A novel organoid model of damage and repair identifies HNF4α as a critical regulator of intestinal epithelial regeneration

ABSTRACT Background & Aims Recent evidence suggests that the intact intestinal epithelial barrier protects our body from a range of immune-mediated diseases. The epithelial layer has an impressive ability to reconstitute and repair upon damage and this process of repair is increasingly seen as a therapeutic target. In-vitro models to study this process in primary intestinal cells are lacking. Methods Therefore we established and characterized an in-vitro model of intestinal damage and repair by applying γ-radiation on small-intestinal organoids. We then used this model to identify novel regulators of intestinal regeneration. Results We identified Hepatocyte Nuclear Factor 4 alpha (HNF4α) as a pivotal upstream regulator of the intestinal regenerative response. Organoids lacking Hnf4a were not able to propagate in vitro. Importantly, intestinal Hnf4a knock-out mice showed impaired regeneration after whole-body irradiation, confirming intestinal organoids as a valuable alternative to in-vivo studies. Conclusions In conclusion, we established and validated an in-vitro damage–repair model and identified HNF4α as a crucial regulator of intestinal regeneration.