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:

A comparison of linear and branched polyethylenimine (PEI) with DCChol/DOPE liposomes for gene delivery to epithelial cells in vitro and in vivo

Gene Therapy volume 10, pages 1654–1662 (2003)Cite this article

Abstract

Polyethylenimine (PEI), a polycation with high ionic charge density, has recently been used as a gene therapy delivery agent. We have defined the optimal conditions for PEI-based transfection of airway epithelial cells in vitro and in vivo and used these conditions to restore Cl− channel activity in a CF mouse model. Three forms of PEI, a linear 22 kDa (ExGen 500) form and branched 25 or 50 kDa forms were evaluated. All forms of PEI significantly increased luciferase reporter gene expression compared to the liposome DCChol/DOPE in a human bronchial epithelial cell line (16HBE) irrespective of the extent of cell confluency. With subconfluent cells, gene expression was around 1000-, 200- and 25-fold higher than liposomes using linear 22, 25 and 50 kDa PEI, respectively. The transfection efficiency was reduced in confluent and polarized epithelial cells but linear 22 kDa PEI showed the smallest decrease and gave 8000-fold better transfection in polarized cells compared to liposomes. A comparison of linear 22 or 25 kDa PEI with DCChol/DOPE for airway delivery in vivo via intranasal instillation was also performed. Linear 22 kDa PEI gave significantly better luciferase reporter gene expression of 350-fold in the lung, 180-fold in the nose and 85-fold in the trachea compared to liposome. In contrast, the 25 kDa form of PEI was no better than DCChol/DOPE. Repeat dosing with linear 22 kDa PEI failed to give reporter gene delivery comparable to the initial dose. To establish that PEI can be used to deliver a physiologically relevent gene in vivo, we used it to restore Cl− secretion by CFTR gene delivery in the airways of a CF mouse model.

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Riordan JR et al. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 1989; 245: 1066–1073.

    Article  CAS  Google Scholar 

  2. Gill DR et al. A placebo-controlled study of liposome-mediated gene transfer to the nasal epithelium of patients with cystic fibrosis. Gene Therapy 1997; 4: 199–209.

    Article  CAS  Google Scholar 

  3. Hyde SC et al. Repeat administration of DNA/liposomes to the nasal epithelium of patients with cystic fibrosis. Gene Therapy 2000; 7: 1156–1165.

    Article  CAS  Google Scholar 

  4. Alton EW et al. Cationic lipid-mediated CFTR gene transfer to the lungs and nose of patients with cystic fibrosis: a double-blind placebo-controlled trial. Lancet 1999; 353: 947–954.

    Article  CAS  Google Scholar 

  5. Knowles MR et al. A double-blind, placebo controlled, dose ranging study to evaluate the safety and biological efficacy of the lipid–DNA complex GR213487B in the nasal epithelium of adult patients with cystic fibrosis. Hum Gene Ther 1998; 9: 249–269.

    Article  CAS  Google Scholar 

  6. Zabner J et al. Comparison of DNA–lipid complexes and DNA alone for gene transfer to cystic fibrosis airway epithelia in vivo. J Clin Invest 1997; 100: 1529–1537.

    Article  CAS  Google Scholar 

  7. Porteus DJ et al. Evidence for safety and efficacy of DOTAP cationic liposome mediated CFTR gene transfer to the nasal epithelium of patients with cystic fibrosis. Gene Therapy 1997; 4: 210–218.

    Article  Google Scholar 

  8. Boussif O et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethyleneimine. Proc Natl Acad Sci USA 1995; 92: 7297–7301.

    Article  CAS  Google Scholar 

  9. Goula D et al. Polyethylenimine-based intravenous delivery of transgenes to mouse lung. Gene Therapy 1998; 5: 1291–1295.

    Article  CAS  Google Scholar 

  10. Godbey WT, Wu KK, Mikos AG . Tracking the intracellular path of poly(ethylenimine)/DNA complexes for gene delivery. Proc Natl Acad Sci USA 1999; 96: 5177–5181.

    Article  CAS  Google Scholar 

  11. Remy JS et al. Taking lipospermines and polyethylenimine from in vitro to in vivo. In: Kabanov AV, Felgner PL, Seymour LW (eds). Self-assembling Complexes for Gene Delivery: From Laboratory to Clinical trial. John Wiley and Sons Ltd.: New York.

  12. Ferrari S et al. ExGen 500 is an efficient vector for gene delivery to lung epithelial cells in vitro and in vivo. Gene Therapy 1997; 4: 1100–1106.

    Article  CAS  Google Scholar 

  13. Ferrari S et al. Polyethylenimine shows properties of interest for cystic fibrosis gene therapy. Biochim Biophys Acta 1999; 1447: 219–225.

