Adenovirus-mediated transfer of human lipase complementary DNA to the gallbladder.
OBJECTIVE
Despite many improvements in current therapy, exocrine pancreatic insufficiency remains a significant problem in cystic fibrosis. To establish a new therapy for exocrine pancreatic insufficiency, the feasibility of transferring the human pancreatic lipase complementary DNA to the gallbladder as a possible target using a recombinant adenovirus vector was evaluated.


METHODS
The adenovirus vector AdCMV. Lip was constructed using the cytomegalovirus immediate early promoter to drive the human pancreatic lipase complementary DNA. In vitro infection of the human gallbladder epithelial cell line HS-181 and ex vivo infection of the sheep gallbladder with AdRSV. beta-gal (an adenovirus vector containing the Escherichia coli lacZ [(beta-galactosidase] gene) or AdCMV. Lip were evaluated.


RESULTS
The supernatant from AdCMV. Lip-infected HS-181 showed the secretion of active lipase for at least 2 weeks in vitro. The epithelium of gallbladder infected with AdRSV.beta-gal ex vivo showed the expression of the beta-galactosidase. The fluid from the gallbladder lumen infected with AdCMV. Lip showed the increased lipase activity.


CONCLUSIONS
These observations show that an adenovirus vector can transfer a human pancreatic enzyme in vitro and ex vivo, suggesting the feasibility of in vivo gene therapy for exocrine pancreatic insufficiency in cystic fibrosis.
Maeda
H
H
Pulmonary Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland.
Danel
C
C
Crystal
R G
RG
D000256:Adenoviridae / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005240:Feasibility Studies; D005704:Gallbladder / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D018014:Gene Transfer Techniques; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000201:enzymology; D008049:Lipase / Q000235:genetics*; D010179:Pancreas / Q000201:enzymology; D012756:Sheep; D001616:beta-Galactosidase / Q000378:metabolism
D018076:DNA, Complementary; D008049:Lipase; D001616:beta-Galactosidase