Chemical safety assessment still predominantly relies on animal studies that focus on apical endpoints of toxicity. However, these traditional animal-centred approaches are being challenged owing to ethical and scientific concerns, including poor human predictivity and lack of mechanistic understanding (Vinken, 2024). To address these limitations, the early twenty-first century marked a paradigm shift in toxicology and moved away from animal-dependent assessment towards next-generation risk assessment. This exposure-led and hypothesis-driven approach strives to replace and/or reduce animal testing by applying new approach methodologies (NAMs) (Cronin et al., 2022; EFSA, 2018; Judson, 2013). NAMs refer to in vitro, in silico, and in chemico methods, as well as to approaches that refine in vivo methodologies. This supports the implementation of the 3R principle (Replacement, Reduction, and Refinement of animal experimentation) and facilitates risk assessment through humanrelevant techniques (Luijten et al., 2022; Schmeisser et al., 2023). NAMs move away from the ‘black box’ approach inherent to animal-based testing towards one where understanding the molecular mechanisms of toxicity becomes central (Kramer et al., 2019; Becker et al., 2017).