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Evaluating drug-induced phenotypic and functional changes through multiple-readout assays

Quantitative high-throughput in vitro assays are an active area of investigation that enable assessment of phenotypic and functional changes using cell-based models from different sources.

One of the standard approaches for phenotypic assays is to evaluate the activity of biology modulators by establishing one single readout which has (but in most cases has not) been validated to be relevant for the therapeutic outcome. Recent reports have shown that modelling the activity of compounds in an in vitro system with one endpoint is often limited, if not misleading. This is particularly true for high complexity models such as those derived from the use of stem cells, primary cells and 3D cultures.

To tackle this issue, we employ high-content assay capabilities which are based on multiplexing and imaging readouts. Our experience with automated florescence microscopy spans more than two decades, and we can use additional state of the art technologies such as:

  • Confocal automated microscopy for screening
  • Live cell acquisition
  • Real time injection of modulators for fast kinetic readout
  • Image analysis and multivariate set mining.

We have applied high content imaging to a variety of applications, such as:

  • Primary neurons and astrocytes phenotypic screening (including mixed-cultures)
  • Immuno-oncology readouts such as proliferation, cluster formations and ADCC
  • Pharmacodynamics readouts (γH2AX for DNA damage as an example)
  • Fibrogenic-adipogenic differentiation of primary cells
  • Cardiomyocytes

In addition to automated fluorescence microscopy, we have strong skills in alternative cytometry readouts such as:

  • Plate based cytometry (via the TTP Acumen) which enables multiplexed fast data acquisition for higher throughput where a sub-cellular resolution is not required
  • FACS based cytometry for standard assays (such as cell cycle, apoptosis, differentiation, lymphocytes studies, T-cell activation, etc.) and also for ad hoc multiplexed readouts.

While evaluating the suite of tools that are required to run successful drug discovery programs, we always aim for multiple levels of readouts. This enables us to ensure we focus on mechanisms and modulators with the highest probability of success. The use of high content assays systems allows us to align the evaluation of primary end points with the simultaneous acquisition of additional features, suitable to dissect the experimental data for the most comprehensive interpretation of the results.

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