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，为推动科学发展做出重大贡献，以改善人类健康。候选人特别有兴趣使用化学探针来了解环境毒素如何影响人类健康。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的生物效应研究。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) 来进一步补充他的培训。分析AMS数据，以便首次识别EET的受体。在K99阶段，他将参加课程和研讨会，并将按顺序出席国家会议。提高他的技术写作、演讲、教学和领导技能。在获奖结束时，他将拥有足够的发表记录和足够的初步数据，以通过 NIH 申请 R21 和 R01 资助。将在加州大学戴维斯分校的 Bruce Hammock 博士的监督下接受培训。Hammock 博士是美国国家科学院的当选成员，他的研究重点是称为 sEH 的外源酶。一个Hammock 博士是使用多学科方法来表征 sEH 并通过抑制 sEH 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 表明，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，考生还将学习如何进行专门的跑步。在心血管疾病专家 Chiamvimonvat 博士的指导下，他还将学习与心血管系统相关的细胞分析。Gomes 博士在该领域也拥有出色的记录。在AMS专家Bruce Buchholz博士的监督下操作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在国际上享有盛誉。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，每天都会组织研讨会、讲座、讲习班和讨论会。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，这些仪器可供加州大学戴维斯分校的科学家轻松使用，并且始终提供培训。因此，候选人既可以扩展其学术知识的广度，又可以学习使用有助于其研究的专用仪器。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），是一种有效的化学介质，在炎症、血管调节、镇痛和血管生成中发挥重要作用。 EET 的含量受环境毒素的影响很大，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。体内 EET 水平的变化会导致生理变化。该项目的长期目标是了解 EET 和脂质相关介质信号通路的调节如何影响人类健康。在本课题中，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。该提案是首次鉴定 EET 受体 为了鉴定亲脂性和不稳定的 EET 受体，目的是使用 C-14 (14C) 质量标签、照片标签的组合。 AMS 直接对 14C 原子进行强计数，其灵敏度比任何传统的衰变计数方法高 100K 倍，这具有多个优点，将提高识别 EET 受体的机会。使用 14C 配体结合独特的照片标签代表了一种识别具有高亲脂性和敏感配体的极低丰度受体的新方法。该项目一旦完成，将对基础科学和公众产生巨大影响。 1) EET 受体的鉴定将使我们能够更好地了解 EET 的信号传导途径。 2) 鉴定出的 EET 受体将提供一种筛选针对 EET 途径的环境毒素的方法。 ）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.

可溶性环氧化物水解酶抑制剂 1-三氟甲氧基苯基-3-(1-丙酰哌啶-4-基)尿素可减轻博莱霉素诱导的小鼠肺纤维化。

• 发表时间: 2016-02
• 影响因子: 3.6
• 作者: Zhou, Yong;Yang, Jun;Sun, Guo;Liu, Tian;Duan, Jia;Zhou, Hui;Lee, Kin Sing;Hammock, Bruce D;Fang, Xiang;Jiang, Jian;Guan, Cha
• 通讯作者: Guan, Cha

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

勘误：可溶性环氧化物水解酶抑制剂 1-三氟甲氧基苯基-3-(1-丙酰哌啶-4-基) 尿素可减轻博莱霉素诱导的小鼠肺纤维化。

• 发表时间: 2016-06
• 影响因子: 3.6
• 作者: Zhou, Yong;Yang, Jun;Sun, Guo;Liu, Tian;Duan, Jia;Zhou, Hui;Lee, Kin Sing;Hammock, Bruce D;Fang, Xiang;Jiang, Jian;Guan, Cha
• 通讯作者: Guan, Cha