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: 10359194
• 负责人: ANDREW N HOOFNAGLE
• 金额: $ 57.8万
• 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/10359194
• 关键词: Alternative Splicing; Alzheimer' s Disease; Alzheimer' s disease diagnosis; Alzheimer' s disease related dementia; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Behavior; Biochemical; Biological; Biological Assay; Biological Markers; Cause of Death; Cell Death; Cell Separation; 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; Institutes; Laboratories; Lead; Lipoproteins; Liquid Chromatography; Liquid substance; Mediator of activation protein; Methods; Molecular Conformation; Molecular Weight; Monitor; Neuraxis; 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.

摘要 阿尔茨海默氏病的病理特征是神经元缠结和淀粉样斑块， 在大脑皮质的灰质区域。错误折叠和聚集的蛋白质存在于那些 病理学特征，可能是神经元死亡的中心，导致局部萎缩，痴呆， 最终导致死亡浸泡中枢神经系统的脑脊髓液是最近端的液体 30多年来，它一直是生物标志物发现的焦点。有两个重要 存在于脑脊液中的基因产物，已被证明可预测疾病活动 和存在轻度认知损害的患者的认知下降：（1）淀粉样前体蛋白， 其产生Aβ（1-42）和其它片段，和（2）tau，其具有许多磷酸化位点。的 Aβ（1-42）作为淀粉样蛋白聚集体的沉积似乎导致邻近神经元的死亡。所得 损伤增加了释放到脑脊髓液中的tau蛋白的量。来自既往生物标志物 根据对Aβ蛋白和tau蛋白的研究，似乎整个疾病过程，从Aβ（1-42）沉积到细胞 死亡，与其他神经退行性疾病不同。因此，这是一个引人注目的 假设脑脊液中存在可能是疾病介质的新生物标志物 并且可以在目前用于诊断阿尔茨海默病的生物标志物上添加更多信息。 先前已经尝试使用发现蛋白质组学来鉴定差异表达的蛋白质。 与对照组相比，阿尔茨海默病患者的脑脊液中有 很少有人努力以临床有意义的方式扩展这些研究。然而，其中一些努力 已经鉴定了淀粉样前体蛋白的另外片段的存在[即，除Aβ（1-42）外] 和其他蛋白质，这表明，翻译后的变化可能是重要的形成， 淀粉样蛋白、神经元突触功能障碍和随后的神经元死亡。 在这个项目中，题为“鉴定质谱目标和发展多重测定， 疾病管理，”我们的目标是确定翻译后修饰，导致改变 脑脊液中LP/EV浓度、分子量和稳定性。部分原因是， Aβ（1-42）、tau蛋白和磷酸化tau蛋白作为脑脊液生物标志物的临床翻译花了几十年时间 早期没有可重复的检测方法。因此，该项目的另一个首要目标是 是开发精确的，可转移的，经过验证的靶向蛋白质组学检测，以定量蛋白质， 脑脊液，可用于研究疾病机制和预测不良结果。