Validation of a human cell based high-throughput genotoxicity assay ‘Anthem’s Genotoxicity screen’ using ECVAM recommended lists of genotoxic and non-genotoxic chemicals

• Published in: Toxicology in vitro Vol. 28; no. 1; pp. 46 - 53
• Main Authors: Rajakrishna, Lakshmi, Krishnan Unni, Salini, Subbiah, Madhuri, Sadagopan, Sathish, Nair, Ayyappan R., Chandrappa, Ravindra, Sambasivam, Ganesh, Sukumaran, Sunil Kumar
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-02-2014
• Subjects: Colonic Neoplasms - metabolism; Drug Design; Drug Evaluation, Preclinical - methods; Drug-Related Side Effects and Adverse Reactions - diagnosis; ECVAM; GADD153; Genes, Reporter; HCT116 Cells; High-throughput; High-Throughput Screening Assays - methods; Human cell based; Humans; In vitro genotoxicity screen; Mutagenicity Tests - methods; Mutagens - toxicity; p21; p53; Reporter based screen; Sensitivity and Specificity; Time Factors; High-throughput; In vitro genotoxicity screen; GADD153; Reporter based screen; ECVAM; Human cell based; p21; p53
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2013.06.027

•A high throughput-enabled human cell based genotoxicity ‘Anthem’s Genotoxicity screen’ was developed.•Useful for rapid screening of genotoxins and reducing the number of animal studies.•Anthem’s Genotoxicity screen was evaluated using ECVAM recommended compounds.•The concordance of Anthem’s Genotoxicity screen with in vivo genotoxicity tests: 95.5%.•Sensitivity and specificity of Anthem’s Genotoxicity screen: 95.2% and 95.7%, respectively.
A novel high throughput-enabled human cell based screen, Anthem’s Genotoxicity screen, was developed to achieve higher specificity for predicting in vivo genotoxins by an in vitro method. The assay employs engineered human colon carcinoma cell line; HCT116 cells that are stably engineered with three promoter–reporter cassettes such that an increased reporter activity reflects the activation of associated signaling events in a human cell. The current study focuses on the evaluation of sensitivity and specificity of Anthem’s Genotoxicity screen using 62 compounds recommended by the European Centre for the Validation of Alternative Methods (ECVAM). The concordance of Anthem’s Genotoxicity screen with in vivo tests was 95.5% with sensitivity of 95.2% and specificity of 95.7%. Thus Anthem’s Genotoxicity screen, a high-throughput mechanism based genotox indicator test can be employed by a variety of industries for rapid screening and early detection of potential genotoxins.