Normal Uterine Tissue Research Articles

Post-Uterine Artery Embolization: 3-Day MRI Changes and Their Predictive Value for Therapeutic Efficacy in Symptomatic Uterine Fibroids.

To summarize and discuss3-days MRI changes after uterine artery embolization (UAE) and their predictive value for efficacy. From August 2016 to April 2023, 52 patients underwent enhanced MRI within 3 days post-embolization. We retrospectively analyzed clinical and imaging data, focusing on MR characteristics at the 3-day mark, comparing pre- and post-embolization images. Patients were categorized based on 3-day MR findings into complete and incomplete necrosis groups, with clinical efficacy compared over 6 months. Our study included 30 cases of multiple leiomyomas and 22 of single leiomyomas. Postoperative MRI revealed complete necrosis in 31 tumors and incomplete necrosis in 21 tumors. At 3 days, MR ADC imaging showed increased signals in necrotic areas, mildly increased signals on T2-weighted images, and minimal changes on T1-weighted images. Six-month follow-up showed no significant difference in symptom improvement between groups (p = 0.524, p = 0.587, p = 0.615). However, a significant difference was found in leiomyoma volume reduction, with 70.63 ± 15.53% in the complete necrosis group and 51.36 ± 25.20% in the incomplete necrosis group (p＜0.001), highlighting the impact of necrosis extent on volumetric reduction. Short-term MRI changes after UAE can reflect changes in blood supply to fibroids and normal uterine tissue, and have good predictive value for medium-term embolization efficacy. This study describes short-term MR manifestations of complete and incomplete embolism, aiding in predicting long-term outcome.

Analysis of Expression of the ANG1, CaSR and FAK Proteins in Uterine Fibroids.

Understanding the molecular factors involved in the development of uterine myomas may result in the use of pharmacological drugs instead of aggressive surgical treatment. ANG1, CaSR, and FAK were examined in myoma and peripheral tissue samples taken from women after myoma surgery and in normal uterine muscle tissue samples taken from the control group. Tests were performed using tissue microarray immunohistochemistry. No statistically significant differences in ANG1 expression between the tissue of the myoma, the periphery, and the normal uterine muscle tissue of the control group were recorded. The CaSR value was reduced in the myoma and peripheral tissue and normal in the group of women without myomas. FAK expression was also lower in the myoma and periphery compared to the healthy uterine myometrium. Calcium supplementation could have an effect on stopping the growth of myomas.

Examining the Mechanism of Treatment for Primary Dysmenorrhea with Wenjing Huoxue Decoction based on Transcriptomics, Metabolomics, and Network Pharmacology.

Wenjing Huoxue Decoction (WJHXD) is a traditional treatment for primary dysmenorrhea (PD) that can quickly relieve various symptoms caused by PD. Previous clinical studies have shown that WJHXD has better long-term efficacy than ibuprofen in the treatment of PD and can reverse the disorder of T cell subsets. To investigate the effect of WJHXD on serum-related factors in the treatment of PD, including the identification of key targets, pathways, and active ingredients. In order to study the effects of the WJHXD intervention in Parkinson's Disease (PD) rats, we used transcriptomics and metabolomics methods to examine the differentially expressed genes (DEGs) and differentially expressed metabolites (DEMs). We also utilized network pharmacology to predict the target and effective route of WJHXD in treating PD. Finally, we employed molecular docking (MD) technology to confirm the placement of important targets and metabolites. WJHXD has been found to be effective in prolonging the onset time and decreasing the number of writhing episodes in PD rats after oxytocin injection. It has also been observed to reduce the levels of PGF2, COX-2, AVP, and PGE2 in the serum of PD rats to different degrees. Transcriptomics analysis has revealed that the core targets of WJHXD include KRT1, KRT16, CCL5, F2, NOS2, RAC2, and others, while the core pathways are Calcium signaling and cAMP signaling. The Estrogen signaling pathway was found to be downregulated in PD rats compared to normal uterine tissue, but WJHXD was able to up-regulate the pathway. A combined transcriptomics and metabolomics analysis suggested that WJHXD may be involved in eight metabolism-related pathways, with the most reliable ones being mucin-type O-glycan biosynthesis and glycolysis or gluconeogenesis. MD has shown that Hydroxyisocaproic acid may bind to important targets such as SLC6A4, PTGER3, IGFBP3, and IGF2. In WJHXD, the most targeted herbs were Corydalis rhizoma, licorice, and Myrrha. The most targeted active ingredients include quercetin, 3'-Hydroxy-4'-O-methylglabridin, shinpterocarpin, and isorhamnetin. Potential targets include PTGS2, NOS2, AR, SCN5A, and GAS6. Analysis revealed 72 highly reliable relationships between group A and B DEGs and DEMs, with 23 positive correlations and 49 negative correlations among them. A combined analysis of transcriptomics, metabolomics, and network pharmacology was used to identify possible targets, pathways, and active ingredients of WJHXD in PD treatment, and the correlation between DEGs and DEMs was investigated. However, further research is required to confirm the relationship between active ingredients, targets, and metabolites.

