Coexpression of Vascular Endothelial Growth Factor and interleukin-1 Receptor Antagonist for Improved Human Islet Survival and Function

Mol Pharm. Mar-Apr 2007;4(2):199-207. doi: 10.1021/mp060091s. Epub 2007 Jan 26.

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-1alpha (EF-1 alpha) 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-1beta, TNFalpha, and IFNgamma. In conclusion, our results indicated that coexpression of growth factor and antiapoptic genes can improve islet survival and function.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Caspases / metabolism
  • Cell Death
  • Cells, Cultured
  • Cytokines / pharmacology
  • Gene Expression Regulation*
  • Genetic Vectors / genetics*
  • Humans
  • Interleukin 1 Receptor Antagonist Protein / metabolism*
  • Islets of Langerhans / physiology*
  • Nitric Oxide / metabolism
  • Time Factors
  • Transfection
  • Vascular Endothelial Growth Factor A / metabolism*

Substances

  • Cytokines
  • Interleukin 1 Receptor Antagonist Protein
  • Vascular Endothelial Growth Factor A
  • Nitric Oxide
  • Caspases