Bipartite adenoviral vector encoding hHGF and hIL-1Ra for improved human islet transplantation

Pharm Res. 2009 Mar;26(3):587-96. doi: 10.1007/s11095-008-9777-y. Epub 2008 Nov 12.

Abstract

Purpose: 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.

Publication types

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

MeSH terms

  • Adenoviridae / genetics
  • Animals
  • Apoptosis
  • Blood Glucose / analysis
  • Cell Proliferation
  • Cell Survival
  • Diabetes Mellitus, Experimental / surgery
  • Diabetes Mellitus, Type 1 / surgery
  • Genetic Therapy*
  • Genetic Vectors / genetics*
  • Glucose Tolerance Test
  • Hepatocyte Growth Factor / biosynthesis*
  • Hepatocyte Growth Factor / genetics
  • Humans
  • Interleukin 1 Receptor Antagonist Protein / biosynthesis*
  • Interleukin 1 Receptor Antagonist Protein / genetics
  • Islets of Langerhans / cytology*
  • Islets of Langerhans / metabolism
  • Islets of Langerhans Transplantation*
  • Mice
  • Mice, SCID
  • Plasmids

Substances

  • Blood Glucose
  • HGF protein, human
  • Interleukin 1 Receptor Antagonist Protein
  • Hepatocyte Growth Factor