Identifying the receptors of environmentally sensitive epoxy-eicosanoids with AMS

使用 AMS 识别环境敏感环氧类二十烷酸的受体

• 批准号: 9388619
• 负责人: Kin Sing Stephen Lee
• 金额: $ 24.9万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-15 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9388619
• 关键词: Absence of pain sensation; Acids; Affect; Area; Aryl Hydrocarbon Receptor; Award; Basic Science; Binding; Biological; Biological Assay; Biological Process; Biological Sciences; Biology; Biomedical Research; Blood Pressure; C14 isotope; California; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular Physiology; Cardiovascular system; Cell Surface Receptors; Cells; Cellular biology; Chemicals; Communities; Competitive Binding; Complement; Core Facility; Coupled; Cytochrome P450; Data; Diet; Ecology; Educational process of instructing; Educational workshop; Eicosanoids; Environmental Exposure; Enzymes; Epoxide hydrolase; Epoxy Compounds; Exposure to; Faculty; Fatty Acids; Fibrosis; G-Protein-Coupled Receptors; Genomics; Goals; Grant; Health; Human; Hypertension; In Vitro; Individual; Inflammation; Institution; International; Knowledge; Label; Lead; Leadership; Learning; Ligands; Lipids; Liquid substance; Malignant Neoplasms; Mediator of activation protein; Mentors; Metabolism; Methodology; Methods; Molecular; Molecular Biology; Neonatal; Omega-3 Fatty Acids; Omega-6 Fatty Acids; Pain; Pathway interactions; Peptides; Perception; Peroxisome Proliferators; Pesticides; Phase; Physiological; Play; Poison; Protein Chemistry; Proteomics; Public Health; Publications; Radiolabeled; Records; Research; Research Personnel; Rhodopsin; Role; Running; Science; Scientist; Scintillation Counting; Signal Pathway; Signal Transduction; Signaling Molecule; Smooth Muscle Myocytes; Supervision; System; Targeted Toxins; Techniques; Testing; Tetrachlorodibenzodioxin; Time; Tissues; Toxic Environmental Substances; Toxin; Training; United States National Academy of Sciences; United States National Institutes of Health; Universities; Variant; Xenobiotics; analog; angiogenesis; base; biological research; biological systems; career; cell type; design; efficacy testing; environmental chemical; human disease; improved; in vivo; innovation; instrument; interdisciplinary approach; interest; lectures; lipid mediator; lipophilicity; liquid chromatography mass spectrometry; mass spectrometer; meetings; member; painful neuropathy; personal care products; programs; public health relevance; receptor; response; skills; technical writing; tissue repair; tool; triclocarban

 DESCRIPTION (provided by applicant): The candidate's career goal is to become an independent research investigator and make major contributions to advance science in order to improve human health. The candidate is particularly interested in using chemical probes to understand how environmental toxins affect human health through targeting EET and related lipid-mediators signaling pathways. He has had excellent training in chemical biology, protein chemistry, and molecular biology throughout his career. He has applied his knowledge of using chemical probes to study biological effects to several key systems, including that of cytochrome P450, rhodopsin, and soluble epoxide hydrolase (sEH). He now plans to further complement his training by learning to conduct specialized cell-based assays, advanced proteomics, to operate state-of-the-art accelerator mass spectrometer (AMS), and to analyze AMS data in order to identify the receptor(s) of EETs for the first time. During the K99 phase, he will attend courses and seminars, and will present at national meetings in order to improve his technical writing, presentation, teaching, and leadership skills. By the end of the award, he will have publication records and more than enough preliminary data to apply for R21 and R01 grants through the NIH. During the K99 phase, the candidate will be trained under the supervision of Dr. Bruce Hammock at the University of California, Davis. Dr. Hammock is an elected member of National Academy of Science who studies how environmental exposures affect human health. His research focuses on the xenobiotic enzyme called sEH, a major enzyme that degrades epoxy fatty acids. Dr. Hammock is the pioneer of using a multidisciplinary approach to characterize sEH and to study its role in human diseases like hypertension, inflammation, cancer, fibrosis, and neuropathic pain. Through inhibition of sEH, Dr. Hammock has shown that epoxy fatty acids from ω-6 dietary and ω-3 dietary acids play a vital role in several biological processes such as cardiovascular functioning and inflammation response. In addition, the candidate will learn to run specialized cell-based assays related to the cardiovascular system under co-mentor Dr. Chiamvimonvat who is an expert in cardiovascular disease. He will also learn advance proteomics from Dr. Gomes who also has an excellent track record in this field. Lastly, he will learn to operate AMS and to analyze AMS data under the supervision of Dr. Bruce Buchholz, an expert in AMS. UC Davis is one of the top institutions in the world for biological research. Its programs in environmental science and biological science are internationally acclaimed, ranking twelfth and twenty-fifth in the world, respectively. The institution hosts 800 faculty members who are specialized in biological sciences or biomedical research. The departments at UC Davis organize seminars, lectures, workshops, and discussion sessions on a daily basis, which will allow the candidate to interact with experts in different areas. UC Davis also has many core facilities that house a number of state-of-the-art instruments. These instruments are made easily accessible to scientists at UC Davis and training is always provided. Thus, the candidate can both expand the breadth of his academic knowledge and also learn to use specialized instruments that will help his research. The goal of this proposed project is to identify the receptor(s) of epoxy-eicosanoids, which are lipid signaling molecules. Epoxy-eicosanoids, also known as epoxyeicosatrienoic acids (EETs), are potent chemical mediators that play important roles in inflammation, vasoregulation, analgesia, and angiogenesis. The in vivo levels of EETs are greatly affected by exposure to environmental toxins, such as triclocarban (TCC), 2, 3, 7, 8- tetrachloro-dibenzodioxin (TCDD), and peroxisome proliferators. The changes in in vivo levels of EETs lead to physiological changes that could affect human health. The long-term goal for this project is to understand how the modulation of EET and related lipid mediators signaling pathways affect human health. Although there have been decades of research on this subject, the molecular mechanism of how EETs initiate the signal transduction cascade remains unknown. In the proposed research, the candidate will test the hypothesis that EETs induce the signaling pathway through binding to specific cell-surface receptor(s). The ultimate goal of this proposal is to identify the receptors of EETs for the first time. To identify the receptor(s) of EETs, which are both lipophilic and labile, the intent is to use a combination of C-14 (14C) mass label, photo labels and AMS. AMS, which counts 14C atoms directly, is 100K times more sensitive than any of the traditional decay counting method. This dramatically increases sensitivity has several advantages that will strongly enhance this chances of identifying the EET receptors. The candidate hypothesizes that using AMS with the use of 14C ligands coupled with unique photo labels represents a new method to identify very low-abundance receptors with highly lipophilic and liable ligands. This project, when accomplished, will have a huge impact on basic science and public health. 1) Identification of the receptor(s) of EETs will allow us to bettr understanding the signaling pathway of EETs. 2) The identified EET receptor(s) will provide a way to screen for the environmental toxins that target EET pathway. 3) The method for receptor identification developed in this proposal will become a new method for general receptor identification.

