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SELECTBIO Conferences Exosomes & Liquid Biopsies Europe 2018

Exosomes & Liquid Biopsies Europe 2018 Agenda


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Wednesday, 24 October 2018

08:00

Conference Registration, Materials Pick-Up, Morning Coffee and Breakfast Pastries


Session Title: Conference Opening Session Framing the Key Topics and Opportunities in Liquid Biopsy

09:00

Dominique PV de KleijnConference Chair

Conference Chairman Welcome and Opening Remarks
Dominique PV de Kleijn, Professor Experimental Vascular Surgery, Professor Netherlands Heart Institute, University Medical Center Utrecht, The Netherlands, Netherlands

09:15

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.

10:00

Alain ThierryKeynote Presentation

Circulating DNA as a Next Generation Diagnostic Tool
Alain Thierry, Director of Research, Institut de Recherche en Cancérologie de Montpellier-INSERM, France

Circulating tumor DNA (ctDNA) holds promises in tumor burden monitoring or malignancies early detection. Based on crucial observation on the structure and origin of ctDNA we developed a multiparametric method for specifically analyze ctDNA. This technology enables the first clinical validation of the analysis of circulating DNA in oncology, in a prospective blinded multicentric study for the detection of KRAS and BRAF mutations from colorectal patient plasma samples. Aims of our research are now focusing on various aspects of the potential of ctDNA: (i), detecting the emergence of the mutations following targeted therapy; (ii), developing the Intplex test for the multimarker quantitative analysis of ctDNA; (iii), studying the follow up of CRC patients; (iv), the prognostic power of ctDNA and (v) the screening power of ctDNA.

10:45

Morning Break and Networking in the Exhibit Hall

11:15

Klaus PantelKeynote Presentation

Liquid Biopsy: The New Diagnostic Concept in Oncology
Klaus Pantel, Director, University Of Hamburg, Germany

The analysis of circulating tumor cells (CTCs) in blood may provide clinically relevant information as “liquid biopsy” (Alix-Panabieres & Pantel, Nature Rev Cancer 2014) and provide new insights into tumor biology (Uhr & Pantel, PNAS, 2011, Lu et al., Cancer Cell, 2011; LeBleu et al., Nat. Cell Biol., 2014; Werner et al., Cancer Discovery, 2015). Besides CTCs the molecular analysis of ctDNA provides important complementary information as “liquid biopsy” (Pantel & Alix-Panabieres, Cancer Res., 2013; Pantel et al., Nature Med. 2013; Heitzer et al., Genome Med. 2013; Schwarzenbach et al., Nature Rev. Clin. Oncol., 2014, Joosse & Pantel, Cancer Cell 2015; Bardelli & Pantel, Cancer Cell 2017). Moreover, circulating microRNAs, extracellular vesicles and tumor-educated platelets are also interesting new members of the “Liquid Biopsy family” with potential clinical relevance in the future. In particular exosomes have gained great interest because they act as biomarkers with important functional properties for tumor progression. E.g., the integrin composition of exosomes can determine the organ site of metastases (Hoshino, Pantel, Lyden et al., Nature, 2015) and microRNAs in exosomes can impact the biology of the recipient cells (Anfossi, Bababayan, Pantel, Calin, Nature. Rev. Clin. Oncol. 2018). Detection of these miRNAs can contribute to early detection of cancer (Meng et al., Oncotarget, 2016). Liquid biopsy analyses with validated platforms provides reliable information on prognosis and may serve to identify therapeutic targets or mechanisms of resistance on metastatic cells. Metastatic cells might have unique characteristics that can differ from the bulk of cancer cells in the primary tumor currently used for stratification of patients to systemic therapy. Moreover, monitoring of blood samples before, during and after systemic therapy (e.g., chemotherapy, hormonal therapy, antibody therapy) might provide unique information for the future clinical management of the individual cancer patient and might serve as surrogate marker for response to therapy. In conclusion, liquid biopsies can be used to improve the management of individual cancer patients and contribute to the vision of personalized medicine (Alix-Panabieres & Pantel, Cancer Discovery, 2016; Bardelli & Pantel, Cancer Cell 2017). Validation of liquid biopsy assays is essential and is currently being performed by the EU/IMI consortium CANCER-ID (www.cancer-id.eu).

