Defective decidualization during and after severe preeclampsia reveals a possible maternal contribution to the etiology

In preeclampsia (PE), cytotrophoblast (CTB) invasion of the uterus and spiral arteries is often shallow. Thus, the placenta’s role has been a focus. In this study, we tested the hypothesis that decidual defects are an important determinant of the placental phenotype. We isolated human endometrial stromal cells from nonpregnant donors with a previous pregnancy that was complicated by severe PE (sPE). Compared with control cells, they failed to decidualize in vitro as demonstrated by morphological criteria and the analysis of stage-specific antigens (i.e., IGFBP1, PRL). These results were bolstered by global transcriptional profiling data that showed they were transcriptionally inert. Additionally, we used laser microdissection to isolate the decidua from tissue sections of the maternal–fetal interface in sPE. Global transcriptional profiling revealed defects in gene expression. Also, decidual cells from patients with sPE, which dedifferentiated in vitro, failed to redecidualize in culture. Conditioned medium from these cells failed to support CTB invasion. To mimic aspects of the uterine environment in normal pregnancy, we added PRL and IGFBP1, which enhanced invasion. These data suggested that failed decidualization is an important contributor to down-regulated CTB invasion in sPE. Future studies will be aimed at determining whether this discovery has translational potential with regard to assessing a woman’s risk of developing this pregnancy complication.

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Norepinephrine exposure restrains endometrial decidualization in early pregnancy

Journal of Endocrinology ◽

10.1530/joe-20-0479 ◽

2021 ◽

Author(s):

Jiju Wang ◽

Yuhui Tang ◽

Songcun Wang ◽

Liyuan Cui ◽

Da-Jin Li ◽

...

Keyword(s):

Stromal Cells ◽

Adrenergic Receptor ◽

Enrichment Analysis ◽

Normal Pregnancy ◽

Receptor Expression ◽

Gene Set Enrichment Analysis ◽

Endometrial Stromal Cells ◽

Significant Enrichment

Previous studies have focused on the role of norepinephrine on arrhythmias, generalized anxiety disorder, and cancer. This study aimed to investigate the effect of norepinephrine on endometrial decidualization. Artificial decidualization and norepinephrine-treated mice were established in vivo. In vitro, human endometrial stromal cells were treated with MPA and cAMP to induce decidualization. Decidual markers and important signaling molecules during decidualization were detected using quantitative real-time polymerase chain reaction and Western blot. RNA sequencing was performed to determine related signaling pathways. Exposure of excess norepinephrine significantly restricted the induced expression of decidualized markers Dtprp, BMP2, WNT4, and Hand2 in mice. In vitro, 10 µM norepinephrine markedly downregulated the expressions of prolactin, IGFBP1, and PLZF, which are the specifical markers of decidual stromal cells during decidualization. The gene set enrichment analysis showed that a significant enrichment in neuroactive ligand–receptor interactions of norepinephrine treatment group. The α1b-adrenergic receptor expression was upregulated by norepinephrine. Interestingly, norepinephrine did not inhibit the expression of IGFBP1 in endometrial stromal cells after silencing α1b-adrenergic receptor, while significantly suppressed the induced decidualization with overexpression of α1b-adrenergic receptor. When α1b-adrenergic receptor was activated, endometrial p-PKC was significantly increased under post-treatment with norepinephrine in vivo and in vitro. In addition, norepinephrine treatment inhibited embryo and fetal development using a normal pregnancy model. Therefore, norepinephrine exposure inhibited endometrial decidualization through the activation of the PKC signaling pathway by upregulating α1b-adrenergic receptor. Our study could explain some female reproductive problems due to stress and provide some novel strategies for this disorder.

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Expression of Wilms’ Tumor Suppressor Gene (WT1) in Human Endometrium: Regulation through Decidual Differentiation

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jcem.86.12.8074 ◽

2001 ◽

Vol 86 (12) ◽

pp. 5964-5972

Author(s):

Antonis Makrigiannakis ◽

George Coukos ◽

Anastasia Mantani ◽

Prokopis Prokopakis ◽

Geoffrey Trew ◽

...

