title: Inhibition of dengue virus infections in cell cultures and in AG129 mice by a small interfering RNA targeting a highly conserved sequence.
abstract: The dengue viruses (DENVs) exist as numerous genetic strains that are grouped into four antigenically distinct serotypes. DENV strains from each serotype can cause severe disease and threaten public health in tropical and subtropical regions worldwide. No licensed antiviral agent to treat DENV infections is currently available, and there is an acute need for the development of novel therapeutics. We found that a synthetic small interfering RNA (siRNA) (DC-3) targeting the highly conserved 5' cyclization sequence (5'CS) region of the DENV genome reduced, by more than 100-fold, the titers of representative strains from each DENV serotype in vitro. To determine if DC-3 siRNA could inhibit DENV in vivo, an "in vivo-ready" version of DC-3 was synthesized and tested against DENV-2 by using a mouse model of antibody-dependent enhancement of infection (ADE)-induced disease. Compared with the rapid weight loss and 5-day average survival time of the control groups, mice receiving the DC-3 siRNA had an average survival time of 15 days and showed little weight loss for approximately 12 days. DC-3-treated mice also contained significantly less virus than control groups in several tissues at various time points postinfection. These results suggest that exogenously introduced siRNA combined with the endogenous RNA interference processing machinery has the capacity to prevent severe dengue disease. Overall, the data indicate that DC-3 siRNA represents a useful research reagent and has potential as a novel approach to therapeutic intervention against the genetically diverse dengue viruses.
authors.affiliation: Vaccine and Gene Therapy Institute, Oregon Health and Science University, Beaverton, OR, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Stein
authors.forename: Stuart T
authors.initials: ST
authors.lastname: Perry
authors.forename: Michael D
authors.initials: MD
authors.lastname: Buck
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Oehmen
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Fischer
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Poore
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Smith
authors.forename: Alissa M
authors.initials: AM
authors.lastname: Lancaster
authors.forename: Alec J
authors.initials: AJ
authors.lastname: Hirsch
authors.forename: Mark K
authors.initials: MK
authors.lastname: Slifka
authors.forename: Jay A
authors.initials: JA
authors.lastname: Nelson
authors.forename: Sujan
authors.initials: S
authors.lastname: Shresta
authors.forename: Klaus
authors.initials: K
authors.lastname: Früh
mesh_terms: D000818:Animals; D019067:Antibody-Dependent Enhancement; D000998:Antiviral Agents / Q000008:administration & dosage* / Q000494:pharmacology*; D001688:Biological Products / Q000008:administration & dosage / Q000494:pharmacology; D001835:Body Weight; D018929:Cell Culture Techniques; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D003715:Dengue / Q000188:drug therapy* / Q000473:pathology / Q000821:virology; D003716:Dengue Virus / Q000187:drug effects* / Q000235:genetics; D004195:Disease Models, Animal; D006801:Humans; D051379:Mice; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics / Q000494:pharmacology*; D012376:Rodent Diseases / Q000188:drug therapy / Q000473:pathology / Q000821:virology; D016019:Survival Analysis
chemical_list: D000998:Antiviral Agents; D001688:Biological Products; D034741:RNA, Small Interfering