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SELECTBIO Conferences Circulating Biomarkers World Congress 2020

Circulating Biomarkers World Congress 2020 Agenda

Co-Located Conference Agendas

Circulating Biomarkers World Congress 2020 | EV-based Diagnostics, Delivery & Therapeutics | 

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Monday, 17 February 2020


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

Session Title: Conference Opening Session -- Emerging Themes in Circulating Biomarkers and Exosomes/EVs


Michael GranerConference Chair

Welcome and Introduction to the Conference
Michael Graner, Associate Professor, Dept of Neurosurgery, University of Colorado Anschutz School of Medicine, United States of America


Steve SoperKeynote Presentation

Integrated Microfluidic Systems for the Efficient Isolation of Circulating Leukemia Cells and Circulating Plasma Cells
Steve Soper, Foundation Distinguished Professor, Director, Center of BioModular Multi-scale System for Precision Medicine, The University of Kansas, Adjunct Professor, Ulsan National Institute of Science & Technology, United States of America

Liquid biopsies are generating great interest within the biomedical community due to the simplicity for securing important biomarkers to manage complex diseases. We are developing a suite of microfluidic devices that can process whole blood directly and engineered to efficiently search for a variety of disease-associated liquid biopsy markers and their subsequent molecular analysis. One microfluidic device can isolate targets with recovery >90% and high purity (>80%) to enable downstream analysis of the particular biomarker without requiring single cell picking. We have also developed a microfluidic device for imaging single cells. The aforementioned microfluidic devices can be interfaced to a fluidic motherboard to allow for full process automation, including cell selection from blood and then, immunophenotyping and/or FISH of the isolated cells. In this presentation, information will be shared on the operational parameters of these devices for the selection of liquid biopsy markers, and the downstream molecular information that can be garnered from the isolated markers in acute myeloid / acute lymphoblastic leukemia (circulating leukemia cells) and multiple myeloma (circulating plasma cells). The attractive nature of using liquid biopsy markers for these two diseases is that it circumvents the need for a patient to undergo a painful bone marrow biopsy. Information will be provided as to the use of these liquid biopsy markers to monitor relapse from minimum residual disease, and stage patients for directing therapy.


Daniel ChiuKeynote Presentation

New Flow-Based Technologies for the High-Sensitivity and High-Resolution Analysis of CTCs and Exosomes
Daniel Chiu, A. Bruce Montgomery Professor of Chemistry, University of Washington, United States of America

This presentation will describe two flow-based technologies we developed for the analysis of single rare cells and individual extracellular vesicles. The first flow platform is a rare-cell isolation instrument we call eDAR (ensemble decision aliquot ranking), which is capable of operating in a rapid sequential sorting mode for isolating single rare cells with exceptionally high sensitivity and purity. The second flow instrument is a single-vesicle sorter, capable of high-sensitivity analysis of individual extracellular vesicles. I will outline the workings of these new tools, describe their performance, and discuss the clinical questions we are addressing with these next-generation technologies.


Min YuKeynote Presentation

Circulating Tumor Cells Inform Mechanism of Breast Cancer Metastasis
Min Yu, Assistant Professor, University of Southern California, United States of America

Circulating tumor cells (CTCs) are expected to contain metastasis-initiating cells that can shed light on the mechanisms of cancer metastasis. However, due to limited patient-derived material, the metastatic capability of CTCs has yet to be proved. Using our recently established patient-derived CTC lines, we found that different patient CTC lines demonstrated distinct metastatic tissue tropisms in immunodeficient mice and identified associated pathways to specific organs via RNA-seq and ATAC-seq analysis.


Morning Coffee Break and Networking in the Exhibit Hall


Biofluid Biomarkers for the Brain
Kendall Van Keuren-Jensen, Professor and Deputy Director, Translational Genomics Research Institute, United States of America

We will describe some of our efforts in using exRNAs as diagnostics for central nervous system injury and disease.


