Development Of Leiomyomas Research Articles

The Functional Role of the Long Non-Coding RNA LINCMD1 in Leiomyoma Pathogenesis

Existing evidence indicates that LINCMD1 regulates muscle differentiation-related gene expression in skeletal muscle by acting as a miRNA sponge, though its role in leiomyoma development is still unknown. This study investigated LINCMD1′s involvement in leiomyoma by analyzing paired myometrium and leiomyoma tissue samples (n = 34) from patients who had not received hormonal treatments for at least three months prior to surgery. Myometrium smooth muscle cells (MSMCs) were isolated, and gene expression of LINCMD1 and miR-135b was assessed via qRT-PCR, while luciferase assays determined the interaction between LINCMD1 and miR-135b. To examine the effects of LINCMD1 knockdown, siRNA transfection was applied to a 3D MSMC spheroid culture, followed by qRT-PCR and Western blot analyses of miR-135b, APC, β-Catenin and COL1A1 expression. The results showed that leiomyoma tissues had significantly reduced LINCMD1 mRNA levels, regardless of patient race or MED12 mutation status, while miR-135b levels were elevated compared to matched myometrium samples. Luciferase assays confirmed LINCMD1′s role as a sponge for miR-135b. LINCMD1 knockdown in MSMC spheroids increased miR-135b levels, reduced APC expression, and led to β-Catenin accumulation and higher COL1A1 expression. These findings highlight LINCMD1 as a potential therapeutic target to modulate aberrant Wnt/β-Catenin signaling in leiomyoma.

Bleeding vs Thrombosis: Treatment Strategy for Women Having Large Uterine Fibroids and DVT

Large uterine fibroids (UFs) are clonal neoplasms of the uterus that form during the mid and late-aged women. Such women generally consume oral contraceptive pills, tranexamic acid, or NSAIDS to manage heavy menstrual bleeding (HMB) and associated complications while waiting for a conclusive procedure or avoiding hysterectomy. The procoagulant effect of these medicinal agents can result in venous stasis of the lower limbs, leading to deep vein thrombosis (DVT), a challenging complication with HMB. We examine the complicated state of heavy bleeding with thrombosis and explore better management options. It has been seen that women with hypothyroidism have an increased risk of getting DVT due to an alteration in the coagulation system. These incidences are mostly associated with higher uterine weight, which is related to the extrinsic compression of the inferior vena cava. In such cases, the occurrence of postoperative thrombosis is riskier if hysterectomy/myomectomy is the only option. Utilization of appropriate anticoagulants with modification of the steroid-hormone system using hormone agonists or antagonists (e.g., levonorgestrel intrauterine system, high-dose progestin-only therapy, danazol, aromatase inhibitors, Vitamin-D supplements or selective estrogen receptor modulators) could be an effective technique, but adverse consequences of continued use should be monitored. More research is needed into the basic biology associated with the role of growth factors and genetic alterations in these malignancies. The development of new leiomyomas following conservative therapy is also a significant issue.

Periostin's role in uterine leiomyoma development: a mini-review on the potential periostin poses as a pharmacological intervention for uterine leiomyoma.

Uterine leiomyomas, also known as fibroids or myomas, occur in an estimated 70-80% of reproductive aged women. Many experience debilitating symptoms including pelvic pain, abnormal uterine bleeding (AUB), dyspareunia, dysmenorrhea, and infertility. Current treatment options are limited in preserving fertility, with many opting for sterilizing hysterectomy as a form of treatment. Currently, surgical interventions include hysterectomy, myomectomy, and uterine artery embolization in addition to endometrial ablation to control AUB. Non-surgical hormonal interventions, including GnRH agonists, are connotated with negative side effects and are unacceptable for women desiring fertility. Periostin, a regulatory extra cellular matrix (ECM) protein, has been found to be expressed in various gynecological diseases including leiomyomas. We previously determined that periostin over-expression in immortalized myometrial cells led to the development of a leiomyoma-like cellular phenotype. Periostin is induced by TGF-β, signals through the PI3K/AKT pathway, induces collagen production, and mediates wound repair and fibrosis, all of which are implicated in leiomyoma pathology. Periostin has been linked to other gynecological diseases including ovarian cancer and endometriosis and is being investigated as pharmacological target for treating ovarian cancer, post-surgical scarring, and numerous other fibrotic conditions. In this review, we provide discussion linking pathological inflammation and wound repair, with a TGF-β-periostin-collagen signaling in the pathogenesis of leiomyomas, and ultimately the potential of periostin as a druggable target to treat leiomyomas.

