In this study, we characterized dia-hiPSC-ECs and found that they not only recapitulated several phenotypes of endothelial dysfunction, such as increased vasoconstrictor (endothelin-1) and adhesion molecule (ICAM-1) production and impaired angiogenesis, but also showed disrupted glycine homeostasis, increased senescence, and impaired mitochondrial function. Here, EDN1 is linked to endothelial dysfunction.