Advancing Proteomic Analysis of CSF in Nervous System Diseases

描述（由申请人提供）：神经系统疾病的生物标志物可以从脑脊液（CSF）的高级蛋白质组学分析中获益，但必须首先系统评价CSF收集和处理的一些基本生物学和预分析参数以及蛋白质组学生物分析的重现性和准确性。无标记差异表达谱分析涉及通过阳离子交换色谱法分离耗尽最丰富蛋白质的CSF蛋白质组的胰蛋白酶解物，然后通过液相色谱法与质谱联用在线进行分离。 将研究从腰椎穿刺到在不同温度下冷冻的时间对不同血液污染、冻融循环、昼夜采集时间和其他变量的影响。该项目将为未来的采集工作提供最佳实践，并为储存的样本开发CSF完整性测试。最佳实践将应用于三项试点病例对照研究中的生物标志物发现-额颞叶变性（FTLD），阿尔茨海默病（AD）和精神分裂症，以发现新的生物标志物并为未来的研究提供动力计算和平台。神经科学界的资源将建立一个注释的深入目录的CSF蛋白质组和研究工具，以获取有关CSF蛋白质的信息。最后，该目录将包括每种蛋白质的胰蛋白酶肽的信息，这将使学术和制药研究人员能够构建一组肽，用于选择CSF蛋白质的多重测量，而不需要通过多反应监测（MRM）进行抗体，从而促进和促进许多神经系统疾病的生物标志物发现。 公共卫生相关性：神经系统疾病的生物标志物可以从脑脊液（CSF）的高级蛋白质组学分析中受益，但必须首先系统地评估CSF收集，处理和分析的一些基本生物学和分析参数。该项目将为未来的采集提供最佳实践，并为储存的样本开发CSF完整性测试。 在额颞叶变性（FTLD）、阿尔茨海默病和精神分裂症中发现注释的CSF蛋白质组和生物标志物将加速生物标志物的发现，以加速这些主要疾病的治疗开发。

DESCRIPTION (provided by applicant): Biomarkers for diseases of the nervous system can benefit from advanced proteomic analysis of cerebral spinal fluid (CSF) but some basic biological and pre-analytical parameters of CSF collection and handling, and reproducibility and accuracy of proteomic bioanalysis must first be systematically evaluated. Label-free differential expression profiling involves separation of tryptic digests of the CSF proteome depleted of the most abundant proteins by cation exchange chromatography followed by liquid chromatography coupled online with mass spectrometry. The effect of time from lumbar puncture to freezing at different temperatures with variable blood contamination, freeze thaw cycles, circadian time of collection and other variables will be investigated. The project will generate best practices for future collections and develop a CSF Integrity test for stored samples. Best practices will be applied to biomarker discovery in three pilot case-control studies-frontotemporal lobar degeneration (FTLD), Alzheimer's disease (AD), and schizophrenia, to find new biomarkers and inform power calculations and platforms for future studies. A resource for the neuroscience community will be established with an annotated in-depth catalog of the CSF proteome and research tools to access information about CSF proteins. Finally, the catalog will include information on tryptic peptide for each proteins that will enable academic and pharma investigators to construct a panel of peptides for multiplexed measurements of CSF proteins of choice without the need for antibody by multiple reaction monitoring (MRM), and thereby promote and facilitate biomarker discovery in many diseases of the nervous system. PUBLIC HEALTH RELEVANCE: Biomarkers for diseases of the nervous system can benefit from advanced proteomic analysis of cerebral spinal fluid (CSF) but some basic biological and analytical parameters of CSF collection, handling, and analysis must first be systematically evaluated. The project will generate best practices for future collections and develop a CSF Integrity test for stored samples. Elaboration of an annotated CSF proteome and biomarker discovery in frontotemporal lobar degeneration (FTLD), Alzheimer's disease, and schizophrenia will accelerate biomarker discovery to speed therapeutic development for these major diseases.