beta-D-N-4-Hydroxycytidine Is a Potent Anti-alphavirus Compound That Induces a High Level of Mutations in the Viral Genome

• Type of Material: Article
• Authors: Nadya Urakova, University of Alabama; Valeriya Kuznetsova, University of Alabama; David K. Crossman, University of Alabama; Arpine Sokratian, University of Alabama; David B. Guthrie, Emory University; Alexander A. Kolykhalov, Emory University; Mark A. Lockwood, Emory University; Michael George Natchus, Emory University; Michael R. Crowley, University of Alabama; George R. Painter, Emory University; Elena I. Frolova, University of Alabama; Illya Frolov, University of Alabama
• Language: English
• Date: 2018-02-01
• Publisher: American Society for Microbiology
• Publication Version: Final Published Version
• Copyright Statement: © 2018 American Society for Microbiology.
• Final Published Version (URL): http://dx.doi.org/10.1128/JVI.01965-17
• Title of Journal or Parent Work: Journal of Virology
• ISSN: 0022-538X
• Volume: 92
• Issue: 3
• Grant/Funding Information: This study was supported by Defense Threat Reduction Agency contract HDTRA1-15-C-0075 to G.R.P. and Public Health Service grants AI118867 to E.I.F. and AI095449 to I.F.
• Keywords: Virology; alphaviruses; ERROR CATASTROPHE; LETHAL MUTAGENESIS; VIRUS-RNA REPLICATION; N-hydroxycytidine; Venezuelan equine encephalitis virus; RNA-dependent RNA polymerase; antivirals; POLYMERASE; MOSQUITO CELLS; POLIOVIRUS; Science & Technology; Life Sciences & Biomedicine; VENEZUELAN EQUINE ENCEPHALITIS; drug-resistant mutant; FIDELITY; lethal mutagenesis; RIBAVIRIN RESISTANCE; ANTIVIRAL ACTIVITY
• Research Categories: Health Sciences, Pharmacology; Biology, Microbiology

Abstract

Venezuelan equine encephalitis virus (VEEV) is a representative member of the New World alphaviruses. It is transmitted by mosquito vectors and causes highly debilitating disease in humans, equids, and other vertebrate hosts. Despite a continuous public health threat, very few compounds with anti-VEEV activity in cell culture and in mouse models have been identified to date, and rapid development of virus resistance to some of them has been recorded. In this study, we investigated the possibility of using a modified nucleoside analog, β-D-N4-hydroxycytidine (NHC), as an anti-VEEV agent and defined the mechanism of its anti-VEEV activity. The results demonstrate that NHC is a very potent antiviral agent. It affects both the release of genome RNA-containing VEE virions and their infectivity. Both of these antiviral activities are determined by the NHC-induced accumulation of mutations in virus-specific RNAs. The antiviral effect is most prominent when NHC is applied early in the infectious process, during the amplification of negative- and positive-strand RNAs in infected cells. Most importantly, only a low-level resistance of VEEV to NHC can be developed, and it requires acquisition and cooperative function of more than one mutation in nsP4. These adaptive mutations are closely located in the same segment of nsP4. Our data suggest that NHC is more potent than ribavirin as an anti- VEEV agent and likely can be used to treat other alphavirus infections.