Exposure and biological response biomarkers of cigarette smoke

香烟烟雾的暴露和生物反应生物标志物。暴露在烟草烟雾中 (主流和环境)是美国主要的死亡原因。香烟烟雾是一种极端的 复杂的混合物，约3800种成分，包括许多多环芳烃 (PAHs)，在主流和侧流(环境)烟雾组分中。吸烟者提供 多环芳烃暴露的极端模型将允许暴露和生物反应生物标记物 发展起来的。有大量证据表明，多环芳烃是肺癌、皮肤癌和膀胱癌的病原体。 此外，烟草烟雾还与氧化应激、胰腺癌、心血管疾病、 和慢性阻塞性肺疾病(COPD)，尽管多环芳烃的具体作用尚不清楚。 有趣的是，侧流烟雾对心血管的影响几乎与主流烟雾一样大。 目前的建议源于我们在过去六年中取得的重大进展 利用稳定同位素方法和我们的基础研究对蛋白质、脂肪和DNA生物标志物进行定量 氧化应激过程中的酶调节。氧化型DNA分析的几种常用方法 损害一直充斥着无数的方法问题，因此目前最先进的 涉及使用彗星试验来测量8-氧-2‘-脱氧鸟苷(Dguo)损伤。我们最近做了 设计了一种基于免疫亲和稳定同位素稀释液相色谱的更定量的方法- 串联质谱仪(LC-MS/MS)，可以很容易地阐述到烟草吸烟者的研究。我们 也表明氧化应激可以诱导1C的醛酮还原酶(AKRs)的形成 一家人。AKR1C3是一种酶，我们最近发现它负责前列腺素的转化 (Pg)D2至强效的支气管收缩因子11P-PGF2。这提供了一个额外的潜在联系 氧化应激和COPD以及新治疗策略的可能性，其中包括AKR1C3 抑制力。最后，初步研究表明，DNA加合物只能由脂质产生 吸烟者的尿液中存在过氧化作用，而不吸烟者的尿液中则完全没有过氧化反应。 我们建议在这些令人兴奋的新发现的基础上，开发暴露在体内的生物标记物小组。 和生物反应，我们假设这将使区分一群不吸烟的人成为可能 来自一群没有疾病的烟草吸烟者。这一假说将通过以下方式进行研究来检验 目的1.探讨B[a]P和B[a]P-7，8-二酮是否诱导AKR1C/2 并增加氧化应激在DNA中形成8-oxo-dGuo和HedGuo，诱导AKR1C3在 HASM细胞，增加强大的支气管收缩因子11P-PGF2的生物合成，作为潜在的尿液和 多环芳烃暴露的EEC生物反应生物标志物。目标2：发现下列分泌蛋白 苯并[a]P及其氧化代谢产物作为潜在血清生物制剂处理NHBE和HASM细胞 多环芳烃暴露的反应生物标志物。目标3：进行体内预测和改进分析 尿中暴露和反应生物标志物以及EBC和血清中的生物反应生物标志物 以区分不吸烟者和无疾病吸烟者。圆满完成 拟议的研究将提供一组暴露和对烟草烟雾的生物反应的生物标记物 将在未来旨在阐明基因环境之间关系的研究中具有重要的实用价值 相互作用和疾病，如癌症、心血管疾病和慢性阻塞性肺病。

Exposure and biological response biomarkers of cigarette smoke. Exposure to tobacco smoke (mainstream and environmental) is a leading cause of death in the US. Cigarette smoke is an extremely complex mixture, which some 3800 constituents including numerous polycyclic aromatic hydrocarbons (PAHs), in both the mainstream and sidestream (environmental) smoke fractions. Cigarette smokers provide an extreme model of PAH exposure that will permit both exposure and biological response biomarkers to be developed. There is substantial evidence that PAHs are causative agents in lung, skin, and bladder cancer. Furthermore, tobacco smoke is associated with oxidative stress, pancreatic cancer, cardiovascular disease, and chronic obstructive pulmonary disease (COPD), although the specific role of PAHs is not clear. Interestingly, the cardiovascular effects of sidestream smoke are almost as great as mainstream smoke. The present proposal stems from significant advances we have made over the last six years in the quantification of protein, lipid, and DNA biomarkers using stable isotope methodology and our basic research into enzyme regulation during oxidative stress. Previous methods for he the analysis of oxidative DNA damage has been fraught with numerous methodological problems so that the current state-of-the-art involves the use of a COMET assay to measure 8-oxo-2'-deoxyguanosine (dGuo) lesions. We have recently devised a more quantitative method based on immunoaffinity stable isotope dilution liquid chromatography- tandem mass spectrometry (LC-MS/MS) that can be readily elaborated to studies of tobacco smokers. We also showed that oxidative stress could induce the formation of aldo-keto reductases (AKRs) of the 1C family. AKR1C3 is the enzyme, which we recently showed is responsible for the conversion of prostaglandin (PG) D2 to the potent bronchoconstrictor 11p-PGF2. This provides an additional potential link between oxidative stress and COPD as well as the potential for a new therapeutic strategy, which involves AKR1C3 inhibition. Finally, preliminary studies have revealed that a DNA-adduct than can only arise from lipid peroxidation is present in the urine of cigarette smokers but is completely absent in urine from non-smokers. We propose to build on these exciting new findings by developing panels of in vivo biomarkers of exposure and biological response, which we hypothesize will make it possible to distinguish a cohort of non-smokers from a cohort of disease-free tobacco smokers. The hypothesis will be tested by conducting research under the following three specific aims: Aim 1. To discover whether B[a]P and B[a]P-7,8-dione induce AKR1C/2 in NHBE cells and increase oxidative stress to form 8-oxo-dGuo and HedGuo in DNA, induce AKR1C3 in HASM cells and increase the biosynthesis of the potent bronchoconstrictor 11p-PGF2, as potential urine and EEC biological response biomarkers of PAH exposure. Aim 2: To discover secreted proteins following treatment of NHBE and HASM cells with B[a]P and its oxidative metabolites as potential serum biological response biomarkers of PAH exposure. Aim 3: To conduct predictive and refinement analyses of in vivo exposure and response biomarkers in urine together with biological response biomarkers in EBC and serum in order to distinguish non-smokers from disease-free tobacco smokers. Successful completion of the proposed research will provide a panel of biomarkers of exposure and biological response to tobacco smoke will have significant utility in future studies designed to elucidate the relationship between gene environment interactions and diseases such as cancer, cardiovasculardisease, and COPD.