Human respiratory organoids sustained reproducible propagation of previously uncultivable human rhinovirus C

The lack of a robust system to reproducibly propagate HRV-C substantially hampered our understanding of the common respiratory virus. We sought to develop an organoid-based system to reproducibly propagate HRV-C and characterize virus-host interaction using the respiratory organoids established by our team. We demonstrated that airway organoids sustained serial virus passage with the aid of CYT-387-mediated immunosuppression; nasal organoids, an organoid model more closely simulating the human upper airway, achieved this without any intervention. Nasal organoids were more susceptible to HRV-C than airway organoids. Intriguingly, we observed a more intensive innate immune response in airway organoids than nasal organoids upon HRV-C infection, which was reproduced in a Poly (I:C) stimulation assay. Treatment with an anti-CDHR3 and two antivirals significantly reduced HRV-C viral growth in nasal organoids. An organoid-based immunofluorescence assay was established to titrate HRV-C infectious particles. Collectively, we developed an organoid-based system to reproducibly propagate the previously uncultivable HRV-C, which enabled an in-depth elucidation of HRV-C infection and innate immunity unprecedentedly. The organoid-based HRV-C infection model can be extended for developing antiviral strategies. More importantly, our study has paved a new avenue for propagating and studying other uncultivable human and animal viruses.