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

R. Panakanti, Ram I Mahato

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)587-596
Number of pages10
JournalPharmaceutical Research
Volume26
Issue number3
DOIs
StatePublished - Mar 1 2009

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Islets of Langerhans Transplantation
Caspase 3
Genes
Insulin
Gene Expression
Gene therapy
Inbred NOD Mouse
Interleukin-1 Receptors
Hepatocyte Growth Factor
SCID Mice
Cloning
Cell proliferation
Streptozocin
Medical problems
Protein Sorting Signals
Type 1 Diabetes Mellitus
Interleukin-1
Islets of Langerhans
Gene expression
Genetic Therapy

Keywords

  • Adenoviral vectors
  • HIL-1Ra
  • Hepatocyte growth factor
  • Human islets
  • Islet transplantation

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Organic Chemistry
  • Molecular Medicine
  • Pharmacology (medical)
  • Biotechnology
  • Pharmacology

Cite this

Bipartite adenoviral vector encoding hHGF and hIL-1Ra for improved human islet transplantation. / Panakanti, R.; Mahato, Ram I.

In: Pharmaceutical Research, Vol. 26, No. 3, 01.03.2009, p. 587-596.

Research output: Contribution to journalArticle

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