Differentiation of first trimester cytotrophoblast to extravillous trophoblast involves an epithelial–mesenchymal transition

DaSilva-Arnold, Sonia; James, Joanna L.; Al-Khan, Abdulla; Zamudio, Stacy; Illsley, Nicholas P.

doi:10.1016/j.placenta.2015.10.013

Cited by 141 publications

(96 citation statements)

References 41 publications

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“…These results indicated the effect of complete pECM on cytotrophoblast invasion cannot be reproduced using individual fractions alone, suggesting a combination mechanism of action of both placental BM proteins and peptide components. To confirm that invasion required the expression of MMP, a known group of enzymes that are responsible for the degradation of ECM proteins and are upregulated during EMT and invasion of cytotrophoblasts, we tested the effect of fractionated and complete pECM on the gene expressions of MMP2 and MMP9 (Fig. ) .…”

Section: Resultsmentioning

confidence: 99%

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Placental basement membrane proteins are required for effective cytotrophoblast invasion in a three‐dimensional bioprinted placenta model

Kuo

Guo

Cabrera-Luque

et al. 2018

J Biomedical Materials Res

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Fetal cytotrophoblast invasion of maternal decidual vasculature is necessary to normal pregnancy. In preeclampsia, there is shallow invasion and abnormal remodeling of the uterine vasculature that lead to significant maternal and perinatal morbidity and mortality. The placental basement membrane (BM) proteins (e.g., laminin and collagen) has been implicated in the development of placenta while the level of laminin is significantly lower in preeclampsia. However, there are very limited studies, if any, on the effect of extracellular matrix (ECM) microenvironment on the invasion of cytotrophoblast. In this study, we hypothesized that placental BM proteins are required for effective cytotrophoblast invasion. Using proteomics, we found that more than 80% of ECM proteins in placental basal plate (pECM) were BM proteins. In addition to upregulating expressions of MMP2 (1.5-fold) and MMP9 (6.3-fold), pECM significantly increased the motility rates of cytotrophoblasts by 13-fold (from 5.60 ± 0.95 to 75.5 ± 21.8 µm/day) to achieve an effective invasion rate that was comparable to in vivo results. Treatments with PI3K inhibitors completely removed the pECM-enhanced invasive phenotypes and genotypes of cytotrophoblasts, suggesting its dominant role in cytotrophoblast-ECM interactions. Our results described, for the first time, the substantial effects of the ECM microenvironment on regulating cytotrophoblast invasion, an area that is less investigated but appear to be critical in the pathogenesis of preeclampsia. Moreover, the approach presented in this work that fabricates organ models with organ-specific ECM can be an attractive option to screen and develop novel therapeutics and biomarkers not only in preeclampsia but also other diseases such as cancer metastasis. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1476-1487, 2018.

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Section: Resultsmentioning

confidence: 99%

“…the degradation of ECM proteins and are upregulated during EMT and invasion of cytotrophoblasts, 29,45 we tested the effect of fractionated and complete pECM on the gene expressions of MMP2 and MMP9 (Fig. 5).…”

Section: Effects Of Fractionated Pecm On Cytotrophoblast Invasionmentioning

confidence: 99%

Placental basement membrane proteins are required for effective cytotrophoblast invasion in a three‐dimensional bioprinted placenta model

Kuo

Guo

Cabrera-Luque

et al. 2018

J Biomedical Materials Res

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“…Abou-Kheir et al ., showed that the HTR-8/SVneo cell line contained a heterogeneous population of trophoblast and mesenchymal/stromal cells [30]. In comparison to other placental cell lines, the expression of trophoblast/epithelial markers such as cytokeratin 7 (CK7) and e-cadherin was silenced in HTR-8/SVneo cells [30–32] while a higher expression of vimentin (a marker of epithelial to mesenchymal transition) was observed [30–33]. Similarly, Takao et al ., observed a weak expression of HLA-G and integrin alpha-V/beta-3 [34], which are known primary EVT (epithelial) markers [35–37].…”

