Complete Proteome of Cerebrospinal Fluid

描述（由申请人提供）： 药物引起的脑功能障碍的机制尚未得到很好的表征。追踪物质或外来微生物对正常大脑的影响的可解释的客观标志物将是有帮助的。脑脊液（CSF）为大脑提供了一个液体窗口。我们将使用蛋白基因组学方法来表征正常CSF的整个蛋白质组，并将其与HIV相关性痴呆（HAD）进行比较。这种HAD的标记物单独用于诊断和监测治疗效果是有用的。尽管对脑和神经组织的自身免疫反应与几种疾病的功能障碍有关，但这仍然是HAD中研究不足的领域。使用基因组学、蛋白质组学和免疫学的联合策略很有可能为HAD提供有用的标记物。最近，我们发现抗脑抗体的显着协会，反应几个不同分子量的非HFV感染的大脑。该提案将使用几种策略，可以提供互补和收敛的结果，以确定HAD的标记。对于特定目标1，一种方法是鉴定已经使用来自充分表征的HAD患者和对照的库存脑脊液（CSF）样品发现的抗脑抗体的那些靶标的序列。从未感染的全脑中提取分子量由抗脑抗体确定的抗原。将对这些抗原进行酶消化和质谱分析，以进行肽图谱分析，从而通过与人类基因组进行比较来鉴定抗原。与此同时，将探测CSF抗原-抗体复合物，并对抗原进行类似的蛋白质组学分析。具体目标2旨在通过比较HAD患者与正常人的定量和定性蛋白质组学图谱，鉴定CSF中的新蛋白质或蛋白质比例。至少，这将提供一个工作数据库，以比较疾病-CSF与正常。这些方法可能被证明是有用的，在调查这种疾病和其他疾病，如药物滥用，其中需要标记物来监测治疗或阐明药物和微生物之间的相互作用。

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.