title: Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. II. Transfection efficiency in airway epithelium.
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.
authors.affiliation: Genzyme Corporation, Framingham, MA 01701, USA. michael. perricone@genyme.com
authors.forename: M A
authors.initials: MA
authors.lastname: Perricone
authors.forename: J E
authors.initials: JE
authors.lastname: Morris
authors.forename: K
authors.initials: K
authors.lastname: Pavelka
authors.forename: M S
authors.initials: MS
authors.lastname: Plog
authors.forename: B P
authors.initials: BP
authors.lastname: O'Sullivan
authors.forename: P M
authors.initials: PM
authors.lastname: Joseph
authors.forename: H
authors.initials: H
authors.lastname: Dorkin
authors.forename: A
authors.initials: A
authors.lastname: Lapey
authors.forename: R
authors.initials: R
authors.lastname: Balfour
authors.forename: D P
authors.initials: DP
authors.lastname: Meeker
authors.forename: A E
authors.initials: AE
authors.lastname: Smith
authors.forename: S C
authors.initials: SC
authors.lastname: Wadsworth
authors.forename: J A
authors.initials: JA
authors.lastname: St  George
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000280:Administration, Inhalation; D000293:Adolescent; D000328:Adult; D001999:Bronchoscopy; D003550:Cystic Fibrosis / Q000628:therapy*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004274:DNA, Recombinant; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007322:Instillation, Drug; D008297:Male; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification; D020545:Respiratory Mucosa / Q000378:metabolism*; D013997:Time Factors; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: C505032:CFTR protein, human; D004274:DNA, Recombinant; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator