Modeling lamotrigine-induced reprotoxicity in porcine endometrial organoids: Integrated multi-platform profiling

Lamotrigine, a newer generation anti-epileptic drug aimed at addressing reproductive complications, requires thorough evaluation of its effects on the endometrium. Using the three-dimensional endometrial organoid (EO) model provides a distinct advantage in modeling lamotrigine-induced toxicity, offering a more relevant physiological system. In this study, a porcine EO model was used and treated with lamotrigine to mimic and analyze drug-induced toxicity. Porcine uteri were processed and digested with collagenase, then combined with Matrigel and incubated with 5 % COenvironment, at 38°C. During passaging, cells were dissociated, treated with trypsin-EDTA, and subcultured, with the medium renewed every 2-3 days. Different analytical methods were employed to evaluate lamotrigine's impact on the endometrial organoids, covering aspects such as cell viability, morphology, replication, steroidogenesis, and metabolic changes. The results showed significant alterations in cell morphology with a decrease in number and size. Metabolite analysis revealed metabolic shifts in some amino acids, glucose and galactose, ranging from approximately 1.5 to 5 times, (p < 0.05), when compared to the control groups. Molecular assays indicated increased oxidative stress, activation of apoptotic pathway, and disrupted steroidogenesis, revealing lamotrigine as an active endocrine disruptor. Moreover, lamotrigine induced changes in specific miRNAs that regulate implantation, and epithelial-mesenchymal transition pathways. In conclusion, our study highlights the potential diverse impact of lamotrigine on the endometrial microenvironment, emphasizing the need for further investigations into its implications on reproductive health and embryo implantation.2