2007-12-01 to 2010-12-31 | PhD in Biochemistry
2000-10-01 to 2006-03-31 | Master's Degree in Medical Biotechnologies (Department of Histology, Microbiology and Medical Biotechnologies)
Tau oligomers impair memory and synaptic plasticity through the cellular prion protein C. Balducci, Franca Orsini, M. Cerović, M. Beeg, Beatrice Rocutto, L. Dacomo, Antonio Masone, Eleonora Busani, Ilaria Raimondi, Giada Lavigna, Po-Tao Chen, Susanna Leva, L. Colombo, C. Zucchelli, Giovanna Musco, N. Kanaan, Marco Gobbi, Roberto Chiesa, Luana Fioriti, G. Forloni Acta Neuropathologica Communications, 2025 Deposition of abnormally phosphorylated tau aggregates is a central event leading to neuronal dysfunction and death in Alzheimer’s disease (AD) and other tauopathies. Among tau aggregates, oligomers (TauOs) are considered the most toxic. AD brains show significant increase in TauOs compared to healthy controls, their concentration correlating with the severity of cognitive deficits and disease progression. In vitro and in vivo neuronal TauO exposure leads to synaptic and cognitive dysfunction, but their mechanisms of action are unclear. Evidence suggests that the cellular prion protein (PrPC) may act as a mediator of TauO neurotoxicity, as previously proposed for β-amyloid and α-synuclein oligomers. To investigate whether PrPC mediates TauO detrimental activities, we compared their effects on memory and synaptic plasticity in wild type (WT) and PrPC knockout (Prnp0/0) mice. Intracerebroventricular injection of TauOs significantly impaired recognition memory in WT but not in Prnp0/0 mice. Similarly, TauOs inhibited long-term potentiation in acute hippocampal slices from WT but not Prnp0/0 mice. Surface plasmon resonance indicated a high-affinity binding between TauOs and PrPC with a KD of 20–50 nM. Immunofluorescence analysis of naïve and PrPC-overexpressing HEK293 cells exposed to TauOs showed a PrPC dose-dependent association of TauOs with cells over time, and their co-localization with PrPC on the plasma membrane and in intracellular compartments, suggesting PrPC-may play a role in TauO internalization. These findings support the concept that PrPC mediates the detrimental activities of TauOs through a direct interaction, suggesting that targeting this interaction might be a promising therapeutic strategy for AD and other tauopathies.
Therapeutic targeting of cellular prion protein: toward the development of dual mechanism anti-prion compounds Antonio Masone, Chiara Zucchelli, Enrico Caruso, Giovanna Musco, Roberto Chiesa Neural Regeneration Research, 2025 PrPSc, a misfolded, aggregation-prone isoform of the cellular prion protein (PrPC), is the infectious prion agent responsible for fatal neurodegenerative diseases of humans and other mammals. PrPSc can adopt different pathogenic conformations (prion strains), which can be resistant to potential drugs, or acquire drug resistance, posing challenges for the development of effective therapies. Since PrPC is the obligate precursor of any prion strain and serves as the mediator of prion neurotoxicity, it represents an attractive therapeutic target for prion diseases. In this minireview, we briefly outline the approaches to target PrPC and discuss our recent identification of Zn(II)-BnPyP, a PrPC-targeting porphyrin with an unprecedented bimodal mechanism of action. We argue that in-depth understanding of the molecular mechanism by which Zn(II)-BnPyP targets PrPC may lead toward the development of a new class of dual mechanism anti-prion compounds.
