Decidualization of endometrial stromal cells is a critical process for establishing uterine receptivity and successful embryo implantation, involving coordinated biochemical, cellular, and biomechanical remodeling. Emerging evidence indicates that this process is associated with activation of endoplasmic reticulum stress (ERS) and the development of distinct stromal subpopulations, including mature and senescent decidual cells. However, how these pathways integrate to regulate the mechanical properties of the endometrium remains unclear. Here, we investigated the impact of decidualization dysregulation on stromal cell nanomechanics and its contribution to implantation failure. Our first approach used an in vitro decidualization model of human endometrial stromal cells (cell line), in which an excessive ERS was induced by thapsigargine. Then, decidualization, senescence and unfolded protein response were evaluated by molecular analyses, cellular nanomechanical properties by atomic force microscopy, and decidual functionality with a blastocyst-like spheroid implantation assay (using Swan71 cells). Decidualization induced a significant decrease in cellular stiffness (Young's modulus), consistent with a more permissive phenotype for embryo implantation. In contrast, exacerbated ERS-induced prior to decidualization-disrupted the balance between mature and senescent decidual cells and prevented this biomechanical softening, restoring stiffness to levels comparable to those on nondecidualized cells. Functionally, these alterations impaired trophoblast adhesion and expansion in vitro. Finally, to provide a physiologically relevant view of how decidualization operates in vivo, we studied endometrial samples from fertile women (N = 13) and from women with recurrent implantation failure (N = 11). These patients exhibited reduced expression of key decidualization and senescence markers evaluated by qPCR, similar to our results with exacerbated ERS, supporting the clinical relevance of our findings. Together, our results demonstrate that successful decidualization requires tight coordination between ERS, cellular senescence, and biomechanical remodeling. Disruption of this integrated program leads to a mechanically nonpermissive endometrium, providing a novel mechanistic framework for implantation failure.

Background: Background: Endometriosis affects 10% of women of reproductive age. Despite the well-known association between endometriosis and infertility, the mechanisms underlying this association remain to be elucidated. Methods: Implantation and pregnancy success rates were evaluated by a retrospective study of patients that underwent IVF using euploid embryos comparing healthy vs. endometriosis patients. To study the early embryo–endometrial dialogue, an interactome network was constructed using public RNAseq data from normal secretory-phase endometrial samples and day-5 blastocyst. Public bulk and single-cell RNAseq data from endometrial samples of endometriosis patients were used to detect alterations in the interactome. Results: Endometriosis patients required significantly more IVF attempts compared to those without endometrial pathologies; however, once pregnancy was achieved, the evolution of both groups was similar. The interactome network between normal endometrium and day-5 blastocyst showed a significant enrichment of pathways associated with tissue remodelling, angiogenesis, and immune regulation, which were altered in endometriosis patients. Endometriosis patients also presented an increased frequency and activation of NK, CD4+, and CD8+ cells, which interfere with embryo–endometrial dialogue. Conclusions: We identified key molecular processes affected by endometriosis specifically involved in the early interaction between the blastocyst, decidual, and resident immune cells, that may underline the reported fertility problems associated with endometriosis.

