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.

  • Paper
  • Published:

Comparison between cationic polymers and lipids in mediating systemic gene delivery to the lungs

Gene Therapy volume 6, pages 1995–2004 (1999)Cite this article

Abstract

Airway inflammation frequently found in congenital and acquired lung diseases may interfere with gene delivery by direct administration through either instillation or aerosol. Systemic delivery by the intravenous administration represents an alternative route of delivery that might bypass this barrier. A nonviral approach for transfecting various airway-derived cell lines in vitro showed that cationic polymers (PEI 22K and 25K) and lipids (DOTAP, GL-67/DOPE) are able to transfect with high efficiency the reporter genes firefly luciferase and E. coli lacZ. Notably, two properties predicted that cationic vectors would be useful for a systemic gene delivery approach to the lung: (1) transfection was not inhibited or increased when cells were incubated with cationic lipids or polymers in the presence of serum; and (2) cationic vectors protected plasmid DNA from DNase degradation. A single injection of DNA complexed to the cationic polymer PEI 22K into the tail vein of adult mice efficiently transfected primarily the lungs and to a lesser extent, heart, spleen, kidney and liver. The other vectors mediated lower to undetectable levels of luciferase expression in the lungs, with DOTAP > GL67/DOPE > PEI 25K > DOTMA/DOPE. A double injection protocol with a 15-min interval between the two doses of DOTAP/DNA complexes was investigated and showed a relevant role of the first injection in transfecting the lungs. A two log increase in luciferase expression was obtained either when the two doses were comprised of luciferase plasmid or when an irrelevant plasmid was used in the first injection. The double injection of luciferase/PEI 22K complexes determined higher transgene levels than a single dose, but a clear difference using an irrelevant plasmid as first dose was not observed. Using lacZ as a reporter gene, it was shown that only cells in the alveolar region, including type II penumocytes, stained positively for the transgene product.

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
Figure 6

Similar content being viewed by others

References

  1. Grubb B et al. Inefficient gene transfer by adenovirus vector to cystic fibrosis airway epithelia of mice and humans Nature 1994 371: 802–806

    Article  CAS  Google Scholar 

  2. Korst RJ, Bewig B, Crystal RG . In vitro and in vivo transfer and expression of human surfactant SP-A- and SP-B-associated protein cDNAs mediated by replication-deficient, recombinant adenoviral vectors Hum Gene Ther 1995 6: 277–287

    Article  CAS  Google Scholar 

  3. Epperly M et al. Prevention of late effects of irradiation lung damage by manganese superoxide dismutase gene therapy Gene Therapy 1998 5: 196–208

    Article  CAS  Google Scholar 

  4. Ferkol T et al. Transfer of the human alpha1-antitrypsin gene into pulmonary macrophages in vivo Am J Respir Cell Mol Biol 1998 18: 591–601

    Article  CAS  Google Scholar 

  5. Fasbender AJ, Zabner J, Welsh MJ . Optimization of cationic lipid-mediated gene transfer to airway epithelial Am J Physiol 1995 13: L45–L51

    Google Scholar 

  6. McLachlan G et al. Evaluation in vitro and in vivo of cationic liposome-expression construct complexes for cystic fibrosis gene therapy Gene Therapy 1995 2: 614–622

    CAS  PubMed  Google Scholar 

  7. Biffi A et al. Restoration of bacterial killing activity of human respiratory cystic fibrosis cells through cationic vector-mediated CFTR gene transfer Hum Gene Ther 1999 10: 1923–1930

    Article  CAS  Google Scholar 

  8. Stribling R et al. Aerosol gene delivery in vivo Proc Natl Acad Sci USA 1992 89: 11277–11281

    Article  CAS  Google Scholar 

  9. Alton EWFW et al. Non-invasive liposome-mediated gene delivery can correct the ion transport defect in cystic fibrosis mutant mice Nat Genet 1993 5: 135–142

    Article  CAS  Google Scholar 

  10. Hyde LJ et al. Correction of the ion transport defect in cystic fibrosis transgenic mice by gene therapy Nature 1993 362: 250–255

    Article  CAS  Google Scholar 

  11. Eastman SJ et al. Optimization of formulations and conditions for the aerosol delivery of functional cationic lipid:DNA complexes Hum Gene Ther 1997 8: 313–322

    Article  CAS  Google Scholar 

  12. Zhu N et al. Systemic gene expression after intravenous DNA delivery into adult mice Science 1993 261: 209–211

    Article  CAS  Google Scholar 

  13. Liu Y et al. Cationic liposome-mediated intravenous gene delivery J Biol Chem 1995 270: 24864–24870

    Article  CAS  Google Scholar 

  14. Thierry AR et al. Systemic gene therapy: biodistribution and long-term expression of a transgene in mice Proc Natl Acad Sci USA 1995 92: 9742–9746

    Article  CAS  Google Scholar 

  15. Thierry AR et al. Characterization of liposome-mediated gene delivery: expression, stability and pharmacokinetics of plasmid DNA Gene Therapy 1997 4: 226–237

    Article  CAS  Google Scholar 

  16. Li S, Huang L . In vivo gene transfer via intravenous administration of cationic lipid–protamine–DNA (LPD) complexes Gene Therapy 1997 4: 891–900

    Article  CAS  Google Scholar 

  17. Liu Y et al. Factors influencing the efficiency of cationic liposome-mediated intravenous gene delivery Nat Biotechnol 1997 15: 167–173

    Article  CAS  Google Scholar 

  18. Griesenbach U et al. Comparison between intratracheal and intravenous administration of liposome–DNA complexes for cystic fibrosis lung gene therapy Gene Therapy 1998 5: 181–188

