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: 9109601
• 负责人: Ku-Lung Hsu
• 金额: $ 24.65万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-15 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9109601
• 关键词: 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; CNR2 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; Heating; High Fat Diet; 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; Transmembrane Domain; abstracting; anandamide; arachidonate; balance testing; chemoproteomics; diabetic; endogenous cannabinoid system; energy balance; feeding; in vivo; inhibitor/antagonist; innovation; insight; insulin sensitivity; lipoprotein lipase; macrophage; metabolic phenotype; metabolomics; monocyte; mouse model; novel therapeutics; overexpression; prevent; respiratory; small molecule inhibitor

Project Summary/Abstract 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-α and β (DAGLα and DAGLβ, 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 DAGLα and DAGLβ 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 DAGLα 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.

项目摘要/摘要 该K99/R00职业发展奖提案的总体目标是整合创新的化学问题， 包括体内活性的小分子抑制剂和专门的化学蛋白质组学测定，具有良好的 已建立的遗传基因敲除和行为小鼠模型，以获得病理生理的全局视图 在肥胖和 炎。内源性大麻素（内源性大麻素）2-AG是一种脂质发射器，可激活G- 蛋白质偶联受体CB1和CB2，它们也是大麻中精神活性成分的靶标， Δ9-四氢大麻酚。内源性大麻素系统在肥胖相关代谢中的重要性 CB1拮抗剂作为抗肥胖和抗糖尿病药物的临床活性强调了疾病。 此后，临床前模型中的互补研究表明，许多有益作用是由于 阻断外围部位的CB1受体。这些研究强调了需要更深入地了解 内源性大麻素系统在调节能量稳态中的中央和/或外周贡献为 以及与肥胖相关的疾病状态。 2-AG生物合成体内由两个序列差异调节 相关酶，二酰基甘油脂肪酶-α和β（分别为daglα和daglβ），提供了实验性 （并且最终翻译）有机会揭示中央和周围内源性大麻素信号传导 通过选择性灭活DAGL同工型。该提案将检验以下假设 DAGLβ生物合成2-Ag参与能量稳态并在能量中执行完整功能 通过调节在解剖赋予位点的CB1依赖性信号传导的平衡。具体目的是 1）测量dagl disted小鼠体内体内的能量稳态参数和体重组成2） 测量脂肪组织巨噬细胞在dagl disted小鼠中的积累，3）确定 灭活dagls在代谢综合征发展和进展中的生理影响。我们的 初步研究表明，daglα显示出瘦表型的欲望增加食物摄入量，提供 2 AG生物合成的中央和/或外围破坏时能量平衡改变的证据。我们的 初步研究还表明，使用DAGL选择性抑制剂使中心和 体内外围2-AG信号通路。 候选人旨在结合化学蛋白质组学，合成化学和定量的先前培训 蛋白质组学和代谢组学具有新的新陈代谢和注射鼠标模型中的新培训 包括对能量稳态和体重的代谢参数的非侵入性测量 使用自适应细胞转移的脂肪组织巨噬细胞积累的组成以及体内跟踪 和流式细胞仪。