Prenatal orogastric gene delivery results in transduction of the small bowel in the fetal rabbit.
BACKGROUND
Gene therapy for meconium ileus, or other genetic diseases involving the gastrointestinal epithelium, may be possible with prenatal delivery of the CFTR gene to the gastrointestinal tract. Although minimally invasive techniques will probably be used for any future therapy of gastrointestinal disease, it is important to first test this strategy with a reliable animal model.


METHODS
The technique of orogastric fetal gene delivery was assessed using 7 pregnant rabbits (22 days' gestation n = 1, 25 days' gestation n = 4, 28 days' gestation n = 2). Four fetuses from each litter were given an adenoviral vector carrying a marker gene by instilling it into the posterior pharynx with an animal feeding needle (1 x 10(10) particles of ADV/RSV/LacZ suspended in 0.3 ml of saline), with the untreated fetuses serving as control animals.


RESULTS
There were no recoverable fetuses from the does that had surgery at 22 and 28 days (n = 3) due to maternal death (n = 2) and premature delivery (n = 1). Among the 4 does that underwent hysterotomy at 25 days of gestation, 1 underwent cesarean section 2 days after fetal gene delivery and 3 delivered at term, 5 (n = 1) or 6 (n = 2) days following gene delivery. Eleven of the 16 experimental pups and 7 untreated control animals were collected alive, and were sacrificed at delivery for study. Nine of the 11 experimental pups (82%) showed positive blue (LacZ+) nuclei in the small intestine by X-gal staining. No positive cells were found in 7/7 control animals. The presence of the reporter gene LacZ was confirmed in 8/11 (73%) virus-treated pups by PCR with 5/5 control animals negative for LacZ by PCR.


CONCLUSIONS
There was significant maternal and fetal loss related to anesthetic and husbandry issues when surgery was performed at 22 or 28 days of gestation. Based on these preliminary results, we conclude that orogastric gene delivery in the rabbit fetus at 25 days' gestation is an encouraging animal model to study fetal delivery to the gastrointestinal tract.
Wu
Y
Y
Department of Surgery, Baylor College of Medicine, Houston, TX, USA.
Liu
J
J
Woo
S
S
Finegold
M J
MJ
Brandt
M L
ML
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D004274:DNA, Recombinant / Q000032:analysis; D005260:Female; D005333:Fetus / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D015316:Genetic Therapy; D005822:Genetic Vectors; D005865:Gestational Age; D007421:Intestine, Small / Q000196:embryology* / Q000378:metabolism*; D010614:Pharynx / Q000196:embryology; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011817:Rabbits; D001616:beta-Galactosidase / Q000235:genetics*
D004274:DNA, Recombinant; D005819:Genetic Markers; D001616:beta-Galactosidase