Adenovirus-mediated RNA interference against herpes simplex virus infection in vitro
Abstract
Introduction. Herpetic keratitis caused by the herpes simplex virus (HSV) is the most common form of ocular herpes that causes corneal blindness. Although treatments for herpes keratitis have improved in recent years. there is still considerable room for new treatments against viral infection that shows great promise. The aim of the study was to evaluate the effect of RNA interference on HSV Type 1 (HSV1) infection in vitro, first prophylactically then therapeutically.
Material and methods. The highly conserved glycoproteins D (gD) and E (gE) were chosen as targets for this study. Different small interfering RNA (siRNA) duplexes that target gD and gE were designed and chemically synthesized. The recombinant adenovirus type 5 was developed and used as the vehicle with which we delivered the siRNA into the Vero cells infected with the HSV1 KOS strain. Evaluation of the efficacy of siRNA-mediated inhibition was performed either before virus inoculation (prophylactically) or after virus inoculation at the first appearance of lesions (therapeutically). The expression of messenger RNA encoding gD and gE was detected using a real-time polymerase chain reaction (qPCR). We analyzed HSV replication in Vero cells, cytotoxicity of HSV, and cell viability.
Results. When used prophylactically, the siRNA-targeting gD and gE created a more marked decrease in viral titer than when used therapeutically. The transfection of cells with recombinant adenovirus containing the siRNA expression cassette was associated with very low cytotoxicity.
Conclusions. Adenovirus-mediated siRNA-targeting gD and gE genes effectively inhibit the replication of the
HSV in Vero cells. In addition, these findings indicate that the prophylactic use of siRNA is far more effective at inhibiting HSV replication than the therapeutic use.
Keywords: herpetic keratitisHSV infectionVero cellsglycoprotein Dglycoprotein Eadenovirus RNA interferencerecombinant adenovirus type 5cell viability
References
- Giménez F, Suryawanshi A, Rouse BT. Pathogenesis of herpes stromal keratitis--a focus on corneal neovascularization. Prog Retin Eye Res. 2013; 33: 1–9.
- Austin A, Lietman T, Rose-Nussbaumer J. Update on the Management of Infectious Keratitis. Ophthalmology. 2017; 124(11): 1678–1689.
- Azher TN, Yin XT, Tajfirouz D, et al. Herpes simplex keratitis: challenges in diagnosis and clinical management. Clin Ophthalmol. 2017; 11: 185–191.
- Bacon TH, Levin MJ, Leary JJ, et al. Herpes simplex virus resistance to acyclovir and penciclovir after two decades of antiviral therapy. Clin Microbiol Rev. 2003; 16(1): 114–128.
- Piret J, Boivin G. Resistance of herpes simplex viruses to nucleoside analogues: mechanisms, prevalence, and management. Antimicrob Agents Chemother. 2011; 55(2): 459–472.
- Tabbara KF, Al Balushi N. Topical ganciclovir in the treatment of acute herpetic keratitis. Clin Ophthalmol. 2010; 4: 905–912.
- Bondre VP, Sankararaman V, Andhare V, et al. Genetic characterization of human herpesvirus type 1: Full-length genome sequence of strain obtained from an encephalitis case from India. Indian J Med Res. 2016; 144(5): 750–760.
- Heming JD, Conway JF, Homa FL. Herpesvirus Capsid Assembly and DNA Packaging. Adv Anat Embryol Cell Biol. 2017; 223: 119–142.
- Kaye S, Choudhary A. Herpes simplex keratitis. Prog Retin Eye Res. 2006; 25(4): 355–380.
- Montgomery MK, Xu S, Fire A. RNA as a target of double-stranded RNA-mediated genetic interference in Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1998; 95(26): 15502–15507.
- Xu W, Jiang X, Huang L. RNA Interference Technology. Comprehensive Biotechnology. 2019: 560–575.
- Wilson RC, Doudna JA. Molecular mechanisms of RNA interference. Annu Rev Biophys. 2013; 42: 217–239.
- Carmichael GG. Medicine: silencing viruses with RNA. Nature. 2002; 418(6896): 379–380.
- Ui-Tei K, Naito Y, Takahashi F, et al. Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference. Nucleic Acids Res. 2004; 32(3): 936–948.
- Davison AJ. Herpesvirus systematics. Vet Microbiol. 2010; 143(1): 52–69.
- Subak-Sharpe JH, Dargan DJ. HSV molecular biology: general aspects of herpes simplex virus molecular biology. Virus Genes. 1998; 16(3): 239–251.
- Beilstein F, Cohen GH, Eisenberg RJ, et al. Dynamic organization of Herpesvirus glycoproteins on the viral envelope revealed by super-resolution microscopy. PLoS Pathog. 2019; 15(12): e1008209.
- Dubin G, Basu S, Mallory DL, et al. Characterization of domains of herpes simplex virus type 1 glycoprotein E involved in Fc binding activity for immunoglobulin G aggregates. J Virol. 1994; 68(4): 2478–2485.
