Longitudinal and combined assessment of 24(S)-hydroxycholesterol and Neurofilament light chain in the early stages of Huntington's disease Lidia Sarro, Marta Valenza, Alessia Mongelli, Alice Passoni, Anna Castaldo, Monica Favagrossa, Laura Pasetto, Valentina Bonetto, Renzo Bagnati, Marina Grisoli, Anna Nigri, Giulia Birolini, Elena Zerbini, Simone Tomè, Marco Masseroli, Erika Salvi, Laura Colombo, Mario Salmona, Elena Cattaneo, Caterina Mariotti Neurobiology of Disease, 2026 Huntington's Disease (HD) is an autosomal dominant neurodegenerative disorder. Recent clinical research has focused on neurofilament light chain protein (NfL), and cholesterol metabolites as potential fluid biomarkers related to disease progression. Our aim was to explore whether the combined longitudinal assessment of NfL and cholesterol-derived metabolites would improve the characterization of early HD stages. We enrolled 96 HD individuals and 63 healthy controls. Disease stage of HD participants was determined using both the system based on Total Motor Score (TMS), and the new HD-Integrated Staging System (HD-ISS). Concentrations of plasma NfL, and cholesterol precursors and metabolites, were measured at baseline and 2-year follow-up. In premanifest individuals, classified according to TMS, we found significantly reduced 24(S)-hydroxycholesterol (24S-OHC), a metabolite entirely derived from brain cholesterol catabolism, and normal level of NfL. When applying the HD-ISS, 24S-OHC was significantly reduced in HD-ISS-1 (25.6 ± 7.6 ng/ml) and HD-ISS-3 (30.5 ± 15.7) compared with controls (39.5 ± 14.1). NfL was increased in HD-ISS-1 (16.1 ± 6.5 pg/ml), HD-ISS-2 (16.1 ± 6.5) and HD-ISS-3 (27.0 ± 6.9) compared with controls (5.5 ± 1.7 pg/ml). In HD-ISS-0, all biomarker concentrations were similar to those of controls. At follow-up, NfL increased in HD-ISS-1 (+20%; p < 0.005). ROC analyses showed that NfL discriminated HD-ISS-0 and HD-ISS-1 from controls. 24S-OHC distinguished HD-ISS-1 from controls. Plasma NfL and 24S-OHC concentrations provide complementary insights into early biological changes in HD, and holds the potential to characterize the transition phase from the pre-symptomatic to the early symptomatic stage.
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.
Corrigendum to “Chronic cholesterol administration to the brain supports complete and long-lasting cognitive and motor amelioration in Huntington's disease” (Pharmacological Research (2023) 194, (S1043661823001792), (10.1016/j.phrs.2023.106823)) Giulia Birolini, Marta Valenza, Ilaria Ottonelli, Francesca Talpo, Lucia Minoli, Andrea Cappelleri, Mauro Bombaci, Claudio Caccia, Caterina Canevari, Arianna Trucco, Valerio Leoni, Alice Passoni, Monica Favagrossa, Maria Rosaria Nucera, Laura Colombo, Saverio Paltrinieri, Renzo Bagnati, Jason Thomas Duskey, Riccardo Caraffi, Maria Angela Vandelli, Franco Taroni, Mario Salmona, Eugenio Scanziani, Gerardo Biella, Barbara Ruozi, Giovanni Tosi, Elena Cattaneo Pharmacological Research, 2025 “The authors would like to inform readers that, because of a clerical mistakes, the samples labeled as ‘striatum’ in Figure 4A-B, Supplementary Figures S3A-D, S3E-H, S3I-L, S4B-F, S4H-L, S4N-R, and Supplementary Tables 1 and 4 actually correspond to thalamus, while the samples labeled as ‘cortex’ contained a mixture of cortical and striatal tissue. Accordingly, in these figures and in the corresponding text, the label ‘striatum’ should be read as ‘thalamus,’ and the label ‘cortex’ as ‘cortex + striatum’. All behavioral, electrophysiological, and immunohistochemical data reported in the paper were obtained from whole-brain or whole-animal studies and therefore remain unaffected. The authors apologize for any inconvenience this may have caused.”
