It is well established that the vagina is colonized by bacteria that serve important roles in homeostasis. Imbalances in the proportion of bacteria may lead to a predisposition to infection or ...reproductive complications. Molecular-based approaches demonstrated a greater degree of microbial diversity both within and between women than previously recognized. The vaginal microbiome may fluctuate during various states of health, such as during the menstrual cycle or after menopause, and there may be differences in the vaginal microbiome between women of different ethnicities. Furthermore, the specific composition of the vaginal microbiome may influence the predisposition to dysbiosis and the transmission of sexually transmitted infections. An understanding of the diversity of the vaginal microbial environment during states of health is essential for the identification of risk factors for disease and the development of appropriate treatment.
Fibroids are benign uterine tumors characterized by the proliferation of uterine smooth muscle cells, embedded in an abundant extracellular matrix. Their prevalence is estimated to be >50% in women ...aged >45 years. Fibroids represent a considerable health burden. It is time to acquire a deeper mechanistic understanding of uterine fibroid-related etiology and pathogenesis, which may help pinpoint new strategies and an individualized approach. There is a need to gather prospective data and conduct studies to compare alternative approaches and assess long-term outcomes with respect to quality of life, recurrence of symptoms (bleeding and bulk symptoms), fertility, and even complications The goal of this review was to evaluate the widely accepted pathogenesis and identify risks factors and future directions for clinical and basic research into fibroids.
Uterine leiomyomas are the most common benign gynecologic condition. The prevalence is three times more common among women of African ethnicity. Disparity in this disease is evidenced by earlier age ...of onset, greater severity of symptoms, and different response to treatment. Although the pathogenesis of disease development is not completely known, growing evidence focuses on investigating the molecular mechanisms in disease development and the influence of ethnicity. Variation in the expression levels or function of estrogen and progesterone receptors, polymorphism of genes involved in estrogen synthesis and/or metabolism (COMT, CYP17), retinoic acid nuclear receptors (retinoid acid receptor-α, retinoid X receptor-α), and aberrant expression of micro-RNAs (miRNAs) are some of the molecular mechanisms that may be involved. Nutritional factors, such as vitamin D deficiency, might also contribute to the higher incidence in dark skinned populations who are also commonly suffer from hypovitaminosis D. Culture and environmental difference might have a role in disease development. Further analysis and better understanding of these mechanisms will provide insight into the molecular basis of racial disparities in leiomyoma formation and will help to develop new innovations in leiomyoma treatment.
Abstract Background Uterine leiomyoma is the most common pelvic tumor in women, but the actual prevalence is unknown. Objectives To review the literature on the prevalence of uterine leiomyoma, ...presenting symptoms, and medical management. Search strategy On April 1–30, 2014, a PubMed search for studies reported in English was conducted using the terms “uterine leiomyoma,” “prevalence,” and “symptoms.” Another search was performed using the terms “uterine leiomyoma” and “treatment.” Selection criteria All trial types other than internet-only studies were included. Animal studies were excluded from the prevalence/symptom review, but included in the medical management review. Data collection and analysis Prevalence rates were recorded on the basis of imaging modality, cohort studied, ethnic origin, and age. Main results Studies involving asymptomatic women revealed a trend in prevalence similar to that in symptomatic women, and showed that leiomyomas are more common in this cohort than previously recognized. Affected patients can present with many complaints, but no single symptom has been shown to be specific for this tumor. Various medical therapies are reviewed, summarizing efficacy and toxicity. Conclusions Further research needs to be conducted on the prevalence in asymptomatic women. Current and future medical management options provide promising results in symptom reduction.
To observe the antifibroid effects of therapeutic concentrations of simvastatin, which interferes with cholesterol biosynthesis, a known precursor of five major classes of steroid hormones, including ...progesterone and estrogen, which play a major role in the development and growth of uterine leiomyomas.
Two-dimensional and three-dimensional cell culture study of immortalized human leiomyoma and patient-matched myometrium cells treated with simvastatin.
University laboratory.
None.
None.
Cell proliferation, alteration in apoptotic signaling pathways, and extracellular matrix (ECM) protein production.
Simvastatin demonstrated a concentration-dependent antiproliferative effect on both the leiomyoma cells and the patient-matched myometrium cells, but a higher inhibitory effect at lower concentrations of simvastatin was observed in leiomyoma cells. Simvastatin also regulated leiomyoma cell apoptosis through a concentration-dependent increase in activity of caspase-3. Simvastatin significantly inhibited expression of major ECM proteins collagen I, collagen III, fibronectin, versican, and brevican in leiomyoma cells at concentrations as low as 10−9 mol/L within 48 hours of exposure.