Study question Is ovarian aging associated with the generation of inflammatory macrophages with a dysregulated immunometabolism and be ameliorated after treatment with platelet-rich plasma-releasate (PRPr)? Summary answer Aged follicular macrophages show an inflammatory profile associated with an altered metabolism that can be reprogrammed with an alternative therapeutic treatment. What is known already Inflammaging is a systemic and chronic inflammatory response, linked with advanced age. Particularly in the ovary, inflammaging leads to a decrease in the ovarian reserve, oocyte quality and infertility. PRPr therapy is being widely explored to improve ovarian function and increase the chances of pregnancy. Here, we investigate the mechanisms underlying the exacerbated inflammatory response during ovarian aging, focusing on the metabolic profile of follicular macrophages and a novel T lymphocyte regulatory subpopulation, CD3+CD4-CD8- (DNT), that is emerging as effector cells capable of mediating immune tolerance in the female reproductive system. Study design, size, duration Macrophages and lymphocytes were recovered from follicular fluid (FF) from women of different ages and tested by immune-staining and FACS analysis. In macrophages (CD68+), we evaluated lipid accumulation, reactive oxygen species (ROS), lactate and IL-1β production, and mitochondrial membrane potential. Immune cells profiles were correlated with clinic parameters such as number of M2 oocytes and atretic follicles. Finally, macrophages were treated in vitro with PRPr and, after 5 days, reprogramming of macrophages profile was tested. Participants/materials, setting, methods We studied FF (n = 102) from women from ovodonation (21-29y) and ART indication (36-42y). The San Isidro Ethics Committee, Buenos Aires, Argentina, approved this study. Mononuclear cells were isolated using Ficoll-Hypaque and adherent macrophages profile (CD68+ IL-1β+) and DNT cell subpopulation (CD3+CD4-CD8-) tested by flow cytometry. Lipid droplets accumulation, ROS and mitochondrial potential were measured using their respective probes. Follicular macrophages were treated with PRPr (obtained from fertile women) for 5 days. Main results and the role of chance FF macrophages from aged women (35-42y) showed a significant increase in IL-1β production in comparison with the youngest group (21-29y), that also correlates negatively with the number of M2 oocytes recovered. Focusing on macrophage (CD68+) metabolic profile we found an increase in lipid droplets in follicular macrophages from aged women in comparison with the younger group. Moreover, these macrophages had hyperpolarization of the mitochondrial membrane and, as we expected, a higher ROS and lactate production. This leads us to infer that these M1 macrophages undergo glycolysis. Then, the isolated follicular macrophages from aged women were treated with PRPr from fertile women to evaluate its modulatory effects and we observed a reduction in the frequency of CD68+IL-1β+ cells in comparison with those cultured without treatment. Also, treated macrophages showed a reduction in the levels of ROS and lactate production. Finally, we focused on a novel DNT lymphocyte regulatory subpopulation, CD3+CD56-CD4-CD8-, that is emerging as effector cells capable of mediating immunoregulation in the female reproductive system. We found that FF from aged women displayed a reduced frequency of DNT, in comparison with FF from younger women. Limitations, reasons for caution The present results were obtained using FF samples from women from different ages, using a pool of cells from all the follicles in each patient. Even though the samples represent the immune ovarian microenvironment, further studies are necessary to elucidate whether these mechanisms and PRPr treatment operate similarly in vivo. Wider implications of the findings We demonstrated that, during aging, follicular macrophages acquired an altered metabolism related to pro-inflammatory profile, accompanied by a reduction of DNT cells, impacting in oocyte quality. Notably, we showed for the first time that in vitro PRPr treatment reprogrammed macrophage’s profile, suggesting its relevance as a potential therapeutic treatment. Trial registration number No

Endometrial stromal cells acquire a secretory profile associated with endoplasmic reticulum stress (ERS) and unfolded protein response (UPR) related to the onset of a sterile inflammatory response essential for sustaining embryo implantation. However, exacerbated stromal ERS/UPR is associated with reproductive complications. Given the ability of dendritic cells (DCs) to sense stress signals and be conditioned by stromal cells, here we investigated the transmission of ERS (TERS) from stromal cells to monocytes and its impact on tolerogenic DCs conditioning. Blood monocytes were differentiated into DCs (rhGM-CSF+rhIL-4, 5 d) in the presence or absence of conditioned media derived from either thapsigargin-treated (stressed) or nonstressed human endometrial stromal cell line. Soluble factors released by stressed stromal cells impaired CD1a+CD14- DC differentiation and induced a proinflammatory profile, increasing the CD86high cell population, COX-2 expression, and tumor necrosis factor (TNF)-α, interleukin (IL)-8 and IL-1β secretion. Additionally, TERS was observed in these cultures, with increased expression of IRE1α, PERK, and ATF4. Even the splicing of the adaptive UPR marker XBP1 was increased though at low levels, its nuclear translocation was unchanged. These effects on spliced XBP1, coupled with a decreased GRP78/BiP and heightened CHOP expression, suggest the triggering of terminal UPR over adaptive UPR, confirmed by the induction of lytic cell death in stressed cultures. Finally, exacerbated TERS negatively impacted trophoblast migration in a blastocyst-like spheroid in vitro model. These findings suggest that exacerbated stromal ERS can be transmitted to monocytes, altering their differentiation, immune profile, and viability, which could ultimately impair trophoblast migration.

