MicroRNA介导的调控对于小肠(SI)上皮的正常发育和功能至关重要。然而,尚不清楚哪些microRNA在SI上皮细胞的每种细胞类型中表达。为了弥合这一重要的知识差距,我们在小鼠SI上皮的所有主要细胞类型中进行了全面的microRNA分析.我们使用流式细胞术和荧光激活细胞分选与多个报告小鼠模型来分离肠干细胞,肠细胞,杯状细胞,潘氏细胞,肠内分泌细胞,簇细胞,和分泌祖细胞。然后我们对这些细胞群体进行小RNA测序。得到的图谱揭示了几乎所有主要细胞类型的高度富集的microRNA标记(https://sethupathy-lab。shinyapps.io/SI_miRNA/)。观察到这些谱系富集的microRNAs(LEM)中的几种嵌入在注释的宿主基因中。我们使用染色质运行测序来确定这些LEM中的哪些可能与它们的宿主基因共同转录。然后,我们进行了单细胞RNA测序,以确定宿主基因和嵌入LEM的细胞类型特异性。我们观察到分泌祖细胞中最富集的两种microRNA是miR-1224和miR-672,我们发现后者在人源基因中缺失。最后,使用几种体内模型,我们确定miR-152是Paneth细胞特异性microRNA。在这项研究中,首先,介绍了所有主要小肠上皮细胞类型的microRNA图谱(和可搜索的网络服务器)。我们已经证明了microRNAs独特地标记了几个谱系,包括肠内分泌和簇绒。小鼠分泌祖细胞关键标记的鉴定,miR-672,我们显示它在人类中被删除。我们使用几种体内模型建立了miR-152作为Paneth细胞的特异性标志物,在microRNAs方面研究不足。
MicroRNA-mediated regulation is critical for the proper development and function of the small intestinal (SI) epithelium. However, it is not known which microRNAs are expressed in each of the cell types of the SI epithelium. To bridge this important knowledge gap, we performed comprehensive microRNA profiling in all major cell types of the mouse SI epithelium. We used flow cytometry and fluorescence-activated cell sorting with multiple reporter mouse models to isolate intestinal stem cells, enterocytes, goblet cells, Paneth cells, enteroendocrine cells,
tuft cells, and secretory progenitors. We then subjected these cell populations to small RNA-sequencing. The resulting atlas revealed highly enriched microRNA markers for almost every major cell type (https://sethupathy-lab.shinyapps.io/SI_miRNA/). Several of these lineage-enriched microRNAs (LEMs) were observed to be embedded in annotated host genes. We used chromatin-run-on sequencing to determine which of these LEMs are likely cotranscribed with their host genes. We then performed single-cell RNA-sequencing to define the cell type specificity of the host genes and embedded LEMs. We observed that the two most enriched microRNAs in secretory progenitors are miR-1224 and miR-672, the latter of which we found is deleted in hominin species. Finally, using several in vivo models, we established that miR-152 is a Paneth cell-specific microRNA.NEW & NOTEWORTHY In this study, first, microRNA atlas (and searchable web server) across all major small intestinal epithelial cell types is presented. We have demonstrated microRNAs that uniquely mark several lineages, including enteroendocrine and
tuft. Identification of a key marker of mouse secretory progenitor cells, miR-672, which we show is deleted in humans. We have used several in vivo models to establish miR-152 as a specific marker of Paneth cells, which are highly understudied in terms of microRNAs.