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受益，但脑脊液收集和处理的一些基本生物学和分析前参数，以及蛋白质组学生物分析的可重复性和准确性必须首先进行系统评估。无标记差异表达谱包括通过阳离子交换色谱法分离最丰富蛋白质的脑脊液蛋白质组的色氨酸消化，然后通过液相色谱联用在线质谱法。在不同的血液污染、冻融周期、采集时间和其他变量下，从腰椎穿刺到冷冻时间的影响将被研究。该项目将为未来的收集提供最佳实践，并为储存的样品开发CSF完整性测试。最佳实践将应用于三个试点病例对照研究——额颞叶变性（FTLD）、阿尔茨海默病（AD）和精神分裂症——的生物标志物发现，以发现新的生物标志物，并为未来的研究提供功率计算和平台。将为神经科学界建立一个有注释的脑脊液蛋白质组深度目录和研究工具，以获取有关脑脊液蛋白质的信息。最后，该目录将包括每种蛋白质的色氨酸信息，这将使学术和制药研究人员能够构建一个多肽面板，用于选择CSF蛋白质的多路测量，而不需要通过多重反应监测（MRM）的抗体，从而促进和促进许多神经系统疾病的生物标志物发现。

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.