Adeno-associated viral delivery of a metabolically regulated insulin transgene to hepatocytes.
Transduction with a liver specific, metabolically responsive insulin transgene produces near-normal blood sugars in STZ-diabetic rats. To overcome the limited duration of hepatic transgene expression induced by E1A-deleted adenoviral vectors, we evaluated recombinant adeno-associated virus (rAAV2) for cell type specificity and glucose responsiveness in vitro. Co-infection of AAV2 containing the glucose responsive, liver-specific (GlRE)(3)BP-1 promoter with an empty adenovirus enhanced transduction efficiency, and shortened the duration of transgene expression in HepG2 hepatoma cells, but not primary hepatocytes. However, in the context of rAAV2, (GlRE)(3)BP-1 promoter activity remained confined to cells of hepatocyte lineage, and retained glucose responsiveness. While isolated infection with an insulin expressing rAAV2 failed to attenuate blood sugars in diabetic mice, adenoviral co-administration with the same rAAV2 induced transient, near-normal random blood sugars in a diabetic animal. We conclude that rAAV2 can induce metabolically responsive insulin secretion from hepatocytes in vitro and in vivo. However, alternative AAV serotypes will likely be required to efficiently deliver therapeutic genes to the liver for the treatment of diabetes mellitus.
Kozlowski
Miroslaw
M
Department of Orthopedics, Veterans Affairs Medical Center and Emory University School of Medicine, Atlanta, GA 30033, USA.
Olson
Darin E
DE
Rubin
Janet
J
Lyszkowicz
Dorota
D
Campbell
Adam
A
Thulé
Peter M
PM
D000818:Animals; D002478:Cells, Cultured; D000229:Dependovirus / Q000187:drug effects / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D005947:Glucose / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006943:Hyperglycemia / Q000139:chemically induced / Q000473:pathology; D007328:Insulin / Q000235:genetics*; D008297:Male; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013311:Streptozocin; D013997:Time Factors; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics*
D007328:Insulin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D013311:Streptozocin; D005947:Glucose