RNA sequencing for PDX1, NGN3 and MAFA transduced iPSCs cell
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA552353 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR9636720_1.fastq.gz | Fastqsanger.gz | |||
| SRR9636720_2.fastq.gz | Fastqsanger.gz | |||
| SRR9636721_1.fastq.gz | Fastqsanger.gz | |||
| SRR9636721_2.fastq.gz | Fastqsanger.gz | |||
| SRR9636712_1.fastq.gz | Fastqsanger.gz |
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Project description:Global gene expression profiling identifies accelerated induction of genes upon PDX1, NGN3 and MAFA transduction.
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Project description:Individual pancreatic buds of crosses between Ngn3+/- and Ngn3+/- ; Pdx1::Ngn3ER TM -IRES-GFP were dissected. Pregnant females were injected intra-peritoneal with 2mg of Tamoxifen at e10.25 (9 am) and dissected at e11.25 (9 to 11 am). Most of the mesenchymal tissue was removed to reduce the tissue complexity of the sample and enrich in epithelial pancreas progenitors. Moreover, at that developmental stage, Pdx1 promoter exhibits strong and homogenous expression levels. Rescuing Ngn3 expression only 24 hours before dissection (12h before activity, due to the time to process tamoxifen into the active compound 4OH tamoxifen) enriches the gene list in direct targets of Ngn3.
2014-12-02 | E-MTAB-3139 | biostudies-arrayexpress
Project description:Induced overexpression of Pdx1 in activin-induced endoderm population resulted in the upregulation of pancreas-related genes such as insulin 1 and 2 at day 20. To enhance the developmental progression from the pancreatic bud to the formation of the endocrine lineages, we next expressed neurogenin3 (Ngn3) together with Pdx-1. Induced overexpression of Pdx1 together with Ngn3 dramatically increased Insulin 1 mRNA by day 9 differentiation. The levels of insulin 1 mRNA present in the induced EBs represented approximately 100 % of that found in insulinoma cell line, betaTC6. We also confirmed insulin and C-peptide staining by immunohistochemistry. These cells process and secrete insulin and respond to various insulin secretagogues. These inductive effects were restricted to c-kit+ endoderm enriched EB-derived populations suggesting that Pdx1/Ngn3 functions at the level of pancreatic specification of endoderm in this model. Microarray analysis showed that Pdx1/Ngn3 regulated the expression of a broad spectrum of pancreatic endocrine cell-related genes.
2010-08-24 | GSE23752 | GEO
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
Project description:Induced overexpression of Pdx1 in activin-induced endoderm population resulted in the upregulation of pancreas-related genes such as insulin 1 and 2 at day 20. To enhance the developmental progression from the pancreatic bud to the formation of the endocrine lineages, we next expressed neurogenin3 (Ngn3) together with Pdx-1. Induced overexpression of Pdx1 together with Ngn3 dramatically increased Insulin 1 mRNA by day 9 differentiation. The levels of insulin 1 mRNA present in the induced EBs represented approximately 100 % of that found in insulinoma cell line, betaTC6. We also confirmed insulin and C-peptide staining by immunohistochemistry. These cells process and secrete insulin and respond to various insulin secretagogues. These inductive effects were restricted to c-kit+ endoderm enriched EB-derived populations suggesting that Pdx1/Ngn3 functions at the level of pancreatic specification of endoderm in this model. Microarray analysis showed that Pdx1/Ngn3 regulated the expression of a broad spectrum of pancreatic endocrine cell-related genes. On day 4 of a multiday differentiation protocol, EBs were dissociated and cultured in serum-free growth medium supplemented with differentiation facots, with or without Dox (1 ug/ml). On day 6, EBs were replated on gelatin in 12-well low-cluster dishes (Nunc) to obtain non-adherent floating EBs with or without Dox (1 ug/ml) and cultured out to day 13, at which time RNA was harvested for microarray analysis.
