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To study the cyclic fertilin peptide effects on preimplantation human embryogenesis. Cyclic fertilin peptide reproduces the structure of the binding site of the sperm Fertilin β (also named A Disintegrin and Metalloprotease 2: ADAM2) disintegrin domain. It binds to the oocyte membrane and increases sperm-oocyte fusion index in human and fertilization rate in mouse, providing healthy pups. It also improves human oocyte maturation and chromosome segregation in meiosis I and binds to human embryo blastomeres, suggesting that it has a membrane receptor.Thawed human embryos at the 3 to 4 cells stage were randomly included in a dose-response study with cyclic fertilin peptide. Inner cell mass (ICM), trophectoderm (TE), and total cell numbers were evaluated in top- and good-quality blastocysts.The study was performed in an academic hospital and research laboratory.Human embryos donated for research. This project was approved by the French "Agence de la Biomédecine."Immunofluorescence and tissue-specific gene expression analysis, using Clariom D microarrays, were performed to study its mechanism of action.Cyclic fertilin peptide improves blastocyst formation by almost 20%, the concentration of 1 μM being the lowest most efficient concentration. It significantly increases twice the TE cell number, without modifying the ICM. It increases the in vitro hatching rate from 14% to 45%.Cyclic fertilin peptide stimulates TE growth. In the ICM, it induces transcriptional activation of intracellular protein and vesicle-mediated transport.Cyclic fertilin peptide dramatically improves human embryo development potential. It could be used to supplement culture medium and improve the in vitro human embryo development. Starting supplementation immediately after fertilization, instead of day 2, could significantly upgrade assisted reproductive technology outcome.

Simple Summary Endometrial cancer (EC) is the 2nd most common gynecologic cancer worldwide. MicroRNAs (miRNAs) are small noncoding RNAs that contribute to epigenetic regulation. The objective of this systematic review is to summarize our current knowledge on the role of miRNAs in the epigenetic deregulation of tumor-related genes in EC. It includes all miRNAs reported to be involved in EC including their roles in DNA methylation and RNA-associated silencing. This systematic review should be useful for development of novel strategies to improve diagnosis and risk assessment as well as for new treatments aimed at miRNAs, their target genes or DNA methylation. Abstract The objective of this systematic review is to summarize our current knowledge on the influence of miRNAs in the epigenetic deregulation of tumor-related genes in endometrial cancer (EC). We conducted a literature search on the role of miRNAs in the epigenetic regulation of EC applying the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The following terms were used: microRNA, miRNA, miR, endometrial cancer, endometrium, epigenetic, epimutation, hypermethylation, lynch, deacetylase, DICER, novel biomarker, histone, chromatin. The miRNAs were classified and are presented according to their function (tumor suppressor or onco-miRNA), their targets (when known), their expression levels in EC tissue vs the normal surrounding tissue, and the degree of DNA methylation in miRNA loci and CpG sites. Data were collected from 201 articles, including 190 original articles, published between November 1, 2008 and September 30, 2020 identifying 313 different miRNAs implicated in epigenetic regulation of EC. Overall, we identified a total of 148 miRNAs with decreased expression in EC, 140 miRNAs with increased expression in EC, and 22 miRNAs with discordant expression levels. The literature implicated different epigenetic phenomena including altered miRNA expression levels (miR-182, -230), changes in the methylation of miRNA loci (miR-34b, -129-2, -130a/b, -152, -200b, -625) and increased/decreased methylation of target genes (miR-30d,-191). This work provides an overview of all miRNAs reported to be involved in epigenetic regulation in EC including DNA methylation and RNA-associated silencing. These findings may contribute to novel strategies in diagnosis, risk assessment, and treatments aimed at miRNAs, their target genes or DNA methylation.

Group A Streptococcus (GAS), a Gram-positive human-specific pathogen, yields 517,000 deaths annually worldwide, including 163,000 due to invasive infections and among them puerperal fever. Before efficient prophylactic measures were introduced, the mortality rate for mothers during childbirth was approximately 10%; puerperal fever still accounts for over 75,000 maternal deaths annually. Yet, little is known regarding the factors and mechanisms of GAS invasion and establishment in postpartum infection. We characterized the early steps of infection in an ex vivo infection model of the human decidua, the puerperal fever portal of entry. Coordinate analysis of GAS behavior and the immune response led us to demonstrate that (a) GAS growth was stimulated by tissue products; (b) GAS invaded tissue and killed approximately 50% of host cells within 2 hours, and these processes required SpeB protease and streptolysin O (SLO) activities, respectively; and (c) GAS impaired the tissue immune response. Immune impairment occurred both at the RNA level, with only partial induction of the innate immune response, and protein level, in an SLO- and SpeB-dependent manner. Our study indicates that efficient GAS invasion of the decidua and the restricted host immune response favored its propensity to develop rapid invasive infections in a gynecological-obstetrical context.

