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Translational Stroke Research

, Volume 2, Issue 1, pp 92–100 | Cite as

An Insult-Inducible Vector System Activated by Hypoxia and Oxidative Stress for Neuronal Gene Therapy

  • Michelle Y. ChengEmail author
  • I-Ping Lee
  • Michael Jin
  • Guohua Sun
  • Heng Zhao
  • Gary K. Steinberg
  • Robert M. Sapolsky
Article

Abstract

Gene therapy has demonstrated the protective potential of a variety of genes against stroke. However, conventional gene therapy vectors are limited due to the inability to temporally control their expression, which can sometimes lead to deleterious side effects. Thus, an inducible vector that can be temporally controlled and activated by the insult itself would be advantageous. Using hypoxia responsive elements (HRE) and antioxidant responsive elements (ARE), we have constructed an insult-inducible vector activated by hypoxia and reactive oxygen species (ROS). In COS7 cells, the inducible ARE–HRE-luciferase vectors are highly activated by oxygen deprivation, hydrogen peroxide treatment, and the ROS-induced transcription factor NF-E2-related factor 2 (Nrf2). Using a defective herpes virus, the neuroprotective potential of this inducible vector was tested by over-expressing the transcription factor Nrf2. In primary cortical cultures, expression of the inducible ARE–HRE–Nrf2 protects against oxygen glucose deprivation, similar to that afforded by the constitutively expressed Nrf2. This ARE + HRE vector system is advantageous in that it allows the expression of a transgene to be activated not only during hypoxia but also maintained after reperfusion, thus prolonging the transgene expression during an ischemic insult. This insult-inducible vector system will be a valuable gene therapy tool for activating therapeutic/protective genes in cerebrovascular diseases.

Keywords

Insult-inducible Gene therapy Hypoxia HIF1 Nrf2 Reactive oxygen species 

Notes

Acknowledgments

We would like to thank Dr. Jewed Alam and Dr. Howard Federoff for kindly providing the pEF-Nrf2 plasmid and the replication defective HSV plasmid, respectively. This work was supported by the NIH-Center for Cerebrovascular Disease no. 2P01 NS37520. MYC was supported by the American Heart Association (no. 0525025Y).

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Michelle Y. Cheng
    • 1
    • 2
    Email author
  • I-Ping Lee
    • 1
  • Michael Jin
    • 1
  • Guohua Sun
    • 2
  • Heng Zhao
    • 2
  • Gary K. Steinberg
    • 2
  • Robert M. Sapolsky
    • 1
    • 2
  1. 1.Department of BiologyStanford UniversityStanfordUSA
  2. 2.Department of NeurosurgeryStanford University School of MedicineStanfordUSA

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