Potential Biomarkers For Endometriosis Research Articles

Endometriotic lesions and their recurrence: A Study on the mediators of immunoregulatory (TGF-β/miR-20a) and stemness (NANOG/miR-145)

Endometriosis is a common estrogen-dependent disease that involves various cellular processes. Additionally, miRNAs play a crucial role in the development of the disease as an important component of the microenvironment. In this study, tissue specimens of eutopic and ectopic lesions of 20 women, whose endometriosis was later approved by the pathology laboratory, were biopsied through laparoscopy. As a control group, endometrial tissue specimens were collected from 20 women who underwent curettage for reasons unrelated to endometriosis. The expression levels of miR-20A and miR-145 and their target genes, TGF-β and NANOG, were measured in these samples as markers of stemness and immunomodulatory properties, respectively. The study also aimed to compare the expression levels of target genes and miRNAs in ectopic lesions regarding endometriosis recurrence post-surgery. The study revealed that the expression of TGF-β and NANOG genes was significantly upregulated in endometriotic tissues compared to the control group. There was also a notable increase in miR-20A and miR-145 expression in the endometriotic tissues compared to the control group. While there was no significant correlation between the expression of miR-20a and TGF-β, we observed a negative correlation between the expression level of miR-145 and NANOG. Additionally, the ROC curve analysis emphasized miR-14 as a potential biomarker for endometriosis over miR-20a. However, our findings on disease recurrence underscore the importance of miR-20a in the early detection of endometriosis recurrence.

PARP-1, EpCAM, and FRα as potential targets for intraoperative detection and delineation of endometriosis: a quantitative tissue expression analysis

BackgroundEndometriosis is a gynecological disease characterized by the presence of endometrial tissue in abnormal locations, leading to severe symptoms, inflammation, pain, organ dysfunction, and infertility. Surgical removal of endometriosis lesions is crucial for improving pain and fertility outcomes, with the goal of complete lesion removal. This study aimed to analyze the location and expression patterns of poly (ADP-ribose) polymerase 1 (PARP-1), epithelial cell adhesion molecule (EpCAM), and folate receptor alpha (FRα) in endometriosis lesions and evaluate their potential for targeted imaging.MethodsGene expression analysis was performed using the Turku endometriosis database (EndometDB). By immunohistochemistry, we investigated the presence and distribution of PARP-1, EpCAM, and FRα in endometriosis foci and adjacent tissue. We also applied an ad hoc platform for the analysis of images to perform a quantitative immunolocalization analysis. Double immunofluorescence analysis was carried out for PARP-1 and EpCAM, as well as for PARP-1 and FRα, to explore the expression of these combined markers within endometriosis foci and their potential simultaneous utilization in surgical treatment.ResultsGene expression analysis revealed that PARP-1, EpCAM, and FOLR1 (FRα gene) are more highly expressed in endometriotic lesions than in the peritoneum, which served as the control tissue. The results of the immunohistochemical study revealed a significant increase in the expression levels of all three biomarkers inside the endometriosis foci compared to the adjacent tissues. Additionally, the double immunofluorescence analysis consistently demonstrated the presence of PARP-1 in the nucleus and the expression of EpCAM and FRα in the cell membrane and cytoplasm.ConclusionOverall, these three markers demonstrate significant potential for effective imaging of endometriosis. In particular, the results emphasize the importance of PARP-1 expression as a possible indicator for distinguishing endometriotic lesions from adjacent tissue. PARP-1, as a potential biomarker for endometriosis, offers promising avenues for further investigation in terms of both pathophysiology and diagnostic-therapeutic approaches.

Determination of Serum Arginase-1 Concentrations and Serum Arginase Activity for the Non-Invasive Diagnosis of Endometriosis.

Backgroud: Endometriosis remains a diagnostic challenge, both clinically and economically, affecting 6% to 15% of women of child-bearing potential. We have attempted to determine whether testing serum concentrations and activity of arginase isoenzymes could be useful for the non-invasive diagnosis of endometriosis. Methods: This study involved 180 women (105 endometriosis subjects-study group B; 22 subjects with other benign gynaecological conditions-control group 1-K1, both undergoing surgery; and 53 healthy subjects without features of endometriosis-control group 2-K2). Results: Preoperative and postoperative arginase-1 (Arg-1) concentrations were significantly higher in patients, as compared with the control groups K1 (p < 0.0001 and p = 0.0005, respectively) and K2 (both p < 0.0001). Similarly, arginase activity was significantly higher in patients than in the control group K1 before surgery and higher than in both control groups after surgery. No significant differences in either Arg-1 concentrations or arginase activity were noted between the operated control group K1 and the non-operated control group K2. A significant postoperative decrease in Arg-1 concentration was observed within both patient (p < 0.0001) and control group K1 (p = 0.0043). Diagnostic performance was assessed using the receiver operating characteristic (ROC) method. The threshold for differentiation between endometriosis patients and healthy non-operated controls was 42.3 ng/mL, with a sensitivity of 90% and specificity of 81%. For differentiation of patients and operated controls with benign gynaecological conditions, the threshold was 78.4 ng/mL, with a sensitivity of 61% and specificity of 95%. Conclusions: We, therefore, conclude that Arg-1 serum concentrations and arginase activity could be considered potential biomarkers for endometriosis but require further studies on larger cohorts of patients.

