The helminth parasite Fasciola hepatica modulates barrier integrity of human intestinal organoid-derived monolayers

Migration of the helminth parasite Fasciola hepatica across the intestine is a critical determinant of infection success, yet the molecular mechanisms governing this process remain poorly understood. Here, we apply human organoid-derived monolayers (ODMs) to investigate early interactions between F. hepatica newly excysted juveniles (FhNEJ) and the intestinal epithelium. ODMs were stimulated with live FhNEJ, and barrier function was assessed using measurements of transepithelial electrical resistance, fluorescein flux assays, and immunofluorescent detection of tight junction proteins. Parasite-induced changes in host protein expression were further examined by proteomics. Results showed that FhNEJ actively engaged with the ODM epithelium and induced a time- and dose-dependent loss of barrier integrity, accompanied by reduced expression and apical localisation of zonula occludens-1, a key tight junction component. Proteomic profiling of ODMs identified additional differentially regulated proteins involved in cell adhesion, immunity, and intercellular junction organisation. Together, these results provide novel insights into how FhNEJ compromise intestinal barrier permeability and establish ODMs as a robust and physiologically relevant platform for dissecting the molecular events that underlie the intestinal phase of F. hepatica infection.