Prevention of COX-2 Derived DNA and Histone Modifications in Cancer

预防癌症中 COX-2 衍生的 DNA 和组蛋白修饰

基本信息

批准号：
9248194
负责人：
JOHN Alexander OATES
金额：
$ 15.85万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9248194
关键词：
Accounting Address Amines Animal Model Biology Breast Cancer cell line Carcinogens Cell Nucleus Cells Cessation of life Chromosomal Instability Colon Colon Carcinoma Controlled Clinical Trials Cytidine DNA DNA Adducts DNA Methylation DNA Modification Process Deoxycytidine Development Dinoprostone Effectiveness Enzymes Epidemiologic Studies Epigenetic Process Epithelial Cells Event Evolution Genes Genomics Growth HMGA1 gene Health High Pressure Liquid Chromatography Histones Histopathology Human Intervention Investigation Lactams Lipid Peroxidation Lung Lysine Malignant - descriptor Malignant Neoplasms Malignant neoplasm of lung Malondialdehyde Metabolic Biotransformation Methods Modification Mus Mutagenesis Neoplasms Pancreas Prevention Prostaglandin D2 Prostaglandin-Endoperoxide Synthase Prostaglandins Protein Isoforms Proteins Pyrroles Research Sampling Series Structure Testing Thromboxane A2 Time Transcription Alteration Transgenic Mice Tumor Suppressor Genes Uterine Corpus Sarcoma Xenograft procedure adduct base cancer prevention carcinogenesis cyclooxygenase 2 histone modification in vivo inhibitor/antagonist insight ketoaldehyde malignant phenotype new technology novel novel strategies prevent stable isotope tool tumor tumorigenesis

Accounting Address Amines Animal Model Biology Breast Cancer cell line Carcinogens Cell Nucleus Cells Cessation of life Chromosomal Instability Colon Colon Carcinoma Controlled Clinical Trials Cytidine DNA DNA Adducts DNA Methylation DNA Modification Process Deoxycytidine Development Dinoprostone Effectiveness Enzymes Epidemiologic Studies Epigenetic Process Epithelial Cells Event Evolution Genes Genomics Growth HMGA1 gene Health High Pressure Liquid Chromatography Histones Histopathology Human Intervention Investigation Lactams Lipid Peroxidation Lung Lysine Malignant - descriptor Malignant Neoplasms Malignant neoplasm of lung Malondialdehyde Metabolic Biotransformation Methods Modification Mus Mutagenesis Neoplasms Pancreas Prevention Prostaglandin D2 Prostaglandin-Endoperoxide Synthase Prostaglandins Protein Isoforms Proteins Pyrroles Research Sampling Series Structure Testing Thromboxane A2 Time Transcription Alteration Transgenic Mice Tumor Suppressor Genes Uterine Corpus Sarcoma Xenograft procedure adduct base cancer prevention carcinogenesis cyclooxygenase 2 histone modification in vivo inhibitor/antagonist insight ketoaldehyde malignant phenotype new technology novel novel strategies prevent stable isotope tool tumor tumorigenesis

