We aim to provide a translational model to investigate the reproductive consequences of pubertal delay using the GnRH agonist triptorelin in transgender girls, tested in particular on testicular maturation in peripubertal rats. A total of 30 Sprague Dawley rats were utilized, with 10 subjects assigned to each of three groups from day P30 postpartum (prepubertal) until day P95 (postpubertal), mimicking treatment timing in patients. Rats received triptorelin at three time points (P30, P50, and P71), or only at P30 and P50. Control rats were injected with vehicle. Plasma testosterone levels were determined using MRM analysis. Testes and epididymides were examined histologically. There were significantly lower testosterone levels at postnatal day 48 in treated rats, indicating delayed puberty, with further reductions by day 69. By day 93, testosterone levels had recovered in rats given vehicle at P71 but remained low in the triptorelin-continuous group, suggesting the reversibility of the treatment. Treated rats had smaller testes; however, the majority of the testicular parenchyma was unaffected, with most seminiferous tubules displaying complete spermatogenesis. However, focal atrophic changes were observed in 1–30% of the parenchyma. One-third of the short-term group and half of the long-term group were classified as atrophic. Despite these changes, all treated rats had mature sperm in the epididymis, ensuring their fertility. In conclusion, triptorelin treatment promotes a decline in testosterone levels accompanied by discrete atrophy of the seminiferous tubules, which is partially reversible and compatible with sperm production and fertility preservation. Triptorelin could be an appropriate treatment prior to estrogen therapy for patients seeking gender transition.

The World Health Organization estimates that approximately 285 million people suffer from visual impairments, around 5% of which are caused by corneal pathologies. Currently, the most common clinical treatment consists of a corneal transplant (keratoplasty) from a human donor. However, worldwide demand for donor corneas amply exceeds the available supply. Lamellar keratoplasty (transplantation replacement of only one of the three layers of the cornea) is partially solving the problem of cornea undersupply. Obviously, cell therapy applied to every one of these layers will expand current therapeutic options, reducing the cost of ophthalmological interventions and increasing the effectiveness of surgery. Mesenchymal stem cells (MSCs) are adult stem cells with the capacity for self-renewal and differentiation into different cell lineages. They can be obtained from many human tissues, such as bone marrow, umbilical cord, adipose tissue, dental pulp, skin, and cornea. Their ease of collection and advantages over embryonic stem cells or induced pluripotent stem cells make them a very practical source for experimental and potential clinical applications. In this review, we focus on recent advances using MSCs from different sources to replace the damaged cells of the three corneal layers, at both the preclinical and clinical levels for specific corneal diseases.

When limbal stem cell deficiency (LSCD) is partial, the standard treatment involves covering the corneal surface with amniotic membrane (AM), which supports the proliferation of the remaining limbal stem cells (LSCs). In cases of complete LSCD, the most common treatment is cultured limbal epithelial transplantation (CLET), although there is a risk of rejection. Studies have shown that mesenchymal stem cell transplantation is equally safe and effective as CLET. Recent research has demonstrated successful differentiation ofadipose-derived adult mesenchymal stem cells (ADSCs) into LSCs. Combining AM transplantation with LSCs improves treatment efficacy. However, a limitation of AM use is donor variability and the associated risk of immune rejection. We propose the use of 3D-printed collagen as a scaffold seeded with LSCs derived from ADSCs for the treatment of LSCD in a rat model. The 3D-printed collagen scaffolds exhibited good transparency. In vitro differentiation of ADSCs into LSCs showed morphological changes that were more pronounced and occurred more rapidly on 3D-printed collagen. Among the tested substrates, 3D-printed collagen was the most efficient for differentiation, yielding the highest expression of LSC-specific markers (p63 alpha and BMI-1) and the corneal epithelial marker (SSEA-4). LSCs differentiated in either AM or 3D-printed collagen I scaffolds were transplanted into a rat model of LSCD and compared with the standard, cell-free AM treatment. In all treatment groups, the induced epithelial wound was closed; however, integration of the 3D-printed collagen scaffold was statistically superior to that of AM. However, markers for different corneal structures (PAS, BMI-1, p63 alpha, and cytokeratins 12 and 13) indicated that the generated epithelium was conjunctival rather than corneal, suggesting that the contribution of ADSC-derived LSCs was insufficient for complete corneal re-epithelization.

