Human APC gene expression in rodent colonic epithelium in vivo using liposomal gene delivery.
Mutations or loss of the APC tumor-suppressor gene is important for the development of colorectal polyps and cancers, but little is known about the function of this gene in normal tissue. To study the role of APC and other genes in colonocytes in vivo, a system was developed whereby transient expression of genes is established in normal rodent colonic epithelium, using liposomal gene delivery by rectal catheter infusion. Expression of a beta-galactosidase reporter gene and of the human APC gene under a constitutive promoter is demonstrated. A high efficiency of transfection is maintained, with close to 100% of epithelial cells expressing the introduced gene. Expression is transient and does not persist beyond 4 days, consistent with the normal turnover time of gut epithelium, but it can be maintained by repeated treatments. Human APC was expressed for three weeks under these conditions at approximately one-tenth the level of the endogenous APC gene, and no toxicity was observed beyond that attributed to repeated rectal enemas. These results reveal that in vivo expression of exogenous gene is feasible using a liposomal delivery system and suggest a method to further study the physiologic role of APC or other genes in the interrelated process of colonic epithelial proliferation and differentiation.
Westbrook
C A
CA
Department of Medicine, University of Chicago, IL 60637.
Chmura
S J
SJ
Arenas
R B
RB
Kim
S Y
SY
Otto
G
G
D000818:Animals; D001483:Base Sequence; D003106:Colon / Q000737:chemistry* / Q000201:enzymology; D015870:Gene Expression / Q000235:genetics*; D018014:Gene Transfer Techniques; D017491:Genes, APC; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry / Q000201:enzymology; D008081:Liposomes / Q000737:chemistry*; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley; D001616:beta-Galactosidase / Q000096:biosynthesis
D008081:Liposomes; D001616:beta-Galactosidase