2010-08-24 | E-GEOD-23752 | biostudies-arrayexpress
Project description:Direct lineage conversion of adult cells is a promising approach for regenerative medicine. A major challenge of lineage conversion is to generate specific subtypes of cells, closely related cells with distinct properties. The pancreatic islets contain three major hormone-secreting endocrine subtypes: insulin+ β-cells, glucagon+ α-cells, and somatostatin+ δ-cells. We previously reported that a combination of three transcription factors, Ngn3, Mafa, and Pdx1, directly reprogram pancreatic acinar cells to β-cells. We now show that acinar cells can be converted to δ-like and α-like cells by Ngn3 and Ngn3+Mafa respectively. Thus, three major islet endocrine subtypes can be derived by acinar reprogramming. Ngn3 promotes establishment of a generic endocrine state in acinar cells at the onset of reprogramming in addition to promoting δ-specification. Mafa and Pdx1 suppress δ-specification in α- and β-cell formation. These studies identify a set of defined factors whose combinatorial actions reprogram acinar cells to distinct islet endocrine subtypes in vivo.
2013-11-20 | GSE52522 | GEO
Project description:Direct lineage conversion of adult cells is a promising approach for regenerative medicine. A major challenge of lineage conversion is to generate specific subtypes of cells, closely related cells with distinct properties. The pancreatic islets contain three major hormone-secreting endocrine subtypes: insulin+ β-cells, glucagon+ α-cells, and somatostatin+ δ-cells. We previously reported that a combination of three transcription factors, Ngn3, Mafa, and Pdx1, directly reprogram pancreatic acinar cells to β-cells. We now show that acinar cells can be converted to δ-like and α-like cells by Ngn3 and Ngn3+Mafa respectively. Thus, three major islet endocrine subtypes can be derived by acinar reprogramming. Ngn3 promotes establishment of a generic endocrine state in acinar cells at the onset of reprogramming in addition to promoting δ-specification. Mafa and Pdx1 suppress δ-specification in α- and β-cell formation. These studies identify a set of defined factors whose combinatorial actions reprogram acinar cells to distinct islet endocrine subtypes in vivo. induced beta cells samples at day 10 collected for the microarray
2013-11-20 | E-GEOD-52522 | biostudies-arrayexpress
Project description:We report the application of RNA-Seq technology for transcriptome analysis on Pdx1 + Ngn3 co-transfected bmMSCs. Bioinformatics analyses indicated that clean reads of the two libraries accounted for 97.24% and 97.97% of the total reads, which indicated high quality of the raw datasets. Total 2325 differentially expressed genes (DEGs) were identified in reprogrammed bmMSCs compared to the negative control, including 1476 genes up-regulated and 849 genes down-regulated. The RNA-Seq analysis globally validated in the reprogrammed cells the expression of a large number of genes involved in pancreas development, endocrine specification and islet differentiation. To the best of our knowledge, this is the first application of RNA-Seq analysis on reprogrammed MSCs towards pancreatic cells.
2019-05-18 | GSE81864 | GEO
Project description:Three master regulatory transcription factors Pdx1, MafA and Ngn3 have the ability to transdifferentiate pancreatic acinar cells to insulin-producing beta cells in mice. BRD7552 was identified as a small-molecule inducer that can upregulate the expression of Pdx1 in PANC-1 cells by high-throughput qPCR screening. We used microarrays to illustrate the mechnism of BRD7552 for PDX1 and insulin induction.
2013-03-10 | GSE44946 | GEO
Project description:Three master regulatory transcription factors Pdx1, MafA and Ngn3 have the ability to transdifferentiate pancreatic acinar cells to insulin-producing beta cells in mice. BRD7552 was identified as a small-molecule inducer that can upregulate the expression of Pdx1 in PANC-1 cells by high-throughput qPCR screening. We used microarrays to illustrate the mechnism of BRD7552 for PDX1 and insulin induction. PANC-1 cells were treated with BRD7552 at 5uM, the mininal concentration for the maximal induction of Pdx1. The RNA was isloated at two time points 6-hour and 72-hour.
2013-03-10 | E-GEOD-44946 | biostudies-arrayexpress