Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord

Eleanor M. Donnelly, Nicolas N. Madigan, Gemma E. Rooney, Andrew Knight, Bingkun Chen, Bret Ball, Lisa Kinnavane, Yolanda Garcia, Peter Dockery, John Fraher, Padraig M. Strappe, Anthony J. Windebank, Timothy O'Brien, Siobhan S. McMahon

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

Background aims. In this study we investigated the effect of neurotrophin-3 (NT-3) and knockdown of NG2, one of the main inhibitory chondroitin sulfate proteoglycans (CSPG), in the glial scar following spinal cord injury (SCI). Methods. Short hairpin (sh) RNA were designed to target NG2 and were cloned into a lentiviral vector (LV). A LV was also constructed containing NT-3. LV expressing NT-3, shRNA to NG2 or combinations of both vectors were injected directly into contused rat spinal cords 1 week post-injury. Six weeks post-injection of LV, spinal cords were examined by histology for changes in scar size and by immunohistochemistry for changes in expression of CSPG, NT-3, astrocytes, neurons and microglia/macrophages. Motor function was assessed using the Basso, Beattie and Bresnahan (BBB) locomotor scale. Results. Animals that received the combination treatment of LV shNG2 and LV NT-3 showed reduced scar size. These animals also showed an increase in levels of neurons and NG2, a decrease in levels of astrocytes and a significant functional recovery as assessed using the BBB locomotor scale at 2 weeks post-treatment. Conclusions. The improvement in locomotor recovery and decrease in scar size shows the potential of this gene therapy approach as a therapeutic treatment for SCI.

Original languageEnglish (US)
Pages (from-to)1235-1244
Number of pages10
JournalCytotherapy
Volume14
Issue number10
DOIs
StatePublished - Nov 2012

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Keywords

  • NG2
  • astrocytes
  • axon regeneration
  • chondroitin sulfate proteoglycans
  • glial scar
  • microglia
  • neurotrophin-3
  • spinal cord injury

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Oncology
  • Genetics(clinical)
  • Cell Biology
  • Cancer Research
  • Transplantation

Cite this

Donnelly, E. M., Madigan, N. N., Rooney, G. E., Knight, A., Chen, B., Ball, B., Kinnavane, L., Garcia, Y., Dockery, P., Fraher, J., Strappe, P. M., Windebank, A. J., O'Brien, T., & McMahon, S. S. (2012). Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord. Cytotherapy, 14(10), 1235-1244. https://doi.org/10.3109/14653249.2012.714865