Simvastatin induces apoptosis in uterine leiomyoma cells at low concentrations, as evidenced by increased active caspase levels. Furthermore, inhibited production of the ECM proteins may lead to reduction in tumor size. Simvastatin may represent a novel therapeutic treatment strategy for uterine leiomyomas.
La simvastatina a concentraciones clínicamente relevantesafecta el crecimiento del leiomioma uterino humano y la producción de matriz extracelular
observar los efectos antifibroides de las concentraciones terapéuticas de simvastatina, las cuales interfieren con la biosíntesis del colesterol, un conocido precursor de las cinco clases principales de hormonas esteroides, incluida la progesterona y el estrógeno, los cuales juegan un papel importante en el desarrollo y crecimiento de los leiomiomas uterinos.
estudio de cultivo celular bidimensional y tridimensional de leiomioma humano inmortalizado y células miometriales de pacientes tratados con simvastatina.
Laboratorio Universitario.
Ninguno.
Ninguna.
Proliferación celular, alteración en las vías de señalización apoptótica y producción de proteína de matriz extracelular (MEC).
La simvastatina demostró un efecto antiproliferativo dependiente de la concentración tanto en las células de leiomioma como en las células miometriales emparejadas por paciente, pero se observó un mayor efecto inhibidor sobre las células de leiomioma con bajas concentraciones de simvastatina. La simvastatina también reguló la apoptosis de las células leiomiomas a través de un aumento dependiente de la concentración en la actividad de la caspasa-3. La simvastatina inhibió significativamente la expresión de las principales proteínas de la MEC colágeno I, colágeno III, fibronectina, versican y brevican en células de leiomioma en concentraciones tan bajas como 10 -9 mol / L dentro de las 48 horas posteriores a la exposición.
La simvastatina a bajas concentraciones induce apoptosis en células de leiomioma uterino, como lo demuestra el aumento de los niveles de caspasa activa. Además, la producción inhibida de las proteínas MEC puede provocar una reducción en el tamaño del tumor. La simvastatina puede representar una nueva estrategia de tratamiento terapéutico para los leiomiomas uterinos.
Uterine leiomyomas, the most common tumors of the female reproductive system, are characterized by excessive deposition of disordered stiff extracellular matrix and fundamental alteration in the ...mechanical signaling pathways. Specifically, these alterations affect the normal dynamic state of responsiveness to mechanical cues in the extracellular environment. These mechanical cues are converted through integrins, cell membrane receptors, to biochemical signals including cytoskeletal signaling pathways to maintain mechanical homeostasis. Leiomyoma cells overexpress β1 integrin and other downstream mechanical signaling proteins. We previously reported that simvastatin, an antihyperlipidemic drug, has antileiomyoma effects through cellular, animal model, and epidemiologic studies.
This study aimed to examine the hypothesis that simvastatin might influence altered mechanotransduction in leiomyoma cells.
This is a laboratory-based experimental study. Primary leiomyoma cells were isolated from 5 patients who underwent hysterectomy at the Department of Gynecology and Obstetrics of the Johns Hopkins University Hospital. Primary and immortalized human uterine leiomyoma cells were treated with simvastatin at increasing concentrations (0.001, 0.01, 0.1, and 1 μM, or control) for 48 hours. Protein and mRNA levels of β1 integrin and extracellular matrix components involved in mechanical signaling were quantified by quantitative real-time polymerase chain reaction, western blotting, and immunofluorescence. In addition, we examined the effect of simvastatin on the activity of Ras homolog family member A using pull-down assay and gel contraction.
We found that simvastatin significantly reduced the protein expression of β1 integrin by 44% and type I collagen by 60% compared with untreated leiomyoma cells. Simvastatin-treated cells reduced phosphorylation of focal adhesion kinase down to 26%–60% of control, whereas it increased total focal adhesion kinase protein expression. Using a Ras homolog family member A pull-down activation assay, we observed reduced levels of active Ras homolog family member A in simvastatin-treated cells by 45%–85% compared with control. Consistent with impaired Ras homolog family member A activation, simvastatin treatment reduced tumor gel contraction where gel area was 122%–153% larger than control. Furthermore, simvastatin treatment led to reduced levels of mechanical signaling proteins involved in β1 integrin downstream signaling, such as A-kinase anchor protein 13, Rho-associated protein kinase 1, myosin light-chain kinase, and cyclin D1.