12:00

Catherine Alix-PanabièresKeynote Presentation

Functional Analyses of Circulating Tumor Cells in Cancer Patients
Catherine Alix-Panabières, Director of the Laboratory of Rare Human Circulating Tumor Cells, University Medical Centre of Montpellier, France

Circulating tumor cells (CTCs) in blood are promising new biomarkers potentially useful for prognostic prediction and monitoring of therapies in patients with solid tumors including colon cancer. Moreover, CTC research opens a new avenue for understanding the biology of metastasis in cancer patients. However, an in-depth investigation of CTCs is hampered by the very low number of these cells, especially in the blood of colorectal cancer patients. Thus, the establishment of cell cultures and permanent cell lines from CTCs has become the most challenging task over the past year.  We described, for the first time, the establishment of cell cultures and a permanent cell line from CTCs of one colon cancer patient (Cayrefourcq et al. Cancer Res. 2015). The cell line designated CTC-MCC-41 is in culture for more than three years and has been characterized at the genome, transcriptome, proteome and secretome levels. This thorough analysis showed that CTC-MCC-41 cells resemble characteristics of the original tumor cells in the colon cancer patient and display a stable phenotype characterized by an intermediate epithelial/mesenchymal phenotype, stem-cell like properties and an osteomimetic signature indicating a bone marrow origin. Functional studies showed that CTC-MCC-41 cells induced rapidly in vitro endothelial cell tube formation and in vivo tumors after xenografting in immunodeficient mice.  More recently, we defined the molecular portrait of these metastasis-competent CTCs (Alix-Panabières et al. Clin Chem. 2017). These results highlight that CTC-MCC-41 line display a very specific transcription program completely different than those of the primary and metastatic colon cancer cell lines. Interestingly, among the 1,624 transcripts exclusively up-regulated in CTC-MCC-41 samples, key genes related to energy metabolism, DNA repair and stemness genes were observed. Such data may supply insights for the discovery of new biomarkers to identify the most aggressive CTC sub-populations and for the development of new drugs to inhibit metastasis-initiator CTCs in colon cancer.  Moreover, the development of new immunotherapeutic strategies is of utmost importance. Antibodies against proteins that block the immune response of T-cells such as PDL1 have been approved for treatment of cancer patients after showing remarkable long-term remissions in subsets of patients. It is now important to develop predictive biomarkers to identify patients with the highest benefit from these therapies. In 2015, we could show for the first time that PD-L1 is heterogeneously expressed on CTCs from metastatic breast cancer patients (Mazel et al. Mol Oncol 2015). Further functional analysis of this interesting subset of CTCs might reveal special immunosuppressive properties.

12:45

Networking Lunch in the Exhibit Hall -- Meet the Exhibitors and View Posters


Session Title: Circulating Biomarkers for Cancer

14:00

Minetta LiuKeynote Presentation

Circulating Biomarkers in the Management of Solid Tumors
Minetta Liu, Associate Professor and Chair, Oncology Research, Mayo Clinic, United States of America

The real-time, serial detection of tumor specific molecular alterations will facilitate improved clinical outcomes through individualized treatment approaches. The advent of technologies that allow for the isolation and characterization of circulating tumor cells (CTCs) and cell free nucleic acids (cfNA) in the peripheral blood has led to great interest in the “liquid biopsy”. These assays offer a less invasive, potentially more cost effective tool to assess prognosis and response to treatment, and they may facilitate the early diagnosis of recurrence or primary malignancy itself. The challenges of validating these molecular biomarkers and the need to provide solutions to promote rapid translation into clinical practice will be discussed.

