Preimplantation factor modulates trophoblastic invasion throughout the decidualization of human endometrial stromal cells

Abstract Background Successful human embryo implantation requires the differentiation of endometrial stromal cells (ESCs) into decidual cells during a process called decidualization. ESCs express specific markers of decidualization, including prolactin, insulin-like growth factor-binding protein-1 (IGFBP-1), and connexin-43. Decidual cells also control of trophoblast invasion by secreting various factors, such as matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases. Preimplantation factor (PIF) is a recently identified, embryo-derived peptide with activities at the fetal-maternal interface. It creates a favorable pro-inflammatory environment in human endometrium and directly controls placental development by increasing the human trophoblastic cells’ ability to invade the endometrium. We hypothesized that PIF’s effects on the endometrium counteract its pro-invasive effects. Methods We tested sPIF effect on the expression of three decidualization markers by RT-qPCR and/or immunochemiluminescence assay. We examined sPIF effect on human ESC migration by performing an in vitro wound healing assay. We analyzed sPIF effect on endometrial control of human trophoblast invasion by performing a zymography and an invasion assay. Results Firstly, we found that a synthetic analog of PIF (sPIF) significantly upregulates the mRNA expression of IGFBP-1 and connexin-43, and prolactin secretion in ESCs - suggesting a pro-differentiation effect. Secondly, we showed that the HTR-8/SVneo trophoblastic cell line’s invasive ability was low in the presence of conditioned media from ESCs cultured with sPIF. Thirdly, this PIF’s anti-invasive action was associated with a specifically decrease in MMP-9 activity. Conclusion Taken as a whole, our results suggest that PIF accentuates the decidualization process and the production of endometrial factors that limit trophoblast invasion. By controlling both trophoblast and endometrial cells, PIF therefore appears to be a pivotal player in the human embryo implantation process.

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Adiponectin limits differentiation and trophoblast invasion in human endometrial cells

Journal of Molecular Endocrinology ◽

10.1530/jme-17-0046 ◽

2017 ◽

Vol 59 (3) ◽

pp. 285-297 ◽

Cited By ~ 3

Author(s):

Fabien Duval ◽

Esther Dos Santos ◽

Hadia Moindjie ◽

Valérie Serazin ◽

Nelly Swierkowski-Blanchard ◽

...

Keyword(s):

Connexin 43 ◽

Embryo Implantation ◽

Tissue Inhibitors Of Metalloproteinases ◽

Trophoblast Invasion ◽

Human Escs ◽

Trophoblastic Cell ◽

Endometrial Stromal Cells ◽

Human Trophoblast ◽

Endometrial Cells ◽

Decidual Cells

Successful human embryo implantation requires a proper differentiation of endometrial stromal cells (ESCs) into decidual cells, during a process called decidualization. ESCs express specific molecules, such as prolactin, insulin-like growth factor-binding protein-1 (IGFBP-1) and connexin-43. Decidual cells are also involved in the control of trophoblast invasion, by secreting various factors, such as matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). Adiponectin is an adipokine with insulin-sensitizing, anti-inflammatory and anti-proliferative effects. At the embryo–maternal interface, adiponectin promotes differentiation and invasion of human trophoblastic cells. We hypothesize that the effects of adiponectin on endometrium could counteract its pro-invasive effects previously described in the human trophoblast. In this context, we have firstly demonstrated that adiponectin downregulates IGFBP-1 and connexin-43 mRNA expressions, as well as prolactin secretion in ESCs, suggesting an anti-differentiative effect of adiponectin. Secondly, we found that invasive capacities of trophoblastic cell line HTR-8/SVneo are reduced in the presence of conditioned media from ESC cultured in the presence of adiponectin. Adiponectin’s anti-invasive action is associated with a decreased activity of MMP-2 and MMP-9, and an increased TIMP-3 mRNA expression in ESCs. Finally, adiponectin receptors (ADIPOR1 and ADIPOR2) knockdown abolishes the anti-differentiative and anti-invasive effects of adiponectin in human ESCs. Altogether, our results suggest that adiponectin reduces the decidualization process and inversely induces the production of endometrial factors that limit trophoblast invasion. Thus, through a dual control in trophoblast and endometrial cells, adiponectin appears as a pivotal actor of the embryo implantation process.

