Abstract 2312: Using multidimensional real-time cell profiling approach to identify novel inhibitors of key targets and pathways involved in cancer maintenance and progression

Abstract

Abstract A high throughput cell-based phenotypic screening method was developed using real time cell analysis system which is based on impedance sensor technology. Using compounds with known mechanisms, we had previously demonstrated small molecule compounds produce characteristic time-dependent cell response profiles (TCRP) which could be indicative of mechanism of action. The cellular-basis for the anti-mitotic TCRP has been extensively validated using both anti-mitotic compounds and siRNAs targeting mitotic machineries (e.g. Eg5, PLK1 & Mad2). In this study, we used the TCRP approach to screen a 119,595 compound library for novel anti-cancer agents. The screening was performed at a final concentration of 17.8 µM using a human non small cell lung cancer line, H1993. The TCRP was monitored for 48 hours after compound treatment and the data generated was analyzed using in-house developed software to perform clustering analysis. For each screening set, a panel of compounds with known mechanisms of action were used as positive control and also served as reference for TCRP-based clustering to predict mechanisms of action. After multi-dose confirmation, 239 hits with IC50 values less than 8.9 µM were subjected to TCRP analysis. In comparison with reference TCRP patterns, we indentified 26 hits corresponding to tyrosine kinase inhibitors, 9 hits as DNA damaging agents, 125 hits as anti-mitotic agents, and 28 hits as cytostatic agents. Detailed characterization of 117 putative anti-mitotic compounds with traditional p-H3 based methods demonstrates 113 of them (96.6%) mediated mitotic arrest, thus demonstrating the robust predictive nature of the approach. We also have validated the putative tyrosine kinase inhibitors in in vitro kinase assays. TCRP-based phenotypic screening displays a novel drug screening approach with great potentials for discovering new pathway-modulating anticancer agents. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2312. doi:10.1158/1538-7445.AM2011-2312