Bipartite Adenoviral Vector Encoding hHGF and hIL-1Ra for Improved Human Islet Transplantation

• Published in: Pharmaceutical research 2009-03, Vol.26 (3), p.587-596
• Main Authors: Panakanti, Ravikiran, Mahato, Ram I
• Format: Article
• Language: English
• Subjects: adenoviral vectors; Adenoviridae - genetics; Adenoviruses; Animals; Apoptosis; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Biomedicine; Blood Glucose - analysis; Cell Proliferation; Cell Survival; Diabetes; Diabetes Mellitus, Experimental - surgery; Diabetes Mellitus, Type 1 - surgery; Gene expression; Gene therapy; General pharmacology; Genetic Therapy; Genetic Vectors - genetics; Glucose Tolerance Test; hepatocyte growth factor; Hepatocyte Growth Factor - biosynthesis; Hepatocyte Growth Factor - genetics; hIL-1Ra; human islets; Humans; Interleukin 1 Receptor Antagonist Protein - biosynthesis; Interleukin 1 Receptor Antagonist Protein - genetics; islet transplantation; Islets of Langerhans - cytology; Islets of Langerhans - metabolism; Islets of Langerhans Transplantation; Medical Law; Medical sciences; Mice; Mice, SCID; Pancreas; Pharmaceutical sciences; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Pharmacology/Toxicology; Pharmacy; Plasmids; Research Paper; Transplants & implants
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1007/s11095-008-9777-y

Purpose Ex vivo gene therapy can improve the outcome of islet transplantation for treating type I diabetes. Hepatocyte growth factor (HGF) increases β-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-α, IL-1β and IFN-γ, 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.