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.
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.
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.
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.
摘要：
背景：在溶酶体贮积症领域，早期诊断对于正确管理患者和在不可逆损伤发生之前开始治疗至关重要，特别是在神经退行性疾病中。目前,用于诊断溶酶体贮积症的特异性生物标志物在常规实验室实践中缺乏,除了酶测试，它们仅在专门的代谢中心可用。最近,我们建立了一种测量和验证GM1神经节苷脂病患者外周血淋巴细胞GM1神经节苷脂水平变化的方法。然而,新鲜血液并不总是可用的，使用冷冻/解冻的淋巴细胞会导致不准确的结果。
方法：我们使用从保存的活检组织中获得的冷冻/解冻的成纤维细胞来探索荧光成像和流式细胞术方法追踪三种溶酶体神经退行性疾病患者成纤维细胞中保存材料变化的可行性：GM1神经节，唾液中毒,和Niemann-Pick型C。我们对每种病理使用了特定的标记。
结论：我们证明了用我们的方法，对于所考虑的所有三种病理，有可能清楚地区分成纤维细胞与受影响和未受影响的患者中积累的代谢物水平。我们的方法被证明是快速的，敏感,没有偏见,并可能适用于其他LSD。
方法：我们使用从保存的活检组织中获得的冷冻/解冻的成纤维细胞来探索荧光成像和流式细胞术方法追踪三种溶酶体神经退行性疾病患者成纤维细胞中保存材料变化的可行性：GM1神经节，唾液中毒,和Niemann-Pick型C。我们对每种病理使用了特定的标记。
结论：我们证明了用我们的方法，对于所考虑的所有三种病理，有可能清楚地区分成纤维细胞与受影响和未受影响的患者中积累的代谢物水平。我们的方法被证明是快速的，敏感,没有偏见,并可能适用于其他LSD。
