Novel Lipid 2nd Messengers Regulating Bioenergetics and Signaling in Human Myocardium

调节人体心肌生物能和信号传导的新型脂质第二信使

• 批准号: 10593961
• 负责人: RICHARD W GROSS
• 金额: $ 58.12万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10593961
• 关键词: 12-HETE; Ablation; Address; Arachidonate 12-Lipoxygenase; Arachidonate 15-Lipoxygenase; Attenuated; Binding; Bioenergetics; CRISPR/Cas technology; Cardiac; Cardiac Myocytes; Cardiolipins; Cardiovascular Diseases; Cardiovascular system; Carnitine; Cause of Death; Cell Death; Cells; Chemicals; Chemistry; Citric Acid Cycle; Complex; Congestive Heart Failure; Derivation procedure; Developed Countries; Eicosanoids; Endothelial Cells; Environment; Enzymes; Female; Fibroblasts; Free Radicals; Functional disorder; Future; G-Protein-Coupled Receptors; Generations; Glutathione Disulfide; Grant; Heart; Heart Diseases; Heart Mitochondria; Heart failure; Human; Hydrolysis; Hydroxyeicosatetraenoic Acids; Immune system; Incubated; Industrialization; Inflammation; Inflammation Mediators; Inflammatory; Iron; Iron Chelation; Ketone Bodies; Knowledge; Label; Ligands; Lipid Peroxides; Lipids; Lysophosphatidylcholines; Lysophospholipids; Macrophage; Mediating; Membrane; Metabolic; Metabolic Pathway; Metabolism; Mitochondria; Molecular; Myocardial Ischemia; Myocardium; NADP; Oxidases; Oxidation-Reduction; Oxidative Stress; Palmitates; Parents; Pathway interactions; Phospholipases A; Phospholipids; Plasmalogens; Play; Predisposition; Production; Proteins; Publishing; Pyruvate; Reaction; Reactive Oxygen Species; Research; Resolution; Respiration; Role; Signal Transduction; Societies; Specificity; Stress; Substrate Cycling; Succinates; Technology; Ubiquinone; beta-Hydroxybutyrate; cell type; cellular targeting; chemoproteomics; cyclooxygenase 2; cytochrome c; cytokine; diagnostic signature; experimental study; inhibitor; insight; interest; lipid mediator; lipid metabolism; lipidome; male; monocyte; novel; oxidation; oxidized lipid; polyunsaturated fat; protein complex; receptor; research study; response; stable isotope; targeted treatment; ubiquinol

ABSTRACT Cardiovascular disease is the most common cause of death in industrialized nations. During the course of our studies, we have identified previously undiscovered mitochondrial pathways of lipid metabolism and signaling which lead to the generation of 2-arachidonoyl-lysophosphatidylcholine (2-AA-LPC) and 2-arachidonoyl- lysophosphatidylethanolamine (2-AA-LPE). These include: 1) the identification of iPLA2g (PNPLA8) as a phospholipase with sn-1 specificity; and 2) oxidized cardiolipin-activated cytochrome c serving as a plasmalogenase catalyzing the production of 2-AA LPC and 2-AA LPE. Further research demonstrated that 2-AA-LPC and 2-AA-LPE are excellent substrates for cyclooxygenase-2 resulting in a plethora of unanticipated metabolites. Remarkably, incubation of these substrates with either 12-lipoxygenase or 15-lipoxygenase resulted in their oxidation to 12- H(p)ETE- or 15-H(p)ETE-lysophospholipids. Building upon these discoveries we identified 2-AA-LPC and 2-AA-LPE as signaling and metabolic nodes in lipid synthesis and human heart mitochondrial function. Importantly, we have identified the ability of failing human heart mitochondria to generate increased amounts of HETE eicosanoids in response to Ca2+ challenge in comparison to non-failing control mitochondria. Recently, ferroptosis has been identified as a mechanism that leads to cell death through the accumulation of lipid hydroperoxides. During myocardial ischemia and heart failure, a substantial portion of mitochondrial iron (Fe+3) is released from its bound state to become free Fe+2 that initiates the formation of reactive oxygen species (ROS) through Haber-Weiss and Fenton-type chemistries. The present research is targeted to identifying the roles of prominent mechanisms responsible for the oxidized lipids and their roles in membrane dysfunction in failing myocardium. The proposed research will focus on mechanistically understanding the mechanisms leading to oxidized lipid production in the failing human heart. Specifically, we will identify the chiral enrichment in different classes of oxidized lipids to gain mechanistic insight into future translational targets for therapy of heart failure. If lipid oxidation is largely enzyme-mediated, then specific enzymes can be targeted that will be identified in the proposed research. Alternatively, if lipid oxidation is largely mediated/initiated by non-enzymatic Fe+2 mechanisms then Fe chelation approaches and intramembrane radical traps can be explored. In Specific Aim 3, we will investigate the roles of oxidized lipids in the activation of different cell types from control and from failing hearts from female and male subjects. The importance of inflammation in heart disease is now well documented, but the roles oxidized lipids play in activating different cells of the immune system is still at its earliest stages of understanding. To traverse this gap in our knowledge, we will examine the effects of different oxidized lipids on selected cells in the cardiovascular system (e.g., cardiac myocytes, fibroblasts, endothelial cells, and macrophages). Through understanding the mechanisms underlying the deleterious effects of oxidized lipids on mitochondrial function in human hearts, a multi-tiered approach for treatment of congestive heart failure can be realized targeting enzyme-mediated oxidation, Fe2+ mediated oxidation and terminators of free radical propagation.

