Oxygenated Species of Cardiolipins as Biomarkers of Mitochondrial Dysfunction

描述(申请人提供)：本申请的目标是确定与帕金森病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记物。这个团队已经证明了线粒体特有的磷脂，心磷脂(CL)，在神经元凋亡的早期经历了细胞色素c(Cytc)催化的选择性氧化。中心假设是，暴露在杀虫剂鱼藤酮下，导致CL时间和剂量依赖的选择性氧化，并通过在细胞凋亡早期触发的酶促细胞色素c催化反应，积累与线粒体功能障碍相关的氧化分子物种。CL分子物种的独特图谱代表了鱼藤酮诱导的与PD相关的线粒体功能障碍的一种新的生物标志物。利用氧化脂质组学方法，这组研究人员将首先确定鱼藤酮诱导的大鼠原代皮质和中脑神经元以及神经母细胞瘤SH-SY5Y中CL氧化分子物种的特定模式。此外，本研究的目的是利用大鼠鱼藤酮注射PD的模型，揭示多巴胺能神经元和皮质神经元中氧化的CL物种的特定轮廓。最后，在鱼藤酮作用下的人外周血淋巴细胞中存在鱼藤酮特异性CL氧化模式的一些建立将与在鱼藤酮输注的大鼠模型中检测到的结果进行比较。为了验证这一假设，我们开发了以下特定目标：特定目标1将利用氧化脂质组学来鉴定和表征原代大鼠皮质神经元、中脑神经元以及暴露于鱼藤酮的SH-SY5Y细胞中CL的分子种类和独特的立体特异性氧化产物。特异性目标2将建立细胞色素c与CL的相互作用参与原代培养的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。特定目的3将确定在体外神经元中检测和鉴定的鱼藤酮诱导的过氧化CL分子物种在体内注射鱼藤酮后在大脑皮层和中脑神经元线粒体中积累的程度。特定目标4将揭示人类外周血淋巴细胞中鱼藤酮特异性CL过氧化模式，作为与帕金森病相关的线粒体功能障碍的生物标志物。 公共卫生相关性：这项应用的目标是确定环境(鱼藤酮)诱导的线粒体功能障碍与帕金森病相关的生物标记物。这将通过一种新的氧化脂质组学方法实现。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的大鼠皮质和中脑神经元中被鉴定，并在人类外周血淋巴细胞中被揭示。

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.