Project 3: Development of multiplex assays for clinical monitoring of disease

项目 3：开发用于疾病临床监测的多重检测方法

• 批准号: 10573266
• 负责人: ANDREW N HOOFNAGLE
• 金额: $ 60.82万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10573266
• 关键词: Alternative Splicing; Alzheimer' s Disease; Alzheimer' s disease diagnosis; Alzheimer' s disease patient; Alzheimer' s disease related dementia; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Bathing; Behavior; Biochemical; Biological; Biological Assay; Biological Markers; Cause of Death; Cell Death; Cell Separation; Central Nervous System; Cerebral cortex; Cerebrospinal Fluid; Cessation of life; Clinical; Computer software; Data; Dementia; Deposition; Development; Disease; Disease Management; Ensure; Event; Goals; Guidelines; Hand; Impaired cognition; Injury; Laboratories; Lipoproteins; Liquid Chromatography; Liquid substance; Mediator; Methods; Molecular Conformation; Molecular Weight; Monitor; Neurodegenerative Disorders; Neurofibrillary Tangles; Neuronal Dysfunction; Neurons; Outcome; Pathologic; Patients; Phosphorylation Site; Post-Translational Protein Processing; Procedures; Process; Proteins; Proteomics; Reproducibility; Research Personnel; Sampling; Senile Plaques; Speed; Synapses; Tissue imaging; Validation; abeta deposition; amyloid formation; biomarker discovery; brain tissue; clinical translation; differential expression; experience; extracellular vesicles; gene product; gray matter; interest; mild cognitive impairment; multiplex assay; neuron loss; next generation; novel; novel marker; protein aggregation; regional atrophy; success; tandem mass spectrometry; tau Proteins; tau-1; translational proteomics; verification and validation

Abstract The pathological hallmarks of Alzheimer’s disease are the neurofibrillary tangles and amyloid plaques that form in gray matter regions of the cerebral cortex. Misfolded and aggregated proteins reside within those pathological features, which may to be central to the neuronal death that causes regional atrophy, dementia, and ultimately death. The cerebrospinal fluid that bathes the central nervous system is the fluid most proximal to the disease and has been the focus of biomarker discovery for more than 30 years. There are two important gene products that are present in cerebrospinal fluid and have been shown to be predictive of disease activity and cognitive decline in patients that present with mild cognitive impairment: (1) the amyloid precursor protein, which gives rise to Aβ(1-42) and other fragments, and (2) tau, which has many phosphorylation sites. The deposition of Aβ(1-42) as amyloid aggregates appears to lead to the death of adjacent neurons. The resulting injury increases the amount of tau protein released into the cerebrospinal fluid. From previous biomarker studies of Aβ proteins and tau, it seems likely that the entire disease process, from Aβ(1-42) deposition to cell death, is orchestrated differently than in other neurodegenerative diseases. As a result, it is a compelling hypothesis that novel biomarkers, which may be mediators of disease, are present in the cerebrospinal fluid and could add more information over the current biomarkers used in the diagnosis of Alzheimer’s disease. There have been previous attempts to use discovery proteomics to identify proteins that are differentially expressed in cerebrospinal fluid in patients with Alzheimer’s disease compared with controls and there have been few efforts to expand upon these studies in a clinically meaningful way. However, some of these efforts have identified the presence of additional fragments of amyloid precursor protein [i.e., in addition to Aβ(1-42)] and other proteins, which suggests that post-translational changes may be important in the formation of amyloid, the dysfunction of neuronal synapses, and the subsequent death of neurons. In this project entitled, “Identification of mass spectrometric targets and development of multiplex assays for disease management,” our objective is to identify the post-translational modifications that lead to altered LP/EV concentration, molecular weight, and stability in cerebrospinal fluid. Part of the reason that the clinical translation of Aβ(1-42), tau, and phosphorylated tau as cerebrospinal fluid biomarkers took decades was that reproducible assays were not available early on. As a result, another overarching goal of this project is to develop precise, transferable, validated targeted proteomic assays to quantify proteins in cerebrospinal fluid that can be used to investigate disease mechanism and predict poor outcomes.

抽象的 阿尔茨海默氏病的病理标志是形成的神经原纤维缠结和淀粉样斑块 在大脑皮层的灰质区域。错误折叠和聚集的蛋白质存在于其中 病理特征，这可能是导致区域萎缩，痴呆的神经元死亡的核心 并最终死亡。沐浴中枢神经系统的脑脊液是最近端的液体 对于该疾病，已经是生物标志物发现的重点已有30多年了。有两个重要 脑脊液中存在的基因产物，已被证明是疾病活性的 出现轻度认知障碍的患者的认知能力下降：（1）淀粉样蛋白前体蛋白， 产生Aβ（1-42）和其他片段，以及（2）Tau，它具有许多磷酸化位点。 Aβ（1-42）作为淀粉样蛋白聚集体的沉积似乎导致相邻神经元死亡。结果 损伤增加了释放到脑脊液中的tau蛋白的量。来自以前的生物标志物 对Aβ蛋白和TAU的研究，从Aβ（1-42）沉积到细胞的整个疾病过程似乎很可能 死亡与其他神经退行性疾病的策划不同。结果，这是一个引人注目的 假设新型生物标志物可能是疾病的介体，存在于脑脊液中 并且可以添加有关当前用于诊断阿尔茨海默氏病的生物标志物的信息。 以前曾尝试使用发现蛋白质组学来识别差异化的蛋白质 与对照组相比，在阿尔茨海默氏病患者的脑脊液中表达 我们很少努力以临床意义的方式扩展这些研究。但是，其中一些努力 已经确定存在淀粉样前体蛋白的其他片段[即除了Aβ（1-42）] 和其他蛋白质，这表明翻译后变化在形成中可能很重要 淀粉样蛋白，神经元突触的功能障碍以及随后的神经元死亡。 在这个项目中，题为“质谱目标的识别和开发多重测定法 疾病管理，“我们的目标是确定导致改变的翻译后修饰 脑脊液中的LP/EV浓度，分子量和稳定性。部分原因 Aβ（1-42），tau和磷酸化的tau作为脑脊液生物标志物的临床翻译花费数十年 是可再现的阿萨斯是不早于的。结果，该项目的另一个总体目标 是为了开发精确，转移，经过验证的靶向蛋白质组学测定，以量化 脑脊液可用于研究疾病机制并预测不良预后。