Ex Vivo Rat Transected Spinal Cord Slices as a Model to Assess Lentiviral Vector Delivery of Neurotrophin-3 and Short Hairpin RNA against NG2

Biology, 2020 · DOI: 10.3390/biology9030054 · Published: March 15, 2020

Simple Explanation

This study uses an ex vivo model of spinal cord injury to test a gene therapy approach. The approach involves delivering Neurotrophin-3 (NT-3) and short hairpin RNA against NG2 (NG2 sh) using lentiviral vectors. The researchers found that combining NT-3 and NG2 sh promoted axonal growth in injured spinal cord slices. This suggests that the combination of these two factors can create a more favorable environment for neuronal regeneration after spinal cord injury. The study also highlights the potential of using ex vivo organotypic slice cultures (OSCs) as a platform for studying glial scarring and evaluating potential treatments for spinal cord injury.

Study Duration

Not specified

Participants

Sprague Dawley rats (P4 pups)

Evidence Level

In vitro and ex vivo study

Key Findings

1
Conditioned medium from cells transduced with NT-3 or shNG2 lentiviruses caused a significant increase in neurite length of primary dorsal root ganglia neurons compared to the control group in vitro.
2
In an ex vivo organotypic slice culture (OSC) transduction with Lenti-NT-3 promoted axonal growth.
3
Transducing OSCs with a combination of Lenti-NT-3/NG2 sh lead to a further increase in axonal growth but only in injured slices and only within the region adjacent to the site of injury.

Research Summary

This study investigated the eﬀect of lentiviral vectors expressing Neurotrophin-3 (NT-3) and short-hairpin RNA against NG2 (NG2 sh) to enhance neurite outgrowth in in vitro and ex vivo transection injury models. These ﬁndings suggest that the combination of lentiviral NT-3 and NG2 sh reduced NG2 levels and provided a more favourable microenvironment for neuronal regeneration after SCI. This study also shows that OSCs may be a useful platform for studying glial scarring and potential SCI treatments.

Practical Implications

Therapeutic Target

Combined lentiviral NT-3 and NG2 sh gene therapy shows promise for treating spinal cord injury by promoting axonal regrowth and reducing glial scarring.

Research Tool

Ex vivo OSCs are a useful platform for studying glial scarring and evaluating potential SCI treatments, potentially reducing the need for in vivo animal models.

Drug Delivery

Lentiviral vectors can efficiently deliver therapeutic genes (NT-3) and shRNA (against NG2) to spinal cord tissue, suggesting a potential strategy for targeted gene therapy in SCI.

Study Limitations

1
The ex vivo OSC lacks a blood supply, limiting the assessment of circulating immune cells' effects.
2
The study used slices from newborn (P4) pup spinal cords, which may not reflect SCI responses in adult animals.
3
Functional and behavioral assessments cannot be conducted using this system.

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