Rita Lombardi

Scopus Publications

Dual inhibition of mTOR and HSP90 enhances cisplatin efficacy and overcomes resistance in ovarian cancer
Rita Lombardi, Laura Addi, Biagio Pucci, Francesca Bruzzese, Maura Sonego, et al.
Cell Death and Disease, 2026
Epithelial ovarian cancer (EOC) represents the most lethal gynecological disease, with a 5-year relative survival rate of 46% after the diagnosis. Standard treatment includes surgery followed by platinum (Pt)-based chemotherapy. However, Pt-resistance frequently occurs and strongly impact on the survival of EOC patients for whom we still do not have valid therapeutic options. By using a proteomic approach, we previously demonstrated a potential role of HSP90 in the mechanism of resistance in vitro, ex vivo e partially in vivo. To further investigate in depth the mechanism by which EOC cells acquired Pt-resistance, we used a quantitative phosphoproteomics approach followed by enrichment functional analysis. Here, we identified 542 differentially expressed phosphoproteins in Pt-resistant compared to parental cells identifying mTOR and HSF1 as the most enriched pathways. The up-regulation of the phosphorylated form of PDK1, AKT, mTOR, and RPS6 was observed in Pt-resistant compared to parental cells. Moreover, we also demonstrated the up-regulation of the activity of HSF1 along with the elevation crucial components of the chaperone complex machinery HSP90, HSP70 and HSP40. Since mTOR is an attractive target for therapeutic intervention because of its key role in the crosstalk of various signaling pathways, we propose a novel therapeutic strategy based on the pharmacologic inhibition of HSP90 and mTOR able to further potentiate the Pt-based chemotherapy. Accordingly, the combination of ganetespib (an HSP90 inhibitor) and temsirolimus (a FDA approved-mTOR inhibitor) with cisplatin synergistically reduced colony formation and microtissues cell growth in vitro by increasing DNA-damage and apoptosis and in vivo enhancing mouse survival. Mechanistically, the triple combination treatment, impaired the proteins involved in mTOR signaling and HSF1 transactivation. Notably, all these data were confirmed also in Pt-resistant Non Small Cell Lung Cancer models. Collectively, our findings identify a promising new antitumor strategy for the treatment of Pt-resistance in cancer patients.
Valproic acid improves the efficacy of oxaliplatin/fluoropyrimidine-based chemotherapy by targeting cancer stem cell via β-Catenin modulation in colorectal cancer
Maria Serena Roca, Rita Lombardi, Cristina Testa, Federica Iannelli, Laura Grumetti, et al.
Cell Death and Disease, 2025
Despite advances in systemic therapeutic approaches, metastatic colorectal cancer (mCRC) patients harboring BRAF or RAS mutations have poor outcomes. Cancer stem cells (CSCs) play central roles in drug resistance and CRC recurrence. Therefore, targeting the epigenetic mechanisms that sustain CSC properties is a promising therapeutic approach. In this study, we report the efficacy of a treatment strategy with the potential to overcome chemotherapy resistance that involves administering the well-known antiepileptic drug and epigenetic agent valproic acid (VPA) and the standard chemotherapy regimen of oxaliplatin/fluoropyrimidine to wild-type CSCs and CSCs with BRAF and RAS mutations in enriched primary spheroid cultures. Notably, we demonstrated that VPA plus chemotherapy was more effective than other epigenetic drug-chemotherapy combinations by inhibiting cell proliferation and clonogenic growth and by inducing apoptosis and DNA damage. Mechanistically, proteomic analysis demonstrated that VPA induced CSC differentiation through the critical target of VPA, β-Catenin. Indeed, VPA promoted the proteasome-dependent degradation of β-Catenin by enhancing its binding to the E2 ubiquitin-conjugating enzyme UBE2a, leading to marked reductions in nuclear and cytoplasmic β-Catenin levels and subsequently decreasing β-Catenin/TCF-LEF target promoter activation. These effects were confirmed in three in vivo CRC xenograft models, including a syngeneic CT26 immunocompetent mouse model, where VPA combined with oxaliplatin/capecitabine chemotherapy and anti-VEGF therapy, a standard first-line treatment for mCRC, significantly suppressed tumor growth and prolonged survival with minimal toxicity. Proteomic analysis of tumor tissues from in vivo CRC models confirmed the VPA-mediated downregulation of CSC markers and β-Catenin.
