Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann–Pick Type C
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...
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2022-08-01
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| author | Claudia Capitini Federica Feo Anna Caciotti Rodolfo Tonin Matteo Lulli Domenico Coviello Renzo Guerrini Martino Calamai Amelia Morrone |
| author_facet | Claudia Capitini Federica Feo Anna Caciotti Rodolfo Tonin Matteo Lulli Domenico Coviello Renzo Guerrini Martino Calamai Amelia Morrone |
| author_sort | Claudia Capitini |
| collection | DOAJ |
| description | 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. |
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| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-09T11:50:04Z |
| publishDate | 2022-08-01 |
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| spelling | doaj.art-08675c6fef3049d99872a434a65b9d872023-11-30T23:15:11ZengMDPI AGBiomedicines2227-90592022-08-01108196210.3390/biomedicines10081962Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann–Pick Type CClaudia Capitini0Federica Feo1Anna Caciotti2Rodolfo Tonin3Matteo Lulli4Domenico Coviello5Renzo Guerrini6Martino Calamai7Amelia Morrone8European Laboratory for Non-Linear Spectroscopy (LENS), University of Florence, 50019 Sesto-Fiorention, ItalyLaboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children’s Hospital, Viale Pieraccini n. 24, 50139 Florence, ItalyLaboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children’s Hospital, Viale Pieraccini n. 24, 50139 Florence, ItalyLaboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children’s Hospital, Viale Pieraccini n. 24, 50139 Florence, ItalyDepartment of Experimental and Clinical Biomedical Sciences “Mario Serio”, University of Florence, 50134 Florence, ItalyLaboratory of Human Genetics, IRCCS Istituto Giannina Gaslini, 16147 Genoa, ItalyLaboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children’s Hospital, Viale Pieraccini n. 24, 50139 Florence, ItalyEuropean Laboratory for Non-Linear Spectroscopy (LENS), University of Florence, 50019 Sesto-Fiorention, ItalyLaboratory of Molecular Biology of Neurometabolic Diseases, Neuroscience Department, Meyer Children’s Hospital, Viale Pieraccini n. 24, 50139 Florence, ItalyBackground: 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.https://www.mdpi.com/2227-9059/10/8/1962GM1 gangliosidosissialidosisNiemann–Pick type Clysosomal storage disordersflow cytometryfluorescent imaging |
| spellingShingle | Claudia Capitini Federica Feo Anna Caciotti Rodolfo Tonin Matteo Lulli Domenico Coviello Renzo Guerrini Martino Calamai Amelia Morrone Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann–Pick Type C Biomedicines GM1 gangliosidosis sialidosis Niemann–Pick type C lysosomal storage disorders flow cytometry fluorescent imaging |
| title | Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann–Pick Type C |
| title_full | Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann–Pick Type C |
| title_fullStr | Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann–Pick Type C |
| title_full_unstemmed | Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann–Pick Type C |
| title_short | Fluorescent In Situ Staining and Flow Cytometric Procedures as New Pre-Diagnostic Tests for Sialidosis, GM1 Gangliosidosis and Niemann–Pick Type C |
| title_sort | fluorescent in situ staining and flow cytometric procedures as new pre diagnostic tests for sialidosis gm1 gangliosidosis and niemann pick type c |
| topic | GM1 gangliosidosis sialidosis Niemann–Pick type C lysosomal storage disorders flow cytometry fluorescent imaging |
| url | https://www.mdpi.com/2227-9059/10/8/1962 |
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