Project description:Provides a set of enriched normal colon epithelial cells to use as a baseline for disease of the colon We present a dependable expression reference data set for non-neoplastic colonic epithelial cells. An enriched population of fresh colon epithelial cells were obtained from non-neoplastic, colectomy specimens and analyzed using Affymetrix GeneChip Exon 1.0 ST arrays. The findings from this study represent the first comprehensive expression profile for non-neoplastic colonic epithelial cells reported. For demonstration purposes, we have compared the data derived from these cells to a publically available set of tumor and normal matched colon data. This analysis allowed an assessment of global gene expression alterations and demonstrated that adjacent normal tissues, with a high degree of cellular heterogeneity, are not always representative of normal cells for comparison to tumors which arise from the colon epithelium. We also examined alternative splicing events in tumors compared to normal colon epithelial cells. Our analysis of splice variants on a genome-wide scale illustrate that this is a very labor intensive procedure, requiring vigilant examination of the data. It is projected that the contribution of this set of data derived from pure colonic epithelial cells will enhance studies in colon-related disease and offer a vital baseline for studies aimed at elucidating the mechanisms of alternative splicing.
Project description:To investigate lncRNA expression in colon cancer cells in comparison with paired normal colon epithelial cells by use of lncRNA microarray.
Project description:We performed a single-cell transcriptome analysis of proximal colon epithelial cells from 4 Gpr35 KO and 4 WT female littermates mice at a steady state.
Project description:The intestinal epithelium has a high turnover and constantly renews itself through proliferation of intestinal crypt cells, which depends on insufficiently characterized signals from the microenvironment. Here we show that colonic macrophages were located directly adjacent to epithelial crypt cells in mice, where they metabolically supported epithelial cell proliferation in an mTORC1-dependent manner. Specifically, mTORC1 activation in macrophages protected against colitis-induced intestinal damage and induced the synthesis of the polyamines spermidine and spermine. Epithelial cells ingested these polyamines and rewired their cellular metabolism for optimizing proliferation and defense. Notably, spermine directly stimulated proliferation of colon epithelial cells and colon organoids. Genetic interference with polyamine production in macrophages altered global polyamine levels in the colon and modified epithelial cell proliferation. Our results suggest that macrophages act as “commensals” that provide metabolic support to promote efficient self-renewal of the colon epithelium.
Project description:The colon is densely innervated by nociceptor neurons. Single cell RNA sequencing (scRNA-seq) of colon epithelial cells was performed to analyze the role of nociceptor neurons and Ramp1 signaling in regulating colonic epithelial cells.
Project description:We established a colon assembloid system comprising epithelium and diverse subtypes of stromal cells. To explore how closely assembloids resemble the native tissue, we performed the transcriptional profiling of stromal cells from assembloids and epithelial cells from assembloids, organoids, and colon tissue at the single-cell level.