ITRAQ-based proteomics analysis of human ectopic endometrial stromal cells treated by Maqian essential oil

BackgroundEndometriosis is a common and complex syndrome characterized by the presence of endometrial-like tissue outside the uterus. Chinese medicine has been recently found to show good efficacy in treating endometriosis. Our previous results revealed that Maqian fruit essential oil (MQEO) could inhibit the proliferation and induce apoptosis of ectopic endometrial stromal cells (EESCs), but the mechanisms remain unclear. In this study, we aim to explore the molecular mechanism of MQEO’s specific effects in EESCs.MethodsWe conducted a quantitative proteomics analysis by iTRAQ on EESCs treated with MQEO or DMSO. Then deep analysis was performed based on differentially expressed proteins, including Gene Ontology enrichment analysis, pathway enrichment analysis and protein interaction analysis. Candidate protein targets were subsequently verified by western blotting.ResultsAmong 6575 identified proteins, 435 proteins exhibited altered expression levels in MQEO-treated EESCs. Of these proteins, most were distributed in signal transduction as well as immune system and the most significantly altered pathway was complement and coagulation cascades. Moreover, two differentially expressed proteins (Heme oxygenase 1 and Acyl-CoA 6-desaturase) were verified and they can be potential biomarkers for endometriosis treatment.ConclusionsOur proteomic analysis revealed distinct protein expression patterns induced by MQEO treatment in EESCs, highlighting the potential of MQEO for endometriosis treatment and biomarker discovery.

Identification and validation of M2 macrophage-related genes in endometriosis

AimsM2 macrophage is believed to play an important role in the development of endometriosis. This study aimed to identify several key genes related to the M2 macrophage in endometriosis. MethodDifferential expressed genes between endometriosis and non-endometriosis were identified based on three microarray datasets from the Gene Expression Omnibus database. Gene modules significantly associated with M2 macrophage were identified from the weighted gene co-expression network analysis. Furthermore, by intersecting the differential expressed genes and M2 macrophage-associated module genes, M2 macrophage-related genes in endometriosis were identified. Functional analyses of the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes for these genes were then performed. Following, the least absolute shrinkage and selection operator, random forest, and receiver operating characteristic curves were further conducted to identify the key M2 macrophage-related genes in endometriosis. Finally, the expressions of key genes in endometriosis, as well as their correlations with M2 macrophages were verified in an independent validation cohort. ResultsTotally, 185 M2 macrophage-related genes were identified, and they were mainly enriched in functions associated with the cell cycle, oocyte maturation, and immune response. Following machine learning algorithms, eight key genes were selected in the endometriosis: PGR, OLFM4, PIP5K1B, CCNA1, BRIP1, CADM1, PRAME, and GCNT1. The eight key genes were confirmed to be negative with M2 macrophage infiltration levels. Furthermore, the expression levels of these genes were significantly lower in the middle secretory stage while relevantly higher in the proliferative stage. The validation analysis also showed similar outcomes with the above results. ConclusionEight M2 macrophage-related genes were identified as potential biomarkers of endometriosis, providing novel understanding of immune cells in the endometriosis.

