Targeted Inhibition of Leucine-Rich Repeat and Immunoglobulin Domain-Containing Protein 1 in Transplanted Neural Stem Cells Promotes Neuronal Differentiation and Functional Recovery in Rats Subjected to Spinal Cord Injury.
OBJECTIVE
Leucine-rich repeat and immunoglobulin domain-containing protein (LINGO)-1 is expressed in neural stem cells, and its neutralization results in sustained neuronal immaturity. Thus, targeted inhibition of LINGO-1 via RNA interference may enhance transplanted neural stem cell survival and neuronal differentiation in vivo. Furthermore, LINGO-1 RNA interference in neural stem cells represents a potential therapeutic strategy for spinal cord injury.


METHODS
Department of Spine Surgery, First Affiliated Hospital of Sun Yat-sen University.


METHODS
Translational Medicine Center Research Laboratory, First Affiliated Hospital of Sun Yat-sen University.


METHODS
Female Sprague-Dawley rats.


METHODS
The animals were divided into three groups that underwent laminectomy and complete spinal cord transection accompanied by transplantation of control-RNA interference-treated or LINGO-1-RNA interference-treated neural stem cells at the injured site in vivo. In vitro, neural stem cells were divided into four groups for the following treatments: control, control RNA interference lentivirus, LINGO-1 RNA interference lentivirus and LINGO-1 complementary DNA lentivirusand the Key Projects of the Natural Science Foundation of Guangdong Province (No. S2013020012818).


RESULTS
Neural stem cells in each treatment group were examined for cell survival and neuronal differentiation in vitro and in vivo via immunofluorescence and Western blot analysis. Axonal regeneration and tissue repair were assessed via retrograde tracing using Fluorogold, electron microscopy, hematoxylin-eosin staining and MRI. Rats were also examined for functional recovery based on the measurement of spinal cord-evoked potentials and the Basso-Beattie-Bresnahan score. LINGO-1-RNA interference-treated neural stem cell transplantation increased tissue repair and functional recovery of the injured spinal cord in rats. Similarly, LINGO-1 RNA interference increased neural stem cell survival and neuronal differentiation in vitro. The mechanism underlying the effect of LINGO-1 RNA interference on the injured rat spinal cord may be that the significant inhibition of LINGO-1 expression in neural stem cells inactivated the RhoA and Notch signaling pathways, which act downstream of LINGO-1.


CONCLUSIONS
Our findings indicate that transplantation of LINGO-1-RNA interference-treated neural stem cells facilitates functional recovery after spinal cord injury and represents a promising potential strategy for the repair of spinal cord injury.
1Department of Spine Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.2Department of Orthopedics, Foshan Hospital of Traditional Chinese Medicine, Foshan, China.3Department of Translational Medicine Center Research Laboratory, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.4Department of Medicine-Cardiology and Department of Molecular Pharmacology and Biological Chemistry, Northwestern University Feinberg School of Medicine, Chicago, IL.5Department of Hepatopancreatobiliary Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.
Ningning
N
Chen
Jing-Sheng
JS
Cen
Jingnan
J
Wang
Gangjian
G
Qin
Lingli
L
Long
Le
L
Wang
Fuxin
F
Wei
Qingfeng
Q
Xiang
David Y B
DY
Deng
Yong
Y
Wan
D000818:Animals; D005260:Female; D005822:Genetic Vectors; D007278:Injections, Spinal; D007796:Laminectomy; D016086:Lentivirus / Q000235:genetics; D008565:Membrane Proteins / Q000037:antagonists & inhibitors*; D009416:Nerve Regeneration; D009419:Nerve Tissue Proteins / Q000008:administration & dosage* / Q000037:antagonists & inhibitors / Q000235:genetics; D058953:Neural Stem Cells / Q000637:transplantation*; D034622:RNA Interference; D051381:Rats; D017207:Rats, Sprague-Dawley; D020127:Recovery of Function / Q000502:physiology; D013116:Spinal Cord / Q000502:physiology; D013119:Spinal Cord Injuries / Q000628:therapy*; D033581:Stem Cell Transplantation / Q000379:methods*
C525164:LINGO1 protein, rat; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins