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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
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Akt/protein kinase B activation by adenovirus vectors contributes to NFkappaB-dependent CXCL10 expression

Qiang Liu1, Lindsay R White, Sharon A Clark

  • 1Department of Medicine, University of Calgary, Calgary, Alberta T2N 4N1, Canada.

Journal of Virology
|November 12, 2005
PubMed

Insights

Adenovirus vectors trigger immune responses via the Akt pathway, which activates NFkappaB and CXCL10 expression in kidney cells. Targeting this pathway can improve gene therapy safety and efficacy.

Area of Science:

  • Gene Therapy
  • Immunology
  • Molecular Biology

Background:

  • Adenovirus (Ad) vectors are crucial for gene therapy but face innate immune system barriers.
  • Serotype 5 Ad vectors induce CXCL10 (IP-10) chemokine expression in kidney cells, mediated by NFkappaB.
  • RGD-alpha(V) integrin interactions are implicated in Ad vector-induced inflammatory gene expression.

Purpose of the Study:

  • To investigate the role of the phosphoinositide-3-OH kinase/Akt pathway in Ad vector-induced immune responses.
  • To elucidate the mechanism by which Ad vectors activate NFkappaB and CXCL10 expression.
  • To identify strategies for improving Ad vector-based gene therapy efficacy and safety.

Main Methods:

  • Comparing AdLuc and RGD-deleted AdL.PB vectors for CXCL10 activation.
  • Assessing Akt activation kinetics and dose-dependency post-transduction.
  • Utilizing pharmacological inhibitors (Wortmannin, Ly294002) and dominant-negative Akt (AktAAA) to block the Akt pathway.
  • Employing luciferase reporter assays to evaluate promoter activation.

Main Results:

  • RGD-deleted AdL.PB vectors showed reduced CXCL10 activation compared to AdLuc.
  • Akt activation was rapid, dose-dependent, and dependent on capsid-alpha(V) integrin interactions.
  • Inhibition of the Akt pathway decreased Ad vector-induced CXCL10 mRNA expression.
  • Akt activation was found to mediate NFkappaB-dependent transcriptional activation of CXCL10.

Conclusions:

  • The Akt pathway plays a significant role in Ad vector-mediated activation of NFkappaB and CXCL10 expression.
  • Understanding this mechanism is key to developing safer and more effective Ad vector gene therapies.
  • Targeting the Akt/NFkappaB signaling axis could mitigate detrimental immune responses to Ad vectors.

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