During the first 18 months, HYPIEND has made substantial progress in advancing scientific knowledge and technical capacity to assess and mitigate the effects of EDC mixtures on the hypothalamic-pituitary (HP) axis. A robust data-driven framework for mixture selection was established, leading to the identification of thirteen individual EDCs and two candidate EDC mixtures based on co-exposure patterns and QSAR modelling. These two predefined mixtures, along with potential combinations derived from the 13 selected EDCs, are now set to be validated in vitro; however, the results of this stage have been delayed from Month 18 to Month 21. To support this phase, human cell-based assays are being used to characterize cytotoxic, estrogenic, and androgenic effects. In vivo zebrafish models have also been implemented to assess phenotypic disruptions, and one new transgenic zebrafish line is being developed. Organotypic models of the hypothalamus and pituitary were successfully developed, enabling early mechanistic insights into HP disruption. In parallel, in silico models predicting placental and blood-brain barrier permeability were implemented, complementing experimental barrier models. Preparatory work has been implemented to allow rodent experimentation, including approval from Ethical Committes, protocol standardization and establishment of transgenic lines. Foundational work on DNA methylation profiling and extracellular vesicle characterization has begun to support the development of translational biomarkers. Preparatory work was also completed for large-scale interventional studies, including finalization of study protocols and recruitment strategies, approval from Ethical Commitees (including the preparation of the corresponding legal documentation), development of the behavioural multi-component intervention and the booklets for the control groups, other preparatory activities (harmonization of lab material, samples codification, Redcap platform...) initial deployment of the perinatal intervention and recruitment of schools for the prepubertal intervention. Altogether, these activities position HYPIEND to achieve its mid- and long-term scientific goals through a fully integrated translational research pipeline.
