Molecular and cell biological studies on the production, aggregation and clearance of Alzheimer amyloid β peptides.

关于阿尔茨海默病淀粉样β肽的产生、聚集和清除的分子和细胞生物学研究。

• 批准号: 17200025
• 负责人: IWATSYBO Takeshi
• 金额: $ 29.87万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17200025/
• 关键词: Alzheimer's disease; γ-Secretase; Amyloid; amyloid binding protein; Aβ; アルツハイマー病; βアミロイド

The aim of this research project is to elucidate the process of production, accumulation and clearance of amyloid β peptides, that are regarded as pathogenic proteins in Alzheimer's disease (AD), thereby creating a novel concept for the disease-modifying therapy for AD. Through the studies of γ-secretase, a membrane protease that produces Aβ peptide, we tried to clarify the mechanism of Aβ production and to block the most upstream cause of AD. Specifically we developed novel γ-secretase inhibitors harboring an enone-moiety and analyzed the mechanism of action of these compounds. Using Cys scan approach, we showed that the 6/7 transmembrane domains of presenilin 1, the latter being the catalytic subunit of γ-secretase, form a hydrophilic catalytic pore structure, We further analyzed the structure of γ-secretase and a related protease, SPP, using single particle analysis. We showed that CLAC, a novel amyloid plaque component protein, inhibits Aβ fibril formation in vitro, and is involved in the compaction of amyloid plaques in the brains of transgenic mice. We also showed that LRP1 is involved in the clearance of All peptides across the blood brain barrier, using an immortalized cell line of brain endothelial cells. We thus elucidated the key steps of Aβ deposition, i.e., γ-secretase, amyloid binding protein and Aβ clearance, using cutting edge technologies including proteomic analysis, RNAi and small molecule screen, providing clues to the development of novel therapeutic strategies for AD.

本研究旨在阐明阿尔茨海默病（Alzheimer's disease，AD）致病蛋白β淀粉样肽（amyloid β peptides）的产生、积累和清除过程，从而为AD的疾病修饰治疗创造新的概念。通过对产生Aβ肽的膜蛋白酶γ-分泌酶的研究，试图阐明Aβ产生的机制，阻断AD最上游的致病因素。具体而言，我们开发了含有烯酮部分的新型γ-分泌酶抑制剂，并分析了这些化合物的作用机制。用半胱氨酸扫描法，我们发现早老素1的6/7跨膜区，即γ-分泌酶的催化亚基，形成一个亲水性的催化孔结构。我们进一步用单颗粒分析法分析了γ-分泌酶及其相关蛋白酶SPP的结构。我们发现，CLAC，一种新的淀粉样斑块组分蛋白，在体外抑制Aβ原纤维的形成，并参与转基因小鼠脑中淀粉样斑块的压实。我们还表明，LRP 1参与清除所有肽跨越血脑屏障，使用永生化细胞系的脑内皮细胞。因此，我们阐明了Aβ沉积的关键步骤，即，γ-分泌酶、淀粉样蛋白结合蛋白和Aβ清除，利用包括蛋白质组学分析、RNAi和小分子筛选在内的尖端技术，为开发AD的新治疗策略提供线索。

项目成果

Molecular Pathology of Alzheimer's Disease

• DOI: 10.3109/09540269309037798
• 发表时间: 2013-11
• 作者: R. Castellani;George Perry

Structure of the catalytic pore of γ-secretase probed by the accessibility of substituted cysteines

• DOI: 10.1523/jneurosci.3614-06.2006
• 发表时间: 2006-11-15
• 期刊: JOURNAL OF NEUROSCIENCE
• 影响因子: 5.3
• 作者: Sato, Chihiro;Morohashi, Yuichi;Iwatsubo, Takeshi
• 通讯作者: Iwatsubo, Takeshi

Immunoreactivity of phage library-derived human single-chain antibodies to amyloid β conformers in vitro

噬菌体文库衍生的人单链抗体对淀粉样β构象异构体的体外免疫反应性

• 发表时间: 2007
• 期刊: J Biochem 143
• 作者: Yoshihara T;et. al.
• 通讯作者: et. al.

Concise and short synthesis of functionalized 5,6-Dihydropyridin-2-ones by means of palladium(0)-catalyzed cross-coupling of ketene aminal phosphates.

通过钯 (0) 催化乙烯酮缩醛磷酸酯的交叉偶联，简洁、简短地合成功能化 5,6-二氢吡啶-2-酮。

• 发表时间: 2006
• 期刊: Heterocycles 70
• 作者: Yoshihara T;et. al.;Hori Y;Ogura T;Morohashi Y;Tomita T;Takahashi Y;Garino C;Fuwa H;Ozaki T;Sato C;Fuwa H
• 通讯作者: Fuwa H

C-terminal fragment of Presenilin is the molecular target of a dipeptidic γ-secretase-specific Inhibitor DAPT

早老素的 C 端片段是二肽 γ-分泌酶特异性抑制剂 DAPT 的分子靶点

• 发表时间: 2006
• 期刊: J Biol Chem 281
• 作者: Morohashi Y;Kan T;Tominari Y;Fuwa H;Okamura Y;Watanabe N;Sato C;Natsugari H;Fukuyama T;Iwatsubo T^*;Tomita T^*
• 通讯作者: Tomita T^*