Somatostatin gene transfer and expression in endothelial cells.
OBJECTIVE
The antiproliferative and antisecretory effects of somatostatin have many potential uses in the clinical setting. Retroviral gene transfer of somatostatin to endothelium is a potential means of local delivery of this peptide to specific vascular beds. This investigation was designed to determine whether transduced endothelial cells (ECs) could produce and post-translationally process somatostatin.


METHODS
Cultured canine venous, rat aortic, and rat microvascular ECs were transfected with retroviruses containing a human somatostatin cDNA or a control beta-galactosidase gene. Total and isoform somatostatin production and uniformity of beta-galactosidase expression were analyzed, as were the effects of somatostatin production on EC proliferation.


RESULTS
Somatostatin-transduced canine venous ECs, but not rat ECs, produced approximately 10 times as much total somatostatin as did control-transfected ECs (450 +/- 32 vs 49 +/- 10 pmol/L, p < 0.05). The predominant isoform of somatostatin produced was somatostatin-14. Production of somatostatin was stable with passage and did not impair the growth of canine ECs. The failure of rat ECs to produce somatostatin correlated with nonuniform expression of beta-galactosidase, suggesting that promoter silencing was responsible for failure of transgene expression.


CONCLUSIONS
Retroviral gene transfer of somatostatin to canine ECs results in the production of physiologically relevant concentrations of biologically active somatostatin. Significant species differences exist in EC production of somatostatin, with promoter silencing being a potential mechanism of failure of gene expression. Gene therapy strategies using retroviral transfer of somatostatin to ECs may allow somatostatin delivery to focal areas of the vasculature.
Sarkar
R
R
Department of Surgery, University of Michigan Medical Center, Ann Arbor, USA.
Finniss
S
S
Dickinson
C J
CJ
Stanley
J C
JC
D000818:Animals; D001013:Aorta, Thoracic / Q000166:cytology / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs; D016503:Drug Delivery Systems; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D015870:Gene Expression; D005786:Gene Expression Regulation; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D008833:Microcirculation / Q000166:cytology / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011499:Protein Processing, Post-Translational; D051381:Rats; D012190:Retroviridae / Q000235:genetics; D013004:Somatostatin / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D013045:Species Specificity; D014161:Transduction, Genetic / Q000235:genetics; D014680:Veins / Q000166:cytology / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D018076:DNA, Complementary; D013004:Somatostatin; D001616:beta-Galactosidase