Corneal opacity remains a leading cause of global blindness, yet conventional corneal transplantation is constrained by donor scarcity, surgical limitations, and suboptimal long-term outcomes. In response, regenerative strategies are advancing to restore structural and functional integrity across all three corneal layers-epithelium, stroma, and endothelium-through cell-based and bioengineered therapies. Among these, induced pluripotent stem cells (iPSCs) have emerged as a versatile and scalable source capable of generating corneal-like cells under defined, xeno-free conditions. This review provides a concise summary of recent clinical progress in corneal cell therapy and tissue engineering, while placing major emphasis on systematically updating preclinical advances in iPSC-based corneal regeneration. Additionally, we highlight the only two first-in-human trials employing iPSC-derived corneal cells, which demonstrate encouraging early safety and efficacy outcomes, yet underscore critical translational challenges including tumorigenicity, immunogenicity, and heterogeneity. Emerging solutions-such as HLA-matched iPSC banks, gene editing, and naïve-state reprogramming-are discussed as key strategies to enhance clinical readiness and scalability.

When limbal stem cell deficiency (LSCD) is partial, the usual treatment is covering the corneal surface with amniotic membrane (AM), allowing the growth of the remaining limbal stem cells (LSCs). In cases of complete LSCD, the most common treatment is cultured limbal epithelial transplantation (CLET), with the possibility of rejection. Studies proved that mesenchymal stem cell (MSC) transplantation is as safe and effective as CLET. Recent studies demonstrate a successful differentiation of adipose-derived adult mesenchymal stem cells (ADSCs) to LSCs. Combining AM transplantation with LSCs results in improved treatment efficacy. A disadvantage of using AM is the variability among donors and the risk of rejection. We propose the use of 3D-printed collagen as a scaffold colonized with LSCs derived from ADSCs for the treatment of LSCD in a rat animal model. The 3D-printed collagen scaffolds showed good transparency. In vitro ADSC differentiation to LSCs showed a change in cell morphology, being more pronounced and faster on 3D-printed collagen. 3D-printed collagen was the most efficient differentiation substrate, with the highest percentage of LSC specific markers (p63α and BMI-1) and corneal epithelium (SSEA-4). The differentiated LSCs in AM or 3D-printed collagen I scaffolds were transplanted into a rat model of LSCD, compared to the standard cell free AM. In all groups the epithelium closed the imposed wound. 3D-printed collagen scaffold integration was statistically superior to AM. Markers for different corneal structures (PAS, BMI-1, p63α, cytokeratins 12 and 13) showed that the generated epithelium was not epithelial but conjunctival, and that the contribution of ADSC-derived LSCs was not enough for re-epithelization of the cornea.

