A cell-based luminescence high-throughput screening to identify antivirals for EV-D68

Authors

• P Gao
• I Pavlinov
• M Xu
• X Huang
• Q Zhang
• M Shen
• CZ Chen
• W Zheng

Abstract

The NIH Antiviral Program for Pandemics (APP) was created in June 2021 as a response to the global pandemic of coronavirus disease 2019 (COVID-19). This program identified 11 viral families for developing safe and effective antivirals based on their pandemic potential. One of these families of interest is the Picornaviridae, a large family of small, positive-sense RNA viruses, that includes the genus Enterovirus. Enterovirus D68 (EV-D68), a non-polio enterovirus, is transmitted through the respiratory tract contrary to the fecal-oral route of poliovirus. It mainly infects children and causes flu-like symptoms and can lead to acute flaccid paralysis. In 2014, there was an EV-D68 outbreak in the USA. To date, no vaccine is available against enterovirus infection. In line with the APP goals, there is an urgency for safe antivirals against EV-D68, and we developed a cell viability-based, 1536-well assay for high throughput screening (HTS) to identify candidate antiviral compounds for enterovirus D68.
One of the EVD68-induced cytopathic effects in vitro is extensive cell death, therefore our HTS is based on a luminescence readout to measure intracellular ATP using CellTiter Glo (CTG) as a function of cell viability. Here, the optimized steps of the HTS will be described.
To validate our CTG-based EVD-68 assay, the NCATS Anti-infective library was screened. Several previously reported compounds were re-identified as antiviral in this screen. Our HTS is suitable to screen other NCATS libraries and will be used for drug repurposing other chemically diverse libraries and the development of computational modeling to predict anti-EV-D68 compounds.

Scientific Focus Area: Virology

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