Antiviral Agents Against African Swine Fever Virus T ⁎ Erik Arabyan, Armen Kotsynyan, Astghik Hakobyan, Hovakim Zakaryan
Total Page：16
File Type：pdf, Size：1020Kb
Virus Research 270 (2019) 197669 Contents lists available at ScienceDirect Virus Research journal homepage: www.elsevier.com/locate/virusres Review Antiviral agents against African swine fever virus T ⁎ Erik Arabyan, Armen Kotsynyan, Astghik Hakobyan, Hovakim Zakaryan Group of Antiviral Defense Mechanisms, Institute of Molecular Biology of NAS, Yerevan, Armenia ARTICLE INFO ABSTRACT Keywords: African swine fever virus (ASFV) is a significant transboundary virus that continues to spread outside Africa in African swine fever virus Europe and most recently to China, Vietnam and Cambodia. Pigs infected with highly virulent ASFV develop a Antiviral hemorrhagic fever like illness with high lethality reaching up to 100%. There are no vaccines or antiviral drugs Flavonoid available for the prevention or treatment of ASFV infections. We here review molecules that have been reported Nucleoside to inhibit ASFV replication, either as direct-acting antivirals or host-targeting drugs as well as those that act via a Pig yet unknown mechanism. Prospects for future antiviral research against ASFV are also discussed. 1. Introduction as HIV and hepatitis B virus infections. Nevertheless, there is still no antiviral drug for more than 200 viruses affecting human populations African swine fever (ASF) is one of the most important swine dis- worldwide (De Clercq and Li, 2016). Moreover, control of some animal eases due to its significant socioeconomic consequences for affected viruses like ASFV by means of an antiviral therapy appears to be an countries. It was first observed in Kenya in the early 20th century fol- attractive approach due to the lack of other control measures. There- lowing the introduction into the country of European domestic swine. fore, the development of antiviral agents for mass application in ve- Montgomery (1921) described the disease as an entity distinct from terinary is as much important as the development of antiviral drugs classical swine fever. Shortly after Kenya, reports of ASF in South Africa against human viruses. and Angola followed (Steyn, 1928; Gago da Câmara, 1933). Since then, In this review, a general introduction to the replication cycle of ASF were reported in many African countries, particularly in the sub- African swine fever virus (ASFV) is presented, followed by an overview Saharan region, where it still remains within wild and domestic pig of antiviral agents with identified and unknown targets and currently populations. It occurred in Europe (Portugal and Spain) in 1957 and available methods for antiviral drug testing. Future directions for de- 1960, and from there it spread to other European countries. However, veloping antiviral agents against ASFV is also discussed. ASF was eradicated from Europe in the mid-1990s, except for Sardinia, where it has been endemic since 1978 (Gallardo et al., 2015). In 2007, a 2. Replication cycle of ASFV second reintroduction of ASF to the European Continent occurred in Georgia from where it quickly spread to neighboring countries. ASF has ASFV is a large double-stranded DNA virus that belongs to the fa- been reported in Russia, Ukraine, Poland, Latvia, Lithuania, Estonia, mily Asfarviridae. In infected pigs, ASFV replicates in cells of the Moldova, Czech Republic, Romania, Hungary and most recently in mononuclear phagocyte system, mostly in macrophages and mono- China, Vietnam and Cambodia. As a result of ASF outbreaks in the cytes. The ASFV replication cycle starts with viral attachment and entry Russian and Eastern Europe, more than 800,000 pigs were slaughtered into the host cell. Although the identity of cellular receptors involved in from 2014 to 2017 (Sánchez-Cordón et al., 2018). The value of exports ASFV entry remains unknown, the receptor-dependent mechanism of of pork in Eastern Europe was reduced more than EUR 950 million viral entry, such as clathrin-mediated endocytosis has been reported (Cisek et al., 2016; Sánchez-Cordón et al., 2018; Chenais et al., 2019). (Galindo et al., 2015). There is also evidence that ASFV utilizes an al- Further spread and huge economic losses seems highly possible due to ternative mechanism, macropinocytosis, when it enters into Vero cells the lack of effective vaccines and control measures. (Sánchez et al., 2012). Within the endosome, ASFV undergoes con- Since the first antiviral drug, idoxuridine, approved in 1963, great formational changes due to the low pH environment. Viral uncoating, achievements have been made in the field of antiviral drug discovery. the loss of outer membrane and protein capsid, occurs in multivesicular Today more than 90 antiviral drugs categorized into 13 functional late endosomes, followed by the fusion