Clues for Improving the Pathophysiology Knowledge for Endometriosis Using Serum Micro-RNA Expression

The pathophysiology of endometriosis remains poorly understood. The aim of the present study was to investigate functions and pathways associated with the various miRNAs differentially expressed in patients with endometriosis. Plasma samples of the 200 patients from the prospective “ENDO-miRNA” study were analyzed and all known human miRNAs were sequenced. For each miRNA, sensitivity, specificity, and ROC AUC values were calculated for the diagnosis of endometriosis. miRNAs with an AUC ≥ 0.6 were selected for further analysis. A comprehensive review of recent articles from the PubMed, Clinical Trials.gov, Cochrane Library, and Web of Science databases was performed to identify functions and pathways associated with the selected miRNAs. In total, 2633 miRNAs were found in the patients with endometriosis. Among the 57 miRNAs with an AUC ≥ 0.6: 20 had never been reported before; one (miR-124-3p) had previously been observed in endometriosis; and the remaining 36 had been reported in benign and malignant disorders. miR-124-3p is involved in ectopic endometrial cell proliferation and invasion and plays a role in the following pathways: mTOR, STAT3, PI3K/Akt, NF-κB, ERK, PLGF-ROS, FGF2-FGFR, MAPK, GSK3B/β–catenin. Most of the remaining 36 miRNAs are involved in carcinogenesis through cell proliferation, apoptosis, and invasion. The three main pathways involved are Wnt/β–catenin, PI3K/Akt, and NF–KB. Our results provide evidence of the relation between the miRNA profiles of patients with endometriosis and various signaling pathways implicated in its pathophysiology.

Model of transfection in human endometrial epithelium cells with hsa-miR-191-5p Model of transfection in human endometrial epithelium cells with hsa-miR-191-5p

Background: Human endometrial cells are important in blastocyst recognition and implantation. We have recently shown that miR-191-5p secreted into culture medium by human embryos cultured and transferred to woman on the fifth day of development was associated with the percentage of pregnant vs. non-pregnant patients. Little is known about the regulation and expression of endometrial miRNAs induced by embryonic miRNAs in endometrial tissue. Therefore, in the present work we explored the viability and transfection of RL95-2 endometrial cell line with agomiR-191. Results: The main results obtained in this study were: First, transfection of RL95-2 cell line with 100nM of lipofectamine in combination with 15, 30, and 60 nM of agomiR-191 for 3, 6 and 24 hours does not affect the viability of RL95-2 cells. Second, we observed expression of miR-191 with 60 pmol of agomiR-191 in a time dependent transfection. Conclusion:: Stimulation of RL95-2 endometrial cell line with lipofectamine does not modify their viability. The transfected RL95-2 endometrial cells showed increased the expression of miR-191.

AG-205 Upregulates Enzymes Involved in Cholesterol Biosynthesis and Steroidogenesis in Human Endometrial Cells Independently of PGRMC1 and Related MAPR Proteins

An inappropriate response to progestogens in the human endometrium can result in fertility issues and jeopardize progestin-based treatments against pathologies such as endometriosis. PGRMC1 can mediate progesterone response in the breast and ovaries but its endometrial functions remain unknown. AG-205 is an alleged PGRMC1 inhibitor but its specificity was recently questioned. We added AG-205 in the cultures of two endometrial cell lines and performed a transcriptomic comparison. AG-205 significantly increased expression of genes coding enzymes of the cholesterol biosynthetic pathway or of steroidogenesis. However, these observations were not reproduced with cells transfected with siRNA against PGRMC1 or its related proteins (MAPRs). Furthermore, AG-205 retained its ability to increase expression of selected target genes even when expression of PGRMC1 or all MAPRs was concomitantly downregulated, indicating that neither PGRMC1 nor any MAPR is required to mediate AG-205 effect. In conclusion, although AG-205 has attractive effects encouraging its use to develop therapeutic strategies, for instance against breast cancer, our study delivers two important warning messages. First, AG-205 is not specific for PGRMC1 or other MAPRs and its mechanisms of action remain unclear. Second, due to its effects on genes involved in steroidogenesis, its use may increase the risk for endometrial pathologies resulting from imbalanced hormones concentrations.

Zinc(II) niflumato complex effects on MMP activity and gene expression in human endometrial cell lines

Endometriosis is a chronic inflammatory disease which increasingly affects young women under 35 years of age and leads to subfertility even infertility. Analysis of the cytotoxic effect of zinc(II) niflumato complex with neocuproine ([Zn(neo)(nif)2] or Zn-Nif) on immortalized human endometriotic cell line (12Z) and on control immortalized human endometrial stromal cell line (hTERT) was performed using xCELLigence technology for approximately 72 h following the treatment with Zn-Nif as well as cell viability Trypan Blue Assay. 12Z cell line proliferated more slowly compared to unaffected cells, whereas hTERT cells did not show similar behavior after treatment. The complex probably reduces the effect of pro-inflammatory pathways due to the effect of NSAID, while presence of zinc might reduce the level of ROS and regulate ER2 levels and MMP activity. The observed effects and high selectivity for rapidly proliferating cells with increased inflammatory activity suggest a good prognosis of successful decrease of endometriosis stage with this complex.

