Genzyme Corporation Ad2/CFTR-2 Gene Transfer Protocol: Single 
One Kendall Square, Cambridge, MA 02139 Aerosol Administration to the Lung of CF Patients 
ADENOVIRUS-MEDIATED GENE TRANSFER FOR CYSTIC FIBROSIS: SAFETY OF SINGLE AEROSOLIZED 
ADMINISTRATION TO THE LUNG 
Investigators: Henry Dorkin, M.D. of the New England Medical Center and Allen Lapey, M.D. of 
Massachusetts General Hospital 
Co-investigators: Joseph Oren, M.D. of the New England Medical Center and Patricia Joseph, M.D. of 
Massachusetts General Hospital 
Protocol 
INTRODUCTION 
Cystic fibrosis (CF), is a lethal autosomal recessive disease resulting from mutations in the gene encoding the cystic 
fibrosis transmembrane conductance regulator (CFTR) protein, a protein which forms a chloride channel that is 
regulated by phosphorylation and by intra-cellular nucleotides [1-3]. Mutations in the CFTR gene cause a loss of CFTR 
chloride channel activity and thus contribute to the hallmark of the disease: defective electrolyte transport by affected 
epithelia [4-6] . Our current understanding of the structure and function of CFTR suggests that gene transfer could 
represent an important advance in treatment. 
The feasibility of gene transfer to lung cells was initially demonstrated by the finding that expression of the cDNA for 
wild-type CFTR corrected the chloride channel defect in cultured CF airway epithelia cells [7] . Subsequently, 
recombinant adenovirus vectors have been constructed and tested in cell culture models and in animals for their ability 
to deliver CFTR cDNA to airway epithelial cells. The results of these studies are encouraging in terms of the ability of 
adenovirus vectors to transfer and to express CFTR. The safety record of their administration at doses predicted to be 
relevant for the treatment of patients with CF [8], is important and thus far satisfactory. 
Based on the results of these preclinical studies, a clinical strategy for CF gene therapy was planned. Because patient 
safety is the primary concern while any questions about the safety of adenovirus vectors remain, we have adopted a 
conservative clinical trials approach, one in which safety and efficacy of adenovirus vectors are first assessed in the 
upper airways before attempting lower airway (i.e. lung) administration. In our first clinical trial, ’Cystic Fibrosis Gene 
Therapy: In Vivo Safety and Efficacy in Nasal Epithelium (ORDA #9212-036), a single dose of an adenovirus-based 
vector encoding CFTR (Ad2/CFTR-1) was administered to the nasal respiratory epithelium of 4 patients. Although the 
first 2 patients experienced some local inflammation attributable to the device used for nasal instillation, there were no 
adverse events observed in these 4 patients associated with the use of Ad2/CFTR-1 [8]. Evidence of transient 
restoration of CFTR channel activity was obtained. 
Our second clinical trial, ’Adenovirus-Mediated Gene Transfer for Cystic Fibrosis: Part A-Safety of Repeat 
Administration in the Nasal Epithelium and Part B-Safety of Repeat Administration in the Maxillary Sinus" (ORDA 
#9312-067) uses the vector Ad2/CFTR-2 to address two critical issues; the effect of repeat vector administration on 
safety and biochemical efficacy, and the potential for achieving clinical efficacy upon restoration of CFTR channel 
function in the upper airways. Repeat doses of the vector are administered to the nasal and sinus respiratory epithelium 
to gain the maximum information while minimizing risk to patients. To date, 2 patients have received 4 administrations 
of Ad2/CFTR-2 (2 x 10 7 , 2 x 10 s , 2 x 10 9 , and 6.6 x 10 9 IU) and two additional patients have received the 2 x 10 7 dose 
to the nasal epithelium with no study drug related adverse effects. Evidence of transient restoration of CFTR channel 
activity has also been obtained. 
In this proposed study, we plan to assess the safety of single dose administration of Ad2/CFTR-2 to the lower airway by 
aerosolization to the lung after testing of the dose in a concurrent protocol for administration to a single lobe of the lung. 
The efficient delivery of gene transfer to patients with CF will be accomplished by aerosol to the airway. In a future study 
we will assess the safety and efficacy of repeat aerosol administration of Ad2/CFTR-2 to the lung. 
STUDY OBJECTIVE 
The objective of this study is to evaluate the safety of a single aerosolized administration of Ad2/CFTR-2 to the lung of 
cystic fibrosis patients. 
Recombinant DNA Research, Volume 20 
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