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Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2

Nature Medicine volume 5, pages 71–77 (1999)Cite this article

Abstract

Adeno-associated virus 2 (AAV)-based vectors have gained attention as a potentially useful alternative to the more commonly used retroviral and adenoviral vectors for human gene therapy. Although AAV uses the ubiquitously expressed cell surface heparan sulfate proteoglycan (HSPG) as a receptor, the transduction efficiency of AAV vectors varies greatly in different cells and tissues in vitro and in vivo . We demonstrate here that cell surface expression of HSPG alone is insufficient for AAV infection, and that AAV also requires human fibroblast growth factor receptor 1 (FGFR1) as a co-receptor for successful viral entry into the host cell. We document that cells that do not express either HSPG or FGFR1 fail to bind AAV and, consequently, are resistant to infection by AAV. These non-permissive cells are successfully transduced by AAV vectors after stable transfections with cDNAs encoding the murine HSPG and the human FGFR1. Furthermore, AAV infection of permissive cells, known to express both FGFR1 and the epidermal growth factor receptor, is abrogated by treatment of cells with basic fibroblast growth factor, but not with epidermal growth factor. The identification of FGFR1 as a co-receptor for AAV should provide new insights not only into its role in the life cycle of AAV, but also in the optimal use of AAV vectors in human gene therapy.

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Figure 1: Analysis of binding of wild-type AAV to different cell types (a ) and comparative analyses of transduction efficiency of the recombinant vCMVp-lacZ vector in HeLa and NIH3T3 cells (b).
Figure 2: Comparative analyses of binding of 125I-bFGF (a) and 35S-AAV (b) to M07e and Raji cells after either mock transfection or stable transfection with HSPG and/or FGFR1 expression plasmids.
Figure 3: Analysis of transgene expression in M07e (left panels) and Raji (right panels) cells.
Figure 4: Southern blot analysis of the viral DNA entry into cells.
Figure 5: Comparative analyses of transgene expression in M07e (top panels) and Raji (bottom panels) cells stably co-transfected with HSPG and FGFR1 expression plasmids in the presence of co-infection with adenovirus or with prior treatment with tyrphostin 1.
Figure 6: Effect of bFGF and EGF on AAV binding to non-permissive (a) and permissive (b) cells, and on entry into permissive cells (c ).
Figure 7: A model for the role of cell surface HSPG and FGFR1 in mediating AAV binding and entry into the host cell.

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References

  1. Berns, K.I. & Giraud, C. Biology of adeno-associated virus. Curr. Top. Microbiol. Immunol. 218, 1– 23 (1996).

    CAS  PubMed  Google Scholar 

  2. Srivastava, A., Lusby, E.W. & Berns, K.I. Nucleotide sequence and organization of the adeno-associated virus 2 genome. J. Virol. 45, 555– 564 (1983).

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Kotin, R.M., Menninger, J.C., Ward, D.C. & Berns, K.I. Mapping and direct visualization of a region-specific viral DNA integration site on chromosome 19q13-qter. Genomics 10, 831–834 (1991).

    Article  CAS  PubMed  Google Scholar 

  4. Samulski, R.J. et al. Targeted integration of adeno-associated virus (AAV) into human chromosome 19. EMBO J. 10, 3941– 3950 (1991).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Kearns, W.G. et al. Recombinant adeno-associated virus (AAV-CFTR) vectors do not integrate in a site-specific fashion in an immortalized epithelial cell line. Gene Ther. 3, 748–755 (1996).

    CAS  PubMed  Google Scholar 

  6. Ponnazhagan, S. et al. Lack of site-specific integration of the recombinant adeno-associated virus 2 genomes in human cells. Hum. Gene Ther. 8, 275–284 (1997).

    Article  CAS  PubMed  Google Scholar 

  7. Qing, K.Y. et al. Role of tyrosine phosphorylation of a cellular protein in adeno-associated virus 2-mediated transgene expression. Proc. Natl. Acad. Sci. USA 94, 10879–10884 ( 1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Qing, K.Y. et al. Adeno-associated virus type 2-mediated gene transfer: Correlation of tyrosine phosphorylation of the cellular single-stranded D sequence-binding protein with transgene expression in human cells in vitro and murine tissues in vivo. J. Virol. 72, 1593 –1599 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Fisher K.J. et al. Transduction with recombinant adeno-associated virus for gene therapy is limited by leading strand synthesis. J. Virol. 70, 520–532 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Ferrari, F.K., Samulski, T., Shenk, T. & Samulski, R.J. Second-strand synthesis is a rate limiting step for efficient transduction by recombinant adeno-associated virus vectors. J. Virol. 70, 3227–3234 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Muzyczka N. Use of adeno-associated virus as a general transduction vector for mammalian cells. Curr. Top. Microbiol. Immunol. 158, 97–129 (1992).

