Wednesday, 30 October 2019

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Title to be Confirmed.
Balaji Panchapakesan, Professor, Department of Mechanical Engineering, Worcester Polytechnic Institute (WPI), United States of America

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Leon TerstappenKeynote Presentation

Circulating Tumor Cells to Guide Treatment of Cancer Patients
Leon Terstappen, Chair Medical CellBiophysics, MIRA Research Institute for Biomedical Technology and Technical Medicine, University of Twente, Netherlands

The presence of Circulating Tumor Cells (CTCs) enumerated with the CellSearch system is associated with relatively poor survival and their reduction after the first weeks of therapy indicates a response to therapy.  Present research focus is on the ability to extract treatment relevant information from the proteins, RNA and DNA in these CTC.

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Title to be Confirmed.
Felix Jansen, Leader of the Microvesicle Research Group, Department for Cardiology, Pulmology and Angiology, University Hospital Bonn, Germany

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Considerations and Quality Controls When Analyzing Cell-Free Tumor DNA
Anders Ståhlberg, Associate Professor, Sahlgrenska Cancer Center, University of Gothenburg, Sweden

Analysis of circulating cell-free tumor DNA (ctDNA) in liquid biopsies offers new means for early cancer diagnostics, real-time monitoring of treatment efficiency and detection of relapse. Despite its potential use ctDNA remains challenging to detect and to quantify as it represents only a small fraction of all cell-free DNA. We have developed an ultrasensitive sequencing technology, SiMSen-Seq, that allows allele frequencies < 0.1% to be detected. SiMSen-Seq is simple to perform, flexible in multiplexing and requires minimal DNA input. Here, we discuss important considerations for ctDNA detection in plasma, including all experimental steps from sampling to data interpretation. We will also discuss how the use of high fidelity enzymes reduce error rates in barcoded NGS. The use of quality control assays enables the development of robust and standardized workflows that facilitate the implementation of ctDNA analysis into clinical routine.

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Exosomal-miRNA Signatures in Circulation
Michiel Pegtel, Associate Professor, Cancer Center Amsterdam, VU University Medical Center, Netherlands

The peripheral blood is rich in cell-free microRNAs (miRNA) that are derived from both living and dying cells. A proportion of these miRNAs are protected from RNAses within vesicles including apoptotic bodies, microvesicles and exosomes. In vitro, exosomes can transport miRNAs to recipient cells, which may also occur in vivo although the physiological relevance and molecular mechanisms of cell-cell transfer of miRNAs has not been established. In this presentation I will discuss our clinical studies concerning exosomal miRNAs as cancer biomarkers for therapy response evaluation and the potential of exosomes as therapeutic delivery vehicles.

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Graham PockleyKeynote Presentation

The Theranostic Potential of Membrane Hsp70 in Cancer: Development of a Liquid Biopsy and an Approach for Isolating Circulating Tumour Cells (CTCs)
Graham Pockley, Director and Professor, John van Geest Cancer Research Centre, Nottingham Trent University, CEO, multimmune GmbH, United Kingdom

A cell surface bound form of the 70 kDa heat shock (stress) protein Hsp70 is selectively and widely expressed on the plasma membranes of many tumour entities. This presentation will describe the development of a liquid biopsy for identifying patients bearing membrane Hsp70-expressing tumours, the current status of an NK cell-based therapeutic for the treatment of such tumours, and, separately, the development of an approach for isolating circulating tumour cells (CTCs) based on their expression of membrane Hsp70.

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Biomarker-based Strategy to Solve Resistance to Immunotherapy
RJ Tesi, CEO/CMO, Inmune Bio Inc, United States of America

In cancer, resistance to immunotherapy can be managed differently than resistance to chemotherapy.  Resistance to immunotherapy does not mean abandoning the immunotherapy, but adding a second immunotherapy that overcomes the immunologic resistance mechanism.  Combination immunotherapy should be precision medicine based of rational, biologically based and measurable biomarkers to select the subset of patients who will be sensitive to therapy.  Preclinical data on resistance immune checkpoint inhibitors and trastuzumab resistance will be used as case studies.