Constructing a Classification Model for Cervical Cancer Tumor Tissue and Normal Tissue Based on CT Radiomics.

This study aimed to develop an automated classification framework for distinguishing between cervical cancer tumor and normal uterine tissue, leveraging CT images for radiomics feature extraction. We retrospectively analyzed CT images from 117 cervical cancer patients. To distinguish between cancerous and healthy tissue, we segmented gross tumor volume and normal uterine tissue as distinct regions of interest (ROIs) using manual segmentation techniques. Key radiomic parameters were extracted from these ROIs. To bolster model's predictive capability, the data was stratified into train data (70%) and validation data (30%). During feature selection phase, we applied Least Absolute Shrinkage and Selection Operator regression algorithm to identify most relevant features. Subsequently, we built classification models using five state-of-the-art machine learning algorithms: Support Vector Machine (SVM), Random Forest (RF), K-Nearest Neighbors (KNN), Extreme Gradient Boosting (XGBoost), and Decision Tree (DT). Ultimately, the performance of each model was evaluated. Through stringent feature selection process, we identified 18 pivotal radiomic features for classification of cervical cancer and normal uterine tissue. When applied to test data, all five models achieved excellent performance, with area under the curve (AUC) values ranging from 0.8866 to 0.9190 (SVM: 0.9144, RF: 0.9078, KNN: 0.9051, DT: 0.8866, XGBoost: 0.9190), all surpassing threshold of 0.8. In terms of test data, all five models had high sensitivity; accuracy of SVM, RF, and XGBoost models was comparable; and specificity of five models was similar. XGBoost model outperformed the others in terms of diagnostic accuracy, achieving an AUC of 0.8737 (95% CI: 0.8198-0.9277) for train data and 0.9190 (95% CI: 0.8525-0.9854) for test data. Our findings underscore the potential of CT radiomics combined with machine learning algorithms for accurately classifying cervical cancer tumors and normal uterine tissue with high recognition capabilities. This approach holds significant promise for clinical diagnostics.

Clinical Characteristics and Local Histopathological Modulators of Endometriosis and Its Progression.

This retrospective study included 243 patients from whom tissue with E/A or normal control uterine tissue was harvested and stained by histochemical and classical immunohistochemical staining. We assessed the symptomatology of the patients, the structure of the ectopic epithelium and the presence of neovascularization, hormone receptors, inflammatory cells and oncoproteins involved in lesion development. Atypical areas were analyzed using multiple immunolabeling techniques. The cytokeratin (CK) CK7+/CK20- expression profile was present in E foci and differentiated them from digestive metastases. The neovascularization marker cluster of differentiation (CD) 34+ was increased, especially in areas with malignant transformation of E or A foci. T:CD3+ lymphocytes, B:CD20+ lymphocytes, CD68+ macrophages and tryptase+ mast cells were abundant, especially in cases associated with malignant transformation, being markers of the proinflammatory microenvironment. In addition, we found a significantly increased cell division index (Ki67+), with transformation and inactivation of tumor suppressor genes p53, B-cell lymphoma 2 (BCL-2) and Phosphatase and tensin homolog (PTEN) in areas with E/A-transformed malignancy. Proinflammatory/vascular/hormonal changes trigger E/A progression and the onset of cellular atypia and malignant transformation, exacerbating symptoms, especially local pain and vaginal bleeding. These triggers may represent future therapeutic targets.

