Airborne PCBs: Sources, Exposures, Toxicities, Remediation: K.C. Donnelly Externship - Promotion of Translational/Transdisciplinary Efforts in Graduate and Post-Doctoral Research

空气中多氯联苯：来源、暴露、毒性、修复：K.C.

• 批准号: 10579012
• 负责人: Keri C Hornbuckle
• 金额: $ 1.55万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579012
• 关键词: Address; Adolescence; Adolescent; Air; Area; Astrocytes; Back; Bacteria; Biochemical; Biological; Blood Circulation; Brain; CYP2B6 gene; CYP3A4 gene; Cell Culture Techniques; Cells; Chemicals; Child; Communication; Communities; Complex; Cytochrome P450; Cytochromes; Data; Data Science; Detection; Development; Environment; Environmental Health; Environmental Pollutants; Enzymes; Evaluation; Exposure to; Family; Freezing; Future; Genes; Glucuronides; Glutathione; Goals; Health; Hepatocyte; Home; Human; In Vitro; Individual; Indoor environment; Industrialization; Inhalation; Inhalation Exposure; Intervention; Iowa; Knowledge; Laboratories; Lead; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolic; Metabolic Biotransformation; Metabolic syndrome; Metabolism; Methods; Modeling; Molecular; Mothers; Neuraxis; Neurodevelopmental Impairment; Oxides; Parents; Personal Satisfaction; Persons; Polychlorinated Biphenyls; Population; Postdoctoral Fellow; Production; Program Research Project Grants; Protein Isoforms; Public Health; Quinones; Rattus; Recommendation; Research; Research Activity; Research Personnel; Research Priority; Research Project Grants; Resolution; Risk; Risk Assessment; Risk Factors; Role; Route; Sampling; Schools; Scientist; Site; Soil; Source; Sulfate; Superfund; Testing; Toxic effect; Toxicokinetics; Toxicology; Training; Training Programs; Translating; Translational Research; Universities; Work; Xenobiotics; adduct; base; building materials; career; cohort; community engagement; contaminated sediment; contaminated water; cost effective; design; detection method; dietary; experience; exposed human population; externship; fungus; improved; innovation; lipid metabolism; microbial; neurodevelopment; new technology; novel; novel strategies; overexpression; oxidation; prevent; programs; remediation; skills; superfund chemical; superfund site; tool; translational study

PROJECT DESCRIPTION Due to inadvertent production and exposure to polychlorinated biphenyls (PCBs) from environmental sources, adverse health effects remain a critical public health issue long after the ban of commercial production. Through the proposed research, our goal is to gain knowledge related to P450-mediated metabolism through the KC Donnelly externship in partnership with the National Center for Toxicology Research (NCTR). Xenobiotic processing genes (XPGs), such as cytochrome P450s, are known to be expressed and active in the brain [1-8]. This can then lead to the metabolism of central nervous system (CNS)-active chemicals, leading to alterations in toxicokinetics and toxicodynamics of target cells through systemic circulation [9, 10]. Currently, I am working on examining metabolism of PCBs in rat primary astrocytes. However, it is still uncertain where metabolism of PCBs begins. Specifically, following ISRP Project 1, Aim 2, I will be examining region-specific biotransformation of PCBs and PCB metabolites in the adolescent rat brain in vitro (primary cells in culture) to characterize the following local metabolic conversions: oxidation of parent PCBs to OH-PCBs; conjugation of OH- and diOH-PCBs; deconjugation of OH-PCB sulfates (and glucuronides); and oxidation of diOH-PCBs to reactive quinones and subsequently, glutathione adducts (reviewed in [11-13]). To expand upon this work, the precise human metabolism of lower chlorinated PCBs is still uncertain and needs to be explored.Therefore, my overall objective is to examine the P450 isoform-specific metabolism of lower-chlorinated PCBs in state-of-the- art P450-overexpressing human hepatocytes developed by Dr. Lei Guo's laboratory at the National Center for Toxicology Research (NCTR) [14]. This model will be used to test the hypothesis that lower-chlorinated PCBs discovered by the ISRP in indoor and outdoor air, such as 2,2'5,5'-tetrachlorobiphenyl (PCB52) and 3,3’- dichlorobiphenyl (PCB11), are oxidized with different stereoselectivity by several human P450 isoforms, such as CYP2A6, CYP2B6, and CYP3A4 I will test this hypothesis by (1) determining which human P450 isoform is involved in the metabolism of PCB11 and PCB52 and (2) characterizing the PCB metabolites formed by different P450 isoforms using nontarget-high resolution mass spectrometry (Nt-HRMS). These results will inform future translational studies that assess the role of these P450 isoforms in the brain specific metabolism in human-relevant cell culture models. After treating the P450-overexpressing hepatocytes, both cell pellets and supernatant fractions will be collected to assess levels of metabolites present. Supernatant and cells will be flash frozen. These samples would then be transported back to the University of Iowa, where subsequent quantification of key metabolites would be accomplished with standards provided by the ISRP Synthesis Core [15-19]. The proposed studies will investigate the P450 specific metabolism of PCBs in hepatocytes and are a logical extension of the current ISRP goals to determine enzyme-specific metabolism.

