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（由申请人提供）：本 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检测）与完善的基因敲除和行为小鼠模型相结合，以获得破坏 2-花生四烯酰甘油 (2-AG) 生物合成的病理生理学后果的全局视图 肥胖和炎症的小鼠模型。内源性大麻素 (endocannabinoid) 2-AG 是一种脂质递质，可激活 G 蛋白偶联受体 CB1 和 CB2，这也是大麻中精神活性成分 9-四氢大麻酚的目标。 CB1拮抗剂作为抗肥胖和抗糖尿病药物的临床活性凸显了内源性大麻素系统在肥胖相关代谢紊乱中的重要性。此后，临床前模型的补充研究表明，许多有益效果是由于阻断外周部位的 CB1 受体而产生的。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中的中枢和/或外周贡献。体内 2-AG 生物合成受到两种序列相关酶二酰甘油脂肪酶-α 和 β（分别为 DAGL α 和 DAGL β）的差异调节，这提供了通过选择性失活 DAGL 亚型来解偶联中枢和外周内源性大麻素信号传导的实验（以及最终翻译）机会。该提案将检验以下假设：DAGL α 和 DAGL β 生物合成参与能量稳态的 2-AG，并通过调节解剖学不同位点的 CB1 依赖性信号传导在能量平衡中发挥互补功能。具体目标是 1) 测量 DAGL 破坏小鼠的体内能量稳态参数和体重组成 2) 测量 DAGL 破坏小鼠的脂肪组织巨噬细胞积累以及 3) 确定 DAGL 失活对代谢综合征发生和进展的生理影响。我们的初步研究表明，尽管食物摄入量增加，但 DAGL α 显示出瘦表型，这提供了 2-AG 生物合成中枢和/或外周破坏后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 信号通路的可行性。该候选人的目标是将之前化学蛋白质组学、合成化学、定量蛋白质组学和代谢组学方面的培训与小鼠代谢和炎症模型的新指导培训相结合，包括对控制能量稳态和体重组成的代谢参数进行非侵入性测量，以及使用过继细胞转移和流式细胞术对脂肪组织巨噬细胞积累进行体内跟踪。

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