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Deletion of part of the long arm of chromosome 7 as revealed by a copy number variation array of a uterine leiomyoma. The red bar above the ideogram of chromosome 7 shows the deletion of 7q resulting from the hybridization. The blue dots in the upper part of the figure give an impression on the resolution of the methods. Of note, the deletion involves the CUX1 locus (gray vertical line). The numbers above the ideogram refer to the NCBI Build 37 reference sequence (hg19).
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Introduction:
Though uterine leiomyomas (UL) (syn.: fibroids) are by far the most frequent human symptomatic tumors their pathogenesis still remains to be elucidated. From the detection of microscopically visible alterations of chromosomal structure and molecular cytogenetic analyses, as well as from transcriptome and genome analyses, a picture of...
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Uterine fibroid presenting with acute abdominal pain is a rare event and thus imposes a diagnostic dilemma.
Leiomyomas are benign smooth muscle neoplasms originating from myometrium with an incidence as high as 70 to 80 percent in studies using histologic or sonographic examination. Acute abdominal pain is not a common symptom of fibroid and torsio...
Citations
... Among these, mutations in the gene encoding the RNA polymerase II (Pol II) transcriptional Mediator complex subunit 12 (MED12) are by far the most prevalent, accounting for approximately 70% of UFs (11). A proportionally smaller fraction of tumors are thought to arise from genetic alterations leading to overexpression of high mobility group AThook 2 (HMGA2) (approximately 20%), disruption of the collagen type 4 alpha 5/6 chain (COL4A5-COL4A6) locus (approximately 3%), or biallelic loss of fumarate hydratase (approximately 2%) (12)(13)(14). ...
Objective
To identify in myometrial stem/progenitor cells, the presumptive cell of origin for uterine fibroids, substrates of Mediator-associated CDK8/19 kinase, known to be disrupted by uterine fibroid driver mutations in Mediator subunit MED12.
Design
Experimental Study.
Setting
Academic Research Laboratory.
Patients
Women undergoing hysterectomy for uterine fibroids.
Interventions
Stable isotopic labeling of amino acids in myometrial stem/progenitor cell culture (SILAC) coupled with chemical inhibition of CDK8/19 and downstream quantitative phosphoproteomics and transcriptomic analyses.
Main Outcome Measures
High-confidence Mediator kinase substrates identified by SILAC-based quantitative phosphoproteomics were determined using an Empirical Bayes analysis and validated orthogonally by in vitro kinase assay featuring reconstituted Mediator kinase modules comprising wild-type or G44D mutant MED12 corresponding to the most frequent uterine fibroid driver mutation in MED12. Mediator kinase regulated transcripts identified by RNA-seq were linked to Mediator kinase substrates by computational analyses.
Results
296 unique phosphosites in 166 proteins were significantly decreased (> 2-fold; p < 0.05) upon CDK8/19 inhibition, including 118 phosphosites in 71 nuclear proteins representing high-confidence Mediator kinase substrates linked to RNA Polymerase II transcription, RNA processing and transport, chromatin modification, cytoskeletal architecture, and DNA replication and repair. Orthogonal validation confirmed a subset of these proteins, including CUX1 and FOXK1, to be direct targets of MED12-dependent CDK8 phosphorylation in a manner abrogated by the most common uterine fibroid driver mutation (G44D) in MED12, implicating these substrates in disease pathogenesis. Transcriptome-wide profiling of Mediator kinase-inhibited myometrial stem/progenitor cells revealed alterations in cell cycle and myogenic gene expression programs to which Mediator kinase substrates could be linked directly. Among these, CUX1 is an established transcriptional regulator of the cell cycle whose corresponding gene on chromosome 7q is the locus for a recurrent breakpoint in uterine fibroids, linking MED12 and Mediator kinase with CUX1 for the first time in uterine fibroid pathogenesis. FOXK1, a transcriptional regulator of myogenic stem cell fate, was found to be coordinately enriched along with kinase, but not core, Mediator subunits in myometrial stem/progenitor cells compared to differentiated uterine smooth muscle cells.
Conclusion
Altogether, these studies identify a new catalog of patho/biologically relevant Mediator kinase substrates implicated in the pathogenesis of MED12-mutation positive uterine fibroids, and further uncover a biochemical basis to link Mediator kinase activity with CUX1 and FOXK1 in the regulation of myometrial stem/progenitor cell fate.
... RAS, WNT/β-CANTENIN, TP53/ retinoblastoma protein (Rb), transforming growth factor beta (TGF-β) and nuclear factor kappa-light-chain-enhancer (NF-κB) signaling are affected by mutations in the MED12 gene which in turn have an impact on the downstream cellular processes that coordinate the DNA damage and repair responses [78]. Apparently, in UF, another target gene observed is CUX1 (Cut Like Homeobox 1, involved in chromosomal deletion) which encodes for a DNA binding protein of the homeodomain family and is known to overaccelerate repair of DNA damage, which can be error-prone [79]. Thus, these factors may be involved in inducing mutational heterogeneity in UF. ...
