Colorimetric aptasensor for exosome detection in breast cancer liquid biopsy Bartolomeo Della Ventura, Cristina Quintavalle, Erica Cavaliere, Kristin Tkalčec, Paolo Aniello, Vincenzo Iannotti, Gerolama Condorelli, Raffaele Velotta Sensors and Actuators B Chemical, 2026 Exosomes are nanoscale extracellular vesicles (EVs) that carry molecular signatures reflective of their cells of origin, making them attractive biomarkers for liquid biopsy applications. In this study, we present a rapid and highly specific colorimetric aptasensor for the detection of Gremlin-1 (GREM1)-expressing exosomes in serum. The sensing strategy relies on the disaggregation of gold nanoparticle (AuNP) clusters, initially formed by NaCl-induced aggregation, upon selective binding of the target. AuNPs were functionalized with a thiolated Ex.50.T aptamer specifically recognizing exosomal GREM1, and the binding event triggers a measurable spectral shift in the plasmonic profile. A detection limit below 10 6 exosomes/mL was achieved using serial dilutions of purified exosomes isolated from breast cancer serum samples, demonstrating the method’s sensitivity and robustness under controlled conditions. Applied to 100 clinical serum samples, the assay demonstrated excellent diagnostic performance, with 84% sensitivity and 90% specificity—values comparable to those of mammography—without requiring extensive sample processing. Comparative analysis with commercial ELISA and Ex.50.T aptamer-based ELONA confirmed the superior discriminatory power of the method proposed here. Transmission electron microscopy further corroborated the mechanism by revealing exosomes physically disrupting AuNP aggregates. These results highlight the diagnostic potential of exosome-focused sensing strategies and establish this aptamer-based colorimetric platform as a promising candidate for non-invasive screening of breast cancer through liquid biopsy. • Colorimetric aptasensor discriminates breast cancer serum with 84% sensitivity, 90% specificity • Simple optical test matches mammography performance in liquid biopsy for breast cancer • Mix-and-measure detection of tumor-derived exosomes in 1:1 diluted serum • Anti-aggregation assay detects GREM-1-positive exosomes with high diagnostic accuracy • TEM imaging is consistent with exosome-induced disaggregation of gold nanoparticle clusters
Proteomic profiling of extracellular vesicles in ST-elevation acute myocardial infarction Cristina Quintavalle, Giuseppina Roscigno, Gianluca Petrillo, Luca Paolucci, Mario Scarpelli, Amelia Focaccio, Mariateresa Librera, Katia Pane, Monica Franzese, Silvia Nuzzo, Alessandra Affinito, Giuseppe Biondi Zoccai, Francesca De Micco, Zoran Minic, Maxim V Berezovski, Carlo Briguori, Gerolama Condorelli Cardiovascular Research, 2026
Fibroblasts activated by miRs-185-5p, miR-652-5p, and miR-1246 shape the tumor microenvironment in triple-negative breast cancer via PATZ1 downregulation Giada De Luca, Gianluca Petrillo, Iolanda Scognamiglio, Katia Pane, Lorenza Cocca, Giuseppina Roscigno, Martina Mascolo, Claudia Pignataro, Sara Verde, Aurelia Fraticelli, Danilo Fiore, Alessandra Affinito, Silvia Nuzzo, Zoran Minic, Francesca De Micco, Guglielmo Thomas, Monica Franzese, Maxim V. Berezovski, Monica Fedele, Gerolama Condorelli, Cristina Quintavalle Cellular and Molecular Life Sciences, 2025 The intricate interplay between epithelial and fibroblast cells within the tumor microenvironment plays a crucial role in driving triple-negative breast cancer progression. This crosstalk involves the exchange of various signaling molecules, including growth factors, cytokines, extracellular matrix components, and extracellular vesicles. Recently, we demonstrated that triple-negative breast cancer extracellular vesicles carry and release a specific combination of miRs, including miR-185-5p, miR-652-5p, and miR-1246 (from here on, referred as combo-miRs), into normal fibroblasts, effectively reprogramming them into cancer-associated fibroblasts. Here, we show that the conditioned medium from the fibroblasts activated by combo-miRs exerts a pro-tumorigenic effect on epithelial cells, enhancing the viability and migratory potential while driving increased invasiveness in patient-derived breast cancer organoids. A proteomic analysis of conditioned medium from combo-miRs activated fibroblasts revealed 76 significantly upregulated secreted proteins compared to control. Bioinformatic analysis identified the transcriptional factor PATZ1 as a potential regulator of the 12 most highly upregulated proteins. Consistently, in-silico predictions and in vitro experiments confirmed that PATZ1 is a direct target of miR-185-5p and miR-652-5p. The downregulation of PATZ1 by these miRNAs led to increased levels of the secreted proteins in the conditioned medium from combo-miRs activated fibroblasts. Furthermore, the conditioned medium from PATZ1-knockout mesenchymal embryonic fibroblasts and normal fibroblasts with silenced PATZ1 similarly enhanced the migratory potential of MCF10A cells, further supporting the critical role of PATZ1 in regulating tumor-promoting mechanisms. These findings provide valuable insights into the dynamics of the TME in TNBC, highlighting combo-miRs and PATZ1 as promising targets for future therapeutic interventions.
