QUANTITATIVE PROTEOMICS OF ALZHEIMER'S DISEASE HUMAN BRAIN

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 虽然晚发性阿尔茨海默病(LOAD)是最常见的神经退行性疾病之一，但其分子病因还远未完全清楚。此外，LOAD没有表现出任何明显的遗传模式，使基于遗传背景的诊断变得复杂。为了研究LOAD的分子机制和与遗传背景的潜在联系，我们使用Affymetrix 500K芯片进行了全基因组SNP基因分型，并使用Illumina ref-seq 8芯片对193个神经病理正常人脑和177个LOAD脑样本进行了转录组表达分析。到目前为止，我们已经分析和描述了SNP多态与正常人大脑基因表达谱的相关性，目前正在检查LOAD系列，以寻找可能参与疾病过程的控制RNA表达的DNA变体。然而，结合蛋白质丰度测量分析这些数据是非常重要的，因为阿尔茨海默病的主要分子特征是蛋白质聚集，这表明蛋白质生物合成和降解的失调。蛋白质聚集体：老年斑和神经原纤维缠结主要由淀粉样β蛋白组成。然而，最近通过激光捕获显微切割获得的老年斑和神经原纤维缠结的LC-MS/MS蛋白质组学研究表明，这些蛋白质聚集体更加复杂，估计蛋白质数量分别为25和60。 在目前的研究中，我们建议使用高通量的Nano-LC FTICR MS对有限的正常和负载大脑中的人类蛋白质组进行定量分析，以研究遗传多态、基因表达谱和蛋白质丰度谱之间的潜在联系，重点是蛋白质聚集。定量蛋白质组图谱将是对已经收集的正常和负载人脑的基因组和转录组图谱数据的极有价值的补充。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Although late onset Alzheimer's disease (LOAD) is one of the most common neurodegenerative disorders its molecular etiology is far from being completely understood. In addition LOAD does not show any obvious inheritance pattern, complicating the diagnostics based on genetic background. To study the molecular mechanisms of the LOAD and potential links to the genetic background we have performed whole genome SNP genotyping using the Affymetrix 500K chips and transcriptome expression analysis using the Illumina ref-seq 8 chips on a series of 193 neuropathologically normal human brains and 177 samples from LOAD brains. So far we have analyzed and described the correlation of SNP polymorphism with gene expression profiles for brains from normal individuals and currently in the process of examining the LOAD series to look for DNA variants controlling RNA expression that might be involved in disease processes. However, it is very important to analyze this data in conjunction with protein abundance measurements, as the main molecular hallmark of Alzheimer's disease is the protein aggregation, which is pointing to dis-regulation of protein biosynthesis and degradation. The protein aggregates: senile plaques and neurofibrillary tangles predominantly consist of amyloid beta protein. However the recent LC-MS/MS proteomic profiling studies of senile plaques and neurofibrillary tangles obtained by laser capture microdissection have shown that those protein aggregates are more complex with estimated number of proteins ~25 and ~60, respectively. We propose for the current study to quantitatively analyze the human proteome with high throughput nano-LC FTICR MS in a limited subset normal and LOAD brains to investigate the potential links between genetic polymorphism, gene expression profiles, and protein abundance profiles with emphasis on protein aggregation. The quantitative proteome profiling is going to be a highly valuable addition to already collected data on genome and transcriptome profiling of normal and LOAD human brains.