A synthetic lethal siRNA screen identifying genes mediating sensitivity to a PARP inhibitor

EMBO J. 2008 May 7;27(9):1368-77. doi: 10.1038/emboj.2008.61. Epub 2008 Apr 3.

Abstract

Inhibitors of poly (ADP-ribose)-polymerase-1 (PARP) are highly lethal to cells with deficiencies in BRCA1, BRCA2 or other components of the homologous recombination pathway. This has led to PARP inhibitors entering clinical trials as a potential therapy for cancer in carriers of BRCA1 and BRCA2 mutations. To discover new determinants of sensitivity to these drugs, we performed a PARP-inhibitor synthetic lethal short interfering RNA (siRNA) screen. We identified a number of kinases whose silencing strongly sensitised to PARP inhibitor, including cyclin-dependent kinase 5 (CDK5), MAPK12, PLK3, PNKP, STK22c and STK36. How CDK5 silencing mediates sensitivity was investigated. Previously, CDK5 has been suggested to be active only in a neuronal context, but here we show that CDK5 is required in non-neuronal cells for the DNA-damage response and, in particular, intra-S and G(2)/M cell-cycle checkpoints. These results highlight the potential of synthetic lethal siRNA screens with chemical inhibitors to define new determinants of sensitivity and potential therapeutic targets.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cell Cycle / drug effects
  • Cell Line, Tumor
  • Cyclin-Dependent Kinase 5 / genetics
  • Cyclin-Dependent Kinase 5 / metabolism
  • DNA Damage
  • DNA Repair
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • HeLa Cells
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Mitogen-Activated Protein Kinase 12 / genetics
  • Mitogen-Activated Protein Kinase 12 / metabolism
  • Poly(ADP-ribose) Polymerase Inhibitors*
  • Poly(ADP-ribose) Polymerases / metabolism
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / metabolism
  • RNA, Small Interfering / genetics*
  • Transfection

Substances

  • Poly(ADP-ribose) Polymerase Inhibitors
  • RNA, Small Interfering
  • Hydrogen Peroxide
  • Poly(ADP-ribose) Polymerases
  • Mitogen-Activated Protein Kinase 12
  • PLK3 protein, human
  • Cyclin-Dependent Kinase 5
  • Protein-Serine-Threonine Kinases
  • STK36 protein, human