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The autophagy protein, FIP200 (RB1CC1) mediates progesterone responses governing uterine receptivity and decidualization†

Biology of Reproduction ◽

10.1093/biolre/ioz234 ◽

2020 ◽

Vol 102 (4) ◽

pp. 843-851 ◽

Cited By ~ 2

Author(s):

Arin K Oestreich ◽

Sangappa B Chadchan ◽

Alexandra Medvedeva ◽

John P Lydon ◽

Emily S Jungheim ◽

...

Keyword(s):

Stromal Cells ◽

Embryo Implantation ◽

Conditional Knockout ◽

Trophoblast Invasion ◽

Uterine Receptivity ◽

Human Escs ◽

Embryo Survival ◽

Endometrial Stromal Cells ◽

Cellular Changes ◽

Decidual Cells

Abstract Successful establishment of pregnancy depends on steroid hormone-driven cellular changes in the uterus during the peri-implantation period. To become receptive to embryo implantation, uterine endometrial stromal cells (ESCs) must transdifferentiate into decidual cells that secrete factors necessary for embryo survival and trophoblast invasion. Autophagy is a key homeostatic process vital for cellular homeostasis. Although the uterus undergoes major cellular changes during early pregnancy, the precise role of autophagy in uterine function is unknown. Here, we report that conditional knockout of the autophagy protein FIP200 in the reproductive tract of female mice results in reduced fecundity due to an implantation defect. In the absence of FIP200, aberrant progesterone signaling results in sustained uterine epithelial proliferation and failure of stromal cells to decidualize. Additionally, loss of FIP200 impairs decidualization of human ESCs. We conclude that the autophagy protein FIP200 plays a crucial role in uterine receptivity, decidualization, and fertility. These data establish autophagy as a major cellular pathway required for uterine receptivity and decidualization in both mice and human ESCs.

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Progesterone-Dependent Induction of Phospholipase C-Related Catalytically Inactive Protein 1 (PRIP-1) in Decidualizing Human Endometrial Stromal Cells

Endocrinology ◽

10.1210/en.2015-1914 ◽

2016 ◽

Vol 157 (7) ◽

pp. 2883-2893 ◽

Cited By ~ 16

Author(s):

Joanne Muter ◽

Paul J. Brighton ◽

Emma S. Lucas ◽

Lauren Lacey ◽

Anatoly Shmygol ◽

...

Keyword(s):

Phospholipase C ◽

Stromal Cells ◽

Nuclear Translocation ◽

Scaffold Protein ◽

Trophoblast Invasion ◽

Differentiation Markers ◽

Endometrial Stromal Cells ◽

Decidual Cells ◽

Phosphoinositide 3 Kinase ◽

Inactive Protein

Decidualization denotes the transformation of endometrial stromal cells into specialized decidual cells. In pregnancy, decidual cells form a protective matrix around the implanting embryo, enabling coordinated trophoblast invasion and formation of a functional placenta. Continuous progesterone (P4) signaling renders decidual cells resistant to various environmental stressors, whereas withdrawal inevitably triggers tissue breakdown and menstruation or miscarriage. Here, we show that PLCL1, coding phospholipase C (PLC)-related catalytically inactive protein 1 (PRIP-1), is highly induced in response to P4 signaling in decidualizing human endometrial stromal cells (HESCs). Knockdown experiments in undifferentiated HESCs revealed that PRIP-1 maintains basal phosphoinositide 3-kinase/Protein kinase B activity, which in turn prevents illicit nuclear translocation of the transcription factor forkhead box protein O1 and induction of the apoptotic activator BIM. By contrast, loss of this scaffold protein did not compromise survival of decidual cells. PRIP-1 knockdown did also not interfere with the responsiveness of HESCs to deciduogenic cues, although the overall expression of differentiation markers, such as PRL, IGFBP1, and WNT4, was blunted. Finally, we show that PRIP-1 in decidual cells uncouples PLC activation from intracellular Ca2+ release by attenuating inositol 1,4,5-trisphosphate signaling. In summary, PRIP-1 is a multifaceted P4-inducible scaffold protein that gates the activity of major signal transduction pathways in the endometrium. It prevents apoptosis of proliferating stromal cells and contributes to the relative autonomy of decidual cells by silencing PLC signaling downstream of Gq protein-coupled receptors.

