Slow Intrathecal Injection of rAAVrh10 Enhances its Transduction of Spinal Cord and Therapeutic Efficacy in a Mutant SOD1 Model of ALS

Neuroscience. 2017 Dec 4:365:192-205. doi: 10.1016/j.neuroscience.2017.10.001. Epub 2017 Oct 9.

Abstract

Mutant SOD1 causes amyotrophic lateral sclerosis (ALS) by a dominant gain of toxicity. Previous studies have demonstrated therapeutic potential of mutant SOD1-RNAi delivered by intrathecal (IT) injection of recombinant adeno-associated virus (rAAV). However, optimization of delivery is needed to overcome the high degree of variation in the transduction efficiency and therapeutic efficacy. Here, on the basis of our previously defined, efficient IT injection method, we investigated the influence of injection speed on transduction efficiency in the central nervous system (CNS). We demonstrate that slow IT injection results in higher transduction of spinal cord and dorsal root ganglia (DRG) while fast IT injection leads to higher transduction of brain and peripheral organs. To test how these effects influence the outcome of RNAi therapy, we used slow and fast IT injection to deliver rAAVrh10-GFP-amiR-SOD1, a rAAV vector that expresses GFP and an artificial miRNA targeting SOD1, in SOD1-G93A mice. Both slow and fast IT injection produced therapeutic efficacy but the slow injection trended slightly toward a better outcome than the fast injection. These results demonstrate that IT injection speed influences the predominance of gene delivery at different CNS sites and should be taken into consideration in future therapeutic trials involving IT injection.

Keywords: AAV; SOD1-G93A; amyotrophic lateral sclerosis; gene therapy; intrathecal.

MeSH terms

  • Adenomatous Polyposis Coli Protein / metabolism
  • Amyotrophic Lateral Sclerosis* / genetics
  • Amyotrophic Lateral Sclerosis* / pathology
  • Amyotrophic Lateral Sclerosis* / therapy
  • Animals
  • Body Weight / genetics
  • Calcium-Binding Proteins / metabolism
  • Dependovirus / genetics
  • Disease Models, Animal
  • Ganglia, Spinal / metabolism
  • Ganglia, Spinal / pathology
  • Genetic Therapy / methods*
  • Glial Fibrillary Acidic Protein / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Injections, Spinal
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microfilament Proteins / metabolism
  • Mutation / genetics*
  • Spinal Cord / metabolism*
  • Spinal Cord / pathology
  • Superoxide Dismutase / genetics*
  • Transduction, Genetic

Substances

  • Adenomatous Polyposis Coli Protein
  • Aif1 protein, mouse
  • Calcium-Binding Proteins
  • Glial Fibrillary Acidic Protein
  • Microfilament Proteins
  • adenomatous polyposis coli protein, mouse
  • Green Fluorescent Proteins
  • SOD1 G93A protein
  • Superoxide Dismutase