Heading towards a dead end: the role of DND1 in germ line differentiation of human iPSCs
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE174464 | GEO | 2021/10/20
REPOSITORIES: GEO
Similar Datasets
Project description:The DND microRNA-mediated repression inhibitor 1 (DND1) is a conserved RNA binding protein (RBP) and plays an important role in survival and maintenance of primordial germ cells (PGCs) and the development of the male germline in zebrafish and mice. It was shown to be expressed in human pluripotent stem cells (PSCs), PGCs, and spermatogonia, but little is known about its specific role in pluripotency and human germline development. Here we use CRISPR/Cas mediated knockout and PGC-like cell (PGCLC) differentiation in human iPSCs to analyse if DND1 (1) plays a role in maintaining pluripotency and (2) in specification of PGCLCs. We generated several clonal lines with biallelic loss of function mutations and analysed their potential to differentiate towards PGCLCs and their gene expression on RNA and protein level via bulk RNA sequencing and mass spectrometry. The generated knockout iPSCs showed no differences in pluripotency gene expression, proliferation nor trilineage differentiation potential, but yielded reduced numbers o PGCLCs compared to their parental iPSCs. RNAseq analysis in PGCLCs showed significantly reduced expression of genes associated with cellular developmental processes and cell differentiation in knockout cells, including known markers for PGCs (NANOS3, SOX17, PRDM1, EPCAM) and naïve pluripotency (TFCP2L, DNMT3L).
2021-12-10 | PXD023802 | Pride
Project description:Heading towards a dead end: the role of DND1 in germ line differentiation of human iPSCs
| PRJNA730018 | ENA
Project description:The DND microRNA-mediated repression inhibitor 1 (DND1) is a conserved RNA binding protein (RBP) that plays important roles in survival and fate maintenance of primordial germ cells (PGCs) and in the development of the male germline in zebrafish and mice. Dead end was shown to be expressed in human pluripotent stem cells (PSCs), PGCs and spermatogonia, but little is known about its specific role concerning pluripotency and human germline development. Here we use CRISPR/Cas mediated knockout and PGC-like cell (PGCLC) differentiation in human iPSCs to determine if DND1 (1) plays a role in maintaining pluripotency and (2) in specification of PGCLCs. We generated several clonal lines carrying biallelic loss of function mutations and analysed their differentiation potential towards PGCLCs and their gene expression on RNA and protein levels via RNA sequencing and mass spectrometry. The generated knockout iPSCs showed no differences in pluripotency gene expression, proliferation, or trilineage differentiation potential, but yielded reduced numbers of PGCLCs as compared with their parental iPSCs. RNAseq analysis of mutated PGCLCs revealed that the overall gene expression remains like non-mutated PGCLCs. However, reduced expression of genes associated with PGC differentiation and maintenance (e.g., NANOS3, PRDM1) was observed. Together, we show that DND1 iPSCs maintain their pluripotency but exhibit a reduced differentiation to PGCLCs. This versatile model will allow further analysis of the specific mechanisms by which DND1 influences PGC differentiation and maintenance.
| S-EPMC8519482 | biostudies-literature
Project description:Transcript profiling of dnd1 mutant grown in control condition.
2013-01-17 | E-MEXP-3768 | biostudies-arrayexpress
Project description:Nanog, a core pluripotency factor in the inner cell mass of blastocysts, is also expressed in unipotent primordial germ cells (PGC) in mice1, where its precise role is yet unclear2-4. We investigated this in an in vitro model, where naïve pluripotent embryonic stem cells (ESCs) cultured in bFGF/ActivinA develop as epiblast-like cells (EpiLCs), and gain competence for PGC-like fate5. Consequently, bone morphogenetic protein (BMP4), or ectopic expression of key germline transcription factors Prdm1/ Prdm14/ Tfap2c, directly induce PGC-like cells (PGCLCs) in EpiLCs, but not in ESCs6-8. Here we report an unexpected discovery that Nanog alone can induce PGCLCs in EpiLCs, independently of BMP4. We propose that following the dissolution of the naïve ESC pluripotency network during establishment of EpiLCs9,10, the epigenome is reset for cell fate determination. Indeed, we found genome-wide changes in NANOG binding pattern between ESCs and EpiLCs, indicating epigenetic resetting of regulatory elements. Accordingly, we show that NANOG can bind and activate enhancers of Prdm1 and Prdm14 in EpiLCs in vitro; BLIMP1 (encoded by Prdm1) then directly induces Tfap2c. Furthermore, while SOX2 and NANOG promote the pluripotent state in ESCs, they show contrasting roles in EpiLCs since Sox2 specifically represses PGCLC induction by Nanog. This study demonstrates a broadly applicable mechanistic principle for how cells acquire competence for cell fate determination, resulting in the context-dependent roles of key transcription factors during development. To characterize Nanog-induced Primordial Germ cell-like cells (PGCLCs), we performed expression analysis of Nanog-induced Day4 PGCLCs compared to male mouse Embryonic Stem Cells (mESCs) and male Day4 PGCLCs which were induced by cytokines. mESCs were maintained in N2B27 2i(CHIR99021 3 µM, PD0325901 1 µM) LIF(1000 U/ml) medium and Day4 PGCLCs were induced in GK15 medium with Nanog induction (0.7 µg/ml) or cytokines (BMP4 500 ng/ml, BMP8A 500 ng/ml, SCF 100 ng/ml, EGF 50 ng/ml and LIF 1000U/ml) as previously reported (Hayashi K et al., Cell, 2011).