 应聘者描述(申请人提供)：应聘者的职业目标是成为一名独立的研究调查员，并为促进科学进步做出重大贡献，以改善人类健康。这位候选人对使用化学探针了解环境毒素如何通过靶向EET和相关的脂质中介信号通路影响人类健康特别感兴趣。在他的整个职业生涯中，他接受了出色的化学生物学、蛋白质化学和分子生物学方面的培训。他运用他的化学探针知识来研究几个关键系统的生物效应，包括细胞色素P450、视紫红质和可溶性环氧化物水解酶(SEH)。他现在计划通过学习进行专门的基于细胞的分析、高级蛋白质组学、操作最先进的加速器质谱仪(AMS)以及分析AMS数据来进一步补充他的培训，以便首次识别EETs的受体(S)。在K99阶段，他将参加课程和研讨会，并将在全国会议上发言，以提高他的技术写作、演示、教学和领导技能。到颁奖结束时，他将拥有出版记录和足够的初步数据，以通过NIH申请R21和R01拨款。在K99阶段，候选人将在加州大学戴维斯分校的Bruce Hammock博士的监督下接受培训。汉莫克博士是美国国家科学院的当选成员，专门研究环境暴露对人类健康的影响。他的研究重点是一种名为sEH的异生酶，这是一种降解环氧脂肪酸的主要酶。Hammock博士是使用多学科方法来表征sEH并研究其在高血压、炎症、癌症、纤维化和神经病理性疼痛等人类疾病中的作用的先驱。通过抑制sEH，Hammock博士已经证明，来自ω-6膳食和ω-3膳食的环氧脂肪酸在心血管功能和炎症反应等几个生物过程中发挥着至关重要的作用。此外，候选人还将学习在心血管疾病专家、共同导师Chiamvimonvat博士的指导下进行与心血管系统相关的专业细胞检测。他还将向戈麦斯博士学习高级蛋白质组学，戈麦斯博士在这一领域也有出色的记录。最后，他将在AMS专家Bruce Buchholz博士的监督下学习操作AMS并分析AMS数据。加州大学戴维斯分校是世界上最顶尖的生物研究机构之一。中国的环境科学和生物科学项目在国际上享有盛誉，分别位居世界第12位和第25位。该机构拥有800名专门从事生物科学或生物医学研究的教职员工。加州大学戴维斯分校的部门每天都会组织研讨会、讲座、研讨会和讨论会，让应聘者能够与不同领域的专家进行互动。加州大学戴维斯分校还拥有许多核心设施，其中包括许多最先进的乐器。加州大学戴维斯分校的科学家可以很容易地接触到这些仪器，并始终提供培训。因此，考生既可以扩大他的学术知识广度，也可以学习使用有助于他研究的专门工具。这个项目的目标是确定环氧二十烷的受体(S)，它是脂类信号分子。环氧二十碳二烯酸(EETs)又称环氧二十碳三烯酸(EETs)，是一类重要的化学介质，在炎症、血管调节、镇痛和血管生成等方面发挥重要作用。环境毒素，如三氯卡班(TCC)、2，3，7，8-四氯二苯二恶英(TCDD)和过氧化物酶增殖物对体内EETs水平有很大影响。体内EETs水平的变化会导致可能影响人类健康的生理变化。该项目的长期目标是了解EET和相关脂质介质信号通路的调节如何影响人类健康。虽然对这一课题的研究已经有几十年的历史，但EET如何启动信号转导级联的分子机制仍不清楚。在拟议的研究中，候选人将检验EET通过与特定细胞表面受体(S)结合来诱导信号通路的假设。这项提议的最终目标是首次确定EETs的受体。为了鉴定EET的受体(S)，它既亲脂又不稳定，目的是使用C-14(14C)质量标记、照片标记和AMS的组合。AMS直接计算14C原子，其灵敏度是任何传统衰变计数方法的100K倍。这极大地提高了灵敏度，有几个优点将大大增加识别EET受体的机会。候选人假设，使用14C配体结合独特的光标记的AMS代表了一种识别极低丰度受体的新方法，这些受体具有高度亲脂性和易受攻击的配体。该项目一旦完成，将对基础科学和公共卫生产生巨大影响。1)EETs受体(S)的鉴定将有助于我们更好地了解EETs的信号转导途径。2)已鉴定的EET受体(S)将提供一种筛选靶向EET途径的环境毒素的方法。3)本研究提出的受体识别方法将成为一种新的通用受体识别方法。