Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann-Pick Type C

Year: 2022

Authors: Capitini C.; Feo F.; Caciotti A.; Tonin R.; Lulli M.; Coviello D.; Guerrini R.; Calamai M.; Morrone A.

Autors Affiliation: Univ Florence, European Lab Nonlinear Spect LENS, I-50019 Sesto Fiorention, Italy; Natl Inst Opt Natl Res Council CNR INO, I-50019 Sesto Fiorentino, Italy; Meyer Childrens Hosp, Neurosci Dept, Lab Mol Biol Neurometab Dis, Viale Pieraccini 24, I-50139 Florence, Italy; Univ Florence, Dept Expt & Clin Biomed Sci Mario Serio, I-50134 Florence, Italy; IRCCS Ist Giannina Gaslini, Lab Human Genet, I-16147 Genoa, Italy; Univ Florence, Dept Neurosci Psychol Pharmacol & Child Hlth, I-50121 Florence, Italy.

Abstract: Background: Early diagnosis is essential in the field of lysosomal storage disorders for the proper management of patients and for starting therapies before irreversible damage occurs, particularly in neurodegenerative conditions. Currently, specific biomarkers for the diagnosis of lysosomal storage disorders are lacking in routine laboratory practice, except for enzymatic tests, which are available only in specialized metabolic centers. Recently, we established a method for measuring and verifying changes in GM1 ganglioside levels in peripheral blood lymphocytes in patients with GM1 gangliosidosis. However, fresh blood is not always available, and using frozen/thawed lymphocytes can lead to inaccurate results. Methods: We used frozen/thawed fibroblasts obtained from stored biopsies to explore the feasibility of fluorescent imaging and flow-cytometric methods to track changes in storage materials in fibroblasts from patients with three lysosomal neurodegenerative conditions: GM1 gangliosidosis, Sialidosis, and Niemann-Pick type C. We used specific markers for each pathology. Results and Conclusions: We demonstrated that with our methods, it is possible to clearly distinguish the levels of accumulated metabolites in fibroblasts from affected and unaffected patients for all the three pathologies considered. Our methods proved to be rapid, sensitive, unbiased, and potentially applicable to other LSDs.

Journal/Review: BIOMEDICINES

Volume: 10 (8)      Pages from: 1962-1  to: 1962-12

More Information: The research was funded by Regione Toscana (Bando Salute 2018, DD15397) for the project Lysolate and by European Union´s Horizon 2020 research and innovation programme under grant agreement No 654148 Laserlab-Europe.
M Calamai and A Morrone, equal last author contribution
KeyWords: GM1 gangliosidosis; sialidosis; Niemann-Pick type C; lysosomal storage disorders; flow cytometry; fluorescent imaging; Cholera Toxin B; Filipin; cholesterol; sialic acid; biomarkers
DOI: 10.3390/biomedicines10081962

Citations: 3
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