Twist2 amplification in rhabdomyosarcoma represses myogenesis and promotes oncogenesis by redirecting MyoD DNA binding

Genes Dev. 2019 Jun 1;33(11-12):626-640. doi: 10.1101/gad.324467.119. Epub 2019 Apr 11.


Rhabdomyosarcoma (RMS) is an aggressive pediatric cancer composed of myoblast-like cells. Recently, we discovered a unique muscle progenitor marked by the expression of the Twist2 transcription factor. Genomic analyses of 258 RMS patient tumors uncovered prevalent copy number amplification events and increased expression of TWIST2 in fusion-negative RMS. Knockdown of TWIST2 in RMS cells results in up-regulation of MYOGENIN and a decrease in proliferation, implicating TWIST2 as an oncogene in RMS. Through an inducible Twist2 expression system, we identified Twist2 as a reversible inhibitor of myogenic differentiation with the remarkable ability to promote myotube dedifferentiation in vitro. Integrated analysis of genome-wide ChIP-seq and RNA-seq data revealed the first dynamic chromatin and transcriptional landscape of Twist2 binding during myogenic differentiation. During differentiation, Twist2 competes with MyoD at shared DNA motifs to direct global gene transcription and repression of the myogenic program. Additionally, Twist2 shapes the epigenetic landscape to drive chromatin opening at oncogenic loci and chromatin closing at myogenic loci. These epigenetic changes redirect MyoD binding from myogenic genes toward oncogenic, metabolic, and growth genes. Our study reveals the dynamic interplay between two opposing transcriptional regulators that control the fate of RMS and provides insight into the molecular etiology of this aggressive form of cancer.

Keywords: bHLH; chromosome amplification; dedifferentiation; histone modification; rhabdomyosarcoma; skeletal muscle.

Publication types

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

MeSH terms

  • Carcinogenesis*
  • Cells, Cultured
  • Chromatin Assembly and Disassembly
  • DNA / metabolism
  • Epithelial-Mesenchymal Transition
  • Gene Amplification
  • Gene Expression Regulation, Neoplastic
  • HEK293 Cells
  • Helix-Loop-Helix Motifs
  • Humans
  • Muscle Development*
  • MyoD Protein / chemistry
  • MyoD Protein / metabolism*
  • Myoblasts / metabolism
  • Nuclear Proteins / genetics
  • Repressor Proteins / chemistry
  • Repressor Proteins / genetics*
  • Repressor Proteins / metabolism*
  • Rhabdomyosarcoma / genetics*
  • Rhabdomyosarcoma / metabolism*
  • Twist-Related Protein 1 / chemistry
  • Twist-Related Protein 1 / genetics*
  • Twist-Related Protein 1 / metabolism*


  • MyoD Protein
  • MyoD1 myogenic differentiation protein
  • Nuclear Proteins
  • Repressor Proteins
  • TWIST1 protein, human
  • TWIST2 protein, human
  • Twist-Related Protein 1
  • DNA