Lipid Mediators and Lipid Metaborome

脂质介质和脂质代谢组

• 批准号: 15002004
• 负责人: SHIMIZU Takao
• 金额: $ 443.46万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Specially Promoted Research
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15002004/
• 关键词: Lipid mediator; Acyltransferase; Platelet-activating factor; Phospholipase A2; G protein-coupled receptor; Knockout mice (or Gene-deficient mice); Metabolome; Pulmonary surfactant lipids; アシル転位酵素; ロイトコイエン; 質量分析計; ロイトコリエン; Gタンパク共役型受容; ロイコトリエン

1 Functional analysis of lysophospholipid acyltransferase ; We identified several lysophospholipid acyltransferases involving in the remodeling pathway of membrane lipids biosynthesis by in silico database analyses. Based on these information, we succeeded in molecular cloning of lyso-PAF acetyltransferase and LPCAT1. Lyso-PAF acetyltransferase is a critical enzyme in stimulus dependent formation of PAR LPCAT1 is an acetyltransferase involving in synthesis of phosphatidylcholine in pulmonary surfactant and play a pivotal role in respiratory physiology. We are generating gene-deficient mice of these enzymes to clarify the physiological importance. We further analyze other enzymes in this family.2 Functional analysis of phospholipase A_2 (PLA_2) ; Among many mammalian PLA_2s, cytosolic PLA_2 α(cPLA_2α) plays a critical role in various physiological and pathophysiological conditions through generating lipid mediators. We newly discovered cPLA_2 family enzymes, cPLA_2δ, cPLA_2ε, and cPLA_2 … More ζ . We are analyzing the physiological function of these enzymes.3 Identification of novel lipid mediator receptors (G-protein coupled receptors : GPCRs) ; To search for novel lipid mediator receptors among approx. 150 orphan GPCRs, we succeeded in selecting approx. 30 candidate GPCRs. In these GPCRs, we have newly identified TDAG8 and G2A as proton-sensing receptors and LPA4 as a lysophosphatidic acid receptor. We are continuously performing the ligand screening for other orphan GPCRs.4 Analysisi of GPCR-deficient mice ; To clarify the physiological significance of lipid mediator GPCRs, we have generated genedeficient mice for BLT1, BLT2, CysLT2 and LPA4. In the analysis of BLT1-deficient mice, we obtained evidence that BLT1 is necessary for production of Th2 cytokines and the development of airway hyperresponsiveness in an OVA-induced bronchial asthma model.5 High throughput quantitation of lipid mediators by LC-MS ; By the combination of lipid chromatography and tandem mass spectrometry (MS/MS), we developed a highly specific and reasonably sensitive assay procedure to simultaneously quantitate > 14 different mediators in 10 min single run. Less

1 溶血磷脂酰基转移酶的功能分析；我们通过计算机数据库分析鉴定了几种参与膜脂生物合成重塑途径的溶血磷脂酰基转移酶。基于这些信息，我们成功地分子克隆了lyso-PAF乙酰转移酶和LPCAT1。 Lyso-PAF 乙酰转移酶是 PAR 刺激依赖性形成中的关键酶。 LPCAT1 是参与肺表面活性物质中磷脂酰胆碱合成的乙酰转移酶，在呼吸生理学中发挥着关键作用。我们正在培育这些酶的基因缺陷小鼠，以阐明其生理重要性。我们进一步分析了该家族中的其他酶。2磷脂酶A_2（PLA_2）的功能分析；在许多哺乳动物PLA_2中，胞质PLA_2α(cPLA_2α)通过产生脂质介质在各种生理和病理生理条件中发挥关键作用。我们新发现了 cPLA_2 家族酶，cPLA_2δ、cPLA_2ε 和 cPLA_2 … 更多 ζ 。我们正在分析这些酶的生理功能。3新型脂质介质受体（G蛋白偶联受体：GPCR）的鉴定；在大约中寻找新的脂质介质受体。 150个孤儿GPCR，我们成功地选择了大约。 30 个候选 GPCR。在这些 GPCR 中，我们新鉴定出 TDAG8 和 G2A 作为质子敏感受体，LPA4 作为溶血磷脂酸受体。我们正在不断对其他孤儿GPCR进行配体筛选。4 GPCR缺陷小鼠的分析；为了阐明脂质介质 GPCR 的生理意义，我们培育了 BLT1、BLT2、CysLT2 和 LPA4 基因缺陷小鼠。在对 BLT1 缺陷小鼠的分析中，我们获得了证据，表明 BLT1 对于 OVA 诱导的支气管哮喘模型中 Th2 细胞因子的产生和气道高反应性的发展是必需的。5 通过 LC-MS 对脂质介质进行高通量定量；通过结合脂质色谱和串联质谱 (MS/MS)，我们开发了一种高度特异性且相当灵敏的测定程序，可在 10 分钟单次运行中同时定量 > 14 种不同的介质。较少的

项目成果

Cytosolic phospholipase A2 alpha-deficient mice are resistant to experimental autoimmune encephalomyelitis.

缺乏实验性自身免疫性脑脊髓炎的胞质磷脂酶A2α缺陷型小鼠具有抗性。

• DOI: 10.1084/jem.20050665
• 发表时间: 2005-09-19
• 期刊: JOURNAL OF EXPERIMENTAL MEDICINE
• 影响因子: 15.3
• 作者: Marusic, S;Leach, MW;Pelker, JW;Azoitei, ML;Uozumi, N;Cui, JQ;Shen, MWH;DeClercq, CM;Miyashiro, JS;Carito, BA;Thakker, P;Simmons, DL;Leonard, JP;Shimizu, T;Clark, JD
• 通讯作者: Clark, JD

Identification of the lntracellular region of the Leukotriene B4 receptor type-1 tnat specifically involves in Gi-activation.

1 型白三烯 B4 受体 tnat 细胞内区域的鉴定特别涉及 Gi 激活。

• 发表时间: 2007
• 期刊: J. Biol. Chem. 282
• 作者: Kuniyeda;K.;Okuno;T.;Terawaki;K.;Miyano;M.;Yokomizo;T.;Shimizu;T.
• 通讯作者: T.

Cytosolic Phospholipase A2 ・ deficient Mice Are Resistant to Collagen-induced Arthritis

胞质磷脂酶 A2·缺陷小鼠对胶原蛋白诱导的关节炎有抵抗力

• 发表时间: 2003
• 期刊: J. Exp. Med. 197
• 作者: Hegen;M.;Sun L.;Uozumi N.;Kume K.;Goad;E. M.;Nickerson-Nutter L. C.;Shimizu T.;Clark D. J.
• 通讯作者: Clark D. J.

Lipid mediators ; fundamental regulators of immune responses and inflammatory reactions.

脂质介质；

• 发表时间: 2006
• 作者: Kabeya;Y;M. Shibasaki;清水孝雄
• 通讯作者: 清水孝雄

Characterization and Biological Roles of Leukotrie B4 Receotors.

Leukotrie B4 受体的特征和生物学作用。

• 发表时间: 2003
• 作者: Y.Ogasaka;K.Yamashita;H.Kunieda et al.;Hideo Shindou;Kamada Y;Takao Shimizu
• 通讯作者: Takao Shimizu