A scalable mass spectrometry platform for proteome mapping of brain tissues

用于脑组织蛋白质组图谱的可扩展质谱平台

• 批准号: 10370198
• 负责人: Tujin Shi
• 金额: $ 367.37万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370198
• 关键词: 3-Dimensional; Address; Anatomy; Antibodies; Area; Atlases; Brain; Brain Mapping; Brain region; Cells; Censuses; Classification; Complex; Coupled; DNA; Data; Data Discovery; Development; Dimensions; Disease; Equilibrium; Evaluation; Generations; Goals; Health; Heterogeneity; Human; Immunoassay; Ions; Knowledge; Label; Liquid Chromatography; Mammalian Cell; Maps; Mass Spectrum Analysis; Messenger RNA; Methods; Motor Cortex; Mus; Organ; Performance; Physiology; Preparation; Proteins; Proteome; Proteomics; RNA; Resolution; Sampling; Signal Transduction; Slice; Source; Technology; Thick; Tissues; Uterus; Visualization software; base; brain cell; brain tissue; cell type; detection sensitivity; experience; genome-wide; improved; insight; instrument; ionization; laser capture microdissection; multimodal data; multimodality; multiple omics; nano-electrospray; open source; pressure; proteogenomics; single cell analysis; single cell proteins; surfactant; technology development; tool; transcriptome; transcriptomics

ABSTRACT The brain is the most complex organ in the mammalian body. Bulk analysis obscures heterogeneity of cell types present even in the smallest brain regions. Multi-omics single-cell resolution 3D-characterization of brain tissue is critically important to create comprehensive brain cell censuses and altas. Recent technological advances allow for single-cell transcriptome mapping of mammalian brains, but single-cell proteomics technologies are lagging far behind transcriptomics technologies. The lack of high-resolution proteome characterization of brain tissues in the BICCN consortium represents a significant knowledge gap between protein and mRNA for achieving a more complete understanding of how diverse brain cells are organized into distinct anatomical and functional regions. The objective of this application is to address this gap by developing a robust scalable mass spectrometry (MS) platform for high-resolution 3D-proteome mapping of brain tissues. The feasibility is strongly supported by our recent progress in technology development and our experiences in proteome mapping of mouse tissues. Aim 1 will focus on the development of a robust scalable MS platform through 1) further improving sample preparation for rapid effective processing of single cells and small tissue voxels, and 2) leveraging multiple disruptive technologies developed at our group for significantly improving detection sensitivity and sample throughput. The new platform is expected to allow for reliable quantification of >3,000 proteins in single cells and >6500 proteins in 100 cells with ~500 samples per day. Aim 2 will optimize and demonstrate the scalable MS platform for 2D-proteome mapping of mouse MOp and human M1 when combined with laser capture microdissection for tissue voxels. Aim 3 will apply the new platform for 3D-proteome mapping of MOp/M1 within the BICCN consortium and integrate proteomic data with existing transcriptomic data for proteogenomic analysis. We envision that the new platform will become a convenient indispensable tool for high-resolution 3D-proteome mapping of brain tissues in the BICCN consortium and extend the BICCN toolbox. In turn, it could make substantial contributions to improve our understanding of brain function in health and disease.

摘要 大脑是哺乳动物体内最复杂的器官。批量分析掩盖了细胞的异质性 甚至在最小的大脑区域也存在。脑的多组学单细胞分辨率3D表征 组织对于创建全面的脑细胞普查和Altas至关重要。最近的技术 研究进展允许哺乳动物大脑的单细胞转录组图谱，但单细胞蛋白质组学 技术远远落后于转录组学技术。缺乏高分辨率蛋白质组 BICCN联盟中脑组织的表征代表了 蛋白质和mRNA，以便更全面地了解不同的脑细胞是如何组织成 不同的解剖和功能区域。本申请的目的是通过开发 一个强大的可扩展质谱（MS）平台，用于脑组织的高分辨率3D蛋白质组图谱。 我们最近在技术开发方面的进展和我们在以下方面的经验有力地支持了这种可行性： 小鼠组织的蛋白质组图谱。目标1将专注于开发一个强大的可扩展MS平台 通过1）进一步改进样品制备以快速有效地处理单细胞和小组织 体素，以及2）利用我们集团开发的多种颠覆性技术， 检测灵敏度和样品通量。预计新平台将允许可靠的量化 单细胞中> 3，000种蛋白质，100个细胞中>6500种蛋白质，每天约500个样品。目标2将 优化和展示可扩展的MS平台，用于小鼠MOp和人M1的2D蛋白质组图谱 当与组织体素的激光捕获显微切割相结合时。Aim 3将应用新平台， 在BICCN联盟内进行MOp/M1的3D蛋白质组作图，并将蛋白质组数据与现有的 用于蛋白基因组学分析的转录组学数据。我们设想，新平台将成为一个方便的 BICCN联盟中脑组织高分辨率3D蛋白质组图谱的不可或缺的工具， 扩展BICCN工具箱。反过来，它可以为提高我们对 健康和疾病中的大脑功能。