Significance: Infection by the bacterium Helicobacter pylori is the main risk factor for stomach cancer. Strains of H. pylori that produce the virulence factor CagA substantially increase stomach cancer risk compared with strains without CagA. CagA disrupts cell polarity—the specialized asymmetric organization within cells—but the mechanism it uses and its influence on colonization are not fully understood. Using a three-dimensional culture of stomach cells (human gastric organoids) and cell lines, we find that the tumor suppressor ASPP2 is crucial in disruption of cell polarity by CagA during H. pylori infection. Interfering with the CagA–ASPP2 interaction, using small molecules or a specific peptide, blocks loss of cell polarity and reduces bacterial colonization in organoids.

Enteroviruses are a leading cause of upper respiratory tract, gastrointestinal, and neurological infections. Management of enterovirus-related diseases has been hindered by the lack of specific antiviral treatment. The pre-clinical and clinical development of such antivirals has been challenging, calling for novel model systems and strategies to identify suitable pre-clinical candidates. Organoids represent a new and outstanding opportunity to test antiviral agents in a more physiologically relevant system. However, dedicated studies addressing the validation and direct comparison of organoids versus commonly used cell lines are lacking. Here, we described the use of human small intestinal organoids (HIOs) as a model to study antiviral treatment against human enterovirus 71 (EV-A71) infection and compared this model to EV-A71-infected RD cells. We used reference antiviral compounds such as enviroxime, rupintrivir, and 2′-C-methylcytidine (2′CMC) to assess their effects on cell viability, virus-induced cytopathic effect, and viral RNA yield in EV-A71-infected HIOs and cell line. The results indicated a difference in the activity of the tested compounds between the two models, with HIOs being more sensitive to infection and drug treatment. In conclusion, the outcome reveals the value added by using the organoid model in virus and antiviral studies.