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 H. pylori infection1–3. Species differences in embryonic development and architecture of the adult stomach make animal models suboptimal for studying human stomach organogenesis and pathogenesis4, and there is no experimental model of normal human gastric mucosa. Here we report the de novo generation of three-dimensional human gastric tissue in vitro through the directed differentiation of human pluripotent stem cells (hPSCs). We identified that temporal manipulation of the FGF, WNT, BMP, retinoic acid and EGF signaling 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. We used hGO cultures to identify novel signaling mechanisms that regulate early endoderm patterning and gastric endocrine cell differentiation upstream of the transcription factor NEUROG3. Using hGOs to model pathogenesis of human disease, we found that H. pylori infection resulted in rapid association of the virulence factor CagA with the c-Met receptor, activation of signaling and induction of epithelial proliferation. Together, these studies describe a novel and robust in vitro system for elucidating the mechanisms underlying human stomach development and disease.

Modeling infectious diseases and host-microbe interactions in gastrointestinal organoids

“…In cancer cell line models, CagA binds to the host c-MET receptor and promotes a motogenic response (Churin et al, 2003). In gastric organoids, CagA also binds to c-Met and likely as a response, host cell proliferation is induced in a CagA-dependent manner (Bertaux-Skeirik et al, 2015;McCracken et al, 2014). Together, these studies establish organoids as a model for H. pylori infection by demonstrating that organoids reproduce important hallmarks of the infection.…”

“…As an alternative model lying between 2D cell lines and animals, several studies have now established H. pylori infection in gastric organoids. To enable interaction with the apical side of the cell, which is also the natural site of infection, H. pylori are microinjected into the lumen of the organoids McCracken et al, 2014). Injected bacteria can adhere to the epithelium and target apical cell-cell junctions Wroblewski et al, 2010).…”

Translational potential of human brain organoids

“…This landmark paper established a novel experimental paradigm for generating tissue organoids. Subsequent studies worldwide confirmed the generalizability of such approach to human stem cells, and produced organoids of a variety of organ types including colon,11 intestine,12 prostate,13, 14 fallopian tube,15 stomach,16 liver,17, 18 kidney,19 lung,20 and brain 8…”

The boom in mini stomachs, brains, breasts, kidneys and more

“…That excitement is shared by developmental biologist James Wells, who last year reported that he and his team had created an organoid that resembled part of a human stomach 7 . Wells started with a different raw material to Clevers, whose organoids arise from adult stem cells that can generate only a limited number of cell types.…”