Controlled nonviral gene delivery and expression using stable neural stem cell line transfected with a hypoxia-inducible gene expression system

Meng Lu Liu, Jin Soo Oh, Sung Su An, William A. Pennant, Hyo Jin Kim, So Jung Gwak, Do Heum Yoon, Keung Nyun Kim, Minhyung Lee, Yoon Ha

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Nonviral ex vivo local gene therapy systems consisting of regulated gene expression vectors and cellular delivery platforms represent a novel strategy for tissue repair and regeneration. We introduced a hypoxia-regulated plasmid-based system into mouse neural stem cells (NSCs) as an efficient gene expression and delivery platform for rapid, robust and persistent hypoxic/ischemic-regulated gene expression in the spinal cord.Methods: A synthetic hypoxia-responsive erythropoietin (Epo) enhancer, the SV40 minimal promoter and the luciferase (Luc) reporter gene were incorporated in a DsRed-expressing double-promoter plasmid for cell lipofection and Zeocin-selection to establish a hypoxia-regulated stable NSC line (NSC-Epo-SV-Luc). A nonhypoxia-regulated stable NSC line (NSC-SV-Luc) was also established as a control.Results: Under the transcriptional regulation of the Epo enhancer, in vitro luciferase expression in NSC-Epo-SV-Luc, but not in NSC-SV-Luc, was sensitively augmented according to the strength and duration of the hypoxic stimulus and was quickly down-regulated to a low basal level after reoxygenation of the hypoxic cells. Furthermore, deoxygenation of the reoxygenated cells clearly enhanced the luciferase activity again. After transplantation into a rat spinal cord injury (SCI) model, only NSC-Epo-SV-Luc showed ischemic injury-specific luciferase expression Notably, the engineered NSC lines kept the neural differentiation potential and retained the hypoxia-regulated luciferase expression after differentiation.Conclusions: We propose that NSCs engineered with the Epo-SV-therapeutic gene will be valuable for developing a controllable stem cell-mediated nonviral gene therapy for SCI or other central nervous system diseases accompanied with chronic or episodic hypoxic/ischemic stresses.

Original languageEnglish (US)
Pages (from-to)990-1001
Number of pages12
JournalJournal of Gene Medicine
Volume12
Issue number12
DOIs
StatePublished - Dec 1 2010

Keywords

  • Controllable gene therapy
  • Erythropoietin enhancer
  • Hypoxia
  • Neural stem cell
  • Nonviral gene delivery
  • Spinal cord injury

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Drug Discovery
  • Genetics(clinical)

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  • Cite this

    Liu, M. L., Oh, J. S., An, S. S., Pennant, W. A., Kim, H. J., Gwak, S. J., Yoon, D. H., Kim, K. N., Lee, M., & Ha, Y. (2010). Controlled nonviral gene delivery and expression using stable neural stem cell line transfected with a hypoxia-inducible gene expression system. Journal of Gene Medicine, 12(12), 990-1001. https://doi.org/10.1002/jgm.1527