Serum-Based Proteomic Approach to Identify Clinical Biomarkers of Radiation Exposure
Emeshaw Damtew Zebene, Biagio Pucci, Rita Lombardi, Hagos Tesfay Medhin, Edom Seife, et al.
Cancers, 2025
Background: Ionizing radiation (IR) exposure poses a significant health risk due to its widespread use in medical diagnostics and therapeutic applications, necessitating rapid and effective biomarkers for assessment. Objective: The aim of this study is to identify the serum proteomic signature of IR exposure in patients undergoing radiotherapy (RT). Methods: Blood samples were obtained from eighteen patients with head and neck cancer (HNC) and five patients with rectal cancer before and immediately after they underwent curative intensity-modulated radiotherapy (IMRT). The comprehensive serum proteome was analyzed in individual samples using nanoHPLC-MS/MS. Results: Forty radiation-modulated proteins (RMPs), 24 upregulated and 16 downregulated, with a fold change ≥1.5 and p-value < 0.05 were identified. About 40% of the RMPs are involved in acute phase response, DNA repair, and inflammation; the key RMPs were ADCY1, HGF, MCEMP1, CHD4, RECQL5, MSH6, and ZNF224. Conclusions: This study identifies a panel of serum proteins that may reflect the radiation response, providing a valuable molecular fingerprint of IR exposure and paving the way for the development of sensitive and specific biomarkers for early detection and clinical management of IR-related injuries.
Proteomic Analysis of Biomarkers Predicting Treatment Response in Patients with Head and Neck Cancers
Emeshaw Damtew Zebene, Rita Lombardi, Biagio Pucci, Hagos Tesfay Medhin, Edom Seife, et al.
International Journal of Molecular Sciences, 2024
Head and neck cancers (HNCs) are the sixth most commonly diagnosed cancer and the eighth leading cause of cancer-related mortality worldwide, with squamous cell carcinoma being the most prevalent type. The global incidence of HNCs is steadily increasing, projected to rise by approximately 30% per year by 2030, a trend observed in both developed and undeveloped countries. This study involved serum proteomic profiling to identify predictive clinical biomarkers in cancer patients undergoing chemoradiotherapy (CRT). Fifteen HNC patients at Tikur Anbessa Specialized Hospital, Radiotherapy (RT) center in Addis Ababa were enrolled. Serum samples were collected before and after RT, and patients were classified as responders (R) or non-responders (NR). Protein concentrations in the serum were determined using the Bradford assay, followed by nano-HPLC–MS/MS for protein profiling. Progenesis QI for proteomics identified 55 differentially expressed proteins (DEPs) between R and NR, with a significance of p < 0.05 and a fold-change (FC) ≥ 1.5. The top five-up-regulated proteins included MAD1L1, PSMC2, TRIM29, C5, and SERPING1, while the top five-down-regulated proteins were RYR1, HEY2, HIF1A, TF, and CNN3. Notably, about 16.4% of the DEPs were involved in cellular responses to DNA damage from cancer treatments, encompassing proteins related to deoxyribonucleic acid (DNA) damage sensing, checkpoint activation, DNA repair, and apoptosis/cell cycle regulation. The analysis of the relative abundance of ten proteins with high confidence scores identified three DEPs: ADIPOQ, HEY2, and FUT10 as potential predictive biomarkers for treatment response. This study highlighted the identification of three potential predictive biomarkers—ADIPOQ, HEY2, and FUT10—through serum proteomic profiling in HNC patients undergoing RT, emphasizing their significance in predicting treatment response.
