title: Small molecule inhibitors of Ago2 decrease Venezuelan equine encephalitis virus replication.
abstract: Venezuelan equine encephalitis virus (VEEV) is classified as a Category B Select Agent and potential bioterror weapon for its severe disease course in humans and equines and its potential for aerosol transmission. There are no current FDA licensed vaccines or specific therapies against VEEV, making identification of potential therapeutic targets a priority. With this aim, our research focuses on the interactions of VEEV with host microRNA (miRNA) machinery. miRNAs are small non-coding RNAs that act as master regulators of gene expression by downregulating or degrading messenger RNA, thus suppressing production of the resultant proteins. Recent publications implicate miRNA interactions in the pathogenesis of various viral diseases. To test the importance of miRNA processing for VEEV replication, cells deficient in Ago2, an important component of the RNA-induced silencing complex (RISC), and cells treated with known Ago2 inhibitors, notably acriflavine (ACF), were utilized. Both conditions caused decreased viral replication and capsid expression. ACF treatment promoted increased survival of neuronal cells over a non-treated, infected control and reduced viral titers of fully virulent VEEV as well as Eastern and Western Equine Encephalitis Viruses and West Nile Virus, but not Vesicular Stomatitis Virus. ACF treatment of VEEV TC-83 infected mice resulted in increased in vivo survival, but did not affect survival or viral loads when mice were challenged with fully virulent VEEV TrD. These results suggest that inhibition of Ago2 results in decreased replication of encephalitic alphaviruses in vitro and this pathway may be an avenue to explore for future therapeutic development.
authors.affiliation: National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, USA.
authors.forename: Cathaleen
authors.initials: C
authors.lastname: Madsen
authors.affiliation: National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, USA.
authors.forename: Idris
authors.initials: I
authors.lastname: Hooper
authors.affiliation: National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, USA.
authors.forename: Lindsay
authors.initials: L
authors.lastname: Lundberg
authors.affiliation: National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, USA.
authors.forename: Nazly
authors.initials: N
authors.lastname: Shafagati
authors.affiliation: National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, USA.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Johnson
authors.affiliation: National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, USA.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Senina
authors.affiliation: National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, USA.
authors.forename: Cynthia
authors.initials: C
authors.lastname: de la Fuente
authors.affiliation: Maryland Pathogen Research Institute, College Park, MD, 20742, USA; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, 20742, USA.
authors.forename: Lisa I
authors.initials: LI
authors.lastname: Hoover
authors.affiliation: Maryland Pathogen Research Institute, College Park, MD, 20742, USA; Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, 20742, USA.
authors.forename: Brenda L
authors.initials: BL
authors.lastname: Fredricksen
authors.affiliation: Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, 20742, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Dinman
authors.affiliation: MRI Global, 1330 Piccard Drive #101, Rockville, MD, 20850, USA.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Jacobs
authors.affiliation: National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, USA. Electronic address: kkehnhal@gmu.edu.
authors.forename: Kylene
authors.initials: K
authors.lastname: Kehn-Hall
mesh_terms: D000167:Acriflavine / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology* / Q000627:therapeutic use; D060565:Argonaute Proteins / Q000037:antagonists & inhibitors*; D036022:Capsid Proteins / Q000096:biosynthesis; D002470:Cell Survival; D004195:Disease Models, Animal; D004666:Encephalitis Virus, Venezuelan Equine / Q000187:drug effects* / Q000502:physiology*; D004685:Encephalomyelitis, Venezuelan Equine / Q000188:drug therapy / Q000821:virology; D004791:Enzyme Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D009474:Neurons / Q000502:physiology / Q000821:virology; D016019:Survival Analysis; D016896:Treatment Outcome; D019562:Viral Load; D014779:Virus Replication / Q000187:drug effects*
chemical_list: C526735:Ago2 protein, mouse; D000998:Antiviral Agents; D060565:Argonaute Proteins; D036022:Capsid Proteins; D004791:Enzyme Inhibitors; D000167:Acriflavine
keywords: Acriflavine; Ago2; Alphavirus; Therapeutic; Venezuelan equine encephalitis virus; miRNA