Liver-directed gene therapy of diabetic rats using an HVJ-E vector containing EBV plasmids expressing insulin and GLUT 2 transporter.
Insulin gene therapy in clinical medicine is currently hampered by the inability to regulate insulin secretion in a physiological manner, the inefficiency with which the gene is delivered, and the short duration of gene expression. To address these issues, we injected the liver of streptozotocin-induced diabetic rats with hemagglutinating virus of Japan-envelope (HVJ-E) vectors containing Epstein-Barr virus (EBV) plasmids encoding the genes for insulin and the GLUT 2 transporter. Efficient delivery of the genes was achieved with the HVJ-E vector, and the use of the EBV replicon vector led to prolonged hepatic gene expression. Blood glucose levels were normalized for at least 3 weeks as a result of the gene therapy. Cotransfection of GLUT 2 with insulin permitted the diabetic rats to regulate their blood glucose levels upon exogenous glucose loading in a physiologically appropriate manner and improved postprandial glucose levels. Moreover, cotransfection with insulin and GLUT 2 genes led to in vitro glucose-stimulated insulin secretion that involved the closure of K(ATP) channels. The present study represents a new way to efficiently deliver insulin gene in vivo that is regulated by ambient glucose level with prolonged gene expression. This may provide a basis to overcome limitations of insulin gene therapy in humans.
Kim
Y D
YD
Department of Internal Medicine & Institute for Medical Sciences, Keimyung University School of Medicine, Daegu, South Korea.
Park
K-G
KG
Morishita
R
R
Kaneda
Y
Y
Kim
S-Y
SY
Song
D-K
DK
Kim
H-S
HS
Nam
C-W
CW
Lee
H C
HC
Lee
K-U
KU
Park
J-Y
JY
Kim
B-W
BW
Kim
J-G
JG
Lee
I-K
IK
D000818:Animals; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism / Q000628:therapy*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D051273:Glucose Transporter Type 2 / Q000235:genetics* / Q000378:metabolism; D004854:Herpesvirus 4, Human / Q000235:genetics; D007328:Insulin / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008297:Male; D015221:Potassium Channels / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D029082:Sendai virus / Q000235:genetics; D013997:Time Factors; D014161:Transduction, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics
D051273:Glucose Transporter Type 2; D007328:Insulin; D015221:Potassium Channels; D014759:Viral Envelope Proteins