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Transcriptional Profiling of Ngn3-dependent endocrine progenitors and pre-beta cells

ABSTRACT: Transcriptional Profiling of Ngn3-dependent endocrine progenitors and pre-beta cells

PROVIDER: PRJEB8315 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR739495.fastq.gz	Fastqsanger.gz
	ERR739496.fastq.gz	Fastqsanger.gz
	ERR739497.fastq.gz	Fastqsanger.gz
	ERR739498.fastq.gz	Fastqsanger.gz
	ERR739499.fastq.gz	Fastqsanger.gz

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Transcriptional Profiling of Ngn3-dependent endocrine progenitors and pre-beta cells

Project description:The goal of this study was to examine genetic networks that control endocrine cell type specification and differentiation. RNA-Seq technology was used to explore the gene expression profiles of pancreatic endocrine progenitor cells (at E10.5 and E15.5), impaired endocrine progenitor cells (at E10.5 and E15.5), and pre-beta cells (at E15.5) in mouse.

2015-01-26 | E-MTAB-3272 | biostudies-arrayexpress

NGN3 oscillatory expression controls the timing of human pancreatic endocrine differentiation

Project description:Understanding protein expression dynamics is crucial for the mechanistic understanding of cell differentiation. We investigate the dynamics and decoding of NGN3, a transcription factor critical for pancreatic endocrine development. A knock-in endogenous reporter shows that NGN3 protein oscillates with a 13-hour periodicity in human iPS-derived endocrine progenitors and is switched off as cells differentiate to β-like and pre-alpha cells. Increasing NGN3 protein stability results in one broad peak of expression instead of oscillations, with a larger peak to trough fold-change. This leads to precocious endocrine differentiation and earlier expression of key NGN3 target genes. Single-cell analysis of dynamics, mathematical modelling and experimental validation suggest that NGN3 oscillations are decoded by fold-change detection (FCD) rather than the level of expression via an incoherent feedforward loop motif (IFFL) that explains both normal and precocious differentiation. Our findings suggest that oscillatory NGN3 dynamics control the timing of differentiation, but not fate specification.

2025-03-31 | GSE281587 | GEO

Identifying new interactors of NGN3 through IP-MS

Project description:NGN3 is a transcription factor whose transient expression during pancreatic development is vital for the generation of endocrine pancreatic cells, including beta cells. NGN3 stabilisation has been shown to induce exocrine-to-endocrine cell plasticity in the murine pancreas, making it a viable target for therapies aiming to replenish beta cells after immune-mediated destruction in type 1 diabetes patients. Here, we set out to identify new interactors of NGN3 that could play a role in its post-translational regulation. We transfected HEK293A cells with HA-tagged NGN3 and carried out immunoprecipitation of the HA-tag, followed by analysis of co-immunoprecipitated interactors via LC-MS/MS.

2023-03-27 | PXD033691 | Pride

Transcription profiling of mouse Ngn3 expressing cells and endocrine pancreatic cells from wild type and Ngn3 mutant animals at day 15 of embryonic development

Project description:The basic helix-loop-helix transcription factor Neurogenin3 (Ngn3/Neurog3) is expressed in endocrine progenitor cells in the embryonic mouse pancreas. Ngn3 controls endocrine cell fate decisions. Ngn3 deficient mice do not develop any pancreatic endocrine cells, including insulin producing beta cells, and die postnatally from diabetes. Therefore, the characterization of gene expression in Ngn3-expressing cells and their progeny is of particular interest for the development of novel strategies for cell replacement therapies in type-1 diabetes. Here we describe two studies. In the first study (8 assays) we used mice where the EYFP (Enhanced Yellow fluorescent Protein) is expressed under the control of Ngn3 regulatory elements (knock add on strategy). EYFP-positive, Ngn3-expressing cells, were FACS sorted from embryonic pancreas at day 15.5 (E15.5), as well as EYFP-negative cells. In the second study (6 assays) we compared wild-type and Ngn3 mutant pancreas at E15.5. All samples were hybridized to Affymetrix GeneChip Mouse Genome 430.2.0 array.

