Background: Chronic gastrointestinal disorders, such as inflammatory bowel disease (IBD), demand advanced research models to uncover their origins and potential treatments. To enhance our understanding and explore therapeutic options, it’s crucial to create translational models that faithfully replicate key aspects of IBD. Three-dimensional (3D) canine intestinal organoids, composed of diverse differentiated intestinal epithelial cells, present a promising avenue for such research.
Objectives: To investigate the effect of proinflammatory cytokines on canine intestinal epithelial using normal canine organoids.Animals and
Methods: Canine organoids were developed using colonic tissues biopsied from three healthy client-owned dogs. Subsequently, we initiated cytokine treatment (30 ng/mL) and analyzed the immunofluorescent staining of tight junction protein ZO-1 at 48 hours after exposure and leucine-rich repeat-containing G-protein (Lgr5) gene expression at 24 and 48 hours after exposure, comparing it to non-treated control organoids.
Results: We observed a significant decrease of fluorescent intensity of ZO-1 after three cytokines (p<0.001). And we also observe a significant decrease in Lgr5 mRNA quantitative PCR expression at 24 and 48 hours after treatment with IFN-γ (p<0.05 and p</em><0.001). Conversely, there were no significant differences observed between the TNF-α-treated, IL-1β-treated and the control group.Conclusions and clinical importance: This finding suggests that TNF-α, IFN-γ and IL-1β inhibit the tight junction protein and IFN-γ inhibits proliferative potential of canine intestinal organoids, indicating the potential therapeutic target of canine IBD. Our ability to observe these epithelial responses using canine organoids underscores the potential utility of these 3D organoids as an in vitro model for canine IBD.
