Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. II. Transfection efficiency in airway epithelium
Department of Pediatrics
Adenoviridae; Administration, Inhalation; Adolescent; Adult; Bronchoscopy; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; DNA, Recombinant; Female; Genetic Vectors; Humans; In Situ Hybridization, Fluorescence; Instillation, Drug; Male; Polymerase Chain Reaction; RNA, Messenger; Recombinant Proteins; Respiratory Mucosa; Time Factors; Transduction, Genetic; *Transfection
Allergy and Immunology | Pediatrics | Respiratory Tract Diseases
A phase I clinical trial was conducted in which recombinant adenovirus containing the cystic fibrosis trans-membrane regulator (CFTR) (Ad2/CFTR) was administered by bronchoscopic instillation or aerosolization to the lungs of cystic fibrosis (CF) patients. In this paper, we evaluate the efficiency of Ad2/CFTR-mediated transduction of bronchial airway cells. The ability of an Ad2/CFTR vector to transduce airway cells was first evaluated in patients to whom the vector was administered by bronchoscopic instillation. Cells at the administration site were collected 2 days after treatment by bronchoscopic brushing. Ad2-specific CFTR DNA was detected in four of five individuals by PCR, and Ad2-specific CFTR RNA was detected in three of five individuals by RT-PCR. Ad2/CFTR-mediated transduction of airway epithelial cells was then determined in CF individuals receiving this vector by aerosol inhalation. Ad2-specific CFTR DNA was detected in 13 of 13 individuals 2 days after aerosolization, and in 3 of 5 individuals 7 days after aerosolization. Ad2-specific RNA was detected in 4 of 13 individuals on day 2, but was not detected in the 5 individuals tested on day 7. The percentage of airway epithelial cells containing nuclear-localized vector DNA was < or =2.4% as determined by fluorescence in situ hybridization (FISH). However, in some cases, a high percentage of nonepithelial mononuclear cells or squamous metaplastic epithelial cells was infected with the adenoviral vector. In conclusion, aerosol administration is a feasible means to distribute adenoviral vectors throughout the conducting airways, but improvements in adenovirus-mediated transduction of airway epithelial cells are necessary before gene therapy for CF will be effective.
DOI of Published Version
Hum Gene Ther. 2001 Jul 20;12(11):1383-94. Link to article on publisher's site
Human gene therapy
Perricone, Michael A.; Morris, James E.; Pavelka, Karen; Plog, Malinda S.; O'Sullivan, Brian P.; Joseph, Patricia M.; Dorkin, Henry L.; Lapey, Allen; Balfour, Rosemary; Meeker, David P.; Smith, Alan E.; Wadsworth, Samuel C.; and St. George, Judith A., "Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. II. Transfection efficiency in airway epithelium" (2001). Pulmonary and Allergy. 4.