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Pharmaceutical Research

, Volume 33, Issue 9, pp 2250–2258 | Cite as

Delivery of Hypoxia-Inducible Heme Oxygenase-1 Gene for Site-Specific Gene Therapy in the Ischemic Stroke Animal Model

  • Manbok Choi
  • Jungju Oh
  • Taiyoun RhimEmail author
  • Minhyung LeeEmail author
Research Paper

ABSTRACT

Purpose

To reduce side effects due to non-specific expression, the heme oxygenase-1 (HO-1) gene under control of a hypoxia-inducible erythropoietin (Epo) enhancer (pEpo-SV-HO-1) was developed for site-specific gene therapy of ischemic stroke.

Methods

pEpo-SV-HO-1 was constructed by insertion of the Epo enhancer into pSV-HO-1. Dexamethasone-conjugated polyamidoamine (PAMAM-Dexa) was used as a gene carrier. In vitro transfection assays were performed in the Neuro2A cells. In vivo efficacy of pEpo-SV-HO-1 was evaluated in the transient middle cerebral artery occlusion (MCAO) model.

Results

In vitro transfection assay with the PAMAM-Dexa/pEpo-SV-HO-1 complex showed that pEpo-SV-HO-1 had higher HO-1 gene expression than pSV-HO-1 under hypoxia. In addition, pEpo-SV-HO-1 reduced the level of apoptosis more efficiently than pSV-HO-1 in Neuro2A cells under hypoxia. For in vivo evaluation, the PAMAM-Dexa/pEpo-SV-HO-1 complex was injected into the ischemic brain of the transient MCAO model. pEpo-SV-HO-1 increased HO-1 expression and reduced the number of apoptotic cells in the ischemic brain, compared with the pSV-HO-1 injection group. As a result, the infarct volume was more efficiently decreased by pEpo-SV-HO-1 than by pSV-HO-1.

Conclusions

pEpo-SV-HO-1 induced HO-1 gene expression and therapeutic effect in the ischemic brain. Therefore, pEpo-SV-HO-1 may be useful for site-specific gene therapy of ischemic stroke.

KEY WORDS

gene delivery heme oxygenase-1 hypoxia-inducible gene ischemic stroke site-specific gene therapy 

ABBREVIATIONS

(DMSO)

Dimethylsulfoxide

(Epo)

Erythropoietin

(HIF-1)

Hypoxia-inducible factor-1

(HO-1)

Heme oxygenase-1

(HREs)

Hypoxia response elements

(LDH)

Lactate dehydrogenase

(MCAO)

Middle cerebral artery occlusion

(NMDA)

N-methyl-D-aspartic acid

(PAMAM G2)

Polyamidoamine generation 2

(pDNA)

Plasmid DNA

(ROS)

Reactive oxygen species

(SD)

Sprague Dawley

(SV40)

Simian virus 40

(TTC)

2, 3, 5-triphenyl tetrazolium chloride

(VEGF)

Vascular endothelial growth factor

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

This work was supported by a grant from the National Research Foundation of Korea, funded by the Ministry of Science, ICT and Future Planning (NRF-2013R1A1A2059236).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Bioengineering, College of EngineeringHanyang UniversitySeongdong-guSouth Korea

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