07:30 | Morning Coffee, Tea, Breakfast Pastries and Networking |
| Session Title: High Content and Phenotypic Screening |
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08:00 | High-Content Screening for Adverse Cytologic Effects Peter O'Brien, Head of Clinical Pathology, University College Dublin, Ireland
The first, highly-effective, high-content-screening (HCS) for adverse cytologic effect was reported a decase ago (Arch Toxicol 2006, 80, 580). Success was attributed to simultaneous measurement of multiple “cytobiomarkers”, use of human cells that can metabolise drugs, 72 h exposure for slow toxicants, exposure to wide-ranging concentrations, and normalizing toxic to efficacious concentration. Many HCS studies now support this approach as necessary in predictive toxicology, as does review of literature since the ?rst cytotoxicity assay was reported 100 years ago. A subset of the original toxicants was reanalyzed using the original HCS confirming high sensitivity and speci?city across locations, technologies, sta?, laboratories, and time. A protocol is demonstrated for operational validation of the HCS within labs to document pro?ciency and quality management. |
08:30 | Using Patient-Specific Induced Pluripotent Stem Cells to Model Retinal Diseases Alice Pébay, Associate Professor, Principal Research Fellow, University of Melbourne & Centre for Eye Research Australia, Australia Maciej Daniszewski, Researcher, University of Melbourne, Australia
Age-related macular degeneration (AMD), primary open-angle glaucoma (POAG) are leading ageing neurodegenerative diseases of the eye. Although phenotypically distinct, their respective pathophysiologies do share similarities. They are chronic, progressive, and lead to the degeneration of neurons key to the function of the retina or optic nerve. If no treatment is found, the direct and indirect costs associated with these conditions will drastically increase. But Despite enormous research efforts, treatment options are still limited, with no current definitive treatment for AMD and POAG. This paucity of treatment options can be attributed to a fundamental absence in the knowledge around what causes these conditions and how they progress. We use of patient-specific induced pluripotent stem cells (iPSCs) to obtain the key cells involved in these diseases. To increase power of analysis, we use automation of cell culture to allow large-scale modelling of AMD and POAG. Those methods will be discussed. Ultimately, we aim to improve our molecular understanding of those neurodegenerative diseases, and ultimately facilitate preclinical trials, and translatable outcomes. |
09:00 | More Content from High-Content Screening: Analyzing Specimen in 3D Alexander Schreiner, Team Leader Biological Applications, PerkinElmer
When developing a screening assay one of the most critical aspects is the choice of cellular model including the decision to choose between 2D or 3D approaches. Multicellular “oids” (tumoroids, spheroids, organoids) bear the potential to better predict drug candidate effects during preclinical screening. However, 3D models require more effort and compared to 2D cultures, 3-dimensional assays tend to be more complex across all steps of the workflow. Reliably generating sufficiently large numbers of uniform spheroids for screening and imaging of 3D volumes at high quality are among the challenges. Using 3D spheroids and cysts as examples, we will show experimental workflows, explaining how to effectively grow these models using ULA coated U-bottom plates or low concentration gels. The careful selection of dyes and clearing strategies can improve the image quality while targeted imaging of spheroids helps to significantly shorten imaging times and to minimize the data volume. Nevertheless, being able to effectively analyze spheroids is still one of the main bottlenecks. Dedicated high-content software tools can help users to, speed up 3D image acquisition by targeted imaging of objects of interest, better understand the spatial context of your 3D cell models through different visualization methods, measure volume and morphology changes in 3D to characterize your specimen in detail and export 3D movies to share and publish your results. |
09:30 | Exploring in vitro Vibrational Microspectroscopy for High Content Analysis Hugh Byrne, Professor, Dublin Institute of Technology, Ireland
The potential of vibrational spectroscopy, but infrared absorption and Raman scattering, in the fields of biological and biomedical applications has been well demonstrated. The techniques provide molecular based profiles of the biomolecular content, in a label free, non-destructive manner, which, in the microscopic mode, can have subcellular resolution. Multivariate statistical analysis is commonly employed to extract high content information concerning cellular metabolism and response pathways. The presentation provides an over view of efforts to advance the application of vibrational spectroscopy for high content cellular analysis, using examples of exposure to model nanoparticles and chemotherapeutic agents. This progress is discussed in the light of emerging technologies in the field of spectroscopic microscopy. |
