| Project/Area Number |
14370553
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Ophthalmology
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| Research Institution | Tohoku University |
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Principal Investigator |
ABE Toshiaki Tohoku University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (90191858)
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| Co-Investigator(Kenkyū-buntansha) |
WADA Yuko Tohoku University Hospital, Lecture, 病院・講師 (70302130)
TOMITA Hiroshi Tohoku University, Graduate School of Medicine, Research associate, 大学院・医学系研究科, 助手 (40302088)
SATO Hajime Tohoku University Hospital, Research associate, 病院・助手 (10312571)
YAMATO Masayuki Tokyo Women's Medical University, Lecture, 医学部, 講師 (40267117)
OKANO Mitsuo Tokyo Women's Medical University, Professor, 医学部, 教授 (00130237)
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| Project Period (FY) |
2002 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2005: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2004: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2003: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2002: ¥7,400,000 (Direct Cost: ¥7,400,000)
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| Keywords | retinal pigment epithelium / iris pigment epithelium / brain derived-neurotrophic factor / adeno-associated virus / cell transplantation / TrkB / アデノアソシエイトウイルス / 免疫染色 / in situ hybridization / 網膜変性 / 光障害 / ウイルス至適濃度 / 神経保護因子 / バリアー / ウイルスベクター / 細胞増殖因子遺伝子 / 加齢黄斑変性 / 網膜色素変性 / 主要組織適合抗原 |
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| Research Abstract |
We report a subretinal iris pigment epithelial cell (IPE) transplantation with safely. When these cells were genetically engineered that expressing neurotrophic factors, the transplantation rescued the photoreceptor cells from phototoxicity. We used adeno-associate virus (AAV2) for delivering neurotrophic factors, such as brain-derived neurotrophic factor (BDNF) to the transplanted cells. If we used more than 1x107 capsides/ml AAV-BDNF for transfection, statistically significant photoreceptor protection was observed in the AAV-BDNF-IPE transplantation from phototoxicity. However, the rescue effect was not dose-dependent. Then, we examined the BDNF receptor TrkB expression, because the receptor may control the effect of BDNF. So far many isoforms of TrkB have reported and we examined two of the major isoforms of TrkB ; TrkB-FL which has tyrosine kinase activity in the cell and TrkB-T1 which has not. These isoforms showed not always completely same expression by immunohistochemistry. TrkB-FL was expressed mainly on nerve fiber layer, ganglion cells layer, and inner nuclear layer (INL), conversely TrkB-T1 was on INL and RPE. Both isoforms were not expressed in ONL and photoreceptor layer. From the results of double immunostaining with S100β, These isforms were expressed on Muller cells. Further these isoforms were expressed in the retina not only spatially but also temporally different way, although the expression was enhanced by the AAV-BDNF-IPE transplantation. When we performed in situ hybridization, same expression pattern were also confirmed. siRNA experiments showed significant less photoreceptor protection when we injected siRNA of TrkB-T1. When we examined differential gene expression by DNA microarray of Muller cell line, rMC-1 between BDNF stimulated and non-stimulated, neurotrophic factors that were so far reported were not many expressed. Another mechanism such as scavenging the ions in the surrounding circumstances may be important.
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