Gene Therapy for CF using Canonic Liposome Mediaied Gene Transfer: Phase 1 Trial 
define a 3-5 mm region along the inferior surface of the right inferior turbinate suitable 
for lipid/DNA conjugate administration. This area will be gently rinsed with normal 
saline prior to gene administration. Under direct visualization, a 1 ml volume of 
lipid/DNA formulation (see below) will be administered using a syringe pump, over a 
30 minute period. The patient will then be turned to a right lateral decubitus position and 
a nasal endoscopy used to identify a corresponding contralateral region beneath the left 
inferior turbinate. A 1 ml volume of lipid/DNA solution without CFTR will then be 
administered over 30 minutes. 
4. Lipid/DNA formulations . The lipid formulation which will be used in this study is a 1:1 
molar mixture of DMRIE with DOPE. The DMRIE/DOPE and DNA used in this clinical 
protocol will be generated at Genzyme, Inc., (Framingham, Mass.) under GMP 
guidelines and following FDA-approval and supervision. Lipid and plasmid DNA will 
be packaged desiccated in separate vials and will arrive at the University of Alabama at 
Birmingham by overnight carrier. The vials will each be hydrated with sterile, distilled 
water in a laminar flow hood located in the IV Room of the Pharmacy, Ground Floor of 
The Children's Hospital; this is the usual location of mixing of intravenous solutions for 
patient administration. In vitro evidence suggests that gene transfer using lipid DNA 
conjugates requires optimization of the lipid:DNA base ratio. We will study three patient 
groups (see Table 1, Appendix); each group with a different ratio of CFTR plasmid 
DNA:lipid formulation. CFTR plasmid DNA used in these experiments will be the 
pKCTR, which contains CFTR cDNA under regulatory control of the SV-40 early 
promoter (Figure 7, Preliminary Data Section). Mock (no CFTR) transfections will be 
performed using a vector derived from pKCTR in which CFTR, as well as SV-40 
promoter and polyadenylation sequences have been deleted. 
Based upon experiments using similar proportions of lipid: DNA in vitro , and because the 
range used in this study includes both lipid excess and DNA base excess, this range 
offers the best likelihood of encompassing the appropriate proportion of lipid: DNA for 
optimal gene transfer. Assuming that 1 cm 2 of nasal mucosa has approximately 2x10 s 
cells along the surface epithelium and that the nasal mucosa to be studied will contain on 
the order of 5 times this area (an estimated 10' cells) even at the lowest dose of DNA 
(100 meg) approximately 3x10 s copies of CFTR plasmid DNA will be administered per 
target cell. 
5. Evaluation following lipid- DNA conjugate administration . 
Patients will be monitored in the clinical research center with vital signs every 30 
minutes for the first two hours following gene administration, then on an hourly basis for 
the next six hours, every two hours for the next 16 hours and finally every 6 hours for 
the duration of the admission to the CRC. Admission to the CRC will be for a total of 
five days. During hospitalization all standard cystic fibrosis therapies will continue. In 
order to maximize safety, each patient’s hospitalization will be completed prior to 
proceeding to the next patient. A description of the procedures to be used is given below, 
followed by a flow sheet schedule (see Table 2, Appendix) of the specific tests to be used 
in this protocol. 
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Recombinant DNA Research, Volume 18 
