Hypoxia directs human villous CTB differentiation towards the EVT lineage in a HIF-dependent manner.
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE80995 | GEO | 2017/02/01
SECONDARY ACCESSION(S): PRJNA320254
REPOSITORIES: GEO
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Project description:Seven patient paired primary human HLA-G+ extravillous trophoblasts (EVT) and Villous trophoblasts (VT) obtained from 1st trimester (7-9 weeks) villous tissue were obtained. RNA was isolated directly after isolation and purification using FACS sort for CD45-HLA-G+ (EVT) and CD45-HLA-G-EGFR1+ (VT) fractions. Expression profiles were compared to two samples of the choriocarcinoma cell line JEG-3 and four samples of decidual stromal cells (DSC) at passage 2 after cell culture.
2015-05-15 | E-MTAB-3217 | biostudies-arrayexpress
Project description:Invasion of cytotrophoblasts into uterine tissues is essential for placental development. To identify molecules regulating trophoblast invasion, mRNA signatures of purified villous (CTB, poor invasiveness) and extravillous (EVT, high invasiveness) trophoblasts isolated from first trimester human placentae and villous explant cultures, respectively, were compared using GeneChip analyses yielding 991 invasion/migration related transcripts. Several genes involved in physiological and pathologic cell invasion, including ADAM-12,-19,-28 as well as Spondin-2, were upregulated in EVT. Pathway prediction analyses identified several functional modules associated with either the invasive or the non-invasive trophoblast phenotype. One of the genes which were downregulated in the invasive mRNA pool, heme oxygenase-1 (HO-1), was selected for functional analyses. Real-time PCR analyses, Western blottting, and immunofluorescene of first trimester placentae and differentiating villous explant cultures demonstrated downregulation of HO-1 in invasive EVT as compared to CTB. Modulation of HO-1 expression in loss-of as well as gain-of function cell models (BeWo and HTR8/SVneo, respectively) demonstrated an inverse relationship of HO-1 expression with trophoblast migration in transwell and wound healing assays. Importantly, HO-1 expression led to an increase in protein levels and activity of the nuclear hormone receptor PPARgamma. Pharmacological inhibition of PPARgamma abrogated the inhibitory effects of HO-1 on trophoblast migration. Collectively, our results demonstrate that gene expression profiling of EVT and CTB can be used to unravel novel regulators of cell invasion. Accordingly, we identify heme oxygenase-1 as a negative regulator of trophoblast motility acting via upregulation of PPARgamma. Experiment Overall Design: To identify genes potentially regulating cell invasion trophoblast cells of early human gestation with distinct invasive properties were profiled. Experiment Overall Design: Distinct gene expression signatures of highly invasive EVT (n = 6) and poorly invasive CTB (n = 5) of different first trimester placentae using Affymetrix U133A GeneChips interrogating >20,000 genes were determined.
2009-01-30 | E-GEOD-9773 | biostudies-arrayexpress
Project description:Trophoblast is the primary epithelial cell type in the placenta, a transient organ required for proper fetal growth and development. We have identified a CDX2+/p63+ cytotrophoblast (CTB) subpopulation in the early post-implantation human placenta, which is significantly reduced later in gestation. CTB differentiate into different trophoblast subtypes, which are responsible for gas/nutrient exchange (syncytiotrophoblast/STB) and invasion and maternal vascular remodeling (extravillous trophoblast/EVT). Study of early human placental development is severely hampered by lack of a representative trophoblast stem cell (TSC) model, with the capacity for self-renewal and the ability to differentiate into both STB and EVT. We describe a reproducible protocol, using defined media containing BMP4, by which human embryonic stem cells (hESC) can be differentiated into CDX2+/p63+ CTB-like cells. These cells can be replated to further differentiate into STB- and EVT-like cells, based on marker expression, hormone secretion and invasive ability. Differentiation of hPSC-derived CTB in hypoxia leads to reduced hCG secretion and STB-associated gene expression, instead inducing EVT differentiation in a hypoxia-inducible factor-dependent manner. Human embryonic stem cells (hESC) (WA09/H9) were maintained on Geltrex-coated plates (BD Biosciences ) in StemPro (Thermo Fisher) + bFGF (12 ng/ml). Undifferentiated hESC (D-2) were switched to minimal media (EMIM (Erb et al., 2011) containing KO DMEM/F12 (Gibco), 1% Insulin-Transferrin-Selenium Mix (Sigma-Aldrich), 1% NEAA (Gibco), 2mM L-Glutamine (Corning), 0.1mM 2-mercaptoethanol (Gibo), and 2% BSA (Gemini Bio Products)) for 2 days then treated with BMP4 (StemRD, 10ng/ml) for 3 days in minimal media+BMP4 (D3). At D3, cells were replated onto Geltrex in Feeder Conditioned Media +BMP4 and cultured in normoxia (20% O2) or hypoxia (2% O2) for 2 days (D3+2) to assess the effect of hypoxia. Cells were infected with lentiviral shRNA for shScramble (control) or shARNT, the beta subunit of the Hypoxia Inducible Factor (HIF) complex to assess the effect of ARNT knockdown. Each sample includes biological triplicates.
