SIRT1 Overexpression in Cellular Mitochondrial Metabolism and Function

SIRT1 在细胞线粒体代谢和功能中的过表达

• 批准号: 7449824
• 负责人: JAMES W RUSSELL
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7449824
• 关键词: Adipocytes; Aerobic; Affect; Aging; Alzheimer' s Disease; Anatomy; Animal Model; Antioxidants; Biological; Biology; Brain; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Caloric Restriction; Cardiac; Cardiomyopathies; Cardiovascular system; Cell Respiration; Cells; Cerebrovascular Disorders; Complications of Diabetes Mellitus; Condition; Coupled; Data; Deacetylation; Defect; Development; Diabetes Mellitus; Diabetic Neuropathies; Diet; Disease; Disease model; Doxycycline; Dyslipidemias; Electron Transport; Elements; Energy Metabolism; Enzymes; Failure; Family; Gene Expression; Generations; Genes; Grapes; Health; Health Benefit; Heart; Human; Hyperglycemia; Incidence; Injury; Institutes; Insulin Resistance; Knockout Mice; Laboratories; Life; Link; Longevity; Measures; Mediating; Membrane; Metabolic Diseases; Metabolism; Mitochondria; Modeling; Molecular; Mus; Muscle; National Center for Research Resources; National Institute of Diabetes and Digestive and Kidney Diseases; Natural regeneration; Nerve Degeneration; Nervous System Physiology; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neurologic; Neuronal Injury; Neurons; Nicotinamide adenine dinucleotide; Nuclear; Numbers; Obesity; Obesity associated disease; Oxidative Phosphorylation; Oxidative Stress; Pancreas; Parkinson Disease; Pathology; Pathway interactions; Peripheral; Peripheral Nerves; Peripheral Nervous System; Peroxisome Proliferator-Activated Receptors; Phenotype; Physical activity; Play; Prevention strategy; Process; Prosencephalon; Protein Overexpression; Proteins; Public Health; Range; Regulation; Research; Research Personnel; Resveratrol; Risk; Role; Sirtuins; Skeletal Muscle; Skin; Testing; Tetanus Helper Peptide; Tetracycline; Tetracycline Control; Tetracyclines; Therapeutic; Tissues; Toxic effect; Trans-Activators; Transgenic Mice; United States National Institutes of Health; Up-Regulation; Yeasts; age effect; aging gene; aging population; diabetic; human disease; human study; improved; interest; member; mitochondrial dysfunction; mouse model; novel; nrf1 protein; prevent; promoter; repaired; respiratory; response; transcription factor; young adult

DESCRIPTION (provided by applicant): Oxidative stress and mitochondrial dysfunction have been associated with a wide range of neurodegenerative diseases and metabolic disorders such as diabetes. A major public health problem is the increase in the incidence of obesity-related diseases, such as diabetes and its complications and the increased incidence of neurodegenerative diseases, for example Parkinson's and Alzheimer's diseases in the aging population. Therapeutic and preventive strategies to reduce the complications of diabetes and to treat neurodegenerative diseases are urgently needed. Several lines of evidence indicate that a common link in these diseases is diminished mitochondrial oxidative phosphorylation and response to oxidative injury. Key regulators of mitochondrial function, the nuclear respiratory coactivators help to regulate mitochondrial oxidative phosphorylation and prevent cellular and neuronal injury. SIRT1 is a member of the sirtuin family of NAD+dependent deacetylases, which is proposed to be responsible for health benefits provided by caloric restriction. Furthermore, resveratrol found in the skin of red grapes increases the activity SIRT1, prolongs life-span in mice, and may prevent neurodegeneration. A key component of the protective response mediated by SIRT1 is deacetylation and activation of the transcription factor PGC-11 leading to increased mitochondrial regeneration and improved cellular oxidative energy metabolism. The role of