Deciphering brain metastasis in epithelial ovarian cancer: multimodal analysis and potential biomarkers R. Trozzi, M. Salvi, M. Karimi, A. Minucci, G. Raspaglio, M. De Donato, M. Buttarelli, A. Piermattei, L. Vaccaro, A. Grimaldi, R. De Santis, M. Massa, F. Sillano, L. Giacò, L. Mastrantoni, V. Iacobelli, F. Camarda, M. Cesana, S. Duranti, M. C. Sassu, P. Mattogno, A. Fagotti, C. Marchetti, G. Scambia, C. Nero, D. Cacchiarelli Npj Precision Oncology, 2026 Epithelial ovarian cancer (EOC) remains the most lethal gynaecological malignancy in developed countries, with recurrence and drug resistance posing significant clinical challenges. Brain metastases (BM) from epithelial ovarian cancer, once rare, are an increasing phenomenon and are characterised by a dismal prognosis. To explore the molecular underpinnings of BM in EOC, we conducted a multimodal genomics and transcriptomics analysis of matched primary tumour and brain metastases samples from a retrospective cohort. Our findings revealed high genomic concordance between primary tumour (PT) and BM, with alterations in key pathways such as MYC (MYC Proto-Oncogene, bHLH Transcription Factor) targets, extracellular matrix remodelling, and inflammatory signalling characterizing the BM. AFP (Alpha-fetoprotein) and GFAP (Glial Fibrillary Acidic Protein) emerged as potential biomarkers from the primary lesion for BM onset, while network analysis identified MET (MET Proto-Oncogene, Receptor Tyrosine Kinase), GDF15 (Growth Differentiation Factor 15), and S100A9 (S100 Calcium Binding Protein A9) as candidate mediators of tumour-brain crosstalk. These results offer new insights into EOC brain tropism, highlighting potential targets for therapeutic intervention and personalized patient management in the precision oncology era.
Co-targeting hallmarks of cancer for therapeutic benefit in ovarian cancer: a scoping review Valentina Iacobelli, Floriana Camarda, Gloria Anderson, Marianna Buttarelli, Luca Mastrantoni, Miriam Grazia Ferrara, Rita Trozzi, Simona Duranti, Vanda Salutari, Giulia Sabetta, Giovanni Scambia, Anna Fagotti, Camilla Nero International Journal of Gynecological Cancer, 2026 The conceptualization of cancer characteristics into 14 hallmarks is now widely adopted. Simultaneous targeting of multiple cancer hallmarks represents a promising strategy to overcome therapeutic resistance and improve clinical outcomes. This approach is particularly relevant in epithelial ovarian cancer, which remains highly lethal despite significant advances in first-line treatment. This scoping review was conducted according to Preferred Reporting Items for Systematic reviews and Meta-Analyses Extension for Scoping Reviews (PRISMA-ScR) guidelines and included searches of Medline, Scopus, the Cochrane Library, and ClinicalTrials.gov for studies published up to September 30, 2024. To be eligible, trials were required to be phase I to III clinical trials (both completed and ongoing) enrolling patients with any histotype of epithelial ovarian cancer and to target ≥2 of the 14 cancer hallmarks. Studies were screened and data extracted independently by 3 reviewers. Out of 1461 records screened at the title and abstract level (data cutoff: September 30, 2024), 225 studies were identified, comprising 111 completed and 114 ongoing trials. The adoption of co-targeting strategies has notably increased, with a 3-fold increase in trials between 2007-2013 and 2021-2024. Among 94 trials reporting clinically meaningful benefits, the most frequent combination involved targeting "sustaining proliferative signaling" and "genome instability and mutations." In ongoing trials, 40 are focused on modulating "avoiding immune destruction." However, no clinical trials were identified for 4 of the 14 hallmarks: "unlocking phenotypic plasticity," "senescent cells," "non-mutational epigenetic re-programming," and "polymorphic microbiomes." These hallmarks remain underexplored, highlighting critical gaps and potential areas for future research. In conclusion, co-targeting approaches in epithelial ovarian cancer rely on combining genomic instability and angiogenesis inhibition with chemotherapy. Current research trends are shifting toward chemotherapy-free regimens and novel therapeutic targets, aiming to address resistance mechanisms and improve long-term outcomes.