Purpose This study investigates immune cell (ICs) infiltration in advanced keratoconus patients undergoing autologous adipose-derived adult stem cell (ADASC) therapy with recellularized human donor corneal laminas (CL). Methods A prospective clinical trial included fourteen patients divided into three groups: G-1, ADASCs; G-2, decellularized CL (dCL); and G-3, dCL recellularized with ADASCs (ADASCs-rCL). Infiltrated ICs were assessed using in vivo confocal microscopy (IVCM) at 1,3,6, and12 months post-transplant. Results Infiltrated ICs, encompassing granulocytes and agranulocytes, were observed across all groups, categorized by luminosity, structure, and area. Stromal ICs infiltration ranged from 1.19% to 6.62%, with a consistent increase in group-related cell density ( F = 10.68, P < .0001), independent of post-op time ( F = 0.77, P = 0.511); the most substantial variations were observed in G-3 at 6 and 12 months (2.0 and 1.87-fold, respectively). Similarly, significant size increases were more group-dependent ( F = 5.76, P < .005) rather than time-dependent ( F = 2.84, P < .05); G-3 exhibited significant increases at 6 and 12 months (3.70-fold and 2.52-fold, respectively). A lamina-induced shift in IC size occurred ( F = 110.23, P < .0001), primarily with 50–100 μm 2 sizes and up to larger cells > 300μm 2 , presumably macrophages, notably in G-3, indicating a potential role in tissue repair and remodeling, explaining reductions in cells remnants < 50μm 2 . Conclusions ADASCs-rCL therapy may lead to increased IC infiltration compared to ADASCs alone, impacting cell distribution and size due to the presence of the lamina. The findings reveal intricate immune patterns shaped by the corneal microenvironment and highlight the importance of understanding immune responses for the development of future therapeutic strategies.

Cellular therapy of the corneal stroma, either with ocular or extraocular stem cells, has gained significant interest over the last decade. Multiple studies showed benefits such as improving or alleviating corneal scars, improving corneal transparency, generating new collagen within the host stroma, and immunosuppressive and immunomodulatory properties. Mesenchymal stem cells, specifically adipose-derived adult stem cells (ADASCs), differentiate into functional adult keratocytes in vivo and in vitro. Alió et al. have carried out the first clinical, interventional prospective, consecutive, randomized, comparative series of cases on corneal stroma in 14 patients with advanced keratoconus. Patients were divided into three experimental groups. Group-1 (G-1) patients underwent implantation of (ADASCs) alone. Group-2 (G-2) patients received decellularized donor corneal stroma lamina (120 μm thickness). Group-3 (G-3) patients received implantation of donor lamina recellularized with (ADASCs). Autologous ADASCs were obtained by elective liposuction. Under topical anesthesia, implantation was performed into a femtosecond-assisted 9.5-mm diameter lamellar pocket. Follow-up data for 36 months were obtained. This chapter discusses the results of the evolution of corneal stroma cellularity, the morphological evolution of the implanted ADASCs, and the increase in cellularity associated with keratocyte morphological changes, all studied in vivo by using corneal confocal microscopy, together with corneal transparency, and corneal densitometry in patients with implanted ADASCs alone, which we named group A (G-A), and implanted recellularized laminas with ADASCs that have been the subject of a corneal regeneration experience that we named group B (G-B). The promising results obtained in this first human clinical experience of advanced therapy of the cornea with autologous mesenchymal stem cells indicate that we should expect from this new therapy a tremendous development in the following years in the treatment of cornea stromal disease.

Patients with chronic hypoxia show a higher tumor incidence; however, no primary common cause has been recognized. Given the similarities between cellular reprogramming and oncogenic transformation, we directly compared these processes in human cells subjected to hypoxia. Mouse embryonic fibroblasts were employed as controls to compare transfection and reprogramming efficiency; human adipose-derived mesenchymal stem cells were employed as controls in human cells. Easily obtainable human peripheral blood mononuclear cells (PBMCs) were chosen to establish a standard protocol to compare cell reprogramming (into induced pluripotent stem cells (iPSCs)) and oncogenic focus formation efficiency. Cell reprogramming was achieved for all three cell types, generating actual pluripotent cells capable for differentiating into the three germ layers. The efficiencies of the cell reprogramming and oncogenic transformation were similar. Hypoxia slightly increased the reprogramming efficiency in all the cell types but with no statistical significance for PBMCs. Various PBMC types can respond to hypoxia differently; lymphocytes and monocytes were, therefore, reprogrammed separately, finding a significant difference between normoxia and hypoxia in monocytes in vitro. These differences were then searched for in vivo. The iPSCs and oncogenic foci were generated from healthy volunteers and patients with chronic obstructive pulmonary disease (COPD). Although higher iPSC generation efficiency in the patients with COPD was found for lymphocytes, this increase was not statistically significant for oncogenic foci. Remarkably, a higher statistically significant efficiency in COPD monocytes was demonstrated for both processes, suggesting that physiological hypoxia exerts an effect on cell reprogramming and oncogenic transformation in vivo in at least some cell types.