Embryo implantation requires a tight immune homeostatic control that activates regulatory circuits. One interlocutor is the embryo, which produces soluble ligands and expresses receptors for different autocrine and paracrine factors. The other interlocutor is the receptive endometrium, which produces mediators to regulate proliferation, differentiation, adhesion and invasiveness of the embryo, among other processes. Here, we are going to discuss experimental evidence regarding human embryo-endometrial dialogue and give a new insight of the relevance of the immune cells to coordinate embryo implantation. In this sense, decidualization of endometrial stromal cells is a multistep process that gives rise to mature decidual cells and senescent decidual cells. The first subpopulation secretes pro-implantation factors and begins migration by encapsulating the embryo. In turn, the second does not complete differentiation but rather suffers a process of premature senescence that is characterized by the production of pro-inflammatory factors (SASP, senescence-associated secretory phenotype), which contribute to embryo implantation. However, alterations in these processes or in their regulation through microRNAs lead to the perpetuation of an inflammatory response and alterations in endometrial receptivity. Considering that decidual cells acquire the ability to differentially respond to embryo quality, here we also explored how the soluble factors produced by embryos (classified according to their quality) impact on the inflammatory response and shape dendritic and other immune cell recruitment during the peri implantation period. To address these aspects, we present experimental evidence that links endoplasmic reticulum stress, senescence and inflammation and we discuss whether embryos reprogram the immune response.

Ovarian aging is associated with fibro-inflammation, contributing to the decline in oocyte count and quality. Given the immunomodulatory properties of the vasoactive intestinal peptide (VIP) in the reproductive tract, we investigated its role in maintaining ovarian immune homeostasis and preventing premature aging. We evaluated young VIP knockout (KO) mice, comparing them to young wild type (WT) females, for signs of premature aging. Histological staining revealed aberrant ovarian morphology in VIP KO mice, characterized by increased atretic follicles and decreased ovarian reserve compared to WT controls. Moreover, VIP KO ovaries showed reduced vascularization, increased collagen deposition and elevated ROS and IL-1β levels. Foamy macrophages were significantly predominant, indicating premature aging in young VIP KO ovaries. To determine potential mechanisms behind these pathogenic changes, we conditioned peritoneal macrophages from young WT or VIP KO mice in vitro with ovarian-conditioned media from young WT or VIP KO mice to mimic the respective ovarian microenvironment. When WT or VIP KO peritoneal macrophages were conditioned with ovarian media from their respective genotypes, lipid droplet accumulation increased compared to control medium. In cross-genotype experiments, WT macrophages conditioned with media from VIP KO ovaries selectively accumulated higher levels of lipid droplets, whereas no differences were observed in VIP KO macrophages conditioned with WT ovarian media. This suggests that VIP KO macrophages are uniquely sensitized to the inflammatory environment of VIP KO ovaries, implicating both ovarian factors and macrophage status. These findings highlight the role of VIP in preventing fibro-inflammation, thereby preserving ovarian health and preventing premature aging.

