Identification of Nucleoside Analogs as Inducers of Neuronal Differentiation in a Human Reporter Cell Line and Adult Stem Cells

• Published in: Chemical biology & drug design 2015-08, Vol.86 (2), p.129-143
• Main Authors: Raasch, Katharina, Malecki, Edith, Siemann, Maria, Martinez, Malayko M., Heinisch, Jürgen J., Müller, Janine, Bakota, Lidia, Kaltschmidt, Christian, Kaltschmidt, Barbara, Rosemeyer, Helmut, Brandt, Roland
• Format: Article
• Language: English
• Subjects: Adult; adult human stem cell; Adult Stem Cells - cytology; Adult Stem Cells - drug effects; Cell Differentiation - drug effects; Cell Line; cladribine; Drug Evaluation, Preclinical - methods; Embryonal Carcinoma Stem Cells; human model neuron; Humans; neuronal differentiation; Neurons - cytology; Neurons - drug effects; nucleoside analog; Nucleosides - chemical synthesis; Nucleosides - chemistry; Nucleosides - pharmacology
• ISSN: 1747-0277; 1747-0285
• DOI: 10.1111/cbdd.12488

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. Nucleoside analogues (NSAs) were among the first chemotherapeutic agents and could also be useful for manipulation of cell fate. For screening of NSAs that influence neuronal differentiation, we developed a novel phenotypic assay based on a human neuron‐committed teratocarcinoma cell line (NT2) as a model for neuronal progenitors. In a small scale screen we tested a panel of structurally related NSAs and report that 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.