SP140 represses specific loci by recruiting polycomb repressive complex 2 and NuRD complex Simone Tamburri, Chiara Zucchelli, Vittoria Matafora, Ettore Zapparoli, Zivojin Jevtic, Francesco Farris, Fabio Iannelli, Giovanna Musco, Angela Bachi Nucleic Acids Research, 2025 SP140, a lymphocytic-restricted protein, is an epigenetic reader working as a corepressor of genes implicated in inflammation and orchestrating macrophage transcriptional programs to maintain cellular identity. Reduced SP140 expression is associated both to autoimmune diseases and blood cancers. However, the molecular mechanisms that link SP140 altered protein levels to detrimental effects on the immune response and cellular growth, as well as the interactors through which SP140 promotes gene silencing, remain elusive. In this work, we have applied a multi-omics approach (i.e. interactomics, ChIP-seq and proteomics) in two Burkitt lymphoma cell lines to identify both interactors and target genes of endogenous SP140. We found that SP140 interacts with the PRC2 and NuRD complexes, and we showed that these interactions are functional as SP140 directs H3K27me3 deposition and NuRD binding on a set of target genes implicated in cellular growth and leukemia progression.
The C-terminal PHDVC5HCH tandem domain of NSD2 is a combinatorial reader of unmodified H3K4 and tri-methylated H3K27 that regulates transcription of cell adhesion genes in multiple myeloma A. Berardi, C. Kaestner, Michela Ghitti, G. Quilici, Paolo Cocomazzi, Jianping Li, Federico Ballabio, C. Zucchelli, Stefan Knapp, Jonathan D. Licht, Giovanna Musco Nucleic Acids Research, 2025 Histone methyltransferase NSD2 (MMSET) overexpression in multiple myeloma (MM) patients plays an important role in the development of this disease subtype. Through the expansion of transcriptional activating H3K36me2 and the suppression of repressive H3K27me3 marks, NSD2 activates an aberrant set of genes that contribute to myeloma growth, adhesive and invasive activities. NSD2 transcriptional activity also depends on its non-catalytic domains, which facilitate its recruitment to chromatin through histone binding. In this study, using NMR, ITC and molecular dynamics simulations, we show that the tandem PHD domain of NSD2 (PHDVC5HCHNSD2) is a combinatorial reader of unmodified histone H3K4 and tri-methylated H3K27 (H3K27me3). This is the first PHD tandem cassette known to decode the methylation status of H3K27. Importantly, in a NSD2-dependent MM cellular model, we show that expression of NSD2 mutants, engineered to disrupt the interaction between H3K27me3 and PHDVC5HCH, display in comparison to wild-type NSD2: incomplete loss of H3K27 methylation throughout the genome, decreased activation of adhesive properties and cell adhesion genes, and a decrease of the corresponding H3K27ac signal at promoters. Collectively, these data suggest that the PHDVC5HCH domain of NSD2 plays an important role in modulating gene expression and chromatin modification, providing new opportunities for pharmacological intervention.
The acidic intrinsically disordered region of the inflammatory mediator HMGB1 mediates fuzzy interactions with CXCL12 Malisa Vittoria Mantonico, Federica De Leo, Giacomo Quilici, Liam Sean Colley, Francesco De Marchis, Massimo Crippa, Rosanna Mezzapelle, Tim Schulte, Chiara Zucchelli, Chiara Pastorello, Camilla Carmeno, Francesca Caprioglio, Stefano Ricagno, Gabriele Giachin, Michela Ghitti, Marco Emilio Bianchi, Giovanna Musco Nature Communications, 2024 Chemokine heterodimers activate or dampen their cognate receptors during inflammation. The CXCL12 chemokine forms with the fully reduced (fr) alarmin HMGB1 a physiologically relevant heterocomplex (frHMGB1•CXCL12) that synergically promotes the inflammatory response elicited by the G-protein coupled receptor CXCR4. The molecular details of complex formation were still elusive. Here we show by an integrated structural approach that frHMGB1•CXCL12 is a fuzzy heterocomplex. Unlike previous assumptions, frHMGB1 and CXCL12 form a dynamic equimolar assembly, with structured and unstructured frHMGB1 regions recognizing the CXCL12 dimerization surface. We uncover an unexpected role of the acidic intrinsically disordered region (IDR) of HMGB1 in heterocomplex formation and its binding to CXCR4 on the cell surface. Our work shows that the interaction of frHMGB1 with CXCL12 diverges from the classical rigid heterophilic chemokines dimerization. Simultaneous interference with multiple interactions within frHMGB1•CXCL12 might offer pharmacological strategies against inflammatory conditions.
