Multi-omic 3D tissue maps for a Human BioMolecular Atlas

人类生物分子图谱的多组学 3D 组织图谱

• 批准号: 10649957
• 负责人: CLAYTON E MATHEWS
• 金额: $ 25万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 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/10649957
• 关键词: 3-Dimensional; Antibodies; Architecture; Atlases; Biological Assay; Biology; Cells; Communities; Complement; Computer software; Cytometry; Data Set; Detection; Diabetes Mellitus; Disease; Gene Expression; Goals; Health; Human; Human BioMolecular Atlas Program; Image; In Situ; Knowledge; Light; Lipids; Malignant neoplasm of pancreas; Maps; Mass Spectrum Analysis; Metabolic; Methods; Microscopy; Molecular; Nature; Oligonucleotides; Organ; Organ Procurements; Pancreas; Performance; Phase; Phosphorylation; Post-Translational Protein Processing; Process; Proteins; RNA; Resolution; Sampling; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Technology; Tissue Sample; Tissues; base; biobank; experience; human tissue; innovation; insight; interest; multidisciplinary; multiple omics; nano-string; novel; open source; protein expression; reconstruction; subcellular targeting; success; tissue mapping; tool

PROJECT ABSTRACT Spatially resolved molecular maps of mammalian organs hold significant promise in providing a deeper understanding of human organ functioning in health and disease states. Fundamental to this is an understanding how tissue organization impacts on the state of a cell and performance of its function. The overarching goal of the Human BioMolecular Atlas Program (HuBMAP) and specifically of Tissue Mapping Centers within the HuBMAP framework is to generate high-resolution three dimensional (3D) human tissue maps. Present state-of-the-art spatially-resolved tissue analysis assays (e.g. MERFISH, seq- FISH, imaging mass cytometry) utilize antibody-based or oligo probe-based approaches that require prior knowledge of the biomolecular targets to map, challenging the ability to characterize the terra incognita (i.e. the unknown) in a tissue mapping effort. Mass spectrometry (MS)-based omic mapping technologies enable unbiased detection and mapping of metabolites, lipids, and proteins (including post-translational modifications - PTMs) in situ in tissue samples with high-resolution and represents an excellent complement to highly multiplexed targeted approaches for spatially resolved tissue analysis. The overall objective of this application is to generate high-resolution, multi-omic, 3D biomolecular maps of non-diseased human organs. We will take a Google Maps-type approach with our mapping effort progressing in phases to generate reference maps at increasing resolution. First, single-cell or near-single-cell resolution MS-based mapping technologies will be used to provide an unbiased view of tissue molecular spatial architecture. Second, biomolecules of interest will be subsequently interrogated with highly multiplexed sub- cellular resolution spatial omics assays in a targeted fashion. Our focus will be on the pancreas, an essential organ important for several metabolic functions. Notably, the pancreas, despite its importance, is not one of the listed key tissues and organs currently being analyzed by the HuBMAP consortium further supporting the need to focus on this critical organ. We will employ high resolving power and high-resolution mass spectrometry-based molecular mapping platforms (LMD-nanoPOT-MS, MALDI-FTMS, nanoDESI-MS) for unbiased mapping of metabolites, lipids, and proteins (including PTMs such as phosphorylation). These MS assays will be complemented with powerful highly multiplexed targeted spatial omics assays (CODEX and NanoString GeoMx for protein and RNA respectively) and light sheet microscopy to generate high-resolution, multi-omics human tissue maps. The innovative spatially resolved multi-omic tissue maps generated will be unprecedented and the unique multi-omic datasets will provide many novel insights. The tissue mapping efforts will be supported by commercially available and open-source state-of-the-art 3D reconstruction software to create browsable 3D RNA/protein/PTM/lipid/metabolite maps of the pancreas. Undergirding the tissue characterization and 3D organ map reconstructions efforts will be a robust organ procurement, processing and distribution network. Specifically, we will: (1) Procure, process and distribute samples of normal pancreas from non-diseased donors through a robust procurement, processing, and distribution network. (2) Perform comprehensive high-resolution multi-omics tissue mapping through innovative and complementary platforms for unbiased and targeted analyses (that includes gene and protein expression, and PTM, metabolite and lipid abundances). (3) Establish browsable 3D multi-omics (RNA / protein / PTM / lipid / metabolite)-based maps of normal non-diseased pancreas; and to disseminate methods and tools to the HIVE and other TMCs.

项目摘要 哺乳动物器官的空间解析分子图在提供更深的 了解人体器官在健康和疾病状态中的功能。对此的基础是 了解组织组织如何影响细胞的状态及其功能的性能。 人类生物分子图集计划（Hubmap），特别是组织的总体目标 Hubmap框架内的映射中心是生成高分辨率三维（3D） 人体组织图。当前最新的空间分辨组织分析测定法（例如Merfish，Seq- 鱼，成像质量细胞术）利用基于抗体或基于寡探针的方法，需要先验 了解映射的生物分子靶标的知识，挑战了表征Terra Incognita的能力（即 在组织映射工作中未知）。质谱（MS）的OMIC映射技术启用 代谢物，脂质和蛋白质的无偏检测和映射（包括翻译后 修改-PTMS）原位在具有高分辨率的组织样品中，代表了出色的补体 采用高度多路复用的靶向方法进行空间分辨的组织分析。 该应用的总体目的是生成高分辨率，多摩变，3D生物分子图的 非疾病的人体器官。我们将采用Google Maps-type方法，而我们的映射工作不断发展 在阶段以增加分辨率生成参考图。首先，单细胞或近节点分辨率 基于MS的映射技术将用于提供组织分子空间的无偏见 建筑学。其次，随后将通过高度多重亚审查的生物分子进行审问 细胞分辨率的空间OMICS以目标方式进行。我们的重点将放在胰腺上，这是必不可少的 器官对几种代谢功能很重要。值得注意的是，胰腺尽管其重要性，但并不是 Hubmap Consortium目前正在分析列出的关键组织和器官，进一步支持 需要专注于这个关键的器官。我们将采用高分辨率和高分辨率质量 基于光谱法的分子映射平台（LMD-Nananopot-MS，Maldi-FTMS，纳米MS） 代谢物，脂质和蛋白质的无偏图（包括磷酸化等PTM）。这些MS 测定将与功能强大的高度多重目标空间OMICS分析（Codex和Codex和 分别用于蛋白质和RNA的纳米静电组）和光片显微镜，以产生高分辨率， 多词人类组织图。创新的空间解析的多摩尼克组织图将是 前所未有的和独特的多摩变数据集将提供许多新颖的见解。组织映射 努力将得到市售和开源的最先进的3D重建 软件可创建胰腺的可浏览3D RNA/蛋白质/PTM/脂质/代谢物图。抚摸 组织表征和3D器官图的重建工作将是强大的器官采购， 处理和分销网络。 具体而言，我们将：（1）从未养成 通过强大的采购，处理和分销网络捐助者。 （2）全面 高分辨率的多摩学组织通过无偏见的创新和互补平台映射 和靶向分析（包括基因和蛋白质表达，PTM，代谢产物和脂质丰度）。 （3）基于正常的可朗布罗3D多词（RNA / protin / ptm / PTM /脂质 /代谢物） 非胰腺胰腺；并将方法和工具传播到Hive和其他TMC。