Leiomyoma and the importance of genetic variation on genes related to the vasculature system - CβS, MTHFR, NOS3, CYBA, and ACE1

ObjectiveThe link between the systemic vasculature system and tumor biology is here investigated by studying the contribution of CβS (844ins68), MTHFR (677C > T), NOS3 (4a/4b), CYBA (C242T), and ACE1 (I/D) genes to leiomyoma onset, uterus and leiomyoma volumes. MethodsDNA samples from 130 women with leiomyomas and 527 from healthy women were genotyped by PCR or PCR-RFLP. Qui-square (χ2) or Fisher’s exact test were used to test associations. All the mentioned tests were performed in IBM® SPSS® Statistics Version 28. Statistical significance was defined as a p-value < 0.05. ResultsResults revealed that CβS (in the codominant and allelic models, p = 0.044 and, p = 0.015, OR = 1.791 [1.114–2.879], respectively), MTHFR (in the codominant, allelic and dominant models, p = 0.009, p = 0.002, OR = 0.585 [0.416–0.824] and p = 0.003, OR = 0.527 [0.346–0.802], respectively) and ACE1 (dominant model, p = 0.045, OR = 0.639 [0.411–0.992]) genes are associated with leiomyoma onset. NOS3 4a4a genotype is associated with a lower uterus volume (p = 0.004). This study also uncovers intriguing epistatic interactions among some genes that further accentuate their roles in disease modulation. Indeed, the epistatic interactions between the CC genotype (MTHFR) and (+/+) (CβS; p = 0.003), 4b4b (NOS3; p = 0.006, OR = 2.050 [1.223–3.439]) or DD (ACE1; p < 0.001, OR = 2.362 [1.438–3.880]) were shown to be associated with the disease, while 4a presence (NOS3) in epistasis with I presence (ACE1), increased the effect protection having just the I allele presence (p = 0.029, OR = 0.446 [0.214–0.930]). ConclusionsWe conclude that variation in genes related to the systemic vascular system can play a role in the onset and development of leiomyoma.

Vitamin D and its binding protein in patients with leiomyomas.

This study examined the levels of VitD, VitD binding protein (DBP), and free VitD in leiomyomas patients and their association with the quantity, dimensions, and site of fibroid growths. Additionally, we evaluated the potentiality of employing these factors as a biomarker tool for the diagnosis and assessment of uterine fibroid progression. This study involved the participation of 55 women with leiomyomas and 50 healthy women. We utilized commercial ELISA kits to measure the levels of total VitD and DBP in their serum. Additionally, we calculated the levels of free VitD and the ratio of VitD to DBP. Moreover, we determined the number, size, and location of the leiomyomas in the patients. There were no significant differences in the levels of total VitD between the groups. However, patients had significantly lower levels of free VitD and higher levels of DBP compared to the control group. The size of the largest leiomyomas showed a negative relationship with free VitD and a positive relationship with DBP. Receiver operating characteristic analyses, showed that the cut-off value for free VitD was 4.47 pg/mL, with a sensitivity of 75.6% and a specificity of 74.4%. The cut-off value for DBP was 256.2 μg/mL, with a sensitivity of 86% and a specificity of 70.3%. Free VitD and DBP potentially contribute to the development of leiomyomas and are linked to the size of these tumors. The measurement of serum levels of these factors could serve as additional biomarkers for the diagnosis of leiomyomas.

Causal relationship between genetically predicted uterine leiomyoma and cancer risk: a two-sample Mendelian randomization.