2016-06-16 | E-GEOD-76090 | biostudies-arrayexpress
Project description:Comparison of genes associated with the EMT between cytotrophoblast cells (CTB) and extravillous trophoblast cells (EVT) from normal third trimester placenta and abnormally invasive placenta (AIP)
2018-01-01 | GSE104350 | GEO
Project description:Comparison of genes associated with the EMT between undifferentiated cytotrophoblast cells (CTB) and differentiated extravillous trophoblast cells (EVT) from third trimester human placenta. Cells isolated from control (placenta previa) and cases (preeclampsia). Cells isolated by immunomagnetic separation using anti-integrin beta4 antibody to purify CTB and anti-HLA-G antibody to purify EVT.
2022-02-26 | GSE188313 | GEO
Project description:Trophoblast is the primary epithelial cell type in the placenta, a transient organ required for proper fetal growth and development. We have identified a CDX2+/p63+ cytotrophoblast (CTB) subpopulation in the early post-implantation human placenta, which is significantly reduced later in gestation. CTB differentiate into different trophoblast subtypes, which are responsible for gas/nutrient exchange (syncytiotrophoblast/STB) and invasion and maternal vascular remodeling (extravillous trophoblast/EVT). Study of early human placental development is severely hampered by lack of a representative trophoblast stem cell (TSC) model, with the capacity for self-renewal and the ability to differentiate into both STB and EVT. We describe a reproducible protocol, using defined media containing BMP4, by which human embryonic stem cells (hESC) can be differentiated into CDX2+/p63+ CTB-like cells. These cells can be replated to further differentiate into STB- and EVT-like cells, based on marker expression, hormone secretion and invasive ability. Differentiation of hPSC-derived CTB in hypoxia leads to reduced hCG secretion and STB-associated gene expression, instead inducing EVT differentiation in a hypoxia-inducible factor-dependent manner.
2016-06-16 | GSE76090 | GEO
Project description:The phenotype of term, human placental extravillous trophoblast (EVT) reflects both the first trimester differentiation from villous cytotrophoblast (CTB) and later gestational changes, including the loss of proliferative and invasive capacity. Invasion abnormalities, as observed in preeclampsia and placenta accreta spectrum, are not usually diagnosed until the second or even third trimester of pregnancy. Characterization of the normal processes at term, including arrest of invasion is therefore crucial. In this report gene expression analysis demonstrates definitively the epithelial-mesenchymal transition (EMT) mechanism which underlies differentiation and provides a trophoblast-specific EMT signature. Methylation profiling shows that CTB, already hypomethylated relative to other somatic cells, show a further degree of hypomethylation in their transition to EVT. A small fraction of genes show both gain of methylation and changes in gene expression. Prominent are genes involved in the EMT such as the transcription factor RUNX1, loss of which leads to reduced migratory capacity in JEG3 trophoblast cells. Examination of these EMT genes leads us to suggest that the gains of methylation may assist in maintaining term EVT in a mesenchymal but non-invasive state.
2024-07-03 | GSE259304 | GEO
Project description:The phenotype of term, human placental extravillous trophoblast (EVT) reflects both the first trimester differentiation from villous cytotrophoblast (CTB) and later gestational changes, including the loss of proliferative and invasive capacity. Invasion abnormalities, as observed in preeclampsia and placenta accreta spectrum, are not usually diagnosed until the second or even third trimester of pregnancy. Characterization of the normal processes at term, including arrest of invasion is therefore crucial. In this report gene expression analysis demonstrates definitively the epithelial-mesenchymal transition (EMT) mechanism which underlies differentiation and provides a trophoblast-specific EMT signature. Methylation profiling shows that CTB, already hypomethylated relative to other somatic cells, show a further degree of hypomethylation in their transition to EVT. A small fraction of genes show both gain of methylation and changes in gene expression. Prominent are genes involved in the EMT such as the transcription factor RUNX1, loss of which leads to reduced migratory capacity in JEG3 trophoblast cells. Examination of these EMT genes leads us to suggest that the gains of methylation may assist in maintaining term EVT in a mesenchymal but non-invasive state.
2024-07-03 | GSE256412 | GEO
Project description:During pregnancy, trophoblast cells in the placenta are the only fetal cells in direct contact with the maternal blood and decidua. They have many functions, including transport of nutrients and oxygen, remodelling the uterine arteries, and communicating with maternal cells. Despite their importance in development and in the success of pregnancy, little is known about human trophoblast progenitors and their differentiation. We identify a proliferative trophoblast niche at the base of cytotrophoblast cell columns in first trimester placentas that is characterised by integrin α2 (ITGA2) expression. Pulse-chase experiments with 5-Iodo-2′-deoxyuridine (IdU) imply that these cells can contribute to both villous and extravillous lineages. Importantly, these cells can be isolated by ITGA2 using flow cytometry and express genes from both VCT and EVT. Microarray shows that they display a unique gene signature including NOTCH signalling and mesenchymal characteristics. ITGA2 allows for the first time the study of a pure population of trophoblast progenitor cells.
2018-03-18 | GSE106852 | GEO
Project description:The goal of this study was to identify the gene expression signatures of two closely related types of trophoblast in human placentas: villous cytotrophoblasts (vCTB) and proximal column extravillous trophoblasts (pcEVT). The two populations were isolated from first trimester placentas and identified using the specific surface markers, EGFR (vCTB) and HLA-G (pcEVT).
2017-02-01 | GSE80996 | GEO