DEFECTIVE ADENOASSOCIATED VIRAL-MEDIATED TRANSFECTION OF INSULIN GENEBY DIRECT-INJECTION INTO LIVER PARENCHYMA DECREASES BLOOD-GLUCOSE OF DIABETIC MICE

The present study assessed the feasibility of transferring the insulin gene into liver cells of diabetic individuals using a defective adeno associated viral (AAV) vehicle. AAV offers several advantages over oth er viral vectors, since this vehicle can facilitate transfection in vi vo without cell division and without any viral coding sequences (thus minimizing inflammation). The rat insulin gene and lacZ were each pack ed into a defective AAV vehicle (AAV-INS and AAV-lacZ, respectively). Successful AAV-mediated transfection and expression of lacZ into hepat ocytes in primary cell culture were demonstrated by chemiluminescent a ssay of beta-galactosidase. Similarly, AAV-mediated transfection and e xpression of the insulin gene into hepatocytes was demonstrated by imm unocytochemistry and reverse-transcriptase polymerase chain reaction ( RT-PCR), After AAV-mediated transfection of the insulin gene into hepa tocytes, glucose in the medium was significantly reduced for up to 5 d ays. After direct injection of AAV-INS into liver parenchyma of diabet ic mice, successful transfection was demonstrated by RT-PCR, and blood glucose was significantly decreased for at least 6 days. These studie s suggest that the AAV vector may be used to transfer the insulin gene into liver cells in vitro and in vivo.