Studies have demonstrated that hormonal imbalance, such as elevated level of estrogen or reduced level of progesterone, was the main inducing factor of uterine leiomyoma (UL) development and some cancers. UL has been reported to be associated with several cancers in observational studies. However, the causal associations between UL and cancers remain unclear. A two-sample Mendelian randomization (MR) analysis was conducted to investigate the causal associations between UL and 16 site-specific cancers using the public databases. Four methods, namely, the inverse variance weighting (IVW), MR-Egger, weighted median, and weighted mode, were applied in our MR analysis. Sensitivity tests were also performed to evaluate the robustness of these causal associations. The IVW analysis indicated that genetically predicted UL increased the risk of low malignant potential ovarian cancer [odds ratio (OR) = 1.22, 95% confidence interval (CI): 1.06-1.40, p = 0.004], serous ovarian cancer (OR = 1.29, 95% CI: 1.10-1.52, p = 0.002), invasive mucinous ovarian cancer (OR = 1.24, 95% CI: 1.08-1.44, p = 0.003), clear cell ovarian cancer (OR = 1.25, 95% CI: 1.03-1.51, p = 0.023), breast cancer (OR = 1.07, 95% CI: 1.02-1.11, p = 0.002), and brain tumor (OR = 1.23, 95% CI: 1.06-1.42, p = 0.007). Conversely, genetically predicted UL reduced the risk of gastric cancer (OR = 0.91, 95% CI: 0.85-0.98, p = 0.008). The causal effects were consistent in the sensitivity analysis. Our results demonstrated that UL exhibits a causal relationship with high risk of several cancers. We suggest reinforcing the cancer screening in UL patients to enable the early detection of cancers.

Cryptic splice mutation in the fumarate hydratase gene in patients with clinical manifestations of Hereditary Leiomyomatosis and Renal Cell Cancer.

Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is an autosomal dominant condition characterized by the development of cutaneous and uterine leiomyomas and risk for development of an aggressive form of papillary renal cell cancer. HLRCC is caused by germline inactivating pathogenic variants in the fumarate hydratase (FH) gene, which encodes the enzyme that catalyzes the interconversion of fumarate and L-malate. We utilized enzyme and protein mobility assays to evaluate the FH enzyme in a cohort of patients who showed clinical manifestations of HLRCC but were negative for known pathogenic FH gene variants. FH enzyme activity and protein levels were decreased by 50% or greater in three family members, despite normal FH mRNA expression levels as measured by quantitative PCR. Direct Nanopore RNA sequencing demonstrated 57 base pairs of retained intron sequence between exons 9 and 10 of polyadenylated FH mRNA in these patients, resulting in a truncated FH protein. Genomic sequencing revealed a heterozygous intronic alteration of the FH gene (chr1: 241498239T/C) resulting in formation of a splice acceptor site near a polypyrimidine tract, and a uterine fibroid obtained from a patient showed loss of heterozygosity at this site. The same intronic FH variant was identified in an unrelated patient who also showed a clinical phenotype of HLRCC. These data demonstrate that careful clinical assessment as well as biochemical characterization of FH enzyme activity, protein expression, direct RNA sequencing, and genomic DNA sequencing of patient-derived cells can identify pathogenic variants outside of the protein coding regions of the FH gene.

In deep bioinformatic characterization of a novel fumarate hydratase variant FH c.199T > G; (p.Tyr67Asp) in hereditary leiomyomatosis and renal cell carcinoma.

Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is a rare, autosomal dominant tumor predisposition syndrome characterized by variable development of multiple skin and uterus leiomyomas and aggressive forms of renal cell carcinoma (RCC). Mutations in fumarate hydratase (FH), one of the proteins in homologous recombination repair, precede the development of HLRCC with high penetrance. Considering the risk of early metastasis of RCC, FH has been included in mutation screening panels. The identification of a pathogenic FH variant guides the screening for tumors in the carriers. However, variants of uncertain significance (VUS) are frequent findings, limiting the clinical value of the mutation screening. Here, we describe the associated phenotype and an in-depth, multi-step Bioinformatic evaluation of the germline FH c.199T > G (p.Tyr67 > Asp) variant segregated in an HLRCC family. Evidence for FH c.199T > G; (p.Tyr67Asp) pathogenicity includes the variant segregation with the disease in three affected family members, its absence in populational databases, and the deep evolutionary conservation of the Tyr67 residue. At the protein level, this residue substitution causes the loss of molecular bonds and ionic interactions, affecting molecular dynamics and protein stability. Considering ACMG/AMP criteria, we propose the reclassification of the FH c.199T > G; (p.Tyr67Asp) variant to "likely pathogenic". In addition, the in-depth, in silico approach used here allowed us to understand how and why FH c.199T > G; (p.Tyr67Asp) could cause HLRCC. This could help in clinical management decisions concerning the monitoring of unaffected family members having this variant.

