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: 8805755
• 负责人: Kin Sing Stephen Lee
• 金额: $ 6.68万
• 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/8805755
• 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; Self Care; 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; programs; protein aminoacid sequence; public health relevance; 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（由适用提供）：候选人的职业目标是成为一名独立的研究研究者，并为进步科学做出重大贡献，以改善人类健康。候选人特别有兴趣使用化学问题来了解环境毒素如何通过靶向EET和相关脂质中的信号通路来影响人类健康。他在整个职业生涯中都接受了化学生物学，蛋白质化学和分子生物学的出色培训。他已经应用了使用化学问题来研究生物学效应的知识，包括几个关键系统，包括细胞色素P450，视紫红质和固体环氧化物水解酶（SEH）。现在，他计划通过学习进行专业的基于细胞的测定方法，高级蛋白质组学，以进一步补充他的培训，以运行最先进的加速器质谱仪（AMS），并分析AMS数据，以便第一次识别EET的接收者。在K99阶段，他将参加课程和准则，并将在全国会议上举行，以提高他的技术写作，演讲，教学和领导能力。颁奖典礼结束时，他将拥有出版记录和足够的初步数据，可以通过NIH申请R21和R01赠款。在K99阶段，候选人将在加利福尼亚大学戴维斯分校的布鲁斯·汉莫克博士的监督下进行培训。 Hammock博士是国家科学院当选成员，研究环境暴露如何影响人类健康。他的研究重点是称为SEH的异生元，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，感染，癌症，纤维化和神经性疼痛等人类疾病中的作用的先驱。通过抑制SEH，Hammock博士表明，来自ω-6饮食和ω-3饮食酸的环氧脂肪酸在多种生物学过程中起着至关重要的作用，例如心血管功能和炎症反应。此外，候选人将学会在心血管疾病专家Chiamvimonvat博士的下经营与心血管系统有关的专门基于细胞的麻烦。他还将从Gomes博士那里学习进步蛋白质组学，后者在该领域也有出色的往绩。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重要的生物学研究机构之一。它在环境科学和生物科学方面的计划在国际上广受好评，在世界范围内分别排名第十二和二十五。该机构拥有800名专门从事生物科学或生物医学研究的教职员工。加州大学戴维斯分校的部门每天组织半手，讲座，讲习班和讨论会议，这将使候选人能够与不同领域的专家互动。加州大学戴维斯分校还拥有许多核心设施，可容纳许多最先进的工具。加州大学戴维斯分校的科学家可以轻松访问这些乐器，并始终提供培训。这是候选人都可以扩大他的学术知识的广度，还可以学会使用有助于他研究的专业工具。该提议的项目的目的是识别环氧树脂 - 透氧烷的接收器，即脂质信号分子。环氧树脂 - 也称为环氧树钠酸（EET），是潜在的化学介质，它们在炎症，血管调节，镇痛和血管生成中起着重要作用。体内EET的水平受到环境毒素的暴露很大的影响，例如Triclocarban（TCC），2、3、7、8- tetrachloro-dibenzodioxin（TCDD）和过氧化物组扩散剂。体内EET水平的变化导致身体变化，可能会影响人类健康。该项目的长期目标是了解EET和相关脂质介质的调节如何影响人类健康。尽管已经对该主题进行了数十年的研究，但EET启动信号转导级联的分子机制仍然未知。在拟议的研究中，候选人将检验以下假设，即EET通过与特定细胞表面受体的结合诱导信号通路。该提案的最终目标是首次识别EET的受体。为了识别既是亲脂性和标签的Eets的受体，意图是使用C-14（14C）质量标签，照片标签和AMS的组合。直接计数14c原子的AMS比任何传统的衰减计数方法都高100k倍。这显着提高了灵敏度具有多种优势，可以强烈增强这种识别EET受体的机会。候选人假设使用AMS与14C配体配合使用独特的照片标签代表了一种新的方法，可以识别具有高亲脂性和责任配体的非常低的丰度受体。该项目完成后，将对基础科学和公共卫生产生巨大影响。 1）鉴定EET的受体将使我们能够理解EET的信号传导途径。 2）鉴定的EET受体将提供一种筛选靶向EET途径的环境毒素的方法。 3）在本提案中开发的受体鉴定方法将成为一种通用受体识别的新方法。