Abstract
Vectors based on Adenovirus type 5 (Ad5) are among the most common vectors in cancer gene therapy trials to date. However, for increased efficiency and safety, Ad5 should be de-targeted from its native receptors and re-targeted to a tumor antigen. We have described earlier an Ad5 vector genetically re-targeted to the tumor antigen HER2/neu by a dimeric version of the Affibody molecule ZH inserted in the HI-loop of the fiber knob of a coxsackie and adenovirus receptor-binding ablated fiber. This virus showed almost wild-type growth characteristics and infected cells through HER2/neu. Here we generate vectors with double specificity by incorporating two different Affibody molecules, ZH (HER2/neu-binding) and ZT (Taq polymerase-binding), at different positions relative to one another in the HI-loop. Receptor-binding studies together with viral production and gene transfer assays showed that the recombinant fiber with ZT in the first position and ZH in the second position (ZTZH) bound to both its targets, whereas surprisingly, the fiber with ZHZT was devoid of binding to HER2/neu. Hence, it is possible to construct a recombinant adenovirus with dual specificity after evaluating the best position for each ligand in the fiber knob.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Russell WC . Update on adenovirus and its vectors. J Gen Virol 2000; 81: 2573–2604.
Kim M, Zinn KR, Barnett BG, Sumerel LA, Krasnykh V, Curiel DT et al. The therapeutic efficacy of adenoviral vectors for cancer gene therapy is limited by a low level of primary adenovirus receptors on tumour cells. Eur J Cancer 2002; 38: 1917–1926.
Hunt KK, Vorburger SA . Tech.Sight. Gene therapy. Hurdles and hopes for cancer treatment. Science 2002; 297: 415–416.
Barnett BG, Crews CJ, Douglas JT . Targeted adenoviral vectors. Biochim Biophys Acta 2002; 1575: 1–14.
Curiel DT . Strategies to alter the tropism of adenoviral vectors via genetic capsid modifications. In: Curiel DT, Douglas JT (eds). Vector Targeting for Therapeutic Gene Delivery. Wiley-Liss Inc.: Hoboken, 2002, pp 171–200.
Lindholm L, Henning P, Magnusson MK . Novel stratagies in tailoring human adenoviruses into therapeutic cancer gene therapy vectors. Future Medicine 2008; 3: 45–59.
Magnusson MK, Hong SS, Henning P, Boulanger P, Lindholm L . Genetic retargeting of adenovirus vectors: functionality of targeting ligands and their influence on virus viability. J Gene Med 2002; 4: 356–370.
Nilsson B, Moks T, Jansson B, Abrahmsen L, Elmblad A, Holmgren E et al. A synthetic IgG-binding domain based on staphylococcal protein A. Protein Eng 1987; 1: 107–113.
Nord K, Gunneriusson E, Ringdahl J, Stahl S, Uhlen M, Nygren PA . Binding proteins selected from combinatorial libraries of an alpha-helical bacterial receptor domain. Nat Biotechnol 1997; 15: 772–777.
Henning P, Magnusson MK, Gunneriusson E, Hong SS, Boulanger P, Nygren PA et al. Genetic modification of adenovirus 5 tropism by a novel class of ligands based on a three-helix bundle scaffold derived from staphylococcal protein A. Hum Gene Ther 2002; 13: 1427–1439.
Hansson M, Ringdahl J, Robert A, Power U, Goetsch L, Nguyen TN et al. An in vitro selected binding protein (affibody) shows conformation-dependent recognition of the respiratory syncytial virus (RSV) G protein. Immunotechnology 1999; 4: 237–252.
Henning P, Lundgren E, Carlsson M, Frykholm K, Johannisson J, Magnusson MK et al. Adenovirus type 5 fiber knob domain has a critical role in fiber protein synthesis and encapsidation. J Gen Virol 2006; 87: 3151–3160.
Campos SK, Barry MA . Comparison of adenovirus fiber, protein IX, and hexon capsomeres as scaffolds for vector purification and cell targeting. Virology 2006; 349: 453–462.
Kurachi S, Koizumi N, Sakurai F, Kawabata K, Sakurai H, Nakagawa S et al. Characterization of capsid-modified adenovirus vectors containing heterologous peptides in the fiber knob, protein IX, or hexon. Gene Therapy 2007; 14: 266–274.
Magnusson MK, Henning P, Myhre S, Wikman M, Uil TG, Friedman M et al. Adenovirus 5 vector genetically re-targeted by an Affibody molecule with specificity for tumor antigen HER2/neu. Cancer Gene Ther 2007; 14: 468–479.
