Activation of Neurogenesis in Multipotent Stem Cells CulturedIn Vitroand in the Spinal Cord Tissue After Severe Injury by Inhibition of Glycogen Synthase Kinase-3

Autores de CIPF

• 

  Francisco Rodríguez Jiménez

  Autor

• Eleonora Clemente

  Autor

• Raquel Román Parrilla

  Autor

• Ana Artero Castro

  Autor

• Santos Fustero Lardies

  Autor

• 

  M Victoria Moreno Manzano

  Autor

• 

  Slaven Erceg Vukicevic

  Autor

Participantes ajenos a CIPF

• Vilches, A
• Perez-Arago, MA
• Leal, J
• Jendelova, P
• Stojkovic, M

Grupos de Investigación

• Laboratorio de Regeneración Tisular y Neuronal

  

  Jefe/a de Grupo

  M Victoria Moreno Manzano

• Laboratorio Terapias con Células Madre en Enfermedades Neurodegenerativas

  

  Jefe/a de Grupo

  Slaven Erceg Vukicevic

Abstract

The inhibition of glycogen synthase kinase-3 (GSK-3) can induce neurogenesis, and the associated activation of Wnt/beta-catenin signaling via GSK-3 inhibition may represent a means to promote motor function recovery following spinal cord injury (SCI) via increased astrocyte migration, reduced astrocyte apoptosis, and enhanced axonal growth. Herein, we assessed the effects of GSK-3 inhibitionin vitroon the neurogenesis of ependymal stem/progenitor cells (epSPCs) resident in the mouse spinal cord and of human embryonic stem cell-derived neural progenitors (hESC-NPs) and human-induced pluripotent stem cell-derived neural progenitors (hiPSC-NPs) andin vivoon spinal cord tissue regeneration and motor activity after SCI. We report that the treatment of epSPCs and human pluripotent stem cell-derived neural progenitors (hPSC-NPs) with the GSK-3 inhibitor Ro3303544 activates beta-catenin signaling and increases the expression of the bIII-tubulin neuronal marker; furthermore, the differentiation of Ro3303544-treated cells prompted an increase in the number of terminally differentiated neurons. Administration of a water-soluble, bioavailable form of this GSK-3 inhibitor (Ro3303544-Cl) in a severe SCI mouse model revealed the increased expression of bIII-tubulin in the injury epicenter. Treatment with Ro3303544-Cl increased survival of mature neuron types from the propriospinal tract (vGlut1, Parv) and raphe tract (5-HT), protein kinase C gamma-positive neurons, and GABAergic interneurons (GAD65/67) above the injury epicenter. Moreover, we observed higher numbers of newly born BrdU/DCX-positive neurons in Ro3303544-Cl-treated animal tissues, a reduced area delimited by astrocyte scar borders, and improved motor function. Based on this study, we believe that treating animals with epSPCs or hPSC-NPs in combination with Ro3303544-Cl deserves further investigation towards the development of a possible therapeutic strategy for SCI.

Keywords

• Spinal cord injury; stem cells; neurogenesis; axonal growth; GSK3 inhibition