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 (Academic Journal)peer-review

24 Citations (Scopus)

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
Pages (from-to)1235-44
Number of pages10
JournalCytotherapy
Volume14
Issue number10
DOIs
Publication statusPublished - Nov 2012

Keywords

  • Animals
  • Antigens/genetics
  • CD11b Antigen/metabolism
  • Cellular Microenvironment
  • Chondroitin Sulfate Proteoglycans/metabolism
  • Cicatrix/pathology
  • Female
  • Gene Transfer Techniques
  • Genetic Therapy
  • Genetic Vectors/genetics
  • Glial Fibrillary Acidic Protein/metabolism
  • Immunohistochemistry
  • Lentivirus/genetics
  • Locomotion
  • Neurotrophin 3/genetics
  • Proteoglycans/genetics
  • RNA, Small Interfering/administration & dosage
  • Rats
  • Rats, Sprague-Dawley
  • Recovery of Function
  • Spinal Cord Injuries/pathology
  • Staining and Labeling
  • Tubulin/metabolism

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