Bipartite adenoviral vector encoding hHGF and hIL-1Ra for improved human islet transplantation.
OBJECTIVE
Ex vivo gene therapy can improve the outcome of islet transplantation for treating type I diabetes. Hepatocyte growth factor (HGF) increases beta-cell proliferation and promotes revascularization of islets, while interleukin-1 receptor antagonist (hIL-1Ra) inhibits islet cell apoptosis.


METHODS
We constructed Adv-hHGF-hIL-1Ra by cloning hHGF and hIL-1Ra coding sequences and polyA signal under separate CMV promoters in Adenoquick plasmid.


RESULTS
There was dose and time dependent expression of these genes after transduction of Adv-hHGF-hIL-1Ra into human islets. Compared to un-transduced islets, hHGF and hIL-1Ra gene expression at protein levels was more than 60 and 40 times higher at 1,000 MOI, respectively. Transduced islets were viable after incubation with the cocktail of TNF-alpha, IL-1beta and IFN-gamma, as evidenced by insulin release in response to glucose concentration. Co-expression of hHGF and hIL-1Ra led to significant decrease in caspase-3 induced by the cytokines. Compared to un-transduced islets, transduction of islets with Adv-hHGF-hIL-1Ra at 1,000 MOI prior to transplantation under the kidney capsules of streptozotocin-induced-diabetic NOD-SCID mice reduced blood glucose levels, and increased serum insulin and c-peptide levels.


CONCLUSIONS
Transduction of islets with Adv-hHGF-hIL-1Ra efficiently expresses both growth factor and antiapoptotic genes, decreases caspase-3 and improves the outcome of islet transplantation.
Panakanti
Ravikiran
R
Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, 19 S Manassas, RM 224, Memphis, Tennessee, 38103-3308, USA.
Mahato
Ram I
RI
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D001786:Blood Glucose / Q000032:analysis; D049109:Cell Proliferation; D002470:Cell Survival; D003921:Diabetes Mellitus, Experimental / Q000601:surgery; D003922:Diabetes Mellitus, Type 1 / Q000601:surgery; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D005951:Glucose Tolerance Test; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein / Q000096:biosynthesis* / Q000235:genetics; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D016381:Islets of Langerhans Transplantation; D051379:Mice; D016513:Mice, SCID; D010957:Plasmids
D001786:Blood Glucose; C066855:HGF protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D017228:Hepatocyte Growth Factor