MiRNA Expression Profiles in an Ectopic Endometrium of Patients at Different Stage of Endometriosis

Endometriosis is a debilitating disease that affects up to 15% of women worldwide. Delayed diagnosis, lack of noninvasive biomarker and unexplained pathogenesis are key problems for specialists and patients alike. MiRNAs are a class of noncoding RNAs that regulate myriad of cellular functions. These gene expression regulators may prove to be attractive biomarkers with diagnostic and prognostic value for endometriosis.&#x0D; Tissue samples were collected from the participants enrolled in the study during laparoscopy (patients) and hysteroscopy (control group). After RNA isolation (miRNeasy MiniKit, Qiagen) based on the disease stage, three pools – each containing 15 RNA samples, were constructed: 1) early stage endometriosis, 2) late stage endometriosis, and 3) healthy controls. Each pool sample was subjected to reverse transcription via miScript II RT Kit, Qiagen to obtain cDNA. SYBR Green based Real-time PCR assay was used to determine the expression profile of 84 miRNAs (Human miFinder miRNA PCR Array, Qiagen).&#x0D; We detected 32 differently expressed miRNAs between early stage endometriosis and control group, and 51 differently expressed miRNAs between advanced stage endometriosis and control group. Three miRNAs were differentially expressed with more than 10-fold change in the early stage group and thn miRNAs showed more than 10-fold change in the advanced stages group. The three miRNAs with the highest fold change in the expression levels in both case groups were nominated as potential biomarkers for endometriosis. The results of the analysis of their target genes supports the role of deregulation of apoptosis, angiogenesis, epithelial-mesenchymal transition, and various other cellular signalling pathways in endometriosis development.

FZD7: A potential biomarker for endometriosis.

Endometriosis is a chronic inflammatory, benign disorder that often co-occurs with adenomyosis and/or leiomyoma. The overall incidence of endometriosis in reproductive period women was nearly 10%. However, the exact mechanisms of endometriosis-associated pathogenesis are still unknown. In this study, we aimed to investigate whether Frizzled-7 (FZD7) would effectively promote the development of endometriosis. The microarray-based data analysis was performed to screen endometriosis-related differentially expressed genes. This process uncovered specific hub genes, and the nexus of vital genes and ferroptosis-related genes were pinpointed. Then, we collected human endometrial and endometriotic tissues from patients with endometriosis of the ovary (n = 39) and control patients without endometriosis (n = 10, who underwent hysterectomy for uterine fibroids) to compare the expression of FZD7. These findings indicated that the expression of FZD7 was high compared with normal endometrium, and FZD7 may promote the progression of endometriosis. FZD7 may serve as a potential therapeutic target for endometriosis treatment.

Transcriptomic analysis of granulosa cells in patients with endometriosis-related infertility: identification of potential molecular mechanisms

Objective: To investigate the impact of endometriosis on granulosa cell function and elucidate the molecular mechanisms involved. Methods: RNA sequencing, differential expression analysis, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, gene set enrichment analysis, protein-protein interaction analysis, and RT-qPCR were employed to assess the effects of endometriosis on granulosa cell function. Results: The results revealed significant differences in gene expression between the endometriosis and control groups, including genes related to immune regulatory functions and ferroptosis. Hub gene modules and hub genes were identified, including those related to cell cycle and immune and inflammatory pathways. RT-qPCR revealed significant up-regulation of the hub genes CCL3 and IL1B in granulosa cells of patients with endometriosis. Conclusion: The results of RNA sequencing demonstrated that endometriosis induces significant transcriptional alterations in granulosa cells of affected patients. These findings provide important insights into the diagnosis and treatment of endometriosis and highlight the importance of further investigation of CCL3 and IL1B as potential biomarkers for endometriosis.

Identification and Analysis of Potential Immune-Related Biomarkers in Endometriosis.

Endometriosis is an inflammatory gynecological disease leading to deep pelvic pain, dyspareunia, and infertility. The pathophysiology of endometriosis is complex and depends on a variety of biological processes and pathways. Therefore, there is an urgent need to identify reliable biomarkers for early detection and accurate diagnosis to predict clinical outcomes and aid in the early intervention of endometriosis. We screened transcription factor- (TF-) immune-related gene (IRG) regulatory networks as potential biomarkers to reveal new molecular subgroups for the early diagnosis of endometriosis. To explore potential therapeutic targets for endometriosis, the Gene Expression Omnibus (GEO), Immunology Database and Analysis Portal (ImmPort), and TF databases were used to obtain data related to the recognition of differentially expressed genes (DEGs), differentially expressed IRGs (DEIRGs), and differentially expressed TFs (DETFs). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed on the DETFs and DEIRGs. Then, DETFs and DEIRGs were further validated in the external datasets of GSE51981 and GSE1230103. Then, we used quantitative real-time polymerase chain reaction (qRT-PCR) to verify the hub genes. Simultaneously, the Pearson correlation analysis and protein-protein interaction (PPI) analyses were used to indicate the potential mechanisms of TF-IRGs at the molecular level and obtain hub IRGs. Finally, the receiver operating characteristic (ROC) curve analysis was used to assess the diagnostic value of the hub IRGs. We screened a total of 94 DETFs and 121 DEIRGs in endometriosis. Most downregulated DETFs showed decreased expression in the endometria of moderate/severe endometriosis patients. The top-ranked upregulated DEIRGs were upregulated in the endometra of infertile women. Functional analysis showed that DETFs and DEIRGs may be involved in the biological behaviors and pathways of endometriosis. The TF-IRG PPI network was successfully constructed. Compared with the control group, high C3, VCAM1, ITGB2, and C3AR1 expression had statistical significance in endometriosis among the hub DEIRGs. They also showed higher sensitivity and specificity by ROC analysis for the diagnosis of endometriosis. Finally, compared with controls, C3 and VCAM1 were highly expressed in endometriosis tissue samples. In addition, they also showed high specificity and sensitivity for diagnosing endometriosis. Overall, we discovered the TF-IRG regulatory network and analyzed 4 hub IRGs that were closely related to endometriosis, which contributes to the diagnosis of endometriosis. Additionally, we verified that DETFs or DEIRGs were associated with the clinicopathological features of endometriosis, and external datasets also confirmed the hub IRGs. Finally, C3 and VCAM1 were highly expressed in endometriosis tissue samples compared with controls and may be potential biomarkers of endometriosis, which are helpful for the early diagnosis of endometriosis.

