title: Distinct gene expression responses of two anticonvulsant drugs in a novel human embryonic stem cell based neural differentiation assay protocol.
abstract: Hazard assessment of chemicals and pharmaceuticals is increasingly gaining from knowledge about molecular mechanisms of toxic action acquired in dedicated in vitro assays. We have developed an efficient human embryonic stem cell neural differentiation test (hESTn) that allows the study of the molecular interaction of compounds with the neural differentiation process. Within the 11-day differentiation protocol of the assay, embryonic stem cells lost their pluripotency, evidenced by the reduced expression of stem cell markers Pou5F1 and Nanog. Moreover, stem cells differentiated into neural cells, with morphologically visible neural structures together with increased expression of neural differentiation-related genes such as βIII-tubulin, Map2, Neurogin1, Mapt and Reelin. Valproic acid (VPA) and carbamazepine (CBZ) exposure during hESTn differentiation led to concentration-dependent reduced expression of βIII-tubulin, Neurogin1 and Reelin. In parallel VPA caused an increased gene expression of Map2 and Mapt which is possibly related to the neural protective effect of VPA. These findings illustrate the added value of gene expression analysis for detecting compound specific effects in hESTn. Our findings were in line with and could explain effects observed in animal studies. This study demonstrates the potential of this assay protocol for mechanistic analysis of specific compound-induced inhibition of human neural cell differentiation.
authors.affiliation: Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands; Institute for Risk Assessment Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, The Netherlands. Electronic address: sjors.schulpen@rivm.nl.
authors.forename: Sjors H W
authors.initials: SH
authors.lastname: Schulpen
authors.affiliation: Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands; Institute for Risk Assessment Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, The Netherlands.
authors.forename: Esther
authors.initials: E
authors.lastname: de Jong
authors.affiliation: Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands.
authors.forename: Liset J J
authors.initials: LJ
authors.lastname: de la Fonteyne
authors.affiliation: Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands.
authors.forename: Arja
authors.initials: A
authors.lastname: de Klerk
authors.affiliation: Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands; Institute for Risk Assessment Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, The Netherlands.
authors.forename: Aldert H
authors.initials: AH
authors.lastname: Piersma
mesh_terms: D000818:Animals; D000927:Anticonvulsants / Q000494:pharmacology*; D002220:Carbamazepine / Q000633:toxicity; D002454:Cell Differentiation / Q000187:drug effects*; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D005347:Fibroblasts; D015870:Gene Expression / Q000187:drug effects*; D006801:Humans; D051379:Mice; D009474:Neurons / Q000187:drug effects*; D012313:RNA / Q000096:biosynthesis; D000091042:Reelin Protein; D014635:Valproic Acid / Q000633:toxicity
chemical_list: D000927:Anticonvulsants; D000091042:Reelin Protein; D002220:Carbamazepine; D014635:Valproic Acid; D012313:RNA; C000717127:RELN protein, human; C000717128:Reln protein, mouse
keywords: Carbamazepine; Human embryonic stem cells; Neural differentiation; Neurodevelopmental toxicity; Valproic acid