In adults, the limbal stem cells (LSC) reside in the limbal region of the eye, at the junction of the cornea and the sclera where they renew the outer epithelial layer of the cornea assuring transparency. LSC deficiencies (LSCD) due to disease or injury account for one of the major causes of blindness. Among current treatments for LSCD, cornea transparency can be restored by providing new LSC to the damaged eye and induced pluripotent stem cells (iPSC) holds great promise as a new advanced cell source. A synthetic mRNA-based protocol to produce human iPSC from bone marrow mesenchymal stem cells has been defined. The results demonstrate a standardizable method that can be easily adaptable for clinical-grade production standards, produce high-purity LSC-like cells in a relatively rapid timeframe of 12 days, and can be successfully seeded on amniotic membrane or a biodegradable fibrin gel for transplantation. In vivo data demonstrated it is feasible to transplant the iPSC-LSC fibrin patch. In conclusion, an efficient method has been developed to produce patient-specific LSC and seed them on a scaffold fibrin gel for future treatment of LSC-deficiency disease.

Angiosarcoma is a rare soft tissue sarcoma originating from endothelial cells. Given that current treatments for advanced disease have shown limited efficacy, alternative therapies need to be identified. In rare diseases, patient-derived cell models are crucial for screening anti-tumour activity. In this study, cell line models were characterised in 2D and 3D cultures. The cell lines’ growth, migration and invasion capabilities were explored, confirming them as useful tools for preclinical angiosarcoma studies. By screening a drug library, we identified potentially effective compounds: 8-amino adenosine impacted cell growth and inhibited migration and invasion at considerably low concentrations as a single agent. No synergistic effect was detected when combining with paclitaxel, gemcitabine or doxorubicin. These results suggest that this compound could be a potentially useful drug in the treatment of AGS.

Background: Iodine is required for the synthesis of thyroid hormone (TH), but its natural availability is limited. Dehalogenase1 (Dehal1) recycles iodine from mono- and diiodotyrosines (MIT, DIT) to sustain TH synthesis when iodine supplies are scarce, but its role in the dynamics of storage and conservation of iodine is unknown. Methods: Dehal1-knockout (Dehal1KO) mice were generated by gene trapping. The timing of expression and distribution was investigated by X-Gal staining and immunofluorescence using recombinant Dehal1-beta-galactosidase protein produced in fetuses and adult mice. Adult Dehal1KO and wild-type (Wt) animals were fed normal and iodine-deficient diets for 1 month, and plasma, urine, and tissues were isolated for analyses. TH status was monitored, including thyroxine, triiodothyronine, MIT, DIT, and urinary iodine concentration (UIC) using a novel liquid chromatography with tandem mass spectrometry method and the Sandell-Kolthoff (S-K) technique throughout the experimental period. Results: Dehal1 is highly expressed in the thyroid and is also present in the kidneys, liver, and, unexpectedly, the choroid plexus. In vivo transcription of Dehal1 was induced by iodine deficiency only in the thyroid tissue. Under normal iodine intake, Dehal1KO mice were euthyroid, but they showed negative iodine balance due to a continuous loss of iodotyrosines in the urine. Counterintuitively, the UIC of Dehal1KO mice is twofold higher than that of Wt mice, indicating that S-K measures both inorganic and organic iodine. Under iodine restriction, Dehal1KO mice rapidly develop profound hypothyroidism, while Wt mice remain euthyroid, suggesting reduced retention of iodine in the thyroids of Dehal1KO mice. Urinary and plasma iodotyrosines were continually elevated throughout the life cycles of Dehal1KO mice, including the neonatal period, when pups were still euthyroid. Conclusions: Plasma and urine iodotyrosine elevation occurs in Dehal1-deficient mice throughout life. Therefore, measurement of iodotyrosines predicts an eventual iodine shortage and development of hypothyroidism in the preclinical phase. The prompt establishment of hypothyroidism upon the start of iodine restriction suggests that Dehal1KO mice have low iodine reserves in their thyroid glands, pointing to defective capacity for iodine storage.

