Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Factors influencing the efficiency of cationic liposome-mediated intravenous gene delivery

Nature Biotechnology volume 15, pages 167–173 (1997)Cite this article

Abstract

We have characterized the relationships between the design of cationic liposomes as a gene transfer vehicle, their resulting biodistribution and processing in animals, and the level and sites of gene expression they produce. By redesigning conventional cationic liposomes, incorporating cholesterol (chol) as the neutral lipid and preparing them as multilamellar vesicles (MLV), we increased the efficiency of cationic liposome:DNA complex (CLDC)-mediated gene delivery. Expression of the luciferase gene increased up to 1,740-fold and of the human granulocyte-colony stimulating factor (hG-CSF) gene up to 569-fold due to prolonged circulation time of injected CLDC, and increased uptake and retention in tissues. The level of gene expression per microgram of DMA taken up per tissue was 1,000-fold higher in lung than in liver, indicating that in addition to issues of delivery and retention of injected DNA, tissue-specific host factors also play a central role in determining the efficiency of expression. Vascular endothelial cells, monocytes, and macrophages are the cell types most commonly transfected by intravenous injection of CLDC.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Herz, J. and Gerard, R.D. 1993. Adenovirus-mediated transfer of low density lipoprotein receptor gene acutely accelerates cholesterol clearance in normal mice. Proc. Natl. Acad. Sci. USA 90: 2812–2816.

    Article  CAS  Google Scholar 

  2. Kozarsky, K.F., McKinley, D.R., Austin, L.L., Raper, S.E., Stratford-Perricaudet, L.D. and Wilson, J.M. 1994. In vivo correction of low density lipoprotein receptor deficiency in the Watanabe heritable hyperlipidemic rabbit with recombinant adenoviruses. J. Biol. Chem. 269: 13965–13702.

    Google Scholar 

  3. Yang, Y., Nunes, F.A., Berencsi, K., Furth, E.E., Gonczol, E. and Wilson, J.M. 1994. Cellular immunity to viral antigens limits E1 -deleted adenoviruses for gene therapy. Proc. Natl. Acad. Sci. USA 91: 4407–4411.

    Article  CAS  Google Scholar 

  4. Knowles, M.R., Hohneker, K.W., Zhou, Z., Olsen, J.C., Noah, T.L., Hu, P.C., Leigh, M.W., Engelhardt, J.F., Edwards, L.J., Jones, K.R. and Boucher, R. 1995. A controlled study of adenoviral-vector-mediated gene transfer in the nasal epithelium of patients with cystic fibrosis. New Engl. J. Med. 333: 823–831.

    Article  CAS  Google Scholar 

  5. Stewart, M.J., Plautz, G.E., Del Buono, L., Yang, Z.Y., Xu, L., Gao, X., Huang, L., Nabel, E.G. and Nabel, G.J. 1992. Gene transfer in vivo with DNA-liposome complexes: safety and acute toxicity in mice. Hum. Gene Ther. 3: 267–275.

    Article  CAS  Google Scholar 

  6. Zhu, N., Liggitt, D., Liu, Y. and Debs, R. 1993. Systemic gene expression after intravenous DNA delivery into adult mice. Science 261: 209–211.

    Article  CAS  Google Scholar 

  7. Canonico, A.E., Plitman, J.D., Conary, J.T., Meyrick, B.O. and Brigham, K.L. 1994. No lung toxicity after repeated aerosol or intravenous delivery of plasmid-cationic liposome complexes. J. Appl. Physiol. 77: 415–419.

    Article  CAS  Google Scholar 

  8. Thierry, A.R., Iskandar, Y.L., Bryant, J.C., Rabinovich, P., Gallo, R.C. and Marian, L.C. 1995. Systemic gene therapy: biodistribution and long-term expression of a transgene in mice. Proc. Natl. Acad. Sci. USA 92: 9742–9746.

    Article  CAS  Google Scholar 

  9. Liu, Y., Liggitt, D., Tu, G., Zhong, W., Gaensler, K., and Debs, R. 1995. Cationic lipo-some-mediated intravenous gene delivery in mice. J. Biol. Chem. 27: 24864–24870.