Two major obstetric diseases, preeclampsia (PE), a pregnancy-induced endothelial dysfunction leading to hypertension and proteinuria, and intra-uterine growth-restriction (IUGR), a failure of the fetus to acquire its normal growth, are generally triggered by placental dysfunction. Many studies have evaluated gene expression deregulations in these diseases, but none has tackled systematically the role of alternative splicing. In the present study, we show that alternative splicing is an essential feature of placental diseases, affecting 1060 and 1409 genes in PE vs controls and IUGR vs controls, respectively, many of those involved in placental function. While in IUGR placentas, alternative splicing affects genes specifically related to pregnancy, in preeclamptic placentas, it impacts a mix of genes related to pregnancy and brain diseases. Also, alternative splicing variations can be detected at the individual level as sharp splicing differences between different placentas. We correlate these variations with genetic variants to define splicing Quantitative Trait Loci (sQTL) in the subset of the 48 genes the most strongly alternatively spliced in placental diseases. We show that alternative splicing is at least partly piloted by genetic variants located either in cis (52 QTL identified) or in trans (52 QTL identified). In particular, we found four chromosomal regions that impact the splicing of genes in the placenta. The present work provides a new vision of placental gene expression regulation that warrants further studies. Supplementary Information The online version contains supplementary material available at 10.1007/s00439-020-02248-x.

Preeclampsia is a major, frequent and potentially severe condition of pregnancy, characterized by severe hypertension and proteinuria. In this review, we describe recent advances in understanding the pathology, and discuss the long-term impacts on maternal vascular health. Next, we describe the genetic, epigenetic and immunological basis of preeclampsia. We describe the links between preeclampsia and oxidative stress in placental (trophoblast) and endothelial cells. We mention cellular and animal models commonly used to decipher modified pathophysiological pathways in a preeclamptic pregnancy compared to a normal pregnancy. Finally, we discuss the therapeutic options, readily available or in development, to improve the monitoring of pregnancies, the health of patients and that of children born from preeclamptic pregnancies.

Summary STOX1 is a transcription factor involved in preeclampsia and Alzheimer disease. We show that the knock-down of the gene induces rather mild effect on gene expression in trophoblast cell lines (BeWo). We identified binding sites of STOX1 shared by the two major isoforms, STOX1A and STOX1B. Profiling gene expression of cells overexpressing either STOX1A or STOX1B, we identified genes downregulated by both isoforms, with a STOX1 binding site in their promoters. Among those, STOX1-induced Annexin A1 downregulation led to abolished membrane repair in BeWo cells. By contrast, overexpression of STOX1A or B has opposite effects on trophoblast fusion (acceleration and inhibition, respectively) accompanied by syncytin genes deregulation. Also, STOX1A overexpression led to abnormal regulation of oxidative and nitrosative stress. In sum, our work shows that STOX1 isoform imbalance is a cause of gene expression deregulation in the trophoblast, possibly leading to placental dysfunction and preeclampsia., Graphical Abstract, Highlights • STOX1, involved in preeclampsia, recognizes specific DNA sequences • Downregulation of STOX1 isoforms has limited effect on trophoblast gene expression • Overexpression of STOX1A and B leads to downregulation of some genes • Overexpression of STOX1A or STOX1B leads to opposite effects on other genes, Reproductive Medicine; Molecular Biology; Developmental Biology

Group A Streptococcus (GAS), a Gram-positive human-specific pathogen yields 517,000 deaths annually worldwide, including 163,000 due to invasive infections and among them puerperal fever. GAS is their most feared etiologic agent. Puerperal fever still accounts for more than 75,000 maternal deaths annually and before the introduction of efficient prophylactic measures 10% childbirths were followed by the mother’s death. Yet little is known regarding GAS invasive infection establishment or GAS efficiency in causing postpartum infection. To characterize its early steps, we set up coordinated analyses of ex vivo infection of the human decidua, the puerperal fever portal of entry. We analyzed GAS behavior and the immune response triggered. We demonstrate that GAS (i) benefits from tissue secreted products to multiply; (ii) invades the tissue and leads to the death of half the cells within two hours via SpeB protease and Streptolysin O activities, respectively; (iii) impairs the tissue immune response. Immune impairment occurs both at the RNA level, with the induction of only a restricted immediate innate immune response, and at the protein level, in a SLO- and SpeB-dependent manner. Our study indicates that GAS efficient decidua invasion and immune response restraint favor its propensity to develop rapid invasive infections in a gynecological-obstetrical context.

Adverse long-term cardiovascular (CV) consequences of PE are well established in women. However, the mechanism responsible for that risk remains unknown. Here, we mated wild-type female mice of the FVB/N strain to STOX1A-overexpressing mice to mimic severe PE and investigated the long-term consequences on the maternal cardiovascular system. Ultrasonography parameters were analyzed in mice before pregnancy and at 3 and 6 months post-pregnancy. At 6 months post-pregnancy, cardiac stress test induced by dobutamine injection revealed an abnormal ultrasonography Doppler profile in mice with previous PE. Eight months post-pregnancy, the heart, endothelial cells (ECs) and plasma of females were analyzed and compared to controls. The heart of mice with PE showed left-ventricular hypertrophy associated with altered histology (fibrosis). Transcriptomic analysis revealed the deregulation of 1149 genes in purified ECs and of 165 genes in the hearts, many being involved in heart hypertrophy. In ECs, the upregulated genes were associated with inflammation and cellular stress. Systems biology analysis identified interleukin 6 (IL-6) as a hub gene connecting these pathways. Plasma profiling of 33 cytokines showed that, 8 of them (Cxcl13, Cxcl16, Cxcl11, IL-16, IL-10, IL-2, IL-4 and Ccl1) allowed to discriminate mice with previous PE from controls. Thus, PE triggers female long-term CV consequences on the STOX1 mouse model.

