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Molecular Biotechnology

, Volume 54, Issue 2, pp 445–450 | Cite as

An Epidermal Growth Factor Motif from Del1 Protein Increases the Efficiency of In Vivo Gene Transfer with a Non-Viral Vector

  • Atsushi Mamiya
  • Hisataka Kitano
  • Kyoichi Takao
  • Shinichiro Kokubun
  • Masamichi Komiya
  • Chiaki Hidai
Research

Abstract

Increasing the efficiency of gene transfer using non-viral vectors, which have the potential to be safe and economical, would improve upon available options for gene therapy. We previously reported that the third EGF motif of the extracellular matrix protein Del1 (E3) increases the transfection efficiency of non-viral vector methods. Here, we asked if E3 could increase the in vivo transfection efficiency of a polyplex-based approach. To test this, cDNA encoding a heat-stable alkaline phosphatase (AP) was first injected intravenously into mice along with recombinant E3. After 24 h, exogenous AP activity in serum was measured. We found that the introduction of E3 resulted in 50 % more AP activity as compared to the control. We next tested transfection into a tumour explant of SCCKN cells, an oral carcinoma-derived cell line. To do this, a cDNA encoding yellow fluorescent protein was locally injected into a tumour explant, followed by local injection of recombinant E3. Use of E3 increased the number of transfected cells to 2.5 times that of the control. Histochemical staining revealed that E3-induced apoptosis in a tumour explant. The data suggest that E3 might be a useful tool for cancer gene therapy using non-viral vectors.

Keywords

In vivo gene transfer Non-viral vector Del1 EGF motif 

Notes

Acknowledgments

This study was supported by Grant 04-162 from the Japan Science and Technology Agency. We thank Y. Hayashido for kindly providing SCCKN cells and T. Quertermous for kindly providing the Del1 cDNA. English editing was done by FORTE Science Communications.

Conflict of Interest

The authors indicate that they have no competing financial interests.