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项目摘要

DESCRIPTION (provided by applicant): The hypothesis that highly reactive metabolites of the cyclooxygenase (COX) enzymes contribute to the COX-dependent growth of cancers will be examined in the proposed research. A concerted body of evidence indicates that the COXs contribute to the evolution of malignant neoplasms, and it includes findings from epidemiological studies and controlled clinical trials as well as investigations in animal models and cells. The inducible COX-2 isoform is expressed in many epithelial cell tumors, including those of the colon, lung, breast, and pancreas. The product of the COXs is prostaglandin H2 (PGH2) which undergoes cell specific enzymatic biotransformation to the prostaglandins D2, E2, F2a, I2, and thromboxane A2. All current research on the mechanism by which the COXs contribute to cancer addresses the actions of these enzymatically derived prostanoids. However, PGH2 also undergoes non-enzymatic rearrangement yielding the highly reactive -ketoaldehydes, levuglandins (LG) E2 and D2 as 20% of the total products. The LGs react almost instantaneously to form covalent adducts of primary amines. We have demonstrated that LGE2 forms a covalent adduct with deoxycytidine bases in DNA. The stable form of the adduct has been characterized as a pyrrole, and we have developed an HPLC/tandem mass spectrometric analysis for it, employing stable isotope dilution. This has permitted identification of COX-2 dependent formation of the LG-DNA adduct in two cancer cell lines. The formation of this bulky COX-derived covalent adduct suggests that it could engender mutagenesis and/or chromosomal instability. In addition to this modification of DNA, LG reacts with the epsilon-amine of lysine to form stable adducts with a lactam structure. COX-2 derived LG-histone adducts also have been identified in a human lung cancer cell line, raising the possibility of epigenetic alteration of the transcription of driver genes or tumor suppressor genes. We have developed a potent scavenger of LGs, ethyl-salicylamine, that prevents formation of LG adducts of DNA and histones but does not inhibit the COXs. It also scavenges other reactive carbonyls such as malondialdehyde which is a product of the cyclooxygenases and of lipid peroxidation. The proposed research will employ ethyl-salicylamine as a tool with which to address the hypothesis that modification of DNA and histone by reactive carbonyls such as LG and malondialdehyde contribute to the growth of COX-2 dependent cancers. The presence of these DNA and histone modifications in in vivo cancers as well as reduction of these modifications by ethyl-salicylamine will be determined to verify the efficacy of the pharmacologic intervention. COX-2 is associated with highly malignant cancers that result in >200,000 deaths annually in the US alone. Accordingly, understanding how it contributes to the evolution of malignancy is important.

描述（由适用提供）：在拟议的研究中将检查环氧合酶（COX）酶的高反应性代谢物（COX）酶的高度反应性代谢产物的假设。一致的证据表明，考克斯有助于恶性肿瘤的进化，其中包括流行病学研究和对照临床试验的发现以及对动物模型和细胞的投资。诱导型COX-2同工型在许多上皮细胞肿瘤中表达，包括结肠，肺，乳腺癌和胰腺的肿瘤。 Coxs的乘积是前列腺素H2（PGH2），它经过细胞特异性的酶促生物转化对前列腺素D2，E2，F2A，I2和Thromboxane A2。目前，关于COX促进癌症的机制的所有研究均解决了这些酶促衍生的前列腺素的作用。然而，PGH2还经历非酶排列，产生高反应性的ket酮醛，左guglandins（LG）E2和D2，占总产物的20％。 LGS几乎瞬间反应形成了初级胺的共价加合物。我们已经证明LGE2在DNA中与脱氧胞苷碱形成共价加合物。加合物的稳定形式已被描述为吡咯，我们采用了稳定的同位素稀释度开发了HPLC/串联质谱分析。这允许鉴定在两个癌细胞系中LG-DNA加合物的COX-2依赖性形成。这种庞大的Cox衍生的共价加合物的形成表明它可能导致诱变和/或染色体不稳定性。除了这种DNA的修饰外，LG还与赖氨酸的伊氏胺反应，形成具有乳糖结构的稳定加合物。在人肺癌细胞系中也发现了COX-2衍生的LG-histone加合物，从而增加了驱动基因或肿瘤抑制基因转录的表观遗传改变的可能性。我们已经开发了潜在的LGS乙基水杨胺的清除剂，可防止形成DNA和组蛋白的LG加合物，但不抑制COX。它还清除了其他反应性羰基，例如丙二醛，它是环氧酶和脂质过氧化的产物。拟议的研究将利用乙基酰胺作为一种工具来解决以下假设：通过反应性羰基（如LG和丙二醛）对DNA和组蛋白的修饰有助于COX-2依赖性癌症的生长。这些DNA和组蛋白在体内癌症中的存在以及通过乙基甲基胺对这些修饰的减少将确定以验证药物干预的效率。 COX-2与高度恶性肿瘤有关，仅在美国，每年就会导致> 200,000人死亡。根据了解，它如何促进恶性肿瘤的演变很重要。