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Horm Metab Res 1997; 29(12): 599-603
DOI: 10.1055/s-2007-979108
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© Georg Thieme Verlag Stuttgart · New York

Defective Adenoassociated Viral-Mediated Transfection of Insulin Gene by Direct Injection into Liver Parenchyma Decrease Blood Glucose of Diabetic Mice

A. Sugiyama1 , 2 , S. Hattori3 , S. Tanaka2 , F. Isoda3 , S. Kleopoulos2 , M. Rosenfeld4 , M. Kaplitt5 , H. Sekihara2 , C. Mobbs1 , 6
  • 1Fishberg Center for Neurobiology and Department of Geriatrics, Mt. Sinai School of Medicine, New York, USA
  • 2The Third Department of Internal Medicine, Yokohama City University School of Medicine, Yokohama, Japan
  • 3Health Sciences Research Institute, Yokohama, Japan
  • 4Dept. of Neurology and the Cotzias Laboratory of Neuro-Oncology, Memorial Sloan-Kettering Cancer Center, New York, USA
  • 5Dept. of Neurosurgery, New York Hospital, Cornell Medical School, and Rockefeller University, New York, USA
  • 6Pathology & Diagnostic Laboratory, Veterans Administration Medical Center, New York, USA
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Publication History

1996

1997

Publication Date:
23 April 2007 (online)

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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.

Key words

Gene Therapy - AAV - Gene Transfer - Defective Vector

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