Scopus Publications
- A metabolic trade-off between malignant plasma cells and mesenchymal stromal cells sustains multiple myeloma growth
Giuseppe Taurino, Erika Griffini, Denise Toscani, Chiara Maccari, Saverio Tardito, et al.
Blood Neoplasia, 2026 - TARGETING GLUTAMINE METABOLISM POTENTIATES T CELL ENGAGER–MEDIATED IMMUNOTHERAPY IN MULTIPLE MYELOMA
Vincenzo Raimondi
Haematologica, 2026
Introduction. Despite the introduction of novel immunotherapeutic agents, such as CAR-T cells and T cell engagers (TCE), multiple myeloma (MM) remains an incurable disease with frequent relapse and immune evasion. MM cells exhibit a marked dependence on extracellular glutamine (Gln) to sustain anabolic growth, survival, and redox balance, due to the absence of Gln synthetase and high glutaminase (GLS) expression. This reliance establishes a competitive metabolic microenvironment within the bone marrow (BM) niche, where plasma cells (PCs) deplete Gln and limit its availability to immune cells. Conversely, activated T cells, although Gln-dependent, retain metabolic flexibility and can use alternative substrates under Gln restriction. We aimed to explore how altered Gln metabolism impacts immunotherapy response to BCMA/CD3 TCE. We hypothesized that targeting Gln metabolism may increase MM susceptibility to T cell-mediated cytotoxicity while maintaining T cell effector function.Methods. We investigated the effects of Gln restriction or pharmacological modulation on elranatamab activity, a clinically available BCMA/CD3 TCE. BCMA expression was assessed in human myeloma cell lines (HMCLs) and CD138⁺ PCs from 29 MM patients, via flow cytometry. Cytotoxicity and T cell activation (CD69, CD25) were assessed both in co-cultures of BCMA⁺ HMCLs with healthy donor T cells (E:T = 5:1) and in BM mononuclear cells (MNCs) from MM patients. Gln restriction was achieved by culturing HMCLs and BM MNCs in media containing 0.5 mM and 0.2 mM Gln, respectively, for 48 and 72 hours. GLS inhibition was tested using the CB-839 at 0.5 µM for HMCLs and 10 µM for BM MNCs.Results. Elranatamab elicited potent and antigen-specific cytotoxicity against HMCLs and primary MM cells, accompanied by robust T cell activation. Gln restriction did not reduce TCE-mediated cytotoxicity against HMCLs, which remained comparable to Gln standard conditions. Concurrently, T cell activation was unaffected, indicating a preserved effector functionality. Combined treatment with elranatamab and CB-839 significantly enhanced anti-MM cytotoxic activity compared with either agent alone, without impairing T cell activation. Extending to a more complex ex vivo setting, BM MNCs from six MM patients showed that Gln restriction significantly enhanced elranatamab-mediated cytotoxicity, while T cell activation remained preserved or even increased. Notably, all patient-derived samples responded more effectively to elranatamab under Gln-limited conditions, including those poorly or non-responsive in Gln standard medium. Finally, preliminary data indicate that combining elranatamab with CB-839 further enhances tumor cell lysis of primary MM cells, recapitulating the cell line findings.Conclusions. Collectively, these data suggest a novel rationale to integrate GLS inhibition with BCMA/CD3 TCE as a strategy to overcome metabolic immunosuppression and enhance immunotherapeutic responses in MM patients. - Identification By A Single Cell Analysis Of Ccn5/Wisp2 As A Possible Marker Of The Osteogenic Impairment In The Myeloma Bone Microenvironment
Camilla Sitzia
Haematologica, 2026
Introduction: In multiple myeloma (MM), bone disease arises from an imbalance between bone formation and resorption, leading to osteolytic damage and the establishment of a tumour-permissive bone microenvironment (BME). Evidence suggest that an increase of bone resorption process occurs in monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM), whereas the disruption of the osteoblastic niche characterizes the progression to MM. Recently our group using a single-cell atlas of osteoblastic lineage cells have begun to describe the BME of MM patients and its precursors disease to identify possible markers of bone disease and tumoral progression. Thus, in this study we investigated the possible role of CCN5/Wisp2, a regulator of Wnt and TGF-β signalling, in the osteogenic differentiation process and in osteoblast dysfunction in MM as compared to precursors diseases.Methods: Bone cells from 16 bone biopsies from MGUS, SMM, or newly diagnosed MM patients were analysed by single-cell RNA sequencing. For in vitro validation, hTERT-MSCs and 17 primary MSCs (SMM=8 and MM=9) were differentiated to OBs for 21 days with ascorbic acid and dexamethasone. Osteogenic commitment and CCN5 expression were assessed by qPCR. To test the influence of MM-derived factors, differentiating cultures were exposed to conditioned media (CM) from human myeloma cell lines (HMCLs).Results: scRNA-seq profiling of 42,823 BME cells identified 14 transcriptionally distinct to mesenchymal to OBs clusters. CCN5 expression increased progressively along the osteogenic differentiation, from proliferating/immature MSCs and peaking in pre-OBs, OBs, and also in one adipocyte-like cluster. Stage-wise comparison revealed significantly higher CCN5 expression in MGUS than in MM pre-OBs and OBs, suggesting an early activation of this pathway that decreases with disease progression. Notably, pre-OBs and OBs from non-progressor SMM patients displayed significantly higher CCN5 levels compared to those from SMM-progressors. In vitro CCN5 was strongly upregulated during osteogenic differentiation of hTERT-MSCs, with expression peaking at days 7 and 14. In primary patient-derived MSCs, CCN5 expression was significantly upregulated at day 21 in SMM-derived, but not in MM-derived OBs. Exposure to CM from 2 HMCLs (JJN3 and OPM2) significant downregulated CCN5 in hTERT-derived and SMM-derived OBs, whereas MM-derived OBs were largely unresponsive. Ongoing work includes immunohistochemical localization of CCN5 in bone biopsies and functional modulation through viral-mediated knockdown or overexpression.Conclusions: By integrating single-cell and functional analyses, our data suggest that CCN5 upregulation could characterize an osteogenic program preserved in MGUS and SMM non-progressors but progressively suppressed by myeloma-derived factors. Overall, these data identify CCN5 gene as a potential biomarker of the osteogenic impairment that occurs in MM patients. - FROM PRECURSOR DISEASES TO MULTIPLE MYELOMA: REMODELING OF THE OSTEOBLASTIC NICHE AT SINGLE CELL RESOLUTION
Mattia Dessena
Haematologica, 2026
Introduction: Disrupted osteoblastogenesis and impaired osteoblast (OB) functionality, driven by malignant plasma cells within the bone marrow, characterize multiple myeloma (MM) and its precursor diseases as monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM). In this context OB niche exhibit transcriptional and functional changes that may promote tumor progression. We aimed to characterize the OB population dynamics at single-cell resolution to identify alterations involved in MGUS to MM progression.Methods: Rare non-hematopoietic bone microenvironment (BME) cells from 16 patient biopsies from MGUS, SMM, and MM were analyzed by scRNAseq depleting CD235a+, CD45+, CD31+, and CD138+ cells using Chromium 10X Genomics platform. Scanpy version 1.10.4 package was used to identify differentially expressed genes. Pathway and biological process (BPs) enrichment were inferred via ORA and GSVA, while cell trajectories and interactions were assessed using STREAM and CellPhoneDB. Results: A total of 9,565 osteoblastic cells were profiled revealing three clusters: two representing pre-OB states with distinct functional features that segregate along bifurcating branches of the pseudotime-inferred trajectory. The first pre-OB cluster(Wisp2+) exhibited elevated expression of osteoblastogenesis-associated genes and hematopoietic supportive factors, together with a increased expression of BPs related to extracellular matrix mineralization. The second, pre OBs, showed a more immunosuppressive profile, attenuated modulation of bone growth and osteoblastic differentiation, and the expression of dysfunctional genes. The OBs represented phenotypically mature cells, enriched for BPs involved in the regulation of osteoclastogenesis and immune responses. We also observed perturbed OB dynamic across disease stages with a significant stepwise depletion of pre-OBs WISP2+ and a concomitant expansion of dysfunctional pre-OBs, together with a strong inverse correlation between pre-OBs WISP2+ and tumor burden (r:-0.74; p:0.0041). This pattern was consistent between SMM progressors and non-progressors. In MM, pre-OBs WISP2+ exhibited an upregulated senescence‑associated secretory phenotype and a compromised osteogenic differentiation potential, whereas pre-OBs upregulated osteoblast-inhibitory genes and showed enrichment of glutamine and glutamate metabolic processes. Finally, preliminary interactome analyses (12,129 paired immune and tumor cells) revealed distinct cross-talk signatures within the bone niche.Conclusions: Single-cell profiling highlights profound remodeling of the osteoblastic lineage across precursors disease and MM, marked by depletion of pro-osteogenic pre-OBs WISP2+ and expansion of dysfunctional subsets. This work provides a detailed map of OB dynamics in monoclonal gammopathies showing that transcriptional shifts mirror skeletal niche disruption and may serve as early indicators of tumoral progression toward MM. - High glutamate levels in the bone marrow of multiple myeloma patients promote osteoclast formation: a novel target for osteolytic bone disease
Denise Toscani, Oxana Lungu, Martina Chiu, Chiara Maccari, Vincenzo Raimondi, et al.
