Fabry Disease-Associated Cardiovascular Phenotypes Using Isogenic GLA-Knockout Human iPSCs
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Fabry disease is an X-linked lysosomal storage disorder caused by mutations in the GLA gene, leading to deficient α-galactosidase A activity and subsequent accumulation of globotriaosylceramide (Gb3). This accumulation contributes to progressive multi-organ dysfunction, with cardiovascular complications- particularly endothelial dysfunction and left ventricular hypertrophy- being major drivers of disease morbidity and mortality. Although enzyme replacement therapy (ERT) is currently the standard treatment, its effectiveness is limited in addressing advanced cardiovascular pathology. To better understand Fabry-associated vascular and cardiac phenotypes, we developed an isogenic human induced pluripotent stem cell (hiPSC) model in which GLA was knocked out using CRISPR/Cas9. GLA-knockout (GLA-KO) hiPSCs were differentiated into endothelial cells (ECs) and cardiomyocytes (CMs) to evaluate disease-relevant phenotypes in vitro. GLA-KO ECs exhibited normal morphology and differentiation capacity but showed markedly impaired tube formation, elevated inflammatory gene expression (ICAM1, VCAM1, SELE), and increased mitochondrial and cytoplasmic reactive oxygen species (ROS) levels. GLA-KO-CMs demonstrated enlarged cell size and nuclear translocation of NFATC4, consistent with hypertrophic remodeling. Together, these findings recapitulate key features of Fabry vasculopathy and cardiomyopathy in a genetically defined, human-derived system. This platform enables direct investigation of Gb3-induced oxidative and inflammatory mechanisms and provides a valuable model for preclinical evaluation of therapeutic strategies targeting cardiovascular manifestations of Fabry disease.
Abstract
Fabry disease is an X-linked lysosomal storage disorder caused by mutations in the GLA gene, leading to deficient α-galactosidase A activity and subsequent accumulation of globotriaosylceramide (Gb3). This accumulation contributes to progressive multi-organ dysfunction, with cardiovascular complications- particularly endothelial dysfunction and left ventricular hypertrophy- being major drivers of disease morbidity and mortality. Although enzyme replacement therapy (ERT) is currently the standard treatment, its effectiveness is limited in addressing advanced cardiovascular pathology. To better understand Fabry-associated vascular and cardiac phenotypes, we developed an isogenic human induced pluripotent stem cell (hiPSC) model in which GLA was knocked out using CRISPR/Cas9. GLA-knockout (GLA-KO) hiPSCs were differentiated into endothelial cells (ECs) and cardiomyocytes (CMs) to evaluate disease-relevant phenotypes in vitro. GLA-KO ECs exhibited normal morphology and differentiation capacity but showed markedly impaired tube formation, elevated inflammatory gene expression (ICAM1, VCAM1, SELE), and increased mitochondrial and cytoplasmic reactive oxygen species (ROS) levels. GLA-KO-CMs demonstrated enlarged cell size and nuclear translocation of NFATC4, consistent with hypertrophic remodeling. Together, these findings recapitulate key features of Fabry vasculopathy and cardiomyopathy in a genetically defined, human-derived system. This platform enables direct investigation of Gb3-induced oxidative and inflammatory mechanisms and provides a valuable model for preclinical evaluation of therapeutic strategies targeting cardiovascular manifestations of Fabry disease.