Quantitative Mitochondrial Proteomics of Healthy and Diabetic Mice

健康和糖尿病小鼠的定量线粒体蛋白质组学

• 批准号: 7937890
• 负责人: David J Pagliarini
• 金额: $ 43.72万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-25 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7937890
• 关键词: Address; Affect; Affinity Chromatography; Age; Area; Arts; Automobile Driving; Biological Markers; Cells; Communities; Coupled; Data; Development; Diabetes Mellitus; Diabetic mouse; Disease; Elements; Etiology; Exhibits; Functional disorder; Gene Expression; Genes; Genomics; Goals; Grant; Individual; Insulin; Insulin Resistance; Lipids; Maps; Mass Spectrum Analysis; Measures; Mediating; Messenger RNA; Metals; Mitochondria; Mitochondrial Proteins; Molecular; Mouse Strains; Mus; Muscle; Muscle Mitochondria; National Institute of Diabetes and Digestive and Kidney Diseases; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Onset of illness; Open Reading Frames; Organelles; Pancreas; Pathology; Patients; Pattern; Peripheral; Phosphoproteins; Phosphoric Monoester Hydrolases; Phosphotransferases; Play; Post-Translational Protein Processing; Proteins; Proteome; Proteomics; Reactive Oxygen Species; Relative (related person); Resistance; Resolution; Resources; Rest; Role; Signal Transduction; Signaling Protein; Site; Skeletal Muscle; Staging; System; Technology; Tissue Microarray; Tissues; Validation; base; design; diabetic; diabetic patient; fatty acid oxidation; glucose disposal; insulin secretion; interest; mitochondrial dysfunction; protein function; public health relevance; respiratory; therapy design

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (03): Biomarker Discovery and Validation, and specific Challenge Topic 03-DK-103: Identify the normal and diseased proteome of subcellular organelles of relevance to NIDDK diseases. Mitochondrial dysfunction has been postulated as a unifying theme of type 2 diabetes mellitus (T2DM), which currently affects more than 20 million individuals in the US alone. Decreased mitochondrial content and oxidative capacity, altered fatty acid oxidation, and a rise in mitochondria-derived reactive oxygen species have been associated with the development of insulin resistance and T2DM. However, the importance of these observations has recently been challenged, and it remains unclear which, if any, changes to mitochondrial composition and/or function actually contribute to T2DM etiology. The long-term objective of this proposal is to arrive at a fuller understanding of the importance of mitochondrial dysfunction in T2DM by elucidating the compositional changes that accompany the onset of this disease at unprecedented resolution. Despite the observations noted above, an understanding of the molecular basis for mitochondrial dysfunction in T2DM has largely been lacking for at least two reasons. First, until recently, much of the mammalian mitochondria proteome itself was undefined, making it impossible to conduct comprehensive comparisons of mitochondrial composition during different stages of this disease. Second, microarray analyses used to investigate global differences in gene expression between healthy and diabetic patients have been crippled by the markedly poor correlation between cellular mRNA and protein levels. Here, we propose to establish a comprehensive map of the proteomic and phosphoproteomic changes that occur during the onset of obesity-dependent T2DM using state-of-the-art quantitative proteomics (AIM1). To do so, we will take advantage of two mouse strains: C57BL/6 (B6) leptinob/ob mice, which are resistant to diabetes, and BTBR leptinob/ob mice, which develop severe diabetes as they age. During the two-year timeframe of this project, we will focus on mitochondria from skeletal muscle, the primary site of insulin- mediated glucose disposal in the body. This proteomic resource will also provide an opportunity to explore the mechanisms by which these mitochondrial alterations occur. In particular, we will use integrative genomics to elucidate the post-transcriptional and post-translational mechanisms at play in the control of mitochondrial gene expression and protein function (AIM2). Completion of our aims will clarify the mitochondrial restructuring in skeletal muscle that accompanies the onset of T2DM, provide a rich quantitative proteomic resource for the diabetes community, and lay the groundwork for identifying protein biomarkers and designing muscle-specific therapies targeted against this organelle to treat T2DM. PUBLIC HEALTH RELEVANCE: Mitochondrial dysfunction is a prominent feature of type 2 diabetes mellitus (T2DM), but the underlying basis for this dysfunction is not well understood. We propose to use state-of-the-art proteomics technologies to establish a map of mitochondrial alterations that occur in skeletal muscle during the onset of obesity-induced diabetes. Completion of this goal will help identify mitochondrial biomarkers for this disease, and provide a framework for designing therapies targeted against this organelle to treat T2DM.

描述（由申请人提供）：本申请涉及广泛的挑战领域 (03)：生物标志物发现和验证，以及具体的挑战主题 03-DK-103：识别与 NIDDK 疾病相关的亚细胞细胞器的正常和患病蛋白质组。线粒体功能障碍被认为是 2 型糖尿病 (T2DM) 的一个统一主题，目前仅在美国就有超过 2000 万人受到影响。线粒体含量和氧化能力的降低、脂肪酸氧化的改变以及线粒体衍生的活性氧的增加与胰岛素抵抗和 T2DM 的发展有关。然而，这些观察结果的重要性最近受到了挑战，目前尚不清楚线粒体组成和/或功能的哪些变化（如果有的话）实际上导致了 T2DM 病因。该提案的长期目标是通过以前所未有的分辨率阐明伴随这种疾病发作的成分变化，从而更全面地了解线粒体功能障碍在 T2DM 中的重要性。尽管有上述观察结果，但由于至少两个原因，人们对 T2DM 线粒体功能障碍的分子基础仍缺乏了解。首先，直到最近，哺乳动物线粒体蛋白质组本身的大部分尚不清楚，因此无法对该疾病不同阶段的线粒体组成进行全面比较。其次，用于研究健康患者和糖尿病患者之间基因表达整体差异的微阵列分析因细胞 mRNA 和蛋白质水平之间的相关性明显较差而受到削弱。在这里，我们建议使用最先进的定量蛋白质组学 (AIM1) 建立肥胖依赖性 T2DM 发病期间发生的蛋白质组和磷酸化蛋白质组变化的综合图谱。为此，我们将利用两种小鼠品系：C57BL/6 (B6) leptinob/ob 小鼠（对糖尿病有抵抗力）和 BTBR leptinob/ob 小鼠（随着年龄的增长，会出现严重的糖尿病）。在该项目的两年时间内，我们将重点关注骨骼肌的线粒体，这是体内胰岛素介导的葡萄糖处理的主要部位。这种蛋白质组学资源还将提供一个探索这些线​​粒体改变发生机制的机会。特别是，我们将使用整合基因组学来阐明在控制线粒体基因表达和蛋白质功能（AIM2）中发挥作用的转录后和翻译后机制。我们的目标的完成将阐明伴随 T2DM 发病的骨骼肌线粒体重组，为糖尿病界提供丰富的定量蛋白质组学资源，并为识别蛋白质生物标志物和设计针对该细胞器的肌肉特异性疗法来治疗 T2DM 奠定基础。 公共卫生相关性：线粒体功能障碍是 2 型糖尿病 (T2DM) 的一个突出特征，但这种功能障碍的根本原因尚不清楚。我们建议使用最先进的蛋白质组学技术来建立肥胖引起的糖尿病发作期间骨骼肌中发生的线粒体改变图谱。这一目标的完成将有助于识别该疾病的线粒体生物标志物，并为设计针对该细胞器的疗法来治疗 T2DM 提供框架。