Direct analysis of lipolysis-mediated signaling events

直接分析脂肪分解介导的信号事件

基本信息

批准号：
10353527
负责人：
Emilio Mottillo
金额：
$ 24.9万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10353527
关键词：
Acyl Coenzyme A Address Adipocytes Adipose tissue Area Award Biochemical Bioinformatics Biological Bypass Cardiovascular Diseases Cells Chemicals Cyclic AMP-Dependent Protein Kinases Data Detection Diabetes Mellitus Environment Epidemic Event Fatty Acids Fluorescence Microscopy Generations Goals Health Homeostasis Hydrolase Hydrolysis Image Imaging Device Inflammation Insulin Integral Membrane Protein Knowledge Lead Ligands Lipase Lipids Lipolysis Mediating Mediator of activation protein Mentorship Metabolic Metabolic Diseases Metabolism Microscopy Mission Molecular Monitor National Institute of Diabetes and Digestive and Kidney Diseases Nonesterified Fatty Acids Nuclear Receptors Obesity Pathway Analysis Pathway interactions Phosphorylation Phosphotransferases Prevalence Principal Investigator Process Production Proteins Proteomics Regulation Research Research Training Resolution Role Science Scientist Signal Pathway Signal Transduction Techniques Time Tissues Training Triglycerides United States National Institutes of Health Work body sense career development diabetes risk experience fatty acid metabolism fluorescence imaging improved inhibitor/antagonist insight interest lipid mediator lipid metabolism lipid transport lipidomics novel perilipin phosphoproteomics programs public health priorities sensor tool trafficking

项目摘要

Project Summary/Abstract Obesity has reached epidemic proportions and is tied to the greater prevalence of metabolic disorders such as diabetes and cardiovascular disease. While the precise mechanisms by which obesity causes diabetes are not entirely clear, mounting evidence suggests that the body’s normal process of sensing lipids is disrupted. This inability to properly detect lipids can lead to lipotoxicity and cause detrimental effects in key insulin sensitive tissues. Thus, an important scientific goal, and that of this NIH Pathway to Independence Award, is to understand the mechanisms by which cells sense lipids and thereby maintain lipid homeostasis. The training component of this application builds upon the candidate’s interest and background in imaging metabolism and metabolic signaling/energy sensing, while providing a unique environment to train in career development activities related to a team science approach of doing research. The research training component will utilize a unique set of tools that will allow the candidate to probe the direct effects of lipolysis independent of transmembrane-protein kinase A (PKA) signaling and image fatty acid metabolism. The candidate will gain experience in global analysis techniques of phosphoproteomics and lipidomics, and super-resolution imaging. Utilizing these tools and training, the candidate will determine 1) the signals directly generated by lipolysis, and 2) the dynamics of lipid trafficking and lipolysis-derived signals within a cell. Central to this aim is the hypothesis that signals directly produced by lipolysis function to maintain lipid homeostasis and are highly dynamic. The research component of the award will be accomplished by the following specific aims: Aim 1: Identification of signals that are generated directly by lipolysis. Lipolysis is known to produce signals, but up to this point the direct effects of lipolysis were not distinguishable from transmembrane-PKA signals. Utilizing novel synthetic ligands that activate ABHD5, a lipase co-activator protein, Aim 1 will be accomplished by the following sub-aims: 1a: To identify ABHD5-dependent lipid mediators. Utilizing a lipidomic approach, the candidate will identify the bioactive lipids produced by ABHD5 that regulate downstream metabolism. 1b: To identify ABHD5-dependent kinase activation pathways. Using a phosphoproteomic approach, the candidate will determine the phosphorylation events, kinases and pathways that are a direct consequence of ABHD5 activation. Aim 2: To determine the trafficking dynamics of fatty acids and their metabolites and lipid mediators. This will be accomplished by the use of newly developed genetically encoded fluorescent sensors that allow the monitoring of the temporal and spatial dynamics of fatty acids and fatty acyl-CoAs. The proposed K00/K99 is well aligned with the mission of the NIH and the NIDDK and will train a promising scientist to understand the mechanisms that regulate lipid homeostasis and potentially the pathways that are disrupted during obesity, a significant public health priority.

项目概要/摘要肥胖已达到流行病的程度，并且与代谢性疾病的流行有关，例如虽然肥胖导致糖尿病的确切机制尚不清楚。完全清楚，越来越多的证据表明，身体感知脂质的正常过程被破坏了。无法正确检测脂质可能会导致脂毒性，并对关键的胰岛素敏感人群造成痛苦影响因此，一个重要的科学目标，也是 NIH 独立之路奖的目标，是了解细胞感知脂质并从而维持脂质稳态的机制。该应用程序的组成部分建立在候选人对成像代谢的兴趣和背景之上代谢信号/能量传感，同时提供独特的职业发展培训环境与进行研究的团队科学方法相关的活动。研究培训部分将利用一套独特的工具，使候选人能够探索脂肪分解的直接影响，独立于跨膜蛋白激酶 A (PKA) 信号传导和脂肪酸代谢图像。在磷酸蛋白质组学和脂质组学以及超分辨率成像的整体分析技术方面拥有丰富的经验。利用这些工具和培训，考生将确定 1) 脂肪分解直接产生的信号，以及 2）细胞内脂质运输和脂解衍生信号的动态是这一目标的核心。假设脂肪分解产生的信号直接发挥作用以维持脂质稳态，并且高度该奖项的研究部分将通过以下具体目标来实现：目标 1：脂肪分解直接产生的信号的识别已知会产生信号，但最多可达。此时，脂肪分解的直接影响与跨膜-PKA 信号无法区分。激活 ABHD5（一种脂肪酶共激活蛋白）的新型合成配体，目标 1 将通过以下子目标： 1a：利用脂质组学方法鉴定 ABHD5 依赖性脂质介质。候选人将鉴定 ABHD5 产生的调节下游代谢的生物活性脂质 1b: To。使用磷酸蛋白质组学方法识别 ABHD5 依赖性激酶激活途径。将确定 ABHD5 直接后果的磷酸化事件、激酶和途径目标 2：确定脂肪酸及其代谢物和脂质的运输动态。这将通过使用新开发的基因编码荧光传感器来实现。允许监测脂肪酸和脂肪酰辅酶A的时间和空间动态。拟议的 K00/K99 与 NIH 和 NIDDK 的使命高度契合，并将培养有前途的人才科学家了解调节脂质稳态的机制以及潜在的途径肥胖期间受到干扰，这是一个重要的公共卫生优先事项。