PURPOSE: This study evaluated 1-year safety and efficacy outcomes of corneal stroma cell therapy. Therapy consisted of implanting autologous adipose-derived adult stem cells (ADASc) with or without sheets of decellularized donor human corneal stroma within the stroma of patients with advanced keratoconus. DESIGN: This was a prospective interventional non-randomized series of cases. METHODS: Fourteen consecutive patients were selected and divided into 3 experimental groups. Group A patients underwent implantation of autologous ADASc alone (3 x 10(6) cells/1 mL) (n = 5). Group B patients received decellularized donor 120-mu m-thick corneal stroma lamina alone (n = 5). Group C patients had implantation of recellularized donor lamina with 1 x 10(6) autologous ADASc plus another 1 x 10(6) cells/1 mL at the time of the surgery (n = 4). Autologous ADASc were obtained by elective liposuction. Implantation was performed in the corneal stroma through a femtosecond-assisted 9.5-mm diameter lamellar dissection with the patient under topical anesthesia. Twelve months of follow-up data are presented. RESULTS: No complications were observed during the 1-year follow-up, and full corneal transparency was recovered within 3 months in all patients. No patient lost lines of visual acuity. Corrected distance visual acuity improved 0.231, 0.264, and 0.094 Snellen lines in groups 1, 2, and 3, respectively. In group 1, refractive parameters showed an overall stability, whereas in groups 2 and 3, sphere improved 2.35 diopter (D) and 0.625 D, respectively. Anterior keratometry remained stable (group 1) and improved in groups 2 and 3 (mean improvement of 2D). Corneal aberrometry improved significantly. In optical coherence tomography scans, corneal thickness showed a mean improvement of 14.5 pi.m (group 1) and 116.4 mu m (groups 2 and 3) in the central thickness, and new collagen production was observed at the surgical plane (group 1). Confocal biomicroscopy confirmed the host recellularization of the implanted laminas. CONCLUSIONS: Intrastromal implantation of autologous ADASc and decellularized human corneal stroma did not show complications at 1 year of follow-up and were moderately effective for the treatment of advanced keratoconus. NOTE: Publication of this article is sponsored by the American Ophthalmological Society. (C) 2019 Elsevier Inc. All rights reserved.

Approximately 20–30% of endometrial carcinomas (EC) are characterized by mismatch repair (MMR) deficiency (dMMR) or microsatellite instability (MSI), and their testing has become part of the routine diagnosis. The aim of this study was to establish and compare the MMR status using various approaches. Immunohistochemistry (IHC), PCR-based MSI, and the detection of defects in the four key MMR genes (MLH1, PMS2, MSH2, and MSH6) via methylation-specific multiplex ligation-dependent probe amplification (MLPA) and targeted next-generation sequencing (NGS) were performed. MSH3 expression was also evaluated. A set of 126 early-stage EC samples were analyzed, 53.2% of which were dMMR and 46.8% of which were proficient MMR (pMMR) as determined using IHC, whereas 69.3% were classified as microsatellite stable, while 8.8% and 21.9% were classified MSI-low (MSI-L) and MSI-high (MSI-H), respectively. In total, 44.3% of the samples showed genetic or epigenetic alterations in one or more genes; MLH1 promoter methylation was the most common event. Although acceptable concordance was observed, there were overall discrepancies between the three testing approaches, mainly associated with the dMMR group. IHC had a better correlation with MMR genomic status than the MSI status determined using PCR. Further studies are needed to establish solid conclusions regarding the best MMR assessment technique for EC.

