Direct analysis of lipolysis-mediated signaling events

直接分析脂肪分解介导的信号事件

• 批准号: 10336569
• 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
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10336569
• 关键词: Direct; analysis; lipolysis; mediated; signaling

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.

项目摘要/摘要 肥胖已达到流行比例，并与代谢性疾病的更大患病率相关 糖尿病和心血管疾病。而肥胖引起糖尿病的确切机制不是 总体而言，越来越明显的证据表明，人体传感脂质的正常过程受到破坏。这 无法正确检测脂质会导致脂肪毒性，并导致关键胰岛素敏感的效果 组织。这是一个重要的科学目标，以及这一NIH独立奖的目标，就是 了解细胞感知脂质并维持脂质稳态的机制。培训 此应用程序的组成部分是基于候选人对成像代谢和背景的兴趣和背景 代谢信号/能量感应，同时提供独特的环境来训练职业发展 与团队科学进行研究有关的活动。研究培训部分将使用 独特的工具集将使候选人独立于脂肪分解的直接影响 跨膜蛋白激酶A（PKA）信号传导和图像脂肪酸代谢。候选人将获得 具有磷蛋白质组学和脂肪组学以及超分辨率成像的全球分析技术的经验。 利用这些工具和培训，候选人将确定1）直接通过脂解产生的信号，以及 2）细胞内脂质运输和脂解衍生信号的动力学。这个目标的核心是 假设脂解的信号直接产生以维持脂质稳态，并且高度 动态的。奖励的研究组成部分将由以下具体目的完成：目标1： 识别直接通过脂解产生的信号。已知脂解会产生信号，但 这一点是脂解的直接作用与跨膜-PKA信号无法区分。利用 激活ABHD5（脂肪酶共激活蛋白）的新型合成配体AIM 1将通过 以下子-AIMS：1A：识别依赖ABHD5的脂质介质。利用脂质方法， 候选人将确定调节下游代谢的ABHD5产生的生物活性脂质。 1b：到 识别依赖ABHD5的激酶激活途径。使用磷蛋白质组学方法，候选者 将确定ABHD5直接结果的磷酸化事件，激酶和途径 激活。目标2：确定脂肪酸及其代谢产物的运输动力学和脂质 调解人。这将通过使用新开发的一般编码的荧光传感器来实现 这允许监测脂肪酸和脂肪酰基-COAS的暂时和空间动力学。 拟议的K00/K99与NIH和NIDDK的任务保持一致，并将训练诺言 科学家了解调节脂质体内稳态的机制，并有可能是 在肥胖期间被破坏，这是一个重大的公共卫生优先事项。