Beta blockers reduce uterine fibroid incidence in hypertensive women

ObjectiveIs prior beta blocker (BB) use associated with reduced odds of the clinical incidence of leiomyomas? What is known alreadyIn-vitro and in-vivo evidence has supported the role of beta receptor blockade in reducing leiomyoma cell proliferation and growth. However, no population-based study to date has investigated this potential association. Study design, size, durationA nested case-control study was conducted in a population of women aged 18–65 with arterial hypertension (n = 699,966). Cases (n = 18,918) with a leiomyoma diagnosis were matched to controls (n = 681,048) with no such diagnosis at a 1:36 ratio by age and region of origin within the United States. Participants/materials, setting, methodsThis population was assembled from the Truven Health MarketScan® Research Database, which includes health insurance claims from January 1st, 2012 to December 31st, 2017. Prior use of BB wasdetermined fromoutpatient drug claims and leiomyoma development was indicated by a first-time diagnosis code. We conducted a conditional logistic regression to determine the odds of uterine fibroid development in women with prior use of BB compared to women with no such history. We then conducted subset analyses, stratifying the women by age group and by type of BB. ResultsWomen on a BB experienced 15% reduced odds of developing clinically recognized leiomyoma compared to non-users (OR 0.85, 95% CI 0.76–0.94). This association was significant for the 30–39 age group (OR 0.61, 95% CI 0.40–0.93) but no other age group. Of the BBs, propranolol (OR 0.58, 95% CI 0.36–95) demonstrated a significant association with reduced leiomyoma incidence and metoprolol (OR 0.82, 95% CI 0.70–0.97) was associated with lower uterine fibroid incidence after adjustment for comorbidities. ConclusionsHypertensive women with prior BB use experienced reduced odds of developing clinically recognized leiomyoma compared to non-users. A key predisposing risk factor for uterine leiomyoma is elevated blood pressure. Thus, the results of this analysis may have clinical relevance to women with hypertension, as the use of this drug may introduce a dual benefit of managing hypertension as well as curbing an increased risk of leiomyomas.

Correlation between inflammatory marker and lipid metabolism in patients with uterine leiomyomas.

Obesity is a risk factor for the development of uterine leiomyoma (UL), and the inflammatory response plays a key role in the pathogenesis of UL. Our objective was to assess whether there was an independent relationship between inflammatory markers and triglycerides (TG) in patients with UL. 1,477 UL participants who were hospitalized at the Jining Medical University between January 2016 and December 2022 were included in this cross-sectional study. The independent and dependent variables measured at baseline were inflammatory markers and TG levels, respectively. The covariates were age, body mass index (BMI), UL and menstrual status. Based on the number of fibroids, the study population was divided into Single-group and Multiple-group. Univariate and multiple regression analyses and stratified analyses revealed significant positive correlations between neutrophil-lymphocyte ratio and systemic immune inflammation index and TG, and significant negative correlations between monocyte-lymphocyte ratio and TG. The findings show a significant correlation between the inflammatory response and lipid metabolism levels in UL patients. This provides direction for further research into the pathophysiology of UL and also helps to formulate hypotheses for predictive models of UL.

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.

Adipocyte and Adipokines Promote a Uterine Leiomyoma Friendly Microenvironment.

Uterine leiomyomas are the most common benign tumors of the female reproductive system. Obese individuals have a higher burden of uterine leiomyoma, yet the mechanism relating obesity and leiomyoma development remains unknown. In this study, we observe the effect of adipocyte coculture and leptin treatment on human myometrium and leiomyoma cells. We isolated primary leiomyoma and myometrium cells from hysterectomy or myomectomy patients. Protein expression levels of phosphorylated ERK1/2/total ERK1/2, phosphorylated STAT3/total STAT3, and phosphorylated AKT1/2/3/total AKT1/2/3 were quantified using immunoblotting in immortalized and primary leiomyoma and myometrial cells cocultured with human adipocytes and treated with leptin. An enzyme-linked immunosorbent assay (ELISA) was used to assess pro-inflammatory, fibrotic, and angiogenic factors in immortalized human myometrium and leiomyoma cells treated with leptin. The effects of STAT3, ERK, and AKT inhibitors were assessed in leiomyoma cell lines additionally cultured with adipocytes. Adipocyte coculture and leptin treatment increases the expression of JAK2/STAT3, MAPK/ERK, and PI3K/AKT signaling while inhibitors suppressed this effect. Leptin induces a tumor-friendly microenvironment through upregulation of pro-inflammatory (IFNγ, IL-8, IL-6, GM-CSF, MCP-1, and TNF-α), fibrotic (TGF-β1, TGF-β2, and TGF-β3), and angiogenic (VEGF-A, HGF, and Follistatin) factors in human leiomyoma cells. Furthermore, adipocyte coculture and leptin treatment increases leiomyoma cells growth through activation of MAPK/ERK, JAK2/STAT3, and PI3k/AKT signaling pathways. Finally, STAT3, ERK, and AKT inhibitor treatment suppressed PCNA, TNF-α, TGF-β3, and VEGF-A intracellular staining intensity in both adipocyte coculture and leptin treated leiomyoma cells. These findings suggest that, in obese women, adipocyte secreted hormone or adipocytes may contribute to leiomyoma development and growth by activating leptin receptor signaling pathways.

