Cis-regulatory elements driving motor neuron-selective viral payload expression within the mammalian spinal cord

Proc Natl Acad Sci U S A. 2024 Dec 3;121(49):e2418024121. doi: 10.1073/pnas.2418024121. Epub 2024 Nov 27.

Abstract

Spinal motor neuron (MN) dysfunction is the cause of a number of clinically significant movement disorders. Despite the recent approval of gene therapeutics targeting these MN-related disorders, there are no viral delivery mechanisms that achieve MN-restricted transgene expression. In this study, chromatin accessibility profiling of genetically defined mouse MNs was used to identify candidate cis-regulatory elements (CREs) capable of driving MN-selective gene expression. Subsequent testing of these candidates identified two CREs that confer MN-selective gene expression in the spinal cord as well as reduced off-target expression in dorsal root ganglia. Within one of these candidate elements, we identified a compact core transcription factor (TF)-binding region that drives MN-selective gene expression. Finally, we demonstrated that selective spinal cord expression driven by this mouse CRE is preserved in non-human primates. These findings suggest that cell-type-selective viral reagents in which cell-type-selective CREs drive restricted gene expression will be valuable research tools in mice and other mammalian species, with potentially significant therapeutic value in humans.

Keywords: AAV; cis-regulatory element; dorsal root ganglion; gene therapy; motor neuron.

MeSH terms

  • Animals
  • Ganglia, Spinal / metabolism
  • Ganglia, Spinal / virology
  • Genetic Vectors / genetics
  • Humans
  • Mice
  • Motor Neurons* / metabolism
  • Motor Neurons* / virology
  • Regulatory Sequences, Nucleic Acid / genetics
  • Spinal Cord* / metabolism
  • Spinal Cord* / virology
  • Transgenes