Description of a Large Family with Periodic Fever Carrying a Variant in RXFP1 Gene: A Possible Novel Modulator of Inflammation in Autoinflammatory Diseases Marianna Buttarelli, Giulia Rapari, Melania Riccio, Raffaele Manna, Donato Rigante, Eugenio Sangiorgi International Journal of Molecular Sciences, 2026 Autoinflammatory diseases involve recurrent systemic inflammation caused by dysregulated innate immunity, arising from genetic or multifactorial mechanisms, as seen in periodic fever, aphthous stomatitis, pharyngitis, and adenitis (PFAPA) syndrome. About 10% of PFAPA patients show autosomal dominant inheritance. We describe a three-generation family with a PFAPA-like recurrent fever syndrome displaying clear autosomal dominant transmission. All affected individuals tested negative on a diagnostic panel of 13 known autoinflammatory genes. Whole-exome sequencing was performed in two distantly related affected members, followed by variant filtering, segregation analysis, and phenotype-based prioritization. A single heterozygous missense variant in RXFP1, c.154G>A p.(Asp52Asn), co-segregated with disease in all affected relatives. This variant is extremely rare in population databases, absent from ClinVar, present in COSMIC, and predicted as damaging by REVEL and CADD. RXFP1, not previously implicated in autoinflammatory or innate immune disorders, encodes the relaxin family peptide receptor 1, a G protein–coupled receptor involved in extracellular matrix regulation, anti-fibrotic pathways, and modulation of inflammatory cytokine production. Protein network analysis showed interactions with RLXN1-3, inflammatory mediators, PTGDR, ADORA2B, and C1QTNF8, supporting an immunomodulatory function. This is the first report linking RXFP1 variation to a hereditary recurrent fever syndrome, identifying relaxin signalling as a potential immune regulatory pathway.
A comparative analysis of tumor markers reveals EDA fibronectin as a promising target in high-grade serous ovarian cancer Alessia Piermattei, Roberto De Luca, Frederik Peissert, Louis Plüss, Emanuele Puca, Nicoletta D’Alessandris, Antonio Travaglino, Francesca Sillano, Tina Pasciuto, Diana Giannarelli, Gian Franco Zannoni, Anna Fagotti, Dario Neri, Giovanni Scambia, Marianna Buttarelli, Camilla Nero Journal of Ovarian Research, 2025 BACKGROUND: Ovarian cancer remains a major clinical challenge with more than 40.000 annual deaths in Europe and in the United States, highlighting the need for better diagnostic and therapeutic strategies. This study first presents an immunohistochemical evaluation of the extra-domains A and B containing fibronectin (EDA-FN, EDB-FN), fibroblast activation protein (FAP), and carcinoembryonic antigen (CEA) in ovarian cancer specimens. Based on the initial results, the analysis was subsequently expanded to provide a comprehensive assessment of EDA-FN expression in human epithelial ovarian cancer tissue samples. METHODS: An initial exploratory immunohistochemical analysis was performed on 60 formalin fixed paraffin embedded (FFPE) epithelial ovarian cancer (EOC) tissue sections from 47 patients, including 47 specimens collected at first diagnosis and 13 matched relapsed lesions. Tissue sections were stained using previously validated antibodies specific to EDA-FN, EDB-FN, FAP and CEA, to evaluate the stromal immunoreactive score (sIRS Part 1). Following the completion of Part 1, the study was expanded to specifically analyze the most abundant antigen found (EDA-FN) on 204 FFPE High Grade Serous ovarian cancer (HGSOC) tissue samples from 102 subjects, including primary and metastatic sites from the same patient (Part 2). RESULTS: In Part 1, stromal expression of EDA-FN, EDB-FN and FAP was observed in epithelial ovarian cancer with no significant differences between matched primary and relapse tumor tissues. CEA was exclusively found in mucinous ovarian cancer (MOC). EDA-FN was the most abundant antigen among the ones investigated, prompting a deeper investigation in Part 2. In the expanded EOC cohort, EDA-FN remained highly abundant across all molecular subgroups (HRp, HRd/BRCAwt, and BRCAmut) and clinical subgroups (naïve vs. pretreated patients), but was found at elevated level in metastases compared to the corresponding primary tumors. CONCLUSIONS: These findings highlight that EDA-FN is an excellent target for HGSOC, while CEA could serve as a potential target for MOC. Clinical investigations are warranted to evaluate innovative treatments in ovarian cancer targeting these antigens.
