Flavio Ballante

Chemical health risk assessment has failed to protect female fertility. Women are exposed to mixtures of synthetic chemicals linked to reduced ovarian function, lower embryo quality, and decreased fertility. Our previous FORMAS studies revealed new molecular targets in ovarian and endometrial cells exposed to chemicals in vitro, highlighting mechanisms related to metabolism, energy balance, and structural cellular biology, expanding the scope of reproductive toxicity beyond classical endocrine disruption. In HERTOX, we will develop these endpoints into high-throughput screening (HTS) assays using platforms originally designed for cancer drug testing. Human ovarian and endometrial cells will be cultured in 2D and 3D in 384-well formats, exposed to a customized library of up to 5 000 chemicals, and analyzed with multiplexed fluorescence imaging. Selected hits will undergo further testing for concentration-response properties, combinatorial toxicity, and validation through metabolomic and transcriptomic analyses. Structure-activity relationships will be analyzed, and the curated chemical library will be made available to researchers. Finally, selected exposures will be validated for their association with adverse outcomes in women using ovarian tissue cultures, a human stem cell-based embryo attachment model, and patient samples. HERTOX ambition is to deliver HTS systems predictive of reproductive toxicity in women.