RNA interference-mediated silencing of the respiratory syncytial virus nucleocapsid defines a potent antiviral strategy

Rene Alvarez, Sayda Elbashir, Todd Borland, Ivanka Toudjarska, Philipp Hadwiger, Mathias John, Ingo Roehl, Svetlana Shulga Morskaya, Rick Martinello, Jeffrey Kahn, Mark Van Ranst, Ralph A. Tripp, John P. DeVincenzo, Rajendra Pandey, Martin Maier, Lubomir Nechev, Muthiah Manoharan, Victor Kotelianski, Rachel Meyers

Research output: Contribution to journalArticle

110 Citations (Scopus)

Abstract

We describe the design and characterization of a potent human respiratory syncytial virus (RSV) nucleocapsid gene-specific small interfering RNA (siRNA), ALN-RSV01. In in vitro RSV plaque assays, ALN-RSV01 showed a 50% inhibitory concentration of 0.7 nM. Sequence analysis of primary isolates of RSV showed that the siRNA target site was absolutely conserved in 89/95 isolates, and ALN-RSV01 demonstrated activity against all isolates, including those with single-mismatch mutations. In vivo, intranasal dosing of ALN-RSV01 in a BALB/c mouse model resulted in potent antiviral efficacy, with 2.5- to 3.0-log-unit reductions in RSV lung concentrations being achieved when ALN-RSV01 was administered prophylactically or therapeutically in both single-dose and multidose regimens. The specificity of ALN-RSV01 was demonstrated in vivo by using mismatch controls; and the absence of an immune stimulatory mechanism was demonstrated by showing that nonspecific siRNAs that induce alpha interferon and tumor necrosis factor alpha lack antiviral efficacy, while a chemically modified form of ALN-RSV01 lacking measurable immunostimulatory capacity retained full activity in vivo. Furthermore, an RNA interference mechanism of action was demonstrated by the capture of the site-specific cleavage product of the RSV mRNA via rapid amplification of cDNA ends both in vitro and in vivo. These studies lay a solid foundation for the further investigation of ALN-RSV01 as a novel therapeutic antiviral agent for clinical use by humans.

Original languageEnglish (US)
Pages (from-to)3952-3962
Number of pages11
JournalAntimicrobial Agents and Chemotherapy
Volume53
Issue number9
DOIs
StatePublished - 2009

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Nucleocapsid
Respiratory Syncytial Viruses
RNA Interference
Antiviral Agents
Small Interfering RNA
Human respiratory syncytial virus
Interferon-alpha
Inhibitory Concentration 50
Sequence Analysis
Complementary DNA
Tumor Necrosis Factor-alpha
Lung
Messenger RNA
Mutation
Genes
In Vitro Techniques
Therapeutics

ASJC Scopus subject areas

  • Pharmacology (medical)
  • Pharmacology
  • Infectious Diseases

Cite this

RNA interference-mediated silencing of the respiratory syncytial virus nucleocapsid defines a potent antiviral strategy. / Alvarez, Rene; Elbashir, Sayda; Borland, Todd; Toudjarska, Ivanka; Hadwiger, Philipp; John, Mathias; Roehl, Ingo; Morskaya, Svetlana Shulga; Martinello, Rick; Kahn, Jeffrey; Van Ranst, Mark; Tripp, Ralph A.; DeVincenzo, John P.; Pandey, Rajendra; Maier, Martin; Nechev, Lubomir; Manoharan, Muthiah; Kotelianski, Victor; Meyers, Rachel.

In: Antimicrobial Agents and Chemotherapy, Vol. 53, No. 9, 2009, p. 3952-3962.

Research output: Contribution to journalArticle

Alvarez, R, Elbashir, S, Borland, T, Toudjarska, I, Hadwiger, P, John, M, Roehl, I, Morskaya, SS, Martinello, R, Kahn, J, Van Ranst, M, Tripp, RA, DeVincenzo, JP, Pandey, R, Maier, M, Nechev, L, Manoharan, M, Kotelianski, V & Meyers, R 2009, 'RNA interference-mediated silencing of the respiratory syncytial virus nucleocapsid defines a potent antiviral strategy', Antimicrobial Agents and Chemotherapy, vol. 53, no. 9, pp. 3952-3962. https://doi.org/10.1128/AAC.00014-09
Alvarez, Rene ; Elbashir, Sayda ; Borland, Todd ; Toudjarska, Ivanka ; Hadwiger, Philipp ; John, Mathias ; Roehl, Ingo ; Morskaya, Svetlana Shulga ; Martinello, Rick ; Kahn, Jeffrey ; Van Ranst, Mark ; Tripp, Ralph A. ; DeVincenzo, John P. ; Pandey, Rajendra ; Maier, Martin ; Nechev, Lubomir ; Manoharan, Muthiah ; Kotelianski, Victor ; Meyers, Rachel. / RNA interference-mediated silencing of the respiratory syncytial virus nucleocapsid defines a potent antiviral strategy. In: Antimicrobial Agents and Chemotherapy. 2009 ; Vol. 53, No. 9. pp. 3952-3962.
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AU - John, Mathias

AU - Roehl, Ingo

AU - Morskaya, Svetlana Shulga

AU - Martinello, Rick

AU - Kahn, Jeffrey

AU - Van Ranst, Mark

AU - Tripp, Ralph A.

AU - DeVincenzo, John P.

AU - Pandey, Rajendra

AU - Maier, Martin

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