DNAzyme Cleavage of CAG Repeat RNA in Polyglutamine Diseases

Neurotherapeutics. 2021 Jul;18(3):1710-1728. doi: 10.1007/s13311-021-01075-w. Epub 2021 Jun 23.

Abstract

CAG repeat expansion is the genetic cause of nine incurable polyglutamine (polyQ) diseases with neurodegenerative features. Silencing repeat RNA holds great therapeutic value. Here, we developed a repeat-based RNA-cleaving DNAzyme that catalyzes the destruction of expanded CAG repeat RNA of six polyQ diseases with high potency. DNAzyme preferentially cleaved the expanded allele in spinocerebellar ataxia type 1 (SCA1) cells. While cleavage was non-allele-specific for spinocerebellar ataxia type 3 (SCA3) cells, treatment of DNAzyme leads to improved cell viability without affecting mitochondrial metabolism or p62-dependent aggresome formation. DNAzyme appears to be stable in mouse brain for at least 1 month, and an intermediate dosage of DNAzyme in a SCA3 mouse model leads to a significant reduction of high molecular weight ATXN3 proteins. Our data suggest that DNAzyme is an effective RNA silencing molecule for potential treatment of multiple polyQ diseases.

Keywords: CAG repeats; DNAzyme; Huntington’s disease; Microsatellite expansion; Polyglutamine; Spinocerebellar ataxia.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ataxin-3 / genetics
  • Cell Line, Tumor
  • DNA, Catalytic / administration & dosage*
  • DNA, Catalytic / genetics*
  • Gene Silencing / physiology
  • HEK293 Cells
  • Humans
  • Machado-Joseph Disease / genetics*
  • Machado-Joseph Disease / therapy
  • Mice
  • Peptides / genetics*
  • Peptides / metabolism
  • RNA / genetics*
  • Spinocerebellar Ataxias / genetics
  • Spinocerebellar Ataxias / therapy
  • Stereotaxic Techniques
  • Trinucleotide Repeat Expansion / genetics*

Substances

  • DNA, Catalytic
  • Peptides
  • polyglutamine
  • RNA
  • Ataxin-3
  • Atxn3 protein, mouse