Design and synthesis of pyridopyrimidines targeting NEK6 kinase Paolo Zardi, Benedetta Righino, Davide Pirolli, Matteo Gramanzini, Alessandro Semeraro, Juan José Galano-Frutos, Anna Königs, Luka Ðorđević, Michele Maggini, Marianna Buttarelli, Natalia Cappoli, Viviana Romano, Marta De Donato, Daniela Gallo, Giovanni Scambia, Maria Cristina De Rosa Archives of Biochemistry and Biophysics, 2025 We designed a series of pyrido[2,3- d ]pyrimidine derivatives based on the structure of the NEK6 kinase inhibitor, compound 21 (2-amino-5-phenyl-5,11-dihydro-3H-indeno[2′,1':5,6]pyrido[2,3- d ]pyrimidine-4,6-dione), which share the same heterocyclic core. Chemical modifications, aimed at altering the molecular planarity of 21 to enhance water solubility, were guided by receptor-based ligand design and further supported by molecular docking, molecular dynamics simulations, and free energy perturbation calculations. Our results indicate that disrupting the planarity of 21 increases aqueous solubility ― nearly doubling it in two cases― while reducing lipophilicity. Among the compounds tested, three showed both improved solubility and NEK6 inhibitory activity exceeding 50 % in single-dose assay. • Pyrido[2,3- d ]pyrimidine derivatives show promising profiles as NEK6 inhibitors. • The ring-opening strategy boosts solubility while retaining NEK6 activity. • FEP simulations and QSAR highlight key features to enhance NEK6 inhibitors' properties.
Decoding tumor evolution in advanced ovarian cancer: Proteogenomic insights before and after neoadjuvant chemotherapy. Valentina Iacobelli, Marianna Buttarelli, Enrica Martinelli, Alessia Piermattei, Giuseppina Raspaglio, Marta De Donato, Francesca Sillano, Rita Trozzi, Floriana Camarda, Tina Pasciuto, Angelo Minucci, Luca Mastrantoni, Luciano Giaco', Giulia Mantini, Eduardo Maria Sommella, Vicky Caponigro, Pietro Campiglia, Gloria Anderson, Giovanni Scambia, Camilla Nero Journal of Clinical Oncology, 2025 e17582 Background: Platinum-based neoadjuvant chemotherapy (NACT) is a cornerstone in advanced ovarian cancer (OC) treatment. However, biomarkers predictive of response and the mechanisms underlying chemoresistance remain poorly defined. The PROGENITOR study aimed to (1) identify biomarkers associated with treatment response, as measured by Chemotherapy Response Score (CRS) and platinum-free interval (PFI), and (2) elucidate pathways driving chemoresistance and tumor adaptation. Methods: Tumor samples from 39 OC patients were analyzed pre- (T0) and post-NACT (T1) using RNA sequencing and liquid chromatography-mass spectrometry. Differentially expressed genes (DEGs) and proteins (DEPs) were identified, and transcriptomic-proteomic integration was performed. Deconvolution analysis, using xCell tool, evaluated tumor microenvironment dynamics, stratified by CRS and PFI. Genomic alterations were also correlated with transcriptomic and proteomic changes. Results: Longitudinal analysis revealed significant downregulation of PKMYT1, CDK1, and UBE2C at T1, suggesting impaired cell cycle progression and the potential induction of a quiescent state, contributing to chemoresistance mechanisms, as observed in CCNE1-amplified tumors. Deconvolution analysis revealed notable shifts in the tumor microenvironment post-NACT. Cancer-associated fibroblasts (CAFs) and stromal scores increased significantly in OC patients with suboptimal response (CRS 1–2), highlighting their role in chemoresistance through extracellular matrix remodeling. Additionally, immune cell populations showed significant alterations, including reduced T-cell and natural killer cell composition, along with a decline in dendritic cells, suggesting impaired antitumor immunity. These immune-stromal alterations were more pronounced in patients with poor PFI (<6 months). Conclusions: This study suggests dependency on PKMYT1 for CDK1 inhibition and a role in chemoresistance, particularly in CCNE1-amplified tumors. The integration of multi-omics and deconvolution analysis underscores the critical roles of cell cycle dysregulation, stromal activation, and immune suppression in shaping tumor evolution under NACT. These findings provide a foundation for biomarker-driven therapeutic strategies targeting cell-cycle machinery.
