2002 Fiscal Year Final Research Report Summary
Basic research of the new gene therapy for cerebral infarction
| Project/Area Number |
13670669
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurology
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| Research Institution | Juntendo University |
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Principal Investigator |
URABE takao Juntendo University, Neurology, Assistant professor, 医学部, 講師 (60291663)
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| Co-Investigator(Kenkyū-buntansha) |
MIZUNO yoshikuni Juntendo University, Neurology, Professor, 医学部, 教授 (30049043)
MOCHIZUKI hideki Juntendo University, Neurology, Assistant professor, 医学部, 講師 (90230044)
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| Project Period (FY) |
2001 – 2002
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| Keywords | Cerebral infarction / Ischemic neuronal death / Gene therapy / Apoptosis / Oxidative stress / Adeno-associated virus vector / Bone marrow transplantation / Microglia |
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| Research Abstract |
Analysis of expressing apoptotic associated proteins in animal model of focal brain ischemia
To investigate the serial change and cellular localization of overexpressed Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), as well as whether or not overexpression of GAPDH is related to ischemia/reperfusion-induced neuronal apoptosis, we performed immunohistochemical study in a rat middle cerebral artery occlusion (MCAO) and reperfusion model. The current study distinctly demonstrated that nuclear translocation of overexpressed GAPDH occurred in both the ischemic core and the penumbra area soon after focal ischemia, and might be an early marker of ischemia/reperfusion-induced apoptotic neuronal death. Our findings suggest that GAPDH may play a critical role in the progression and expansion of ischemic neuronal damage.
To assess the distribution of microglia/macrophages in cerebral ischemia, we performed permanent focal ischemia using bone-marrow chimera mice known to express enhanced green fl
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uorescent protein (EGFP). This study directly showed the migration and distribution of bone marrow-derived monocytes/macrophages and the relationship between resident microglia and infiltrated hematogenous element in ischemic mouse brain.
Concentration and purification of Adeno-associated virus vector (AAV)
At present, we are promoting that Calbindin D28k cDNA is inserted into AAV.
Injection into the brain of animal model
We have already finished that Apaf 1 cDNA has been inserted into AAV, and adjusted the virus titer. Motchizuki H, et al. has been reported that the AAV-derived Apaf 1 dominant negative inhibitor prevents MPTP induced neuronal death as antiapoptotic gene therapy. At present, we are investigating that the AAV-derived Apaf 1 dominant negative inhibitor is injected into the gerbil hippocampus using a stereotaxic apparatus as antiapoptotic gene therapy for cerebral infarction.
Evaluation of oxidative stress as the marker of cell death
To assess the efficacy of gene therapy for ischemic neuronal death, we performed immunochemical study using antibodies of 4-hydoxy-2-nonenal (HNE)-modified protein and 8-hydroxy-2'-deoxyguanosine (8-OhdG) as a marker for oxidative stress. Less
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[Publications] Mochizuki H, Hayakawa H, Migita M, Shibata M, Tanaka R, Suzuki A, Shimo-Nakanishi Y, Urabe T, Yamada M, Tamayose K, Shimada T, Miura M, Yoshikuni M: "An AAV-derived Apaf-1 dominant negative inhibitor prevents MPTP toxicity as antiapoptotic gene therapy for Parkinson's disease"Proc Natl Acad Sci USA. 98. 10918-10923 (2001)
Description
「研究成果報告書概要(欧文)」より
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