Chan, Ryan, Chuenchob, Eve, Moquin, Stephanie, Manjunatha, Ujjini, Jarrousse Coscoy, Nadine, Menachery, Vineet, Xie, Xuping, Flannery, Erika, Eastman, Rich and Shema Mugisha, Christian (2024) Rapid-response RNA-FISH assay platform for coronavirus antiviral high-throughput screening. SLAS Discovery, 29 (8). pp. 297-303. ISSN 24725552

Abstract

Over the past 25 years, the global community has faced challenges posed by three distinct outbreaks of coronaviruses. The first of these outbreaks was the severe acute respiratory syndrome coronavirus (SARS-CoV) epidemic, which occurred from 2002 to 2004 and resulted in over 700 deaths. Following this, the Middle East respiratory syndrome coronavirus (MERS-CoV) epidemic emerged in 2012, causing over 2600 infections with a case-fatality rate of 36%. More recently, the world has become severely impacted by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) of the COVID-19 pandemic, responsible for causing nearly 7 million deaths worldwide. It is crucial to note that the threat of further coronavirus outbreaks continues to be a significant concern, as evidenced by the recent identification of the novel canine CoV (CCoV-HuPn-2018) in several patients with pneumonia in Malaysia. The threat of the ever-evolving nature of viral infections as well as the lingering health and socioeconomic effects of the recent SARS-CoV-2 pandemic emphasize the urgent need for advanced antiviral drug screening tools to strengthen preparedness and preventive measures against future outbreaks. Here, we present the development and validation of a novel RNA-fluorescence in situ hybridization (FISH) assay as a high-throughput rapid response platform for antiviral drug discovery. The flexibility of RNA-FISH probe sets allows for design of viral genome specific probes, enabling in vitro assay development to test for inhibition of viral replication by either biologic or small molecule inhibitors quickly. Screening of 170 antiviral compounds in concentration-response demonstrates a strong R2 correlation between the RNA-FISH assay and a gold-standard immunofluorescence assay for both human CoVs OC43 and 229E. Additionally, we successfully applied this methodology in the context of CCoV 1-71, proving rapid development and deployment, thus, opening new avenues for the evaluation of antiviral drugs to future emerging threats.

Item Type:	Article
Date Deposited:	16 Nov 2024 00:45
Last Modified:	16 Nov 2024 00:45
URI:	https://oak.novartis.com/id/eprint/54440