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MicroRNAs control the apoptotic threshold in primed pluripotent stem cells through regulation of BIM

ABSTRACT: Mammalian primed pluripotent stem cells have been shown to be highly susceptible to cell death stimuli due to their low apoptotic threshold, but how this threshold is regulated remains largely unknown. Here we identify miRNA-mediated regulation as a key mechanism controlling apoptosis in the post-implantation epiblast. Moreover, we find that three miRNA families, miR-20, miR-92 and miR-302, control the mitochondrial apoptotic machinery by fine-tuning the levels of expression of the pro-apoptotic protein BIM. These families therefore represent an essential buffer needed to maintain cell survival in stem cells that are not only primed for differentiation but also for cell death. We used microarrays to compare the gene expression profiles of Dicer conditional Epiblast stem cells (Dicer fx/fx EpiSCs, used as control cells) and Dicer deleted epiblast stem cells (Dicer-/- EpiSCs) five days after the induction of Dicer deletion

ORGANISM(S): Mus musculus

PROVIDER: GSE61157 | GEO | 2014/09/06

SECONDARY ACCESSION(S): PRJNA260371

REPOSITORIES: GEO

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MicroRNAs control the apoptotic threshold in primed pluripotent stem cells through regulation of BIM

Project description:Mammalian primed pluripotent stem cells have been shown to be highly susceptible to cell death stimuli due to their low apoptotic threshold, but how this threshold is regulated remains largely unknown. Here we identify miRNA-mediated regulation as a key mechanism controlling apoptosis in the post-implantation epiblast. Moreover, we find that three miRNA families, miR-20, miR-92 and miR-302, control the mitochondrial apoptotic machinery by fine-tuning the levels of expression of the pro-apoptotic protein BIM. These families therefore represent an essential buffer needed to maintain cell survival in stem cells that are not only primed for differentiation but also for cell death. We used microarrays to compare the gene expression profiles of Dicer conditional Epiblast stem cells (Dicer fx/fx EpiSCs, used as control cells) and Dicer deleted epiblast stem cells (Dicer-/- EpiSCs) five days after the induction of Dicer deletion Dicer fx/fx EpiSCs were left untreated (control cells) or treated with 0.3uM of 4-OH-Tamoxifen for three days and without Tamoxifen for two further days, until day 5 when RNA was extracted and used for microarray analysis. Three independent deletion experiments including a Dicer fx/fx sample and a Dicer -/- sample were analyzed as biological replicates.

2014-09-06 | E-GEOD-61157 | biostudies-arrayexpress

Gene expression profile of Epiblast stem cells

Project description:To characterize the EpiSCs (Epiblast stem cells) we established in our lab, we have employed whole genome microarray expression profiling as a platform to distinguish the EpiSCs (primed state) from ES cells (naïve state).

2018-11-30 | GSE75660 | GEO

Formative pluripotent stem cells show features of epiblast cells poised for gastrulation

Project description:The pluripotency of mammalian early and late epiblast could be recapitulated by naïve embryonic stem cells (ESCs) and primed epiblast stem cells (EpiSCs), respectively. However, these two states of pluripotency may not be sufficient to reflect the full complexity and developmental potency of the epiblast during mammalian early development. Here we report the establishment of self-renewing formative pluripotent stem cells (fPSCs) which manifest features of epiblast cells poised for gastrulation. fPSCs can be established from different mouse ESCs, pre-/early-gastrula epiblasts and induced PSCs. Similar to pre-/early-gastrula epiblasts, fPSCs show the transcriptomic features of formative pluripotency, which are distinct from naïve ESCs and primed EpiSCs. fPSCs show the unique epigenetic states of E6.5 epiblast, including the super-bivalency of a large set of developmental genes. Just like epiblast cells immediately before gastrulation, fPSCs can efficiently differentiate into three germ layers and primordial germ cells (PGCs) in vitro. Thus, fPSCs highlight the feasibility of using PSCs to explore the development of mammalian epiblast.

2021-02-19 | GSE154290 | GEO

An alternative pluripotent state confers interspecies chimaeric competency [WGBS]

Project description:In this study, we show that by simple modulation of extrinsic signaling pathways, a new class of pluripotent stem cells, referred to as region selective epiblast stem cells (rsEpiSCs), could be efficiently derived from different stages of the early embryo. rsEpiSCs share features of primed pluripotency yet are distinct from EpiSCs in their molecular characteristics and ability to colonize post-implantation embryos. We performed whole-genome bisulfite sequencing (WGBS) experiments to compare the DNA methylation landscapes of conventional EpiSCs and rsEpiSCs. Compare the DNA methylation profiles in 2 pluripotent stem cell types (LP-EpiSCs and conventional EpiSCs) in mouse. Two replicates are examined for each cell type.