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The Role of Decidual Subpopulations in Implantation, Menstruation and Miscarriage

Frontiers in Reproductive Health ◽

10.3389/frph.2021.804921 ◽

2021 ◽

Vol 3 ◽

Author(s):

Joanne Muter ◽

Chow-Seng Kong ◽

Jan J. Brosens

Keyword(s):

Stromal Cells ◽

Acute Stress ◽

Embryo Implantation ◽

Recurrence Risk ◽

Tissue Remodelling ◽

Endometrial Stromal Cells ◽

Decidual Cells ◽

Unk Cells ◽

Acute Stress Response

In each menstrual cycle, the endometrium becomes receptive to embryo implantation while preparing for tissue breakdown and repair. Both pregnancy and menstruation are dependent on spontaneous decidualization of endometrial stromal cells, a progesterone-dependent process that follows rapid, oestrogen-dependent proliferation. During the implantation window, stromal cells mount an acute stress response, which leads to the emergence of functionally distinct decidual subsets, reflecting the level of replication stress incurred during the preceding proliferative phase. Progesterone-dependent, anti-inflammatory decidual cells (DeC) form a robust matrix that accommodates the conceptus whereas pro-inflammatory, progesterone-resistant stressed and senescent decidual cells (senDeC) control tissue remodelling and breakdown. To execute these functions, each decidual subset engages innate immune cells: DeC partner with uterine natural killer (uNK) cells to eliminate senDeC, while senDeC co-opt neutrophils and macrophages to assist with tissue breakdown and repair. Thus, successful transformation of cycling endometrium into the decidua of pregnancy not only requires continuous progesterone signalling but dominance of DeC over senDeC, aided by recruitment and differentiation of circulating NK cells and bone marrow-derived decidual progenitors. We discuss how the frequency of cycles resulting in imbalanced decidual subpopulations may determine the recurrence risk of miscarriage and highlight emerging therapeutic strategies.

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Exometabolomic analysis of decidualizing human endometrial stromal and perivascular cells

10.1101/2020.08.06.221119 ◽

2020 ◽

Author(s):

Sarah Harden ◽

Jieliang Zhou ◽

Maria Diniz-da-Costa ◽

Emma S. Lucas ◽

Liang Cui ◽

...

Keyword(s):

Time Course ◽

Embryo Implantation ◽

Cellular Reprogramming ◽

Trophoblast Invasion ◽

Perivascular Cells ◽

Endometrial Stromal Cells ◽

Signalling Molecules ◽

Spatiotemporal Information ◽

Decidual Cells ◽

Acid Metabolites

ABSTRACTDifferentiation of endometrial fibroblasts into specialized decidual cells controls embryo implantation and transforms the cycling endometrium into a semi-permanent, immune-protective matrix that accommodates the placenta throughout pregnancy. This process starts during the midluteal phase of the menstrual cycle with decidual transformation of perivascular cells (PVC) surrounding the terminal spiral arterioles and endometrial stromal cells (EnSC) underlying the luminal epithelium. Decidualization involves extensive cellular reprogramming and acquisition of a secretory phenotype, essential for coordinated placental trophoblast invasion. Secreted metabolites are an emerging class of signalling molecules. Here, we used liquid chromatography-mass spectrometry to characterise the dynamic changes in metabolite secretion (exometabolome) of primary PVC and EnSC decidualized over 8 days. We identified 79 annotated metabolites differentially secreted upon decidualization, including prostaglandin, sphingolipid, and hyaluronic acid metabolites. Secreted metabolites encompassed 21 metabolic pathways, most prominently glycerolipid and pyrimidine metabolism. Although temporal exometabolome changes were comparable between decidualizing PVC and EnSC, 32 metabolites were differentially secreted across the decidualization time-course. Further, targeted metabolomics demonstrated a conspicuous difference in xanthine secretion between decidualized PVC and EnSC. Taken together, our findings indicate that the metabolic footprints generated by different decidual subpopulations encode spatiotemporal information that may be important for optimal embryo implantation.

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Impaired decidualization caused by downregulation of circadian clock gene BMAL1 contributes to human recurrent miscarriage†

Biology of Reproduction ◽

10.1093/biolre/ioz063 ◽

2019 ◽

Vol 101 (1) ◽

pp. 138-147 ◽

Cited By ~ 8

Author(s):

Shijian Lv ◽

Na Wang ◽

Jin Ma ◽

Wei-Ping Li ◽

Zi-Jiang Chen ◽

...

