Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord

Eleanor M. Donnelly; Nicolas N. Madigan; Gemma E. Rooney; Andrew Knight; Bingkun Chen; Bret Ball; Lisa Kinnavane; Yolanda Garcia; Peter Dockery; John Fraher; Padraig M. Strappe; Anthony J. Windebank; Timothy O'Brien; Siobhan S. McMahon

doi:10.3109/14653249.2012.714865

Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord

Eleanor M. Donnelly, Nicolas N. Madigan, Gemma E. Rooney, Andrew Knight, Bingkun Chen, Bret Ball, Lisa Kinnavane, Yolanda Garcia, Peter Dockery, John Fraher, Padraig M. Strappe, Anthony J. Windebank, Timothy O'Brien, Siobhan S. McMahon

Neurology

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

Background aims. In this study we investigated the effect of neurotrophin-3 (NT-3) and knockdown of NG2, one of the main inhibitory chondroitin sulfate proteoglycans (CSPG), in the glial scar following spinal cord injury (SCI). Methods. Short hairpin (sh) RNA were designed to target NG2 and were cloned into a lentiviral vector (LV). A LV was also constructed containing NT-3. LV expressing NT-3, shRNA to NG2 or combinations of both vectors were injected directly into contused rat spinal cords 1 week post-injury. Six weeks post-injection of LV, spinal cords were examined by histology for changes in scar size and by immunohistochemistry for changes in expression of CSPG, NT-3, astrocytes, neurons and microglia/macrophages. Motor function was assessed using the Basso, Beattie and Bresnahan (BBB) locomotor scale. Results. Animals that received the combination treatment of LV shNG2 and LV NT-3 showed reduced scar size. These animals also showed an increase in levels of neurons and NG2, a decrease in levels of astrocytes and a significant functional recovery as assessed using the BBB locomotor scale at 2 weeks post-treatment. Conclusions. The improvement in locomotor recovery and decrease in scar size shows the potential of this gene therapy approach as a therapeutic treatment for SCI.

Original language	English (US)
Pages (from-to)	1235-1244
Number of pages	10
Journal	Cytotherapy
Volume	14
Issue number	10
DOIs	https://doi.org/10.3109/14653249.2012.714865
State	Published - Nov 2012

Keywords

NG2
astrocytes
axon regeneration
chondroitin sulfate proteoglycans
glial scar
microglia
neurotrophin-3
spinal cord injury

ASJC Scopus subject areas

Immunology and Allergy
Immunology
Oncology
Genetics(clinical)
Cell Biology
Cancer Research
Transplantation

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Donnelly, E. M., Madigan, N. N., Rooney, G. E., Knight, A., Chen, B., Ball, B., Kinnavane, L., Garcia, Y., Dockery, P., Fraher, J., Strappe, P. M., Windebank, A. J., O'Brien, T., & McMahon, S. S. (2012). Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord. Cytotherapy, 14(10), 1235-1244. https://doi.org/10.3109/14653249.2012.714865

Donnelly, EM, Madigan, NN, Rooney, GE, Knight, A, Chen, B, Ball, B, Kinnavane, L, Garcia, Y, Dockery, P, Fraher, J, Strappe, PM, Windebank, AJ, O'Brien, T & McMahon, SS 2012, 'Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord', Cytotherapy, vol. 14, no. 10, pp. 1235-1244. https://doi.org/10.3109/14653249.2012.714865

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title = "Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord",

abstract = "Background aims. In this study we investigated the effect of neurotrophin-3 (NT-3) and knockdown of NG2, one of the main inhibitory chondroitin sulfate proteoglycans (CSPG), in the glial scar following spinal cord injury (SCI). Methods. Short hairpin (sh) RNA were designed to target NG2 and were cloned into a lentiviral vector (LV). A LV was also constructed containing NT-3. LV expressing NT-3, shRNA to NG2 or combinations of both vectors were injected directly into contused rat spinal cords 1 week post-injury. Six weeks post-injection of LV, spinal cords were examined by histology for changes in scar size and by immunohistochemistry for changes in expression of CSPG, NT-3, astrocytes, neurons and microglia/macrophages. Motor function was assessed using the Basso, Beattie and Bresnahan (BBB) locomotor scale. Results. Animals that received the combination treatment of LV shNG2 and LV NT-3 showed reduced scar size. These animals also showed an increase in levels of neurons and NG2, a decrease in levels of astrocytes and a significant functional recovery as assessed using the BBB locomotor scale at 2 weeks post-treatment. Conclusions. The improvement in locomotor recovery and decrease in scar size shows the potential of this gene therapy approach as a therapeutic treatment for SCI.",

keywords = "NG2, astrocytes, axon regeneration, chondroitin sulfate proteoglycans, glial scar, microglia, neurotrophin-3, spinal cord injury",