Ex.50.T aptamer impairs tumor–stroma cross-talk in breast cancer by targeting gremlin-1 Cristina Quintavalle, Francesco Ingenito, Giuseppina Roscigno, Birlipta Pattanayak, Carla Lucia Esposito, Alessandra Affinito, Danilo Fiore, Gianluca Petrillo, Silvia Nuzzo, Bartolomeo Della Ventura, Federica D’Aria, Concetta Giancola, Stefania Mitola, Elisabetta Grillo, Marinella Pirozzi, Greta Donati, Francesco Saverio Di Leva, Luciana Marinelli, Zoran Minic, Francesca De Micco, Guglielmo Thomas, Maxim V. Berezovski, Gerolama Condorelli Cell Death Discovery, 2025 The tumor microenvironment profoundly influences tumor complexity, particularly in breast cancer, where cancer-associated fibroblasts play pivotal roles in tumor progression and therapy resistance. Extracellular vesicles are involved in mediating communication within the TME, specifically highlighting their role in promoting the transformation of normal fibroblasts into cancer-associated fibroblasts. Recently, we identified an RNA aptamer, namely ex.50.T, that binds with remarkable affinity to extracellular vesicles shed from triple-negative breast cancer cells. Here, through in vitro assays and computational analyses, we demonstrate that the binding of ex.50.T to extracellular vesicles and parental breast cancer cells is mediated by recognition of gremlin-1 (GREM1), a bone morphogenic protein antagonist implicated in breast cancer aggressiveness and metastasis. Functionally, we uncover the role of ex.50.T as an innovative therapeutic agent in the process of tumor microenvironment re-modeling, impeding GREM1 signaling, blocking triple-negative breast cancer extracellular vesicles internalization in recipient cells, and counteracting the transformation of normal fibroblasts into cancer-associated fibroblasts. Altogether, our findings highlight ex.50.T as a novel therapeutical avenue for breast cancer and potentially other GREM1-dependent malignancies, offering insights into disrupting TME dynamics and enhancing cancer treatment strategies.
