Multi-omics integration highlights the role of ubiquitination in endometriosis fibrosis.

BACKGROUND: Endometriosis, characterized by the presence of active endometrial-like tissues outside the uterus, causes symptoms like dysmenorrhea and infertility due to the fibrosis of endometrial cells, which involves excessive deposition of extracellular matrix (ECM) proteins. Ubiquitination, an important post-transcriptional modification, regulates various biological processes in human diseases. However, its role in the fibrosis process in endometriosis remains unclear. METHODS: We employed multi-omics approaches on two cohorts of endometriosis patients with 39 samples. GO terms and KEGG pathways enrichment analyses were used to investigate the functional changes involved in endometriosis. Pearson's correlation coefficient analysis was conducted to explore the relationship between global proteome and ubiquitylome in endometriosis. The protein expression levels of ubiquitin-, fibrosis-related proteins, and E3 ubiquitin-protein ligase TRIM33 were validated via Western blot. Transfecting human endometrial stroma cells (hESCs) with TRIM33 small interfering RNA (siRNA) in vitro to explore how TRIM33 affects fibrosis-related proteins. RESULTS: Integration of proteomics and transcriptomics showed genes with concurrent change of both mRNA and protein level which involved in ECM production in ectopic endometria. Ubiquitylomics distinguished 1647 and 1698 ubiquitinated lysine sites in the ectopic (EC) group compared to the normal (NC) and eutopic (EU) groups, respectively. Further multi-omics integration highlighted the essential role of ubiquitination in key fibrosis regulators in endometriosis. Correlation analysis between proteome and ubiquitylome showed correlation coefficients of 0.32 and 0.36 for ubiquitinated fibrosis proteins in EC/NC and EC/EU groups, respectively, indicating positive regulation of fibrosis-related protein expression by ubiquitination in ectopic lesions. We identified ubiquitination in 41 pivotal proteins within the fibrosis-related pathway of endometriosis. Finally, the elevated expression of TGFBR1/α-SMA/FAP/FN1/Collagen1 proteins in EC tissues were validated across independent samples. More importantly, we demonstrated that both the mRNA and protein levels of TRIM33 were reduced in endometriotic tissues. Knockdown of TRIM33 promoted TGFBR1/p-SMAD2/α-SMA/FN1 protein expressions in hESCs but did not significantly affect Collagen1/FAP levels, suggesting its inhibitory effect on fibrosis in vitro. CONCLUSIONS: This study, employing multi-omics approaches, provides novel insights into endometriosis ubiquitination profiles and reveals aberrant expression of the E3 ubiquitin ligase TRIM33 in endometriotic tissues, emphasizing their critical involvement in fibrosis pathogenesis and potential therapeutic targets.

Tamoxifen Alleviates Endometrial Fibrosis Induced by Anhydrous Ethanol in Rats.

BACKGROUND: As an estrogen receptor modulator, tamoxifen has been utilized in the treatment of fibrotic diseases. However, there is a limited body of research focusing on its potential application in addressing endometrial fibrosis conditions. Our research aims to investigate the effects of tamoxifen at different dosage levels in alleviating endometrial fibrosis and to elucidate its mechanisms of action during the initial stages of endometrial damage. METHODS: A total of thirty sexually mature, unmated female Sprague-Dawley (SD) rats were divided into six distinct groups. To establish the rat uterine adhesion model, the uterine cavity was subjected to perfusion with anhydrous ethanol. The control group received a saline solution, while the treatment group was administered oral estrogen in combination with tamoxifen at doses of 4 mg/kg/d, 20 mg/kg/d, and 40 mg/kg/d. Various techniques, including Hematoxylin and Eosin (HE) staining, Masson's Trichrome staining, Western Blotting analysis, and immunohistochemistry, were employed to assess changes in endometrial thickness, fibrosis, as well as alterations in indicators related to epithelial-mesenchymal transition (EMT), fibrosis, estrogen receptors within the endometrium, and vascular endothelial growth factor (VEGF). RESULTS: In the model group, the levels of endometrial thickness, E-cadherin, Vimentin, estrogen receptor α (ERα), G protein-coupled receptor 30 (GPR30), and VEGF proteins were significantly lower compared to the control group. Conversely, the levels of collagen accumulation, Smooth Muscle Actin (SMA), Transforming Growth Factor ß1 (TGFß1), Drosophila mothers against decapentaplegic protein 2 (Smad2) , and Smad3 were markedly higher than those observed in the control group (p < 0.05, p < 0.01, p < 0.001, p < 0.0001). In contrast, the low-dose tamoxifen group demonstrated significant increases in endometrial thickness, E-cadherin, Vimentin, ERα, GPR30, and VEGF when compared to the model group (p < 0.05, p < 0.01, p < 0.001, p < 0.0001). Moreover, the levels of collagen accumulation, TGFß1, SMA, Smad2, and Smad3, which are indicative of fibrosis and the TGFß1/Drosophila mothers against decapentaplegic (smad) pathway, were notably reduced compared to the model group (p < 0.05, p < 0.0001). CONCLUSIONS: The results of this study suggest that the administration of a low dose (4 mg/kg/d) of tamoxifen in the early stages of endometrial injury may mitigate epithelial-mesenchymal transition (EMT) indicators and reduce fibrosis within the endometrium induced by anhydrous ethanol.

