Department II of Internal Medicine Medicine and Center for Rare Diseases Cologne, University Hospital of Cologne, Cologne, Germany.
Cologne Excellence Cluster on Cellular Stress Responses in Ageing-Associated Diseases (CECAD), University of Cologne, Cologne, Germany.
Sci Rep. 2018 Jul 23;8(1):11042. doi: 10.1038/s41598-018-29240-w.
Fabry disease is a lysosomal storage disorder resulting from impaired alpha-galactosidase A (α-Gal A) enzyme activity due to mutations in the GLA gene. Currently, powerful diagnostic tools and in vivo research models to study Fabry disease are missing, which is a major obstacle for further improvements in diagnosis and therapy. Here, we explore the utility of urine-derived primary cells of Fabry disease patients. Viable cells were isolated and cultured from fresh urine void. The obtained cell culture, modeling the renal epithelium, is characterized by patient-specific information. We demonstrate that this non-invasive source of patient cells provides an adequate cellular in vivo model as cells exhibit decreased α-Gal A enzyme activity and concomitant globotriaosylceramide accumulation. Subsequent quantitative proteomic analyses revealed dysregulation of endosomal and lysosomal proteins indicating an involvement of the Coordinated Lysosomal Expression and Regulation (CLEAR) network in the disease pathology. This proteomic pattern resembled data from our previously described human podocyte model of Fabry disease. Taken together, the employment of urine-derived primary cells of Fabry disease patients might have diagnostic and prognostic implications in the future. Our findings pave the way towards a more detailed understanding of pathophysiological mechanisms and may allow the development of future tailored therapeutic strategies.
法布里病是一种溶酶体贮积病，由于 GLA 基因突变导致α-半乳糖苷酶 A（α-Gal A）酶活性受损而引起。目前，缺乏强大的诊断工具和用于研究法布里病的体内研究模型，这是进一步改善诊断和治疗的主要障碍。在这里，我们探讨了利用法布里病患者的尿液来源原代细胞的可能性。我们从新鲜的尿液中分离并培养了有活力的细胞。获得的细胞培养物模拟了肾上皮，具有患者特异性信息。我们证明，这种非侵入性的患者细胞来源为细胞提供了充足的体内模型，因为细胞表现出 α-Gal A 酶活性降低和伴随的Globotriaosylceramide 积累。随后的定量蛋白质组学分析显示内体和溶酶体蛋白的失调，表明协调溶酶体表达和调节（CLEAR）网络参与疾病病理学。这种蛋白质组模式与我们之前描述的法布里病人类足细胞模型的数据相似。总之，法布里病患者尿液来源的原代细胞的应用可能具有未来的诊断和预后意义。我们的发现为深入了解病理生理机制铺平了道路，并可能允许未来开发出有针对性的治疗策略。
