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）：可传播的海绵状脑病（TSE）是中枢神经系统的独特神经退行性疾病家族，总是致命的。在各种形式的prion病中，对人类和动物健康的影响最大的人是克鲁特兹菲尔特 - 贾科布疾病（CJD），牛海绵状脑病（BSE），scrapie和慢性浪费疾病（CWD）。包括美国在内的英国和全世界的BSE和变体CJD（VCJD）的爆发促使人们需要快速，可靠和廉价的筛查方法，这些方法检测到人类和活体动物中TSE感染。到目前为止，目前针对prion病的诊断测试重点是检测该疾病的因果因子，异常的prion蛋白或与神经系统疾病相关的个体蛋白质。这些测试是不足的，因为它们是验尸，低通量，并且不足以在临床前的早期检测感染。显然，迫切需要开发可靠，敏感和特定的事前诊断测试，以便对TSE感染的动物或个体进行临床前鉴定。本文中，我们建议开发一种新的高度敏感质谱法（MS）的分析平台，并结合一系列生物信息学工具，以鉴定临床前孵化期间从感染的病毒疾病感染的活体动物中收集的人体流体中的prion病疾病的生物标志物。提出的研究的指导假设是：（1）与prion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改变。（2）准确的质量测量，定量分析的同位素标记策略，串联质谱法和一系列机器学习算法的组合将提供一个独特而集成的平台，以识别和表征这些诊断蛋白质标记物具有前所未有的敏感性和特定性。（3）这些诊断蛋白生物标志物的表征将为可疑动物中的基于验尸抗体的筛查测试提供基础。为了研究这些假设，我们提出了以下特定目的：（1）确定CSF中的一组候选生物标志物和使用MS和分类算法感染的动物的动物的血清。（2）通过串联MS确定候选生物标志物的身份，并通过同位素标记获得推定生物标志物的全球定量变化。（3）在盲诊断试验中验证生物标志物。拟议的研究总的来说，将为在坦克（Ante Mortem）样本中开发快速，敏感的病毒疾病诊断的基础，这也将进一步了解我们对prion疾病进展和病理学的理解。该建议探讨了一种新型的蛋白质组学方法，该方法将高灵敏度质谱分析和测序与一组生物信息学工具结合在一起，以发现，识别和量化表明prion感染的生物标志物。这些推定的生物标志物也可以作为治疗干预的靶标。总的来说，拟议的研究将为在坦克验尸样本中开发快速和敏感的诊断疾病的诊断提供基础，增强了我们确保食品和血液供应的能力，这也将进一步了解我们对prion疾病的进展和病理学的理解。