RARE DISEASE
Malignant granulosa cell tumor of the ovary
Malignant granulosa cell tumor of the ovary
Malignant granulosa cell tumor of the ovary
Synonyms: Granulosa cell cancer, Granulosa cell malignant tumor
Synonyms: Granulosa cell cancer, Granulosa cell malignant tumor
Synonyms: Granulosa cell cancer, Granulosa cell malignant tumor
Drug discovery
2
drugs
With orphan designations
Overview
Malignant granulosa cell tumor (GCT) of the ovary is a rare sex cord-stromal neoplasm arising from granulosa cells, primarily affecting peri- and postmenopausal women [1][2]. Characterized by FOXL2 mutations (95% of cases) [9], these estrogen-secreting tumors often present with abnormal vaginal bleeding, abdominal pain, or palpable masses [1][2]. While typically indolent with a favorable prognosis in early stages, they exhibit a propensity for late recurrence (up to 20 years) [1][4][9]. Diagnosis combines histopathology (Call-Exner bodies, nuclear grooves) with elevated serum inhibin [2][9]. Surgical resection is first-line therapy, with adjuvant chemotherapy for advanced disease [4][7][10].
Therapies
Surgery: Primary treatment with staging/debulking; fertility-sparing approaches for early-stage disease [4][7][10]
Chemotherapy: BEP regimen (bleomycin, etoposide, cisplatin) for advanced/metastatic disease (20–60% response rates) [3][7][10]
Targeted/hormonal therapy: Aromatase inhibitors, GnRH analogs, or anti-progestins in recurrent/refractory cases [10][13]
Categories: rare gynecological and obstetric diseases, rare neoplastic diseases
Research Papers
894 drug discovery papers about Malignant granulosa cell tumor of the ovary, with 2 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
894 drug discovery papers about Malignant granulosa cell tumor of the ovary, with 2 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
categories:
Small molecules
small molecules
2026-09-26 | Treatment Outcomes and Toxicity of Cisplatin-Etoposide-Based Regimens and Carboplatin-Paclitaxel as Adjuvant Chemotherapy for Ovarian Adult Granulosa Cell Tumors: A Multicenter Retrospective Cohort Study.
Background/Objectives: Adult granulosa cell tumors (AGCTs) of the ovary are rare sex cord-stromal malignancies characterized by an indolent clinical course and a persistent risk of late recurrence. Although adjuvant chemotherapy is considered for selected patients with higher-risk disease, the optimal platinum-based regimen remains uncertain because comparative evidence is limited. This study compared disease-free survival (DFS), treatment delivery, and toxicity between bleomycin-etoposide-cisplatin/etoposide-cisplatin (BEP/EP) and carboplatin-paclitaxel (CP) in the postoperative adjuvant setting. Methods: This retrospective, multicenter, nonrandomized comparative cohort study included 58 patients with AGCTs who underwent primary surgery and received adjuvant platinum-based chemotherapy at five tertiary oncology centers in Türkiye between 2010 and 2024. Patients received BEP/EP or CP according to institutional practice and physician discretion. The primary endpoint was disease-free survival (DFS), while secondary endpoints included treatment delivery and treatment-related toxicity. Results: A total of 58 patients were included, with 26 receiving BEP/EP and 32 receiving CP. Patients treated with CP were older, whereas fertility-sparing surgery was more common in the BEP/EP group. After a median follow-up of 75.5 months, 14 recurrences were observed; when deaths without prior recurrence were included according to the prespecified DFS definition, a total of 19 DFS events occurred. Median DFS was 77.2 months in the BEP/EP group and 126.2 months in the CP group. The estimated 5-year DFS rates were 75.5% and 83.5%, respectively, with no statistically significant difference between treatment groups (log-rank p = 0.296). In an exploratory multivariable Cox model restricted to age and treatment regimen, increasing age was associated with a higher risk of a DFS event (adjusted HR 1.046 per year, 95% CI 1.011-1.082; p = 0.010), whereas CP versus BEP/EP was not significantly associated with DFS (adjusted HR 0.426, 95% CI 0.167-1.083; p = 0.073). Treatment completion rates were numerically higher with CP (84.4% vs. 69.2%; p = 0.17). Peripheral neuropathy occurred more frequently with CP (p = 0.007), whereas nephrotoxicity occurred only in the BEP/EP group (p = 0.014). Pulmonary toxicity was observed only with bleomycin-containing treatment, and one patient developed secondary acute myeloid leukemia following BEP. Conclusions: No statistically significant difference in DFS was detected between BEP/EP and CP, while distinct toxicity patterns were observed. Given the retrospective design, limited sample size, and potential for treatment-selection bias and residual confounding, these findings should be considered exploratory and should not be interpreted as evidence of equivalent efficacy between the regimens.
2026-09-22 | Docetaxel for recurrent adult granulosa cell tumor enabling tertiary cytoreduction
Background Adult granulosa cell tumors (AGCTs) are rare ovarian sex cord-stromal tumors characterized by an indolent clinical course, frequent late recurrences, and limited evidence to guide systemic therapy for recurrent disease. Although platinum- and taxane-based regimens have demonstrated activity, optimal treatment after multiple recurrences remains undefined. Case We report a 53-year-old woman with recurrent AGCT who had progressed on multiple chemotherapy and hormonal regimens over eight years. Despite prior exposure to and progression on paclitaxel, her disease responded to single-agent docetaxel for 36 cycles, reducing disease burden sufficiently to enable tertiary cytoreductive surgery. The treatment was well tolerated. Conclusion This case highlights the potential activity of docetaxel in recurrent AGCT and supports its consideration as an alternative taxane, particularly for patients who are not candidates for or have progressed after other systemic therapies including paclitaxel. Reports describing the use of docetaxel in AGCT are limited, with only isolated cases published to date. Given the rarity of AGCT, individual case reports remain a valuable source of evidence to inform treatment decisions. Prospective studies are needed to better define the role of docetaxel in recurrent AGCT.
2026-08-27 | Isorhamnetin and Female Reproduction: Effects on Viability, Hormone Secretion, and Growth Factors.
Background/Objectives: Isorhamnetin is a naturally occurring flavonoid with reported antioxidant, anti-inflammatory, and anticancer properties. Although its biological activities have been extensively investigated, its effects on ovarian cell physiology remain insufficiently characterized. This study aimed to evaluate the influence of isorhamnetin on the viability, steroid hormone secretion, apoptosis, and growth factor signalling in human ovarian cell lines representing both non-tumour and tumour phenotypes. Methods: Human granulosa (HGL5), granulosa tumour (COV434), and epithelial ovarian carcinoma (OVCAR-3) cell lines were treated with isorhamnetin at concentrations of 5-80 μg/mL for 24 h. Cell viability was determined using the AlamarBlueTM assay. The secretion of progesterone, 17β-estradiol, and the presence of transforming growth factor β2 (TGF-β2), transforming growth factor β receptor 2 (TGFBR2), and apoptosis-inducing factor (AIF) was quantified by enzyme-linked immunosorbent assay (ELISA). Results: Isorhamnetin significantly reduced cell viability in a dose-dependent manner, with tumour cell lines exhibiting greater sensitivity than non-tumour granulosa cells. A significant decrease in viability was observed in OVCAR-3 cells from 10 μg/mL onward, whereas COV434 cells showed significant reductions at concentrations of 20 μg/mL and higher. In contrast, HGL5 cell viability was significantly affected only at the highest concentration (80 μg/mL). No significant changes were detected in the secretion of progesterone, 17β-estradiol, TGF-β2, or TGFBR2 in any of the examined cell lines. A significant reduction in AIF production was observed only in HGL5 cells treated with 80 μg/mL isorhamnetin. Conclusions: Isorhamnetin preferentially decreased the viability of ovarian tumour cells while exerting only limited effects on non-tumour granulosa cells, indicating preferential cytotoxic activity toward malignant ovarian cells. The absence of significant changes in steroid hormone production and TGF-β suggests that its antiproliferative effects involve molecular pathways that were not investigated in the present study. These findings support further investigation of isorhamnetin as a potential natural compound for ovarian cancer prevention or adjunctive therapy.
2026-08-11 | A phase II study of androgen receptor inhibition by darolutamide in combination with leuprolide acetate and exemestane in recurrent adult-type ovarian granulosa cell tumor.
To determine the efficacy and safety of darolutamide, exemestane, and leuprolide acetate in recurrent adult-type ovarian granulosa cell tumor (AGCT). This single-arm Phase II cooperative group study included patients with recurrent AGCT who had progressed on a prior aromatase inhibitor. The treatment regimen consisted of darolutamide 600 mg by mouth twice daily, exemestane 25 mg by mouth once daily, and leuprolide acetate 7.5 mg by intramuscular injection every four weeks. Patient accrual occurred from January 2024 to April 2024 with sample size determined using Simon's Optimal two-stage design. The primary measure of efficacy was objective response rate (ORR) measured by Response Evaluation Criteria in Solid Tumors 1.1. Secondary objectives included duration of response, progression-free survival (PFS), overall survival (OS), and safety of the treatment regimen. Seventeen patients were enrolled to complete target accrual of the first stage. Of the 16 evaluable patients, there was one partial response (ORR = 6.25%). The trial did not continue to the second stage as it did not meet its prespecified endpoint. Ten patients (62.5%) were confirmed to have stable disease and five patients (31.25%) were confirmed to have progressive disease. Median PFS was 8.5 months (95% CI: 3.1 months - 12.0 months). Median OS was not reached. There were no grade 4 or grade 5 adverse events attributable to the treatment regimen. Though the trial did not meet its primary endpoint, there was one partial response to the treatment regimen. Clinically meaningful PFS was demonstrated. Clinical Trial Registration Number NCT06169124.
2026-08-02 | Citrus flavonoids mitigate the cisplatin-induced ovarian toxicity via dual modulation of Nrf2/HO-1 pathway and NF-κB axis.
Cisplatin (CIS) is an effective chemotherapeutic agent whose clinical use is limited by off-target toxicity, including damage to ovarian granulosa cells, characterized by oxidative stress and inflammation. Here, the cytoprotective potential of the citrus flavonoids, naringin (NG) and hesperidin (HP), was investigated in ovarian granulosa cells exposed to CIS-induced toxicity. Molecular docking demonstrated that NG and HP exhibited strong binding affinities for key antioxidant, nuclear factor erythroid 2-related factor 2 (Nrf2) and catalase (CAT) as well as anti-apoptotic B-cell lymphoma-2 (Bcl-2) proteins, while CIS preferentially interacted with pro-apoptotic targets. In vitro, CIS dose-dependently reduced cell viability and hormone secretion (progesterone and estradiol), and these reductions were significantly restored by co-treatment with NG or HP. CIS-induced oxidative injury, marked by elevated accumulation of reactive oxygen species (ROS), lipid peroxidation, and depletion of reduced glutathione (GSH), as well as key antioxidant defenses (superoxide dismutase (SOD) and catalase (CAT), was effectively mitigated by the NG and HP. Furthermore, NG and HP suppressed the CIS-triggered inflammation by inhibiting NF-κB activation and downregulating the release of major pro-inflammatory mediators (tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, IL-8, IL-1β). They also counteracted CIS-induced apoptosis by reducing phosphorylation of protein kinase B (Akt), cytochrome c (Cyt c) release, caspase (Cas) activation, and the Bcl-2-associated X protein (Bax)/Bcl-2 ratio. These findings demonstrated that NG and HP could protect ovarian granulosa cells from CIS-induced damage through dual regulation of the Nrf2/heme oxygenase-1 (HO-1) antioxidant and nuclear factor kappa B (NF-κB) inflammatory axis, highlighting their potential as adjuvant therapies to mitigate chemotherapy-induced ovarian toxicity.
proteins
2026-05-13 | Novel anti-Mullerian hormone receptor 2 binding peptide prevents chemotherapy-related ovarian follicle loss in a mouse model.