Erratum: A tetracationic porphyrin with dual anti-prion activity (iScience (2023) 26(9), (S2589004223015572), (10.1016/j.isci.2023.107480)) Antonio Masone, Chiara Zucchelli, Enrico Caruso, Giada Lavigna, Hasier Eraña, Gabriele Giachin, Laura Tapella, Liliana Comerio, Elena Restelli, Ilaria Raimondi, Saioa R. Elezgarai, Federica De Leo, Giacomo Quilici, Lorenzo Taiarol, Marvin Oldrati, Nuria L. Lorenzo, Sandra García-Martínez, Alfredo Cagnotto, Jacopo Lucchetti, Marco Gobbi, Ilaria Vanni, Romolo Nonno, Michele A. Di Bari, Mark D. Tully, Valentina Cecatiello, Giuseppe Ciossani, Sebastiano Pasqualato, Eelco Van Anken, Mario Salmona, Joaquín Castilla, Jesús R. Requena, Stefano Banfi, Giovanna Musco, Roberto Chiesa Iscience, 2023 (iScience 26, 107480; September 15, 2023) In the originally published version of this article, an error occurred during figure preparation, where the authors duplicated the lower middle panel in Figure 1G. This error has been corrected online with the insertion of the correct image in the lower right panel. The authors apologize for this error and any confusion it may have caused.Figure 1. Zn(II)-BnPyP down-regulates PrPC (corrected)View Large Image Figure ViewerDownload Hi-res image Download (PPT) A tetracationic porphyrin with dual anti-prion activityMasone et al.iScienceJuly 27, 2023In BriefPharmacology; Molecular neuroscience; Cell biology Full-Text PDF Open Access
A tetracationic porphyrin with dual anti-prion activity Antonio Masone, Chiara Zucchelli, Enrico Caruso, Giada Lavigna, Hasier Eraña, Gabriele Giachin, Laura Tapella, Liliana Comerio, Elena Restelli, Ilaria Raimondi, Saioa R. Elezgarai, Federica De Leo, Giacomo Quilici, Lorenzo Taiarol, Marvin Oldrati, Nuria L. Lorenzo, Sandra García-Martínez, Alfredo Cagnotto, Jacopo Lucchetti, Marco Gobbi, Ilaria Vanni, Romolo Nonno, Michele A. Di Bari, Mark D. Tully, Valentina Cecatiello, Giuseppe Ciossani, Sebastiano Pasqualato, Eelco Van Anken, Mario Salmona, Joaquín Castilla, Jesús R. Requena, Stefano Banfi, Giovanna Musco, Roberto Chiesa Iscience, 2023 Prions are deadly infectious agents made of PrP Sc , a misfolded variant of the cellular prion protein (PrP C ) which self-propagates by inducing misfolding of native PrP C . PrP Sc can adopt different pathogenic conformations (prion strains), which can be resistant to potential drugs, or acquire drug resistance, hampering the development of effective therapies. We identified Zn(II)-BnPyP, a tetracationic porphyrin that binds to distinct domains of native PrP C , eliciting a dual anti-prion effect. Zn(II)-BnPyP binding to a C-terminal pocket destabilizes the native PrP C fold, hindering conversion to PrP Sc ; Zn(II)-BnPyP binding to the flexible N-terminal tail disrupts N- to C-terminal interactions, triggering PrP C endocytosis and lysosomal degradation, thus reducing the substrate for PrP Sc generation. Zn(II)-BnPyP inhibits propagation of different prion strains in vitro , in neuronal cells and organotypic brain cultures. These results identify a PrP C -targeting compound with an unprecedented dual mechanism of action which might be exploited to achieve anti-prion effects without engendering drug resistance.
