Oxygenated Species of Cardiolipins as Biomarkers of Mitochondrial Dysfunction

心磷脂的含氧物质作为线粒体功能障碍的生物标志物

• 批准号: 8334604
• 负责人: Valerian E Kagan
• 金额: $ 34.09万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-19 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8334604
• 关键词: Affect; Age; Apoptosis; Apoptotic; Biological Markers; Brain; Cardiolipins; Cells; Cessation of life; Characteristics; Chromatography; Cytosol; Development; Diagnosis; Disease; Dopamine; Dose; Exposure to; Fatty Acids; Fingers; Genes; Goals; Heating; Human; Impairment; In Vitro; Incidence; Individual; Infusion procedures; Link; Liquid Chromatography; Maps; Midbrain structure; Mitochondria; Modeling; Modification; Molecular; Mutate; Nature; Neuroblastoma; Neurodegenerative Disorders; Neurons; Outcome; Oxidative Stress; Parkinson Disease; Pathway interactions; Patients; Pattern; Peripheral Blood Lymphocyte; Pesticides; Phospholipids; Polyunsaturated Fatty Acids; Population; Printing; Protocols documentation; Rattus; Reaction; Reactive Oxygen Species; Resolution; Role; Rotenone; Small Interfering RNA; Substantia nigra structure; System; Techniques; Testing; Time; United States; Work; base; cardiolipin synthase; cytochrome c; in vivo; mitochondrial dysfunction; neuron apoptosis; novel; oxidation; peroxidation

The goal of this application is to identify phospholipid biomarkers of environmentally-induced (rotenone- induced) mitochondrial dysfunction associated with Parkinson's disease PD. We demonstrated that a mitochondria specific phospholipid, cardiolipin (CL), undergoes selective oxidation catalyzed by cytochrome c (cyt c) early during neuronal apoptosis. Our central hypothesis is that exposure to a pesticide, rotenone, causes time- and dose-dependent selective oxidation of CL and accumulation of its oxidized molecular species associated with mitochondrial dysfunction through enzymatic cyt c catalyzed reactions triggered early in apoptosis. The unique profile of CL molecular species represents a new type of biomarkers of rotenone-induced mitochondrial dysfunction associated with PD. Using oxidative lipidomics approach we will first identify specific patterns of CL oxidized molecular species induced in rat primary cortical and midbrain neurons as well as neuroblastoma SH-SY5Y by rotenone. Further, we will reveal the specific profiles of oxidized CL species in dopaminergic and cortical neurons using rat rotenone- infusion model of PD. Finally we will establish the presence of the rotenone-specific CL oxidation patterns in human peripheral blood lymphocytes exposed to rotenone and compare them with those detected in rotenone-infusion rat model. The following Specific Aims were developed to test the hypothesis: Specific Aim 1 will utilize oxidative lipidomics to identify and characterize molecular species of CL as well as unique stereo-specific oxygenated products of CL in primary rat cortical neurons and midbrain neurons as well as in SH-SY5Y cells upon exposure to rotenone. Specific Aim 2 will establish the mechanisms and pathways through which interactions of cyt c with CL are involved in CL oxidation in primary rat cortical and midbrain neurons as well as SH-SY5Y cells exposed to rotenone. Specific Aim 3 will determine the extent to which molecular species of rotenone-induced peroxidized CL detected and identified in neurons in vitro accumulate in mitochondria of cortical and midbrain neurons in vivo after infusion of rotenone to rats. Specific Aim 4 will reveal rotenone-specific CL peroxidation patterns in human peripheral blood lymphocytes as biomarkers of mitochondrial dysfunction associated with PD.

本申请的目标是鉴定环境诱导的（鱼藤酮-）磷脂生物标志物。 诱导的）与帕金森病PD相关的线粒体功能障碍。我们证明， 线粒体特异性磷脂心磷脂（CL）在细胞色素c催化下发生选择性氧化 (cyt c）神经元凋亡早期。我们的中心假设是接触杀虫剂鱼藤酮， 导致CL的时间和剂量依赖性选择性氧化和其氧化的蓄积 与通过酶促细胞色素C催化的线粒体功能障碍相关的分子种类 在凋亡早期触发的反应。CL分子种类的独特概况代表了一种新的 鱼藤酮诱导的与PD相关的线粒体功能障碍的生物标志物类型。使用氧化 脂质组学方法，我们将首先确定大鼠中诱导的CL氧化分子种类的特定模式， 鱼藤酮对原代皮层和中脑神经元以及神经母细胞瘤SH-SY 5 Y的作用。此外，我们将 揭示了使用大鼠鱼藤酮在多巴胺能和皮质神经元中氧化CL物质的特异性特征， PD输注模型。最后，我们将建立鱼藤酮特异性CL氧化模式的存在， 人外周血淋巴细胞暴露于鱼藤酮，并与 鱼藤酮灌注大鼠模型。为了检验假设，制定了以下具体目标： 1将利用氧化脂质组学来鉴定和表征CL的分子种类， CL在原代大鼠皮层神经元和中脑中的独特立体特异性氧合产物 神经元以及SH-SY 5 Y细胞暴露于鱼藤酮。具体目标2将建立 细胞色素c与CL相互作用参与CL氧化的机制和途径， 原代大鼠皮质和中脑神经元以及暴露于鱼藤酮的SH-SY 5 Y细胞。具体 目的3将确定鱼藤酮诱导的过氧化CL的分子种类 在体外神经元中检测和鉴定在皮质和中脑线粒体中积累 鱼藤酮灌注大鼠后，体内神经元。Specific Aim 4将揭示鱼藤酮特异性CL 人外周血淋巴细胞线粒体过氧化模式 与PD相关的功能障碍。