    CAS  PubMed  Google Scholar 

  12. Summerford, C. & Samulski, R.J. Membrane-associated heparan sulfate proteoglycan is a receptor for adeno-associated virus type 2 virions. J. Virol. 72, 1438– 1445 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Ponnazhagan, S. et al. Adeno-associated virus type 2-mediated transduction of primary human bone marrow-derived CD34+ hematopoietic progenitor cells: Donor variation and correlation of transgene expression with cellular differentiation. J. Virol. 71, 8262–8267 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Ponnazhagan, S. et al. Differential expression from the p6 promoter of human parvovirus B19 in human cells following plasmid transfection and recombinant adeno-associated virus 2 (AAV) infection: Human megakaryocytic leukaemia cells are non-permissive for AAV infection. J. Gen. Virol. 77, 1111 –1122 (1996).

    Article  CAS  PubMed  Google Scholar 

  15. Lebakken, C.S. & Rapraeger, A.C. Syndecan-1 mediates cell spreading in transfected human lymphoblastoid (Raji) cells. J. Cell Biol. 132, 1209– 1221 (1996).

    Article  CAS  PubMed  Google Scholar 

  16. Kiefer, M.C. et al. Ligand-affinity cloning and structure of a cell surface heparan sulfate proteoglycan that binds basic fibroblast growth factor. Proc. Natl. Acad. Sci. USA 87, 6985– 6989 (1990).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Mah, C. et al. Adeno-associated virus 2-mediated gene transfer: Role of epidermal growth factor receptor protein tyrosine kinase in transgene expression. J. Virol. 72, 9835–9843 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Rapraeger, A.C., Krufka, A. & Olwin, B.B. Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation. Science 252, 1705–1708 (1991).

    Article  CAS  PubMed  Google Scholar 

  19. Ledoux, D., Gannoun-Zaki, L. & Barritault D. Interactions of FGFs with target cells. Prog. Growth Factor Res. 4, 107–120 (1992).

    Article  CAS  PubMed  Google Scholar 

  20. Roghani, M. & Moscatelli, D. Basic fibroblast growth factor is internalized through both receptor-mediated and heparan sulfate-mediated mechanisms. J. Biol. Chem. 267, 22156– 22162 (1992).

    CAS  PubMed  Google Scholar 

  21. Givol, D. & Yayon, A. Complexity of FGF receptors: Genetic basis for structural diversity and functional specificity. FASEB J. 6, 3362–3369 ( 1992).

    Article  CAS  PubMed  Google Scholar 

  22. Kan, M. et al. An essential heparin-binding domain in the fibroblast growth factor receptor kinase. Science 259, 1918– 1921 (1993).

    Article  CAS  PubMed  Google Scholar 

  23. Bartlett, J.S. & Samulski, R.J. Fluorescent viral vectors: A new technique for the pharmacological analysis of gene therapy. Nature Med. 4, 635–637 (1998).

    Article  CAS  PubMed  Google Scholar 

  24. Strawn, L.M. et al. Flk-1 as a target for tumor growth inhibition. Cancer Res. 56, 3540–3545 (1996).

    CAS  PubMed  Google Scholar 

  25. Mohammadi, M. et al. Structures of the tyrosine kinase domain of fibroblast growth factor receptor in complex with inhibitors. Science 276, 955–960 (1997).

    Article  CAS  PubMed  Google Scholar 

  26. Ponnazhagan, S. et al. Adeno-associated virus 2-mediated gene transfer in vivo : Organ tropism and expression of transduced sequences in mice. Gene 190, 203–210 ( 1997).

    Article  CAS  PubMed  Google Scholar 

  27. Sheih, H., Montgomery R.I., Esko, J.D. & Spear, P.G. Cell surface receptors for herpes simplex virus are heparan sulfate proteoglycans. J. Cell Biol. 116, 1273–1281 ( 1992).