In this review, we comprehensively present the function of epigenetic regulations in normal placental development as well as in a prominent disease of placental origin, preeclampsia (PE). We describe current progress concerning the impact of DNA methylation, non-coding RNA (with a special emphasis on long non-coding RNA (lncRNA) and microRNA (miRNA)) and more marginally histone post-translational modifications, in the processes leading to normal and abnormal placental function. We also explore the potential use of epigenetic marks circulating in the maternal blood flow as putative biomarkers able to prognosticate the onset of PE, as well as classifying it according to its severity. The correlation between epigenetic marks and impacts on gene expression is systematically evaluated for the different epigenetic marks analyzed.

Introduction: Endometrial cancer (EC) is the most important gynecological cancer in terms of incidence. microRNAs (miRs), which are post-transcriptional regulators implicated in a variety of cellular functions including carcinogenesis, are particularly attractive candidates as biomarkers. Indeed, several studies have shown that the miR expression pattern appears to be associated with prognostic factors in EC. Our objective is to review the current knowledge of the role of miRs in carcinogenesis and tumor progression and their association with the prognosis of endometrial cancer. Materials and Method: We performed a literature search for miR expression in EC using MEDLINE, PubMed (the Internet portal of the National Library of Medicine) and The Cochrane Library, Cochrane databases “Cochrane Reviews” and “Clinical Trials” using the following keywords: microRNA, endometrial cancer, prognosis, diagnosis, lymph node, survival, plasma, FFPE (formalin-fixed, paraffin-embedded). The miRs were classified and presented according to their expression levels in cancer tissue in relation to different prognostic factors. Results: Data were collected from 74 original articles and 8 literature reviews which described the expression levels of 261 miRs in ECs, including 133 onco-miRs, 110 miR onco-suppressors, and 18 miRs with discordant functions. The review identified 30 articles studying the expression pattern of miR in neoplastic endometrial tissue compared to benign and/or hyperplastic tissues, 12 articles detailing the expression profile of miRs as a function of lymph node status, and 14 articles that detailed the expression pattern of miRs in endometrial tumor tissue according to overall survival or in the absence of recurrence. Conclusions: The findings presented here suggest that miR analysis merits a role as a prognostic factor in the management of patients with endometrial cancer.

STOX1 is a transcription factor involved in two complex and frequent human diseases, preeclampsia and Alzheimer disease. However, precisely how STOX1 controls gene expression is still not well known. In this study, we identified binding sites of STOX1shared by the two major isoforms of the protein, STOX1A and STOX1B. Using newly generated villous trophoblast cell lines overexpressing either STOX1A or STOX1B, we identified a group of genes down-regulated by both isoforms, which contains one of the STOX1 binding sites that we identified in their promoters. Amongst those gene Annexin A1 (a major protein for membrane repair) was drastically down-regulated. We show that membrane repair was consistently abolished in BeWo cells overexpressing STOX1 isoforms. We then studied the effect of STOX on trophoblast fusion, a major event in villous trophoblast biology. STOX1A and B overexpression had opposite effects on trophoblast fusion: STOX1A accelerated the process and STOX1B prevented it in particular by deregulating syncytin genes which are endoretroviruses crucial for trophoblast fusion in mammals and other viviparous vertebrates. We also show that STOX1A overexpression led to abnormal regulation of oxidative and nitrosative stress in the cells. In sum STOX1 isoform imbalance appears as a possible cause of deregulation of gene expression in trophoblast, possibly leading to placental dysfunction and preeclampsia.

Group A Streptococcus (GAS) is a human-specific pathogen responsible for a wide range of diseases, ranging from superficial to life-threatening invasive infections, including endometritis, and autoimmune sequelae. GAS strains express a vast repertoire of virulence factors that varies depending on the strain genotype, and many adhesin proteins that enable GAS to adhere to host cells are restricted to some genotypes. GAS emm28 is the third most prevalent genotype in invasive infections in France and is associated with gyneco-obstetrical infections. emm28 strains harbor R28, a cell wall–anchored surface protein that has previously been reported to promote adhesion to cervical epithelial cells. Here, using cellular and biochemical approaches, we sought to determine whether R28 supports adhesion also to other cells and to characterize its cognate receptor. We show that through its N-terminal domain, R28(Nt), R28 promotes bacterial adhesion to both endometrial–epithelial and endometrial–stromal cells. R28(Nt) was further subdivided into two domains, and we found that both are involved in cell binding. R28(Nt) and both subdomains interacted directly with the laminin-binding α3β1, α6β1, and α6β4 integrins; interestingly, these bindings events did not require divalent cations. R28 is the first GAS adhesin reported to bind directly to integrins that are expressed in most epithelial cells. Finally, R28(Nt) also promoted binding to keratinocytes and pulmonary epithelial cells, suggesting that it may be involved in supporting the prevalence in invasive infections of the emm28 genotype.