Cancer treatments can deplete the ovarian follicle reserve causing infertility and early menopause, with subsequent decline in cardiovascular, cognitive, and overall health. Medical measures to prevent this chemotherapy-induced ovarian damage are currently not available. Anti-Müllerian hormone (AMH) is important for preserving the ovarian follicle pool via downregulation of granulosa cell replication and function. Despite proven therapeutic ability to protect the ovarian follicle number in mice exposed to chemotherapy, AMH is not approved for human use. To overcome this gap, we created and patented a peptide designed to specifically bind to the AMH receptor, AMHR2 binding peptide (AMHR2BP), that could serve as an alternative therapeutic treatment. By activating the same downstream signaling and replicating the biological effects of natural AMH, we sought to investigate whether AMHR2BP could protect the follicle pool in both natural and accelerated ovarian aging mouse models. Here, we present a series of in vitro and in vivo translational studies to verify AMHR2BP's affinity, specificity, mechanism of action, stability, and in vivo toxicity and efficacy. We performed immunofluorescence, immunoprecipitation, real-time RT-PCR, mass spectrometry, histological, and immunohistochemistry testing to validate our findings on two levels, gene activation and protein translation. We found that AMHR2BP activates the same SMAD signaling pathways as AMH and ultimately preserves the ovarian follicle pool by preventing the progression of primordial to antral follicles in naturally aging mice. Additionally, we demonstrated that AMHR2BP prevents follicle loss in an accelerated, chemotherapy-induced ovarian aging model, thereby effectively preventing identifiable ovarian damage. Importantly, AMHR2BP also portrays excellent plasma stability with no detectable toxicity. By mimicking the function of AMH, AMHR2BP represents a new medical therapeutic strategy to preserve fertility and reduce long-term reproductive and health risks from chemotherapy treatments.
2025-12-25 | Asymptotic Analysis Applied to Combined TRAIL-Oncolytic Virus and PAC-1 Therapy for Granulosa Cell Tumors of the Ovary
Granulosa cell tumors (GCTs) of the ovary present a therapeutic challenge due to their rarity, recurrence potential, and limited responsiveness to conventional treatments. The mathematical model describes the combination treatment of a TRAIL-producing oncolytic virus with PAC-1, a small-molecule activator of procaspase. The model integrates tumor cell dynamics, viral replication, TRAIL-mediated apoptosis, and PAC-1 pharmacodynamics to simulate treatment responses under various conditions.\The mathematical results suggest that synergistic interaction between the oncolytic virus and PAC-1 can significantly enhance tumor cell killing while minimizing systemic toxicity. These findings offer mechanistic insights and predictive guidance for the preclinical development of this combination therapy, providing a promising strategy for the treatment of GCTs. In this study, we applied a new method to find the base functions to approximate the solution received from the method of directly defining the inverse mapping for systems of nonlinear differential equations to the mathematical model of ovarian cancer.
2025-10-10 | Pancreatic Damage in Ovarian Cancer-Associated Cachexia Is Driven by Activin A Signalling.
Cancer-associated cachexia (CAC) is a severe metabolic disorder characterized by involuntary weight loss, skeletal muscle atrophy and adipose tissue depletion. It is a major contributor to morbidity and mortality in the advanced stages of various cancers. However, the impact of CAC on the pancreas remains largely unexplored. We used mice with constitutively active PI3K in oocytes, generated through a Cre-inducible Pik3ca* knock-in allele driven by Gdf9-icre and performed histological and molecular analyses of the pancreas during cachexia development. Additionally, we examined pancreatic changes following ovariectomy and administration of Follistatin 288 (FST288). Mice that developed cachexia symptoms associated with granulosa cell tumour (GCT) growth exhibited significant pancreatic atrophy compared to controls (Cre+ vs. Cre- at PD83, p < 0.0001), including reduced size of individual acinar cells (102.99 ± 12.19 μm2 vs. 207.94 ± 24.85 μm2 at PD83, p < 0.0001) and acinar units (346.41 ± 169.22 μm2 vs. 1193.59 ± 136.01 μm2 at PD83, p < 0.0001), despite comparable food intake between groups. Acinar cells exhibited a decrease in zymogen granules, reduced amylase expression and diminished amylase activity in both serum (0.29 ± 0.08 vs. 1.41 ± 0.40, p < 0.001) and tissue (0.37 ± 0.14 vs. 1.05 ± 0.29, p < 0.01). In contrast, pancreatic islets remained intact, as evidenced by histological analysis and preserved insulin expression. The pancreas of PD83 Cre+ mice also developed fibrosis and acinar cell death, characterized by elevated expression of collagen IV and α-SMA, and TUNEL-positive signals in acinar cells, respectively. Ovariectomy preserved body weight (2.66 ± 1.30 g for Cre+/OVX vs. 1.60 ± 0.97 g for Cre-) compared to Cre+ mice (-3.66 g) and maintained pancreatic function, suggesting that tumour-derived factors from GCT contribute to the severity of cachexia. Acinar cells showed high expression of ACVR2B, leading to activation of downstream p-SMAD3 signalling. Accordingly, activin A directly induced acinar cell atrophy in both ex vivo cultured pancreas (79.27 ± 19.03 μm2 vs. 171.14 ± 27.01 μm2, p < 0.0001) and 266-6 acinar cells, as evidenced by reduced acinar cell size and decreased amylase production. Injection of FST288, an activin A inhibitor, rescued pancreatic acinar atrophy (252.95 ± 11.59 μm2 in Cre+/FST288 vs. 97.25 ± 12.37 μm2 in Cre+, p < 0.001) without affecting GCT tumour size. Ex vivo culture of pancreas and 266-6 acinar cells exposed to activin A confirmed that activin A directly induces pancreatic damage. These findings demonstrate pancreatic damage occurs during CAC development and highlight the critical role of activin A in this process. Targeting activin A signalling may represent a promising therapeutic strategy to mitigate cachexia in cancer patients and preserve pancreatic function.
2025-04-09 | Hecate-FSHβ33-53C/S lytic peptide conjugate selectively kills targeted follicle stimulating hormone receptor (FSHR)-positive cancer cells.
The follicle stimulating hormone (FSH) receptor (FSHR), is expressed primarily in the gonads, also found in ovarian and prostate cancers, and in tumor vessel endothelial cells. We investigated the potential of a targeted cytotoxic approach using Hecate-FSHβ, a conjugate derived from a lytic peptide Hecate, an analog of bee venom melittin, and the β subunit of FSH, to selectively eliminate FSHR-positive cancer cells. Hecate-FSHβ-mediated cytotoxicity was tested in human granulosa tumor cell line KGN, human embryonic kidney HEK-293 cell line stably transfected with human FSHR cDNA (HEK293-FSHR) and mock-transfected HEK-293 cells as FSHR-negative control cells. Tested variant Hecate-FSHβ33-53C/S with cysteine residues replaced by serine, was evaluated for its cytotoxicity towards FSHR-positive cells. Hecate-FSHβ33-53C/S demonstrated the highest specific cytotoxicity towards FSHR-positive cells (KGN and HEK293-FSHR vs. control). In competition studies, cotreatment with recombinant human FSH (rhFSH) reduced the cytotoxic effect of the conjugate on these cells, highlighting FSHR specificity. In xenograft models of HEK293-FSHR, Hecate-FSHβ33-53C/S alone or in combination with a gonadotropin releasing hormone (GnRH) antagonist (Cetrorelix, CTX) significantly inhibited tumor growth. No synergistic effect was observed with co-administered Hecate-FSHβ33-53C/S and CTX. Hecate-FSHβ33-53C/S induced necrosis in tumor cells, whereas CTX triggered apoptosis. Hecate-FSHβ33-53C/S did not produce any side effects. CTX treatment caused increased spleen size and inhibited spermatogenesis, leading to reduced testis weight, which aligns with expected gonadal effects. Hecate-FSHβ33-53C/S is highly effective in selectively targeting and killing FSHR-expressing cancer cells, with minimal side effects, suggesting its potential as a therapeutic option for cancers expressing FSH receptors.
2025-01-14 | Protective effect of luteinizing hormone on frozen-thawed ovarian follicles and granulosa cells.
Ovarian tissue cryopreservation addresses critical challenges in fertility preservation for prepubertal female cancer patients, such as the lack of viable eggs and hormonal deficiencies. However, mitigating follicle and granulosa cell damage during freeze-thaw cycles remains an urgent issue. Luteinizing hormone (LH), upon binding to luteinizing hormone receptors (LHR) on granulosa cells, enhances estrogen synthesis and secretion, contributing to the growth of granulosa cells and follicles. This study examined mouse ovarian follicles and granulosa cells to identify optimal LH treatments using morphological assessments and LIVE/DEAD assays. The study found significant increases in the expression of Leucine-rich G-protein-coupled receptor 5 (Lgr5) and Forkhead box L2 (Foxl2) in mural and cumulus granulosa cells under LH influence, alongside marked reductions in active caspase-3 expression. Double immunofluorescence of Ki67 with Foxl2 and Lgr5 revealed ongoing proliferative activity in granulosa cells post freeze-thaw. In addition, LH treatment significantly boosted the expression of transforming growth factor (TGF-β) and its superfamily members in both granulosa cells and oocytes. These findings suggest that LH addition during cryopreservation can diminish damage to follicles and granulosa cells, offering new strategies to enhance the efficacy of mammalian ovarian cryopreservation.
antibodies
2026-05-27 | A Narrative Review of Recent Insights on Nerve Growth Factor Signaling in Physiological and Pathological Ovarian Processes in Mammals.
Nerve Growth Factor (NGF), a member of the neurotrophin family, is currently regarded as a key regulator of ovarian physiology beyond its well-known neurotrophic functions. The mammalian ovary is one of the most highly innervated peripheral organs. Increasing evidence indicates that NGF and its receptors, TrkA and p75NTR, are widely expressed in ovarian tissues. Through the activation of the PI3K/AKT, MAPK/ERK, and PLCγ signaling pathways, NGF influences granulosa cell proliferation, steroidogenesis, and ovulation. Physiological levels of NGF are essential for primordial follicle activation, FSH receptor expression, and effective bidirectional communication between oocytes and surrounding somatic cells. As a result, NGF also regulates oocyte maturation and developmental competence. The disruption of NGF signaling can lead to serious health issues. Both low and high levels of NGF negatively affect folliculogenesis and fertility. Elevated intraovarian NGF results in sympathetic over-innervation, altered steroid production, and polycystic ovarian features. In addition, increased NGF expression has been linked to endometriosis and ovarian cancer progression. Clinical studies further suggest that follicular NGF levels may serve as indicators of ovarian reserve and reproductive outcomes in assisted reproduction. This narrative review synthesizes the current knowledge on NGF roles in ovarian physiology and disease. It highlights NGF' dual functions as a central regulator of follicular dynamics, and as a potential biomarker and therapeutic target for common reproductive system diseases.
2026-01-24 | Expression of antibody drug conjugate target antigens in ovarian sex cord stromal Tumors.
