MOLECULAR DOSIMETRY OF PAH AND ESTROGEN ADDUCTS IN URINE

我们建议开发基于单抗的分子检测程序。 苯并[α]芘(BP)和脱氧核糖核酸(DNA)加合物的剂量学研究 二苯并[α，L]芘(Db[α，L]P)，被视为 暴露于多环芳烃(PAH)与癌症风险，以及 儿茶酚雌激素(CE)，推测为内源性致癌物质。最新数据 支持净化加合物对癌症的重要贡献 由多环芳烃和CE共同引发。因为这些加合物通常是 形成的主要DNA，迅速从DNA中丢失，并在体内排泄 尿液，它们可能是化学特异性DNA的有用生物标记物 损害和患癌症的风险。这个项目包括生产 抗BP=6=C8Gua(一种主要的大便)的单抗(MAB) BP的加合物；目的#1)，syn-DB[α，L]PDE-14-N7Ade和DB[α，L]P-10- N3Ade(DB[α，L]P；Aim#2)的主要精制加合物，和4-OHE1- (1和2)}-N7Gua和4-OHE2-(1和2)-N7Gua(主要的净化加合物 目标3)。这些单抗的具体亲和力将由以下因素确定 竞争性酶联免疫吸附试验(EL ISA)。在早期阶段 发展建议的分子剂量计，高比特效的单抗 亲和力将被纳入竞争性ELISA和免疫亲和力 用于定量添加到所收集的尿液中的每个加合物的高效液相色谱(1A/HPLC) 从正常的老鼠和人类身上。剂量-反应研究将比较尿液 加合物，用竞争性酶联免疫吸附试验或IA/HPLC法测定，含有PAH或CE剂量 给老鼠吃的。过渡性流行病学研究将应用MAb- 基于剂量计的吸烟者尿液中BP加合物的定量 非吸烟者，和尿液中的DB[α，L]P加合物 暴露在烟雾弥漫的煤燃烧副产品中，并有很高的患病风险 肺癌。这些研究将提供分子剂量学程序。 进行前瞻性流行病学研究和确定 PAH和CE诱发癌症的个体风险。

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.