Preeclampsia is a major, frequent and potentially severe condition of pregnancy, characterized by severe hypertension and proteinuria. In this review, we describe recent advances in understanding the pathology, and discuss the long-term impacts on maternal vascular health. Next, we describe the genetic, epigenetic and immunological basis of preeclampsia. We describe the links between preeclampsia and oxidative stress in placental (trophoblast) and endothelial cells. We mention cellular and animal models commonly used to decipher modified pathophysiological pathways in a preeclamptic pregnancy compared to a normal pregnancy. Finally, we discuss the therapeutic options, readily available or in development, to improve the monitoring of pregnancies, the health of patients and that of children born from preeclamptic pregnancies.

Previously, we reported endometriotic-like decidual lesions in contact with the fetal membranes (FMs) in 11 pregnant women with severe endometriosis. In this report, an extensive histomorphological analysis was performed on the FMs of 19 pregnant women with deep infiltrating endometriosis (DIE) at term pregnancy and who delivered by cesarean delivery before labor. On gross examination, all samples showed increased thickness, de novo microvessel formation, and small-size excrescences distributed along the membrane circumference. Histological examination of FM fragments sampled from the placenta edges or from the cesarean incision line showed fibrinoid necrosis and connective tissue accumulation in the amnion, chorion, and decidual layers in most of the 19 women with DIE. Papillary tufting and epithelial cell multilayering at the surface of the amnion layer were found in 3 of the 19 women with DIE. In 14 of the 19 women with DIE, the trophoblastic layer was disrupted by dense extracellular material, degenerative villi, and inflammatory infiltrates. Cystic gland-like structures were found in the decidual layer in all the 19 women with DIE, which were surrounded by irradiating small vessels and scattered inflammatory cells. The relationship between these peculiar histological changes and the endometriotic status of the pregnant women is still unclear. Sustained examination of FMs in women with DIE is needed to fully evaluate the defaults in these tissue structures and to establish whether these defaults have clinical impact on the pregnancy course.

Study question Are the fetal membranes of women affected with endometriosis similar to those from disease-free women? Summary answer Decidua of women with endometriosis is able to generate endometriotic-like lesions in contact with the fetal membranes. What is known already Eutopic endometrium of women affected with endometriosis presents compromised properties. Endometrium undergoes decidualisation to accept and to further control the conceptus development during pregnancy. Decidualized endometrium is in close contact with the chorionic membrane and forms the choriodecidual layer, a major maternal-fetal interface. Study design, size, duration This is a laboratory case-control study involving diseased versus control samples. Eleven case samples and 11 control samples were collected from women in a tertiary care/research center between November 2011 and December 2013. Participants/materials, setting, methods Participants were consecutive pregnant women affected with confirmed endometriosis and disease free women, who underwent Cesarean section before labor for obstetrical indication. The choriodecidual tissues were characterized using histology, immunohistochemistry, transcriptomic and whole genome CpG methylation analyses. Main results and the role of chance We demonstrate for the first time the presence of endometriotic-like lesions within the decidual side of the choriodecidua of the fetal membranes from women affected with severe endometriosis. Fetal membranes from women affected with endometriosis exhibited glandular components in the choriodecidual layer surrounded by enlarged decidualized cells disseminated along the entire membrane surface. Significant deregulation (variation of expression ≥2, P-value ≤0.05) was observed for 2773 genes known to be enriched in processes involved in glandular function, endocrine and nervous system, neoangiogenesis, and autoimmune disease. CpG methylation analysis revealed 5999 differentially methylated regions with a P-value ≤0.05. Limitations, reasons for caution We studied women who delivered at term by Cesarean section before labor, following an uneventful pregnancy. Notwithstanding this, one cannot exclude that the presence of disseminated endometriotic lesions within the choriodecidual layer of the fetal membranes may disturb the anatomical integrity and/or the function of the membranes in some women with endometriosis. Wider implications of the findings Our results shed new light on the capability of the diseased decidua to develop lesions not only at ectopic autologous locations, but also on the semi-allogenous fetal membranes, a particularly immunotolerant environment.

Aims: Storkhead box 1 (STOX1) is a winged-helix transcription factor that is implicated in the genetic forms of a high-prevalence human gestational disease, pre-eclampsia. STOX1 overexpression confers pre-eclampsia-like transcriptomic features to trophoblastic cell lines and pre-eclampsia symptoms to pregnant mice. The aim of this work was to evaluate the impact of STOX1 on free radical equilibrium and mitochondrial function, both in vitro and in vivo. Results: Transcriptome analysis of STOX1-transgenic versus nontransgenic placentas at 16.5 days of gestation revealed alterations of mitochondria-related pathways. Placentas overexpressing STOX1 displayed altered mitochondrial mass and were severely biased toward protein nitration, indicating nitroso-redox imbalance in vivo. Trophoblast cells overexpressing STOX1 displayed an increased mitochondrial activity at 20% O2 and in hypoxia, despite reduction of the mitochondrial mass in the former. STOX1 overexpression is, therefore, associated with hyperactive mitochondria, resulting in increased free radical production. Moreover, nitric oxide (NO) production pathways were activated, resulting in peroxynitrite formation. At low oxygen pressure, STOX1 overexpression switched the free radical balance from reactive oxygen species (ROS) to reactive nitrogen species (RNS) in the placenta as well as in a trophoblast cell line. Innovation: In pre-eclamptic placentas, NO interacts with ROS and generates peroxynitrite and nitrated proteins as end products. This process will deprive the maternal organism of NO, a crucial vasodilator molecule. Conclusion: Our data posit STOX1 as a genetic switch in the ROS/RNS balance and suggest an explanation for elevated blood pressure in pre-eclampsia. Antioxid. Redox Signal. 21, 819–834.