BH3 mimetic drugs overcome the microenvironment-induced resistance to crizotinib in ALK+ anaplastic large cell lymphoma Claudia Pignataro, Pietro Zoppoli, Luca Vincenzo Cappelli, Liron Yoffe, Marta Moretti, Mariapaola Izzo, Selene Mallia, Clarisse Kayembe, Abigail Taylor, Gianluca Petrillo, Alessandra Affinito, Cristina Quintavalle, Giada De Luca, Martina Mascolo, Sara Verde, Aurelia Fraticelli, Alessia Ciarrocchi, Paolo Salerno, Enrico De Smaele, Antonio Francesco Campese, Valentina Fragliasso, Robin Foà, Giuseppe Merla, Giorgio Inghirami, Gerolama Condorelli, Danilo Fiore Blood Advances, 2025 Resistance to first-line chemotherapies and crizotinib in anaplastic large cell lymphoma (ALCL) represents a significant challenge, often leading to a dismal outcome. Despite recent advancements, the dissection of the intrinsic and extrinsic molecular alterations underlying crizotinib resistance in ALCL is still poorly understood. Here, we transcriptionally unraveled the bidirectional interplay between anaplastic lymphoma kinase (ALK)-driven ALCL (ALK+ ALCL) and stromal cells in the presence of crizotinib at bulk and single-cell levels and identified that the microenvironment provides prosurvival signals leading to crizotinib persistence in ALK+ ALCL. We detected increased B-cell lymphoma 2 (BCL2) expression and downregulation of pathways related to apoptosis in crizotinib-persister ALK+ ALCL cells. Furthermore, we predicted in silico the ligand-receptor interactions between tumoral and stromal cells, supporting their contribution to ALCL pathogenesis mainly participating in the adhesion/membrane transport, triggering receptors, and promoting activation and microenvironment stimulation in lymphoma cells. Finally, we explored the effect of crizotinib in combination with BH3 mimetics. Pharmacologic and genetic ablation of anti-apoptotic targets displayed a significant synergistic effect with crizotinib, overcoming the stroma-mediated protection of lymphoma cells on drug treatment. Thus, BCL2/B-cell lymphoma-extra large (BCL-XL) targeting is synthetic lethal with crizotinib exposure in ALK+ ALCL and represents an intrinsic- and extrinsic-mediated targetable vulnerability in lymphoma cells challenged with crizotinib. Our data support the evaluation of BCL2 targeting in crizotinib-based regimens in the management of patients with ALK+ ALCL.
Preanalytical variables and analytes in liquid biopsy approach for brain tumors: A comprehensive review and recommendations from the RANO Group and the Brain Liquid Biopsy Consortium Chetan Bettegowda, Houtan Noushmehr, Alessandra Affinito, Manmeet S Ahluwalia, Olaf Ansorge, Katayoun Ayasoufi, Stephen Bagley, Jill Barnholtz-Sloan, Myron Best, Dieta Brandsma, Chaya Brodie, Anke Brüning-Richardson, Ana Valeria Castro, Susan M Chang, Gerolama Condorelli, Ahmad Daher, Vineet Datta, John de Groot, Pim French, Evanthia Galanis, Anna Golebiewska, Petra Hamerlik, C Oliver Hanemann, Matthias Holdhoff, Jason Huse, Mustafa Khasraw, Suzanne LeBlang, Beatrice Melin, Florent Mouliere, Claire O’Leary, Janusz Rak, Amitava Ray, Stephen Robinson, Ola Rominiyi, Federico Roncaroli, Roberta Rudà, Joan Seoane, Nik Sol, Martin J van den Bent, Michael A Vogelbaum, Tobias Walbert, Colin Watts, Tobias Weiss, Michael Weller, Patrick Y Wen, Victoria Wykes, Stephen Yip, Susan C Short, Riccardo Soffietti Neuro Oncology, 2025 This review explores the pivotal role of preanalytical variables in bringing liquid biopsy approaches into the clinic for brain tumors. Preanalytical variables encompass a range of critical issues, from blood sample collection and handling to the impact of tumor heterogeneity and patient-specific factors. These variables introduce challenges such as false positives, false negatives, and variability in the analysis of tumor signals, which can hinder the diagnostic and prognostic utility of liquid biopsies. Understanding the nuances of preanalytical variables is essential for the successful implementation of liquid biopsy in clinical settings. This paper delves into strategies aimed at mitigating the influence of preanalytical variables by emphasizing the importance of standardized sample collection protocols, optimized sample processing and storage, quality control measures, and the integration of multiple liquid biopsy modalities.
