Differentiation to extra-embryonic primitive endoderm from naïve human ESCs on mouse embryonic fibroblasts (MEFs) or laminin-511 (LN511) substrates
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PROVIDER: PRJNA574151 | ENA |
REPOSITORIES: ENA
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Project description:Embryonic stem cells (ESCs) can exist in at least two states that transcriptionally resemble different stages of embryonic development. Naïve ESCs resemble peri-implantation stages and primed ESCs the pre-gastrulation epiblast. In mouse, primed ESCs give rise to definitive endoderm in response to pathways downstream of Nodal and Wnt signalling. However, when these cytokines are applied to naïve ESCs, they differentiate to a cell type that approximates early primitive endoderm (PrE), the blastocyst stage progenitor layer that gives rise to the extra-embryonic endoderm. Here, we apply this context dependency to human ESCs, showing that these cytokines drive the differentiation of naïve pluripotent cells to generate extra-embryonic PrE, or hypoblast, and, as in mouse, expanded as an in vitro model for naïve extra-embryonic endoderm (nEnd) in defined conditions. Consistent with observations made in mouse, human PrE differentiation is dependent on FGF signalling in vitro, and we show, that by inhibiting FGF receptor signalling, we could simplify naïve pluripotent culture such that inhibitor requirements closer resembled those used in mouse. These nEnd cultures represent stable extra-embryonic endoderm, or human hypoblast, cell lines.
2019-11-30 | GSE138013 | GEO
Project description:IgG4-related cholangitis (IRC) is the hepatobiliary manifestation of IgG4-related disease (IgG4-RD), a systemic, B-cell driven, fibroinflammatory disorder. To date, numerous autoantigens have been described including laminin 511-E8 and carbonic anhydrase (CA) enzymes (PMID 30089633, 15647194). Laminin 511-E8 is an extracellular matrix protein that has been shown to upregulate secretory components and aid in the cholangiocyte differentiation of induced pluripotent stem cells (PMID 27103433). Carbonic anhydrase enzymes catalyse the reversible hydration of carbon dioxide, thereby producing bicarbonate and a single proton. In humans there are 14 different CA isoforms with a confined subcellular distribution. All isoforms can be targeted by the pan-CA inhibitor acetazolamide, which is a registered drug for multiple indications. We aimed to investigate the role of IgG4-RD autoantigens laminin 511-E8 and CA enzymes in human cholangiocytes. Transcriptional profiling may provide insights into the underlying molecular mechanisms of these proteins on the cholangiocellular level. Thus, by RNA sequencing the transcriptomic signature of recombinant laminin 511-E8 and acetazolamide treatment was assessed in human H69 cholangiocytes.
2024-02-15 | GSE221746 | GEO
Project description:Some mouse embryonic stem cell (mESC) lines need to be maintained on feeder cells in gelatin/Std condition. To eliminate the need for feeder cells, we decided to maintain the C57BL6/J mESCs on dishes coated with Laminin-511 (LN511) enabling maintenance of mESCs without feeder cells in Std condition (Domogatskaya et al., 2008). To compare the transcriptomes of mESCs cultured on gelatin-coated dish and those cultured on Laminin-511 coated dish, we performed RNA-Seq analysis. We found that the transcriptomes of mESCs cultured on gelatin-coated dish and those cultured on LN511 coated dish showed no considerable difference in expression patterns.
2020-06-04 | GSE132591 | GEO
Project description:Laminins are major constituents of basement membranes and are essential for tissue homeostasis. Laminin-511 is highly expressed in the intestine and its absence causes severe malformation of the intestine and embryonic lethality. To understand the mechanistic role of laminin-511 in tissue homeostasis, we used RNA profiling of embryonic intestinal tissue of lama5 knock out mice combined with cell culture experiments and siRNA mediated knock down and identified a lama5 specific gene expression signature. Our data provide a mechanistic link between laminin alpha5 gene deficiency and the physiological phenotype. We show that laminin alpha5 plays a crucial role in both epithelial and mesenchymal cell behavior by regulating Wnt and PI3K signaling. We provide evidence that a laminin-511 substratum prevents chemical-induced apoptosis in a PI3K-dependent manner and that laminin-511 represses Wnt signaling. We conclude that conflicting signals are elicited in the absence of lama5, which alter cell adhesion, migration as well as epithelial and muscle differentiation. Conversely, adhesion to laminin-511 may serve as a potent regulator of the PI3K/Akt and Wnt signaling pathways that are interconnected. Thus deregulated adhesion to laminin-511 may be instrumental in diseases such as human pathologies of the gut where laminin-511 is abnormally expressed. The microarray experiments were performed in quadruple with independently prepared RNA from mutant (Laminin alpha5-deficient mouse) and wild-type embryonic intestines derived from 4 different litters. For each RNA sample, a dye-swap labeling and hybridization was performed using a common reference RNA sample corresponding to 15.5-day embryonic intestines of C57B1/6J mice.