    Article  CAS  Google Scholar 

  14. Bragonzi A et al. Comparison between cationic polymers and lipids in mediating systemic gene delivery to the lungs. Gene Therapy 1999; 6: 1995–2004.

    Article  CAS  Google Scholar 

  15. Goula D et al. Rapid crossing of the pulmonary endothelial barrier by polyethlenimine/DNA complexes. Gene Therapy 2000; 7: 499–504.

    Article  CAS  Google Scholar 

  16. Bragonzi A et al. Biodistribution and transgene expression with nonviral cationic vector/DNA complexes in the lungs. Gene Therapy 2000; 7: 1753–1760.

    Article  CAS  Google Scholar 

  17. Zou SM, Erbacher P, Remy JS, Behr JP . Systemic linear polyethylenimine (L-PEI)-mediated gene delivery in the mouse. J Gene Med 2000; 2: 128–134.

    Article  CAS  Google Scholar 

  18. Chollet P, Favrot MC, Hurbin A, Coll JL . Side effects of a systemic injection of linear polyethylenimine–DNA complexes. J Gene Med 2002; 4: 84–91.

    Article  Google Scholar 

  19. Densmore CL et al. Aerosol delivery of robust polyethyleneimine–DNA complexes for gene therapy and genetic immunization. Mol Ther 2000; 1: 180–188.

    Article  CAS  Google Scholar 

  20. Gautam A, Densmore CL, Xu B, Waldrep JC . Enhanced gene expression in mouse lung after PEI–DNA aerosol delivery. Mol Ther 2000; 2: 63–70.

    Article  CAS  Google Scholar 

  21. Cozens AL et al. CFTR expression and chloride secretion in polarized immortal human bronchial epithelial cells. Am J Respir Cell Mol Biol 1994; 10: 38–47.

    Article  CAS  Google Scholar 

  22. Cozens AL et al. A transformed human epithelial cell line that retains tight junctions post crisis. In Vitro Cell Dev Biol 1992; 28A: 735–744.

    Article  CAS  Google Scholar 

  23. Krieg AM et al. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature 1995; 374: 546–549.

    Article  CAS  Google Scholar 

  24. Ratcliff R et al. Production of a severe cystic fibrosis mutation in mice by gene targeting. Nat Genet 1993; 4: 35–41.

    Article  CAS  Google Scholar 

  25. Goddard CA, Evans MJ, Colledge WH . Genistein activates CFTR-mediated Cl− secretion in the murine trachea and colon. Am J Physiol 2000; 279C: 383-C392.

    Google Scholar 

  26. Brunner S et al. Overcoming the nuclear barrier: cell cycle independent nonviral gene transfer with linear polyethylenimine or electroporation. Mol Ther 2002; 5: 80–86.

    Article  CAS  Google Scholar 

  27. Mortimer I et al. Cationic lipid-mediated transfection of cells in culture requires mitotic activity. Gene Therapy 1999; 6: 403–411.

    Article  CAS  Google Scholar 

  28. Li S et al. Effect of immune response on gene transfer to the lung via systemic administration of cationic lipidic vectors. Am J Physiol 1999; 276L: 796-804.

    Google Scholar 

  29. Uduehi AN et al. Effects of linear polyethylenimine and polyethylenimine/DNA on lung function after airway instillation to rat lungs. Mol Therapy 2001; 4: 52–57.

    Article  CAS  Google Scholar 

  30. Hartikka J et al. An improved plasmid DNA expression vector for direct injection into skeletal muscle. Hum Gene Therapy 1996; 7: 1205–1217.

    Article  CAS  Google Scholar 

  31. MacVinish LJ et al. Chloride secretion in the trachea of null cystic fibrosis mice: the effects of transfection with pTrial10-CFTR2. J Phys 1997; 499: 677–687.

    CAS  Google Scholar 

  32. Gautam A et al. Transgene expression in mouse airway epithelium by aerosol gene therapy with PEI–DNA complexes. Mol Ther 2001; 3: 551–556.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the CF Trust (UK). WHC is supported by the Ford Physiology Fund.

Author information

Authors and Affiliations

  1. Department of Physiology, University of Cambridge, Cambridge, UK

    J W Wiseman, C A Goddard, D McLelland & W H Colledge

Authors
  1. J W Wiseman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C A Goddard
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D McLelland
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. W H Colledge
    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

Wiseman, J., Goddard, C., McLelland, D. et al. A comparison of linear and branched polyethylenimine (PEI) with DCChol/DOPE liposomes for gene delivery to epithelial cells in vitro and in vivo. Gene Ther 10, 1654–1662 (2003). https://doi.org/10.1038/sj.gt.3302050

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3302050

Keywords

This article is cited by

Search

Advanced search

Quick links