A Spatially Resolved Molecular Atlas of Human Endothelium

抽象-器官特异性项目(内皮) 人体循环系统对人体每个器官的正常运作起着至关重要的作用。 因为血管相关的疾病，如动脉粥样硬化，并不是在整个 维管束树，不同器官的内皮细胞必然存在差异。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 全身和不同年龄段的个体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 脑干死亡供者，允许在伦理范围内以最小的缺血时间收集样本 允许基因组数据共享的框架。从这些组织的分解样本中，我们将进行 单细胞组合索引(SCI-)产生染色质可及性(SCI-ATAC-SEQ)和转录组 (SCI-RNA-SEQ)来自数百个样本的数百万个单细胞的数据。这些都是成熟的、低成本的、 可扩展的方法，将有助于识别在人类中普遍存在或不同种类的标记 内皮细胞。分子的异质性可能与心脏解剖分布的多方面相关。 内皮，包括血管与淋巴管、动脉与静脉、毛细血管与较大血管，差异 器官之间、器官分区、细胞到细胞的差异等 深入剖析分解的细胞，我们将使用seqFISH来空间关联不同的内皮细胞 从单核数据到组织学准备的亚群。例如，这将允许我们 识别具有特定标记的亚群，如动脉、毛细血管、静脉或淋巴管。中美之间的差异 器官内特定内皮细胞亚群的转录组和染色质可及性图谱 各器官以及不同年龄的人都将受到统一、综合的分析框架的影响。的最终产品 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 可以交互地可视化每个单元的辅助功能配置文件。附带收集的SCI-ATAC-SEQ和 非内皮细胞类型的SCI-RNA-SEQ数据可能对告知和增加 其他TMC和OSP收集的数据集，以及促进空间和分子集成 与具有器官特定地图的内皮相对应的数据。我们预计我们的空间分辨率地图 转录和染色质的可及性在整个身体组织的正常内皮细胞中将奠定 研究血管疾病分子基础的基础。

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.