Characterization of cellular phenotypes in neurons derived from induced pluripotent stem cells of male patients with Fabry disease

Abstract

Fabry disease (FD) is an X-linked inherited lysosomal metabolism disorder in which globotriaosylceramide (Gb3) accumulates in various organs resulting from a deficiency in alpha-galactosidase A. The clinical features of FD include progressive impairments of the renal, cardiac, and peripheral nervous systems. In addition, patients with FD often develop neuropsychiatric symptoms, such as depression and dementia, which are believed to be induced by the cellular injury of cerebrovascular and partially neuronal cells due to Gb3 accumulation. Although the analysis of autopsy brain tissue from patients with FD showed no accumulation of Gb3, abnormal deposits of Gb3 were found in the neurons of several brain areas, including the hippocampus. Therefore, in this study, we generated induced pluripotent stem cells (iPSCs) from patients with FD and differentiated them into neuronal cells to investigate pathological and biological changes in the neurons of FD. Neural stem cells (NSCs) and neurons were successfully differentiated from the iPSCs we generated; however, cellular damage and morphological changes were not found in these cells. Immunostaining revealed no Gb3 accumulation in NSCs and neurons. Transmission electron microscopy did not reveal any zebra body-like structures or inclusion bodies, which are characteristic of FD. These results indicated that neuronal cells derived from FD-iPSCs exhibited normal morphology and no Gb3 accumulation. It is likely that more in vivo environment-like cultures are needed for iPSC-derived neurons to reproduce disease-specific features.

Small fibre neuropathy in Fabry disease: a human-derived neuronal in vitro disease model and pilot data

“…In vitro models have been generated for genetic pain syndromes using iPSC-derived sensory neurons 15-17 , 43 and have helped to unravel disease pathophysiology 15 , 43 , 44 and identify novel targets for treatment. 16 , 17 In Fabry research, generation of sensory neurons retaining cellular disease phenotype using patient biomaterial was not successful so far, 45 , 46 while differentiation of Fabry iPSC to cardiomyocytes, 11 , 47 podocytes 48 or vascular endothelial cells 49 was achieved. We modified a published differentiation strategy 31 and generated iPSC-derived sensory neurons showing pathognomonic Gb3 accumulations.…”

Small fibre neuropathy in Fabry disease: a human-derived neuronalin vitrodisease model

“…In vitro models have been generated for genetic pain syndromes using iPSC-derived sensory neurons [15][16][17]44 and have helped to unravel disease pathophysiology 15,44,45 and identify novel targets for treatment. 16,17 In Fabry research, generation of sensory neurons retaining cellular disease phenotype using patient biomaterial was not successful so far, 46,47 while differentiation of Fabry iPSC to cardiomyocytes, 11,48 podocytes, 49 or vascular endothelial cells 50 was achieved. We modified a published differentiation strategy 32 and generated iPSCderived sensory neurons showing pathognomonic Gb3 accumulations.…”

Fabry Disease and Central Nervous System Involvement: From Big to Small, from Brain to Synapse

“…In spite of the success of this technology in other LDs [ 65 ], a proper model for FD neurons was still not obtained. Miyajima et al were able to successfully derive NSCs from an FD patient’s iPSCs; however, cellular damage and morphological changes similar to the ones found in the patient’s brain were not recapitulated in these cells, where no Gb3 accumulation was detected [ 66 ].…”