AMPK localization, expression, and activity in Alzheimer's Disease

项目总结。阿尔茨海默病(AD)是痴呆症最常见的原因，据估计 65岁及以上的美国人中有十分之一患有阿尔茨海默病。美国阿尔茨海默氏症患者的数量将会上升 随着美国65岁及以上人口的规模和比例的增加，未来几年将迅速增长。没有 随着140多项临床试验的失败，AD已成为主要的疾病改变或治疗药物 公共卫生问题。人们一致认为，对这一疾病的病理生理学有更深入的了解 需要毁灭性的疾病才能推动这一领域向前发展。淀粉样蛋白-级联假说一直是领先的 近三十年来指导我们认识阿尔茨海默病病理生理的理论建构。然而， 这一结构忽略了最终导致神经退行性变和 该病的后续临床表现。已有假说认为异常代谢信号 是推动阿尔茨海默病的主要细胞机制。这些新陈代谢紊乱可能是由于 腺苷一磷酸活化蛋白激酶(AMPK)。AMPK是细胞能量的主要调节者 状态。尽管AMPK与AD有很强的联系，但它在AD中的特征还不完全：尚不清楚是如何发生的 亚细胞定位、活性和表达在人类阿尔茨海默病脑的脆弱区域发生了变化。 临床前文献表明，AMPK参与调节突触功能、A代谢、tau磷酸化、 阿尔茨海默病的病理性蛋白平衡。AMPK是由催化型()和调节型组成的必需的杂三聚体 (和)亚基。虽然亚单位在AD模型中的功能特征已经确定，但它的作用 受调控的亚基是未知的。因此，对人类底物中这种激酶的功能理解是 失踪。这一提议的中心假设是核AMPK活性和局部化在 广告。我们将使用死后大脑背外侧前额叶皮质和海马区来验证这一假说。 以认知障碍和AD患者为研究对象，分析AMPK随疾病进展的变化(具体目标1)。 同时，我们将使用一个组学平台来专门补充我们的生化研究。此外，我们还将 使用AD患者IPSC来源的皮质神经元来评估被忽视的调节AMPK1的作用 调节代谢和突触表型的亚单位(特定目标2)。这项提案针对的是一个关键的 需要解决AMPK在这种疾病的病理生理学中的作用。这项研究，与 经验丰富的指导团队，将为这位未来的研究员提供极好的培训经验。具体来说， 这项F30奖学金将支持申请者获得细胞生物学、生物信息学、干细胞培养、 基因操作策略和死后组织处理，为翻译生涯做准备 医生兼科学家。

PROJECT SUMMARY. Alzheimer’s disease (AD) is the most common cause of dementia and it is estimated that one in ten Americans aged 65 and older has AD. The number of Americans with Alzheimer’s will escalate rapidly in coming years as the size and proportion of the U.S. population aged 65 and older increases. With no disease-altering or curative drugs available, and with more than 140 failed clinical trials, AD has become a major public health concern. There is a consensus that a deeper understanding of the pathophysiology of this devastating illness is needed to move the field forward. The amyloid-cascade hypothesis has been the leading theoretical construct guiding our understanding of the pathophysiology of AD for the past thirty years. However, this construct has overlooked the multifaceted cellular mechanisms that ultimately drive neurodegeneration and the subsequent clinical manifestations of the disease. It has been hypothesized that aberrant metabolic signaling is a major cellular mechanism driving AD. These metabolic perturbations may arise through the dysfunction of adenosine monophosphate activated protein kinase (AMPK). AMPK is the master regulator of cellular energy status. Despite its strong association with AD, AMPK has not been fully characterized in AD: it is unknown how the subcellular localization, activity, and expression are changed in vulnerable regions of the human AD brain. Pre-clinical literature implicates AMPK in the regulation of synaptic function, A metabolism, tau phosphorylation, and pathologic proteostasis in AD. AMPK is an obligatory heterotrimer composed of catalytic () and regulatory ( and ) subunits. While the  subunit has been functionally characterized in AD models, the role of the regulatory subunits is unknown. As such, a functional understanding of this kinase in human substrates is missing. The central hypothesis of this proposal is that nuclear AMPK activity and localization is decreased in AD. We will test this hypothesis by using postmortem dorsolateral prefrontal cortex and hippocampus from mild cognitive impairment and AD subjects to analyze changes of AMPK as the disease progresses (Specific Aim 1). In parallel, we will employ an omics platform to specifically complement our biochemical studies. Further, we will use cortical neurons derived from AD-patient iPSCs to evaluate the role of the overlooked regulatory AMPK1 subunit in modulating a metabolic and synaptic phenotype (Specific Aim 2). This proposal addresses a critical need to resolve the role of AMPK in the pathophysiology of this disorder. This research, in conjunction with the experienced mentoring team, will provide this prospective Fellow an excellent training experience. Specifically, this F30 Fellowship will support the applicant in gaining expertise in cell biology, bioinformatics, stem cell culture, gene-manipulation strategies, and postmortem tissue processing, in preparation for a career as a translational physician-scientist.