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Comparison of enteroendocrine cells and pancreatic β-cells using gene expression profiling and insulin gene methylation

ABSTRACT: Comparison of enteroendocrine cells and pancreatic β-cells using gene expression profiling and insulin gene methylation

PROVIDER: PRJNA490173 | ENA |

REPOSITORIES: ENA

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Comparison of enteroendocrine cells and pancreatic β-cells using gene expression profiling and insulin gene methylation.

Project description:Various subtypes of enteroendocrine cells (EECs) are present in the gut epithelium. EECs and pancreatic β-cells share similar pathways of differentiation during embryonic development and after birth. In this study, similarities between EECs and β-cells were evaluated in detail. To obtain specific subtypes of EECs, cell sorting by flow cytometry was conducted from STC-1 cells (a heterogenous EEC line), and each single cell was cultured and passaged. Five EEC subtypes were established according to hormone expression, measured by quantitative RT-PCR and immunostaining: L, K, I, G and S cells expressing glucagon-like peptide-1, glucose-dependent insulinotropic polypeptide, cholecystokinin, gastrin and secretin, respectively. Each EEC subtype was found to express not only the corresponding gut hormone but also other gut hormones. Global microarray gene expression profiles revealed a higher similarity between each EEC subtype and MIN6 cells (a β-cell line) than between C2C12 cells (a myoblast cell line) and MIN6 cells, and all EEC subtypes were highly similar to each other. Genes for insulin secretion-related proteins were mostly enriched in EECs. However, gene expression of transcription factors crucial in mature β-cells, such as PDX1, MAFA and NKX6.1, were remarkably low in all EEC subtypes. Each EEC subtype showed variable methylation in three cytosine-guanosine dinucleotide sites in the insulin gene (Ins2) promoter, which were fully unmethylated in MIN6 cells. In conclusion, our data confirm that five EEC subtypes are closely related to β-cells, suggesting a potential target for cell-based therapy in type 1 diabetes.

| S-EPMC6209304 | biostudies-literature

Comparison of enteroendocrine cells and pancreatic β-cells using gene expression profiling and insulin gene methylation

Project description:In this study, similarities between EECs and β-cells were evaluated in detail. To obtain specific subtypes of EECs, cell sorting by flow cytometry was conducted from STC-1 cells (a heterogenous EEC line), and each single cell was cultured and passaged. Five EEC subtypes were established according to hormone expression, measured by quantitative RT-PCR and immunostaining: L, K, I, G and S cells expressing glucagon-like peptide-1, glucose-dependent insulinotropic polypeptide, cholecystokinin, gastrin and secretin, respectively. Global microarray gene expression profiles revealed a higher similarity between each EEC subtype and MIN6 cells (a β-cell line) than between C2C12 cells (a myoblast cell line) and MIN6 cells, and all EEC subtypes were highly similar to each other. Genes for insulin secretion-related proteins were mostly enriched in EECs.

2018-09-11 | GSE119747 | GEO

Peptidomics of sorted enteroendocrine cells from human organoids

Project description:A human organoid culture system was set up to grow enteroendocrine cells with a venus labeled on the glucagon gene promoter sequence. This enabled the sorting of glucagon gene positive cells from negative cells, thereby enabling the enrichment of glucagon producing cells for study. Both Venus positive and venus negative cell populations were collected and their peptidome was assessed using nano LC-MS/MS

2020-07-01 | PXD017825 | Pride

RNA-Seq analysis of mouse small intestine enteroendocrine cells

Project description:We generated knock-in mice expressing GFP under the control of the endogenous GIP (Glucose-dependent Insulinotropic Polypeptide) promoter that enable the isolation of a purified population of small intestine K cells. Using RNA-Seq, we comprehensively characterized the transcriptomes of GIP-GFP cells as well as the entire enteroendocrine lineage derived from Neurogenin3 (Ngn3)-expressing progenitors. We interrogated the whole transcriptome of FACS-isolated small intestine GIPGFP cells using high-throughput mRNA sequencing. We also obtained the global gene expression patterns of the entire enteroendocrine cell lineage as well as the non-enteroendocrine cell population, comprising enterocytes, goblet cells and Paneth cells. To achieve this, small intestine epithelial cells from male mice resulting from the breeding of Neurogenin3 (Ngn3)-Cre mice with ROSA26-LoxP-STOP-LoxP-tomato indicator mice were isolated based on Tomato fluorescence and negative staining for CD45. Due to the small cell numbers, we constructed each of the three RNA-Seq libraries (GIPGFP, Ngn3TOMATO, and Ngn3-) using a pool of equal amounts of individual RNA samples without RNA amplification.

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Short-term depletion in Drosophila enteroendocrine cells

Project description:Short-term depletion in Drosophila enteroendocrine cells

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Effect of a high fat diet on gene expression in enteroendocrine cells of the mouse jejunum

Project description:To identify the genes differentially expressed in jejunal enteroendocrine cells (EECs) under a high fat diet we purified EEC (eYFP+) and non-EEC(eYFP-) using FACS from jejunal villi of Neurog3eYFP/+ mice (Mellitzer et al, Mol. Endo 2004) fed with a normal chow or high fat diet (HFD). In Neurog3eYFP/+ mice enteroendocrine progenitors and their descendants are labeled with eYFP. We then purified the RNA and perfomed gene exspression profiling using RNAseq.

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Rescue of insulin receptor condensates in insulin resistant cells

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Robust differentiation of human enteroendocrine cells from intestinal stem cells

Project description:Robust differentiation of human enteroendocrine cells from intestinal stem cells

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Identification of functional enteroendocrine regulators by real-time single-cell differentiation mapping

Project description:Homeostatic regulation of the intestinal enteroendocrine lineage hierarchy is a poorly understood process. We resolved transcriptional changes during enteroendocrine differentiation in real-time at single-cell level using a novel knock-in allele of Neurog3, the master regulator gene briefly expressed at the onset of enteroendocrine specification. A bi-fluorescent reporter, Neurog3Chrono, measures time from the onset of enteroendocrine differentiation and enables precise positioning of single-cell transcriptomes along an absolute time axis. This approach yielded a definitive description of the enteroendocrine hierarchy and its sub-lineages, uncovered differential kinetics between sub-lineages, and revealed time-dependent hormonal plasticity in enterochromaffin and L-cells. The time-resolved map of transcriptional changes predicted multiple novel molecular regulators. Nine of these were validated by conditional knockout in mice or CRISPR-modification in intestinal organoids. Six novel candidate regulators (Sox4, Rfx6, Tox3, Myt1, Runx1t1 and Zcchc12) yielded specific enteroendocrine phenotypes. Our time-resolved single cell transcriptional map presents a rich resource to unravel enteroendocrine differentiation.

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Implement enhanced artificial “intestinal enteroendocrine L cells” via oral administration for effective treatment of type 2 diabetes

Project description:Intestinal enteroendocrine L cells, responsive for secreting glucagon-like peptide-1 (GLP-1) to normalize blood glucose level, are negatively affected when type 2 diabetes (T2D) develops. Here, we designed enhanced artificial “intestinal enteroendocrine L cells” (EcN-GLP-1@Fe-TA-Pe@CLGN) for T2D treatment via oral administration.

2024-08-09 | PXD054739 |

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