Mitochondrial Dysfunction and Adipose Insulin Resistance

线粒体功能障碍和脂肪胰岛素抵抗

• 批准号: 8531229
• 负责人: David A Bernlohr
• 金额: $ 29.98万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8531229
• 关键词: Adipocytes; Adipose tissue; Animal Model; Animals; Antioxidants; Biology; Cardiovascular Diseases; Cell Culture Techniques; Complex; Development; Diet; Disease; Disease model; Down-Regulation; Electron Transport; Enzymes; Etiology; Event; Fatty acid glycerol esters; Genetic Determinism; Genomics; Glutathione S-Transferase; Hormonal; Human; Human Biology; Hydrogen Peroxide; Hydroxyl Radical; Hypertension; Individual; Insulin; Insulin Resistance; Laboratories; Laboratory Study; Lead; Link; Lipids; MAPK14 gene; Metabolic; Metabolic Diseases; Metabolism; Minnesota; Mitochondria; Mitochondrial Proteins; Modeling; Modification; Molecular; N-terminal; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Oxidative Phosphorylation; Oxidative Stress; Pathway interactions; Phosphotransferases; Process; Production; Proteins; Proteomics; Reactive Oxygen Species; Reporting; Risk Factors; Role; Signal Transduction; Staging; Sum; Superoxides; System; Systems Biology; Technology; Testing; Thioredoxin; Thioredoxin-2; Transgenic Mice; Tumor Necrosis Factor-alpha; Universities; Work; combat; gain of function; insight; insulin sensitivity; jun Oncogene; metabolomics; mitochondrial dysfunction; novel; oxidation; public health relevance

DESCRIPTION (provided by applicant): A variety of animal, cell culture and molecular studies have correlated increased oxidative stress and the accumulation of reactive oxygen species (ROS) such as superoxide anion and hydroxyl radicals to type 2 diabetes. While appreciated for years, it was unclear if oxidative stress / ROS was a casual or causal factor in the etiology of the disease. However, more recent work utilizing a variety of loss and gain of function analyses have indicated that oxidative stress is causally linked to insulin resistance but that the molecular mechanisms remain obscure. This application will profile new and compelling findings from our laboratories using animal and cell culture models that establish a molecular linkage between the antioxidant defense system in adipose cells with mitochondrial function, oxidative phosphorylation, signal transduction and the development of insulin resistance. We present a novel hypothesis supported by preliminary studies from genomic, proteomic, metabolomic and molecular analyses that point toward the covalent modification of mitochondrial proteins with bioactive lipids and the oxidation of mitochondrial thioredoxin as central to the process. Initiating this oxidative stress challenge are new findings that describe the tumor necrosis factor 1 (TNF1) dependent down regulation of glutathione S-transferase A4 setting the stage for a molecular cascade of events that activates the c-JUN NH2-terminal kinase (JNK), an established regulator of insulin sensitivity. Moreover, we present new findings that demonstrate that the down regulation of GSTA4 is not merely a process observed in animal models of insulin resistance but also occurs selectively in obese, insulin resistant, but not obese, insulin sensitive humans thereby providing a molecular differentiation between obesity and insulin resistance. This application builds on recent evidence obtained in the Bernlohr, Griffin and Arriaga laboratories that functionally links oxidative stress to insulin resistance. These studies in sum lead to our central hypothesis: decreased expression of GSTA4 in adipocytes leads to increased carbonylation of multiple protein targets. Carbonylation in turn initiates a cascade of molecular events leading to mitochondrial dysfunction and ROS production. ROS production leads to the oxidation of Thioredoxin 2 (Trx2) and the activation of Trx2-ASK1-JNK/p38 signaling system contributing to insulin resistance. To test this hypothesis, the following four specific aims are proposed: Specific Aim 1. Evaluate mitochondrial protein carbonylation and identify target proteins. Specific Aim 2. Assess ROS production and mitochondrial electron transport system in cell culture and animal models. Specific Aim 3. Develop and characterize aP2-HA-GSTA4 transgenic mice maintained on low and high fat diets. Specific Aim 4. Characterize cellular metabolism and the Trx2-ASK1-JNK pathway in animal and cell culture models. PUBLIC HEALTH RELEVANCE: A variety of animal, cell culture and molecular studies have correlated increased oxidative stress and the accumulation of reactive oxygen species (ROS) to type 2 diabetes. While appreciated for years, it was unclear if oxidative stress / ROS was a casual or causal factor in the etiology of the disease. This application will profile new and compelling findings from our laboratories using animal and cell culture models that establish a molecular linkage between the antioxidant defense system in adipose cells with mitochondrial function, oxidative phosphorylation, signal transduction and the development of insulin resistance. We present new findings that demonstrate that the down regulation of GSTA4 is not merely a process observed in animal models of insulin resistance but also occurs selectively in obese, insulin resistant, but not obese, insulin sensitive humans thereby providing a molecular differentiation between obesity and insulin resistance. If the hypothesis is proven to be correct, the study would be immediately translatable to human biology and afford a new view of how type 2 diabetes may be combated.