Mechanism of action of the tumor vessel targeting agent NGR-hTNF: Role of both NGR peptide and hTNF in cell binding and signaling Barbara Valentinis, Simona Porcellini, Claudia Asperti, Manuela Cota, Dan Zhou, Paola Di Matteo, Gianpiero Garau, Chiara Zucchelli, Nilla Roberta Avanzi, Gian Paolo Rizzardi, Massimo Degano, Giovanna Musco, Catia Traversari International Journal of Molecular Sciences, 2019 NGR-hTNF is a therapeutic agent for a solid tumor that specifically targets angiogenic tumor blood vessels, through the NGR motif. Its activity has been assessed in several clinical studies encompassing tumors of different histological types. The drug’s activity is based on an improved permeabilization of newly formed tumor vasculature, which favors intratumor penetration of chemotherapeutic agents and leukocyte trafficking. This work investigated the binding and the signaling properties of the NGR-hTNF, to elucidate its mechanism of action. The crystal structure of NGR-hTNF and modeling of its interaction with TNFR suggested that the NGR region is available for binding to a specific receptor. Using 2D TR-NOESY experiments, this study confirmed that the NGR-peptides binds to a specific CD13 isoform, whose expression is restricted to tumor vasculature cells, and to some tumor cell lines. The interaction between hTNF or NGR-hTNF with immobilized TNFRs showed similar kinetic parameters, whereas the competition experiments performed on the cells expressing both TNFR and CD13 showed that NGR-hTNF had a higher binding affinity than hTNF. The analysis of the NGR-hTNF-triggered signal transduction events showed a specific impairment in the activation of pro-survival pathways (Ras, Erk and Akt), compared to hTNF. Since a signaling pattern identical to NGR-hTNF was obtained with hTNF and NGR-sequence given as distinct molecules, the inhibition observed on the survival pathways was presumably due to a direct effect of the NGR-CD13 engagement on the TNFR signaling pathway. The reduced activation of the pro survival pathways induced by NGR-hTNF correlated with the increased caspases activation and reduced cell survival. This study demonstrates that the binding of the NGR-motif to CD13 determines not only the homing of NGR-hTNF to tumor vessels, but also the increase in its antiangiogenic activity.
Sp140 is a multi-SUMO-1 target and its PHD finger promotes SUMOylation of the adjacent Bromodomain Chiara Zucchelli, Simone Tamburri, Giuseppe Filosa, Michela Ghitti, Giacomo Quilici, Angela Bachi, Giovanna Musco Biochimica Et Biophysica Acta General Subjects, 2019 Human Sp140 protein is a leukocyte-specific member of the speckled protein (Sp) family (Sp100, Sp110, Sp140, Sp140L), a class of multi-domain nuclear proteins involved in intrinsic immunity and transcriptional regulation. Sp140 regulates macrophage transcriptional program and is implicated in several haematologic malignancies. Little is known about Sp140 structural domains and its post-translational modifications. We used mass spectrometry and biochemical experiments to investigate endogenous Sp140 SUMOylation in Burkitt's Lymphoma cells and Sp140 SUMOylation sites in HEK293T cells, FLAG-Sp140 transfected and His6-SUMO-1T95K infected. NMR spectroscopy and in vitro SUMOylation reactions were applied to investigate the role of Sp140 PHD finger in the SUMOylation of the adjacent BRD. Endogenous Sp140 is a SUMO-1 target, whereby FLAG-Sp140 harbors at least 13 SUMOylation sites distributed along the protein sequence, including the BRD. NMR experiments prove direct binding of the SUMO E2 ligase Ubc9 and SUMO-1 to PHD-BRDSp140. In vitro SUMOylation reactions show that the PHDSp140 behaves as SUMO E3 ligase, assisting intramolecular SUMOylation of the adjacent BRD. Sp140 is multi-SUMOylated and its PHD finger works as versatile protein-protein interaction platform promoting intramolecular SUMOylation of the adjacent BRD. Thus, combinatorial association of Sp140 chromatin binding domains generates a multifaceted interaction scaffold, whose function goes beyond the canonical histone recognition. The addition of Sp140 to the increasing lists of multi-SUMOylated proteins opens new perspectives for molecular studies on Sp140 transcriptional activity, where SUMOylation could represent a regulatory route and a docking surface for the recruitment and assembly of leukocyte-specific transcription regulators.
