Urine-derived cells: a promising diagnostic tool in Fabry disease patients

Gisela G Slaats,Bernhard Schermer,Christine E Kurschat,Martin Hoehne,Linda Blomberg,Markus M Rinschen,Laura E Frech,Fabian Braun,Thomas Benzing

doi:10.1038/s41598-018-29240-w

Abstract

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.

Highlights

Fabry disease [MIM: 301500] is a hereditary disorder of the glycosphingolipid metabolism caused by mutations in the alpha-galactosidase A (GLA) gene located on the X chromosome
We describe the use of urine-derived primary cells of Fabry disease patients as an effective tool for diagnosis and research of pathomechanisms
In the three samples obtained from male patients no α-Galactosidase A (α-Gal) A activity could be detected, in contrast to residual enzyme activity measured in female patient samples (Supplementary Fig. S1E)

Summary

Introduction

Fabry disease [MIM: 301500] is a hereditary disorder of the glycosphingolipid metabolism caused by mutations in the alpha-galactosidase A (GLA) gene located on the X chromosome. Research has focused on the development of new biomarkers for diagnosis and for monitoring disease course and treatment efficacy. The portfolio of routine biomarkers and disease models is still very limited For these reasons, we propose to introduce primary urine cells of Fabry disease patients as a diagnostic tool. Gb3 derivative globotriaosylsphingosine, is a potential tool for monitoring treatment efficacy in Fabry disease patients[5,6]. We describe the use of urine-derived primary cells of Fabry disease patients as an effective tool for diagnosis and research of pathomechanisms. In cultured urine-derived primary cells of Fabry disease patients we detected decreased α-Gal A enzyme activity and concomitant Gb3 accumulation. For future studies on disease mechanisms and therapeutic interventions, viable urine-derived Fabry disease cells could present a valuable tool

Methods

Results

Conclusion