PROBLEM:The decidualization process conditions monocytes to the immunosuppressive and tolerogenic dendritic cell (DC)-10 profile, a DC subset with high IL-10 production. Since the implantation process implies an embryo-endometrium-immune crosstalk, here we focused on the ability of embryonic soluble factors to modify decidual DC conditioning accordingly with its quality. METHOD OF STUDY:Human endometrial stromal cell line (HESC) decidualized with medroxyprogesterone and dibutyryl-cAMP (Dec) was stimulated with human embryo-conditioned media (ECM), classified as normal (ND) or impaired developed (ID) for 48 h (n = 18/group). Monocytes isolated from six healthy women were differentiated to DCs with rhGM-CSF+rhIL-4 in the presence/absence of conditioned media (CM) from decidualized cells stimulated with ECM or nontreated. RESULTS:We found that decidualized cells stimulated with ECM sustain a myeloid regulatory cell profile on monocyte-derived culture with increased frequency of CD1a-CD14+ and CD83+CD86low cells. ND-Dec sustained the higher expression of the DC-10 markers, HLA-G and IL-10 whereas ID-Dec diminished IL-10 production (ID-Dec: 135 ± 37.4 vs. Dec: 223.3 ± 49.9 pg/mL, p < 0.05). The treatment with ECM-Dec sustained a higher IL-10 production and prevented the increase of CD83/CD86 after LPS challenge regardless of embryo quality. Notably, TNF-α production increased in ID-Dec cultures (ID-Dec: 475.1 ± 134.7 vs. Dec: 347.5 ± 98 pg/mL, p < 0.05). CONCLUSIONS:Although remaining in a tolerogenic profile compatible with DC-10, DCs can differentially respond to decidual secreted factors based on embryo quality, changing their secretome. These results suggest that in the presence of arrested embryo, DCs could differentially shape the immunological microenvironment, contributing to arrested embryo clearance during the menstrual phase.

OBJECTIVE:To evaluate factors involved in spontaneous pregnancy rate after surgery for endometriosis in patients with endometriosis and infertility.METHODS:This retrospective study spanned from 2014 to 2020 and included a follow-up period of two years of patients with endometriosis-related infertility who underwent laparoscopic surgery. Women aged 25 to 43 years with patent tubes, no/mild male factor and no other infertility factors were selected and grouped according to fertility management as follows: patients immediately prescribed ART (16.5%, ART-p); patients who chose not to undergo ART (83.5%) and achieved spontaneous pregnancy (71.8% SP-p); and patients who first chose not to undergo ART but had it subsequently (28.2%, NSP-p).RESULTS:A total of 200 patients were analyzed. Of the 167 patients who waited for spontaneous pregnancy, 71.8% achieved it. We observed a tendency of higher endometriosis ASRM scores in the ART-p group compared with patients who waited for spontaneous pregnancy, and lower scores in individuals that achieved spontaneous pregnancy. When we looked at how long it took to achieve pregnancy, we found that individuals in the SP-p group achieved pregnancy in 5.7 months, while subjects in the NSP-p group took 1.8 times longer than their peers in the SP-p group (p<0.001). However, once prescribed ART, the individuals in the NSP-p group achieved pregnancy within a similar time when compared with subjects in the SP-p group. In order to identify individuals that might benefit from ART early on, we performed a multivariable analysis and developed a decision tree (81.3% accuracy and 53.3% sensitivity).CONCLUSIONS:The present results indicated that, after surgery, the majority of patients achieved spontaneous pregnancy. The decision tree proposed in this study allows the early identification of patients who might require ART, thus decreasing the time between surgery and pregnancy and improving overall outcomes.

Abstract Background Placental macrophages, Hofbauer cells (HBC) are the only fetal immune cell population within the stroma of healthy placenta along pregnancy. They are central players in maintaining immune tolerance during pregnancy. Immunometabolism emerged a few years ago as a new field that integrates cellular metabolism with immune responses, however, the immunometabolism of HBC has not been explored yet. Here we studied the sex-specific differences in the phenotypic, functional and immunometabolic profile of HBC. Methods HBC were isolated from human term placentas (N = 31, 16 from male and 15 female neonates). Ex vivo assays were carried out to assess active metabolic and endoplasmic reticulum stress pathways by flow cytometry, confocal microscopy, gene expression and in silico approaches. Results HBC from female placentas displayed a stronger M2 phenotype accompanied by high rates of efferocytosis majorly sustained on lipid metabolism. On the other hand, male HBC expressed a weaker M2 phenotype with higher glycolytic metabolism. LPS stimulation reinforced the glycolytic metabolism in male but not in female HBC. Physiological endoplasmic reticulum stress activates IRE-1 differently, since its pharmacological inhibition increased lipid mobilization, accumulation and efferocytosis only in female HBC. Moreover, differential sex-associated pathways accompanying the phenotypic and functional profiles of HBC appeared related to the placental villi environment. Conclusions These results support sex-associated effects on the immunometabolism of the HBC and adds another layer of complexity to the intricate maternal-fetal immune interaction. Graphical abstract

