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MicroRNA-based strategy to mitigate the risk of gain-of-function influenza studies

Nature Biotechnology volume 31, pages 844–847 (2013)Cite this article

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Abstract

Recent gain-of-function studies in influenza A virus H5N1 strains revealed that as few as three-amino-acid changes in the hemagglutinin protein confer the capacity for viral transmission between ferrets1,2. As transmission between ferrets is considered a surrogate indicator of transmissibility between humans, these studies raised concerns about the risks of gain-of-function influenza A virus research. Here we present an approach to strengthen the biosafety of gain-of-function influenza experiments. We exploit species-specific endogenous small RNAs to restrict influenza A virus tropism. In particular, we found that the microRNA miR-192 was expressed in primary human respiratory tract epithelial cells as well as in mouse lungs but absent from the ferret respiratory tract. Incorporation of miR-192 target sites into influenza A virus did not prevent influenza replication and transmissibility in ferrets, but did attenuate influenza pathogenicity in mice. This molecular biocontainment approach should be applicable beyond influenza A virus to minimize the risk of experiments involving other pathogenic viruses.

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Figure 1: Species-specific miRNA expression.
Figure 2: Species-specific miRNAs can be exploited to control influenza A virus tropism.
Figure 3: Species-specific miR-192 targeting attenuates influenza A virus infection in mice.
Figure 4: Species-specific miRNA targeting of influenza A virus does not affect virus capacity to infect, replicate or transmit between ferrets.

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Acknowledgements

This work was supported by the Center for Research on Influenza Pathogenesis, a National Institute of Allergy and Infectious Diseases–funded Center of Excellence in Influenza Research and Surveillance (HHSN266200700010C). B.R.T. is supported in part by the US Army Research Office under grant numbers W911NF-12-R-0012 and W911NF-08-1-0413 and the Burroughs Wellcome Fund. MDCK cells were kindly provided by P. Palese (Mount Sinai School of Medicine).

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Author notes

  1. Ryan A Langlois and Randy A Albrecht: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA

    Ryan A Langlois, Randy A Albrecht, Jillian S Shapiro, Mark A Chua, Adolfo García-Sastre & Benjamin R tenOever

  2. Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA

    Ryan A Langlois, Randy A Albrecht, Adolfo García-Sastre & Benjamin R tenOever

  3. Department of Veterinary Medicine, University of Maryland, College Park, Maryland, USA

    Brian Kimble, Troy Sutton, Courtney Finch, Matthew Angel, Ana Silvia Gonzalez-Reiche, Kemin Xu & Daniel Perez

  4. Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, New York, USA

    Adolfo García-Sastre & Benjamin R tenOever

Authors
  1. Ryan A Langlois
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  2. Randy A Albrecht
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  11. Daniel Perez
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Contributions

R.A.L. and R.A.A. designed and conducted the research and wrote the manuscript. B.K., T.S., C.F., A.S.G.-R., M.A. and K.X. were responsible for the transmission experiments. J.S.S. performed and analyzed deep sequencing experiments. M.A.C. provided technical support. D.P., A.G.-S. and B.R.T. designed research, oversaw the project and wrote the manuscript.

Corresponding authors

Correspondence to Daniel Perez, Adolfo García-Sastre or Benjamin R tenOever.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4 (PDF 330 kb)

Supplementary Table 1

miRNA expression in A549, MDCK and ferret lung cells (XLSX 57 kb)

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Langlois, R., Albrecht, R., Kimble, B. et al. MicroRNA-based strategy to mitigate the risk of gain-of-function influenza studies. Nat Biotechnol 31, 844–847 (2013). https://doi.org/10.1038/nbt.2666

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