Rapid crossing of the pulmonary endothelial barrier by polyethylenimine/DNA complexes.
Intravenous administration could become a delivery route of choice for prophylactic and curative gene therapies on condition that genes cross the capillary barrier and reach target tissues without being degraded. We investigated the kinetics and process of transgene delivery through mouse lung capillaries following DNA complexation with linear polyethylenimine (L-PEI) and intravenous injection. Using digoxin-labeled DNA we followed the cellular localization of DNA at different times after injection and correlated these findings with cell markers and transgene expression. At 2 h after injection some DNA was still localized on the interior of the capillary lumen, but other complexes had already crossed the barrier and resulted in gene expression. At 24 h after injection most labeled DNA was localised in pulmonary cells, as was transgene expression. Only rarely was transgene expression found in endothelial cells, suggesting that the complexes cross the capillary barrier rapidly. Levels of caspase-1-like activity did not increase following transfection implying that L-PEI/DNA complexes are transported across cellular barriers by a non-damaging, physiological process, without causing inflammation. The high levels of expression of different transgenes in pneumocytes indicates that transport of L-PEI/DNA complexes through the endothelial barrier does not affect their transfection capacity. These findings open up new possibilities for gene delivery and its application to the lung.
Goula
D
D
Laboratoire de Physiologie Générale et Comparée, UMR CNRS 8572, Muséum National d'Histoire Naturelle, Paris, France.
Becker
N
N
Lemkine
G F
GF
Normandie
P
P
Rodrigues
J
J
Mantero
S
S
Levi
G
G
Demeneix
B A
BA
D000818:Animals; D002196:Capillaries; D004247:DNA / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000493:pharmacokinetics*; D007275:Injections, Intravenous; D008099:Liver / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D008168:Lung / Q000098:blood supply / Q000378:metabolism*; D051379:Mice; D011094:Polyethyleneimine / Q000378:metabolism*; D013997:Time Factors; D019076:Transgenes
D011094:Polyethyleneimine; D004247:DNA; D008156:Luciferases