Intestinal Enteroendocrine Lineage Cells Possess Homeostatic and Injury-Inducible Stem Cell Activity

Summary Several cell populations have been reported to possess intestinal stem cell (ISC) activity during homeostasis and injury-induced regeneration. Here, we explored inter-relationships between putative mouse ISC populations by comparative RNA-sequencing (RNA-seq). The transcriptomes of multiple cycling ISC populations closely resembled Lgr5 + ISCs, the most well-defined ISC pool, but Bmi1-GFP + cells were distinct and enriched for enteroendocrine (EE) markers, including Prox1 . Prox1-GFP + cells exhibited sustained clonogenic growth in vitro , and lineage-tracing of Prox1 + cells revealed long-lived clones during homeostasis and after radiation-induced injury in vivo . Single-cell mRNA-seq revealed two subsets of Prox1-GFP + cells, one of which resembled mature EE cells while the other displayed low-level EE gene expression but co-expressed tuft cell markers, Lgr5 and Ascl2 , reminiscent of label-retaining secretory progenitors. Our data suggest that the EE lineage, including mature EE cells, comprises a reservoir of homeostatic and injury-inducible ISCs, extending our understanding of cellular plasticity and stemness.