Challenges in uncovering non-invasive biomarkers of endometriosis.
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ABSTRACT:
SUBMITTER: Hudson QJ
PROVIDER: S-EPMC7082884 | biostudies-literature | 2020 Mar
REPOSITORIES: biostudies-literature
Publications
Challenges in uncovering non-invasive biomarkers of endometriosis.
Hudson Quanah J QJ Perricos Alexandra A Wenzl Rene R Yotova Iveta I
Experimental biology and medicine (Maywood, N.J.) 20200204 5
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Project description:BackgroundEndometriosis is a common gynaecological disorder affecting 5-10% of women of reproductive age who often experience chronic pelvic pain and infertility. Definitive diagnosis is through laparoscopy, exposing patients to potentially serious complications, and is often delayed. Non-invasive biomarkers are urgently required to accelerate diagnosis and for triaging potential patients for surgery.MethodsThis retrospective case control biomarker discovery and validation study used quantitative 2D-difference gel electrophoresis and tandem mass tagging-liquid chromatography-tandem mass spectrometry for protein expression profiling of eutopic and ectopic endometrial tissue samples collected from 28 cases of endometriosis and 18 control patients undergoing surgery for investigation of chronic pelvic pain without endometriosis or prophylactic surgery. Samples were further sub-grouped by menstrual cycle phase. Selected differentially expressed candidate markers (LUM, CPM, TNC, TPM2 and PAEP) were verified by ELISA in a set of 87 serum samples collected from the same and additional women. Previously reported biomarkers (CA125, sICAM1, FST, VEGF, MCP1, MIF and IL1R2) were also validated and diagnostic performance of markers and combinations established.ResultsCycle phase and endometriosis-associated proteomic changes were identified in eutopic tissue from over 1400 identified gene products, yielding potential biomarker candidates. Bioinformatics analysis revealed enrichment of adhesion/extracellular matrix proteins and progesterone signalling. The best single marker for discriminating endometriosis from controls remained CA125 (AUC = 0.63), with the best cross-validated multimarker models improving the AUC to 0.71-0.81, depending upon menstrual cycle phase and control group.ConclusionsWe have identified menstrual cycle- and endometriosis-associated protein changes linked to various cellular processes that are potential biomarkers and that provide insight into the biology of endometriosis. Our data indicate that the markers tested, whilst not useful alone, have improved diagnostic accuracy when used in combination and demonstrate menstrual cycle specificity. Tissue heterogeneity and blood contamination is likely to have hindered biomarker discovery, whilst a small sample size precludes accurate determination of performance by cycle phase. Independent validation of these biomarker panels in a larger cohort is however warranted, and if successful, they may have clinical utility in triaging patients for surgery.
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Project description:Background/objectivesEndometriosis (END) is a painful gynecological condition. Clinical examination, imaging, and laparoscopy can provide a definitive diagnosis of END. Nonetheless, non-invasive biomarkers could help enhance and streamline the diagnostic process. Micro-RNAs (miRNAs), a family of small non-coding RNAs, could serve as useful non-invasive biomarkers for END. The aim of this study was to perform serum miRNA profiling in a retrospective cohort of women to identify miRNAs that are differentially expressed in END compared to control patients.MethodsRNA was isolated from serum samples of 67 END patients and 60 control women. The expression profile of a 754-miRNA panel was studied with RT-qPCR performed on a QuantStudio 12K Flex with the TaqMan OpenArray miRNA panel. A Censored Regression Model was used for miRNA differential expression analysis. Several gene-enrichment algorithms were employed to identify pathways related to the target genes of differentially expressed miRNAs.ResultsOne hundred and thirty miRNAs were detected in at least 75% of samples from either the END or the control group. Sixteen miRNAs were significantly modulated between the END and control groups. Enrichment analysis identified targets significantly overrepresented in numerous pathways involved in biological processes related to END, including inflammation, angiogenesis, cellular invasion, cell-cycle/cell proliferation, and estrogen and progesterone hormonal signaling.ConclusionsOur study indicates that differentially expressed miRNAs between END patients and controls can be identified through liquid biopsy. Our findings also suggest a potential role for serum miRNAs in the pathophysiology of END, warranting further investigations for their use as non-invasive biomarkers.
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Project description:CIRCULATING SERUM MICRO-RNA AS NON-INVASIVE DIAGNOSTIC BIOMARKERS OF ENDOMETRIOSIS
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Project description:Gastrointestinal mucositis is a complex inflammatory reaction of the mucous membranes, a side effect of both chemotherapy and radiotherapy. Currently, assessment scales are used to diagnose mucositis. However, a biomarker which would determine whether there is mucositis and thereby establish the severity objectively would be very useful. This will give the opportunity to evaluate studies, to determine risk factors and incidence, and it will make it possible to compare studies. Moreover, this biomarker might improve clinical management for patients. In this paper, we reviewed studies concerning potential biomarkers in blood samples and fecal samples, and potential tests in breath samples and urine samples. We include biomarkers and tests studied in animal models and/or in clinical trials, and discuss the validity, diagnostic accuracy, and applicability.
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Project description:Introduction: Endometriosis is a debilitating condition where endometrial-like tissue grows outside the uterus and rarely in distant sites such as the lungs, heart and brain. Symptoms are severe chronic pain, bladder/bowel problems, painful sex and infertility. Differences in endometrium between women with and without endometriosis have not been constructive, and there is still paucity in its pathogenesis and non-invasive diagnostic test, leading to delayed diagnosis (~10 years) and effective treatment. Menstrual fluid (MF) gives a peak into endometrial health. Could MF extracellular vesicles (EVs) point to the pathogenesis and a non-invasive diagnostic tool for endometriosis? Aims: To identify differences in protein-cargo in MF-EVs from women with and without endometriosis, define their functional contribution to mesothelial niche cells for lesion establishment and determine whether MF-sEV can be a non-invasive endometriosis diagnostic tool. Methods: MF was collected from women with and without endometriosis on day 2 of menses and EVs isolated using differential centrifugation. MF-EVs were identified by size, morphology and protein markers and subjected to TMT- quantitative proteomics assay. Differentially expressed proteins were analysed. Ctrl and Endo MF-sEV were co-cultured with mesothelial cells to assess for uptake and changes in the adhesive and junctional proteins. Results: Spherical-shaped MF-EVs were identified with mean diameters of 121.9 nm (Endo) and 123.8 nm (Ctrl), expressing TSG101, Alix and Syntenin-1 sEV proteins. MF-sEV proteins originated from endometrial cells: stem cells, leucocytes, and endothelial and smooth muscle cells, validating the endometrial origin of the MF-EVs. 77% of identified proteins were differentially expressed. In endo-MF-sEV, proteins regulating HGF receptor (anoikis), cellular senescence and multiple apoptotic, nitrogen compound metabolic processes and TRP channels signalling (immunity and contraction regulation) were decreased, potentially promoting lesion establishment, survival and pain. Similarly, antibacterial peptides (cathelicidin, mucin-1, lipocalin2 and serum amyloid A) were downregulated. The majority of proteins were significantly downregulated in endometriosis suggesting a dysregulation in the protein synthesis and cellular communication for homeostasis. Two-thirds of upregulated proteins were immunoglobulins indicating inflammatory pathogenesis. This was reflected by an increased Endo-MF-EVs uptake by mesothelial cells, with decreased scribble resulting in decreased cellular resistance and permeability. Conclusions: MF-EVs contain vital information indicative of endometriosis pathogenesis and may be the key to developing a simple and early diagnostic tool.
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