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Standardized Analysis of Extracellular Vesicles in Liquid Biopsies
An Hendrix, Professor, Ghent University, Belgium

The identification of extracellular vesicle (EV)-associated biomarkers is challenging owing to the complexity of liquid biopsies. We 1) performed quality control studies to identify the impact of (pre-) analytical variables on biomarker identification, 2) developed reference materials to ensure standardized EV measurements, and 3) created EV-TRACK to stimulate researchers to put experimental guidelines into practice. This combined expertise boosted the identification of bacterial EV in the systemic circulation of patients with intestinal barrier dysfunction.

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Title to be Confirmed.
Irina Nazarenko, Exosome and Tumor Biology Group Leader, University of Freiburg, Germany

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High Blood Volume Approaches to Improve CTC-based Liquid Biopsies
Nikolas Stoecklein, Professor, University of Dusseldorf, Germany

The direct access to systemically spread cancer cells is the true potential of CTC-based liquid biopsies. However, the major obstacle to implement CTC-based liquid biopsies into clinical routine applications is the extreme low concentration of CTCs and the minimal amount of investigated blood in standard CTC-tests. To tackle this problem, we introduced Diagnostic Leukapheresis (DLA), which increased significantly CTC detection frequency and resulted in an escalation of CTC numbers. We hope that DLA, or similar approaches derived thereof, will advance the clinical utility of CTC-based liquid biopsies. Besides DLA, I will review some new alternative approaches also aiming to increase the investigated blood volume in my talk.

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Patrizia Paterlini-BrechotKeynote Presentation

Clinical Applications of Circulating Tumor Cells to Predictive Oncology
Patrizia Paterlini-Brechot, Professor of Oncology/Molecular Biology, University Paris Descartes, France

Circulating Tumor Cells (CTC) and Circulating Tumor Microemboli (CTM) are Circulating Rare Cells which herald tumor invasion and are expected to provide an opportunity to improve the management of cancer patients and help to detect the most aggressive invasive cancers before other diagnostic approaches like imaging. We report on the clinical applications of CTC in predictive oncology and more specifically on the ISET® (Isolation by SizE of Tumor/Trophoblastic Cells) open system for marker-independent isolation of tumor cells from blood which allows to reliably diagnose and count CTC.

Independent published data from several international research teams have demonstrated the prognostic relevance of CTC detection by ISET® in patients with melanomas, as well as lung, colorectal, liver,  pancreatic, head and neck, and ovarian cancers. Furthermore, the utility of theranostic characterization of CTC by ISET® has been demonstrated for non-small-cell lung cancers, colorectal cancers and melanomas.

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Tracking Circulating DNA for Cancer Patient Follow-Up
Valérie Taly, CNRS Research Director, Group leader Translational Research and Microfluidics, University Paris Descartes, France

Tracking of circulating cell-free nucleic acids in body effluents has been greatly facilitated by recent technological developments including droplet-based digital PCR and optimized NGS. These technics now allow to reach unprecedented sensitivity and precision for rare sequences detection and quantification. Strategies allowing analysis of several genetic and epigenetic cancer-specific alterations have will be presented for the detection of tumor DNA in patient samples. Moreover to demonstrate the pertinence of such procedures to overcome clinical oncology challenges, the results of several clinical studies will be presented. In particular, we will illustrate the pertinence of these approaches to monitor circulating tumor DNA in plasma (so-called liquid biopsy) for the follow-up of patients with localized or advanced cancers.

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CTC Genetic Diversity through Single-Cell Analysis and CDX Development
Francoise Farace, Head of the Translational Laboratory of Rare Circulating Cells, INSERM Gustave Roussy Institute, France

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Leveraging the Genome-Wide Cell-Free DNA Fragmentation in Patients With Cancer
Florent Mouliere, Assistant Professor, Cancer Center Amsterdam, VUMC Amsterdam, Netherlands

Liquid biopsies, and circulating tumor DNA (ctDNA) in particular, are used to non-invasively detect the presence of cancer cells. ctDNA is commonly detected though the presence of genetic alterations in its sequence. Since the majority of ctDNA in circulation is non-mutated, leveraging this untapped resource of tumor information makes sense for improving cancer detection, notably in early stage and brain cancers. Here I will show how the genome-wide fragmentation of cell-free DNA could be leveraged to enhance the detection of ctDNA.

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Title to be Confirmed.
Jens Habermann, Scientific Director, Surgical Center for Translational Oncology, University of Lübeck and University Hospital Schleswig-Holstein, Germany

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Characterization and Quantification of Extracellular Vesicles and Exosomes
Alain Brisson, Professor Emeritus, University of Bordeaux, France

Extracellular vesicles (EV) from body fluids or conditioned media are heterogeneous mixtures containing not only bona fide EV differing by their cell or membrane origin, size, phenotype or cargo content, but also various types of non-vesicular particles and other components, either isolated or aggregated. Given the diversity of biological functions and biomedical applications borne by EV, there is an urgent need for efficient and reliable methods of EV charaterization, quantification and isolation, to improve our understanding on EV and fully exploit their potential.

This presentation will illustrate the power of immuno-gold cryo-electron microscopy combined with magnetic isolation and flow cytometry for providing a detailed and quantitative characterization of EV mixtures.

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Saliva, Exosomes and Type 2 Diabetes Diagnostics
Christa Noehammer, Senior Scientist, Austrian Institute of Technology, Austria

Saliva is a readily and even within short time intervals repeatedly available body fluid, which can be obtained via non-invasive, painless collection. In addition, saliva represents a basically unlimited sample matrix and further offers the advantage of not requiring any special skills for collection. Saliva contains a broad range of relevant molecule classes, such as DNA, messenger RNA, micro RNA, lipids, proteins and antibodies, which all can be used as potential disease/health biomarkers. Along these lines Saliva diagnostics is an emerging field of molecular diagnostics, not least because scientific findings increasingly show that measuring and quantifying salivary biomolecules can be used not only to detect local diseases of mouth and throat but also to diagnose systemic diseases. Due to these opportunities and advantages a recent special interest of the Molecular Diagnostics research group at the AIT Austrian Institute of Technology is to evaluate saliva for its suitability for circulating biomarker-based diagnostics.  Along these lines we will show proof of concept studies for autoantibody- and DNA-methylation based salivary diagnostics and report on the evaluation of different commercially available strategies for isolation of exosomes from human serum and saliva. We will further present data from comparative profiling studies in salivary - and serum-derived exosomes including targeted protein-, genome-wide microRNA – as well as DNA-methylation profiling. Last but not least we will report on results of a research project where we are looking for salivary and plasma exosome-derived epigenetic biomarkers for early type 2 diabetes diagnosis.

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Exploiting RNA In Liquid Biopsies For Precision Medicine Purposes
Jo Vandesompele, Professor, Ghent University and CSO, Biogazelle, Belgium

In contrast to general belief, a substantial part of the human transcriptome is abundantly present in the blood and other biofluids as extracellular messenger RNA, long non-coding RNAs and various small RNAs, ready to exploited. I will discuss various workflows for RNA sequencing of biofluid derived RNA, including probe-based target capture and unbiased total RNA library prep as sensitive RNA sequencing workflow to study thousands of mRNA and lncRNA genes in cell-free RNA from patients’ plasma and other biofluids. Apart from RNA abundance profiling, this type of data can also be used to detect structural RNA variants, such as somatic mutations, fusion genes and RNA editing events, all known to play an important role in disease, including cancer. The resulting RNA profiles can be deconvoluted to enumerate the cells, tissues and organs that contribute to the extracellular RNA. Human biofluid RNA sequencing enables liquid biopsy guided precision oncology, such as therapy stratification, treatment response monitoring and early detection of relapse. I will also discuss the pre-analytical jungle of RNA targeted liquid biopsies and need for standardization, as part of the ongoing extracellular RNA quality control study. I will end with the first insights of the Human Biofluid RNA Atlas, in which we have deeply probed into the extracellular transcriptome of 22 human biofluids, providing a solid foundation for exploiting biofluids for diagnostic purposes.

