Controlled nonviral gene delivery and expression using stable neural stem cell line transfected with a hypoxia-inducible gene expression system
- Authors
- Liu, Meng-Lu; Oh, Jin Soo; An, Sung Su; Pennant, William A.; Kim, Hyo Jin; Gwak, So-Jung; Yoon, Do Heum; Kim, Keung Nyun; Lee, Minhyung; Ha, Yoon
- Issue Date
- Dec-2010
- Publisher
- WILEY
- Keywords
- controllable gene therapy; erythropoietin enhancer; hypoxia; nonviral gene delivery; neural stem cell; spinal cord injury
- Citation
- JOURNAL OF GENE MEDICINE, v.12, no.12, pp.990 - 1001
- Indexed
- SCIE
SCOPUS
- Journal Title
- JOURNAL OF GENE MEDICINE
- Volume
- 12
- Number
- 12
- Start Page
- 990
- End Page
- 1001
- URI
- https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/173372
- DOI
- 10.1002/jgm.1527
- ISSN
- 1099-498X
- Abstract
- Background 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. Copyright (c) 2010 John Wiley & Sons, Ltd.
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