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. 2018 Jun 20;218(2):218-227.
doi: 10.1093/infdis/jiy152.

Fusion Inhibitory Lipopeptides Engineered for Prophylaxis of Nipah Virus in Primates

Cyrille Mathieu  1   2   3 , Matteo Porotto  1   2 , Tiago N Figueira  1   2   4 , Branka Horvat  3 , Anne Moscona  1   5   6   2
Affiliations

Fusion Inhibitory Lipopeptides Engineered for Prophylaxis of Nipah Virus in Primates

Cyrille Mathieu et al. J Infect Dis. .

Abstract

Background: The emerging zoonotic paramyxovirus Nipah virus (NiV) causes severe respiratory and neurological disease in humans, with high fatality rates. Nipah virus can be transmitted via person-to-person contact, posing a high risk for epidemic outbreaks. However, a broadly applicable approach for human NiV outbreaks in field settings is lacking.

Methods: We engineered new antiviral lipopeptides and analyzed in vitro fusion inhibition to identify an optimal candidate for prophylaxis of NiV infection in the lower respiratory tract, and we assessed antiviral efficiency in 2 different animal models.

Results: We show that lethal NiV infection can be prevented with lipopeptides delivered via the respiratory route in both hamsters and nonhuman primates. By targeting retention of peptides for NiV prophylaxis in the respiratory tract, we avoid its systemic delivery in individuals who need only prevention, and thus we increase the safety of treatment and enhance utility of the intervention.

Conclusions: The experiments provide a proof of concept for the use of antifusion lipopeptides for prophylaxis of lethal NiV. These results advance the goal of rational development of potent lipopeptide inhibitors with desirable pharmacokinetic and biodistribution properties and a safe effective delivery method to target NiV and other pathogenic viruses.

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Figures

Figure 1.
Figure 1.
Influence of lipid moiety on the inhibition of Nipah virus (NiV) G/F-mediated fusion, and protease sensitivity by VIKI series C-terminal heptad-repeat region peptides. (A) Fusion of NiV G/F-coexpressing cells with 293T cells in the presence of serial dilutions of VIKI-dPEG4, VIKI-dPEG4-Chol, VIKI-dPEG4-Toco, VIKI-dPEG4-bisChol, and VIKI-dPEG4-bisToco was quantified at 4 hours, using a β-galactosidase complementation assay. Results are presented as percentage reduction in luminescence (y-axis) compared with no treatment. Each point is the mean ± standard deviation (s.d.) of results with n = 3 experiments. (B) The indicated peptides (30 µM) were incubated with proteinase K (10 µg/mL) at 37°C and collected for analysis at multiple time points, from 0 to 60 minutes. The products of the reaction were subjected to nonreducing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and silver stained. Peptide and proteinase K controls were included in each SDS-PAGE and are shown in the original images, included in Supplementary Figure S2. The intact peptide content in each sample was calculated from densitometry measurements of silver-stained peptide bands in gel images, normalized to the control in the absence of peptide degradation. A single exponential decay curve was fitted by nonlinear regression to the experimental data sets to determine the respective peptide half-life. The data points are the average of 2 independent replicates, and error bars correspond to the s.d.
Figure 2.
Figure 2.
Peptide inhibition of Nipah virus (NiV) infection in hamsters. (A) Hamsters were infected intranasally (i.n.) with 104 or 106 plaque-forming units (pfu) in either 40 or 100 µL Dulbecco’s modified Eagle’s medium or mock infected. Animals were observed for 3 weeks. (B) Peptides were given i.n. to hamsters from day −1 to +1; on day 0, peptides were given concurrently with 106 pfu of NiV. Untreated animals received the vehicle alone. Animals were observed for 3 weeks. Both peptides significantly improved survival (**, P = .007 for VIKI-dPEG4-Toco and *, P = .02 for VIKI-dPEG4-Chol, using a Mantel-Cox test).
Figure 3.
Figure 3.
Biodistribution of VIKI-dPEG4-Chol and VIKI-dPEG4-Toco peptides after intratracheal (i.t.) or i.t. plus subcutaneous (s.c.) administration in African green monkeys (AGMs). African Green Monkeys were administered 10 mg/kg of peptide either i.t. (animals A and B) or 10 mg/kg i.t. plus 2 mg/kg s.c. (animals C and D). (A) At the indicated time points, the peptide concentration was quantitated in plasma (n = 2/data point). The ordinate values represent means (±standard deviation) of results from 2 animals. (B and C) After 14 days, the animals received an additional administration, then sacrificed after 24 hours, and the peptide concentrations in brain and lung were determined by enzyme-linked immunosorbent assay.
Figure 4.
Figure 4.
Peptide inhibition of Nipah virus (NiV) infection in African green monkeys (AGMs). (A) Schematic representation of the experiment. All animals received daily intratracheal (i.t.) administration of VIKI-dPEG4-Toco peptide or vehicle from day −1 to day 5 postinfection, whereas the additional 3 animals were also treated subcutaneously (s.c.) until day 10 (i.t. + s.c.). Animals were infected with 2 × 107 plaque-forming units of NiV given i.t. at day 0, 4 hours before peptide administration. A control uninfected animal received Dulbecco’s modified Eagle’s medium (mock). Three untreated AGMs were administered vehicle alone. Animals were observed for 4 weeks. (B) Peptide administration in both treated groups led to 33% survival (2 of 6 animals). (C) At indicated days, the peptide concentration in plasma was quantitated by enzyme-linked immunosorbent assay in the surviving animals, and the results are presented as means (±standard deviation). (D) The production of neutralizing antibodies was quantitated at the end of the protocol using an NiV-specific seroneutralizing assay.
Figure 5.
Figure 5.
Lymphopenia and monocytosis in African green monkeys (AGMs) after Nipah virus (NiV) infection. Graphs present the evolution of the percentage of circulating lymphocytes (A–C) and monocytes (D–F) over 30 days in AGMs infected with NiV. (A and D) Uninfected treated animal shown with the infected but not treated (NT) animals; (B and E) uninfected treated animal shown with NiV-infected animals treated intratracheally (i.t.) + subcutaneouss.c.; (C and F) uninfected treated animal shown with NiV-infected animals treated only i.t.
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References

    1. Mathieu C, Horvat B. Henipavirus pathogenesis and antiviral approaches. Expert Rev Anti Infect Ther 2015; 13:343–54. - PubMed
    1. Drexler JF, Corman VM, Müller MA et al. . Bats host major mammalian paramyxoviruses. Nat Commun 2012; 3:796. - PMC - PubMed
    1. Marsh GA, de Jong C, Barr JA et al. . Cedar virus: a novel Henipavirus isolated from Australian bats. PLoS Pathog 2012; 8:e1002836. - PMC - PubMed
    1. Luby SP. The pandemic potential of Nipah virus. Antiviral Res 2013; 100:38–43. - PubMed
    1. Middleton D, Pallister J, Klein R et al. . Hendra virus vaccine, a one health approach to protecting horse, human, and environmental health. Emerg Infect Dis 2014; 20:372–9. - PMC - PubMed

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