PROTEIN ADDUCTS AS MOLECULAR SIGNATURES OF CARCINOGEN DOSE

蛋白质加合物作为致癌剂量的分子特征

• 批准号: 8102121
• 负责人: Stephen Morris Rappaport
• 金额: $ 47.8万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8102121
• 关键词: Affinity; Aromatic Polycyclic Hydrocarbons; Arts; Bioinformatics; Biological Assay; Biological Markers; Blood; Blood Proteins; Blood specimen; Bovine Serum Albumin; Carcinogen exposure; Carcinogens; Chemicals; Chronic Disease; Cysteine; Data; Dissociation; Dose; Drops; Environmental Exposure; Enzyme-Linked Immunosorbent Assay; Epidemiology; Exposure to; Fourier transform ion cyclotron resonance; General Population; Hematopoietic Neoplasms; Hemoglobin; Human; Human Volunteers; Investigation; Lasers; Libraries; Link; Longevity; Lymphoma; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measurement; Measures; Methods; Molecular Profiling; Needles; Occupational; Peptides; Plague; Plant Resins; Poison; Population; Proteins; Proteome; Proteomics; Reaction; Reference Standards; Resolution; Review Literature; Route; Sample Size; Sampling; Serum; Serum Albumin; Site; Source; Spottings; Sulfhydryl Compounds; Surface; Time; Tissues; Toxicogenomics; Venous blood sampling; adduct; base; case control; epidemiology study; in vivo; indium arsenide; micro-total analysis system; repaired; sensor; volunteer

Because exposures to carcinogens arise from many sources and routes, epidemiology studies should use biomarkers of internal dose as surrogates for carcinogen exposures. However, direct measurement of carcinogens is impractical because these chemicals are usually reactive electrophiles with short life spans in the body. As an alternative, carcinogen doses can be investigated via addition products (adducts) of the electrophiles with blood proteins, especially human serum albumin (HSA) and hemoglobin (Hb). Because these adducts are not repaired, they reflect doses of carcinogens over one to two months. In this project, we envision a 'protein adductome', representing all adducts present on a given protein, as an entity of toxicogenomic importance. Since all electrophilic species are potentially carcinogenic, the protein adductome is arguably more relevant to carcinogen discovery than the proteome or the metabolome. Despite their utility as biomarkers of carcinogen dose, adducts of toxic chemicals with abundant blood proteins, such as human serum albumin (HSA) and Hb, have rarely been used in epidemiology studies or to discover the initiators of human cancers. There are three major reasons for this. First, without substantial adduct enrichment, typical concentrations of HSA and Hb adducts are too low to permit widespread * discovery of new carcinogens. This is the 'needle-in-a-haystack problem' that has also plagued proteomic investigations. Second, state-of-the-art mass spectrometry (MS) has not been exploited to identify unknown protein adducts or to profile adducts with links to human cancers. And third, it is difficult to obtain blood samples with which to assay protein adducts. In this project, we hypothesize that particular subadductomes, such as the free cysteine of HSA (Cys34), can be used to discover new carcinogens and to serve as biomarkers of internal dose in large epidemiology studies. To consider these questions, we will selectively enrich cysteinyl adducts of HSA from human blood and then will use MS to characterize the known and unknown HSA adducts. By comparing adduct maps between lymphoma cases and control subjects, we will pinpoint possible carcinogens. Regarding the use of protein adducts in epidemiology studies, we will collaborate with Project 3 to determine whether it is feasible to quickly measure protein adducts of polycyclic aromatic hydrocarbons (PAH) in a single drop of blood.

由于接触致癌物有许多来源和途径，流行病学研究应 使用体内剂量的生物标志物作为致癌物暴露的替代物。然而，直接测量 致癌物质是不切实际的，因为这些化学物质通常是反应性亲电体， 身体作为一种替代方法，致癌物剂量可以通过以下物质的加成产物（加合物）进行研究： 亲电体与血液蛋白，特别是人血清白蛋白（HSA）和血红蛋白（Hb）。因为 这些加合物不会被修复，它们反映了一到两个月内致癌物的剂量。本课题 设想一个“蛋白加合物”，代表给定蛋白质上存在的所有加合物，作为一个实体， 毒理基因组学重要性由于所有亲电子物质都是潜在致癌物质，因此蛋白内收体 与致癌物的发现相比，蛋白质组学和代谢组学更有意义。 尽管它们作为致癌物剂量的生物标志物的效用，有毒化学物质与大量血液的加合物， 蛋白质，如人血清白蛋白（HSA）和Hb，很少用于流行病学研究或 发现人类癌症的始作俑者这主要有三个原因。第一，没有实质性的 加合物富集，HSA和Hb加合物的典型浓度太低，不允许广泛应用 * 发现新的致癌物质。这是“大海捞针的问题”，也困扰着蛋白质组学 调查事务所第二，最先进的质谱法（MS）尚未用于鉴定未知的 蛋白质加合物或分析与人类癌症有关的加合物。第三，很难获得血液 用于测定蛋白质加合物的样品。 在这个项目中，我们假设特定的亚内收体，如HSA的游离半胱氨酸（Cys34）， 可用于发现新的致癌物，并在大型流行病学中作为内照射剂量的生物标志物 问题研究为了考虑这些问题，我们将选择性地从人血中富集HSA的半胱氨酸加合物 然后将使用MS来表征已知和未知的HSA加合物。通过比较加合映射 在淋巴瘤病例和对照组之间，我们将查明可能的致癌物。关于使用 在流行病学研究中，我们将与项目3合作，以确定是否可行 快速测量一滴血中多环芳烃（PAH）的蛋白质加合物。