Abstract Endometrial stromal cells acquire a secretory profile associated with a physiological endoplasmic reticulum stress (ERS) and the onset of a sterile inflammatory response to sustain embryo implantation. Here we evaluated the impact of endometrial stromal cells on Dendritic Cells (DC) differentiation through ERS-transmission. We used an in vitro model based on differentiated-monocytes from healthy women (with rhGM-CSF+rhIL-4) cultured in the absence/presence of Conditioned Media (CM) from stressed Human Endometrial Stromal Cells (HESC stimulated with Thapsigargin). Those DC cultures differentiated with stressed-HESC showed a lower frequency of CD1a+CD14- accompanied by an increased CD86high cell-population and higher COX2 expression. We also detected an increase in IL-1β secretion and in AnnexinV+/PI+ cell-population, indicating a lytic programmed cell death. The ERS-transmission was confirmed by increased IRE1α expression on DC. In fact, IRE1α inhibitor prevented the conditioning to a pro-inflammatory DC profile. Finally, to investigate the impact of the DC-inflammatory profile on blastocyst implantation ability, we tested trophoblastic spheroids migration. Blastocyst-like spheroids displayed an altered trophoblast migration when were cultured with supernatant from stressed-DC. These results suggest that ERS might be transmitted from stromal cells to DC and its fine balance is required to maintain tolerance for successful embryo implantation and trophoblast migration.

Abstract Ovarian aging is associated with inflammation, contributing to the decline in oocyte count and quality. Since the vasoactive intestinal peptide (VIP) is an immunopeptide found in the reproductive tract with anti inflammatory effects inducing a tolerogenic response, we focus on VIP contribution to ovarian immune homeostasis preventing premature aging. We observed that young VIP KO mice (3 months old) display an altered ovarian histology with an increase in atretic follicles and less mature follicles as in aged WT mice (8 months old). This microenvironment was accompanied by an increased fibrosis deposition, ROS and IL-1β production. Interestingly, we found a significant decrease in ovarian reserve and the presence of foamy macrophages in young VIP KO ovary associated with premature aging. Getting insight into the mechanism involved in foam cell generation, we pre-conditioned young WT or VIP KO macrophages with WT or VIP KO ovarian media for 48h to mimic the ovarian microenvironment. Macrophages from VIP KO mice display a differential phagocytic ability of apoptotic bodies after preconditioning with WT or VIP KO ovarian media. However, only in VIP KO macrophages conditioned with VIP KO ovarian media we found an increase in lipid droplet accumulation, consistent with the formation of foamy macrophages. These findings highlight the role of VIP in regulating ovarian immune microenvironment, thereby preserving ovarian health and preventing premature aging.

Research question: Do microRNAs (miRNAs) play a role in regulating endoplasmic reticulum stress (ERS) and unfolded protein response (UPR) in decidualized cells and endometrium associated with reproductive failures?Design: Endometrial stromal cell line St-T1b was decidualized in vitro with 8-Br-cAMP over 5 days, or treated with the ERS inducer thapsigargin. Expression of ERS sensors, UPR markers and potential miRNA regulators was analysed by quantitative PCR. Endometrial biopsies from patients with recurrent pregnancy loss (RPL) and recurrent implantation failure (RIF) were investigated for the location of miRNA expression.Results: Decidualization of St-T1b cells resulted in increased expression of ERS sensors including ATF6a, PERK and IRE1a, and the UPR marker, CHOP. TXNIP, which serves as a link between the ERS pathway and inflammation, as well as inflammasome NLRP3 and interleukin 1b expression increased in decidualized cells. An in-silico analysis identified miR-17-5p, miR-21-5p and miR-193b-3p as miRNAs potentially involved in regulation of the ERS/UPR pathways and inflammation associated with embryo implantation. Their expression decreased significantly (P <= 0.0391) in non-decidualized cells in the presence of thapsigargin. Finally, expression of the selected miRNAs was localized by in-situ hybridization in stromal and glandular epithelial cells in endometrial samples from patients with RPL and RIF. Expression in stroma cells from patients with RPL was lower in comparison with stroma cells from patients with RIF.Conclusions: Decidualization in St-T1b cells is accompanied by ERS/UPR processes, associated with an inflammatory response that is potentially influenced by miR-17-5p, miR-21-5p and miR-193b-3p. These miRNAs are expressed differentially in stromal cells from patients with RPL and RIF, indicating an alteration in regulation of the ERS/UPR pathways.