Correction to: The interaction of β-arrestin1 with talin1 driven by endothelin A receptor as a feature of α5β1 integrin activation in high-grade serous ovarian cancer (Cell Death & Disease, (2023), 14, 1, (73), 10.1038/s41419-023-05612-7) Ilenia Masi, Flavia Ottavi, Danila Del Rio, Valentina Caprara, Cristina Vastarelli, Sara Maria Giannitelli, Giulia Fianco, Pamela Mozetic, Marianna Buttarelli, Gabriella Ferrandina, Giovanni Scambia, Daniela Gallo, Alberto Rainer, Anna Bagnato, Francesca Spadaro, Laura Rosanò Cell Death and Disease, 2024 The authors regret that there is a mistake in Fig. 8B as published in the original article. In the first published version of this manuscript, the bioluminescent image of intraperitoneally (i.p) SKOV3-Luc-injected mice for AMB group was accidentally misused during the assembly of the figures. We greatly apologize for this error and are now providing a corrected version of the figure (see new Fig. 8B). The scientific conclusions of our study are not affected by this inadvertent error.
Dual inhibition of CDK12 and CDK13 uncovers actionable vulnerabilities in patient-derived ovarian cancer organoids Eleonora Cesari, Alessandra Ciucci, Marco Pieraccioli, Cinzia Caggiano, Camilla Nero, Davide Bonvissuto, Francesca Sillano, Marianna Buttarelli, Alessia Piermattei, Matteo Loverro, Floriana Camarda, Viviana Greco, Maria De Bonis, Angelo Minucci, Daniela Gallo, Andrea Urbani, Giuseppe Vizzielli, Giovanni Scambia, Claudio Sette Journal of Experimental and Clinical Cancer Research, 2023 Background High grade serous ovarian cancer (HGSOC) is highly lethal, partly due to chemotherapy resistance and limited availability of targeted approaches. Cyclin dependent kinases 12 and 13 (CDK12/13) are promising therapeutic targets in human cancers, including HGSOC. Nevertheless, the effects of their inhibition in HGSOC and the potential synergy with other drugs are poorly known. Methods We analyzed the effects of the CDK12/13 inhibitor THZ531 in HGSOC cells and patient-derived organoids (PDOs). RNA sequencing and quantitative PCR analyses were performed to identify the genome-wide effects of short-term CDK12/13 inhibition on the transcriptome of HGSOC cells. Viability assays with HGSOC cells and PDOs were performed to assess the efficacy of THZ531 as single agent or in combination with clinically relevant drugs. Results The CDK12 and CDK13 genes are deregulated in HGSOC and their concomitant up-regulation with the oncogene MYC predicts poor prognosis. HGSOC cells and PDOs display high sensitivity to CDK12/13 inhibition, which synergizes with drugs in clinical use for HGSOC. Transcriptome analyses revealed cancer-relevant genes whose expression is repressed by dual CDK12/13 inhibition through impaired splicing. Combined treatment with THZ531 and inhibitors of pathways regulated by these cancer relevant genes (EGFR, RPTOR, ATRIP) exerted synergic effects on HGSOC PDO viability. Conclusions CDK12 and CDK13 represent valuable therapeutic targets for HGSOC. We uncovered a wide spectrum of CDK12/13 targets as potential therapeutic vulnerabilities for HGSOC. Moreover, our study indicates that CDK12/13 inhibition enhances the efficacy of approved drugs that are already in use for HGSOC or other human cancers.
Exploring the Control of PARP1 Levels in High-Grade Serous Ovarian Cancer Giuseppina Raspaglio, Marianna Buttarelli, Natalia Cappoli, Alessandra Ciucci, Anna Fagotti, Giovanni Scambia, Daniela Gallo Cancers, 2023 High-grade serous ovarian cancer (HGSOC) is a leading cause of mortality from gynecologic malignancies worldwide. Although a transformative improvement has been shown with the introduction of PARP (poly(ADP-ribose) polymerase) inhibitors, the emergence of resistance to these drugs represents a therapeutic challenge. Hence, expanding our understanding of mechanisms behind the control of PARP1 expression can provide strategic guidance for the translation of novel therapeutic strategies. The Signal Transducer and Activator of Transcription (STAT) family of proteins consists of transcription factors critically involved in the regulation of important cellular functions. Notably, we recently demonstrated that, in cervical cancer cells, STAT1 controls PARP1 levels through multiple mechanisms, possibly involving also STAT3. Here, we tested the hypothesis that a similar mechanism might be operative in HGSOC. To this end, the impact of STAT1/STAT3 modulation on PARP1 expression was assessed in established and primary HGSOC cells, and molecular biology studies proved that STAT1 might act at both transcriptional and post-transcriptional levels to modulate the PARP1 level. Notably, bioinformatics analysis of TCGA databases demonstrated that increased STAT1 mRNA expression levels are associated with a favorable prognosis and with response to chemotherapy in HGSOC patients. Our findings suggest an alternative strategy for targeting HGSOC cells based on their dependency on PARP1.
