Adenovirus-Mediated Gene Transfer for Cystic Fibrosis: Safety of Single Administration In the Lung 
COVER SHEET FOR ACCELERATED REVIEW OF A HUMAN GENE TRANSFER EXPERIMENT 
A. BACKGROUND 
Our proposed study, “Adenovirus Mediated Gene Transfer for Cystic Fibrosis: Safety of Single Administration 
in the Lung” will be performed by Henry Dorkin, M.D. of the New England Medical Center (Tufts University) and 
Allen Lapey, M.D. of the Massachusetts General Hospital (Harvard Medical School). 
a VECTOR, TARGET CELL, AND TRANSDUCTION PROCEDURES 
1. The replication defective type 2 adenovirus vector, Ad2/CFTR-2 will be used to deliver the human cystic 
fibrosis transmembrane conductance regulator (CFTR) gene. 
Z The Ad2/CFTR-2 vector was reviewed by the RAC in considering protocol #9312-067 entitled, "Adenovirus 
Mediated Gene Transfer for Cystic Fibrosis: Part A. Safety of Dose and Repeat Administration in the Nasal 
Epithelium and Part B. Clinical Efficacy in the Maxillary Sinus”. This protocol is being performed by Dr. 
Michael Welsh at the University of Iowa and by Dr. Bonnie Ramsey at the University of Washington. 
3. Ad2/CFTR-2 will be manufactured, tested and supplied by Genzyme Corporation. 
4. The target cells to be treated are epithelial cells lining the surface of the lower airway. 
5. The rate of transduction has not been established. Because adenovirus DNA is predominantly not integrated 
and because the percentage of target cells that contain DNA and the copy number in those cells is dependent 
on the MOI, we have not examined in detail the number of target cells that contain added DNA. Instead, we 
have measured the expression of the DNA. 
6. It is expected that the level of gene expression will be dependent on the multiplicity of infection or dose of 
vector. Results of in vitro and in vivo studies performed to assess the efficacy of Ad2/CFTR-2 were 
reported in our December, 1993 RAC application. 
7. Gene expression has been assessed by in vitro cell culture models, by in vitro SPQ and Ussing chamber 
assays that assess biological activity of the expressed gene and using in vivo animal models. 
& We utilize a HeLa cell/A549 cell assay to detect replication competent adenovirus (RCA). 
9. When assayed by the most sensitive methods available, replication competent adenovirus has been detected 
in some lots of Ad2/CFTR-2. In clinical trials we are using only lots of product that pass the RCA test 
referred to above. Pending FDA approval, we may use lots of Ad2/CFTR-2 that contain low levels of RCA. 
10. The HeLa cell/A549 cell assay used to detect RCA has a limit of detecting one infectious unit of RCA in the 
presence of 2.5x1 0 9 IU of Ad2/CFTR-2. 
C. CLINICAL PROTOCOL 
■ 
1. As noted in B.7. above, preclinical efficacy data has been generated using both in vitro and in vivo models 
Z The primary objective of the study is to assess the safety of delivering Ad2/CFTR-2 to lower airways in the 
lung by lobar instillation (through a bronchoscope) and by aerosolization to the whole lung. A secondary 
objective of the study is to assess gene expression following treatment. 
3. This protocol is not identical to another RAC approved protocol. 
4. This protocol is similar to the on-going clinical evaluation of Ad2/CFTR-2 as it is primarily a safety study 
during which biochemical efficacy will be assessed following administration of single, escalating doses of 
vector. The differences are that the target organ is the lung and the mode of delivery is to be by lobar 
instillation and aerosolization to the whole lung. j 
D. LOCAL COMMITTEE APPROVALS 
1. The study has been unconditionally approved by the IBC of the New England Medical Center. The IBC of 
Harvard Medical School made requests for increased monitoring, for an opportunity to review the final 
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