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Nanostructured glycan architecture is important in the inhibition of influenza A virus infection

Nature Nanotechnology volume 12, pages 48–54 (2017)Cite this article

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Abstract

Rapid change1 and zoonotic transmission to humans2 have enhanced the virulence of the influenza A virus (IAV)3. Neutralizing antibodies fail to provide lasting protection from seasonal epidemics1,4. Furthermore, the effectiveness of anti-influenza neuraminidase inhibitors has declined because of drug resistance5. Drugs that can block viral attachment and cell entry independent of antigenic evolution or drug resistance might address these problems. We show that multivalent 6′-sialyllactose-polyamidoamine (6SL–PAMAM) conjugates, when designed to have well-defined ligand valencies and spacings, can effectively inhibit IAV infection. Generation 4 (G4) 6SL–PAMAM conjugates with a spacing of around 3 nm between 6SL ligands (S3–G4) showed the strongest binding to a hemagglutinin trimer (dissociation constant of 1.6 × 10−7 M) and afforded the best inhibition of H1N1 infection. S3–G4 conjugates were resistant to hydrolysis by H1N1 neuraminidase. These conjugates protected 75% of mice from a lethal challenge with H1N1 and prevented weight loss in infected animals. The structure-based design of multivalent nanomaterials, involving modulation of nanoscale backbone structures and number and spacing between ligands, resulted in optimal inhibition of IAV infection. This approach may be broadly applicable for designing effective and enduring therapeutic protection against human or avian influenza viruses.

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Figure 1: Design and synthesis of multivalent 6SL–PAMAM dendrimer conjugates.
Figure 2: The ligand number calculated for the S3–G4 6SL dendrimer conjugates provides for the detailed structure shown.
Figure 3: In vitro inhibitory activity of 6SL–PAMAM dendrimer conjugates against H1N1 virus infection.
Figure 4: In vivo inhibitory efficacy of S3–G4 against H1N1 and HA binding models.

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References

  1. Smith, D. J. et al. Mapping the antigenic and genetic evolution of influenza virus. Science 305, 371–376 (2004).

    Article  CAS  Google Scholar 

  2. Lam, T. T. et al. Dissemination, divergence and establishment of H7N9 influenza viruses in China. Nature 522, 102–105 (2015).

    Article  CAS  Google Scholar 

  3. Schrauwen, E. J. et al. Determinants of virulence of influenza A virus. Eur. J. Clin. Microbiol. Infect. Dis. 33, 479–490 (2014).

    Article  CAS  Google Scholar 

  4. Schrauwen, E. J. & Fouchier, R. A. Host adaptation and transmission of influenza A viruses in mammals. Emerg. Microbes Infect. 3, e9 (2014).

    Article  Google Scholar 

  5. Moscona, A. Global transmission of oseltamivir-resistant influenza. N. Engl. J. Med. 360, 953–956 (2009).

    Article  CAS  Google Scholar 

  6. Shi, Y., Wu, Y., Zhang, W., Qi, J. & Gao, G. F. Enabling the ‘host jump’: structural determinants of receptor-binding specificity in influenza A viruses. Nat. Rev. Microbiol. 12, 822–831 (2014).

    Article  CAS  Google Scholar 

  7. Hai, R. et al. Influenza A(H7N9) virus gains neuraminidase inhibitor resistance without loss of in vivo virulence or transmissibility. Nat. Commun. 4, 2854 (2013).

    Article  Google Scholar 

  8. Rogers, G. N., Pritchett, T. J., Lane, J. L. & Paulson, J. C. Differential sensitivity of human, avian, and equine influenza A viruses to a glycoprotein inhibitor of infection: selection of receptor specific variants. Virology 131, 394–408 (1983).

    Article  CAS  Google Scholar 

  9. Hendricks, G. L. et al. Sialylneolacto-N-tetraose c (LSTc)-bearing liposomal decoys capture influenza A virus. J. Biol. Chem. 288, 8061–8073 (2013).

