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Session V – DNA Transfer Vehicles

Visualization of the transgene distribution according to the administration route allows prediction of the transfection efficacy and validation of the results obtained

Abstract

Gene transfer to the lung can be achieved via a systemic, that targets the endothelium, or local, that targets the epithelium, delivery route. In the present study, we followed the distribution of a plasmid after transfection using some of our phosphonolipids, which have previously shown their efficiency in transfecting mouse lungs. The plasmid was radiolabeled and varying combinations of plasmid/phosphonolipid were administered by intravenous injection, or by endotracheal spray. The distribution of radioactive labeling was observed over a time course using a γ-camera. These images were then correlated with the results for luciferase expression levels in the lungs. In each case, lungs were well targeted. However, whereas an intravenous injection reaches all of the lung immediately, progressive diffusion occurs when the plasmid/phosphonolipid is administered via an aerosol. Elimination of the radioactivity associated with plasmid occurs via the urinary tract after intravenous injections, and via the feces using the aerosol delivery approach. The radioactivity detected in the lungs correlated strongly with transgene expression. Thus, such an imaging technique is a powerful strategy to predict the formulation that will generate the best transfection efficiency. This study reveals that scintigraphic imaging permits both validation of the administration method and the results obtained for each animal, thereby reducing the statistical variability of in vivo experiments.

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References

  1. Bellon G et al. Aerosol administration of a recombinant adenovirus expressing CFTR to cystic fibrosis patients: a phase I clinical trial Hum Gene Ther 1997 8: 15–25

    Article  CAS  Google Scholar 

  2. Perricone MA et al. Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. II. Transfection efficiency in airway epithelium Hum Gene Ther 2001 12: 1383–1394

    Article  CAS  Google Scholar 

  3. Boucher RC et al. Gene therapy for cystic fibrosis using E1-deleted adenovirus: a phase I trial in the nasal cavity. The University of North Carolina at Chapel Hill Hum Gene Ther 1994 5: 615–639

    Article  CAS  Google Scholar 

  4. Joseph PM et al. Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. I. Methods, safety, and clinical implications Hum Gene Ther 2001 12: 1369–1382

    Article  CAS  Google Scholar 

  5. Floch V et al. Systemic administration of cationic lipids/DNA complexes and the relationship between formulation and lung transfection efficiency Biochim Biophys Acta 2000 1464: 95–103

    Article  CAS  Google Scholar 

  6. Barron LG, Gagne L, Szoka FC Jr . Lipoplex-mediated gene delivery to the lung occurs within 60 minutes of intravenous administration Hum Gene Ther 1999 10: 1683–1694

    Article  CAS  Google Scholar 

  7. Guillaume-Gable C et al. Cationic phosphonolipids as nonviral gene transfer agents in the lungs of mice Hum Gene Ther 1998 9: 2309–2319

    Article  CAS  Google Scholar 

  8. Alton EWFW 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 

  9. Chadwick SL et al. Safety of a single aerosol administration of escalating doses of the cationic lipid GL-67/DOPE/DMPE-PEG 5000 formulation to the lungs of normal volunteers Gene Therapy 1997 4: 937–942

    Article  CAS  Google Scholar 

  10. Ishiwata H et al. Characteristics and biodistribution of cationic liposomes and their DNA complexes J Control Rel 2000 65: 139–148

    Article  Google Scholar 

  11. 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 

  12. Thierry AR et al. Systemic gene therapy: biodistribution and long-term expression of a transgene in mice Delépine P et al. Phosphonolipids: a class of nonviral vectors rapidly eliminated and efficient in mice lung-directed gene transfer. Hum Gene Ther (submitted)

  13. Uyechi LS et al. Mechanism of lipoplex gene delivery in mouse lung: binding and internalization of fluorescent lipid and DNA components Gene Therapy 2001 8: 828–836

    Article  CAS  Google Scholar 

  14. Osaka G et al. Pharmacokinetics, tissue distribution, and expression efficiency of plasmid [33P]DNA following intravenous administration of DNA/cationic lipid complexes in mice: use of a novel radionuclide approach J Pharm Sci 1996 85: 612–618

    Article  CAS  Google Scholar 

  15. Lerondel S et al. Radioisotopic imaging allows optimization of adenovirus lung deposition for cystic fibrosis gene therapy Hum Gene Ther 2001 12: 1–11

    Article  CAS  Google Scholar 

  16. Sun S et al. Quantitative imaging of gene induction in living animals Gene Therapy 2001 8: 1572–1579

    Article  CAS  Google Scholar 

  17. Delépine P et al. Cationic phosphonolipids as nonviral vectors: in vitro and in vivo applications J Pharm Sci 2000 89: 629–638

    Article  Google Scholar 

  18. Guillaume C et al. Characterization and optimization of aerosolized cationic lipid-DNA complexes Biochem Biophys Res Com 2001 286: 464–471

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the French associations ‘Vaincre la Mucoviscidose’ and ‘Association pour la recherche sur le cancer’ (ARC) and by the conseil general de Bretagne.

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Delepine, P., Montier, T., Guillaume, C. et al. Visualization of the transgene distribution according to the administration route allows prediction of the transfection efficacy and validation of the results obtained. Gene Ther 9, 736–739 (2002). https://doi.org/10.1038/sj.gt.3301742

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