Introduction Preeclampsia is a major disease of pregnancy, affecting 2–8% of pregnant women. It is characterized by arterial hypertension and proteinuria occurring from the 20th week of amenorrhea. The STOX1 overexpressing mice develop a preeclamptic syndrome. This model is very relevant to identify potential new actors in preeclampsia because the phenotype is both precocious and severe. Objectives Using this mouse model of severe PE, we aim to identify plasma proteins quantitatively modified during pregnancy with a global proteomics approach in the blood and to validate these proteins on human placental cell models. Methods We provoked preeclamptic gestations in mice by crossing WT females with transgenic STOX1 males. Blood was taken at E6.5 of gestation and plasma was analyzed using ITRAQ mass spectrometry to identify proteins of differential abundance. In parallel, we developed BeWo cells stably overexpressing STOX1. BeWo is a choriocarcinoma cell line in which fusion is induced by forskolin treatment and reproduces therefore the physiology of villous trophoblast that syncytialize into syncytiotrophoblast. We are validating the link between STOX1 overexpression and sLIFR production by qRT-PCR and ELISA in this cell model. Results We identified 15 proteins differentially present, two of which have human homologs: sLIFR (soluble Leukemia Inhibitory Factor Receptor) and TTR (transthyretin) respectively increased and decreased in the preeclamptic mice. In the BeWo cells, after fusion or in the presence of STOX1, alternative splicing of LIFR mRNA increases, leading to more secretion of sLIFR in the supernatant. Discussion An early excess of sLIFR could trap Leukemia Inhibitory Factor (LIF) at early stages of embryo implantation and development. Given the primordial role of LIF in implantation in mice and humans, this could lead to a defective implantation and/or placentation possibly causing embryo resorptions or a preeclamptic phenotype.

Preeclampsia (PE) is a common human-specific pregnancy disorder defined by hypertension and proteinuria during gestation and responsible for maternal and fetal morbimortality. STOX1 , encoding a transcription factor, was the first gene associated with PE as identified by positional cloning approaches. Its overexpression in choriocarcinoma cells mimics the transcriptional consequences of PE in the human placenta. Here, we created transgenic mouse strains overexpressing human STOX1 . Wild-type female mice crossed with transgenic male mice reproduce accurately the symptoms of severe PE: gestational hypertension, proteinuria, and elevated plasma levels of soluble fms-like tyrosine kinase 1 and soluble endoglin. Placental and kidney histology were altered. Symptoms were prevented or alleviated by aspirin treatment. STOX1-overexpressing mice constitute a unique model for studying PE, allow testing therapeutic approaches, and assessing the long-term effects of the preeclamptic syndrome.

Lymphovascular space involvement (LVSI) is a major prognostic factor in type 1 endometrial cancer (EC). However, its use has been criticized because of poor subjectivity. MicroRNA signatures have recently been linked to EC pathologic characteristics. The aim of this study was to evaluate whether microRNA profiles of type 1 EC can be related to LVSI status and used as a tool to adapt therapy.MicroRNA expression was assessed by chip analysis and qRT-PCR in 12 formalin-fixed paraffin-embedded grade 2 EC specimens with positive LVSI and in 12 specimens with negative LVSI. Various statistical analyses, including enrichment analysis and a minimum p-value approach, were performed.The expression levels of microRNAs 34c-5p, -23b-5p, and 23c were significantly lower in the EC with positive LVSI compared to those with negative LVSI. Women with a microRNA-34c-5p fold change0.15 were more likely to have positive LVSI status (92.3%) compared with those with a microRNA-34c-5p fold change0.15 (0.0%), p0.001. Furthermore, women with a microRNA-23b-5p fold change0.51 were more likely to have positive LVSI status (90.0%) compared with those with a microRNA-23b-5p fold change0.51 (21.4%), p=0.003.This was the first study to investigate the relative expression of microRNA in type 1 EC according to LVSI status. This microRNA expression profile may provide a basis for further study of the microRNA function in EC, and be used as a diagnostic tool for LVSI status.

