Identification of ligand and function of novel group of free fatty acid receptors by using genome information.

利用基因组信息鉴定新型游离脂肪酸受体的配体和功能。

• 批准号: 17209003
• 负责人: TSUJIMOTO Gozoh
• 金额: $ 33.11万
• 依托单位: Kyoto University
• 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 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17209003/
• 关键词: Orphan G protein-coupled; GPR120; Enteroendocrine cell; Free fatty acid; Intracellular calcium; GLP-1 (glucagon-like peptide 1); GPR 40 family (GPR40,41,43); G蛋白質共役型受容体; GPR40; 抗体; カルシウムシグナル; ERK; ノックアウトマウス; 遊離脂肪酸受容体; リガンド; GLP-1

The result achieved for this research is summarized as following. The following research items planned at first were performed.(1)Cloning of free fatty acid receptor family(1)We cloned orphan G protein-coupled receptor GPR120 from genome database, and successfully de-orphanized ; hence, the natural ligand of GPR120 was free fatty acid. In this research, we performed cloning of all the other free fatty acid receptors (GPR40,GPR41,GPR43) by fully taking advantage of the genome database (human, mouse, and rat).(2)Development of tools for in vivo/in vitro functional analysisWe succeeded in making specific antibodies agains to each free fatty acid receptor. Moreover, detailed information of tissue distribution of each receptor was collected. In addition, various compounds were screened by using a HTS signal transduction analysis platform, and found natural products specific to each receptor was found. Furthermore, we cloned mouse genomes of each receptor, and clarified the structure, and tr … More ied to generate genetically engineered animals (especially, knockout mouse), and eventually for part of the receptor family, we succeeded in generating knockout mouse.(3)GPR120 function analysisThe in vitro function was analyzed by using native cell line STC-1 cell which are expressing GPR120, and we clarified that GPR120 plays a key role in regulating the secretion of incretin hormone GLP-1, which is important factor for insulin secretion. Moreover, we clarified that the GPR120-mediated secretion of GLP-1 is via the calcium and ERK signaling mechanism. Also, this GPR120 receptor-mediated signal transduction suppresses the apoptosis in this enteroendocrine cells.SummaryWe successfully prepared a various tools (e.g., specific antibodies, chemical probes, and knockout mice, etc.) to further analyze the physiological roles of free fatty acid receptor family. Based on this preparations, the physiological and pathophysiological role of these free fatty acid receptor family will be further clarified, and novel drug discovery can be expected in the future. Less

本研究取得的成果总结如下。首先进行了以下研究项目：(1)游离脂肪酸受体家族的克隆(1)从基因组数据库中克隆了孤儿G蛋白偶联受体GPR120，并成功去孤儿化；因此，GPR120的天然配体是游离脂肪酸。在本研究中，我们充分利用基因组数据库（人、小鼠和大鼠）对所有其他游离脂肪酸受体（GPR40、GPR41、GPR43）进行了克隆。（2）体内/体外功能分析工具的开发我们成功地制备了针对每种游离脂肪酸受体的特异性抗体。此外，还收集了每种受体的组织分布的详细信息。此外，利用HTS信号转导分析平台筛选了多种化合物，发现了针对每种受体特异的天然产物。此外，我们还克隆了小鼠各受体的基因组，并阐明了其结构，并尝试生成基因工程动物（特别是基因敲除小鼠），最终针对部分受体家族，成功生成了基因敲除小鼠。 (3)GPR120功能分析利用表达GPR120的天然细胞系STC-1细胞进行体外功能分析，阐明GPR120在基因工程中发挥着关键作用。 调节肠促胰岛素激素GLP-1的分泌，GLP-1是胰岛素分泌的重要因素。此外，我们还阐明 GPR120 介导的 GLP-1 分泌是通过钙和 ERK 信号传导机制实现的。此外，这种GPR120受体介导的信号转导抑制了这种肠内分泌细胞的凋亡。总结我们成功地制备了各种工具（例如特异性抗体、化学探针和基因敲除小鼠等）来进一步分析游离脂肪酸受体家族的生理作用。基于该制剂，这些游离脂肪酸受体家族的生理和病理生理作用将得到进一步阐明，未来新的药物发现可期。较少的

项目成果

From the cover : V1a Vasopressin receptors maintain normal blood pressure by regulating circulating blood volume and baroreflex sensitivity.

封面图片：V1a 加压素受体通过调节循环血量和压力反射敏感性来维持正常血压。

• 发表时间: 2006
• 期刊: Proc. Natl. Acad. Sci. U S A. Vol.103
• 作者: Koshimizu TA;Nasa Y;Tanoue A;Oikawa R;Kawahara Y;Kiyono Y;Adachi T;Tanaka T;Kuwaki T;Mori T;Takeo S;Okamura H;Tsujimoto G
• 通讯作者: Tsujimoto G

Function of the lower urinary tract in mice lacking alpha 1 d-adrenoceptor.

缺乏α1D-肾上腺素受体的小鼠下尿路的功能。

• 发表时间: 2005
• 期刊: J. Urol. Vol.174
• 作者: Chen Q;Takahashi S;Zhong S;Hosoda C;Zheng HY;Ogushi T;Fujimura T;Ohta N;Tanoue A;Tsujimoto Kitamura T.
• 通讯作者: Tsujimoto Kitamura T.

A probabilistic model for mining implicit 'chemical compound-gene' relations from literature

• DOI: 10.1093/bioinformatics/bti1141
• 发表时间: 2005-01
• 期刊: Bioinformatics
• 影响因子: 5.8
• 作者: Shanfeng Zhu;Y. Okuno;G. Tsujimoto;Hiroshi Mamitsuka

Correlation between vasoconstrictor roles and mRNA expression of alpha(1)-adrenoceptor subtypes in blood vessels of genetically engineered mice.

基因工程小鼠血管中血管收缩作用与 α(1)-肾上腺素受体亚型 mRNA 表达之间的相关性。

• 发表时间: 2005
• 期刊: Br.J.Pharmacol. 146
• 作者: M.Isaji et al.;H.Wang et al.;N.Yamada et al.;H.Ueno et al.;H.Tanahashi et al.;Okuno Y;Adachi T;Ishida A;Hosoda C
• 通讯作者: Hosoda C

スタンダード薬学シリーズ1「日本薬学会編 ヒューマニズム・薬学入門」

标准药学系列1《人文主义/药学导论》日本药学会编

• 发表时间: 2005
• 作者: Okuno Y;Yang J;Taneishi K;Yabuuchi H;Tsujimoto G GLIDA;Ishida A;Hosoda C;Zhu S;Lazaro-Suarez ML;Kagaya S;Zacharia J;Sugimoto Y;Nagata N;Egashira N;Kagaya S;Chen Q;Erami C;Katsuma S;Adachi T;Katsuma S;Yamada M;Yamauchi J;Hosoda C;Adachi T;Katsuma S;Deighan C;Hirasawa A;Ami Y;Koshimizu T;辻本 豪三
• 通讯作者: 辻本 豪三