Jennifer JonesKeynote Presentation

Liquid Biopsies Using EVs: Promise and Peril on the Frontiers of a New Field
Jennifer Jones, NIH Stadtman Investigator, Head of Transnational Nanobiology, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, United States of America


Randolph CortelingKeynote Presentation

An Exosome-based Drug Delivery Platform Derived From an Immortalized Human Neural Stem Cell (hNSC) Line
Randolph Corteling, Head of Research, ReNeuron Ltd., United Kingdom

To ensure the scale required for clinical research and commercialisation producer cell immortalisation and clonal isolation is a practical strategy to produce consistent, functionally bioactive exosomes for use as therapeutic agents. Immortalisation ensures production stability and reduces the need for equivalence testing.

CTX0E03 is a conditionally immortalised human neural stem cell line that has been manufactured to clinical-grade (GMP) standards, using a 3-tier banking strategy and is currently in Phase IIb clinical evaluation for disability after stroke. Using the conditioned media produced during GMP manufacture, we have shown that CTX0E03 is an abundant producer of exosomes that can be readily isolated and purified at scale. The CTX cell line can also be rapidly and efficiently modified to direct the expression of a variety of cargoes within the secreted EV population, whilst maintaining the key immortalised stem cell characteristics of the parental cell line.  The natural tissue tropism of CTX-derived exosomes can then be exploited to deliver loaded cargoes to target cells.


Susanne GabrielssonKeynote Presentation

Optimizing Dendritic Cell-Derived Exosomes For Cancer Immunotherapy
Susanne Gabrielsson, Professor, Division of Immunology and Allergy, Department of Medicine Solna, Karolinska Institutet, Sweden

Peptide loaded exosomes are promising cancer treatment vehicles, however, low T cell responses in human clinical trials indicate a need to further understand exosome-induced immunity. We previously demonstrated that antigen-loaded exosomes carry whole protein antigens and require B cells for induction of antigen-specific T cells. I will discuss our latest data where we investigated the need for different immune related molecules on exosomes to induce T cell responses and tumor rejection in the B16 mouse melanoma model. Our data demonstrate ways to increase the feasibility of exosome-based therapeutic approaches in cancer.


Networking Lunch in the Exhibit Hall, Exhibits and Poster Viewing

Session Title: Exosomes/EVs, Circulating Cell-Free DNA (cfDNA) and Circulating Tumour Cells (CTCs) -- Circulating Biomarkers for Liquid Biopsy

Session Chairperson: Dr. John Ludlow, ZenBio


Sehyun ShinKeynote Presentation

PIBEX: Automated Extraction of Circulating Biomarkers from Plasma
Sehyun Shin, Professor & Director, Nano-Biofluignostic Engineering Research Center, Korea University and Anam/Guro Hospital of Korea University, Korea South


NanoView BiosciencesExosome Biomarkers: Multiparametric Characterization and Phenotypic Analysis
Clayton Deighan, Regional Sales Director, NanoView Biosciences

Unlocking the full diagnostic potential of exosomes is dependent, in part, on the ability to accurately analyze these small particles, to acquire proteomic data on the single-exosome level, and to do both for rare events. ExoView is uniquely positioned to measure and characterize single exosomes of the smallest sizes. ExoView provides quantification, sizing, and protein expression information, including surface and luminal proteins, all without the need for sample purification. A custom panel can be built to assay up to 6 biomarkers of interest.


Linking Pain-Regulated Exosomal-miRNAs to Target Expression in the Brain
Natasha Sosanya, Staff Scientist, USAISR, United States of America

Nerve injury in male rat’s results in differential expression (DE) of circulating exosomal-miRNAs (ex-miRNAs). The DE ex-miRNAs affect target expression and signaling pathways that contribute to the development of neuropathic pain.


Afternoon Coffee and Tea Break Plus Networking


Applying Phage-Displayed Random Peptide Libraries to Identify Exosomes as Biomarkers in Patients with Central Nervous System Disorders
Xiaoli Yu, Assistant Professor, University of Colorado Anschutz Medical Campus, United States of America

Like most other cells, brain tumor cells shed extracellular vesicles (EV) into their external environment and are considered potentially high-value biomarker reservoirs. We hypothesize that phage-displayed random peptide libraries can be applied to identify EV of CNS origin, and high affinity phage peptides can be used for rapid isolation and characterization of extracellular vesicles from blood of patients with brain tumors. Phage display has been successfully applied to many different areas of research, including immunology, cancer, drug discovery, epitope mapping, protein-protein interactions, and infectious diseases, targeting a broad cross section of protein families. Phage display technology is based on the construction of a polypeptide library fused to a bacteriophage coat protein. Phage are screened (“panned”) on surfaces coated with potential targets. We have applied two Phage-display peptide libraries (7mer &12 mer) and identified high affinity peptides specific to exosomes derived from brain tumor cell-lines and plasma of patients with other CNS disorders. The peptides shared high sequence homologies within the ones selected by the same panning targets as well as with others panned by exosomes from different sources. Our data suggest that phage peptide technologies can be used for enrichment, isolation, and characterization of EVs derived from different CNS cell types from blood of brain tumor patients and other CNS disorders.


Zen-Bio, Inc.Stem Cell Extracellular Vesicles as a Novel Therapy for Age-related Tendinopathy
John Ludlow, Executive Director, Regenerative Medicine, Zen-Bio, Inc.

Aging is the principal risk factor associated with tendinopathy and also contributes to a significant decrease in the ability to efficiently heal chronic tendon injury by altering the complex and highly coordinated processes required for normal resolution. In addition to increases in lifespan and a growing aging population that is remaining active, there is a concomitant rise in the prevalence of comorbidities and tendinopathy risk factors such as obesity and diabetes. These factors have led to a very high prevalence of tendon injuries in those over 60 years of age, reaching 50% of this population, resulting in decreased quality of life and an economic burden in the billions of dollars. The use of biological and stem cell therapies for tendon healing and repair have been widely investigated and have demonstrated some potential for soft tissue regeneration. Secreted extracellular vesicles (EVs), such as exosomes, are packed with potent pro-repair proteins and RNA cargos that are both cell type-specific, as well as differentially produced and secreted according to the cellular environment. We have demonstrated that mild heat shock improves the repair activity of stem cell-derived EVs in vitro and in vivo. By manipulating the cellular environment, we have found that we can alter stem cell EV production and secretion. Our results have validated an approach of manipulating the cellular environment in a closed-system bioreactor to modify the bioactive cargo of secreted EVs for improved tendon healing. This presentation will describe the further development of our pro-healing EV therapeutic approach to treat tendon injuries and tendinopathy specifically afflicting the elderly.


Extracellular Vesicles Spread Neurotoxicity in Amyotrophic Lateral Sclerosis
Davide Trotti, Professor, Scientific Director, Weinberg ALS Center, Vickie and Jack Farber Institute for Neuroscience, Thomas Jefferson University, United States of America


Particle Metrix, Inc.Phenotyping Extracellular Vesicles Using Tetraspanins and Fluorescence-NTA
Ross Jacobson, Technical Sales Leader, Particle Metrix, Inc.

Nanoparticle Tracking Analysis (NTA) measures size and concentration of nanoparticles in the size  range  from  10  nm  to  1  µm.  While  getting  a  particle  size  distribution,  the  user  typically cannot  discriminate  whether  the  particle  is  a  vesicle,  protein  aggregate,  cellular  trash  or  an inorganic  precipitate  from  scatter  data  alone.  The  fluorescent  detection  capability  of  NTA enables the user to gain biochemical information about the particle and increases the resolution and the discrimination power. NTA instruments traditionally are equipped with one laser wavelength in operation. This limits the detection of biomarkers to only a couple fluorescence dyes. To expand the number of excitation wavelengths, it was necessary to change of the laser  source,  a  time  and sample  consuming  step.  To  overcome  this  issue  Particle  Metrix developed a unique QUATT laser technology and now allows the rapid analysis of biomarker concentrations or ratios with four excitation lasers on the same sample.  Here we report the quick and easy EV Phenotyping via multi-wavelength f-NTA on the new Particle Metrix four laser system. Alignment-free switching   between excitation wavelengths and   measurement mode (scatter and fluorescence) allows quantification of biomarker ratios such as Tetraspanins (CD63, CD81, CD9 and CD41) on the same sample within minutes reducing measurement time and precious sample volumes.


