Skip to main content

Research Repository

Advanced Search

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

Carwardine, D; Wong, L F; Fawcett, J W; Muir, E M; Granger, N

Authors

D Carwardine

L F Wong

J W Fawcett

E M Muir

N Granger



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.

Citation

Carwardine, D., Wong, L. F., Fawcett, J. W., Muir, E. M., & Granger, N. (2016). Canine olfactory ensheathing cells from the olfactory mucosa can be engineered to produce active chondroitinase ABC. Journal of the Neurological Sciences, 367, 311-8. https://doi.org/10.1016/j.jns.2016.06.011

Journal Article Type Article
Acceptance Date Jun 3, 2016
Publication Date Aug 15, 2016
Deposit Date Aug 21, 2018
Journal Journal of the Neurological Sciences
Print ISSN 0022-510X
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 367
Pages 311-8
DOI https://doi.org/10.1016/j.jns.2016.06.011
Public URL https://rvc-repository.worktribe.com/output/1395843