A Novel p16INK4a Reporter System to Assess Aging In Vivo

6.项目摘要/摘要： 随着美国人口的增加，与年龄有关的疾病的发病率继续上升 65岁以上的人会增长。因此，研究现在集中在发现方法上。 以延年益寿。尽管生物年龄的分子指标很少 经过鉴定，我们和其他人最近已经表明，p16INK4a具有强大的 哺乳动物衰老的准确标志。P16INK4a的诱导与 啮齿动物的时间年龄，可以通过卡路里等抗衰老策略来延缓 限制。此外，我们的初步数据显示，p16INK4a水平下降了 健康受试者的适度运动。尽管有这些数据，但这些机制 改变哪种生活方式能延长寿命在很大程度上还没有定论。在这里，我会 利用p16INK4a的独特性质作为老化生物标记物来评估组织- 抗衰老策略的特殊性。为了做到这一点，我开发了一个新颖的敲门记者 表达萤火虫荧光素酶的等位基因p16-Luc P16INK4a启动子。重要的是，我的初步数据显示，该等位基因的诱导 在体外与细胞衰老相关，并可在活体动物中观察到。 利用这些小鼠，我计划解决以下具体目标：1)表征p16- LUC等位基因在与哺乳动物衰老过程相关的条件下，2)调查 饮食和运动对p16INK4a诱导的影响，3)确定诱导的时间。 饮食和运动对p16INK4a启动子的依赖性和器官特异性影响 入住率。

6. Project Summary/Abstract: The incidence of age-related diseases continues to rise as the number of Americans over the age of 65 grows. As a result, research is now focused on discovering methods to promote longevity. Although few molecular indicators of biological age have been indentified, we and others have recently shown that p16INK4a serves a robust and accurate marker of mammalian aging. Induction of p16INK4a correlates directly with chronological age in rodents and can be slowed by anti-aging strategies such as caloric restriction. In addition, our preliminary data show that p16INK4a levels are decreased by moderate exercise in healthy human subjects. Despite these data, the mechanisms by which lifestyle modification promotes longevity remain largely undefined. Herein, I will exploit the unique properties of p16INK4a as an aging biomarker to assess the tissue- specificity of anti-aging strategies. To do this, I have developed a novel knockin reporter allele, p16-LUC, which expresses firefly luciferase under control of the endogenous p16INK4a promoter. Importantly, my preliminary data show that induction of this allele correlates with cellular senescence in vitro and can be visualized in living animals. Using these mice I plan to address the following specific aims: 1) Characterize the p16- LUC allele under conditions relevant to the mammalian aging process, 2) Investigate the influence of diet and exercise on the induction of p16INK4a, 3) Determine the time- dependent and organ-specific effects of diet and exercise on p16INK4a promoter occupancy.