Xanthohumol destabilizes the structure of amyloid-β (Aβ) oligomers and promotes the formation of high-molecular-weight amorphous aggregates Linda Molteni, Chiara Bruzzone, Diletta Ami, Ada De Luigi, Laura Colombo, Luca Moretti, Antonino Natalello, Alessandro Palmioli, Cristina Airoldi International Journal of Biological Macromolecules, 2025 This study provides a comprehensive structural analysis of prenylflavonoids (PFs) interaction with Aβ1–42 oligomers, revealing their potential to modulate amyloid peptide aggregation pathways. Among the tested PFs, xanthohumol (XN), isoxanthohumol (IXN) and 8-prenylnaringenin (8-PN), XN exhibited the strongest anti-amyloidogenic activity, inhibiting Aβ1–42 aggregation at substoichiometric concentrations and stabilizing the peptide in amorphous aggregates. Its higher conformational flexibility drives the distinct interaction profile and protective effect; in fact, XN forms stable complexes with Aβ1–42 oligomers, preventing their transition to β -sheet-rich fibrils. These findings highlight that XN may offer a neuroprotective strategy by redirecting Aβ1–42 aggregation toward less toxic species, in agreement with its previously described ability to reduce Aβ toxicity. Our data dissect the anti-amyloidogenic mechanism of action of XN at molecular level, providing structural insights for the rational design of a new class of Aβ inhibitors. Indeed, conventional drug discovery approaches are often ineffective against amyloid aggregates, due to their dynamic and atypical receptor structures. Instead, preventive and therapeutic strategies should focus on compounds that form stable supramolecular complexes, capable of modulating the structure and toxicity of the aggregates without relying on specific binding sites. Additionally, due also to its anti-oxidant activity, XN, naturally present in hops, shows promise as nutraceutical for Alzheimer's disease prevention. Dietary supplementation with XN could modulate early molecular events linked to neurodegeneration, providing a safe, sustainable approach to neuroprotection. • A cross-scale characterization of prenylflavonoids-Aβ1–42 interaction is provided. • Xanthohumol (XN), isoxanthohumol and 8-prenylnaringenin from hops are investigated. • XN diverts Aβ1–42 peptide aggregation toward the formation of amorphous aggregates. • The study provides structural insights for the design of Aβ aggregation modulators. • The results underscore the high potential of XN in the nutraceutical field.
The AβA2V paradigm: From molecular insights to therapeutic strategies in Alzheimer's disease and primary tauopathies Luisa Diomede, Andrea Conz, Michele Mosconi, Tatiana Stoilova, Matteo Paloni, Matteo Salvalaglio, Alfredo Cagnotto, Laura Colombo, Marcella Catania, Giuseppe Di Fede, Fabrizio Tagliavini, Mario Salmona Pharmacological Research, 2025 (D), which shows significant neuroprotective activity through inhibition of the assembly of amyloid β into amyloid fibrils. The therapeutic potential of this peptide emerges from its ability to reduce amyloid β-induced toxicity, with promising results from studies in human neuroblastoma cells and transgenic animal models.
Thermal effects and biological response of breast and pancreatic cancer cells undergoing gold nanorod-assisted photothermal therapy Leonardo Bianchi, Sara Baroni, Gabriela Paroni, Martina Bruna Violatto, Giulia Yuri Moscatiello, Nicolò Panini, Luca Russo, Fabio Fiordaliso, Laura Colombo, Luisa Diomede, Paola Saccomandi, Paolo Bigini Journal of Photochemistry and Photobiology B Biology, 2024 To increase the therapeutic efficacy of nanoparticle (NP)-assisted photothermal therapy (PTT) and allow for a transition toward the clinical setting, it is pivotal to characterize the thermal effect induced in cancer cells and correlate it with the cell biological response, namely cell viability and cell death pathways. This study quantitatively evaluated the effects of gold nanorod (GNR)-assisted near-infrared (NIR) PTT on two different cancer cell lines, the 4T1 triple-negative breast cancer cells and the Pan02 pancreatic cancer cells. The interaction between nanomaterials and biological matrices was investigated in terms of GNR internalization and effect on cell viability at different GNR concentrations. GNR-mediated PTT was executed on both cell lines, at the same treatment settings to allow a straightforward comparison, and real-time monitored through thermographic imaging. A thermal analysis based on various parameters ( i.e. , maximum absolute temperature, maximum temperature change, temperature variation profile, area under the time-temperature change curve, effective thermal enhancement (ETE), and time constants) was performed to evaluate the treatment thermal outcome. While GNR treatment and NIR laser irradiation alone did not cause cell toxicity in the selected settings, their combination induced a significant reduction of cell viability in both cell lines. At the optimal experimental condition ( i.e. , 6 μg/mL of GNRs and 4.5 W/cm 2 laser power density), GNR-assisted PTT reduced the cell viability of 4T1 and Pan02 cells by 94% and 87% and it was associated with maximum temperature changes of 25 °C and 29 °C ( i.e. , ∼1.8-fold increase compared to the laser-only condition), maximum absolute temperatures of 55 °C and 54 °C, and ETE values of 78% and 81%, for 4T1 and Pan02 cells, correspondingly. Also, the increase in the GNR concentration led to a decrease in the time constants, denoting faster heating kinetics upon irradiation. Furthermore, the thermal analysis parameters were correlated with the extent of cell death. Twelve hours after NIR exposure, GNR-assisted PTT was found to mainly trigger secondary apoptosis in both cell lines. The proposed study provides relevant insights into the relationship between temperature history and biological responses in the context of PTT. The findings contribute to the development of a universal methodology for evaluating thermal sensitivity upon NP-assisted PTT on different cell types and lay the groundwork for future translational studies. • GNR-assisted PTT significantly reduced breast and pancreatic cancer cell viability. • The thermal effects of GNR-mediated PTT were correlated with cell biological response. • Increasing GNR concentration led to faster heating kinetics upon irradiation. • 12 h post-exposure, GNR-mediated PTT was found to mainly trigger secondary apoptosis.
A cutting-edge approach based on UHPLC-MS to simultaneously investigate oxysterols and cholesterol precursors in biological samples: Validation in Huntington's disease mouse model Alice Passoni, Monica Favagrossa, Marta Valenza, Giulia Birolini, Alessia Lanno, Caterina Mariotti, Elena Cattaneo, Mario Salmona, Laura Colombo, Renzo Bagnati Talanta Open, 2024 Brain is most cholesterol-rich organ in the body. Since cholesterol does not cross the blood brain barrier, its metabolism is provided in situ by astrocytes and neurons, and it is crucial for maintaining sterol levels and neuronal integrity and function. Recent studies have shown that the levels of cholesterol precursors and metabolites are lower in the brains of animal models of Huntington's disease (HD) while reduced levels of its catabolite are detected in the plasma of patients. In this study, we introduce a novel analytical method designed to fulfil the complex analytical requirements associated with cholesterol metabolites detection in neurodegenerative disorders. The method allows for the simultaneous quantification of a specific set of oxysterols along with cholesterol precursors in biological samples. The proposed method uses an Ultra-High-Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) system operating in multiple reaction monitoring (MRM). Since sterols can be found in biological matrices in either free form or esterified to various fatty acids, a three-step extraction procedure was devised, consisting of alkaline hydrolysis, liquid-liquid extraction and final concentration omitting the need for a solid-phase extraction (SPE) step. The validated method achieved a detection limit of 10 ng/mL in plasma and 1 ng/mg in brain tissue, reaching a comparable sensitivity to previously published LC-MS and GC-MS methods. All target analytes were separated on a reverse-phase column employing a segmented gradient and a temperature ramp. This strategy enabled the elution and separation of all selected metabolites within a 30-minutes timeframe. This innovative approach was employed to quantify cholesterol metabolites in both plasma and brain samples from wild-type (WT) and R6/2 mice, a mouse model of HD. The results obtained from the sample analysis highlighted a significant reduction in desmosterol levels in the R6/2 brain at 12 weeks. In conclusion, the proposed method paves the way for further development of high-sensitive and reproducible protocols to comprehensively investigate simultaneous alterations in both cholesterol biosynthesis and catabolism in HD samples.
Genetic deletion of astrocytic calcineurin B1 prevents cognitive impairment and neuropathology development in acute and chronic mouse models of Alzheimer's disease Laura Tapella, Giulia Dematteis, Pietro La Vitola, Susanna Leva, Elisa Tonelli, Marco Raddi, Marta Delconti, Letizia Dacomo, Alberto La Macchia, Elisa Murari, Maria Talmon, Justyna Malecka, Gabriela Chrostek, Mariagrazia Grilli, Laura Colombo, Mario Salmona, Gianluigi Forloni, Armando A. Genazzani, Claudia Balducci, Dmitry Lim GLIA, 2024 Alzheimer's disease (AD) represents an urgent yet unmet challenge for modern society, calling for exploration of innovative targets and therapeutic approaches. Astrocytes, main homeostatic cells in the CNS, represent promising cell‐target. Our aim was to investigate if deletion of the regulatory CaNB1 subunit of calcineurin in astrocytes could mitigate AD‐related memory deficits, neuropathology, and neuroinflammation. We have generated two, acute and chronic, AD mouse models with astrocytic CaNB1 ablation (ACN‐KO). In the former, we evaluated the ability of β‐amyloid oligomers (AβOs) to impair memory and activate glial cells once injected in the cerebral ventricle of conditional ACN‐KO mice. Next, we generated a tamoxifen‐inducible astrocyte‐specific CaNB1 knock‐out in 3xTg‐AD mice (indACNKO‐AD). CaNB1 was deleted, by tamoxifen injection, in 11.7‐month‐old 3xTg‐AD mice for 4.4 months. Spatial memory was evaluated using the Barnes maze; β‐amyloid plaques burden, neurofibrillary tangle deposition, reactive gliosis, and neuroinflammation were also assessed. The acute model showed that ICV injected AβOs in 2‐month‐old wild type mice impaired recognition memory and fostered a pro‐inflammatory microglia phenotype, whereas in ACN‐KO mice, AβOs were inactive. In indACNKO‐AD mice, 4.4 months after CaNB1 depletion, we found preservation of spatial memory and cognitive flexibility, abolishment of amyloidosis, and reduction of neurofibrillary tangles, gliosis, and neuroinflammation. Our results suggest that ACN is crucial for the development of cognitive impairment, AD neuropathology, and neuroinflammation. Astrocyte‐specific CaNB1 deletion is beneficial for both the abolishment of AβO‐mediated detrimental effects and treatment of ongoing AD‐related pathology, hence representing an intriguing target for AD therapy.
