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Testing Strategies for Metabolite-Mediated Neurotoxicity.

Julian Suess1, Moritz Reinmoeller1, Viktoria Magel1

  • 1In Vitro Toxicology and Biomedicine, Dept Inaugurated by the Doerenkamp-Zbinden Foundation, University of Konstanz, 78457 Konstanz, Germany.

International Journal of Molecular Sciences
|September 13, 2025
PubMed
Summary

To improve next-generation risk assessment (NGRA), this study integrates metabolism into new approach methods (NAMs) for developmental neurotoxicity testing. This strategy addresses false negatives from assays lacking metabolic activity, enhancing human hazard prediction.

Keywords:
developmental neurotoxicitymicrosomesneurotoxicitytoxicity screeningxenobiotic metabolism

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Area of Science:

  • Toxicology
  • Developmental Neurotoxicity
  • In Vitro Assays

Background:

  • Metabolism-dependent toxicity challenges current non-animal new approach methods (NAMs) in risk assessment.
  • Existing NAMs often lack metabolic activity, potentially leading to false negative predictions for human health hazards.
  • Assessing metabolite-mediated toxicity is crucial for accurate developmental neurotoxicity (DNT) evaluations.

Purpose of the Study:

  • To develop and evaluate strategies for integrating metabolic activity into DNT NAMs.
  • To identify suitable positive controls for metabolite-related neurotoxicity.
  • To enhance the predictive capacity of NAMs for accurately assessing chemical risks.

Main Methods:

  • Utilized hepatic post-mitochondrial fractions (S9) to generate metabolite mixtures.
  • Applied metabolite mixtures to NAMs, including the MitoMet (UKN4b) and cMINC (UKN2) assays.
  • Assessed the toxic potency of metabolites compared to parent compounds on neuronal health.

Main Results:

  • Demonstrated that some metabolites exhibit higher neurotoxic potency than their parent compounds.
  • Successfully integrated a "metabolisation module" using S9 fractions into DNT NAMs.
  • Showcased the applicability and transferability of the integrated approach across different NAMs.

Conclusions:

  • The developed strategy effectively addresses metabolite-mediated toxicity in DNT NAMs.
  • Integrating metabolic capability into NAMs is essential for robust next-generation risk assessment (NGRA).
  • Further discussion is needed for broader implementation of this approach in regulatory science.