Defective adenoassociated viral-mediated transfection of insulin gene by direct injection into liver parenchyma decreases blood glucose of diabetic mice.

Publication Type	Academic Article
Authors	Sugiyama A, Hattori S, Tanaka S, Isoda F, Kleopoulos S, Rosenfeld M, Kaplitt M, Sekihara H, Mobbs C
Journal	Horm Metab Res
Volume	29
Issue	12
Pagination	599-603
Date Published	12/01/1997
ISSN	0018-5043
Keywords	Dependovirus, Diabetes Mellitus, Experimental, Genetic Therapy, Insulin
Abstract	The present study assessed the feasibility of transferring the insulin gene into liver cells of diabetic individuals using a defective adenoassociated viral (AAV) vehicle. AAV offers several advantages over other viral vectors, since this vehicle can facilitate transfection in vivo without cell division and without any viral coding sequences (thus minimizing inflammation). The rat insulin gene and lacZ were each packed into a defective AAV vehicle (AAV-INS and AAV-lacZ, respectively). Successful AAV-mediated transfection and expression of lacZ into hepatocytes in primary cell culture were demonstrated by chemiluminescent assay of beta-galactosidase. Similarly, AAV-mediated transfection and expression of the insulin gene into hepatocytes was demonstrated by immunocytochemistry and reverse-transcriptase polymerase chain reaction (RT-PCR). After AAV-mediated transfection of the insulin gene into hepatocytes, glucose in the medium was significantly reduced for up to 5 days. After direct injection of AAV-INS into liver parenchyma of diabetic mice, successful transfection was demonstrated by RT-PCR, and blood glucose was significantly decreased for at least 6 days. These studies suggest that the AAV vector may be used to transfer the insulin gene into liver cells in vitro and in vivo.
DOI	10.1055/s-2007-979108
PubMed ID	9497894