Advancing Proteomic Analysis of CSF in Nervous System Diseases

描述(申请人提供)：神经系统疾病的生物标志物可以受益于高级脑脊液蛋白质组分析，但必须首先系统地评估脑脊液收集和处理的一些基本生物学参数和分析前参数，以及蛋白质组生物分析的重复性和准确性。无标记差异表达谱包括用阳离子交换层析分离脑脊液蛋白质组的胰酶消化，去除最丰富的蛋白质，然后用液相色谱与质谱联用。将调查从腰椎穿刺点到在不同温度、不同血液污染、冻融周期、昼夜采集时间和其他变量下冻结的时间的影响。该项目将为未来的收集产生最佳做法，并为储存的样本开发一项脑脊液完整性测试。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-生物标记物的发现，以发现新的生物标记物，并为未来研究提供能量计算和平台。将为神经科学界建立一个资源，其中包括一个带注释的深入的脑脊液蛋白质组目录和研究工具，以获取关于脑脊液蛋白质的信息。最后，该目录将包括每种蛋白质的胰蛋白酶多肽的信息，这将使学术和制药研究人员能够构建一组多肽，用于对所选的脑脊液蛋白质进行多路测量，而不需要通过多反应监测(MRM)来获得抗体，从而促进和促进在许多神经系统疾病中发现生物标记物。 公共卫生相关性：神经系统疾病的生物标记物可以受益于脑脊液(CSF)的高级蛋白质组学分析，但必须首先对脑脊液收集、处理和分析的一些基本生物学和分析参数进行系统评估。该项目将为未来的收集产生最佳做法，并为储存的样本开发一项脑脊液完整性测试。在额颞叶变性(FTLD)、阿尔茨海默病和精神分裂症中注解的脑脊液蛋白质组和生物标记物的发现将加速生物标记物的发现，以加快这些重大疾病的治疗开发。

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.