LC-MS Analysis of DNA Adducts of Aromatic Amines: Relationship to Bladder Cancer

芳香胺 DNA 加合物的 LC-MS 分析：与膀胱癌的关系

• 批准号: 8513263
• 负责人: Paul Vouros
• 金额: $ 21.03万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-18 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8513263
• 关键词: 4-biphenylamine; Address; Affect; Aromatic Amines; Base Sequence; Binding; Biological; Biological Availability; Biological Markers; Bladder; Buffaloes; Carcinogens; Cell Line; Cells; Chemicals; Clinical; Clinical Research; Collaborations; Computer software; Coupling; DNA; DNA Adduction; DNA Adducts; DNA Damage; Data; Detection; Development; Digestion; Effectiveness; Environment; Etiology; Evaluation; Event; Exposure to; Food; Funding; Genes; Genetic; Goals; Grant; Hair Dyes; Health; High Pressure Liquid Chromatography; Hot Spot; Human; Human Volunteers; In Vitro; Investigation; Life; Link; Liquid Chromatography; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mass Spectrum Analysis; Measurement; Metabolic Activation; Methodology; Methods; Minor; Molecular; Mutagens; Oligonucleotides; Organ; Phase; Physiological; Positioning Attribute; Procedures; Process; Progress Reports; Protocols documentation; Research; Research Design; Risk; Role; Roswell Park Cancer Institute; Sampling; Site; Smoker; Source; Spectrometry; TP53 gene; Techniques; Technology; Testing; Tissues; Urine; Urothelial Cell; Work; Xenobiotics; adduct; base; carcinogenesis; cigarette smoking; cigarette smoking; clinical application; environmental chemical; exposed human population; genotoxicity; improved; in vivo; inhibitor/antagonist; innovation; ion mobility; meetings; nano-electrospray; new technology; novel; prevent; programs; smoking cessation; success; tool

Summary Statement The covalent binding of genotoxins to DNA to form DNA adducts is considered to be a first step in chemically induced carcinogenesis. Measurement of DNA adducts in cells and tissues provides direct evidence of genetic damage and a means for assessing human health risk from exposure. However, the low levels at which adducts occur in affected organs and the limited amounts of DNA frequently available make their accurate measurement a highly challenging task. Methodology based on microcapillary liquid chromatography in combination with nano-electrospray mass spectrometry developed in this program has enabled us to achieve detection and quantification capabilities of adducts at levels similar to those frequently encountered in human exposure using low microgram quantities of DNA. Aromatic amines, compounds found in cigarette smoke, hair dyes and other environmental sources have been implicated in a variety of cancers including bladder cancer. 4-Aminobiphenyl (4-ABP), is a major carcinogen in this class of compounds and recent evidence has identified a potential relationship between mutational hot spots of the p53 gene in human bladder cancer and adduction sites of 4-ABP. A highly sensitive analytical protocol will be developed to quantify the DNA adducts of 4-ABP in urothelial cells from human volunteers involved in a smoke / cessation clinical study in order to assess the link between cigarette smoking and the risk for carcinogenesis. Use of urine as the physiological medium provides a non-invasive procedure which is more amenable for incorporation into human studies to assess the risk for bladder cancer from environmental chemicals. Agents which inhibit the formation of aromatic amine-DNA adducts will be tested in a human bladder cell line, thereby providing a broader perspective of their biological significance in their induction of bladder cancer. Development of methodology to characterize oligonucleotide adducts will expand the breadth of these clinical studies and will allow us to gain a better understanding of adduct formation in the context of base sequence and potentially identify the adduction in terms of specific domains of DNA and selected gene sequences. Additional novel technologies proposed here based on differential ion mobility spectrometry - mass spectrometry (DMS - MS) and the coupling of HPLC with MS and microNMR, will introduce new powerful tools to facilitate and improve the state of the art in the analysis of DNA adducts. We expect that the results of this work will help establish more definitively the degree of involvement of DNA adducts in the etiology of human cancer and define the relationship between bladder cancer and cigarette smoke.