2012-06-29 | E-GEOD-31334 | biostudies-arrayexpress
Project description:Laminins are major constituents of basement membranes and are essential for tissue homeostasis. Laminin-511 is highly expressed in the intestine and its absence causes severe malformation of the intestine and embryonic lethality. To understand the mechanistic role of laminin-511 in tissue homeostasis, we used RNA profiling of embryonic intestinal tissue of lama5 knock out mice combined with cell culture experiments and siRNA mediated knock down and identified a lama5 specific gene expression signature. Our data provide a mechanistic link between laminin alpha5 gene deficiency and the physiological phenotype. We show that laminin alpha5 plays a crucial role in both epithelial and mesenchymal cell behavior by regulating Wnt and PI3K signaling. We provide evidence that a laminin-511 substratum prevents chemical-induced apoptosis in a PI3K-dependent manner and that laminin-511 represses Wnt signaling. We conclude that conflicting signals are elicited in the absence of lama5, which alter cell adhesion, migration as well as epithelial and muscle differentiation. Conversely, adhesion to laminin-511 may serve as a potent regulator of the PI3K/Akt and Wnt signaling pathways that are interconnected. Thus deregulated adhesion to laminin-511 may be instrumental in diseases such as human pathologies of the gut where laminin-511 is abnormally expressed.
2012-06-30 | GSE31334 | GEO
Project description:Transcriptomic signature of treatment with recombinant laminin 511-E8 and acetazolamide in human H69 cholangiocytes
| PRJNA915951 | ENA
Project description:During mammalian pre-implantation development, the cells of the blastocyst’s inner cell mass differentiate into the epiblast and primitive endoderm lineages, which give rise to the fetus and extra-embryonic tissues, respectively. Extra-embryonic endoderm differentiation can be modeled in vitro by induced expression of GATA transcription factors in mouse embryonic stem cells. Here we use this GATA-inducible system to quantitatively monitor the dynamics of global proteomic changes during the early stages of this differentiation event and also investigate the fully differentiated phenotype, as represented by embryo-derived extra-embryonic endoderm (XEN) cells. Using mass spectrometry-based quantitative proteomic profiling with multivariate data analysis tools, we reproducibly quantified 2,336 proteins across three biological replicates and have identified clusters of proteins characterized by distinct, dynamic temporal abundance profiles. We first used this approach to highlight novel marker candidates of the pluripotent state and extra-embryonic endoderm differentiation. Through functional annotation enrichment analysis, we have shown that the downregulation of chromatin-modifying enzymes, the re-organization of membrane trafficking machinery and the breakdown of cell-cell adhesion are successive steps of the extra-embryonic differentiation process. Thus, applying a range of sophisticated clustering approaches to a time-resolved proteomic dataset has allowed the elucidation of complex biological processes which characterize stem cell differentiation and could establish a general paradigm for the investigation of these processes.
2015-06-13 | PXD001901 | Pride
Project description:To experimentally investigate the function of hsa-miR-511-5p in AML monoblasts, we employed miRNA mimic mediated up-regulation of miR-511 in THP1 cells and subsequently analyzed a comprehensive gene expression profile. 4 samples of two independent miRNA mimic experiments were analyzed. THP1 monoblasts were transfected with 30 nM of Ambion Pre-miR miRNA Precursors (hsa-miR-511-5p AM10237 or negative Control #1 AM17110, Life Technologies).
2016-01-16 | E-GEOD-70490 | biostudies-arrayexpress
Project description:To experimentally investigate the function of hsa-miR-511-5p in AML monoblasts, we employed miRNA mimic mediated up-regulation of miR-511 in THP1 cells and subsequently analyzed a comprehensive gene expression profile.
2016-01-16 | GSE70490 | GEO
Project description:Haemophilus ducreyi 511 Genome sequencing project
| PRJNA65441 | ENA