High-throughput identification of small molecules that affect human embryonic vascular development

Abstract

Birth defects, which are in part caused by exposure to environmental chemicals and pharmaceutical drugs, affect 1 in every 33 babies born in the United States each year. The current standard to screen drugs that affect embryonic development is based on prenatal animal testing; however, this approach yields low-throughput and limited mechanistic information regarding the biological pathways and potential adverse consequences in humans. To develop a screening platform for molecules that affect human embryonic development based on endothelial cells (ECs) derived from human pluripotent stem cells, we differentiated human pluripotent stem cells into embryonic ECs and induced their maturation under arterial flow conditions. These cells were then used to screen compounds that specifically affect embryonic vasculature. Using this platform, we have identified two compounds that have higher inhibitory effect in embryonic than postnatal ECs. One of them was fluphenazine (an antipsychotic), which inhibits calmodulin kinase II. The other compound was pyrrolopyrimidine (an antiinflammatory agent), which inhibits vascular endothelial growth factor receptor 2 (VEGFR2), decreases EC viability, induces an inflammatory response, and disrupts preformed vascular networks. The vascular effect of the pyrrolopyrimidine was further validated in prenatal vs. adult mouse ECs and in embryonic and adult zebrafish. We developed a platform based on human pluripotent stem cell-derived ECs for drug screening, which may open new avenues of research for the study and modulation of embryonic vasculature.high-throughput screening | endothelial cells | vascular toxicity | pluripotent stem cells | embryonic endothelial markers

Inflammatory Responses and Barrier Function of Endothelial Cells Derived from Human Induced Pluripotent Stem Cells

“…Nevertheless, despite their potential utility, primary ECs are still preferred to hiPSC-ECs in vascular research and assays, likely due to apparent differences in their developmental and differentiation states. hiPSC-ECs are indeed more similar to embryonic ECs, based on their marker and gene expression profiles ( Orlova et al., 2014a , Rufaihah et al., 2013 , Vazão et al., 2017 ). However, this can have advantages for certain applications, such as screening for embryonic vascular toxicity ( Vazão et al., 2017 ), and perhaps modeling tumor vasculature, since it is also considered immature.…”

The regeneration of macro‐porous electrospun poly(ɛ‐caprolactone) vascular graft during long‐termin situimplantation

“…The endothelialization is essential for the long‐term patency of the vascular graft. In this work, we used vWF as endothelial cell's marker which has been used to detect the endothelial cells in many reports . However, vWF is also expressed in platelets except endothelial cells, not as specific as CD31.…”

iPSCs-based generation of vascular cells: reprogramming approaches and applications

“…A platform based on endothelial cells derived from iPSCs generated from human cord blood was established for drug screening from Vazao et al, which may open new avenues of research for the study and modulation specifically of the embryonic vasculature [ 124 ]. Herein, generated iPSCs were differentiated into embryonic endothelial cells, allowed to mature under flow conditions for more accurate toxicological assessment prior exposing the cells in static conditions to a Library of Pharmacologically Active Compounds (LOPAC, from Sigma).…”

“…Interestingly, two compounds that had higher inhibitory effect in embryonic than postnatal endothelial cells were identified: fluphenazine (an antipsychotic), which inhibits calmodulin kinase II and pyrrolopyrimidine (an anti-inflammatory agent), which inhibits VEGFR2, decreases endothelial cell viability, induces an inflammatory response, and disrupts preformed vascular networks. The vascular effect of the pyrrolopyrimidine was further validated in embryonic and adult zebrafish [ 124 ]. As the vascular system is the first functional organ to develop in the mammalian embryo, and the disruption of the vascular system has been correlated with fetal loss, human malformations, and cognitive impairment, these generated platform is important for the identification of compounds with developmental toxicity.…”