A differential diagnosis between uterine leiomyoma and leiomyosarcoma using transcriptome analysis

BackgroundThe objective of this study was to estimate the accuracy of transcriptome-based classifier in differential diagnosis of uterine leiomyoma and leiomyosarcoma. We manually selected 114 normal uterine tissue and 31 leiomyosarcoma samples from publicly available transcriptome data in UCSC Xena as training/validation sets. We developed pre-processing procedure and gene selection method to sensitively find genes of larger variance in leiomyosarcoma than normal uterine tissues. Through our method, 17 genes were selected to build transcriptome-based classifier. The prediction accuracies of deep feedforward neural network (DNN), support vector machine (SVM), random forest (RF), and gradient boosting (GB) models were examined. We interpret the biological functionality of selected genes via network-based analysis using GeneMANIA. To validate the performance of trained model, we additionally collected 35 clinical samples of leiomyosarcoma and leiomyoma as a test set (18 + 17 as 1st and 2nd test sets).ResultsWe discovered genes expressed in a highly variable way in leiomyosarcoma while these genes are expressed in a conserved way in normal uterine samples. These genes were mainly associated with DNA replication. As gene selection and model training were made in leiomyosarcoma and uterine normal tissue, proving discriminant of ability between leiomyosarcoma and leiomyoma is necessary. Thus, further validation of trained model was conducted in newly collected clinical samples of leiomyosarcoma and leiomyoma. The DNN classifier performed sensitivity 0.88, 0.77 (8/9, 7/9) while the specificity 1.0 (8/8, 8/8) in two test data set supporting that the selected genes in conjunction with DNN classifier are well discriminating the difference between leiomyosarcoma and leiomyoma in clinical sample.ConclusionThe transcriptome-based classifier accurately distinguished uterine leiomyosarcoma from leiomyoma. Our method can be helpful in clinical practice through the biopsy of sample in advance of surgery. Identification of leiomyosarcoma let the doctor avoid of laparoscopic surgery, thus it minimizes un-wanted tumor spread.

Signatures of necroptosis-related genes as diagnostic markers of endometriosis and their correlation with immune infiltration

BackgroundEndometriosis (EMS) occurs when normal uterine tissue grows outside the uterus and causes chronic pelvic pain and infertility. Endometriosis-associated infertility is thought to be caused by unknown mechanisms. In this study, using necroptosis-related genes, we developed and validated multigene joint signatures to diagnose EMS and explored their biological roles.MethodsWe downloaded two databases (GSE7305 and GSE1169) from the Gene Expression Omnibus (GEO) database and 630 necroptosis-related genes from the GeneCards and GSEA databases. The limma package in Rsoftware was used to identify differentially expressed genes (DEGs). We interleaved common differentially expressed genes (co-DEGs) and necroptosis-related genes (NRDEGs) in the endometriosis dataset. The DEGs functions were reflected by gene ontology analysis (GO), pathway enrichment analysis, and gene set enrichment analysis (GSEA). We used CIBERSORT to analyze the immune microenvironment differences between EMS patients and controls. Furthermore, a correlation was found between necroptosis-related differentially expressed genes and infiltrating immune cells to better understand the molecular immune mechanism.ResultsCompared with the control group, this study revealed that 10 NRDEGs were identified in EMS. There were two types of immune cell infiltration abundance (activated NK cells and M2 macrophages) in these two datasets, and the correlation between different groups of samples was statistically significant (P < 0.05). MYO6 consistently correlated with activated NK cells in the two datasets. HOOK1 consistently demonstrated a high correlation with M2 Macrophages in two datasets. The immunohistochemical result indicated that the protein levels of MYO6 and HOOK1 were increased in patients with endometriosis, further suggesting that MYO6 and HOOK1 can be used as potential biomarkers for endometriosis.ConclusionsWe identified ten necroptosis-related genes in EMS and assessed their relationship with the immune microenvironment. MYO6 and HOOK1 may serve as novel biomarkers and treatment targets in the future.

Effects ofCallicarpa nudifloraGranules on the Proliferation and Apoptosis of Uterine Fibroid Cells

