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.

A hybrid microfluidic system for regulation of neural differentiation in induced pluripotent stem cells

“…Stem cells play a pivotal role in the human body for tissue regeneration and they are now used as an integral part of modern clinical treatment . Over the past decades, numerous attempts are devoted toward neural differentiation of stem cells . Various sources of stem cells such as neural stem cells (NSCs) or neural progenitor cells (NPCs) have been studied for treatment of SCI and varying levels of improvement in animal models have been demonstrated .…”

Nucleolipids of Canonical Purine ß‐d‐Ribo‐Nucleosides: Synthesis and Cytostatic/Cytotoxic Activities Toward Human and Rat Glioblastoma Cells

“…In a series of precedent publications we and others have demonstrated that the cancerostatic/cancerotoxic activity of pyrimidine ß‐ d ‐ribonucleoside antimetabolites such as 5‐fluorouridine and 6‐azauridine towards different human tumor cell lines as well as neurobiological and antiviral activities can be significantly improved by lipophilization. Also, regular, canonical pyrimidine ß ‐ d ‐ribonucleosides such as uridine and 5‐methyluridine acquire a surprisingly high antitumor in vitro activity upon covalent hydrophobization .…”