Leukemia, 2025
Multiple Myeloma (MM) is a glutamine (Gln)-addicted cancer. Consequently, the MM bone microenvironment (BM) is characterized by lower Gln and higher glutamate (Glu) levels than those in pre-malignant monoclonal gammopathies. Such MM-dependent metabolic perturbation impairs osteoblast differentiation in the bone microenvironment, but its effect on osteoclast (OCL) bone resorption is still unknown. We first show that bone marrow mononuclear cells from MM patients release higher levels of Glu compared to those from patients with monoclonal gammopathy of undetermined significance (MGUS) or smoldering multiple myeloma (SMM). This increased Glu production correlates with elevated bone resorption activity. We then demonstrate that Glu stimulates OCL differentiation via the activation of NF-κB-NFATc1 pathway in low-Glu BM samples from pre-malignant patients but not in high-Glu samples of MM patients. Secondly, the early phase of OCL formation was associated with high Glu intracellular content and induction of the Glu transporter EAAT1. Consistently, the pharmacological inhibition of EAAT1 hinders OCL differentiation by blocking the RANKL-dependent signaling pathway and actin cytoskeleton reorganization. Overall, our data indicate that high Glu levels in MM bone marrow are involved in OCL formation, suggesting that targeting Glu transport may represent a novel approach for the prevention of osteolytic lesions in MM patients. - Mechanistic insights into bone destruction in multiple myeloma: Cellular and molecular perspectives
Oxana Lungu, Denise Toscani, Nicola Giuliani
Journal of Bone Oncology, 2025 - Follow-up of humoral and cellular immune responses after the third SARS-CoV-2 vaccine dose in multiple myeloma patients
Vincenzo Raimondi, Paola Storti, Rosanna Vescovini, Valentina Franceschi, Denise Toscani, et al.
Frontiers in Immunology, 2025
The stability of immune responses to SARS-CoV-2 vaccines, especially concerning the cross-reactive recognition of the Omicron variant, remains incompletely characterized in multiple myeloma (MM) patients. This study evaluated humoral responses in 29 MM patients and cellular responses in a subset of 19 MM patients, specific to Wuhan and Omicron spike proteins, between 16 and 26 weeks following the third vaccine dose. After 26 weeks, we highlighted a significant reduction in the neutralizing antibodies to both spikes and the percentages of IFN-γ+CD107a+ spike-specific CD8+ T cells. On the other hand, patients who underwent an additional stimulation between the two time points, through either a fourth vaccine dose or breakthrough infection, showed a significant increase in neutralizing antibodies and stable levels of cytotoxic CD8+ T cells. Additionally, those with only three doses experienced a higher rate of breakthrough infections during the 32-week follow-up period. These findings underscore the waning of vaccine-induced immunity over time and may help benefit-risk evaluation in vaccination strategies in MM patients. - Identification of PSMB4 and PSMD4 as novel target genes correlated with 1q21 amplification in patients with smoldering myeloma and multiple myeloma
Jessica Burroughs Garcia, Paola Storti, Nicolas Thomas Iannozzi, Valentina Marchica, Luca Agnelli, et al.
Haematologica, 2024
Not available. - The impact of CD56 expression in smoldering myeloma patients on early progression
Laura Notarfranchi, Roberta Segreto, Rosanna Vescovini, Anna Benedetta Dalla Palma, Valentina Marchica, et al.
Hematological Oncology, 2023
Our study identified the absence of CD56 as a possible risk factor for early progression in SMM patients at 36 months. Furthermore, our results showed that the lack of CD56 expression could be a factor for a more aggressive disease regardless of the tumoral burden. For this reason, these patients should have a closer follow-up in the first years after the diagnosis of SMM and the expression of CD56 could potentially be used as a biomarker and could be incorporated in future prognostic or predictive scores. This article is protected by copyright. All rights reserved. - The Metabolic Features of Osteoblasts: Implications for Multiple Myeloma (MM) Bone Disease
Oxana Lungu, Denise Toscani, Jessica Burroughs-Garcia, Nicola Giuliani
International Journal of Molecular Sciences, 2023
The study of osteoblast (OB) metabolism has recently received increased attention due to the considerable amount of energy used during the bone remodeling process. In addition to glucose, the main nutrient for the osteoblast lineages, recent data highlight the importance of amino acid and fatty acid metabolism in providing the fuel necessary for the proper functioning of OBs. Among the amino acids, it has been reported that OBs are largely dependent on glutamine (Gln) for their differentiation and activity. In this review, we describe the main metabolic pathways governing OBs’ fate and functions, both in physiological and pathological malignant conditions. In particular, we focus on multiple myeloma (MM) bone disease, which is characterized by a severe imbalance in OB differentiation due to the presence of malignant plasma cells into the bone microenvironment. Here, we describe the most important metabolic alterations involved in the inhibition of OB formation and activity in MM patients. - Molecular Features of the Mesenchymal and Osteoblastic Cells in Multiple Myeloma
Nicolas Thomas Iannozzi, Valentina Marchica, Denise Toscani, Jessica Burroughs Garcìa, Nicola Giuliani, et al.
