HNE damage of adenine nucleotide translocase in ethanol-mediated neuron apoptosis

乙醇介导的神经元凋亡中腺嘌呤核苷酸转位酶的 HNE 损伤

• 批准号: 7934507
• 负责人: GEORGE I HENDERSON
• 金额: $ 21.46万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-20 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7934507
• 关键词: 4 hydroxynonenal; Address; Adenine Nucleotide Translocase; Aldehydes; Amino Acids; Apoptosis; Apoptotic; Area; Binding; Brain; Cell Death; Cell Membrane Permeability; Cerebrum; Cessation of life; Clinical; Complex; Cyclophilins; Data; Development; Ethanol; Event; Experimental Models; Extravasation; Family; Induction of Apoptosis; Inner mitochondrial membrane; Intervention; Lead; Literature; Mass Spectrum Analysis; Mediating; Mitochondria; Modification; Necrosis; Neurons; Oxidative Stress; Permeability; Play; Rattus; Regulation; Role; Side; Sulfhydryl Compounds; Treatment Protocols; adduct; alcohol response; alcohol sensitivity; antiporter; clinically relevant; crosslink; cyclophilin D; fetal; insight; member; mitochondrial permeability transition pore; neuron apoptosis; novel; pro-apoptotic protein; protein folding; public health relevance; research study; response

DESCRIPTION (provided by applicant): The overall hypothesis of this exploratory proposal is that a key mechanism underlying ethanol-mediated opening of the mitochondrial permeability transition pore complex and possibly apoptosis activation, is perturbation of adenine nucleotide translocase (ANT) function by 4- hydroxynonenal (HNE) adduct formation. Key responses to HNE are, as previously documented, inhibition of ANT activity, activation of the mitochondrial permeability transition pore, and ultimate release of pro-apoptotic proteins. It is proposed that this occurs as a multifactorial event which includes direct modification of specific amino acid residues of ANT and its impaired interactions with cyclophilin D (CyP-D). Significance: This proposal address a new and novel, mitochondrially-mediated, fundamental mechanism by which ethanol may elicit apoptotic death of neurons. Neuron survival control points such as this are prime targets for the ultimate development of new clinical interventions. Specific Aim 1 will address the hypothesis that modifications of adenine nucleotide translocase by 4- hydroxynonenal adduct formation is a mechanism underlying ethanol-related effects on the mitochondrial permeability transition pore complex and ultimately on induction of apoptosis. Specific Aim 2 will address the hypothesis that direct modifications of cyclophilin-D by 4- hydroxynonenal adduct formation alter critical interactions with adenine nucleotide translocase and that this is one mechanism underlying ethanol-related effects on the mitochondrial permeability transition pore complex. Experimental Approaches: The experimental approaches will utilize Mass Spectrometry to determine specific amino acid residues on ANT and CyP-D that are modified and point-mutational manipulations to establish the functional significance of these ethanol-related modifications. Studies will also establish effects of these specific ANT and CyP-D modifications on CyP-D binding to ANT, on permeability transition regulation, and on sensitivity to ethanol-mediated neuron death. The experimental model will be cultured fetal rat cerebral cortical neurons exposed to clinically relevant ethanol treatment regimens. PUBLIC HEALTH RELEVANCE: Exposure of cultured fetal rat cerebral cortical neurons to ethanol rapidly elicits oxidative stress and increased formation of 4-hydroxynonenal (HNE) in mitochondria, which is followed by mitochondrial leakage of pro-apoptotic proteins and apoptotic death. This exploratory proposal addresses the hypothesis that a mechanism underlying ethanol-mediated opening of the permeability transition pore complex (PTPC) is connected to HNE modifications of the pore components, adenine nucleotide translocase (ANT) and cyclophilin D (CyP-D). Thus, the proposed studies will provide insight into basic mechanisms underlying mitotoxic responses to ethanol as well as fundamental mechanisms of PTPC function.

描述（由申请人提供）：该探索性建议的总体假设是，乙醇介导的线粒体通透性过渡孔复合物的开放且可能是凋亡激活的关键机制，是腺嘌呤核苷酸易生酶（ANT）功能的扰动，是由4-羟基诺烯烯醇（HNE）添加（HNE）形成的。如前所述，对HNE的关键反应是对ANT活性的抑制作用，线粒体通透性过渡孔的激活以及促凋亡蛋白的最终释放。有人提出，这是一个多因素事件，其中包括对蚂蚁的特定氨基酸残基的直接修饰及其与环磷脂D（CYP-D）的相互作用受损。意义：该提案介绍了一种新的新型，线粒体介导的基本机制，乙醇可能引起神经元的凋亡死亡。诸如此类的神经元存活控制点是新临床干预措施最终发展的主要目标。具体目标1将解决以下假设：4-羟基烯醇加合物形成对腺嘌呤核苷酸易位酶的修饰是一种与乙醇相关的机制对乙醇相关的作用，对线粒体渗透性过渡孔复合物，最终对凋亡的诱导最终。具体目标2将解决以下假设：通过4-羟基诺纳尔加合物形成直接修饰环磷脂-D，改变了与腺嘌呤核苷酸易位酶的关键相互作用，这是对乙醇相关的一种机制对线粒体通透性过渡性过渡孔复合物的一种机制。实验方法：实验方法将利用质谱法来确定经过修饰的ANT和CYP-D上的特定氨基酸残基，并确定点突破性操作来确定这些与乙醇相关的修饰的功能意义。研究还将建立这些特定的蚂蚁和CYP-D修饰对CYP-D结合，渗透性转变调节以及对乙醇介导的神经元死亡的敏感性的影响。实验模型将是暴露于临床相关乙醇治疗方案的胎儿大鼠脑皮质神经元。 公共卫生相关性：培养的胎儿大鼠脑皮质神经元暴露于乙醇中，会迅速引起氧化应激，并增加线粒体中4-羟基诺烯（HNE）的形成，随后是线粒体渗漏的促脑蛋白质蛋白质和凋亡性死亡。该探索性提议解决了以下假设：乙醇介导的渗透性过渡孔复合物（PTPC）的基础机制与孔成分的HNE修饰有关，腺嘌呤核苷酸转换酶（ANT）和环醇D（CYP-D）连接。因此，拟议的研究将洞悉对乙醇的丝毒反应以及PTPC功能的基本机制。