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SELECTBIO Conferences Exosomes and Microvesicles:  Research, Biomarker Cargo and Therapeutic Potential

Exosomes and Microvesicles: Research, Biomarker Cargo and Therapeutic Potential Agenda



Other Track Agendas

Circulating DNA, Circulating RNA, Circulating Tumor Cells, Circulating Proteins | Exosomes and Microvesicles: Research, Biomarker Cargo and Therapeutic Potential | RNA-Seq, Digital PCR, and Liquid Biopsies: Capturing Value from Circulating Biomarkers | 

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Monday, 21 March 2016

08:00

Conference Registration, Materials Pick- and Continental Breakfast in the Exhibit Hall


Session Title: Opening Plenary Session


Plenary Session Chairman: John Palma, Ph.D.

09:00

John PalmaKeynote Presentation

Clinical Performance of Liquid Biopsy Based Detection of EGFR Mutations in NSCLC
John Palma, Chief Medical Officer, Roche Sequencing Solutions, United States of America

09:30

Chwee Teck LimKeynote Presentation

Microfluidics to Enable the Single Cell Analysis of Circulating Tumor Cells for Personalized Treatment
Chwee Teck Lim, NUS Society Chair Professor, Acting Director, Biomedical Institute for Global Health Research and Technology, National University of Singapore, Singapore

Cancer diagnosis and treatment are often hindered by the complexity of tumor heterogeneity.  Here, we developed a microfluidic biochip that is able to select any circulating tumor cell(s) of interest for downstream molecular analysis to probe their heterogeneity so as to obtain patient derived information for the personalized treatment of cancer patients.

10:00

Xandra BreakefieldKeynote Presentation

New Dimensions of Extracellular Vesicles – Cell Biopsies and Therapeutic Vehicles
Xandra Breakefield, Professor, Mass General Hospital (MGH)/Harvard Medical School, United States of America

The contributions of EVs to medicine continues to expand. These vesicles are virtual biopsies of living cells in the body that report on their dynamic physiology through an array of proteins, DNA and RNA.  This array is unique to EVs and accessible through biofluids, with profiles informative for different disease states and response to therapy. These vesicles can also be used as stealth vehicles to deliver bioactive agents, including drugs and genes. Their innate stability in vivo, robust loading capacity and ability to achieve efficient delivery to different tissues expands therapeutic modalities.

10:30

Coffee Break and Networking in the Exhibit Hall

11:00

Steve SoperKeynote Presentation

Integrated Microfluidic Systems for the Isolation of CTCs, cfDNA and Exosomes
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

11:30

Phil StephensKeynote Presentation

Will the Liquid Biopsy End FFPE Biopsy Testing to Guide Therapeutic Decisions for Patients with Cancer?
Phil Stephens, Chief Scientific Officer, Foundation Medicine, United States of America

There is tremendous excitement about the potential of “the liquid biopsy” to identify targeted therapeutic options for patients with cancer. However, for many tumor types,  significant and likely insurmountable challenges exist that currently exclude the liquid biopsy as an appropriate diagnostic specimen. This seminar will give a comprehensive overview of the complexities physician’s face in selecting the appropriate diagnostic assay to maximize treatment options for their patients.

12:00

Shana KelleyKeynote Presentation

New Approaches for CTC Subpopulation Profiling
Shana Kelley, Professor, University of Toronto, Canada

12:30

Networking Lunch, Discussions with Exhibitors and View Posters


Session Title: Biomarker Cargo of Extracellular Vesicles (EVs)

13:30

The Role of Extracellular Vesicles and their Content in Glioblastoma Microenvironment
Agnieszka Bronisz, Instructor in Neurosurgery, Brigham and Women's Hospital/Harvard Medical School, United States of America

Provide the overview on the role of extracellular vesicles (EVs) communication in glioblastoma microenvironment. Analysis of proteome and microRNA content of glioblastoma stem cells derived EVs. Present evidence for protein/microRNA functional transfer by EVs. Discuss the prospects of EV protein/microRNA-targeted strategy for brain tumor therapy.

14:00

Bob CarterKeynote Presentation

Exosomes and ExRNA Biomarkers for Glioma
Bob Carter, Professor and Chief of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, United States of America

14:30

iZON ScienceTechnology Spotlight:
Rapid isolation, Purification and Detailed Characterization of Extracellular Vesicles from Biological Fluids
Anoop Pal, Chief Scientist - Americas Region, iZON Science

Isolation and purification of extracellular vesicles (EVs) from complex biological fluids without changing their biophysical properties is critical. Moreover, accurate characterization of EV properties (size, size distribution, charge, mobility etc.) and quantification of EVs is paramount to link the impact of biophysical properties of EVs to biological outcomes.  This work presents a simple, repeatable, reliable and cost-effective approach that can be used for purification, isolation and detailed particle-by-particle characterization of EVs.