Integrated proteomics and metabolomics analyses reveal new insights into the antitumor effects of valproic acid plus simvastatin combination in a prostate cancer xenograft model associated with downmodulation of YAP/TAZ signaling
Federica Iannelli, Rita Lombardi, Susan Costantini, Maria Serena Roca, Laura Addi, et al.
Cancer Cell International, 2024
Inhibition of autophagy by chloroquine prevents resistance to PI3K/AKT inhibitors and potentiates their antitumor effect in combination with paclitaxel in triple negative breast cancer models
Stefania Cocco, Alessandra Leone, Maria Serena Roca, Rita Lombardi, Michela Piezzo, et al.
Journal of Translational Medicine, 2022
Background Triple negative breast cancer (TNBC) is an aggressive disease characterized by high risk of relapse and development of resistance to different chemotherapy agents. Several targeted therapies have been investigated in TNBC with modest results in clinical trials. Among these, PI3K/AKT inhibitors have been evaluated in addition to standard therapies, yielding conflicting results and making attempts on elucidating inherent mechanisms of resistance of great interest. Increasing evidences suggest that PI3K/AKT inhibitors can induce autophagy in different cancers. Autophagy represents a supposed mechanism of drug-resistance in aggressive tumors, like TNBC. We, therefore, investigated if two PI3K/AKT inhibitors, ipatasertib and taselisib, could induce autophagy in breast cancer models, and whether chloroquine (CQ), a well known autophagy inhibitor, could potentiate ipatasertib and taselisib anti-cancer effect in combination with conventional chemotherapy. Methods The induction of autophagy after ipatasertib and taselisib treatment was evaluated in MDAMB231, MDAM468, MCF7, SKBR3 and MDAB361 breast cancer cell lines by assaying LC3-I conversion to LC3-II through immunoblotting and immunofluorescence. Other autophagy-markers as p62/SQSTM1 and ATG5 were evaluated by immunoblotting. Synergistic antiproliferative effect of double and triple combinations of ipatasertib/taselisib plus CQ and/or paclitaxel were evaluated by SRB assay and clonogenic assay. Anti-apoptotic effect of double combination of ipatasertib/taselisib plus CQ was evaluated by increased cleaved-PARP by immunoblot and by Annexin V- flow cytometric analysis. In vivo experiments were performed on xenograft model of MDAMB231 in NOD/SCID mice. Results Our results suggested that ipatasertib and taselisib induce increased autophagy signaling in different breast cancer models. This effect was particularly evident in PI3K/AKT resistant TNBC cells, where the inhibition of autophagy by CQ potentiates the therapeutic effect of PI3K/AKT inhibitors in vitro and in vivo TNBC models, synergizing with taxane-based chemotherapy. Conclusion These data suggest that inhibition of authophagy with CQ could overcome mechanism of drug resistance to PI3K/AKT inhibitors plus paclitaxel in TNBC making the evaluation of such combinations in clinical trials warranted.
Epigenetic Approaches to Overcome Fluoropyrimidines Resistance in Solid Tumors
Laura Grumetti, Rita Lombardi, Federica Iannelli, Biagio Pucci, Antonio Avallone, et al.
Cancers, 2022
Although fluoropyrimidines were introduced as anticancer agents over 60 years ago, they are still the backbone of many combination chemotherapy regimens for the treatment of solid cancers. Like other chemotherapeutic agents, the therapeutic efficacy of fluoropyrimidines can be affected by drug resistance and severe toxicities; thus, novel therapeutic approaches are required to potentiate their efficacy and overcome drug resistance. In the last 20 years, the deregulation of epigenetic mechanisms has been shown to contribute to cancer hallmarks. Histone modifications play an important role in directing the transcriptional machinery and therefore represent interesting druggable targets. In this review, we focused on histone deacetylase inhibitors (HDACis) that can increase antitumor efficacy and overcome resistance to fluoropyrimidines by targeting specific genes or proteins. Our preclinical data showed a strong synergistic interaction between HDACi and fluoropyrimidines in different cancer models, but the clinical studies did not seem to confirm these observations. Most likely, the introduction of increasingly complex preclinical models, both in vitro and in vivo, cannot recapitulate human complexity; however, our analysis of clinical studies revealed that most of them were designed without a mechanistic approach and, importantly, without careful patient selection.
