Scientific Abstract 
RAC Application 
Kenneth L. Brigham, M.D. 
Scientific Abstract 
SCIENTIFIC ABSTRACT 
We propose initial studies in humans with the long term goals of using gene transfer 
techniques for the treatment of patients with acute lung injury (adult respiratory distress 
syndrome— ARDS) and patients with alpha-1 antitrypsin (AAT) deficiency. We propose to use 
cationic liposomes to deliver a plasmid DNA construct with the cDNA for human alpha-1 
antitrypsin driven by a cytomegalovirus promoter. Two different patient protocols are proposed 
using the exact same DNA and liposome preparations. In patients scheduled for elective 
pulmonary resection, we will instill plasmid/liposome complexes into a subsegment of the lung 
to be removed through a fiberoptic bronchoscope and obtain samples of the transfected portion 
and of the non-transfected portion of the lungs at the time of surgery. By 
immunohistochemistry, in situ hybridization, western and northern analysis, we will assess the 
amount and location of transgene expression in the lungs. Histological studies will also 
determine the effects of the intervention on lung structure. In patients withalpha-1 antitrypsin 
deficiency, we will instill plasmid/liposome complexes into one nostril and take serial nasal 
lavages from each nostril, measuring alpha-1 antitrypsin by ELISA in the lavage samples to 
assess expression of the AAT transgene. Cells obtained by nasal scraping will also be examined 
by immunohistochemical staining and [n situ hybridization to assess transgene expression. 
Histological appearance of the cells will provide information about effects of the intervention 
on nasal epithelial cell structure. Because proteolytic events, principally a consequence of the 
inflammatory process, appear to be important in the pathogenesis of acute lung injury and of 
chronic pulmonary disease in patients with AAT deficiency (emphysema), development of safe 
and efficacious systems for delivering the AAT gene to respiratory tract cells might provide a 
new form of therapy for patients with these diseases. 
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Recombinant DNA Research, Volume 19 
