) of fluorescence levels in solvent control cultures. The validity of this approach was recently confirmed using a bootstrapping analysis of more than a 1000 vehicle control treated cultures (Boisvert et al, 2023). Compounds were classified as nongenotoxic in ToxTracker if the induction levels of the genotoxicity reporters (Bscl2 and Rtkn) were less than 1.5-fold.…”

Section: Toxtracker Prediction Modelmentioning

confidence: 98%

“…The criteria for a positive or negative result for each GFP-reporter, were previously established (Boisvert et al, 2023;Hendriks et al, 2016) and are defined in Table 1. GFP reporter inductions at concentrations inducing ≤75% cytotoxicity were acceptable for Tox-Tracker analysis.…”

Section: Toxtracker Prediction Modelmentioning

confidence: 99%

Interlaboratory validation of the ToxTracker assay: An in vitro reporter assay for mechanistic genotoxicity assessment

Hendriks,

Adriaens,

Allemang

et al. 2024

Environ and Mol Mutagen

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ToxTracker is a mammalian cell reporter assay that predicts the genotoxic properties of compounds with high accuracy. By evaluating induction of various reporter genes that play a key role in relevant cellular pathways, it provides insight into chemical mode‐of‐action (MoA), thereby supporting discrimination of direct‐acting genotoxicants and cytotoxic chemicals. A comprehensive interlaboratory validation trial was conducted, in which the principles outlined in OECD Guidance Document 34 were followed, with the primary objectives of establishing transferability and reproducibility of the assay and confirming the ability of ToxTracker to correctly classify genotoxic and non‐genotoxic compounds. Reproducibility of the assay to predict genotoxic MoA was confirmed across participating laboratories and data were evaluated in terms of concordance with in vivo genotoxicity outcomes. Seven laboratories tested a total of 64 genotoxic and non‐genotoxic chemicals that together cover a broad chemical space. The within‐laboratory reproducibility (WLR) was up to 98% (73%–98% across participants) and the overall between‐laboratory reproducibility (BLR) was 83%. This trial confirmed the accuracy of ToxTracker to predict in vivo genotoxicants with a sensitivity of 84.4% and a specificity of 91.2%. We concluded that ToxTracker is a robust in vitro assay for the accurate prediction of in vivo genotoxicity. Considering ToxTracker's robust standalone accuracy and that it can provide important information on the MoA of chemicals, it is seen as a valuable addition to the regulatory in vitro genotoxicity battery that may even have the potential to replace certain currently used in vitro battery assays.

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“…The detection of GFP fluorescence in the Bscl2-GFP and Rtkn-GFP cell lines is indicative of DNA replication inhibition and DNA damage, respectively (Hendriks et al, 2016). The ToxTracker assay has been applied to assess over 120 chemicals with different MOAs and quantitative analyses have demonstrated that ToxTracker-informed potency determinations are more informative than simply using the data qualitatively for hazard identification (Boisvert et al, 2023;Wills et al, 2021a). Extended validation studies of the ToxTracker assay are currently being conducted and the information will be used to draft a detailed review paper in support of TG development.…”

Section: Indicator Assaysmentioning

confidence: 99%

Interpretation of In Vitro Concentration-Response Data for Risk Assessment and Regulatory Decision-making: Report from 2022 IWGT Quantitative Analysis Expert Working Group Meeting

Beal

Chen

Dearfield³

et al. 2023

Preprint

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Quantitative risk assessments of chemicals are routinely performed in rodents; however, there is growing recognition that non-animal approaches can be human-relevant alternatives. There is an urgent need to build confidence in non-animal alternatives given the international support to reduce the use of animals in toxicity testing where possible. In order for scientists and risk assessors to prepare for this paradigm shift in toxicity assessment, standardization and consensus on in vitro testing strategies and data interpretation will need to be established. To address this issue, an Expert Working Group (EWG) of the 8th International Workshop on Genotoxicity Testing (IWGT) evaluated the utility of quantitative in vitro genotoxicity concentration-response data for risk assessment. The EWG first evaluated available in vitro methodologies and then examined the variability and maximal response of in vitro tests to estimate biologically relevant values for the critical effect sizes considered adverse or unacceptable. Next, the EWG reviewed the approaches and computational models employed to provide human-relevant dose context to in vitro data. Lastly, the EWG evaluated risk assessment applications for which in vitro data are ready for use and applications where further work is required. The EWG concluded that in vitro genotoxicity concentration-response data can be interpreted in a risk assessment context. However, prior to routine use in regulatory settings, further research will be required to address the remaining uncertainties and limitations.

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Quantitative interpretation of ToxTracker dose–response data for potency comparisons and mode‐of‐action determination

Cited by 6 publications

References 43 publications

Systems for Genetic Assessment of the Impact of Environmental Factors

Systems for Genetic Assessment of the Impact of Environmental Factors

Interlaboratory validation of the ToxTracker assay: An in vitro reporter assay for mechanistic genotoxicity assessment

Interpretation of In Vitro Concentration-Response Data for Risk Assessment and Regulatory Decision-making: Report from 2022 IWGT Quantitative Analysis Expert Working Group Meeting

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