Assessment of toxicity pathway responses in human-derived cell lines and the ability to distinguish adverse effects from adaptive changes is both an underpinning component of and key to the implementation of the TT21C strategy. High content analysis (HCA) provides one of the key approaches for developing mechanistic assays relevant to exploring and validating the network and circuitry of the toxicity pathways being assessed.
Over the past decade, the use of HCA has grown significantly combining the measurement of cellular biomarkers such as protein modifications, expression levels or cellular localisation, with changes in the morphology of the cell for target identification, pathway analysis or understanding mechanism of action. The main instrumentation used for our analysis comes from automated microscopic imaging systems, but also covers the use of flow cytometry systems. In both cases they integrate the assessment of phenotypic changes in whole cells performed on a multi-well platform together with automated data and image collection and analysis. These systems provide both increased throughput with simultaneous readout of multiple parameters performed at a single cell level across a large population of cells. The advantage is sensitive measures of the distribution of cell population behaviour in response to treatment, avoiding the reliance of using average cellular responses from a lysed population of cells. This importance of understanding not only averaged but individual responses has been shown by Batchelor et al. (Nat Rev Cancer. 2009 ; 9: 371–377) for a key signal transduction pathway related to one of our ongoing case studies on DNA damage.
While the different platforms have similarities they also provide complementary capabilities with one providing greater capacity in measurement of cell populations, the other capability in subcellular localisations; both important in developing our understanding and ability to model how changes in cell signal transduction dynamics occur across a dose response to a compound. This is key in the translation of data into a risk assessment decision related to the adaptive/adverse response.

Latest Presentation

Safety and Environmental Impact Assessment: An Industrial Perspective Biodynamics – Part 2 (In vitro Assays)

Development of an adverse outcome framework in the context of Nrf2 and oxidative stress responses to Quercetin and Curcumin

Latest Publication

Clewell RA, Sun B, Adeleye Y, Carmichael P, Efremenko A, McMullen PD, Pendse S, Trask OJ, White A, Andersen ME (2014) Profiling dose-dependent activation of p53-mediated signalling pathways by chemicals with distinct mechanisms of DNA damage, Toxicological Sciences, 142(1), 56-73

Seager AL, Shah UK, Brüsehafer K. Wills J, Manshian B, Chapman KE, Thomas AD, Scott AD, Doherty AT, Doak SH, Johnson GE, Jenkins GJ (2014) Recommendations, evaluation and validation of a semi-automated, fluorescent-based scoring protocol for micronucleus genotoxicity testing in human cell lines, Mutagenesis, 29, 155-164

Dr Sarah Cooper