title: 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.
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal.
authors.forename: Helena
authors.initials: H
authors.lastname: Vazão
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal.
authors.forename: Susana
authors.initials: S
authors.lastname: Rosa
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal.|MIRA Institute for Biomedical Engineering and Technical Medicine, University Twente, Enschede, 7500AE, The Netherlands.
authors.forename: Tânia
authors.initials: T
authors.lastname: Barata
authors.affiliation: Department of Cardiovascular Development and Repair, Centro Nacional de Investigaciones Cardiovasculares Carlos III, 28029 Madrid, Spain.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Costa
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal.
authors.forename: Patrícia R
authors.initials: PR
authors.lastname: Pitrez
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal.
authors.forename: Inês
authors.initials: I
authors.lastname: Honório
authors.affiliation: Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands.
authors.forename: Margreet R
authors.initials: MR
authors.lastname: de Vries
authors.affiliation: Black Family Stem Cell Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029.
authors.forename: Dimitri
authors.initials: D
authors.lastname: Papatsenko
authors.affiliation: Department of Cardiovascular Development and Repair, Centro Nacional de Investigaciones Cardiovasculares Carlos III, 28029 Madrid, Spain.
authors.forename: Rui
authors.initials: R
authors.lastname: Benedito
authors.affiliation: MIRA Institute for Biomedical Engineering and Technical Medicine, University Twente, Enschede, 7500AE, The Netherlands.
authors.forename: Daniel
authors.initials: D
authors.lastname: Saris
authors.affiliation: Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115.|Harvard-Massachusetts Institute of Technology Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139.|Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115.|Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul 130-701, Republic of Korea.|Department of Physics, King Abdulaziz University, Jeddah 21569, Saudi Arabia.
authors.forename: Ali
authors.initials: A
authors.lastname: Khademhosseini
authors.affiliation: Einthoven Laboratory for Experimental Vascular Medicine, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands.
authors.forename: Paul H A
authors.initials: PH
authors.lastname: Quax
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal.
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Pereira
authors.affiliation: Institute of Anatomy, University of Bern, 3012 Bern, Switzerland.
authors.forename: Nadia
authors.initials: N
authors.lastname: Mercader
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal.|MIRA Institute for Biomedical Engineering and Technical Medicine, University Twente, Enschede, 7500AE, The Netherlands.
authors.forename: Hugo
authors.identifier: 0000-0002-4574-7648
authors.initials: H
authors.lastname: Fernandes
authors.affiliation: Center for Neurosciences and Cell Biology, University of Coimbra, 3000 Coimbra, Portugal; lino@uc-biotech.pt.
authors.forename: Lino
authors.initials: L
authors.lastname: Ferreira
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D054852:Small Molecule Libraries / Q000494:pharmacology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000037:antagonists & inhibitors; D015027:Zebrafish / Q000196:embryology / Q000254:growth & development* / Q000378:metabolism
chemical_list: D054852:Small Molecule Libraries; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: embryonic endothelial markers; endothelial cells; high-throughput screening; pluripotent stem cells; vascular toxicity