title: Assessment of the in vitro γH2AX assay by High Content Screening as a novel genotoxicity test.
abstract: The γH2AX assay is widely used as a marker of DNA damage in multiple scientific fields such as cancer biomarker, clinical studies and radiation biology. In particular, the in vitro γH2AX assay has been suggested as a novel in vitro genotoxicity test with potential as a pre-screening tool. However, to date, limited assessments have been carried out to evaluate the sensitivity, specificity and accuracy of the in vitro γH2AX assay. In this study, the microscopy-based system combining automated cellular image acquisition with software quantification for High Content Screening (HCS) has been used for the first time to evaluate the in vitro γH2AX assay. A panel of well-characterised genotoxic and non-genotoxic compounds was selected to assess the performance of the in vitro γH2AX assay in the human bronchial epithelial cell line BEAS-2B. The results obtained during this preliminary assessment indicate that the in vitro γH2AX assay has a high accuracy (86%) as a result of high sensitivity and specificity (86-92% and 80-88% respectively). Our data highlight the potential for γH2AX detection in HCS as a complement to the current regulatory genotoxicity battery of in vitro assays. We therefore recommend more comprehensive assessments to confirm the performance of the in vitro γH2AX assay by HCS with a more extensive set of compounds.
authors.affiliation: British American Tobacco, Group Research and Development, Regents Park Road, Southampton, Hampshire SO15 8TL, United Kingdom; Department of Toxicology and Pharmacology, Universidad Complutense de Madrid, Madrid, Spain. Electronic address: Carolina_Garcia-Canton@bat.com.
authors.forename: Carolina
authors.initials: C
authors.lastname: Garcia-Canton
authors.forename: Arturo
authors.initials: A
authors.lastname: Anadon
authors.forename: Clive
authors.initials: C
authors.lastname: Meredith
mesh_terms: D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D001980:Bronchi / Q000378:metabolism*; D002460:Cell Line; D004249:DNA Damage; D004847:Epithelial Cells / Q000378:metabolism*; D006657:Histones / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009152:Mutagenicity Tests / Q000379:methods; D020545:Respiratory Mucosa / Q000378:metabolism*; D012680:Sensitivity and Specificity
chemical_list: D015415:Biomarkers; C468783:H2AX protein, human; D006657:Histones
keywords: DNA damage; Double strand breaks; Genotoxicity; In vitro; γH2AX