PURPOSE: The purpose of this study was to examine glucose- and metabolically modulation of insulin secretion by rAAV-mediated gene delivery in vitro and in vivo. MATERIALS AND METHODS: A recombinant adeno-associated virus vector (rAAV) containing a furin-mutated human insulin gene, driven by the rat insulin I promoter, was used in this study. Glucose-responsive secretion of human insulin was determined by treating rAAV-transduced Huh7 human hepatoma cells with varying concentrations of glucose, with or without insulin secretagogues. Glucose- and metabolically modulated secretion of human insulin in the streptozotocin (STZ)-induced diabetic mice was assessed by intrahepatic administration of rAAV-polyethylenimine (PEI) complexes, followed by intraperitoneal glucose tolerance test (IPGTT), with or without theophylline. RESULTS: Glucose- and metabolically controlled human insulin secretion was obtained in the rAAV-transduced Huh7 cells. Treatment of STZ-induced diabetic animals with rAAV-polyethylenimine (rAAV-PEI) complexes resulted in production of human insulin and amelioration of hyperglycemia. Co-administration of glucose and theophylline in these animals augmented the secretion of human insulin, demonstrating metabolic modulation of insulin secretion in vivo. Immunohistochemical examination of the liver sections of rAAV-treated mice confirmed the production of human insulin. CONCLUSIONS: Glucose- and metabolically controlled hepatic insulin gene therapy was obtained both in vitro and in vivo.
PURPOSE: The purpose of this study was to examine glucose- and metabolically modulation of insulin secretion by rAAV-mediated gene delivery in vitro and in vivo. MATERIALS AND METHODS: A recombinant adeno-associated virus vector (rAAV) containing a furin-mutated humaninsulin gene, driven by the ratinsulin I promoter, was used in this study. Glucose-responsive secretion of human insulin was determined by treating rAAV-transduced Huh7 humanhepatoma cells with varying concentrations of glucose, with or without insulin secretagogues. Glucose- and metabolically modulated secretion of humaninsulin in the streptozotocin (STZ)-induced diabeticmice was assessed by intrahepatic administration of rAAV-polyethylenimine (PEI) complexes, followed by intraperitoneal glucose tolerance test (IPGTT), with or without theophylline. RESULTS:Glucose- and metabolically controlled humaninsulin secretion was obtained in the rAAV-transduced Huh7 cells. Treatment of STZ-induced diabetic animals with rAAV-polyethylenimine (rAAV-PEI) complexes resulted in production of humaninsulin and amelioration of hyperglycemia. Co-administration of glucose and theophylline in these animals augmented the secretion of humaninsulin, demonstrating metabolic modulation of insulin secretion in vivo. Immunohistochemical examination of the liver sections of rAAV-treated mice confirmed the production of humaninsulin. CONCLUSIONS:Glucose- and metabolically controlled hepatic insulin gene therapy was obtained both in vitro and in vivo.
Authors: M Tiedge; M Elsner; N H McClenaghan; H J Hedrich; D Grube; J Klempnauer; S Lenzen Journal: Hum Gene Ther Date: 2000-02-10 Impact factor: 5.695
Authors: Guo Jun Liu; Ann M Simpson; M Anne Swan; Chang Tao; Bernard E Tuch; Russell M Crawford; Aleksandar Jovanovic; Donald K Martin Journal: FASEB J Date: 2003-07-18 Impact factor: 5.191