There has been a significant annual increase in the number of cases of uterine leiomyomas or fibroids (UF) among women of all races and ages across the world. A fortune is usually spent by the healthcare sector for fibroid-related treatments and management. Molecular studies have established the higher mutational heterogeneity in UF as compared to normal myometrial cells. The contribution of DNA damage and defects in repair responses further increases the mutational burden on the cells. This in turn leads to genetic instability, associated with cancer risk and other adverse reproductive health outcomes. Such and many more growing bodies of literature have highlighted the genetic/molecular, biochemical and clinical aspects of UF; none the less there appear to be a lacuna bridging the bench to bed gap in addressing and preventing this disease. Presented here is an exhaustive review of not only the molecular mechanisms underlying the predisposition to the disease but also possible strategies to effectively diagnose, prevent, manage, and treat this disease. Summary sentence Determining and designing of DNA damage response biomarker is needed urgently to predict uterine fibroid etiology, progression, and planning of non-invasive elimination measures to lessen the impact lead by this disease on the women related healthcare system and subsequently on the economy.
... [12][13] Several risk factors maybe responsible for UFs, including genetic changes, hormones, extracellular matrix, and other growth factors, such as tumor necrosis factor a (TNF-a), that may affect UFs growth. [14][15][16][17][18][19] A variety of studies reported that TNF-a is associated with UFs. [20][21][22][23][24] However, there is no systematic review exploring the association between TNF-a and UFs. ...
... [1][2][3] Many factors are reported to have association with UFs, such as TNF-a. [14][15][16][17][18][19] Many previous studies reported the association between TNF-a and UFs. [20][21][22][23][24] However, no systematic review has investigated this issue. ...
Background:
This study will explore the association between tumor necrosis factor α (TNF-α) and uterine fibroids (UFs).
Methods:
We will retrieve electronic databases in Cochrane Library, PUBMED, EMBASE, Web of Science, WANGFANG, Chinese Biomedical Literature Database, and China National Knowledge Infrastructure from inception to the present. All potential case-controlled studies investigating the association between TNF-α and UFs will be included in this study. Two researchers will independently select literature, appraise study quality, and extract outcome data. We will utilize a fixed-effects model or a random-effects model to synthesize outcome data. All data analysis will be performed by RevMan 5.3 software.
Results:
The present study will supply high-quality synthesis and/or descriptive analysis of the recent evidence to explore the association between TNF-α and UFs.
Conclusion:
This study will exert evidence to determine whether or not TNF-α is associated with UFs.
Study registration number:
INPLASY202070010.
... Although fibroids are monoclonal in origin (Linder and Gartler, 1965;Holdsworth-Carson et al., 2014), with tumours arising from a variety of genomic alterations in the stem cell of origin (MED12 and HMGA2 mutations are common) (Helmke et al., 2011;Mäkinen et al., 2011;Markowski et al., 2015), the histological presentation of a fibrotic phenotype is universal. Thus, targeting the genetic alterations is unlikely to yield treatments with general applicability without a labour intensive and expensive personalised medicine approach. ...
Current treatment options for uterine fibroids are limited to hormonal manipulation or surgical intervention. We aimed to develop an in-vitro model to mirror collagen deposition and extracellular matrix (ECM) formation, the principal features of uterine fibroids, to enable testing of novel therapeutics. Macromolecular crowding with Ficoll 400 and Ficoll 70 in cultures of human uterine myometrial smooth muscle cells containing ascorbic acid, provided the basis for this model. These culture conditions mimic the 'crowded' nature of the in-vivo extracellular environment by incorporating neutral, space-filling macromolecules into conventional cell cultures. This method of culture facilitates appropriate ECM deposition, thus closely representing the in-vivo fibrotic phenotype of uterine fibroids. Macromolecular crowding in Ficol cultures containing ascorbic acid reduced myometrial smooth muscle cell proliferation and promoted collagen production. Under these conditions, collagen was processed for extracellular deposition as demonstrated by C-propeptide cleavage from secreted procollagen. The fibrosis marker activin, was increased relative to its natural inhibitor, follistatin, in crowded culture conditions while addition of exogenous follistatin reduced collagen (Col1A1) gene expression. This in-vitro model represents a promising development for the testing of therapeutic interventions for uterine fibroids. However, it does not recapitulate the full in-vivo pathology which can include specific genetic and epigenetic alterations that have not been identified in the myometrial smooth muscle (hTERT-HM) cell line. Following screening of potential therapeutics using the model, the most promising compounds will require further assessment in the context of individual subjects including those with genetic changes implicated in fibroid pathogenesis.