Keyword(s):

Menstrual Cycle ◽

Stromal Cells ◽

Wilms Tumor ◽

Suppressor Gene ◽

Secretory Phase ◽

Endometrial Stromal Cells ◽

Protein Levels ◽

Decidual Cells ◽

Wt1 Protein

The Wilms’ tumor suppressor gene (WT1) encodes a zinc-finger containing transcription factor that is selectively expressed in the developing urogenital tract and functions as a tissue-specific developmental regulator. In addition to its gene-regulatory function through DNA binding properties, WT-1 also regulates transcription by formation of protein-protein complexes. These properties place WT-1 as a major regulator of cell growth and differentiation. In view of these observations, we studied WT1 mRNA and protein in human endometrial extracts and in endometrial stromal cells (ESCs) differentiating into decidual cells in vitro, by RT-PCR and Western blotting, respectively. WT1 protein expression was also studied in situ in the proliferative and the secretory phase of the menstrual cycle in the early pregnant state. Analysis by PCR of total RNA prepared from human ESCs demonstrated the presence of WT1 mRNA and four WT1 mRNA splice variants. Western blot analysis of nuclear protein extracts from ESCs yielded one immunoreactive protein of the expected size (approximately 52–54 kDa) recognized by the WT1 antibody. Immunohistochemical staining showed that WT1 protein is localized only to nuclei of human endometrial stromal cells. It remains constant in the proliferative and the secretory phase of the menstrual cycle and is increased remarkably during decidualization in early pregnancy. ESCs decidualized in vitro were investigated for WT-1 expression, which confirmed that decidualizing stimuli (E2, medroxy-progesterone-acetate, and relaxin for 12 d or cAMP and progesterone for 1–4 d) induced WT-1 mRNA (P < 0.05) and increased protein levels (P < 0.05). These data indicate that in humans the WT1 gene is expressed in ESCs and its mRNA and protein levels remain constant in the proliferative and the secretory phase of the menstrual cycle and that WT1 mRNA and protein expression increases significantly in ESCs when these cells differentiate into decidual cells.

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Thyroid Hormone Facilitates in vitro Decidualization of Human Endometrial Stromal Cells via Thyroid Hormone Receptors

Endocrinology ◽

10.1210/endocr/bqaa049 ◽

2020 ◽

Vol 161 (6) ◽

Cited By ~ 2

Author(s):

Maiko Kakita-Kobayashi ◽

Hiromi Murata ◽

Akemi Nishigaki ◽

Yoshiko Hashimoto ◽

Shinnosuke Komiya ◽

...

Keyword(s):

Thyroid Hormone ◽

Stromal Cells ◽

Hormone Receptor ◽

Expression Patterns ◽

Combined Treatment ◽

Ovarian Steroid ◽

Endometrial Stromal Cells ◽

Decidual Cells ◽

The Impact

Abstract Endometrial stromal cells differentiate into decidual cells through the process of decidualization. This differentiation is critical for embryo implantation and the successful establishment of pregnancy. Recent epidemiological studies have suggested that thyroid hormone is important in the endometrium during implantation, and it is commonly believed that thyroid hormone is essential for proper development, differentiation, growth, and metabolism. This study aimed to investigate the impact of thyroid hormone on decidualization in human endometrial stromal cells (hESCs) and define its physiological roles in vitro by gene targeting. To identify the expression patterns of thyroid hormone, we performed gene expression profiling of hESCs during decidualization after treating them with the thyroid hormone levothyroxine (LT4). A major increase in decidual response was observed after combined treatment with ovarian steroid hormones and thyroid hormone. Moreover, LT4 treatment also affected the regulation of many transcription factors important for decidualization. We found that type 3 deiodinase, which is particularly important in fetal and placental tissues, was upregulated during decidualization in the presence of thyroid hormone. Further, it was observed that progesterone receptor, an ovarian steroid hormone receptor, was involved in thyroid hormone–induced decidualization. In the absence of thyroid hormone receptor (TR), due to the simultaneous silencing of TRα and TRβ, thyroid hormone expression was unchanged during decidualization. In summary, we demonstrated that thyroid hormone is essential for decidualization in the endometrium. This is the first in vitro study to find impaired decidualization as a possible cause of infertility in subclinical hypothyroidism (SCH) patients.

