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 花生四烯酸被环氧合酶、脂氧合酶和细胞色素 P450 氧化转化，得到 产生内源性脂质介质，调节体内平衡和疾病的细胞过程。这些脂质 调节剂形成了一个不断发展和扩大的家族，称为类二十烷酸。该应用程序包括两个 两个项目都与类二十烷酸生物化学的新转变有关，并且具有重要意义 使用抑制其生物合成的非甾体抗炎药（NSAID）的后果。这 第一个项目以 5-LOX/COX-2 交叉生物合成途径为中心，而第二个项目是 关注类二十烷酸的新代谢转化，以及这些转化对于血浆的使用意味着什么 和尿前列腺素代谢物作为药物反应的标志物。今天，类二十烷酸的新成员 在脂质组学方法中，经常会发现家族因其与已知类二十烷酸的相似性而被发现。我们有 采用了一种基于理解生物合成途径、酶的结构功能的方法， 和底物使我们能够预测新的转变。这导致了以下发现： 5-LOX/COX-2 交叉生物合成途径形成半缩酮类二十烷酸（HKE2 和 HKD2）和 5- 羟基前列腺素、阿司匹林乙酰化 COX-2 形成的 15R-前列腺素的鉴定以及 鉴定 PGD2 代谢为 11-脱氢- 的 Baeyer-Villiger 氧化途径 血栓素。强调我们方法的相关性的事实是，我们拥有的新型类二十烷酸 在脂质组学分析中尚未鉴定出在体外和体内形成的，可能是由于它们 不寻常的特性使得它们很难在标准分析中检测到。我们设计了分析 允许在体外和体内检测和量化新型类二十烷酸的程序。我们计划继续 鉴定新型类二十烷酸并确定其细胞靶标和体内平衡中的生物学作用 在炎症性疾病模型中。我们正在进行对交叉的生物效应的调查 类二十烷酸已确定受体酪氨酸激酶 (RTK) VEGFR2 作为促血管生成的靶点 HKE2 的活性，以及​​其他 RTK 和介导血小板活化抑制的未知靶标 有待进一步分析。对于 5-羟基前列腺素，我们也计划采用筛选方法 作为前列腺素受体的靶向测试，以确定其细胞靶标并确定生物 影响。我们的目标是鉴定可用于操纵 5-LOX/COX-2 交叉生物合成的化合物。 超过类二十烷酸独立于前列腺素和白三烯的形成。我们将继续 表征前列腺素类化合物的新代谢途径，并建立这些途径在体内的相关性。 新型类二十烷酸的鉴定和表征、其生物学效应以及新型前列腺素代谢 途径将导致对 NSAIDs 可能作用的病理生理状况有更深入的了解 阿司匹林和其他非甾体抗炎药的使用以及对体内前列腺素生物合成的影响。