Mechanistic Insights into the Effects of Liquid Crystalline Monomers on Intestinal Stem Cell Differentiation Imbalance by Integrating Machine Learning and Adverse Outcome Pathway Framework Based on Organoids

The global annual output of liquid crystal monomers (LCMs) continues to increase, yet associated environmental and health risks remain poorly characterized. Assessing the toxic effects of the LCM mixture and individual components is critical for risk evaluation. Organoid models, characterized by highly biomimetic crypt-villus structures and cellular heterogeneity, serve as the preferred in vitro platforms for risk assessments. Our results reveal that exposure to the LCM mixture inhibits organoid expansion rates and induces abnormal crypt branching. Specifically, four machine learning models predicted that 4-cyano-4'-ethoxybiphenyl (EBCN) significantly contributes to intestinal toxicity. Exposure to EBCN alone reduces the proportion of viable cells, impairs intestinal barrier function, and disrupts stem cell homeostasis. Specifically, upregulated expression of the intestinal stem cell marker olfactomedin 4 (OLFM4) aligns with molecular signatures of inflammatory bowel disease patients in the Gene Expression Omnibus (GEO) data set. Additionally, the application of the Adverse Outcome Pathway framework identifies complement component C3 as the initial event, with downstream immunosuppression mediating EBCN-induced intestinal toxicity. In conclusion, this study highlights potential adverse effects of LCMs on gut physiology and emphasizes the urgent need for comprehensive health risk assessments for both the LCM mixture and individual LCM constituents.