Antibody-targeted cell fusion

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Abstract

Membrane fusion has many potential applications in biotechnology. Here we show that antibody-targeted cell fusion can be achieved by engineering a fusogenic viral membrane glycoprotein complex. Three different single-chain antibodies were displayed at the extracellular C terminus of the measles hemagglutinin (H) protein, and combinations of point mutations were introduced to ablate its ability to trigger fusion through the native viral receptors CD46 and SLAM. When coexpressed with the measles fusion (F) protein, using plasmid cotransfection or bicistronic adenoviral vectors, the retargeted H proteins could mediate antibody-targeted cell fusion of receptor-negative or receptor-positive index cells with receptor-positive target cells. Adenoviral expression vectors mediating human epidermal growth factor receptor (EGFR)-targeted cell fusion were potently cytotoxic against EGFR-positive tumor cell lines and showed superior antitumor potency against EGFR-positive tumor xenografts as compared with control adenoviruses expressing native (untargeted) or CD38-targeted H proteins.

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Figure 1: Fusogenic properties of mutant H glycoproteins displaying anti-CD38 antibody.
Figure 2: Antibody-targeted cell fusion and cell killing.
Figure 3: Targeted cytoreductive gene therapy using homologous targeted cell fusion.
Figure 4: Adenoviral vectors mediating homologous or heterologous targeted cell fusion.

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Acknowledgements

We thank C.D. James for CHO-EGFR cells, Y. Yanagi for CHO-SLAM cells, J. Schlom for MC38-CEA cells, E. Vitetta for SKOV3ip.1 cells, J.P. Atkinson for CD46 plasmid, J.A. Lust for CD38 scFv, R. Hawkins for CEA scFv and G. Winter for EGFR scFv. We also thank M.J. Federspiel and R.G. Vile for critical reading of the manuscript. This study is supported by the Mayo Foundation, Harold W. Siebens Foundation and NIH grants CA100634-01 and CA90636-01.

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Correspondence to Stephen J Russell.

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Nakamura, T., Peng, K., Vongpunsawad, S. et al. Antibody-targeted cell fusion. Nat Biotechnol 22, 331–336 (2004) doi:10.1038/nbt942

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