Mature tuft cell phenotypes are sequentially expressed along the intestinal crypt-villus axis following cytokine-induced tuft cell hyperplasia

Intestinal tuft cells are epithelial sentinels that trigger host defense upon detection of parasite-derived compounds. While representing interesting targets for immunomodulatory therapies in inflammation-driven intestinal diseases, their detailed functioning is poorly understood. Although two distinct intestinal tuft cell types have been described, we reveal common intermediary transcriptomes among tuft cells in mouse and human. Tuft cell-specific reporter knock-ins in organoids show that the two tuft types are sequentially expressed transcriptomic states that represent different maturation stages. Moreover, cytokines interleukin-4 and interleukin-13 only induce lineage specification to Nrep + tuft-1 cells, while BMP and cholinergic signalling advance differentiation towards immune-related ChAT + tuft-2 phenotypes. Functionally, both tuft cell states have chemosensory capacity and respond to stimuli like succinate, but reaction probability increases during tuft cell maturation. Our tuft type-specific reporters and optimized differentiation strategy in organoids provide an experimental platform to study the functioning of tuft cells and their unique chemosensory properties.