15:00

JSR Micro IncTechnology Spotlight:
New Tools for Exosome Isolation and Analysis: ExoCap Kit for Serum, Plasma & Cell Culture
Hiroaki Sagawa, Senior Manager, JSR Micro Inc

Research in circulating tumor cells, circulating cell-free DNA, extracellular RNA, and Extracellular Vesicles (EVs) have been gaining popularity as a promising non-invasive diagnostic approach. With this backdrop, JSR has focused on the development of EV capture methodologies, keeping in mind future potential uses for EVs in non-invasive diagnostics at the point of care. In this session, a description of a selective capture method of EVs as a potential platform for non-invasive diagnostics will be provided by introducing ExoCapTM, a magnetic beads based isolation and enrichment method of exosomes from various body fluids and cell culture supernatants. ExoCap consists of Magnosphere magnetic beads particles coupled with antibodies that recognize antigens on the exosome surface, a washing buffer, and a reagent that releases the captured exosomes for analysis. The antibodies against CD9, CD63, CD81, and EpCAM were specifically selected for this kit. ExoCap can easily separate exosomes, without ultracentrifuge or any special equipment. A sample amount of 0.1mL is sufficient. In addition, it is an animal free system which is superior to other methods for Mass Spec analysis. Moreover, this method enables non-destructive purification of exosomes. ExoCap can also be applied to beads based Flow Cytometry immediately after the capture process with 50 µL of samples. To confirm exosomal isolation from diverse body fluids (such as human serum, plasma) and cell culture media, exosomes were examined by western blot, RT-qPCR, particle size distribution measurement, Flow Cytometry, and scanning transmission electron microscopy (TEM). Exosomes isolated by ExoCap had a lipid bilayer membrane, showed a particle size distribution around 100nm, and expressed tetraspanin molecules.

15:30

Coffee Break and Networking in the Exhibit Hall

16:00

Stefan MommaKeynote Presentation

Extracellular Vesicle-Mediated Transfer of Functional RNA Between Tissues
Stefan Momma, Group Leader, University Hospital Frankfurt, Germany

Extracellular vesicles (EVs) emerge as important carriers for intercellular communication and they have been implicated in many biological processes. However, due to their small size and the difficulties of manipulating them, the data on EV biology has thus far been based mainly on in vitro or indirect in vivo evidence, or on experiments involving ex vivo manipulations. Furthermore, it is difficult to establish the exact cells of origin as well as individual cellular targets for in vivo experiments. Thus, the extent and pathogenic role of EV signalling in vivo, particularly with regard to the transfer of functional RNA, remains poorly understood. Using the Cre-Lox system we can overcome some of the major shortcomings in this field by establishing a method to trace the functional transfer of RNA by EVs in vivo.

16:30

Urinary Exosomes and Microvesicles as Carriers of Diagnostic Markers
Leileata Russo, Research Professor, Mass General Hospital (MGH)/Harvard Medical School, United States of America

Exosomes and microvesicles have been widely examined for their biological role in health and disease. This presentation will cover their potential use as a non-invasive ‘biopsy’ of the genitourinary tract and beyond. We have demonstrated that exosome and microvesicle RNA (collectively referred to as exoRNA) is extremely stable enabling reproducibility and enhancing sensitivity and specificity of analysis. An overview of the current use and reports of exosomes and microvesicles as a diagnostic platform will be given.

17:00

Enabling Urine Biomarker Discovery
Shannon Pendergrast, Chief Scientific Officer, Ymir Genomics, United States of America

The discovery of urinary extra-cellular biomarkers has been impeded by the lack of efficient methods for the isolation of extra-cellular vesicles (exosomes and microvesicles), protein, and nucleic acid.  Ultracentrifugation, considered the gold standard for vesicle isolation from many biofluids, is efficacious but laborious, and like most commercially available methods, is unable to consistently isolate enough quality material from small volumes of urine for protein or RNA-based biomarker discovery.  Ymir Genomics has invented a novel precipitation reagent that rapidly and efficiently isolates extra-cellular biomolecules and vesicles from urine samples yielding a quantity suitable for biomarker discovery.  Ymirite is more efficient than either ultracentrifugation or commercial precipitation methods for isolating extra-cellular nucleic acids and vesicles from a variety of urine samples as evaluated by immunoblot, Nanosight, Electron Microscopy, miRNA microarray, and qRT-PCR. Moreover, unlike all other methods, our reagent was able to isolate vesicles from very dilute samples. Ymir Genomic’s Precipitation Reagent offers a method that, in less than 30 minutes, isolates intact extra-cellular vesicles, protein and RNA suitable for biomarker discovery from urine samples.