摘要 心血管疾病是工业化国家最常见的死亡原因。在我们学习的过程中， 我们已经确定了以前未发现的线粒体脂质代谢和信号通路，这些通路导致 2-花生四烯酰基-溶血磷脂酰胆碱(2-AA-LPC)和2-花生四烯酰基胆碱(2-AA-LPC) 溶血磷脂酰乙醇胺(2-AA-LPE)。这些包括：1)将iPLA2g(PNPLA8)鉴定为 具有sn-1特异性的磷脂酶；和2)氧化的心磷脂激活的细胞色素c作为血浆原酶。 催化合成2-AA LPC和2-AA LPE。进一步研究表明，2-AA-LPC和2-AA-LPE 是环氧合酶-2的极佳底物，导致大量意想不到的代谢物。值得注意的是， 将这些底物与12-脂氧合酶或15-脂氧合酶孵育，它们被氧化成12-脂氧合酶. H(P)ETE-或15-H(P)ETE-溶血磷脂。根据这些发现，我们确定了2-AA-LPC和2-AA-LPE 作为脂质合成和人类心脏线粒体功能中的信号和代谢节点。重要的是，我们有 确定了衰竭的人心脏线粒体在体内产生更多HETE二十烷基类化合物的能力 与非失败的对照线粒体相比，对钙离子挑战的反应。最近，铁下垂 被认为是通过积累脂质过氧化氢导致细胞死亡的一种机制。在心肌梗死期间 缺血和心力衰竭时，很大一部分线粒体铁(Fe+3)从结合状态释放出来，成为 通过Haber-Weiss和Fenton型引发活性氧物种(ROS)形成的游离Fe+2 化学药学。本研究的目的是确定主要机制对 氧化脂质及其在衰竭心肌膜功能障碍中的作用。拟议的研究将集中在 从机制上理解导致人体心脏衰竭时氧化脂质产生的机制。 具体地说，我们将识别不同类型的氧化脂质中的手性浓缩，以获得机理上的洞察 心力衰竭治疗的未来翻译目标。如果脂质氧化在很大程度上是由酶介导的，那么特异性 酶可以作为目标，将在拟议的研究中确定。或者，如果脂质氧化在很大程度上 非酶介导/引发的Fe+2机制、铁络合途径与膜内自由基 圈闭是可以勘探的。在具体目标3中，我们将研究氧化脂质在不同细胞激活中的作用。 类型来自对照和来自女性和男性受试者衰竭的心脏。心脏炎症的重要性 疾病现在已经有了很好的记载，但氧化脂质在激活免疫系统的不同细胞中所起的作用 仍处于理解的最早阶段。为了跨越我们知识中的这一鸿沟，我们将检查 心血管系统中特定细胞(如心肌细胞、成纤维细胞、内皮细胞)上的不同氧化脂质 细胞和巨噬细胞)。通过了解氧化脂质有害作用的潜在机制 在人类心脏线粒体功能方面，治疗充血性心力衰竭的多层次方法可以是 实现了靶向酶介导的氧化、Fe2+介导的氧化和自由基传播的终止物。