Integrating Spatial Transcriptomics and Single-nucleus RNA Sequencing Reveals the Potential Therapeutic Strategies for Uterine Leiomyoma.

Uterine leiomyoma is the most common gynecological tumor in reproductive women. Tumor-host interface is a complex ecosystem with intimate cell-cell communications and a critical scenario for tumor pathogenesis and progression. The pseudocapsule is the main tumor-host interface of uterine leiomyoma, but its cellular spatial disposition and gene expression are poorly explored. This study mapped the cellular architecture and corresponding gene profiles of the leiomyoma and its surrounding pseudocapsule by integrating spatial transcriptomics and single-nucleus RNA-sequencing at the first time. Here, we reported that estrogen receptor alpha and progesterone receptor mediated the occurrence and development of uterine leiomyoma and that estrogen receptor beta involved in the angiogenesis, which explained the effectiveness of hormonotherapy. Therapeutic targets including ERK1/ERK2 pathway and IGF1-IGF1R were found and might be applied for non-hormonal therapy of uterine leiomyoma. Furthermore, the injection of prostaglandin E2 was initially presented for bleeding control during myomectomy, injection site should be located at the junction between pseudocapsule and leiomyoma, and surrounding pseudocapsule should not be eliminated. Collectively, a single-cell and spatially resolved atlas of human uterine leiomyoma and its surrounding pseudocapsule was established. The results revealed potentially feasible strategies for hormonotherapy, non-hormonal targeted therapy and bleeding control during myomectomy.

Genetic Contribution of the Adrenergic, Cholinergic, and Serotonergic Systems to Leiomyoma Development and Treatment.

The link between the autonomic nervous system and tumor biology is being unfold. We aim to study the contribution of genes of the adrenergic (ADBR2 - rs1042713, NM_000024.6:c.46G>A, NP_000015.2:p. Gly16Arg), cholinergic (CHRNA5 - rs16969968, NM_000745.3:c.1192G>A, NP_000736.2:p.Asp398Asn), and serotonergic systems (SLC6A4 - 5-HTTVNTR-intron2, HTR2A - rs6313, NM_000621.5:c.102C>T, NP_ 001365853 .1: p. Ser 34=) to gynecological tumorigenesis and their treatment by embolization. A total of 517 DNA samples from women were analyzed. Samples were genotyped by PCR, PCR-RFLP and EndPoint genotyping. Results show a statistically significant association between the AA genotype of the ADBR2 gene and GG genotype of the CHRNA5 gene with leiomyoma (OR = 2.311; p = 0.003 and OR = 2.165; p = 0.001, respectively), and the epistatic interaction between genotypes increases the risk (OR = 2.458; p= 0.043). The GG genotype (CHRNA5) shows a lower reduction of the volume of the main leiomyoma after treatment (p=0.015). Combination of the genotypes 12/12-AA (SLC6A4 - ADBR2) increases the risk to leiomyoma (OR = 2.540, p= 0.030). TT genotype of HTR2A gene in combination with any of the two risk genotypes (of ADBR2 or CHRNA5) increases substantially the risk (OR = 5.266, p = 0.006; OR = 6.364, p=0.007, respectively). We conclude that ADBR2 and CHRNA5 genes have a relevant role that is enhanced by the epistatic relationship with the genes HTR2A and SLC6A4. CHRNA5 gene may also be a modulator of the success of embolization. We confirm the contribution of the genetics of Autonomous Nervous System to tumor biology.