Samir EL-AndaloussiKeynote Presentation

EV Engineering For Biomedical Applications
Samir EL-Andaloussi, Associate Professor, Karolinska Institutet, Sweden

The presentation will describe work-flows for scalable purification of bioengineered EVs, down-stream vesicular analytics and application of such EVs in therapeutic contexts.


Networking Reception with Beer, Wine and a Light Dinner in the Exhibit Hall


Close of Day 1 of the Conference

Tuesday, 18 February 2020


Morning Coffee, Tea and Pastries in the Exhibit Hall

Session Title: The Study of Various Classes of Circulating Biomarkers -- Challenges and Opportunities in the Field


Michael HellerKeynote Presentation

Abundance of Tumor Related Mutations in High Molecular Weight cf-DNA Isolated from PDAC Cancer Patient Plasma Samples
Michael Heller, Distinguished Scientist - Knight Cancer Institute at Oregon Health & Science University (OHSU), Center for Cancer Early Detection and Research (CEDAR); Professor, University of California-San Diego, United States of America

The availability of enough tumor related cell free (cf) DNA in patient blood samples could be a limitation for viable early cancer detection. Presently, next generation sequencing techniques focus on the detection of mutations in the smaller (<200bp) cancer related apoptotic cf-DNA fragments. We now have considerable evidence that cancer related mutations are also present in the higher molecular weight (>200bp) cf-DNA, which is often found at elevated levels in cancer patient blood samples. Use of a new electrokinetic chip device allows isolation and multi-omics analysis of the exosome protein biomarkers, as well as the high molecular weight (hmw) cf-DNA present in the “same” 50 µL blood, plasma or serum sample. The exosome protein biomarkers are first detected directly by on-chip immunofluorescent analysis. Subsequently, the hmw cf-DNA is eluted from the chip, whereupon digital PCR (dd-PCR), PCR and sequencing analysis is conducted to identify cancer-related point mutations. In a blinded pancreatic ductal adenocarcinoma cancer (PDAC) patient study, we were able to show correlation of elevated levels of glypican-1 and cf-DNA in PDAC positive samples, with the presence of KRAS mutations in hmw cf-DNA as determined by digital PCR and Sanger sequencing. These results provide support that a relatively large percentage of the hmw cf-DNA in the patient blood/plasma is coming from the tumors. Since low copy number of tumor related cf-DNA may be a limitation for early detection, the fact that the mutations exist in the hmw cf-DNA could be very important. This would of course have considerable implications for NGS, which preferentially tends to focus on mutation detection in the smaller cancer related apoptotic cf-DNA fragments. We now envision a future strategy for Multi-Omic analysis with on-chip exosome biomarker detection and PCR amplification, with option to do dd-PCR, Sanger sequencing and/or more encompassing NGS mutation and epigenetic analysis. The goal is to provide seamless sample-to-answer point-of-care liquid biopsy diagnostics, therapy monitoring and ultimately early cancer detection from a small sample of patient blood.


RealSeq BiosciencesHighly Sensitive Direct Quantification of cf-microRNAs for Biomarker Profiling by Next Generation Sequencing (NGS)
Sergio Barberan-Soler, CEO, RealSeq Biosciences

