Fabry disease (FD) is a rare inherited disorder characterized by a wide range of systemic\nsymptoms; it is particularly associated with cardiovascular and renal problems. Enzyme\nreplacement therapy and pharmacological chaperone migalastat are the only approved and\neffective treatment strategies for FD patients. It is well documented that alpha-galactosidase A\n(GLA) enzyme activity deficiency causes globotriaosylceramide (Gb3) accumulation, which plays a\ncrucial role in the etiology of FD. However, the detailed mechanisms remain unclear, and the lack\nof a reliable and powerful disease model is an obstacle. In this study, we created such a model by\nusing CRISPR/Cas9-mediated editing of GLA gene to knockout its expression in human embryonic\nstem cells (hESCs). The cardiomyocytes differentiated from these hESCs (GLA-null CMs) were\ncharacterized by the accumulation of Gb3 and significant increases of cell surface area, the\nlandmarks of FD-associated cardiomyopathy. Furthermore, we used mass spectrometry to compare\nthe proteomes of GLA-null CMs and parental wild type CMs and found that the Rab GTPases\ninvolved in exocytotic vesicle release were significantly downregulated. This caused impairment of\nautophagic flux and protein turnover, resulting in an increase of reactive oxygen species and\napoptosis. To summarize, we established a FD model which can be used as a promising tool to study\nhuman hypertrophic cardiomyopathy in a physiologically and pathologically relevant manner and\nto develop new therapies by targeting Rab GTPases signaling-related exosomal vesicles\ntransportation.
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