    Article  Google Scholar 

  28. Laquerre, S. et al. Heparan sulfate proteoglycan binding by herpes simplex virus type 1 glycoproteins B and C, which differ in their contributions to virus attachment, penetration, and cell-to-cell spread. J. Virol. 72, 6119–6130 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Jackson, T. et al. Efficient infection of cells in culture by type O foot-and-mouth disease virus requires binding to cell surface heparan sulfate. J. Virol. 70, 5282–5287 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Chen, Y. et al. Dengue virus infectivity depends on envelope protein binding to target cell heparan sulfate. Nature Med. 3, 866–871 (1997).

    Article  CAS  PubMed  Google Scholar 

  31. Alkhatib, G. et al. CC CKR5: A RANTES, MIP-1a, MIP-1b receptor as a fusion cofactor for macrophage-tropic HIV-1. Science 272, 1955–1958 (1996).

    Article  CAS  PubMed  Google Scholar 

  32. He, J. et al. CCR3 and CCR5 are co-receptors for HIV-1 infection of microglia. Nature 385, 645–649 (1997).

    Article  CAS  PubMed  Google Scholar 

  33. Bergelson, J.M. et al. Isolation of a common receptor for coxsackie B viruses and adenoviruses 2 and 5. Science 275, 1320– 1323 (1997).

    Article  CAS  PubMed  Google Scholar 

  34. Montgomery, R.I., Warner, M.S., Lum, B.J. & Spear, P.G. Herpes Simplex Virus-1 entry into cells mediated by a novel member of the TNF/NGF receptor family. Cell 87, 427–436 (1996).

    Article  CAS  PubMed  Google Scholar 

  35. Geraghty, R.J. et al. Entry of alphaviruses mediated by poliovirus receptor-related protein 1 and poliovirus receptor. Science 280, 1618–1620 (1998).

    Article  CAS  PubMed  Google Scholar 

  36. Green, P.J., Walsh, F.S. & Doherty, P. Promiscuity of fibroblast growth factor receptors. BioEssays 18, 639–646 ( 1996).

    Article  CAS  PubMed  Google Scholar 

  37. Saunders, S., Jalkanen, M., O'Farrell, S. & Bernfield, M. Molecular cloning of syndecan, an integral membrane proteoglycan. J. Cell Biol. 108, 1547–1556 (1989).

    Article  CAS  PubMed  Google Scholar 

  38. Johnson, D.E., Lee, P.L., Lu, J. & Williams, L.T. Diverse forms of a receptor for acidic and basic fibroblast growth factors. Mol. Cell. Biol. 10, 4728–4736 (1990).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Kube, D.M. & Srivastava, A. Quantitative slot-blot analysis: Inhibition of DNA binding to membranes my magnesium ions. Nucl. Acids Res. 25, 3375–3376 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Wang, X.S. et al. Characterization of wild-type adeno-associated virus type 2-like particles generated during recombinant viral vector production and strategies for their elimination. J. Virol. 72, 5472–5480 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank H.E. Broxmeyer for a critical review of this manuscript as well as for his support, and M.-Q. Tan for assistance in some experiments. This research was supported in part by Public Health Service grants (HL-48342, HL-53586, HL-58881, and DK-49218, Centers of Excellence in Molecular Hematology) from the National Institutes of Health, and a grant from the Phi Beta Psi Sorority. A.S. was supported by an Established Investigator Award from the American Heart Association. This manuscript is dedicated to the memories of Anita and Anjali Srivastava.

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Author notes

  1. Shangzhen Zhou, Varavani Dwarki and Arun Srivastava: Virology Department, Chiron Corporation, Emeryville, California 94608, USA K.Q. & C.M. contributed equally to this study

Authors and Affiliations

  1. Departments of Microbiology & Immunology, Indiana University School of Medicine, Indianapolis, 46202, Indiana , USA

    Keyun Qing, Cathryn Mah, Jonathan Hansen & Arun Srivastava

  2. Walther Oncology Center, Indiana University School of Medicine, Indianapolis, 46202, Indiana, USA

    Keyun Qing, Cathryn Mah, Jonathan Hansen & Arun Srivastava

  3. Division of Hematology/Oncology, Department of Medicine Indiana University School of Medicine, Indianapolis, 46202, Indiana, USA

    Arun Srivastava

  4. Walther Cancer Institute, Indianapolis, 46202, Indiana, USA

    Keyun Qing, Cathryn Mah, Jonathan Hansen & Arun Srivastava

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Correspondence to Arun Srivastava.

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Qing, K., Mah, C., Hansen, J. et al. Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2. Nat Med 5, 71–77 (1999). https://doi.org/10.1038/4758

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