In vivo gene delivery to the lung using polyethylenimine and fractured polyamidoamine dendrimers.
BACKGROUND
Gene transfer into the airways could be of importance for the treatment of chronic lung diseases such as cystic fibrosis. In the past few years several attempts have been made to effectively deliver DNA to the lung using different viral and non-viral vector systems. Viral vectors and cationic lipids have been tested intensively but the properties of cationic polymers such as polyethylenimine (PEI) 25 kDa and fractured polyamidoamine dendrimers to deliver DNA to the airways have not been studied. Surfactant preparations have been shown to influence pulmonary adenoviral and naked plasmid DNA mediated gene transfer in vivo. We investigated the gene delivery efficiency of branched PEI 25 kDa and fractured dendrimers to the murine lung in vivo and also examined the effect of surfactant on PEI 25 kDa mediated gene transfer to the lung.


METHODS
Cationic polymer/DNA complexes were prepared in 25 mM HEPES buffer (pH = 7.4) or double distilled water and administered to the lungs of BALB/c mice via cannula intubation. The trachea, left and right lung, heart, liver and esophagus were examined for luciferase activity. Inflammation was assessed by performing standard histology.


RESULTS
PEI/DNA complexes showed a high level of luciferase gene expression in the lung. Complexes formed in double distilled water exhibited higher gene expression than complexes formed in 25 mM HEPES buffer (pH 7.4). The optimal N/P ratio was found to be N/P = 10 in double distilled water. Luciferase activity was only detected in the lung and decreased rapidly in a time-dependent manner. The addition of a natural surfactant preparation, Alveofact, slightly reduced gene transfer of branched PEI 25 kDa. Luciferase gene expression obtained by using fractured dendrimers was very low.


CONCLUSIONS
The present study demonstrates that PEI 25 kDa, but not polyamidoamine dendrimers, effectively mediates transient gene transfer to the murine lung after intratracheal intubation. In conclusion, branched PEI 25 kDa was found to be an effective vector for pulmonary gene delivery in vivo, being superior to fractured dendrimers.
Rudolph
C
C
Department of Pediatrics, University of Munich, Germany.
Lausier
J
J
Naundorf
S
S
Müller
R H
RH
Rosenecker
J
J
D000818:Animals; D004274:DNA, Recombinant / Q000494:pharmacology*; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D007442:Intubation, Intratracheal; D008055:Lipids; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010743:Phospholipids; D011073:Polyamines; D011094:Polyethyleneimine; D011663:Pulmonary Surfactants; D013501:Surface-Active Agents
D004274:DNA, Recombinant; D008055:Lipids; D010743:Phospholipids; D011073:Polyamines; D011663:Pulmonary Surfactants; C078272:SF-RI 1, bovine surfactant preparation; D013501:Surface-Active Agents; D011094:Polyethyleneimine; D008156:Luciferases