2009-12-03 | E-CBIL-48 | biostudies-arrayexpress

Transcriptional profiling of Ngn3-dependent pancreatic endocrine differentiation at E15.5

Project description:This experiment used RNA-Seq technology to explore gene expression in mouse Ngn3^GFP/+ [het] FACS sorted pancreatic cells at E15.5 (commited endocrine progenitor cells) and in Ngn3^GFP/GFP [null] at E15.5 (defective endocrine progenitor cells). This experiment is designed to understand the gene expression alteration in the endocrine lineage at different embryonic days. The aim is to understand both Ngn3 dependent and independent gene expression profiles so as to reveal the instructive signals that specfy the collective endocrine islet cell fate or specific islet cell type.

2015-10-01 | E-MTAB-3352 | biostudies-arrayexpress

Expression data from Xenopus endoderm at stage 15 following four hours of Ngn3-GR overexpression

2012-04-01 | GSE29017 | GEO

The expression order determines the pioneer functions of Ngn3 and NeuroD1 in pancreatic endocrine differentiation

Project description:This study investigates the roles of Ngn3 and NeuroD1 in pancreatic endocrine differentiation. Using a new mouse model, we mapped NGN3 binding sites by CUT&RUN and demonstrated its pioneering role in making chromatin accessible by scATAC-seq. According to conditional knock-out and over-expression experiments and scRNA-seq, we domonstrated that NGN3's pioneering function was dose tolerance, with low doses sufficing, while NeuroD1, as a conventional TF, has no significant pioneer effects under normal phenomena. But when NGN3 was absent, NeuroD1 can alternatively acted as a pioneer factor, and sequential expression of NeuroD1 ahead of Ngn3 predominantly drives α-cell generation.

2025-02-15 | GSE270915 | GEO

Expression data from Xenopus endoderm at stage 15 following four hours of Ngn3-GR overexpression

Project description:Ngn3 is a master regulator of pancreatic endocrine development. It is necessary for the creation of all endocrine cells in mice. Little is known about the genes that act downstream of the transcription factor Ngn3 in pancreas endocrine development to specify each of the endocrine lineages. As a consequence, little is known about the genes involved in early development and the specification of the beta cell. We used microarrays to identify Ngn3 downstream genes that are involved in early and ectopic beta cell development in Xenopus laevis. We overexpressed Ngn3 in the Xenopus early endoderm and analyzed the genes that are upregulated four hours after. Xenopus laevis embryos were injected with Ngn3-GR mRNA at the eight-cell stage in the two dorsal vegetal blastomeres. Embryos at stage 12 were either treated with dexamethasone (+DEX) for four hours or no DEX (-DEX). The endoderm was dissected at stage 15. We looked for genes upregulated in the +DEX samples compared to the -DEX samples.

2012-03-31 | E-GEOD-29017 | biostudies-arrayexpress

Role of Neurogenin3 phosphorylation in the reprogramming of pancreatic organoids into endocrine cells

Project description:During pancreatic development, Neurogenin3 (Ngn3) promotes the differentiation of endocrine cells, including insulin-producing β-cells. Our previous work has shown that Ngn3 phosphorylation on multiple serine-proline (SP) sites increases protein stability and endocrine differentiation (Azzarelli et al., DevCell 2017). Here, we aim to exploit our recent data to investigate whether manipulation of Ngn3 phosphostatus might improve in vitro generation of β-cells for replacement therapies in diabetes. A potential source of β-cells comes from fate conversion of exocrine pancreatic cells into the endocrine lineage, by overexpression of 3 transcription factors, Ngn3, Pdx1 and MafA. Using this approach we show that pancreatic ductal cells grown in vitro as 3D organoids can be reprogrammed to express insulin. The efficiency can be strongly enhanced by substituting wild type Ngn3 with a the un(der)phosphorylated and more active Ngn3 form (6S-A Ngn3). Here we perform transcriptomic analysis of a low number of pancreatic organoid cells expressing wild type or phosphomutant Ngn3, alone or in various combination Pdx1 and/or MafA. The analysis revealed endocrine differentiation and in particular β-like cell gene expression in the conditions where Ngn3 was in combination with both Pdx1 and MafA.

2018-12-26 | GSE114838 | GEO

The expression order determines the pioneer functions of Ngn3 and NeuroD1 in pancreatic endocrine differentiation -2

2025-02-15 | GSE271546 | GEO