The results of this study suggest a possible therapeutic role of simvastatin in restoring the altered state of mechanotransduction signaling in leiomyoma. Collectively, these findings are aligned with previous epidemiologic studies and other reports and support the need for clinical trials.
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Multiple in vivo animal models for uterine leiomyoma do not adequately represent human disease based on etiology, molecular phenotype, or limited fixed life span. Our objective was to develop a ...xenograft model with sustained growth, by transplanting a well-established actively growing three-dimensional (3D) cell culture of human leiomyoma and myometrium in NOD/SCID ovariectomized female mice. We demonstrated continued growth to at least 12 weeks and the overexpression of extracellular matrix (ECM). Further, we confirmed maintenance of hormonal response that is comparable to human disease in situ. Leiomyoma xenografts under hormonal treatment demonstrated 8 to12-fold increase of volume over the xenografts not treated with hormones. Estradiol-treated xenografts were more cellular as compared to progesterone or combination milieu, at the end of 8-week time frame. There was also a non-statistically significant 2-4 mm
increase in volume between 8-week and 12-week xenografts with higher matrix to cell ratio in 12-week xenografts compared to the 8-week and placebo xenografts. Increased expression of ECM proteins, fibronectin, versican, and collagens, indicated an actively growing cell matrix formation in the xenografts. In conclusion, we have developed and validated a xenograft in vivo model for uterine leiomyoma that shares the genomic and proteomic characteristics with the human surgical specimens of origin and recapitulates the most important features of the human tumors, the aberrant ECM expression that defines the leiomyoma phenotype and gonadal hormone regulation. Using this model, we demonstrated that combination of estradiol and progesterone resulted in increased cellularity and ECM production leading to growth of the xenograft tumors.
To characterize the effect of ulipristal acetate (UPA) treatment on transforming growth factor (TGF) canonical and noncanonical signaling pathways in uterine leiomyoma tissue and cells. UPA decreased ...extracellular matrix in surgical specimens; we characterize the mechanism in this study.
Laboratory study.
University.
Exposure of leiomyoma cell lines to UPA.
RNAseq was performed on matched myometrium and leiomyoma surgical specimens of placebo- and UPA-treated patients. Changes in gene expression and protein were measured using quantitative polymerase chain reaction and western immunoblot analysis, respectively.
In surgical specimen, mRNA for TGF-β3 was elevated 3.75-fold and TGFR2 was decreased 0.50-fold in placebo leiomyomas compared with myometrium. Analysis of leiomyomas from UPA-treated women by western blot revealed significant reductions of active TGF-β3 (0.64 ± 0.12-fold), p-TGFR2 (0.56 ± 0.23-fold), pSmad 2 (0.54 ± 0.04-fold), and pSmad 3 (0.65 ± 0.09-fold) compared with untreated leiomyomas. UPA treatment demonstrated statistically significant reduction in collagen 1, fibronectin, and versican proteins. Notably, there was a statistically significant increase of the extracellular matrix protein fibrillin in leiomyoma treated with UPA (1.48 ± 0.41-fold). Data from in vitro assays with physiologic concentrations of UPA supported the in vivo findings.
TGF-β pathway is highly up-regulated in leiomyoma and is directly responsible for development of the fibrotic phenotype. UPA attenuates this pathway by reducing TGF-β3 message and protein expression, resulting in a reduction in TGF-β canonical signaling. In addition, UPA significantly increased fibrillin protein expression, which can serve to bind inactive TGF-β complexes. Therefore, UPA inhibits leiomyoma fibrosis by decreasing active TGF-β3 and diminishing signaling through the canonical pathway.
NCT00290251.
El acetato de ulipristal disminuye el TGF-b3 activo y su vía de señalización canónica en leiomiomas uterinos a través de dos nuevosmecanismos
Caracterizar el efecto del tratamiento con acetato de ulipristal (UPA) sobre el factor de crecimiento transformante (TGF) a través de sus vías de señalización canónicas y no canónicas tanto en tejido como en células procedentes de leiomiomas uterinos. UPA disminuyó la matriz extracelular en muestras quirúrgicas; en este estudio caracterizamos el mecanismo.
Estudio de laboratorio.
Universidad.
Exposición de líneas celulares de leiomioma a UPA.