This research was aimed to discuss and understand the effects and mechanisms of action ofCallicarpa nudifloragranules on proliferation and apoptosis of uterine leiomyoma (UL) cells. Firstly, normal uterine myometrium (UM) and UL tissues were collected, and the levels of p-Akt and Phosphatase and Tensin Homolog (PTEN) in UL tissues were detected using immunohistochemistry. Next, the UL cells were successfully obtained using enzymatic digestion, and their identification was performed using alpha-smooth muscle actin (α-actin) immunocytochemistry. Specifically, the cells were grouped into four: a control group (CG), a low-dose group (LDG, 50 mg/LCallicarpa nudiflorasolution), a medium-dose group (MDG, 100 mg/LCallicarpa nudiflorasolution), and a high-dose group (HDG, 200 mg/LCallicarpa nudiflorasolution). Moreover, the proliferation of UL cells was assessed using the thiazolyl blue (MTT) assay, while cell apoptosis was analyzed using flow cytometry (FCT). Real-time fluorescent quantitative PCR (fq-PCR) and Western blot assay (WBA) were utilized to determine the PAI-1, P38, TGF-β1, E-cadherin, and Vimentin in UL cells. The results revealed that the positive rate (PR) of p-Akt in the UL tissues was much higher to that in normal UM tissues (P&lt; 0.001). More than 90% of UL cells were positive forα-actin. The viabilities of UL cells in theCallicarpa nudifloratreatment groups were greatly weakened to that of untreated cells (P&lt; 0.05). Viability of UL cells in the HDG group was the lowest, showing a great difference withP&lt; 0.01 to the LDG group and that withP&lt; 0.05 to the MDG group, while that between the MDG and LDG groups exhibited a great difference withP&lt; 0.05. AR of UL cells in CG group was sharply lower to that in theCallicarpa nudifloratreatment groups, showing great differences withP&lt; 0.05,P&lt; 0.01, andP&lt; 0.001, respectively. AR of UL cells in HDG group was higher to the LDG group (P&lt; 0.01) and MDG group (P&lt; 0.05), and that in LDG group was lower and exhibited a great difference withP&lt; 0.05 to the MDG group. The HDG, LDG, and MDG groups exhibited greatly lower TGF-β1, PAI-1, and P38 to the CG group (P&lt; 0.05). In the HDG group, the TGF-β1, PAI-1, P38, and Vimentin levels were greatly lower and presented a great difference withP&lt; 0.01 to those in the CG group and LDG group. Additionally, E-cadherin in UL cells was elevated in the LDG and MDG groups to CG group, showingP&lt; 0.05 andP&lt; 0.01, respectively. Such findings indicated that theCallicarpa nudifloragranules can suppress proliferation of UL cells and promote their apoptosis, which may be associated with the TGF-β1/P38/PAI-1 singling pathway (SPW).

Role of protein tyrosine phosphatase receptor type M in epithelial ovarian cancer progression

BackgroundEpithelial ovarian cancer (EOC) is often diagnosed at advanced stages with low survival rates. Protein tyrosine phosphatase receptor type M (PTPRM) is involved in cancer development and progression; however, its role in EOC remains unclear. In this study,we aimed to detect PTPRM expression in ovarian epithelial tumors, analyze its relationship with the clinicopathological features and survival prognosis of patients with EOC, and provide a theoretical basis for new targets for EOC treatment. Fifty-seven patients with EOC treated at our hospital between January 2012–January 2014 were included; along with 18 borderline and 30 benign epithelial ovarian tumors and 15 normal ovarian and uterine tube tissue samples from patients surgically treated at our hospital during the same period. PTPRM expression was immunohistochemically detected, and we analyzed its relationship with clinicopathological features and prognosis. Associations between PTPRM expression and survival prognosis of patients with EOC were analyzed using the Gene Expression Profiling Interactive Analysis (GEPIA) and Kaplan–Meier Plotter databases.ResultsPTPRM had the highest expression rates in normal ovarian and uterine tube tissues, followed by benign and borderline epithelial ovarian tumors; the lowest positive expression rate was observed in EOC tumors. PTPRM expression differed significantly among groups (P < 0.05). The positive PTPRM expression rate significantly decreased with age, progressing clinical stage, and tumor recurrence, and the larger the mass diameter, the higher the positive PTPRM expression rate. PTPRM expression was significantly lower in ovarian cancer compared with that in normal tissues in the GEPIA database (P < 0.05). The overall survival (OS) and disease-free survival(DFS) rates were higher in the PTPRM high-expression group, with statistically significant (P < 0.05) and insignificant (P > 0.05) differences, respectively. The OS rate of the high-expression group compared with the low-expression group in the Kaplan–Meier Plotter database was higher, although without statistical significance (P > 0.05), and progression-free survival(PFS) was higher with statistical significance (P < 0.05).ConclusionPTPRM expression was low in patients with EOC, and the PTPRM positive-expression rate significantly decreased with progressing stages of EOC and tumor recurrence, suggesting that PTPRM acts as a tumor suppressor in EOC progression. Negative PTPRM expression may predict poor clinical outcomes in patients with EOC.