Exosomal miR-202 derived from leukorrhea as a potential biomarker for endometriosis

Endometriosis (EMS) is a chronic gynecological disorder with an urgent need of a reliable non-invasive diagnostic strategy. Recently, there has been increasing interest in using the contents of exosomes, especially exosomal microRNAs (miRNAs), as potential biomarkers for various types of diseases. In this study, we assessed the differentially expressed miRNAs in exosomes derived from primary normal and ectopic endometrial cells. We used miRNA microarray analysis to identify differentially expressed exosomal miRNAs. Among the selected exosomal miRNAs, we focused on hsa-miR-202-3p and hsa-miR-202-5p and validated their expression levels using quantitative reverse transcription polymerase chain reaction analysis. We then further examined their expression in exosomes derived from vaginal discharge (leukorrhea) from patients with EMS and the negative control group. The data show that hsa-miR-202-3p and hsa-miR-202-5p were expressed significantly higher in leukorrhea-derived exosomes from EMS patients compared with the negative controls. Taken together, our results suggest that leukorrhea-derived exosomal hsa-miR-202 could serve as a potential useful biomarker for diagnosing EMS.

Identification and Validation of Three m6A Regulators: FTO, HNRNPC, and HNRNPA2B1 as Potential Biomarkers for Endometriosis.

N6-methyladenosine is involved in numerous biological processes. However, the significance of m6A regulators in endometriosis is still unclear. We extracted three significant m6A regulators between non-endometriosis and endometriosis patients from GSE6364 and then we used the random forest model to obtain significant m6A regulators. In addition, we used the nomogram model to evaluate the prevalence of endometriosis. The predictive ability of the candidate genes was evaluated through the receiver operating characteristic curves, while the expression of candidate biomarkers was validated via Western blotting. Additionally, according to candidate genes, we identified m6A subtypes based on which functional enrichment analysis and immune infiltration were performed. Three significant m6A regulators (fat mass and obesity-associated protein, heterogeneous nuclear ribonucleoprotein A2/B1, and heterogeneous nuclear ribonucleoprotein C) were discovered. We identified three m6A subtypes, including clusterA, clusterB, and clusterC. ClusterB was demonstrated to be correlated with significantly overexpressed VEGF and notably downregulated ESR1 and PGR, which are convincing biomarkers of endometriosis. Furthermore, we discovered that patients in clusterB were associated with high levels of neutrophil infiltration, a reduced Treg/Th17 ratio, and overexpressed pyroptosis-related genes, which also indicated that clusterB was highly linked to endometriosis. In conclusion, m6A regulators are of great significance for the occurrence and process of endometriosis. The findings of our study provide novel insights into the underlying molecular mechanism of endometriosis. The novel investigation of m6A patterns and their correlation with immunity may also help to guide the clinical diagnosis, provide prognostic significance, and develop immunotherapy strategies for endometriosis patients.

Plasma and Peritoneal Fluid Fibronectin and Collagen IV Levels as Potential Biomarkers of Endometriosis.