Association Analysis between Maternal Serum Biomarkers and Fetal Congenital Heart Disease.

OBJECTIVE: To investigate the association of maternal serum biomarkers alpha fetal protein (AFP), ß-human chorionic gonadotropin (HCG) and unconjugated estriol 3 (uE3) in the second trimester with fetal congenital heart disease (CHD) in low risk populations during the screening of Down's syndrome. METHODS: 109 cases diagnosed with fetal CHD in the second trimester by fetal echocardiography were enrolled as the CHD group. Pregnancy- and gestational-age matched 344 cases without fetal CHD were used as the control group. The values of maternal serum biomarkers HCG, AFP and uE3 were tested and the association with CHD was analyzed. RESULTS: The means of HCG multiple of median (MoM) and AFP MoM were higher, while the mean of uE3 MoM was lower in the CHD group than those of the control group (p < 0.05). The number of cases with HCG MoM ≥85% quantile, AFP MoM ≥85% quantile was more, while that with uE3 MoM ≤15% quantile was less in the CHD group than that of the control group (p < 0.05). The univariate logistic regression analysis showed that fetal CHD was associated with high values of HCG MoM and AFP MoM and low value of uE3 MoM as well as the HCG MoM ≥85% quantile, AFP MoM ≥85% quantile and the uE3 MoM ≤15% quantile. The multivariate logistic regression analysis showed that HCG MoM ≥85% quantile and AFP MoM ≥85% quantile or the uE3 ≤15% quantile were the independent factors of fetal CHD. In addition, the risk of fetal CHD was higher when one situation existed among the HCG MoM ≥85% quantile, AFP MoM ≥85% quantile and the uE3 MoM ≤15% quantile. The risk of fetal CHD was much higher when both the HCG MoM ≥85% quantile and AFP MoM ≥85% quantile existed or both the HCG MoM ≥85% quantile and the uE3 MoM existed ≤15% quantile. CONCLUSIONS: Second trimester maternal serum biomarkers may be useful indicators for fetal evaluation for CHD to screen positive pregnancies without identified chromosomal defects.

Bioinformatics identification and validation of biomarkers and infiltrating immune cells in endometriosis.

