A Proteomic Approach to Biomarker Discovery in Prion Disease

朊病毒病生物标志物发现的蛋白质组学方法

基本信息

批准号：
7846704
负责人：
LINGJUN LI
金额：
$ 1.93万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-05 至 2009-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7846704
关键词：
Algorithms Animal Model Animals Antibodies Bioinformatics Biological Markers Body Fluids Bovine Spongiform Encephalopathy Cerebrospinal Fluid Chronic Wasting Disease Classification Control Groups Creutzfeldt-Jakob Syndrome Detection Development Diagnosis Diagnostic Diagnostic Trial Diagnostic tests Disease Disease Outbreaks Disease Progression Early Diagnosis Family Food Formaldehyde Health Human Immunoassay Individual Infection Label Life Machine Learning Mass Spectrum Analysis Measurement Methods Mus Neuraxis Neurodegenerative Disorders Pathology Predictive Value Preparation Prion Diseases Prions Process Proteins Proteome Proteomics Rattus Research Sampling Scrapie Screening procedure Secure Sensitivity and Specificity Serum Staging Testing Therapeutic Intervention United Kingdom United States Variant Vascular blood supply base blind candidate marker cohort nervous system disorder new technology novel pre-clinical protein aminoacid sequence protein profiling tandem mass spectrometry tool

项目摘要

DESCRIPTION (provided by applicant): Transmissible spongiform encephalopathies (TSEs) are a unique family of neurodegenerative diseases of the central nervous system that are always fatal. Of the various forms of prion disease, those with the most impact upon human and animal health are Creutzfeldt-Jakob disease (CJD), bovine spongiform encephalopathy (BSE), scrapie and chronic wasting disease (CWD). The outbreaks of BSE and variant CJD (vCJD) in the United Kingdom and worldwide, including the United States, has prompted the need for rapid, reliable and inexpensive screening methods that detect TSE infection in humans and live animals. Current diagnostic tests for prion disease have, thus far, focused on detection of the causal agent of the disease, the abnormal prion protein, or individual proteins that correlate with the neurological disease. These tests are inadequate because they are post-mortem, low throughput and not sufficiently sensitive to detect infection early in the pre-clinical period. Clearly, there is an urgent need for the development of a reliable, sensitive, and specific ante-mortem diagnostic test for the pre-clinical identification of TSE-infected animals or individuals. Herein we propose the development of a new highly-sensitive mass spectrometry (MS)-based analytical platform in combination with a suite of bioinformatics tools to identify a panel of biomarkers of prion disease in body fluids collected from live animals infected with prion disease during the preclinical incubation period. The guiding hypotheses of the proposed research are: (1) The pathophysiological changes associated with prion infection will result in an altered protein profile of the cerebrospinal fluids (CSF) and/or serum proteome. (2) The combination of accurate mass measurements, isotopic labeling strategy for quantitative analyses, tandem mass spectrometry, and a suite of machine-learning algorithms will provide a unique and integrated platform to identify and characterize these diagnostic protein markers with unprecedented sensitivity and specificity. (3) Characterization of these diagnostic protein biomarkers will provide a basis for the development of an ante-mortem antibody-based screening test for prion diseases in suspected animals. To investigate these hypotheses, we propose the following specific aims: (1) To identify a panel of candidate biomarkers in the CSF and serum of animals infected with prion diseases using MS and classification algorithms. (2) To determine the identities of the candidate biomarkers by tandem MS and to obtain global quantitative changes of putative biomarkers via isotopic labeling. (3) To validate the biomarkers in blind diagnostic trials. Collectively, the proposed study will provide a basis for the development of a rapid and sensitive diagnosis of prion disease in an ante-mortem sample and it will also further our understanding of prion disease progression and pathology. This proposal explores a novel proteomic approach combining high sensitivity mass spectrometric profiling and sequencing with a set of bioinformatics tools to discover, identify, and quantify biomarkers that are indicative of prion infection. These putative biomarkers may also serve as targets for therapeutic intervention. Collectively, the proposed research will provide a basis for the development of a rapid and sensitive diagnosis of prion disease in an ante-mortem sample, enhancing our ability to secure food and blood supplies, and it will also further our understanding of prion disease progression and pathology.

描述（由申请人提供）：传染性海绵状脑病（TSE）是一种独特的中枢神经系统神经退行性疾病，通常是致命的。在各种形式的朊病毒病中，对人类和动物健康影响最大的是克雅氏病 (CJD)、牛海绵状脑病 (BSE)、痒病和慢性消耗性疾病 (CWD)。英国和包括美国在内的世界范围内 BSE 和变种克雅氏病 (vCJD) 的爆发促使人们需要快速、可靠且廉价的筛查方法来检测人类和活体动物中的 TSE 感染。迄今为止，目前对朊病毒病的诊断测试主要集中在检测疾病的致病因子、异常的朊病毒蛋白或与神经系统疾病相关的个体蛋白质。这些测试是不够的，因为它们是死后检测、通量低且不够灵敏，无法在临床前早期检测感染。显然，迫切需要开发一种可靠、灵敏且特异的生前诊断测试，用于 TSE 感染动物或个体的临床前鉴定。在此，我们建议开发一种新的基于高灵敏度质谱（MS）的分析平台，结合一套生物信息学工具，以识别从感染朊病毒病的活体动物收集的体液中的一组朊病毒病生物标志物。临床前潜伏期。本研究的指导假设是：（1）与朊病毒感染相关的病理生理变化将导致脑脊液（CSF）和/或血清蛋白质组的蛋白质谱发生改变。 (2) 精确的质量测量、用于定量分析的同位素标记策略、串联质谱和一套机器学习算法的结合将提供一个独特的集成平台，以前所未有的灵敏度和特异性来识别和表征这些诊断蛋白质标记物。 (3) 这些诊断蛋白生物标志物的表征将为开发基于生前抗体的疑似动物朊病毒病筛查试验奠定基础。为了研究这些假设，我们提出以下具体目标：（1）使用 MS 和分类算法识别感染朊病毒疾病的动物的脑脊液和血清中的一组候选生物标志物。 (2) 通过串联质谱确定候选生物标志物的身份，并通过同位素标记获得假定生物标志物的整体定量变化。 (3) 在盲法诊断试验中验证生物标志物。总的来说，这项研究将为在生前样本中快速、灵敏地诊断朊病毒病提供基础，也将进一步加深我们对朊病毒病进展和病理学的理解。该提案探索了一种新颖的蛋白质组学方法，将高灵敏度质谱分析和测序与一组生物信息学工具相结合，以发现、识别和量化指示朊病毒感染的生物标志物。这些假定的生物标志物也可以作为治疗干预的目标。总的来说，拟议的研究将为在死前样本中快速、灵敏地诊断朊病毒病奠定基础，增强我们确保食物和血液供应的能力，也将进一步加深我们对朊病毒病进展和感染的了解。病理。