简要说明 遗传毒素与DNA共价结合形成DNA加合物被认为是遗传毒性的第一步。 化学致癌作用细胞和组织中DNA加合物的测量提供了直接的 遗传损害的证据和评估暴露对人类健康风险的手段。但 在受影响的器官中发生的加合物水平较低，并且经常可获得的DNA量有限 使其精确测量成为一项极具挑战性的任务。基于微毛细管液体的方法学 本项目开发的纳米电喷雾质谱联用色谱法 使我们能够实现加合物的检测和定量能力， 在人类接触低微克量DNA时经常遇到。 芳香胺，在香烟烟雾中发现的化合物，染发剂和其他环境来源， 与包括膀胱癌在内的多种癌症有关。4-氨基联苯（4-ABP）是一种主要的 致癌物质在这类化合物和最近的证据已经确定了潜在的关系 人膀胱癌p53基因突变热点与4-ABP加合位点之间的关系。一 将开发一种高灵敏度的分析方案来定量尿路上皮中4-ABP的DNA加合物 来自参与吸烟/戒烟临床研究的人类志愿者的细胞， 吸烟和致癌风险之间的联系。使用尿液作为生理介质 提供了一种非侵入性的方法，其更适合于结合到人类研究中， 评估环境化学物质对膀胱癌的风险。抑制以下物质形成的药剂 芳香胺-DNA加合物将在人膀胱细胞系中进行测试，从而提供更广泛的 展望其在诱导膀胱癌中的生物学意义。发展 表征寡核苷酸加合物的方法将扩大这些临床研究的广度， 将使我们能够更好地理解在碱基序列的背景下加合物的形成， 潜在地根据DNA的特定结构域和选定的基因序列来鉴定加合。 本文提出的基于差分离子迁移谱-质谱的其他新技术 质谱（DMS-MS）和HPLC与MS和microNMR的耦合，将引入新的 强大的工具，以促进和改善最先进的DNA加合物的分析。我们预计 这项工作的结果将有助于更明确地建立DNA加合物参与的程度， 人类癌症的病因学，并确定膀胱癌和吸烟之间的关系。

项目成果

Microscale epitope mapping by affinity capillary electrophoresis-mass spectrometry.

• DOI: 10.1021/ac9700944
• 发表时间: 1997-08
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Y. Lyubarskaya;Y. Dunayevskiy;P. Vouros;B. Karger

Interfacing of CE in a PVP matrix to ion trap mass spectrometry: analysis of isomeric and structurally related (N-acetylamino)fluorene-modified oligonucleotides.

PVP 基质中 CE 与离子阱质谱的接口：异构体和结构相关的（N-乙酰氨基）芴修饰寡核苷酸的分析。

• DOI: 10.1021/ac9713823
• 发表时间: 1998
• 期刊: Analytical chemistry
• 影响因子: 7.4
• 作者: Harsch,A;Vouros,P
• 通讯作者: Vouros,P

Accurate and rapid modeling of iron-bleomycin-induced DNA damage using tethered duplex oligonucleotides and electrospray ionization ion trap mass spectrometric analysis.

使用系链双链寡核苷酸和电喷雾电离离子阱质谱分析对铁博来霉素诱导的 DNA 损伤进行准确、快速的建模。

• DOI: 10.1093/nar/28.9.1978
• 发表时间: 2000
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Harsch,A;Marzilli,LA;Bunt,RC;Stubbe,J;Vouros,P
• 通讯作者: Vouros,P

Formation and analysis of heterocyclic aromatic amine-DNA adducts in vitro and in vivo.

体外和体内杂环芳香胺-DNA 加合物的形成和分析。

• DOI: 10.1016/j.jchromb.2003.10.053
• 发表时间: 2004
• 期刊: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.
• 作者: Turesky,RobertJ;Vouros,Paul
• 通讯作者: Vouros,Paul

Metabolic stability of 3-epi-1α,25-dihydroxyvitamin D3 over 1 α 25-dihydroxyvitamin D3: metabolism and molecular docking studies using rat CYP24A1.

3-epi-1α,25-二羟基维生素 D3 超过 1 α 25-二羟基维生素 D3 的代谢稳定性：使用大鼠 CYP24A1 进行代谢和分子对接研究。

• DOI: 10.1002/jcb.24576
• 发表时间: 2013
• 期刊: Journal of cellular biochemistry
• 影响因子: 4
• 作者: Rhieu,SteveY;Annalora,AndrewJ;Wang,Guochun;Flarakos,CarolineC;Gathungu,RoseM;Vouros,Paul;Sigüeiro,Rita;Mouriño,Antonio;Schuster,Inge;Palmore,GTayhasR;Reddy,GSatyanarayana
• 通讯作者: Reddy,GSatyanarayana