Laparoscopy as a diagnostic tool for patients with suspected endometriosis is associated with several potentially life-threatening complications. Therefore, it is imperative to identify reliable, non-invasive biomarkers of the disease. The aim of this study was to analyse the concentrations of fibronectin and type IV collagen in peritoneal fluid and plasma to assess their role as potential biomarkers in the diagnosis of endometriosis. Fibronectin and collagen IV protein levels were assessed by surface plasmon resonance imaging (SPRi) biosensors with the usage of monoclonal antibodies. All patients enrolled in the study were referred for laparoscopy for the diagnosis of infertility or chronic pelvic pain (n = 84). The study group included patients with endometriosis confirmed during surgery (n = 49). The concentration of fibronectin in the plasma (329.3 ± 98.5 mg/L) and peritoneal fluid (26.8 ± 11.1 μg/L) in women with endometriosis was significantly higher than in the control group (251.2 ± 84.0 mg/L, 7.0 ± 5.9 μg/L). Fibronectin levels were independent of endometriosis stage (p = 0.874, p = 0.469). No significant differences were observed in collagen IV levels (p = 0.385, p = 0.465). The presence of elevated levels of fibronectin may indicate abnormalities in cell-ECM signalling during the course of endometriosis, and may be a potential biomarker for early detection.

Gene expression analysis in endometriosis: Immunopathology insights, transcription factors and therapeutic targets.

Endometriosis is recognized as an estrogen-dependent inflammation disorder, estimated to affect 8%-15% of women of childbearing age. Currently, the etiology and pathogenesis of endometriosis are not completely clear. Underlying mechanism for endometriosis is still under debate and needs further exploration. The involvement of transcription factors and immune mediations may be involved in the pathophysiological process of endometriosis, but the specific mechanism remains to be explored. This study aims to investigate the underlying molecular mechanisms in endometriosis. The gene expression profile of endometriosis was obtained from the gene expression omnibus (GEO) database. Gene set variation analysis (GSVA) and gene set enrichment analysis (GSEA) were applied to the endometriosis GSE7305 datasets. Cibersort and MCP-counter were used to explore the immune response gene sets, immune response pathway, and immune environment. Differentially expressed genes (DEGs) were identified and screened. Common biological pathways were being investigated using the kyotoencyclopediaofgenesandgenomes (KEGG) pathway enrichment analysis. Transcription factors were from The Human Transcription Factors. The least absolute shrinkage and selection operator (Lasso) model identified four differential expressions of transcription factors (AEBP1, HOXB6, KLF2, and RORB). Their diagnostic value was calculated by receiver operating characteristic (ROC) curve analysis and validated in the validation cohort (GSE11691, GSE23339). By constructing the interaction network of crucial transcription factors, weighted gene coexpression network analysis (WGCNA) was used to search for key module genes. Metascape was used for enrichment analysis of essential module genes and obtained HOXB6, KLF2. The HOXB6 and KLF2 were further verified as the only two intersection genes according to Support Vector Machine Recursive Feature Elimination (SVM-RFE) and random forest models. We constructed ceRNA (lncRNA-miRNA-mRNA) networks with four potential transcription factors. Finally, we performed molecular docking for goserelin and dienogest with four transcription factors (AEBP1, HOXB6, KLF2, and RORB) to screen potential drug targets. Immune and metabolic pathways were enriched in GSVA and GSEA. In single sample gene set enrichment analysis (ssGSEA), most immune infiltrating cells, immune response gene sets, and immune response pathways are differentially expressed between endometriosis and non-endometriosis. Twenty-seven transcription factors were screened from differentially expressed genes. Most of the twenty-seven transcription factors were correlated with immune infiltrating cells, immune response gene sets and immune response pathways. Furthermore, Adipocyte enhancer binding protein 1 (AEBP1), Homeobox B6 (HOXB6), Kruppel Like Factor 2 (KLF2) and RAR Related Orphan Receptor B (RORB) were selected out from twenty-seven transcription factors. ROC analysis showed that the four genes had a high diagnostic value for endometriosis. In addition, KLF2 and HOXB6 were found to play particularly important roles in multiple modules (String, WGCNA, SVM-RFE, random forest) on the gene interaction network. Using the ceRNA network, we found that NEAT1 may regulate the expressions of AEBP1, HOXB6 and RORB, while X Inactive Specific Transcript (XIST) may control the expressions of HOXB6, RORB and KLF2. Finally, we found that goserelin and dienogest may be potential drugs to regulate AEBP1, HOXB6, KLF2 and RORB through molecular docking. AEBP1, HOXB6, KLF2, and RORB may be potential biomarkers for endometriosis. Two of them, KLF2 and HOXB6, are critical molecules in the gene interaction network of endometriosis. Discovered by molecular docking, AEBP1, HOXB6, KLF2, and RORB are targets for goserelin and dienogest.

