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Peptide nanofibrils boost retroviral gene transfer and provide a rapid means for concentrating viruses

Abstract

Inefficient gene transfer and low virion concentrations are common limitations of retroviral transduction1. We and others have previously shown that peptides derived from human semen form amyloid fibrils that boost retroviral gene delivery by promoting virion attachment to the target cells2,3,4,5,6,7,8. However, application of these natural fibril-forming peptides is limited by moderate efficiencies, the high costs of peptide synthesis, and variability in fibril size and formation kinetics. Here, we report the development of nanofibrils that self-assemble in aqueous solution from a 12-residue peptide, termed enhancing factor C (EF-C). These artificial nanofibrils enhance retroviral gene transfer substantially more efficiently than semen-derived fibrils or other transduction enhancers. Moreover, EF-C nanofibrils allow the concentration of retroviral vectors by conventional low-speed centrifugation, and are safe and effective, as assessed in an ex vivo gene transfer study. Our results show that EF-C fibrils comprise a highly versatile, convenient and broadly applicable nanomaterial that holds the potential to significantly facilitate retroviral gene transfer in basic research and clinical applications.

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Figure 1: Peptides derived from the HIV-1 glycoprotein gp120 enhance virus infection.
Figure 2: Structural characterization and molecular modelling of EF-C fibrils.
Figure 3: EF-C fibrils bind, precipitate and concentrate virions.
Figure 4: EF-C fibrils are broad-based transduction enhancers.
Figure 5: EF-C fibrils can be immobilized (a–c) and allow efficient gene transfer into mice (d–f).

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Acknowledgements

The authors thank S. Liu for support in the initial phase of the project and D. Krnavek and M. Schlaier for expert technical assistance. The authors also thank W. Mothes for providing the MLV GAG-CFP plasmid, B. Böhm for Bon cells, J. von Einem for HFF cells, the AIDS Research and Reference Program for U87-MG and TZM-bl cells, and N. Landau for CEMx-M7 (CEMx174 5.25 M7) cells. This work was supported by a grant from the German Research Foundation to J.M. Molecular simulations were performed using the resources of Moscow University Supercomputing Center.

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Contributions

M.Y. generated virus stocks and performed most transductions. C.M. and T.W. assisted in writing the manuscript and were responsible for the structural elucidation of EF-C fibrils. A.K.S., P.G.K. and A.R.K. performed molecular simulations. V.V. and H.G. conducted the ex vivo gene transfer study. C.W.B. and X.S. performed CD analysis. F.A. and D.P. performed coating experiments and measured zeta potentials. O.Z. and J.M. performed Congo Red and ThT assays, S.M.U. was responsible for the analysis of immobilized EF-C by confocal microscopy. D.S. and C.G. performed the fusion assay and flow cytometry assays. P.W. assisted in microscopy. O.L. and T.S. studied the EF-C interaction with virions and cells. J.B. provided retro- and lentiviral vectors and knowhow. H.S. and K.S. isolated and provided human stem cells and performed colony-forming unit assays. L.S. and W.G.F. synthesized and provided peptides. N.R.R. assisted in planning and writing. T.K. analysed AFM data. F.K. and J.M. planned research, analysed data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Jan Münch.

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Competing interests

M. Yo., F. Ki. and J. Mü. filed for a patent to use EF-C fibrils as tranduction and infection enhancer.

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Yolamanova, M., Meier, C., Shaytan, A. et al. Peptide nanofibrils boost retroviral gene transfer and provide a rapid means for concentrating viruses. Nature Nanotech 8, 130–136 (2013). https://doi.org/10.1038/nnano.2012.248

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