A Spatially Resolved Molecular Atlas of Human Endothelium

摘要-器官特异性项目（内皮素） 人体循环系统在人体每个器官的正常运作中起着至关重要的作用。 由于血管相关疾病，如动脉粥样硬化，并不均匀地发生在整个 在血管树中，不同器官的内皮一定存在差异。甚至有证据表明 相邻内皮细胞的异质性。我们提出了一个器官特异性项目（OSP），将研究正常 在整个身体和不同年龄的个体之间的内皮细胞来表征这些差异。 第一步将是从广泛的器官中获取正常组织样本。组织来源将是 脑干死亡供体，这允许在伦理范围内以最短的缺血时间收集样本。 允许基因组数据共享。从这些组织的分解样本中，我们将执行 单细胞组合索引（sci-）以生成染色质可及性（sci-ATAC-seq）和转录组 （sci-RNA-seq）来自数百个样品中数百万个单细胞的数据。这些都是建立，低成本， 可扩展的方法，这将有助于鉴定在人类中普遍存在或异质的标记物， 内皮细胞分子异质性可能与多个方面的解剖分布， 内皮，包括血管与淋巴管、动脉与静脉、毛细血管与大血管、差异 器官之间、器官分区、细胞间差异等。 对解聚细胞的深度分析，我们将采用seqFISH将不同的内皮细胞在空间上联系起来， 从单个细胞核数据产生的亚群到组织学制备。例如，这将使我们能够 识别具有特定标记物的亚群，如动脉、毛细血管、静脉或淋巴。差异 器官内特定内皮细胞亚群的转录组和染色质可及性特征， 各机关和各年龄段的人都将受到统一的综合分析框架的制约。的最终产物 这种整合将包括多参数图像，其中完整的基因表达和染色质 可访问性配置文件可以为每个单元格交互式可视化。sci-ATAC-seq的偶然收集和 非内皮细胞类型的sci-RNA-seq数据可能非常有用，可用于告知和增加 其他TMC和OSP收集的数据集，以及促进空间和分子整合 对应于具有器官特异性图的内皮的数据。我们预计，我们的空间分辨地图， 转录和染色质的可及性在整个身体的正常内皮组织中将奠定 为研究血管疾病的分子基础奠定了基础。

ABSTRACT - ORGAN SPECIFIC PROJECT (Endothelium) The human circulatory system plays a vital role in the proper functioning of every organ in the human body. Because vascular related diseases, such as atherosclerosis, do not occur homogeneously throughout the vascular tree, differences in the endothelium of various organs must exist. There is even evidence of heterogeneity of adjacent endothelial cells. We propose an Organ Specific Project (OSP) that will study normal endothelium throughout the body and across individuals of different ages to characterize these differences. The first step will be to procure normal tissue samples from a wide range of organs. The tissue source will be brainstem dead donors, which allows for collection of sample with minimal ischemic time within an ethical framework allowing for genomic data sharing. From disaggregated samples of these tissues, we will perform single cell combinatorial indexing (sci-) to generate chromatin accessibility (sci-ATAC-seq) and transcriptome (sci-RNA-seq) data from millions of single cells across hundreds of samples. These are established, low-cost, scalable methods that will facilitate the identification of markers that are ubiquitous or heterogeneous in human endothelium. Molecular heterogeneity might correlate with multiple facets of the anatomical distribution of endothelium, including vascular vs. lymphatic, arteries vs. veins, capillaries vs. larger vessels, differences between organs, organ zonation, cell-to-cell differences, etc. Based on heterogeneous markers defined from deep profiling of disaggregated cells, we will employ seqFISH to spatially relate the distinct endothelial subpopulations arising from the single nuclei data to histologic preparations. For example, this will allow us to identify subpopulations with particular markers as arterial, capillary, venous, or lymphatic. Differences in the transcriptome and chromatin accessibility profiles of specific endothelial subpopulations within organs, between organs, and across ages, will be subjected to a unified, integrative analytical framework. The end product of this integration will include multiparameter images in which the complete gene expression and chromatin accessibility profile can be interactively visualized for every cell. The incidental collection of sci-ATAC-seq and sci-RNA-seq data for non-endothelial cell types may be extremely useful to inform and increase the value of the datasets collected by other TMCs and OSPs, as well as to facilitate spatial and molecular integration of the data corresponding to endothelium with organ-specific maps. We anticipate that our spatially resolved map of transcription and chromatin accessibility in normal endothelium in tissues throughout the body will lay the foundation for investigating the molecular basis for vascular diseases.