2015-05-06 | E-GEOD-63568 | biostudies-arrayexpress

RNA-seq of wild-type and PGR (progesterone receptor) knockout pluripotent mouse EpiSCs (epiblast stem cells) and differentiated samples derived from these cell lines (primitive streak and mesoderm)

Project description:EpiSCs (epiblast stem cells) are murine pluripotent stem cells derived from the epiblast of a post-implantation embryo. These cells are primed to differentiate towards embryonic germ layers and are a useful model to study these early developmental events. PGR (progesterone receptor) is a nuclear receptor described mainly in the context of fertility and breast cancer. We noticed that it is also a TF that is expressed in primed EpiSCs, but not in naive mESCs and is further upregulated during mesoderm specification. We hypothesized that it is involved in differentiation to primitive streak and mesoderm progenitors. To test this hypothesis we generated EpiSCs with a genetic knockout of PGR using CRISPR-Cas9-mediated knock-in of an antibiotic resistance cassette. Differentiation of the wild-type and knockout cells and comparison of the different cell types allowed us to conclude that PGR is important for acquiring extraembryonic mesoderm fate and modulates cardiac differentiation, specifically it ensures that correct pools of FHF (first heart field) and SHF (second heart field) are being produced. Our study sheds light on previously unappreciated role of PGR in early embryonic development.

2022-11-30 | E-MTAB-12104 | biostudies-arrayexpress

An alternative pluripotent state confers interspecies chimaeric competency [Microarray]

Project description:Here we show that by simple modulation of extrinsic signaling pathways, a new class of pluripotent stem cells, referred to as intrinsic state-epiblast stem cells (IS-EPIs), could be efficiently derived from different stages of the early embryo. IS-EPIs share features of primed pluripotency yet are distinct from EpiSCs in their molecular characteristics and ability to colonize post-implantation embryos. We performed Microarray analysis and compared global gene expression pattern among amplified RNA samples from ESCs, EpiSCS, IS-EPIs as well as in vivo isolated Epiblasts. Examination of global gene expression profiles multiple pluripotent stem cell types

2015-05-06 | E-GEOD-60523 | biostudies-arrayexpress

Zfp281 Coordinates Opposite Functions of Tet1 and Tet2 for Alternative Pluripotent States [ChIP-Seq]

Project description:Pluripotent cell identity comprises a spectrum of cell states including naive and primed states, which are typified by mouse embryonic stem cells (ESCs) and epiblast-derived stem cells (EpiSCs), respectively. Here we define a pluripotent cell fate (PCF) gene signature based on RNA-seq analysis associated with naive and primed pluripotency acquisition, and identify Zfp281 as a key transcriptional regulator for primed pluripotency and also as a barrier to achieve the naive pluripotency of both mouse and human ESCs.

2016-06-28 | GSE81042 | GEO

Zfp281 Coordinates Opposite Functions of Tet1 and Tet2 for Alternative Pluripotent States [RNA-Seq]

2016-06-28 | GSE81044 | GEO

An alternative pluripotent state confers interspecies chimaeric competency [WGBS]

2015-05-06 | GSE63568 | GEO

Capture of mouse and human stem cells with features of formative pluripotency [mouse, RNA-Seq 2]

Project description:Pluripotent cells emerge as a naïve founder population in the blastocyst, acquire capacity for germline and soma formation, and then undergo lineage priming. Mouse embryonic stem (ES) cells and epiblast stem cells (EpiSCs) respectively represent the initial naïve and final primed phases of pluripotency. Here we investigated the intermediate formative stage. Using minimal exposure to specification cues, we derived stem cells from formative mouse epiblast. Unlike ES cells or EpiSCs, formative stem (FS) cells responded directly to germ cell induction. They colonised somatic tissues and germline in chimaeras. Whole transcriptome analyses showed similarity to pre-gastrulation formative epiblast. Signal responsiveness and chromatin accessibility features reflect lineage capacitation. Furthermore, FS cells showed distinct transcription factor dependencies, relying critically on Otx2. Finally, FS cell culture conditions applied to human naïve cells or embryos supported expansion of similar stem cells, consistent with a conserved staging post on the trajectory of mammalian pluripotency.

2020-12-04 | GSE131555 | GEO

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