author = "Donnelly, {Eleanor M.} and Madigan, {Nicolas N.} and Rooney, {Gemma E.} and Andrew Knight and Bingkun Chen and Bret Ball and Lisa Kinnavane and Yolanda Garcia and Peter Dockery and John Fraher and Strappe, {Padraig M.} and Windebank, {Anthony J.} and Timothy O'Brien and McMahon, {Siobhan S.}",

note = "Funding Information: We wish to thank Professor Dider Trono (EFPL, Switzerland) for the kind gift of the lentiviral vector gene transfer and packaging plasmids. The authors wish to acknowledge the facilities and scientific assistance of the Centre for Microscopy and Imaging at the National University of Ireland Galway ( www.imaging.nuigalway.ie ), a facility that is funded by NUIG and the Irish Government's Programme for Research in third Level Institutions, Cycles 4 and 5, National Development Plan 2007–2013. Funding Information: This work was supported by funding from the Health Research Board of Ireland and Science Foundation Ireland . ",

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doi = "10.3109/14653249.2012.714865",

language = "English (US)",

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T1 - Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord

AU - Donnelly, Eleanor M.

AU - Madigan, Nicolas N.

AU - Rooney, Gemma E.

AU - Knight, Andrew

AU - Chen, Bingkun

AU - Ball, Bret

AU - Kinnavane, Lisa

AU - Garcia, Yolanda

AU - Dockery, Peter

AU - Fraher, John

AU - Strappe, Padraig M.

AU - Windebank, Anthony J.

AU - O'Brien, Timothy

AU - McMahon, Siobhan S.

N1 - Funding Information: We wish to thank Professor Dider Trono (EFPL, Switzerland) for the kind gift of the lentiviral vector gene transfer and packaging plasmids. The authors wish to acknowledge the facilities and scientific assistance of the Centre for Microscopy and Imaging at the National University of Ireland Galway ( www.imaging.nuigalway.ie ), a facility that is funded by NUIG and the Irish Government's Programme for Research in third Level Institutions, Cycles 4 and 5, National Development Plan 2007–2013. Funding Information: This work was supported by funding from the Health Research Board of Ireland and Science Foundation Ireland .

PY - 2012/11

Y1 - 2012/11

N2 - Background aims. In this study we investigated the effect of neurotrophin-3 (NT-3) and knockdown of NG2, one of the main inhibitory chondroitin sulfate proteoglycans (CSPG), in the glial scar following spinal cord injury (SCI). Methods. Short hairpin (sh) RNA were designed to target NG2 and were cloned into a lentiviral vector (LV). A LV was also constructed containing NT-3. LV expressing NT-3, shRNA to NG2 or combinations of both vectors were injected directly into contused rat spinal cords 1 week post-injury. Six weeks post-injection of LV, spinal cords were examined by histology for changes in scar size and by immunohistochemistry for changes in expression of CSPG, NT-3, astrocytes, neurons and microglia/macrophages. Motor function was assessed using the Basso, Beattie and Bresnahan (BBB) locomotor scale. Results. Animals that received the combination treatment of LV shNG2 and LV NT-3 showed reduced scar size. These animals also showed an increase in levels of neurons and NG2, a decrease in levels of astrocytes and a significant functional recovery as assessed using the BBB locomotor scale at 2 weeks post-treatment. Conclusions. The improvement in locomotor recovery and decrease in scar size shows the potential of this gene therapy approach as a therapeutic treatment for SCI.

AB - Background aims. In this study we investigated the effect of neurotrophin-3 (NT-3) and knockdown of NG2, one of the main inhibitory chondroitin sulfate proteoglycans (CSPG), in the glial scar following spinal cord injury (SCI). Methods. Short hairpin (sh) RNA were designed to target NG2 and were cloned into a lentiviral vector (LV). A LV was also constructed containing NT-3. LV expressing NT-3, shRNA to NG2 or combinations of both vectors were injected directly into contused rat spinal cords 1 week post-injury. Six weeks post-injection of LV, spinal cords were examined by histology for changes in scar size and by immunohistochemistry for changes in expression of CSPG, NT-3, astrocytes, neurons and microglia/macrophages. Motor function was assessed using the Basso, Beattie and Bresnahan (BBB) locomotor scale. Results. Animals that received the combination treatment of LV shNG2 and LV NT-3 showed reduced scar size. These animals also showed an increase in levels of neurons and NG2, a decrease in levels of astrocytes and a significant functional recovery as assessed using the BBB locomotor scale at 2 weeks post-treatment. Conclusions. The improvement in locomotor recovery and decrease in scar size shows the potential of this gene therapy approach as a therapeutic treatment for SCI.

KW - NG2

KW - astrocytes

KW - axon regeneration

KW - chondroitin sulfate proteoglycans

KW - glial scar

KW - microglia

KW - neurotrophin-3

KW - spinal cord injury

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Lentiviral vector delivery of short hairpin RNA to NG2 and neurotrophin-3 promotes locomotor recovery in injured rat spinal cord

Abstract

Keywords

ASJC Scopus subject areas

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