Mono-(2-ethyl-5-hydroxyhexyl) phthalate promotes uterine leiomyoma cell survival through tryptophan-kynurenine-AHR pathway activation

Uterine leiomyoma is the most common tumor in women and causes severe morbidity in 15 to 30% of reproductive-age women. Epidemiological studies consistently indicate a correlation between leiomyoma development and exposure to endocrine-disrupting chemical phthalates, especially di-(2-ethylhexyl) phthalate (DEHP); however, the underlying mechanisms are unknown. Here, among the most commonly encountered phthalate metabolites, we found the strongest association between the urine levels of mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), the principal DEHP metabolite, and the risk of uterine leiomyoma diagnosis (n = 712 patients). The treatment of primary leiomyoma and smooth muscle cells (n = 29) with various mixtures of phthalate metabolites, at concentrations equivalent to those detected in urine samples, significantly increased cell viability and decreased apoptosis. MEHHP had the strongest effects on both cell viability and apoptosis. MEHHP increased cellular tryptophan and kynurenine levels strikingly and induced the expression of the tryptophan transporters SLC7A5 and SLC7A8, as well as, tryptophan 2,3-dioxygenase (TDO2), the key enzyme catalyzing the conversion of tryptophan to kynurenine that is the endogenous ligand of aryl hydrocarbon receptor (AHR). MEHHP stimulated nuclear localization of AHR and up-regulated the expression of CYP1A1 and CYP1B1, two prototype targets of AHR. siRNA knockdown or pharmacological inhibition of SLC7A5/SLC7A8, TDO2, or AHR abolished MEHHP-mediated effects on leiomyoma cell survival. These findings indicate that MEHHP promotes leiomyoma cell survival by activating the tryptophan-kynurenine-AHR pathway. This study pinpoints MEHHP exposure as a high-risk factor for leiomyoma growth, uncovers a mechanism by which exposure to environmental phthalate impacts leiomyoma pathogenesis, and may lead to the development of novel druggable targets.

Serum sestrin 2 levels in patients with uterine leiomyomas

Our aim was to evaluate SESN2 levels in patients with uterine leiomyomas by comparing serum SESN2 levels in myoma patients with the levels in healthy women to deepen our understanding of the pathophysiology of uterine leiomyomas. Patients 18–50 years of age who applied to the University of Health Sciences Turkey, Istanbul Kanuni Sultan Suleyman Training and Research Hospital between January and March 2021 and who were diagnosed with uterine leiomyoma were defined as the ‘myoma group’. The control group included patients without any sign of leiomyomas in routine ultrasonography. The patients’ demographic features, gynecological symptoms, myoma volume and classification were recorded. Serum SESN2 concentrations in venous blood samples were measured using a sandwich enzyme-linked immunosorbent assay (ELISA) kit. The study included 31 patients in the myoma group and 30 in the control group. The mean age/gravid/parity or BMI values did not differ significantly between the groups. The only gynecological symptom that showed a significant difference was menorrhagia. Serum SESN2 levels were significantly higher in the myoma group then the control groups (11.7 ± 2.5) (p < 0.001). In conclusion, although uterine leiomyoma is the most common benign tumour in women of reproductive age, there are no known markers for predicting the development of leiomyomas. Based on the results of the current study, SESN2 could be such a marker. IMPACT STATEMENT What is already known on this subject? Uterine leiomyoma is the most common type of benign tumour in women of reproductive age as well as the most common indication for a hysterectomy. Symptoms associated with uterine leiomyoma include abnormal bleeding, chronic pelvic pain, menorrhagia, dysmenorrhoea, dyspareunia and anaemia, which adversely affect the patient’s quality of life. Sestrins are a family of metabolic regulator proteins that play a potential role in carcinogenesis. What the results of this study add? This is the first study evaluating the role of sestrin in the development of uterine leiomyomas. Significantly higher levels of sestrin 2 (SESN2) were detected in patients with leiomyomas. What are the implications of these findings for clinical practice and/or further research? Although uterine leiomyoma is the most common type of benign tumour in women of reproductive age, there are still many unknowns regarding its pathophysiology. Further, there are still no known markers for predicting the development of leiomyomas. Hence, primary prevention is not possible. Based on the results of the current study SESN2, could be such a marker. Further studies are needed to confirm the results of this study.

In Silico Analysis of Plant-Based Bioactive Compounds Targeting Progesterone Receptors for the Treatment of Uterine Leiomyoma

Uterine leiomyomas are the most common benign gynaecological tumours that affect 50% - 60% of females belonging to reproductive age. The role of progesterone in the growth and development of leiomyomas is evident. Progesterone is regulated by progesterone receptors and it acts by activating PI3K/AKT pathway, by increasing proliferating cell nuclear antigen (PCNA) levels and Bcl-2 expression. Thus, modulating the receptor can help in the treatment of leiomyomas. The current study involves testing the efficiency of the various plant-based bioactive compounds to bind to the progesterone receptors, which may possess inhibitory or modulatory effects on or against progesterone receptors. These compounds can be further experimented with using other in vivo and in vitro methods to produce phytomedicine based on drugs for the treatment of uterine leiomyomas. This study is an in silico analysis performed using various bioinformatics databases and software. The receptor and ligand structures were retrieved from PDB and PubChem databases. Their pharmacokinetics and pharmacodynamics properties were analyzed using various databases. AutoDock4.2.6 was used for molecular docking, and the results were visualized using LigPlot+. Among 15 compounds selected, 8 compounds had higher binding energies than the control drug Ulipristal acetate i.e., -8.21Kcal/mol. Of which Ursolic acid, Vitamin-D2, and Betulinic acid had binding energies above -9.0 Kcal/mol. This concludes that these compounds can modulate the progesterone receptors, thus decreasing the proliferation and growth of uterine leiomyomas. Further studies can bring out the potential therapeutic effects of these bioactive compounds and they can act as a potential lead compound in the design of novel drug targets for the treatment of uterine leiomyoma.

STROMAL-VASCULAR PARAMETERS FOR PROLIFERATIVE LEIOMYOMAS OF THE BODY OF UTERUS IN WOMEN WITH REPRODUCTIVE PERIOD IN THE ASPECT OF AGE

Leiomyoma of the body of uterus is a tumor of unknown etiology, the frequency of which ranges from 5.4% to 77.0% and 80.0% of patients suffer from this problem during their lifetime. Despite the researches conducted up to date, the etiology of leiomyoma is still largely unknown and treatment strategy is limited due to the lack of information on the pathogenesis of this disease. Objectives and methods: 1. Ultrasonographic examination of leiomyoma nodes with color doppler mapping and resistance index detection. 2. Staining of preparations with hematoxylin and eosin to determine histostructure. 3. Staining of preparations with Masson Trichrome for detection of extracellular matrix. The percentage rate of fibrotic changes in the node was evaluated using the ImageJ program by processing the preparations stained with MASSON TRICHROME through a special algorithm. 4. Immunohistochemical evaluation of angiogenesis using CD34 marker. Morphometric analysis of blood vessels was performed with the special algorithm of CellProfiler program. The obtained results were processed by the computer program package of mathematical statistics - SPSS-21. Pearson x2 was used to check the confidence. Conclusions: 1. In the period before the nodes of uterine leiomyoma, a reshaped vascular collector is formed with sharp vascularization, which is a cover; and creates an ideal environment in the process of growth and development of leiomyoma; The growth of nodes up to 1 cm proceeds by diffusion from the adjacent muscle within the collector, as an independent autonomous unit, in conditions of scarcity of internal angiogenesis in the nodes (angiogenesis 45%, RI-0.48). 2. In nodes of proliferative leiomyoma with the size of up to 2 cm, there is an increase in internal (intranodal) vascularization, which confirms the active role of angiogenesis in the process of growth and development of leiomyoma; 3. Along with the growth of leiomyomas up to 3 cm, both in the muscle adjacent to the nodes (RI-0.61) and in the internal tissue of the nodes, there is a decrease in angiogenesis up to 25.6%, (RI-0.63); and in nodules up to 4 cm, a dramatic decrease in angiogenesis in the adjacent muscle (RI-0.53) and intranodal tissue up to 15.8% (RI-0.42). The mentioned fact confirms the complete exhaustion of the resources of remodeled blood vessels formed during the growth and development of leiomyoma within the collector by forming a hypoxic condition in the nodes. 4. In the proliferative, small growing leiomyomas, the proliferative activity of fibroblasts with the formation of extracellular matrix and the reproduction of fibrotic tissue is directly proportional to the age of the patient - with their decrease up to 30 years, and with increasing of age, the volume increases. 5. Immunohistochemically (with the expression of SD 34), a high rate of the number and area of ​​blood vessels detected in nodes at a young age, and a large diameter of blood vessels in patients at the age of 41-49, confirms the activation of angiogenesis at a young age with the formation of predominantly small-caliber arteries (arterioles) and capillaries, which is the basis for the active growth of nodes at a young age, in contrast to patients at the age of 41-49 with a high index of the average diameter of blood vessels; 6. The presence of small-diameter blood vessels indicates the maximum ability of the thinnest capillaries to reproduce and the possibilities to form anastomoses. Capillaries, as the thinnest tubes, represented by only one layer of endothelium, contribute to the activation of oxygen and metabolism in tissues and represent an important link in the molecular mechanisms of leiomyoma growth and development. 7. In line with the increase in the size of proliferative leiomyomas, the increase in the volume of the extracellular matrix with the decrease of angiogenesis and the development of hypoxia leads to create an obstacle for growth and development in the node with the onset of a remission period. 8. The high potential of self-renewal, differentiation, regeneration of blood vessels and constant active changes in the body of the uterus and within the node should be taken into consideration, as a hormone-dependent process in terms of the formation of new remodeled blood vessels, which is an important factor in the development of relapse. In addition, the active involvement of intranodal hypoxia and extracellular matrix in the process of remodeling blood vessels cannot be excluded.

