Multimodal mass spectrometry imaging of mouse and human liver

小鼠和人类肝脏的多模态质谱成像

• 批准号: 10817566
• 负责人: Brent R Stockwell
• 金额: $ 15万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-10 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10817566
• 关键词: 3-Dimensional; Administrative Supplement; Antibodies; Architecture; Atlases; Cells; Cellular Structures; Chicago; Cirrhosis; Data Set; Databases; Disease; Extracellular Matrix; Fibrosis; Future; Human; Human BioMolecular Atlas Program; Illinois; Liver; Maps; Metabolic; Modality; Molecular; Multimodal Imaging; Mus; Nature; Organ; Preparation; Primary carcinoma of the liver cells; Proteome; Proteomics; Resolution; Sampling; Signal Transduction; Spatial Distribution; Structure; Time; Tissues; Universities; Variant; Vision; Visualization; cell type; data integration; extracellular; lipidome; lipidomics; mass spectrometric imaging; metabolome; metabolomics; meter; multimodality; multiple omics; programs; technology development

Project summary We request an administrative supplement to build the first spatially-resolved 3D map of the human liver integrating both cellular components and metabolites and the extracellular matrix for the first time. This collaborative effort between the HuBMAP Transformative Technology Development team led by Dr. Tian (University of Pittsburgh, Pitt) and Dr. Stockwell (Columbia University, CU) and the Demonstration Program team led by Dr. Naba (University of Illinois Chicago, UIC) is in line with HuBMAP’s vision for comprehensive organ mapping, providing an unprecedented view into liver architecture with a depth never achieved before. An extensive effort has been made to profile the extracellular matrix (ECM) composition – or matrisomes – of various organs; however, there is still lack of study on the spatial distribution of ECM components surrounding cells and contributing to functional multicellular structures, their variation in normal vs. disease, and molecular signaling/crosstalk with cells. The challenges lie in (1) the lack of panels of validated anti-ECM antibodies, (2) the difficulty in performing multiplexed mapping of multi-level biomolecules at single-cell resolution, and (3) the difficulty to integrate datasets generated using multiple “-omic” modalities in a single sample. The team led by Dr. Tian developed a mass spectrometry imaging (H2O)n>25k-GCIB-SIMS dual-SIMS workflow, integrating untargeted metabolomics, lipidomics, and targeted proteomics (up to 40 targets) on the same tissue section at subcellular spatial resolution (1 µm). Together with the team led by Dr. Stockwell, a robust multimodal imaging workflow has been established to generate a spatially-resolved atlas of liver tissue, visualizing major tissue structures, cell types, and metabolic states of cell types. Leveraging the draft of the human liver matrisome obtained by Dr. Naba and the content of MatrisomeDB, the database of ECM proteomics datasets her team created, we propose to create a spatially-resolved map of essential human liver matrisome components, along with our current multiplexed liver map at the single-cell level. This project will present a new opportunity to delineate the spatial organization of cellular and extracellular biomolecules, the definition of functional tissue units, including information on microenvironmental niches. This will allow, in the future, to define the nature of the interactions and signals established between cells and their surrounding ECM, and to begin to understand disease-associated dysregulations (e.g., fibrosis, cirrhosis, hepatocellular carcinoma).

项目概要 我们请求行政补充，以构建第一个空间分辨的 3D 人类地图 肝脏首次整合细胞成分和代谢物以及细胞外基质 HuBMAP 转型技术开发团队之间的合作努力。 Tian博士（匹兹堡大学，皮特）和Stockwell博士（哥伦比亚大学，CU）领导的团队 由 Naba 博士（伊利诺伊大学芝加哥分校，UIC）领导的示范项目团队正在 符合 HuBMAP 全面器官绘图的愿景，提供前所未有的视图 以前所未有的深度深入了解肝脏结构。 我们付出了巨大的努力来分析细胞外基质 (ECM) 的组成，或者 基质体——各种器官的基质体；然而，仍然缺乏对其空间分布的研究。 ECM 成分围绕细胞并有助于功能性多细胞结构，它们的 正常与疾病的差异，以及与细胞的分子信号传导/串扰。 (1) 缺乏经过验证的抗 ECM 抗体组，(2) 进行多重检测存在困难 在单细胞分辨率下绘制多级生物分子图谱，以及（3）整合的难度 田博士领导的团队在单个样本中使用多种“-omic”模式生成的数据集。 开发了质谱成像 (H2O)n>25k-GCIB-SIMS 双 SIMS 工作流程，集成 同一目标的非靶向代谢组学、脂质组学和靶向蛋白质组学（最多 40 个目标） 与 Stockwell 博士领导的团队一起进行亚细胞空间分辨率 (1 µm) 的组织切片。 建立了强大的多模态成像工作流程来生成空间分辨图集 肝组织，可视化主要组织结构、细胞类型和细胞类型的代谢状态。 利用纳巴博士获得的人类肝脏基质体草图和 MatrisomeDB，她的团队创建的 ECM 蛋白质组数据集数据库，我们建议创建 人类肝脏基质成分的空间分辨图，以及我们目前的 单细胞水平的多重肝脏图谱。 该项目将为描绘细胞和细胞的空间组织提供新的机会。 细胞外生物分子，功能组织单位的定义，包括信息 这将在未来定义相互作用的性质。 以及细胞与其周围 ECM 之间建立的信号，并开始了解 与疾病相关的失调（例如纤维化、肝硬化、肝细胞癌）。