    Article  CAS  Google Scholar 

  10. Takemoto, D. K., Skehel, J. J. & Wiley, D. C. A surface plasmon resonance assay for the binding of influenza virus hemagglutinin to its sialic acid receptor. Virology 217, 452–458 (1996).

    Article  CAS  Google Scholar 

  11. Reuter, J. D. et al. Inhibition of viral adhesion and infection by sialic-acid-conjugated dendritic polymers. Bioconjug. Chem. 10, 271–278 (1999).

    Article  CAS  Google Scholar 

  12. Gambaryan, A. S. et al. Polymeric inhibitor of influenza virus attachment protects mice from experimental influenza infection. Antiviral Res. 55, 201–205 (2002).

    Article  CAS  Google Scholar 

  13. Esfand, R. & Tomalia, D. A. Poly(amidoamine) (PAMAM) dendrimers: from biomimicry to drug delivery and biomedical applications. Drug Discov. Today 6, 427–436 (2001).

    Article  CAS  Google Scholar 

  14. Goodwin, A. P., Lam, S. S. & Fréchet, JMJ . Rapid, efficient synthesis of heterobifunctional biodegradable dendrimers. J. Am. Chem. Soc. 129, 6994–6995 (2007).

    Article  CAS  Google Scholar 

  15. Mammen, M., Choi, S. K. & Whitesides, G. M. Polyvalent interactions in biological systems: implications for design and use of multivalent ligands and inhibitors. Angew. Chem. Int. Ed. 37, 2754–2794 (1998).

    Article  Google Scholar 

  16. Patke, S. et al. Design of monodisperse and well-defined polypeptide-based polyvalent inhibitors of anthrax toxin. Angew. Chem. Int. Ed. 53, 8037–8040 (2014).

    Article  CAS  Google Scholar 

  17. Glick, G. D., Toogood, P. L., Wiley, D. C., Skehel, J. J. & Knowles, J. R. Ligand recognition by influenza virus: the binding of bivalent sialosides. J. Biol. Chem. 266, 23660–23669 (1991).

    CAS  Google Scholar 

  18. Chinarev, A. A. et al. in Sialobiology and Other Novel Forms of Glycosylation (eds Inoue, Y., Lee, Y. C. & Troy, F. A.) 135–143 (Gakushin, 1999).

    Google Scholar 

  19. Olson, S. T. & Shore, J. D. Binding of high affinity heparin to antithrombin III: characterization of the protein fluorescence enhancement. J. Biol. Chem. 256, 11065–11072 (1981).

    CAS  Google Scholar 

  20. Harris, A. et al. Influenza virus pleiomorphy characterized by cryoelectron tomography. Proc. Natl Acad. Sci. USA 103, 19123–19127 (2006).

    Article  CAS  Google Scholar 

  21. Zhou, Z. H. Towards atomic resolution structural determination by single-particle cryo-electron microscopy. Curr. Opin. Struct. Biol. 18, 218–228 (2008).

    Article  CAS  Google Scholar 

  22. Eichman, J. D., Bielinska, A. U., Kukowska-Latallo, J. F. & Baker, J. R. Jr The use of PAMAM dendrimers in the efficient transfer of genetic material into cells. Pharm. Sci. Technol. Today 3, 232–245 (2000).

    Article  CAS  Google Scholar 

  23. Sakai, T. et al. Dual wavelength imaging allows analysis of membrane fusion of influenza virus inside cells. J. Virol. 80, 2013–2018 (2006).

    Article  CAS  Google Scholar 

  24. Kwon, D., Shin, K., Kim, S. J., Lee, J. Y. & Kang, C. Mammalian pathogenesis of oseltamivir-resistant pandemic (H1N1) 2009 influenza virus isolated in South Korea. Virus Res. 185, 41–46 (2014).

    Article  CAS  Google Scholar 

  25. Matrosovich, M. & Klenk, H. D. Natural and synthetic sialic acid-containing inhibitors of influenza virus receptor binding. Rev. Med. Virol. 13, 85–97 (2003).

