Development and Use of an Embryonic Stem Cell-Derived Spinal Motoneuron Assay for the Evaluation of Potential Treatments Against Alphaviruses

Abstract

Venezuelan (VEEV), eastern (EEEV), and western (WEEV) equine encephalitis viruses are positive-stranded RNA viruses transmitted by mosquitos and cause disease in horses and humans. Current methods to evaluate potential therapeutics in vitro use immortalized cell lines, but their tumorigenic properties result in genetic and physiologic differences from the primary cells they mimic. Primary cells while physiologically relevant, are difficult to produce in large quantities. Stem cells can be produced in larger quantities than primary cells and are more physiologically relevant than immortalized cell lines. For this work, embryonic stem (ES) cell-derived motoneurons resembling native motor neurons both physiologically and morphologically were utilized to develop a screening method to allow for better down selection of compounds with anti-alphavirus efficacy prior to in vivo experiments Several compounds were screened in the VEEV ES cell-derived motoneurons assay and one compound was down selected and evaluated for in vivo efficacy.