Novel multiplexing proteomics to study the periphery in Alzheimer's disease

 描述（由申请人提供）：阿尔茨海默病（AD）无法治愈。全国有500万人患有AD，随着婴儿潮一代的继续老龄化，这一数字将增长到2000万。即使有关于AD的当前临床和病理学知识，随着疾病进展发生的确切原因和因素也没有完全理解。有趣的是，肝脏产生有毒的淀粉样β肽，穿过血脑屏障。用药物减少肝脏中这些肽的产生可以降低小鼠大脑中淀粉样β肽的水平。关于肝脏和中枢神经系统（CNS）以外的其他器官的变化如何影响AD的发作、进展和治疗仍存在问题。我们的实验室已经表明，在AD小鼠模型的晚期阶段，肝脏和外周T细胞中的脂质和能量代谢以及氧化应激途径发生了改变。这些变化可能与已知在CNS中发生的这些通路的类似动态平衡障碍有关。为了更好地了解AD，有必要研究CNS以外的变化，并充分了解它们与CNS的关系。PI的长期目标是提供更好的信息，分子变化，引发疾病发作的AD和使用这些信息，以帮助促进新的途径，AD治疗。定量蛋白质组学是解决这一问题的有力工具，因为它告诉我们特定细胞功能的变化。不过，目前还无法多路复用 到回答关于外周器官在AD中的作用的问题所必需的程度。在这里，我们提出了三个目标来解决这个问题：目标1。开发多路复用功能，在前所未有的60个组织样本中定量测量相对蛋白质表达。目标2.创建选择性多重蛋白质组学，用于含半胱氨酸蛋白质的翻译后修饰，以了解氧化应激对组织中蛋白质的影响。目标3.首次阐明AD小鼠和年龄/性别匹配对照组在疾病症状发作前后的外周组织和多个外周器官（肝、心、肾、肺和脑）中的AD发病机制。这些蛋白质组学测量将确定外周代谢和氧化应激途径的改变如何与CNS的变化相关。此外，将确定这些变化与疾病发作和进展的时间。这些目标的成功完成将有助于阐明外周器官如何促进AD病理学，并有助于促进AD治疗的新途径。这一结果对于促进NIH的使命“进行人类疾病的病因、诊断、预防和治疗的研究”是必要的。

 DESCRIPTION (provided by applicant): There is no cure for Alzheimer's Disease (AD). Five million people nationwide are suffering from AD and this number will grow to 20 million as the baby boomer generation continues to age. Even with current clinical and pathological knowledge about AD, the exact causes and factors that take place with disease progression are not fully understood. Interestingly, the liver generates toxic amyloid-β peptides that travel acros the blood brain barrier. Decreasing the liver's production of these peptides with drugs decreases brain amyloid-beta peptide levels in mice. Questions remain concerning how changes in the liver and other organs outside the central nervous system (CNS) influence onset, progression, and treatment of AD. Our laboratory has already shown that lipid and energy metabolism as well as oxidative stress pathways are altered in liver and peripheral T-cells in advanced stages of an AD mouse model. These changes may be connected to similar dyshomeostasis of these pathways that are well known to occur in the CNS. In order to gain a better understanding of AD, it is necessary to study changes outside of the CNS and fully understand how they relate to CNS. The PI's long-term objectives are to provide better information about molecular changes that initiate disease onset in AD and use this information to help facilitate new avenues for AD therapeutics. Quantitative proteomics is a powerful tool to apply to this problem because it tells us about changes in specific cellular functions. However, it is currently not possible to multiplex to the degree necessary for answering questions about the role of peripheral organs in AD. Here, we propose three aims to address this issue: Aim 1. Develop multiplexing capabilities to measure relative protein expression quantitatively in an unprecedented 60 tissue samples. Aim 2. Create selective multiplexing proteomics for post-translational modification of cysteine- containing proteins to understand the impact of oxidative stress on proteins in tissue. Aim 3. Elucidate AD pathogenesis in peripheral tissues for the first time to multiple peripheral organs (liver, heart, kidney, lung, and brain) from AD mice and age/sex-matched controls at ages before and after the onset of disease symptoms. These proteomics measurements will establish how altered metabolic- and oxidative-stress pathways in the periphery relate to changes in the CNS. Furthermore, the timing of these changes with disease onset and progression will be established. Successful completion of these aims will help to delineate how peripheral organs contribute to AD pathology and help facilitate new avenues for AD therapeutics. This outcome is necessary to further the mission of the NIH "to conduct research in the causes, diagnosis, prevention, and cure of human diseases".