    Article  CAS  Google Scholar 

  26. Polizzotti, B. D., Maheshwari, R., Vinkenborg, J. & Kiick, K. L. Effects of saccharide spacing and chain extension on toxin inhibition by glycopolypeptides of well-defined architecture. Macromolecules 40, 7103–7110 (2007).

    Article  CAS  Google Scholar 

  27. Choi, S. K., Mammen, M. & Whitesides, G. M. Monomeric inhibitors of influenza neuraminidase enhance the hemagglutination inhibition activities of polyacrylamides presenting multiple C-sialoside groups. Chem. Biol. 3, 97–104 (1996).

    Article  CAS  Google Scholar 

  28. Sauter, N. K. et al. Binding of influenza virus hemagglutinin to analogs of its cell-surface receptor, sialic acid: analysis by proton nuclear magnetic resonance spectroscopy and X-ray crystallography. Biochemistry 31, 9609–9621 (1992).

    Article  CAS  Google Scholar 

  29. Rowe, T. et al. Detection of antibody to avian influenza A (H5N1) virus in human serum by using a combination of serologic assays. J. Clin. Microbiol. 37, 937–943 (1999).

    CAS  Google Scholar 

  30. Isaacs, C. E. et al. Epigallocatechin gallate inactivates clinical isolates of herpes simplex virus. Antimicrob. Agents Chemother. 52, 962–970 (2008).

    Article  CAS  Google Scholar 

  31. Jung, H. S., Komatsu, S., Ikebe, M. & Craig, R. Head-head and head-tail interaction: a general mechanism for switching off myosin II activity in cells. Mol. Biol. Cell. 19, 3234–3242 (2008).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Research Foundation of Korea grant funded by the Korea government (MSIP) (NRF-2013R1A2A2A01068858), Marine Biotechnology Program (PJT200620) funded by Ministry of Oceans and Fisheries, and Basic Science Research Program funded by the Ministry of Education (2015R1D1A1A01060512).

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  1. Seok-Joon Kwon, Dong Hee Na and Jong Hwan Kwak: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Chemical and Biological Engineering and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Biotech 4005, 110 8th St., Troy, 12180, New York, USA

    Seok-Joon Kwon, Marc Douaisi, Fuming Zhang, Ravi S. Kane, Jonathan S. Dordick & Robert J. Linhardt

  2. College of Pharmacy and Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, 702-701, Republic of Korea

    Dong Hee Na & Eun Ji Park

  3. School of Pharmacy, Sungkyunkwan University, Suwon, 440-746, Republic of Korea

    Jong Hwan Kwak

  4. Laboratory Animal Medicine, College of Veterinary Medicine, Chonnam National University, Gwangju, 500-757, Republic of Korea

    Jong-Hwan Park

  5. Department of Biochemistry, College of Medicine, Konyang University, Daejeon, 302-718, Republic of Korea

    Jong-Hwan Park & Kyung Bok Lee

  6. College of Veterinary Medicine, Konkuk University, Seoul, 143-701, Republic of Korea

    Hana Youn & Chang-Seon Song

  7. Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Biotech 4005, 110 8th St., Troy, 12180, New York, USA

    Robert J. Linhardt

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  1. Seok-Joon Kwon
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Contributions

S.-J.K, D.H.N., and J.H.K. performed all of the experiments and analysed all data. M.D. performed the microneutralization assays. F.Z. performed the SPR analyses. E.J.P. synthesized the 6SL–PAMAM dendrimer conjugates. J.-H.P., H.Y. and C.-S.S. conducted the murine experiments. R.S.K. and J.S.D. provided critical feedback on the manuscript. S.-J.K and K.B.L. planned the experiments, interpreted the results, and wrote the manuscript with R.J.L. and J.S.D.

Corresponding authors

Correspondence to Kyung Bok Lee or Robert J. Linhardt.

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The authors declare no competing financial interests.

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Kwon, SJ., Na, D., Kwak, J. et al. Nanostructured glycan architecture is important in the inhibition of influenza A virus infection. Nature Nanotech 12, 48–54 (2017). https://doi.org/10.1038/nnano.2016.181

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