A novel functional genomics approach and ligand hunting for orphan seven transmembrane receptors.

一种新颖的功能基因组学方法和寻找孤儿七跨膜受体的配体。

• 批准号: 15590095
• 负责人: HIRASAWA Akira
• 金额: $ 2.37万
• 依托单位: Kyoto University (2004)National Research Institute for Child Health and Development (2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590095/
• 关键词: orphan receptor; ligand hunting; GPR120; GLP-1; enteroendocrine cells; obesity; diabetes; GFP; データベース; 細胞内局在

This research aims to establish a novel functional genomics approach and ligand hunting method for orphan receptor by combining technology of subcellular localization analysis for receptor, signal transduction analysis, and expression profile analysis.Following three experiments were done.1.Ligand hunting for orphan receptor GPR120To identify endogenous ligands for GPR120, we performed receptor internalization assay established in this research project and we found that long-chain free fatty acid evoke specific internalization of GPR120. In agreement with internalization assay, long-chain fatty acids were found to evoke specific rise in [Ca^<2+>]_i and ERK activation.2.Expression profile analysis for GPR120To clarify the function of the orphan receptor GPR120 I tried the gene expression profile analysis. Using microarray analysis, I found that GPR120 express abundantly in intestines and enteroendocrine cell line STC-1.3.Functional analysis for GPR120 in GPR120 natively expressing cell line STC-1In STC-1 cells, free fatty acids stimulation evokes GLP-1 secretion, rise in [Ca^<2+>]_i and ERK activation. Transfection of RNA_i specific for GPR120, inhibited GLP-1 secretion and [Ca^<2+>]_i rise. These results indicate that fatty ac_id induced Ca^<2+> increase and GLP-1 secretion in STC-1 cell is mediated through GPR120.The discovery of ligand and physiological role for GPR120 prove the effectiveness of the technique established in this research. The purpose of this research was achieved.

本研究旨在通过结合受体亚细胞定位分析、信号转导分析和表达谱分析等技术，建立一种新的孤儿受体功能基因组学方法和配体搜寻方法，主要完成了以下三个实验：1.孤儿受体GPR 120的配体搜寻为了鉴定GPR 120的内源性配体，我们进行了本研究项目中建立的受体内化测定，我们发现长链游离脂肪酸引起GPR 120的特异性内化。长链脂肪酸可引起[Ca^<2+>]_i和ERK激活的特异性升高。2. GPR 120的表达谱分析为了阐明孤儿受体GPR 120的功能，我尝试了基因表达谱分析。3.天然表达GPR 120的STC-1细胞株中GPR 120的功能分析在STC-1细胞中，游离脂肪酸刺激可引起GLP-1分泌，[Ca^2+] i升高，ERK激活。转染GPR 120特异性RNA_i可抑制GLP-1分泌和[Ca^<2+>]_i升高。这些结果表明，脂肪酸诱导STC-1细胞内Ca^2+升高和GLP-1分泌是通过GPR 120介导的。GPR 120配体的发现及其生理作用证明了本研究建立的技术的有效性。达到了本研究的目的。

项目成果

Fukuhara Y, Hirasawa A, Tsujimoto G, et al.: "Gene expression profiles in the regenerating rata liver after partial hepatectomy."J Hepatology. 38. 784-792 (2003)

Fukuhara Y、Hirasawa A、Tsujimoto G 等人：“部分肝切除术后再生肝脏中的基因表达谱。”J Hepatology。

Yamada M, Hirasawa A, Shiojima S, Tsujimoto G: "Granzyme A mediates glucocorticoid-induced apoptosis in leukemia cells."FASEB Journal.. 17(12). 1712-1714 (2003)

Yamada M、Hirasawa A、Shiojima S、Tsujimoto G：“粒酶 A 介导白血病细胞中糖皮质激素诱导的细胞凋亡。”FASEB Journal.. 17(12)。

Katsuma S, Shiojima S, Hirasawa A, Tanoue A, Yano J, Tsujimoto G, et al.: "Transcriptional profiling of gene expression patterns during sphingosine 1-phosphate-induced mesangial cell proliferation"Biochem Biophys Res Commun.. 300. 577-584 (2003)

Katsuma S、Shiojima S、Hirasawa A、Tanoue A、Yano J、Tsujimoto G 等人：“1-磷酸鞘氨醇诱导的系膜细胞增殖过程中基因表达模式的转录分析”Biochem Biophys Res Commun.. 300. 577-

Richardson J, Chatwin H, Hirasawa A, Tsujimoto G, Evans PD: "Agonist-specific coupling of a cloned human alpha(1A)-adrenoceptor to different second messenger pathways."Naunyn Schmiedebergs Arch Pharmacol.. 367. 333-341 (2003)

Richardson J、Chatwin H、Hirasawa A、Tsujimoto G、Evans PD：“克隆人 α(1A)-肾上腺素受体与不同第二信使途径的激动剂特异性偶联。”Naunyn Schmiedebergs Arch Pharmacol.. 367. 333-341 (2003)

Gene expression profile in experimental mesangial proliferative glomerulonephritis.

• DOI: 10.1254/jphs.rc0040012
• 发表时间: 2004
• 期刊: Journal of pharmacological sciences
• 影响因子: 3.5
• 作者: Y. Izumi;Yasuhiro Izumiya;M. Shiota;T. Yukimura;S. Shiojima;M. Yamada;T. Kadowaki;S. Katsuma;A. Hirasawa;G. Tsujimoto;H. Iwao