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211. Proteomic identification of caldesmon as one of the physiological substrates of proprotein convertase 6 during decidualisation

Reproduction Fertility and Development ◽

10.1071/srb08abs211 ◽

2008 ◽

Vol 20 (9) ◽

pp. 11

Author(s):

B. M. Hardman ◽

L. M. Kilpatrick ◽

A. N. Stephens ◽

J. I. C. Chen ◽

P. Stanton ◽

...

Keyword(s):

Stromal Cells ◽

Hypoxia Inducible Factor ◽

Immunohistochemical Analysis ◽

Spectrometric Analysis ◽

Structural Protein ◽

Proprotein Convertase ◽

Endometrial Stromal Cells ◽

Decidual Cells

We have previously demonstrated that proprotein convertase 5/6 (PC6), a member of the proprotein convertase (PC) family, is a critical endometrial factor for implantation. PC6 is upregulated in the endometrium specifically at implantation in association with epithelial differentiation (in human and monkey) and stromal cell decidualisation (in the mouse, human and monkey). Knockdown of endometrial PC6 during early pregnancy in mice in vivo led to complete failure of implantation, while blocking of PC6 production in human endometrial stromal cells in vitro inhibited decidualisation. PCs convert a range of precursor proteins of important functions into their bioactive forms; they are thus regarded as critical ‘master switch’ molecules. We hypothesise that PC6 exerts its roles in the endometrium by regulating proteins of diverse functions essential for implantation. In this study, we utilised proteomic technology and aimed to identify proteins that are specifically cleaved by PC6 in human endometrial stromal cells (HESC) during decidualisation. HESC were decidualised with cyclic AMP, the cell lysates were treated with and without recombinant human PC6-A (rPC6-A), and the 2D Differential in Gel Electrophoresis (2D DiGE) protein profiles were compared between the two treatments. We identified several proteins which were differentially cleaved following the addition of rPC6-A. Mass spectrometric analysis confirmed that the most abundant of these were caldesmon, tropomyosin-2, tropomyosin-4, hypoxia Inducible factor-1 and chloride intracellular channel-1. These proteins showed spot shifts in hPC6-A treated HESC lysates consistent with hPC6-A cleavage. western blot analysis confirmed the specific cleavage of caldesmon by PC6 in HESCs, and immunohistochemical analysis showed co-localisation of caldesmon and PC6 in decidual cells in human endometrial tissue. Given that caldesmon is a structural protein previously found to be involved in actin filament reorganisation, our results strongly suggest that PC6 is a mediator of structural remodelling of stromal cells during decidualisation in the endometrium.

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Dehydroepiandrosterone Inhibits Glucose Flux Through the Pentose Phosphate Pathway in Human and Mouse Endometrial Stromal Cells, Preventing Decidualization and Implantation

Molecular Endocrinology ◽

10.1210/me.2011-0026 ◽

2011 ◽

Vol 25 (8) ◽

pp. 1444-1455 ◽

Cited By ~ 44

Author(s):

Antonina I. Frolova ◽

Kathleen O'Neill ◽

Kelle H. Moley

Keyword(s):

