Molecular dosimetry of endogeneous genotoxins

内源性基因毒素的分子剂量测定

基本信息

批准号：
7893041
负责人：
Ian Alexander Blair
金额：
$ 24.88万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-05-18 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7893041
关键词：
Acids Address Animal Model Animal Tarsus Arachidonate 15-Lipoxygenase Arachidonate 5-Lipoxygenase Arachidonic Acids Ascorbic Acid Atmospheric Pressure Award Biological Markers Calendar Cancer Etiology Cancer Model Carbon Cell model Cells Chemicals Colon Colon Carcinoma Colorectal Cancer Complement Cytidine DNA DNA Adducts DNA Damage DNA Repair Enzymes DNA Sequence Rearrangement Deoxycytidine Deoxyguanosine Discipline Dose Eicosanoids Eicosatetraenoic Acids Electrons Electrospray Ionization Endothelial Cells Epithelial Cells Extracellular Fluid Family Glutathione Glutathione S-Transferase Human Hydrogen Peroxide In Vitro Inbred C57BL Mice Intestines Isoprostanes Lipid Peroxidation Lipid Peroxides Lipids Liquid Chromatography Malignant Neoplasms Manuscripts Mass Spectrum Analysis Measurement Mediating Metabolism Methodology Modeling Molecular Monitor Mus Mutagens N-methylacetamide-oxotremorine M Normal tissue morphology Oxidative Stress Pathway interactions Phase Play Polyunsaturated Fatty Acids Prize Production Prostaglandin-Endoperoxide Synthase Proteins RNA Rattus Reaction Reactive Oxygen Species Relative (related person)Reporting Research Research Personnel Risk Role Sensitivity and Specificity Site Structure System Techniques Testing Time Tissues Urine adduct analog carboxylate cell type cyclooxygenase 2 dosimetry enantiomer in vivo in vivo Model innovation insight instrumentation ionization member mouse model multiple reaction monitoring novel octadecadienoic acid programs stable isotope tandem mass spectrometry

项目摘要

DESCRIPTION (provided by applicant): The analysis of DNA-adducts is important for molecular dosimetry studies, and for monitoring treatment options because they can provide insight into the amount of genotoxin that that has reached the DNA in the tissue under study. No specific biomarkers of endogenous lipid hydroperoxide-mediated DNA damage in vivo are available in spite of an intensive research effort spanning several decades. Our studies over the last six years have identified three heptanone-etheno-DNA-adducts that can only arise from endogenously formed lipid hydroperoxides. We have now shown that heptanone-etheno-2'-deoxyguanosine heptanone-etheno-21- deoxy-cytidine adducts are formed in the DNA of rat intestinal epithelial cells that stably express COX-2 and that the adducts are present in mammalian tissue DNA. We have also discovered a new class of cyclic glutathione (GSH)-adducts derived from endogenous lipid hydroperoxides. Lipid hydroperoxides undergo homolytic decomposition to the highly reactive bifunctional electrophile 4-oxo-2(E)-nonenal (ONE). Lipid hydroperoxide-derived ONE is responsible for the formation of both heptanone-etheno- adducts and the novel cyclic GSH-adducts. Our recent discovery that electron capture atmospheric chemical ionization mass spectrometry can be used to analyze chiral lipids with high sensitivity and specificity makes it possible to analyze the lipid hydroperoxide precursors of DNA- and glutathione- adducts. Therefore, we are poised to make some significant advances in understanding the role of oxidative stress in the etiology of cancer. We propose to address the following hypotheses: 1. Endogenous GSH-adducts and their metabolites represent a new class of biomarkers that will complement isoprostanes as biomarkers of oxidative stress. 2. Endogenous DNA-, RNA-, and cyclic GSH-adducts adducts can arise from cyclooxygenase-mediated pathways. 3. Endogenous DNA-, RNA-, and cyclic GSH-adducts can arise from 5-lipoxygenase but not 15-lipoxygenase. 4. Heptanone- etheno-DNA- and RNA-adducts, together with metabolites of endogenous cyclic GSH-adducts are dosimeters of colon cancer. The proposed research will be conducted under four specific aims. Aim 1. To examine the enzymatic formation and metabolism of endogenous lipid hydroperoxide-derived GSH- adducts. Aim 2. To compare ROS- and COX-2-mediated formation of endogenous DNA-, RNA-, and GSH-adducts with lipid hydroperoxide formation in cellular models. Aim 3. To compare ROS- and LOX- mediated formation of endogenous DNA-, RNA-, and GSH-adducts with lipid hydroperoxide formation in cellular models. Aim 4. To quantify endogenous lipid hydroperoxide-derived DNA-adducts, RNA- adducts, GSH-adducts as potential cancer biomarkers in animal models of colon cancer and in human colon tissue.

描述（由申请人提供）：DNA添加剂的分析对于分子剂量学研究和监测治疗方案很重要，因为它们可以深入了解已研究组织中DNA的基因毒素量。尽管经过数十年的大量研究工作，但在体内尚无内源性脂质氢过氧介导的DNA损伤的特定生物标志物。在过去的六年中，我们的研究确定了只有内生形成的脂质氢过氧化物才能引起的三个己酮 - 乙烯-DNA-DNA添加剂。现在，我们已经表明，在大鼠肠上皮细胞的DNA中形成了己酮-2'-脱氧鸟酮-21-脱氧 - 酪蛋白加合物，稳定表达cox-2，并且在哺乳动物组织DNA中存在加合物。我们还发现了一类新的循环谷胱甘肽（GSH） - 源自内源性脂质氢过氧化物。脂质氢过氧化物对高反应性双功能电力4-oxo-2（e） - 硝酸（一个）经历均质分解。脂质氢过氧化物衍生的人是甲酮 - 乙烯加合物和新型环状GSH添加剂的形成。我们最近发现，电子捕获大气化学电离质谱法可用于分析具有高灵敏度和特异性的手性脂质，因此可以分析DNA和谷胱甘肽加合物的脂质氢过氧前体。因此，我们准备在理解氧化应激在癌症病因中的作用方面取得重大进步。我们建议解决以下假设：1。内源性GSH-添加剂及其代谢物代表了一种新的生物标志物，它们将与异丙烷相辅相成，作为氧化应激的生物标志物。 2。内源性DNA，RNA和环状GSH添加物可以来自环氧酶介导的途径。 3。内源性DNA-，RNA-和环状GSH-添加剂可能来自5-脂氧酶，而不是15-脂氧酶。 4。己酮 - 埃乙烯-DNA和RNA-ADDUCTS，以及内源环状GSH添加剂的代谢产物，是结肠癌的剂量计。拟议的研究将以四个特定目的进行。目的1。检查内源性脂质氢过氧化物加合物的酶形成和代谢。目的2。在细胞模型中，将内源性DNA，RNA和GSH添加剂的ROS-和COX-2介导的内源性DNA，RNA和GSH添加剂与脂质氢过氧化物形成进行比较。目的3。在细胞模型中，将内源性DNA，RNA和GSH添加剂的ROS-和LOX介导的形成与脂质氢过氧化物形成。目的4。量化内源性脂质氢过氧化物衍生的DNA-添加剂，RNA加合物，GSH添加剂，作为结肠癌动物模型和人类结肠组织中的潜在癌症生物标志物。

项目成果

期刊论文数量（35）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

5-Lipoxygenase-mediated endogenous DNA damage.

DOI：
10.1074/jbc.m109.011841
发表时间：
2009-06-19
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Jian W;Lee SH;Williams MV;Blair IA
通讯作者：
Blair IA

Targeted chiral lipidomics analysis of bioactive eicosanoid lipids in cellular systems.

DOI：
10.5483/bmbrep.2009.42.7.401
发表时间：
2009-07-31
期刊：
BMB reports
影响因子：
3.8
作者：
Lee SH;Blair IA
通讯作者：
Blair IA

20-HETE mediates ozone-induced, neutrophil-independent airway hyper-responsiveness in mice.

DOI：
10.1371/journal.pone.0010235
发表时间：
2010-04-20
期刊：
PloS one
影响因子：
3.7
作者：
Cooper PR;Mesaros AC;Zhang J;Christmas P;Stark CM;Douaidy K;Mittelman MA;Soberman RJ;Blair IA;Panettieri RA
通讯作者：
Panettieri RA

Analysis of endogenous glutathione-adducts and their metabolites.