Functional Characterization of Diacylglycerol Lipases in Mammalian Physiology

二酰甘油脂肪酶在哺乳动物生理学中的功能表征

• 批准号: 8701001
• 负责人: Ku-Lung Hsu
• 金额: $ 12.83万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-15 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8701001
• 关键词: 1,2-diacylglycerol; 2-arachidonylglycerol; Address; Adipose tissue; Adoptive Cell Transfers; Adverse effects; Anabolism; Animal Model; Animals; Antidiabetic Drugs; Behavioral; Biochemical; Biological Assay; Body Temperature; Body Weight; CNR1 gene; Calories; Carbon Dioxide; Chemicals; Chemotaxis; Chronic; Clinical; Development; Diet; Diglycerides; Disease; Eating; Eicosanoids; Endocannabinoids; Energy Metabolism; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Ethanolamines; Fatty acid glycerol esters; Flow Cytometry; G-Protein-Coupled Receptors; Genetic; Goals; Health; Heating; Homeostasis; Hydrolase; Inflammation; Inflammatory; Inflammatory Response; K-Series Research Career Programs; Knock-out; Knockout Mice; Label; Lead; Lipids; Marijuana; Measurement; Measures; Mediating; Mentors; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolic syndrome; Metabolism; Monitor; Monoacylglycerol Lipases; Mus; Neuraxis; Neurons; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Obesity associated disease; Organism; Oxygen; Pain; Pathway interactions; Peripheral; Peritoneal Macrophages; Pharmaceutical Preparations; Phase; Phenotype; Physiological; Physiology; Pre-Clinical Model; Process; Production; Protein Isoforms; Proteomics; Regulation; Serine Hydrolase; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Synthesis Chemistry; System; Testing; Tetrahydrocannabinol; Therapeutic Effect; Tissues; Training; Transgenic Mice; anandamide; balance testing; cannabinoid receptor; endogenous cannabinoid system; energy balance; feeding; in vivo; inhibitor/antagonist; innovation; insight; insulin sensitivity; lipoprotein lipase; macrophage; metabolomics; monocyte; mouse model; novel therapeutics; overexpression; prevent; respiratory; small molecule

DESCRIPTION (provided by applicant): The overall goal of this K99/R00 career development award proposal is to integrate innovate chemical probes, including in vivo-active small-molecule inhibitors and specialized chemoproteomic assays, with well- established genetic knockout and behavioral mouse models to obtain a global view of the pathophysiological consequence of disrupting 2-arachidonoylglycerol (2-AG) biosynthesis in mouse models of obesity and inflammation. The endogenous cannabinoid (endocannabinoid) 2-AG is a lipid transmitter that activates the G- protein-coupled receptors CB1 and CB2, which are also targets for the psychoactive ingredient in marijuana, 9-tetrahydrocannabinol. The importance of the endocannabinoid system in obesity-associated metabolic disorders is highlighted by the clinical activity of CB1 antagonists as anti-obesity and anti-diabetic drugs. Complementary studies in preclinical models have since shown that many of the beneficial effects are due to blocking CB1 receptors at peripheral sites. These studies highlight the need for a deeper understanding of the central and/or peripheral contributions of the endocannabinoid system in regulating energy homeostasis as well as obesity-related disease states. 2-AG biosynthesis in vivo is differentially regulated by two sequence- related enzymes, diacylglycerol lipase-alpha and beta (DAGL alpha and DAGL beta, respectively), providing an experimental (and, eventually translational) opportunity to uncouple central and peripheral endocannabinoid signaling through selective inactivation of DAGL isoforms. This proposal will test the hypothesis that DAGL alpha and DAGL beta biosynthesize 2-AG involved in energy homeostasis and perform complementary functions in energy balance through regulation of CB1-dependent signaling at anatomically-distinct sites. The specific aims are to 1) measure energy homeostasis parameters and body mass composition in vivo in DAGL-disrupted mice 2) measure adipose tissue macrophage accumulation in DAGL-disrupted mice and 3) Determine the physiological effects of inactivating DAGLs in the development and progression of metabolic syndrome. Our preliminary studies show that DAGL alpha display a lean phenotype despite increased food intake, providing evidence of altered energy balance upon central and/or peripheral disruption of 2-AG biosynthesis. Our preliminary studies also show the feasibility of using DAGL-selective inhibitors to uncouple central and peripheral 2-AG signaling pathways in vivo. The candidate aims to combine previous training in chemoproteomics, synthetic chemistry, and quantitative proteomics and metabolomics with new mentored training in mouse models of metabolism and inflammation including non-invasive measurements of metabolic parameters governing energy homeostasis and body mass composition as well as in vivo tracking of adipose tissue macrophage accumulation using adoptive cell transfer and flow cytometry.