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  20. Boussif O, Zanta MA, Behr J-P . Optimized galenics improve in vitro gene transfer with cationic molecules up to 1000-fold Gene Therapy 1996 3: 1074–1080

    CAS  Google Scholar 

  21. Boletta A et al. Nonviral gene delivery to the rat kidney with polyethylenimine Hum Gene Ther 1997 8: 1243–1251

    Article  CAS  Google Scholar 

  22. 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 

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

    Article  CAS  Google Scholar 

  24. Felgner PL et al. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure Proc Natl Acad Sci USA 1987 84: 7413–7417

    Article  CAS  Google Scholar 

  25. Song YK et al. Characterization of cationic liposome-mediated gene transfer in vivo by intravenous administration Hum Gene Ther 1997 8: 1585–1594

    Article  CAS  Google Scholar 

  26. Abdallah B et al. A powerful nonviral vector for in vivo gene transfer into the adult mammalian brain: polyethylenimine Hum Gene Ther 1996 7: 1947–1954

    Article  CAS  Google Scholar 

  27. Dunlop DD et al. Nanoscopic structure of DNA condensed for gene delivery Nucleic Acids Res 1997 25: 3095–3101

    Article  Google Scholar 

  28. Ogris M et al. The size of DNA/transferrin–PEI complexes is an important factor for gene expression in cultured cells Gene Therapy 1998 5: 1425–1433

    Article  CAS  Google Scholar 

  29. Goula D et al. Size, diffusibility and transfection performance of linear PEI/DNA complexes in the mouse central nervous system Gene Therapy 1998 5: 712–717

    Article  CAS  Google Scholar 

  30. Schwartz DA et al. CpG motifs in bacterial DNA cause inflammation in the lower respiratory tract J Clin Invest 1997 100: 68–73

    Article  CAS  Google Scholar 

  31. Plank C et al. Activation of the complement system by synthetic DNA complexes: a potential barrier for intravenous gene delivery Hum Gene Ther 1996 7: 1437–1446

    Article  CAS  Google Scholar 

  32. Bragonzi A et al. Glomerular filtration is required for transfection of proximal tubular cells in the rat kidney following injection of DNA complexes in to the renal artery (submitted, revised)

  33. Lee ER et al. Detailed analysis of structures and formulations of cationic lipids for efficient gene transfer to the lung Hum Gene Ther 1996 7: 1701–1717

    Article  CAS  Google Scholar 

  34. Song YK, Liu F, Liu D . Enhanced gene expression in mouse lung by prolonging the retention time of intravenously injected plasmid DNA Gene Therapy 1998 5: 1531–1537

    Article  CAS  Google Scholar 

  35. Song YK, Liu D . Free liposomes enhance the transfection activity of DNA/lipid complexes in vivo by intravenous administration Biochim Biophys Acta 1998 1372: 141–150

    Article  CAS  Google Scholar 

  36. McCray PB et al. Alveolar macrophages inhibit retrovirus-mediated gene transfer to airway epithelia Hum Gene Ther 1997 8: 1087–1093

    Article  CAS  Google Scholar 

  37. McCray P, Wohlford-Lenane C, Snyder J . Localisation of cystic fibrosis transmembrane conductance regulator mRNA in human fetal lung tissue by in situ hybridization J Clin Invest 1992 90: 619–625

    Article  CAS  Google Scholar 

  38. Sambrook J, Fritsch EF, Maniatis T . Molecular Cloning. A Laboratory Manual Cold Spring Harbor Laboratory Press: Cold Spring Harbor 1989

    Google Scholar 

  39. Behr J-P et al. Efficient gene transfer into mammalian primary endocrine cells with lipopolyamine-coated DNA Proc Natl Acad Sci USA 1989 86: 6982–6986

    Article  CAS  Google Scholar 

  40. Alcorn JL et al. Primary cell culture of human type II pneumocytes: maintenance of a differentiated phenotype and transfection with recombinant adenoviruses Am J Respir Cell Mol Biol 1997 17: 672–682

    Article  CAS  Google Scholar 

  41. Springer TA et al. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody Eur J Immunol 1979 9: 301–306

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by a grant for Cystic Fibrosis from the Italian Ministero della Sanità (L 548/93) and the Associazione Lombarda Fibrosi Cistica. We would like to thank Dr D Gruenert for the 56FHT80- and CFNPE90- cell lines, Dr L Monaco for the pCLuc and pCMVβn plasmids, Dr J-P Behr for the DOGS cationic lipid, Dr J Alcorn for the anti-SP-A polyclonal antibody, and Dr I Simon for the anti-CD11b monoclonal antibody.

Author information

Authors and Affiliations

  1. Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Milano, Italy

    A Bragonzi, A Boletta, A Biffi, A Muggia, C Bordignon & M Conese

  2. Milano and Department of Pediatrics, Centro di Riferimento Fibrosi Cistica, University of Milano, Italy

    G Sersale & B M Assael

  3. Genzyme Corporation, Framingham, MA, USA

    S H Cheng

Authors
  1. A Bragonzi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A Boletta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Biffi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Muggia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G Sersale
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S H Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. C Bordignon
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. B M Assael
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. M Conese
    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

Bragonzi, A., Boletta, A., Biffi, A. et al. Comparison between cationic polymers and lipids in mediating systemic gene delivery to the lungs. Gene Ther 6, 1995–2004 (1999). https://doi.org/10.1038/sj.gt.3301039

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301039

Keywords

This article is cited by

Search

Advanced search

Quick links