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Targeting HSV-1 virions for specific binding to epidermal growth factor receptor-vIII-bearing tumor cells

Cancer Gene Therapy volume 17, pages 655–663 (2010)Cite this article

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Abstract

Oncolytic herpes simplex virus (HSV) vectors have been used in early phase human clinical trials as a therapy for recurrent malignant glioblastoma. This treatment proved safe but limited improvements in patient survival were observed. The potency of these vectors might be enhanced by targeting vector infectivity to tumor cells. Glioma tumors often express a mutant form (vIII) of the epidermal growth factor receptor (EGFR) resulting in the presence of a novel epitope on the cell surface. This epitope is specifically recognized by a single-chain antibody designated MR1-1. HSV-1 infection involves initial binding to heparan sulfate (HS) on the cell surface mediated primarily by the viral envelope, glycoprotein C (gC). Here we joined the MR1-1 single-chain antibody (scFv) to the gC sequence deleted for the HS-binding domain as a means of targeting viral attachment to EGFRvIII on glial tumor cells. Virions bearing MR1-1-modified gC had fivefold increased infectivity for EGFRvIII-bearing human glioma U87 cells compared to mutant receptor-deficient cells. Further, MR1-1/EGFRvIII-mediated infection was more efficient for EGFRvIII-positive cells than was wild-type virus for either positive or negative cells. Sustained infection of EGFRvIII+ glioma cells by MR1-1-modified gC-bearing oncolytic virus, as compared to wild-type gC oncolytic virus, was also shown in subcutaneous tumors in vivo using firefly luciferase as a reporter of infection. These data show that HSV tropism can be manipulated so that virions recognize a cell-specific binding site with increased infectivity for the target cell. The retargeting of HSV infection to tumor cells should enhance vector specificity, tumor cell killing and vector safety.

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Abbreviations

EGFR:

epidermal growth factor receptor

Fluc:

firefly luciferase

gC:

glycoprotein

GFP:

green fluorescent protein

HS:

heparan sulfate

HSBD:

HS-binding domain

HSV:

herpes simplex virus

MOI:

multiplicity of infection

MR1:

antibody to mutant EGFRvIII

PBS:

phosphate-buffered saline

PFU:

plaque-forming unit

RFP:

red fluorescent protein

tu:

transducing unit

X-gal:

5-bromo-4-chloro-3-indolye-β-D-galactopyranoside

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Acknowledgements

We thank Dr Bigner for his kind gift of the MR1-1 and MRB constructs. We also thank Ms Deborah Schuback for advice on virion purification and western blots, Ms Suzanne McDavitt for skilled editorial assistance and the Center for Molecular Imaging Research at Massachusetts General Hospital for help and facilities to assess bioluminescence. This work was supported by NCI Grants CA69246, CA92782 and CA119298, and by NINDS NS 40923.

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Authors and Affiliations

  1. Department of Neurology, Harvard Medical School, Boston, MA, USA

    P Grandi, J Fernandez, D Gianni, M Sena-Esteves & X O Breakefield

  2. Department of Radiology, Harvard Medical School, Boston, MA, USA

    P Grandi, J Fernandez, D Gianni, M Sena-Esteves & X O Breakefield

  3. Department of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    O Szentirmai & R Carter

  4. Neuroscience Program, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    O Szentirmai & R Carter

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Correspondence to P Grandi.

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Grandi, P., Fernandez, J., Szentirmai, O. et al. Targeting HSV-1 virions for specific binding to epidermal growth factor receptor-vIII-bearing tumor cells. Cancer Gene Ther 17, 655–663 (2010). https://doi.org/10.1038/cgt.2010.22

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