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

项目摘要/摘要 器官特定项目组成部分 成人肺的多尺度多通道高分辨率标测 如果我们做得更好，维持正常的肺功能和诱导受损肺的修复会有明显的改善 了解器官的细胞和分子的复杂性。技术和分析方面的进步现在取得了 通过产生高信号，有可能获得以前隐藏的肺部生物学基础的详细访问 分辨率、单细胞和亚细胞、生物分子图谱合并成人类呼吸图谱 系统。HuBMAP的目标是生成交互的、可搜索的细胞类型，特别是该肺TMC的目标 以及代表转录组、蛋白质组、代谢组和SELECT的亚型注释地图和表格数据 健康肺的翻译后修饰(PTM)。最终交付成果将是将这些地图整合到 一份健康肺的参考图谱，可以与患病的肺进行比较。这样的生物分子图谱具有多个 规模、3D空间分辨率将对人类肺部生物学产生真正的变革，其后果还包括 了解全身的免疫系统和血管系统。在我们成功的HuBMAP基础上构建 第1阶段参与，我们的组织映射策略带来多种优势，包括1)成熟、强大、 组织生物信息库，2)代表整个人类肺的定向良好的组织样本，重点是 感兴趣，3)基于批量代谢组学的SnRNAseq/ATACseq双组测序和质谱学 为4)新的空间转录方法，5)空间高度多元化的化学物质的去卷积提供基础 和免疫荧光显微镜，5)一套无偏质谱、受激拉曼光谱 和先进的显微技术，所有这些都以这样一种方式结合在一起，使得空间分辨率共同定位的2D 以及跨肺组织的基因表达、代谢物、脂类、蛋白质和蛋白质修饰的3D图谱 具有单元格或接近单元格分辨率。我们将包括对40个不同的供体肺进行分析，以产生组织 用于提高检测吞吐量的微阵列。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 以细支气管束和终末-呼吸-肺泡管过渡为中心的细支气管水平 区域以及肺泡区和胸膜区。与HuBMAP蜂巢和我们的DAC合作，我们可以实现 目标是通过显微解剖、连续切片重建和厚层切片的三维成像，建立一个多尺度的 多模式高分辨率成人肺三维及图表图谱。

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.