Single-cell microarray enables high-throughput evaluation of DNA double-strand breaks and DNA repair inhibitors

Cell Cycle. 2013 Mar 15;12(6):907-15. doi: 10.4161/cc.23880. Epub 2013 Feb 19.

Abstract

A key modality of non-surgical cancer management is DNA damaging therapy that causes DNA double-strand breaks that are preferentially toxic to rapidly dividing cancer cells. Double-strand break repair capacity is recognized as an important mechanism in drug resistance and is therefore a potential target for adjuvant chemotherapy. Additionally, spontaneous and environmentally induced DSBs are known to promote cancer, making DSB evaluation important as a tool in epidemiology, clinical evaluation and in the development of novel pharmaceuticals. Currently available assays to detect double-strand breaks are limited in throughput and specificity and offer minimal information concerning the kinetics of repair. Here, we present the CometChip, a 96-well platform that enables assessment of double-strand break levels and repair capacity of multiple cell types and conditions in parallel and integrates with standard high-throughput screening and analysis technologies. We demonstrate the ability to detect multiple genetic deficiencies in double-strand break repair and evaluate a set of clinically relevant chemical inhibitors of one of the major double-strand break repair pathways, non-homologous end-joining. While other high-throughput repair assays measure residual damage or indirect markers of damage, the CometChip detects physical double-strand breaks, providing direct measurement of damage induction and repair capacity, which may be useful in developing and implementing treatment strategies with reduced side effects.

Keywords: DNA double-strand breaks; DNA repair; DNA-PK inhibitors; high throughput; microarray; neutral comet assay; neutral single-cell electrophoresis assay; non-homologous end-joining.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • CHO Cells
  • Cell Line
  • Chromones / pharmacology
  • Cricetinae
  • DNA Breaks, Double-Stranded / drug effects*
  • DNA Damage
  • DNA Repair / drug effects*
  • DNA-Activated Protein Kinase / antagonists & inhibitors
  • Drug Resistance, Neoplasm
  • Enzyme Inhibitors / pharmacology
  • High-Throughput Screening Assays / methods*
  • Humans
  • Morpholines / pharmacology
  • Neoplasms / drug therapy*
  • Neoplasms / genetics

Substances

  • 2-(morpholin-4-yl)benzo(h)chromen-4-one
  • 8-dibenzothiophen-4-yl-2-morpholin-4-yl-chromen-4-one
  • Chromones
  • Enzyme Inhibitors
  • Morpholines
  • 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
  • DNA-Activated Protein Kinase