Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. II. Transfection efficiency in airway epithelium
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. ... show 3 more
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Student Authors
Faculty Advisor
Academic Program
UMass Chan Affiliations
Document Type
Publication Date
Keywords
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
Subject Area
Embargo Expiration Date
Link to Full Text
Abstract
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.
Source
Hum Gene Ther. 2001 Jul 20;12(11):1383-94. Link to article on publisher's site