"A Phase I Study of Gene Therapy of Cystic Fibrosis Utilizing a Replication Deficient 
Recombinant Adenovirus Vector to Deliver the Cystic Fibrosis Transmembrane Conductance 
Regulator Gene to the Airways. " 
The purpose of this study is to evaluate the safety and efficacy of gene therapy of cystic 
fibrosis with a replication deficient AE1,AE3 deleted adenovirus that directs expression of the 
normal CFTR mRNA and CFTR protein in mammalian cells. The virus, AvlCF2 will be 
administered to the upper (nasal) and lower (lobar bronchial) respiratory tract of patients with 
cystic fibrosis. This protocol is designed to demonstrate 1) the expression of normal CFTR 
mRNA in vivo, 2) the synthesis of CFTR protein and 3) the correction of epithelial cell cAMP 
dependent Cl' permeability associated with cystic fibrosis. Potential toxic effects of AvlCF2 will 
be monitored by measuring clinical, radiographic, physiologic and biochemical parameters. The 
"pharmacokinetics" or longevity of expression and ability to re-infect the respiratory tract with 
AvlCF2 will be determined. Systemic and local immunologic consequences of AvlCF2 
infection, the time of viral survival, and the potential for recombination or complementation of 
the virus to produce a replicating virus will be monitored in assessing the safety of the AvlCF2 
recombinant virus. The safety and potential risks of exposure of personnel and cohorts of the 
patients after AvlCF2 administration will be assessed. 
Consenting male or female patients of age 18 or greater with FVC >40% and documented 
homozygote A508 genotype will be recruited from the Cystic Fibrosis Center at Children’s 
Hospital Medical Center, Cincinnati. Patients will be admitted to the Research Center, 
Children’s Hospital after undergoing a two week period of intensive pulmonary toilet including 
antibiotics, postural drainage, aerosols, mucolytics and/or DNAse therapy (pending FDA 
approval of DNAse). Pulmonary function tests will be evaluated pre and post "cleanout." 
AvlCF2 will then be administered to specific regions of the inferior nasal turbinate, each patient 
group receiving 5 x 10 7 , 5 x 10 8 or 5 x 10 9 PFU to defined areas of the nasal epithelium. CFTR 
mRNA, CFTR protein and Cl' permeability will be assessed in nasal epithelial cells. Three days 
later, the patient will receive the AvlCF2 in the bronchus of the right or left lower lobe on 
bronchoscopic examination. Patient groups will receive 10 10 , 10 11 or 10 12 PFU per dose in the 
lung. Clinical, radiographic, physiologic and cellular evidence of efficacy and toxicity will be 
assessed post administration. Viral clearance and replication will be assessed and the patient 
discharged when clear of infectious AV,CF virus. 7.5 weeks after initial treatment, the patient 
will be readmitted and the presence of CFTR mRNA and lung function re-assessed. The same 
dose of virus previously administered to a particular patient will then be readministered to 
evaluate the potential for reinfection and expression of the CFTR mRNA to the lung epithelium. 
The virus, AvlCF2, was constructed by Bruce Trapnell at Genetic Therapy, Inc., 
Gaithersberg, Maryland, from genetic material obtained from the adenovirus serotype 5. In 
AvlCF2, human CFTR mRNA is driven by the RSV, (Rous sarcoma promoter-enhancer) and 
terminates with SV40 poly A signals. The virus harbors deletions in the El (A El) and E3 
regions (A E3) deleted to block replication and to provide insert space for the incoming cDNA 
respectively. Similar adenoviral constructs have been used to transfer cDNA’s for a variety of 
genes to various mammalian cells in vitro and to lung epithelial tissues in vivo by this laboratory 
and by others. It is hoped that these studies will demonstrate the biologic expression in vivo and 
safety of the AvlCF2 vector as a phase I study and support the long term goal of CFTR gene 
transfer that will lead to physiologically, and ultimately clinically, relevant reconstitution of the 
abnormalities that cause pulmonary disease in cystic fibrosis. 
Recombinant DNA Research, Volume 17 
[353] 
