| Project/Area Number |
18K08853
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 55050:Anesthesiology-related
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| Research Institution | Shiga University of Medical Science |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
守村 敏史 滋賀医科大学, 動物生命科学研究センター, 准教授 (20333338)
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| Project Period (FY) |
2018-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | GTP Cyclohydrolase 1 / tetrahyrobiopterin / antibody / scfv / intrabody / GTP cyclohydrolase 1 / BH4 / conformational antibody / amyloid beta / 3D imaging / immunotherapy / scFv / peripheral ChAT / acetylcholine / serotonin / Pain / pChAT / GCH1 |
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| Outline of Final Research Achievements |
GTP cyclohydrolase 1 (GCH1) is the first and rate-limiting enzyme in the tetrahydrobiopterin (BH4) synthesis pathway. BH4 is a cofactor required for the biosynthesis of monoaminergic and nitrergic neurotransmitters. GCH1 is a pain marker and BH4 level is linked to chronic pain, individuals with deficient GCH1 and low availability in BH4 experience low neuropathic pain. During this project, we produced specific monoclonal antibodies against GCH1, cloned their sequence, and produce single-chain antibodies (scFv). scFv were expressed in cell culture as intrabody, and their potential to block GCH1 activity was tentatively evaluated. Our observation suggested that scFv expression as intrabody is possible, but the aggregation of intrabody in cell jeopardize their action. Improvement of intrabody efficiency by reducing their aggregative propensity was also studied. Developed methods and techniques are shown to be useful to also study highly aggregative proteins such as amyloid-beta 42.
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| Academic Significance and Societal Importance of the Research Achievements |
研究課題名：ラットの脊髄後根神経節ニューロン内の痛みマーカーに対して作られたsingle domain intrabodyを用いた免疫療法の慢性疼痛に対する評価
「痛みは避けられないが、でも苦しみは自分次第」・・・村上春樹・・・痛みをどのように感じるか、そのメカニズムを探ることは新規治療法を開発する上で極めて重要である。その目的のために、我々は痛みのマーカーであるGCH1に対する新たな抗体、疼痛を感じるメカニズムを細胞内で遮断する抗体をintrabody（細胞内産生抗体）として作る。
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