项目描述 由于无意的产生和暴露于环境来源的多氯联苯（PCB）， 在禁止商业生产后很长时间，不利的健康影响仍然是一个关键的公共卫生问题。 通过拟议的研究，我们的目标是通过 KC Donnelly与国家毒理学研究中心（NCTR）合作。 已知在 大脑[1-8]。然后，这可能导致中枢神经系统（CNS）活性化学物质的代谢，导致 靶细胞通过全身循环的毒性动力学和毒性动力学的改变[9，10]。目前，我 正在研究大鼠原发性星形胶质细胞中PCB的代谢。但是，仍然不确定在哪里 PCB的代谢开始。具体而言，以下ISRP项目1，AIM 2，我将检查特定区域 PCB和PCB代谢物在青少年大鼠脑（培养中的原代细胞）中的生物转化 表征以下局部代谢转化率：将父型PCB氧化为OH-PCB；共轭 哦，dioh-pcb; OH-PCBS（和葡萄糖醛酸苷）的解偶；和DiOH-PCB的氧化 反应性奎因酮，随后是谷胱甘肽加合物（在[11-13]中进行了回顾）。为了扩大这项工作， 下层氯化PCB的精确人类代谢仍然不确定，需要探索。因此，我 总体目的是检查较低氯化PCB的P450同工型特异性代谢 由国家中心的Lei Guo博士的实验室开发的ART P450过表达的人类肝细胞 毒理学研究（NCTR）[14]。该模型将用于测试下氯的PCB的假设 由ISRP在室内和室外空气中发现，例如2,2'5'-tetrachlorobiphenyl（PCB52）和3,3'-- 二氯贝苯基（PCB11）被几种人P450同工型氧化，以不同的立体选择性氧化 如CYP2A6，CYP2B6和CYP3A4，我将通过（1）确定人类P450同工型是 参与PCB11和PCB52的代谢和（2）表征由PCB代谢产生的（2） 使用非目标高分辨率质谱法（NT-HRMS）的不同P450同工型。这些结果将会 告知未来的翻译研究，这些研究评估这些P450同工型在大脑特定代谢中的作用 在与人相关的细胞培养模型中。处理P450过表达的肝细胞后，两个细胞颗粒 将收集上清液，以评估存在的代谢水平。上清液和细胞将 闪烁冷冻。然后将这些样本运回爱荷华大学，随后到达那里 通过ISRP合成核提供的标准，将完成数量的关键代谢物 [15-19]。拟议的研究将研究肝细胞中PCB的P450特异性代谢，并且是一个 当前ISRP目标的逻辑扩展，以确定酶特异性代谢。