Study of the unique cellular molecular characteristics of moderately intrauterine adhesion based on single-cell RNA sequencing

To depict the cell landscape and molecular biological characteristics of human intrauterine adhesion (IUA) so as to better understand its immune microenvironment and provide new inspirations for clinical treatment. Four patients with IUA who underwent hysteroscopic treatment at Dongguan Maternal and Child Health Care Hospital from February 2022 to April 2022 were selected as the study subjects. Hysteroscopy was used to collect the tissues of IUA, which were graded based on the patient's medical history, menstrual history and status of IUA. Library construction, sequencing, single cell data comparison and gene expression matrix construction were carried out in strict accordance with the single cell RNA sequencing process. Thereafter, the UMAP dimension reduction analysis of cell population and genetic analysis were carried out based on the cell types. A total of 27 511 cell transcripts were obtained from four moderately graded IUA tissue samples and assigned to six cell lineages including T cells, mononuclear phagocytes, epithelial cells, fibroblasts, endothelial cells and erythrocytes. Compared with normal uterine tissue cells, the four samples showed different cell distribution, and the proportions of mononuclear phagocytes and T cells in sample IUA0202204 were significantly increased, suggesting a strong cellular immune response. The cell diversity and heterogeneity of moderate IUA tissues have been described. Each cell subgroup has unique molecular characteristics, which may provide new clues for further study of the pathogenesis of IUA and heterogeneity among the patients.

Human LY6 gene family: potential tumor-associated antigens and biomarkers of prognosis in uterine corpus endometrial carcinoma.

The human Lymphocyte antigen-6 (LY6) gene family has recently gained interest for its possible role in tumor progression. We have carried out in silico analyses of all known LY6 gene expression and amplification in different cancers using TNMplot and cBioportal. We also have analyzed patient survival by Kaplan-Meier plotter after mining the TCGA database. We report that upregulated expression of many LY6 genes is associated with poor survival in uterine corpus endometrial carcinoma (UCEC) cancer patients. Importantly, the expression of several LY6 genes is elevated in UCEC when compared to the expression in normal uterine tissue. For example, LY6K expression is 8.25× higher in UCEC compared to normal uterine tissue, and this high expression is associated with poor survival with a hazard ratio of 2.42 (p-value = 0.0032). Therefore, some LY6 gene products may serve as tumor-associated antigens in UCEC, biomarkers for UCEC detection, and possibly targets for directing UCEC patient therapy. Further analysis of tumor-specific expression of LY6 gene family members and LY6-triggered signaling pathways is needed to uncover the function of LY6 proteins and their ability to endow tumor survival and poor prognosis in UCEC patients.

Myoma Uteri: Stiffness from MR Elastography in Thai Patients: A Pilot Study

Objective: To study the feasibility of magnetic resonance elastography (MRE) for the evaluation of myoma uteri in Thai patients. Materials and Methods: A prospective descriptive study was conducted between September 2020 and October 2021. Twenty-six myoma uteri patients were performed in a 3.0-Tesla MRI scanner, added with a pneumatic system generating mechanical waves. For the MRE assessment, the number of acquired slices, mean stiffness with standard deviation (SD), minimum, and maximum stiffnesses (kPa) were recorded. Results: The mean stiffness of the myoma uteri was 3.02 kPa (range 1.83 to 5.06, SD 0.79). Their volume ranged from 56.24 to 716.35 cm³ (mean 237.74, SD 187.63. The proportion of lesions ranged from 31.87% to 74.36% of the whole uterus (mean 44.94, SD 16.98). Conclusion: Uterine MRE is a feasible diagnostic tool for studying the tissue stiffness of the myoma uteri, which is higher than normal uterine tissue. The mean stiffness of Thai patient myoma uteri was lower than other ethnicities. Keywords: Myoma uteri; Uterine fibroid; Uterine leiomyoma; MRE; Tissue stiffness

Abstract 5698: Genome-wide DNA methylation profiling for differentiating synchronous endometrial and ovarian cancer

