Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors

Emily A. Saunderson, Peter Stepper, Jennifer J. Gomm, Lily Hoa, Adrienne Morgan, Michael D. Allen, J. Louise Jones, John G. Gribben, Tomasz P Jurkowski, Gabriella Ficz

Research output: Contribution to journalArticlepeer-review

47 Citations (Scopus)
7 Downloads (Pure)

Abstract

Aberrant promoter DNA hypermethylation is a hallmark of cancer; however, whether this is sufficient to drive cellular transformation is not clear. To investigate this question, we use a CRISPR-dCas9 epigenetic editing tool, where an inactive form of Cas9 is fused to DNA methyltransferase effectors. Using this system, here we show simultaneous de novo DNA methylation of genes commonly methylated in cancer, CDKN2A, RASSF1, HIC1 and PTEN in primary breast cells isolated from healthy human breast tissue. We find that promoter methylation is maintained in this system, even in the absence of the fusion construct, and this prevents cells from engaging senescence arrest. Our data show that the key driver of this phenotype is repression of CDKN2A transcript p16 where myoepithelial cells harbour cancer-like gene expression but do not exhibit anchorage-independent growth. This work demonstrates that hit-and-run epigenetic events can prevent senescence entry, which may facilitate tumour initiation.

Original languageEnglish
Article number1450
JournalNature Communications
Volume8
Issue number1
DOIs
Publication statusPublished - 13 Nov 2017

Keywords

  • Breast/cytology
  • CRISPR-Cas Systems/genetics
  • Cell Transformation, Neoplastic/genetics
  • Cells, Cultured
  • Cellular Senescence/genetics
  • Cyclin-Dependent Kinase Inhibitor p16/metabolism
  • Cyclin-Dependent Kinase Inhibitor p18/genetics
  • DNA (Cytosine-5-)-Methyltransferases/genetics
  • DNA Methylation/genetics
  • Epigenomics
  • Female
  • Gene Editing/methods
  • Humans
  • Kruppel-Like Transcription Factors/genetics
  • PTEN Phosphohydrolase/genetics
  • Promoter Regions, Genetic/genetics
  • Tumor Suppressor Proteins/genetics

Cite this