Human kidney organoids reveal the role of glutathione in Fabry disease

Abstract

Fabry disease is an X-linked lysosomal storage disease caused by a mutation in the galactosidase alpha (GLA) gene. Despite advances in therapeutic technologies, the lack of humanized experimental models of Fabry disease has limited the development of new therapies to cure the disease. Herein, we modeled Fabry disease using human inducible pluripotent stem cell (iPSC)-derived kidney organoids and the CRISPR–Cas9 genome-editing system. GLA-mutant human kidney organoids revealed deformed podocytes and tubular cells with accumulation of globotriaosylceramide (Gb3). Ultrastructural analysis showed abundant electron-dense granular deposits and electron-dense lamellate lipid-like deposits that formed concentric bodies (zebra bodies) in the cytoplasm of podocytes and tubules. The oxidative stress level was increased in GLA-mutant kidney organoids, and the increase was accompanied by apoptosis. Enzyme replacement treatment (ERT) with recombinant human α-Gal A decreased the Gb3 accumulation and oxidative stress, which resulted in amelioration of the deformed cellular structure of the GLA-mutant kidney organoids. Transcription profile analyses showed decreased glutathione (GSH) metabolism in GLA-mutant kidney organoids. GSH replacement treatment decreased oxidative stress and attenuated the structural deformity of the GLA-mutant kidney organoids. GSH treatment also increased the expression of podocyte and tubular markers and decreased apoptosis. In conclusion, GLA-mutant kidney organoids derived from human iPSCs are valuable tools for studying the mechanisms and developing novel therapeutic alternatives for Fabry disease.

The Effect of Green Tea as an Adjuvant to Enzyme Replacement Therapy on Oxidative Stress in Fabry Disease: A Pilot Study

“…In another study, ascorbate, a potent antioxidant, was found to decrease cerebral hyperperfusion in FD patients on ERT (41). Recently, Kim et al showed in an in vitro disease model of renal FD that treatment with the antioxidant GSH reduced OS and attenuated the structural alterations of the GLA-mutant kidney organoids compared to wild-type kidney organoids (42). Our data support and provide the mechanistic rationale to these studies which suggest that antiinflammatory/antioxidative dietary patterns might be added to FD ERT treatment to contribute to slowing disease progression.…”

Generation of GLA-knockout human embryonic stem cell lines to model peripheral neuropathy in Fabry disease

“…However, although this model has many neuronal features, it is not directly derived from peripheral neurons. Several groups have reported the development of induced pluripotent cell lines using primary cells from Fabry patients [ [23] , [24] , [25] , [26] , [27] , [28] , [29] , [30] ] and the use of CRISPR-Cas9 gene editing either to generate GLA -knockout stem cell models from the hESC cell line WA09 (H9) [ 47 ] and from normal iPSC lines [ [48] , [49] , [50] ], or to generate gene-corrected Fabry iPSC [ 50 , 51 ] to create isogenic controls. To date these pluripotent stem cell lines have been used mainly to study molecular mechanisms of cardiomyopathy found in Fabry patients [ 23 , 28 , 30 , 47 , 50 , 52 ], in addition to kidney [ 49 ] and vascular endothelial dysfunction [ 51 ], but to our knowledge there have been no reports of using Fabry pluripotent cells to model peripheral pain-sensing neurons.…”

Fabry disease: Mechanism and therapeutics strategies

“…generated GLA -mutant kidney organoids via CRISPR-Cas9 technology. Transmission electron microscopy and oil red O staining analysis showed greater lipid accumulation in Fabry disease model organoids compared with that in the controls ( Kim et al, 2021 ), suggesting that lipid metabolism may have been inhibited in the model organoids. However, the mechanism involved in lipid droplet formation in Fabry disease remains elusive, even in preclinical studies.…”