Complete Proteome of Cerebrospinal Fluid

脑脊液完整蛋白质组

• 批准号: 7016489
• 负责人: STEVEN E SCHUTZER
• 金额: $ 31.1万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-28 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7016489
• 关键词: AIDS /HIV diagnosis; AIDS dementia complex; antigen antibody reaction; biotechnology; brain disorder diagnosis; cerebrospinal fluid; clinical research; data collection methodology /evaluation; diagnosis quality /standard; genetic markers; human tissue; immune complex; liquid chromatography mass spectrometry; matrix assisted laser desorption ionization; medical complication; protein sequence; proteomics; surface enhanced laser desorption ionization; two dimensional gel electrophoresis; western blottings

DESCRIPTION (provided by applicant): The mechanism of brain dysfunction by drugs is not well characterized. Accessible objective markers to follow the influence of a substance or foreign microbe on the normal brain would be helpful. Cerebrospinal fluid (CSF) provides a liquid window to the brain. We will use a proteogenomic approach to characterize the whole proteome of normal CSF and compare it to HIV-associated dementia (HAD). Markers for this HAD alone would be useful for diagnosis and to monitor' efficacy of therapy. Although autoimmune reactivity to the brain and nervous tissue has been associated with dysfunction in several diseases, this remains an understudied area in HAD. A combined strategy using genomics, proteomics, and immunology has high potential to provide useful markers for HAD. Recently we found a significant association of antibrain antibodies, reacting to several different molecular weights of non HFV-infected brain. This proposal will use several strategies that may provide complementary and convergent results to identify markers of HAD. For Specific Aim 1, one approach is to identify the sequence of those targets of the antibrain antibodies that have been found already using the banked cerebrospinal fluid (CSF) samples from well characterized HAD patients and controls. Antigens of the molecular masses determined by the antibrain antibodies will be extracted from uninfected whole brain. These antigens will be subjected to enzymatic digestion and mass spectrometry for peptide mapping to identify the antigen by comparison to the human genome. In parallel to this, CSF antigen-antibody complexes will be probed and the antigen subjected to similar proteomic analysis. Specific Aim 2 is directed at identification of novel proteins or ratios of proteins in CSF by comparison of quantitative and qualitative proteomic profiles from HAD patients versus normals. At the very least this will provide a working database to compare disease-CSF to normals. These approaches may prove useful in investigation of this and other diseases such as substance abuse where markers are needed to monitor therapy or to illucidate interactions between drugs and microbes.

描述（由申请人提供）： 药物通过药物功能障碍的机制没有很好地表征。遵循物质或外国微生物对正常大脑的影响的无障碍物镜标记将有所帮助。脑脊液（CSF）为大脑提供了液体窗口。我们将使用一种蛋白质组方法来表征正常CSF的整个蛋白质组，并将其与HIV相关的痴呆症（HAD）进行比较。仅此而已的标记对于诊断和监测治疗功效将是有用的。尽管对大脑和神经组织的自身免疫反应性与多种疾病的功能障碍有关，但这仍然是一个研究的区域。使用基因组学，蛋白质组学和免疫学的合并策略具有很高的潜力，可以为HAT提供有用的标记。最近，我们发现了抗纤维抗体的显着关联，与非HFV感染的大脑的几种不同分子量反应。该提案将使用几种可能提供互补和收敛效果的策略来识别HAT的标记。对于特定的目标1，一种方法是识别已经发现已经使用库脑脊髓液（CSF）样品的抗振动抗体抗体靶标的序列具有患者和对照。由未感染的整个大脑中提取的由抗振动抗体抗体确定的分子质量的抗原。这些抗原将经过肽图和质谱法进行肽映射，以与人类基因组相比鉴定抗原。与此同时，将探测CSF抗原 - 抗体复合物，并进行类似的蛋白质组学分析。特定的目标2通过比较来自患者与正常的定量和定性蛋白质组学特征来鉴定CSF中新型蛋白质或蛋白质的比率。至少这将提供一个工作数据库，以将疾病-CSF与正常情况进行比较。这些方法可能被证明可用于调查这种和其他疾病，例如滥用药物，其中需要标记以监测治疗或忽略药物与微生物之间的相互作用。