Approximately 10 million individuals have blindness due to limbal stem cell (LSCs) deficiency, one of the most challenging problems in ophthalmology. To replenish the LSC pool, an autologous extraocular cell source is appropriate, thereby avoiding the risk of immune rejection, the need for immunosuppression and the risk of damaging the contralateral eye. In recent years, adipose-derived mesenchymal stem cells (ADSCs) have been a key element in ocular regenerative medicine. In this study, we developed a protocol for deriving human LSCs from ADSCs compatible with the standard carrier human amniotic membrane, helping provide a stem cell pool capable of maintaining proper corneal epithelial homeostasis. The best protocol included an ectodermal induction step by culturing ADSCs with media containing fetal bovine serum, transforming growth factor-β inhibitor SB-505124, Wnt inhibitor IWP-2 and FGF2 for 7 days, followed by an LSC induction step of culture in modified supplemental hormonal epithelial medium supplemented with pigment epithelium-derived factor and keratinocyte growth factor for 10 additional days. The optimal differentiation efficiency was achieved when cells were cultured in this manner over vitronectin coating, resulting in up to 50% double-positive αp63/BMI-1 cells. The results of this project will benefit patients with LSC deficiency, aiding the restoration of vision.

The human cornea is a five-layered tissue that provides two-thirds of the total refractive power of the eye, and it is the first barrier protecting the intraocular content. The corneal endothelium, the inner layer, is in charge of maintaining the cornea in a relatively dehydrated state and therefore transparent. The endothelial cell layer failure leads to corneal swelling, loss of transparency and blindness. Currently, the only effective and probed way to restore endothelial function universally is to perform an allogenic graft. Since Melles revolutionized the field by describing a method to dissect only Descemet Membrane (DM) from the recipient eye, leaving the posterior lamella intact, and after Price and Gorovoy pioneered Descemet stripping endothelial keratoplasty (DSEK), a variety of endothelial keratoplasty techniques have taken over. However, there is a scarcity of donors to adequate to high and increasing demand. Cell culture techniques make it possible to expand ex vivo the corneal endothelial cells (CEC) to subsequently inject a cell solution into the anterior chamber, or else to manufacture constructs made up of acellular corneal stroma, acellular Descemet membrane or carriers manufactured by tissue bioengineering, and colonized by CEC expanded ex vivo, which could then be grafted onto the recipient. Nowadays, we are in an outstanding position to develop corneal endothelial cell sheets for endothelial keratoplasty: reproducible and well-defined culturing methods and conditions have been achieved in the last decades. Regardless of advances in promoting human CEC proliferation, the achieved capacity for expanding human CECs is still highly limited; new sources of CECs are therefore sought. The use of extraocular cells capable of differentiating into corneal endothelial cells is highly desirable. Recent advances have been achieved in differentiation protocols from embryonic stem cells and adipose-derived mesenchymal stem cells. New advances in biomimetic materials and manufacturing protocols such as electrospinning, nanolithography, vitrification, and advances in novel 3D printing techniques such as LIFT, laser-assisted bioprinting, and others will aid in the search for a donor-independent biocompatible carrier. Further development of these and previous approaches, by defining the growth factors, the signaling pathways implicated in directed differentiation, the use of more practical cells to derive hCECs, and the in vivo demonstration of functionality are urgently needed.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with an overall 5-year survival rate of just 5%. A better understanding of the carcinogenesis processes and the mechanisms of the progression of PDAC is mandatory. Fifty-two PDAC patients treated with surgery and adjuvant therapy, with available primary tumors, normal tissue, preneoplastic lesions (PanIN), and/or lymph node metastases, were selected for the study. Proteins were extracted from small punches and analyzed by LC-MS/MS using data-independent acquisition. Proteomics data were analyzed using probabilistic graphical models, allowing functional characterization. Comparisons between groups were made using linear mixed models. Three proteomic tumor subtypes were defined. T1 (32% of patients) was related to adhesion, T2 (34%) had metabolic features, and T3 (34%) presented high splicing and nucleoplasm activity. These proteomics subtypes were validated in the PDAC TCGA cohort. Relevant biological processes related to carcinogenesis and tumor progression were studied in each subtype. Carcinogenesis in the T1 subtype seems to be related to an increase of adhesion and complement activation node activity, whereas tumor progression seems to be related to nucleoplasm and translation nodes. Regarding the T2 subtype, it seems that metabolism and, especially, mitochondria act as the motor of cancer development. T3 analyses point out that nucleoplasm, mitochondria and metabolism, and extracellular matrix nodes could be involved in T3 tumor carcinogenesis. The identified processes were different among proteomics subtypes, suggesting that the molecular motor of the disease is different in each subtype. These differences can have implications for the development of future tailored therapeutic approaches for each PDAC proteomics subtype.