14:45

Liquid Biopsy: Screening for Single Nucleotide Variants and Indels From Cell-Free Tumoral DNA
Yves Rozenholc, Professor, University of Paris Descartes, France

Analyzing circulating free DNA from blood sample of patient known to have a cancer may help to build a precise picture of the tumoral mutations driving the carcinogenesis process, which can be used as input of a personalized treatment and offers the opportunity to realize a precise follow-up of patient under treatment. However, detecting single-nucleotide variations and insertions/deletions in circulating tumor DNA is challenging because of their low allele frequency. Based on quantification of error rate at each base position [position error rate (PER)], I will present a screening tool called PlasmaMutationDetector, which allows to identify mutations as the result of a statistical test comparing minor-allele frequency to measured PER at each base position. The method is based on a non-optimized commercial kit and allows actually the screening of about 20000 mutations (SNV and INDEL) potentially occurring along several oncogenes. It shows excellent sensibility and specificity and allows detection when minimal mutated allele frequencies is as low as 0.3% for single-nucleotide variations and 0.1% for insertions/deletions. During this presentation, to help adaptation of this method to other oncogenes and/or other NGS techniques, I will present the main technical steps, which allow to deal with multiplicity, hotspot and PER estimation.

15:15

Glioblastoma Extracellular Vesicles Hijack Multiple Microglial Functions to Promote Tumor Growth
Marieke Broekman, Neurosurgeon, University Medical Center Utrecht and Massachusetts General Hospital/Harvard Medical School, United States of America

Extracellular vesicles (EVs) have been shown play a role in glioblastoma biology. Contrary to all prior studies, we will present data on EV mediated modification of microglia and macrophages in vivo.

15:45

Afternoon Coffee Break and Networking in the Exhibit Hall

16:15

Clinical Applications and Limitations of Plasma cfDNA
Malgorzata Ilona Srebniak, Clinical Cytogeneticist, Erasmus Medical Centre, Netherlands

The clinical applications and limitations of cytogenomic analysis in plasma cfDNA will be shown based on the NIPT in prenatal samples and tumor cfDNA studies in hematological malignancy and in solid tumors.

16:45

Diagnostic Leukapheresis (DLA) to Improve CTC-Based Liquid Biopsies
Nikolas Stoecklein, Professor, Experimental Surgical Oncology, Heinrich Heine University Düsseldorf, 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 resulted in an increase in CTC detection frequency and an escalation of the median CTC numbers. We hope that DLA, or similar approaches derived thereof, will advance the clinical utility of CTC-based liquid biopsies.

17:15

Daniel ChiuKeynote Presentation

The Isolation and Analysis of Rare Cells, Exosomes, and Circulating Nucleic Acids For Liquid Biopsy
Daniel Chiu, A. Bruce Montgomery Professor of Chemistry, University of Washington, United States of America

This presentation will describe a rare-cell isolation instrument we call eDAR (ensemble decision aliquot ranking) for the automated enrichment and analysis of rare cells, a nanofluidic technology for the high-sensitivity and high-purity isolation of exosomes, and a digital-nucleic-acid detection platform based on our SD (self digitization) chip. I will outline the workings of these techniques, describe their performance, and discuss the clinical questions we are addressing to demonstrate the utility of liquid biopsy in precision medicine.

18:00

Networking Reception in the Exhibit Hall with Beer and Wine. Engage with the Exhibitors, View Posters in a Relaxed Setting at the Marriott Rotterdam

19:00

Close of Day 1 of the Conference.

