Fridtjof Lund-Johansen
Head of flow cytometry core facility, Oslo University Hospital, Norway
When
10th-11th Oct 2019
Registration from 8am
Where
London, United Kingdom
London Heathrow Marriott Hotel
Discover the latest technological developments and applications of flow cytometry on cellular analysis and in cancer diagnosis and treatment. Focus is given to advances in flow cell and sheath fluid design, the development of new fluorophores, and next-generation detection systems. The applications session covers the analysis of cell surface antigens, reviewing cell health and death status, cell sorting techniques and drug development in oncology.
As part of the Research & Technology Series: Immuno-oncology you will have full access to 100+ presentations across five meetings and an exceptional interdisciplinary networking and learning experience.
THURSDAY 10TH OCTOBER 2019 – STRATEGY & TECHNOLOGY
Keynote Address: Semi-supervised data clustering and machine learning for leukemia diagnosis
RICHARD SCHEUERMANN, Director, Adjunct Professor, J. Craig Venter Institute
The current approach for identifying diagnostic cell populations from cytometry data in clinical labs is based on manual gating analysis, which is subjective and labor-intensive, especially with higher-dimensional complex datasets. Over the last several years, our group and others have developed a suite of computational tools and machine learning for the processing and analysis of cytometry data. To illustrate the potential use of these methods in a clinical diagnostic setting, we will present the results of a pilot project to optimize and apply a selected set of computational and machine learning methods for the automated identification of chronic lymphocytic leukemia (CLL) cells in patient samples
Keynote Address: Imaging Flow Cytometry – combining statistically powerful flow cytometry with information rich microscopy
ANNE WILSON, Director of the Flow Cytometry Facility, University of Lausanne (UNIL) Switzerland)
Conventional flow cytometry is a statistically powerful technology used to analyze antigen expression in populations of cells at the single cell level. It however provides little information about cell morphology, interactions between cells and localization of antigens or molecules on or within the cells, or, co-localization of molecules to cytoplasmic or nuclear structures. Imaging Flow cytometry provides all this detailed information and more at the single cell level on large numbers of cells, combining the best features of both conventional flow cytometry and microscopy. This presentation provides examples of the power and diversity of this technology.
Presentation
Morning Refreshments / Poster Presentations
Seeing more by targeting less – development of probes, prodrugs and provenance
RACHEL ERRINGTON, Chair and Principle Investigator of the Tissue MicroEnvironment Group Division of Cancer and Genetics, School of Medicine, Cardiff University
The development of both fluorescent probes is an important endeavour in cancer research useful for the drug discovery pipeline. Our simple working hypothesis is that the cellular uptake and subcellular localisation of molecules can provide important information on the status of a cell. Core to the Deep-RedAnthraQuinone (DRAQ) probe family is that the spectral-window for both excitation and emission sits in the red (>600nm) thereby making the probe suitable for studies on single cells (2D) through to thick tissue models (3D ie spheroids and organoids). Our key undertaking has been to design molecules that allow us to tune their capacity to label the nucleus versus other cellular compartments providing a greater range of signal readouts, for tracking behaviour and the emergent properties of populations
The Development of a Multicolour Antibody Panel for Mouse Bone Marrow Stromal Cells – Challenges and Breakthroughs
JANE SRIVASTAVA, Flow Cytometry Facility Manager, South Kensington Campus, Imperial College London
Why is this panel important?
• Why we chose these specific markers and fluorophores
• Some tips for the challenges of working with a large multicolour antibody panel with rare cells
• Final panel considerations and preliminary data
• Future work
Lunch
Implementation of Multi-Parameter Flow Cytometry Assays in Clinical Trials
VILMA DECMAN, Associate Director Flow Cytometry, Bristol-Myers Squibb
Flow cytometry is a powerful technique in the research, drug development and clinic as it allows for the examination of multiple targets on various cell subsets from a limited sample size. Its ability to identify and enumerate different cellular markers and track them across time and treatment aids in understanding of disease pathology, toxicological assessment of new drugs and their efficacy. This talk will concentrate on implementation of multi-parameter flow cytometry in clinical trials including:
• Understanding of biomarker needs in development of high-dimensional flow cytometry assays
• Design and optimization of panels; potential pitfalls
• Validation of assay/standardization
• Sample logistics
• New technology platforms
An in vivo approach to the early photoacoustic detection of infectious diseases and blood conditions
VLADIMIR ZHAROV, Professor, Director, University of Arkansas for Medical Sciences, Arkansas Nanomedicine Center
60 MINUTE INTERACTIVE WORKSHOP
Afternoon Refreshments / Poster Presentations
Standardizing technical practise across labs – developing an automated shared resource library
CHRIS GROVES, Senior Manager, MedImmune
Data-driven cytometry
RYAN BRINKMAN, Professor, Medical Genetics, University of British Columbia Distinguished Scientist, BC Cancer CEO, Cytapex Bioinformatics Inc.
