vector systems are limited. 
1.9 Progress Report - New York/NIH Gene Therapy Study. The principal in- 
vestigator (RGC) is the principal investigator of an ongoing human gene ther- 
apy study utilizing a replication deficient adenovirus vector (AdCFTR) to 
transfer the normal human CFTR cDNA to the nasal and airway epithelium of 
individuals with CF (Rockefeller University IRB and Biosafety Committee RCR- 
029-0394, NIH DNA Recombinant Advisory Committee 9212-034, Food and Drug Ad- 
ministration BB IND 4855). To date, 7 individuals have been treated (see Ap- 
pendix Al . 1 for details regarding sites of administration, doses, volumes; 
Appendix Al . 2 for details regarding biologic efficacy; Appendix Al . 3 for de- 
tails regarding shedding; Appendix Al.4 for details regarding immunity; and 
Appendix Al . 5 for details regarding safety. In one of these individuals (2A, 
2xl0 9 pfu administered to the airways in a volume of 20 ml) , administration of 
the vector to the airways was associated with a transient local and systemic 
syndrome likely mediated by inflammation induced by the vector. The details of 
this syndrome are in Appendices Al.l-Al.5. 
2. AdgvCFTR.10. The adenovirus vector AdcvCFTR.lO is based on the Ad5 genome 
(Appendix A2.1). It is missing the Ela and most of the Elb sequence, and thus 
is replication deficient, and is missing the majority of the E3 sequence. 
AdcvCFTR.10 contains an expression cassette that includes the cytomegalovirus 
early proraoter/enhancer , an artificial splice sequence, followed by the normal 
human CFTR cDNA and then the SV40 stop/polyA sequences. The infectious vector 
is produced in the 293 human embryonic kidney cell line which supplies the Ad5 
sequences in trans (59,63). Following production, the vector is purified by a 
combination of multiple CsCl' gradient centrifugation steps and dialysis (59). 
The vector is stored frozen in a glycerol - salt solution. 
2.1 Construct. The left end of the AdcvCFTR-lO construct has been sequenced 
through the region of recombination with Ad5 that follows the expression cas- 
sette. The sequence confirms the fidelity of the expression cassette (Appendix 
A2 . 2 ) . 
2.2 Pre-Clinical Studies. In regard to in vitro efficacy, similar to Ad vec- 
tors of similar overall design (59; NIH DNA Recombinant Advisory Committee 
9212-034) in vitro studies with HeLa and the HS24 human lung carcinoma cell 
lines and an epithelial cell line derived from individuals with CF demonstrat- 
ed that AdcvCFTR.10 directed the expression of CFTR mRNA and protein, and cor- 
rected the defect in cAMP-mediated Cl' secretion that characterizes cystic 
fibrosis (Appendix A2.3). In regard to in vivo efficacy, a variety of studies 
with adenovirus vectors similar in design to AdsvCFTR . 10 demonstrated that in 
cotton rats and non-human primates, administration of the vector to the respi- 
ratory tract results in expression of the normal human CFTR CDNA in the airway 
epithelium at the mRNA and protein levels (NIH DNA Recombinant Advisory Com- 
mittee 9212-034; Food and Drug Administration BB IND 4855; 59, 64). Studies 
with respiratory tract administration of Ad^yCFTR-lO to cotton rats demon- 
strated human CFTR cDNA expression in the lung at the mRNA and protein levels 
(Appendix A2.4). In regard to previous in vivo safety, a variety of studies 
with adenovirus vectors similar in construction to AdcvCFTR.10 demonstrated 
that in cotton rats and non-human primates, administration of the vector to 
the respiratory tract results in dose -dependent inflammation in the walls of 
airways, blood vessels and alveoli, with no clinically apparent sequela to the 
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