Intravascular and endobronchial DNA delivery to murine lung tissue using a novel, nonviral vector.
Gene transfer to the lung can be achieved via either the airway or the pulmonary vasculature. We evaluated gene transfer and expression by intravascular and endobronchial routes, using DNA complexed with G9 PAMAM dendrimer or naked plasmid DNA. Intravascular tail vein delivery of dendrimer-complexed pCF1CAT plasmid resulted in high levels of transgene expression in the lung at 12 and 24 hr, followed by a second peak of expression 3 to 5 days after administration. After intravenous administration of the complexes, CAT expression was never observed in organs other than the lung. There were only minimal levels of CAT protein expressed in the lung after intravenous administration of naked plasmid DNA. Repeated intravascular doses of the dendrimer-complexed plasmid, administered four times at 4-day intervals, maintained expression at 15-25% of peak concentrations achieved after the initial dose. Endobronchial delivery of naked pCF1CAT plasmid produced significant amounts of CAT protein in the lung. Comparison of intratracheal and intranasal routes resulted in similar expression levels of CAT in the lung and trachea. However, in juxtaposition to vascular delivery, intranasal delivery of dendrimer-complexed plasmid DNA gave lower levels of CAT expression than that observed with naked plasmid DNA. In situ localization of CAT enzymatic activity suggested that vascular administration seemed to achieve expression in the lung parenchyma, mainly within the alveoli, while endobronchial administration primarily targeted bronchial epithelium. Our results show that intravenously administered G9 dendrimer is an effective vector for pulmonary gene transfer and that transgene expression can be prolonged by repeated administration of dendrimer-complexed DNA.
Department of Internal Medicine, University of Michigan Medical School, Ann Arbor 48109, USA.
J F
JF
Kukowska-Latallo
E
E
Raczka
A
A
Quintana
C
C
Chen
M
M
Rymaszewski
J R
JR
Baker
D000281:Administration, Intranasal; D000818:Animals; D001980:Bronchi / Q000378:metabolism* / Q000473:pathology; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D004247:DNA / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005029:Ethylenediamines / Q000737:chemistry; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008168:Lung / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000378:metabolism; D013997:Time Factors; D014132:Trachea / Q000378:metabolism / Q000473:pathology; D019076:Transgenes / Q000235:genetics
D005029:Ethylenediamines; C031234:ethylenediamine; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase