Normal Myometrium Research Articles

Diagnostic value of transvaginal contrast-enhanced ultrasound in identifying benign and malignant endometrial lesions and assessing myometrial invasion.

The objective of this study was to evaluate the diagnostic value of transvaginal contrastenhanced ultrasound (CEUS) in differentiating benign from malignant endometrial lesions and assessing the extent of myometrial invasion. A total of 70 patients who underwent surgery for endometrial lesions at the authors' hospital were selected. Transvaginal ultrasound examination and CEUS were performed for quantitative and qualitative analysis. Based on the CEUS results, an International Federation of Gynecology and Obstetrics (FIGO) disease grade was assigned and compared with pathological findings. Postmenopausal vaginal bleeding is a key clinical manifestation of endometrial carcinoma. Among the patients with endometrial carcinoma, compared with normal myometrium, the lesion areas exhibited a greater rate of rise (defined as enhanced intensity divided by enhancement time) and a shorter half-clearance time (P<0.05). These findings suggest that in endometrial carcinoma, the contrast agent displays a "fast-in/fast-out/hyperenhancement" perfusion pattern. In contrast, the characteristic perfusion pattern for benign endometrial lesions is low enhancement (P<0.05). The diagnostic accuracy of CEUS in detecting myometrial invasion was 88% (22 of 25 cases). Transvaginal CEUS is a practical and effective diagnostic imaging method for distinguishing between benign and malignant endometrial lesions. It can also be used to evaluate the depth of myometrial invasion in patients with early-stage endometrial carcinoma.

The Potential of Transforming Growth Factor-Beta1 (TGFβ1) in Fibrotic Tissue of Leiomyoma Specimens

Background: Fibrosis can define as the development of excessive fibrous tissue in an organ or tissue. In the situation of the uterus, leiomyoma or scar tissue may involve fibrosis state. The main feature of fibrosis in myoma is the existence of disordered smooth muscle cells and collagen that can be recognized via histological examinations in addition to extracellular component deposition. Leiomyoma is the most public uterine disorder that is mainly consists of smooth muscle cells, myo-fibroblast, and an obvious content of extracellular matrix. The myofibroblast in myoma lesion produced an elevated content of ECM. This paper displays the fibrotic circumstantial in leiomyoma and the mechanisms related to deposition of ECM components. Methods: this study involved (50) patients of 18-80 years old women diagnosed with benign uterine fibroid. Tumor samples were attained by consent from women undergoing hysterectomy or myomectomy at the Al Karama Teaching Hospitals in Kut, Wasit Province, Iraq between October 2022 to March 2023. samples from normal and fibroid were subjected to routein histological processing and subsequently stained with routein stains for general description, Masson Trichrome stain, Van Gieson's stain to visualize collagen content and histochemical stain (Alcian Blue-Periodic Acid Schiff) to evaluate the profile of mucins secreted via tumor cells.Normal myometrium and myoma tissue expression of TGF β1 growth factor was compared using immunohistochemical staining. Qupath and Image J programs were used to analysis the content of fibrous tissue within normal and myoma tissue. Results: Microscopical results revealed that myoma have a proliferation of overlap fusiform cells organized in bundles alienated by variable amounts of connective tissue fibers, enclosed peripherally by a pseudo capsule. Special stained sections analysis revealed deposition of extracellular martix components specially collagen fibers that vary within each nodular fibroid (intrafascicular, interfascicular) enclosed myocytes that reflect the state of fibrosis that appears as blue – green colors enclosed in Masson's trichrome stain, while in Van Gieson's stain appears as red color. Proliferative tumor cells were revealed via Alcian Blue-PAS stain in dark blue color. results showed distribution of perivascular fibers in myoma as compared with normal adjacent myometrium, which appears in red color in Van Gieson's stain. Immunostaining recorded the significantly higher intensity of TGF-β in the leiomyoma tissue than in the normal myometrium (P &lt; 0.05). The effects of the TGF-β1 inhibitor significantly differed between normal myometrium and leiomyoma tissue, with a greater decrease in cell survival in the leiomyoma tissue (P &lt; 0.05). Conclusion: highly fibrous tissue contents showed within myoma and perivascular than adjacent myometrium, these fibrous tissues were detected via special colours in both Masson Trichrome and Van Gieson's stains. The fibrous tissue within leiomyoma showed higher intensity to TGF than normal myometrium.

Resection of uterine arteriovenous fistula with temporary occlusion of the internal iliac arteries: Case series and literature review.

Uterine arteriovenous fistula (UAVF) is a rare vascular abnormality that can cause severe and potentially life-threatening hemorrhage. Uterine artery embolization is a common treatment that may affect ovarian and uterine perfusion and cause fertility problems. We herein report our experience treating 2 patients with UAVF who underwent resection after temporary occlusion of both internal iliac arteries. Both patients presented with a large UAVF after incomplete miscarriages in the second trimester. Magnetic resonance imaging revealed a UAVF measuring 3.6 × 2.6 × 2.1 cm over the myometrium of the posterior uterine in case 1, and a UAVF of 7.1 × 4.1 × 4.5 cm was identified in case 2. Uterine arteriovenous fistula, retained products of conception. The patients underwent resection of UAVF with temporary occlusion of the internal iliac arteries and hysteroscopic removal of the retained products of conception. Intraoperative bleeding were minimal. Neither patient exhibited abnormal uterine bleeding at the 6-month follow-up. Follow-up ultrasonography and magnetic resonance imaging showed normal uterine myometrium and endometrium and no residual disease. UAVF resection after temporary occlusion of the internal iliac arteries is a promising treatment approach for UAVF. This technique can reduce intraoperative bleeding and remove the potential hemorrhage-related lesion while preserving fertility.

Whole-exome Sequence Analysis of Gastric-type Adenocarcinoma of the Uterine Cervix and Adjacent Lobular Endocervical Glandular Hyperplasia in the Same Case.

Lobular endocervical glandular hyperplasia (LEGH) may be a precursor lesion of gastric-type adenocarcinoma of the uterine cervix (GAS). However, the genetic mechanisms underlying its carcinogenesis remain unclear. To elucidate the oncogenic process from LEGH to GAS, we compared gene mutations in early-stage GAS and adjacent LEGH in the same case. Fresh-frozen tissue sections were obtained from a patient with Stage IB3 GAS and adjacent LEGH who had undergone hysterectomy. Using laser microdissection, we harvested the LEGH and GAS portions separately from these sections and extracted the genomic DNA. Somatic variant analysis using whole-exome sequencing used DNA from the normal myometrium as a reference sequence. Somatic variants involving amino acid substitutions were detected in 61 and 125 locations in LEGH and GAS, respectively. Seven variants were common in both lesions, of which the pathogenic variant was GNAS only (c.2531G>A, p.R844H), a mutation frequently reported in pancreatic and colorectal cancers. LEGH had no other pathogenic variants; another pathogenic variant in GAS was found only at the same amino acid site as GNAS (c.2530C>T, p.R844C). In the present case, LEGH and GAS shared the same pathogenic variant of GNAS, indicating that both lesions had a common origin. Furthermore, the current results showed that the second GNAS variant is associated with the progression of LEGH to GAS. Further studies are required to elucidate GAS's pathogenesis and biological characteristics.

P-321 Uterine fibroids secrete vasodilatory factors that induce their response via the opioid pathway

Abstract Study question Do uterine fibroids produce vasoactive factors that could affect vasculature and induce fibroid associated symptoms? Summary answer Ex vivo secretions from uterine fibroids induce a vasodilatory response in rat mesenteric arteries that is mediated via the opioid receptor pathway. What is known already Uterine fibroids (leiomyoma) are a common but clinically neglected gynaecological condition, where benign smooth muscle tumours arise from the myometrium and cause heavy menstrual bleeding (HMB), pain and infertility. Fibroids affect up to one in two women of reproductive age and are the main reason for failure of both medical and surgical treatments in women with HMB. Fibroids are proposed to affect the systemic vasculature system, with angiogenic growth factors and abnormal vasculature implicated in fibroid pathophysiology. However, the precise mechanism of fibroid induced HMB has yet to be fully elucidated. Study design, size, duration Fibroid and myometrium biopsies were prospectively collected from 13 patients undergoing MyoSure, myomectomy or hysterectomy for the treatment of HMB associated with fibroids at Liverpool Women’s Hospital, a tertiary referral centre. The collection and use of human tissue was ethically approved, and all participants gave informed written consent. Tissue biopsies were incubated in a basal medium for 24 hours. Conditioned medium was collected and used for wire myography assays to examine vascular reactivity. Participants/materials, setting, methods Second order mesenteric arteries from female Wistar rats were mounted on a wire myograph system and toned with uridine triphosphate (UTP) before being exposed to control (non-conditioned), normal myometrium (n = 4) or fibroid (n = 9) conditioned medium. Vascular activity was measured relative to UTP induced vasoconstriction. To determine the mediatory factors of observed vasoreactivity, inhibitors of well characterised pathways were systematically tested in the presence of conditioned medium. Main results and the role of chance Fibroid conditioned medium caused a marked dilation of UTP induced vasoconstriction in rat mesenteric arteries, which was significantly greater than that induced by non-conditioned, or control myometrium conditioned medium. Blockage of ATP-sensitive potassium (KATP) channels with PNU37883A completely inhibited fibroid induced vasodilation. However, blockage of other potassium channels did not inhibit dilation, suggesting that a KATP-mediated PKA-dependent pathway is being stimulated. To test the potential involvement of endothelial-derived nitric oxide synthase in vasodilation, vessels were treated with the inhibitor L-NAME, which partially blocked fibroid induced vasodilation. The specificity of nitric oxide synthase inhibition was confirmed by treating vessels with acetylcholine or carbachol, which induced vasodilation via activation of nitric oxide synthase. In the presence of L-NAME, the vasodilatory effects of acetylcholine and carbachol were blocked, thus confirming inhibitor specificity. An adenosine receptor inhibitor, theophylline, had no effect on fibroid induced vasodilation, thus ruling out the adenosine pathway. Blockage of bradykinin and adrenergic receptors also failed to prevent fibroid induced dilation in mesenteric arteries. However, blocking opioid receptors with naloxone completely inhibited vasodilation in the presence of fibroid conditioned medium. These findings suggest that fibroid induced vasodilation is mediated via endothelial nitric oxide synthase and an unknown opioid compound. Limitations, reasons for caution This study was performed using excised tissue in an ex vivo environment, which may not fully simulate the fibroid secretome in vivo. Although rat mesenteric arteries are a well-established model, human uterine arteries may be more appropriate to study fibroid induced vasoreactivity. Wider implications of the findings Fibroids are responsible for up to half of all hysterectomies and healthcare costs run into the billions globally. This study demonstrates that fibroid secreted factors can impact vasoreactivity, which has important connotations for disease pathophysiology regarding HMB. Targeting vasoactive compounds produced by fibroids may be a future novel treatment option. Trial registration number NA

Engineered uterine primary myometrial cells with high-mobility group AT-hook 2 overexpression display a leiomyoma-like transcriptional and epigenomic phenotype

To determine if engineered HMGA2 overexpressing uterine primary myometrial cells recapitulate the transcriptional and epigenomic features of HMGA2-subtype leiomyomas. Isolated primary, "normal" myometrial cells from 3 patients were engineered to overexpress HMGA2 to determine how HMGA2 establishes transcriptomic and epigenomic features of HMGA2-overexpressing leiomyoma. Academic research laboratory PATIENTS: Primary myometrial cells were isolated from normal myometrium obtained from three patients undergoing hysterectomy. Not applicable MAIN OUTCOME MEASURES: Determined genome-wide transcriptomic and epigenomic features of engineered HMGA2-overexpressing uterine primary myometrial cells. Engineered HMGA2-V5-overexpressing primary myometrial cells approximated the HMGA2 expression level observed in HMGA2-overexpression subtype leiomyoma. HMGA2-V5 expression resulted in differential expression of 1612 genes (FDR < 0.05) which were found to be enriched in pathways associated with leiomyoma formation, including extracellular matrix organization. Comparative gene expression analysis between HMGA2-V5 engineered primary cells and HMGA2-overexpression subtype leiomyoma revealed significant overlap of differentially expressed genes. Mechanistically, HMGA2-V5 overexpression resulted in 41,323 regions with differential H3K27ac deposition (FDR < 0.05) and 205,605 regions of altered chromatin accessibility (FDR < 0.05). Transcription factor binding site analysis implicated the AP-1 family of transcription factors. • What clinical problem is addressed by these studies? About 10-15% of the uterine leiomyoma cases exhibit increased expression of HMGA2; our study addresses the primary role of HMGA2 overexpression in the pathogenesis of leiomyoma. • What are the key findings? Ectopic expression of HMGA2 in primary myometrial cells transforms transcriptional and epigenomic machinery in these cells which to some extent resemble features of leiomyoma. • How do these findings apply to human fertility or the reproductive process? Our study provides a comprehensive analysis into the changes occurring in epigenome of myometrial cells when HMGA2 is overexpressed. Because epigenome can be targeted with drugs, therefore understanding alterations in epigenome provides new avenues for treating leiomyoma which affects reproductive age women.

Leiomyomatosis: A Rare Case of Mistaken Identity

Abstract: Diffuse uterine leiomyomatosis (DUL) is a rare benign uterine pathology that is often misdiagnosed as uterine leiomyomas. We present a case of a woman with abnormal uterine bleeding treated with multiple medical therapies without symptomatic relief. She underwent two surgeries, almost a year apart, with the histopathologic diagnosis of leiomyomas. After numerous failed attempts at medical and surgical treatments, the patient requested definitive surgery and underwent a robotic-assisted laparoscopic hysterectomy. Histopathologic analysis of the entire uterine specimen confirmed diffuse uterine leiomyomatosis. Case Report: A 31-year-old female was referred to our office with a history of abnormal uterine bleeding and heavy menstrual bleeding who failed hormonal therapies. Levonorgestrel-releasing intrauterine devices were inserted at two different times and were expelled due to heavy menses. Pelvic magnetic resonance imaging reported leiomyomas. The patient underwent hysteroscopy and a robotic-assisted myomectomy. Pathology revealed leiomyomas. Seven months after her first surgery, her bleeding recurred, and the patient became severely anemic, requiring hospitalization and transfusion of 2 units of packed RBCs. Over a year later, the patient underwent a second hysteroscopic and robotic myomectomy after failed medical therapies of tranexamic acid, progesterone medication, and gonadotropin-releasing hormone antagonist with minimal symptom relief. Her quality of life suffered tremendously due to ongoing AUB-HMB. After seven months, the patient requested definitive surgical therapy and underwent a robotic-assisted laparoscopic hysterectomy. Pathology reported diffuse uterine leiomyomatosis. Diffuse uterine leiomyomatosis is a rare benign uterine pathology that is often misdiagnosed as uterine leiomyomas. Early diagnosis of DUL is challenging and often inconclusive. Less than 50 diffuse uterine leiomyomatosis cases have been reported in the literature. The myometrium in DUL is almost entirely replaced by numerous ill-defined benign smooth muscle tumors. MRI offers mapping of the size, site, and distribution of leiomyomas, but findings are difficult to differentiate from DUL. The pathophysiology of DUL development remains poorly understood, with patients frequently complaining of abnormal uterine bleeding, heavy menstrual bleeding, fertility issues, and dysmenorrhea. DUL is often misdiagnosed as leiomyomas or adenomyosis clinically, given the similarity in manifestations. Leiomyomatosis is histologically characterized by the proliferation of smooth muscle cells forming fascicles and nodules, which blend with normal myometrium tissue. Hysterectomy is the definitive treatment of DUL, with this case report demonstrating the close similarity between the two.

Single-cell and spatial transcriptomics reveal alterations in trophoblasts at invasion sites and disturbed myometrial immune microenvironment in placenta accreta spectrum disorders

BackgroundPlacenta accreta spectrum disorders (PAS) are a severe complication characterized by abnormal trophoblast invasion into the myometrium. The underlying mechanisms of PAS involve a complex interplay of various cell types and molecular pathways. Despite its significance, both the characteristics and intricate mechanisms of this condition remain poorly understood.MethodsSpatial transcriptomics (ST) and single-cell RNA sequencing (scRNA-seq), were performed on the tissue samples from four PAS patients, including invasive tissues (ST, n = 3; scRNA-seq, n = 4), non-invasive normal placenta samples (ST, n = 1; scRNA-seq, n = 2). Three healthy term pregnant women provided normal myometrium samples (ST, n = 1; scRNA-seq, n = 2). ST analysis characterized the spatial expression landscape, and scRNA-seq was used to identify specific cellular components in PAS. Immunofluorescence staining was conducted to validate the findings.ResultsST slices distinctly showed the myometrium in PAS was invaded by three subpopulations of trophoblast cells, extravillous trophoblast cells, cytotrophoblasts, and syncytiotrophoblasts, especially extravillous trophoblast cells. The pathways enriched by genes in trophoblasts, smooth muscle cells (SMC), and immune cells of PAS were mainly associated with immune and inflammation. We identified elevated expression of the angiogenesis-stimulating gene PTK2, alongside the cell proliferation-enhancing gene EGFR, within the trophoblasts of PAS group. Trophoblasts mainly contributed the enhancement of HLA-G and EBI3 signaling, which is crucial in establishing immune escape. Meanwhile, SMC regions in PAS exhibited upregulation of immunomodulatory markers such as CD274, HAVCR2, and IDO1, with CD274 expression experimentally verified to be increased in the invasive SMC areas of the PAS group.ConclusionsThis study provided information of cellular composition and spatial organization in PAS at single-cell and spatial level. The dysregulated expression of genes in PAS revealed a complex interplay between enhanced immune escape in trophoblasts and immune tolerance in SMCs during invasion in PAS. These findings will enhance our understanding of PAS pathogenesis for developing potential therapeutic strategies.

Altered extracellular matrix–related pathways accelerate the transition from normal to prefibroid myometrium in Black women

BackgroundBlack women experience a disproportionate impact of uterine fibroids compared to White women, including earlier diagnosis, higher frequency, and more severe symptoms. The etiology underlying this racial disparity remains elusive. ObjectiveThe aim of this study was to evaluate the molecular differences in normal myometrium (fibroid-free uteri) and at-risk myometrium (fibroid-containing uteri) tissues in Black and White women. Study designWe conducted whole-genome RNA-seq on normal and at-risk myometrium tissues obtained from both self-identified Black and White women (not Hispanic or Latino) to determine global gene expression profiles and to conduct enriched pathway analyses (n=3 per group). We initially assessed the differences within the same type of tissue (normal or at-risk myometrium) between races. Subsequently, we analyzed the transcriptome of normal myometrium compared to at-risk myometrium in each race and determined the differences between them. We validated our findings through real-time PCR (sample size range=5-12), western blot (sample size range=5-6), and immunohistochemistry techniques (sample size range=9-16). ResultsThe transcriptomic analysis revealed distinct profiles between Black and White women in normal and at-risk myometrium tissues. Interestingly, genes and pathways related to extracellular matrix and mechanosensing were more enriched in normal myometrium from Black than White women. Transcription factor enrichment analysis detected greater activity of the serum response transcription factor positional motif in normal myometrium from Black compared to White women. Furthermore, we observed increased expression levels of myocardin-related transcription factor-serum response factor and the serum response factor in the same comparison. In addition, we noted increased expression of both mRNA and protein levels of vinculin, a target gene of the serum response factor, in normal myometrium tissues from Black women as compared to White women. Importantly, the transcriptomic profile of normal to at-risk myometrium conversion differs between Black and White women. Specifically, we observed that extracellular matrix-related pathways are involved in the transition from normal to at-risk myometrium and that these processes are exacerbated in Black women. We found increased levels of Tenascin C, type I collagen alpha 1 chain, fibronectin and, phospho-p38 MAPK (Thr180/Tyr182, active) protein levels in at-risk over normal myometrium tissues from Black women, whereas such differences were not observed in samples from White women. ConclusionsThese findings indicate that the racial disparities in uterine fibroids may be attributed to heightened production of extracellular matrix in the myometrium in Black women, even before the tumors appear. Future research is needed to understand early life determinants of the observed racial differences.

Estrogen, estrogen receptor and miR-21 in adenomyosis: their pathogenic roles and regulatory interactions

To explore the pathogenic roles of miR-21, estrogen (E2), and estrogen receptor (ER) in adenomyosis. We examined the expression levels of miR-21 in specimens of adenomyotic tissue and benign cervical lesions using qRT-PCR. In primary cultures of cells isolated from the adenomyosis lesions, the effect of ICI82780 (an ER inhibitor) on miR-21 expression levels prior to E2 activation or after E2 deprivation were examined with qRT-PCR. We further assessed the effects of a miR-21 mimic or an inhibitor on proliferation, apoptosis, migration and autophagy of the cells. The expression level of miR-21 was significantly higher in adenomyosis tissues than in normal myometrium (P < 0.05). In the cells isolated from adenomyosis lesions, miR-21 expression level was significantly higher in E2 activation group than in ER inhibition + E2 activation group and the control group (P < 0.05); miR-21 expression level was significantly lower in cells in E2 deprivation+ER inhibition group than in E2 deprivation group and the control group (P < 0.05). The adenomyosis cells transfected with miR-21 inhibitor showed inhibited proliferation and migration, expansion of mitochondrial endoplasmic reticulum, increased lysosomes, presence of autophagosomes, and increased cell apoptosis, while transfection of the cells with the miR-21 mimic produced the opposite effects. MiR-21 plays an important role in promoting proliferation, migration, and antiapoptosis in adenomyosis cells by altering the cell ultrastructure, which may contribute to early pathogenesis of the disease. In addition to binding with E2, ER can also regulate miR-21 through other pathways to participate in the pathogenesis of adenomyosis, thus having a stronger regulatory effect on miR-21 than E2.

Uterine smooth muscle tumors: a multicenter, retrospective, comparative study of clinical and ultrasound features

ObjectiveTo evaluate a wide range of clinical and ultrasound characteristics of different uterine smooth muscle tumors to identify features capable of discriminating between these types.MethodsThis was a retrospective, multicenter study...

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).

Can an apparent diffusion coefficient of uterine fibroid before uterine artery embolization predict potential fibroid response?

ACOG guidance confirms the use of uterine artery embolisation (UAE) as an alternative to hysterectomy or myomectomy. The main objective of this article is to evaluate the ability of preoperative magnetic resonance ımaging (MRI) to study the relationship between uterine fibroid reduction and diffusion coefficient (ADC) value after UAE. This is a relevant topic with the growing interest in using ADC as a noninvasive imaging biomarker for monitoring tissue changes and predicting uterine fibroid response to UAE over the past years. In this prospective controlled non-randomized trial; uterine fibroid volume, fibroid diameter, uterine volume, fibroid ADC and normal myometrium ADC were recorded before and after UAE. Wilcoxon test was used in the analysis of the dependent quantitative data. Pearson correlation coefficients were calculated between post-UAE uterine volume, fibroid volume, and average fibroid diameter reduction and the patient's age, parity, gravidity, fibroid ADC and myometrial ADC before UAE. The mean fibroid volume reduction was 36.0% (range between 17.3-77.7%). Mean fibroid diameter, fibroid volume, uterine volume, and myometrium ADC values after UAE were significantly lower than before the procedure (p = 0.002, < 0.001, 0.001, 0.006 respectively), but the decrease in fibroid ADC is not significant. As a result decrease in fibroid volume was greater as pre-UAE fibroid ADC values increased, and that finding may contribute to the selection of the patients for the procedure. The ADC value before UAE was positively correlated with fibroid volume reduction.

O-265 Increased CD8+ T-cell exhaustion due to high NKG2A/HLA-E engagement and IL-15 induction in adenomyosis

Abstract Study question Is there dysregulation of immune cells in the local uterine microenvironment of patients with adenomyosis? If any, what’s the underlying mediators and mechanisms? Summary answer Increased CD8+ T-cell exhaustion due to enhanced HLA-E/NKG2A engagement and IL-15 induction in ectopic adenomyotic lesions existed in human patients and mice models with adenomyosis. What is known already The immunological dysfunction has long been considered in the pathogenesis of adenomyosis. However, high heterogeneity in immune cell alterations and contradictory results were reported due to the technique limitation of immunohistochemistry mainly used in previous studies. A comprehensive picture of the dysregulation of immune cells in the local uterine microenvironment of patients with adenomyosis is still lacking. Specially, increased number of CD8+ T cells have been reported in adenomyotic patients, whether functional deficiency of CD8+ T cells may contribute to the abnormal accumulation and residency of ectopic endometrial cells at pathological sites remains unknown. Study design, size, duration The paired ectopic lesions and eutopic endometrium from 80 patients with adenomyosis, and normal endometrium and myometrium from 62 patients with uterine myoma (controls) were collected between 2021 to 2022. Mice models with adenomyosis by neonatal tamoxifen treatment were established for experiments in 2022. Participants/materials, setting, methods The alteration of immune subsets, NKG2A expression, exhausted phenotypes and CD8+ T-cell dysfunction by flow cytometry, the expression/localization of HLA-E and IL-15 by immunohistochemistry were investigated in uterine tissues of patients and mice models with adenomyosis. CD8+ T cells were sorted, and in-vitro cultured with recombinant human transforming growth factor-β (rhTGF-β) or rhIL-15 to determine the NKG2A expression. Correlation analyses were conducted for association between immune disturbances and disease severity (dysmenorrhea, local/diffuse lesion, CA125 level). Main results and the role of chance We found that an increase in CD8+ T cell number was the predominant alteration in ectopic lesions in patients with adenomyosis, and it was significantly associated with the severity of adenomyosis. For CD8+ T cells in adenomyotic lesions, the expression of NKG2A was positively correlated with CD94 expression, and CD94/NKG2A mainly enriched on CD103+ tissue resident CD8+ T subsets. The NKG2A+CD8+ T cells showed exhausted phenotypes characterized by co-expression of multiple immune inhibitory molecules (PD1, LAG3, TIM3, and TOX) and decreased degranulation capability demonstrated by reduced expression of perforin, granzyme B, and CD107. Importantly, the exhausted NKG2A+CD8+ T-cell subset was associated with disease severity and was significantly increased in ectopic uterine tissues of human patients and mice models with adenomyosis. The expression of NKG2A on CD8+ T cells was markedly upregulated by exogenous rhIL-15 but not rhTGF-β treatment. Additionally, we also found increased co-expression of IL-15 and the NKG2A ligand HLA-E on the glandular epithelial cells, where the NKG2A+CD8+ T cells were closely distributed around the gland, in ectopic adenomyotic microenvironment of patients and mice models with adenomyosis. Limitations, reasons for caution The control uterine samples were collected from patients with uterine myoma. Although they were pathologically confirmed as normal, the potential effects are unknown and cannot be excluded. Further studies in larger independent populations and exploration on the functional significance of CD8+ T-cell exhaustion in adenomyosis pathogenesis is warranted. Wider implications of the findings Our study reveals a previously unrecognized role for CD8+ T-cell exhaustion in the pathogenesis of adenomyosis and suggests that therapeutic interventions targeting and reinvigorating exhausted CD8+ T cells by NKG2A blockade may be beneficial for patients with adenomyosis. Trial registration number not applicable

Transabdominal 2-Dimensional Shear-Wave Elastography for Diagnostic Differentiation Between Adenomyosis Leiomyoma and Normal Myometrium

Transabdominal 2-Dimensional Shear-Wave Elastography for Diagnostic Differentiation Between Adenomyosis Leiomyoma and Normal Myometrium

MMP14 expression and collagen remodelling support uterine leiomyosarcoma aggressiveness.

Fibrillar collagen deposition, stiffness and downstream signalling support the development of leiomyomas (LMs), common benign mesenchymal tumours of the uterus, and are associated with aggressiveness in multiple carcinomas. Compared with epithelial carcinomas, however, the impact of fibrillar collagens on malignant mesenchymal tumours, including uterine leiomyosarcoma (uLMS), remains elusive. In this study, we analyse the network morphology and density of fibrillar collagens combined with the gene expression within uLMS, LM and normal myometrium (MM). We find that, in contrast to LM, uLMS tumours present low collagen density and increased expression of collagen-remodelling genes, features associated with tumour aggressiveness. Using collagen-based 3D matrices, we show that matrix metalloproteinase-14 (MMP14), a central protein with collagen-remodelling functions that is particularly overexpressed in uLMS, supports uLMS cell proliferation. In addition, we find that, unlike MM and LM cells, uLMS proliferation and migration are less sensitive to changes in collagen substrate stiffness. We demonstrate that uLMS cell growth in low-stiffness substrates is sustained by an enhanced basal yes-associated protein 1 (YAP) activity. Altogether, our results indicate that uLMS cells acquire increased collagen remodelling capabilities and are adapted to grow and migrate in low collagen and soft microenvironments. These results further suggest that matrix remodelling and YAP are potential therapeutic targets for this deadly disease.

Prevalence and clinical significance of co-existing mutations in MED12 and FH in uterine fibroids of Australian women.

Uterine fibroids are exceedingly common benign tumours of the female reproductive system and cause severe symptoms, including acute pain, bleeding, and infertility. Fibroids are frequently associated with genetic alterations affecting mediator complex subunit 12 (MED12), fumarate hydratase (FH), high mobility group AT-hook 2 (HMGA2) and collagen, type IV alpha 5 and alpha 6 (COL4A5-COL4A6). Recently, we reported MED12 exon 2 mutations in 39 out of 65 uterine fibroids (60%) from 14 Australian patients. The aim of this study was to evaluate the status of FH mutations in MED12 mutation-positive and mutation-negative uterine fibroids. FH mutation screening of altogether 65 uterine fibroids and corresponding adjacent normal myometrium (n = 14) was carried out by Sanger sequencing. Three out of 14 patients displayed somatic mutations in FH exon 1 in addition to harbouring MED12 mutation in uterine fibroids. This study is the first to report that the mutations in MED12 and FH co-exist in uterine fibroids of Australian women.

Loss of dystrophin is common in uterine leiomyosarcoma: a potential biomarker for clinical application.

Uterine leiomyosarcoma (LMS) is a deadly disease with high rates of recurrence and a poor prognosis. Its tumorigenesis remains largely unknown, and no specific biomarkers can be used for the differential diagnosis of LMS from other mimics. Recent whole-genome studies revealed a loss of dystrophin is common in LMS, especially in uterine LMS. To investigate the expression pattern of dystrophin expression across different types of uterine smooth muscle tumors, immunohistochemistry was performed, including usual-type leiomyoma, fumarate hydratase-deficient leiomyoma, leiomyoma with bizarre nuclei, conventional LMS, and normal myometrium for this study. To further evaluate the genomic change in dystrophin gene region, whole-genome sequencing in 10 LMS cases were analyzed. Dystrophin expression was detected in 94% (45/48) of myometrium, 97% (34/35) of usual-type leiomyoma, 84% (26/31) of fumarate hydratase-deficient leiomyoma, 60% (12/20) of leiomyoma with bizarre nuclei, and 18% (6/34) of LMS. Loss of dystrophin expression was significantly different between benign and malignant tumors (LMS cases counted as malignant only) (p<0.01). Of note, copy number loss in the dystrophin genomic region was found in all 10 cases of LMS. Additionally, patients with dystrophin-positive LMS tend to have a better overall survival than patients with dystrophin-negative LMS.

High levels of hypusinated eIF5A in leiomyoma and leiomyosarcoma pathologies: a possible novel therapeutic target

Research questionIs the hypusinated form of the eukaryotic translation initiation factor 5A (EIF5A) present in human myometrium, leiomyoma and leiomyosarcoma, and does it regulate cell proliferation and fibrosis? DesignThe hypusination status of eIF5A in myometrial and leiomyoma patient-matched tissues was evaluated by immunohistochemistry and Western blotting as well as in leiomyosarcoma tissues by immunohistochemistry. Myometrial, leiomyoma and leiomyosarcoma cell lines were treated with N1-guanyl-1,7-diaminoheptane (GC-7), responsible for the inhibition of the first step of eIF5A hypunization, and the proliferation rate was determined by MTT assay; fibronectin expression was analysed by Western blotting. Finally, expression of fibronectin in leiomyosarcoma tissues was detected by immunohistochemistry. ResultsThe hypusinated form of eIF5A was present in all tissues examined, with an increasing trend of hypusinated eIF5A levels from normal myometrium to neoplastic benign leiomyoma up to neoplastic malignant leiomyosarcoma. The higher levels in leiomyoma compared with myometrium were confirmed by Western blotting (P = 0.0046). The inhibition of eIF5A hypusination, with GC-7 treatment at 100 nM, reduced the cell proliferation in myometrium (P = 0.0429), leiomyoma (P = 0.0030) and leiomyosarcoma (P = 0.0044) cell lines and reduced the expression of fibronectin in leiomyoma (P = 0.0077) and leiomyosarcoma (P = 0.0280) cells. The immunohistochemical staining of leiomyosarcoma tissue revealed that fibronectin was highly expressed in the malignant aggressive (central) part of the leiomyosarcoma lesion, where hypusinated eIF5A was also highly represented. ConclusionsThese data support the hypothesis that eIF5A may be involved in the pathogenesis of myometrial benign and malignant pathologies.

Management of interstitial pregnancy using laparoscopic salpingectomy with complete removal of the interstitial portion

To perform laparoscopic salpingectomy, including the entire interstitial portion of the fallopian tube, in the management of interstitial pregnancy. A step-by-step explanation of the surgical procedure using video with narration. Obstetrics and Gynecology department of a hospital. A 23-year-old woman, gravida 1 para 0, presented asymptomatically to our hospital to undergo a pregnancy test. Her last menstrual period had occurred 6 weeks previously. Transvaginal ultrasound showed an empty uterine cavity and a right interstitial mass of 3.2 × 2.6 × 2.5 cm. It contained a chorionic sac and an embryonic bud of 0.2 cm long with a heartbeat and the presence of an "interstitial line sign." The myometrial layer surrounding the chorionic sac was 1 mm. The patient's beta-human chorionic gonadotropin level was 10,123 mIU/mL. On the basis of the anatomy of the interstitial portion of the fallopian tube, we treated interstitial pregnancy using laparoscopic salpingectomy, with complete removal of the interstitial portion containing the product of conception. The interstitial fallopian tube originates at the tubal ostium and follows a tortuous intramural course, progressing laterally away from the uterine cavity toward the isthmic portion. It is lined by muscular layers and an inner epithelium layer. The main blood supply of the interstitial portion is from the uterine artery's ascending branches to the fundus, extending a branch that supplies the cornu and the interstitial portion. Our approach has 3 key steps: 1) dissecting and coagulating the branch extending from the ascending branches to the fundus of the uterine artery, 2) incising the cornual serosa at the junction of the purple-blue interstitial pregnancy and the normal color myometrium, and 3) resecting the interstitial portion containing the product of conception along the outer layer of the oviduct without rupture. The interstitial portion containing the product of conception was removed entirely along the outer layer of the fallopian tube as a natural capsule without rupture. The surgery lasted for 43 min, and the volume of intraoperative blood loss was 5 mL. The pathology was confirmatory for interstitial pregnancy. The patient's beta-human chorionic gonadotropin levels decreased optimally. She had a normal postoperative course. This approach reduces intraoperative blood loss, minimizes myometrial loss and thermal injury, and effectively avoids persistent interstitial ectopic pregnancy. It is not limited by the device used, does not increase the surgery cost, and is greatly useful in treating a selected nonruptured distally or centrally implanted interstitial pregnancy.