Enhanced thrombospondin-1 causes dysfunction of vascular endothelial cells derived from Fabry disease-induced pluripotent stem cells

Abstract

Background: Fabry disease (FD) is a recessive X-linked lysosomal storage disorder caused by a-galactosidase A (GLA) deficiency. Although the mechanism is unclear, GLA deficiency causes an accumulation of globotriaosylceramide (Gb3), leading to vasculopathy. Methods: To explore the relationship between the accumulation of Gb3 and vasculopathy, induced pluripotent stem cells generated from four Fabry patients (FD-iPSCs) were differentiated into vascular endothelial cells (VECs). Genome editing using CRISPR-Cas9 system was carried out to correct the GLA mutation or to delete Thrombospondin-1 (TSP-1). Global transcriptomes were compared between wild-type (WT)-and FD-VECs by RNA-sequencing analysis. Findings: Here, we report that overexpression of TSP-1 contributes to the dysfunction of VECs in FD. VECs originating from FD-iPSCs (FD-VECs) showed aberrant angiogenic functionality even upon treatment with recombinant a-galactosidase. Intriguingly, FD-VECs produced more p-SMAD2 and TSP-1 than WT-VECs. We also found elevated TSP-1 in the peritubular capillaries of renal tissues biopsied from FD patients. Inhibition of SMAD2 signaling or knock out of TSP-1 (TSP-1 À/À ) rescues normal vascular functionality in FD-VECs, like in gene-corrected FD-VECs. In addition, the enhanced oxygen consumption rate is reduced in TSP-1 À/À FD-VECs. Interpretation: The overexpression of TSP-1 secondary to Gb3 accumulation is primarily responsible for the observed FD-VEC dysfunction. Our findings implicate dysfunctional VEC angiogenesis in the peritubular capillaries in some of the complications of Fabry disease.

Biomarkers in Fabry Disease. Implications for Clinical Diagnosis and Follow-up

“…Increased reactive oxygen species (ROS) and decreased nitric oxide (NO) also contribute to endothelial damage [24][25][26]. A different hypothesis about the development of vasculopathy, proposes that the accumulation of endothelial Gb3 causes a dysregulation of the enzyme nitric oxide endothelial synthase (eNOS) with the consequent formation of oxidant species derived from NO, which could be direct markers of vasculopathy in FD [27][28][29][30][31][32][33][34]. In addition, Gb3 could also contribute to endothelial damage from other different pathogenic mechanisms related to KCa3.1 channel dysfunction [35,36].…”

Ion channels and pain in Fabry disease

“…The field of FD research has also benefited from the advent of iPSCs [55][56][57][58] and the CRISPR/CAS9-mediated genome engineering technology. 59,60 This technique facilitates site-specific DNA deletions, insertions, inversions, and replacements and thus shows therapeutic potential, and is an invaluable tool in establishing the causal relationship between genes and stem cell behavior. 64 However, as the pain experience results from integrated pathways, all the way from sensory transduction in the periphery to perception in the brain, it is critical to study the system with each level of processing intact.…”

LRP-1 Matricellular Receptor Involvement in Triple Negative Breast Cancer Tumor Angiogenesis

“…In THBS1 up-regulated cells, the secreted VEGF could be sequestrated, and is thus not sufficient for the cells to ensure a proper VEGF-stimulated angiogenesis. As THBS1 regulates vessel stabilization, its overexpression has been shown to suppress vascular growth and expand vessel diameter [58], suggesting that it could be associated with dysfunctional angiogenesis, like in Fabry disease [59]. Despite an increased plasminogen expression and one of its activators in shLRP-1 TCM, a decreased plasmin activity was measured.…”