O-027 Small extracellular vesicle (sEV) protein cargo as potential biomarker for endometriosis

Abstract Study question Can an endometriosis-specific protein signature in small extracellular vesicles (sEV) from peritoneal fluid (PF) be utilised as a non-invasive biomarker of the condition? Summary answer Yes, potentially. We found differences in the concentrations and protein cargo of PF-derived sEV between controls and endometriosis samples, most notably in CD44 expression. What is known already Endometriosis, defined as endometrial-like tissue outside the uterus, causes pain and/or subfertility in 10% of reproductive age women. The cause is unknown, resulting in inadequate diagnostic methods and treatment options. There is no clinically relevant biomarker for endometriosis yet. Small extracellular vesicles (sEV), produced by virtually every cell, have been described in diseases such as cancer, diabetes, and pre-eclampsia, and could similarly be important in endometriosis. We previously identified sEV in PF of women with endometriosis, and here investigated the protein cargo of PF sEV as biomarker of the disease. Study design, size, duration PF samples were obtained from participants in the ENDOX study, Endometriosis CaRe Centre, Nuffield Department of Women’s and Reproductive Health, University of Oxford (REC ref. 09/H0604/58) according to WERF EPHect standards. Women between 18-49 years of age (n = 63) who had undergone diagnostic laparoscopy were classified according to cycle phase (proliferative/secretory/menstrual) and severity of endometriosis (ASRM stages I+II or stages III+IV). Exclusion criteria were hormonal treatment, malignancy, pregnancy, breastfeeding, and inability to understand the consent form. Participants/materials, setting, methods The participant groups were control proliferative, n = 7; control secretory, n = 9; control menstrual, n = 3; StI+II proliferative, n = 8; StI+II secretory, n = 10; St1+II menstrual, n = 7; StIII+IV proliferative, n = 5; StIII+IV secretory, n = 11; StIII+IV menstrual, n = 3. 1 mL PF was centrifuged to remove cells, debris, and microvesicles. sEV were isolated using size exclusion chromatography (SEC) and analysed by nanoparticle tracking analysis (NTA), immunoblotting, and mass spectrometry (LC-MS/MS). Main results and the role of chance We confirmed the presence of exosomes in PF from women at different stages of endometriosis and from disease-free patients at different menstrual cycle phases by NTA, immunoblotting and mass spectrometry. Enriched sEV were positive for ALIX, CD9, and syntenin. The mode size of PF particles from women with endometriosis was 115 ± 15.5 nm, whereas in non-endometriotic women it was 95 ± 17.3 nm (n.s.). sEV concentrations were higher in endometriosis compared to controls, and highest in stage III-IV endometriosis, followed by stage I-II endometriosis and controls, irrespective of menstrual cycle phase (P = 0.0210). sEV concentration in stage III-IV endometriosis decreased consistent with a transition from proliferative to secretory phase. Likewise, PF-derived sEV numbers within stage I-II endometriosis samples increased, as these samples transitioned from proliferative to secretory cycle phases. Proteomic analysis showed distinct distribution patterns of proteins within endometriosis PF-derived sEVs compared to controls. Consistent with earlier studies, we found CD44 as an sEV protein uniquely within the endometriosis population and contributing significantly to the separation of endometriosis and control samples by the highest variable importance projection (VIP) score in our data set. Limitations, reasons for caution The main limitation of this study is the small number of samples across the different groups, and the limited amount of PF per sample. Wider implications of the findings PF-derived sEV differ between endometriosis and control patients. Concentrations vary regardless of cycle phase and disease stage, and this difference appears to be reflected in the proteomics analysis. The presence of CD44 within sEV could help diagnose endometriosis. Trial registration number not applicable

Evaluation of Soluble CD90: Potential for Diagnostic Significance in Endometriosis Patients.

Background Endometriosis is a chronic and debilitating gynecologic disorder, driven by endocrine and immune dysfunctions, which lead to poor endometrial differentiation and attenuated fertility. Escape from immune surveillance and involvement of inflammatory mechanisms appear to be factors in disease progression. Current diagnostic guidelines for endometriosis still lack an efficient biomarker. Here, we report a study on two previously unexplored factors as potential biomarkers for endometriosis. Methods A case-control study was performed to evaluate the diagnostic potential of serum CD90 and CD83 levels in endometriosis patients (cases validated by surgical and histological examination) compared to healthy controls. Serum was collected from age-matched females and analyzed by ELISA. Results Comparison of endometriosis patients to the control group showed significantly elevated levels of serum CD90 (1160 ± 856 pg/mL vs. 334 ± 228 pg/mL; ∗∗∗∗p < 0.0001). A threshold value of 479.4 pg/mL was defined based on the control results, and the diagnostic efficiency of the test was estimated. The obtained sensitivity (70.4%), specificity (92.9%), positive predictive value (90.5%), and negative predictive value (76.5%) rated the test as one with promising diagnostic potential. In contrast, the analysis of serum CD83 levels showed comparable values in both groups, suggesting no association with patient status. Conclusion Elevated soluble CD90 in human serum is associated with endometriosis, which suggests its putative clinical significance as a biomarker in screening and/or diagnosis of the disease.