    Article  Google Scholar 

  10. Strauss, W.M., Dawsman, J., Beard, C., Johnson, C., Lawrence, J.B. and Jaenisch, R. 1993. Germ line transmission of a yeast artificial chromosome spanning the murine alpha 1 (I) collagen locus. Science 259: 1904–1906.

    Article  CAS  Google Scholar 

  11. Solodin, I., Brown, C., Bruno, M., Chow, C., Jangm, E-H., Debs, R., and Heath, T. 1995. High effciency in vivo gene delivery with a novel series of amphilic imada-zolinium compounds. Biochemistry 34: 13537–13544.

    Article  CAS  Google Scholar 

  12. Feigner, P., Gadek, T., Holm, M., Roman, R., Chan, H.W., Wenz, M., Northrop, T.P., Ringold, G.M. and Danielson, M. 1987. Lipofection: a highly efficient, lipid-mediated DNA transfection procedure. Proc. Natl. Acad. Sci. USA 84: 7413–7420.

    Article  Google Scholar 

  13. Leventis, R. and Silvius, J. 1990. Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles. Biochim. Biophys. Acta 1023: 124–130.

    Article  CAS  Google Scholar 

  14. Rose, J., Buonocore, L. and Whitt, M.A. 1991. A new cationic liposome reagent mediating nearly quantitative transfection of animal cells. BioTechniques 10: 520–526.

    CAS  PubMed  Google Scholar 

  15. Feigner, J.H., Kumar, R., Sridhar, C.N., Wheeler, C.J., Tsal, Y.J., Border, R., Ramsay, P., Martin, M. and Feigner, P.L. 1994. Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations. J. Biol. Chem. 269: 2550–2561.

    Google Scholar 

  16. Brigham, K., Meyrick, B., Christman, B., Magnuson, M., King, G. and Berry, L.C. 1989. In vivo transfection of murine lungs with a functioning prokaryotic gene using a liposome vehicle. Am. J. Med. Sci. 298: 278–286.

    Article  CAS  Google Scholar 

  17. Hyde, S.C., Gill, D.R., Higgins, C.F., Trezise, A.E., MacVinish, L.J., Cuthbert, A.W., Ratcliff, R., Evans, M.J. and Colledge, W.H. 1993. Correction of the ion transport defect in cystic fibrosis transgenic mice by gene therapy. Nature 362: 250–253.

    Article  CAS  Google Scholar 

  18. Canonico, A.E., Conary, J.T., Meyrick, B.O. and Brigham, K.L. 1994. Aerosol and intravenous transfection of human alpha 1 -antitrypsin gene to lungs of rabbits. Am. J. Resp. Cell. Mol. Biol. 10: 24–29.

    Article  CAS  Google Scholar 

  19. Nabel, E.G., Plautz, G. and Nabel, G.J. 1990. Site-specific gene expression in vivo by direct gene transfer into the arterial wall. Science 249: 1285–1288.

    Article  CAS  Google Scholar 

  20. Alton, E.W., Middleton, P.G., Caplen, N.J., Smith, S.N., Steel, D.M., Munkonge, P.M., Jeffery, P.K., Stevenson, B.J., McLachlan, G., Dorin, J.R. and Porteous, D.J. 1993. Non-invasive liposome-mediated gene delivery can correct the ion transport defect in cystic fibrosis mutant mice. Nature Genet. 5: 135–142.

    Article  CAS  Google Scholar 

  21. Logan, J.J., Bebok, Z., Walker, L.C., Peng, S., Feigner, P.L., Siegal, G.P., Frizzell, R.A., Dong, J., Howard, M., Matalon, S., Lindsey, J.R., DuVall, M. and Sorscher, E.J. 1995. Cationic lipids for reporter gene and CFTR transfer to rat pulmonary epithelium. Gene Therapy 2: 38–49.

    CAS  PubMed  Google Scholar 

  22. Son, K. and Huang, L. 1994. Exposure of human ovarian carcinoma to cisplatin transiently sensitizes the tumor cells for liposome-mediated gene transfer. Proc. Natl. Acad. Sci. USA 91: 12669–12672.

