{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T01:35:22Z","timestamp":1790904922981,"version":"4.1.0"},"reference-count":34,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2024,1,17]],"date-time":"2024-01-17T00:00:00Z","timestamp":1705449600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Regione Toscana (Bando Salute 2018)","award":["PE0000006"],"award-info":[{"award-number":["PE0000006"]}]},{"name":"University of Florence","award":["PE0000006"],"award-info":[{"award-number":["PE0000006"]}]},{"name":"#NEXTGENERATIONEU (NGEU)","award":["PE0000006"],"award-info":[{"award-number":["PE0000006"]}]},{"name":"FONDAZIONE Cassa di Risparmio Pistoia e Pescia, Bando Giovani@RicercaScientifica 2021","award":["PE0000006"],"award-info":[{"award-number":["PE0000006"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Gaucher disease (GD) is a rare genetic metabolic disorder characterized by a dysfunction of the lysosomal glycoside hydrolase glucocerebrosidase (GCase) due to mutations in the gene GBA1, leading to the cellular accumulation of glucosylceramide (GlcCer). While most of the current research focuses on the primary accumulated material, lesser attention has been paid to secondary storage materials and their reciprocal intertwining. By using a novel approach based on flow cytometry and fluorescent labelling, we monitored changes in storage materials directly in fibroblasts derived from GD patients carrying N370S\/RecNcil and homozygous L444P or R131C mutations with respect to wild type. In L444P and R131C fibroblasts, we detected not only the primary accumulation of GlcCer accumulation but also a considerable secondary increase in GM1 storage, comparable with the one observed in infantile patients affected by GM1 gangliosidosis. In addition, the ability of a trivalent trihydroxypiperidine iminosugar compound (CV82), which previously showed good pharmacological chaperone activity on GCase enzyme, to reduce the levels of storage materials in L444P and R131C fibroblasts was tested. Interestingly, treatment with different concentrations of CV82 led to a significant reduction in GM1 accumulation only in L444P fibroblasts, without significantly affecting GlcCer levels. The compound CV82 was selective against the GCase enzyme with respect to the \u03b2-Galactosidase enzyme, which was responsible for the catabolism of GM1 ganglioside. The reduction in GM1-ganglioside level cannot be therefore ascribed to a direct action of CV82 on \u03b2-Galactosidase enzyme, suggesting that GM1 decrease is rather related to other unknown mechanisms that follow the direct action of CV82 on GCase. In conclusion, this work indicates that the tracking of secondary storages can represent a key step for a better understanding of the pathways involved in the severity of GD, also underlying the importance of developing drugs able to reduce both primary and secondary storage-material accumulations in GD.<\/jats:p>","DOI":"10.3390\/molecules29020453","type":"journal-article","created":{"date-parts":[[2024,1,17]],"date-time":"2024-01-17T07:41:28Z","timestamp":1705477288000},"page":"453","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Identification of GM1-Ganglioside Secondary Accumulation in Fibroblasts from Neuropathic Gaucher Patients and Effect of a Trivalent Trihydroxypiperidine Iminosugar Compound on Its Storage Reduction"],"prefix":"10.3390","volume":"29","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6832-1703","authenticated-orcid":false,"given":"Costanza","family":"Ceni","sequence":"first","affiliation":[{"name":"Department of Chemistry \u201cU. Schiff\u201d (DICUS), University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy"},{"name":"European Laboratory for Non-Linear Spectroscopy (LENS), University of Florence, 50019 Sesto Fiorentino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9899-0836","authenticated-orcid":false,"given":"Francesca","family":"Clemente","sequence":"additional","affiliation":[{"name":"Department of Chemistry \u201cU. Schiff\u201d (DICUS), University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5344-2310","authenticated-orcid":false,"given":"Francesca","family":"Mangiavacchi","sequence":"additional","affiliation":[{"name":"Department of Chemistry \u201cU. Schiff\u201d (DICUS), University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8336-383X","authenticated-orcid":false,"given":"Camilla","family":"Matassini","sequence":"additional","affiliation":[{"name":"Department of Chemistry \u201cU. Schiff\u201d (DICUS), University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5170-6757","authenticated-orcid":false,"given":"Rodolfo","family":"Tonin","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children\u2019s Hospital IRCCS, 50139 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1413-2624","authenticated-orcid":false,"given":"Anna","family":"Caciotti","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children\u2019s Hospital IRCCS, 50139 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1520-188X","authenticated-orcid":false,"given":"Federica","family":"Feo","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children\u2019s Hospital IRCCS, 50139 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8440-865X","authenticated-orcid":false,"given":"Domenico","family":"Coviello","sequence":"additional","affiliation":[{"name":"Laboratory