Cell-based high-content approach for SARS-CoV-2 neutralization identifies unique monoclonal antibodies and PI3K pathway inhibitors

Carly Cabel¹Briana A Guzman¹Elaheh Alizadeh¹Shuaizhi Li¹Cameron Holberg¹Chonlarat Wichaidit¹Darren A Cusanovich¹Andrew L Paek¹Gregory R J Thatcher¹Koenraad Van Doorslaer¹Rachel S Nargi²Rachel E Sutton²Naveenchandra Suryadevara²James E. Crowe, Jr.²Robert H Carnahan²Samuel K Campos¹Curtis Andrew Thorne^1*

• ¹University of Arizona, Tucson, Arizona, United States
• ²Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Nashville, Tennessee, United States

The sudden rise of the SARS-CoV-2 virus and the delay in the development of effective therapeutics to mitigate it made evident a need for ways to screen for compounds that can block infection and prevent further pathogenesis and spread. Yet, identifying effective drugs efficacious against viral infection and replication with minimal toxicity for the patient can be difficult. Monoclonal antibodies were shown to be effective, yet as the SARS-CoV-2 mutated, these antibodies became ineffective. Small molecule antivirals were identified using pseudovirus constructs to recapitulate infection in non-human cells, such as Vero E6 cells. However, the impact was limited due to poor translation of these compounds in the clinical setting. This is partly due to the lack of similarity of screening platforms to the in vivo physiology of the patient and partly because drugs effective in vitro showed doselimiting toxicities. In this study, we performed two high-throughput screens in human lung adenocarcinoma cells with authentic SARS-CoV-2 virus to identify both monoclonal antibodies that neutralize the virus and clinically useful kinase inhibitors to block the virus and prioritize minimal host toxicity. Using high-content imaging combined with single-cell and multidimensional analysis, we identified antibodies and kinase inhibitors that reduce virus infection without affecting the host. Our screening technique uncovered novel antibodies and overlooked kinase inhibitors (i.e. PIK3i, mTORi, multiple RTKi) that could be effective against SARS-CoV-2 virus. Further characterization of these molecules will streamline the repurposing of compounds for the treatment of future pandemics and uncover novel mechanisms viruses use to hijack and infect host cells.