The purpose of this study is to determine antibody-drug conjugate (ADC) target antigen expression in ovarian sex cord stromal tumors (SCSTs). This was a retrospective, single-institution study examining SCSTs for immunohistochemical (IHC) expression of antigens targeted by five FDA-approved ADCs (TROP-2, HER2, folate receptor alpha, nectin-4, and tissue factor) plus expression of delta-like ligand 3 (DLL3), the target of tarlatamab-dlle, a bispecific T-cell engager. Demographic and clinical information were abstracted from medical records. Differences in H-scores were evaluated using the Wilcoxon rank-sum tests and correlation between variables was assessed using Spearman's rank correlation coefficient. Thirty-two SCST samples from thirty individual patients were included. The most common diagnosis was adult type granulosa cell tumor. Minimal to no staining was observed for the antigens TROP-2, HER2, folate receptor alpha, nectin-4, and DLL3. Frequent staining for tissue factor was observed in adult GCTs with a median H-score of 49.6 (interquartile range: 1.3-122.8) and 72% had ≥1% expression of tissue factor. A modest positive correlation between H-score and estrogen receptor positivity was noted (Spearman's rho = 0.49, p = 0.06). The tissue factor H-score for the single juvenile GCT was 43.2 and for the two Sertoli Leydig tumors, H-scores were 1 and 10.2. The pervasive absence of HER2, folate receptor alpha, DLL3, TROP-2, and nectin-4 expression in SCSTs suggests ADCs targeting these antigens may be of limited clinical benefit. The design of clinical trials investigating the use of tissue factor-targeting ADCs for the treatment of adult GCTs warrants future consideration.
2025-07-03 | TRIB3 regulates Cx43 expression in human granulosa cells via the Wnt/β-catenin signaling pathway: an in vitro study.
Increased expression of tribbles pseudokinase 3 (TRIB3), which is attributed to increased levels of free fatty acids in the follicular fluid of women with obesity, is correlated with reduced oocyte developmental competence. Connexin 43 (Cx43) is a crucial element of gap junctions in human ovarian granulosa cells (GCs) and facilitates oocyte development. This study aimed to investigate whether high TRIB3 expression and decreased oocyte development potential is regulated by aberrant Cx43 expression in human GCs under high fatty acid culture conditions. GCs from women undergoing in vitro fertilization were grouped by body mass index (BMI). The human granulosa-like tumor cell line (KGN) was exposed to palmitic acid (PA), and its effect on cell apoptosis, viability and proliferation was assessed. The effects of TRIB3 overexpression and knockdown on KGN cells were analyzed. Gene and protein expression, along with the activity of the Wnt/β-catenin signaling pathway, were analyzed using quantitative reverse transcription-PCR, western blotting, immunofluorescence staining, and co-immunoprecipitation. Gap junction function was assessed using a scrape loading/dye transfer assay. Fertilization rates declined in women with overweight or obesity. In the GCs of these women, TRIB3 and Cx43 were upregulated. The protein expression of TRIB3 and Cx43 increased in KGN cells exposed to PA. TRIB3 overexpression led to increased Cx43 expression, whereas TRIB3 knockdown resulted in reduced Cx43 expression. Further mechanistic investigations revealed that TRIB3 upregulated Cx43 by reducing the degradation of β-catenin in the Wnt signaling pathway. TRIB3 overexpression can enhance Cx43 expression in human GCs by inhibiting the Wnt/β-catenin pathway, thereby reducing oocyte maturation rate. These findings could be used to improve the quality of female oocytes and enhance infertility therapy in the clinic.
2024-12-16 | Data from SWOG/NCI Phase II Dual Anti–CTLA-4/PD-1 Blockade in Rare Tumors: Nonepithelial Ovarian Cancer
<div>AbstractPurpose:<p>The role of dual checkpoint inhibition (ipilimumab at 1 mg/kg intravenously every 6 weeks and nivolumab at 240 mg intravenously every 2 weeks) in advanced rare/ultrarare nonepithelial ovarian cancers is yet to be explored.</p>Patients and Methods:<p>Dual anti–CTLA-4 and anti–PD-1 blockade in rare tumor is a prospective, multicenter (1,016 US sites), multicohort, single-arm phase II trial conducted through the Early Therapeutics and Rare Cancer SWOG/NCI Committee, assessing ipilimumab (anti–CTLA-4; 1 mg/kg every 6 weeks) and nivolumab (anti–PD-1; 240 mg every 2 weeks) in adults with advanced nonepithelial ovarian cancers who lack beneficial standard therapy. The primary outcome was overall response rate [ORR; complete response (CR)/partial response (PR)]; secondary outcomes were progression-free survival (PFS), overall survival, clinical benefit rate [stable disease (SD) ≥6 months plus ORR], and toxicity.</p>Results:<p>Seventeen patients (median age: 64; number of prior therapies ranged from 0 to 8 with no immunotherapy exposure; eight granulosa, six carcinosarcomas, one Sertoli–Leydig, one yolk sac, and one Wolffian) were evaluated. In granulosa cell tumors, ORR was 25% (<i>n</i> = 2/8; one CR and one PR) and clinical benefit rate was 50% (<i>n</i> = 4/8); PFS was 58.3 (CR), 50.7+ (PR), 30.4 (SD), and 8.7 (SD) months. Median PFS was 3.5 months [95% confidence interval, 1.7–11.2 months]; median overall survival was 42.5 months (95% confidence interval, 10.1 months–not reached). One Sertoli–Leydig cell tumor showed a 22% regression (PFS, 11.2 months). Carcinosarcomas had no response. Three participants (18%) discontinued treatment due to grade 3 to grade 4 adverse events.</p>Conclusions:<p>Ipilimumab–nivolumab shows activity in treatment-refractory granulosa cell tumors, with 25% (<i>n</i> = 2/8) of patients experiencing either CR or PR lasting more than 4 years.</p></div>
2024-12-09 | Gain-of-Function Chromatin Remodeling Activity of Oncogenic FOXL2C134W Reprograms Glucocorticoid Receptor Occupancy to Drive Granulosa Cell Tumors.
Adult type ovarian granulosa cell tumors (AGCT) are rare malignancies with the near universal c.C402G (p.Cys134Trp) somatic mutation in FOXL2, a forkhead box family transcription factor important for ovarian function. Relapsed AGCT is incurable, but the mechanism of the unique FOXL2 mutation could confer therapeutic vulnerabilities. To identify FOXL2C134W-dependent pharmacologic synergies, we created and characterized endogenous FOXL2 isogenic AGCT cells and an AGCT tumoroid biobank. A drug screen identified that glucocorticoids promote FOXL2C134W-dependent AGCT growth. Epigenetic investigation revealed that the Cys134Trp mutation exposes latent DNA sequence-specific chromatin remodeling activity in FOXL2. FOXL2C134W-dependent chromatin remodeling activity redirected glucocorticoid receptor chromatin occupancy to drive hyaluronan synthase 2 gene expression and increase extracellular hyaluronan secretion. Treatment of AGCT models with hyaluronidase reduced viability, and dexamethasone rescued this effect. Combinatorial drug-drug interaction experiments demonstrated that dexamethasone antagonizes the potency of paclitaxel, a chemotherapy agent frequently used in the treatment of AGCT. Thus, gain-of-function pioneering activity contributes to the oncogenic mechanism of FOXL2C134W and creates a potentially targetable synergy with glucocorticoid signaling. Significance: Glucocorticoids promote granulosa cell tumor growth via epigenetic coregulation with the disease driver FOXL2C134W, providing mechanistic insight into disease oncogenesis and uncovering a potential treatment strategy.
oligonucleotides
2024-05-29 | Paxillin regulates androgen receptor expression associated with granulosa cell focal adhesions.
Paxillin is a ubiquitously expressed adaptor protein integral to focal adhesions, cell motility, and apoptosis. Paxillin has also recently been implicated as a mediator of nongenomic androgen receptor (AR) signaling in prostate cancer and other cells. We sought to investigate the relationship between paxillin and AR in granulosa cells (GCs), where androgen actions, apoptosis, and focal adhesions are of known importance, but where the role of paxillin is understudied. We recently showed that paxillin knockout in mouse GCs increases fertility in older mice. Here, we demonstrate that paxillin knockdown in human granulosa-derived KGN cells, as well as knockout in mouse primary GCs, results in reduced AR protein but not reduced mRNA expression. Further, we find that both AR protein and mRNA half-lives are reduced by approximately one-third in the absence of paxillin, but that cells adapt to chronic loss of paxillin by upregulating AR gene expression. Using co-immunofluorescence and proximity ligation assays, we show that paxillin and AR co-localize at the plasma membrane in GCs in a focal adhesion kinase-dependent way, and that disruption of focal adhesions leads to reduced AR protein level. Our findings suggest that paxillin recruits AR to the GC membrane, where it may be sequestered from proteasomal degradation and poised for nongenomic signaling, as reported in other tissues. To investigate the physiological significance of this in disorders of androgen excess, we tested the effect of GC-specific paxillin knockout in a mouse model of polycystic ovary syndrome (PCOS) induced by chronic postnatal dihydrotestosterone (DHT) exposure. While none of the control mice had estrous cycles, 33% of paxillin knockout mice were cycling, indicating that paxillin deletion may offer partial protection from the negative effects of androgen excess by reducing AR expression. Paxillin-knockout GCs from mice with DHT-induced PCOS also produced more estradiol than GCs from littermate controls. Thus, paxillin may be a novel target in the management of androgen-related disorders in women, such as PCOS.
2024-03-14 | miR-128-3p Regulates Follicular Granulosa Cell Proliferation and Apoptosis by Targeting the Growth Hormone Secretagogue Receptor.
The proliferation and apoptosis of granulosa cells (GCs) affect follicle development and reproductive disorders, with microRNAs playing a crucial regulatory role. Previous studies have shown the differential expression of miR-128-3p at different stages of goat follicle development, which suggests its potential regulatory role in follicle development. In this study, through the Cell Counting Kit-8 assay, the EDU assay, flow cytometry, quantitative real-time polymerase chain reaction, Western blot, and the dual-luciferase reporter assay, we used immortal human ovarian granulosa tumor cell line (KGN) cells as materials to investigate the effects of miR-128-3p and its predicted target gene growth hormone secretagogue receptor (GHSR) on GC proliferation and apoptosis. The results show that overexpression of miR-128-3p inhibited the proliferation of KGN cells, promoted cell apoptosis, and suppressed the expression of proliferating cell nuclear antigen (PCNA) and B-cell lymphoma-2 (BCL2) while promoting that of Bcl-2 associated X protein (BAX). The dual-luciferase reporter assay revealed that miR-128-3p bound to the 3' untranslated region sequence of GHSR, which resulted in the inhibited expression of GHSR protein. Investigation of the effects of GHSR on GC proliferation and apoptosis revealed that GHSR overexpression promoted the expression of PCNA and BCL2, enhanced GC proliferation, and inhibited cell apoptosis, whereas the opposite effects were observed when GHSR expression was inhibited. In addition, miR-128-3p and GHSR can influence the expression of extracellular signal-regulated kinase 1/2 protein. In conclusion, miR-128-3p inhibits KGN cell proliferation and promotes cell apoptosis by downregulating the expression of the GHSR gene.
2023-11-20 | Detection of FOXL2 C134W Mutation Status by a Novel BaseScope In Situ Hybridization Assay is Highly Sensitive and Specific for Adult Granulosa Cell Tumors.
