Identification of GM1-Ganglioside Secondary Accumulation in Fibroblasts from Neuropathic Gaucher Patients and Effect of a Trivalent Trihydroxypiperidine Iminosugar Compound on Its Storage Reduction

Year: 2024

Authors: Ceni C., Clemente F., Mangiavacchi F., Matassini C., Tonin R., Caciotti A., Feo F., Coviello D., Morrone A., Cardona F., Calamai M.

Autors Affiliation: Univ Florence, Dept Chem U Schiff DICUS, Via Lastruccia 3-13, FI-50019 Sesto Fiorentino, Italy; Univ Florence, European Lab Nonlinear Spect LENS, I-50019 Sesto Fiorentino, Italy; Meyer Childrens Hosp IRCCS, Neurosci Dept, Lab Mol Biol Neurometab Dis, I-50139 Florence, Italy; IRCCS Ist Giannina Gaslini, Lab Human Genet, I-16147 Genoa, Italy; Univ Florence, Dept Neurosci Psychol Drug Res & Child Hlth NEUROF, I-50121 Florence, Italy; CNR, INO, I-50019 Sesto Fiorentino, Italy.

Abstract: 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 beta-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 beta-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.

Journal/Review: MOLECULES

Volume: 29 (2)      Pages from: 453-1  to: 453-12

More Information: We gratefully thank the AMMeC (Associazione Malattie Metaboliche e Congenite, Italia) and the Cell Line and DNA Biobank from Patients Affected by Genetic Diseases, member of the Telethon Network of Genetic Biobanks funded by Telethon Italy, for providing the fibroblasts.
KeyWords: metabolic disorders; lysosome; glucocerebrosidase; glucosylceramide; GM1; flow cytometry; pharmacological chaperones
DOI: 10.3390/molecules29020453

Citations: 1
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