Aerosol and lobar administration of a recombinant adenovirus to individuals with cystic fibrosis. II. Transfection efficiency in airway epithelium.
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.
Perricone
M A
MA
Genzyme Corporation, Framingham, MA 01701, USA. michael. perricone@genyme.com
Morris
J E
JE
Pavelka
K
K
Plog
M S
MS
O'Sullivan
B P
BP
Joseph
P M
PM
Dorkin
H
H
Lapey
A
A
Balfour
R
R
Meeker
D P
DP
Smith
A E
AE
Wadsworth
S C
SC
St  George
J A
JA
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
C505032:CFTR protein, human; D004274:DNA, Recombinant; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator