Multidimensional Drug Profiling By Automated Microscopy

Institute of Chemistry and Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Science (Impact Factor: 33.61). 11/2004; 306(5699):1194-8. DOI: 10.1126/science.1100709
Source: PubMed


We present a method for high-throughput cytological profiling by microscopy. Our system provides quantitative multidimensional measures of individual cell states over wide ranges of perturbations. We profile dose-dependent phenotypic effects of drugs in human cell culture with a titration-invariant similarity score (TISS). This method successfully categorized blinded drugs and suggested targets for drugs of uncertain mechanism. Multivariate single-cell analysis is a starting point for identifying relationships among drug effects at a systems level and a step toward phenotypic profiling at the single-cell level. Our methods will be useful for discovering the mechanism and predicting the toxicity of new drugs.

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Available from: Yan Feng, Jun 10, 2014
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    • "With few exceptions (Bodenmiller et al, 2012; Kleinstreuer et al, 2014), scalable methods to this end have been rather difficult to implement . In addition to transcriptome profiling (Lamb et al, 2006; Iorio et al, 2010), phenotypic profiling by cellular imaging has been deployed as a strategy for delineating a compound's mode of action by comparing drug-specific phenotypic responses (Perlman et al, 2004; Young et al, 2008; Gustafsdottir et al, 2013). "
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    • "Furthermore, integration of basic cell viability endpoints with gene expression profiling provide a useful source of biomarkers that predict sensitivity to cell-cycle arrest but poorly inform on optimal combination strategies or markers for other important cancer phenotypes such as apoptosis and invasion. Recent advances in fully automated brightfield and fluorescent microscopic acquisition platforms and associated image analysis algorithms have facilitated the integration of quantitative microscopic imaging of multiple endpoints upon both fixed and live-cells assays (Perlman et al., 2004; Yarrow et al., 2004; Tanaka et al., 2005; Caie et al., 2010). Screening beyond simplistic 2D monoculture assays is a necessary aim to target more relevant pathophysiological mechanisms and discover novel synergistic drug combination activity. "
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    • "plasma membrane permeability, cell morphology ). Similarly, automated fluorescence microscopy was used by Perlman et al. (2004) for multivariate single-cell analysis of HeLa cells to identify dose dependency and discriminate mechanisms of cytotoxicity for 100 drugs. They multiplexed a DNA stain with two of the following antibodyprobes per well: SC35, anillin; alpha-tubulin, actin, p38, ERK; p53, c-Fos, CREB and calmodulin. "
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