Abstract
Inhaled nebulized interferon (IFN)-α and IFN-β have been shown to be effective in the management of coronavirus disease 2019 (COVID-19). We aimed to construct a virus-free rapid detection system for high-throughput screening of IFN-like compounds that induce viral RNA degradation and suppress the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We prepared a SARS-CoV-2 subreplicon RNA expression vector which contained the SARS-CoV-2 5′-UTR, the partial sequence of ORF1a, luciferase, nucleocapsid, ORF10, and 3′-UTR under the control of the cytomegalovirus promoter. The expression vector was transfected into Calu-3 cells and treated with IFN-α and the IFNAR2 agonist CDM-3008 (RO8191) for 3 days. SARS-CoV-2 subreplicon RNA degradation was subsequently evaluated based on luciferase levels. IFN-α and CDM-3008 suppressed SARS-CoV-2 subreplicon RNA in a dose-dependent manner, with IC50 values of 193 IU/mL and 2.54 μM, respectively. HeLa cells stably expressing SARS-CoV-2 subreplicon RNA were prepared and treated with the IFN-α and pan-JAK inhibitor Pyridone 6 or siRNA-targeting ISG20. IFN-α activity was canceled with Pyridone 6. The knockdown of ISG20 partially canceled IFN-α activity. Collectively, we constructed a virus-free rapid detection system to measure SARS-CoV-2 RNA suppression. Our data suggest that the SARS-CoV-2 subreplicon RNA was degraded by IFN-α-induced ISG20 exonuclease activity.
Highlights
We identified that CDM-3008 suppresses cccDNA in hepatitis B virus (HBV)-infected primary cultured human hepatocytes; we named CDM after the cccDNA modulator [5]
We constructed a system for measuring the SARS-CoV-2 genomic RNA degradation activity of IFN-α within 1 day
SARS-CoV-2 genomic partly canceled by ISG20 siRNA. These results suggest that SARS-CoV-2 genomic RNA isis degraded by by ISG20
Summary
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Coronavirus disease (COVID-19) is a worldwide pandemic, and the development of therapeutic agents that target severe acute respiratory syndrome coronavirus 2. The pandemic has been suppressed by the development of vaccines, it persists, and there are concerns regarding the emergence of mutant strains that weaken vaccine efficacy. New coronaviruses, such as Middle East respiratory syndrome coronavirus (MERS-CoV), severe acute respiratory syndrome coronavirus (SARS-CoV), and SARS-CoV-2, have emerged at a frequency of approximately once per decade [1,2]. The development of therapeutic agents that are effective against any coronavirus is important. We focused on small chemical compounds that enhance the degradation of SARS-CoV-2 genomic RNA
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