Background: Endometriosis (EM) is a common gynecological disorder that often leads to irregular menstruation and infertility. The pathogenesis of EM remains unclear and delays in diagnosis are common. Thus, it is urgent to explore potential biomarkers and underlying molecular mechanisms for EM diagnosis and therapies. Methods: Three EM-related datasets (GSE11691, GSE25628, and GSE86534) were downloaded from the Gene Expression Omnibus (GEO) which were integrated into a combined dataset after removing batch effect. Differentially expressed immune cell-related genes were obtained by CIBERSORT, WGCNA, and the identification of differentially expressed genes. Random forest model (RF), support vector machine model (SVM), and generalized linear model (GLM) were then constructed and the biomarkers for EM were determined. A nomogram evaluating the risk of disease was constructed and the validity was assessed by the calibration curve, DCA curve, and clinical impact curve. Single-gene Gene Set Enrichment Analysis (GSEA)was performed to explore the molecular mechanisms of biomarkers. The ceRNA regulatory network of biomarkers was created by Cytoscape and potential target drugs were obtained in the DGIdb database (Drug-Gene Interaction database).The expression levels of biomarkers from clinical samples was quantified by RT-qPCR. Results: The ratio of eight immune cells was significantly different between the eutopic and ectopic endometrium samples. A total of eight differentially expressed immune cell-related genes were investigated. The SVM model was a relatively suitable model for the prediction of EM and five genes (CXCL12, PDGFRL, AGTR1, PTGER3, and S1PR1) were selected from the model as biomarkers. The calibration curve, DCA curve, and clinical impact curve indicated that the nomogram based on the five biomarkers had a robust ability to predict disease. Single gene GSEA result suggested that all five biomarkers were involved in labyrinthine layer morphogenesis and transmembrane transport-related biological processes in EM. A ceRNA regulatory network containing 184 nodes and 251 edges was constructed. Seven drugs targeting CXCL12, 49 drugs targeting AGTR1, 16 drugs targeting PTGER3, and 21 drugs targeting S1PR1 were extracted as potential drugs for EM therapy. Finally, the expression of PDGFRL and S1PR1 in clinical samples was validated by RT-qPCR, which was consistent with the result of public database. Conclusions: In summary, we identified five biomarkers (CXCL12, PDGFRL, AGTR1, PTGER3, and S1PR1) and constructed diagnostic model, furthermore predicted the potential therapeutic drugs for EM. Collectively, these findings provide new insights into EM diagnosis and treatment.

Struma Ovarii Associated with Ascites and Elevated CA125: Two Case Reports and Review of the Literature.

Struma ovarii is a rare variety of specialized monodermal mature ovarian teratoma, it is composed predominantly of thyroid tissue. Ascites is present in one third of patients. The combination of struma ovarii, marked ascites and elevated CA125 is a rare condition, which may mimic ovarian cancer. We described two cases presenting with pelvic mass, ascites and elevated serum CA125 levels, frozen section and final pathology turned out to be struma ovarii. Ascites disappeared and the level of CA125 returned to normal level after operation. One of the cases was associated with pleural effusion, leading to a condition called pseudo-Meigs' syndrome. Then we reviewed the related literatures to explore the possible mechanism of ascites and pleural effusion, the reason of CA125 elevation and imaging manifestations of struma ovarii. In conclusion, struma ovarii should be considered in the differential diagnosis preoperatively, when presented with pelvic mass, ascites and an elevated CA125 level.

Time Intervals between Double Primary Breast and Ovarian Cancers and Survival Outcomes of Patients with Both Cancers: A SEER Database Analysis.

Background: The time interval rules and survival outcomes of individuals with synchronous and metachronous breast cancer (BC) and ovarian cancer (OC) were examined in this retrospective population-based investigation. Methods: The National Cancer Institute's Surveillance, Epidemiology, and End Results database was used to create a cohort of people diagnosed with BC and OC between 1973 and 2015. Patients were separated into three groups: those with main BC followed by primary OC (group 1), those with synchronous primary breast and ovarian cancer (group 2), and those with OC prior to BC (group 3). The Kaplan-Meier technique was used to assess overall survival (OS) and cancer-specific survival (CSS). Results: A total of 4,975 patients were identified: 2,929 patients in group 1, 680 patients in group 2, and 1,366 patients in group 3. The average duration between these tumors was 60 months (range 0-499). Approximately 50% of second primary cancer cases occurred during the first 60 months of the first primary cancer diagnosis, and more than 70% occurred within the first 120 months. The median survival time for 4,975 individuals was 140 months. Group 2 had the smallest median OS (35 months), whereas group 3 had the longest (45 months) (239 months). Conclusions: The majority of second primary cancer cases occurred during the first 120 months following the diagnosis of the first original malignancy. Individuals who had primary OC prior to BC had better prognoses, whereas patients who had synchronous BC and OC had worse prognoses.