描述（由申请人提供）：各种动物、细胞培养和分子研究已将氧化应激增加和活性氧（ROS）（如超氧阴离子和羟基自由基）积累与2型糖尿病相关。虽然多年来一直受到重视，但尚不清楚氧化应激/ ROS是否是该疾病病因学中的偶然或因果因素。然而，最近的工作利用各种损失和获得的功能分析表明，氧化应激与胰岛素抵抗的因果关系，但分子机制仍然模糊。该应用程序将介绍我们实验室使用动物和细胞培养模型的新的和令人信服的发现，这些模型在脂肪细胞中的抗氧化防御系统与线粒体功能，氧化磷酸化，信号转导和胰岛素抵抗的发展之间建立了分子联系。我们提出了一个新的假设，支持初步研究，从基因组学，蛋白质组学，代谢组学和分子分析，指向共价修饰的线粒体蛋白质与生物活性脂质和氧化的线粒体硫氧还蛋白作为中央的过程。启动这种氧化应激挑战的是新的发现，描述了肿瘤坏死因子1（TNF 1）依赖的谷胱甘肽S-转移酶A4的下调，为激活c-JUN NH 2-末端激酶（JNK）的分子级联事件奠定了基础，JNK是胰岛素敏感性的既定调节因子。此外，我们提出了新的发现，证明GSTA 4的下调不仅是在胰岛素抵抗的动物模型中观察到的过程，而且还选择性地发生在肥胖的胰岛素抵抗的人中，而不是肥胖的胰岛素敏感的人中，从而提供了肥胖和胰岛素抵抗之间的分子差异。这项应用建立在Bernlohr，Griffin和Arriaga实验室最近获得的证据基础上，这些证据在功能上将氧化应激与胰岛素抵抗联系起来。总之，这些研究导致了我们的中心假设：脂肪细胞中GSTA 4表达减少导致多个蛋白质靶点的羰基化增加。羰基化反过来启动一系列分子事件，导致线粒体功能障碍和ROS产生。ROS的产生导致硫氧还蛋白2（Trx 2）的氧化和Trx 2-ASK 1-JNK/p38信号系统的激活，从而导致胰岛素抵抗。为了验证这一假设，提出了以下四个具体目标：具体目标1。评估线粒体蛋白质羰基化并鉴定靶蛋白。具体目标2。在细胞培养和动物模型中评估ROS产生和线粒体电子传递系统。具体目标3。开发和表征维持在低脂肪和高脂肪饮食下的aP 2-HA-GSTA 4转基因小鼠。具体目标4。在动物和细胞培养模型中表征细胞代谢和Trx 2-ASK 1-JNK通路。 公共卫生关系：各种动物、细胞培养和分子研究已将氧化应激增加和活性氧（ROS）积累与2型糖尿病相关。虽然多年来一直受到重视，但尚不清楚氧化应激/ ROS是否是该疾病病因学中的偶然或因果因素。该应用程序将介绍我们实验室使用动物和细胞培养模型的新的和令人信服的发现，这些模型在脂肪细胞中的抗氧化防御系统与线粒体功能，氧化磷酸化，信号转导和胰岛素抵抗的发展之间建立了分子联系。我们提出了新的发现，证明GSTA 4的下调不仅是在胰岛素抵抗的动物模型中观察到的过程，而且选择性地发生在肥胖、胰岛素抵抗但不肥胖、胰岛素敏感的人中，从而提供了肥胖和胰岛素抵抗之间的分子差异。如果这一假设被证明是正确的，这项研究将立即转化为人类生物学，并为如何对抗2型糖尿病提供新的观点。