Uterine fibroids are the most common benign, monoclonal, gynecological tumors in a woman's uterus. To overcome the common obstacles related to the methods used in studying these pathologies, we aimed to devise a strategy to generate three-dimensional organoid models of myometrium and uterine fibroid stem cells using collagen-containing hydrogels as embedding scaffolds. Specifically, collagen-containing hydrogels in a low attachment V-bottom 96-well plates were exploited. This method allowed the development of 3D organoids of two stem cell types from normal myometrium and uterine fibroid-containing myometrium by embedding them in collagen-containing hydrogels and forming organoids in a suitable stem cell proliferation medium. The organoids successfully differentiated, proliferated, and self-organized into complex structures, developing a sustainable system. The importance of this model is the understanding of pathophysiology and etiopathogenesis, as well as for testing new drugs to prevent or treat uterine fibroids.

This study investigates how phthalate exposure contributes to uterine fibroid (UF) development by studying the effects of the Mono-(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), a metabolite of Di(2-ethylhexyl) phthalate, on myometrial stem cells (MMSCs). MMSCs from normal (MYON) and at-risk (MYOF) uterine tissues were cultured in 3D organoids and treated with 1.6 μM MEHHP for 48 h. Functional assays investigated cell viability, apoptosis, and mitochondrial activity, whereas RT-PCR, immunohistochemistry (IHC), and RNA sequencing evaluated markers of proliferation, apoptosis, extracellular matrix (ECM), and oxidative stress (OS). Cytokines and growth factors secretion were analyzed using a multiplex ELISA. Results showed that MEHHP exposure significantly increased cell viability and inhibited apoptosis in MYOF compared to MYON organoids. Proliferation markers (PCNA, Ki67), anti-apoptotic markers (BCL2/BAX ratio), and ECM markers (fibronectin and COL1A1) were significantly upregulated, whereas pro-apoptotic markers (Caspase-3) were downregulated in MYOF organoids. MEHHP-treated MYOF organoids exhibited elevated secretion of pro-inflammatory cytokines (e.g., TNF-α, IL-6, IL-8) and growth factors (e.g., PDGF, VEGF, TGFβ1), indicative of impaired tissue repair and fibrosis. RNA sequencing identified increased OS in MYOF organoids, validated by differential expression of genes such as CA9 and GPX3. Mitochondrial analysis revealed enhanced oxidative phosphorylation (OXPHOS) and elevated oxygen consumption rates, implicating mitochondrial dysfunction as a driver of cytokine release and UF pathogenesis. In conclusion, MEHHP was shown to promote the transformation of MYOF organoids into a UF phenotype by driving proliferation, inhibiting apoptosis, and inducing cytokine-mediated inflammation via mitochondrial dysfunction. These findings related to MYOF-specific effects, as compared to MYON, emphasize that these differences are statistically significant and relevant to UF risk. It can shed insight on how phthalates exposures may impact UF pathogenesis and provide a basis for exploring targeted therapeutic strategies.

INTRODUCTION: Uterine fibroid (UF) stiffness may contribute to tumor growth, symptom severity, and treatment response. Shear wave elastography (SWE) is a noninvasive, quantitative ultrasound method for measuring tissue stiffness, but it lacks standardization in gynecologic applications. This pilot study aimed to 1) develop a standardized transvaginal SWE protocol to assess myometrial and UF stiffness; 2) evaluate interrater and test–retest reliability; and 3) determine whether stiffness varies by menstrual phase. METHODS: In this prospective pilot study (January to September 2024), 27 women (17 with UFs, 10 without) were enrolled. Each underwent transvaginal SWE imaging during both the follicular and luteal phases of their menstrual cycle. Shear wave elastography values were independently calculated by two raters, with one rater repeating measurements after a 3-month interval to assess reliability. RESULTS: Fifty-one myometrial and 38 UF measurements demonstrated excellent interrater reliability (r=0.97, P <.001). Test–retest reliability was similarly high for 50 myometrial and 42 UF measurements (r=0.95 and r=0.98, respectively; P <.001). Uterine fibroids were significantly stiffer than the surrounding myometrium (median 52.4 versus 32.9 kPa, P <.001). Myometrial stiffness did not differ significantly between women with and without UFs ( P =.6), and menstrual phase had no effect on stiffness values. CONCLUSIONS/IMPLICATIONS: This pilot study establishes a reliable, reproducible protocol for transvaginal SWE in measuring uterine myometrial and UF tissue stiffness. These findings support its feasibility and potential value in personalized assessment and monitoring of UFs in clinical practice.