of inner membrane with en- groups are available for the treatment of human infectious diseases such dosomes and the release of nude core to the cytosol (Hernáez et al., ⁎ Corresponding author at: Institute of Molecular Biology of NAS, Hasratyan 7, 0014, Yerevan, Armenia. E-mail address: [email protected] (H. Zakaryan). https://doi.org/10.1016/j.virusres.2019.197669 Received 14 June 2019; Received in revised form 15 July 2019; Accepted 16 July 2019 Available online 17 July 2019 0168-1702/ © 2019 Elsevier B.V. All rights reserved. E. Arabyan, et al. Virus Research 270 (2019) 197669 2016). Incoming ASFV particles are then localized near the nucleus, is an excellent model for antiviral drug research and is widely used by where they form the viral factory. On this site, newly synthesized viral different laboratories. DNA and proteins are accumulated. According to their accumulation kinetics, four classes of ASFV transcripts are known. Transcription of 3.2. Methods to evaluate in vitro antiviral activity immediately early and early genes starts immediately after infection and continues until the initiation of viral DNA synthesis. The transcription Two assays, plaque and cell viability/cytopathic effect (CPE) re- of two remaining classes, intermediate and late genes, occurs after DNA duction assays, are usually employed for the initial identification of replication is initiated (Rodríguez and Salas, 2013). Finally, mature molecules with antiviral activity against ASFV. Virus-induced CPE assay ASFV particles are transported from the viral factory to the cell surface can be observed microscopically or quantified by colorimetric mea- via a microtubule-mediated mechanism, which depends on the capsid surements using MTT, crystal violet stain or neutral red dye uptake protein pE120R and the motor protein conventional kinesin (Andrés methods. The advantage of colorimetric measurements is the possibility et al., 2001; Jouvenet et al., 2004). They exit the host cell by budding at of both discovering new antiviral agents and evaluating their cytotoxic the membrane, allowing extracellular virions to acquire an additional effects using the same method. lipid membrane. At very late times of infection, ASFV causes cell lysis, Recently, a new method for real-time monitoring of the CPE of ASFV which may represent an alternative mechanism of viral egress (Breese has been reported (Burmakina et al., 2018). This method is based on and DeBoer, 1966). microelectronic biosensor technology (xCELLigence® system developed Based on their anti-ASFV mechanisms, potential antiviral agents can by ACEA Inc.) allowing real time, label-free and non-invasive analysis be divided into two groups: (1) inhibitors that directly act on ASFV by of cellular changes including cell viability, proliferation and CPE. The targeting the viral proteins (direct-acting antivirals), or (2) inhibitors application of this real-time novel system as a straightforward and ef- that target cellular factors involved in ASFV replication (host-targeting ficient assay to measure the CPE has been demonstrated for several antivirals). Since anti-ASFV drug discovery largely depends on the DNA and RNA viruses (Fang et al., 2011; Marlina et al., 2015; Piret ® fundamental knowledge of virus replication, further research on the et al., 2016). Burmakina et al. (2018) showed that the xCELLigence role of viral enzymes and host factors remains needed. In the following system is able to detect measurable changes even though cells are in- section, we will provide a brief review of laboratory methods to study fected with ASFV strains that do not develop clear CPE, indicating that antiviral agents against ASFV. this method can be utilized in those studies where CPE-based assays are not applicable. 3. Methods for evaluation of antiviral compounds 4. Inhibitors with identified targets 3.1. Cells and viruses 4.1. Nucleoside analogues For many years, ASFV studies were only performed on the natural target cells, porcine monocyte-macrophages purified from antic- Nucleosides and nucleotides play key roles in the replication and oagulanted-blood samples (peripheral blood mononuclear cells) transcription of genetic information, and therefore they have been (Enjuanes et al., 1976) or from pulmonary lavage of swine (Carrascosa utilized in chemotherapy, antibacterial and antiviral therapeutics for et al., 1982). Both sources allow researchers to isolate up to 1 × 109 many decades (Foss, 2000; Yssel et al., 2017). Nucleoside analogues macrophages and use them immediately or store frozen for many years. possess one or more modifications thereby mimicking the structure of These cells could be used to diagnose, grow and titrate the ASFV iso- natural nucleosides enough to be incorporated into the DNA or RNA lates obtained from outbreaks. However, the application of porcine replication cycle