Abstract
The gene therapy has been successful in treatment of spinal cord injury (SCI) in several animal models, although it still remains unavailable for clinical practice. Surprisingly, regardless the fact that multiple reports showed motor recovery with gene therapy, little is known about molecular and cellular changes in the post-traumatic spinal cord following viral vector- or cell-mediated gene therapy. In this study we evaluated the therapeutic efficacy and changes in spinal cord after treatment with the genes encoding vascular endothelial growth factor (VEGF), glial cell-derived neurotrophic factor (GDNF), angiogenin (ANG), and neuronal cell adhesion molecule (NCAM) applied using both approaches. Therapeutic genes were used for viral vector- and cell-mediated gene therapy in two combinations: (1) VEGF+GDNF+NCAM and (2) VEGF+ANG+NCAM. For direct gene therapy adenoviral vectors based on serotype 5 (Ad5) were injected intrathecally and for cell-mediated gene delivery human umbilical cord blood mononuclear cells (UCB-MC) were simultaneously transduced with three Ad5 vectors and injected intrathecally 4 h after the SCI. The efficacy of both treatments was confirmed by improvement in behavioral (BBB) test. Molecular and cellular changes following post-traumatic recovery were evaluated with immunofluorescent staining using antibodies against the functional markers of motorneurons (Hsp27, synaptophysin, PSD95), astrocytes (GFAP, vimentin), oligodendrocytes (Olig2, NG2, Cx47) and microglial cells (Iba1). Our results suggest that both approaches with intrathecal delivery of therapeutic genes may support functional recovery of post-traumatic spinal cord via lowering the stress (down regulation of Hsp25) and enhancing the synaptic plasticity (up regulation of PSD95 and synaptophysin), supporting oligodendrocyte proliferation (up regulation of NG2) and myelination (up regulation of Olig2 and Cx47), modulating astrogliosis by reducing number of astrocytes (down regulation of GFAP and vimetin) and microglial cells (down regulation of Iba1).
Highlights
To date the interest to the gene therapy is growing and motivated by results from animal research that show a potential for chronic spinal cord injury (SCI)
Concentrations of vascular endothelial growth factor (VEGF) measured in conditioned medium of umbilical cord blood mononuclear cells (UCB-MC)+adenoviral vectors (Ad)-VEGF-glial cell-derived neurotrophic factor (GDNF)-neuronal cell adhesion molecule (NCAM) and UCBMCs+Ad-green fluorescent protein (GFP) groups were 724.3 and 181.8 pg/ml, respectively
The GFP fluorescence in UCB-MCs was evident for a month after cells transduction with adenoviral vectors based on serotype 5 (Ad5)-GFP
Summary
Several therapeutic genes are considered to be effective for single gene therapy, i.e., encoding neurotrophic BDNF (Nakajima et al, 2007), NGF (Romero et al, 2001), GDNF (Chou et al, 2014), growth VEGF, FGF (De Laporte et al, 2011), Angiogenin (Povysheva et al, 2017), anti-apoptotic BCL-2, BCL-XL (Yukawa et al, 2002), and anti-inflammatory IL-10, IL-1RA (Zhou et al, 2009) factors, and cell adhesion NCAM and L1 molecules (Chaudhry et al, 2006; Thuret et al, 2006; Walthers and Seidlits, 2015). Along with other findings, we observed improvements and prolongation of lifespan in amyotrophic lateral sclerosis (ALS) mice after transplantation of the umbilical cord blood mononuclear cells (UCB-MCs) simultaneously transduced with three adenoviral vectors (Ad) encoding vascular endothelial growth factor (VEGF), glial derived neurotrophic factor (GDNF) and neural cell adhesion molecule (NCAM). This study was focused on the following factors: Vascular Endothelial Growth Factor (VEGF)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