Uterine fibroids (UFs) are the most important benign neoplastic threat to women’s health worldwide, with no long-term noninvasive treatment options currently available. Among known UF driver alterations, somatic mutations in Mediator subunit MED12 are by the far the most prevalent, accounting for up to 80% of these clinically significant lesions. Although it is presently unclear how MED12 mutations trigger neoplastic transformation, MED12-mutant UFs are nonetheless characterized by significant chromosomal loss and rearrangement, suggesting genomic instability as a driving force in tumor development. However, the basis by which MED12 mutations drive genomic instability is not known. Herein, we show that R-loop-driven replication stress in MED12-mutant UFs leads to DNA under-replication and mitotic segregation errors that drive chromosomal instability. Notably, we find that vitamin D3 (VD3), a modifiable risk factor in UF development, suppresses pathogenic R-loop accrual and ameliorates replication stress-driven chromosomal instability, contributing to growth inhibition of patient-derived MED12-mutant UF xenografts in vivo. Altogether these findings uncover a molecular basis by which the predominant UF driver converges with a known risk factor at the interface of genomic instability, with significant translational implications for personalized UF prevention and treatment.

Chemical modifications of RNA add a dynamic regulatory layer to gene expression beyond the genome and epigenome. Among these modifications, 5-methylcytidine (m5C) has emerged as a key epitranscriptomic modification that influences RNA stability, translation, localization, and stress responses across diverse biological systems. Recent advances in high-resolution mapping and functional interrogation of m5C have revealed its involvement in development, metabolism, immune regulation, and disease pathogenesis. Notably, many of these processes are highly relevant to women’s health, which is shaped by hormone-responsive tissues, reproductive transitions, and pregnancy-associated physiological adaptations. In this review, we provide a comprehensive and integrative overview of m5C RNA modification with a focus on its roles in female biology and disease. We summarize the molecular machinery responsible for m5C deposition, recognition, and regulation, as well as current detection technologies. We further highlight emerging evidence linking m5C dysregulation to early embryonic development, women-specific cancers, gynecologic and reproductive disorders, pregnancy complications, and metabolic and cardiovascular diseases. In addition, we discuss the interplay between m5C and sex hormone signaling, as well as the potential of m5C as a biomarker and therapeutic target. Finally, we identify key knowledge gaps, including the need for tissue-specific, longitudinal, single-cell, and spatial epitranscriptomic studies in women. By integrating epitranscriptomics into women’s health research, this review underscores m5C as a previously underappreciated regulatory layer with significant implications for precision medicine and clinical translation.

Uterine fibroids are the most common benign gynecological tumors, affecting up to 70–80

Fertility preservation remains a significant concern for individuals undergoing gonadotoxic treatments. While traditional fertility preservation techniques are well-established, these methods can be time-consuming and limited by various medical or logistical barriers. In recent years, the potential of mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) has emerged as a promising, paradigm-shifting approach in fertility preservation. Preclinical studies have demonstrated the protective and regenerative properties of EVs in chemotherapy-induced ovarian and testicular damage in animal models. EVs provide a cell-free therapy that can potentially preserve ovarian function in females and spermatogenesis in males without the need for surgery or delay in cancer treatment. Additionally, using MSC-derived EVs offers advantages over traditional stem cell therapies, such as a reduced risk of immune rejection, targeted treatment, and avoidance of safety concerns associated with stem cell-based therapies. Future directions include enhancing the therapeutic potential of MSC-derived EVs through genetic engineering or cell priming techniques to target specific tissues and further optimize their utilization in fertility preservation. Given the potential of MSC-derived EVs to protect fertility in both females and males, this approach could revolutionize treatment in oncofertility. Further research, including clinical trials, is necessary to confirm the safety and efficacy of MSC-derived EVs, focusing on premature ovarian insufficiency. Looking ahead, MSC-derived EVs could revolutionize fertility preservation, offering hope for cancer patients and individuals exposed to various environmental risks affecting reproduction, including in space exploration, where protection from cosmic radiation is essential.

