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）：先前已经产生了p66 shc缺陷的小鼠，并且显示在一种特定的品系背景下具有延长的寿命（Migliaccio等人，1999年）。由于p66 shc缺陷是为数不多的几种遗传干预措施之一，据报道，增加哺乳动物的寿命，研究了解这一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产生缺乏p66 shc的小鼠。该动物模型是通过选择性地删除胚胎干（ES）细胞中p66 shc基因的外显子2的区域而产生的。我们建议使用新产生的p66 shc小鼠进行详细的研究p66 shc在衰老过程中的作用。待检验的中心假设是p66 shc缺陷将延缓衰老过程，并且这种作用是由于在靶组织中衰老期间ROS水平降低、线粒体功能的维持和ROS诱导的细胞凋亡减少。以骨骼肌为模型，我们将测试p66 shc缺陷是否会阻止与年龄相关的线粒体功能障碍、细胞凋亡和与年龄相关的转录改变。此外，我们建议确定p66 shc突变对长寿小鼠遗传背景中的寿命和疾病模式的影响。以前的研究提供了初步的实验支持p66 shc缺陷和延缓衰老过程之间的关联。特别是，p66 shc缺陷小鼠在129 Sv遗传背景下的寿命延长了约30%（平均寿命）。由于p66 shc是哺乳动物中唯一报道的突变，可能能够延长寿命的情况下，多种内分泌干扰，缺乏p66 shc的小鼠的详细表征是必要的。 p66 shc缺陷动物是本申请的焦点，应该允许详细表征p66 shc在长寿中的特定作用、其与ROS介导的衰老表型的关系以及p66 shc在骨骼肌衰老中的作用的特定检查。我们希望这种动物模型将提供一个非常有价值的工具，研究氧化应激和细胞凋亡在衰老中的作用。将在三个水平上评价衰老率：1）通过生存和疾病模式的生物体水平; 2）通过测量骨骼肌中的线粒体功能、氧化应激和细胞凋亡的生化和细胞水平;以及3）通过骨骼肌中的基因表达谱的转录水平。