Abstract
PURPOSE: Hormonal contraceptives are linked to a higher prevalence of depressive symptoms. Given their popularity in Western countries, understanding the biochemical effects on neuronal cells is crucial to minimizing mental health risks. EXPERIMENTAL DESIGN: Neural progenitor cells were treated with ethinyl estradiol (EE) and levonorgestrel (LNG), two synthetic sex hormones commonly used in oral contraception, and S-23, a selective androgen receptor modulator developed as a potential synthetic sex hormone for male hormonal contraception. Label-based quantitative proteomics with the TMTpro 16plex tandem mass tags were used to assess protein expression changes between treated and untreated cells. RESULTS: Treatment of human neural progenitor cells with EE, LNG, EE + LNG, and S-23 led to distinct and overlapping proteomic changes, with enrichment in pathways related to inflammation, oxidative stress, transcriptional regulation, and cell death. Disease association analyses linked these changes to neurodegenerative and psychiatric conditions, including mechanisms relevant to depression. CONCLUSIONS AND CLINICAL RELEVANCE: These findings suggest that hormonal compounds used in contraception and performance enhancement may influence molecular pathways implicated in mental health, particularly depression. Although not directly translatable to clinical outcomes, the results support the need for further investigation into the neuropsychiatric effects of hormonal treatments. SUMMARY: This study addresses a pressing clinical need to better understand the potential mental health impacts of widely used hormonal contraceptives. While highly effective for pregnancy prevention, compounds such as ethinyl estradiol and levonorgestrel have repeatedly been associated with increased risk of depressive symptoms, highlighting the importance of investigating their molecular effects on neural systems. To explore this, we applied label-based quantitative proteomics in an undifferentiated human neural progenitor cell model treated with ethinyl estradiol, levonorgestrel, their combination, and the selective androgen receptor modulator S-23. The treatments induced distinct and overlapping changes in protein expression, with enrichment in pathways related to inflammation, oxidative stress, cell adhesion, chromatin dynamics, and programmed cell death-biological processes known to intersect with mechanisms implicated in depression. These findings offer insight into how synthetic hormones and hormone-like compounds may modulate neuronal biology, potentially contributing to adverse mental health outcomes. However, due to limitations of the in vitro model-such as the absence of systemic context, pharmacokinetics, and mature neuronal function-these results are primarily hypothesis-generating. They underscore the importance of further research to clarify the pathophysiological mechanisms linking hormonal treatments to affective disorders and to better assess the mental health risks of these compounds.