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）： 药物导致脑功能障碍的机制尚不明确。追踪某种物质或外来微生物对正常大脑的影响的客观标记将会有所帮助。脑脊液 (CSF) 为大脑提供了一个液体窗口。我们将使用蛋白质组学方法来表征正常脑脊液的整个蛋白质组，并将其与 HIV 相关痴呆 (HAD) 进行比较。仅针对这种 HAD 的标记就可用于诊断和监测治疗效果。尽管大脑和神经组织的自身免疫反应与多种疾病的功能障碍有关，但这在 HAD 中仍然是一个尚未得到充分研究的领域。使用基因组学、蛋白质组学和免疫学的组合策略具有为 HAD 提供有用标记的巨大潜力。最近，我们发现抗脑抗体存在显着关联，对未感染 HFV 的大脑的几种不同分子量做出反应。该提案将使用多种可能提供互补和收敛结果的策略来识别 HAD 标记。对于具体目标 1，一种方法是识别抗脑抗体靶标的序列，这些抗体已使用来自充分表征的 HAD 患者和对照的脑脊液 (CSF) 样本发现。将从未感染的全脑中提取由抗脑抗体确定的分子质量的抗原。这些抗原将经过酶消化和质谱分析以进行肽图谱分析，通过与人类基因组进行比较来识别抗原。与此同时，还将探测脑脊液抗原-抗体复合物，并对抗原进行类似的蛋白质组分析。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图谱来鉴定 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来将疾病脑脊液与正常人进行比较。这些方法可能有助于调查这种疾病和其他疾病，例如药物滥用，其中需要标记物来监测治疗或阐明药物与微生物之间的相互作用。