Biodistribution and transgene expression with nonviral cationic vector/DNAcomplexes in the lungs

Citation
A. Bragonzi et al., Biodistribution and transgene expression with nonviral cationic vector/DNAcomplexes in the lungs, GENE THER, 7(20), 2000, pp. 1753-1760
Citations number
36
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Molecular Biology & Genetics
Journal title
GENE THERAPY
ISSN journal
0969-7128 → ACNP
Volume
7
Issue
20
Year of publication
2000
Pages
1753 - 1760
Database
ISI
SICI code
0969-7128(200010)7:20<1753:BATEWN>2.0.ZU;2-U
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 C57BI/6 mice gave maximal an d significant luciferase expression in the lungs with the cationic polymer PN 22K/DNA complexes at the highest ratios of positive/negative charges ver sus 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 de tected, suggesting a different interaction of these complexes with the plas ma 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. PN 25K complexes determined high levels of GFP in the bronchial cells, confirming confocal data on flu orescent complexes internalization. PEI 22K complexes gave mainly high GFP signal in the distal tract of the bronchial tree, where tagged complexes we re recovered. Fluorescent lipid complexes were found in aggregates in the l umen of bronchi totally (DOTAP) or partially (GL-67A) co-localizing with su rfactant protein A. Results indicated that cationic polymers could overcome the surfactant barrier which inhibited airway cell transfection mediated b y cationic lipids.