An efficient immune defense against pathogens requires sufficient basal sensing mechanisms that can deliver prompt responses. Type I IFNs are protective against acute viral infections and respond to viral and bacterial infections, but their efficacy depends on constitutive basal activity that promotes the expression of downstream genes known as IFN-stimulated genes (ISGs). Type I IFNs and ISGs are constitutively produced at low quantities and yet exert profound effects essential for numerous physiological processes beyond antiviral and antimicrobial defense, including immunomodulation, cell cycle regulation, cell survival, and cell differentiation. Although the canonical response pathway for type I IFNs has been extensively characterized, less is known regarding the transcriptional regulation of constitutive ISG expression. Zika virus (ZIKV) infection is a major risk for human pregnancy complications and fetal development and depends on an appropriate IFN-β response. However, it is poorly understood how ZIKV, despite an IFN-β response, causes miscarriages. We have uncovered a mechanism for this function specifically in the context of the early antiviral response. Our results demonstrate that IFN regulatory factor (IRF9) is critical in the early response to ZIKV infection in human trophoblast. This function is contingent on IRF9 binding to Twist1. In this signaling cascade, Twist1 was not only a required partner that promotes IRF9 binding to the IFN-stimulated response element but also an upstream regulator that controls basal levels of IRF9. The absence of Twist1 renders human trophoblast cells susceptible to ZIKV infection.

Among 146 potential coding sequences, the most comprehensive eutherian growth hormone gene data set annotated 100 complete coding sequences. The eutherian comparative genomic analysis protocol first described 5 major gene clusters of eutherian growth hormone genes. The present updated gene classification and nomenclature of eutherian growth hormone genes integrated gene annotations, phylogenetic analysis and protein molecular evolution analysis into new framework of future experiments. The curated third party data gene data set of eutherian growth hormone genes was deposited in European Nucleotide Archive under accession numbers LM644135–LM644234.

A better understanding of how the spatial configuration of cities, understood as urban structure and forms, can achieve sustainable development is needed. This paper presents spatial data and an automated workflow for studying the urban structures (i.e., road and transportation networks) and forms (i.e., building size, position, function and density) of two medium-sized European cities - Bergen, Norway and Zürich, Switzerland. The data focuses on examining correlations between the densification patterns and transport energy usage of these cities de Koning et al., (2020). Spatial and tabular datasets for (i) urban structures, (ii) urban forms, (iii) building density, (iv) road centre lines and (v) transport energy usage are obtained as georeferenced files from OpenStreetMap (OSM) and upon request from collaborating local and national authorities. Transport energy data is derived from traffic data collected from the Norwegian Public Road Authorities or simulated via a traffic model. Open-source data is used wherever possible. Data gaps within proprietary data are supplemented with proxies or open-source data.Hand-drawn axial maps drawn by the authors using the Space Syntax methods and analysed via depthmapX software are a crucial dataset presented here. All analysed data are then returned to a Geographical Information System (GIS) platform and processed via an automated workflow of 19 steps built via the ModelBuilderTM tool in ESRI® ArcGIS. The automated workflow allows for repetitive cross-city comparison and the compilation of diverse spatial data sources for analysis.In combination with the novel workflow, the dataset can be used for future comparative studies in spatial planning, transport planning and management of energy systems to facilitate informed decision-making towards more sustainable developments.

