Abstract
Ex vivo gene therapy approaches can improve the outcome of islet transplantation for treating type I diabetes. We have recently shown improvement in islet survival and function following ex vivo infection of islets with a mixture of adenoviral vectors encoding human vascular endothelial growth factor (Adv-hVEGF) and human interleukin-1 receptor antagonist (Adv-hIL-1Ra). In this study, we constructed a bicistronic vector encoding these two genes (phVEGF-hIL-1Ra) by cloning hIL-1Ra under the cytomegalovirus (CMV) promoter and hVEGF under the elongation factor-1α (EF-1 α) promoter in pBudCE4.1 vector. There was dose and time dependent expression of hVEGF and hIL-1Ra at both mRNA and protein levels after transfection with human islets. Transfected islets were viable, as evidenced by insulin release upon glucose challenge. Coexpression of hVEGF and hIL-1Ra suppressed nitric oxide production, total caspases, apoptosis, and necrosis in the presence of inflammatory cytokine cocktail consisting of IL-1β, TNFα, and IFNγ. In conclusion, our results indicated that coexpression of growth factor and antiapoptic genes can improve islet survival and function.
Original language | English (US) |
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Pages (from-to) | 199-207 |
Number of pages | 9 |
Journal | Molecular Pharmaceutics |
Volume | 4 |
Issue number | 2 |
DOIs | |
State | Published - Mar 2007 |
Externally published | Yes |
Keywords
- Bicistronic plasmid
- Human islet
- Insulin release
- Real-time RT-PCR
- hIL-1Ra
- hVEGF
ASJC Scopus subject areas
- Molecular Medicine
- Pharmaceutical Science
- Drug Discovery