Targeting Glioblastoma Stem Cells via EphA2: Structural Insights into the RNA Aptamer A40s for Precision Therapy Isidora Diakogiannaki, Vincenzo Maria D’Amore, Alessandra Affinito, Greta Donati, Elpidio Cinquegrana, Cristina Quintavalle, Martina Mascolo, Jule Walter, Heike Betat, Mario Mörl, Francesco Saverio Di Leva, Gerolama Condorelli, Luciana Marinelli Journal of Chemical Information and Modeling, 2025 EphA2 receptor tyrosine kinase is overexpressed in many solid tumors and serves as a key driver of tumorigenesis and metastasis. It is highly expressed in glioblastoma multiforme, the most aggressive brain tumor in adults, and in its stem cells [glioblastoma stem cells (GSCs)], which contribute to treatment resistance and tumor relapse. In a previous study, we used the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) procedure, a method for selecting high-affinity nucleic acids to specific targets via iterative selection and amplification, to identify the 2'-fluorinated EphA2-targeting RNA aptamer A40L and a truncated 30-mer derivative, A40s. Both aptamers were able to inhibit GSC growth, stemness, and migration upon EphA2 binding. Here, by integrating computational and experimental methods, the A40s structure was unraveled and its interaction with EphA2 was investigated. Our model offers a blueprint to accelerate the development of optimized A40s variants, advancing next-generation EphA2-targeted anticancer therapies.
Targeting Glioblastoma Stem Cells: A40s Aptamer-NIR-Dye Conjugate for Glioblastoma Visualization and Treatment Alessandra Affinito, Francesco Ingenito, Sara Verde, Emanuele Musella, Birlipta Pattanayak, Danilo Fiore, Cristina Quintavalle, Aurelia Fraticelli, Martina Mascolo, Gianluca Petrillo, Claudia Pignataro, Giada De Luca, Laura Mezzanotte, Gerolama Condorelli Biomolecules, 2025 Glioblastoma (GBM) is the most aggressive and challenging brain cancer, in terms of diagnosis and therapy. The highly infiltrative glioblastoma stem cells (GSCs) are difficult to visualize and surgically remove with the current diagnostic tools, which often lead to misdiagnosis and false-positive results. In this study, we focused on a groundbreaking tool for specifically visualizing and removing GSCs. We exploited the specific binding of A40s aptamer to EphA2 for the selective delivery of Near-Infrared Dyes (NIR-Dyes), like IR700DX and ICG, both in vitro and in vivo. The A40s aptamer, engineered through the NIR-Dye conjugation, did not affect aptamer binding ability; indeed, A40s-NIR-Dye conjugates bound GLI261 stem-like cells and patient-derived GSCs in vitro; moreover, they induced cell death upon photodynamic therapy treatment (PDT). Additionally, when systemically administrated, the A40s-NIR-Dye conjugates allowed GSC visualization and accumulated in tumor mass. This allows GSCs detection and treatment. Our findings demonstrate the potential use of A40s aptamer as a targeted therapeutic approach and imaging tool in vivo for GSCs, paving the way for improved, more effective, and less invasive GBM management.
MCT4-driven CAF-mediated metabolic reprogramming in breast cancer microenvironment is a vulnerability targetable by miR-425-5p Alessandra Affinito, Cristina Quintavalle, Rosario Vincenzo Chianese, Giuseppina Roscigno, Danilo Fiore, Valeria D’Argenio, Guglielmo Thomas, Alessia Savarese, Francesco Ingenito, Lorenza Cocca, Silvia Nuzzo, Maxim V. Berezovski, Maria Patrizia Stoppelli, Gerolama Condorelli Cell Death Discovery, 2024 Multiple oncogenic alterations contribute to breast cancer development. Metabolic reprogramming, deeply contributing to tumor microenvironment (TME) education, is now widely recognized as a hallmark of cancer. The reverse Warburg effect induces cancer-associated fibroblasts (CAFs) to produce and secrete L-lactate, enhancing malignant characteristics such as neoangiogenesis, metastatic dissemination, and treatment resistance. Monocarboxylate transporter (MCT) 4 is involved in lactate efflux from CAFs into stromal and epithelial cells. Here, we first assess the expression of miR-425-5p and its target MCT4 in breast cancer CAFs and normal fibroblasts. We analyzed the metabolic changes induced by miR-425-5p in CAFs and its role in the education of breast cancer epithelial cells. We show that miR-425-5p-induced MCT4 knockdown decreased lactate extrusion from CAFs and its availability in the TME. miR-425-5p overexpression induced profound metabolic transformation in CAFs, ultimately influencing breast cancer metabolism. Furthermore, miR-425-5p impaired the capacity of CAFs to sustain vessel formation and breast cancer cell migration, viability, and proliferation. These findings emphasize the key role of miR-425-5p in breast cancer metabolism and aggressiveness, and its possible importance for breast cancer therapy and monitoring.
