The NEMESIS project has made substantial progress toward its goal of elucidating how EDCs contribute to metabolic disruption, including insulin resistance and metabolic dysfunction-associated steatotic liver disease (MASLD) both in adults and in the developing individuals. Central to these efforts has been the successful establishment and deployment of a wide range of experimental models. These include advanced in vitro human systems such as 3D liver microtissues derived from primary hepatocytes as well as hepatic 3D co-cultures and pancreatic islet-like aggregates differentiated from human pluripotent stem cells.
Complementing these, in vivo work in mice and zebrafish have been initiated, enabling comparative analyses across species. Preliminary results suggest that the gut microbiome mediates part of the observed effects of MDCs, with changes in microbial composition and metabolite profiles observed in both model organisms following exposure. Sex-specific responses and developmental effects are being further investigated. The project has also advanced understanding of the interplay between the gut microbiome and MDC toxicity. In both mouse and zebrafish models, MDC exposure has been shown to significantly alter gut microbiota composition and the levels of microbiome-derived metabolites, which may in turn influence host metabolic health. Experiments using gnotobiotic zebrafish have demonstrated altered toxicity responses in the absence of a normal microbiota, highlighting the importance of microbiome-mediated mechanisms.
Mechanistic studies are well underway, employing transcriptomics and metabolomics to characterize the cellular and molecular impacts of MDC exposure. Dose-response analyses have been conducted for endpoints such as hepatic steatosis, mitochondrial function, and nuclear receptor activation. These mechanistic data are being integrated into a multi-omics framework that will enable computational modelling of metabolic disruption. A pilot study investigating 4β-hydroxycholesterol as a potential biomarker of EDC exposure is also ongoing, and additional multi-omics biomarker discovery studies are planned. In parallel, human cohort studies have been mobilized to support the project’s translational aims. EDC exposure data from several cohorts have been harmonized, and biological samples are being processed for exposure and metabolomics analyses. These activities will support the identification of early biological signals of MDC-induced metabolic perturbation, particularly in vulnerable populations.
Progress has been made in assessing the toxicity of individual MDCs as well as developing methodologies for mixture risk assessment. A comprehensive mapping of existing approaches for mixture assessment has been completed, and the initial steps toward evaluating the toxicity of realistic EDC mixtures relevant to EU populations have begun. These assessments are informed by toxicokinetic data and will incorporate AOPs and IATA frameworks, developed in collaboration with ENKORE working groups.