Thursday, 25 October 2018

07:30

Morning Coffee, Breakfast Pastries and Networking

08:00

Fred KramerKeynote Presentation

Breakfast Lecture: Multiplex Real-Time PCR Assays that Assess the Abundance of Rare Somatic Mutations Associated with Cancer
Fred Kramer, Professor of Microbiology, Biochemistry & Molecular Genetics, New Jersey Medical School Rutgers University, United States of America

“SuperSelective” PCR primers, due to their unique design, exponentially amplify rare mutant DNA fragments present in a sample, while ignoring abundant related wild-type DNA fragments, even if the only difference between the mutant and the wild-type is a single-nucleotide polymorphism.  Different target mutations whose presence suggests the same therapeutic action can be detected with molecular beacons bearing the same colored fluorophore.  Differently colored molecular beacons are simultaneously present in these assays to determine which therapeutic action should be taken and which therapeutic action should be discontinued.  These multiplex, homogeneous assays are sensitive, rapid, inexpensive, utilize existing PCR instruments, and can be carried out frequently during the course of treatment utilizing non-invasive liquid biopsy samples.


Session Title: Deploying Exosomes, CTCs, and Circulating Nucleic Acids in Cancer, Cardiovascular Disease and CNS Diseases

09:00

Dominique PV de KleijnKeynote Presentation

Vesicles For Ischemic Heart Disease
Dominique PV de Kleijn, Professor Experimental Vascular Surgery, Professor Netherlands Heart Institute, University Medical Center Utrecht, The Netherlands, Netherlands

Cardiovascular Disease (CVD) is with the cardiovascular events of Ischemic Heart Disease and Stroke, the number 1 and 2 cause of death in the world and expect to increase especially in Asia. Ischemic heart disease (IHD) comprises 3 entities: stable coronary artery disease (SCAD), unstable angina (UA) and myocardial infarction (MI). Because IHD is associated with an increased risk of adverse clinical events such as heart failure and death, early recognition of IHD is of utmost importance. However, to diagnosis IHD is challenging, as many patients present with atypical symptoms. It is known that women have a different symptom sensation than men. Troponins are the main diagnostic tool for detection of MI. Blood biomarkers for SCAD (typically causing stable angina) and UA, however, are not available. These diagnoses frequently require hospital visits/admissions for time-consuming and costly (non)invasive tests. We use plasma extracellular vesicle protein content of vesicles from plasma subfractions as an accurate source for early diagnosis of SCAD and UA. Diagnosis of ischemic heart disease as well as normalization and characterization of extracellular vesicles in plasma subfractions will be discussed.

09:45

Molecular and Functional Characterization of CTCs in Non-Small Cell Lung Cancer (NSCLC)
Francoise Farace, Head of the Translational Laboratory of Rare Circulating Cells, INSERM Gustave Roussy Institute, France

The presentation will focus on the liquid biopsy potential of CTCs to analyze resistance mutations to tyrosine kinase inhibitors in ALK-rearranged and EGFR-mutant patients. Our work on NSCLC CDX (CTC-derived xenograft) will be presented and discussed.

10:15

Accuracy, Reproducibility and Consequences of Bias in Quantitative Small RNA-seq For Circulating Cell-free RNA Profiling
Maria Giraldez, Researcher, University of Michigan, United States of America

Small RNA-seq is increasingly being used for profiling of small RNAs, including cell-free RNA such as microRNAs present in plasma and other biofluids. In practice, small RNAseq can involve many different methods for library preparation, with distinct protocols and technical variations that could cause substantial differences in results, yet these have not been fully and systematically studied. We report here the results of a study using common references (synthetic RNA pools of defined composition, as well as plasma-derived RNA) to evaluate the accuracy, reproducibility and bias of small RNA-seq library preparation for five distinct protocols and across nine different laboratories. We observed protocol-specific and sequence-specific bias, which was ameliorated using adapters with randomized end-nucleotides for ligation. Despite this technical bias, relative quantification using small RNA-seq was remarkably accurate and reproducible, even across multiple laboratories using different methods. These results provide a foundation for reproducible, cross-laboratory small RNAseq studies of cell-free, circulating RNA in a variety of clinical contexts.

