Denisa Bojkova, Philipp Reus, Leona Panosch, Marco Bechtel, Tamara Rothenburger, Joshua D. Kandler, Annika Eby Pfeiffer, Julian Uwe Gabriel Wagner, Mariana Shumliakivska, Stefanie Dimmeler, Ruth Maria Olmer, Ulrich Martin, Florian Vondran, Tuna Toptan, Florian Rothweiler, Richard Zehner, Holger Rabenau, Karen L. Osman, Steven T. Pullan, Miles Carroll, Richard Stack, Sandra Ciesek, Mark N. Wass, Martin Michaelis, Jindrich Cinatl
- Reliable, easy-to-handle phenotypic screening platforms are needed for the identification of anti-SARS-CoV-2 compounds. Here, we present caspase 3/7 activity as a read-out for monitoring the replication of SARS-CoV-2 isolates from different variants, including a remdesivir-resistant strain, and of other coronaviruses in a broad range of cell culture models, independently of cytopathogenic effect formation. Compared to other cell culture models, the Caco-2 subline Caco-2-F03 displayed superior performance, as it possesses a stable SARS-CoV-2 susceptible phenotype and does not produce false-positive hits due to drug-induced phospholipidosis. A proof-of-concept screen of 1796 kinase inhibitors identified known and novel antiviral drug candidates including inhibitors of PHGDH, CLK-1, and CSF1R. The activity of the PHGDH inhibitor NCT-503 was further increased in combination with the HK2 inhibitor 2-deoxy-D-glucose, which is in clinical development for COVID-19. In conclusion, caspase 3/7 activity detection in SARS-CoV-2-infected Caco-2F03 cells provides a simple phenotypic high-throughput screening platform for SARS-CoV-2 drug candidates that reduces false positive hits.