Abstract Background: Synchronous endometrial and ovarian cancer (SEOC) refers to simultaneously discovered endometrial and ovarian tumours on the same patient. SEOC may represent double primary (DP) tumours that arise independently on the two sites, or clonally related (CR) tumours that arise either on the endometrium or ovary and metastasized to the other site. Accurate differential diagnosis between these two scenarios can implicate on the treatment options and prognostication of SEOC. However, it is difficult to achieve as the histopathological features of DP and CR tumours are often ambiguous. DNA methylation landscape of SEOC may provide markers for differentiating DP and CR tumours but remain largely unexplored. Methods: The archival tissues of six pairs of SEOC and their accompanied normal uterine tissues were retrieved. The tumours were accessed by conventional histopathological criteria and mitochondrial DNA (mtDNA) sequencing to categorize them into DP or CR. We then analyzed the methylation profiles of &amp;gt;850,000 CpG sites of the tumor samples using the Illumina Infinium MethylationEPIC BeadChip array. Downstream methylation analysis was performed in the R/Bioconductor package ChAMP. Gene ontology (GO) enrichment analysis on the methylation driven genes was done using PANTHER. Results: mtDNA sequencing revealed that among the 6 cases of SEOC, there are 3 DP and 3 CR. Clustering analysis of differentially methylated CpGs (DMPs) between tumours of the same tissue type effectively distinguished endometrial and ovarian tumours into DP or CR. Thus, DP and CR endometrial tumour showed 50 DMPs and 15 differentially methylated regions (DMRs), but no significantly enriched signaling pathway was identified. On the other hand, there were 1,105 DMPs and 44 DMRs that mapped to 601 known genes between DP and CR ovarian tumours. GO analysis revealed that the hypomethylated genes in CR ovarian tumour were enriched in several metastatic-related processes, including cellular response to stimulus, regulation of cell projection organisation, cell migration and negative regulation of voltage-gated potassium channel activity. Conclusions: Taken together, our findings demonstrated that DNA methylation profiling can be used for SEOC patient stratification and may enhance our understanding of epigenetic regulation of SEOC pathogenesis. Citation Format: Yusanne Yung Sing Chan, Oscar Gee Wan Wong, Annie Nga Yin Cheung. Genome-wide DNA methylation profiling for differentiating synchronous endometrial and ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5698.

Upregulated Ly-6 gene expression is associated with poor overall survival in uterine corpus endometrial carcinoma patients

Abstract The human Ly-6 (hLy-6) gene family has recently gained interest for its possible role in tumor progression. High expression of hLy-6 genes is associated with poor overall survival in pancreatic ductal adenocarcinoma patients. To expand upon this work, we have carried out in silico analyses of all known hLy-6 gene expression in other cancers using TNMplot which shows differential gene expression in cancer tissues. We have also analyzed the patient survival by Kaplan-Meier plotter after mining the TCGA database. We report that upregulated expression of several hLy-6 genes is associated with poor cancer patient survival in many other cancer types, especially uterine corpus endometrial carcinoma (UCEC). Importantly, the expression of a number of hLy-6 genes is elevated in UCEC tissue when compared to the expression in normal uterine tissue. Further analysis of tumor-specific expression of the hLy-6 gene family is needed to uncover the function of Ly-6 proteins as well as the signaling pathways these proteins trigger that endow tumor survival and poor patient prognosis. Human Ly-6 gene products can potentially be used as biomarkers for cancer detection and as tumor associated antigens to identify high-risk cancer patients and appropriately design and/or calibrate cancer treatments for such patients.

An integrated method for contained laparoscopic specimen removal

Efforts at minimizing pain following minimally invasive (MIS) hysterectomy were hampered by the absence of simple, reliable methods to extract specimens following the 2014 FDA safety warning discouraging power morcellation. To address this gap, an integrated method intended to provide reliable specimen containment with a multi-layer bag, segmentation using bipolar energy, and tissue extraction during laparoscopic gynecological surgical procedures was developed. This report details the pre-clinical studies demonstrating specimen containment integrity, and thermal and electrical safety of the Eximis™ CS (Contained Segmentation) Procedure. All experiments were performed in pigs and approved by the research facility's institutional animal care committee and met USDA and AAALAC guidelines. The study design was composed of three experimental groups; Bovine tongue muscle to simulate normal uterine tissue, bovine tendon draped in a “sausage casing” to simulate hard-thick myoma tissues, and a polymer block wrapped with bacon strips to simulate a severely calcified fibroid. Additionally, there were two control groups: one negative and one positive (heat gun). Each experimental group was tested in four Eximis Capture Bags (8 cuts each) for a total of 24 electromechanical cuts. Thermal evaluation of the small bowel adjacent to the capture bag during device activation was performed with a Micronor FOTEMP4-PLUS-P0-V-B fiberoptic thermometry system. Specimens were then harvested for histology and graded by a blinded pathologist using the Histopathology Scoring Criteria (edema/necrosis/inflammation); 0-4; none to marked. Electrical testing evaluated high frequency electrical leakage current by an independent laboratory with the electrosurgical generator set to 200 W output power, the maximum output setting allowable. Finally, Eximis Capture Bags were tested after segmentation using a proprietary leak test that can detect leaks < a 1 μm hole size. The test group simulating normal uterine tissue (bovine tongue) had small bowel histopathology not different from the negative control group. The calcified fibroid and myoma test groups demonstrated statistically more mucosal edema than the negative control group, but differences were not observed in the submucosa or muscularis layers that were closer to the containment bag. The Eximis Capture Bag measured leakage current directly through the bag of 1.01 mA well below the maximum allowable standard of 150 mA. No evidence of leaks was detected. These studies demonstrate that the Eximis CS is thermally and electrically safe to surrounding tissues and that tissue segmentation and extraction do not impact Eximis Capture Bag’s integrity. Clinical evaluation during human and robotic gynecologic procedures for benign disease is warranted. Additionally, these results suggest the system may be beneficial in other MIS procedures where large specimens are removed.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Abstract GS2-09: Tamoxifen instigates uterine cancer development by activating PI3K signaling and supersedes PIK3CA driver mutations

