Highlights
- •Olfactory ensheathing cells from the mucosa can be genetically modified to secrete chondroitinase ABC.
- •Lentiviral transduction of olfactory ensheathing cells did not change their morphology or cell marker expression (p75NGF).
- •Chondroitinase ABC production was confirmed using the Morgan-Elson assay and Western blot.
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.
Keywords
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Article info
Publication history
Published online: June 05, 2016
Accepted:
June 3,
2016
Received in revised form:
June 1,
2016
Received:
February 25,
2016
Identification
Copyright
© 2016 Published by Elsevier B.V.
