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Fiber-modified adenovirus vectors containing the TAT peptide derived from HIV-1 in the fiber knob have efficient gene transfer activity

Gene Therapy volume 14, pages 1160–1165 (2007)Cite this article

Abstract

The interaction between viral capsid proteins and specific molecules exposed on the plasma membrane of the cells is involved in the viral tropism. A human adenovirus (Ad) belonging to subgroups A, C, D, E and F infects cells via the interaction between the fiber knob and the primary receptor, the coxsackievirus and adenovirus receptor (CAR). Conventional human adenovirus type 5 (hAd5) vectors show efficient transduction in CAR-positive cells; in contrast, hAd5 vector application is limited by poor transduction into cells lacking CAR expression. In the present study, to broaden the tropism of hAd5 vectors, we generated hAd5 vectors containing the TAT peptide, which is a protein transduction domain derived from human immunodeficiency virus, in the HI loop of the fiber knob (Ad-TAT(HI)-L2) or the C-terminus of the fiber knob (Ad-TAT(C)-L2). In CAR-negative adherent cells, Ad-TAT(HI)-L2 and Ad-TAT(C)-L2 showed approximately 50- to 500-fold higher gene expression than the conventional hAd5 vector (Ad-L2). Ad-TAT(HI)-L2 was also more efficient than Ad-L2 in blood cell lines and in two types of primary cultured human vascular smooth muscle cells, which are almost refractory to Ad-L2. Furthermore, Ad-TAT(HI)-L2 was more efficient than other types of fiber-modified Ad vectors, which harbor an RGD (Arg-Gly-Asp) or a poly-lysine (KKKKKKK;K7) peptide in the HI loop or the C-terminus of the fiber knob, respectively. Ad-TAT(HI)-L2 efficiently transduced the organs in levels and patterns that were roughly similar to those of Ad-L2 after being systemically injected into mice. To the best of our knowledge, this study is the first report showing that hAd5 vectors containing the TAT peptide in the fiber knob could efficiently transduce cells independently of CAR. These Ad vectors should be useful for gene functional analysis and gene therapy.

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References

  1. Kovesdi I, Brough DE, Bruder JT, Wickham TJ . Adenoviral vectors for gene transfer. Curr Opin Biotechnol 1997; 8: 583–589.

    Article  CAS  PubMed  Google Scholar 

  2. Benihoud K, Yeh P, Perricaudet M . Adenovirus vectors for gene delivery. Curr Opin Biotechnol 1999; 10: 440–447.

    Article  CAS  PubMed  Google Scholar 

  3. Wickham TJ . Targeting adenovirus. Gene Ther 2000; 7: 110–114.

    Article  CAS  PubMed  Google Scholar 

  4. Mizuguchi H, Hayakawa T . Targeted adenovirus vectors. Hum Gene Ther 2004; 15: 1034–1044.

    Article  CAS  PubMed  Google Scholar 

  5. van Tuyn J, Atsma DE, Winter EM, van der Velde-van Dijke I, Pijnappels DA, Bax NA et al. Epicardial cells of human adults can undergo an epithelial-to-mesenchymal transition and obtain characteristics of smooth muscle cells in vitro. Stem Cells 2007; 25: 271–278.

    Article  CAS  PubMed  Google Scholar 

  6. Wickham TJ, Tzeng E, Shears II LL, Roelvink PW, Li Y, Lee GM et al. Increased in vitro and in vivo gene transfer by adenovirus vectors containing chimeric fiber proteins. J Virol 1997; 71: 8221–8229.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Dmitriev I, Krasnykh V, Miller CR, Wang M, Kashentseva E, Mikheeva G et al. An adenovirus vector with genetically modified fibers demonstrates expanded tropism via utilization of a coxsackievirus and adenovirus receptor-independent cell entry mechanism. J Virol 1998; 72: 9706–9713.

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Vigne E, Mahfouz I, Dedieu JF, Brie A, Perricaudet M, Yeh P . RGD inclusion in the hexon monomer provides adenovirus type 5-based vectors with a fiber knob-independent pathway for infection. J Virol 1999; 73: 5156–5161.

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Koizumi N, Mizuguchi H, Hosono T, Ishii-Watabe A, Uchida E, Utoguchi N et al. Efficient gene transfer by fiber-mutant adenoviral vectors containing RGD peptide. Biochim Biophys Acta 2001; 1568: 13–20.

    Article  CAS  PubMed  Google Scholar 

  10. Mizuguchi H, Koizumi N, Hosono T, Utoguchi N, Watanabe Y, Kay MA et al. A simplified system for constructing recombinant adenoviral vectors containing heterologous peptides in the HI loop of their fiber knob. Gene Therapy 2001; 8: 730–735.

    Article  CAS  PubMed  Google Scholar 

  11. Dmitriev IP, Kashentseva EA, Curiel DT . Engineering of adenovirus vectors containing heterologous peptide sequences in the C terminus of capsid protein IX. J Virol 2002; 76: 6893–6899.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Koizumi N, Mizuguchi H, Utoguchi N, Watanabe Y, Hayakawa T . Generation of fiber-modified adenovirus vectors containing heterologous peptides in both the HI loop and C terminus of the fiber knob. J Gene Med 2003; 5: 267–276.

    Article  CAS  PubMed  Google Scholar 

  13. Vellinga J, Rabelink MJ, Cramer SJ, van den Wollenberg DJ, Van der Meulen H, Leppard KN et al. Spacers increase the accessibility of peptide ligands linked to the carboxyl terminus of adenovirus minor capsid protein IX. J Virol 2004; 78: 3470–3479.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. 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.