Adult granulosa cell tumors (AGCTs) are a molecularly distinct group of malignant ovarian sex cord-stromal tumors (SCSTs) characterized by a nearly ubiquitous c.402C>G/p.C134W mutation in FOXL2 (hereafter referred to as "C134W"). In some cases, AGCT exhibits marked morphologic overlap with other SCSTs and has an identical immunophenotype, and molecular testing may be necessary to help confirm the diagnosis. However, molecular testing is time consuming, relatively expensive, and unavailable in many pathology laboratories. We describe the development and validation of an in situ hybridization (ISH) custom BaseScope assay for the detection of the FOXL2 C134W mutation. We evaluated 106 ovarian SCSTs, including 78 AGCTs, 9 juvenile granulosa cell tumors, 18 fibromas (cellular and conventional), and 1 SCST, not otherwise specified, as well as 53 epithelial ovarian tumors (42 endometrioid carcinomas and 11 carcinosarcomas) and 1 STK11 adnexal tumor for the presence or absence of FOXL2 wild-type and FOXL2 C134W RNA expression via BaseScope-ISH. Fifty-one tumors had previously undergone DNA sequencing of the FOXL2 gene. Across the entire cohort, the FOXL2 C134W probe staining was positive in 77 of 78 (98.7%) AGCTs. Two of 81 (2.5%) non-AGCTs also showed positive staining, both of which were epithelial ovarian tumors. The assay worked in tissue from blocks >20 years old. There was 100% concordance between the FOXL2 sequencing and BaseScope-ISH results. Overall, assessment of FOXL2 mutation status by custom BaseScope-ISH demonstrated 98.7% sensitivity and 97.5% specificity for the diagnosis of AGCT. BaseScope-ISH for FOXL2 C134W represents a reasonable alternative to sequencing, is quicker and less expensive, and is more easily incorporated than molecular testing into many pathology laboratories. It also has the advantage of requiring less tissue, and the neoplastic cells can be directly visualized on stained sections.
2023-10-31 | SP1 transcriptionally upregulates the expression of APOC3 in KGN cells to promote disease progression by regulating TLR2/NF-κB signalling pathway.
Apolipoprotein C3 (APOC3) is known for its important functions in metabolism-related diseases. However, the function and molecular mechanism of APOC3 in polycystic ovarian syndrome (PCOS) have not been reported. Quantitative polymerase chain reaction and western blot assays were used to detect the expression of APOC3 in KGN cells. Small interference APOC3 (siAPOC3) was applied to reduce APOC3 expression, and the proliferation ability of human granulosa cell line (KGN cells) was measured by cell counting kit-8 and colony formation assays. The protein levels of key genes related to apoptosis were detected by western blot assay. The transcriptional regulator of APOC3 was predicted by the UCSC and PROMO website, and verified by dual luciferase assay. siAPOC3 and pcDNA3.1-specific protein 1 (SP1) vector were co-transfected into KGN cells to detect the function of SP1 and APOC3 in KGN cells. APOC3 was overexpressed in KGN cells, and siAPOC3 transfection significantly reduced the growth ability of KGN cells and increased the apoptosis ability of KGN cells. SP1 directly bound to the promoter of APOC3 and transcriptional regulated APOC3 expression. Overexpression of SP1 increased the growth ability of KGN cells and decreased the apoptosis ability of KGN cells, which were reversed after siAPOC3 transfection. The increased levels of toll-like receptor 2 (TLR2) and p65 phosphorylation (p-P65) nuclear factor kappa B (NF-κB) caused by SP1 overexpression were inhibited by siAPOC3 transfection. APOC3, transcriptionally regulated by SP1, promoted the growth of KGN cells, and inhibited the apoptosis by regulating TLR2/NF-κB signalling pathway.
2023-07-18 | miR-34a-FOXP1 Loop in Ovarian Cancer
Ovarian cancer (OC) is the main cause of gynecological cancer mortality in most developed countries. microRNA (miR) expression dysregulation has been highlighted in human cancers, and miR-34a is found to be downregulated and associated with inhibition of tumor growth and invasion in several malignancies, including OC. The winged helix transcription factor forkhead box P1 (FOXP1) is reported as either an oncogene or tumor suppressor in various cancers. This study aimed to elucidate potential clinical and biological associations of miR-34a and transcription factor FOXP1 in OC. We investigated nine OC patients' blood samples and two OC cell lines (SKOV-3 and OVCAR-3) using quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) to determine both miR-34a and FOXP1 expressions. We have found that miR-34a and FOXP1 are reversely correlated in both in vitro and in vivo. Inhibition of miR-34a transiently led to upregulation of FOXP1 mRNA expression and increased cellular invasion in vitro. Our data indicate that miR-34a could be a potential biomarker for improving the diagnostic efficiency of OC, and miR-34a overexpression may reduce OC pathogenesis by targeting FOXP1.
other
2026-09-01 | Brown and white adipocyte like cells derived exosomes differentially regulate stemness, hormone secretion, apoptosis, and lipid metabolism in model cells.
White and brown adipocytes show therapeutic potential, yet their exosomes mediated cell-specific effects remain poorly understood. This study aimed to assess the impact of conditioned media (CM) or exosomes derived from brown and white adipocyte-like cells (BALCs and WALCs, respectively) on four human cell types, including ovarian granulosa cells (OGCs), MCF-7 breast cancer cells, human adipose-derived stem cells (hADSCs), and human umbilical cord mesenchymal stem cells (hUCMSCs). Isolated hADSCs were differentiated into BALCs and WALCs using specific induction media. Corresponding exosomes were isolated from BALCs or WALCs derived CM and characterized. Human cell types-representing stem, somatic, and cancer cells-were treated with CM or exosomes. Cellular responses and lipidomic profiling were evaluated using molecular biology techniques and mass spectrometry, respectively. BALCs-derived exosomes significantly increased estradiol secretion from ovarian granulosa cells (p < 0.05). In MCF-7 cells, CM had a stronger pro-apoptotic effect than exosomes (>2-fold). In hADSCs, both CM and exosomes promoted osteogenic differentiation (2.3-fold) and reduced stemness markers proteins Oct4 and Sox2 (p < 0.05). When treated with BALCs-derived exosomes, hADSCs displayed the highest number of differentially expressed lipids, primarily upregulated hits. Pathway analysis indicated that BALCs-derived exosomes promoted lipid storage and membrane remodeling in hADSCs, enhanced energy metabolism in granulosa cells, reduced fatty acid oxidation in MCF-7 cells, and decreased lipid storage in hUCMSCs. Our results highlight a potential modulatory role of adipocyte-derived exosomes in intercellular communication. The findings of this study may be applicable in regenerative medicine, metabolic regulation and cancer therapeutic approaches.
2026-05-20 | Knockout of filamin A in KGN granulosa tumor cells impairs proliferation, cell cycle progression, migration, and cytoskeletal organization under mechanical stress
Abstract Background Filamin A (FLNA) is an actin-binding protein that regulates mechanosensitivity and functions as an intracellular signaling scaffold in various cell types. It has also been implicated in tumor growth. We recently reported FLNA expression in human ovarian granulosa cells and in KGN cells, a granulosa cell tumor (GCT) line. Results Immunohistochemistry analysis of 51 GCT samples revealed heterogeneous FLNA expression, with approximately 20% showing weak, 18% strong, and the majority moderate expression. We therefore conducted functional studies in KGN cells using CRISPR/Cas9 gene editing. A proteomic approach revealed marked changes in protein abundance upon FLNA depletion: proteins with increased abundance were predominantly related to adhesion, cytoskeletal organization, regulation of cell shape, and lipid metabolic process, whereas those with decreased abundance were associated with DNA replication, cell division, and cell cycle regulation. FLNA-knockout cells showed enlarged cell sizes, reduced proliferation, and slightly affected steroidogenesis. Disruption of FLNA further reduced migration velocity, altered actin cytoskeletal alignment under flow, and modified expression of genes involved in cytoskeletal architecture, adhesion, and mechanosensing under shear stress. Conclusions Our results identify crucial roles of FLNA in shaping the cellular architecture, motility, and proliferation of KGN cells. Consequently, alterations in FLNA expression may influence intracellular signaling, and responsiveness to mechanical cues in both physiological and pathological contexts.
2025-09-18 | Fertility Preservation in an 18-Year-Old Female with an Ovarian Granulosa Cell Tumor: A Case Report
Objective: Ovarian granulosa cell tumors (GCTs) are rare malignancies that can profoundly affect fertility, particularly in young women. This case report describes a successful fertility preservation strategy in an 18-year-old female diagnosed with a Stage I ovarian granulosa cell tumor. Methods: A multidisciplinary team implemented a fertility preservation protocol involving preoperative ovarian stimulation, oocyte retrieval, and cryopreservation. This was followed by staging laparotomy and unilateral salpingo-oophorectomy. Results: A total of 10 mature oocytes were successfully cryopreserved. Histology confirmed a Stage I granulosa cell tumor with no evidence of metastasis. The patient tolerated adjuvant therapy well and remains disease-free at follow-up. Conclusion: The successful application of controlled ovarian stimulation and oocyte cryopreservation prior to definitive surgery demonstrates the feasibility of proactive fertility preservation in hormonally active tumors when managed with careful monitoring. Furthermore, the patient’s positive postoperative course and continued disease-free status at 18 months reinforce the safety and efficacy of fertility-sparing surgical strategies in Stage I GCTs.
2025-05-03 | Mitigating chemotherapy-induced granulosa cell damage: role of hUCMSC-EVs in regulating the lncRNA HCP5-miR-20a-5p-YAP1 network.
The substantial apprehension facing young cancer patients revolves around the onset of chemotherapy-induced premature ovarian failure (POF), primarily linked to damage inflicted upon granulosa cells (GCs). The inquiry delves into the protective role of extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUCMSCs) in mitigating chemotherapy-induced ovarian failure. Specifically, we investigated the mechanism by which hUCMSC-EVs deliver the long non-coding RNA (lncRNA) HCP5 to regulate DNA damage repair in GCs via the miR-20a-5p/YAP1 axis. The detection of differentially expressed lncRNAs in GC injury resulting from cyclophosphamide (CP) was conducted through transcriptome sequencing. hUCMSC-EVs were isolated, characterized, and co-cultured with CP-injured GCs. Functional assays such as CCK-8, TUNEL, and ELISA were performed to evaluate GC viability, apoptosis, and ovarian endocrine function. Experimental validation of the interactions involving HCP5, miR-20a-5p, and YAP1 was achieved through performing luciferase reporter assays, RNA immunoprecipitation experiments, and Western blot (WB) analyses. HCP5 was significantly enriched in hUCMSC-EVs and effectively delivered into GCs. This resulted in improved GC viability, reduced apoptosis, and enhanced DNA repair. Mechanistically, HCP5 sponged miR-20a-5p, leading to the upregulation of YAP1, which in turn mitigated CP-induced GC damage. In vivo experiments further demonstrated that hUCMSC-EVs prevented CP-induced POF through modulation of the HCP5-miR-20a-5p-YAP1 axis. Our research underscores the therapeutic potential of hUCMSC-EVs in delivering HCP5 to promote DNA repair in GCs, thereby preventing chemotherapy-induced POF. This study provides a novel molecular framework for future therapeutic strategies aimed at protecting ovarian function during chemotherapy.
2025-04-03 | Granulosa Cell Death is a Significant Contributor to DNA-Damaging Chemotherapy-Induced Ovarian Insufficiency.
