Inducible POU2F3 expression in human intestinal organoids as a model of human tuft cells

Background & Aims: Tuft cells are relatively rare specialized chemosensory cells present in the intestinal epithelium. In mice, the intestinal tuft cell population expands in response to parasitic infection and stimulates the anti-parasitic immune response via cytokine signaling. However, elucidating the role of human intestinal tuft cells in homeostasis and disease remains has remained difficult due to their rarity in vivo and the lack of in vitro models. Methods: We genetically engineered human intestinal organoids (HIOs) to express POU2F3, a transcription factor that drives tuft cell specification, and created an HIO that is approximately 40% tuft cell composition. Results: Human tuft cells generated in vitro express known tuft cell-specific transcripts (AVIL, TRPM5, ALOX5, PLCG2) as well as exhibit characteristics in vivo tuft morphology. Organoids with tuft cells exhibit transcriptional signatures that suggest WNT signaling and the presence of non-motile cilium. The tuft cells were infectable with human rotavirus, which has recently been shown to be a new target of rotavirus infection in the intestine. Conclusions: The expression of POU2F3 in HIOs results in a novel model system for functional studies of human intestinal tuft cells.