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  • Nonviral Transfer Technology
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Biodistribution and transgene expression with nonviral cationic vector/DNA complexes in the lungs

Gene Therapy volume 7, pages 1753–1760 (2000)Cite this article

Abstract

Biodistribution of nonviral cationic vector/DNA complexes was studied after systemic or intratracheal administration to the lungs and correlated with transgene expression. Intravenous injection in C57Bl/6 mice gave maximal and significant luciferase expression in the lungs with the cationic polymer PEI 22K/DNA complexes at the highest ratios of positive/negative charges versus DNA alone. While DOTAP/DNA complexes with high charge ratio determined lower but still significant luciferase activity versus uncomplexed DNA, GL-67A and PEI 25K mediated negligible luciferase expression. Labelled PEI 22K and DOTAP complexes were evenly distributed in the alveolar region, where GFP expression was revealed, while PEI 25K and GL-67A complexes were not detected, suggesting a different interaction of these complexes with the plasma membrane of endothelial cells. Following an intratracheal injection, the highest and significant levels of transfection were obtained with slightly positive PEI complexes as compared with DNA alone, whereas cationic lipid-based vectors, DOTAP and GL-67A, gave not significant luciferase activity. Both types of polyplexes gave similar levels of lung luciferase expression by targeting different airway cell populations. PEI 25K complexes determined high levels of GFP in the bronchial cells, confirming confocal data on fluorescent complexes internalization. PEI 22K complexes gave mainly high GFP signal in the distal tract of the bronchial tree, where tagged complexes were recovered. Fluorescent lipid complexes were found in aggregates in the lumen of bronchi totally (DOTAP) or partially (GL-67A) co-localizing with surfactant protein A. Results indicated that cationic polymers could overcome the surfactant barrier which inhibited airway cell transfection mediated by cationic lipids.

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Acknowledgements

This study was supported by a grant for cystic fibrosis from the Italian Ministero della Sanità (L548/93) and the Associazione Lombarda Fibrosi Cistica. G Dina was supported by Giovanni Armenise-Harvard Foundation. We would like to thank SH Cheng for providing us with the GL-67A vector.

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Authors and Affiliations

  1. Institute for Experimental Treatment of Cystic Fibrosis, San Raffaele Scientific Institute, Milano

    A Bragonzi, A Biffi, C Bordignon & M Conese

  2. MIA, DIBIT, San Raffaele Scientific Institute and University of Milano-Bicocca, Milano

    G Dina & A Villa

  3. Epidemiology Unit, San Raffaele Scientific Institute, University of Milano, Italy

    G Calori

  4. Department of Pediatrics, University of Milano, Italy

    B M Assael

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  1. A Bragonzi
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  2. G Dina
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  8. M Conese
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Bragonzi, A., Dina, G., Villa, A. et al. Biodistribution and transgene expression with nonviral cationic vector/DNA complexes in the lungs. Gene Ther 7, 1753–1760 (2000). https://doi.org/10.1038/sj.gt.3301282

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