New approach methodologies to assess reproductive risks of memory-enhancing dietary supplements - Proposal Abstract Dietary supplements, including memory-enhancing compounds such as vinpocetine, are widely available to consumers despite limited regulatory oversight on their reproductive safety. The U.S. Food and Drug Administration (FDA) has issued warnings regarding the potential teratogenicity of vinpocetine based on animal studies, yet human-relevant data remain scarce. This project aims to develop and apply pluripotent stem cell-based models as new approach methodologies (NAMs) to evaluate the teratogenicity of vinpocetine and related memory-enhancing supplements. These models, derived from both mouse and human stem cells, recapitulate key embryonic morphogenesis processes, enabling mechanistic investigations into teratogenic effects, as reported in our published studies. In Aim 1, we will characterize the molecular impact of vinpocetine on these morphogenesis models by performing whole-genome transcriptomic analyses. Bioinformatic analysis of differentially expressed genes will reveal molecular pathways that are disrupted by vinpocetine. In Aim 2, we will identify molecular targets of vinpocetine responsible for its teratogenicity. While the purported cognitive benefits of vinpocetine are linked to its inhibition of phosphodiesterase 1 and IκB kinase β, whether these targets mediate its teratogenicity remains unknown. To address this, we will perform loss-of-function studies for these two genes in our morphogenesis models and compare the resulting phenotypes to those treated with vinpocetine. In Aim 3, we will use our morphogenesis models to assess the potential teratogenicity of alternative memory-enhancing supplements. With vinpocetine identified by the FDA as a potential teratogen, consumers may turn to alternative supplements, many of which lack developmental and reproductive toxicity data. Using our morphogenesis models, we will evaluate the adverse effects of commonly marketed memory- enhancing supplements by analyzing concentration-effects relationships, which can be used to guide future safety assessment and regulatory actions for these supplements. This proposal combines innovative stem cell technology, transcriptomic analyses, and human-relevant endpoints to advance teratogenicity testing beyond traditional animal-based studies. Findings from this project will contribute to public health by informing regulatory considerations for dietary supplement safety. Additionally, this work will establish a versatile NAM platform to improve and accelerate the reproductive risk assessment of chemicals, offering cost-effective, time- efficient, and ethical alternative to conventional animal-based approaches.
