Stanniocalcin-1 expression in normal human endometrium and dysregulation in endometriosis

Objective To determine expression of stanniocalcin-1 (STC1) in human endometrium with and without endometriosis and its regulation by steroid hormones. Design Laboratory study. Setting University. Patient(s) Nineteen women with endometriosis and 33 control women. Intervention(s) Endometrial biopsy and fluid sampling. Main Outcome Measure(s) Analysis of early secretory (ESE) and midsecretory (MSE) endometrial secretomes from fertile women with the use of nano–liquid chromatography–dual mass spectrometry; real-time quantitative polymerase chain reaction, and immunohistochemistry for STC1 and its receptor calcium-sensing receptor (CASR) mRNA and proteins in endometrium with and without endometriosis; evaluation of STC1 and CASR mRNA expression in endometrial stromal fibroblasts (eSF) from women with and without endometriosis decidualized with the use of E2P or 8-bromo-cyclic adenosine monophosphate (cAMP). Result(s) STC1 protein was strongly up-regulated in MSE versus ESE in endometrial fluid of fertile women. STC1 mRNA significantly increased in MSE from women with, but not from those without, endometriosis, compared with proliferative endometrium or ESE, with no significant difference throughout the menstrual cycle between groups. STC1 mRNA in eSF from control women increased >230-fold on decidualization with the use of cAMP versus 45-fold from women with endometriosis, which was not seen on decidualization with E2/P. CASR mRNA did not exhibit significant differences in any condition and was not expressed in isolated eSF. STC1 protein immunoexpression in eSF was significantly lower in women with endometriosis compared with control women. Conclusion(s) STC1 protein is significantly up-regulated in MSE endometrial fluid and is dysregulated in eutopic endometrial tissue from women with endometriosis. It is likely regulated by cAMP and may be involved in the pathogenesis of decidualization defects

Meta-signature of human endometrial receptivity : a meta-analysis and validation study of transcriptomic biomarkers

Previous transcriptome studies of the human endometrium have revealed hundreds of simultaneously up-and down-regulated genes that are involved in endometrial receptivity. However, the overlap between the studies is relatively small, and we are still searching for potential diagnostic biomarkers. Here we perform a meta-analysis of endometrial-receptivity associated genes on 164 endometrial samples (76 from 'pre-receptive' and 88 from mid-secretory, 'receptive' phase endometria) using a robust rank aggregation (RRA) method, followed by enrichment analysis, and regulatory microRNA prediction. We identify a meta-signature of endometrial receptivity involving 57 mRNA genes as putative receptivity markers, where 39 of these we confirm experimentally using RNA-sequencing method in two separate datasets. The meta-signature genes highlight the importance of immune responses, the complement cascade pathway and the involvement of exosomes in mid-secretory endometrial functions. Bioinformatic prediction identifies 348 microRNAs that could regulate 30 endometrial-receptivity associated genes, and we confirm experimentally the decreased expression of 19 microRNAs with 11 corresponding up-regulated meta-signature genes in our validation experiments. The 57 identified meta-signature genes and involved pathways, together with their regulatory microRNAs could serve as promising and sought-after biomarkers of endometrial receptivity, fertility and infertility.Peer reviewe

