Title: Single-cell microarray enables high-throughput evaluation of DNA double-strand breaks and DNA repair inhibitors.
Authors: Weingeist, David M; Ge, Jing; Wood, David K; Mutamba, James T; Huang, Qiuying; Rowland, Elizabeth A; Yaffe, Michael B; Floyd, Scott; Engelward, Bevin P
Published In Cell Cycle, (2013 Mar 15)
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.
PubMed ID: 23422001
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