Manual analysis of flow cytometry data is subjective and time consuming. Automated algorithms have reached a level of maturity that enables them to match and, in many cases, exceed the results produced by human experts. The current state-of-the-art will be reviewed though example applications of these algorithms in automating the entire data analysis pipeline. Examples will include automated quality checking at the event and sample level, rapid, robust and reproducible automated gating and biomarker discovery algorithms. The utility of these algorithms will be shown through their application on patient data for basic research, clinical trials and drug discovery.
Networking Drinks Reception
FRIDAY 11TH OCTOBER 2019 – CELLULAR ANALYSIS & APPLICATIONS IN ONCOLOGY
Keynote Address: From stem cell to blood cell: flow cytometry of the differentiation pathway
FRANK PREIJERS, Radboud University Medical Center, Nijmegen, The Netherlands
Flow cytometry (FCM) is increasingly used in clinical laboratories for diagnosis of various hematological disease. The rapid and sensitive multi-parameter detection renders FCM to a powerful tool to distinguish malignant cells from normal. Pattern recognition of fluorescence expression levels of conjugates and combinations in the various cells activation stages is hereby essential. However, before aberrancies can be established, the normal hematopoiesis must be studied. Bone marrow contains all differentiation stages of hematopoiesis. The maturation can be monitored by changes in immunophenotype, identified by a unique combination of MoAbs.
Each antigen is expressed by an appropriate expression pattern during the maturation. Frequently, neoplastic cells possess aberrant immunophenotypes. More or less antigens and antigen expression on different levels can be found in these leukemic cells. This implies that knowledge of the normal cell maturation patterns facilitates recognition of malignant cells. We studied the maturation pathways from stem cells to myeloid, monocytic and erytroid lineage cells using a 10-color NaviosTM (Beckman Coulter) and KaluzaTM analysis software.
Identifying new forms of cell death – using flow cytometry to explore cell fate
GARY WARNES Flow Cytometry Core Facility Manager, Blizard Institute, Barts and London School of Medicine & Dentistry, Queen Mary London University
• RIP3 and Caspase-3 intracellular labelling with a fixable live/dead probe allows the flow cytometric detection of necroptosis (RIP3high+ve/Caspase-3-ve) , apoptosis (RIP3-ve/Caspase-3+ve) and RIP1-dependent apoptosis (RIP3+ve/Caspase-3+ve)
• Further labelling with LC3B allows the detection of autophagic cells
• Additional labelling with H2AX and PARP allows further identification of DNA damage, hyper-action of PARP and parthanatos
• Incidences of RCD were modulated by the use of zVAD and Necrostain-1
• Autophagy was shown to protect cells by reducing DNA damage
• Use of MitoTracker and violet live cell caspase probe allows the identification of necroptosis and apoptosis in unfixed cells
• Additional use of fixable probes for mitochondrial function and Reactive Oxygen Species (ROS) extends the number of identifiable cell death populations to 500
Morning Refreshments / Poster Presentations
Starting up clinical trials with TCR-modified T cells in solid cancers
DAVID BAKER Senior Research Scientist, AstraZeneca
• The high-throughput flow cytometry capability at AZ and examples of how this has been used.
• Applications of high-throughput flow cytometry in immuno-oncology projects (assays such as T-cell proliferation, activation, tumour killing assays).
Applications of flow cytometry in early phase oncology trials
STEPHANIE TRAUB,Cancer Research UK
The presentation will outline the use of flow cytometry in early phase clinical development with relevant examples of commonly encountered challenges and suggested solutions.
Lunch
Developing a GUCY2C-Targeted Adoptive T Cell Therapy
FRIDTJOF LUND-JOHANSEN, Head of flow cytometry core facility, Oslo University Hospital, Norway
Proteomics is still a small discipline where the research front is driven by a few laboratories with unrestricted access to mass spectrometry (MS). With MAP, we put the power of proteomics into the hands of flow cytometry (FCM) users. Bead-based arrays with a multiplexing capacity up to 5000 provide the convenience and throughput needed to take proteomics to the next level. The challenge lies in changing the mindset that keeps the western blot on top of the list of the most popular antibody applications. The FCM version is called Western-MAP and yields results for thousands of antibodies in parallel. In Native-MAP, proteins are separated by size exclusion chromatography for large-scale analysis of protein complexes. Thus, MAP turns the flow cytometer into a high throughput proteomics machine.
Analysing t cells for neo-antigen specificity
PIA KVISTBORG (Reserved), Junior Group Leader, NKI Amsterdam
Chair’s Closing Remarks / Conference Close
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Poster presentation sessions will take place in breaks and alongside the other breakout sessions of the conference. Your presentation will be displayed in a dedicated area, with the other accepted posters from industry and academic presenters.
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