Novel pathways in eicosanoid biosynthesis and metabolism

类二十烷酸生物合成和代谢的新途径

• 批准号: 10330785
• 负责人: Claus Schneider
• 金额: $ 47.55万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330785
• 关键词: Anabolism; Aspirin; Biochemistry; Biological; Cell physiology; Cytochrome P450; Disease; Eicosanoids; Family; Health; Homeostasis; In Vitro; Inflammation; Inflammatory; Investigation; KDR gene; LOX gene; Leukotrienes; Lipids; Lipoxygenase; Mediating; Metabolic; Metabolic Pathway; Metabolism; Modeling; Non-Steroidal Anti-Inflammatory Agents; PTGS2 gene; Pain; Pathway interactions; Pharmaceutical Preparations; Plasma; Platelet Activation; Procedures; Property; Prostaglandin D2; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Receptor Protein-Tyrosine Kinases; Research; Role; Source; Structure; Testing; Thromboxanes; base; cellular targeting; design; enzyme substrate; hemiketal; in vivo; lipid mediator; lipidomics; member; novel; receptor; response; screening; urinary

Abstract Oxidative transformation of arachidonic acid by cyclooxygenases, lipoxygenases, and cytochromes P450 gives rise to endogenous lipid mediators that regulate cellular processes in homeostasis and disease. These lipid mediators form an evolving and expanding family referred to as eicosanoids. This application comprises two projects that both are concerned with novel transformations in eicosanoid biochemistry and present important ramifications for the use of non-steroidal anti-inflammatory drugs (NSAIDs) that inhibit their biosynthesis. The first project is centered around the 5-LOX/COX-2 cross-over biosynthetic pathway while the second project is concerned with novel metabolic transformations of eicosanoids, and what these mean for the use of plasma and urinary prostanoid metabolites as markers of drug response. Today, novel members of the eicosanoid family are often discovered in lipidomics approaches by their similarity with known eicosanoids. We have employed an approach based on understanding the structure-function of the biosynthetic pathways, enzymes, and substrates that allowed us to make predictions of novel transformations. This has led to the discovery of the 5-LOX/COX-2 cross-over biosynthetic pathway forming hemiketal eicosanoids (HKE2 and HKD2) and 5- hydroxy-prostaglandins, the identification of 15R-prostaglandins formed by aspirin-acetylated COX-2, and the identification of the Baeyer-Villiger oxidative pathway underlying the metabolism of PGD2 to 11-dehydro- thromboxanes. Underscoring the relevance of our approach is the fact that the novel eicosanoids we have shown to be formed in vitro and in vivo have not been identified in lipidomics analyses, likely due to their unusual properties that make them difficult to detect in standard analyses. We have designed analytical procedures that allow to detect and quantify the novel eicosanoids in vitro and in vivo. We plan to continue the identification of novel eicosanoids and to establish their cellular targets and biological role in homeostasis and in models of inflammatory disease. Our ongoing investigation into the biological effects of the cross-over eicosanoids has identified the receptor tyrosine kinase (RTK) VEGFR2 as a target for the pro-angiogenic activity of HKE2, as well as other RTK and an unknown target that mediates inhibition of platelet activation that are to be further analyzed. For the 5-hydroxy-prostaglandins we plan to employ screening approaches as well as targeted testing of prostanoid receptors in order to identify their cellular targets and determine biological effects. We aim to identify compounds that can be used to manipulate biosynthesis of 5-LOX/COX-2 cross- over eicosanoids independent from the formation of prostaglandins and leukotrienes. We will continue to characterize novel metabolic pathways of prostanoids and establish the relevance of these pathways in vivo. Identification and characterization of novel eicosanoids, their biological effects, and novel prostanoid metabolic pathways will result in a refined understanding of the pathophysiologic conditions for which NSAIDs may be used and effect of aspirin and other NSAIDs on prostanoid biosynthesis in vivo.

抽象的 通过环氧酶，脂氧酶和细胞色素P450氧化转化的蛛网膜酸会产生 在调节体内平衡和疾病中细胞过程的内源性脂质介质的上升。这些脂质 介体形成了一个不断发展的家族，称为类eicosanoids。该应用程序包括两个 两者都关注eicosanoid生物化学的新型转变并提出重要的项目 使用抑制其生物合成的非甾体类抗炎药（NSAIDS）的影响。这 第一个项目以5-lox/cox-2交叉生物合成途径为中心，第二个项目是 与eicosanoids的新代谢转化有关，以及这些对血浆的使用意味着什么 和前列腺素代谢产物作为药物反应的标志物。今天，eicosanoid的新颖成员 在脂肪组学方法中，通常通过与已知类花生酸的相似性发现了家庭。我们有 采用了一种基于理解生物合成途径的结构功​​能的方法，酶， 以及使我们能够预测新型转换的底物。这导致发现 5-lox/cox-2交叉生物合成途径，形成半蛋白酶（HKE2和HKD2）和5-- 羟基促素蛋白，鉴定由阿司匹林乙酰化COX-2形成的15R-局丁氏蛋白，并鉴定 鉴定Baeyer-Villiger氧化途径是PGD2至11-脱氧的代谢的基础 血栓盒。强调我们方法的相关性是我们拥有的新颖的eicosanoids 在脂质组学分析中尚未确定在体外和体内形成的 在标准分析中难以检测的异常属性。我们设计了分析 可以在体外和体内检测和量化新颖的类固醇的程序。我们计划继续 鉴定新型eicosanoids，并在体内平衡和生物学中建立其细胞靶标的 在炎症性疾病模型中。我们正在进行对交叉生物学作用的研究 类花生酸已经确定受体酪氨酸激酶（RTK）VEGFR2是促血管生成的靶标 HKE2的活性以及其他RTK和一个未知的靶标，介导了抑制血小板激活的抑制 要进一步分析。对于5-羟基 - 前蛋白，我们还计划采用筛选方法 作为前列腺素受体的靶向测试，以鉴定其细胞靶标并确定生物学 效果。我们旨在识别可用于操纵5-lox/cox-2横界生物合成的化合物 在类素烷的上，独立于前列腺素和白细胞的形成。我们将继续 表征前列腺素的新代谢途径，并在体内建立这些途径的相关性。 新颖的类类，其生物学作用和新的前列腺素代谢的鉴定和表征 途径将导致对NSAID可能为病理生理状况的精致理解 阿司匹林和其他NSAID对体内前列腺素生物合成的影响和影响。