TY - JOUR
T1 - Engineering polysialic acid on Schwann cells using polysialyltransferase gene transfer or purified enzyme exposure for spinal cord injury transplantation
AU - Pearse, Damien D.
AU - Rao, Sudheendra N.R.
AU - Morales, Alejo A.
AU - Wakarchuk, Warren
AU - Rutishauser, Urs
AU - El-Maarouf, Abderrahman
AU - Ghosh, Mousumi
N1 - Funding Information:
This research was supported by The Rehabilitation Research & Development Service of the Veteran's Health Administration to D.D.P. (I01RX001050) and M.G. (I01RX002099), The John M. and Jocelyn H.K. Watkins Distinguished Chair in Cell Therapies (D.D.P.), The Miami Project to Cure Paralysis and The Buoniconti Fund (D.D.P. and M.G.). W.W. acknowledges funding from the Canadian Glycomics Network for development of PSTNm. The authors would like to thank the Animal Core Facility at the Miami Project to Cure Paralysis for assisting with animal care, the Miami Project Viral Vector Core for the production of Lentivirus and the Miami Project Imaging Core for assistance with imaging. Leopold Puzis, Weijun An, Alexander Shaheen and Johana De Luca are thanked for their assistance with molecular biology and plasmid generation. Yong Xu is acknowledged for help with surgery and cell transplantation.
Funding Information:
This research was supported by The Rehabilitation Research & Development Service of the Veteran’s Health Administration to D.D.P. ( I01RX001050 ) and M.G. ( I01RX002099 ), The John M. and Jocelyn H.K. Watkins Distinguished Chair in Cell Therapies (D.D.P.), The Miami Project to Cure Paralysis and The Buoniconti Fund (D.D.P. and M.G.). W.W. acknowledges funding from the Canadian Glycomics Network for development of PSTNm. The authors would like to thank the Animal Core Facility at the Miami Project to Cure Paralysis for assisting with animal care, the Miami Project Viral Vector Core for the production of Lentivirus and the Miami Project Imaging Core for assistance with imaging. Leopold Puzis, Weijun An, Alexander Shaheen and Johana De Luca are thanked for their assistance with molecular biology and plasmid generation. Yong Xu is acknowledged for help with surgery and cell transplantation.
Publisher Copyright:
© 2021
PY - 2021/3/23
Y1 - 2021/3/23
N2 - Polysialic acid (PolySia) is a critical post-translational modification on the neural cell adhesion molecule (NCAM, a.k.a., CD56), important for cell migration and axon growth during nervous system development, plasticity and repair. PolySia induction on Schwann cells (SCs) enhances their migration, axon growth support and ability to improve functional recovery after spinal cord injury (SCI) transplantation. In the current investigation two methods of PolySia induction on SCs, lentiviral vector transduction of the mouse polysialytransferase gene ST8SIA4 (LV-PST) or enzymatic engineering with a recombinant bacterial PST (PSTNm), were examined comparatively for their effects on PolySia induction, SC migration, the innate immune response and axon growth after acute SCI. PSTNm produced significant PolySia induction and a greater diversity of surface molecule polysialylation on SCs as evidenced by immunoblot. In the scratch wound assay, PSTNm was superior to LV-PST in the promotion of SC migration and gap closure. At 24 h after SCI transplantation, PolySia induction on SCs was most pronounced with LV-PST. Co-delivery of PSTNm with SCs, but not transient cell exposure, led to broader induction of PolySia within the injured spinal cord due to polysialylation upon both host cells and transplanted SCs. The innate immune response after SCI, measured by CD68 immunoreactivity, was similar among PolySia induction methods. LV-PST or PSTNm co-delivery with SCs provided a similar enhancement of SC migration and axon growth support above that of unmodified SCs. These studies demonstrate that LV-PST and PSTNm provide comparable acute effects on SC polysialation, the immune response and neurorepair after SCI.
AB - Polysialic acid (PolySia) is a critical post-translational modification on the neural cell adhesion molecule (NCAM, a.k.a., CD56), important for cell migration and axon growth during nervous system development, plasticity and repair. PolySia induction on Schwann cells (SCs) enhances their migration, axon growth support and ability to improve functional recovery after spinal cord injury (SCI) transplantation. In the current investigation two methods of PolySia induction on SCs, lentiviral vector transduction of the mouse polysialytransferase gene ST8SIA4 (LV-PST) or enzymatic engineering with a recombinant bacterial PST (PSTNm), were examined comparatively for their effects on PolySia induction, SC migration, the innate immune response and axon growth after acute SCI. PSTNm produced significant PolySia induction and a greater diversity of surface molecule polysialylation on SCs as evidenced by immunoblot. In the scratch wound assay, PSTNm was superior to LV-PST in the promotion of SC migration and gap closure. At 24 h after SCI transplantation, PolySia induction on SCs was most pronounced with LV-PST. Co-delivery of PSTNm with SCs, but not transient cell exposure, led to broader induction of PolySia within the injured spinal cord due to polysialylation upon both host cells and transplanted SCs. The innate immune response after SCI, measured by CD68 immunoreactivity, was similar among PolySia induction methods. LV-PST or PSTNm co-delivery with SCs provided a similar enhancement of SC migration and axon growth support above that of unmodified SCs. These studies demonstrate that LV-PST and PSTNm provide comparable acute effects on SC polysialation, the immune response and neurorepair after SCI.
KW - Axon growth
KW - Cell migration
KW - Polysialic acid
KW - Polysialytransferase
KW - Schwann cell transplantation
KW - Spinal cord injury
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UR - http://www.scopus.com/inward/citedby.url?scp=85100910278&partnerID=8YFLogxK
U2 - 10.1016/j.neulet.2021.135690
DO - 10.1016/j.neulet.2021.135690
M3 - Article
C2 - 33540059
AN - SCOPUS:85100910278
VL - 748
JO - Neuroscience Letters
JF - Neuroscience Letters
SN - 0304-3940
M1 - 135690
ER -