NOV 12, 2014 8:00 AM PST

Hallmarks of cancer - Detect and quantify cell death signatures with high content imaging

Speaker
  • Oliver Kepp, PhD, INSERM staff scientist, Centre de Recherche des Cordoliers, Co-Director, Institut Gustave RoussyJayne A. Hesley, Applications Scientist, Cellular Imaging, Molecular Devices,
    Biography
      Oliver Kepp received his Ph.D. in 2006 from the Humboldt University of Berlin and the Max Planck Institute for Infection Biology in Berlin, Germany. He then joined the lab of Guido Kroemer in Paris, France as an Embo postdoc fellow. In 2012 he became an INSERM staff scientist (CR1 level). Currently he is co-directing the systems biology platform at the Gustave Roussy comprehensive cancer center where he investigates several aspects of immunogenic cell death, focusing on systems biology approaches.
       
      Jayne Hesley is an Applications Scientist for Cellular Imaging at Molecular Devices, LLC.  She has more than 6 years' experience developing cell-based applications using ImageXpress high content imaging systems and MetaXpress analysis software. Most recently her focus has been on utilizing stem-cell derived hepatocytes, cardiomyocytes and neurons for toxicity studies of cell viability, apoptosis, morphological changes, mitochondrial damage and autophagy. These studies include live-cell timelapse experiments as well as screening assays in 384 or 1536 well microplates. Prior to working in the imaging field, Jayne developed colorimetric and fluorescence assays, including TR-FRET and fluorescence polarization, for high-throughput screening on multi-mode readers. She has a B.S. in Animal Science and M.S. in Biology.

    Abstract

    Dr. Oliver Kepp will be presenting on:

    Immunogenic cell death fingerprinting utilizing a high-throughput screening approach

    The strategy of immunogenic cell death fingerprinting has been designed for the purpose of identifying novel immunogenic anticancer agents in drug re-profiling approaches. Following a general high-throughput screening (HTS) approach (Kepp et al., Nat Rev Drug Discov., 2011) chemical compound libraries have been analyzed by robot-assisted automated microscopy using fluorescent biosensors that were generated for detecting immunogenic cell death signatures (CRT exposure, HMGB1 and ATP release (Zitvogel, Kepp, Kroemer, Cell, 2010)). Using this approach we recently identified compounds from the group of cardiac glycosides, of which some have reached FDA approval for the treatment of heart insufficiency, to be capable of inducing all known hallmarks of immunogenic cell death. The emission of immunogenic cell death factors, triggered by the primary hit compounds, was further validated by low throughput assays and the immunogenic potential of few leads was confirmed in vivo by means of immunocompetent mice and retrospective clinical studies. This successful HTS approach using fluorescent biosensors underlines the capacity of our microscopy-based platform to detect signatures of immunogenic cell death in a large number of different treatment conditions.


    Jayne Hesley will be presenting on:

    High-content imaging for accurate quantification of autophagy in neuronal cells

    Autophagy is a process in which unwanted cellular components, from aged organelles to faulty proteins, are sequestered into double-membrane compartments called autophagosomes.  Autophagy is key to maintaining cell homeostasis, as disruption in the process is implicated for conditions such as neurodegeneration, aging, and cancer.

    Autophagosomes can be visualized and characterized even in heterogeneous cell populations using readily available reagents, automated imaging and high-content analysis of live or fixed cells.  The ImageXpress® Micro XLS system makes it possible to study a large number of treatments with replicates, since it enables acquisition of high resolution, high magnification images at multiple fluorescent wavelengths from cell-based assays run in 384 well microplates. Dose-response curves from human iPSC-derived neurons or PC-12 cells were generated from multiple data outputs, allowing discrimination between simple cytotoxicity and specific effects on cellular function.  Accurate co-localization measurements of mitochondria or lysosomes with autophagosomes provide a deeper understanding of the mechanisms of autophagic stimulation or inhibition.

     

     


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