17:30

QIAGENTechnology Spotlight:
From Sample to Insight: QIAGEN´s Approach Pioneering the Liquid Biopsy Revolution
Constanze Kindler, Senior Global Product Manager, QIAGEN

Liquid Biopsy has the potential to transform healthcare and biomedical research. The presentation will give an overview on QIAGEN’s approach to provide integrated solutions for isolation of circulating biomarkers and their analysis. The presentation will cover new and existing QIAGEN solutions ranging from sample stabilization, analyte purification, to analysis.

18:00

Networking Cocktail Reception for All Delegates, Speakers, Sponsors and Exhibitors

19:30

Close of Day 1 of the Conference. Extracellular Vesicles Dinner Short Course Begins.

Tuesday, 22 March 2016

07:30

Morning Coffee, Breakfast Pastries, and Networking


Session Title: Extracellular Vesicle (EV)-based Therapeutics

08:30

Evaluation of a Hybrid Exosome/Virus Vector System for Gene Transfer and Therapy in Preclinical Models of Human Disease
Casey Maguire, Assistant Professor of Neurology, Harvard Medical School, United States of America

Adeno-associated virus (AAV) vectors have shown remarkable efficiency in a number of preclinical models of disease in several organs including the eye, brain, muscle, heart, and liver. A major limitation to long term transduction, especially via the systemic route, is pre-existing immunity (humoral and cell-mediated) as well as the high vector load required to achieve sufficient levels of gene expression in the desired organ. Natural nanoparticles released from all cells, called exosomes, may have utility in creating better AAV vectors for human gene therapy. We have previously shown that harvesting AAV vectors from the media of 293T producer cells contains AAV vectors associated with exosomes (called exo-AAV). We have gone on to demonstrate that 293T-derived exo-AAV can evade neutralizing anti-AAV antibodies and greatly increase transduction in mice. Here we will report on our findings of the gene transfer capabilities of exo-AAV to several organs of interest in human disease including the brain, liver, muscle, eye, and cochlea.

09:00

Exosomes as Therapeutic RNAs Delivery Vesicles
Anastasia Khvorova, Professor, RNA Therapeutics Institute, University of Massachusetts Medical School, United States of America

Applications of RNA interference for neuroscience research have been limited by a lack of simple and efficient methods to deliver oligonucleotides to primary neurons in culture and to the brain. Here, we show that primary neurons rapidly internalize hydrophobic siRNAs (hsiRNAs) added directly to the culture medium without lipid formulation. We identify functional hsiRNAs targeting the mRNA of huntingtin, the mutation of which is responsible for Huntington’s disease, and show that direct uptake in neurons induces potent and specific silencing in vitro. Moreover, a single injection of unformulated hsiRNA into mouse brain silences Htt mRNA with minimal neuronal toxicity. Thus hsiRNAs embody a class of therapeutic oligonucleotides that enable simple and straightforward functional studies of genes involved in neuronal biology and neurodegenerative disorders in a native biological context.

09:30

Non-coding RNAs as Circulating Markers and Exosomal Therapeutic Targets in Neurological Disorders
Chaya Brodie, Professor, Henry Ford Hospital, United States of America

10:00

Peter  QuesenberryKeynote Presentation

Extracellular Vesicles as Biologic Modifiers
Peter Quesenberry, Professor of Medicine, The Warren Alpert Medical School of Brown University, United States of America

Extracellular vesicles are produced by essentially all tissues. Their nature is predicated by the nature of the originator cells, the treatment of the originator cells, the nature of the target cells and the type of initial vesicle separation. Vesicles have been shown to have deleterious or beneficial effects in different settings. If the vesicles derive from damaged or diseased tissue the effects are generally to make the situation worse, this is seen in cancer and pulmonary hypertension models. However, if the vesicles derive from normal tissues or mesenchymal stem cells healing effects are generally seen. Based on a number of studies it would appear that initial biologic effects of vesicles are due to transfer of mRNA and transcriptional regulators but long term effects are due to transcriptional modifications. Different studies have shown reversion of renal injury and radiation induced marrow injury with mesenchymal stem cell derived vesicles. The vesicles largely consist of exosomes and microvesicles which may be differently active in different models. The functional effects are stable on long term storage in 10% DMSO. Overall, administration of extracellular vesicles represent promising therapeutic approaches for a number of tissue injuries or diseases.