10:45

Morning Coffee Break and Networking

11:15

How Clinical Biobanks Support Precision Medicine: From Standardized Preprocessing of Liquid Biopsies to Treatment Guidance
Jens Habermann, Scientific Director, Surgical Center for Translational Oncology, University of Lübeck and University Hospital Schleswig-Holstein, Germany

CTCs and Exosomes harbor an enormous potential for precision medicine. However, clinical implementation requires standardization. This is in contrast to current praxis, which is still hampered by major inter-study heterogeneity. This talk will address (i) current challenges and solutions for standardized CTC / Exosome assessment and (ii) how clinical biobanks can support CTC / Exosome research for precision medicine, e.g. in colorectal cancer.

11:45

Exploiting Extracellular Nucleic Acids From Body Fluids 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 body fluids as extracellular mRNAs, lncRNAs and microRNAs, ready to exploited. I will discuss various workflows for RNA sequencing of body fluid derived RNA, including probe-based target capture as a sensitive RNA sequencing workflow to study thousands of mRNA and lncRNA genes in cell-free RNA from patients’ plasma and urine. 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 body fluid 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 exRNAQC study.

12:15

Networking Lunch in the Exhibit Hall

14:00

Lucia LanguinoKeynote Presentation

Exosomal Integrins: Biomarkers for Liquid Biopsy
Lucia Languino, Professor of Cancer Biology, Thomas Jefferson University, United States of America

Cancer Extracellular Vesicles (EVs) found in blood are heterogeneous and show unique protein compositions. Our analysis shows that a prostate cancer EV subset, designated exosomes, carries a unique repertoire of integrins, which are surface receptors for extracellular matrix.  Using iodixanol gradients, immunoblotting and size measurements we show that integrins are differentially expressed in cancer exosomes. Our recent studies demonstrate that exosomes from prostate cancer cells contribute to horizontal propagation of unique integrin-associated invasive phenotypes.  We also show that exosomal integrins are transferred to cells in the tumor microenvironment and mediate response to therapy. This presentation will discuss the role of exosomal integrins in cancer progression and therapy response, and their potential use as clinical biomarkers for non-invasive diagnosis of cancer.

14:45

Liquid Biopsy in Colon Cancer
Pierre Laurent, Professor, Universite Paris Descartes, France

RAS mutations are currently sought for in tumor samples, which takes a median of almost 3 weeks in western European countries. This creates problems in clinical situations that require urgent treatment and for inclusion in therapeutic trials that need RAS status for randomization. Analysis of circulating tumor DNA might help to shorten the time required to determine RAS mutational status prior to anti-EGFR antibody therapy for metastatic colorectal cancer. Here we compared plasma versus tissue RAS analysis in a large prospective multicenter cohort.

15:15

Afternoon Coffee Break

15:45

Droplet-Based Digital PCR Procedures and Optimized NGS to Track Circulating Tumor DNA in Liquid Biopsies: Application to 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. In particular, droplet based digital PCR has allowed to reach unprecedented sensitivity and accuracy for rare sequences detection. Complementarity of these technologies will be presented through several retrospective and prospective studies. Moreover, we will present their application for the detection of circulating tumor DNA and its potential use for cancer patient treatment management.

16:15

Approaches For Validation of Liquid Biopsy Testing of Circulating Tumour DNA
Alison Devonshire, Science Leader, LGC Limited, United Kingdom

The typically low concentration of ctDNA in cancer patients’ samples makes validation of analytical sensitivity a critical element in the assessment of a diagnostic test. The potential for real-time monitoring of tumor dynamics through liquid biopsy testing also necessitates a systematic evaluation of quantitative accuracy.  This presentation will discuss approaches to validate the performance of targeted methods for cancer mutations, including pre-analytical controls, and the development of reference materials and reference measurement procedures, which also support longer-term comparability and standardization of patient testing.

16:45

Takahiro OchiyaKeynote Presentation

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

17:30

Panel Discussion: Emerging Themes and Trends in Liquid Biopsy Development

18:15

Close of Day 2 of the Conference.