Typically, DNA-damaging chemotherapy (CTx) regimens have a gonadotoxic effect and cause premature ovarian insufficiency (POI), characterized by infertility and estrogen deficiency. However, whether loss of granulosa cells killed directly by CTx contributes significantly to POI has not been determined. To address this issue, we used a previously established mouse model of CTx-induced POI. The alkylating drugs Busulfan (8.75 mg/kg) and Cyclophosphamide (100 mg/kg) were administered to 8-week-old FVB female mice by intraperitoneal (i.p.) injection three times at 48-hour intervals, after which ovarian tissues were harvested and examined by immunofluorescence. The number of primordial follicles was significantly reduced at day (d)6, whereas the number of growing follicles was relatively unchanged. CTx led to DNA double strand breaks in both oocytes and granulosa cells based on the presence of γH2AX foci. However, markers of apoptosis predominantly labeled granulosa cells in growing follicles. We next examined the effect of inhibiting apoptosis in growing granulosa cells by generating Bak-/-Baxfxfx; Cyp19a1Cre transgenic mice. On d10 after the first CTx, Bak-/-Baxfxfx; Cyp19a1Cre ovaries had fewer apoptotic granulosa cells and more surviving follicles than controls. Furthermore, Bak-/-Baxfxfx; Cyp19a1Cre mice showed better fertility than controls after CTx. Our data suggest that granulosa cell death is a significant contributor to follicle depletion and fertility loss after Cyclophosphamide and Busulfan.
small molecules
2026-09-26 | Treatment Outcomes and Toxicity of Cisplatin-Etoposide-Based Regimens and Carboplatin-Paclitaxel as Adjuvant Chemotherapy for Ovarian Adult Granulosa Cell Tumors: A Multicenter Retrospective Cohort Study.
Background/Objectives: Adult granulosa cell tumors (AGCTs) of the ovary are rare sex cord-stromal malignancies characterized by an indolent clinical course and a persistent risk of late recurrence. Although adjuvant chemotherapy is considered for selected patients with higher-risk disease, the optimal platinum-based regimen remains uncertain because comparative evidence is limited. This study compared disease-free survival (DFS), treatment delivery, and toxicity between bleomycin-etoposide-cisplatin/etoposide-cisplatin (BEP/EP) and carboplatin-paclitaxel (CP) in the postoperative adjuvant setting. Methods: This retrospective, multicenter, nonrandomized comparative cohort study included 58 patients with AGCTs who underwent primary surgery and received adjuvant platinum-based chemotherapy at five tertiary oncology centers in Türkiye between 2010 and 2024. Patients received BEP/EP or CP according to institutional practice and physician discretion. The primary endpoint was disease-free survival (DFS), while secondary endpoints included treatment delivery and treatment-related toxicity. Results: A total of 58 patients were included, with 26 receiving BEP/EP and 32 receiving CP. Patients treated with CP were older, whereas fertility-sparing surgery was more common in the BEP/EP group. After a median follow-up of 75.5 months, 14 recurrences were observed; when deaths without prior recurrence were included according to the prespecified DFS definition, a total of 19 DFS events occurred. Median DFS was 77.2 months in the BEP/EP group and 126.2 months in the CP group. The estimated 5-year DFS rates were 75.5% and 83.5%, respectively, with no statistically significant difference between treatment groups (log-rank p = 0.296). In an exploratory multivariable Cox model restricted to age and treatment regimen, increasing age was associated with a higher risk of a DFS event (adjusted HR 1.046 per year, 95% CI 1.011-1.082; p = 0.010), whereas CP versus BEP/EP was not significantly associated with DFS (adjusted HR 0.426, 95% CI 0.167-1.083; p = 0.073). Treatment completion rates were numerically higher with CP (84.4% vs. 69.2%; p = 0.17). Peripheral neuropathy occurred more frequently with CP (p = 0.007), whereas nephrotoxicity occurred only in the BEP/EP group (p = 0.014). Pulmonary toxicity was observed only with bleomycin-containing treatment, and one patient developed secondary acute myeloid leukemia following BEP. Conclusions: No statistically significant difference in DFS was detected between BEP/EP and CP, while distinct toxicity patterns were observed. Given the retrospective design, limited sample size, and potential for treatment-selection bias and residual confounding, these findings should be considered exploratory and should not be interpreted as evidence of equivalent efficacy between the regimens.
2026-09-22 | Docetaxel for recurrent adult granulosa cell tumor enabling tertiary cytoreduction
Background Adult granulosa cell tumors (AGCTs) are rare ovarian sex cord-stromal tumors characterized by an indolent clinical course, frequent late recurrences, and limited evidence to guide systemic therapy for recurrent disease. Although platinum- and taxane-based regimens have demonstrated activity, optimal treatment after multiple recurrences remains undefined. Case We report a 53-year-old woman with recurrent AGCT who had progressed on multiple chemotherapy and hormonal regimens over eight years. Despite prior exposure to and progression on paclitaxel, her disease responded to single-agent docetaxel for 36 cycles, reducing disease burden sufficiently to enable tertiary cytoreductive surgery. The treatment was well tolerated. Conclusion This case highlights the potential activity of docetaxel in recurrent AGCT and supports its consideration as an alternative taxane, particularly for patients who are not candidates for or have progressed after other systemic therapies including paclitaxel. Reports describing the use of docetaxel in AGCT are limited, with only isolated cases published to date. Given the rarity of AGCT, individual case reports remain a valuable source of evidence to inform treatment decisions. Prospective studies are needed to better define the role of docetaxel in recurrent AGCT.
2026-08-27 | Isorhamnetin and Female Reproduction: Effects on Viability, Hormone Secretion, and Growth Factors.
Background/Objectives: Isorhamnetin is a naturally occurring flavonoid with reported antioxidant, anti-inflammatory, and anticancer properties. Although its biological activities have been extensively investigated, its effects on ovarian cell physiology remain insufficiently characterized. This study aimed to evaluate the influence of isorhamnetin on the viability, steroid hormone secretion, apoptosis, and growth factor signalling in human ovarian cell lines representing both non-tumour and tumour phenotypes. Methods: Human granulosa (HGL5), granulosa tumour (COV434), and epithelial ovarian carcinoma (OVCAR-3) cell lines were treated with isorhamnetin at concentrations of 5-80 μg/mL for 24 h. Cell viability was determined using the AlamarBlueTM assay. The secretion of progesterone, 17β-estradiol, and the presence of transforming growth factor β2 (TGF-β2), transforming growth factor β receptor 2 (TGFBR2), and apoptosis-inducing factor (AIF) was quantified by enzyme-linked immunosorbent assay (ELISA). Results: Isorhamnetin significantly reduced cell viability in a dose-dependent manner, with tumour cell lines exhibiting greater sensitivity than non-tumour granulosa cells. A significant decrease in viability was observed in OVCAR-3 cells from 10 μg/mL onward, whereas COV434 cells showed significant reductions at concentrations of 20 μg/mL and higher. In contrast, HGL5 cell viability was significantly affected only at the highest concentration (80 μg/mL). No significant changes were detected in the secretion of progesterone, 17β-estradiol, TGF-β2, or TGFBR2 in any of the examined cell lines. A significant reduction in AIF production was observed only in HGL5 cells treated with 80 μg/mL isorhamnetin. Conclusions: Isorhamnetin preferentially decreased the viability of ovarian tumour cells while exerting only limited effects on non-tumour granulosa cells, indicating preferential cytotoxic activity toward malignant ovarian cells. The absence of significant changes in steroid hormone production and TGF-β suggests that its antiproliferative effects involve molecular pathways that were not investigated in the present study. These findings support further investigation of isorhamnetin as a potential natural compound for ovarian cancer prevention or adjunctive therapy.
2026-08-11 | A phase II study of androgen receptor inhibition by darolutamide in combination with leuprolide acetate and exemestane in recurrent adult-type ovarian granulosa cell tumor.
To determine the efficacy and safety of darolutamide, exemestane, and leuprolide acetate in recurrent adult-type ovarian granulosa cell tumor (AGCT). This single-arm Phase II cooperative group study included patients with recurrent AGCT who had progressed on a prior aromatase inhibitor. The treatment regimen consisted of darolutamide 600 mg by mouth twice daily, exemestane 25 mg by mouth once daily, and leuprolide acetate 7.5 mg by intramuscular injection every four weeks. Patient accrual occurred from January 2024 to April 2024 with sample size determined using Simon's Optimal two-stage design. The primary measure of efficacy was objective response rate (ORR) measured by Response Evaluation Criteria in Solid Tumors 1.1. Secondary objectives included duration of response, progression-free survival (PFS), overall survival (OS), and safety of the treatment regimen. Seventeen patients were enrolled to complete target accrual of the first stage. Of the 16 evaluable patients, there was one partial response (ORR = 6.25%). The trial did not continue to the second stage as it did not meet its prespecified endpoint. Ten patients (62.5%) were confirmed to have stable disease and five patients (31.25%) were confirmed to have progressive disease. Median PFS was 8.5 months (95% CI: 3.1 months - 12.0 months). Median OS was not reached. There were no grade 4 or grade 5 adverse events attributable to the treatment regimen. Though the trial did not meet its primary endpoint, there was one partial response to the treatment regimen. Clinically meaningful PFS was demonstrated. Clinical Trial Registration Number NCT06169124.
2026-08-02 | Citrus flavonoids mitigate the cisplatin-induced ovarian toxicity via dual modulation of Nrf2/HO-1 pathway and NF-κB axis.
Cisplatin (CIS) is an effective chemotherapeutic agent whose clinical use is limited by off-target toxicity, including damage to ovarian granulosa cells, characterized by oxidative stress and inflammation. Here, the cytoprotective potential of the citrus flavonoids, naringin (NG) and hesperidin (HP), was investigated in ovarian granulosa cells exposed to CIS-induced toxicity. Molecular docking demonstrated that NG and HP exhibited strong binding affinities for key antioxidant, nuclear factor erythroid 2-related factor 2 (Nrf2) and catalase (CAT) as well as anti-apoptotic B-cell lymphoma-2 (Bcl-2) proteins, while CIS preferentially interacted with pro-apoptotic targets. In vitro, CIS dose-dependently reduced cell viability and hormone secretion (progesterone and estradiol), and these reductions were significantly restored by co-treatment with NG or HP. CIS-induced oxidative injury, marked by elevated accumulation of reactive oxygen species (ROS), lipid peroxidation, and depletion of reduced glutathione (GSH), as well as key antioxidant defenses (superoxide dismutase (SOD) and catalase (CAT), was effectively mitigated by the NG and HP. Furthermore, NG and HP suppressed the CIS-triggered inflammation by inhibiting NF-κB activation and downregulating the release of major pro-inflammatory mediators (tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, IL-8, IL-1β). They also counteracted CIS-induced apoptosis by reducing phosphorylation of protein kinase B (Akt), cytochrome c (Cyt c) release, caspase (Cas) activation, and the Bcl-2-associated X protein (Bax)/Bcl-2 ratio. These findings demonstrated that NG and HP could protect ovarian granulosa cells from CIS-induced damage through dual regulation of the Nrf2/heme oxygenase-1 (HO-1) antioxidant and nuclear factor kappa B (NF-κB) inflammatory axis, highlighting their potential as adjuvant therapies to mitigate chemotherapy-induced ovarian toxicity.
proteins
2026-05-13 | Novel anti-Mullerian hormone receptor 2 binding peptide prevents chemotherapy-related ovarian follicle loss in a mouse model.
Cancer treatments can deplete the ovarian follicle reserve causing infertility and early menopause, with subsequent decline in cardiovascular, cognitive, and overall health. Medical measures to prevent this chemotherapy-induced ovarian damage are currently not available. Anti-Müllerian hormone (AMH) is important for preserving the ovarian follicle pool via downregulation of granulosa cell replication and function. Despite proven therapeutic ability to protect the ovarian follicle number in mice exposed to chemotherapy, AMH is not approved for human use. To overcome this gap, we created and patented a peptide designed to specifically bind to the AMH receptor, AMHR2 binding peptide (AMHR2BP), that could serve as an alternative therapeutic treatment. By activating the same downstream signaling and replicating the biological effects of natural AMH, we sought to investigate whether AMHR2BP could protect the follicle pool in both natural and accelerated ovarian aging mouse models. Here, we present a series of in vitro and in vivo translational studies to verify AMHR2BP's affinity, specificity, mechanism of action, stability, and in vivo toxicity and efficacy. We performed immunofluorescence, immunoprecipitation, real-time RT-PCR, mass spectrometry, histological, and immunohistochemistry testing to validate our findings on two levels, gene activation and protein translation. We found that AMHR2BP activates the same SMAD signaling pathways as AMH and ultimately preserves the ovarian follicle pool by preventing the progression of primordial to antral follicles in naturally aging mice. Additionally, we demonstrated that AMHR2BP prevents follicle loss in an accelerated, chemotherapy-induced ovarian aging model, thereby effectively preventing identifiable ovarian damage. Importantly, AMHR2BP also portrays excellent plasma stability with no detectable toxicity. By mimicking the function of AMH, AMHR2BP represents a new medical therapeutic strategy to preserve fertility and reduce long-term reproductive and health risks from chemotherapy treatments.
2025-12-25 | Asymptotic Analysis Applied to Combined TRAIL-Oncolytic Virus and PAC-1 Therapy for Granulosa Cell Tumors of the Ovary
Granulosa cell tumors (GCTs) of the ovary present a therapeutic challenge due to their rarity, recurrence potential, and limited responsiveness to conventional treatments. The mathematical model describes the combination treatment of a TRAIL-producing oncolytic virus with PAC-1, a small-molecule activator of procaspase. The model integrates tumor cell dynamics, viral replication, TRAIL-mediated apoptosis, and PAC-1 pharmacodynamics to simulate treatment responses under various conditions.\The mathematical results suggest that synergistic interaction between the oncolytic virus and PAC-1 can significantly enhance tumor cell killing while minimizing systemic toxicity. These findings offer mechanistic insights and predictive guidance for the preclinical development of this combination therapy, providing a promising strategy for the treatment of GCTs. In this study, we applied a new method to find the base functions to approximate the solution received from the method of directly defining the inverse mapping for systems of nonlinear differential equations to the mathematical model of ovarian cancer.
2025-10-10 | Pancreatic Damage in Ovarian Cancer-Associated Cachexia Is Driven by Activin A Signalling.
Cancer-associated cachexia (CAC) is a severe metabolic disorder characterized by involuntary weight loss, skeletal muscle atrophy and adipose tissue depletion. It is a major contributor to morbidity and mortality in the advanced stages of various cancers. However, the impact of CAC on the pancreas remains largely unexplored. We used mice with constitutively active PI3K in oocytes, generated through a Cre-inducible Pik3ca* knock-in allele driven by Gdf9-icre and performed histological and molecular analyses of the pancreas during cachexia development. Additionally, we examined pancreatic changes following ovariectomy and administration of Follistatin 288 (FST288). Mice that developed cachexia symptoms associated with granulosa cell tumour (GCT) growth exhibited significant pancreatic atrophy compared to controls (Cre+ vs. Cre- at PD83, p < 0.0001), including reduced size of individual acinar cells (102.99 ± 12.19 μm2 vs. 207.94 ± 24.85 μm2 at PD83, p < 0.0001) and acinar units (346.41 ± 169.22 μm2 vs. 1193.59 ± 136.01 μm2 at PD83, p < 0.0001), despite comparable food intake between groups. Acinar cells exhibited a decrease in zymogen granules, reduced amylase expression and diminished amylase activity in both serum (0.29 ± 0.08 vs. 1.41 ± 0.40, p < 0.001) and tissue (0.37 ± 0.14 vs. 1.05 ± 0.29, p < 0.01). In contrast, pancreatic islets remained intact, as evidenced by histological analysis and preserved insulin expression. The pancreas of PD83 Cre+ mice also developed fibrosis and acinar cell death, characterized by elevated expression of collagen IV and α-SMA, and TUNEL-positive signals in acinar cells, respectively. Ovariectomy preserved body weight (2.66 ± 1.30 g for Cre+/OVX vs. 1.60 ± 0.97 g for Cre-) compared to Cre+ mice (-3.66 g) and maintained pancreatic function, suggesting that tumour-derived factors from GCT contribute to the severity of cachexia. Acinar cells showed high expression of ACVR2B, leading to activation of downstream p-SMAD3 signalling. Accordingly, activin A directly induced acinar cell atrophy in both ex vivo cultured pancreas (79.27 ± 19.03 μm2 vs. 171.14 ± 27.01 μm2, p < 0.0001) and 266-6 acinar cells, as evidenced by reduced acinar cell size and decreased amylase production. Injection of FST288, an activin A inhibitor, rescued pancreatic acinar atrophy (252.95 ± 11.59 μm2 in Cre+/FST288 vs. 97.25 ± 12.37 μm2 in Cre+, p < 0.001) without affecting GCT tumour size. Ex vivo culture of pancreas and 266-6 acinar cells exposed to activin A confirmed that activin A directly induces pancreatic damage. These findings demonstrate pancreatic damage occurs during CAC development and highlight the critical role of activin A in this process. Targeting activin A signalling may represent a promising therapeutic strategy to mitigate cachexia in cancer patients and preserve pancreatic function.
2025-04-09 | Hecate-FSHβ33-53C/S lytic peptide conjugate selectively kills targeted follicle stimulating hormone receptor (FSHR)-positive cancer cells.
The follicle stimulating hormone (FSH) receptor (FSHR), is expressed primarily in the gonads, also found in ovarian and prostate cancers, and in tumor vessel endothelial cells. We investigated the potential of a targeted cytotoxic approach using Hecate-FSHβ, a conjugate derived from a lytic peptide Hecate, an analog of bee venom melittin, and the β subunit of FSH, to selectively eliminate FSHR-positive cancer cells. Hecate-FSHβ-mediated cytotoxicity was tested in human granulosa tumor cell line KGN, human embryonic kidney HEK-293 cell line stably transfected with human FSHR cDNA (HEK293-FSHR) and mock-transfected HEK-293 cells as FSHR-negative control cells. Tested variant Hecate-FSHβ33-53C/S with cysteine residues replaced by serine, was evaluated for its cytotoxicity towards FSHR-positive cells. Hecate-FSHβ33-53C/S demonstrated the highest specific cytotoxicity towards FSHR-positive cells (KGN and HEK293-FSHR vs. control). In competition studies, cotreatment with recombinant human FSH (rhFSH) reduced the cytotoxic effect of the conjugate on these cells, highlighting FSHR specificity. In xenograft models of HEK293-FSHR, Hecate-FSHβ33-53C/S alone or in combination with a gonadotropin releasing hormone (GnRH) antagonist (Cetrorelix, CTX) significantly inhibited tumor growth. No synergistic effect was observed with co-administered Hecate-FSHβ33-53C/S and CTX. Hecate-FSHβ33-53C/S induced necrosis in tumor cells, whereas CTX triggered apoptosis. Hecate-FSHβ33-53C/S did not produce any side effects. CTX treatment caused increased spleen size and inhibited spermatogenesis, leading to reduced testis weight, which aligns with expected gonadal effects. Hecate-FSHβ33-53C/S is highly effective in selectively targeting and killing FSHR-expressing cancer cells, with minimal side effects, suggesting its potential as a therapeutic option for cancers expressing FSH receptors.
2025-01-14 | Protective effect of luteinizing hormone on frozen-thawed ovarian follicles and granulosa cells.
Ovarian tissue cryopreservation addresses critical challenges in fertility preservation for prepubertal female cancer patients, such as the lack of viable eggs and hormonal deficiencies. However, mitigating follicle and granulosa cell damage during freeze-thaw cycles remains an urgent issue. Luteinizing hormone (LH), upon binding to luteinizing hormone receptors (LHR) on granulosa cells, enhances estrogen synthesis and secretion, contributing to the growth of granulosa cells and follicles. This study examined mouse ovarian follicles and granulosa cells to identify optimal LH treatments using morphological assessments and LIVE/DEAD assays. The study found significant increases in the expression of Leucine-rich G-protein-coupled receptor 5 (Lgr5) and Forkhead box L2 (Foxl2) in mural and cumulus granulosa cells under LH influence, alongside marked reductions in active caspase-3 expression. Double immunofluorescence of Ki67 with Foxl2 and Lgr5 revealed ongoing proliferative activity in granulosa cells post freeze-thaw. In addition, LH treatment significantly boosted the expression of transforming growth factor (TGF-β) and its superfamily members in both granulosa cells and oocytes. These findings suggest that LH addition during cryopreservation can diminish damage to follicles and granulosa cells, offering new strategies to enhance the efficacy of mammalian ovarian cryopreservation.
antibodies
2026-05-27 | A Narrative Review of Recent Insights on Nerve Growth Factor Signaling in Physiological and Pathological Ovarian Processes in Mammals.
Nerve Growth Factor (NGF), a member of the neurotrophin family, is currently regarded as a key regulator of ovarian physiology beyond its well-known neurotrophic functions. The mammalian ovary is one of the most highly innervated peripheral organs. Increasing evidence indicates that NGF and its receptors, TrkA and p75NTR, are widely expressed in ovarian tissues. Through the activation of the PI3K/AKT, MAPK/ERK, and PLCγ signaling pathways, NGF influences granulosa cell proliferation, steroidogenesis, and ovulation. Physiological levels of NGF are essential for primordial follicle activation, FSH receptor expression, and effective bidirectional communication between oocytes and surrounding somatic cells. As a result, NGF also regulates oocyte maturation and developmental competence. The disruption of NGF signaling can lead to serious health issues. Both low and high levels of NGF negatively affect folliculogenesis and fertility. Elevated intraovarian NGF results in sympathetic over-innervation, altered steroid production, and polycystic ovarian features. In addition, increased NGF expression has been linked to endometriosis and ovarian cancer progression. Clinical studies further suggest that follicular NGF levels may serve as indicators of ovarian reserve and reproductive outcomes in assisted reproduction. This narrative review synthesizes the current knowledge on NGF roles in ovarian physiology and disease. It highlights NGF' dual functions as a central regulator of follicular dynamics, and as a potential biomarker and therapeutic target for common reproductive system diseases.
2026-01-24 | Expression of antibody drug conjugate target antigens in ovarian sex cord stromal Tumors.
The purpose of this study is to determine antibody-drug conjugate (ADC) target antigen expression in ovarian sex cord stromal tumors (SCSTs). This was a retrospective, single-institution study examining SCSTs for immunohistochemical (IHC) expression of antigens targeted by five FDA-approved ADCs (TROP-2, HER2, folate receptor alpha, nectin-4, and tissue factor) plus expression of delta-like ligand 3 (DLL3), the target of tarlatamab-dlle, a bispecific T-cell engager. Demographic and clinical information were abstracted from medical records. Differences in H-scores were evaluated using the Wilcoxon rank-sum tests and correlation between variables was assessed using Spearman's rank correlation coefficient. Thirty-two SCST samples from thirty individual patients were included. The most common diagnosis was adult type granulosa cell tumor. Minimal to no staining was observed for the antigens TROP-2, HER2, folate receptor alpha, nectin-4, and DLL3. Frequent staining for tissue factor was observed in adult GCTs with a median H-score of 49.6 (interquartile range: 1.3-122.8) and 72% had ≥1% expression of tissue factor. A modest positive correlation between H-score and estrogen receptor positivity was noted (Spearman's rho = 0.49, p = 0.06). The tissue factor H-score for the single juvenile GCT was 43.2 and for the two Sertoli Leydig tumors, H-scores were 1 and 10.2. The pervasive absence of HER2, folate receptor alpha, DLL3, TROP-2, and nectin-4 expression in SCSTs suggests ADCs targeting these antigens may be of limited clinical benefit. The design of clinical trials investigating the use of tissue factor-targeting ADCs for the treatment of adult GCTs warrants future consideration.
2025-07-03 | TRIB3 regulates Cx43 expression in human granulosa cells via the Wnt/β-catenin signaling pathway: an in vitro study.