10:00 | Coffee Break and Networking |
10:30 | Collaborative Phenotyping at King’s: HipSci and the Stem Cell Hotel Davide Danovi, Director, Cell Phenotyping Platform, Kings College London, United Kingdom
We work in the framework of the Human Induced Pluripotent Stem Cells Initiative (HipSci) project, funded by the Wellcome Trust and MRC. We provide a dedicated laboratory space for collaborative cell phenotyping to study how intrinsic and extrinsic signals impact on human cells to develop assays for disease modeling and drug discovery and to identify new disease mechanisms. |
11:00 | High-Content Screening of Complex Biological Models Ben Haworth, Cellular Imaging Application Specialist, GE Life Sciences
Exploring how the IN Cell Analyzer Platforms delivers image quality and
speed in high-content Analysis workflows. We will look at how the
systems can be used to optimize assays using both 3D and live models. |
11:30 | Enabling Early Stage Muscle Drug Discovery with MyoScreen™, a High-throughput High-content Phenotypic Screening Platform Deploying Micropatterned Human Primary Skeletal Myotubes Joanne Young, Senior Scientist, CYTOO SA
CYTOO has recently developed a disease relevant phenotypic muscle drug discovery platform to fully leverage the characterization of compounds at an early stage of the drug discovery pipeline. MyoScreen™ deploys imaging and image-analysis methodology along with a micropatterned plate system that allows the precise control of myotube length and the formation of highly aligned myotubes. In this seminar, I will discuss the robust performance of the MyoScreen™ platform and show how it exploits the improved architecture and maturity of micropatterned myotubes, which are superior to standard 2D culture, to quantitatively analyze drug effects on proliferation, hypertrophy/atrophy, acetylcholine receptors, as well as providing a screening index of contractile force. I will also describe actual use of MyoScreen™ in a primary screen with secondary hit validation, focusing on two intriguing hits that were identified to enhance myotube function by distinct mechanisms of action. |
12:00 | | Keynote Presentation Multidimensional Fluorescence Imaging For High Content Analysis and Preclinical Applications Paul French, Professor, Department of Physics and Photonics Group, Imperial College London, United Kingdom
We are developing multidimensional fluorescence imaging technology with a particular emphasis on fluorescence lifetime imaging (FLIM) across the scales to contrast different molecular species and to map variations in the local fluorophore molecular environment, particularly due to Forster resonant energy transfer (FRET) in order to assay protein interactions or read out genetically expressed FRET biosensors. For high content analysis (HCA) we are implementing automated time-gated FLIM for multiwell plate assays of protein interactions or cellular metabolism. We have applied FLIM FRET HCA to study signalling and disease mechanisms in 2-D and 3-D cell-based assays, including the intracellular measurement of KD to quantify protein interactions. For in vivo preclinical studies, we are combining optical projection tomography (OPT) with FLIM, particularly applied to live zebrafish - from larvae up to adults - and are developing FLIM endoscopy to read out FRET biosensors in murine disease models. We aim to implement our multidimensional fluorescence imaging technology using open source software tools for instrument control, data acquisition, analysis and management and to provide lists of equipment components to enable other users to replicate our instrumentation for application to their own biological questions. Our open source FLIM analysis software, FLIMfit which provides rapid global fitting capabilities and is available as an OMERO client and our FLIM HCA and OPT instrumentation are controlled by µManager. |
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12:30 | Robust & Efficient Tools for Pluripotent Stem Cell Research Wing Chang, R&D Director, STEMCELL Technologies, United Kingdom
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13:00 | Networking Lunch -- Meet the Exhibitors and View Posters |
| Session Title: Antibodies in Drug Discovery and Emerging Areas |
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13:30 | Nanobodies Against Influenza and Their Applications in Preparing For Future Pandemics Simon Hufton, Section Head Molecular Immunology, National Institute for Biological Standard and Control (NIBSC), United Kingdom
Both seasonal and pandemic influenza continue to represent a formidable
public health challenge. The constant evolution of new strains means the