Codony-Servat J, Albanell J, Lopez-Talavera JC, Arribas J, Baselga J . Cleavage of the HER2 ectodomain is a pervanadate-activable process that is inhibited by the tissue inhibitor of metalloproteases-1 in breast cancer cells. Cancer Res 1999; 59: 1196–1201.
Wikman M, Steffen AC, Gunneriusson E, Adams GP, Carlsson J, Stahl S . Selection and characterization of HER2/neu-binding affibody ligands. Protein Eng Des Sel 2004; 18: 18.
Menard S, Pupa SM, Campiglio M, Tagliabue E . Biologic and therapeutic role of HER2 in cancer. Oncogene 2003; 22: 6570–6578.
Gunneriusson E, Nord K, Uhlen M, Nygren P . Affinity maturation of a Taq DNA polymerase specific affibody by helix shuffling. Protein Eng 1999; 12: 873–878.
Eklund M, Axelsson L, Uhlen M, Nygren PA . Anti-idiotypic protein domains selected from protein A-based affibody libraries. Proteins 2002; 48: 454–462.
Magnusson MK, Hong SS, Boulanger P, Lindholm L . Genetic retargeting of adenovirus: novel strategy employing ‘deknobbing’ of the fiber. J Virol 2001; 75: 7280–7289.
Steffen AC, Wikman M, Tolmachev V, Adams GP, Nilsson FY, Stahl S et al. In vitro characterization of a bivalent anti-HER-2 affibody with potential for radionuclide-based diagnostics. Cancer Biothe Radiopharm 2005; 20: 239–248.
Koizumi N, Mizuguchi H, Sakurai F, Yamaguchi T, Watanabe Y, Hayakawa T . Reduction of natural adenovirus tropism to mouse liver by fiber-shaft exchange in combination with both CAR- and alphav integrin-binding ablation. J Virol 2003; 77: 13062–13072.
Zeng Y, Pinard M, Jaime J, Bourget L, Uyen Le P, O'Connor-McCourt MD et al. A ligand-pseudoreceptor system based on de novo designed peptides for the generation of adenoviral vectors with altered tropism. J Gene Med 2008; 10: 355–367.
Wu H, Seki T, Dmitriev I, Uil T, Kashentseva E, Han T et al. Double modification of adenovirus fiber with RGD and polylysine motifs improves coxsackievirus-adenovirus receptor-independent gene transfer efficiency. Hum Gene Ther 2002; 13: 1647–1653.
Hong JS, Engler JA . Domains required for assembly of adenovirus type 2 fiber trimers. J Virol 1996; 70: 7071–7078.
Friedman M, Orlova A, Johansson E, Eriksson TL, Hoiden-Guthenberg I, Tolmachev V et al. Directed evolution to low nanomolar affinity of a tumor-targeting epidermal growth factor receptor-binding affibody molecule. J Mol Biol 2008; 376: 1388–1402.
Kirby I, Davison E, Beavil AJ, Soh CP, Wickham TJ, Roelvink PW et al. Mutations in the DG loop of adenovirus type 5 fiber knob protein abolish high-affinity binding to its cellular receptor CAR. J Virol 1999; 73: 9508–9514.
O'Reilly DR, Miller LK, Luckow VA . Virus methods. In: Press OU (ed). Baculovirus Expression Vectors. A Laboratory Manual. Oxford University Press Inc: Oxford, England, 1994, pp 124–138.
Mittereder N, March KL, Trapnell BC . Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J Virol 1996; 70: 7498–7509.
Acknowledgements
We are grateful to Affibody AB for providing us with the ZH and ZT Affibody molecules. This study was supported by the European Community (GIANT), contract number: FP6-2003-LIFESCIHEALTH-I 512087 and by grants from the Medical Faculty at the Sahlgrenska Academy: Knut and Alice Wallenbergs stiftelse and Adlerbertska forskningsstiftelsen.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Myhre, S., Henning, P., Friedman, M. et al. Re-targeted adenovirus vectors with dual specificity; binding specificities conferred by two different Affibody molecules in the fiber. Gene Ther 16, 252–261 (2009). https://doi.org/10.1038/gt.2008.160
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/gt.2008.160
Keywords
This article is cited by
-
Development of an adenovirus vector vaccine platform for targeting dendritic cells
Cancer Gene Therapy (2018)
-
Novel viral vectors utilizing intron splice-switching to activate genome rescue, expression and replication in targeted cells
Virology Journal (2011)