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Paul RobbinsKeynote Presentation

Title to be Confirmed.
Paul Robbins, Professor, Department of Biochemistry, Molecular Biology, and Biophysics, and the Institute on the Biology of Aging and Metabolism, University of Minnesota Medical School, United States of America

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Isolation and Multiplex Analysis of Tumor Cells and Tumor DNA From Body Fluids on a Chip
Lorena Diéguez, Leader of the Medical Devices Research Group, INL- International Iberian Nanotechnology Laboratory, Portugal

Early dissemination of cancer is difficult to detect by traditional imaging and pathological methods. While the presence of cancer material in body fluids is well known, current techniques for the isolation, analysis and characterization of these biomarkers are not efficient enough to be fully applied in clinical routine.
In recent years, microfluidics has been widely reported for efficient isolation of CTCs and free nucleic acids. Moreover, microfluidics and microdroplets offer miniaturization, automation, high-throughput, and intrinsic reproducibility. Despite fluorescence has been the main detection technique integrated within microfluidics and microdroplets, SERS allows higher detection limits and offers much more information than fluorescence, as the Raman spectrum is the fingerprint of the analyte under study and allows multiplex detection.

In this talk, we present our work for isolation and multiplex analysis of cancer biomarkers from body fluids based on microfluidics, microdroplets and SERS. The present approach combines the device engineering for size isolation, with nanotechnology engineering and optical detection. The development of this platform encounters a remarkable advance towards personalized medicine, especially in the field of cancer research.

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Liquid Biopsy -- Exosomal microRNA Biomarker Signatures in Clinical Diagnostics
Michael Pfaffl, Professor, Technical University of Munich, Germany

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Takahiro OchiyaKeynote Presentation

Title to be Confirmed.
Takahiro Ochiya, Chief, National Cancer Center Research Institute Japan, Japan

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Distribution of Non-Coding RNAs Over EV and Other RNA Carriers in Plasma
Esther Nolte-‘t Hoen, Assistant Professor, Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Netherlands

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Oxford Nanoimaging (ONI)The Nanoimager: Resolving EVs One Molecule at a Time
Mark Harmon, Field Applications Specialist, Oxford Nanoimaging (ONI)

ONI crafts desktop-compatible microscopes with miniaturized Nobel Prize winning technology capable of detecting individual molecules using fluorescence.  This presentation will introduce our capabilities with accompanying images.

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Klaus PantelKeynote Presentation

Title to be Confirmed.
Klaus Pantel, Director, University Of Hamburg, Germany

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Catherine Alix-PanabièresKeynote Presentation

Title to be Confirmed.
Catherine Alix-Panabières, Director of the Laboratory of Rare Human Circulating Tumor Cells, University Medical Centre of Montpellier, France

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Title to be Confirmed.
Sara Colombetti, Global Head Oncology Discovery Pharmacology, pharma Research and Early Development (pRED), Roche Innovation Center Zürich, Switzerland

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Approaching Alzheimer’s Disease as an Immunological Disease: Role of Biomarkers
CJ Barnum, Director of Neuroscience, INmune Bio, Inc., United States of America

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Zero Hour Liquid Biopsies from Donor Kidneys
Bas WM van Balkom, Assistant Professor, University Medical Center Utrecht, Netherlands

Kidney transplantation is the preferred treatment for end-stage kidney disease, and donor kidney shortage urges proper donor-recipient matching. Zero-hour biopsies provide predictive value for short- and long-term transplantation outcome, but are invasive and may not reflect the entire organ. Alternative, more representative methods to predict transplantation outcome are required. We hypothesized that preservation fluid contains proteins and exosomes could serve as biomarkers to predict post-transplantation graft function. We explored their presence and biomarkers potential and confirmed that secreted products in this fluid can be used to predict post-transplantation kidney function.

Thursday, 31 October 2019

00:00

Title to be Confirmed.
Lucia Languino, Professor of Cancer Biology, Thomas Jefferson University, United States of America