title: In vivo gene delivery to the pulmonary circulation in rats: transgene distribution and vascular inflammatory response.
abstract: Although gene delivery to the pulmonary circulation has both experimental and therapeutic potential, the delivery methods, distribution of transgene, and subsequent inflammatory response have been poorly characterized to date. To address these issues, we utilized a 0.76-mm OD (outside diameter) end hole catheter inserted into the internal jugular vein of adult Sprague-Dawley rats, directing the tip into a pulmonary capillary wedge position. We then compared infusion of polycationic lipid:DNA complexes to replication-defective adenovirus with respect to magnitude and distribution of transgene expression using either chloramphenicol acetyltransferase (CAT) or human placental alkaline phosphatase (hpAP) reporter genes. Both lipid:DNA and adenovirus resulted in detectable transgene expression, though maximum lung CAT activity using lipid (gamma AP-DLRIE/DOPE) was approximately 2% of maximum activity using adenovirus (Ad-CAT). Further characterization of expression after transfection with 10(8) pfu (plaque forming units) of Ad-CAT demonstrated persistence of transgene for at least 14 days (lung CAT activity 27% of maximum). Alkaline phosphatase staining demonstrated that both large and small pulmonary arteries as well as the alveolar wall expressed transgene. Although little inflammatory response was detected in conduit arteries, a predominantly mononuclear cell infiltrate surrounded small pulmonary arteries as well as the alveolar spaces in transfected areas of lung. We conclude that percutaneous catheter-mediated gene delivery to the pulmonary circulation in rats using non-viral and viral vectors is feasible. Although an inflammatory response to first generation replication-defective adenovirus was detected, it appeared to be largely restricted to the distal pulmonary circulation and airspace. This technique should prove useful for investigations requiring overexpression of novel genes in the pulmonary artery wall, and could ultimately be used to develop gene-based therapies for pulmonary vascular diseases.
authors.affiliation: Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado Health Sciences Center, Denver 80262, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Rodman
authors.forename: H
authors.initials: H
authors.lastname: San
authors.forename: R
authors.initials: R
authors.lastname: Simari
authors.forename: D
authors.initials: D
authors.lastname: Stephan
authors.forename: F
authors.initials: F
authors.lastname: Tanner
authors.forename: Z
authors.initials: Z
authors.lastname: Yang
authors.forename: G J
authors.initials: GJ
authors.lastname: Nabel
authors.forename: E G
authors.initials: EG
authors.lastname: Nabel
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002404:Catheterization; D002795:Choline O-Acetyltransferase / Q000235:genetics; D004247:DNA / Q000737:chemistry; D004730:Endothelium, Vascular / Q000201:enzymology / Q000276:immunology*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D007249:Inflammation / Q000235:genetics*; D008055:Lipids / Q000737:chemistry; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000201:enzymology / Q000276:immunology*; D011651:Pulmonary Artery / Q000201:enzymology / Q000276:immunology; D011652:Pulmonary Circulation; D051381:Rats; D017207:Rats, Sprague-Dawley; D019076:Transgenes / Q000276:immunology
chemical_list: D008055:Lipids; D004247:DNA; D002795:Choline O-Acetyltransferase; D000469:Alkaline Phosphatase