Transduced Schwann cells promote axon growth and myelination after spinal cord injury.
We sought to directly compare growth and myelination of local and supraspinal axons by implanting into the injured spinal cord Schwann cells (SCs) transduced ex vivo with adenoviral (AdV) or lentiviral (LV) vectors encoding a bifunctional neurotrophin molecule (D15A). D15A mimics actions of both neurotrophin-3 and brain-derived neurotrophic factor. Transduced SCs were injected into the injury center 1 week after a moderate thoracic (T8) adult rat spinal cord contusion. D15A expression and bioactivity in vitro; D15A levels in vivo; and graft volume, SC number, implant axon number and cortico-, reticulo-, raphe-, coerulo-spinal and sensory axon growth were determined for both types of vectors employed to transduce SCs. ELISAs revealed that D15A-secreting SC implants contained significantly higher levels of neurotrophin than non-transduced SC and AdV/GFP and LV/GFP SC controls early after implantation. At 6 weeks post-implantation, D15A-secreting SC grafts exhibited 5-fold increases in graft volume, SC number and myelinated axon counts and a 3-fold increase in myelinated to unmyelinated (ensheathed) axon ratios. The total number of axons within grafts of LV/GFP/D15A SCs was estimated to be over 70,000. Also 5-HT, DbetaH, and CGRP axon length was increased up to 5-fold within D15A grafts. In sum, despite qualitative differences using the two vectors, increased neurotrophin secretion by the implanted D15A SCs led to the presence of a significantly increased number of axons in the contusion site. These results demonstrate the therapeutic potential for utilizing neurotrophin-transduced SCs to repair the injured spinal cord.
Golden
Kevin L
KL
The Miami Project to Cure Paralysis and the Neuroscience Program, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33101, USA.
Pearse
Damien D
DD
Blits
Bas
B
Garg
Maneesh S
MS
Oudega
Martin
M
Wood
Patrick M
PM
Bunge
Mary Bartlett
MB
D000704:Analysis of Variance; D000818:Animals; D001369:Axons / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D001522:Behavior, Animal; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism; D017690:Cell Transplantation / Q000379:methods; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D009414:Nerve Growth Factors / Q000378:metabolism*; D009416:Nerve Regeneration / Q000502:physiology; D051381:Rats; D011916:Rats, Inbred F344; D012583:Schwann Cells / Q000502:physiology*; D012584:Sciatic Nerve / Q000166:cytology; D012701:Serotonin / Q000378:metabolism; D013119:Spinal Cord Injuries / Q000601:surgery*; D014161:Transduction, Genetic / Q000379:methods*
D009414:Nerve Growth Factors; D049452:Green Fluorescent Proteins; D012701:Serotonin; D004299:Dopamine beta-Hydroxylase; D015740:Calcitonin Gene-Related Peptide
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