Dissecting the First Transcriptional Divergence During Human Embryonic Development

Qiang, Boqin; Assou, Saïd; Haouzi, D.; Ramirez, Jean-Marie; Monzo, C.; Becker, Fabienne; Gerbal‐Chaloin, Sabine; Hamamah, S.; Vos, John De

doi:10.1007/s12015-011-9301-3

Cited by 75 publications

(81 citation statements)

References 59 publications

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“…Gene expression analysis of a previously published list of human TE-specific transcription factors showed induction of 12 out of 16 TE-specific transcription factors over the first 6 days of differentiation (Fig. 1E) (Bai et al, 2012). These included induction of TP63 (further discussed below) and PPARG; the latter has been identified by our own studies and those of others as an upstream regulator of the GCM1-ERVW1-directed pathway of syncytiotrophoblast differentiation (Baczyk et al, 2009;Parast et al, 2009).…”

Section: Comparative Gene Expression Profiling Of Human Fetal and Plamentioning

confidence: 53%

“…Trophoblast-associated genes were selected based on previously published studies, with trophectoderm (TE)-specific genes inferred specifically from Bai et al (Bai et al, 2012). Cut-offs were set to variance=0.02 and multi-group comparison q≤0.01.…”

Section: Analysis Of Gene Expression Profiling Datamentioning

confidence: 99%

“…Over the past decade, multiple studies have identified human pluripotent stem cells (hPSCs), including embryonic (hESCs) and induced pluripotent stem cells (hiPSCs), as useful models for trophoblast differentiation; specifically, in the presence of feederconditioned medium (FCM), BMP4 induces this lineage, with production of both human chorionic gonadotrophin (hCG)β-secreting STB and surface HLA-G-expressing EVT (Xu et al, 2002;Das et al, 2007;Wu et al, 2008;Marchand et al, 2011;Bai et al, 2012). These results imply the presence, however transient, of a trophoblast progenitor population; in fact, a recent report indicates that surface aminopeptidase A (APA)-positive cells are progenitors, at least for multinucleated STB (Drukker et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

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BMP4-directed trophoblast differentiation of human embryonic stem cells is mediated through a ΔNp63+ cytotrophoblast stem cell state

et al. 2013

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SUMMARYThe placenta is a transient organ that is necessary for proper fetal development. Its main functional component is the trophoblast, which is derived from extra-embryonic ectoderm. Little is known about early trophoblast differentiation in the human embryo, owing to lack of a proper in vitro model system. Human embryonic stem cells (hESCs) differentiate into functional trophoblast following BMP4 treatment in the presence of feeder-conditioned media; however, this model has not been widely accepted, in part owing to a lack of proof for a trophoblast progenitor population. We have previously shown that p63, a member of the p53 family of nuclear proteins, is expressed in proliferative cytotrophoblast (CTB), precursors to terminally differentiated syncytiotrophoblast (STB) in chorionic villi and extravillous trophoblast (EVT) at the implantation site. Here, we show that BMP4-treated hESCs differentiate into bona fide CTB by direct comparison with primary human placental tissues and isolated CTB through gene expression profiling. We show that, in primary CTB, p63 levels are reduced as cells differentiate into STB, and that forced expression of p63 maintains cyclin B1 and inhibits STB differentiation. We also establish that, similar to in vivo events, hESC differentiation into trophoblast is characterized by a p63 + /KRT7 + CTB stem cell state, followed by formation of functional KLF4 + STB and HLA-G + EVT. Finally, we illustrate that downregulation of p63 by shRNA inhibits differentiation of hESCs into functional trophoblast. Taken together, our results establish that BMP4-treated hESCs are an excellent model of human trophoblast differentiation, closely mimicking the in vivo progression from p63 + CTB stem cells to terminally differentiated trophoblast subtypes.

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Section: Comparative Gene Expression Profiling Of Human Fetal and Plamentioning

confidence: 53%

Section: Analysis Of Gene Expression Profiling Datamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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BMP4-directed trophoblast differentiation of human embryonic stem cells is mediated through a ΔNp63+ cytotrophoblast stem cell state

et al. 2013

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“…In this study we confirm that the early human placenta is composed of a population of CDX2 + -p63 + double-positive CTBs, which we propose as the CTB "stem" cell compartment. p63 has been shown to be enriched, at least at the RNA level, in even earlier stages of human trophoblast, specifically, in the TE of human blastocysts (45). Therefore it is possible that this CDX2 + -p63 + double-positive population may emerge in the human blastocyst and persist in a subset of trophoblasts in the early postimplantation placenta.…”

Section: Discussionmentioning

confidence: 99%

Human pluripotent stem cells as a model of trophoblast differentiation in both normal development and disease

