Gene delivery to the spinal cord: comparison between lentiviral, adenoviral, and retroviral vector delivery systems.
Viral gene delivery for spinal cord injury (SCI) is a promising approach for enhancing axonal regeneration and neuroprotection. An understanding of spatio-temporal transgene expression in the spinal cord is essential for future studies of SCI therapies. Commonly, intracellular marker proteins (e.g., EGFP) were used as indicators of transgene levels after viral delivery, which may not accurately reflect levels of secreted transgene. This study examined transgene expression using ELISA after viral delivery of D15A, a neurotrophin with BDNF and NT-3 activities, at 1, 2, and 4weeks after in vivo and ex vivo delivery using lentiviral, adenoviral, and retroviral vectors. Further, the inflammatory responses and viral infection patterns after in vivo delivery were examined. Lentiviral vectors had the most stable pattern of gene expression, with D15A levels of 536 +/- 38 and 363 +/- 47 pg/mg protein seen at 4 weeks after the in vivo and ex vivo delivery, respectively. Our results show that protein levels downregulate disproportionately to levels of EGFP after adenoviral vectors both in vivo and ex vivo. D15A dropped from initial levels of 422 +/- 87 to 153 +/- 18 pg/mg protein at 4 weeks after in vivo administration. Similarly, ex vivo retrovirus-mediated transgene expression exhibited rapid downregulation by 2 weeks post-grafting. Compared to adenoviral infection, macrophage activation was attenuated after lentiviral infection. These results suggest that lentiviral vectors are most suitable in situations where stable long-term transgene expression is needed. Retroviral ex vivo delivery is optional when transient expression within targeted spinal tissue is desired, with adenoviral vectors in between.
Abdellatif
Ahmed A
AA
Department of Anatomical Sciences and Neurobiology, University of Louisville, School of Medicine, Louisville, Kentucky 40202, USA.
Pelt
Jennifer L
JL
Benton
Richard L
RL
Howard
Russell M
RM
Tsoulfas
Pantelis
P
Ping
Peipei
P
Xu
Xiao-Ming
XM
Whittemore
Scott R
SR
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D015870:Gene Expression / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics* / Q000627:therapeutic use; D006085:Graft Survival / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D009187:Myelitis / Q000378:metabolism / Q000517:prevention & control / Q000821:virology; D041681:NIH 3T3 Cells; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism; D009416:Nerve Regeneration / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012190:Retroviridae / Q000235:genetics; D013116:Spinal Cord / Q000378:metabolism* / Q000601:surgery / Q000821:virology; D013119:Spinal Cord Injuries / Q000628:therapy*; D016378:Tissue Transplantation / Q000379:methods; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics
D009414:Nerve Growth Factors