Friday, 26 October 2018

08:00

Morning Coffee, Breakfast Pastries and Networking


Session Title: Exosomes, Microvesicles and Extracellular Vesicles (EVs) -- Research, Diagnostic and Therapeutic Opportunities

09:00

Sai Kiang LimKeynote Presentation

Exploiting the Unique Protein Distribution in Different Plasma EV Types For Biomarker Discovery
Sai Kiang Lim, Research Director, Institute of Medical Biology, A*STAR, Singapore

Intercellular communication via secreted lipid membrane vesicles or extracellular vesicles (EVs) is an emerging research area.  Many cell types are known to secrete EVs and these EVs are heterogenous in size, density, composition and biogenesis pathway.  EVs in the size range of 100-200 nm are among the most intensely studied EVs, and are frequently referred to as exosomes which are EVs derived from endosomes.  Recently, we demonstrated by pulse chase experiments that Mesenchymal Stem Cell (MSC) exosomes can be identified by their high affinity for cholera toxin B chain (CTB) which binds GM1 gangliosides.  MSC was also observed to secrete at least two other EV types that can be distinguished by their affinity for lipid-binding proteins namely, AnnexinV (AV) that binds phosphatidylserine and Shiga toxin B chain (STB) that binds globotriasosylceramide.  These three EV types have different protein and RNA cargo.  Several other cell types were also observed to produce CTB-, AV- and ST-binding EVs (CTB-EVs, AV-EVs and ST-EVs).    Evaluation to date suggest that these different EV types are unique, and carry different protein and RNA cargos.  We have since demonstrated that plasma also have CTB-EVs, AV-EVs and ST-EVs.  Hence plasma biomarkers can now be analyzed in four permutations: plasma, plasma CTB-EVs plasma AV-EVs or plasma ST-EVs.  The distribution of some plasma proteins in these four permutations is different among individuals, and could be associated with diseases.   As such, plasma CTB-EV and AV-EV could be sources of novel biomarkers or novel permutations of known biomarkers.  Here I will describe using this approach to discover three biomarkers in plasma, plasma CTB-EV and plasma AV-EV of 28-32 weeks pregnant women that could predict development of PE at an average of seven weeks later.  This study was done using a prospective biobank of 843 low risk patients with a 2.1% PE risk.  At 100% sensitivity, the three biomarkers have a combined AUC of 0.96, 78.6% specificity and a PPV of 9.9%.  I will also present the discovery of ovarian cancer biomarker candidates using similar approaches.

09:45

Giovanni CamussiKeynote Presentation

Cancer-derived Extracellular Vesicles and Tumor Microenvironment
Giovanni Camussi, Professor, University of Torino, Italy

This presentation will cover the role of extracellular vesicles in cell-to-cell communication. It will also address the correlation between the molecular composition of extracellular vesicles and their biological activity. The  ability of tumor and cancer stem cell derived extracellular vesicles to induce epigenetic changes in tumor micro-environmental cells, to inhibit  the differentiation process of DCs and to favor the formation of a pre-metastatic niche will also be covered.

10:30

Samir EL-AndaloussiKeynote Presentation

Purification and Characterization of Engineered EVs
Samir EL-Andaloussi, Assistant Professor, Karolinska Institutet, Sweden

Extracellular vesicles (EVs) have emerged as important mediators of intercellular communication due to their ability to transfer bioactive lipids, proteins and different species of RNA into cells. Thus, EVs hold therapeutic potential in their own right and can additionally be harnessed for the delivery of macromolecular drugs.  This presentation will cover the developments in producing, purifying and characterizing EVs. It will also cover the different strategies to engineer producer cells in order to generate EVs harboring selected RNAs or proteins for treatment of inflammatory diseases, with focus on neuro inflammation.