O-046 Implantation and beyond, the endometrial perspective

text Early pregnancy failures are spread throughout the first few weeks, suggesting that impairment of normal biological processes can occur at sequential stages: epithelial attachment and breaching, early stromal and then decidual invasion, glandular and vascular invasion. Biopsy-based transcriptomics and proteomics have failed to demonstrate a reproducible or interpretable molecular signature for endometrial receptivity, but single cell RNAseq suggests a step change in the epithelial transcriptome in the mid secretory phase, consistent with the appearance of new cell phenotype(s). Previously unseen heterogeneity in both epithelial and stromal cell populations has become evident, but gene signatures have not yet achieved a level of resolution that allows insights clear enough to decode the biology of the receptive state. However, methodology for propagating and recombining endometrial cell populations into 3D engineered tissue models has advanced, so that new mechanistic questions can be asked. Initial acquisition of receptivity to implantation is followed by the development of a supportive environment for embryos that possess the capacity to progress, with maternal cell populations (epithelial, stromal, immune and vascular) acting cooperatively within a remodelling extracellular matrix (ECM). A balance must be achieved in the ECM between breakdown, with opening of hydrated spaces to allow expansion of the embryonic sac while maintaining a substrate for stable physical attachment to allow invasion by extravillous trophoblast. The extent and nature of cellular trafficking to and from the uterus is important before and during early pregnancy. There is evidence that regulation of a resident senescent cell subpopulation by uterine NK cells occurs in concert with ECM remodelling to achieve a functionally supportive environment in the early first trimester. Access of bone marrow-derived cells to vessel walls is regulated by placental-endothelial signalling to initiate remodelling for the increased blood flow to the conceptus that is required in later pregnancy. Thus ‘receptivity’ and ‘supportiveness’ require normal cell proportions and functional phenotypes in multiple endometrial cell populations and their physical environment in order to allow a well-calibrated sequential developmental response in the conceptus.

Modulation of Bovine Endometrial Cell Receptors and Signaling Pathways as a Nanotherapeutic Exploration against Dairy Cow Postpartum Endometritis

In order to control and prevent bovine endometritis, there is a need to understand the molecular pathogenesis of the infectious disease. Bovine endometrium is usually invaded by a massive mobilization of microorganisms, especially bacteria, during postpartum dairy cows. Several reports have implicated the Gram-negative bacteria in the pathogenesis of bovine endometritis, with information dearth on the potentials of Gram-positive bacteria and their endotoxins. The invasive bacteria and their ligands pass through cellular receptors such as TLRs, NLRs, and biomolecular proteins of cells activate the specific receptors, which spontaneously stimulates cellular signaling pathways like MAPK, NF-kB and sequentially triggers upregulation of pro-inflammatory cytokines. The cascade of inflammatory induction involves a dual signaling pathway; the transcription factor NF-κB is released from its inhibitory molecule and can bind to various inflammatory genes promoter. The MAPK pathways are concomitantly activated, leading to specific phosphorylation of the NF-κB. The provision of detailed information on the molecular pathomechanism of bovine endometritis with the interaction between host endometrial cells and invasive bacteria in this review would widen the gap of exploring the potential of receptors and signal transduction pathways in nanotechnology-based drug delivery system. The nanotherapeutic discovery of endometrial cell receptors, signal transduction pathway, and cell biomolecules inhibitors could be developed for strategic inhibition of infectious signals at the various cell receptors and signal transduction levels, interfering on transcription factors activation and pro-inflammatory cytokines and genes expression, which may significantly protect endometrium against postpartum microbial invasion.

Scar endometriosis diagnosed as incisional hernia before surgery

Scar endometriosis is a rare condition highly related to history of abdominal surgery. Due to the low incidence, it is often misdiagnosed. A woman presented to the surgery outpatient clinic with a mass near her C-section scar. Physical examination and ultrasound suggested Incisional Hernia while intraoperative finding revealed a mass suggestive of endometriosis which later confirmed by pathology examination. Scar endometriosis is a common subtype of extra-pelvic endometriosis. Iatrogenic transplantation is speculated to be its etiopathogenesis. Preoperatively, it is challenging to differentiate endometriosis from another abdominal masses. The definitive diagnosis is based on laparoscopy or surgery with histological verification. Chronic pain is complex and often involves multiple factors beyond simply a diagnosis of endometriosis, but it is important to think of endometriosis on women patients presenting with a mass and cyclic pain with history of surgery involving a large amount of endometrial cell.