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: 8977513
• 负责人: Kin Sing Stephen Lee
• 金额: $ 6.49万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2016-12-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8977513
• 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; Housing; Human; Hypertension; In Vitro; Individual; Inflammation; Institution; 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; Pain; Pathway interactions; 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; Techniques; Testing; Time; Tissues; Toxic Environmental Substances; 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; liquid chromatography mass spectrometry; mass spectrometer; meetings; member; painful neuropathy; personal care products; programs; protein aminoacid sequence; radiotracer; 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.

 描述（由申请人提供）：候选人的职业目标是成为一名独立的研究人员，为推动科学进步以改善人类健康做出重大贡献。该候选人特别感兴趣的是使用化学探针来了解环境毒素如何通过靶向 EET 和相关脂质介质信号通路来影响人类健康。在他的职业生涯中，他在化学生物学、蛋白质化学和分子生物学方面接受过良好的培训。他将使用化学探针的知识应用于几个关键系统研究生物效应，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测定、先进的蛋白质组学、操作最先进的加速器质谱仪 (AMS) 以及分析 AMS 数据以首次识别 EET 的受体来进一步补充他的培训。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。到获奖结束时，他将拥有发表记录和足够多的初步数据，可以通过 NIH 申请 R21 和 R01 资助。在K99阶段，候选人将在加州大学戴维斯分校Bruce Hammock博士的监督下接受培训。 Hammock 博士当选为美国国家科学院院士，研究环境暴露如何影响人类健康。他的研究重点是称为 sEH 的外源酶，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、纤维化和神经性疼痛等人类疾病中的作用的先驱。 Hammock 博士通过抑制 sEH 证明，来自 ω-6 膳食酸和 ω-3 膳食酸的环氧脂肪酸在心血管功能和炎症反应等多种生物过程中发挥着至关重要的作用。此外，候选人还将在心血管疾病专家 Chiamvimonvat 博士的指导下学习运行与心血管系统相关的专门细胞检测。他还将向戈麦斯博士学习先进的蛋白质组学，戈麦斯博士在该领域也拥有出色的记录。最后，他将在AMS专家Bruce Buchholz博士的指导下学习操作AMS并分析AMS数据。加州大学戴维斯分校是世界顶尖的生物研究机构之一。其环境科学和生物科学项目享誉国际，分别排名世界第十二和二十五。该机构拥有 800 名专门从事生物科学或生物医学研究的教员。加州大学戴维斯分校的各院系每天都会组织研讨会、讲座、讲习班和讨论会，这将使候选人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，配备了许多最先进的仪器。加州大学戴维斯分校的科学家可以轻松使用这些仪器，并且始终提供培训。因此，候选人既可以扩大学术知识的广度，又可以学习使用有助于他的研究的专业仪器。该项目的目标是识别环氧类二十烷酸的受体，它们是脂质信号分子。环氧二十烷酸，也称为环氧二十碳三烯酸 (EET)，是有效的化学介质，在炎症、血管调节、镇痛和血管生成中发挥重要作用。 EET 的体内水平受到环境毒素暴露的极大影响，例如三氯卡班 (TCC)、2,3,7,8-四氯二苯并二恶英 (TCDD) 和过氧化物酶体增殖剂。 EET 体内水平的变化会导致生理变化，从而影响人类健康。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。尽管对这个课题已经进行了数十年的研究，但 EET 如何启动信号转导级联的分子机制仍然未知。在拟议的研究中，候选人将测试 EET 通过与特定细胞表面受体结合诱导信号传导途径的假设。该提案的最终目标是首次鉴定EETs的受体。为了识别亲脂性且不稳定的 EET 受体，目的是结合使用 C-14 (14C) 质量标签、照片标签和 AMS。 AMS 直接计数 14C 原子，其灵敏度比任何传统的衰变计数方法高 100K 倍。这显着提高了灵敏度，有几个优点，将大大提高识别 EET 受体的机会。候选者假设，使用 AMS 结合 14C 配体和独特的照片标签代表了一种识别具有高亲脂性和可靠配体的极低丰度受体的新方法。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受体的鉴定将使我们能够更好地了解EETs的信号通路。 2) 已鉴定的 EET 受体将提供一种筛选针对 EET 途径的环境毒素的方法。 3）本提案开发的受体鉴定方法将成为通用受体鉴定的新方法。

项目成果

Soluble epoxide hydrolase inhibitor 1-trifluoromethoxyphenyl-3- (1-propionylpiperidin-4-yl) urea attenuates bleomycin-induced pulmonary fibrosis in mice.

• DOI: 10.1007/s00441-015-2262-0
• 发表时间: 2016-02
• 期刊: Cell and tissue research
• 影响因子: 3.6
• 作者: Zhou Y;Yang J;Sun GY;Liu T;Duan JX;Zhou HF;Lee KS;Hammock BD;Fang X;Jiang JX;Guan CX
• 通讯作者: Guan CX