Canine olfactory ensheathing cells from the olfactory mucosa can be engineered to produce active chondroitinase ABC

Darren Carwardine, Liang-Fong Wong, James Fawcett, Elizabeth Muir, Nicolas Granger

Research output: Contribution to journalArticle (Academic Journal)peer-review

7 Citations (Scopus)
319 Downloads (Pure)

Abstract

A multitude of factors must be overcome following spinal cord injury (SCI) in order to achieve clinical improvement in patients. It is thought that by combining promising therapies these diverse factors could be combatted with the aim of producing an overall improvement in function. Chondroitin sulphate proteoglycans (CSPGs) present in the glial scar that forms following SCI present a significant block to axon regeneration. Digestion of CSPGs by chondroitinase ABC (ChABC) leads to axon regeneration, neuronal plasticity and functional improvement in preclinical models of SCI. However, the enzyme activity decays at body temperature within 24–72 h, limiting the translational potential of ChABC as a therapy. Olfactory ensheathing cells (OECs) have shown huge promise as a cell transplant therapy in SCI. Their beneficial effects have been demonstrated in multiple small animal SCI models as well as in naturally occurring SCI in canine patients. In the present study, we have genetically modified canine OECs from the mucosa to constitutively produce enzymatically active ChABC. We have developed a lentiviral vector that can deliver a mammalian modified version of the ChABC gene to mammalian cells, including OECs. Enzyme production was quantified using the Morgan-Elson assay that detects the breakdown products of CSPG digestion in cell supernatants. We confirmed our findings by immunolabelling cell supernatant samples using Western blotting. OECs normal cell function was unaffected by genetic modification as demonstrated by normal microscopic morphology and the presence of the low affinity neurotrophin receptor (p75NGF) following viral transduction. We have developed the means to allow production of active ChABC in combination with a promising cell transplant therapy for SCI repair.
Original languageEnglish
Pages (from-to)311-318
Number of pages8
JournalJournal of the Neurological Sciences
Volume367
Early online date5 Jun 2016
DOIs
Publication statusPublished - 15 Aug 2016

Keywords

  • Chondroitinase ABC
  • Chondroitin sulphate proteoglycan
  • Olfactory ensheathing cells
  • Olfactory mucosa
  • Spinal cord injury
  • Gene therapy

Fingerprint Dive into the research topics of 'Canine olfactory ensheathing cells from the olfactory mucosa can be engineered to produce active chondroitinase ABC'. Together they form a unique fingerprint.

Cite this