Cerebrospinal Fluid Biomarkers for Alzheimer's Disease

描述（由申请人提供）：阿尔茨海默病（AD）的早期和准确诊断迫切需要生物标志物。surmed正在使用高通量、多重蛋白质组学方法检测、鉴定和量化动物模型和人脑脊液（CSF）标本中大量假定的生物标志物。目的1提出利用微尺度液相色谱和质谱（MS）技术分离脑脊液并鉴定脑脊液中多肽和蛋白质的方法。我们将使用专有的生物信息学在质谱中识别和量化代谢物，肽，蛋白质。Aim II提出确定与已知或假设参与AD的潜在病理生理过程相关的特定候选生物标志物，特别是泛素蛋白酶体系统（UPS）。我们将在啮齿动物原代神经元细胞培养中利用蛋白酶体抑制来模拟AD，因为蛋白酶体抑制被怀疑是AD病理的早期事件。我们将使用各种亲和技术来捕获由于UPS抑制而积累的肽或蛋白质。捕获的蛋白质将通过质谱鉴定，并添加到假定的生物标志物列表中。综上所述，这些研究将证明我们广泛的表型分析方法在识别和量化生物标志物方面的可行性，并为我们研究人类脑脊液样本提供有效的方法和工具。

DESCRIPTION (provided by applicant): Biomarkers are urgently needed for the early and accurate diagnosis of Alzheimer's disease (AD). SurroMed is using a high throughput, multiplexed proteomics approach to detect, identify and quantify a large number of putative biomarkers in animal models and human cerebrospinal fluid (CSF) specimens. Aim I proposes to develop methods to fractionate CSF and identify peptides and proteins in CSF using micro-scale liquid chromatography and mass spectrometry (MS). We will use proprietary bioinformatics to identify and quantify metabolites, peptides, proteins in mass spectra. Aim II proposes to identify specific candidate biomarkers related to underlying pathophysiological processes with known or hypothesized involvement in AD, specifically the ubiquitin proteasome system (UPS). We will develop conditions to simulate AD using proteasome inhibition in rodent primary neuronal cell cultures since proteasome inhibition is suspected to occur as an early event in AD pathology. We will use a variety of affinity techniques to capture peptides or proteins that accumulate due to UPS inhibition. Captured proteins will be identified by MS, and added to a list of putative biomarkers. Taken together, these studies will demonstrate the feasibility of our broad phenotypic analysis approach to identifying and quantifying biomarkers and provide us with validated methods and tools for studies of human CSF samples. PROPOSED COMMERCIAL APPLICATION: We propose to develop a global solution for proteomic analysis using an innovative technology platform for probing the molecular signatures of Alzheimer's disease (AD). The analysis will be coupled with proprietary bioinformatics to identify and validate putative biomarkers for AD. This research program will lead to commercialization of diagnostic kits, therapeutic targets, instrumentation, and bioinformatics.