The interaction of β-arrestin1 with talin1 driven by endothelin A receptor as a feature of α5β1 integrin activation in high-grade serous ovarian cancer Ilenia Masi, Flavia Ottavi, Danila Del Rio, Valentina Caprara, Cristina Vastarelli, Sara Maria Giannitelli, Giulia Fianco, Pamela Mozetic, Marianna Buttarelli, Gabriella Ferrandina, Giovanni Scambia, Daniela Gallo, Alberto Rainer, Anna Bagnato, Francesca Spadaro, Laura Rosanò Cell Death and Disease, 2023 Dissemination of high-grade serous ovarian cancer (HG-SOC) in the omentum and intercalation into a mesothelial cell (MC) monolayer depends on functional α5β1 integrin (Intα5β1) activity. Although the binding of Intα5β1 to fibronectin drives these processes, other molecular mechanisms linked to integrin inside-out signaling might support metastatic dissemination. Here, we report a novel interactive signaling that contributes to Intα5β1 activation and accelerates tumor cells toward invasive disease, involving the protein β-arrestin1 (β-arr1) and the activation of the endothelin A receptor (ETAR) by endothelin-1 (ET-1). As demonstrated in primary HG-SOC cells and SOC cell lines, ET-1 increased Intβ1 and downstream FAK/paxillin activation. Mechanistically, β-arr1 directly interacts with talin1 and Intβ1, promoting talin1 phosphorylation and its recruitment to Intβ1, thus fueling integrin inside-out activation. In 3D spheroids and organotypic models mimicking the omentum, ETAR/β-arr1-driven Intα5β1 signaling promotes the survival of cell clusters, with mesothelium-intercalation capacity and invasive behavior. The treatment with the antagonist of ETAR, Ambrisentan (AMB), and of Intα5β1, ATN161, inhibits ET-1-driven Intα5β1 activity in vitro, and tumor cell adhesion and spreading to intraperitoneal organs and Intβ1 activity in vivo. As a prognostic factor, high EDNRA/ITGB1 expression correlates with poor HG-SOC clinical outcomes. These findings highlight a new role of ETAR/β-arr1 operating an inside-out integrin activation to modulate the metastatic process and suggest that in the new integrin-targeting programs might be considered that ETAR/β-arr1 regulates Intα5β1 functional pathway.
Clinical value of LncRNA MEG3 in high-grade serous ovarian cancer Marianna Buttarelli, Marta De Donato, Giuseppina Raspaglio, Gabriele Babini, Alessandra Ciucci, Enrica Martinelli, Pina Baccaro, Tina Pasciuto, Anna Fagotti, Giovanni Scambia, Daniela Gallo Cancers, 2020
Estrogens counteract platinum-chemosensitivity by modifying the subcellular localization of MDM4 Rossella Lucà, Giorgia di Blasio, Daniela Gallo, Valentina Monteleone, Isabella Manni, Laura Fici, Marianna Buttarelli, Germana Ciolli, Marsha Pellegrino, Emanuela Teveroni, Silvia Maiullari, Alessandra Ciucci, Alessandro Apollo, Francesca Mancini, Maria Pia Gentileschi, Gian Franco Zannoni, Alfredo Pontecorvi, Giovanni Scambia, Fabiola Moretti Cancers, 2019
A combined ANXA2-NDRG1-STAT1 gene signature predicts response to chemoradiotherapy in cervical cancer Marianna Buttarelli, Gabriele Babini, Giuseppina Raspaglio, Flavia Filippetti, Alessandra Battaglia, Alessandra Ciucci, Gabriella Ferrandina, Marco Petrillo, Carmela Marino, Mariateresa Mancuso, Anna Saran, Maria Elena Villani, Angiola Desiderio, Chiara D’Ambrosio, Andrea Scaloni, Giovanni Scambia, Daniela Gallo Journal of Experimental and Clinical Cancer Research, 2019