    Article  CAS  Google Scholar 

  23. Nabel, G.J., Nabel, E.G., Yang, Z.-Y., Fox, B.A., Plautz, G.E., Gao, X., Huang, L., Shu, S., Gordon, D. and Chang, A.E. 1993. Direct gene transfer with DNA-liposome complexes in melanoma: expression, biologic activity, and lack of toxicity in humans. Proc. Natl. Acad. Sci. USA 90: 1130–1138.

    Google Scholar 

  24. Caplen, N.J., Alton, E.W., Middleton, P.G., Dorin, J.R., Stevenson, B.J., Gao, X., Durham, S.R., Jeffery, P.K., Hodson, M.E., Coutelle, C., Huang, L., Porteous, D.J., Willaimson, R. and Geddes, D.M. 1995. Liposome-mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis. Nature Med. 1: 39–46.

    Article  CAS  Google Scholar 

  25. Conary, J.T., Parker, R.E., Christman, B.W., Faulks, R.D., King, G.A., Meyrick, B.O. and Brigham, K.L. 1994. Protection of rabbit lungs from endotoxin injury by in vivo hyperex-pnession of the prostaglandin G/H synthase gene. J. Clin. Invest. 93: 1834–1840.

    Article  CAS  Google Scholar 

  26. Lesoon-Wood, L.A., Kim, W.H., Kleinman, H.K., Weintraub, B.D. and Mixson, A. 1995. Systemic gene therapy with p53 reduces growth and metastases of a malignant human breast cancer in nude mice. Hum. Gene Ther. 6: 395–405.

    Article  CAS  Google Scholar 

  27. Fasbender, A.J., Zabner, J. and Welsh, M.J. 1995. Optimization of cationic lipid-mediated gene transfer to airway epithelia. Amer. J. Physiol. 269(Pt 1): L4551

    Article  CAS  Google Scholar 

  28. Sorscher, E.J., Logan, J.J., Frizzell, R.A., Lyrene, R.K., Bebok, Z., Dong, J.Y., Duvall, M.D., Feigner, P.L., Matalon, S., Walker, L. and Wiatrak, B.J. 1994. Gene therapy for cystic fibrosis using cationic liposome mediated gene transfer: a phase I trial of safety and efficacy in the nasal airway. Hum. Gene Ther. 5: 1259–1277.

    Article  CAS  Google Scholar 

  29. Wheeler, C.J., Feigner, P.L., Tsai, Y.J., Marshall, J., Sukhu, L., Doh, G., Hartikka, J., Nietupski, J., Manthorpe, M., Nichols, M., Rewe, M., Liang, X., Norman, J., Smith, A. and Cheng, S.H. 1996. A novel cationic lipid greatly enhances plasmid DNA delivery and expression in mouse lung. Proc. Natl. Acad. Sci. USA 93: 11454–11459.

    Article  CAS  Google Scholar 

  30. Jiao, S., Acsadi, G., Jani, A., Feigner, P.L. and Wolff, J.A. 1992. Persistence of plasmid DNA and expression in rat brain cells in vivo. Expt. Neurol. 115: 400–413.

    Article  CAS  Google Scholar 

  31. Bennett, M.J., Nantz, M.H., Balasubramaniam, R.P., Gruenert, D.C. and Malone, R.W. 1995. Cholesterol enhances cationic liposome-mediated DNA transfection of human respiratory epithelial cells. Bioscience Reports 15: 47–53.

    Article  CAS  Google Scholar 

  32. Weis, J., Fine, S.M., David, C., Savarirayan, S. and Sanes, J.R. 1991. Integration site-dependent expression of a transgene reveals specialized features of cells associated with neuromuscular junctions. J. Cell. Biol. 113: 1385–1397.

    Article  CAS  Google Scholar 

  33. Sternberg, B., Sorgi, F.L. and Huang, L. 1994. New structures in complex formation between DNA and cationic liposomes visualized by freeze-fracture electron microscopy. FEBS Lett. 356: 361–366.

    Article  CAS  Google Scholar 

  34. Gershon, H., Ghirlando, R., Guttmsan, S.B. and Minsky, A. 1993. Mode of formation and structural features of DNA-cationic liposome complexes used for transfection. Biochemistry 32: 71434–7151.