Non-invasive diagnostics of endometriosis: review of modern biomarkers of peripheral blood and endometrium

Endometriosis is a chronic progressing estrogen-dependent disease with a high incidence among females characterized by pelvic pain (40-80%) and infertility (25-80%).&#x0D; Aim. To analyze and estimate the modern literature data on the possibility of using potential biomarkers of endometriosis in its non-invasive diagnostics.&#x0D; In recent years, researchers have made significant advances in understanding the disease-specific molecular pathways that regulate the development of ectopic foci of endometriosis, by examining the blood, peritoneal fluid, and eutopic endometrium in women with the disease. Along with understanding the pathophysiology of endometriosis, the question of finding an adequate biomarker that will provide an effective early non-invasive diagnosis of endometriosis and, accordingly, preserve the reproductive health of millions of women, remains relevant.&#x0D; Conclusion. Most reasonable in diagnostics of endometriosis is a combination of different biomarkers that also minimizes false positive and negative results in differential diagnosis.

Expression of nerve growth factor (NGF) in endometrium as a potential biomarker for endometriosis – Single tertiary care centre study

Objectiveto identify novel biomarkers for peritoneal endometriosis in eutopic endometrium thus giving an oportunity for non-invasive diagnosis. DesignA cross-sectional single-center study Settingtertiary care hospital Patients49 patients subjected to laparoscopy because of suspected endometriosis, 33 patients out of the group qualified to the study had sufficient endometrial tissue taken and were in their follicular phase of menstrual cycle. Interventionsbiopsy sampling of eutopic endometrial tissue during diagnostic or diagnostic and terapeutic laparoscopy, questionaires, Main Outcome Measure(s)qRT-PCR to evaluate the mRNA expression of selected candidate marker genes in endometrium: ARO1 (aromatase), CXCL8 (interleukin 8), NGF (nerve growth factor), VEGF-A (vascular endothelial growth factor A), PDGF-A (platelet-derived growth factor A). ResultsmRNA expression of ARO1, CXCL8, VEGF-A and PDGF-A did not differ significantly between women with and without endometriosis. NGF mRNA expression was decreased in women with endometriosis. ConclusionsObserved preliminary results suggest a possible role of NGF in early diagnosis of peritoneal endometriosis. The role of NGF changes in eutopic endometrium of patients with peritoneal endometriosis needs further evaluation.

DETERMINATION OF ANTIGEN LEVELS: T-PLASTIN, TRANSGELIN AND CA-125 IN BABOONS WITH INDUCED ENDOMETRIOSIS

Endometriosis is defined as the presence of endometrial-like tissue outside the uterine cavity and is associated with chronic intra-pelvic inflammation. Its symptoms, which are often severe, have a negative impact on a woman’s quality of life. They include chronic pelvic pain and infertility. It is estimated that about 10% women in their reproductive ages are affected by the disease globally. Currently, diagnostic methods are laparoscopy with histological confirmation and non-invasive such as ultrasound and the quantification of CA-125 biomarker in serum. CA-125 is known to be elevated in endometriosis, however it does not have sufficient diagnostic power as a single biomarker of endometriosis. Due to this, there is a need to determine other antigens that would act as potential biomarkers for endometriosis. It has been observed that Transgelin and T-Plastin are upregulated in endometriosis lesions and in secretory phase of endometrium respectively. There is insufficient data available showing the concentration of T-plastin in serum or plasma in the development of endometriosis. The main objective of this study was to identify whether T-plastin, Transgelin and CA-125 are potential biomarkers in early diagnosis of endometriosis using animal model for endometriosis. The baboon model represents clinically relevant research models for endometriosis. This study compared the levels of T-plastin, Transgelin and CA-125 in the peripheral blood and peritoneal fluid of baboons before and after induction of endometriosis. In this prospective, case-control study, ten female baboons ( Papio anubis , 9-15 kg each) of proven fertility that have had at least one menstruation during captivity, were induced by intra-pelvic injection of menstrual endometrium on day 1 or 2 of menstruation followed by staging laparoscopy. Serum and peritoneal fluid samples were collected prior to induction to serve as controls.T-Plastin, Transgelin and CA-125 were measured using commercially available ELISA Kits. Data was analyzed using non-parametric test and level of significance at p&lt;0.005. T-Plastin and Transgelin had insignificant p values in both peripheral blood and peritoneal fluid while CA-125 showed p=0.0003 in peripheral blood and p=0.0279 in peritoneal fluid. Diagnostic performance of individual biomarkers was determined by Receiver Operating Characteristic (ROC) with the Area under the curve (AUC) equal to 0.5 showing T-Plastin and Transgelin in both serum and peritoneal fluid unreliable for use as biomarkers in endometriosis. CA-125 levels in peritoneal fluid had an AUC of 0.7900 indicating it can be used as potential biomarkers for endometriosis. Key words: Endometriosis, biomarkers, T-Plastin, Transgelin, CA-125, ELISA, Olive Baboons. DOI : 10.7176/ALST/75-06 Publication date :August 31 st 2019