Adipose-derived stem cells are a subtype of mesenchymal stem cell that offers the important advantage of being easily obtained (in an autologous manner) from low invasive procedures, rendering a high number of multipotent stem cells with the potential to differentiate into several cellular lineages, to show immunomodulatory properties, and to promote tissue regeneration by a paracrine action through the secretion of extracellular vesicles containing trophic factors. This secretome is currently being investigated as a potential source for a cell-free based regenerative therapy for human tissues, which would significantly reduce the involved costs, risks and law regulations, allowing for a broader application in real clinical practice. In the current article, we will review the existing preclinical and human clinical evidence regarding the use of such adipose-derived mesenchymal stem cells for the regeneration of the three main layers of the human cornea: the epithelium (derived from the surface ectoderm), the stroma (derived from the neural crest mesenchyme), and the endothelium (derived from the neural crest cells).

There are three prognostic stratification tools used for endometrial cancer: ESMO-ESGO-ESTRO 2016, ProMisE, and ESGO-ESTRO-ESP 2020. However, these methods are not sufficiently accurate to address prognosis. The aim of this study was to investigate whether the integration of molecular classification and other biomarkers could be used to improve the prognosis stratification in early-stage endometrial cancer. Relapse-free and overall survival of each classifier were analyzed, and the c-index was employed to assess accuracy. Other biomarkers were explored to improve the precision of risk classifiers. We analyzed 293 patients. A comparison between the three classifiers showed an improved accuracy in ESGO-ESTRO-ESP 2020 when RFS was evaluated (c-index = 0.78), although we did not find broad differences between intermediate prognostic groups. Prognosis of these patients was better stratified with the incorporation of CTNNB1 status to the 2020 classifier (c-index 0.81), with statistically significant and clinically relevant differences in 5-year RFS: 93.9% for low risk, 79.1% for intermediate merged group/CTNNB1 wild type, and 42.7% for high risk (including patients with CTNNB1 mutation). The incorporation of molecular classification in risk stratification resulted in better discriminatory capability, which could be improved even further with the addition of CTNNB1 mutational evaluation.

Ovarian cancer (OC) is a life-threatening tumor and the deadliest among gynecological cancers in developed countries. First line treatment with a carboplatin/paclitaxel regime is initially effective in the majority of patients, but most advanced OC will recur and develop drug resistance. Therefore, the identification of alternative therapies is needed. In this study, we employed a panel of high-grade serous ovarian cancer (HGSOC) cell lines, in monolayer and three-dimensional cell cultures. We evaluated the effects of a novel tubulin-binding agent, plocabulin, on proliferation, cell cycle, migration and invasion. We have also tested combinations of plocabulin with several drugs currently used in OC in clinical practice. Our results show a potent antitumor activity of plocabulin, inhibiting proliferation, disrupting microtubule network, and decreasing their migration and invasion capabilities. We did not observe any synergistic combination of plocabulin with cisplatin, doxorubicin, gemcitabine or trabectedin. In conclusion, plocabulin has a potent antitumoral effect in HGSOC cell lines that warrants further clinical investigation.