描述(申请人提供)：这项K99/R00职业发展奖提案的总体目标是将创新的化学探针，包括体内活性小分子抑制剂和专门的化学蛋白质组分析，与成熟的基因敲除和行为小鼠模型相结合，以获得对2-花生四烯基甘油(2-AG)生物合成中断在肥胖和炎症小鼠模型中的病理生理后果的全球视角。内源性大麻素(内源性大麻素)2-AG是一种脂类递质，激活G蛋白偶联受体CB1和CB2，这两个受体也是大麻中具有精神活性的成分9-四氢大麻酚的靶标。CB1拮抗剂作为抗肥胖和抗糖尿病药物的临床活性突显了内源性大麻素系统在肥胖相关代谢紊乱中的重要性。临床前模型中的补充研究表明，许多有益的影响是由于阻断了周围部位的CB1受体。这些研究强调，需要更深入地了解内源性大麻素系统在调节能量稳态以及肥胖相关疾病状态方面的中枢和/或外周贡献。体内2-AG的生物合成受两种序列相关酶--二酰甘油脂肪酶-α和β(分别为DAGLα和DAGLβ)的差异调节，通过选择性地灭活DAGL亚型，提供了一个实验(并最终翻译)解偶联中枢和外周内源性大麻信号的机会。这一建议将检验DAGLα和DAGLβ生物合成2-AG参与能量动态平衡的假设，并通过调节不同解剖部位的CB1依赖信号来实现能量平衡的互补功能。其具体目的是：1)测量DAGL干扰小鼠体内的能量稳态参数和体质量组成；2)测量DAGL干扰小鼠脂肪组织中巨噬细胞的聚集；3)确定DAGL失活在代谢综合征发生发展中的生理作用。我们的初步研究表明，尽管食物摄入量增加，DAGLα仍表现为瘦肉型，这提供了2-AG生物合成中枢和/或外周中断时能量平衡改变的证据。我们的初步研究还表明，使用DAGL选择性抑制剂在体内解偶联中枢和外周2-AG信号通路是可行的。该候选人的目标是将先前在化学蛋白质组学、合成化学、定量蛋白质组学和代谢组学方面的培训与小鼠新陈代谢和炎症模型的新指导培训结合起来，包括对控制能量稳态和身体质量组成的代谢参数的非侵入性测量，以及使用过继细胞转移和流式细胞术在体内跟踪脂肪组织巨噬细胞的聚集。

项目成果

Highly Selective, Reversible Inhibitor Identified by Comparative Chemoproteomics Modulates Diacylglycerol Lipase Activity in Neurons.

• DOI: 10.1021/jacs.5b04883
• 发表时间: 2015-07-15
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Baggelaar MP;Chameau PJ;Kantae V;Hummel J;Hsu KL;Janssen F;van der Wel T;Soethoudt M;Deng H;den Dulk H;Allarà M;Florea BI;Di Marzo V;Wadman WJ;Kruse CG;Overkleeft HS;Hankemeier T;Werkman TR;Cravatt BF;van der Stelt M
• 通讯作者: van der Stelt M