| Project/Area Number |
23300131
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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 |
Neurochemistry/Neuropharmacology
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| Research Institution | Kyoto University (2013)
Shiga University of Medical Science (2011-2012) |
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Principal Investigator |
URUSHITANI Makoto 京都大学, 医学(系)研究科(研究院), 准教授 (60332326)
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| Co-Investigator(Kenkyū-buntansha) |
KITAHARA Ryo 立命館大学, 薬学部, 准教授 (70512284)
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| Co-Investigator(Renkei-kenkyūsha) |
ITO Hidefumi 和歌山県立医科大学, 医学部, 教授 (20250061)
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| Project Period (FY) |
2011-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥20,020,000 (Direct Cost: ¥15,400,000、Indirect Cost: ¥4,620,000)
Fiscal Year 2013: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2012: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
Fiscal Year 2011: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000)
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| Keywords | 筋萎縮性側索硬化症 / TDP-43 / 高圧力NMR / 抗体 / 凝集体 / 細胞モデル / 抗体療法 / NMR解析 / 免疫組織化学 / in vitroモデル / モデルマウス |
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| Research Abstract |
Using structural analyses of RNA-recognition motifs (RRM) of TDP-43, a ALS-linked protein, we identified a aberrant assembly interface in RRM1. The cysteine in the this region induced TDP-43 aggregates, cytosolic mislocalization, the incompetent RNA splicing, and cytotoxicity, which functionally and structurally mimicks TDP-43 proteinopathy. We also succeeded in cloning cDNA from murine hybridoma (3B12A), generating a monoclonal antibody recognizing the misofolded forms of TDP-43, and constructed mammalian expression plasmid for the single chain Fv (scFv). Since the commercially available transgenic mice expressing ALS-linked mutant TDP-43 (A315T) turned out to fail to replicate ALS phenotype, we newly establish the embryonic brain transfection system, using in-utero electroporation. The brain slice expressing aggregate prone mutant TDP-43 showed ALS-like TDP-43 aggregates, which could be tested to estimate the intrabody therapy.
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