References

  1. 1.
    Thomas, C. E., Ehrhardt, A., & Kay, M. A. (2003). Progress and problems with the use of viral vectors for gene therapy. Nature Reviews Genetics, 4, 346–358.CrossRefGoogle Scholar
  2. 2.
    Marshall, E. (2000). Gene therapy on trial. Science, 288, 951–957.CrossRefGoogle Scholar
  3. 3.
    Check, E. (2002). Gene therapy: shining hopes dented—but not dashed. Nature, 420, 735.CrossRefGoogle Scholar
  4. 4.
    Magadala, P., & Amiji, M. (2008). Epidermal growth factor receptor-targeted gelatin-based engineered nanocarriers for DNA delivery and transfection in human pancreatic cancer cells. The AAPS Journal, 10, 565–576.CrossRefGoogle Scholar
  5. 5.
    Li, Z., Zhao, R., Wu, X., Sun, Y., Yao, M., Li, J., et al. (2005). Identification and characterization of a novel peptide ligand of epidermal growth factor receptor for targeted delivery of therapeutics. FASEB Journal, 19, 1978–1985.CrossRefGoogle Scholar
  6. 6.
    Saul, J. M., Annapragada, A., Natarajan, J. V., & Bellamkonda, R. V. (2003). Controlled targeting of liposomal doxorubicin via the folate receptor in vitro. Journal of Controlled Release, 92, 49–67.CrossRefGoogle Scholar
  7. 7.
    Zhao, X. B., & Lee, R. J. (2004). Tumor-selective targeted delivery of genes and antisense oligodeoxyribonucleotides via the folate receptor. Advanced Drug Delivery Reviews, 56, 1193–1204.CrossRefGoogle Scholar
  8. 8.
    Oba, M., Fukushima, S., Kanayama, N., Aoyagi, K., Nishiyama, N., Koyama, H., et al. (2007). Cyclic RGD peptide-conjugated polyplex micelles as a targetable gene delivery system directed to cells possessing alphavbeta3 and alphavbeta5 integrins. Bioconjugate Chemistry, 18, 1415–1423.CrossRefGoogle Scholar
  9. 9.
    Kunath, K., Merdan, T., Hegener, O., Haberlein, H., & Kissel, T. (2003). Integrin targeting using RGD-PEI conjugates for in vitro gene transfer. Journal of Gene Medicine, 5, 588–599.CrossRefGoogle Scholar
  10. 10.
    Kitano, H., Hidai, C., Kawana, M., & Kokubun, S. (2008). An epidermal growth factor-like repeat of Del1 protein increases the efficiency of gene transfer in vitro. Molecular Biotechnology, 39, 179–185.CrossRefGoogle Scholar
  11. 11.
    Hidai, C., Zupancic, T., Penta, K., Mikhail, A., Kawana, M., Quertermous, E. E., et al. (1998). Cloning and characterization of developmental endothelial locus-1: An embryonic endothelial cell protein that binds the alphavbeta3 integrin receptor. Genes & Development, 12, 21–33.CrossRefGoogle Scholar
  12. 12.
    Hidai, C., Kawana, M., Habu, K., Kazama, H., Kawase, Y., Iwata, T., et al. (2005). Overexpression of the Del1 gene causes dendritic branching in the mouse mesentery. The Anatomical Record Part A Discoveries in Molecular, Cellular, and Evolutionary Biology, 287, 1165–1175.CrossRefGoogle Scholar
  13. 13.
    Kitano, H., Kokubun, S., & Hidai, C. (2010). The extracellular matrix protein Del1 induces apoptosis via its epidermal growth factor motif. Biochemical and Biophysical Research Communications, 393, 757–761.CrossRefGoogle Scholar
  14. 14.
    Urade, M., Ogura, T., Mima, T., & Matsuya, T. (1992). Establishment of human squamous carcinoma cell lines highly and minimally sensitive to bleomycin and analysis of factors involved in the sensitivity. Cancer, 69, 2589–2597.CrossRefGoogle Scholar
  15. 15.
    Berger, J., Hauber, J., Hauber, R., Geiger, R., & Cullen, B. R. (1988). Secreted placental alkaline phosphatase: A powerful new quantitative indicator of gene expression in eukaryotic cells. Gene, 66, 1–10.CrossRefGoogle Scholar
  16. 16.
    Lowe, M. E. (1992). Site-specific mutations in the COOH-terminus of placental alkaline phosphatase: A single amino acid change converts a phosphatidylinositol-glycan-anchored protein to a secreted protein. Journal of Cell Biology, 116, 799–807.CrossRefGoogle Scholar
  17. 17.
    Bowers, W. J., Breakefield, X. O., & Sena-Esteves, M. (2011). Genetic therapy for the nervous system. Human Molecular Genetics, 20, R28–R41.CrossRefGoogle Scholar
  18. 18.
    Kuwahara, H., Nishina, K., Yoshida, K., Nishina, T., Yamamoto, M., Saito, Y., et al. (2011). Efficient in vivo delivery of siRNA into brain capillary endothelial cells along with endogenous lipoprotein. Molecular Therapy, 19, 2213–2221.CrossRefGoogle Scholar
  19. 19.
    Litzinger, D. C., Brown, J. M., Wala, I., Kaufman, S. A., Van, G. Y., Farrell, C. L., et al. (1996). Fate of cationic liposomes and their complex with oligonucleotide in vivo. Biochimica et Biophysica Acta, 1281, 139–149.CrossRefGoogle Scholar
  20. 20.
    Hidai, C., Kawana, M., Kitano, H., & Kokubun, S. (2007). Discoidin domain of Del1 protein contributes to its deposition in the extracellular matrix. Cell and Tissue Research, 330, 83–95.CrossRefGoogle Scholar
  21. 21.
    Hidai, C., Kitano, H., & Kokubun, S. (2009). The Del1 deposition domain can immobilize 3alpha-hydroxysteroid dehydrogenase in the extracellular matrix without interfering with enzymatic activity. Bioprocess and Biosystems Engineering, 32, 569–573.CrossRefGoogle Scholar
  22. 22.
    Kitano, H., Mamiya, A., & Hidai, C. (2011). Improvement of FasL gene therapy in vitro by fusing the FasL to Del1 protein domains. In Y. You (Ed.), Targets in Gene Therapy. Croatia: In Tech Rijeka.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Atsushi Mamiya
    • 1
  • Hisataka Kitano
    • 2
  • Kyoichi Takao
    • 1
  • Shinichiro Kokubun
    • 1
  • Masamichi Komiya
    • 2
  • Chiaki Hidai
    • 1
  1. 1.Division of Physiology, Department of Biomedical SciencesNihon University School of MedicineTokyoJapan
  2. 2.Division of Dental SurgeryNihon University School of MedicineTokyoJapan

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