Cell-free circulating microRNAs (cf-miRNAs) are promising diagnostic and prognostic biomarkers for cancer and other diseases. cf-miRNAs are highly stable in biofluids and are therefore attractive as potential biomarkers via NGS-based detection compared to circulating DNA and protein markers. However, NGS detection of cf-miRNAs suffers from a high level of incorporation bias when sequencing libraries are prepared using the most widely used commercial kits. SomaGenics RealSeq library prep platform addresses this issue, with proven best-in-class accuracy of detection. The RealSeq platform includes a kit designed to profile miRNAs from biofluids (RealSeq-Biofluids) that typically detects four times as many cf-miRNAs as commercially available technologies. The latest kit in the RealSeq platform is RealSeq-T, the first targeted sequencing approach to detect panels of cf-miRNAs (up to 1,200) directly from plasma, with no organic extraction needed. Nearly 95% of sequenced reads from RealSeq-T libraries mapped to the miRNA panel. With these new capabilities, the RealSeq platform should advance the prospects of cf-miRNAs as clinical biomarkers.


Sequencing of cfDNA to Monitor Treatment Response in Cancer Patients
Taylor Jensen, Director, Research and Development, Sequenom (a LabCorp Company), United States of America


Deciphering the Vesicle Code: Making Sense of EV Heterogeneity
John Nolan, Professor, The Scintillon Institute, United States of America

Cells release extracellular vesicles (EVs) that can carry molecular cargo, including lipids, proteins, and nucleic acids, to nearby or distant cells and affect their function. Understanding the mechanisms of EV biogenesis, uptake and transport has the potential to lead to new biomarkers, diagnostics, and therapeutics. A limit to realizing this potential is the ability to measure individual EVs and their cargo quantitatively and reproducibly. Conventional bulk biochemical analyses, which report only the total amount of cargo in an EV preparation, cannot effectively assess compositional heterogeneity. Single EV analysis methods are required, but conventional tools are challenged to accurately measure small, dim particles. We have developed a vesicle flow cytometry (vFC) method that can detect and size individual EVs to 70 nm, and measure surface cargo to ~25 molecules/EV. vFC allows the identification and characterization of vesicle sub-types within heterogeneous populations in culture supernatants of biofluids. Single EV analysis using vFC reveals striking differences in the expression of canonical vesicle cargo such as tetraspanins, as well as other surface markers including integrins, tumor markers, and carbohydrates on EVs in biofluids and even EVs collected from cultured cell lines. vFC-based single EV immunophenotyping will enable us to understand the origins and functions of EVs in much the same way cell-based immunophenotyping has enabled a detailed understanding of the immune system, and lead to a new generation of EV-based biomarkers, diagnostics, and therapeutics.


Networking Coffee Break in the Exhibit Hall -- Visit Exhibitors and View Posters


Lydia SohnKeynote Presentation

Title to be Confirmed.
Lydia Sohn, Professor of Mechanical Engineering, University of California-Berkeley, United States of America


Networking Lunch in the Exhibit Hall


Capturing Heterogeneous Circulating Tumor Cells in Breast and Lung Cancer Using Nanotube-CTC-Chip
Balaji Panchapakesan, Professor, Department of Mechanical Engineering, Worcester Polytechnic Institute (WPI), United States of America

The ability to detect cancer and its heterogeneity non-invasively in the earliest of stages can make a significant clinical impact. Therefore, biopsy of blood and other bodily fluids also called “liquid biopsy” is an emerging concept for non-invasive detection and capture of circulating tumor cells. Circulating biomarkers is a rapidly growing field of research. Circulating biomarkers include, circulating tumor cells (CTCs), exosomes, circulating tumor DNA (ctDNA), cell-free DNA and RNA (mRNA, miRNA, non-coding RNA, and other RNAs), circulating proteins and metabolites. However, the clinical impacts of liquid biopsy are yet to be realized. We describe the nanotube-CTC-chip for capture and analysis of heterogeneous circulating tumor cells. Using a micro-array technology, well established in the clinical domain, and utilizing nanosurfaces, we demonstrate the capture and isolation of invasive CTCs and CTCs of multiple phenotypes in breast and lung cancer patients.


Mehmet TonerKeynote Presentation

Title to be Confirmed.
Mehmet Toner, Helen Andrus Benedict Professor of Biomedical Engineering, Massachusetts General Hospital (MGH), Harvard Medical School, and Harvard-MIT Division of Health Sciences and Technology, United States of America

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