Unknown,Transcriptomics,Genomics,Proteomics

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Adult stem cells in the small intestine are intrinsically programmed with their location-specific function

ABSTRACT: 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

ORGANISM(S): Mus musculus

SUBMITTER: Michal Mokry

PROVIDER: E-GEOD-53297 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Background and Aims: Although the zinc finger transcription factor GATA4 has been implicated in regulating jejunal gene expression, the contribution of GATA4 in controlling jejunal physiology has not been addressed. Methods: We generated mice in which the Gata4 gene was specifically deleted in the small intestinal epithelium. Measurements of plasma cholesterol and phospholipids, intestinal absorption of dietary fat and cholesterol, and gene expression were performed on these animals. Results: Mice lacking GATA4 in the intestine displayed a dramatic block in their ability to absorb cholesterol and dietary fat. Comparison of the global gene expression profiles of control jejunum, control ileum, and GATA4 null jejunum by gene array analysis demonstrated that GATA4 null jejunum lost expression of 53% of the jejunal-specific gene set and gained expression of 47% of the set of genes unique to the ileum. These alterations in gene expression included a decrease in mRNAs encoding lipid and cholesterol transporters as well as an increase in mRNAs encoding proteins involved in bile acid absorption. Conclusion: Our data demonstrate that GATA4 is essential for jejunal function including fat and cholesterol absorption and confirm that GATA4 plays a pivotal role in determining jejunal versus ileal identity. Experiment Overall Design: Total RNA was harvested from the following sources and used to Affymetrix array analysis following manufacturer defined protocols: Experiment Overall Design: control jejunum (Gata4loxP/+VilCre), 3 male mice, adult (6-8 wk) Experiment Overall Design: mutant jejunum (Gata4loxP/-VilCre), 3 male mice, adult (6-8 wk) Experiment Overall Design: control ileum (Gata4loxP/+VilCre), 3 male mice, adult (6-8 wk). Experiment Overall Design: A total of nine Mouse Genome 430_2.0 arrays were hybridized for this study. Experiment Overall Design: Jejunum was defined as 10 cm from the pyloric sphincter, and ileum was defined as 1 cm from the cecum. The animals used to harvest control jejunum and ileum were independent of each other.

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Adult stem cells in the small intestine are intrinsically programmed with their location-specific function

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