SIRT1 and its mechanism/s of action at cellular level are uncertain; however investigators with a wide spectrum of research foci have an interest in understanding the biological actions of SIRT1 in different tissues. To study this, generation of transgenic mice that conditionally expresses SIRT1 is needed. In response to an PA from NCRR (PA-07-336) to develop animal models of human disease that are applicable to the research interests of two or more categorical NIH Institutes/Centers, we will develop a transgenic mouse that expresses mouse SIRT1 and mitochondrial targeted enhanced yellow fluorescent protein under the control of tetracycline responsive element (TRE-SIRT1/mito-eYFP). Co-expression of mito-eYFP with SIRT1 will be used to identify, isolate and study the influence of SIRT1 expression on mitochondrial function. Then, SIRT1 expression will be targeted to central and peripheral neurons by crossing it with CamKII-1 tTA mice. The transgenic mice developed in this study will aid investigators from NINDS, NIDDK, NIA and other institutes to test disease mechanisms and develop SIRT1 mediated therapies. Specifically in this proposal, we will investigate the mechanism by which SIRT1 protects central and peripheral neurons against diabetes-induced neuronal injury. The proposal also describes how the animal models produced can be developed by other investigators to study SIRT1 biology in non-neuronal cells. We have two aims: (1) To develop a transgenic mouse that expresses mouse SIRT1 under the control of tetracycline responsive element (TRE;TRE-SIRT1/mito-eYFP). (2) To phenotype the bigenic SIRT1 neuron specific mouse model. Lay description: Obesity, diabetes, and neurodegenerative diseases affect large numbers of people. The SIRT1 protein is considered to be a master regulator of the body's defense against disease and is activated by resveratrol found in red grapes. We aim to uncover the mechanism by which S1RT1 protects neurons from diabetes induced neurological complications. PUBLIC HEALTH RELEVANCE: This proposal is submitted in response to a PA from NCRR PA-07-336 "Development of Animal models and Related Biological Materials for Research". The research objective of this PA is to "develop, characterize or improve animal models for human disease and that models to be considered must be applicable to the research interests of two or more categorical NIH Institutes/Centers". A major public health problem is the increase in the incidence of obesity-related diseases, such as diabetes and its complications and the increased incidence of neurodegenerative diseases, for example Parkinson's and Alzheimer's diseases in the aging population. We will develop a transgenic mouse that expresses mouse SIRT1 and mitochondrial targeted enhanced yellow fluorescent protein under the control of tetracycline responsive element (TRE-SIRT1/mito-eYFP). Co-expression of mito-eYFP with SIRT1 will be used to identify, isolate and study the influence of SIRT1 expression on mitochondrial function. Then, SIRT1 expression will be targeted to central and peripheral neurons by crossing it with CamKII-1 tTA mice. The transgenic mice developed in this study will aid investigators from NINDS, NIDDK, NIA and other institutes to test disease mechanisms and develop SIRT1 mediated therapies. Our overall hypothesis is that activation of SIRT1 in the central (CNS) and peripheral nervous system (PNS) would reduce oxidative stress and improve regulation of Mt function in neurons and other tissues that may be important in neurodegenerative diseases, diabetes and its complications, and in delaying or reducing the effect of aging in the nervous system. Activation of the sirtuins offers the potential for a novel treatment of several human diseases that are related to oxidative injury and defects of mitochondrial function.