An Integrated In Silico, In Vitro and Tumor Tissues Study Identified Selenoprotein S (SELENOS) and Valosin-Containing Protein (VCP/p97) as Novel Potential Associated Prognostic Biomarkers in Triple Negative Breast Cancer
Susan Costantini, Andrea Polo, Francesca Capone, Marina Accardo, Angela Sorice, et al.
Cancers, 2022
Background. Triple negative breast cancer (TNBC) is a heterogeneous group of tumors with early relapse, poor overall survival, and lack of effective treatments. Hence, new prognostic biomarkers and therapeutic targets are needed. Methods. The expression profile of all twenty-five human selenoproteins was analyzed in TNBC by a systematic approach.In silicoanalysis was performed on publicly available mRNA expression datasets (Cancer Cell Line Encyclopedia, CCLE and Library of Integrated Network-based Cellular Signatures, LINCS). Reverse transcription quantitative PCR analysis evaluated selenoprotein mRNA expression in TNBC versus non-TNBC and normal breast cells, and in TNBC tissues versus normal counterparts. Immunohistochemistry was employed to study selenoproteins in TNBC tissues. STRING and Cytoscape tools were used for functional and network analysis. Results.GPX1, GPX4, SELENOS, TXNRD1 and TXNRD3 were specifically overexpressed in TNBC cells, tissues and CCLE/LINCS datasets. Network analysis demonstrated that SELENOS-binding valosin-containing protein (VCP/p97) played a critical hub role in the TNBCselenoproteins sub-network, being directly associated with SELENOS expression. The combined overexpression of SELENOS and VCP/p97 correlated with advanced stages and poor prognosis in TNBC tissues and the TCGA dataset. Conclusion. Combined evaluation of SELENOS and VCP/p97 might represent a novel potential prognostic signature and a therapeutic target to be exploited in TNBC.
HSP90 identified by a proteomic approach as druggable target to reverse platinum resistance in ovarian cancer
Rita Lombardi, Maura Sonego, Biagio Pucci, Laura Addi, Federica Iannelli, et al.
Molecular Oncology, 2021
Acquired resistance to platinum (Pt)‐based therapies is an urgent unmet need in the management of epithelial ovarian cancer (EOC) patients. Here, we characterized by an unbiased proteomics method three isogenic EOC models of acquired Pt resistance (TOV‐112D, OVSAHO, and MDAH‐2774). Using this approach, we identified several differentially expressed proteins in Pt‐resistant (Pt‐res) compared to parental cells and the chaperone HSP90 as a central hub of these protein networks. Accordingly, up‐regulation of HSP90 was observed in all Pt‐res cells and heat‐shock protein 90 alpha isoform knockout resensitizes Pt‐res cells to cisplatin (CDDP) treatment. Moreover, pharmacological HSP90 inhibition using two different inhibitors [17‐(allylamino)‐17‐demethoxygeldanamycin (17AAG) and ganetespib] synergizes with CDDP in killing Pt‐res cells in all tested models. Mechanistically, genetic or pharmacological HSP90 inhibition plus CDDP ‐induced apoptosis and increased DNA damage, particularly in Pt‐res cells. Importantly, the antitumor activities of HSP90 inhibitors (HSP90i) were confirmed both ex vivo in primary cultures derived from Pt‐res EOC patients ascites and in vivo in a xenograft model. Collectively, our data suggest an innovative antitumor strategy, based on Pt compounds plus HSP90i, to rechallenge Pt‐res EOC patients that might warrant further clinical evaluation.
Synergistic antitumor interaction of valproic acid and simvastatin sensitizes prostate cancer to docetaxel by targeting CSCs compartment via YAP inhibition
Federica Iannelli, Maria Serena Roca, Rita Lombardi, Chiara Ciardiello, Laura Grumetti, et al.
