Aerosol delivery of lipid:DNA complexes to lungs of rhesus monkeys

• Published in: Pharmaceutical research Vol. 15; no. 5; pp. 671 - 679
• Main Authors: MCDONALD, R. J, LIGGITT, H. D, ROCHE, L, NGUYEN, H. T, PEARLMAN, R, RAABE, O. G, BUSSEY, L. B, GORMAN, C. M
• Format: Journal Article
• Language: English
• Published: New York, NY Springer 01.05.1998; Springer Nature B.V
• Subjects: Adenoviruses; Aerosols; Animals; Biological and medical sciences; Cholesterol; Cystic fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator - administration & dosage; Cystic Fibrosis Transmembrane Conductance Regulator - analysis; Cystic Fibrosis Transmembrane Conductance Regulator - genetics; DNA - administration & dosage; Drug Delivery Systems; Drug dosages; Female; Gene therapy; Gene Transfer Techniques; General pharmacology; Humans; Immunohistochemistry; Inflammation; Laboratory animals; Lipids; Lipids - administration & dosage; Lung - chemistry; Lung - cytology; Lung - metabolism; Lungs; Macaca mulatta; Medical sciences; Medical screening; Monkeys & apes; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Plasmids - administration & dosage; Plasmids - genetics; Polymerase Chain Reaction; Primates; Proteins; RNA, Messenger - metabolism; Toxicity; Complexation; Pharmaceutical technology; Lung; Monkey; Lipids; Gene expression; Respiratory system; In vitro; Vertebrata; Mammalia; Gene; Animal; DNA; Primates; Dosage form; Active ingredient; Cystic fibrosis transmembrane conductance regulator; Aerosol batcher; Release
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1023/A:1011902532163

The potential use of aerosol delivery for non-viral gene therapy was tested by nebulization of lipid:DNA complexes to the lungs of rhesus monkeys. Four female rhesus monkeys were dosed with lipid:DNA formulations via aerosol inhalation, where the DNA coded for the human Cystic Fibrosis Transmembrane Conductance Regulator (hCFTR) protein. Delivery of DNA was determined in lung samples by polymerase chain reaction (PCR) by qualitative and quantitative methods. Transgene specific messenger RNA was measured by reverse transcriptase PCR (RT-PCR) and protein expression and localization were evaluated by immunohistochemistry (IHC). Approximately four mg of DNA, complexed with cationic lipid 1.2-dimyristoyl-sn-glycero-3-ethylphosphatidylcholine (EDMPC) and cholesterol were delivered to the lungs of animals by airjet nebulizer. Three days after dosing, tissue samples from the lung were collected and shown to have vector specific DNA, RNA and the presence of CFFR protein. Specifically, the hCFTR protein was distributed widely, although non-uniformly, throughout airway epithelium being located on the apical surface of epithelial cells. Importantly, no adverse clinical effects were observed and the lungs showed no histological abnormalities or signs of acute inflammation. This study shows that lipid:DNA formulations based on EDMPC and cholesterol can be administered to primates by nebulization resulting in measurable expression of the hCFTR protein. The absence of inflammation is also encouraging and such systems may have utility for delivery of genes to the lungs for the treatment of a variety of pulmonary diseases including cystic fibrosis.