Increased expression of tribbles pseudokinase 3 (TRIB3), which is attributed to increased levels of free fatty acids in the follicular fluid of women with obesity, is correlated with reduced oocyte developmental competence. Connexin 43 (Cx43) is a crucial element of gap junctions in human ovarian granulosa cells (GCs) and facilitates oocyte development. This study aimed to investigate whether high TRIB3 expression and decreased oocyte development potential is regulated by aberrant Cx43 expression in human GCs under high fatty acid culture conditions. GCs from women undergoing in vitro fertilization were grouped by body mass index (BMI). The human granulosa-like tumor cell line (KGN) was exposed to palmitic acid (PA), and its effect on cell apoptosis, viability and proliferation was assessed. The effects of TRIB3 overexpression and knockdown on KGN cells were analyzed. Gene and protein expression, along with the activity of the Wnt/β-catenin signaling pathway, were analyzed using quantitative reverse transcription-PCR, western blotting, immunofluorescence staining, and co-immunoprecipitation. Gap junction function was assessed using a scrape loading/dye transfer assay. Fertilization rates declined in women with overweight or obesity. In the GCs of these women, TRIB3 and Cx43 were upregulated. The protein expression of TRIB3 and Cx43 increased in KGN cells exposed to PA. TRIB3 overexpression led to increased Cx43 expression, whereas TRIB3 knockdown resulted in reduced Cx43 expression. Further mechanistic investigations revealed that TRIB3 upregulated Cx43 by reducing the degradation of β-catenin in the Wnt signaling pathway. TRIB3 overexpression can enhance Cx43 expression in human GCs by inhibiting the Wnt/β-catenin pathway, thereby reducing oocyte maturation rate. These findings could be used to improve the quality of female oocytes and enhance infertility therapy in the clinic.
2024-12-16 | Data from SWOG/NCI Phase II Dual Anti–CTLA-4/PD-1 Blockade in Rare Tumors: Nonepithelial Ovarian Cancer
<div>AbstractPurpose:<p>The role of dual checkpoint inhibition (ipilimumab at 1 mg/kg intravenously every 6 weeks and nivolumab at 240 mg intravenously every 2 weeks) in advanced rare/ultrarare nonepithelial ovarian cancers is yet to be explored.</p>Patients and Methods:<p>Dual anti–CTLA-4 and anti–PD-1 blockade in rare tumor is a prospective, multicenter (1,016 US sites), multicohort, single-arm phase II trial conducted through the Early Therapeutics and Rare Cancer SWOG/NCI Committee, assessing ipilimumab (anti–CTLA-4; 1 mg/kg every 6 weeks) and nivolumab (anti–PD-1; 240 mg every 2 weeks) in adults with advanced nonepithelial ovarian cancers who lack beneficial standard therapy. The primary outcome was overall response rate [ORR; complete response (CR)/partial response (PR)]; secondary outcomes were progression-free survival (PFS), overall survival, clinical benefit rate [stable disease (SD) ≥6 months plus ORR], and toxicity.</p>Results:<p>Seventeen patients (median age: 64; number of prior therapies ranged from 0 to 8 with no immunotherapy exposure; eight granulosa, six carcinosarcomas, one Sertoli–Leydig, one yolk sac, and one Wolffian) were evaluated. In granulosa cell tumors, ORR was 25% (<i>n</i> = 2/8; one CR and one PR) and clinical benefit rate was 50% (<i>n</i> = 4/8); PFS was 58.3 (CR), 50.7+ (PR), 30.4 (SD), and 8.7 (SD) months. Median PFS was 3.5 months [95% confidence interval, 1.7–11.2 months]; median overall survival was 42.5 months (95% confidence interval, 10.1 months–not reached). One Sertoli–Leydig cell tumor showed a 22% regression (PFS, 11.2 months). Carcinosarcomas had no response. Three participants (18%) discontinued treatment due to grade 3 to grade 4 adverse events.</p>Conclusions:<p>Ipilimumab–nivolumab shows activity in treatment-refractory granulosa cell tumors, with 25% (<i>n</i> = 2/8) of patients experiencing either CR or PR lasting more than 4 years.</p></div>
2024-12-09 | Gain-of-Function Chromatin Remodeling Activity of Oncogenic FOXL2C134W Reprograms Glucocorticoid Receptor Occupancy to Drive Granulosa Cell Tumors.
Adult type ovarian granulosa cell tumors (AGCT) are rare malignancies with the near universal c.C402G (p.Cys134Trp) somatic mutation in FOXL2, a forkhead box family transcription factor important for ovarian function. Relapsed AGCT is incurable, but the mechanism of the unique FOXL2 mutation could confer therapeutic vulnerabilities. To identify FOXL2C134W-dependent pharmacologic synergies, we created and characterized endogenous FOXL2 isogenic AGCT cells and an AGCT tumoroid biobank. A drug screen identified that glucocorticoids promote FOXL2C134W-dependent AGCT growth. Epigenetic investigation revealed that the Cys134Trp mutation exposes latent DNA sequence-specific chromatin remodeling activity in FOXL2. FOXL2C134W-dependent chromatin remodeling activity redirected glucocorticoid receptor chromatin occupancy to drive hyaluronan synthase 2 gene expression and increase extracellular hyaluronan secretion. Treatment of AGCT models with hyaluronidase reduced viability, and dexamethasone rescued this effect. Combinatorial drug-drug interaction experiments demonstrated that dexamethasone antagonizes the potency of paclitaxel, a chemotherapy agent frequently used in the treatment of AGCT. Thus, gain-of-function pioneering activity contributes to the oncogenic mechanism of FOXL2C134W and creates a potentially targetable synergy with glucocorticoid signaling. Significance: Glucocorticoids promote granulosa cell tumor growth via epigenetic coregulation with the disease driver FOXL2C134W, providing mechanistic insight into disease oncogenesis and uncovering a potential treatment strategy.
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2024-05-29 | Paxillin regulates androgen receptor expression associated with granulosa cell focal adhesions.
Paxillin is a ubiquitously expressed adaptor protein integral to focal adhesions, cell motility, and apoptosis. Paxillin has also recently been implicated as a mediator of nongenomic androgen receptor (AR) signaling in prostate cancer and other cells. We sought to investigate the relationship between paxillin and AR in granulosa cells (GCs), where androgen actions, apoptosis, and focal adhesions are of known importance, but where the role of paxillin is understudied. We recently showed that paxillin knockout in mouse GCs increases fertility in older mice. Here, we demonstrate that paxillin knockdown in human granulosa-derived KGN cells, as well as knockout in mouse primary GCs, results in reduced AR protein but not reduced mRNA expression. Further, we find that both AR protein and mRNA half-lives are reduced by approximately one-third in the absence of paxillin, but that cells adapt to chronic loss of paxillin by upregulating AR gene expression. Using co-immunofluorescence and proximity ligation assays, we show that paxillin and AR co-localize at the plasma membrane in GCs in a focal adhesion kinase-dependent way, and that disruption of focal adhesions leads to reduced AR protein level. Our findings suggest that paxillin recruits AR to the GC membrane, where it may be sequestered from proteasomal degradation and poised for nongenomic signaling, as reported in other tissues. To investigate the physiological significance of this in disorders of androgen excess, we tested the effect of GC-specific paxillin knockout in a mouse model of polycystic ovary syndrome (PCOS) induced by chronic postnatal dihydrotestosterone (DHT) exposure. While none of the control mice had estrous cycles, 33% of paxillin knockout mice were cycling, indicating that paxillin deletion may offer partial protection from the negative effects of androgen excess by reducing AR expression. Paxillin-knockout GCs from mice with DHT-induced PCOS also produced more estradiol than GCs from littermate controls. Thus, paxillin may be a novel target in the management of androgen-related disorders in women, such as PCOS.
2024-03-14 | miR-128-3p Regulates Follicular Granulosa Cell Proliferation and Apoptosis by Targeting the Growth Hormone Secretagogue Receptor.
The proliferation and apoptosis of granulosa cells (GCs) affect follicle development and reproductive disorders, with microRNAs playing a crucial regulatory role. Previous studies have shown the differential expression of miR-128-3p at different stages of goat follicle development, which suggests its potential regulatory role in follicle development. In this study, through the Cell Counting Kit-8 assay, the EDU assay, flow cytometry, quantitative real-time polymerase chain reaction, Western blot, and the dual-luciferase reporter assay, we used immortal human ovarian granulosa tumor cell line (KGN) cells as materials to investigate the effects of miR-128-3p and its predicted target gene growth hormone secretagogue receptor (GHSR) on GC proliferation and apoptosis. The results show that overexpression of miR-128-3p inhibited the proliferation of KGN cells, promoted cell apoptosis, and suppressed the expression of proliferating cell nuclear antigen (PCNA) and B-cell lymphoma-2 (BCL2) while promoting that of Bcl-2 associated X protein (BAX). The dual-luciferase reporter assay revealed that miR-128-3p bound to the 3' untranslated region sequence of GHSR, which resulted in the inhibited expression of GHSR protein. Investigation of the effects of GHSR on GC proliferation and apoptosis revealed that GHSR overexpression promoted the expression of PCNA and BCL2, enhanced GC proliferation, and inhibited cell apoptosis, whereas the opposite effects were observed when GHSR expression was inhibited. In addition, miR-128-3p and GHSR can influence the expression of extracellular signal-regulated kinase 1/2 protein. In conclusion, miR-128-3p inhibits KGN cell proliferation and promotes cell apoptosis by downregulating the expression of the GHSR gene.
2023-11-20 | Detection of FOXL2 C134W Mutation Status by a Novel BaseScope In Situ Hybridization Assay is Highly Sensitive and Specific for Adult Granulosa Cell Tumors.
Adult granulosa cell tumors (AGCTs) are a molecularly distinct group of malignant ovarian sex cord-stromal tumors (SCSTs) characterized by a nearly ubiquitous c.402C>G/p.C134W mutation in FOXL2 (hereafter referred to as "C134W"). In some cases, AGCT exhibits marked morphologic overlap with other SCSTs and has an identical immunophenotype, and molecular testing may be necessary to help confirm the diagnosis. However, molecular testing is time consuming, relatively expensive, and unavailable in many pathology laboratories. We describe the development and validation of an in situ hybridization (ISH) custom BaseScope assay for the detection of the FOXL2 C134W mutation. We evaluated 106 ovarian SCSTs, including 78 AGCTs, 9 juvenile granulosa cell tumors, 18 fibromas (cellular and conventional), and 1 SCST, not otherwise specified, as well as 53 epithelial ovarian tumors (42 endometrioid carcinomas and 11 carcinosarcomas) and 1 STK11 adnexal tumor for the presence or absence of FOXL2 wild-type and FOXL2 C134W RNA expression via BaseScope-ISH. Fifty-one tumors had previously undergone DNA sequencing of the FOXL2 gene. Across the entire cohort, the FOXL2 C134W probe staining was positive in 77 of 78 (98.7%) AGCTs. Two of 81 (2.5%) non-AGCTs also showed positive staining, both of which were epithelial ovarian tumors. The assay worked in tissue from blocks >20 years old. There was 100% concordance between the FOXL2 sequencing and BaseScope-ISH results. Overall, assessment of FOXL2 mutation status by custom BaseScope-ISH demonstrated 98.7% sensitivity and 97.5% specificity for the diagnosis of AGCT. BaseScope-ISH for FOXL2 C134W represents a reasonable alternative to sequencing, is quicker and less expensive, and is more easily incorporated than molecular testing into many pathology laboratories. It also has the advantage of requiring less tissue, and the neoplastic cells can be directly visualized on stained sections.