Abstract Tamoxifen is widely used in the adjuvant treatment of estrogen receptor–positive (ER+) breast cancer and is an important drug for pre-menopausal women and post-menopausal patients who cannot tolerate aromatase inhibitors. Despite the clear clinical benefit in improving relapse-free and overall survival in these patients, an adverse effect of tamoxifen is a 2- to 7-fold increased risk of uterine cancer (UC) after 2-5 years of treatment. To date, the mechanism of tamoxifen-driven tumorigenesis is not well understood, and preventive approaches are lacking. Here, to molecularly characterize tamoxifen-associated uterine cancers (TA-UCs) and gain insights into their unique evolution, we performed whole-exome sequencing of 21 TA-UCs (discovery cohort) and droplet digital PCR (ddPCR) of an additional 40 TA-UCs (validation cohort) obtained from the ‘Tamoxifen Associated Malignancies: Aspects of Risk’ (TAMARISK) study. In addition, we used in vivo mouse models to: (i) further investigate tamoxifen-activated molecular pathways that may be involved in TA-UC tumorigenesis; and (ii) offer mechanistic insights. Overall, we discovered that TA-UCs were genomically similar to non–TA-UCs from The Cancer Genome Atlas (TCGA) project, with one profound exception: TA-UCs are characterized by a lower-than-expected frequency of mutations in two highly prevalent UC driver genes in the PI3K pathway: PIK3CA (14% [3/21] vs 48% [265/554] in non–TA-UC; P=0.003, Fisher’s exact test; Q=0.02, Benjamini-Hochberg FDR) and PIK3R1 (0%, [0/21] vs 31% [174/554]; P=0.001; Q=0.01). We used ddPCR in the independent TA-UC validation cohort and confirmed the low frequency of mutations in PIK3CA (7.5% [3/40] vs 21% [144/685] in control UCs from the Dana-Farber contribution to the AACR GENIE project; P=0.04). We next performed mouse in vivo studies and demonstrated that tamoxifen activated the PI3K pathway and increased cell proliferation in normal mouse uterine tissue through paracrine and autocrine effects, both of which were abrogated by the PI3K inhibitor alpelisib. Taken together, we describe a distinct and novel pathway of carcinogenesis in which tamoxifen acts as a driver event in the uterus and promotes tumor development in a mutation-independent manner. Indeed, tamoxifen may increase the risk of UC by activating the PI3K pathway, which can substitute for the early acquisition of oncogenic PIK3CA or PIK3R1 mutations observed in non–TA-UC tumors. Furthermore, the ability of a PI3K inhibitor to reduce cell proliferation in our mouse model raises the possibility that downregulating the PI3K pathway may prevent or significantly reduce TA-UC development, offering a potential future therapeutic and prevention strategy for specific high-risk patients undergoing tamoxifen therapy. Citation Format: Kirsten Kubler, Agostina Nardone, Shankara Anand, Daniel Gorvich, Marjolein Droog, Francisco Hermida-Prado, Tara Akshi, Avery S Feit, Gabriella Cohen, Gwen Dackus, Matthew Pun, Yanan Kuang, Justin Cha, Mendy Miller, William J Gibson, Cloud P Paweletz, Eliezer M Van Allen, Flora E van Leeuwen, Petra Nederlof, Harry Hollema, Quang-Dé Nguyen, Marian JE Mourits, Ignaty Leshchiner, Chip Stewart, Ursula A Matulonis, Wilbert Zwart, Yosef E Maruvka, Gad Getz, Rinath Jeselsohn. Tamoxifen instigates uterine cancer development by activating PI3K signaling and supersedes PIK3CA driver mutations [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr GS2-09.

Components of NOTCH Signaling for Uterine Cancer Patients' Prognosis.

