1 
animals (NIH DNA Recombinant Advisory Committee 9212-034; Food and Drug Ad- 
ministration BB IND 4855; 65). Studies with repeat respiratory tract ad- 
ministration of Ad^vCFTR.10 to cotton rats demonstrated dose -dependent inflam- 
mation after the initial administration, but only minimal inflammation after 
repeat administration, with no clinically- related sequela (Appendix A2.5). 
2.3 Clinical Preparation. The clinical lots of Ad^CFTR.10 will have titers of 
2xl0 9 - 2xl0 10 pfu/ml. Storage will be in glycerol - salt solution at -70°. All 
lots will fulfill all of the in vitro and in vivo safety criteria established 
by the FDA for adenovirus vector preparation, including the criteria of <1 
replication competent adenovirus for the total clinical dose to be delivered. 
3. Overall Design of the Clinical Study. The purpose of this protocol is to 
evaluate the safety and biologic efficacy of repeat administration of the 
AdcvCFTR.10 vector to the epithelium of the large airways of individuals with 
cystic fibrosis. The study is divided into 3 parts. Part A is an ascending 
single dose toxicity/gene expression study to define the pharmacodynamics of 
expression of the normal CFTR cDNA in the airway epithelium following single 
dose administration of the vector to the airways. Part B is designed to eval- 
uate toxicity/gene expression of repeat dosing at the doses showing expression 
in Part A, while part C is designed to evaluate toxicity/gene expression of 
repeat dosing at a higher dose than that in part A. Compared to the Principal 
Investigator's New York/NIH study (Rockefeller University IRB and Biosafety 
Committee RCR-029 -0394 , NIH DNA Recombinant Advisory Committee 9212-034, Food 
and Drug Administration BB IND 4855), the key differences are; (1) a more 
active constitutive viral promoter (cytomegalovirus early promoter/enhancer 
compared to the Ad2 major late promoter/tripartite leaders); (2) the promoter- 
CFTR cDNA expression is 5' -3' toward the left inverted terminal repeat (to 
minimize expression of viral genes distal to the expression cassette stop 
signal); (3) an artificial splice sequence built into the junction of the 
promoter and cDNA (to enhance expression); (4) a limited volume to administer 
the vector via the bronchoscope (<1 ml compared to 20 ml in the original New 
York/NIH study, to minimize alveolar deposition of the vector, and thus mini- 
mize toxicity); (5) additional steps to purify the vector (to minimize vector- 
induced toxicity); (6) administration to multiple airway sites, compared to 1 
site (to maximize the assessment of gene transfer); and (7) repeat administra- 
tion (compared to single administration). At the completion of this protocol, 
a critical question will be answered - can an adenovirus vector safely and ef- 
fectively be used to chronically maintain expression of the normal CFTR cDNA 
in airway epithelial cells of individuals with CF? 
Conceptual details of the design of each part of the protocol can be found in 
Appendix A4 . In parts A and B, prior to administration of the vector, para- 
meters for safety and expression will be evaluated in the control periods 
(baseline and following administration of the vehicle used to suspend the 
vector). In part C, the baseline and vehicle control for part A will be used, 
since part C represents an extension of the dosing in part A. Safety and bio- 
logic efficacy parameters will be evaluated periodically as detailed in sec- 
tion 6-8. The vehicle or vector will be administered at each site via a cathe- 
ter via a fiberoptic bronchoscope in a volume of 100 /xl . 
4. Inclusion/Exclusion Criteria 
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