Core B: Proteomics

项目总结(核心B：蛋白质组学) 长期以来，人们一直知道罕见的MAPT突变是家族性额颞叶痴呆(FTD-Tau)的原因之一。一个 包含MAPT基因座的常见单倍型(H1)也与多发性、 散发的对侧病变，包括额颞部痴呆(FTD)和进行性核上性瘫痪(PSP)， 而H2单倍型则降低了风险。尽管存在这种差异，但人们对 H1和H2单倍型风险差异背后的机制，或者这些共同的 单倍型影响罕见的MAPT突变的表达。分子差异的综合分析 调解这一风险将是神经障碍社区的一个重要和必要的资源。 该核心提议的工作将在Thermo Fisher疾病科学蛋白质组学设施进行 目标发现号位于加州大学旧金山分校的J.大卫·格拉德斯通学院。这个设施和 蛋白质组学核心将由斯万尼博士领导，他是一位基于质谱学的蛋白质组学研究专家 在开发和应用新的蛋白质组学方法以使生物 发现。蛋白质组学核心的目标是为 MAPT差异风险调控分子机制的蛋白质组学定量研究 H1和H2单倍型以可靠、可重现和经济高效的方式。它将通过以下方式支持项目1和2 支持蛋白质组学实验，并与人类组织验证核心进行交互，以验证发现， 并以Data Core将数据集整合为资源，供CWow项目成员和研究人员参考 社区。重要的是，它还将成为更广泛的科学界的资源，传播 新的分析方法和方案。这一核心的努力将集中在应用最先进的发现蛋白质组学 精确量化不同品种蛋白质丰度和磷酸化信号差异的技术 不同单倍型背景下具有不同MAPT单倍型或MAPT突变的脑细胞类型。 这些结果将为差异调控的特定蛋白质和途径提供蛋白质组学的全景 MAPT单倍型。我们还将用假设驱动的靶向蛋白质组学来补充这一分析 一种定量定位于H1/H2反转区域并显示细胞类型的少量蛋白质的方法 以及单倍型特异性基因表达和调控的差异。最后，我们将在蛋白质水平上进行验证 新的剪接变异体产生于ISO-SEQ和转录切割分析。

PROJECT SUMMARY (CORE B: PROTEOMICS) Rare MAPT mutations have long been known to be a cause of familial frontotemporal dementia (FTD-Tau). A common haplotype (H1) encompassing the MAPT locus is also associated with increased risk for multiple, sporadic Tauopathies, including Frontotemporal Dementia (FTD) and Progressive Supranuclear Palsy (PSP), while the H2 haplotype confers reduced risk. Despite this difference, very little is known regarding the mechanisms behind the differences in risk between the H1 and H2 haplotypes or whether these common haplotypes affect expression of rare MAPT mutations. A comprehensive analysis of the molecular differences mediating this risk will represent a significant and essential resource for the neurological disorder community. The work proposed by this core will be performed at the Thermo Fisher Scientific Proteomics Facility for Disease Target Discovery located at the J. David Gladstone Institutes at the UCSF campus. Both this facility and the Proteomics Core, will be led by Dr. Swaney, an expert in mass-spectrometry based proteomics research with a track record in the development and application of novel proteomics approaches to enable biological discoveries. The objective of the Proteomics Core is to provide cutting-edge and innovative technologies for quantitative proteomic characterization of the molecular mechanisms regulating differential risk between MAPT H1 and H2 haplotypes in a reliable, reproducible, and cost-efficient manner. It will support Projects 1 & 2 by enabling proteomics experiments, and interact with both the Human Tissue Validation Core to validate findings, and with the Data Core to integrate datasets as a resource for the CWOW project members and the research community. Importantly, it will also serve as a resource to the wider scientific community by dissemination of new assays and protocols. Efforts of this core will be focused on applying state-of-the-art discovery proteomics technologies to precisely quantify differences in protein abundance and phosphorylation signaling for a variety of brain cell types harboring different MAPT haplotypes or MAPT mutations on different haplotypic backgrounds. These results will provide a proteome-wide view of specific proteins and pathways that are differentially regulated by MAPT haplotypes. We will also complement this analysis with a hypothesis-driven targeted proteomics approach to quantify a small number of proteins mapping to the H1/H2 inversion region and exhibiting cell type and haplotype specific differences in gene expression and regulation. Lastly, we will validate at the protein level the novel splice variants resulting from the ISO-seq and transcriptomic analyses.