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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要 K99/R00 职业发展奖提案的总体目标是整合创新化学探针、包括体内活性小分子抑制剂和专门的化学蛋白质组学测定，具有良好的建立基因敲除和行为小鼠模型，以获得病理生理学的全局视图破坏肥胖小鼠模型中 2-花生四烯酰甘油 (2-AG) 生物合成的后果内源性大麻素 (endocannabinoid) 2-AG 是一种脂质递质，可激活 G- 蛋白质偶联受体 CB1 和 CB2，也是大麻中精神活性成分的目标， 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突出了该疾病。此后，临床前模型的补充研究表明，许多有益效果归因于这些研究强调需要更深入地了解 CB1 受体。内源性大麻素系统在调节能量稳态中的中枢和/或外周贡献以及与肥胖相关的疾病状态。体内 2-AG 生物合成受两个序列的差异调节。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（以及最终的转化）解开中枢和外周内源性大麻素信号传导的机会通过选择性失活 DAGL 同工型该提案将检验 DAGLα 和 DAGLα 的假设。 DAGLβ生物合成参与能量稳态的2-AG并在能量中发挥补充功能通过在解剖学上不同的位点调节 CB1 依赖性信号传导来实现平衡，其具体目标是 1) 测量 DAGL 破坏小鼠的体内能量稳态参数和体重成分 2) 测量 DAGL 破坏小鼠的脂肪组织巨噬细胞积累，并 3) 确定 DAGL 失活对代谢综合征发生和进展的生理影响。初步研究表明，尽管食物摄入量增加，DAGLα仍显示出瘦表型，从而提供 2-AG 生物合成的中枢和/或外周破坏导致能量平衡改变的证据。初步研究还表明使用 DAGL 选择性抑制剂来解偶联中枢和体内外周2-AG信号通路。候选人的目标是结合之前在化学蛋白质组学、合成化学和定量方面的培训蛋白质组学和代谢组学以及代谢和炎症小鼠模型的新指导培训包括对控制能量稳态和体重的代谢参数进行非侵入性测量使用过继细胞转移对脂肪组织巨噬细胞积累的组成以及体内跟踪和流式细胞术。