2023-10-31 | SP1 transcriptionally upregulates the expression of APOC3 in KGN cells to promote disease progression by regulating TLR2/NF-κB signalling pathway.
Apolipoprotein C3 (APOC3) is known for its important functions in metabolism-related diseases. However, the function and molecular mechanism of APOC3 in polycystic ovarian syndrome (PCOS) have not been reported. Quantitative polymerase chain reaction and western blot assays were used to detect the expression of APOC3 in KGN cells. Small interference APOC3 (siAPOC3) was applied to reduce APOC3 expression, and the proliferation ability of human granulosa cell line (KGN cells) was measured by cell counting kit-8 and colony formation assays. The protein levels of key genes related to apoptosis were detected by western blot assay. The transcriptional regulator of APOC3 was predicted by the UCSC and PROMO website, and verified by dual luciferase assay. siAPOC3 and pcDNA3.1-specific protein 1 (SP1) vector were co-transfected into KGN cells to detect the function of SP1 and APOC3 in KGN cells. APOC3 was overexpressed in KGN cells, and siAPOC3 transfection significantly reduced the growth ability of KGN cells and increased the apoptosis ability of KGN cells. SP1 directly bound to the promoter of APOC3 and transcriptional regulated APOC3 expression. Overexpression of SP1 increased the growth ability of KGN cells and decreased the apoptosis ability of KGN cells, which were reversed after siAPOC3 transfection. The increased levels of toll-like receptor 2 (TLR2) and p65 phosphorylation (p-P65) nuclear factor kappa B (NF-κB) caused by SP1 overexpression were inhibited by siAPOC3 transfection. APOC3, transcriptionally regulated by SP1, promoted the growth of KGN cells, and inhibited the apoptosis by regulating TLR2/NF-κB signalling pathway.
2023-07-18 | miR-34a-FOXP1 Loop in Ovarian Cancer
Ovarian cancer (OC) is the main cause of gynecological cancer mortality in most developed countries. microRNA (miR) expression dysregulation has been highlighted in human cancers, and miR-34a is found to be downregulated and associated with inhibition of tumor growth and invasion in several malignancies, including OC. The winged helix transcription factor forkhead box P1 (FOXP1) is reported as either an oncogene or tumor suppressor in various cancers. This study aimed to elucidate potential clinical and biological associations of miR-34a and transcription factor FOXP1 in OC. We investigated nine OC patients' blood samples and two OC cell lines (SKOV-3 and OVCAR-3) using quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) to determine both miR-34a and FOXP1 expressions. We have found that miR-34a and FOXP1 are reversely correlated in both in vitro and in vivo. Inhibition of miR-34a transiently led to upregulation of FOXP1 mRNA expression and increased cellular invasion in vitro. Our data indicate that miR-34a could be a potential biomarker for improving the diagnostic efficiency of OC, and miR-34a overexpression may reduce OC pathogenesis by targeting FOXP1.
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2026-09-01 | Brown and white adipocyte like cells derived exosomes differentially regulate stemness, hormone secretion, apoptosis, and lipid metabolism in model cells.
White and brown adipocytes show therapeutic potential, yet their exosomes mediated cell-specific effects remain poorly understood. This study aimed to assess the impact of conditioned media (CM) or exosomes derived from brown and white adipocyte-like cells (BALCs and WALCs, respectively) on four human cell types, including ovarian granulosa cells (OGCs), MCF-7 breast cancer cells, human adipose-derived stem cells (hADSCs), and human umbilical cord mesenchymal stem cells (hUCMSCs). Isolated hADSCs were differentiated into BALCs and WALCs using specific induction media. Corresponding exosomes were isolated from BALCs or WALCs derived CM and characterized. Human cell types-representing stem, somatic, and cancer cells-were treated with CM or exosomes. Cellular responses and lipidomic profiling were evaluated using molecular biology techniques and mass spectrometry, respectively. BALCs-derived exosomes significantly increased estradiol secretion from ovarian granulosa cells (p < 0.05). In MCF-7 cells, CM had a stronger pro-apoptotic effect than exosomes (>2-fold). In hADSCs, both CM and exosomes promoted osteogenic differentiation (2.3-fold) and reduced stemness markers proteins Oct4 and Sox2 (p < 0.05). When treated with BALCs-derived exosomes, hADSCs displayed the highest number of differentially expressed lipids, primarily upregulated hits. Pathway analysis indicated that BALCs-derived exosomes promoted lipid storage and membrane remodeling in hADSCs, enhanced energy metabolism in granulosa cells, reduced fatty acid oxidation in MCF-7 cells, and decreased lipid storage in hUCMSCs. Our results highlight a potential modulatory role of adipocyte-derived exosomes in intercellular communication. The findings of this study may be applicable in regenerative medicine, metabolic regulation and cancer therapeutic approaches.
2026-05-20 | Knockout of filamin A in KGN granulosa tumor cells impairs proliferation, cell cycle progression, migration, and cytoskeletal organization under mechanical stress
Abstract Background Filamin A (FLNA) is an actin-binding protein that regulates mechanosensitivity and functions as an intracellular signaling scaffold in various cell types. It has also been implicated in tumor growth. We recently reported FLNA expression in human ovarian granulosa cells and in KGN cells, a granulosa cell tumor (GCT) line. Results Immunohistochemistry analysis of 51 GCT samples revealed heterogeneous FLNA expression, with approximately 20% showing weak, 18% strong, and the majority moderate expression. We therefore conducted functional studies in KGN cells using CRISPR/Cas9 gene editing. A proteomic approach revealed marked changes in protein abundance upon FLNA depletion: proteins with increased abundance were predominantly related to adhesion, cytoskeletal organization, regulation of cell shape, and lipid metabolic process, whereas those with decreased abundance were associated with DNA replication, cell division, and cell cycle regulation. FLNA-knockout cells showed enlarged cell sizes, reduced proliferation, and slightly affected steroidogenesis. Disruption of FLNA further reduced migration velocity, altered actin cytoskeletal alignment under flow, and modified expression of genes involved in cytoskeletal architecture, adhesion, and mechanosensing under shear stress. Conclusions Our results identify crucial roles of FLNA in shaping the cellular architecture, motility, and proliferation of KGN cells. Consequently, alterations in FLNA expression may influence intracellular signaling, and responsiveness to mechanical cues in both physiological and pathological contexts.
2025-09-18 | Fertility Preservation in an 18-Year-Old Female with an Ovarian Granulosa Cell Tumor: A Case Report
Objective: Ovarian granulosa cell tumors (GCTs) are rare malignancies that can profoundly affect fertility, particularly in young women. This case report describes a successful fertility preservation strategy in an 18-year-old female diagnosed with a Stage I ovarian granulosa cell tumor. Methods: A multidisciplinary team implemented a fertility preservation protocol involving preoperative ovarian stimulation, oocyte retrieval, and cryopreservation. This was followed by staging laparotomy and unilateral salpingo-oophorectomy. Results: A total of 10 mature oocytes were successfully cryopreserved. Histology confirmed a Stage I granulosa cell tumor with no evidence of metastasis. The patient tolerated adjuvant therapy well and remains disease-free at follow-up. Conclusion: The successful application of controlled ovarian stimulation and oocyte cryopreservation prior to definitive surgery demonstrates the feasibility of proactive fertility preservation in hormonally active tumors when managed with careful monitoring. Furthermore, the patient’s positive postoperative course and continued disease-free status at 18 months reinforce the safety and efficacy of fertility-sparing surgical strategies in Stage I GCTs.
2025-05-03 | Mitigating chemotherapy-induced granulosa cell damage: role of hUCMSC-EVs in regulating the lncRNA HCP5-miR-20a-5p-YAP1 network.
The substantial apprehension facing young cancer patients revolves around the onset of chemotherapy-induced premature ovarian failure (POF), primarily linked to damage inflicted upon granulosa cells (GCs). The inquiry delves into the protective role of extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUCMSCs) in mitigating chemotherapy-induced ovarian failure. Specifically, we investigated the mechanism by which hUCMSC-EVs deliver the long non-coding RNA (lncRNA) HCP5 to regulate DNA damage repair in GCs via the miR-20a-5p/YAP1 axis. The detection of differentially expressed lncRNAs in GC injury resulting from cyclophosphamide (CP) was conducted through transcriptome sequencing. hUCMSC-EVs were isolated, characterized, and co-cultured with CP-injured GCs. Functional assays such as CCK-8, TUNEL, and ELISA were performed to evaluate GC viability, apoptosis, and ovarian endocrine function. Experimental validation of the interactions involving HCP5, miR-20a-5p, and YAP1 was achieved through performing luciferase reporter assays, RNA immunoprecipitation experiments, and Western blot (WB) analyses. HCP5 was significantly enriched in hUCMSC-EVs and effectively delivered into GCs. This resulted in improved GC viability, reduced apoptosis, and enhanced DNA repair. Mechanistically, HCP5 sponged miR-20a-5p, leading to the upregulation of YAP1, which in turn mitigated CP-induced GC damage. In vivo experiments further demonstrated that hUCMSC-EVs prevented CP-induced POF through modulation of the HCP5-miR-20a-5p-YAP1 axis. Our research underscores the therapeutic potential of hUCMSC-EVs in delivering HCP5 to promote DNA repair in GCs, thereby preventing chemotherapy-induced POF. This study provides a novel molecular framework for future therapeutic strategies aimed at protecting ovarian function during chemotherapy.
2025-04-03 | Granulosa Cell Death is a Significant Contributor to DNA-Damaging Chemotherapy-Induced Ovarian Insufficiency.
Typically, DNA-damaging chemotherapy (CTx) regimens have a gonadotoxic effect and cause premature ovarian insufficiency (POI), characterized by infertility and estrogen deficiency. However, whether loss of granulosa cells killed directly by CTx contributes significantly to POI has not been determined. To address this issue, we used a previously established mouse model of CTx-induced POI. The alkylating drugs Busulfan (8.75 mg/kg) and Cyclophosphamide (100 mg/kg) were administered to 8-week-old FVB female mice by intraperitoneal (i.p.) injection three times at 48-hour intervals, after which ovarian tissues were harvested and examined by immunofluorescence. The number of primordial follicles was significantly reduced at day (d)6, whereas the number of growing follicles was relatively unchanged. CTx led to DNA double strand breaks in both oocytes and granulosa cells based on the presence of γH2AX foci. However, markers of apoptosis predominantly labeled granulosa cells in growing follicles. We next examined the effect of inhibiting apoptosis in growing granulosa cells by generating Bak-/-Baxfxfx; Cyp19a1Cre transgenic mice. On d10 after the first CTx, Bak-/-Baxfxfx; Cyp19a1Cre ovaries had fewer apoptotic granulosa cells and more surviving follicles than controls. Furthermore, Bak-/-Baxfxfx; Cyp19a1Cre mice showed better fertility than controls after CTx. Our data suggest that granulosa cell death is a significant contributor to follicle depletion and fertility loss after Cyclophosphamide and Busulfan.
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Drug Discovery Landscape
2 orphan drug designations for Malignant granulosa cell tumor of the ovary.
2 orphan drug designations for Malignant granulosa cell tumor of the ovary.
Drug | Therapy type | Orphan designation | Clinical trial status | Sponsor |
|---|---|---|---|---|
Megestrol acetate | small molecules | EMA | — | Grupo Español de Tumores Huérfanos e Infrecuentes (GETHI) |
Ketoconazole | small molecules | EMA | Grupo Español de Tumores Huérfanos e Infrecuentes (GETHI) |
“Terminated” – programs with discontinued clinical trials, or with no updates reported for 5 years after completion or 12 years after the last trial started.
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