Core B: Proteomics

核心 B：蛋白质组学

• 批准号: 10295514
• 负责人: Danielle L Swaney
• 金额: $ 16.15万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10295514
• 关键词: 17q21; Affect; Bioinformatics; Biological; Biological Assay; Cells; Chromatin Structure; Chromosomes; Communities; Complement; Data; Data Set; Detection; Development; Disease; Exhibits; Frontotemporal Dementia; Gene Expression Regulation; Gene Proteins; Genes; Genetic; Genomics; Haplotypes; Institutes; Mass Spectrum Analysis; Measures; Mediating; Molecular; Molecular Analysis; Mutation; Nerve Degeneration; Pathway interactions; Phosphorylation; Phosphorylation Site; Population; Progressive Supranuclear Palsy; Property; Protein Analysis; Proteins; Proteome; Proteomics; Protocols documentation; RNA Splicing; Reproducibility; Research; Research Project Grants; Resources; Risk; Sampling; Signal Transduction; Tauopathies; Technology; Validation; Variant; Work; base; brain cell; cell type; cost efficient; experimental study; human tissue; innovative technologies; insight; interest; member; mutant; nervous system disorder; novel; success; tau Proteins; transcriptomics

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.

项目总结（核心B：蛋白质组学） 罕见的MAPT突变长期以来一直被认为是家族性额颞叶痴呆（FTD-Tau）的原因。一 包含MAPT基因座的常见单倍型（H1）也与多发性， 散发性Tau病，包括额颞叶痴呆（FTD）和进行性核上性麻痹（PSP）， 而H2单倍型赋予降低的风险。尽管存在这种差异，但人们对 H1和H2单倍型之间风险差异背后的机制，或者这些常见的 单倍型影响罕见MAPT突变的表达。分子差异的综合分析 调解这种风险将代表神经系统疾病社区的重要和必要资源。 该核心提出的工作将在Thermo Fisher Scientific Proteomics Facility for Disease 目标发现位于加州大学旧金山分校校园的J.大卫格莱斯顿研究所。这个设施和 蛋白质组学核心将由Swaney博士领导，他是基于质谱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，使生物 发现。蛋白质组学核心的目标是提供尖端和创新的技术， 定量蛋白质组学表征调节MAPT之间差异风险的分子机制 H1和H2单倍型以可靠、可重复和具有成本效益的方式。它将支持项目1和2， 实现蛋白质组学实验，并与人体组织验证核心互动，以验证发现， 并与数据核心集成数据集作为CWOW项目成员和研究的资源 社区重要的是，它还将通过传播 新的检测和方案。这个核心的努力将集中在应用最先进的发现蛋白质组学 技术，以精确地量化蛋白质丰度和磷酸化信号的差异， 在不同的单倍型背景下，具有不同的MAPT单倍型或MAPT突变的脑细胞类型。 这些结果将提供一个蛋白质组范围内的特定蛋白质和途径的差异调节 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 一种定量少量定位于H1/H2倒位区并显示细胞类型的蛋白质的方法 以及基因表达和调控中的单倍型特异性差异。最后，我们将在蛋白质水平上进行验证 ISO-seq和转录组学分析产生的新剪接变体。