Section: Discussionmentioning

confidence: 99%

Oxygen tension regulates the miRNA profile and bioactivity of exosomes released from extravillous trophoblast cells – Liquid biopsies for monitoring complications of pregnancy

et al. 2017

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Our understanding of how cells communicate has undergone a paradigm shift since the recent recognition of the role of exosomes in intercellular signaling. In this study, we investigated whether oxygen tension alters the exosome release and miRNA profile from extravillous trophoblast (EVT) cells, modifying their bioactivity on endothelial cells (EC). Furthermore, we have established the exosomal miRNA profile at early gestation in women who develop pre-eclampsia (PE) and spontaneous preterm birth (SPTB). HTR-8/SVneo cells were used as an EVT model. The effect of oxygen tension (i.e. 8% and 1% oxygen) on exosome release was quantified using nanocrystals (Qdot®) coupled to CD63 by fluorescence NTA. A real-time, live-cell imaging system (Incucyte™) was used to establish the effect of exosomes on EC. Plasma samples were obtained at early gestation (<18 weeks) and classified according to pregnancy outcomes. An Illumina TrueSeq Small RNA kit was used to construct a small RNA library from exosomal RNA obtained from EVT and plasma samples. The number of exosomes was significantly higher in EVT cultured under 1% compared to 8% oxygen. In total, 741 miRNA were identified in exosomes from EVT. Bioinformatic analysis revealed that these miRNA were associated with cell migration and cytokine production. Interestingly, exosomes isolated from EVT cultured at 8% oxygen increased EC migration, whilst exosomes cultured at 1% oxygen decreased EC migration. These changes were inversely proportional to TNF-α released from EC. Finally, we have identified a set of unique miRNAs in exosomes from EVT cultured at 1% oxygen and exosomes isolated from the circulation of mothers at early gestation, who later developed PE and SPTB. We suggest that aberrant exosomal signalling by placental cells is a common aetiological factor in pregnancy complications characterised by incomplete SpA remodeling and is therefore a clinically relevant biomarker of pregnancy complications.

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“…The ALK5 receptor binds TGF-ß and activin, hence the small molecule inhibitor A83-01 acts to inhibit this binding, which prevents downstream activation of SMAD signaling pathways, and altogether has been shown to inhibit the epithelial-mesenchymal transition (EMT) induced by TGF-ß signaling 71 . An EMT is thought to occur during EVT differentiation as EVTs transition from an epithelial to mesenchymal cell type with loss of their attachment to a basement membrane, allowing for migration and invasion into the maternal decidua 72,73 . DaSilva-Arnold et al 73 observed increased gene expression of TGFB1 and TGFB2 in human EVTs compared to primary villous cytotrophoblasts suggesting that TGF-ß expression is intrinsic to human EVTs.…”

Section: Moreover Macaque St-3d and Evt Cells Secreted High Levels Omentioning

confidence: 99%

Derivation of macaque trophoblast stem cells

Schmidt

Meyer

Wiepz

et al. 2020

Preprint

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200 words max) Nonhuman primates are excellent models for studying human placentation as experimental manipulations in vitro can be translated to in vivo pregnancy. Our objective was to develop macaque trophoblast stem cells (TSC) as an in vitro platform for future assessment of primate trophoblast development and function. Macaque TSC lines were generated by isolating first trimester placental villous cytotrophoblasts followed by culture in TSC medium to "reprogram" the cells to a proliferative state. TSCs grew as mononuclear colonies, whereas upon induction of syncytiotrophoblast (ST) differentiation multinuclear structures appeared, indicative of syncytium formation. Chorionic gonadotropin secretion was >4,000-fold higher in ST culture media compared to TSC media. Characteristic trophoblast hallmarks were defined in TSCs and ST including expression of C19MC miRNAs and macaque placental nonclassical MHC class I molecule, Mamu-AG. TSC differentiation to extravillous trophoblasts (EVTs) with or without the ALK-5 inhibitor A83-01 resulted in differing morphologies but similar expression of Mamu-AG and CD56 as assessed by flow cytometry, hence further refinement of relevant EVT markers is needed. Our preliminary characterization of macaque TSCs suggests that these cells represent a proliferative, self-renewing TSC population capable of differentiating to STs in vitro thereby establishing an experimental model of primate placentation.

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Differentiation of first trimester cytotrophoblast to extravillous trophoblast involves an epithelial–mesenchymal transition

Cited by 141 publications

References 41 publications

Placental basement membrane proteins are required for effective cytotrophoblast invasion in a three‐dimensional bioprinted placenta model

Placental basement membrane proteins are required for effective cytotrophoblast invasion in a three‐dimensional bioprinted placenta model

Oxygen tension regulates the miRNA profile and bioactivity of exosomes released from extravillous trophoblast cells – Liquid biopsies for monitoring complications of pregnancy

Derivation of macaque trophoblast stem cells

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