描述（由申请人提供）：氧化应激和线粒体功能障碍与广泛的神经退行性疾病和代谢紊乱（如糖尿病）相关。一个主要的公共卫生问题是与肥胖有关的疾病的发病率增加，例如糖尿病及其并发症，以及神经变性疾病的发病率增加，例如老年人中的帕金森病和阿尔茨海默病。迫切需要减少糖尿病并发症和治疗神经退行性疾病的治疗和预防策略。一些证据表明，这些疾病的共同联系是线粒体氧化磷酸化和对氧化损伤的反应减少。线粒体功能的关键调节因子，核呼吸辅激活因子有助于调节线粒体氧化磷酸化并防止细胞和神经元损伤。SIRT 1是NAD+依赖性脱乙酰酶的sirtuin家族的成员，其被认为负责热量限制提供的健康益处。此外，在红葡萄皮中发现的白藜芦醇增加了SIRT 1的活性，延长了小鼠的寿命，并可能防止神经退行性疾病。SIRT 1介导的保护性反应的一个关键组成部分是转录因子PGC-11的脱乙酰化和活化，导致线粒体再生增加和细胞氧化能量代谢改善。SIRT 1的作用及其在细胞水平上的作用机制尚不确定;然而，具有广泛研究焦点的研究人员有兴趣了解SIRT 1在不同组织中的生物学作用。为了研究这一点，需要产生条件性表达SIRT 1的转基因小鼠。为了响应NCRR的PA（PA-07-336），以开发适用于两个或多个分类NIH研究所/中心研究兴趣的人类疾病动物模型，我们将开发一种表达小鼠SIRT 1和四环素反应元件控制下的线粒体靶向增强型黄色荧光蛋白（TRE-SIRT 1/mito-eYFP）的转基因小鼠。将使用mito-eYFP与SIRT 1的共表达来鉴定、分离和研究SIRT 1表达对线粒体功能的影响。然后，SIRT 1表达将通过与CamKII-1 tTA小鼠杂交靶向中枢和外周神经元。本研究中开发的转基因小鼠将帮助NINDS，NIDDK，NIA和其他机构的研究人员测试疾病机制并开发SIRT 1介导的疗法。具体而言，在本提案中，我们将研究SIRT 1保护中枢和外周神经元免受糖尿病诱导的神经元损伤的机制。该提案还描述了其他研究人员如何开发所产生的动物模型，以研究非神经元细胞中的SIRT 1生物学。我们有两个目标：（1）建立四环素反应元件调控下表达小鼠SIRT 1的转基因小鼠（TRE; TRE-SIRT 1/mito-eYFP）。(2)对双基因SIRT 1神经元特异性小鼠模型进行表型分析。通俗描述：肥胖、糖尿病和神经退行性疾病影响着大量的人。SIRT 1蛋白被认为是身体防御疾病的主要调节剂，并被红葡萄中发现的白藜芦醇激活。我们的目标是揭示S1 RT 1保护神经元免受糖尿病引起的神经并发症的机制。 公共卫生关系：本提案是针对NCRR PA-07-336“动物模型和相关研究生物材料的开发”中的PA提交的。本PA的研究目标是“开发、表征或改进人类疾病的动物模型，并且所考虑的模型必须适用于两个或多个分类NIH研究所/中心的研究兴趣”。一个主要的公共卫生问题是与肥胖有关的疾病的发病率增加，例如糖尿病及其并发症，以及神经变性疾病的发病率增加，例如老年人中的帕金森病和阿尔茨海默病。我们将建立一个转基因小鼠，表达小鼠SIRT 1和四环素反应元件控制下的线粒体靶向增强型黄色荧光蛋白（TRE-SIRT 1/mito-eYFP）。将使用mito-eYFP与SIRT 1的共表达来鉴定、分离和研究SIRT 1表达对线粒体功能的影响。然后，SIRT 1表达将通过与CamKII-1 tTA小鼠杂交靶向中枢和外周神经元。本研究中开发的转基因小鼠将帮助NINDS，NIDDK，NIA和其他机构的研究人员测试疾病机制并开发SIRT 1介导的疗法。我们的总体假设是，SIRT 1在中枢（CNS）和外周神经系统（PNS）中的激活将减少氧化应激，并改善神经元和其他组织中Mt功能的调节，这在神经退行性疾病、糖尿病及其并发症中可能是重要的，并且在延迟或减少神经系统中的衰老效应中可能是重要的。sirtuins的激活为与氧化损伤和线粒体功能缺陷相关的几种人类疾病的新治疗提供了潜力。