EFFECTS OF DRUGS ON THYROID FUNCTION Gianfranco Fenzi, Mario Vitale, Vincenzo Bassi, Gerolama Condorelli Thyroid Diseases Clinical Fundamentals and Therapy, 2024
One-Month Dual Antiplatelet Therapy After Bioresorbable Polymer Everolimus-Eluting Stents in High Bleeding Risk Patients Carlo A. Pivato, Bernhard Reimers, Luca Testa, Andrea Pacchioni, Carlo Briguori, Carmine Musto, Giovanni Esposito, Raffaele Piccolo, Luigi Lucisano, Leonardo De Luca, Federico Conrotto, Andrea De Marco, Anna Franzone, Patrizia Presbitero, Giuseppe Ferrante, Gerolama Condorelli, Valeria Paradies, Gennaro Sardella, Ciro Indolfi, Gianluigi Condorelli, Giulio G. Stefanini Journal of the American Heart Association, 2022
Structure- and Interaction-Based Design of Anti-SARS-CoV-2 Aptamers Vladimir Mironov, Irina A. Shchugoreva, Polina V. Artyushenko, Dmitry Morozov, Nicola Borbone, Giorgia Oliviero, Tatiana N. Zamay, Roman V. Moryachkov, Olga S. Kolovskaya, Kirill A. Lukyanenko, Yanling Song, Iuliia A. Merkuleva, Vladimir N. Zabluda, Georgy Peters, Lyudmila S. Koroleva, Dmitry V. Veprintsev, Yury E. Glazyrin, Ekaterina A. Volosnikova, Svetlana V. Belenkaya, Tatiana I. Esina, Anastasiya A. Isaeva, Valentina S. Nesmeyanova, Daniil V. Shanshin, Anna N. Berlina, Nadezhda S. Komova, Valery A. Svetlichnyi, Vladimir N. Silnikov, Dmitriy N. Shcherbakov, Galina S. Zamay, Sergey S. Zamay, Tatyana Smolyarova, Elena P. Tikhonova, Kelvin H.‐C. Chen, U‐Ser Jeng, Gerolama Condorelli, Vittorio Franciscis, Gerrit Groenhof, Chaoyong Yang, Alexander A. Moskovsky, Dmitri G. Fedorov, Felix N. Tomilin, Weihong Tan, Yuri Alexeev, Maxim V. Berezovski, Anna S. Kichkailo Chemistry A European Journal, 2022
Interfering with the Tumor-Immune Interface: Making Way for Triazine-Based Small Molecules as Novel PD-L1 Inhibitors Pasquale Russomanno, Giulia Assoni, Jussara Amato, Vincenzo Maria D’Amore, Riccardo Scaglia, Diego Brancaccio, Martina Pedrini, Giovanna Polcaro, Valeria La Pietra, Paolo Orlando, Marianna Falzoni, Linda Cerofolini, Stefano Giuntini, Marco Fragai, Bruno Pagano, Greta Donati, Ettore Novellino, Cristina Quintavalle, Gerolama Condorelli, Francesco Sabbatino, Pierfausto Seneci, Daniela Arosio, Stefano Pepe, Luciana Marinelli Journal of Medicinal Chemistry, 2021
Urinary Dickkopf-3 and Contrast-Associated Kidney Damage Giuseppina Roscigno, Cristina Quintavalle, Giuseppe Biondi-Zoccai, Francesca De Micco, Giacomo Frati, Alessandra Affinito, Silvia Nuzzo, Gerolama Condorelli, Carlo Briguori Journal of the American College of Cardiology, 2021
STAT3 targeting by an aptamer-based conjugate for glioblastoma multiforme therapy C.L. Esposito, , S Nuzzo, M Ibba, I Grinev, A Gorbushin, D Grek, I Voronkovskii, O Kolovskaya, T Zamay, E Morozov, A Koshmanova, A Narodov, V Khorzhevskii, E Erakhtin, A Krat, A Yakovlev, P Shesternya, A Kichkailo, G Condorelli, S Catuogno, V Franciscis, , , , , , , , , , , , , , , , , , , and Siberian Medical Review, 2021