Elevated serum interleukin-32 levels in patients with endometriosis: A cross-sectional study.

Recently, interleukin (IL)-32 has been suggested to be involved in the pathogenesis of endometriosis. The aim of this study was to investigate whether serum IL-32 level might be used as a biomarker for diagnosis of endometriosis. We recruited the serum samples of 50 patients with histologically confirmed endometriosis and 35 controls. Enzyme-linked immunosorbent assay was used to analyze the serum IL-32, IL-6, IL-10, tumour necrosis factor (TNF)-α, IL-1β, and CA-125 levels in patients with and without the disease and the diagnostic potentials of the cytokines were assessed using receiver operating characteristic curve and the area under the curve (AUC). Among evaluated cytokines, only serum IL-32 levels showed significant differences between patients with and without endometriosis (1111.24±149.59 vs 631.10±120.23ng/mL, P=0.018, respectively). When the diagnostic power of serum IL-32 was evaluated, the AUC was 0.638 (95% confidence interval (CI): 0.521-0.766, P=0.031). When serum IL-32 levels were combined with serum CA-125 levels, the AUC was increased to 0.749 (95% CI: 0.640-0.858, P<0.001) with sensitivity and specificity of 60.0% and 82.9% at cutoff value of 0.640, which led to detect 25 more cases of endometriosis than the use of serum CA 125 with the cutoff value of 35IU/mL (36/50 vs 11/50, P<0.001) without sacrificing the specificity of the marker. Serum IL-32 levels are elevated in patients with endometriosis, and with combination of serum CA-125 levels, it may serve as a potential biomarker for endometriosis.

Identification of potential endometriosis biomarkers in peritoneal fluid and blood plasma via shotgun lipidomics

Endometriosis is a recurrent and benign gynecological disorder, defined by the ectopic presence of endometrium. About 10% of reproductive-aged women suffer from endometriosis. There are no non-invasive or minimally invasive tests available in clinical practice to accurately diagnose endometriosis today.Here, we present our efforts to determine the diagnostic accuracy of biomarkers in peritoneal fluid and blood plasma using flow injection analysis with electrospray ionization tandem mass spectrometry (ESI-MS/MS) in 70 women with endometriosis and 20 women from a control group. The presence of endometriosis was confirmed by surgical findings and post-operative pathological examination. A qualitative and quantitative evaluation of the lipids in peritoneal fluids and blood plasma was carried out using electrospray ionization mass spectrometry (ESI-MS). The analysis revealed more than 140 molecular species of lipids, most of which pertained to five classes: phosphatidylcholines, phosphatidylethanolamines, sphingomyelins, di- and triglycerides. The data were analyzed using a statistical multifactorial method (i.e., PLS-DA). It was found that 9 potential biomarkers of endometriosis (LPC 16:0, PE O-20:0, PE O 34:1, PC 36:2, PC 36:4, PC 36:5, PC 38:4, PC 38:6 and SM 34:1) are common in blood plasma and peritoneal fluid, supporting connection with the pathological process. The sensitivity of the method developed for plasma was 93% with a specificity of 95%; for peritoneal fluid, the sensitivity was 90% and the specificity 95%. Accordingly, plasma is the most suitable biological fluid for clinical diagnostics of endometriosis. Further validation of these lipids as serologic biomarkers may enhance non-invasive diagnostic tools for patients with suspected endometriosis and reduce the frequency of diagnostic laparoscopy.