Journal of Experimental and Clinical Cancer Research, 2020
Background Despite the introduction of several novel therapeutic approaches that improved survival, metastatic castration-resistant prostate cancer (mCRPC) remains an incurable disease. Herein we report the synergistic antitumor interaction between two well-known drugs used for years in clinical practice, the antiepileptic agent with histone deacetylase inhibitory activity valproic acid and the cholesterol lowering agent simvastatin, in mCRPC models. Methods Synergistic anti-tumor effect was assessed on PC3, 22Rv1, DU145, DU145R80, LNCaP prostate cancer cell lines and EPN normal prostate epithelial cells, by calculating combination index (CI), caspase 3/7 activation and colony formation assays as well as on tumor spheroids and microtissues scored with luminescence 3D-cell viability assay. Cancer stem cells (CSC) compartment was studied evaluating specific markers by RT-PCR, western blotting and flow cytometry as well as by limiting dilution assay. Cholesterol content was evaluated by 1H-NMR. Overexpression of wild-type YAP and constitutively active YAP5SA were obtained by lipofectamine-based transfection and evaluated by immunofluorescence, western blotting and RT-PCR. 22Rv1 R_39 docetaxel resistant cells were selected by stepwise exposure to increasing drug concentrations. In vivo experiments were performed on xenograft models of DU145R80, 22Rv1 parental and docetaxel resistant cells, in athymic mice. Results We demonstrated the capacity of the combined approach to target CSC compartment by a novel molecular mechanism based on the inhibition of YAP oncogene via concurrent modulation of mevalonate pathway and AMPK. Because both CSCs and YAP activation have been associated with chemo-resistance, we tested if the combined approach can potentiate docetaxel, a standard of care in mCRCP treatment. Indeed, we demonstrated, both in vitro and in vivo models, the ability of valproic acid/simvastatin combination to sensitize mCRPC cells to docetaxel and to revert docetaxel-resistance, by mevalonate pathway/YAP axis modulation. Conclusion Overall, mCRPC progression and therapeutic resistance driven by CSCs via YAP, can be tackled by the combined repurposing of two generic and safe drugs, an approach that warrants further clinical development in this disease.
Valproic Acid Synergizes With Cisplatin and Cetuximab in vitro and in vivo in Head and Neck Cancer by Targeting the Mechanisms of Resistance
Federica Iannelli, Andrea Ilaria Zotti, Maria Serena Roca, Laura Grumetti, Rita Lombardi, et al.
Frontiers in Cell and Developmental Biology, 2020
Novel pathways involved in cisplatin resistance identified by a proteomics approach in non-small-cell lung cancer cells
Maria Rita Milone, Rita Lombardi, Maria Serena Roca, Francesca Bruzzese, Laura Addi, et al.
Journal of Cellular Physiology, 2019
Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation
Chiara Ciardiello, Alessandra Leone, Paola Lanuti, Maria S. Roca, Tania Moccia, et al.
Journal of Experimental and Clinical Cancer Research, 2019
Targeting mevalonate pathway in cancer treatment: Repurposing of statins
Federica Iannelli, Rita Lombardi, Maria R. Milone, Biagio Pucci, Simona De Rienzo, et al.
Recent Patents on Anti Cancer Drug Discovery, 2018
Proteomic characterization of peroxisome proliferator-activated receptor-γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype
Maria Rita Milone, Biagio Pucci, Tommaso Colangelo, Rita Lombardi, Federica Iannelli, et al.
Molecular Oncology, 2016
Proteomic analysis of zoledronic-acid resistant prostate cancer cells unveils novel pathways characterizing an invasive phenotype
Oncotarget, 2015
Annexin A1 is involved in the acquisition and maintenance of a stem cell-like/aggressive phenotype in prostate cancer cells with acquired resistance to zoledronic acid
Valentina Bizzarro, Raffaella Belvedere, Maria Rita Milone, Biagio Pucci, Rita Lombardi, et al.
Oncotarget, 2015