Se realizó RNAseq en muestras quirúrgicas pareadas de miometrio y leiomiomas procedentes de pacientes con efecto placebo y tratadas con UPA. Los cambios en la expresión génica y proteica se midieron utilizando la reacción en cadena de la polimerasa cuantitativa y el análisis de inmunotransferencia Western blot, respectivamente.
En las muestras quirúrgicas, el ARNm para TGF-b3 se elevó 3,75 veces y para TGFR2 disminuyó 0,50 veces en los leiomiomas con efecto placebo en comparación con el miometrio. El análisis por Western blot de los leiomiomas de mujeres tratadas con UPA indicó reducciones significativas de TGF-b3 activo (0,64 ± 0,12), p-TGFR2 (0,56 ± 0,23), pSmad 2 (0,54 ± 0,04) y pSmad 3 (0,65 ± 0,09) en comparación con los leiomiomas no tratados. El tratamiento con UPA demostró una reducción estadísticamente significativa a nivel proteico del Colágeno 1, la Fibronectina y el Versican. En particular, hubo un aumento estadísticamente significativo de la proteína de la matriz extracelular Fibrilina en leiomiomas tratados con UPA (1,48 ± 0,41). Los datos de los ensayos in vitro con concentraciones fisiológicas de UPA respaldaron los resultados obtenidos in vivo.
La vía del TGF-b está altamente regulada en leiomiomas y es directamente responsable del desarrollo del fenotipo fibrótico. UPA atenúa esta vía al reducir la expresión TGF-b3 (mensajero y proteínas), lo que resulta en una reducción en la vía de señalización canónica del TGF-b. Además, UPA aumentó significativamente la expresión de la proteína fibrilina, la cual puede servir para unir complejos de TGF-b inactivos. Por lo tanto, UPA inhibe la fibrosis de los leiomiomas al disminuir el TGF-b3 activo y disminuir la señalización a través de la vía canónica.
NCT00290251.
Uterine leiomyomas are benign tumors in the smooth muscle layer of the uterus. The most common histological type is the “usual leiomyoma”, characterized by overexpression of ECM proteins, whereas the ...“cellular type” has higher cellular content. Our objective is to investigate the involvement of inflammatory and reparative processes in leiomyoma pathobiology. Using a morphological approach, we investigate the presence of inflammatory cells. Next, we determine the localization of the ECM, the presence/absence of fibrotic cells via α-sma and desmin and the immunohistochemical profile of the mesenchymal cells with respect to CD34. Finally, we explore the effect of inflammatory mediators (TNF-α, IL-1β, IL-6, IL-15, GM-CSF and IFN-γ) on pro-fibrotic factor activin A mRNA expression in vitro. Higher numbers of macrophages were found inside and close to leiomyomas as compared to the more distant myometrium. Cellular leiomyomas showed more macrophages and mast cells than the “usual type”. Inside the fibroid tissue, we found cells positive for α-sma, but negative for desmin and a large amount of collagen surrounding the nodule, suggestive of myofibroblasts producing ECM. In the myometrium and leiomyomas of the “usual type”, we identified numerous CD34+ fibroblasts, which are known to give rise to myofibroblasts upon loss of CD34 expression. In leiomyomas of the “cellular type”, stromal fibroblasts were CD34-negative. Finally, we found that TNF-α increased activin A mRNA in myometrial and leiomyoma cells. In conclusion, this study demonstrates the presence of inflammatory cells in uterine leiomyomas, which may contribute to excessive ECM production, tissue remodeling and leiomyoma growth.
Uterine fibroids (UFs) are the most common benign gynecological tumors. It was estimated that fifty percent of women presenting with UFs has symptomatology that negatively influences their quality of ...life. Pharmacological and/or surgical treatments are frequently required, depending on the woman's desire to preserve fertility, with a high impact on healthcare costs. Generally, the use of currently available pharmacological treatments may lead to side effects. Therefore, there is a growing interest in a natural and safe approach for UFs. In recent years, epidemiological studies reported a vitamin D deficiency in patients with UFs raised interest in the potential biological effects of vitamin D supplementation. In vitro studies proved vitamin D efficacy in inhibiting UFs growth by targeting pathways involved in the regulation of various biological processes, including proliferation, extracellular matrix (ECM) remodeling, DNA repair, signaling and apoptosis. However, clinical studies supported only in part the beneficial effects of vitamin D supplementation in reducing UFs growth and tumor volume. Randomized controlled trials and large population studies are mandatory as the potential clinical benefits are likely to be substantial.
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