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.

抽象的 大脑是哺乳动物体内最复杂的器官。散装分析掩盖了细胞的异质性 即使在最小的大脑区域中也存在类型。多态单细胞分辨率3D-大脑的特征 组织对于创建全面的脑细胞人口普查和Altas至关重要。最近的技术 进展允许哺乳动物的单细胞转录组映射，但单细胞蛋白质组学 技术远远落后于转录组技术。缺乏高分辨率蛋白质组 BICCN财团中脑组织的表征代表 蛋白质和mRNA以更完整地了解如何将各种脑细胞组织到 独特的解剖和功能区域。该应用程序的目的是通过开发来解决这一差距 用于高分辨率3D-蛋白质组织图的可稳定质谱（MS）平台。 我们最近在技术发展方面的进步以及我们在 小鼠组织的蛋白质组映射。 AIM 1将重点放在健壮的可扩展MS平台的开发上 通过1）进一步改善样品制备，以快速处理单细胞和小组织 体素和2）利用在我们小组开发的多种破坏性技术来显着改善 检测灵敏度和样品吞吐量。预计新平台将允许可靠的量化 在单个细胞中> 3,000个蛋白质，每天约500个样品中的100个细胞中的蛋白质> 6500蛋白。 AIM 2意志 优化并演示用于小鼠拖把和人类M1的2D-蛋白质映射的可扩展MS平台 当与激光捕获组织体素的微分解时。 AIM 3将应用新平台 BICCN财团内MOP/M1的3D-蛋白质图映射，并将蛋白质组学数据与现有 转录组数据用于蛋白质组学分析。我们设想新平台将变得方便 BICCN财团中脑组织的高分辨率3D蛋白质图的必不可少的工具和 扩展BICCN工具箱。反过来，它可以做出重大贡献以提高我们对 大脑在健康和疾病中的功能。