title: Identification of Nucleoside Analogs as Inducers of Neuronal Differentiation in a Human Reporter Cell Line and Adult Stem Cells.
abstract: Nucleoside analogs (NSAs) were among the first chemotherapeutic agents and could also be useful for the manipulation of cell fate. To investigate the potential of NSAs for the induction of neuronal differentiation, we developed a novel phenotypic assay based on a human neuron-committed teratocarcinoma cell line (NT2) as a model for neuronal progenitors and constructed a NT2-based reporter cell line that expressed eGFP under the control of a neuron-specific promoter. We tested 38 structurally related NSAs and determined their activity to induce neuronal differentiation by immunocytochemistry of neuronal marker proteins, live cell imaging, fluorometric detection and immunoblot analysis. We identified twelve NSAs, which induced neuronal differentiation to different extents. NSAs with highest activity carried a halogen substituent at their pyrimidine nucleobase and an unmodified or 2'-O-methyl substituted 2-deoxy-β-D-ribofuranosyl residue as glyconic moiety. Cladribine, a purine nucleoside with similar structural features and in use to treat leukemia and multiple sclerosis, induced also differentiation of adult human neural crest-derived stem cells. Our results suggest that NSAs could be useful for the manipulation of neuronal cell fate in cell replacement therapy or treatment of neurodegenerative disorders. The data on the structure and function relationship will help to design compounds with increased activity and low toxicity.
authors.affiliation: Department of Neurobiology, University of Osnabrück, Barbarastrasse 11, 49076, Osnabrück, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Raasch
authors.affiliation: Institute of Chemistry of New Materials, University of Osnabrück, Barbarastrasse 7, 49076, Osnabrück, Germany.
authors.forename: Edith
authors.initials: E
authors.lastname: Malecki
authors.affiliation: Department of Neurobiology, University of Osnabrück, Barbarastrasse 11, 49076, Osnabrück, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Siemann
authors.affiliation: Department of Neurobiology, University of Osnabrück, Barbarastrasse 11, 49076, Osnabrück, Germany.
authors.forename: Malayko M
authors.initials: MM
authors.lastname: Martinez
authors.affiliation: Department of Genetics, University of Osnabrück, Barbarastrasse 11, 49076, Osnabrück, Germany.
authors.forename: Jürgen J
authors.initials: JJ
authors.lastname: Heinisch
authors.affiliation: Department of Molecular Neurobiology, University of Bielefeld, Universitätsstrasse 25, 33615, Bielefeld, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Müller
authors.affiliation: Department of Neurobiology, University of Osnabrück, Barbarastrasse 11, 49076, Osnabrück, Germany.
authors.forename: Lidia
authors.initials: L
authors.lastname: Bakota
authors.affiliation: Department of Molecular Neurobiology, University of Bielefeld, Universitätsstrasse 25, 33615, Bielefeld, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Kaltschmidt
authors.affiliation: Department of Molecular Neurobiology, University of Bielefeld, Universitätsstrasse 25, 33615, Bielefeld, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Kaltschmidt
authors.affiliation: Institute of Chemistry of New Materials, University of Osnabrück, Barbarastrasse 7, 49076, Osnabrück, Germany.
authors.forename: Helmut
authors.initials: H
authors.lastname: Rosemeyer
authors.affiliation: Department of Neurobiology, University of Osnabrück, Barbarastrasse 11, 49076, Osnabrück, Germany.
authors.forename: Roland
authors.initials: R
authors.lastname: Brandt
mesh_terms: D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D004353:Drug Evaluation, Preclinical / Q000379:methods; D054278:Embryonal Carcinoma Stem Cells; D006801:Humans; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D009705:Nucleosides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*
chemical_list: D009705:Nucleosides
keywords: adult human stem cell; cladribine; human model neuron; neuronal differentiation; nucleoside analog