Endometrial, embryonic and ovarian aspects of human implantation

The general aim of the studies was to add more understanding to the mechanism of human embryo implantation, which will allow us to improve pregnancy rates in assisted reproduction. The simultaneous positive spatial and temporal expression of pinopodes with LIF, LIFR and HBEGF proteins in endometrial samples from healthy women was found. LIF and LIFR had different expression patterns. HB-EGF was present both inside the luminal epithelial cells and on the surface of pinopodes. The findings suggest that the simultaneous molecular and structural cell changes are important in the initiation of human blastocyst implantation. Human embryonic stem cells (hESC), derived from the inner cell mass (ICM) of human blastocyst, provide an excellent model for studying the events occurring in the embryonic side of the implantation. Gene expression in the hESC line HS237 was analyzed using microarrays. Real-time RT-PCR was used to validate the microarray results in four cell lines (HS181, HS235, HS237, HS293). The expression of LIF, LIFR and gpl30 mRNA and protein was increased in differentiated HS237 cells when compared with those in undifferentiated cells. An inhibitor of LIF-mediated signaling, SOCS-1, was up-regulated in undifferentiated hESCs compared with differentiated ones. Increased levels of SOCS-1 might prevent LIF-induced STAT3 signal transduction in undifferentiated hESC and explain the LIF resistance in those cells. Genes, expressed specifically and those shared in undifferentiated hESCs, differentiated hESCs and in fibroblasts were identified. Differentiation of hESC into other cell types began before the changes could be observed in light microscopy, as revealed by scanning electron microscopy. Both hypothyroidism and hyperthyroidism affect menstrual cycle. The mechanisms behind these reproductive abnormalities are not well known. We described the presence and cellular distribution of TSH and thyroid hormone receptors (TR) protein and mRNA in normal human endometrium throughout the menstrual cycle and in human ovary. After stimulation with TSH and thyroid hormone, endometrial epithelial and stromal cells showed increased cAMP (activation of TSHR) and ERK1/2 (activation of TR receptors) expression. Significant down-regulation of LIF gene under thyroid hormone treatment in endometrial cells suggested involvement of thyroid hormone in human implantation. Granulosa cells (M) stimulated with TSH showed a significant increase in cAMP production, indicating activation of the TSHR. Stimulation with T4 resulted in increased concentration of ERK1/2 (activation of TR receptors). Real-time PCR did not show significant changes in the expression of glucose transporter1 and estrogen receptors (ER) alpha in endometrial cells or ERbeta genes in ovarian tissue or GCs after stimulation with TSH or T4. The expression of TSHR, TRalpha1 and TRbeta1 receptors in the human ovary and endometrium suggests that reproductive disorders during thyroid dysfunction might result from direct effects of thyroid hormones on the ovaries and endometrium. Their strong expression in ovarian surface epithelium suggests a particular role in those cells, but the exact role remains to be solved

Making More Womb: Clinical Perspectives Supporting the Development and Utilization of Mesenchymal Stem Cell Therapy for Endometrial Regeneration and Infertility

The uterus is a homeostatic organ, unwavering in the setting of monthly endometrial turnover, placental invasion, and parturition. In response to ovarian steroid hormones, the endometrium autologously prepares for embryo implantation and in its absence will shed and regenerate. Dysfunctional endometrial repair and regeneration may present clinically with infertility and abnormal menses. Asherman’s syndrome is characterized by intrauterine adhesions and atrophic endometrium, which often impacts fertility. Clinical management of infertility associated with abnormal endometrium represents a significant challenge. Endometrial mesenchymal stem cells (MSC) occupy a perivascular niche and contain regenerative and immunomodulatory properties. Given these characteristics, mesenchymal stem cells of endometrial and non-endometrial origin (bone marrow, adipose, placental) have been investigated for therapeutic purposes. Local administration of human MSC in animal models of endometrial injury reduces collagen deposition, improves angiogenesis, decreases inflammation, and improves fertility. Small clinical studies of autologous MSC administration in infertile women with Asherman’s Syndrome suggested their potential to restore endometrial function as evidenced by increased endometrial thickness, decreased adhesions, and fertility. The objective of this review is to highlight translational and clinical studies investigating the use of MSC for endometrial dysfunction and infertility and to summarize the current state of the art in this promising area

The Effect of Human Growth Hormone on Endometrial Growth in Controlled Ovarian Hyperstimulation Cycles

This study aims to compare endometrial growth before and after the addition of human growth hormone (hGH) in controlled ovarian hyperstimulation (COH) cycles. A 5-year retrospective cohort study of patients treated with hGH to improve oocyte development during COH cycles was conducted. Each patient’s cycle without hGH immediately preceding cycle(s) with hGH was used for patients to serve as their own controls. Primary outcome was absolute growth in endometrial thickness from pre-stimulation start to day of hCG trigger. Mixed-model regression analysis controlled for patient correlation over repeat cycles and potential confounders. 80 patients were included. Mean age was 39.7 years; mean BMI was 23.8 kg/m2. Majority of patients were nulliparous, non-smoking, and White or Asian. Most common diagnosis was diminished ovarian reserve. Endometrial growth was compared between 159 COH cycles with hGH and 80 COH control cycles; mean increase was 4.5 mm and 3.9 mm, respectively-an unadjusted difference of 0.6 mm (95% CI: 0.2–1.1, p = 0.01). After adjusting for demographic/clinical factors, hGH was associated with 0.9 mm greater endometrial growth (0.4–1.4, p < 0.01). Absolute increase in endometrial thickness was higher in COH cycles that included hGH. Further prospective studies in embryo transfer cycles are needed