Resume Les membranes fœtales, amnios et chorion, tapissent la cavite amniotique et sont essentielles pour son integrite jusqu’au terme normal de la grossesse. Elles ont une structure pluristratifiee dont la composition garantit leur cohesion et leur elasticite. Leur fonction princeps est de retenir le liquide amniotique fluctuant dans une cavite semi-rigide. Leur limite elastique depend de l’organisation de la matrice extracellulaire et principalement du type de collagene qui la compose. Le feuillet compact de l’amnios, responsable de la limite elastique, contient majoritairement du collagene de type I organise en treillis, lui permettant de s’allonger ou de s’epandre. La couche spongieuse sous-jacente, principalement du collagene de type III, est organisee en un maillage, lâche et riche en proteoglycanes hydrates, qui absorbe les chocs et autorise l’amnios a glisser sur le chorion. La sequence d’evenements entrainant la rupture comprend : (i) la distension des membranes avec perte d’elasticite, (ii) la separation de l’amnios et du chorion, (iii) la fracture du chorion, (iv) la distension de l’amnios qui fait hernie, (v) la rupture de l’amnios. Le mecanisme de rupture a longtemps ete considere comme consecutif aux contractions uterines. Cependant l’observation avant le travail d’une zone de morphologie alteree en regard du col, avec des variations biochimiques (modifications des activites metalloproteases et des proteoglycanes, apoptose…) associees a une faiblesse physique focale suggere une programmation de la rupture avant la parturition. Une meilleure comprehension des mecanismes biochimiques de la rupture des membranes pourra apporter un eclairage nouveau sur les moyens d’anticiper et d’intervenir sur les risques de rupture prematuree.

This is an author-produced version of a manuscript accepted for publication in The Journal of Immunology (The JI). The American Association of Immunologists, Inc. (AAI), publisher of The JI, holds the copyright to this manuscript. This version of the manuscript has not yet been copyedited or subjected to editorial proofreading by The JI; hence, it may differ from the final version published in The JI (online and in print). AAI (The JI) is not liable for errors or omissions in this author-produced version of the manuscript or in any version derived from it. The final, citable version of record can be found at www.jimmunol.org.; International audience; Spontaneous preterm delivery is linked to intrauterine inflammation. Fetal membranes are involved in the inflammatory process as an important source of mediators, and the chorion leave produces high levels of the proinflammatory cytokine TNF-alpha when stimulated by LPS. The transcription factor NF-kappaB is the main regulator of this inflammatory process and controls the production of cytokines by the chorion leave. Phosphodiesterase 4 inhibitors are recognized for their anti-inflammatory and myorelaxant effects. The purpose of this study was to investigate whether PDE4 inhibition affects the LPS signaling in human cultured chorionic cells. We showed that these cells express TLR4, the main LPS receptor, and exhibit a predominant PDE4 activity. Upon LPS challenge, PDE4 activity increases concomitantly to the induction of the specific isoform PDE4B2 and chorionic cells secrete TNF-alpha. LPS induces the nuclear translocation of the NF-kappaB p65 subunit and the activation of three different NF-kappaB complexes in chorionic cells. The presence of the PDE4 inhibitor rolipram reduces the TNF-alpha production and the activation of the three NF-kappaB complexes. These data indicate that the PDE4 family interacts with the LPS signaling pathway during the inflammatory response of chorionic cells. PDE4 selective inhibitors may thus represent a new therapeutic approach in the management of inflammation-induced preterm delivery.

Preeclampsia is a disease of pregnancy involving systemic endothelial dysfunction. However, cardiovascular consequences of preeclampsia are difficult to analyze in humans. The objective of the present study is to evaluate the cardiovascular dysfunction induced by preeclampsia by examining the endothelium of mice suffering of severe preeclampsia induced by STOX1 overexpression. Using Next Generation Sequencing on endothelial cells of mice carrying either transgenic or control embryos, we discovered significant alterations of gene networks involved in inflammation, cell cycle and cardiac hypertrophy. In addition, the heart of the preeclamptic mice revealed cardiac hypertrophy associated with histological anomalies. Bioinformatics comparison of the networks of modified genes in the endothelial cells of the preeclamptic mice and HUVECs exposed to plasma from preeclamptic women identified striking similarities. The cardiovascular alterations in the pregnant mice are comparable to those endured by the cardiovascular system of preeclamptic women. The STOX1 mice could help to better understand the endothelial dysfunction in the context of preeclampsia and guide the search for efficient therapies able to protect the maternal endothelium during the disease and its aftermath.

International audience; Conventional methods used for histologic classification and grading of endometrial cancer (EC) are not sufficient to predict lymph node metastases. microRNA signatures have recently been related to EC pathologic characteristics or prognosis. The aim of this study was to evaluate whether microRNA profiles of grade 1–2 endometrioid adenocarcinomas can be related to nodal status and used as a tool to adapt surgical staging in early-stage EC. microRNA expression was assessed in nine formalin-fixed paraffin-embedded (FFPE) EC primary tumors with positive lymph node and in 27 FFPE EC primary tumors with negative lymph node, matched for grade, stage, and lymphovascular space involvement status. A microarray analysis showed that there was more than a twofold significant difference in the expression of 12 microRNAs between the two groups. A quantitative reverse transcriptase–PCR assay was used to confirm these results: the expression levels of five microRNAs (microRNA-34c-5p, -375, -184, -34c-3p, and -34b-5p) were significantly lower in the EC primary tumor with positive lymph node compared with those with negative lymph node. A minimal P-value approach revealed that women with a microRNA-375-fold change 0.30 (n=1; 4.8%), P=0.001. Furthermore, women with a microRNA 184-fold change 0.30 (n=3; 11.5%), P=0.006. This is the first study investigating the relative expression of mature microRNA genes in early-stage grade 1–2 EC primary tumors according to the nodal status. This microRNA expression profile provides a potential basis for further study of the microRNA function in EC and could be used as a diagnostic tool for nodal status.

