Oxidative Stress, p66shc and the Aging Process

氧化应激、p66shc 和衰老过程

• 批准号: 6866933
• 负责人: TOMAS ALBERTO PROLLA
• 金额: $ 28.9万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6866933
• 关键词: aging; apoptosis; biological signal transduction; cytochrome oxidase; developmental genetics; electron transport; free radical oxygen; gene expression profiling; laboratory mouse; mitochondria; mitochondrial disease /disorder; oxidative stress; striated muscles

DESCRIPTION (provided by applicant): Mice deficient for p66shc have been generated previously and shown to have extended lifespan in one particular strain background (Migliaccio et al., 1999). Because p66shc deficiency is one of the few genetic interventions reported to increase lifespan in a mammal, studies to understand how this pathway impacts lifespan have the potential to increase our understanding of aging mechanisms. We have used gene targeting to generate mice that lack p66shc. This animal model was generated by selectively deleting a region of exon 2 of the p66shc gene in embryonic stem (ES) cells. We propose to use the newly generated p66shc mice to conduct a detailed study of the role of p66shc on the aging process. The central hypothesis to be tested is that p66shc deficiency will retard the aging process, and that this effect is due to reduced ROS levels, maintenance of mitochondrial function and reduced ROS-induced apoptosis during aging in target tissues. Using skeletal muscle as a model, we will test if p66shc deficiency will prevent age-related mitochondrial dysfunction, apoptosis, and age-related transcriptional alterations. Further, we propose to determine the effect of p66shc mutation on lifespan and disease patterns in a long-lived mouse genetic background. A previous study has provided preliminary experimental support for an association between p66shc deficiency and retardation of the aging process. In particular, p66shc deficient mice were shown to live approximately 30% longer (average lifespan) in a 129Sv genetic background. Because p66shc is the only reported mutation in mammals that may be able to extend lifespan in the absence of multiple endocrine disruption, a detailed characterization of mice lacking p66shc is needed. p66shc deficient animals, which are the focus of this application, should allow detailed characterization of the specific role of p66shc in longevity, its relationship to ROS mediated aging phenotypes, and a specific examination of the role of p66shc in skeletal muscle aging. We expect that this animal model will provide an extremely valuable tool to investigate the role of oxidative stress and apoptosis in aging. Aging rates will be evaluated at three levels: 1) organismal level by survival and disease patterns; 2) biochemical and cellular level by measures of mitochondrial function, oxidative stress and apoptosis in skeletal muscle; and, 3) transcriptional level, through gene expression profiling in skeletal muscle.

描述（由申请人提供）：先前已经生成了缺乏P66SHC的小鼠，并在一个特定的菌株背景下具有延长的寿命（Migliaccio等，1999）。由于p66SHC缺乏症是据报道会在哺乳动物中增加寿命的少数几个遗传干预措施之一，因此了解该途径如何影响寿命有可能增加我们对衰老机制的理解。我们已经使用基因靶向来产生缺乏P66SHC的小鼠。该动物模型是通过选择性地删除胚胎茎（ES）细胞中P66SHC基因2的外显子2的区域而产生的。我们建议使用新生成的P66SHC小鼠对P66SHC在衰老过程中的作用进行详细研究。要测试的中心假设是p66SHC缺乏症会阻碍衰老过程，并且这种作用是由于ROS水平降低，线粒体功能的维持以及靶组织衰老期​​间ROS诱导的凋亡降低所致。使用骨骼肌作为模型，我们将测试P66SHC缺乏症是否会防止与年龄有关的线粒体功能障碍，凋亡和与年龄相关的转录改变。此外，我们建议在长寿命小鼠遗传背景下确定p66SHC突变对寿命和疾病模式的影响。先前的研究为P66SHC缺乏症与衰老过程的延迟之间的关联提供了初步的实验支持。特别是，在129SV遗传背景中，p66SHC缺乏小鼠的寿命约为30％（平均寿命）。由于p66SHC是哺乳动物中唯一报告的突变，在没有多种内分泌干扰的情况下可能能够延长寿命，因此需要对缺乏P66SHC的小鼠的详细表征。 P66SHC缺乏动物，这是本应用的重点，应允许详细描述p66SHC在寿命中的特定作用，其与ROS介导的衰老表型的关系以及对p66SHC在骨骼肌衰老中的作用的特定检查。我们预计该动物模型将为研究氧化应激和凋亡在衰老中的作用提供极为有价值的工具。衰老率将在三个级别评估：1）生存和疾病模式的生​​物水平； 2）通过测量线粒体功能，氧化应激和骨骼肌凋亡的量度； 3）转录水平，通过骨骼肌中的基因表达分析。