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Differential disruption of EWS-FLI1 binding by DNA-binding agents

Changmin Chen1, Diane R Wonsey, Madeleine E Lemieux

  • 1Department of Pediatric Oncology, Dana-Farber Cancer Institute and Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.

Plos One
|July 30, 2013
PubMed

Insights

Actinomycin D selectively disrupts EWS-FLI1 binding to DNA in Ewing sarcoma cells. While not therapeutically viable due to narrow concentration windows, this highlights potential for targeted transcription factor disruption.

Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • Ewing sarcoma arises from the EWS-FLI1 fusion oncoprotein, which drives abnormal gene expression.
  • Targeting the EWS-FLI1 oncoprotein is crucial for Ewing sarcoma treatment.

Purpose of the Study:

  • To screen for compounds that disrupt EWS-FLI1 binding to DNA.
  • To investigate the preferential binding disruption of EWS-FLI1 by actinomycin D.

Main Methods:

  • Utilized a homogenous proximity assay to screen for DNA-binding disrupting compounds.
  • Evaluated compound effects on EWS-FLI1 and p53 DNA binding in vitro.
  • Assessed actinomycin D's impact on EWS-FLI1 chromatin binding and gene expression in cell-based assays.

Main Results:

  • Actinomycin D preferentially disrupted EWS-FLI1 DNA binding over p53 binding in vitro.
  • Low concentrations of actinomycin D selectively blocked EWS-FLI1 chromatin binding and downstream gene expression.
  • Higher concentrations of actinomycin D caused global transcriptional repression.

Conclusions:

  • Actinomycin D exhibits preferential disruption of EWS-FLI1 DNA binding at specific concentrations.
  • The narrow therapeutic window limits actinomycin D's direct application but suggests potential for base-preference-driven targeted therapies.
  • Exploiting DNA base preferences could lead to compounds that selectively inhibit specific transcription factor subclasses.

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