Intrathecal Vector Delivery in Juvenile Rats via Lumbar Cistern Injection

Area of Science:

• Neuroscience
• Gene Therapy
• Molecular Biology

Background:

• Gene therapy offers potential treatments for diseases by introducing, inactivating, or modulating genes.
• Targeting the central nervous system (CNS) for neurodegenerative disorders often requires efficient gene delivery to the brain and spinal cord.
• Cerebrospinal fluid (CSF) delivery via lumbar intrathecal injection is a minimally invasive method to achieve widespread CNS transduction.

Purpose of the Study:

• To describe a refined method for performing lumbar puncture in juvenile rats for cerebrospinal fluid (CSF) delivery of gene therapy vectors.
• To evaluate the transduction efficiency and profile following lumbar intrathecal injection in a juvenile rat model.

Main Methods:

• A lumbar puncture technique was employed to inject adeno-associated virus serotype 9 (AAV9) vectors into the lumbar cistern of juvenile rats.
• Injections utilized 25-35 µL of vector, with a green fluorescent protein (GFP) reporter gene to visualize transduction.
• The procedure was adapted for the specific anatomical and physiological characteristics of juvenile rats.

Main Results:

• Successful delivery of gene therapy vectors into the CSF via lumbar puncture was achieved in juvenile rats.
• The green fluorescent protein (GFP) reporter demonstrated successful transduction within the central nervous system (CNS).
• The study identified benefits and challenges associated with this specific gene delivery approach in the target species.

Conclusions:

• Lumbar intrathecal injection provides a viable and minimally invasive route for gene therapy vector delivery to the CNS in juvenile rats.
• This method holds promise for preclinical studies of neurodegenerative diseases and other CNS disorders.
• Further optimization may enhance the safety and efficacy of gene therapy delivery to the juvenile CNS.