Increased glucose disposal induced by adenovirus-mediated transfer of glucokinase to skeletal muscle in vivo

In non-insulin-dependent diabetes mellitus, insulin-stimulated glucose uptake is impaired in muscle, contributing in a major way to development of hyperglycemia, We previously showed that expression of the glucose phosphorylating enzyme glucokinase (GK) in cultured human myocytes improved glucose storage and disposal, suggesting that GK delivery to muscle in situ could potentially enhance glucose clearance. Here we have tested this idea directly by intramuscular delivery of an adenovirus containing the liver GK cDNA (AdCMV-GKL) into one hind limb, We injected an adenovirus containing the beta-galactosidase gene (AdCMV-lacZ) into the hind limb of newborn rats. beta-Galactosidase activity was localized in muscle for as long as 1 month after delivery, with a large percentage of fibers staining positive in the gastrocnemius, Using the same approach with AdCMV-GKL, GK protein content was increased from zero to 50-400% of the GK in normal liver sample, and total glucose phosphorylating activity was increased in GK-expressing muscles relative to the counterpart uninfected muscle, Expression of GK in muscle improved glucose tolerance rather than changing basal glycemic control. Glucose levels were reduced by similar to 35% 10 min after administration of a glucose bolus to fed animals treated with AdCMV-GKL relative to AdCMV-lacZ-treated controls. The enhanced rate of glucose clearance was reflected in increases in muscle 2-deoxy glucose uptake and blood lactate levels. We conclude that restricted expression of GK in muscle leads to an enhanced capacity for muscle glucose disposal and whole body glucose tolerance under conditions of maximal glucose-insulin stimulation, suggesting that under these conditions glucose phosphorylation becomes rate-limiting. Our findings also show that gene delivery to a fraction of the whole body is sufficient to improve glucose disposal, providing a rationale for the development of new therapeutic strategies for treatment of diabetes.