Determining the role of OXR1 in aging and Alzheimer's disease

项目摘要/摘要 尽管确定了一些阿尔茨海默病的遗传风险因素，但大多数晚发病例的确切病因 阿尔茨海默病(LOAD)是未知的。随后，治疗阿尔茨海默病的疗法基本上都没有成功。 衰老和饮食是AD的两个重要危险因素，其中衰老是AD的最大危险因素，在很大程度上仍然存在 已被忽略。饮食限制(DR)是延缓衰老的最有力的干预措施之一，也延缓了 阿尔茨海默病(AD)跨物种的多种模型。我们利用了短暂的寿命和强大的 利用遗传工具鉴定芥菜(MTD)/氧化应激抵抗蛋白1(OXR1) DR对寿命的保护作用需要在神经元中进行。重要的是，我们已经观察到OXR1 苍蝇和人诱导的多能干细胞(IPSC)对衰老相关神经变性的保护作用 神经退行性疾病的模型。OXR1保护神经退行性变的机制 目前仍不清楚。我们观察到，抑制OXR1减少了逆转聚体蛋白，而增强了逆转聚体 功能，挽救抑制OXR1的有害影响。此外，我们发现OXR1和OXR1的改变 几种逆转聚体蛋白与人类AD风险增加有关，使用来自 来自加速药物计划-阿尔茨海默病(AMP-AD)网络的1000多名AD患者。这里, 我们建议检验OXR1增强逆转录功能以延缓衰老和神经退变的假设 采用苍蝇和人IPSC AD模型。在第一个目标中，我们将确定监管机制 OXR1以及它如何影响衰老和与衰老相关的神经元损伤。在第二个目标中，我们将确定 OXR1在增强逆转录因子功能中的DR依赖作用及逆转录因子在介导血管紧张素转换酶中的作用 DR的保护作用：确定OXR1增强DR逆转录功能的机制 将使用蛋白质组学来确定和表征OXR1的蛋白质结合伙伴。在第三个目标中，我们将 在AD模型中检测OXR1在防止神经退行性变中的作用，并通过增强逆转录作用 功能。我们将在果蝇模型中测试OXR1是否调节AD病理，该模型过度表达人tau或 淀粉样蛋白β(Aβ)。我们将在前脑胆碱能神经元和皮质神经元中过表达OXR1和逆转录病毒蛋白 并测量AD终点：β42/40蓄积、细胞死亡和电生理学。 因为OXR1在果蝇中调节逆转录功能，所以我们将评估这一调节是否在 人AD来源的IPSCs和进行组学方法以确定介导OXR1OXR‘S的关键信号通路 起到保护作用。通过表征OXR1调节的逆转录功能和蛋白质网络及其在 衰老和年龄依赖性神经退变，我们将提供新的靶点，开发治疗方法来减缓 与AD相关的病理和延长健康寿命。此外，我们将确定是否重复使用蛋白质 通过营养限制条件下的逆转录可以起到神经保护作用，延缓衰老。

PROJECT SUMMARY/ABSTRACT Despite identifying some of the genetic risk factors for AD, the precise etiology of most cases of Late-Onset Alzheimer's disease (LOAD) is unknown. Subsequently, therapies to treat AD have been largely unsuccessful. Two important risk factors for AD, aging, and diet, with aging being the greatest risk factor for AD, remain largely ignored. Dietary restriction (DR), one of the most robust interventions to slow aging, also delays the onset of Alzheimer’s disease (AD) in multiple models across species. We exploited the short lifespan and powerful genetic tools in D. melanogaster to identify that mustard (mtd)/oxidative stress resistance protein 1 (OXR1) in neurons is required for the protective effects of DR on lifespan. Importantly, we have observed that OXR1 protects against age-related neurodegeneration in fly and human-induced pluripotent stem cell (iPSC) derived models of neurodegenerative diseases. The mechanisms by which OXR1 protects against neurodegeneration remains unclear. We observed that inhibiting OXR1 reduces retromer proteins while enhancing retromer function, rescues the deleterious effects of inhibiting OXR1. Furthermore, we found that alterations in OXR1 and several retromer proteins are associated with an increased risk of AD in humans using proteomics data from over 1000 AD patients from the Accelerating Medicines Program-Alzheimer’s Disease (AMP-AD) network. Here, we propose to test the hypothesis that OXR1 enhances retromer function to slow aging and neurodegeneration using fly and human iPSC models of AD. In the first aim, we will determine the mechanisms of regulation of OXR1 and how that influences aging and age-related neuronal damage. In the second aim, we will determine the DR-dependent role of OXR1 in enhancing retromer function and the role of retromer in mediating the protective effects of DR. To determine the mechanism by which OXR1 enhances retromer function upon DR; we will use proteomics to determine and characterize the protein binding partners of OXR1. In the third aim, we will test the role of OXR1 in protecting against neurodegeneration in models of AD, and by enhancing retromer function. We will test whether OXR1 modulates AD pathology in fly models that overexpress human tau or amyloid β (Aβ). We will overexpress OXR1 and retromer proteins in forebrain cholinergic and cortical neuron derived AD iPSCs and measure AD endpoints: Aβ42/40 accumulation, cell death, and electrophysiology. Because OXR1 regulates retromer function in the fly, we will evaluate whether this regulation is conserved in human AD-derived iPSCs and carry out omics approaches to identify key signaling pathways mediating OXR1’s protective effects. By characterizing retromer function and protein networks regulated by OXR1 and their role in aging and age-dependent neurodegeneration, we will provide novel targets for developing therapeutics to slow AD-related pathologies and extend healthspan. Furthermore, we will determine whether reuse of proteins through retromer under nutrient limiting conditions is neuroprotective and slows aging.