Rapid crossing of the pulmonary endothelial barrier by polyethylenimine/DNA complexes

Goula, Daniel; Becker, Nathalie; Lemkine, Gregory F.; Normandie, Priscilla; Rodrigues, Joana S.; Mantero, Stefano; Levi, Giovanni; Demeneix, Barbara

doi:10.1038/sj.gt.3301113

Cited by 137 publications

(101 citation statements)

References 22 publications

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“…36,37 PEI complexes have been shown to enter pulmonary epithelial cells, as well as other lung tissue cells after intravenous injection. 27 The efficiency of PEImediated transfection in vitro can be enhanced by improving the packing of PEI molecules in the complex, 38 likely through better buffering in the lysosome that is essential for effective transfection. 35 These PEI effects, however, can result in considerable toxicity with large quantities of the DNA and polycation.…”

Section: Discussionmentioning

confidence: 99%

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Gene delivery to the lung using protein/polyethylenimine/plasmid complexes

et al. 2002

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Section: Discussionmentioning

confidence: 99%

“…14,26,27 The rate of injection for HSA/PEI/DNA preparations was assessed using a range of injection times between 3 s (the minimum time necessary for a single smooth i.v. push injection of 200 l via tail vein) and 5-10 l pulse injections distributed over 30 s to 3 min.…”

Section: Influence Of Injection Rate Incubation Time and Injection Vmentioning

confidence: 99%

Gene delivery to the lung using protein/polyethylenimine/plasmid complexes

et al. 2002

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“…13 Although DOTAP and PEI 22K labeled complexes were biodistributed in a similar fashion, the transgene levels in the lungs were higher for PEI 22K. It may be possible that highly compacted PEI 22K complexes favorably cross the endothelium, as was shown for pulmonary endothelium in mice 26 and endothelial glomerular barrier in the rat kidney. 25 Moreover, greater diffusibility of these complexes related to their small size and stability was also demonstrated in the mouse central nervous system.…”

Section: Figure 5 Immunohistochemical Detection Of Gfp Transgene Exprmentioning

confidence: 92%

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

Bragonzi¹,

et al. 2000

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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

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“…The higher distribution to the lung in the early phase might be due to the entrapment of PEI/DNA complexes in the pulmonary capillaries, or the result of ionic association with the large surface area of the lung endothelia. Since PEI/DNA complexes can be transported across the pulmonary endothelial barrier within a few hours, 15 it is likely that plasmid DNA in the lung at 24 h might reside in pulmonary cells.…”

Section: The Density Of Each Band Was Measured Using a Gel-doc Image mentioning

confidence: 99%

Prolonged organ retention and safety of plasmid DNA administered in polyethylenimine complexes

Kim

Yoon

et al. 2001

Gene Ther

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Polyethylenimine (PEI) Recently, polyethylenimine (PEI) has drawn attention as a versatile, inexpensive, and useful nonviral transfection vector. 1 A variety of cell types such as monocytes, 2 dendritic cells, 3 myoblast cells, 4 and hepatocytes 5 were studied as target cells for PEI-mediated gene transfection. In vivo, PEI was shown to be an efficient transfection vector in several organs such as the kidney, liver, and lung. [6][7][8] However, these studies are mostly focused on the expression of the PEI/DNA limited to target organs in vivo or specific cell types in vitro. Although the knowledge of biodistribution fates of plasmid DNA may reveal the feasibility of the cellular level targeting in vivo, there is little understanding of the overall and quantitative organ distribution of plasmid DNA after administration in PEI complexes. Moreover, the safety of PEI/DNA complexes after repeated dosing has not been well documented, despite the potential requirement for repeated administration due to the episomal property of plasmids.In this study, we tested the distribution fate and safety of plasmid DNA after administering DNA in PEI complexes. Quantitative and competitive polymerase chain reaction (PCR) was employed for the assay of plasmid DNA in the biological samples. Here, we report that in comparison to naked DNA, the organ distribution was higher and remarkably prolonged after administration of plasmid DNA in PEI complexes, and that once a week dosing of PEI/DNA complexes over 3 weeks did not alter the histology of major organs such as the liver, lung, and kidney.PEI/DNA complexes with PEI nitrogen to DNA phosphate ratio (N/P ratio) of 10:1 were prepared in 5% glucose using 25 kDa PEI (Aldrich, St Quentin, France). The N/P ratio of 10:1 was chosen since it was shown to efficiently transfect cells in vivo. 9 The levels of plasmid DNA in biological samples were determined using competitive and quantitative PCR. Quantitative PCR is based on co-amplification of the sample template together with various amounts of internal standard (IS) competitor sharing with the target the primer recognition sites, but differing in size. The competitor was constructed to share the same sense and antisense primer used for target amplification. 10 As an example, Figure 1 shows the gel bands and internal standard curves obtained after running competitive PCR of DNA extracts from the liver 6 h after injection of naked DNA or PEI/DNA complexes. The IS for quantitative PCR was the 188-bp deleted mutant of pCMV␤. The construction of the competitor and PCR conditions were described previously. 10 The PCR products were 449 bp and 261 bp for the target and the IS, respectively. The band densities of the target gene PCR product diminished as the amounts of IS increased (Figure 1a and b). The quantitation of gel band density by densitometry generated internal standard curves of high linearity (Figure 1c). The curves also show that we could detect as low as fg-level plasmids by quantitative PCR. The amounts of plasmid DNA in the samples we...

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Rapid crossing of the pulmonary endothelial barrier by polyethylenimine/DNA complexes

Cited by 137 publications

References 22 publications

Gene delivery to the lung using protein/polyethylenimine/plasmid complexes

Gene delivery to the lung using protein/polyethylenimine/plasmid complexes

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

Prolonged organ retention and safety of plasmid DNA administered in polyethylenimine complexes

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