Adenovirus-mediated gene transfer of heparin-binding epidermal growth factor-like growth factor enhances neurogenesis and angiogenesis after focal cerebral ischemia in rats.
OBJECTIVE
Recent studies have demonstrated that neurotrophic factors promote neurogenesis after cerebral ischemia. However, it remains unknown whether administration of genes encoding those factors could promote neural regeneration in the striatum and functional recovery. Here, we examined the efficacy of intraventricular injection of a recombinant adenovirus-expressing heparin-binding epidermal growth factor-like growth factor (HB-EGF) on neurogenesis, angiogenesis, and functional outcome after focal cerebral ischemia.


METHODS
Transient focal ischemia was induced by middle cerebral artery occlusion (MCAO) for 80 minutes with a nylon filament in Wistar rats. Three days after MCAO, either adenovirus-expressing HB-EGF (Ad-HB-EGF) or Ad-LacZ, the control vector, was injected into the lateral ventricle on the ischemic side. Bromodeoxyuridine (BrdU) was injected intraperitoneally twice daily on the sixth and seventh days. On the eighth or 28th day after MCAO, we evaluated infarct volume, neurogenesis, and angiogenesis histologically. Neurological outcome was serially evaluated by the rotarod test after MCAO.


RESULTS
There was no significant difference in infarct volume between the 2 groups. Treatment with Ad-HB-EGF significantly increased the number of BrdU-positive cells in the subventricular zone on the 8th day. In addition, on the 28th day, BrdU-positive cells differentiated into mature neurons in the striatum on the ischemic side but seldom the cells given Ad-LacZ. Enhancement of angiogenesis at the peri-infarct striatum was also observed on the eighth day in Ad-HB-EGF-treated rats. Treatment with Ad-HB-EGF significantly enhanced functional recovery after MCAO.


CONCLUSIONS
Our data suggest that gene therapy using Ad-HB-EGF contributes to functional recovery after ischemic stroke by promoting neurogenesis and angiogenesis.
Sugiura
Shiro
S
Division of Strokology, Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Japan. siro@medone.med.osaka-u.ac.jp
Kitagawa
Kazuo
K
Tanaka
Shigeru
S
Todo
Kenichi
K
Omura-Matsuoka
Emi
E
Sasaki
Tsutomu
T
Mabuchi
Takuma
T
Matsushita
Kohji
K
Yagita
Yoshiki
Y
Hori
Masatsugu
M
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002545:Brain Ischemia / Q000628:therapy*; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D004396:Coloring Agents / Q000494:pharmacology; D018076:DNA, Complementary / Q000378:metabolism; D004195:Disease Models, Animal; D004815:Epidermal Growth Factor / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006133:Growth Substances / Q000378:metabolism; D006493:Heparin / Q000737:chemistry; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D020244:Infarction, Middle Cerebral Artery / Q000628:therapy; D036341:Intercellular Signaling Peptides and Proteins; D007763:Lac Operon; D051379:Mice; D018613:Microscopy, Confocal; D009389:Neovascularization, Pathologic; D018919:Neovascularization, Physiologic; D009474:Neurons / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
D004396:Coloring Agents; D018076:DNA, Complementary; D006133:Growth Substances; C587883:HBEGF protein, human; C587884:Hbegf protein, mouse; C587885:Hbegf protein, rat; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D004815:Epidermal Growth Factor; D006493:Heparin; D001973:Bromodeoxyuridine