Linkage-specific ubiquitylation patterns as highly sensitive markers for neurodegenerative disease

连锁特异性泛素化模式作为神经退行性疾病的高度敏感标记

• 批准号: 9789790
• 负责人: Tauseef R. Butt
• 金额: $ 77.97万
• 依托单位: LIFESENSORS, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9789790
• 关键词: Affinity; Age; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Autophagosome; Binding; Biological Assay; Biological Markers; Blood Tests; Cell Line; Cell physiology; Cells; Cerebrospinal Fluid; Clinic; Clinical Trials; Complex; DNA Repair; Databases; Detection; Development; Diagnosis; Disease; Disease Progression; Drug Targeting; Engineering; Enzymes; Etiology; Event; FDA approved; Failure; Functional disorder; Goals; Grant; Hyperactive behavior; Imaging Device; Individual; Lysine; Lysosomes; Mass Spectrum Analysis; Mediating; Medicine; Methods; Molecular Profiling; Monitor; N-terminal; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurology; Neurons; Pathogenicity; Pathologic; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Pharmacology; Phase; Plant Roots; Play; Polymers; Polyubiquitin; Population; Positioning Attribute; Progress Reports; Proteins; Proteomics; Research; Ribosomes; Risk; Role; Sampling; Serum; Serum Proteins; Signal Transduction; Stable Isotope Labeling; Symptoms; System; Techniques; Technology; Testing; Translating; Ubiquitin; Universities; alpha synuclein; base; biomarker panel; brain tissue; clinical efficacy; commercialization; diagnostic assay; dosage; link protein; misfolded protein; multicatalytic endopeptidase complex; new therapeutic target; novel; patient population; patient stratification; programs; protein TDP-43; protein aggregate; protein biomarkers; protein transport; proteomic signature; proteostasis; response; success; targeted biomarker; tau Proteins; tool; treatment strategy

While sensitive imaging tools to detect β amyloid and tau proteins and other biomarkers for established Alzheimer’s disease (AD) and other neurodegenerative diseases are available, similar tools for detecting pre- symptomatic, etiologic biomarkers of these diseases are not. Among the earliest steps in AD is a failure of the ubiquitin proteasome pathway to respond to misfolded proteins by causing their degradation, thereby promoting aggregation. This early pathological event is marked in part by changes in protein ubiquitylation patterns in neuronal cells. “Ubiquitylation patterns” encompasses multiple ubiquitin chains and lysine positions, and this information is far more complex than a simple ubiquitylation event. In addition, ubiquitylation patterns in neuronal cells of AD patients change as the disease progresses. No non-invasive blood test for pre-symptoms or etiology of Alzheimer ’s disease has been approved by the FDA. In addition to diagnosis, the ability to identify molecular signatures of AD could help stratify the patient population, and changes in ubiquitylation patterns could be used to monitor the clinical efficacy of drugs that target the ubiquitin proteasome system. The aim of this project is to capture the initial step in neurodegenerative disease establishment as well as stages in disease progression by identifying appropriate ubiquitylation pattern signatures that can be detected routinely in a diagnostic assay. Toward this end, in Phase I, LifeSensors developed a highly sensitive method to isolate poly-ubiquitylated proteins based on Ub chain selectivity using TUBES – an affinity matrix that can bind selectively to various distinct poly-Ub chains -- in concert with mass spectrometry (MS) to identify molecular signatures based on ubiquitylation. Conditions were simulated in neuronal cells that mimic neurodegeneration, and marker proteins were identified; in addition, methods were developed to identify low levels of ubiquitylated proteins and observe changes in ubiquitylation patterns from cerebrospinal fluid (CSF) of normal and AD patients. In Phase II, it is proposed to extend these studies to examine CSF and serum ubiquitylated protein molecular signatures from approximately 100 normal and AD patients. LifeSensors will collaborate with Drs. Nicholas Seyfried and Allan Levey of the Alzheimer’s Disease Research Center, leaders in AD proteomics, to translate the tools developed in Phase I to monitor ubiquitylation pattern changes in CSF of normal controls and AD patients and to establish biomarker panels that distinguish controls from AD patients at various disease stages, validating the markers using existing databases and predictive studies. Success in Phase II will lead to the discovery of novel ubiquitylation signatures for various stages of AD. LifeSensors will commercialize stable isotope labeled versions of these proteins to quantify the markers by mass spec in the serum and/or CSF. These simple tests will stratify patient populations to monitor efficacy in clinical trials and guide the choice and dosage of drug to treat AD.

检测 β 淀粉样蛋白和 tau 蛋白以及其他生物标志物的灵敏成像工具已建立 阿尔茨海默氏病 (AD) 和其他神经退行性疾病已有类似的工具可用于检测预兆 这些疾病的症状、病因生物标志物并不是 AD 的最早步骤之一。 泛素蛋白酶体途径通过引起错误折叠的蛋白质降解来对其做出反应，从而促进 这种早期病理事件的部分特征是蛋白质泛素化模式的变化。 “泛素化模式”包含多个泛素链和赖氨酸位置，这 信息比简单的泛素化事件复杂得多。此外，神经元中的泛素化模式。 AD 患者的细胞会随着疾病的进展而发生变化。没有针对症状前或病因的无创血液检测。 阿尔茨海默病的诊断除了诊断之外，还具有分子识别的能力。 AD 的特征可以帮助对患者群体进行分层，并且可以利用泛素化模式的变化 监测针对泛素蛋白酶体系统的药物的临床疗效该项目的目的是。 通过以下方式捕捉神经退行性疾病建立的初始步骤以及疾病进展的阶段 识别可以在诊断测定中常规检测到的适当的泛素化模式特征。 为此，在第一阶段，LifeSensors 开发了一种高度灵敏的方法来分离多聚泛素化 基于使用 TUBES 的 Ub 链选择性的蛋白质 - 一种亲和基质，可以选择性地结合各种 不同的多聚泛素链——与质谱 (MS) 相结合，基于以下特征识别分子特征： 在模拟神经变性和标记蛋白的神经元细胞中模拟条件。 已被鉴定；此外，还开发了方法来鉴定低水平的泛素化蛋白质并观察 正常和 AD 患者脑脊液 (CSF) 泛素化模式的变化在第二阶段。 建议扩展这些研究以检查脑脊液和血清泛素化蛋白分子特征 大约 100 名正常人和 AD 患者将与 Nicholas Seyfried 和 Allan 博士合作。 阿尔茨海默病研究中心（AD 蛋白质组学领域的领导者）的 Levey 负责翻译开发的工具 在第一阶段监测正常对照和 AD 患者脑脊液中泛素化模式的变化，并建立 生物标记物组合可区分处于不同疾病阶段的对照者和 AD 患者，从而验证标记物 利用现有的数据库和预测研究，第二阶段的成功将导致新的发现。 AD 各个阶段的泛素化特征将使稳定同位素标记版本商业化。 这些简单的测试将分层，通过质谱对这些蛋白质进行定量。 患者群体，以监测临床试验的疗效并指导治疗 AD 药物的选择和剂量。