iScience (Feb 2023)
Identification of novel antiviral drug candidates using an optimized SARS-CoV-2 phenotypic screening platform
- Denisa Bojkova,
- Philipp Reus,
- Leona Panosch,
- Marco Bechtel,
- Tamara Rothenburger,
- Joshua D. Kandler,
- Annika Pfeiffer,
- Julian U.G. Wagner,
- Mariana Shumliakivska,
- Stefanie Dimmeler,
- Ruth Olmer,
- Ulrich Martin,
- Florian W.R. Vondran,
- Tuna Toptan,
- Florian Rothweiler,
- Richard Zehner,
- Holger F. Rabenau,
- Karen L. Osman,
- Steven T. Pullan,
- Miles W. Carroll,
- Richard Stack,
- Sandra Ciesek,
- Mark N. Wass,
- Martin Michaelis,
- Jindrich Cinatl, Jr.
Affiliations
- Denisa Bojkova
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany
- Philipp Reus
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany; Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany
- Leona Panosch
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany
- Marco Bechtel
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany
- Tamara Rothenburger
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany
- Joshua D. Kandler
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany
- Annika Pfeiffer
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany
- Julian U.G. Wagner
- Institute for Cardiovascular Regeneration, Centre of Molecular Medicine, Goethe University, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany; German Center for Cardiovascular Research (DZHK), partner site Rhine-Main, Goethe University, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany; Cardiopulmonary Institute (CPI), Goethe University, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany
- Mariana Shumliakivska
- Institute for Cardiovascular Regeneration, Centre of Molecular Medicine, Goethe University, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany
- Stefanie Dimmeler
- Institute for Cardiovascular Regeneration, Centre of Molecular Medicine, Goethe University, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany; German Center for Cardiovascular Research (DZHK), partner site Rhine-Main, Goethe University, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany; Cardiopulmonary Institute (CPI), Goethe University, Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany
- Ruth Olmer
- Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery (HTTG), REBIRTH-Research Center for Translational Regenerative Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center for Lung Research (DZL), Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Germany
- Ulrich Martin
- Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Department of Cardiothoracic, Transplantation and Vascular Surgery (HTTG), REBIRTH-Research Center for Translational Regenerative Medicine, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center for Lung Research (DZL), Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Germany
- Florian W.R. Vondran
- Department for General, Visceral and Transplant Surgery, Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Germany; German Center for Infection Research, DZIF, Braunschweig, Inhoffenstraße 7, 38124 Braunschweig, Germany
- Tuna Toptan
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany
- Florian Rothweiler
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany; Dr Petra Joh Research Institute, Komturstraße 3A, 60596 Frankfurt am Main, Germany
- Richard Zehner
- Institute for Forensic Medicine, University Hospital, Goethe-University, Kennedyallee 104, 60596 Frankfurt am Main, Germany
- Holger F. Rabenau
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany
- Karen L. Osman
- UK Health Safety Agency, National Infection Service, Porton Down, Salisbury, Wiltshire SP4 0JG, UK
- Steven T. Pullan
- UK Health Safety Agency, National Infection Service, Porton Down, Salisbury, Wiltshire SP4 0JG, UK
- Miles W. Carroll
- Wellcome Trust Centre for Human Genetics, Nuffield Department of Medicine, Oxford University, Roosevelt Drive, Headington, Oxford OX3 7BN, UK; NIHR Health Protection Unit in Emerging and Zoonotic Infections, Department of Clinical Infection, Microbiology and Immunology, University of Liverpool, Liverpool L69 7BE, UK
- Richard Stack
- School of Biosciences, University of Kent, Canterbury CT2 7NJ, UK
- Sandra Ciesek
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany; Fraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Theodor-Stern-Kai 7, 60596 Frankfurt am Main, Germany; German Center for Infection Research, DZIF, Braunschweig, Inhoffenstraße 7, 38124 Braunschweig, Germany
- Mark N. Wass
- School of Biosciences, University of Kent, Canterbury CT2 7NJ, UK
- Martin Michaelis
- School of Biosciences, University of Kent, Canterbury CT2 7NJ, UK; Corresponding author
- Jindrich Cinatl, Jr.
- Institute of Medical Virology, University Hospital, Goethe-University, Paul Ehrlich-Straße 40, 60596 Frankfurt am Main, Germany; Dr Petra Joh Research Institute, Komturstraße 3A, 60596 Frankfurt am Main, Germany; Corresponding author
- Journal volume & issue
-
Vol. 26,
no. 2
p. 105944
Abstract
Summary: 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 readout for monitoring the replication of SARS-CoV-2 isolates from different variants, including a remdesivir-resistant strain, and of other coronaviruses in numerous cell culture models, independently of cytopathogenic effect formation. Compared to other models, the Caco-2 subline Caco-2-F03 displayed superior performance. It possesses a stable SARS-CoV-2 susceptibility phenotype and does not produce false-positive hits due to drug-induced phospholipidosis. A proof-of-concept screen of 1,796 kinase inhibitors identified known and novel antiviral drug candidates including inhibitors of phosphoglycerate dehydrogenase (PHGDH), CDC like kinase 1 (CLK-1), and colony stimulating factor 1 receptor (CSF1R). The activity of the PHGDH inhibitor NCT-503 was further increased in combination with the hexokinase II (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-2-F03 cells provides a simple phenotypic high-throughput screening platform for SARS-CoV-2 drug candidates that reduces false-positive hits.
