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

项目摘要。阿尔茨海默病（AD）是痴呆症的最常见原因，据估计， 65岁以上的美国人中有十分之一患有AD。美国老年痴呆症患者的数量将逐步增加 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 目前已有140多项临床试验失败，AD已成为一种主要的 公共卫生问题。有一个共识是，更深入地了解这一病理生理学， 需要毁灭性的疾病来推动这一领域的发展。淀粉样蛋白级联假说一直是 在过去的30年里，这是一个指导我们理解AD病理生理学的理论框架。然而，在这方面， 这种结构忽略了最终驱动神经变性的多方面细胞机制， 疾病的后续临床表现。有人假设异常的代谢信号 是导致AD的主要细胞机制这些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因子 status.尽管AMPK与AD密切相关，但其在AD中的特征尚未完全确定： 亚细胞定位、活性和表达在人AD脑的易受伤害区域中发生改变。 临床前文献暗示AMPK参与调节突触功能、腺苷酸代谢、tau磷酸化， 和病理性蛋白质稳态。AMPK是一种强制性的异源三聚体，由催化性（AMPK）和调节性（AMPK）组成。 （α和β）亚基。虽然AD模型中的β亚基已被功能性地表征，但β亚基的作用仍有待进一步研究。 调节亚基是未知的。因此，对该激酶在人类底物中的功能理解是 失踪了这一建议的中心假设是，核AMPK活性和定位降低， AD.我们将通过使用轻度脑损伤患者的死后背外侧前额叶皮层和海马来验证这一假设。 认知障碍和AD受试者，以分析AMPK随疾病进展的变化（具体目标1）。 同时，我们将采用组学平台来专门补充我们的生化研究。此外，我们将 使用来自AD患者iPSC的皮质神经元来评估被忽视的调节AMPK β 1的作用。 亚基在调节代谢和突触表型中的作用（特异性目的2）。该提案涉及一个关键问题， 需要解决AMPK在这种疾病的病理生理学中的作用。这项研究，结合 经验丰富的指导团队，将提供这一未来的研究员一个很好的培训经验。具体地说， 该F30奖学金将支持申请人获得细胞生物学，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。 物理学家兼科学家

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.