Gene Therapy for CF using Cationic Liposome Mediated Gene Transfer: Phase I Trial 
a. Toxicity . 
1) Serial assessment by history and physical examination, and measurements 
including blood chemistry, complete blood count, pulmonary function tests, 
oximetry, EKG, chest x-ray and sputum culture will be performed in order to 
evaluate individuals in this study for evidence of local (nasal), pulmonary or 
systemic toxicity. Serum will also be studied for evidence of humoral immune 
responses to administered lipid or DNA. (1) 
2) Direct visualization of nasal mucosa for evaluation of inflammation or other 
changes will be accomplished using nasal endoscopy. This procedure will be 
performed in collaboration with Dr. Brian Wiatrak (Department of Pediatric 
Otolaryngology). After local anesthesia with 2% Xylocaine a flexible nasal 
endoscope will be used to visually observe the inferior turbinate area for evidence 
of edema, exudate, or purulence following lipid or lipid/DNA administration. 
3) Inflammation at the site of CFTR gene administration and mock (no CFTR) 
administration (in the contralateral nostril) will be studied using nasal biopsy, 
nasal brushings and washings, as described below: 
aa. Nasal biopsy . Biopsies of nasal epithelium will be performed in order to 
evaluate inflammation and other cytopathic effects due to lipid/DNA 
conjugate administration. This procedure will be performed by Dr. B. 
Wiatrak as follows: The nasal cavity will initially be decongested with 2% 
Pontocaine/.25% Phenylephrine on cotton pledgets. The site of the biopsy 
will be the inferior aspect of the inferior turbinate with an area of 
approximately 0.5cm 2 . The area will be injected submucosally with 1% 
Lidocaine with 1:100,000 Epinephrine. The mucosal biopsy will then be 
excised using a #11 scalpel blade and hemostasis obtained by placing 25 % 
Neo-Synephrine-soaked pledgets. Biopsies will be assessed for evidence 
of mucosal disruption, cytolysis, or inflammation using light microscopy. 
Electromicroscopy will also be performed for evidence of cytopathic 
effects. Tylenol will be given as required for analgesia. 
Although lipid and DNA are generally poorly immunogenic, in order to determine whether a systemic 
humoral antibody response develops in animals given lipid/DNA conjugates, a series of experiments will be performed 
in which rats given 500/rg of plasmid DNA conjugated to 400pg DMRIE/DOPE in a final volume of 800^1 (as in Figure 
4, Preclinical Data Section). Rats will be bled at 1,2,3, and 4 weeks following intratracheal administration. Serum from 
these bleeds (as well as from a pre-DNA administration bleed) will be used in an ELISA assay in which 1/rg samples 
of lipid/DNA conjugates per microtiter well are overlaid with dilutions of serum between 1:10 and 1:10’. Goat anti-rat 
IgG conjugated to alkaline phosphatase will be used to detect any antibody arising from lipid/DNA conjugates using a 
standard ELISA protocol (for example, see Sorscher, et al . Am J Physiol (Cell Physiol) 255:c835-c843, 1988). Similar 
experiments (using an antihuman IgG conjugated to alkaline phosphatase) will be performed as part of the human study 
described here. 
Recombinant DNA Research, Volume 18 
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