Springer Protocols: Abstract: 8. Fiber-modified Adenoviruses for Targeted Gene Therapy

8. Fiber-modified Adenoviruses for Targeted Gene Therapy

By: Hongju Wu³, David T. Curiel³

Abstract

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Human adenovirus serotype 5 (Ad5) has been widely explored as a gene delivery vector. To achieve highly efficient and specific gene delivery, it is often necessary to re-direct Ad5 tropism. Because the capsid protein fiber plays an essential role in directing Ad5 infection, our laboratory attempted to re-target Ad5 through fiber modification. We have developed two strategies in this regard. One is a bi-specific adaptor protein strategy, in which the adaptor protein is designed to bind both the Ad5 fiber and an alternative cell-surface receptor. Another is genetic modification, in which alternative targeting motifs are genetically incorporated into the fiber knob domain so that the Ad5 vectors can infect cells through the alternative receptors. In this chapter, we will focus on the genetic fiber modification strategy and provide a detailed protocol for generation of fiber-modified Ad5 vectors. A series of techniques/procedures used in our laboratory will be described, which include the generation of fiber-modified Ad5 genome by homologous recombination in a bacterial system, rescuing the modified Ad5 viruses, virus amplification and purification, and virus titration.

Affiliation(s): (3) Division of Human Gene Therapy, Department of Obstetrics and Gynecology, and the Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL

Book Title: Gene Therapy Protocols: Design and Characterization of Gene Transfer Vectors

Series: Methods in Molecular Biology | Volume: 434 | Pub. Date: Mar-07-2008 | Page Range: 113-132 | DOI: 10.1007/978-1-60327-248-3_8

Subject: Genetics/Genomics

Key Words: Gene therapy - adenovirus - fiber modification - homologous recombination - rescuing adenoviral vectors - adenovirus amplification - CsCl gradient purification

References

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1.	Louis, N., Fender, P., Barge, A., Kitts, P., and Chroboczek, J. (1994) Cell-binding domain of adenovirus serotype 2 fiber. J Virol 68, 4104–6.

2.	Bergelson, J. M., Cunningham, J. A., Droguett, G., Kurt-Jones, E. A., Krithivas, A., Hong, J. S., et al. (1997). Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5. Science 275, 1320–3.

3.	Santis, G., Legrand, V., Hong, S. S., Davison, E., Kirby, I., Imler, J. L., et al. (1999) Molecular determinants of adenovirus serotype 5 fibre binding to its cellular receptor CAR. J Gen Virol 80(Pt 6), 1519–27.

4.	Bai, M., Harfe, B., and Freimuth, P. (1993) Mutations that alter an Arg-Gly-Asp (RGD) sequence in the adenovirus type 2 penton base protein abolish its cell-rounding activity and delay virus reproduction in flat cells. J Virol 67, 5198–205.

5.	Wickham, T. J., Mathias, P., Cheresh, D. A., and Nemerow, G. R. (1993) Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell 73, 309–19.

6.	Li, E., Brown, S. L., Stupack, D. G., Puente, X. S., Cheresh, D. A., and Nemerow, G. R. (2001) Integrin alpha(v)beta1 is an adenovirus coreceptor. J Virol 75, 5405–9.

7.	Curiel, D. T. (1999) Strategies to adapt adenoviral vectors for targeted delivery. Ann N Y Acad Sci 886, 158–71.

8.	Silman, N. J., and Fooks, A. R. (2000) Biophysical targeting of adenovirus vectors for gene therapy. Curr Opin Mol Ther 2, 524–31.

9.	Mizuguchi, H., and Hayakawa, T. (2004) Targeted adenovirus vectors. Hum Gene Ther 15, 1034–44.

10.	Dmitriev, I., Kashentseva, E., Rogers, B. E., Krasnykh, V., and Curiel, D. T. (2000) Ectodomain of coxsackievirus and adenovirus receptor genetically fused to epidermal growth factor mediates adenovirus targeting to epidermal growth factor receptor-positive cells. J Virol 74, 6875–84.

11.	Zhu, Z. B., Makhija, S. K., Lu, B., Wang, M., Rivera, A. A., Preuss, M., et al. (2004) Transport across a polarized monolayer of Caco-2 cells by transferrin receptor-mediated adenovirus transcytosis. Virology 325, 116–28.

12.	Krasnykh, V. N., Mikheeva, G. V., Douglas, J. T., and Curiel, D. T. (1996) Generation of recombinant adenovirus vectors with modified fibers for altering viral tropism. J Virol 70, 6839–46.

13.	Wickham, T. J., Tzeng, E., Shears, L. L., 2nd, Roelvink, P. W., Li, Y., Lee, G. M., et al. (1997) Increased in vitro and in vivo gene transfer by adenovirus vectors containing chimeric fiber proteins. J Virol 71, 8221–9.

14.	Dmitriev, I., Krasnykh, V., Miller, C. R., Wang, M., Kashentseva, E., Mikheeva, G., et al. (1998) An adenovirus vector with genetically modified fibers demonstrates expanded tropism via utilization of a coxsackievirus and adenovirus receptor-independent cell entry mechanism. J Virol 72, 9706–13.

15.	Wu, H., Seki, T., Dmitriev, I., Uil, T., Kashentseva, E., Han, T., and Curiel, D. T. (2002) Double modification of adenovirus fiber with RGD and polylysine motifs improves coxsackievirus-adenovirus receptor-independent gene transfer efficiency. Hum Gene Ther 13, 1647–53.

16.	Krasnykh, V., Dmitriev, I., Mikheeva, G., Miller, C. R., Belousova, N., and Curiel., D. T. (1998) Characterization of an adenovirus vector containing a heterologous peptide epitope in the HI loop of the fiber knob. J Virol 72, 1844–52.

17.	Belousova, N., Krendelchtchikova, V., Curiel, D. T., and Krasnykh, V. (2002) Modulation of adenovirus vector tropism via incorporation of polypeptide ligands into the fiber protein. J Virol 76, 8621–31.

18.	Xia, H., Anderson, B., Mao, Q., and Davidson, B. L. (2000) Recombinant human adenovirus: targeting to the human transferrin receptor improves gene transfer to brain microcapillary endothelium. J Virol 74, 11359–66.

19.	Wu, H., Dmitriev, I., Kashentseva, E., Seki, T., Wang, M., and Curiel, D. T. (2002) Construction and characterization of adenovirus serotype 5 packaged by serotype 3 hexon. J Virol 76, 12775–82.

20.	Quantum Adeasy $^{TM}$ Vector System Application Manual. (2000) Quantum Biotechnologies, Montreal, Canada.

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