title: Impairment of human neural crest cell migration by prolonged exposure to interferon-beta.
abstract: Human cell-based toxicological assays have been used successfully to detect known toxicants, and to distinguish them from negative controls. However, there is at present little experience on how to deal with hits from screens of compounds with yet unknown hazard. As a case study to this issue, we characterized human interferon-beta (IFNβ) as potential developmental toxicant affecting neural crest cells (NCC). The protein was identified as a hit during a screen of clinically used drugs in the 'migration inhibition of neural crest' (MINC) assay. Concentration-response studies in the MINC combined with immunocytochemistry and mRNA quantification of cellular markers showed that IFNβ inhibited NCC migration at concentrations as low as 20 pM. The effective concentrations found here correspond to levels found in human plasma, and they were neither cytostatic nor cytotoxic nor did they did they affect the differentiation state and overall phenotype of NCC. Data from two other migration assays confirmed that picomolar concentration of IFNβ reduced the motility of NCC, while other interferons were less potent. The activation of JAK kinase by IFNβ, as suggested by bioinformatics analysis of the transcriptome changes, was confirmed by biochemical methods. The degree and duration of pathway activation correlated with the extent of migration inhibition, and pharmacological block of this signaling pathway before, or up to 6 h after exposure to the cytokine prevented the effects of IFNβ on migration. Thus, the reduction of vital functions of human NCC is a hitherto unknown potential hazard of endogenous or pharmacologically applied interferons.
authors.affiliation: Department of In Vitro Toxicology and Biomedicine, University of Konstanz, Box 657, 78457, Konstanz, Germany. giorgia.pallocca@uni-konstanz.de.
authors.forename: Giorgia
authors.identifier: http://orcid.org/0000-0002-7358-1788
authors.initials: G
authors.lastname: Pallocca
authors.affiliation: Department of In Vitro Toxicology and Biomedicine, University of Konstanz, Box 657, 78457, Konstanz, Germany.
authors.forename: Johanna
authors.initials: J
authors.lastname: Nyffeler
authors.affiliation: Department of In Vitro Toxicology and Biomedicine, University of Konstanz, Box 657, 78457, Konstanz, Germany.
authors.forename: Xenia
authors.initials: X
authors.lastname: Dolde
authors.affiliation: Department of Statistics, TU Dortmund, 44139, Dortmund, Germany.
authors.forename: Marianna
authors.initials: M
authors.lastname: Grinberg
authors.affiliation: Department of Physiology, University of Innsbruck, 6020, Innsbruck, Austria.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Gstraunthaler
authors.affiliation: Department of In Vitro Toxicology and Biomedicine, University of Konstanz, Box 657, 78457, Konstanz, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Waldmann
authors.affiliation: Department of Statistics, TU Dortmund, 44139, Dortmund, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Rahnenführer
authors.affiliation: Center of Physiology and Pathophysiology, Institute of Neurophysiology, University of Cologne, 50931, Cologne, Germany.
authors.forename: Agapios
authors.initials: A
authors.lastname: Sachinidis
authors.affiliation: Department of In Vitro Toxicology and Biomedicine, University of Konstanz, Box 657, 78457, Konstanz, Germany.
authors.forename: Marcel
authors.initials: M
authors.lastname: Leist
mesh_terms: D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D020869:Gene Expression Profiling; D006801:Humans; D016899:Interferon-beta / Q000633:toxicity*; D053612:Janus Kinases / Q000378:metabolism; D009432:Neural Crest / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D050791:STAT Transcription Factors / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D050791:STAT Transcription Factors; D016899:Interferon-beta; D053612:Janus Kinases
keywords: Cell migration; Developmental toxicity; Interferons; JAK-STAT pathway; Neural crest