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Ryan et al. J Transl Genet Genom. 2025;9:48-61 Journal of Translational
DOI: 10.20517/jtgg.2024.87
Genetics and Genomics

Original Article Open Access

Investigating lysosomal dysfunction in Fabry
disease using induced pluripotent stem cell-derived

podocytes

1
1
1
2
3
1
Caitlin R. Ryan , Andrea F. Wise , Elisha Tindoy , Shoni Bruell , Maria Fuller , Kathleen M. Nicholls ,
Sharon D. Ricardo 1
1
Department of Pharmacology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria 3800, Australia.
2
Genetics and Molecular Pathology, South Australia Pathology at Women's and Children's Hospital and Adelaide Medical School
and School of Biological Sciences, University of Adelaide, Adelaide, SA 5006, Australia.
3
Department of Nephrology, The Royal Melbourne Hospital and Department of Medicine (RMH), University of Melbourne,
Parkville 3000, Australia.
Correspondence to: Prof. Sharon D. Ricardo, Department of Pharmacology, Biomedicine Discovery Institute, Monash University,
9 Ancora Imparo Way, Clayton, Victoria 3800, Australia. E-mail: sharon.ricardo@monash.edu
How to cite this article: Ryan CR, Wise AF, Tindoy E, Bruell S, Fuller M, Nicholls KM, Ricardo SD. Investigating lysosomal
dysfunction in Fabry disease using induced pluripotent stem cell-derived podocytes. J Transl Genet Genom. 2025;9:48-61. https:/
/dx.doi.org/10.20517/jtgg.2024.87
Received: 25 Oct 2024 First Decision: 20 Jan 2025 Revised: 3 Feb 2025 Accepted: 12 Feb 2025 Published: 26 Feb 2025

Academic Editor: Sanjay Gupta Copy Editor: Fangling Lan Production Editor: Fangling Lan

Abstract
Aims: This study used induced pluripotent stem cell-derived podocytes from a Fabry disease (FD) patient carrying
the p.Met284Thr pathogenic variant as an in vitro model to investigate lysosomal abnormalities driving cell
pathology. Proteomic analysis was used to assess changes in lysosomal protein abundance in FD podocytes
compared to controls. Additionally, temporal changes in lysosome number in FD podocytes were analyzed using
automated live-cell imaging.

Methods: Label-free mass spectrometry proteomics was performed on FD podocytes at day 10 of differentiation
compared to controls. For live-cell imaging, cultured podocytes were transfected with CellLight Lysosomes-GFP
and Plasma Membrane-CFP, and then visualized and quantified on days 10 and 20 post-differentiation using a
Perkin Elmer Phenix High Content Screening Microscope.

Results: Proteomic analysis showed dysregulation of glycosphingolipid metabolism proteins, including decreased
galactosidase alpha (GLA; P < 0.01) and increased galactosylceramidase and glucosylceramidase (P < 0.01) in FD

© The Author(s) 2025. Open Access This article is licensed under a Creative Commons Attribution 4.0
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adaptation, distribution and reproduction in any medium or format, for any purpose, even commercially, as
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indicate if changes were made.

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