of Human Genetics, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2890-8179","authenticated-orcid":false,"given":"Amelia","family":"Morrone","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children\u2019s Hospital IRCCS, 50139 Florence, Italy"},{"name":"Department of Neurosciences, Psychology, Drug Research and Child Health (NEUROFARBA), University of Florence, 50121 Florence, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6766-4624","authenticated-orcid":false,"given":"Francesca","family":"Cardona","sequence":"additional","affiliation":[{"name":"Department of Chemistry \u201cU. Schiff\u201d (DICUS), University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4031-7235","authenticated-orcid":false,"given":"Martino","family":"Calamai","sequence":"additional","affiliation":[{"name":"European Laboratory for Non-Linear Spectroscopy (LENS), University of Florence, 50019 Sesto Fiorentino, Italy"},{"name":"National Institute of Optics-National Research Council (CNR-INO), 50019 Sesto Fiorentino, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Castillon, G., Chang, S.-C., and Moride, Y. (2022). Global Incidence and Prevalence of Gaucher Disease: A Targeted Literature Review. J. Clin. Med., 12.","DOI":"10.3390\/jcm12010085"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1097\/MPH.0000000000002506","article-title":"Global Epidemiology of Gaucher Disease: An Updated Systematic Review and Meta-Analysis","volume":"45","author":"Wang","year":"2023","journal-title":"J. Pediatr. Hematol.\/Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1038\/sj.embor.embor873","article-title":"X-ray Structure of Human Acid-Beta-Glucosidase, the Defective Enzyme in Gaucher Disease","volume":"4","author":"Dvir","year":"2003","journal-title":"EMBO Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1016\/S0140-6736(08)61522-6","article-title":"Phenotype, Diagnosis, and Treatment of Gaucher\u2019s Disease","volume":"372","author":"Grabowski","year":"2008","journal-title":"Lancet"},{"key":"ref_5","first-page":"82","article-title":"Management of Gaucher Disease: Enzyme Replacement Therapy","volume":"12","author":"Zimran","year":"2014","journal-title":"Pediatr. Endocrinol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1023\/A:1010335505357","article-title":"Inhibition of Substrate Synthesis as a Strategy for Glycolipid Lysosomal Storage Disease Therapy","volume":"24","author":"Platt","year":"2001","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1016\/j.ymgme.2022.03.001","article-title":"The Diagnosis and Management of Gaucher Disease in Pediatric Patients: Where Do We Go from Here?","volume":"136","author":"Weinreb","year":"2022","journal-title":"Mol. Genet. Metab."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2705","DOI":"10.1021\/jm301557k","article-title":"Pharmacological Chaperones as Therapeutics for Lysosomal Storage Diseases","volume":"56","author":"Boyd","year":"2013","journal-title":"J. Med. Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1740","DOI":"10.1039\/C7SC04712F","article-title":"Tuning Protein Folding in Lysosomal Storage Diseases: The Chemistry behind Pharmacological Chaperones","volume":"9","author":"Pereira","year":"2018","journal-title":"Chem. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1171","DOI":"10.1111\/cbdd.13182","article-title":"Biological Activities of 3,4,5-trihydroxypiperidines and Their N- and O-derivatives","volume":"92","author":"Prichard","year":"2018","journal-title":"Chem. Biol. Drug Des."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5497","DOI":"10.1039\/C6CC01564F","article-title":"Glycomimetic-Based Pharmacological Chaperones for Lysosomal Storage Disorders: Lessons from Gaucher, GM1-Gangliosidosis and Fabry Diseases","volume":"52","author":"Mellet","year":"2016","journal-title":"Chem. Commun."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Mart\u00ednez-Bail\u00e9n, M., Clemente, F., Matassini, C., and Cardona, F. (2022). GCase Enhancers: A Potential Therapeutic Option for Gaucher Disease and Other Neurological Disorders. Pharmaceuticals, 15.","DOI":"10.3390\/ph15070823"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1007\/s40265-019-01090-4","article-title":"Migalastat: A Review in Fabry Disease","volume":"79","author":"McCafferty","year":"2019","journal-title":"Drugs"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1111\/j.1471-4159.1982.tb07950.x","article-title":"Accumulation of Glucosylceramide and Glucosylsphingosine (Psychosine) in Cerebrum and Cerebellum in Infantile and Juvenile Gaucher Disease","volume":"39","author":"Nilsson","year":"1982","journal-title":"J. Neurochem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1023\/A:1015137917508","article-title":"Glycolipid Analysis of Different Tissues and Cerebrospinal Fluid in Type II Gaucher Disease","volume":"25","author":"Gornati","year":"2002","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1016\/j.ymgme.2007.11.011","article-title":"Glucosylceramide Accumulation Is Not Confined to the Lysosome in Fibroblasts from Patients with Gaucher Disease","volume":"93","author":"Fuller","year":"2008","journal-title":"Mol. Genet. Metab."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Vanni, C., Clemente, F., Paoli, P., Morrone, A., Matassini, C., Goti, A., and Cardona, F. (2022). 