New molecular biomarkers that could have an independent prognostic value in endometrial cancer are currently under investigation. Recently, it was suggested that genetic changes in the Notch signaling pathway could be associated with the development of endometrial carcinoma. This study aimed to determine the expression of the Notch signaling pathway components in tumour and adjacent normal uterine tissue and to evaluate their importance for the survival of uterine cancer patients. The present study was performed on uterine body samples collected from 109 patients and paired adjacent noncancerous endometrial tissue samples. Kaplan–Meier curves and Cox regression were used for survival analyses. Expression alterations of NOTCH2, NOTCH3, NOTCH4, JAG2, and HES1 were evaluated as independent and significant prognostic factors for uterine cancer patients.

APOBEC mediated mutagenesis drives genomic heterogeneity in endometriosis.

Endometriosis is a benign gynecologic condition, acting as a precursor of certain histological subtypes of ovarian cancers. The epithelial cells of endometriotic tissues and normal uterine endometrium accumulated somatic mutations in cancer-associated genes such as phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) and Kirsten rat sarcoma (KRAS) proto-oncogene. To determine the genomic characteristic of endometriotic epithelial cells and normal uterine endometrium and to identify the predominant mutational process acting on them, we studied the somatic mutation profiles obtained from whole exome sequencing of 14 endometriotic epithelium and 11 normal uterine endometrium tissues and classified them into mutational signatures. We observed that single base substitutions 2/13 (SBS), attributed to Apolipoprotein B mRNA Editing Enzyme Catalytic Subunit (APOBEC) induced mutagenesis, were significant in endometriotic tissues, but not in the normal uterine endometrium. Additionally, the larger number and wider allele frequency distribution of APOBEC signature mutations, compared to cancer-associated driver mutations in endometriotic epithelium suggested APOBEC mutagenesis as an important source of mutational burden and heterogeneity in endometriosis. Further, the relative risk of enriched APOBEC signature mutations was higher in endometriosis patients who were carriers of APOBEC3A/3B germline deletion, a common polymorphism in East Asians which involves the complete loss of APOBEC3B coding region. Our results illustrate the significance of APOBEC induced mutagenesis in driving the genomic heterogeneity of endometriosis.

Identification of condition-specific biomarker systems in uterine cancer.

Uterine cancer is the fourth most common cancer among women, projected to affect 66,000 US women in 2021. Uterine cancer often arises in the inner lining of the uterus, known as the endometrium, but can present as several different types of cancer, including endometrioid cancer, serous adenocarcinoma, and uterine carcinosarcoma. Previous studies have analyzed the genetic changes between normal and cancerous uterine tissue to identify specific genes of interest, including TP53 and PTEN. Here we used Gaussian Mixture Models to build condition-specific gene coexpression networks for endometrial cancer, uterine carcinosarcoma, and normal uterine tissue. We then incorporated uterine regulatory edges and investigated potential coregulation relationships. These networks were further validated using differential expression analysis, functional enrichment, and a statistical analysis comparing the expression of transcription factors and their target genes across cancerous and normal uterine samples. These networks allow for a more comprehensive look into the biological networks and pathways affected in uterine cancer compared with previous singular gene analyses. We hope this study can be incorporated into existing knowledge surrounding the genetics of uterine cancer and soon become clinical biomarkers as a tool for better prognosis and treatment.

LncRNA APTR Promotes Uterine Leiomyoma Cell Proliferation by Targeting ERα to Activate the Wnt/β-Catenin Pathway.

The molecular mechanisms by which uterine leiomyoma (UL) cells proliferate are unclear. Long noncoding RNA (lncRNA) is reported to participate in the occurrence and development of gynecological cancers. We investigated the molecular mechanisms that lncRNA uses in UL. We found that lncRNA Alu-mediated p21 transcriptional regulator (APTR) showed higher expression in UL tumor tissues compared with that in normal uterine tissues. APTR induced cell proliferation and colony formation both in vitro and in vivo. The JASPAR database showed that APTR was likely interacted with ERα, and these molecules were identified via laser scanning confocal microscopy and RNA immunoprecipitation analysis. To verify the correlation between APTR and ERα, we overexpressed and underexpressed APTR and simultaneously expressed ERα. The results showed that APTR function was suppressed. APTR increased the expressions of the proteins in the Wnt pathway, and inhibiting ERα eliminated these responses. In conclusion, our data suggest that APTR promoted leiomyoma cell proliferation through the Wnt pathway by targeting ERα, suggesting a new role of APTR in the Wnt signaling pathway in UL.