Horii

Wakeland

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

141

164

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Trophoblast is the primary epithelial cell type in the placenta, a transient organ required for proper fetal growth and development. Different trophoblast subtypes are responsible for gas/nutrient exchange (syncytiotrophoblasts, STBs) and invasion and maternal vascular remodeling (extravillous trophoblasts, EVTs). Studies of early human placental development are severely hampered by the lack of a representative trophoblast stem cell (TSC) model with the capacity for self-renewal and the ability to differentiate into both STBs and EVTs. Primary cytotrophoblasts (CTBs) isolated from early-gestation (6-8 wk) human placentas are bipotential, a phenotype that is lost with increasing gestational age. We have identified a CDX2 + /p63 + CTB subpopulation in the early postimplantation human placenta that is significantly reduced later in gestation. We describe a reproducible protocol, using defined medium containing bone morphogenetic protein 4 by which human pluripotent stem cells (hPSCs) can be differentiated into CDX2 + /p63 + CTB stem-like cells. These cells can be replated and further differentiated into STB-and EVT-like cells, based on marker expression, hormone secretion, and invasive ability. As in primary CTBs, differentiation of hPSC-derived CTBs in low oxygen leads to reduced human chorionic gonadotropin secretion and STB-associated gene expression, instead promoting differentiation into HLA-G + EVTs in an hypoxiainducible, factor-dependent manner. To validate further the utility of hPSC-derived CTBs, we demonstrated that differentiation of trisomy 21 (T21) hPSCs recapitulates the delayed CTB maturation and blunted STB differentiation seen in T21 placentae. Collectively, our data suggest that hPSCs are a valuable model of human placental development, enabling us to recapitulate processes that result in both normal and diseased pregnancies.human pluripotent stem cells | cytotrophoblast | extravillous trophoblast | syncytiotrophoblast | placenta

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“…Of 63 trophoblast gene markers examined (18,21), only one, CDH1 (encoding E-cadherin), was not up-regulated in the ESC >70-fraction compared with the ESCu (Fig. 5A).…”

Section: Isolation Of Syncytial Areas From Colonies Of Bap-treated H1mentioning

confidence: 98%

Comparison of syncytiotrophoblast generated from human embryonic stem cells and from term placentas

Yabe

Alexenko

Amita

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

149

148

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Human embryonic stem cells (ESCs) readily commit to the trophoblast lineage after exposure to bone morphogenetic protein-4 (BMP-4) and two small compounds, an activin A signaling inhibitor and a FGF2 signaling inhibitor (BMP4/A83-01/PD173074; BAP treatment). During differentiation, areas emerge within the colonies with the biochemical and morphological features of syncytiotrophoblast (STB). Relatively pure fractions of mononucleated cytotrophoblast (CTB) and larger syncytial sheets displaying the expected markers of STB can be obtained by differential filtration of dispersed colonies through nylon strainers. RNA-seq analysis of these fractions has allowed them to be compared with cytotrophoblasts isolated from term placentas before and after such cells had formed syncytia. Although it is clear from extensive gene marker analysis that both ESC-and placenta-derived syncytial cells are trophoblast, each with the potential to transport a wide range of solutes and synthesize placental hormones, their transcriptome profiles are sufficiently dissimilar to suggest that the two cell types have distinct pedigrees and represent functionally different kinds of STB. We propose that the STB generated from human ESCs represents the primitive syncytium encountered in early pregnancy soon after the human trophoblast invades into the uterine wall.is encountered during at least two stages of human placental development (1-3). The first coincides with early implantation when a multinucleated syncytium forms, presumably by cell fusion events, ahead of proliferating, mononucleated, cytotrophoblast (CTB) cells originating from polar trophectoderm (3,4). This invasive syncytium emerges either during or soon after the trophoblast passes through the breached uterine epithelium and into the decidualized stromal layer beneath and appears to be responsible for hollowing out regions within the stroma to form lacunae (5), which become filled with fluid and cells from maternal blood and uterine glands and presumably provide a source of nutrients for the conceptus (3, 6). By about 12 d of gestation, soon after the blastocyst has sunk below the endometrial surface, strands of cytotrophoblast begin to form and penetrate through the primitive syncytium to form primary chorionic villi, which are subsequently invaded by extraembryonic mesoderm to form secondary and tertiary villi (villous trees) (2-4). The cytotrophoblast cells associated with the villi continue to divide and provide a progenitor cell population for the villous STB, which is the cell layer that covers the outer surface of the villi and forms the definitive interface involved in exchange of gases, nutrients, and excretory materials between the fetal placenta and maternal blood. Villous STB is also the major site for production of placental hormones. Cytotrophoblast cells at the tips of the anchoring villi proliferate and colonize the endometrium, thus expanding the placental bed and simultaneously remodeling maternal spiral arteries. The extent to which extravillous trophoblast becom...

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Dissecting the First Transcriptional Divergence During Human Embryonic Development

Cited by 75 publications

References 59 publications

BMP4-directed trophoblast differentiation of human embryonic stem cells is mediated through a ΔNp63+ cytotrophoblast stem cell state

BMP4-directed trophoblast differentiation of human embryonic stem cells is mediated through a ΔNp63+ cytotrophoblast stem cell state

Human pluripotent stem cells as a model of trophoblast differentiation in both normal development and disease

Comparison of syncytiotrophoblast generated from human embryonic stem cells and from term placentas

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