Ranolazine protects from doxorubicin-induced oxidative stress and cardiac dysfunction Carlo G. Tocchetti, Andrea Carpi, Carmela Coppola, Cristina Quintavalle, Domenica Rea, Marika Campesan, Antonella Arcari, Giovanna Piscopo, Clemente Cipresso, Maria Gaia Monti, Claudia De Lorenzo, Claudio Arra, Gerolama Condorelli, Fabio Di Lisa, Nicola Maurea European Journal of Heart Failure, 2014
Multifunctional aptamer-miRNA conjugates for targeted cancer therapy Carla L Esposito, Laura Cerchia, Silvia Catuogno, Gennaro De Vita, Justin P Dassie, Gianluca Santamaria, Piotr Swiderski, Gerolama Condorelli, Paloma H Giangrande, Vittorio de Franciscis Molecular Therapy, 2014
Assessment of the 9p21.3 locus in severity of coronary artery disease in the presence and absence of type 2 diabetes Natalia V Rivera, Robert Carreras-Torres, Roberta Roncarati, Chiara Viviani-Anselmi, Francesca De Micco, Alessandra Mezzelani, Werner Koch, Petra Hoppmann, Adnan Kastrati, Alexandre FR Stewart, Li Chen, Robert Roberts, Lennart C Karssen, Najaf Amin, Valentina Trimarco, Raffaele Izzo, Guido Iaccarino, Gerolama Condorelli, Annibale A Puca, Paolo Pagnotta, Flavio Airoldi, Bruno Trimarco, Cornelia M van Duijn, Gianluigi Condorelli, Carlo Briguori BMC Medical Genetics, 2013
Electrochemical detection of miRNA-222 by use of a magnetic bead-based bioassay Francesca Bettazzi, Ezat Hamid-Asl, Carla Lucia Esposito, Cristina Quintavalle, Nello Formisano, Serena Laschi, Silvia Catuogno, Margherita Iaboni, Giovanna Marrazza, Marco Mascini, Laura Cerchia, Vittorio De Franciscis, Gerolama Condorelli, Ilaria Palchetti Analytical and Bioanalytical Chemistry, 2013
Impact of a high loading dose of atorvastatin on contrast-induced acute kidney injury Cristina Quintavalle, Danilo Fiore, Francesca De Micco, Gabriella Visconti, Amelia Focaccio, Bruno Golia, Bruno Ricciardelli, Elvira Donnarumma, Antonio Bianco, Maria Assunta Zabatta, Giancarlo Troncone, Antonio Colombo, Carlo Briguori, Gerolama Condorelli Circulation, 2012
Epigenetic Regulation of miR-212 Expression in Lung Cancer Mariarosaria Incoronato, Loredana Urso, Ana Portela, Mikko O. Laukkanen, Ylermi Soini, Cristina Quintavalle, Simona Keller, Manel Esteller, Gerolama Condorelli Plos One, 2011
PED is overexpressed and mediates TRAIL resistance in human non-small cell lung cancer Ciro Zanca, Michela Garofalo, Cristina Quintavalle, Giulia Romano, Mario Acunzo, Pia Ragno, Nunzia Montuori, Mariarosaria Incoronato, Luigi Tornillo, Daniel Baumhoer, Carlo Briguori, Luigi Terracciano, Gerolama Condorelli Journal of Cellular and Molecular Medicine, 2008
Induction and intracellular regulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) mediated apotosis in human malignant glioma cells Cancer Research, 2001
Thyrotropin regulates autophosphorylation and kinase activity of both the insulin and the insulin-like growth factor-I receptors in FRTL5 cells Endocrinology, 1992
Antiphosphotyrosine immunoprecipitation of an insulin-stimulated receptor phosphatase activity from FRTL5 cells Endocrinology, 1991
Insulin and insulin-like growth factor I (IGF I) stimulate phosphorylation of a M(r) 175,000 cytoskeleton-associated protein in intact FRTL5 cells Journal of Biological Chemistry, 1989
