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



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.



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.



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.
Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul, 133-791, South Korea.
Manbok
M
Choi
Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul, 133-791, South Korea.
Jungju
J
Oh
Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul, 133-791, South Korea. rhim@hanyang.ac.kr.
Taiyoun
T
Rhim
Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul, 133-791, South Korea. minhyung@hanyang.ac.kr.
Minhyung
M
Lee
D000818:Animals; D001921:Brain / Q000187:drug effects; D045744:Cell Line, Tumor; D003907:Dexamethasone / Q000008:administration & dosage; D004195:Disease Models, Animal; D004921:Erythropoietin / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D051547:Heme Oxygenase-1 / Q000235:genetics*; D000860:Hypoxia / Q000235:genetics*; D020244:Infarction, Middle Cerebral Artery / Q000235:genetics / Q000628:therapy; D008297:Male; D011073:Polyamines / Q000008:administration & dosage; D051381:Rats; D017207:Rats, Sprague-Dawley; D020521:Stroke / Q000235:genetics* / Q000628:therapy*; D014162:Transfection / Q000379:methods
C531249:Poly(amidoamine); D011073:Polyamines; D004921:Erythropoietin; D003907:Dexamethasone; D051547:Heme Oxygenase-1
gene delivery; heme oxygenase-1; hypoxia-inducible gene; ischemic stroke; site-specific gene therapy