Incorporation of calcium phosphate enhances recombinant adeno-associated virus-mediated gene therapy in diabetic mice
Journal
Journal of Gene Medicine
Journal Volume
5
Journal Issue
5
Pages
417-424
Date Issued
2003
Author(s)
YA-WUN YANG
Chao C.-K.
Abstract
Background: Increased efficiency of transgene expression is desired for virus-mediated gene delivery. In the present study, we examined the effect of calcium phosphate (CaPi) on recombinant adenoassociated virus (rAAV)-mediated insulin therapy in diabetic animals. Methods: The rAAV vector, rAAV.PEPCK.Ins.EGFP, containing the human insulin gene under control of the phosphoenolpyruvate carboxykinase (PEPCK) promoter and the enhanced green fluorescence protein (EGFP) gene driven by the cytomegalovirus (CMV) IE promoter, was employed in this study. C57BL/6J mice were made diabetic with streptozotocin (STZ), followed by injection into the livers with either rAAV alone, or noncovalent complexes with calcium phosphate. Body weight and blood glucose levels of the animals were routinely monitored after 6 h fasting. Secretion of human insulin in the rAAV-transduced animals was determined by radioimmunoassay (RIA). Expression of human insulin in the livers of the animals was detected by immunohistochemical staining. Results: Compared with the STZ-treated control mice, administration of rAAV containing the human insulin gene significantly decreased blood glucose levels and maintained body weight of the diabetic animals. Complexation of rAAV with calcium phosphate enhanced the hypoglycemic effect of rAAV-mediated gene transfer. Results obtained from both RIA and immunohistochemical staining demonstrated that incorporation of calcium phosphate enhanced rAAV-mediated gene transfer in vivo, leading to higher expression and secretion of human insulin. Conclusions: Administration of rAAV harboring the human insulin gene into livers of the STZ-diabetic mice improved blood glucose levels, maintained body weight of the diabetic animals, and resulted in human insulin secretion. Complexation of rAAV with calcium phosphate significantly potentiated the efficiency of rAAV-mediated diabetic gene therapy. Copyright ? 2002 John Wiley & Sons, Ltd.
Subjects
Calcium phosphate; Diabetes mellitus; Gene therapy; Human insulin; PEPCK; Recombinant adeno-associated virus
SDGs
Other Subjects
calcium phosphate; glucose; green fluorescent protein; insulin; parvovirus vector; phosphoenolpyruvate carboxykinase (pyrophosphate); streptozocin; alpha tricalcium phosphate; alpha-tricalcium phosphate; calcium phosphate monobasic; dicalcium phosphate anhydrous; insulin; phosphoenolpyruvate carboxykinase (GTP); tetracalcium phosphate; animal cell; animal experiment; animal model; article; blood glucose monitoring; body weight; controlled study; Cytomegalovirus; diabetes mellitus; disease model; gene transfer; glucose blood level; immunohistochemistry; in vivo study; insulin release; male; mouse; mouse strain; nonhuman; priority journal; promoter region; radioimmunoassay; streptozocin diabetes; viral gene delivery system; viral gene therapy; virus recombinant; Adeno associated virus; animal; C57BL mouse; cell line; cell strain COS1; experimental diabetes mellitus; gene therapy; gene vector; genetics; human; liver; metabolism; methodology; time; Animals; Blood Glucose; Body Weight; Calcium Phosphates; Cell Line, Transformed; COS Cells; Cytomegalovirus; Dependovirus; Diabetes Mellitus, Experimental; Gene Therapy; Genetic Vectors; Humans; Immunohistochemistry; Insulin; Liver; Male; Mice; Mice, Inbred C57BL; Phosphoenolpyruvate Carboxykinase (GTP); Promoter Regions (Genetics); Radioimmunoassay; Time Factors
Type
journal article