MOLECULAR DOSIMETRY OF PAH AND ESTROGEN ADDUCTS IN URINE

尿液中多环芳烃和雌激素加合物的分子剂量测定

• 批准号: 6344726
• 负责人: George Pasco Casale
• 金额: $ 16.87万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6344726
• 关键词: DNA damage; adduct; cancer risk; carbopolycyclic compound; chemical carcinogenesis; enzyme linked immunosorbent assay; estrogens; high performance liquid chromatography; immunologic assay /test; laboratory mouse; laboratory rat; monoclonal antibody; technology /technique development; tissue /cell culture; urinalysis

We propose to develop monoclonal antibody-based procedures for molecular dosimetry of depurinating DNA adducts of benzo[alpha]pyrene (BP) and dibenzo[alpha,l]pyrene (DB[alpha,l]P), carcinogens viewed as indicators of exposure to polycyclic aromatic hydrocarbons (PAH) and risk of cancer, and catechol estrogens (CE), putative endogenous procarcinogens. Recent data support an essential contribution of depurinating adducts to cancer initiation by both PAH and CE. Since these adducts are frequently the predominant ones formed, are rapidly lost from the DNA and are excreted in urine, they are potentially useful biomarkers of chemical-specific DNA damage and risk for cancer. This project encompasses the production of monoclonal antibodies (MABs) specific for BP=6=C8Gua(a major depurinating adduct of BP; Aim #1), syn-DB[alpha,l]PDE-14-N7Ade and DB[alpha,l]P-10- N3Ade (major depurinating adducts of DB[alpha,l]P; Aim #2), and 4-OHE1- (1&2)}-N7Gua and 4-OHE2-(1&2)-N7Gua (predominant depurinating adducts of CE; Aim #3). The specific affinities of these MAbs will be determined by competitive enzyme-linked immunosorbent assay (ELISA). In the early stages of development of the proposed molecular dosimeters, MAbs of high specific affinity will be incorporated into competitive ELISAs and immunoaffinity HPLC (1A/HPLC) for quantitation of each adduct added to urine collected from normal rats and humans. Dose-response studies will compare urinary adducts, measured by competitive ELISA or IA/HPLC, with doses of PAH or CE given to rats. Transitional epidemiological studies will apply the MAb- based dosimeters to quantitation of BP adducts in the urine of smokers and nonsmokers, and DB[alpha,l]P adducts in the urine of individuals who are exposed to byproducts of smoky coal combustion and are at high risk for lung cancer. These studies will provide molecular dosimetric procedures needed for prospective epidemiological studies and determination of individual risk for both PAH- and CE-induced cancers.

我们建议开发基于单克隆抗体的分子程序 苯并芘(BP)和脱嘌呤DNA加合物的剂量测定 二苯并[α,l]芘 (DB[α,l]P)，被视为致癌物的指标 接触多环芳烃 (PAH) 和患癌症的风险，以及 儿茶酚雌激素（CE），推定的内源性致癌物。 近期数据 支持脱嘌呤加合物对癌症的重要贡献 由 PAH 和 CE 发起。 由于这些加合物通常是 形成的主要分子，迅速从 DNA 中丢失并排泄到 尿液中，它们是化学特异性 DNA 的潜在有用生物标志物 损害和癌症风险。 该项目包括生产 特异于 BP=6=C8Gua（一种主要的脱嘌呤）的单克隆抗体 (MAB) BP加合物；目标#1)、syn-DB[α,1]PDE-14-N7Ade 和 DB[α,1]P-10- N3Ade（DB[α,l]P 的主要脱嘌呤加合物；目标 #2）和 4-OHE1- (1&2)}-N7Gua 和 4-OHE2-(1&2)-N7Gua（主要脱嘌呤加合物 CE；目标＃3）。 这些 MAb 的具体亲和力将由以下因素确定 竞争性酶联免疫吸附测定（ELISA）。 在早期阶段 所提出的分子剂量计、高特异性单克隆抗体的开发 亲和力将被纳入竞争性 ELISA 和免疫亲和力 HPLC (1A/HPLC) 用于对收集到的尿液中添加的每种加合物进行定量 来自正常老鼠和人类。 剂量反应研究将比较尿 加合物，通过竞争性 ELISA 或 IA/HPLC 与 PAH 或 CE 剂量进行测量 给予老鼠。 过渡流行病学研究将应用 MAb- 基于剂量计对吸烟者尿液中的血压加合物进行定量 非吸烟者和吸烟者尿液中的 DB[α,l]P 加合物 暴露于烟煤燃烧的副产品中，并且面临很高的风险 肺癌。 这些研究将提供分子剂量测定程序 需要前瞻性流行病学研究和确定 PAH 和 CE 诱发癌症的个体风险。