Induction and reversal of myotonic dystrophy type 1 pre-mRNA splicing defects by small molecules

Nat Commun. 2013:4:2044. doi: 10.1038/ncomms3044.

Abstract

The ability to control pre-mRNA splicing with small molecules could facilitate the development of therapeutics or cell-based circuits that control gene function. Myotonic dystrophy type 1 is caused by the dysregulation of alternative pre-mRNA splicing due to sequestration of muscleblind-like 1 protein (MBNL1) by expanded, non-coding r(CUG) repeats (r(CUG)(exp)). Here we report two small molecules that induce or ameliorate alternative splicing dysregulation. A thiophene-containing small molecule (1) inhibits the interaction of MBNL1 with its natural pre-mRNA substrates. Compound (2), a substituted naphthyridine, binds r(CUG)(exp) and displaces MBNL1. Structural models show that 1 binds MBNL1 in the Zn-finger domain and that 2 interacts with UU loops in r(CUG)(exp). This study provides a structural framework for small molecules that target MBNL1 by mimicking r(CUG)(exp) and shows that targeting MBNL1 causes dysregulation of alternative splicing, suggesting that MBNL1 is thus not a suitable therapeutic target for the treatment of myotonic dystrophy type 1.

Publication types

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

MeSH terms

  • Animals
  • Base Sequence
  • Exons / genetics
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Gene Knockdown Techniques
  • HEK293 Cells
  • HeLa Cells
  • High-Throughput Screening Assays
  • Humans
  • Mice
  • Molecular Docking Simulation
  • Molecular Sequence Data
  • Myotonic Dystrophy / drug therapy*
  • Myotonic Dystrophy / genetics*
  • Plasmids / metabolism
  • Protein Binding / genetics
  • Protein Biosynthesis / drug effects
  • RNA / metabolism
  • RNA Precursors / genetics*
  • RNA Precursors / metabolism
  • RNA Splicing / drug effects
  • RNA Splicing / genetics*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism
  • Receptor, Insulin / genetics
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology
  • Small Molecule Libraries / therapeutic use*

Substances

  • MBNL1 protein, human
  • RNA Precursors
  • RNA-Binding Proteins
  • Small Molecule Libraries
  • RNA
  • Receptor, Insulin