Stromal Cells ◽

Pentose Phosphate Pathway ◽

Glucose Transporter ◽

Polycystic Ovary ◽

Pentose Phosphate ◽

Reproductive Age ◽

Endometrial Stromal Cells ◽

Glucose Transporter 1 ◽

Decidual Cells

Endometrial stromal cells (ESC) must undergo a hormone-driven differentiation to form decidual cells as a requirement of proper embryo implantation. Recent studies from our laboratory have demonstrated that decidualizing cells require glucose transporter 1 expression and an increase in glucose use to complete this step. The present study focuses on the glucose-dependent molecular and metabolic pathways, which are required by ESC for decidualization. Inhibition of glycolysis had no effect on decidualization. However, blockade of the pentose phosphate pathway (PPP) with pharmacologic inhibitors 6-aminonicotinamide or dehydroepiandrosterone (DHEA), and short hairpin RNA-mediated knockdown of glucose-6-phosphate dehydrogenase, the rate-limiting step in the PPP, both led to strong decreases in decidual marker expression in vitro and decreased decidualization in vivo. Additionally, the studies demonstrate that inhibition is due, at least in part, to ribose-5-phosphate depletion, because exogenous nucleoside administration restored decidualization in these cells. The finding that PPP inhibition prevents decidualization of ESC is novel and clinically important, because DHEA is an endogenous hormone produced by the adrenal glands and elevated in a high proportion of women who have polycystic ovary syndrome, the most common endocrinopathy in reproductive age women. Together, this data suggest a mechanistic link between increased DHEA levels, use of glucose via the PPP, and pregnancy loss.

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Deciphering the role of PGRMC1 during human decidualization using an in vitro approach

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/clinem/dgab303 ◽

2021 ◽

Author(s):

Stefania Salsano ◽

Roberto González-Martín ◽

Alicia Quiñonero ◽

Silvia Pérez-Debén ◽

Francisco Domínguez

Keyword(s):

Progesterone Receptor ◽

Intracellular Transport ◽

Transcriptional Profiling ◽

Molecular Transport ◽

In Silico Analysis ◽

Prolactin Secretion ◽

Mitochondrial Activity ◽

Endometrial Stromal Cells ◽

Membrane Progesterone Receptor

Abstract Context Non-classical membrane progesterone receptor (mPRs) and PGRMC1 expression have been detected in endometrium, but their role in decidualization was not yet investigated. We previously demonstrated PGRMC1 downregulation in receptive endometrium and that its overexpression inhibits decidualization. Furthermore, during decidualization, PGRMC1 mainly interacts with proteins involved in biosynthesis, intracellular transport and mitochondrial activity. Objective To determine PGRMC1 and mPRs signaling role during decidualization. Design and Interventions Isolated primary endometrial stromal cells (EnSC) were in vitro decidualized in presence of classic stimuli (E2+P4), PGRMC1 inhibitor (AG205), or membrane-impermeable P4 (P4-BSA). Setting and Participants Endometrial biopsies from 19 fertile oocyte donors attending IVI-Valencia IVF clinic. Main Outcome Measure(s) EnSC decidualization was evaluated by prolactin ELISA and F-actin immunostaining. Progesterone receptor localization was evaluated by immunofluorescence. EnSC transcriptomic profiles were analyzed by microarray technology. Result(s) PGRMC1 inhibition during EnSC decidualization (AG205dEnSC) does not interfere with EnSC cytoskeletal rearrangements and prolactin secretion. However, global transcriptional profiling revealed more differentially expressed genes in AG205dEnSC than in dEnSC, compared with non-decidualized EnSC (ndEnSC). In silico analysis showed that PGRMC1 inhibition upregulated more genes related to metabolism, molecular transport, and hormonal biosynthesis compared to control dEnSC. EnSC decidualized in the presence of P4-BSA showed a similar behavior as ndEnSC in terms of morphological features, absence of prolactin secretion, and transcriptomic pattern. Conclusion(s) Our findings associate PGRMC1 to hormonal biosynthesis, metabolism, and vesicular transport—important cellular functions for dEnSC supporting pregnancy. Activation of membrane P4 receptor signaling alone was unable to induce downstream effects needed for proper decidualization.

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Bisphenol A modulates receptivity and secretory function of human decidual cells: an in vitro study

Reproduction ◽

10.1530/rep-14-0601 ◽

2015 ◽

Vol 150 (2) ◽

pp. 115-125 ◽

Cited By ~ 19

Author(s):

Chiara Mannelli ◽

Anna Z Szóstek ◽

Karolina Lukasik ◽

Claudiopietro Carotenuto ◽

Francesca Ietta ◽

...