3,4,5-Trihydroxypiperidine Based Multivalent Glucocerebrosidase (GCase) Enhancers. ChemBioChem, 23.","DOI":"10.1002\/cbic.202200077"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1111\/jnc.13385","article-title":"The Relationship between Glucocerebrosidase Mutations and Parkinson Disease","volume":"139","author":"Schapira","year":"2016","journal-title":"J. Neurochem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.1021\/ja0378207","article-title":"Dissecting the Cholera Toxin\u2212Ganglioside GM1 Interaction by Isothermal Titration Calorimetry","volume":"126","author":"Turnbull","year":"2004","journal-title":"J. Am. Chem. Soc."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"17684","DOI":"10.1038\/s41598-019-53995-5","article-title":"Pre-Diagnosing and Managing Patients with GM1 Gangliosidosis and Related Disorders by the Evaluation of GM1 Ganglioside Content","volume":"9","author":"Tonin","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Capitini, C., Feo, F., Caciotti, A., Tonin, R., Lulli, M., Coviello, D., Guerrini, R., Calamai, M., and Morrone, A. (2022). Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann\u2013Pick Type C. Biomedicines, 10.","DOI":"10.3390\/biomedicines10081962"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Clemente, F., Mart\u00ednez-Bail\u00e9n, M., Matassini, C., Morrone, A., Falliano, S., Caciotti, A., Paoli, P., Goti, A., and Cardona, F. (2022). Synthesis of a New \u03b2-Galactosidase Inhibitor Displaying Pharmacological Chaperone Properties for GM1 Gangliosidosis. Molecules, 27.","DOI":"10.3390\/molecules27134008"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"240","DOI":"10.2350\/09-11-0744-CR.1","article-title":"Gaucher Disease with Prenatal Onset and Perinatal Death Due to Compound Heterozygosity for the Missense R131C and Null Rec Nci I GBA Mutations","volume":"14","author":"Goebl","year":"2011","journal-title":"Pediatr. Dev. Pathol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1398","DOI":"10.1002\/humu.22124","article-title":"Gaucher Disease Paradigm: From ERAD to Comorbidity","volume":"33","author":"Horowitz","year":"2012","journal-title":"Hum. Mutat."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"770","DOI":"10.1016\/j.cell.2007.10.018","article-title":"LIMP-2 Is a Receptor for Lysosomal Mannose-6-Phosphate-Independent Targeting of \u03b2-Glucocerebrosidase","volume":"131","author":"Reczek","year":"2007","journal-title":"Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"570","DOI":"10.1056\/NEJM198703053161002","article-title":"A Mutation in the Human Glucocerebrosidase Gene in Neuronopathic Gaucher\u2019s Disease","volume":"316","author":"Tsuji","year":"1987","journal-title":"N. Engl. J. Med."},{"key":"ref_27","first-page":"212","article-title":"Gaucher Disease: Molecular Heterogeneity and Phenotype-Genotype Correlations","volume":"45","author":"Theophilus","year":"1989","journal-title":"Am. J. Hum. Genet."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1006\/bcmd.1998.0210","article-title":"A Novel Complex Allele and Two New Point Mutations in Type 2 (Acute Neuronopathic) Gaucher Disease","volume":"24","author":"Sinclair","year":"1998","journal-title":"Blood Cells Mol. Dis."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1016\/j.ymgme.2012.04.018","article-title":"Pharmacological Chaperones Facilitate the Post-ER Transport of Recombinant N370S Mutant \u03b2-Glucocerebrosidase in Plant Cells: Evidence That N370S Is a Folding Mutant","volume":"106","author":"Babajani","year":"2012","journal-title":"Mol. Genet. Metab."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1007\/s00401-023-02601-0","article-title":"More than Meets the Eye in Parkinson\u2019s Disease and Other Synucleinopathies: From Proteinopathy to Lipidopathy","volume":"146","author":"Outeiro","year":"2023","journal-title":"Acta Neuropathol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1093\/hmg\/6.2.205","article-title":"Generalized CNS Disease and Massive GM1-Ganglioside Accumulation in Mice Defective in Lysosomal Acid Beta-Galactosidase","volume":"6","author":"Hahn","year":"1997","journal-title":"Hum. Mol. Genet."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"734878","DOI":"10.3389\/fgene.2021.734878","article-title":"GM1 Gangliosidosis-A Mini-Review","volume":"12","author":"Nicoli","year":"2021","journal-title":"Front. Genet."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Breiden, B., and Sandhoff, K. (2020). Mechanism of Secondary Ganglioside and Lipid Accumulation in Lysosomal Disease. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21072566"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1016\/j.bbamcr.2008.11.014","article-title":"Secondary Lipid Accumulation in Lysosomal Disease","volume":"1793","author":"Walkley","year":"2009","journal-title":"Biochim. Biophys. Acta (BBA)\u2014Mol. Cell Res."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/2\/453\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:48:47Z","timestamp":1760104127000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/2\/453"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,17]]},"references-count":34,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["molecules29020453"],"URL":"https:\/\/doi.org\/10.3390\/molecules29020453","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,17]]}}}