International Journal of Molecular Sciences, 2022 - Metabolic features of myeloma cells in the context of bone microenvironment: Implication for the pathophysiology and clinic of myeloma bone disease
Vincenzo Raimondi, Denise Toscani, Valentina Marchica, Jessica Burroughs-Garcia, Paola Storti, et al.
Frontiers in Oncology, 2022 - A personalized molecular approach in multiple myeloma: the possible use of RAF/RAS/MEK/ERK and BCL-2 inhibitors
Vincenzo Raimondi, Nicolas Thomas Iannozzi, Jessica Burroughs-Garcìa, Denise Toscani, Paola Storti, et al.
Exploration of Targeted Anti Tumor Therapy, 2022 - Immune response to SARS-CoV-2 mRNA vaccination and booster dose in patients with multiple myeloma and monoclonal gammopathies: impact of Omicron variant on the humoral response
Paola Storti, Valentina Marchica, Rosanna Vescovini, Valentina Franceschi, Luca Russo, et al.
Oncoimmunology, 2022 - [18F](2S,4R)-4-Fluoroglutamine as a New Positron Emission Tomography Tracer in Myeloma
Silvia Valtorta, Denise Toscani, Martina Chiu, Andrea Sartori, Angela Coliva, et al.
Frontiers in Oncology, 2021 - Role of 1q21 in multiple myeloma: From pathogenesis to possible therapeutic targets
Jessica Burroughs Garcìa, Rosa Alba Eufemiese, Paola Storti, Gabriella Sammarelli, Luisa Craviotto, et al.
Cells, 2021 - The role of proteasome inhibitors in multiple myeloma bone disease and bone metastasis: Effects on osteoblasts and osteocytes
Denise Toscani, Luisa Craviotto, Nicola Giuliani
Applied Sciences Switzerland, 2021 - PD-L1/PD-1 Pattern of Expression Within the Bone Marrow Immune Microenvironment in Smoldering Myeloma and Active Multiple Myeloma Patients
Federica Costa, Rosanna Vescovini, Valentina Marchica, Paola Storti, Laura Notarfranchi, et al.
Frontiers in Immunology, 2021 - Myeloma cells deplete bone marrow glutamine and inhibit osteoblast differentiation limiting asparagine availability
Martina Chiu, Denise Toscani, Valentina Marchica, Giuseppe Taurino, Federica Costa, et al.
Cancers, 2020 - Novel approaches to improve myeloma cell killing by monoclonal antibodies
Paola Storti, Federica Costa, Valentina Marchica, Jessica Burroughs-Garcia, Benedetta dalla Palma, et al.
Journal of Clinical Medicine, 2020 - CD14+CD16+ monocytes are involved in daratumumab-mediated myeloma cells killing and in anti-CD47 therapeutic strategy
Paola Storti, Rosanna Vescovini, Federica Costa, Valentina Marchica, Denise Toscani, et al.
British Journal of Haematology, 2020 - Bovine pestivirus is a new alternative virus for multiple myeloma oncolytic virotherapy
Valentina Marchica, Valentina Franceschi, Rosanna Vescovini, Paola Storti, Emanuela Vicario, et al.
Journal of Hematology and Oncology, 2020 - Novel targets for the treatment of relapsing multiple myeloma
Nicola Giuliani, Fabrizio Accardi, Valentina Marchica, Benedetta Dalla Palma, Paola Storti, et al.
Expert Review of Hematology, 2019 - Bone marrow CX3CL1/Fractalkine is a new player of the pro-angiogenic microenvironment in multiple myeloma patients
Valentina Marchica, Denise Toscani, Anna Corcione, Marina Bolzoni, Paola Storti, et al.
Cancers, 2019 - Multiple myeloma-derived exosomes are enriched of amphiregulin (AREG) and activate the epidermal growth factor pathway in the bone microenvironment leading to osteoclastogenesis
Stefania Raimondo, Laura Saieva, Emanuela Vicario, Marzia Pucci, Denise Toscani, et al.
Journal of Hematology and Oncology, 2019