Keyword(s):

Bisphenol A ◽

Cell Viability ◽

Hormone Receptors ◽

Progesterone Receptors ◽

In Vitro Study ◽

Endometrial Stromal Cells ◽

Decidual Cells ◽

Decidual Prolactin ◽

Toxic Concentrations

The human endometrium is a fertility-determining tissue and a target of steroid hormones' action. Endocrine disruptors (EDs) can exert adverse effects on the physiological function of the decidua at the maternal–fetal interface. We examined the potential effects of an ED, bisphenol A (BPA), on endometrial maturation/decidualization, receptivity, and secretion of decidual factors (biomarkers). In vitro decidualized, endometrial stromal cells from six hysterectomy specimens were treated with 1 pM–1 μM of BPA, for 24 h and assessed for cell viability and proliferation. Three non-toxic concentrations of BPA (1 μM, 1 nM, and 1 pM) were selected to study its influence on secretion of cell decidualization biomarkers (IGF-binding protein and decidual prolactin (dPRL)), macrophage migration inhibitory factor (MIF) secretion, and hormone receptors' expression (estrogen receptors (ERα and ERβ); progesterone receptors (PRA and PRB); and human chorionic gonadotropin (hCG)/LH receptor (LH-R)). The results showed a decrease in cell viability (P<0.001) in response to BPA at the level of 1 mM. At the non-toxic concentrations used, BPA perturbed the expression of ERα, ERβ, PRA, PRB, and hCG/LH-R (P<0.05). Furthermore, 1 μM of BPA reduced the mRNA transcription of dPRL (P<0.05). Secretion of MIF was stimulated by all BPA treatments, the lowest concentration (1 pM) being the most effective (P<0.001). The multi-targeted disruption of BPA on decidual cells, at concentrations commonly detected in the human population, raises great concern about the possible consequences of exposure to BPA on the function of decidua and thus its potential deleterious effect on pregnancy.

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Pregnancy and interferon τ regulate DDX58 and PLSCR1 in the ovine uterus during the peri-implantation period

Reproduction ◽

10.1530/rep-10-0348 ◽

2011 ◽

Vol 141 (1) ◽

pp. 127-138 ◽

Cited By ~ 12

Author(s):

Gwonhwa Song ◽

Jo-Ann G W Fleming ◽

Jinyoung Kim ◽

Thomas E Spencer ◽

Fuller W Bazer

Keyword(s):

Biological Effects ◽

Transcriptional Profiling ◽

Type I ◽

Null Cell ◽

Endometrial Stromal Cells ◽

Endometrial Cells ◽

Phospholipid Scramblase ◽

Steady State Levels ◽

Ovine Uterus

Interferon τ (IFNT), the pregnancy recognition signal in ruminants, abrogates the luteolytic mechanism for maintenance of the corpus luteum for production of progesterone (P4). This study examined the expression of DEAD (Asp-Glu-Ala-Asp) box polypeptide 58 (DDX58) and phospholipid scramblase 1 (PLSCR1) mRNAs in the ovine uterus as these genes were increased most in 2fTGH (STAT1 positive) cells by IFNT. The results of this study indicated that IFNT regulates expression ofDDX58andPLSCR1mRNAs in the ovine uterus, which confirmed the results of thein vitrotranscriptional profiling experiment with the 2fTGH (parental STAT1 positive) and U3A (STAT1 null) cell lines. Steady-state levels ofDDX58andPLSCR1mRNAs increased in cells of the ovine uterus between days 12 and 20 of pregnancy, but not between days 10 and 16 of the estrous cycle. The expression ofDDX58andPLSCR1mRNAs was greatest in endometrial stromal cells, but there was transient expression in uterine luminal and superficial glandular epithelial cells. P4alone did not induce expression ofDDX58andPLSCR1mRNAs; however, intrauterine injections of IFNT did induce expression ofDDX58andPLSCR1mRNAs in the endometria of nonpregnant ewes independent of effects of P4. These results indicate that IFNT induces expression ofDDX58andPLSCR1in ovine endometrial cells via the classical STAT1-mediated cell signaling pathway. Based on their known biological effects,DDX58andPLSCR1are IFN-stimulated genes, which may increase the antiviral status of cells of the pregnant uterus to protect against viral infection and/or enhance secretion of type I IFNs that inhibit viral replication.

