Prospects of siRNA cocktails as tools for modifying multiple gene targets in the injured spinal cord.

Unlabelled: Gene silencing through RNA interference (RNAi) has been touted as a boon for identifying potential therapies for difficult-to-treat pathologies. In this regard, siRNA-mediated gene silencing for tackling the multifaceted pathophysiology of spinal cord injury seemed promising. The genes caspase 3 and sarm1 were targeted in the present study, using siRNAs in a rodent model of spinal cord injury, as the feasibility of concomitant silencing of more than one gene had not been previously attempted. The results indicated meager benefits in terms of functional recovery and tissue preservation. Interestingly, differential transfection efficiencies due to the heterogeneous nature of cells in the spinal cord along with variability in efficacy based on time of intervention affected the reproducibility of this approach. Complex gene interactions and inadequacies in molecular evaluation strategies further complicated the interpretation of the outcome. If these glitches are resolved through further research, gene therapy in general and RNAi, in particular, may become a mainstay approach for treating contusion spinal cord injury.

Impact Statement: Gene therapy has reached the level of clinical trials. However, safety and efficacy are yet to be confirmed. The present study tested the prospects of gene silencing using siRNAs in a rat model of spinal cord injury. Some noteworthy observations include the effective and long-lasting silencing effects of siRNAs, inhibition of one gene's expression resulting in silencing of multiple genes in associated pathways, possibility of targeting more than one gene through siRNA cocktails, and differential gene silencing effects based on temporal changes in their expression patterns. It is argued that differential uptake of siRNAs by cells as observed and limitations in the analysis methods available can skew interpretations. Thus, this study may serve as a cautionary tale indicating that gene silencing using siRNAs for spinal cord injury can be a potential therapy, but practical issues are to be addressed in order to ensure consistency and safety.