In utero AAV-mediated gene transfer to rabbit pulmonary epithelium.
In utero intra-amniotic administration of adeno-associated virus (AAV) for treatment of cystic fibrosis (CF) has the potential to be an efficient way to target the rapidly dividing undifferentiated cells of the fetal pulmonary epithelium, while simultaneously treating other tissues involved in CF (such as the intestines), but has never before been studied. Intra-amniotic administration of 1x10(12) particles of AAV-luciferase vector to 110 fetal rabbits at 24-25 days gestation resulted in transgene expression in amniotic membranes, trachea, and pulmonary epithelium. The highest level of transgene expression was found in amniotic membranes. Transgene expression peaked in the lungs 10 days after vector delivery, decreased at day 17, and was no longer detectable after 24 days. The number of pulmonary cells transduced was approximately 1 in 500 and immunohistochemical analysis showed expression in varying cell types, including alveolar cells. Transgene expression was not detected in fetal rabbit intestines, skin or liver, nor in maternal ovaries or liver. Intra-amniotic administration of AAV does not result in the tissue inflammation and fetal loss previously documented with in utero adenoviral administration, and results in high levels of transgene expression in amniotic membranes with lower levels in fetal pulmonary epithelium.
Boyle
M P
MP
Department of Medicine, The Johns Hopkins University School Of Medicine, Baltimore, Maryland 21205, USA. mboyle@mail.jhmi.edu
Enke
R A
RA
Adams
R J
RJ
Guggino
W B
WB
Zeitlin
P L
PL
D000650:Amnion / Q000378:metabolism; D000818:Animals; D000229:Dependovirus / Q000235:genetics* / Q000502:physiology; D005260:Female; D005333:Fetus / Q000502:physiology; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry; D007267:Injections; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011817:Rabbits; D020545:Respiratory Mucosa / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D012394:Rosaniline Dyes / Q000378:metabolism; D014132:Trachea / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D019076:Transgenes
D012394:Rosaniline Dyes; C035906:Fast Green; D008156:Luciferases