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

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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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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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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