Retroviral gene transfer into the intestinal epithelium.
The epithelial cells of the gastrointestinal tract may be attractive targets for somatic gene therapy. In these studies, we have used rats and mice to explore the feasibility of gene transfer into the small intestinal epithelium using retroviral vectors. The first series of experiments was conducted in mature Sprague-Dawley rats using an ecotropic retroviral vector that has bacterial beta-galactosidase (beta-Gal) as the reporter gene. The vector was introduced into the lumen of ligated segments of terminal ileum. After a 4-hr exposure period, the ligatures were removed. Sham-operated animals were subjected to the same ligation procedure but received only tissue culture medium in the ligated segment. All animals were sacrificed 6 days later, and tissue from both the experimental segment and an upstream control segment was assessed for cytoplasmic beta-Gal activity using X-Gal histochemistry. Expression of the reporter gene was observed in the crypt epithelium of tissue exposed to the vector. In the villus epithelium, high background staining precluded accurate assessment of reporter gene expression. To obviate the latter problem, we sought an alternative reporter gene for which there would be no background staining in control animals. We repeated the experiments with beta-glucuronidase as the reporter gene in MPS VII mutant mice, which are devoid of this enzyme. In these studies, ileal segments exposed to the vector demonstrated expression of the reporter gene in both the crypt and villus epithelium 4 days after exposure. These results indicate that genes can be transferred into the intestinal epithelium using retroviral vectors introduced luminally.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030-3498, USA.
C
C
Lau
H E
HE
Soriano
F D
FD
Ledley
M J
MJ
Finegold
J H
JH
Wolfe
E H
EH
Birkenmeier
S J
SJ
Henning
D000818:Animals; D004848:Epithelium / Q000187:drug effects / Q000821:virology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000494:pharmacology; D005966:Glucuronidase / Q000235:genetics; D007082:Ileum / Q000601:surgery / Q000648:ultrastructure / Q000821:virology; D007422:Intestines / Q000187:drug effects / Q000648:ultrastructure / Q000821:virology*; D051379:Mice; D008815:Mice, Inbred Strains; D051381:Rats; D012190:Retroviridae / Q000235:genetics*; D013997:Time Factors; D014779:Virus Replication / Q000187:drug effects; D001616:beta-Galactosidase / Q000235:genetics
D001616:beta-Galactosidase; D005966:Glucuronidase