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Hsa-miR-222 Is Involved in Differentiation of Endometrial Stromal Cells in Vitro

Endocrinology ◽

10.1210/en.2008-1629 ◽

2009 ◽

Vol 150 (10) ◽

pp. 4734-4743 ◽

Cited By ~ 52

Author(s):

Kun Qian ◽

Linli Hu ◽

Hong Chen ◽

Haixia Li ◽

Na Liu ◽

...

Keyword(s):

Stromal Cells ◽

Target Genes ◽

Embryo Implantation ◽

Endometrial Stromal Cells ◽

Protein Levels ◽

Mirna Genes ◽

False Discovery ◽

Decidual Cells ◽

Expression Variance

Abstract Decidualization is a critical step during embryo implantation and characterized by the differentiation of endometrial stromal cells (ESCs) into decidual cells. Because miRNAs are important determinants of cellular fate specification, in this study, the miRNA expression in ESCs during in vitro decidualization was profiled by using a microarray. Significance analysis of microarrays revealed that 49 miRNA genes were differently (>2-fold) expressed between the noninduced ESCs and induced ESCs with a false discovery rate of 0. The expression variance of hsa-miR-222, 221, 143, 101, 30d, 30c, 181b, 27b, 29b, 507, and 23a was validated by using quantitative PCR (P < 0.05). Based on microRNA (miRNA) and mRNA expression variance and predicted target genes of miRNAs, a bioinformatic model of miRNAs controlling ESCs differentiation was formulated. Finally, we proved that down-regulation of has-miR-222 could decrease the number of cells in S phase during ESCs differentiation (P < 0.05). Antisense oligonucleotides of has-miR-222 could increase reporter gene expression by targeting the 3′ untranslated regions of CDKN1C/p57kip2 mRNAs as well as increase CDKN1C/p57kip2 protein levels (P < 0.05). In conclusion, our results suggest that a subset of miRNAs play a key role in gene reprogramming during ESCs decidualization and that hsa-miR-222 participates in ESC differentiation by regulating ESCs terminally withdrawing from the cell cycle.

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Notch1 Is Regulated by Chorionic Gonadotropin and Progesterone in Endometrial Stromal Cells and Modulates Decidualization in Primates

Endocrinology ◽

10.1210/en.2011-2122 ◽

2012 ◽

Vol 153 (6) ◽

pp. 2884-2896 ◽

Cited By ~ 44

Author(s):

Yalda Afshar ◽

Lucio Miele ◽

Asgerally T. Fazleabas

Keyword(s):

Chorionic Gonadotropin ◽

Cellular Proliferation ◽

The Body ◽

Stromal Fibroblast ◽

Uterine Receptivity ◽

Endometrial Stromal Cells ◽

Decidual Cells ◽

Short Period ◽

Extensive Growth

No other tissue in the body undergoes such a vast and extensive growth and remodeling in a relatively short period of time as the primate endometrium. Endometrial integrity is coordinated by ovarian hormones, namely, estrogens, progesterone, and the embryonic hormone chorionic gonadotropin (CG). These regulated events modulate the menstrual cycle and decidualization. The Notch family of transmembrane receptors regulate cellular proliferation, differentiation, and apoptosis, cellular processes required to maintain endometrial integrity. In two primate models, the human and the simulated pregnant baboon model, we demonstrated that Notch1 is increased during the window of uterine receptivity, concomitant with CG. Furthermore, CG combined with estrogens and progesterone up-regulate the level of Notch1, whereas progesterone increases the intracellular transcriptionally competent Notch1, which binds in a complex with progesterone receptor. Inhibition of Notch1 prevented decidualization, and alternatively, when decidualization is biochemically recapitulated in vitro, Notch1 is down-regulated. A focused microarray demonstrated that the Notch inhibitor, Numb, dramatically increased when Notch1 decreased during decidualization. We propose that in the endometrium, Notch has a dual role during the window of uterine receptivity. Initially, Notch1 mediates a survival signal in the uterine endometrium in response to CG from the implanting blastocyst and progesterone, so that menstrual sloughing is averted. Subsequently, Notch1 down-regulation may be critical for the transition of stromal fibroblast to decidual cells, which is essential for the establishment of a successful pregnancy.

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