Stanford Tissue Mapping Center

项目摘要摘要：总体 小肠和结肠是维持人体内环境稳定的关键器官。 在摄取食物时调节营养吸收。尽管这两个器官都广泛存在于 长度，不同部分在功能和细胞异质性方面存在已知差异 每一个都是。此外，肠道中任何位置的横切面都显示出成分的复杂分层。 参与吸收和分泌、肠道内容物的运动、循环和免疫。在这 提交，我们建议继续我们在斯坦福组织测绘中心(TMC)的努力 制作小肠和结肠的多模式、三维单细胞分辨率图 墙结构。这将作为社区资源，进一步研究肠道功能和 疾病。我们将在明确同意分发的情况下收集已故器官捐赠者的组织。 在HuBMAP联盟和广泛获取基因组数据共享(GDS)之间。三套 然后将使用技术。组织样本将接受单核ATAC-SEQ 和RNA-seq.这些开放的染色质和转录图谱将在空间上映射到组织 切片采用CODEX(CO-Detect by Index)多重空间免疫分析。我们会 还采用了分子地图多重荧光原位杂交(FISH) 分析，使食典数据和单核数据能够更准确地结合。由此产生的三个- 维度图将覆盖肠壁的所有层，包括上皮、肠内分泌、 血管、淋巴管、神经、免疫和肌肉细胞群对正常 大便功能。

PROJECT SUMMARY ABSTRACT: Overall The small bowel and colon are organs critical for maintaining homeostasis of the human body by mediating nutritional absorption upon the ingestion of food. Though both organs are extensive in length, there are known differences in function and cellular heterogeneity within different portions of each. Also, a cross section anywhere in the bowel reveals a complex layering of components involved in absorption and secretion, motility of gut contents, circulation, and immunity. In this submission, we propose to continue our efforts in the Stanford Tissue Mapping Center (TMC) to produce multi-modal, three-dimensional single-cell resolution maps of the small bowel and colonic wall structure. This will serve as a community resource to further study intestinal function and disease. We will collect tissues from deceased organ donors with explicit consent for distribution among the HuBMAP consortium and broad access genome data sharing (GDS). Three sets of technologies will then be employed. Tissue samples will be subjected to single-nuclei ATAC-seq and RNA-seq. These open chromatin and transcriptomic profiles will be spatially mapped to tissue sections using the CODEX (CO-Detection by indEXing) multiplexed spatial immunoassay. We will also employ the Molecular Cartography multiplexed fluorescence in situ hybridization (FISH) assay to enable more accurate integration of CODEX and single-nuclei data. The resultant three- dimensional maps will span all layers of the bowel wall and include the epithelial, enteroendocrine, vascular, lymphatic, nervous, immune, and muscular cell populations that contribute to normal bowel function.