Integrative network toxicology, multiomics, and machine learning elucidate zearalenone-induced developmental toxicity mechanisms in zebrafish (Danio rerio).

Zearalenone (ZEN) is a naturally occurring environmental contaminant that is produced by fungi. Developing animals are particularly sensitive to ZEN, while the mechanisms underlying ZEN-induced developmental toxicity remain incompletely understood. Understanding the developmental toxicity mechanisms of ZEN is crucial for safeguarding environmental and human health. Zebrafish serve as an excellent model organism for environmental toxicology research due to their short life cycle, sensitivity to environmental toxins, and transparent embryonic stage that allows direct observation of organ morphology during development. Therefore, this study employed zebrafish embryos as an experimental model, integrated network toxicology, transcriptomics, metabolomics, and machine learning for in-depth analysis. Phenotypic assessment confirmed that ZEN exposure leads to developmental arrest and organ abnormalities in zebrafish. Transcriptomics data revealed that ZEN primarily impairs zebrafish nervous development. Cross-analysis of the differentially expressed genes with network toxicology identified 54 key targets, primarily associated with cell fate determination and defense responses. Metabolomics and machine learning further identified key metabolites, such as pyruvic acid and nicotinamide adenine dinucleotide. Ultimately, this study further confirmed that energy metabolism and cell fate-related pathways are key pathways for ZEN-induced developmental toxicity by systematic multidimensional association analysis of genes, pathways, and metabolites. Overall, this study provides a comprehensive mechanistic framework for elucidating the developmental toxicity of ZEN and offers new insights for further identification of the targets and pathways involved in ZEN-induced developmental toxicity.