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。该团队已经证明，线粒体特异性磷脂，心磷脂（CL），在神经元凋亡的早期经历了细胞色素c （cyt c）催化的选择性氧化。核心假设是，暴露于农药鱼藤酮会引起时间和剂量依赖的CL选择性氧化，并通过细胞凋亡早期触发的酶促细胞c催化反应，导致其氧化分子物种的积累，从而导致线粒体功能障碍。CL分子种类的独特特征代表了鱼藤酮诱导的与帕金森病相关的线粒体功能障碍的一种新型生物标志物。利用氧化脂质组学方法，这组研究人员将首先确定鱼藤酮在大鼠初级皮质和中脑神经元以及神经母细胞瘤SH-SY5Y中诱导的CL氧化分子种类的特定模式。此外，目的是通过大鼠鱼藤酮输注PD模型揭示氧化CL物质在多巴胺能和皮质神经元中的特异性特征。最后，对暴露于鱼藤酮的人外周血淋巴细胞中存在鱼藤酮特异性CL氧化模式的一些研究将与在鱼藤酮输注大鼠模型中检测到的结果进行比较。具体目标1将利用氧化脂质组学来鉴定和表征大鼠皮质神经元和中脑神经元以及暴露于鱼tenone的SH-SY5Y细胞中CL的分子种类以及独特的立体特异性氧合产物。特异性目的2将建立cyt c与CL的相互作用参与大鼠皮质和中脑神经元以及鱼藤酮暴露的SH-SY5Y细胞CL氧化的机制和途径。特异性目的3将确定大鼠输注鱼藤酮后，在体外神经元中检测和鉴定的鱼藤酮诱导的过氧化物CL分子种在体内皮层和中脑神经元线粒体中积累的程度。特异性Aim 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中作为与帕金森病相关的线粒体功能障碍的生物标志物。

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.