- Weed DJ, Nicola AV. Herpes simplex virus Membrane Fusion. Adv Anat Embryol Cell Biol. 2017; 223: 29–47.
- Carmichael JC, Yokota H, Craven RC, et al. The HSV-1 mechanisms of cell-to-cell spread and fusion are critically dependent on host PTP1B. PLoS Pathog. 2018; 14(5): e1007054.
- Agelidis AM, Shukla D. Cell entry mechanisms of HSV: what we have learned in recent years. Future Virol. 2015; 10(10): 1145–1154.
- Dingwell KS, Brunetti CR, Hendricks RL, et al. Herpes simplex virus glycoproteins E and I facilitate cell-to-cell spread in vivo and across junctions of cultured cells. J Virol. 1994; 68(2): 834–845.
- Brunetti CR, Dingwell KS, Wale C, et al. Herpes simplex virus gD and virions accumulate in endosomes by mannose 6-phosphate-dependent and -independent mechanisms. J Virol. 1998; 72(4): 3330–3339.
- Burns WH, Saral R, Santos GW, et al. Isolation and characterisation of resistant Herpes simplex virus after acyclovir therapy. Lancet. 1982; 1(8269): 421–423.
- Crumpacker CS, Schnipper LE, Marlowe SI, et al. Resistance to antiviral drugs of herpes simplex virus isolated from a patient treated with acyclovir. N Engl J Med. 1982; 306(6): 343–346.
- Farnsworth A, Goldsmith K, Johnson DC. Herpes simplex virus glycoproteins gD and gE/gI serve essential but redundant functions during acquisition of the virion envelope in the cytoplasm. J Virol. 2003; 77(15): 8481–8494.
- McMillan TN, Johnson DC. Cytoplasmic domain of herpes simplex virus gE causes accumulation in the trans-Golgi network, a site of virus envelopment and sorting of virions to cell junctions. J Virol. 2001; 75(4): 1928–1940.
- Turner A, Bruun B, Minson T, et al. Glycoproteins gB, gD, and gHgL of herpes simplex virus type 1 are necessary and sufficient to mediate membrane fusion in a Cos cell transfection system. J Virol. 1998; 72(1): 873–875.
- Zhe R, Mei-Ying Z, Kitazato K, et al. Effect of siRNA on HSV-1 plaque formation and relative expression levels of UL39 mRNA. Arch Virol. 2008; 153(7): 1401–1406.
- Bhuyan PK, Karikò K, Capodici J, et al. Short interfering RNA-mediated inhibition of herpes simplex virus type 1 gene expression and function during infection of human keratinocytes. J Virol. 2004; 78(19): 10276–10281.
- Norberg P, Bergström T, Rekabdar E, et al. Phylogenetic analysis of clinical herpes simplex virus type 1 isolates identified three genetic groups and recombinant viruses. J Virol. 2004; 78(19): 10755–10764.
- Stingley SW, Ramirez JJ, Aguilar SA, et al. Global analysis of herpes simplex virus type 1 transcription using an oligonucleotide-based DNA microarray. J Virol. 2000; 74(21): 9916–9927.
- Duffy S, Shackelton LA, Holmes EC. Rates of evolutionary change in viruses: patterns and determinants. Nat Rev Genet. 2008; 9(4): 267–276.
- Sarisky RT, Nguyen TT, Duffy KE, et al. Difference in incidence of spontaneous mutations between Herpes simplex virus types 1 and 2. Antimicrob Agents Chemother. 2000; 44(6): 1524–1529.
- Coen DM, Furman PA, Gelep PT, et al. Mutations in the herpes simplex virus DNA polymerase gene can confer resistance to 9-beta-D-arabinofuranosyladenine. J Virol. 1982; 41(3): 909–918.
- Kamp C, Wilke C, Adami C, et al. Viral evolution under the pressure of an adaptive immune system: Optimal mutation rates for viral escape. Complexity. 2003; 8(2): 28–33.
- Gitlin L, Stone JK, Andino R. Poliovirus escape from RNA interference: short interfering RNA-target recognition and implications for therapeutic approaches. J Virol. 2005; 79(2): 1027–1035.
- Kahana R, Kuznetzova L, Rogel A, et al. Inhibition of foot-and-mouth disease virus replication by small interfering RNA. J Gen Virol. 2004; 85(Pt 11): 3213–3217.
- Jin F, Li S, Zheng K, et al. Silencing herpes simplex virus type 1 capsid protein encoding genes by siRNA: a promising antiviral therapeutic approach. PLoS One. 2014; 9(5): e96623.
- Chen W, Yan W, Du Q, et al. RNA interference targeting VP1 inhibits foot-and-mouth disease virus replication in BHK-21 cells and suckling mice. J Virol. 2004; 78(13): 6900–6907.
- Watanabe T, Umehara T, Kohara M. Therapeutic application of RNA interference for hepatitis C virus. Adv Drug Deliv Rev. 2007; 59(12): 1263–1276.
- Coburn GA, Cullen BR. Potent and specific inhibition of human immunodeficiency virus type 1 replication by RNA interference. J Virol. 2002; 76(18): 9225–9231.