11:15

Short Coffee Break

11:30

Title to be Confirmed.
Esther Nolte-‘t Hoen, Assistant Professor, Department of Biochemistry & Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Netherlands

12:00

Lunch and Networking in the Exhibit Hall

13:00

Paul RobbinsKeynote Presentation

Title to be Confirmed.
Paul Robbins, Professor, The Scripps Research Institute, United States of America

13:45

Niek DekkerKeynote Presentation

Engineered Extracellular Vesicles for CRISPR/Cas9 Gene Editing
Niek Dekker, Associate Director, Discovery Biology, AstraZeneca, Sweden

Extracellular vesicles (EVs) represent an exciting opportunity as biological delivery vehicles for therapeutic cargo with excellent safety, low intrinsic immunogenicity, cell-specific tropism and biological delivery efficiency. There are multiple approaches for the introduction of protein and RNA cargo into EVs, including physical, chemical and cell engineering. Application of CRISPR/Cas9 requires the functional delivery of the ribonucleoprotein (RNP) complex of Cas9 enzyme and guide RNA. Engineering of the production cell line can be used to introduce cargo into the EVs, and examples for the introduction of fluorescent and luminescent proteins will be disclosed. Uptake in recipient cells can be monitored using high content imaging to provide insight in kinetics and localization. Functional delivery can be analyzed using several assays for the gene editing event (eg Cel1 and TIDE). The use of EVs for delivery of CRISPR/Cas9 has a range of exciting applications for ex vivo manipulation of primary cells and in vivo editing of genes in diseased tissue.

14:30

Challenges in the Detection, Isolation and Quantification of Extracellular Vesicles and Exosomes
Alain Brisson, Professor Emeritus, University of Bordeaux, France

The emerging field of extracellular vesicle (EV) and exosome research faces a notorious lack of reliable characterization methods. This is mainly due to the small size of EV/exosomes (mostly from 30 to 500 nm) and the heterogeneity of their suspensions. This presentation will illustrate the advantages of novel methods allowing to image, phenotype, quantify and isolate EVs in complex fluids.

15:00

Circulating Microvesicles – Biomarkers and Effectors and Cardiovascular Health and Disease
Felix Jansen, Leader of the Microvesicle Research Group, Department for Cardiology, Pulmology and Angiology, University Hospital Bonn, Germany

My talk will summarize the diagnostic potential and current obstacles in using circulating MVs as biomarkers in CVD. Furthermore, I will highlight the regulatory role of MV in the progression of atherosclerosis.

15:30

Afternoon Coffee Break

16:00

Exosome-derived Epigenetic Biomarkers For Saliva Diagnostics
Christa Noehammer, Senior Scientist, Austrian Institute of Technology, Austria

The aim of our research activities at AIT, the Austrian Institute of Technology, is to define reliable biomarkers suitable for early and non-invasive disease diagnosis and prognosis. To this end we have been establishing and optimizing a whole range of multiplex capability technologies (e.g. microarrays, quantitative PCR, Luminex bead technology) to meet the special demands and challenges of diagnostic biomarker discovery - and validation in body fluids. Using this specific technology expertise we e.g. successfully discovered autoantibody- as well as DNA methylation -based diagnostic marker panels for the big 4 cancer entities (breast, colon, prostate, lung) in serum or plasma. Based on these success stories and the evident advantages of saliva as a diagnostic matrix our recent special interest is to go for saliva diagnostics and to evaluate saliva for its suitability for circulating biomarker-based diagnostics.  Along these lines we will report on the evaluation of different commercially available strategies for isolation of exosomes from human serum and saliva. We will further present data from genome-wide microRNA – as well as DNA-methylation profiling in salivary - and serum-derived exosomes which will also include our experiences when comparing the performance of high-density microarrays and next generation sequencing for miRNA profiling. Last but not least we will report on first salivary and plasma exosome-derived epigenetic biomarkers for type 2 diabetes diagnosis.

16:30

Title to be Confirmed.
Marcin Jurga, R&D Director, The Cell Factory bvba, Netherlands


Add to Calendar ▼2018-10-24 00:00:002018-10-26 00:00:00Europe/LondonExosomes and Liquid Biopsies Europe 2018Exosomes and Liquid Biopsies Europe 2018 in Rotterdam, The NetherlandsRotterdam, The NetherlandsSELECTBIOenquiries@selectbiosciences.com