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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）：线粒体功能随年龄增长而恶化。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活性降低相关，并与主要心磷脂种类[CL（C18：2）4]减少和含二十二碳六烯酸CL [CL（C18：2）3，（C22：6）1]增加相关。复合物IV的活性在磷脂的存在下恢复。这些与年龄相关的变化在肌纤维间（IFM）线粒体中观察到，但在肌膜下线粒体（SSM）中未观察到。由于线粒体电子传递链复合物的活性受到线粒体内膜脂质环境的深刻影响，特别是CL，我们提出CL周转/重塑增加。为了实验验证我们的假设，我们将使用一种新的重水（2 H2O）代谢标记技术，我们已经开发了一个全球的脂质体和蛋白质组营业额的研究在自由生活的动物。在目的1中，我们确定了老化对6和24月龄大鼠SSM和IFM中磷脂动力学的影响，特别关注了老年大鼠线粒体特异性的独特CL物质[CL（C18：2）3，（C22：6）1]的周转。CL的甘油部分将用于评估CL生物合成以及6月龄动物中次要非必需脂肪酸和24月龄动物中二十二碳六烯酸的周转。亚油酸是一种必需脂肪酸，不会被标记，这在两项已发表的研究中一直是一个问题。相比之下，我们建议使用甘油部分，其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和CIII结合到IFM中的超复合物中的CL的营业额和重塑，与SSM相比，其不受老化的影响。CL动力学在心脏IFM从老年大鼠恢复到SSM中观察到的和IFM为6个月大的动物后，恢复与乙酰肉毒碱的复合物III活性？在IFM中，特异性细胞色素B中的CIII蛋白的周转是否随着CIII活性和线粒体氧化磷酸化的恢复而增加？本R21的目的是通过使用我们为自由生活动物的全球蛋白质组周转研究开发的新型重水（2 H2O）代谢标记技术确定线粒体磷脂和CL的周转，收集与年龄相关的线粒体功能降低机制相关的初步数据。应用这种新的方法来评估衰老对线粒体功能的影响，将为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战略的发展，以应对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。