Integrative analysis reveals multitarget associations of liquid crystal monomer in ovulatory dysfunction: insights from composite and subtype-specific Mendelian randomization.

Abstract

Objective: To investigate the potential pathogenic mechanisms of the liquid crystal monomer 4-ethyl-2-(4-(4-ethylbenzyl) phenoxy) benzonitrile (EBCN) in ovulatory dysfunction (composite phenotype including polycystic ovary syndrome [PCOS], anovulation, and primary ovarian insufficiency [POI]) through an integrative computational strategy that combines composite and subtype-specific Mendelian randomization.

Design: Integrative computational study combining network toxicology, Mendelian randomization, and molecular docking.

Subjects: For Mendelian randomization analysis, genetic data were derived from large cohorts of European ancestry, including plasma protein quantitative trait loci data and genome-wide association study summary statistics for a composite ovulatory dysfunction phenotype (FinnGen finngen_R12_E4_OVARDYS, 3,045 cases and 267,780 controls), which explicitly includes PCOS (E28.2), anovulatory infertility (N97.0), POI (E28.3), and other related diagnoses. Subtype-specific analyses were conducted for PCOS (finngen_R12_E4_PCOS, n_cases = 2,214) and POI (finngen_R12_E4_OVARFAIL, n_cases = 655).

Exposure: In silico exposure to EBCN for target prediction; genetically predicted plasma protein levels for causal inference.

Main outcome measures: Identification of overlapping targets between EBCN and ovulatory dysfunction; functional enrichment pathways; causal effects of prioritized proteins on ovulatory dysfunction risk (odds ratios [OR] with 95% confidence intervals); binding affinities between EBCN and core targets (kcal/mol).

Results: Network toxicology identified 150 overlapping targets between EBCN and ovulatory dysfunction. Enrichment analyses revealed significant associations with cancer-related pathways, PI3K-Akt and MAPK signaling cascades, and processes involving inflammation, metabolism, and apoptosis. Mendelian randomization demonstrated that genetically elevated levels of JUN (OR 1.265), CYP3A4 (OR 1.192), FGFR3 (OR 1.182), MET (OR 1.128), and POLR1C (OR 1.190) were significantly associated with increased risk of the composite ovulatory dysfunction phenotype, whereas higher PSAP levels were protective (OR 0.931). Subtype-specific analyses revealed convergent signals (e.g., CYP3A4 in PCOS) and divergent signals (e.g., TYMS as a risk factor in PCOS but protective in POI). Molecular docking confirmed stable binding interactions between EBCN and these targets, with strongest affinities for CYP3A4 (-8.9 kcal/mol) and JUN (-7.7 kcal/mol).

Conclusion: This integrative investigation identifies inflammatory regulators, xenobiotic-metabolizing enzymes, and cellular pathway proteins as potential mediators genetically associated with ovulatory dysfunction risk. However, whether EBCN directly affects these proteins and thereby contributes to ovulatory pathology remains to be experimentally validated. The findings provide prioritized targets and testable hypotheses for future experimental studies and environmental risk assessment.