Identifying the Receptors of Environmentally Sensitive Epoxy-Eicosanoids with AMS

 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调查员，并为促进科学做出重大贡献，以改善人类健康。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和相关的脂质介体信号通路影响人类健康。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了出色的培训。他运用化学探针的知识研究了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测定，先进的蛋白质组学，操作最先进的加速器质谱仪（AMS），并分析AMS数据，以首次识别Ehrs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，并将出席国家会议，以提高他的技术写作，演讲，教学和领导技能。到获奖结束时，他将有出版记录和足够的初步数据通过NIH申请R21和R 01赠款。在K99阶段，候选人将在加州大学戴维斯分校布鲁斯吊床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，研究环境暴露如何影响人类健康。他的研究重点是一种名为sEH的异生素酶，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。通过抑制sEH，Hammock博士已经表明，ω-6膳食酸和ω-3膳食酸中的环氧脂肪酸在几个生物过程中起着至关重要的作用，如心血管功能和炎症反应。此外，候选人将学习在心血管疾病专家Chiamvimonvat博士的指导下运行与心血管系统相关的专门细胞检测。他还将从Gomes博士那里学习先进的蛋白质组学，Gomes博士在这一领域也有出色的记录。最后，他将在AMS专家布鲁斯布赫霍尔茨博士的指导下学习操作AMS和分析AMS数据。加州大学戴维斯分校是世界上生物研究的顶尖机构之一。它在环境科学和生物科学方面的课程在国际上享有盛誉，分别排名世界第十二和第二十五位。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。UC Davis的各部门每天都会组织研讨会，讲座，研讨会和讨论会，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，其中包括许多最先进的仪器。加州大学戴维斯分校的科学家们很容易使用这些仪器，并始终提供培训。因此，候选人既可以扩大他的学术知识的广度，也可以学习使用专业仪器，这将有助于他的研究。该项目的目标是鉴定环氧类花生酸的受体，这是脂质信号分子。环氧-类二十烷酸，也称为环氧二十碳三烯酸（ECONOXYLENE），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。暴露于环境毒素，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，极大地影响了体内的雌二醇水平。体内雌二醇水平的变化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。虽然已经有几十年的研究在这个问题上，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Ehrs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确定Escherichia coli的受体。为了鉴定亲脂性和不稳定性的E14受体，目的是使用C-14（14 C）质量标记、照片标记和AMS的组合。AMS直接对14 C原子进行计数，比传统的衰变计数方法灵敏10万倍。这大大增加了灵敏度，具有几个优点，将大大提高识别EET受体的机会。候选人假设，使用AMS与14 C配体结合使用独特的光标记代表了一种新的方法，以确定非常低丰度的受体与高度亲脂性和可靠的配体。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)E2受体的鉴定将有助于我们更好地了解E2的信号通路。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)本研究建立的受体鉴定方法将成为一种新的受体鉴定方法。

 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.