The use of human pluripotent stem cells (hPSCs) for in vitro disease modeling is limited by the lack of robust and efficient protocols for the differentiation of relevant adult cell types. Previously, we have reported a method to generate vascular endothelial cells from hPSCs (Patsch et al. Nat Cell Biol. 2015). This novel and robust protocol in conjunction with use of programmed nucleases allows generation of relevant endothelilal cell disease models. We will show examples of modeling high-barrier resistance endothelial cells in vitro, by generation of CLDN5 reporter and use of chemical profiling, and modeling the effect of a severe metabolic genetic disease (AKT2 loss-of-function or dominant active mutation) on endothelial cells.