Intrauterine infections are important etiological factors of preterm labor. They trigger an increase in proinflammatory cytokines, in particular IL-1beta, that induces a cascade of events resulting in the production of potent effectors of myometrial contractility, such as the prostaglandin E(2) (PGE(2)). Within the smooth muscle cells, contractility is under the control of cAMP content, partly regulated by cAMP-phosphodiesterase 4 (PDE4), the predominant family of PDEs expressed in human myometrium. In the present study, using a model of inflammation of human myometrial cells in culture, we demonstrated that exposing the cells to IL-1beta resulted in a significant up-regulation of PDE4 activity through an increase in PDE4B2 mRNA and protein levels. The IL-1beta-induced PDE4 activity occurs after an increase in PGE(2) production and subsequent cAMP augmentation. Pretreatment with indomethacin or NS 398 completely blocked this long-term effect of IL-1beta, revealing a PGE(2)-dependent pathway. Accordingly, our results demonstrated that the PDE4B2 variant can participate in the regulation of the inflammatory reaction that occurs at term or in preterm labor and leads to myometrial contractions. Knowing the myorelaxant effect of PDE4 inhibitors and the implication of the PDE4B2 in the inflammatory process, this isoform may be an appropriate target for discovering antiinflammatory drugs to manage infection-induced preterm deliveries.

The discovery that degradation and inactivation of the second messengers cAMP and cGMP are mediated by a complex enzymatic machinery has changed our perspective on cyclic nucleotide-mediated processes. In the cell, these second messengers are inactivated by no fewer than 11 distinct families of phosphodiesterases (PDEs). Much is known about the structure and function of these enzymes, their complex subcellular distribution and regulation. Yet, their potential as targets for therapeutic intervention in a broad range of endocrine abnormalities still needs to be investigated. This review explores the involvement of PDEs in the regulation of intracellular signaling and focuses on the known and potential roles that are of interest to endocrinologists.

Elevation of cAMP content resulting from stimulation of the receptor-adenylyl cyclase complex is involved in maintaining the quiescence of the human myometrium during pregnancy. The magnitude of this elevation is critically influenced by the rate of cAMP hydrolysis by phosphodiesterase (PDE) isoenzymes. In the present study we report that in term myometrium, enhanced cAMP-specific PDE4 activity takes part in the heterologous desensitization to the beta-mimetic, salbutamol. Indeed, pretreatment with a PDE4-selective inhibitor potentiates the relaxant effect of salbutamol on myometrial strips of women at term. Furthermore, the reduced relaxant effect of salbutamol after long-term treatment of myometrial strips with PGE2, a potent myometrial effector, can be reversed by PDE4 inhibition. Using a model of cultured myometrial cells, we also demonstrated that PGE2 is able to up-regulate PDE4 activity, at least through the induction of synthesis of PDE4B and PDE4D short forms, which, in turn, dampen the cAMP accumulation provoked by the stimulation of adenylyl cyclase. Such data suggest that in late pregnancy endogenous PGE2 might up-regulate PDE4 activity and lessen the responsiveness of myometrium to beta-mimetic activation. Accordingly, coapplication of a selective PDE4 inhibitor might greatly improve the usefulness of beta-mimetic in tocolysis.

Preeclampsia is a pregnancy disorder characterized by hypertension and proteinuria. In preeclampsia, the placenta releases factors into the maternal circulation that cause a systemic endothelial dysfunction. Herein, we investigated the effects of plasma from women with preeclamptic and normal pregnancies on the transcriptome of an immortalized human umbilical vein endothelial cell line. The cells were exposed for 24 hours to preeclamptic or normal pregnancy plasma and their transcriptome was analyzed using Agilent microarrays. A total of 116 genes were found differentially expressed: 71 were up-regulated and 45 were down-regulated. In silico analysis revealed significant consistency and identified four functional categories of genes: mitosis and cell cycle progression, anti-apoptotic, fatty acid biosynthesis, and endoplasmic reticulum stress effectors. Moreover, several genes involved in vasoregulation and endothelial homeostasis showed modified expression, including EDN1, APLN, NOX4, and CBS. Promoter analysis detected, among the up-regulated genes, a significant overrepresentation of genes containing activation protein-1 regulatory sites. This correlated with down-regulation of JDP2, a gene encoding a repressor of activation protein-1. The role of JDP2 in the regulation of a subset of genes in the human umbilical vein endothelial cells was confirmed by siRNA inhibition. We characterized transcriptional changes induced by preeclamptic plasma on human umbilical vein endothelial cells, and identified, for the first time to our knowledge, JDP2 as a regulator of a subset of genes modified by preeclamptic plasma.

