Guava presented Simplicity Analysis Software and the Easycyte Plus System, which exploit the advantages of plate-based flow technology, at the Molecular Targets and Cancer Therapeutics Symposium.
These can speed-up drug discovery by extracting key findings from the complex data sets encountered in functional screening of siRNA knockdown assays.
Solid tumours comprise of genetically heterogeneous cell populations, the growth and survival of which depends on the complex interplay of distinct yet overlapping signalling networks.
A major challenge in developing a course of therapy is determining which signalling nodes to target for a specific malignancy.
Profiles from siRNA gene silencing are integral to mapping disease-specific signalling cascades and provide insight to key targets for therapeutic intervention.
Successful siRNA screening relies on optimising transfection and cell analysis systems capable of high content screening (HCS) at the single cell level, within overall populations (sample well) and across multiple data sets.
The presentation at the symposium described how the Guava Easycyte Plus System, with integrated Guava Simplicity Software, provides a platform for secondary target validation and compound screening.
Guava Technologies' flow cytometers overcome the limitations of inference-based measurements of transfection efficiency and protein knockdown through direct quantitiative analysis of populations at the single cell level.
The Simplicity Analysis Software allows research into multi-dimensional data sets through visualisation of user-defined parameters in the form of heat-maps.
Comparative results are displayed at the experiment level rather than on an individual well/sample basis.
The Easycyte Plus System was used in tandem with Simplicity Analysis Software to identify 23 agents with growth-restrictive properties, although significant variation across cell lines was observed.
Further targeted gene knockdown via siRNA confirmed the presence of both activators and inhibitors of Camptothecin-induced apoptosis as well as gene targets for growth arrest.
Screens for apoptosis and cell cycle and phospho-signalling intermediates defined compounds with mechanisms of action similar to and different from Camptothecin.
Cell-based assays for phenotype and function revealed many cooperative and antagonistic interactions between signalling intermediates, their respective cascades and cytoactive agents.
The acquired multiplex data set provides a more detailed view on the behaviour of each of the test compounds with respect to apoptotic induction, cell cycle progression and the signalling cascades that regulate these cellular responses to drug treatment.
