Oxygenated Species of Cardiolipins as Biomarkers of Mitochondrial Dysfunction

描述（由申请人提供）：本申请的目的是鉴定与帕金森病PD相关的环境诱导（鱼藤酮诱导）线粒体功能障碍的磷脂生物标志物。这个研究小组已经证明，线粒体特异性磷脂，心磷脂（CL），经历细胞色素c（cyt c）在神经元凋亡早期催化的选择性氧化。中心假设是，暴露于农药鱼藤酮导致CL的时间和剂量依赖性选择性氧化和其氧化分子种类的积累与线粒体功能障碍相关，通过细胞色素c酶催化的反应在细胞凋亡早期触发。CL分子种类的独特谱代表了与PD相关的鱼藤酮诱导的线粒体功能障碍的新型生物标志物。使用氧化脂质组学方法，该组研究人员将首先确定鱼藤酮在大鼠原代皮层和中脑神经元以及神经母细胞瘤SH-SY 5 Y中诱导的CL氧化分子种类的特定模式。此外，本研究的目的是使用大鼠鱼藤酮灌注PD模型揭示多巴胺能和皮质神经元中氧化CL物质的特异性谱。最后，一些建立鱼藤酮特异性CL氧化模式暴露于鱼藤酮的人外周血淋巴细胞的存在下，将与鱼藤酮输注大鼠模型中检测到的那些进行比较。开发了以下特定目的以检验假设：特定目的1将利用氧化脂质组学来鉴定和表征暴露于鱼藤酮后原代大鼠皮质神经元和中脑神经元以及SH-SY 5 Y细胞中CL的分子种类以及CL的独特立体特异性含氧产物。具体目标2将建立机制和途径，通过这些机制和途径，在原代大鼠皮层和中脑神经元以及暴露于鱼藤酮的SH-SY 5 Y细胞中，cyt c与CL的相互作用参与CL氧化。具体目标3将确定在何种程度上鱼藤酮诱导的过氧化CL的分子种类检测和识别的神经元在体外积累的皮质和中脑神经元的线粒体在体内输注鱼藤酮给大鼠后。具体目标4将揭示鱼藤酮特异性CL过氧化模式在人外周血淋巴细胞作为线粒体功能障碍与PD相关的生物标志物。 公共卫生相关性：本申请的目的是鉴定与帕金森病相关的环境（鱼藤酮）诱导的线粒体功能障碍的生物标志物。这将通过一种新的氧化脂质组学方法来实现。线粒体心磷脂的鱼藤酮特异性过氧化模式将在体外和体内大鼠皮质和中脑神经元中确定，并在人外周血淋巴细胞中发现。

DESCRIPTION (provided by applicant): The goal of this application is to identify phospholipid biomarkers of environmentally-induced (rotenone- induced) mitochondrial dysfunction associated with Parkinson's disease PD. This team has demonstrated that a mitochondria specific phospholipid, cardiolipin (CL), undergoes selective oxidation catalyzed by cytochrome c (cyt c) early during neuronal apoptosis. The central hypothesis is that exposure to the 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 this group of investigators 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, the intent is to reveal the specific profiles of oxidized CL species in dopaminergic and cortical neurons using rat rotenone- infusion model of PD. Finally, some establishment for the presence of rotenone-specific CL oxidation patterns in human peripheral blood lymphocytes exposed to rotenone will be compared with those detected in the 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. PUBLIC HEALTH RELEVANCE: The goal of this application is to identify biomarkers of environmentally (rotenone)-induced mitochondrial dysfunction associated with Parkinson's Disease. This will be achieved by a novel oxidative lipidomics approach. Rotenone-specific peroxidation patterns of mitochondrial cardiolipins will be identified in rat cortical and midbrain neurons in vitro and in vivo and revealed in human peripheral blood lymphocytes.