Keyword(s):

Stromal Cells ◽

Recurrent Miscarriage ◽

First Trimester ◽

Subsequent Pregnancy ◽

Tissue Inhibitors Of Metalloproteinases ◽

Trophoblast Invasion ◽

Intrauterine Pregnancy ◽

Endometrial Stromal Cells ◽

Aryl Hydrocarbon

Abstract Recurrent miscarriage (RM) is characterized by two or more consecutive losses of a clinically established intrauterine pregnancy at early gestation. To date, the etiology of RM remains poorly understood. Impaired decidualization is thought to predispose women to subsequent pregnancy failure. The transcriptional factor brain and muscle aryl hydrocarbon receptor nuclear translocator-like (BMAL1) controls circadian rhythms and regulates a very large diversity of physiological processes. BMAL1 is essential for fertility. Here, we investigated the expression and function of BMAL1 in human decidualization and its relation with RM. A total of 39 decidua samples were collected. We also examined human endometrial stromal cells (HESCs) and primary endometrial stromal cells (ESCs), and primary decidual stromal cells (DSCs) isolated from decidua of first-trimester pregnancies. Compared to normal pregnant women, the expression of BMAL1 was reduced in the decidual tissues from individuals with RM. After in vitro induction of decidualization, the transcription of BMAL1 in both HESCs and primary ESCs was increased. This is in line with the relatively higher expression of BMAL1 in DSCs than in ESCs. Silencing of BMAL1 resulted in impaired decidualization. Moreover, levels of tissue inhibitors of metalloproteinases (TIMPs) increased significantly upon decidualization. Further experiments demonstrated that BMAL1 silencing curtails the ability of DSCs to restrict excessive trophoblast invasion via downregulation of TIMP3. Our study demonstrates a functional role for BMAL1 during decidualization: the downregulation of BMAL1 in RM leads to impaired decidualization and aberrant trophoblast invasion by regulating TIMP3 and consequently predisposing individuals for RM.

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Decorin production by the human decidua: role in decidual cell maturation

MHR Basic science of reproductive medicine ◽

10.1093/molehr/gaaa058 ◽

2020 ◽

Vol 26 (10) ◽

pp. 784-796

Author(s):

C D Halari ◽

P Nandi ◽

M J Jeyarajah ◽

S J Renaud ◽

P K Lala

Keyword(s):

Growth Factor ◽

Stromal Cells ◽

Binding Protein ◽

Cell Maturation ◽

Decidual Cell ◽

Insulin Like Growth Factor ◽

Endometrial Stromal Cells ◽

Human Trophoblast ◽

Factor Binding ◽

Decidual Cells

Abstract Decidualization involves the proliferation and differentiation of fibroblast-like endometrial stromal cells into epithelioid-shaped and secretory ‘decidual’ cells in response to steroid hormones. Human decidual cells produce insulin-like growth factor-binding protein-1 and prolactin (PRL), two well-recognized markers of decidual cell maturation and a proteoglycan decorin (DCN). We reported that DCN restrains the human trophoblast renewal, migration, invasion and endovascular differentiation needed for uterine arterial remodeling during normal pregnancy. DCN overproduction by the decidua is associated with a hypo-invasive placenta and a serious pregnancy disorder, pre-eclampsia (PE). Furthermore, elevated maternal plasma DCN levels during the second trimester is a predictive biomarker of PE. While these paracrine roles of decidua-derived DCN on trophoblast physiology and pathology have been well-defined, it remains unknown whether DCN plays any autocrine role in decidual cell development. The objectives of this study were to examine: the kinetics of DCN production during decidualization of human endometrial stromal cells; gestational age-related changes in DCN production by the first trimester decidua; and a possible autocrine role of DCN on decidual cell maturation. We found that DCN production is enhanced during decidualization of both primary and immortalized human endometrial stromal cells in vitro and during early gestation in decidual samples tested ex vivo, and that it is important for endometrial stromal cell maturation into a decidual phenotype. Decorin-depleted human endometrial stromal cells exposed to decidualizing stimuli failed to mature fully, as evidenced by fibroblastoid morphology, reduced insulin-like growth factor-binding protein-1 and PRL expression, and reduction in cellular ploidy. We identified heart and neural crest derivatives-expressed protein 2, and progesterone receptor as potential downstream mediators of DCN effects.

