Modelling human development and disease in pluripotent stem-cell-derived gastric organoids

Abstract

Gastric diseases, including peptic ulcer disease and gastric cancer, affect 10% of the world's population and are largely due to chronic Helicobacter pylori infection1-3. Species differences in embryonic development and architecture of the adult stomach make animal models suboptimal for studyinghuman stomach organogenesis and pathogenesis4, and there is no experimentalmodel of normal human gastric mucosa. Here we report the de novo generation of threedimensional human gastric tissue in vitro through the directed differentiation of human pluripotent stemcells.Weshowthat temporal manipulation of the FGF,WNT,BMP, retinoic acid andEGFsignalling pathways and three-dimensional growth are sufficient to generate human gastric organoids (hGOs). Developing hGOs progressed through molecular and morphogenetic stages that were nearly identical to the developing antrum of the mouse stomach. Organoids formed primitive gastric gland-and pit-like domains, proliferative zones containing LGR5-expressing cells, surface and antral mucous cells, and a diversity of gastric endocrine cells.WeusedhGOcultures to identify novel signallingmechanisms that regulate early endoderm patterning and gastric endocrine cell differentiation upstreamof the transcription factorNEUROG3. UsinghGOs tomodel pathogenesis ofhuman disease, wefound thatH. pylori infection resulted in rapid association of the virulence factor CagA with the c-Met receptor, activation of signalling and induction of epithelial proliferation. Together, these studies describe a new and robust in vitro system for elucidating the mechanisms underlying human stomach development and disease.