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

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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.

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Acknowledgements

We would like to thank the National Institute of Health (NIH) for financial support (RO1 DK69968).

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, 19 S Manassas, RM 224, Memphis, Tennessee, 38103-3308, USA

    Ravikiran Panakanti & Ram I. Mahato

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  1. Ravikiran Panakanti
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  2. Ram I. Mahato
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Correspondence to Ram I. Mahato.

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Panakanti, R., Mahato, R.I. Bipartite Adenoviral Vector Encoding hHGF and hIL-1Ra for Improved Human Islet Transplantation. Pharm Res 26, 587–596 (2009). https://doi.org/10.1007/s11095-008-9777-y

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  • DOI: https://doi.org/10.1007/s11095-008-9777-y

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