Synthesis of the Multi-Modality High Resolution 3-D Atlas of Human Lung

项目概要/摘要 器官特定项目组成部分 成人肺的多尺度多模态高分辨率测绘 如果我们更好地维持正常的肺功能并诱导受损肺的修复，可以得到显着改善。 了解器官的细胞和分子复杂性。技术和分析的进步现在使得 通过产生高通量，可以详细了解以前隐藏的肺生物学基础知识 分辨率、单细胞和亚细胞、生物分子图谱合并成人类呼吸系统图谱 系统。 HuBMAP（尤其是 Lung TMC）的目标是生成交互式、可搜索的细胞类型 以及代表转录组、蛋白质组、代谢组和选择的子类型注释图谱和表格数据 健康肺部的翻译后修饰（PTM）。最终的交付成果是将这些地图集成到 可以将健康肺部与患病肺部进行比较的参考图集。这样的生物分子图谱具有多种 规模、3D 空间分辨率将真正改变人类肺部生物学，并产生影响 了解全身的免疫和血管系统。以我们成功的 HuBMAP 为基础 在第一阶段的参与中，我们的组织图谱策略带来了多种优势，包括 1) 一个成熟的、稳健的、 组织生物储存库，2) 代表整个人肺的定向良好的组织样本，重点关注以下区域 兴趣，3）基于 snRNAseq/ATACseq 双组学测序和质谱的批量代谢组学 为以下方面的反卷积提供基础：4) 新颖的空间转录组学方法，5) 空间高度多重化学方法 和免疫荧光显微镜，5) 一套无偏质谱、受激拉曼光谱 和先进的显微镜技术，全部以这种方式结合起来，以实现空间分辨率的共定位二维 肺组织中基因表达、代谢物、脂质、蛋白质和蛋白质修饰的 3D 绘图 具有单细胞或接近单细胞的分辨率。我们将对 40 个不同的供体肺部进行分析，以生成组织 微阵列以提高检测通量。我们设想实现前所未有的生物分子水平 气管、肺外和肺内支气管与支气管动脉和淋巴系统的标测， 细支气管水平，重点关注支气管血管束和终末呼吸肺泡管过渡 区域以及肺泡和胸膜区域。与 HuBMAP HIVE 和我们的 DAC 合作，我们可实现的目标 目标是通过显微切割、连续切片重建和厚切片 3D 成像，建立多尺度 成人肺的多模态高分辨率 3D 和表格图谱。

Project Summary/Abstract Organ Specific Project Component Multi-Scale Multi-Modality High Resolution Mapping of the Adult Human Lung Maintaining normal lung function and inducing repair of damaged lung can be markedly improved if we better understand the cellular and molecular complexities of the organ. Technical and analytical advances now make it possible to gain detailed access to previously hidden fundamentals of lung biology by generating high resolution, single cell and subcellular, biomolecular maps coalesced into an atlas of the human respiratory system. It is the goal of HuBMAP and specifically of this Lung TMC to generate interactive, searchable, cell type and subtype annotated maps and tabular data representing transcriptome, proteome, metabolome, and select posttranslational modifications (PTMs) of healthy lung. Ultimate deliverable will be integration of these maps into a reference atlas of healthy lung to which diseased lung can be compared. Such a biomolecular atlas with multi- scale, 3D spatial resolution will be truly transformative for human pulmonary biology with ramifications also for understanding the immune and vascular systems throughout the body. Building upon our successful HuBMAP Phase 1 engagement, our tissue mapping strategy brings multiple strengths including 1) an established, robust, tissue biorepository, 2) well oriented tissue samples representing the entire human lung with focus on regions of interest, 3) snRNAseq/ATACseq dual-omic sequencing and mass spectroscopy based bulk metabolomics to provide basis for deconvolution of, 4) novel spatial transcriptomic methods, 5) spatial highly-multiplexed chemical and immunofluorescent microscopy, 5) a suite of unbiased mass spectrometry, stimulated Raman spectroscopy and advanced microscopy technologies, all combined in such a way to enable spatially resolved co-localized 2D and 3D mapping of gene expression, metabolites, lipids, proteins, and protein modifications across lung tissues with single-cell or near single-cell resolution. We will include assays on 40 diverse donor lungs creating tissue microarrays to enhance assay throughput. We envision achieving an unprecedented level of biomolecular mapping of the trachea, extra and intra-pulmonary bronchi with bronchial artery and lymphatic systems, several levels of bronchioles with focus on broncho-vascular bundles and terminal-respiratory-alveolar duct transition regions as well as alveolar and pleural regions. Working with the HuBMAP HIVE and our DAC, our achievable goal will be, by microdissection, serial section reconstruction and 3D imaging of thick sections, a Multi-Scale Multimodality High Resolution 3-D and Tabular Atlas of the Adult Human Lung.