Direct analysis of lipolysis-mediated signaling

项目概要/摘要 肥胖已经达到流行病的程度，并与代谢紊乱的更大流行有关， 糖尿病和心血管疾病。虽然肥胖导致糖尿病的确切机制并不清楚， 越来越多的证据表明，人体正常的脂质感知过程被破坏了。这 不能正确地检测脂质可导致脂毒性，并在关键的胰岛素敏感性细胞中引起有害影响。 组织中因此，一个重要的科学目标，也是NIH独立之路奖的目标，是 了解细胞感知脂质的机制，从而维持脂质体内平衡。培训 该应用程序的组成部分建立在候选人的兴趣和背景成像代谢， 代谢信号/能量传感，同时提供一个独特的环境，在职业发展培训 与团队科学研究方法有关的活动。研究培训部分将利用 一套独特的工具，将允许候选人探测脂解的直接影响， 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 在磷酸化蛋白质组学和脂质组学以及超分辨率成像的全球分析技术方面的经验。 利用这些工具和培训，候选人将确定1）脂解直接产生的信号， 2)细胞内脂质运输和脂解衍生信号的动力学。这一目标的核心是 由脂解作用直接产生的信号起维持脂质体内平衡的作用， 动态的该奖项的研究部分将通过以下具体目标来实现：目标1： 识别脂肪分解直接产生的信号。已知脂肪分解会产生信号，但最多 此时，脂解的直接影响与跨膜PKA信号无法区分。利用 新的合成配体，激活ABHD 5，脂肪酶共激活蛋白，目标1将通过以下方式实现： 1a：鉴定ABHD 5依赖性脂质介质。利用脂质组学方法， 候选人将鉴定由ABHD 5产生的调节下游代谢的生物活性脂质。1b：至 鉴定ABHD 5依赖性激酶活化途径。使用磷酸化蛋白质组学方法， 将决定磷酸化事件，激酶和途径，是ABHD的直接结果5 activation.目的2：研究脂肪酸及其代谢产物和脂质的转运动力学 调解员这将通过使用新开发的基因编码荧光传感器来实现 其允许监测脂肪酸和脂肪酰辅酶A的时间和空间动态。的 拟议的K 00/K99与NIH和NIDDK的使命非常一致，将培养一个有前途的 科学家了解调节脂质体内平衡的机制，以及潜在的途径， 在肥胖期间中断，这是一个重要的公共卫生优先事项。

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.