Project description:Purpose: This study was undertaken to evaluate the effect of peritoneal fluid from subjects with differing stages of endometriosis on gene expression in endometrial stromal cells. Methods: Peritoneal fluid from subjects with minimal, moderate, and severe stages of endometriosis or without endometriosis (controls) was collected, filtered and separated from peritoneal cells. Telomerase – immortalized human endometrial stromal cells (T-HESC) were treated with the peritoneal fluid samples for 48 hours. RNA was isolated from treated cells and processed for DNA microarray. Results: 162 genes were found to be differentially expressed in T-HESC cells treated with peritoneal fluid from subjects with differing stages of endometriosis. Three of the differentially expressed genes were chosen for confirmatory studies using quantitative RT-PCR; these were interleukin 8, corin and matrix metalloproteinase 3. The primary gene ontologies identified by microarray highlight functions expected to be involved in the establishment of endometriosis, such as proteases, signal transduction, defense and immune system, cell adhesion, cell cycle and cell death, cytoskeleton, transcriptional regulation and translation, extracellular matrix, and enzymes and metabolism. Conclusions: This study demonstrated that the severity of endometriosis affected the factors present in peritoneal fluid which, in turn, affected gene expression in endometrial stromal cells.
Project description:Endometriosis is an inflammatory disease and bone marrow-derived cells are abundant in endometriotic lesions and in the peritoneal fluid of women with the disease. This study tested the hypothesis that reciprocal communication occurs between macrophages and cultured human endometrial stromal cells and that this communication contributes to the pathology of endometriosis. Changes in gene expression elicited by exposure to factors secreted by the opposing cell type were measured by DNA microarray to test this hypothesis. 716 named genes were differentially expressed in cultured endometrial stromal cells in response to factors secreted by macrophages. Genes that were up-regulated included IL8/CXCL8, MMP3, phospholamban, CYR61/CCN1, CTGF/CCN2, tenascin C, and NNMT, whereas integrin alpha 6 was down-regulated. In contrast, 15 named genes were differentially expressed in macrophages in response to factors secreted by cultured endometrial stromal cells. The data document reciprocal communication between macrophages and endometrial stromal cells and suggest that interaction with macrophages stimulates the expression of genes in endometrial stromal cells that contribute to migration, adhesion, invasion, neovascularization and mitosis of endometrial cells that may support the establishment of endometriosis.
Project description:We primary cultured ectopic endometrial cells from patients with endometriosis (2 cases) and without endometriosis(2 cases) and collected cell culture supernatants(P0). We isolated exosomes from cell culture supernatants by differential centrifugation and then performed proteome analysis on the two groups of exosomes to investigate the role of ectopic endometrial cell-derived exosomes in the development of endometriosis.
Project description:The hypothesis that male michrochimerism in eutopic endometrium is a factor for endometriosis, as indicated by indirect evidence was examined in endometrial samples from control (Group 1) and stage IV ovarian endometriosis (Group 2), either fertile (Group 1A and 2A) or Infertile (Group 1B and 2B) pateints. 6 coding and 10 non-coding genes showed bi-modal pattern of expression characterised by low expression in samples obtained from fertile patients and high expressions in infertile patients. Several coding and non-coding MSY-linked genes displayed michrochimerism in form of presence of their respective DNA inserts along with their microarray-detectable expression in endometrium irrespective of fertility history and disease.
Project description:Background. Endometriosis is a common, complex disorder which is underrecognized and subject to prolonged delays in diagnosis. It is accompanied by significant changes in the eutopic endometrial lining. Methods . We have undertaken the first single cell RNA-sequencing (scRNA-Seq) comparison of endometrial tissues in freshly collected menstrual effluent (ME) from 33 subjects, including confirmed endometriosis patients (cases) and controls as well as symptomatic subjects (who have chronic symptoms of endometriosis but have not been diagnosed). Results . We identify a unique subcluster of proliferating uterine natural killer (uNK) cells in ME-tissues from controls that is almost absent from endometriosis cases, along with a striking reduction of total uNK cells in the ME of cases (p<10 -16 ). In addition, an IGFBP1+ decidualized subset of endometrial stromal cells are abundant in the shed endometrium of controls when compared to cases (p<10 -16 ) confirming findings of compromised decidualization of cultured stromal cells from cases. By contrast, endometrial stromal cells from cases are enriched in cells expressing pro-inflammatory and senescent phenotypes. An enrichment of B cells in the cases (p=5.8 x 10 -6 ) raises the possibility that some may have chronic endometritis, a disorder which predisposes to endometriosis. Conclusions . We propose that characterization of endometrial tissues in ME will provide an effective screening tool for identifying endometriosis in patients with chronic symptoms suggestive of this disorder. This constitutes a major advance, since delayed diagnosis for many years is a major clinical problem in the evaluation of these patients. Comprehensive analysis of ME is expected to lead to new diagnostic and therapeutic approaches to endometriosis and other associated reproductive disorders such as female infertility.
Project description:We focus on the characterization of gene expression profiles in circulating monocytes and peritoneal fluid Mononuclear phagocytes in patients with endometriosis
Project description:Microbiome dysregulation affects the estrogen metabolism (estrobolome) profile which in turns affects the immunological response. Estrogen hormone is an essential hormone that regulates the sexual activity in females as well as immune response in both sexes. Endometriosis is one of complicated disorder influnce the fertility of females due to escape of endometrial tissue into peritoneal cavity where the immunotoxicity will be undertaken. This study designed to investigate if there is any correlation between the microbiome dyregulation with the severity of endometriosis outcomes.
Project description:Transcriptome profiles were investigated in isolated endometrial stromal cells (ESCs) from eutopic and ectopic endometrium. The profiles were quite different between eutopic ESC and ectopic ESC, whereas no clear dfference was recognized between eutopic ESC with and without endometriosis. Total RNA from three cultured endometrial stromal cells (ESCs) from eutopic endometria without endometriosis, three ESCs with endometriosis and three ESCs from chocolate cysts were hybridised to the Affymetrix Human Gene 1.0 ST Array.
Project description:Profiles of genome-wide DNA methylation were investigated in isolated endometrial stromal cells from eutopic and ectopic endometrium. DNA methylation profiles were quite different between eutopic ESC and ectopic ESC, whereas no clear dfference were recognized between eutpic ESC with and without endometriosis. Bisulphite converted DNA from three cultured endometrial stromal cells (ESCs) from eutopic endometria without endometriosis, three ESCs with endometriosis and three ESCs from chocolate cysts were hybridised to the Illumina infinium HumanMethylation27 BeadChip.