Cell-Free and In Vivo Characterization of the Inhibitory Activity of Lavado Cocoa Flavanols on the Amyloid Protein Ataxin-3: Toward New Approaches against Spinocerebellar Ataxia Type 3 Barbara Sciandrone, Alessandro Palmioli, Carlotta Ciaramelli, Roberta Pensotti, Laura Colombo, Maria Elena Regonesi, Cristina Airoldi ACS Chemical Neuroscience, 2024 Spinocerebellar ataxia type 3 (SCA3) is a neurodegenerative disorder characterized by ataxia and other neurological manifestations, with a poor prognosis and a lack of effective therapies. The amyloid aggregation of the ataxin-3 protein is a hallmark of SCA3 and one of the main biochemical events prompting its onset, making it a prominent target for the development of preventive and therapeutic interventions. Here, we tested the efficacy of an aqueous Lavado cocoa extract and its polyphenolic components against ataxin-3 aggregation and neurotoxicity. The combination of biochemical assays and atomic force microscopy morphological analysis provided clear evidence of cocoa flavanols' ability to hinder ATX3 amyloid aggregation through direct physical interaction, as assessed by NMR spectroscopy. The chemical identity of the flavanols was investigated by ultraperformance liquid chromatography-high-resolution mass spectrometry. The use of the preclinical model Caenorhabditis elegans allowed us to demonstrate cocoa flavanols' ability to ameliorate ataxic phenotypes in vivo. To the best of our knowledge, Lavado cocoa is the first natural source whose extract is able to directly interfere with ATX3 aggregation, leading to the formation of off-pathway species.
Chronic cholesterol administration to the brain supports complete and long-lasting cognitive and motor amelioration in Huntington's disease Giulia Birolini, Marta Valenza, Ilaria Ottonelli, Francesca Talpo, Lucia Minoli, Andrea Cappelleri, Mauro Bombaci, Claudio Caccia, Caterina Canevari, Arianna Trucco, Valerio Leoni, Alice Passoni, Monica Favagrossa, Maria Rosaria Nucera, Laura Colombo, Saverio Paltrinieri, Renzo Bagnati, Jason Thomas Duskey, Riccardo Caraffi, Maria Angela Vandelli, Franco Taroni, Mario Salmona, Eugenio Scanziani, Gerardo Biella, Barbara Ruozi, Giovanni Tosi, Elena Cattaneo Pharmacological Research, 2023
Micro- and Nanoplastics’ Effects on Protein Folding and Amyloidosis Joseph Windheim, Laura Colombo, Nora C. Battajni, Luca Russo, Alfredo Cagnotto, Luisa Diomede, Paolo Bigini, Elena Vismara, Ferdinando Fiumara, Silvia Gabbrielli, Alfonso Gautieri, Gemma Mazzuoli-Weber, Mario Salmona, Luca Colnaghi International Journal of Molecular Sciences, 2022
C. elegans detects toxicity of traumatic brain injury generated tau Elisa R. Zanier, Maria Monica Barzago, Gloria Vegliante, Margherita Romeo, Elena Restelli, Ilaria Bertani, Carmina Natale, Luca Colnaghi, Laura Colombo, Luca Russo, Edoardo Micotti, Luana Fioriti, Roberto Chiesa, Luisa Diomede Neurobiology of Disease, 2021
Monomeric Aβ1-42 and RAGE: Key players in neuronal differentiation Sabrina Piras, Anna L. Furfaro, Alessandra Piccini, Mario Passalacqua, Roberta Borghi, Enrico Carminati, Alessia Parodi, Laura Colombo, Mario Salmona, Maria A. Pronzato, Umberto M. Marinari, Massimo Tabaton, Mariapaola Nitti Neurobiology of Aging, 2014
Good gene, bad gene: New APP variant may be both Giuseppe Di Fede, Marcella Catania, Michela Morbin, Giorgio Giaccone, Maria Luisa Moro, Roberta Ghidoni, Laura Colombo, Massimo Messa, Alfredo Cagnotto, Margherita Romeo, Matteo Stravalaci, Luisa Diomede, Marco Gobbi, Mario Salmona, Fabrizio Tagliavini Progress in Neurobiology, 2012
β-Amyloid 1-42 induces physiological transcriptional regulation of BACE1 Alessandra Piccini, Roberta Borghi, Michela Guglielmotto, Elena Tamagno, Gabriella Cirmena, Anna Garuti, Valeria Pollero, Sergio Cammarata, Michele Fornaro, Massimo Messa, Laura Colombo, Mario Salmona, George Perry, Massimo Tabaton Journal of Neurochemistry, 2012
Curcumin derivatives as new ligands of Aβ peptides Cristina Airoldi, Cristiano Zona, Erika Sironi, Laura Colombo, Massimo Messa, Dario Aurilia, Maria Gregori, Massimo Masserini, Mario Salmona, Francesco Nicotra, Barbara La Ferla Journal of Biotechnology, 2011
Synthetic amyloid-β oligomers impair long-term memory independently of cellular prion protein Claudia Balducci, Marten Beeg, Matteo Stravalaci, Antonio Bastone, Alessandra Sclip, Emiliano Biasini, Laura Tapella, Laura Colombo, Claudia Manzoni, Tiziana Borsello, Roberto Chiesa, Marco Gobbi, Mario Salmona, Gianluigi Forloni Proceedings of the National Academy of Sciences of the United States of America, 2010
A recessive mutation in the APP gene with dominant-negative effect on amyloidogenesis Giuseppe Di Fede, Marcella Catania, Michela Morbin, Giacomina Rossi, Silvia Suardi, Giulia Mazzoleni, Marco Merlin, Anna Rita Giovagnoli, Sara Prioni, Alessandra Erbetta, Chiara Falcone, Marco Gobbi, Laura Colombo, Antonio Bastone, Marten Beeg, Claudia Manzoni, Bruna Francescucci, Alberto Spagnoli, Laura Cantù, Elena Del Favero, Efrat Levy, Mario Salmona, Fabrizio Tagliavini Science, 2009
Role of plasminogen in propagation of scrapie Mario Salmona, Raffaella Capobianco, Laura Colombo, Ada De Luigi, Giacomina Rossi, Michela Mangieri, Giorgio Giaccone, Elena Quaglio, Roberto Chiesa, Maria Benedetta Donati, Fabrizio Tagliavini, Gianluigi Forloni Journal of Virology, 2005
Structural Properties of Gerstmann-Sträussler-Scheinker Disease Amyloid Protein Mario Salmona, Michela Morbin, Tania Massignan, Laura Colombo, Giulia Mazzoleni, Raffaella Capobianco, Luisa Diomede, Florian Thaler, Luca Mollica, Giovanna Musco, Joseph J. Kourie, Orso Bugiani, Deepak Sharma, Hideyo Inouye, Daniel A. Kirschner, Gianluigi Forloni, Fabrizio Tagliavini Journal of Biological Chemistry, 2003
Evaluation of quinacrine treatment for prion diseases A. Barret, F. Tagliavini, G. Forloni, C. Bate, M. Salmona, L. Colombo, A. De Luigi, L. Limido, S. Suardi, G. Rossi, F. Auvré, K. T. Adjou, N. Salès, A. Williams, C. Lasmézas, J. P. Deslys Journal of Virology, 2003
Synthetic miniprion PrP106 Valentina Bonetto, Tania Massignan, Roberto Chiesa, Michela Morbin, Giulia Mazzoleni, Luisa Diomede, Nadia Angeretti, Laura Colombo, Gianluigi Forloni, Fabrizio Tagliavini, Mario Salmona Journal of Biological Chemistry, 2002
Tetracyclines affect prion infectivity Gianluigi Forloni, Selina Iussich, Tazeen Awan, Laura Colombo, Nadia Angeretti, Laura Girola, Ilaria Bertani, Giorgio Poli, Maria Caramelli, Maria Grazia Bruzzone, Laura Farina, Lucia Limido, Giacomina Rossi, Giorgio Giaccone, James W. Ironside, Orso Bugiani, Mario Salmona, Fabrizio Tagliavini Proceedings of the National Academy of Sciences of the United States of America, 2002