Core B: Proteomics

核心 B：蛋白质组学

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要（核心B：蛋白质组学）罕见的MAPT突变长期以来一直是家族性额颞痴呆（FTD-TAU）的原因。一个涵盖MAPT基因座的常见单倍型（H1）也与多重风险增加有关零星的tauopathies，包括额颞痴呆（FTD）和进行性核上麻痹（PSP），而H2单倍型却降低了风险。尽管有这种差异，但关于 H1和H2单倍型之间的风险差异背后的机制，或者这些是否常见单倍型会影响稀有MAPT突变的表达。对分子差异的全面分析调解这种风险将代表神经系统疾病社区的重要资源。该核心提出的工作将在Thermo Fisher Scientific蛋白质组学设施中进行疾病 Target Discovery位于UCSF校园的J. David Gladstone研究所。这个设施和蛋白质组学核心将由Swaney博士领导，Swaney博士是基于质谱的蛋白质组学研究专家新型蛋白质组学方法开发和应用的记录以实现生物学发现。蛋白质组学核心的目的是为分子机制的定量蛋白质组学表征调节MAPT之间的差异风险 H1和H2单倍型的可靠，可再现和成本效益。它将支持项目1和2 启用蛋白质组学实验，并与两个人体组织验证核心相互作用，以验证发现，并且数据核心将数据集集成为CWOW项目成员和研究的资源社区。重要的是，它还将通过传播来作为更广泛科学界的资源新测定和协议。该核心的努力将集中于应用最新的发现蛋白质组学精确量化蛋白质丰度和磷酸化信号的差异的技术具有不同的MAPT单倍型或在不同单倍型背景上的MAPT突变的脑细胞类型。这些结果将为特定蛋白质和途径提供差异调节的特定蛋白质和途径的视图通过MAPT单倍型。我们还将通过假设驱动的靶向蛋白质组学对这种分析进行补充量化少量蛋白质映射到H1/H2反转区域并显示细胞类型的方法以及基因表达和调节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由ISO-SEQ和转录组分析产生的新型剪接变体。