title: Development of a pluripotent stem cell derived neuronal model to identify chemically induced pathway perturbations in relation to neurotoxicity: effects of CREB pathway inhibition.
abstract: According to the advocated paradigm shift in toxicology, acquisition of knowledge on the mechanisms underlying the toxicity of chemicals, such as perturbations of biological pathways, is of primary interest. Pluripotent stem cells (PSCs), such as human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), offer a unique opportunity to derive physiologically relevant human cell types to measure molecular and cellular effects of such pathway modulations. Here we compared the neuronal differentiation propensity of hESCs and hiPSCs with the aim to develop novel hiPSC-based tools for measuring pathway perturbation in relation to molecular and cellular effects in vitro. Among other fundamental pathways, also, the cAMP responsive element binding protein (CREB) pathway was activated in our neuronal models and gave us the opportunity to study time-dependent effects elicited by chemical perturbations of the CREB pathway in relation to cellular effects. We show that the inhibition of the CREB pathway, using 2-naphthol-AS-E-phosphate (KG-501), induced an inhibition of neurite outgrowth and synaptogenesis, as well as a decrease of MAP2(+) neuronal cells. These data indicate that a CREB pathway inhibition can be related to molecular and cellular effects that may be relevant for neurotoxicity testing, and, thus, qualify the use of our hiPSC-derived neuronal model for studying chemical-induced neurotoxicity resulting from pathway perturbations.
authors.affiliation: Institute for Health and Consumer Protection (IHCP), JRC, Ispra, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Pistollato
authors.affiliation: Institute for Health and Consumer Protection (IHCP), JRC, Ispra, Italy.
authors.forename: Jochem
authors.initials: J
authors.lastname: Louisse
authors.affiliation: Institute for Health and Consumer Protection (IHCP), JRC, Ispra, Italy.
authors.forename: Bibiana
authors.initials: B
authors.lastname: Scelfo
authors.affiliation: Institute for Health and Consumer Protection (IHCP), JRC, Ispra, Italy.
authors.forename: Milena
authors.initials: M
authors.lastname: Mennecozzi
authors.affiliation: Oncohematology Laboratory, Department of Woman and Child Health, University of Padova, Padova, Italy.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Accordi
authors.affiliation: Oncohematology Laboratory, Department of Woman and Child Health, University of Padova, Padova, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Basso
authors.affiliation: Center of Physiology and Pathophysiology, Institute of Neurophysiology, University of Cologne, Cologne, Germany.
authors.forename: John Antonydas
authors.initials: JA
authors.lastname: Gaspar
authors.affiliation: Institute for Health and Consumer Protection (IHCP), JRC, Ispra, Italy.
authors.forename: Dimitra
authors.initials: D
authors.lastname: Zagoura
authors.affiliation: Institute for Health and Consumer Protection (IHCP), JRC, Ispra, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Barilari
authors.affiliation: Institute for Health and Consumer Protection (IHCP), JRC, Ispra, Italy.
authors.forename: Taina
authors.initials: T
authors.lastname: Palosaari
authors.affiliation: Center of Physiology and Pathophysiology, Institute of Neurophysiology, University of Cologne, Cologne, Germany.
authors.forename: Agapios
authors.initials: A
authors.lastname: Sachinidis
authors.affiliation: Institute for Health and Consumer Protection (IHCP), JRC, Ispra, Italy. Electronic address: susanne.bremer@jrc.ec.europa.eu.
authors.forename: Susanne
authors.initials: S
authors.lastname: Bremer-Hoffmann
mesh_terms: D002454:Cell Differentiation; D017362:Cyclic AMP Response Element-Binding Protein / Q000037:antagonists & inhibitors / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000166:cytology; D020869:Gene Expression Profiling; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D009284:Naphthols / Q000494:pharmacology; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D010755:Organophosphates / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects*; D013569:Synapses / Q000187:drug effects
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D009284:Naphthols; D010755:Organophosphates; C495860:naphthol AS-E phosphate
keywords: CREB pathway; Embryonic stem cells; Induced pluripotent stem cells; KG-501; Neuronal derivatives