    Article  Google Scholar 

  35. Zabner, J., Fasbender, A.J., Moninger, T., Poellinger, K.A. and Welsh, M.J. 1995. Cellular and molecular barriers to gene transfer by a cationic lipid. J. Biol. Chem. 270: 18997–19007.

    Article  CAS  Google Scholar 

  36. Senior, J. and Gregoriadis, G. 1982. Stability of small unilamellar liposomes in serum and clearance from the circulation: the effect of the phospholipid and cholesterol components. Life Sciences 30: 2123–2136.

    Article  CAS  Google Scholar 

  37. Mayhew, E., Rustum, Y.M., Szoka, F.C. and Papahadjopoulos, D. 1979. Role of cholesterol in enhancing the antitumor activity of cytosine arabinoside entrapped in liposomes. Cancer Treat. Reports 63: 1923–1928.

    CAS  Google Scholar 

  38. Hofland, H.E., Shephard, L. and Sullivan, S.M. 1996. Formation of stable cationic lipid/DNA complexes for gene transfer. Proc. Natl. Acad. Sci. USA 93: 7305–7309.

    Article  CAS  Google Scholar 

  39. Petros, W.P. 1992. Pharmacokinetics and administration of colony-stimulating factors. Pharmacotherapy 12: 32S–38S

    CAS  PubMed  Google Scholar 

  40. Pauksen, K., Elfman, L., Ulfgren, A. and Venge, P. 1994. Serum levels of granu-locyte-colony stimulating factor (G-CSF) in bacterial and viral infections, and in atypical pneumonia. Brit. J. Haem. 88: 256–260.

    Article  CAS  Google Scholar 

  41. Kawakami, M., Tsutsumi, H., Kumakawa, T., Abe, H., Hirai, M., Kurosawa, S., Mori, M. and Fukushima, M. 1990. Levels of serum granulocyte colony-stimulating factor in patients with infections. Blood 76: 1962–1964.

    CAS  PubMed  Google Scholar 

  42. Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular cloning, A laboratory manual, Second edition. Ford, N., Nolan, C., and Ferguson, M. (eds.). Cold Spring Harbor Laboratory Press.

    Google Scholar 

  43. Bligh, E.G. and Dyer, W.J. 1959. A rapid method of total lipid extraction and purification. Can, J. Bioch. Physiol. 37: 911–917.

    Article  CAS  Google Scholar 

  44. Debs, R.J., Straubinger, R.M., Brunette, E.N., Lin, J.M., Lin, E.J., Montgomery, A.B., Friend, D.S. and Papahadjopoulos, D.P. 1987. Selective enhancement of pentamidine uptake in the lung by aerosolization and delivery in liposomes. Amer. Rev. Respir. Dis. 135: 731–737.

    CAS  Google Scholar 

  45. Johnston, R.F., Pickett, S.L. and Barker, D.L. 1990. Autoradiography using storage phosphor technology. Electrophoresis 11: 355–360.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Robert J. Debs: (e-mail: karin@itsa.ucsf.edu).

  2. Yong Liu and Leslie C. Mounkes: Authors contributed equally.

Authors and Affiliations

  1. California Pacific Medical Research Institute, 2330 Clay St., San Francisco, CA, 94115

    Yong Liu, Leslie C. Mounkes & Robert J. Debs

  2. Department of Comparative Medicine, University of Washington, Seattle, WA, 98195

    H. Denny Liggitt

  3. School of Pharmacy, University of Wisconsin-Madison, WI, 53706

    Carolyn S. Brown, Igor Solodin & Timothy D. Heath

Authors
  1. Yong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Leslie C. Mounkes
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H. Denny Liggitt
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Carolyn S. Brown
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Igor Solodin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Timothy D. Heath
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Robert J. Debs
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, Y., Mounkes, L., Liggitt, H. et al. Factors influencing the efficiency of cationic liposome-mediated intravenous gene delivery. Nat Biotechnol 15, 167–173 (1997). https://doi.org/10.1038/nbt0297-167

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0297-167

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing