MOLECULAR DOSIMETRY OF PAH AND ESTROGEN ADDUCTS IN URINE

我们建议开发单克隆抗体为基础的程序的分子 苯并[α]芘（BP）和 二苯并[alpha，l]芘（DB[alpha，l]P），致癌物质被视为 暴露于多环芳烃（PAH）和癌症风险，以及 儿茶酚雌激素（CE），推定的内源性前致癌物。 最近的数据 支持脱嘌呤加合物对癌症的重要贡献 由PAH和CE启动。 由于这些加合物通常是 主要的形成，迅速从DNA中丢失，并排出体外。 尿液中，它们是潜在有用的化学特异性DNA的生物标志物 损害和癌症风险。 该项目包括生产 对BP=6= C8 Gua（一种主要的脱嘌呤酶）具有特异性的单克隆抗体（MAB） BP的加合物;目标#1）、顺式-DB [α，1]PDE-14-N7 Ade和DB[α，1]P-10-N7 Ade。 N3 Ade（DB[α，1]P的主要脱嘌呤加合物;目标#2）和4-OHE 1- （1&2）}-N7 Gua和4-OHE 2-（1&2）-N7 Gua（主要的脱嘌呤加合物 CE;目标#3）。 这些MAb的特异性亲和力将通过以下测定： 竞争性酶联免疫吸附测定（ELISA）。 在早期阶段 发展的建议分子剂量计，单克隆抗体的高特异性 亲和性将被纳入竞争性ELISA和免疫亲和性 HPLC（1A/HPLC）用于定量采集的尿液中添加的各加合物 从正常的老鼠和人类。 剂量反应研究将比较尿液 加合物，通过竞争性ELISA或IA/HPLC测定，PAH或CE剂量 给老鼠。 过渡性流行病学研究将使用MAb- 基于剂量计定量吸烟者尿液中的BP加合物， 不吸烟者，和DB[α，l]P加合物的个人谁是尿 暴露于冒烟的煤炭燃烧的副产品，并处于高风险之中， 肺癌 这些研究将提供分子剂量测定程序 进行前瞻性流行病学研究和确定 PAH和CE诱导的癌症的个体风险。

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.