Enzyme Replacement Therapy Clears Gb3 Deposits from a Podocyte Cell Culture Model of Fabry Disease but Fails to Restore Altered Cellular Signaling

Cell Physiol Biochem. 2019;52(5):1139-1150. doi: 10.33594/000000077.

Abstract

Background/aims: Fabry disease (FD) is a lysosomal storage disorder characterized by impaired alpha-galactosidase A (α-Gal A) enzyme activity due to mutations in the GLA gene. While virtually all tissues are affected, renal damage is particularly critical for the patients' outcome. Currently, powerful diagnostic tools and in vivo research models to study FD in the kidney are lacking, which is a major obstacle for further improvements in diagnosis and therapy. The present study focuses on the effects of enzyme replacement therapy on a previously established podocyte cell culture model of Fabry disease.

Methods: We investigated the effect of in vitro application of α-Gal A on Fabry podocytes for 3 days, mimicking enzyme replacement therapy. We studied reduction of Gb3 levels and dysregulated molecular pathways such as autophagy, mTOR/AKT signaling and pro-fibrotic signaling by employing immunofluorescence, electron microscopy, tandem mass spectrometry and western blot.

Results: We detected complete resolution of Gb3 accumulation in Fabry podocytes upon α-Gal A treatment. Despite robust Gb3 clearance, dysregulation of the signaling pathways investigated was not reversed.

Conclusion: This study presents first evidence for Gb3-independent effects regarding dysregulation of signal transduction mechanisms in FD not recovering upon α-Gal A treatment. We assume that intracellular alterations observed in FD may have a point of no return after which a reversal of dysregulated cellular signal transduction by α-Gal A treatment is not effective, despite Gb3 clearance. Our observations suggest further research on signal transduction mechanisms altered in Fabry podocytes and on determining the appropriate time for initiation of Fabry therapy.

Keywords: Enzyme Replacement Therapy; Fabry; Podocyte.

MeSH terms

  • Cell Culture Techniques
  • Cell Line, Transformed
  • Enzyme Replacement Therapy*
  • Fabry Disease* / drug therapy
  • Fabry Disease* / metabolism
  • Fabry Disease* / pathology
  • Humans
  • Models, Biological*
  • Podocytes / metabolism*
  • Podocytes / pathology
  • Signal Transduction / drug effects*
  • Trihexosylceramides / metabolism*
  • alpha-Galactosidase / therapeutic use*

Substances

  • Trihexosylceramides
  • globotriaosylceramide
  • alpha-Galactosidase