Characterization of Naphthalene DNA Adducts in Mice and Firefighters

小鼠和消防员中萘 DNA 加合物的表征

• 批准号: 10462039
• 负责人: Sarrah Hannon
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10462039
• 关键词: Academia; Adult; Aromatic Polycyclic Hydrocarbons; Aryl Hydrocarbon Hydroxylases; Benzo(a)pyrene; Biological Assay; Blood; Blood specimen; Carcinogens; Career Mobility; Cells; Collaborations; Cytochrome P450; DNA; DNA Adduction; DNA Adducts; Data; Detection; Development; Environment; Environmental Pollution; Enzymes; Epoxy Compounds; Exposure to; Fellowship; Female; Fire - disasters; Fossil Fuels; Foundations; Future; General Population; Generations; Genes; Human; In Vitro; Incidence; Inhalation; Inhalation Exposure; International Agency for Research on Cancer; Knowledge; Lead; Leukocytes; Literature; Liver; Lung; Malignant Neoplasms; Mass Spectrum Analysis; Metabolic Pathway; Metabolism; Methods; Microsomal Epoxide Hydrolase; Mus; Mutagenesis; NMR Spectroscopy; Naphthalene; Naphthoquinones; Nasal Epithelium; Neuroblastoma; Occupational Groups; Occupations; Oxidoreductase; Pathogenesis; Predictive Value; Public Health; Publishing; Quinones; Rattus; Research; Research Personnel; Risk; Risk Assessment; Site; Smoke; Source; Structure of parenchyma of lung; Testing; Time; Tissues; Tobacco; Toxicokinetics; Training; Transgenic Mice; Urine; Wild Type Mouse; Work; adduct; adenoma; cancer risk; cancer type; carcinogenesis; carcinogenicity; cytotoxic; exposed human population; fire fighter; genetic manipulation; genotoxicity; in vivo; lung carcinogenesis; male; mouse model; professional atmosphere; sex; skills; tumor

Project Summary/Abstract Naphthalene (NA), a simple polycyclic aromatic hydrocarbon, is persistently present in the environment as a byproduct of combustion of fossil fuels and burning of tobacco and other products. Its ubiquitous presence results in widespread exposure to the general population. Certain occupational groups, such as firefighters, have elevated levels of exposure. Firefighters also have increased incidences of certain types of cancer. NA is currently classified by the International Agency for Research on Cancer (IARC) as a Class 2B Carcinogen. There is direct evidence of tumor formation in mice and rats but no direct evidence of carcinogenecity of NA in humans at this time. The pathogenesis of tumor formation in mice and rats after exposure to NA is unclear; cytotoxic and genotoxic mechanisms are both proposed in the current literature. NA metabolism results in the generation of reactive intermediates such as 1,2-epoxide and reactive metabolites such as 1,4- and 1,2- naphthoquinone (NAQ). Reactive quinone and epoxide metabolites of similar compounds, such as benzo[a]pyrene, have been shown to enact their carcinogenecity through DNA adduct formation. Published ex vivo and in vitro data have demonstrated that NA metabolites can form adducts with DNA. The objective of this study is to identify and quantify NA-DNA adducts in mouse lung as well as mouse and human blood to enable assessment of potential genotoxicity in firefighters. Several hypotheses will be tested: 1) Circulating reactive NA metabolites, particularly NAQs, will form stable as well as depurinating adducts with DNA in both lung tissue and blood leukocytes in vivo; 2) the types and abundances of NA-DNA adducts in circulating leukocytes will at least partly reflect adduct formation in the lung; 3) the types of NA-DNA adducts detected in circulating leukocytes from exposed mice and firefighters will be similar; and 4) exposure to fire smoke will increase NA- DNA adduct levels in firefighters. These hypotheses will be tested through the application of 3 Specific Aims. In Aim 1, I will Identify and quantitate NA-DNA adducts formed in wild-type (WT) mice following NA inhalation exposure, with use of mass spectrometry methods. In Aim 2, I will dissect metabolic pathways responsible for NA-DNA adduct formation in the lung and circulating leukocytes of NA-exposed mice, with use of unique transgenic mouse models (liver-Cpr-null, liver-Ephx1-null, and Cyp2abfgs-null). In Aim 3, I will characterize NA-DNA adducts in blood specimens from firefighters and control donors. This project will provide direct evidence for the formation of NA-DNA adducts in vivo, lay the foundation for future studies of the genotoxicity of NA, obtain evidence to support more extensive assessments of the carcinogenic risks of NA to firefighters, and could have direct implications for the general population. Through the execution of this project and the help of this fellowship, I will be fully trained in scientific and professional skills necessary for my career advancement as an independent researcher in academia.

项目摘要/摘要 萘(NA)是一种简单的多环芳烃，在环境中以 燃烧化石燃料以及燃烧烟草和其他产品的副产品。它无处不在的存在 导致对普通人群的广泛接触。某些职业类别，如消防员， 有更高的暴露水平。消防员也增加了某些类型癌症的发病率。那就是 目前被国际癌症研究机构(IARC)列为2B类致癌物。 有直接证据表明小鼠和大鼠体内有肿瘤形成，但没有直接证据表明NA在小鼠和大鼠体内具有致癌作用 人类在这个时候。小鼠和大鼠暴露于NA后肿瘤形成的机制尚不清楚； 在目前的文献中，细胞毒性和遗传毒性机制都被提出。NA新陈代谢导致 生成活性中间体如1，2-环氧化物和活性代谢物如1，4-和1，2- 萘酚(NAQ)。类似化合物的反应性苯二酚和环氧化物代谢物，如 苯并[a]芘，已被证明通过DNA加合物的形成而致癌。已出版的EX 体内和体外数据表明，NA代谢物可以与DNA形成加合物。这样做的目的是 研究是鉴定和定量小鼠肺以及小鼠和人血中的NA-DNA加合物，以使 消防员潜在遗传毒性的评估。将检验几个假设：1)循环反应性 NA代谢物，特别是NAQs，将在两个肺中与DNA形成稳定的和净化的加合物。 体内组织和血白细胞；2)循环白细胞中NA-DNA加合物的类型和丰度 将至少部分反映肺中加合物的形成；3)在循环中检测到的NA-DNA加合物的类型 暴露在烟雾中的小鼠和消防员的白细胞将相似；4)暴露在火灾烟雾中将增加NA- 消防员体内的DNA加合物水平。这些假设将通过三个具体目标的应用来检验。在……里面 目的1.鉴定和定量吸入NA后野生型(WT)小鼠体内形成的NA-DNA加合物 使用质谱学方法进行曝光。在目标2中，我将剖析负责的代谢途径 NA染毒小鼠肺和循环白细胞中NA-DNA加合物的形成 转基因小鼠模型(肝脏CPR-空、肝-Ephx1-空和Cyp2abfgs-空)。在《目标3》中，我将描述 消防员和对照献血者血液样本中的NA-DNA加合物。该项目将直接提供 为在体内形成NA-DNA加合物的证据，为今后的遗传毒性研究奠定了基础 获取证据支持对NA对消防员致癌风险的更广泛评估， 并可能对普通民众产生直接影响。通过执行这一项目和 在此奖学金的帮助下，我将获得职业生涯所需的科学和专业技能方面的全面培训 在学术界晋升为独立研究员。