Epithelial cell diversity in small intestine of pigs
Ontology highlight
ABSTRACT:
ORGANISM(S): Sus scrofa
PROVIDER: GSE208613 | GEO | 2023/01/18
REPOSITORIES: GEO
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Project description:Differentiation and specialisation of epithelial cells in the small intestine is regulated in two ways. First, there is differentiation along the crypt-villus axis of the intestinal stem cells into absorptive enterocytes, Paneth, goblet, tuft, enteroendocrine or M-cells, which is mainly regulated by WNT. Second, there is specialization along the cephalocaudal axis with different absorptive and digestive functions in duodenum, jejunum and ileum that is controlled by several transcription factors such as GATA4. However, so far it is unknown whether location-specific functional properties are intrinsically programmed within stem cells or if continuous signalling from mesenchymal cells is necessary to maintain the location-specific identity of the small intestine. By using the pure epithelial organoid technique, we show that region-specific gene expression profiles are conserved throughout long-term cultures of both mouse and human intestinal stem cells and correlated with differential Gata4 expression. Furthermore, the human organoid culture system demonstrates that Gata4-regulated gene expression is only allowed in absence of WNT signalling. These data show that location-specific function is intrinsically programmed in the adult stem cells of the small intestine and that their differentiation fate is independent of location-specific extracellular signals. In light of the potential future clinical application of small intestine-derived organoids, our data imply that it is important to generate GATA4-positive and GATA4-negative cultures to regenerate all essential functions of the small intestine. RNA sequencing of intestinal crypts, villi and cultured organoids derived from mouse duodenum, jejunum and ileum
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Project description:Differentiation and specialisation of epithelial cells in the small intestine is regulated in two ways. First, there is differentiation along the crypt-villus axis of the intestinal stem cells into absorptive enterocytes, Paneth, goblet, tuft, enteroendocrine or M-cells, which is mainly regulated by WNT. Second, there is specialization along the cephalocaudal axis with different absorptive and digestive functions in duodenum, jejunum and ileum that is controlled by several transcription factors such as GATA4. However, so far it is unknown whether location-specific functional properties are intrinsically programmed within stem cells or if continuous signalling from mesenchymal cells is necessary to maintain the location-specific identity of the small intestine. By using the pure epithelial organoid technique, we show that region-specific gene expression profiles are conserved throughout long-term cultures of both mouse and human intestinal stem cells and correlated with differential Gata4 expression. Furthermore, the human organoid culture system demonstrates that Gata4-regulated gene expression is only allowed in absence of WNT signalling. These data show that location-specific function is intrinsically programmed in the adult stem cells of the small intestine and that their differentiation fate is independent of location-specific extracellular signals. In light of the potential future clinical application of small intestine-derived organoids, our data imply that it is important to generate GATA4-positive and GATA4-negative cultures to regenerate all essential functions of the small intestine.
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Project description:MicroRNA-mediated regulation is critical for the proper development and function of the small intestinal epithelium. However, it is not yet known which microRNAs are expressed in which cell types of the small intestinal epithelium. To bridge this important knowledge gap, in this study we performed a comprehensive profiling analysis of microRNAs in all major epithelial cell types of the mouse small intestine. We first used flow cytometry and fluorescence activated cell sorting with multiple different reporter mouse models to isolate intestinal stem cells (ISCs), enterocytes, goblet cells, Paneth cells, enteroendocrine cells (EECs), tuft cells and secretory progenitor cells. We then subjected these cell populations to small RNA-seq. We identified highly enriched microRNA markers for every major small intestinal epithelial cell type except goblet cells. Interestingly, one of the secretory progenitor cell enriched microRNAs, miR-672, is deleted in hominin species. Several of these lineage-enriched microRNAs (LEMs) were observed to be embedded in annotated host genes. We used chromatin run-on sequencing (ChRO-seq) to determine which of these LEMs are likely co-transcribed with their host genes. We then employed single cell RNA-sequencing (scRNA-seq) in order to define the cell type specificity of the host genes, and by proxy, the LEMs as well. Finally, using four additional in vivo models, we validated that miR-375 is highly enriched in secretory progenitor cells that give rise to enteroendocrine and tuft cells and that miR-152 is a Paneth cell-specific microRNA.
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