Skip to main content
Log in

In Vivo Gene Transfer by Intravenous Administration of Stable Cationic Lipid/DNA Complex

  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

Abstract

Purpose. A stable cationic lipid/DNA complex has been developed for in vivo gene transfer. The formulation capitalizes on a previously described procedure to obtain stable lipid/DNA complexes for in vitro gene transfer (1).

Methods. Conditions for DNA/lipid complex formation were modified to yield a DNA concentration of 1 mg/ml. Heat stable alkaline phosphatase (AP) under a CMV promoter was used as a reporter gene.

Results. The resulting complex was completely insensitive to serum inactivation. Tail vein injection of a 80 μg DNA into Balb C mice yielded significant levels of reporter enzyme activity in the lung, heart, spleen, muscle, and liver. Less AP activity was observed in the kidney. No AP activity was observed in blood, bone marrow or brain. A titration of the lipid (DOSPA) to DNA-nucleotide ratio showed the optimal molar ratio for in vivo gene transfer to be 1/1. Using this ratio in a dose response study showed approximately 80 μg of DNA/mouse yielded the highest level of gene expression. Using this dose at a 1/1 lipid to DNA nucleotide ratio, the time course for alkaline phosphatase activity was determined. Maximal AP activity was observed 24 hours after injection for all tissues. By day 5, the activity dropped approximately 10 fold for all tissues. By day 7, residual activity was detected in the lung, heart, and muscle. Histology of the lung showed both interstitial and endothelial cells to be transfected. In all other tissues, however, endothelial cells were the only transfected cell type.

Conclusions. These results demonstrate that reformulation of an existing cationic lipid can result in the formation of a stable lipid/DNA complex, which is able to reproducibly transfect lung, heart, spleen, and liver upon intravenous administration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. H. E. J. Hofland, L. Shephard, and S. M. Sullivan. Proc. Natl. Acad. Sci. USA 93:7305–7309 (1996).

    PubMed  Google Scholar 

  2. J. Zabner, A. J. Fasbender, T. Moninger, K. A. Poellinger, and M. J. Welsh. J. Biol. Chem. 270:18997–19007 (1995).

    PubMed  Google Scholar 

  3. S. L. Holmen, M. W. Vanbrocklin, R. R. Eversole, S. R. Stapleton, and L. C. Ginsberg. In Vitro Cell Dev Biol-Animal 31:347–351 (1995).

    Google Scholar 

  4. H. Farhood, N. Serbina, and L. Huang. Biochim. Biophy. Acta 1235:289–295 (1995).

    Google Scholar 

  5. J. H. Felgner, R. Kumar, C. N. Sridhar, C. J. Wheeler, Y. J. Tsai, R. Border, P. Ramsey, M. Martin, and P. L. Felgner. J. Biol. Chem. 269:2550–2561 (1994).

    PubMed  Google Scholar 

  6. O. Boussif, F. Lezoualc'h, M. A. Zanta, M. D. Mergny, D. Scherman, B. Demeneix, and J. P. Behr. Proc. Natl. Acad. Sci. USA 92:7297–7301 (1995).

    PubMed  Google Scholar 

  7. J. Haensler and F. C. Szoka. Bioconjugate Chem. 4:85–93 (1993).

    Google Scholar 

  8. J. C. Perales, T. Ferkol, H. Beegen, O. D. Ratnoff, and R. W. Hanson. Proc. Natl. Acad. Sci. USA 91:4086–4090 (1994).

    PubMed  Google Scholar 

  9. J. Stankovics, A. M. Crane, E. Andrews, C. H. Wu, G. Y. Wu, and F. D. Ledley. Human Gene Ther. 5:1095–1104 (1994).

    Google Scholar 

  10. S. Strydom, P. Van Jaarsveld, E. Van Helden, M. Ariatti, and A. Hawtrey. J. Drug Target. 1:165–174 (1993).

    PubMed  Google Scholar 

  11. K. J. Fisher and J. M. Wilson. Biochem. J. 299:49–58 (1994).

    PubMed  Google Scholar 

  12. S. C. Fields-Berry, A. L. Halliday, and C. L. Cepko. Proc. Natl. Acad. Sci. U. S. A. 89:693–697 (1992).

    PubMed  Google Scholar 

  13. R. J. Samulski, L. S. Chang, and T. Shenk. J. Virol. 63, 3822–3828 (1989).

    PubMed  Google Scholar 

  14. L. Naldini, U. Blomer, P. Gallay, D. Ory, R. Mulligan, F. H. Gage, I. M. Verma, and D. Trono. Science 272:263–267. (1996).

    PubMed  Google Scholar 

  15. B. R. Cullen. Cell 46:973–982 (1986).

    PubMed  Google Scholar 

  16. S. J. Neering, S. F. Hardy, D. Minamoto, S. K. Spratt, and C. T. Jordan. Blood 88:1147–1155 (1996).

    PubMed  Google Scholar 

  17. J. Sambrook, E. F. Fritsch, and T. Maniatis (eds), Molecular Cloning. A laboratory manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 1989, pp. 1.21–1.49.

    Google Scholar 

  18. Y. Liu, D. Liggitt, W. Zhong, G. Tu, K. Gaensler, and R. Debs. J. Biol. Chem. 270:24864–24870 (1995).

    PubMed  Google Scholar 

  19. A. R. Thierry, Y. Lunardi-Iskandar, J. L. Bryant, P. Rabinovich, R. C. Gallo, and L. C. Mahan. Proc. Natl Acad. Sci. 92:9742–9746 (1995).

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hofland, H.E.J., Nagy, D., Liu, JJ. et al. In Vivo Gene Transfer by Intravenous Administration of Stable Cationic Lipid/DNA Complex. Pharm Res 14, 742–749 (1997). https://doi.org/10.1023/A:1012146305040

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1012146305040

Navigation