2016-01-11 | E-GEOD-61924 | biostudies-arrayexpress
Project description:At present, the etiology of most non-obstructive azoospermia (NOA) remains unclear, and the patients are diagnosed as idiopathic infertility. In vitro generation of patient-specific induced pluripotent stem cells (iPSCs) is an effective approach for exploring the mechanisms of human disease. Here, we established iPSCs from patients with idiopathic NOA, and differentiated them into human germline cells both in vitro and in vivo. Compared with normal hiPSCs, the idiopathic NOA patient-specific iPSCs show less efficiency of primodial germ cell-like cells (PGCLCs) formation in vitro and poor proliferation potential in vivo. Although the transcriptomes of PGCLCs are different from that of embryonic gonadal PGCs, the expression patterns of key genes involved in human PGC specification are generally similar during PGCLC induction process from all iPSCs lines, including the activation of BLIMP1, TFAP2C, NANOS3, SOX17, and T, and the repression of PRDM14 and SOX2. Moreover, the PGCLCs derived from idiopathic NOA patient-specific iPSCs initiate epigenetic reprogramming with repression of 5mC and presence of 5hmC. These findings provide a unique foundation for future mechanism research of male infertility.
2019-02-28 | GSE126009 | GEO
Project description:Silencing of DND1 in potato leads to resistance to late blight, powdery mildew and Botrytis cinerea. At the same time, however, it reduces plant growth and causes leaf necrosis. To get knowledge on the molecular events behind the pleiotropic effect of DND1 downregulation in potato transcriptome analysis were performed on three DND1 silenced lines in comparison with the potato cultivar ‘Désirée’ as a wild-type.
2024-10-22 | E-MTAB-13824 | biostudies-arrayexpress
Project description:The present investigation was to identify transcriptomic changes of control and Dnd1-cKO PGCs at E11.5 by RNA-seq analysis. We identified 181 upregulated and 141 downregulated genes in Dnd1-cKO PGCs.
2022-12-28 | GSE218770 | GEO
Project description:The present investigation was to identify transcriptomic changes of control and Dnd1-cKO PGCs at E12.5 and E13.5 by microarray comparison. We identified 1470 upregulated and 924 downregulated genes in Dnd1-cKO PGCs.
2022-12-28 | GSE218744 | GEO
Project description:Nanog, a core pluripotency factor in the inner cell mass of blastocysts, is also expressed in unipotent primordial germ cells (PGC) in mice1, where its precise role is yet unclear2-4. We investigated this in an in vitro model, where naïve pluripotent embryonic stem cells (ESCs) cultured in bFGF/ActivinA develop as epiblast-like cells (EpiLCs), and gain competence for PGC-like fate5. Consequently, bone morphogenetic protein (BMP4), or ectopic expression of key germline transcription factors Prdm1/ Prdm14/ Tfap2c, directly induce PGC-like cells (PGCLCs) in EpiLCs, but not in ESCs6-8. Here we report an unexpected discovery that Nanog alone can induce PGCLCs in EpiLCs, independently of BMP4. We propose that following the dissolution of the naïve ESC pluripotency network during establishment of EpiLCs9,10, the epigenome is reset for cell fate determination. Indeed, we found genome-wide changes in NANOG binding pattern between ESCs and EpiLCs, indicating epigenetic resetting of regulatory elements. Accordingly, we show that NANOG can bind and activate enhancers of Prdm1 and Prdm14 in EpiLCs in vitro; BLIMP1 (encoded by Prdm1) then directly induces Tfap2c. Furthermore, while SOX2 and NANOG promote the pluripotent state in ESCs, they show contrasting roles in EpiLCs since Sox2 specifically represses PGCLC induction by Nanog. This study demonstrates a broadly applicable mechanistic principle for how cells acquire competence for cell fate determination, resulting in the context-dependent roles of key transcription factors during development. Refer to individual Series
2016-01-11 | E-GEOD-71933 | biostudies-arrayexpress