Aged rat heart mitochondrial dysfunction: proteome and lipidome dynamics

老年大鼠心脏线粒体功能障碍：蛋白质组和脂质组动力学

基本信息

批准号：
8969074
负责人：
Charles Leslie Hoppel
金额：
$ 23.78万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-30 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8969074
关键词：
Accounting Acetylcarnitine Address Adult Affect Age Aging Anabolism Animals Binding Biochemical Cardiac Cardiolipins Cardiovascular Diseases Cardiovascular system Cell membrane Complex Cytochromes b Data Defect Deuterium Oxide Development Dinitrophenols Docosahexaenoic Acids Elderly Electron Transport Electron Transport Complex III Energy Metabolism Environment Essential Fatty Acids Fatty Acids Fostering Glues Glycerol Goals Heart Impairment Individual Injury Inner mitochondrial membrane Kinetics Knowledge Label Left Life Linoleic Acids Lipids Measures Membrane Metabolic Methods Minor Mitochondria Mitochondrial Proteins Modeling Molecular Morbidity - disease rate Myofibrils Organelles Oxidative Phosphorylation Phospholipids Production Proteins Proteome Public Health Publishing Rattus Replacement Therapy Reporting Research Respiration Risk Factors Site Structure System Techniques Testing Vertebral column Work age effect age related aged antioxidant therapy base complex IV heart metabolism in vivo inhibitor/antagonist insight mitochondrial dysfunction mortality novel novel strategies prevent public health relevance respiratory restoration tool

项目摘要

DESCRIPTION (provided by applicant): Mitochondrial function deteriorates with aging. We have shown that aging is associated with decreased mitochondrial protein, mitochondrial oxidative phosphorylation, complex III (CIII) and IV activity, and is associated with decrease in major cardiolipin species [CL(C18:2)4] and increase in docosahexaenoic acid containing CL [CL(C18:2)3, (C22:6)1]. The activity of complex IV is restored in the presence of phospholipids. These age-associated changes are observed in interfibrillar (IFM) mitochondria, but not in subsarcolemmal mitochondria (SSM). Because the activity of mitochondrial electron transport chain complexes is profoundly affected by the lipid environment of the mitochondrial inner membrane, specifically CL, we proposed that there is increased CL turnover/remodeling. To experimentally test our hypothesis we will use a novel heavy water (2H2O) metabolic labeling technique we have developed for a global lipidome and proteome turnover studies in free living animals. In Aim 1 we determine the effect of aging on phospholipid kinetics in SSM and IFM from 6 and 24 months old rats with particular focus on the turnover of the distinctive CL species [CL(C18:2)3, (C22:6)1] specific for mitochondria of aged rats. The glycerol moiety of CL will be used to assess CL biosynthesis and the turnover of minor nonessential fatty acids in the 6 month and docosahexaenoic acid in the 24 month old animals. Linoleic acid, an essential fatty acid, will not be labeled, which has been a problem in the two published studies. In contrast, we propose to use the glycerol moiety, which will be labeled and is the backbone while the fatty acids are part of the remodeling. In Aim 2 we determine the turnover and remodeling of CL associated with free CIII and CIII incorporated into supercomplexes in IFM in comparison to SSM, which are unaffected by aging. Is CL kinetics in cardiac IFM from aged rats restored to that observed in SSM and to IFM for 6 month old animals following restoration of complex III activity with acetyl-carnitine? Does the turnover of CIII protein, in specific cytochrome b, increase with restoration of CIII activity and of mitochondrial oxidative phosphorylation in IFM? The goal of this R21 is to collect preliminary data relevant to the mechanism of age-associated decreased mitochondrial function by determining the turnover of mitochondrial phospholipids and CL in specific using a novel heavy water (2H2O) metabolic labeling technique we have developed for a global proteome turnover studies in free living animals. Application of this novel approach to assess the effect of aging on mitochondrial function will provide valuable insight into the molecular mechanism of cardiac aging. The understanding gained from our proposed work will foster the development of strategies to counter the negative impact of aging on cardiac metabolism and to delay or slow the rate of cardiovascular mortality and morbidity.

描述（由申请人提供）：线粒体功能随衰老而恶化我们已经证明，衰老与线粒体蛋白、线粒体氧化磷酸化、复合物 III (CIII) 和 IV 活性的减少有关，并且与主要心磷脂种类 [CL() 的减少有关。 C18:2)4] 并增加含有 CL 的二十二碳六烯酸 [CL(C18:2)3, (C22:6)1]。在磷脂存在的情况下，复合物 IV 的活性会恢复。这些与年龄相关的变化在纤维间 (IFM) 线粒体中观察到，但在肌膜下线粒体 (SSM) 中观察不到，因为线粒体电子传递链复合物的活性深受脂质环境的影响。为了通过实验检验我们的假设，我们将使用我们开发的新型重水 (2H2O) 代谢标记技术。在自由活体动物中进行的全球脂质组和蛋白质组周转研究在目标 1 中，我们确定了衰老对 6 个月和 24 个月大大鼠的 SSM 和 IFM 中磷脂动力学的影响，特别关注独特的 CL 物种 [CL(C18) 的周转。 :2)3, (C22:6)1] 对老年大鼠的线粒体具有特异性 CL 的甘油部分将用于评估 CL 生物合成和次要非必需脂肪的周转。 6 个月大的动物中的脂肪酸和 24 个月大的动物中的二十二碳六烯酸（一种必需脂肪酸）不会被标记，这在两项已发表的研究中一直是一个问题，相反，我们建议使用甘油部分。它将被标记并且是主链，而脂肪酸是重塑的一部分。在目标 2 中，我们确定了与游离 CIII 相关的 CL 的周转和重塑，以及并入 IFM 中的超级复合物中的 CIII。与不受衰老影响的 SSM 相比，在用乙酰肉碱恢复复合物 III 活性后，老年大鼠心脏 IFM 中的 CL 动力学是否恢复到 SSM 和 IFM 中观察到的情况？，在特定细胞色素 b 中，随着 IFM 中 CIII 活性和线粒体氧化磷酸化的恢复而增加？该 R21 的目标是收集与年龄相关机制相关的初步数据。通过使用我们为自由活体动物的全球蛋白质组周转研究开发的新型重水（2H2O 减少）代谢标记技术来确定线粒体磷脂和 CL 的周转，从而评估线粒体功能，应用这种新方法来评估衰老的影响。对线粒体功能的研究将为了解心脏衰老的分子机制提供有价值的见解，从我们提出的工作中获得的理解将促进制定策略来对抗衰老对心脏代谢的负面影响，并延迟或减缓心血管死亡率和发病率。。