    Article  CAS  PubMed  Google Scholar 

  15. Gupta B, Levchenko TS, Torchilin VP . Intracellular delivery of large molecules and small particles by cell-penetrating proteins and peptides. Adv Drug Deliv Rev 2005; 57: 637–651.

    Article  CAS  PubMed  Google Scholar 

  16. Mizuguchi H, Kay MA . Efficient construction of a recombinant adenovirus vector by an improved in vitro ligation method. Hum Gene Ther 1998; 9: 2577–2583.

    Article  CAS  PubMed  Google Scholar 

  17. Mizuguchi H, Kay MA . A simple method for constructing E1- and E1/E4-deleted recombinant adenoviral vectors. Hum Gene Ther 1999; 10: 2013–2017.

    Article  CAS  PubMed  Google Scholar 

  18. Arap W, Pasqualini R, Ruoslahti E . Cancer treatment by targeted drug delivery to tumor vasculature in a mouse model. Science 1998; 279: 377–380.

    Article  CAS  PubMed  Google Scholar 

  19. Maizel Jr JV, White DO, Scharff MD . The polypeptides of adenovirus. I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. Virology 1968; 36: 115–125.

    Article  CAS  PubMed  Google Scholar 

  20. Shen H, Mai JC, Qiu L, Cao S, Robbins PD, Cheng T . Evaluation of peptide-mediated transduction in human CD34+ cells. Hum Gene Ther 2004; 15: 415–419.

    Article  CAS  PubMed  Google Scholar 

  21. Kawabata K, Sakurai F, Yamaguchi T, Hayakawa T, Mizuguchi H . Efficient gene transfer into mouse embryonic stem cells with adenovirus vectors. Mol Ther 2005; 12: 547–554.

    Article  CAS  PubMed  Google Scholar 

  22. Sakurai F, Kawabata K, Yamaguchi T, Hayakawa T, Mizuguchi H . Optimization of adenovirus serotype 35 vectors for efficient transduction in human hematopoietic progenitors: comparison of promoter activities. Gene Therapy 2005; 12: 1424–1433.

    Article  CAS  PubMed  Google Scholar 

  23. Tyagi M, Rusnati M, Presta M, Giacca M . Internalization of HIV-1 tat requires cell surface heparan sulfate proteoglycans. J Biol Chem 2001; 276: 3254–3261.

    Article  CAS  PubMed  Google Scholar 

  24. Console S, Marty C, Garcia-Echeverria C, Schwendener R, Ballmer-Hofer K . Antennapedia and HIV transactivator of transcription (TAT) ‘protein transduction domains’ promote endocytosis of high molecular weight cargo upon binding to cell surface glycosaminoglycans. J Biol Chem 2003; 278: 35109–35114.

    Article  CAS  PubMed  Google Scholar 

  25. Weeks BS, Desai K, Loewenstein PM, Klotman ME, Klotman PE, Green M et al. Identification of a novel cell attachment domain in the HIV-1 Tat protein and its 90-kDa cell surface binding protein. J Biol Chem 1993; 268: 5279–5284.

    CAS  PubMed  Google Scholar 

  26. Vogel BE, Lee SJ, Hildebrand A, Craig W, Pierschbacher MD, Wong-Staal F et al. A novel integrin specificity exemplified by binding of the alpha v beta 5 integrin to the basic domain of the HIV Tat protein and vitronectin. J Cell Biol 1993; 121: 461–468.

    Article  CAS  PubMed  Google Scholar 

  27. Schwarze SR, Ho A, Vocero-Akbani A, Dowdy SF . In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 1999; 285: 1569–1572.

    Article  CAS  PubMed  Google Scholar 

  28. Gratton JP, Yu J, Griffith JW, Babbitt RW, Scotland RS, Hickey R et al. Cell-permeable peptides improve cellular uptake and therapeutic gene delivery of replication-deficient viruses in cells and in vivo. Nat Med 2003; 9: 357–362.

    Article  CAS  PubMed  Google Scholar 

  29. Kuhnel F, Schulte B, Wirth T, Woller N, Schafers S, Zender L et al. Protein transduction domains fused to virus receptors improve cellular virus uptake and enhance oncolysis by tumor-specific replicating vectors. J Virol 2004; 78: 13743–13754.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Xu ZL, Mizuguchi H, Ishii-Watabe A, Uchida E, Mayumi T, Hayakawa T . Optimization of transcriptional regulatory elements for constructing plasmid vectors. Gene 2001; 272: 149–156.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Yuriko Teramura for technical assistance. This work was supported by grants from the Ministry of Health, Labor, and Welfare of Japan.

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Authors and Affiliations

  1. Laboratory of Gene Transfer and Regulation, National Institute of Biomedical Innovation, Osaka, Japan

    S Kurachi, K Tashiro, F Sakurai, H Sakurai, K Kawabata & H Mizuguchi

  2. Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan

    S Kurachi, K Tashiro, H Sakurai, S Nakagawa & H Mizuguchi

  3. Department of Biopharmaceutical Sciences, Kobe Gakuin University, Hyogo, Japan

    K Yayama & H Okamoto

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  1. S Kurachi
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  2. K Tashiro
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Correspondence to H Mizuguchi.

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Kurachi, S., Tashiro, K., Sakurai, F. et al. Fiber-modified adenovirus vectors containing the TAT peptide derived from HIV-1 in the fiber knob have efficient gene transfer activity. Gene Ther 14, 1160–1165 (2007). https://doi.org/10.1038/sj.gt.3302969

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