A native function for RAN translation and CGG repeats in regulating fragile X protein synthesis

Nat Neurosci. 2020 Mar;23(3):386-397. doi: 10.1038/s41593-020-0590-1. Epub 2020 Feb 17.

Abstract

Repeat-associated non-AUG-initiated translation of expanded CGG repeats (CGG RAN) from the FMR1 5'-leader produces toxic proteins that contribute to neurodegeneration in fragile X-associated tremor/ataxia syndrome. Here we describe how unexpanded CGG repeats and their translation play conserved roles in regulating fragile X protein (FMRP) synthesis. In neurons, CGG RAN acts as an inhibitory upstream open reading frame to suppress basal FMRP production. Activation of mGluR5 receptors enhances FMRP synthesis. This enhancement requires both the CGG repeat and CGG RAN initiation sites. Using non-cleaving antisense oligonucleotides (ASOs), we selectively blocked CGG RAN. This ASO blockade enhanced endogenous FMRP expression in human neurons. In human and rodent neurons, CGG RAN-blocking ASOs suppressed repeat toxicity and prolonged survival. These findings delineate a native function for CGG repeats and RAN translation in regulating basal and activity-dependent FMRP synthesis, and they demonstrate the therapeutic potential of modulating CGG RAN translation in fragile X-associated disorders.

Publication types

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

MeSH terms

  • Animals
  • Cell Line
  • Cell Survival / genetics
  • DNA Repeat Expansion / genetics*
  • Female
  • Fragile X Mental Retardation Protein / biosynthesis
  • Fragile X Mental Retardation Protein / genetics*
  • Fragile X Syndrome / genetics*
  • Induced Pluripotent Stem Cells
  • Male
  • Mice
  • Neurons / metabolism
  • Oligonucleotides, Antisense / pharmacology
  • Protein Biosynthesis
  • Rats
  • Rats, Long-Evans
  • Rats, Sprague-Dawley
  • Receptor, Metabotropic Glutamate 5 / biosynthesis
  • Receptor, Metabotropic Glutamate 5 / genetics
  • Trinucleotide Repeats / genetics*

Substances

  • GRM5 protein, human
  • Oligonucleotides, Antisense
  • Receptor, Metabotropic Glutamate 5
  • Fragile X Mental Retardation Protein