Optimization of a Micro-scale Air-Liquid-Interface Model of Human Proximal Airway Epithelium for Moderate Throughput Drug Screening for SARS-CoV-2

Respiratory viruses (e.g. influenza, RSV, SARS, MERS etc.) attack the proximal airway (upper respiratory tract) and cause a wide spectrum of diseases for which we have limited therapies. To date, a few primary human stem cell-based models of the proximal airway have been reported for drug discovery but scaling them up to a high throughput platform remains a significant challenge. Here we present a microscale, primary human stem cell-based proximal airway model of SARS-CoV-2 infection, which is amenable to moderate-to-high throughput drug screening. The model recapitulates the heterogeneity of infection seen among different patients and with different SARS-CoV-2 variants. Using this model, we screened 2100 compounds from targeted drug libraries. We characterized a new small molecule with antiviral properties that is effective against both Wuhan and Omicron variants Overall design: Primary human airway basal stem cells were cultured in 96-well ALI plates to develop differentiated mucociliary epthelium. Cells were treated or not with Remdesivir and compound MKGaA#49, then exposed or not to Wuhan variant of SARS-CoV-2 for 72 hours. Cells were then harvested for total RNA sequencing