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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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Exometabolomic Analysis of Decidualizing Human Endometrial Stromal and Perivascular Cells

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.626619 ◽

2021 ◽

Vol 9 ◽

Author(s):

Sarah L. Harden ◽

Jieliang Zhou ◽

Seley Gharanei ◽

Maria Diniz-da-Costa ◽

Emma S. Lucas ◽

...

Keyword(s):

Time Course ◽

Embryo Implantation ◽

Cellular Reprogramming ◽

Trophoblast Invasion ◽

Perivascular Cells ◽

Endometrial Stromal Cells ◽

Time Resolved ◽

Spatiotemporal Information ◽

Decidual Cells ◽

Purine Pathway

Differentiation of endometrial fibroblasts into specialized decidual cells controls embryo implantation and transforms the cycling endometrium into a semi-permanent, immune-protective matrix that accommodates the placenta throughout pregnancy. This process starts during the midluteal phase of the menstrual cycle with decidual transformation of perivascular cells (PVC) surrounding the terminal spiral arterioles and endometrial stromal cells (EnSC) underlying the luminal epithelium. Decidualization involves extensive cellular reprogramming and acquisition of a secretory phenotype, essential for coordinated placental trophoblast invasion. Secreted metabolites are an emerging class of signaling molecules, collectively known as the exometabolome. Here, we used liquid chromatography-mass spectrometry to characterize and analyze time-resolved changes in metabolite secretion (exometabolome) of primary PVC and EnSC decidualized over 8 days. PVC were isolated using positive selection of the cell surface marker SUSD2. We identified 79 annotated metabolites differentially secreted upon decidualization, including prostaglandin, sphingolipid, and hyaluronic acid metabolites. Secreted metabolites encompassed 21 metabolic pathways, most prominently glycerolipid and pyrimidine metabolism. Although temporal exometabolome changes were comparable between decidualizing PVC and EnSC, 32 metabolites were differentially secreted across the decidualization time-course. Further, targeted metabolomics demonstrated significant differences in secretion of purine pathway metabolites between decidualized PVC and EnSC. Taken together, our findings indicate that the metabolic footprints generated by different decidual subpopulations encode spatiotemporal information that may be important for optimal embryo implantation.

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The actions of resveratrol in decidualizing endometrium: acceleration or inhibition?†

Biology of Reproduction ◽

10.1093/biolre/ioaa172 ◽

2020 ◽

Vol 103 (6) ◽

pp. 1152-1156

Author(s):

Keiji Kuroda ◽

Asako Ochiai ◽

Jan J Brosens

Keyword(s):

Stromal Cells ◽

Ovarian Function ◽

Embryo Implantation ◽

Clinical Effects ◽

Polyphenolic Compound ◽

Endometrial Stromal Cells ◽

Decidual Cells ◽

Multistep Process ◽

Drug Interventions

Abstract Resveratrol, a natural polyphenolic compound, is widely studied for its anti-inflammatory and antisenescent properties. Recently, two studies reported seemingly conflicting findings on the actions of resveratrol on decidualization of human endometrial stromal cells (HESCs). One study by Ochiai et al. demonstrated that resveratrol inhibits decidual transformation of primary cultured HESCs. The other study by Mestre Citrinovitz et al., showed that resveratrol enhances decidualization of HESCs in culture. At a glance, the reason for these opposing observations seems puzzling. However, recent studies demonstrated that decidualization is a multistep process, which starts with an acute proinflammatory stress response that lasts for several days and is followed by the emergence of stress-resistant decidual cells as well as senescent decidual cells. The balance between these decidual subpopulations may determine if the cycling endometrium can successfully transition into the decidua of pregnancy upon embryo implantation. Here, we explore the importance of timing of drugs aimed at modulating the decidual response. We posit that resveratrol treatment during the initial proinflammatory decidual phase, i.e., coinciding with the implantation window in vivo, inhibits decidual transformation of the endometrium. However, when given after the initial phase, resveratrol may promote decidualization by inhibiting decidual senescence. Further, if restricted to the proliferative phase, resveratrol may promote ovarian function without adversely impacting on embryo implantation or decidualization. Thus, failure to align drug interventions with the correct phase of the menstrual cycle may negate beneficial clinical effects and results in adverse reproductive outcomes.

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