BACKGROUND:A tight immune and metabolic regulation underlies the early maternal-placental interaction to assist the energetic dynamic demands of the fetus throughout pregnancy. The vasoactive intestinal peptide (VIP) holds biochemical, metabolic and immune properties consistent with a regulatory role during pregnancy.AIM:Here we overview critical aspects of embryo implantation and placental development with special focus on the immune and metabolic effects of VIP expressed by decidual and trophoblast cells.CONTENT:During decidualization, endometrial stromal cells undergo reticular stress and trigger unfolded protein response (UPR) that enable expansion of their endoplasmic reticulum and immunomodulatory factor synthesis. These processes appear differentially affected in recurrent abortion and in vitro fertilization failure suggesting their relevance in reproductive pathologies. Similarly, defective placentation associates with altered immune, vascular and trophoblast interaction resulting in complicated pregnancies that threaten maternal and neonatal health and underlie metabolic programming of adult life. We discuss the most recent research on decidual, trophoblast and immune cell interaction on the light of VIP regulation. Its role in decidualization and UPR associated with a sterile inflammatory response and angiogenesis is discussed. Evidence on VIP modulation of cytotrophoblast cell function, metabolism and immune profile is revised as well as the shaping of decidual leukocyte phenotype and function from decidualization to term.IMPLICATIONS:The broad spectrum of effects of VIP from implantation to term in normal and pathological conditions summarized here might contribute to the identification of novel biomarkers for diagnosis and pharmacological targeting.

Premature placental aging is implicated in a number of complications of pregnancy including preeclampsia. A placenta knockout mouse model has shown a relationship between SIRT1, aging and placental dysfunction. The role of SIRT1 in cellular senescence has been extensively studied in various cell types, but its role in trophoblast senescence is almost unknown.Human placental samples were obtained from preeclampsia-affected women and healthy controls. The placental aging profiles were assessed by Doppler ultrasound, placental histopathology, and evaluation of senescence- and ECM-related markers. The SIRT1 expression pattern relevant to placental aging profiles was studied in premature aging placenta with preeclampsia (32–37 weeks gestation, n = 10) and healthy controls (37–40 weeks gestation, n = 10). Using cell culture, the effects of activation and knockdown of SIRT1 or its downstream target molecules in syncytialized BeWo cells were evaluated.SIRT1 was expressed by syncytiotrophoblast across normal gestation. In preeclamptic premature aging placentas, SIRT1 was significantly downregulated, while senescence- and extracellular matrix (ECM) -related protein levels were upregulated compared to controls. Immunohistochemistry showed these changes to be confined to syncytiotrophoblast. In vitro, SIRT1 activation in response to resveratrol (RSV) abrogated senescence in forskolin-induced syncytialization of BeWo cells via regulation of senescence- and ECM-related proteins, and filamentous actin (F-actin). These effects were restored by SIRT1 siRNA.The downregulation of SIRT1 may accelerate senescence of syncytiotrophoblast via targets contributing to regulation of the cell cycle, ECM production and cytoskeleton reorganization leading to premature placental aging observed in preeclampsia.

PROBLEM:Decidualized cells display an active role during embryo implantation sensing blastocyst quality, allowing the implantation of normal developed blastocysts and preventing the invasion of impaired developed ones. Here, we characterized the immune microenvironment generated by decidualized cells in response to soluble factors secreted by blastocysts that shape the receptive milieu.METHOD OF STUDY:We used an in vitro model of decidualization based on the Human Endometrial Stromal Cells line (HESC) differentiated with medroxiprogesterone and dibutyryl-cAMP, then treated with human blastocysts-conditioned media (BCM) classified according to their quality.RESULTS:Decidualized cells treated with BCM from impaired developed blastocysts increased IL-1β production. Next, we evaluated the ability of decidualized cells to modulate other mediators associated with menstruation as chemokines. Decidualized cells responded to stimulation with BCM from impaired developed blastocysts increasing CXCL12 expression and CXCL8 secretion. The modulation of these markers was associated with the recruitment and activation of neutrophils, while regulatory T cells recruitment was restrained. These changes were not observed in the presence of BCM from normal developed blastocysts.CONCLUSION:Soluble factors released by impaired developed blastocysts induce an exacerbated inflammatory response associated with neutrophils recruitment and activation, providing new clues to understand the molecular basis of the embryo-endometrial dialogue.