rapid availability of highly specific neutralising monoclonal
antibodies are of considerable interest. Nanobodies are emerging as next
generation antibodies with a number of well documented advantages which
make them highly amenable to applications in the infectious disease
area. This presentation will describe the isolation and characterization
of nanobodies specific for key influenza subtypes including ‘swine flu’
A(H1N1), highly pathogenic avian influenza A(H5N1), highly pathogenic
avian influenza A(H7N9), A(H3N2) and B lineage influenza viruses. The
use of yeast display for mapping their epitopes on hemagglutinin will
also be presented as will their potential applications in preparing for
future influenza pandemics. |
14:00 | Exploiting the Multiple Functions of FcRn for Therapy E. Sally Ward, Professor, Texas A&M University Health Science Center, United States of America
The central role of FcRn in regulating IgG persistence and transport
provides opportunities for targeting this receptor in multiple different
diagnostic and therapeutic situations. The engineering of IgGs with
higher affinity for FcRn can be used to produce antibodies with longer
in vivo half-lives, but only if the pH dependence of the IgG-FcRn
interaction is retained. Conversely, engineered IgGs with increased
affinity for FcRn at both acidic and near neutral pH act as potent
inhibitors of FcRn. Consequently, such antibodies (‘Abdegs’, for
antibodies that enhance IgG degradation) can lower the levels of
endogenous IgG. Recent studies in our laboratory have also resulted in
the generation of engineered Fc-fusions that selectively clear
antigen-specific antibodies (‘Seldegs’, for selective degradation).
Recent developments related to the modulation of IgG dynamics will be
presented. We have demonstrated that the loss of expression of FcRn in
tumor cells results in increased intracellular albumin accumulation and
tumor growth in mouse models. These observations have implications for
tumor immunotherapy and indicate a novel function for FcRn as a
metabolic regulator. |
14:30 | | Keynote Presentation New Ways For Human Antibodies - From Intracellular Applications to Switchable Affinity Stefan Dubel, Full Professor and Director, Technische Universität Braunschweig, Germany
While the generation and optimization of human antibodies by phage display today allows to design fine specificity and kinetic parameters in detail, and CAR-T-cells, ADCs, bi-specific antibodies and many others creative derivatives of the original IgG molecule are approved or in clinical studies, the antibody molecule still offers many opportunities for new and innovative applications. The access of intracellular antigens with biologicals would add a huge target space to therapeutic antibodies. We used intra-bodies in the first protein knock down mouse to demonstrate the in vivo function of this principle, but also analysed many strategies to insert antibodies from the outside of the cell. On another end, using cyclic mutants of human calmodulin as an allosteric effector module, antigen binding affinity of different antibodies could be regulated. This allosteric effect was demonstrated for five different scFv fragments under physiological conditions on living cells without the need of pH or ion concentration changes. |
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15:15 | | Keynote Presentation Yeast as a Compound-Target Pair Discovery Tool Stephen Helliwell, Senior Investigator, Novartis Institutes for BioMedical Research, Switzerland
I will briefly summarize our efforts at Novartis to identify yeast bioactive LMWs and identify their targets using uHTS screens and chemogenomic profiling – known as HIP HOP. |
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16:00 | Overcoming Antigen-Mediated Antibody Clearance Tristan Vaughan, Vice President, R&D, Antibody Discovery and Protein Engineering, MedImmune Ltd, United Kingdom
An antigen can act as a “sink” for antibody clearance if the antibody binds to an antigen which is rapidly eliminated. High levels of antibody clearance can impede and often halt the development of a novel therapeutic due to an impossibly large dose that would be required to overcome the sink for the drug to bring any patient benefit. Here, an antibody engineering solution to rapid clearance will be described and a case study presented.
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16:30 | Enhanced Interrogation of the B Cell Repertoire For Antibody Discovery Win-Yan Chan, Research Scientist, UCB, United Kingdom
An overview of the automated, high through-put antibody discovery platform at UCB and our single-cell recovery technology for antibody discovery. Using a sophisticated screening cascade at an early stage of antibody discovery, we are able to find high quality antibodies that will go on to make the best therapeutic candidates. |
17:00 | Close of Day 2 of the Conference |