Protocol 
RAC Application 
Kenneth L. Brigham, M.D. 
Specific Aims 
Wc wish to determine whether a human alpha-1 antitrypsin transgene in an expression 
plasmid vector can be introduced into the cells of the respiratory tract of humans using a 
cationic liposome delivery system and to determine the magnitude and time course of 
expression of the transgene. In addition, we will determine the structural effects of delivery 
of plasmid/cationic liposome complexes on the lungs and on nasal epithelium. Specifically, we 
propose; 
1. To determine whether delivery of a plasmid containing the cDNA for human alpha-1 
antitrypsin complexed to cationic liposomes to the nasal mucosa of patients with alpha-1 
antitrypsin deficiency will result in expression of the alpha-1 antitrypsin transgene in nasal 
epithelial cells and in secretion of the alpha-1 antitrypsin protein into nasal fluid. 
2. To define the time course of expression of a human alpha-1 antitrypsin transgene 
delivered to the nasal mucosa of patients with alpha-1 antitrypsin deficiency using cationic 
liposomes. 
3. To determine effects of a single administration of plasmid/cat ionic liposome complex 
on the histological appearance of human nasal epithelial cells. 
4. To de termine whether deli ve ry of a plasmid containing the cDNA for human alpha-1 
antitrypsin complexed to cationic liposomes to the lower respiratory tract of patients scheduled 
for elective pneumonectomy will result in expression of the alpha-1 antitrypsin transgene in 
airway epithelium and/or other lung cells and synthesis and secretion of the alpha-1 
antitrypsin protein by the lungs. 
5. To determine effects of a single administration of plasm id /cat ionic liposome complex 
to the lower respiratory tract on the histological appearance of the lungs. 
Background and Significance 
Gene therapy in the lungs Most research in gene therapy in the lungs has focused on 
the two heritable disorders, alpha-1 antitrypsin deficiency and cystic fibrosis. Since the 
discovery of the cystic fibrosis transmcmbranc conductance regulator gene (CFTR), the 
principal driving force toward clinical application of gene therapy to the lungs has been cystic 
fibrosis. 
The most extensively studied technology for in vivo delivery of transgenes to the lungs 
involves the use of replication deficient adenovirus vectors. Such vectors can deliver reporter 
genes as well as genes encoding cither AAT or CFTR (1,2) to the lungs of animals. Based on 
that work, there are now several phase I clinical trials attempting to deliver the CFTR to 
patients with cystic fibrosis either intranasally or via bronchoscopy to the lower respiratory 
tract. Early reports from studies in humans indicate that nasal instillation is safe (3). 
There arc several potential problems with the adenovirus vector. Some investigators 
have seen an acute inflammatory response to the vector when delivered into the airways in 
animals (4). At least one patient in the clinical studies with intra-airway administration 
appears to have had a clinically important inflammatory response (5). Although replication 
deficient, the vector does involve delivery of viral proteins resulting in an immune response 
which could limit long term usefulness. 
Recombinant DNA Research, Volume 19 
[175] 