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Purpose:Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder in reproductive-age women. Several published studies demonstrated the therapeutic potential of vitamin D on PCOS condition, but vitamin D supplementation leads to only modest improvements. Doxercalciferol (Dox) is a highly potent vitamin D receptor (VDR) agonist that binds to VDR with at least 150 times greater affinity than other vitamin D compounds. In this study, we hypothesized that the therapeutic potential of Dox may reverse PCOS-related phenotypes more efficiently. Method:We first analyzed the regulatory potential of Dox in fat metabolism by comparing adipocyte proliferation and differentiation in our in vitro model. We also injected Dox into our mouse model of PCOS to analyze the therapeutic effect of Dox treatment on various PCOS-related parameters and fertility. Result:Dox treatment inhibited the proliferation of both adipose lineage cells and androgen-producing H295R cells in a dose-dependent manner. PCOS-related outcomes such as body weight and fertility rate were significantly reversed in Dox-treated PCOS mice model. Conclusion:Our study revealed that vitamin D analogs are a promising family of compounds for reducing body weight and reversing infertility in an animal model of PCOS.

Uterine leiomyomas are characterized by excessive extracellular matrix (ECM) deposition, enhanced proliferative capacity, and progressive tissue remodeling. Emerging evidence suggests that advanced glycation end-products (AGEs), key mediators of oxidative and metabolic stress, contribute to fibroid pathogenesis; however, the mechanistic role of the AGE/receptor for advanced glycation end-products (RAGE) axis remains poorly understood. Here, we investigated the pathogenic significance of RAGE signaling and evaluated the therapeutic potential of the selective RAGE antagonist FPS-ZM1 in uterine leiomyomas. Clinical transcriptomic datasets (GSE128229 and GSE13319) and human leiomyoma specimens demonstrated elevated RAGE expression relative to matched myometrium tissues. In ELT-3 leiomyoma cells, AGE stimulation promoted proliferative capacity, clonogenicity, tumorsphere formation, and extracellular matrix accumulation, accompanied by increased expression of fibrosis- and stemness-associated markers. Quantitative proteomic analyses revealed that AGE exposure altered pathways associated with fatty acid β-oxidation, mitochondrial function, focal adhesion, and cytoskeletal organization, which were partially normalized following FPS-ZM1 treatment. Mechanistically, AGE stimulation altered mitochondrial homeostasis and mitochondrial dynamics, activated MAPK signaling, and modulated autophagy-associated pathways. Pharmacological inhibition of RAGE attenuated these pathological alterations and reduced oxidative stress-associated responses. In vivo, FPS-ZM1 significantly suppressed leiomyoma xenograft growth and reduced RAGE/HMGB1 expression in tumor tissues. Collectively, our findings identify the AGE/RAGE axis as an important regulator of metabolic remodeling, mitochondrial alterations, and fibrotic progression in uterine leiomyomas. Targeting RAGE signaling may represent a promising non-hormonal therapeutic strategy for uterine fibroid management.

Uterine fibroids (UFs) are highly prevalent benign tumors, primarily affecting women of reproductive age. The prevalence of UFs is estimated at 25% to 70% to 80%, with significantly higher rates among African women. Recent investigations have unveiled a link between vitamin D (VD) levels and the presence of UFs. Current data show that individuals with sufficient VD levels are at a reduced risk of developing UFs. This review aims to synthesize the latest discoveries on the involvement of VD in the pathophysiology of UFs, as well as to explore its feasible therapeutic applications. The mechanisms underlying the potential of VD to diminish the risk of UF development and to inhibit their growth are multifaceted and include its antiproliferative effects, the induction of apoptosis, suppression of angiogenesis, or the modulation of enzyme activity. The potential of VD and its analogues as promising agents for managing and preventing UFs was demonstrated. However, further research is needed to clarify the biological mechanisms behind the role of VD in UF pathophysiology, and its definitive efficacy has to be confirmed through randomized clinical trials.

Endometriosis is a chronic, systemic, oestrogen-dependent disease with major impacts on pain, fertility and quality of life. This Society of Endometriosis and Uterine Disorders and American Institute for Minimally Invasive Surgery opinion synthesizes current evidence to redefine diagnostic and therapeutic strategies across the continuum of care. Diagnostic laparoscopy is no longer indicated, as clinical assessment combined with expert transvaginal ultrasonography and magnetic resonance imaging reliably identifies disease phenotypes and guides management. Medical therapy, particularly progestins, remains the foundation of long-term care, with gonadotrophin-releasing hormone analogues and antagonists as second-line options. Surgery is reserved for clearly defined indications - including deep infiltrating disease, infertility with correctable anatomy and organ-threatening involvement - and must be preceded by comprehensive imaging and followed by prolonged hormonal suppression to reduce recurrence. Integrated, individualized care pathways are essential to improve outcomes and support fertility preservation. This framework represents a modern paradigm for comprehensive endometriosis management.