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Nitric Oxide pp 153-168 | Cite as

β Cell Protection by Inhibition of iNOS Through Lentiviral Vector-Based Strategies

  • Sean O. HynesEmail author
  • Cillian McCabe
  • Timothy O’Brien
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 704)

Abstract

Cytoprotective gene transfer to pancreatic islet β cell s may prove useful in preventing their destruction and prolonging islet graft survival after transplantation in patients with type 1 diabetes mellitus. A host of therapeutically relevant transgenes may potentially be incorporated into an appropriate gene delivery vehicle and used for islet modification. To examine this, we utilised a robust model of cytokine-induced β cell pathophysiology. Using this model, it is clear that antioxidant gene transfer confers no cytoprotective benefit. In contrast, we demonstrated that gene-based approaches to inhibit the activation of NF-κB following cytokine exposure harbours therapeutic utility in preserving islet β cell viability in the face of cytokine toxicity. We identified that NF-κB-dependent induction of iNOS is a critical determinant of β cell fate following cytokine exposure. Having identified the pivotal role of iNOS activation in cytokine-induced β cell pathophysiology, lentiviral vectors may be used to efficiently deliver small interfering RNA molecules to confer efficient iNOS gene silencing. We have shown that lentiviral vector-based shRNA delivery holds significant promise in preserving β cell viability following cytotoxic cytokine exposure.

Key words

Diabetes mellitus pancreatic islet cells iNOS small interfering RNAs NF-κB gene silencing gene therapy 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Sean O. Hynes
    • 1
    Email author
  • Cillian McCabe
    • 1
  • Timothy O’Brien
    • 1
  1. 1.Regenerative Medicine Institute, National University of IrelandGalwayIreland

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