Nodular leiomyoma of the uterus of giant size on the background of combined extragenital pathology (Case study)

The article presents a clinical case of nodular uterine leiomyoma of giant size on the background of type 2 diabetes mellitus, chronic hypertension and obesity with a classic picture of metabolic syndrome and the development of severe vascular pathology with steno-occlusive atherosclerotic lesions of the abdominal aorta bifurcation (Lerich’s syndrome) and acute circulatory disorders in the left lower extremity with gangrene of the toes of the left foot. The patient was hospitalized and comprehensively examined in the Department of Vascular Surgery of the Kyiv Regional Clinical Hospital (KRCH), which is the clinical base of the Department of Obstetrics and Gynecology №1 of the Shupyk National Healthcare University of Ukraine. The diagnosis was confirmed in the department and simultaneous two-stage surgery was successfully performed by gynecologists and vascular surgeons: first stage – pangysterectomy with revision of the abdominal organs and access to the abdominal aorta, second stage – thrombendarterectomy of the bifurcation of the abdominal aorta. In the postoperative period we restored peripheral blood flow, performed complete regression of ischemia, necrosis of the toes of the left foot dry, limited with a tendency to delimitation. Amputation of the first and second toes of the left foot was performed. Postoperative period was uneventful, on day 9 the patient was discharged home in satisfactory condition for further outpatient treatment. Diabetes mellitus type 2, chronic hypertension and obesity with a classic clinical picture of metabolic syndrome have led to the development of uterine leiomyoma and atherosclerotic changes in the abdominal aorta and main vessels of the lower extremities (Lerich’s syndrome). The successful outcome of this clinical case was made possible by the coordinated work of a multidisciplinary team of KRCH specialists both at the stage of comprehensive examination and diagnosis, and during simultaneous surgery.

Retrospective analysis of patients’ operative material during reproductive period with leiomyoma

Leiomyoma is known to be a benign tumor of unknown etiology of the uterus. It is most common in women of reproductive age with a cumulative frequency of 5.4% to 77.0%, and 80.0% of them suffer from this problem throughout their lives. However, some authors question the binding role of estrogen-progesterone in the growth of leiomyoma. However, due to the ineffectiveness of conservative treatment of leiomyoma, surgical intervention is still an effective method of curing the patient. There are no means of complete regression and prevention of this pathology. The aim of our study was retrospective analysis of operative material of patients of reproductive period with leukemia of the uterine body. Patients in both groups were divided into 40-year-olds and 41-50-year-olds according to age. The obtained results were processed using SPSS-21 software package of mathematical statistics. Pearson x2 criterion was used to test the reliability.Comparison of retrospective analysis of archival material revealed: A sharp increase in the incidence of intramural leiomyoma of the uterine body, due to many factors: social, ecological, stressful conditions, the introduction of modern diagnostic methods in practice. Proliferative, recurrent and latent leiomyomas are characterized by histostructural features. The process of growth of leiomyoma nodes is determined by the active products of the extracellular matrix, which leads to an increase in their size and does not depend on the histostructure, localization, number and size of the nodes Extracellular matrix products are important not only in terms of leiomyoma growth, but it is also likely to be among the factors that limit the free uncontrolled spread of tumor proliferation in the uterine body muscle and inhibit the assimilation process. This is confirmed by the absence of malignancy. Retrospective analysis of the material provides the basis for the search for molecular mechanisms of leiomyoma development. The results obtained will be important for the selection of drug treatment in terms of inhibition of myocyte proliferative processes, expression of growth factors and activation of apoptosis.