STOX1, un gène codant un facteur de transcription, a été lié à la prééclampsie, une pathologierépandue et spécifiquement humaine de la grossesse caractérisée par une hypertension et une protéinurie gestationnelles, et responsable de mortalitéet de morbidité maternelles et fœtales. Cette étudevise à mieux comprendre le rôle de STOX1 in vivo.Pour étudier son implication dans la pathologie,nous avons créé des souris transgéniques surexprimant le STOX1 humain. Des souris femellescontrôles ont été croisées avec des mâles transgéniques afin de limiter l’expression du transgène àl’unité fœto-placentaire et empêcher ainsi un effetdirect du transgène sur l’organisme maternel (reinet endothélium notamment). Ces souris contrôlescroisées avec des mâles transgéniques développentune hypertension et une protéinurie gestationnelles. L’hypertension est aussi élevé que celleobservée chez des patientes atteintes de prééclampsie sévère. Les reins de ces souris ont des anomaliesqui rappellent la pathologie humaine, avec desdépôts de fibrine. Enfin, nous avons testé un traitement : l’aspirine à faible dose qui est efficace pourempêcher l’hypertension et limite les altérationsrénales et la protéinurie. Nous avons obtenu unmodèle unique pour étudier la prééclampsiesévère, qui indique sans aucun doute un rôle clé deSTOX1 dans la physiopathologie. Cet outil serautile pour trouver de nouvelles voies qui pourraientêtre impliquées dans la physiopathologie de laprééclampsie et pour tester de nouvelles approchesthérapeutiques.

DNA methylation is an important part of the epigenetic code governing gene expression. In human reproductive diseases, recent studies have shown the existence of deviations from the normal methylation profile at various genome loci. In this review, this type of epigenetic alterations is explored in pathological spermatogenesis, ovarian diseases, placental syndromes, such as preeclampsia and Intra- Uterine Growth Restriction, uterine diseases such as endometriosis, and putative pathophysiological effects of Assisted Reproductive Technologies. We review the notion of epigenetics, the technical methods available to analyze methylation, and the known associations between reproductive diseases and DNA methylation, focusing on human pathologies and on animal models when available. We show that imprinted genes control regions (ICRs) are a prominent and frequent target of methylation anomalies in reproductive disorders, but such alterations also affect non-imprinted genes. The mechanistic aspects of gene regulation in response to methylation anomalies are also discussed in this review when they have been investigated.

Pharmacological interventions in preterm labour attempt to target various protagonists involved in the signalling pathway leading to myometrial contractions. None of these interventions has clearly demonstrated any real clinical benefit, and some are associated with severe maternal and fetal side effects. The original study now reported has disclosed a new potential target, myometrial phosphodiesterase-4 (PDE4), for tocolysis. The PDE4 family specifically hydrolyses cAMP, a major intracellular second messenger implicated in the mechanisms governing smooth muscle motility. One particular isoform, PDE4B2, is overexpressed in the human myometrium at the end of pregnancy, and we observed a change of the myorelaxant properties of selective PDE4 inhibitors at this time. Signalling factors involved in labour at term or in infection-induced preterm labour, such as PGE 2 or IL-1β, are also able to induce PDE4B2 expression by way of a cAMP-dependent pathway. This feedback loop explains, at least in part, the process of desensitisation to relaxant agents that occurs at the end of pregnancy. In addition, PDE4 inhibitors block the production of LPS-induced inflammatory cytokines by myometrial explants. Thus, a combination of myorelaxant and anti-inflammatory properties make the PDE4 inhibitors particularly attractive as a target for the prevention of threatened infection-induced preterm delivery and its consequences for the premature infant.

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International audience; Chorioamnionitis is implicated in the pathophysiology of bronchopulmonary disease, and the associated inflammatory response is responsible for adverse effects on alveolar development. The aim of this work was to analyze the effects of a phosphodiesterase 4 (PDE4)-selective inhibitor, rolipram (a modulator of the inflammatory response), in an experimental model of chorioamnionitis on pulmonary development and on the processes of infection and inflammation. Rabbit mothers were assigned to four groups: 1) saline serum inoculation (controls); 2) Escherichia coli intrauterine inoculation (C+); 3) rolipram infusion (R+); and 4) E. coli inoculation + rolipram infusion (C+R+). High rates of morbility and mortality were noticed in mothers and pups (5 of 13 pregnant rabbits in groups with rolipram). Alveolar development, inflammation, and infection were analyzed in pups at day 0 and day 5. At day 0, in the context of chorioamnionitis, rolipram significantly decreased birth weight (p < 0.01) relative to that of controls (p < 0.05). At day 5, weight normalized in group C+R+ but not in group C+ relative to controls (p < 0.001); moreover, alveolar airspace volume was preserved in group C+R+ but not in group C+ (p < 0.05). Interstitial volume decreased in group C+ versus controls (p < 0.05) but was preserved in group C+R+. Specific alveolar area was not significantly modified by rolipram. No significant difference was found concerning bronchoalveolar lavage cellularity, and all blood cultures remained sterile. In this model of impaired alveologenesis, rolipram significantly preserved specific alveolar density. However, PDE4 inhibition induced antenatal fetal demise and growth retardation.