Cellular mechanisms underlying age-associated changes in sleep and oxidative homeostasis

项目摘要 在整个动物界，睡眠是一种进化上保守的行为。在人类和模型生物中， 睡眠不足会导致疾病甚至死亡。此外，在人类和模式生物中，衰老都会导致 睡眠模式的改变和睡眠质量的下降。睡眠质量下降也与许多 与年龄相关的疾病，如人类的阿尔茨海默氏症。因此，睡眠被认为起着至关重要的作用 在健康衰老中的作用。与年龄相关的疾病也与氧化应激的生物标记物增加有关， 活性氧(ROS)升高和细胞氧化损伤的状态。使用黑腹果蝇， Shirasu-Hiza实验室之前曾表明，睡眠有助于抵御氧化应激。根据这些数据， 我的中心假设是，睡眠在特定的组织中起作用，激活细胞氧化应激反应通路 而且，随着睡眠质量随着年龄的增长而下降，这些氧化应激防御系统会减弱，并允许与年龄相关的 病理生理学。由于潜在的机制尚不清楚，我建议将睡眠诱导 防御氧化应激的细胞机制并确定这些机制在正常衰老过程中如何变化 在阿尔茨海默病模型中也是如此。我将使用黑腹果蝇，一种有利的模式生物 这是因为睡眠和氧化应激的潜在机制从苍蝇到人类都是保守的 而且已经建立了阿尔茨海默病的果蝇模型。目标1将识别特定组织(S)，其中 睡眠促进了对氧化应激的防御，并决定了这些是否随着年龄的增长而变得更脆弱。ROS 将评估睡眠较短的幼蝇的组织中的水平和氧化损伤，相对于对照组和 与中老年苍蝇相比。目标2将研究睡眠质量如何调节氧化应激状态 以及阿尔茨海默病模型的表型。ROS水平、氧化损伤与急性脑损伤后的存活率 氧化应激将被用来评估阿尔茨海默病模型果蝇的氧化应激状态 睡眠不足相对于未经操控的阿尔茨海默病苍蝇和对照。疾病的严重程度将被评估 通过寿命、流动性和蛋白质聚集。目标3将研究特定的细胞机制，通过 睡眠促进了对氧化应激的防御，以及这些防御是如何随着年龄的变化而变化的。RNA测序将是 用来研究短眠果蝇和对照组在常氧和低氧条件下的转录差异 高氧、高氧治疗。显著差异表达的基因和/或途径将被评估 在中老年果蝇睡眠促进的氧化应激防御中的功能作用 作为阿尔茨海默氏症模型。总之，这些实验将确定睡眠和睡眠之间的细胞联系 氧化应激防御以及这种关系如何随年龄和阿尔茨海默病的病理变化而变化。这 将提高我们对衰老、阿尔茨海默氏症和睡眠治疗潜力的理解。

Project Summary Sleep is an evolutionarily conserved behavior across the animal kingdom. In both humans and model organisms, a lack of sleep leads to illness and even death. Moreover, in both humans and model organisms, aging leads to changes in sleep patterns and declines in sleep quality. Declining sleep quality is also associated with many age-related diseases, such as Alzheimer’s disease in humans. Therefore, sleep is thought to play an essential role in healthy aging. Age-related diseases are also associated with increased biomarkers of oxidative stress, a state of elevated reactive oxygen species (ROS) and cellular oxidative damage. Using Drosophila melanogaster, the Shirasu-Hiza lab previously showed that sleep promotes defense against oxidative stress. Given these data, my central hypothesis is that sleep acts in specific tissues to activate cellular oxidative stress response pathways and that, as sleep quality declines with age, these oxidative stress defenses weaken and allow age-related pathophysiologies. Because the underlying mechanisms remain unclear, I propose to identify sleep-induced cellular mechanisms that defend against oxidative stress and determine how these change with normal aging and in Alzheimer’s disease models. I will use Drosophila melanogaster, an advantageous model organism for this work because mechanisms underlying both sleep and oxidative stress are conserved from flies to humans and there are established Drosophila models of Alzheimer’s disease. Aim 1 will identify specific tissue(s) in which sleep promotes defense against oxidative stress and determine if these are more vulnerable with age. ROS levels and oxidative damage will be assessed in tissues of young short-sleeping flies relative to controls and compared to middle-aged and old flies. Aim 2 will examine how sleep quality modulates the oxidative stress state and phenotypes of Alzheimer’s disease models. Levels of ROS, oxidative damage, and survival after acute oxidative stress will be used to assess the oxidative stress state of Alzheimer’s model flies with induced or deprived sleep relative to unmanipulated Alzheimer’s disease flies and control. Disease severity will be assessed through lifespan, mobility, and protein aggregation. Aim 3 will investigate specific cellular mechanisms by which sleep promotes defense against oxidative stress and how these change with age. RNA-sequencing will be employed to probe the transcriptional differences between short-sleeping flies and controls under normoxia and hyperoxia, high oxygen treatment. Significantly differentially expressed genes and/or pathways will be assessed for their functional role in sleep-promoted defense against oxidative stress in middle-aged and old flies, as well as Alzheimer’s models. Together, these experiments will determine the cellular connection between sleep and oxidative stress defense and how this relationship changes with age and Alzheimer’s disease pathology. This will improve our understanding of aging, Alzheimer’s disease, and the therapeutic potential of sleep.