... Current and well-established in vitro models in 2D and 3D growth format provide understanding of Minnie Malik and Joy Britten contributed equally to this work. [16][17][18][19][20]. They do not, however, provide insight into tumor-host interactions such as angiogenesis, physiologic impact of long-term growth, and other critical research areas only accessible via reliable in vivo models. ...
... Fibronectin (FN1) is a high-molecular weight ECM glycoprotein that binds to the cellular receptors, integrins, as well as ECM proteins and plays a major role in cell adhesion and growth [69,70]. Matrix protein fibronectin is highly expressed in leiomyoma tissue, and we have previously demonstrated its increased expression in leiomyoma tissue and 3D leiomyoma cultures as compared to myometrium [16,18,19]. Our 3D xenografts ( Figure 5-1K) demonstrated high levels of FN1 protein in the matrix similar to tissue xenografts ( Figure 5-1C). ...
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 mm³ 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.
... In summary, genetic studies on benign, borderline, and malignant smooth muscle tumors of uterine origin suggest a very low but existent probability of malignant transformation of initially benign tumors. A genetic classification of these lesions may be not only of diagnostic but also of predictive relevance (48)(49)(50)(51)(52) and should be mandatory for future clinical studies. ...
Our insights into the molecular pathogenesis of uterine smooth muscle tumors have improved significantly. Accordingly, in the present review, we advocate a more refined risk assessment for patients considering surgical removal of fibroids or hysterectomy, respectively, requiring morcellation. For this procedure, the risk estimates given for the iatrogenic spread of a previously unexpected malignancy considerably vary among different studies. Nearly all previous studies conducted retrospectively refer to the risk of a patient having an unexpected malignancy at the time of surgery. We feel that, more appropriately, risk should refer to the number of tumors because, as a rule, every single nodule arises independently and, thus, carries an independent risk of being malignant or not. Furthermore, whether so-called parasitic fibroids carry an underestimated risk of stepwise malignant transformation is discussed.
... For the course of growth of a myoma, more so factors on the molecular biological level (steroid receptors, growth factors) play a role [18,19], but perhaps also local changes of the myoma blood supply [13]. Furthermore, genetic factors seem to play an essential role [20]. This also underscores again the current recommendations to not draw upon the growth rate for posing a suspected diagnosis of uterus sarcoma. ...
Purpose:
Until now there are no systematic studies about the long-term course of myoma growth. Therefore, the aims of the present study were: (1) ultrasound monitoring of the natural course of growth of uterine leiomyomas; (2) assessment of whether the growth of myomas depends on the age of the patients, the location, or the initial size (possible co-factors/predictor criteria for increase of growth); influence of oral contraceptives (OC).
Methods:
Patient records (2010-May 2016) were retrospectively and systematically evaluated in regards to their growth and clinical course. The patients received a follow-up questionnaire by mail about the further history. Linear regression analysis and generalized regression analysis were performed to determine the influence of various factors on the growth of myomas.
Results:
Overall, 152 met the further inclusion criteria. Most of the myomas increased in size but 10% of the myomas became smaller without therapy. There is a significant dependency between the initial myoma size, and the first and second measurements, but not between those measures and myoma localization. In regression analysis, there was also a significant association between the growth of the myomas and the initial size but no association with age, complaint symptoms, and use of OC. However, the use of OC waas significantly associated with myoma growth in GEE.
Conclusions:
The course of growth of myomas has large variance, so this should not be taken as a sign for a malignant event (sarcoma or the so-called STUMP). The growth takes place with considerable individual variability and ultimately is not predictable.
Key Clinical Message
The patient was found to have multiple uterine myomas at the age of 19, underwent laparoscopic myomectomy at the age of 20, and underwent laparotomic myomectomy again at the age of 23 due to the recurrence of uterine myoma. At the age of 25, the patient reappeared with symptoms and recurrence, and was diagnosed with uterine leiomyomas (ULMs) of FH mutation and high‐grade squamous intraepithelial lesion (HSIL/CIN III) with gland involvement, after complete examination. Fumarate hydratase (FH) mutation screening is important when gynecologists encounter patients with early onset and multiple ULMs, it can give early diagnosis and treatment and fertility guidance. The patient had their uterus removed at the age of 26. FH mutation screening is important when gynecologists encounter patients with early onset and multiple ULMs, it can give early diagnosis and treatment and fertility guidance. It is also helpful for early diagnosis of renal cell carcinoma.
Subfertility, Reproductive Endocrinology and Assisted Reproduction - edited by Jane A. Stewart June 2019