10:30

Coffee Break and Networking in the Exhibit Hall

11:00

Characterization of plasma Extracellular Vesicles with Human Age
Erez Eitan, Researcher, Laboratory of Neurosciences, National Institute on Aging Intramural Research Program, United States of America

Extracellular vesicles (EVs; also known as exosomes) are released from most cells into the extracellular environment and circulation. Thus, EVs play a potentially important role in intercellular communication between different cell types throughout the body.  Research has primarily focused on characterizing the proteins, lipids, and RNAs contained in EVs from various disease states, including cancer. As many of the pathological conditions where EVs have been implicated are age-related, it is likely that there changes in EVs with age in humans.  However, little is known about whether there are age-dependent changes in EV concentration (size and number), composition or function with human age.  We used a longitudinal cohort of 75 individuals from the Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) study to characterized EV concentration and surface proteins. We found a significant reduction in their concentration with age. Establishing a profile of EVs with human age will further aid in the development of diagnostics and therapies using EV-designed technologies for age-related diseases.

11:30

Gyongyi SzaboKeynote Presentation

Exosomes as Circulating Biomarkers for Liver Disease
Gyongyi Szabo, Professor & Vice Chair for Research, Department of Medicine, University of Massachusetts Medical School, United States of America

Exosomes are heterogeneous membranous vesicles originating from multivesicular bodies in most cell types in the body. Recently, they emerged as potential biomarkers or disease, effective conveyors of intercellular communication, and vehicles for delivery of nucleic acid based therapy. Moreover, both the biomarker potential and pathophysiological roles for exosomes are beginning to be recognized in various liver diseases including alcoholic liver disease, hepatocellular carcinoma, non-alcoholic fatty liver disease, hepatitis B  and hepatitis C viral infections.

12:00

Shannon StottKeynote Presentation

Microfluidic Liquid Biopsy: Isolation of Exosomes and Circulating Tumor Cells from Cancer Patients
Shannon Stott, Assistant Professor, Massachusetts General Hospital & Harvard Medical School, United States of America

Aggressive tumors will invade surrounding tissue due to rapidly dividing cancer cells that are nourished by an ample blood supply. As these cancer cells are multiplying, they release thousands of tiny particles into the blood stream, referred to as exosomes, which contain genetic information about the tumor. Whole cancer cells are also released from the tumor and these rare circulating tumor cells (CTCs) provide genetic and functional information about the patient’s cancer. Through a collaborative effort between bioengineers, biologists, and clinicians, our group at MGH has developed microfluidic devices to isolate both of these rare circulating biomarkers from whole blood. Data from these devices will be presented with a focus on our recent effort to characterize exosomes and CTCs from patients with glioblastoma. Through the microfluidic isolation of blood based biomarkers from glioblastoma patients, our goal is to obtain complementary data to the current standard of care radiologic measurements to help better guide treatment for this deadly cancer.

12:30

Networking Lunch, Discussions with Exhibitors and View Posters

13:00

Norgen Biotek CorporationTechnology Spotlight:
Lunch Technology Spotlight: Increasing the Sensitivity of Next-Generation Gene Expression Workflow for microRNAs from Bodily Fluids, Extracellular Vesicles and Small Sample Input - Sample Preservation, RNA Extraction and Sequencing Library Preparation
Bernard Lam, Senior Research Scientist, Norgen Biotek Corporation


Session Title: Emerging Themes and Approaches in the Extracellular Vesicles (EVs) Space

13:30

Heterogeneity of Cancer-Derived Extracellular Vesicles
Dolores Di Vizio, Professor, Cedars Sinai Medical Center, United States of America

Cancer cells release oncogenic cargo in bioactive extracellular vesicles (EVs) that can alter the tumor microenvironment and elicit behavioral responses in target cells. This discovery points to a role in tumor evolution for a conserved and finely regulated biological process that allows intercellular transfer of bioactive proteins, nucleic acids and lipids in the form of pre-assembled plasma membrane structures. In the last few years, it has emerged that every cell can release different types of EVs, with selective cargo and function. Studies on exosomes, for example, have demonstrated that diverse types of exosomes can be released by tumor cells. Several new methodologies are being interrogated to differentiate between exosomes with different cargo/functions. Different populations of exosomes as well as of larger EV types including the large oncosomes could provide clinically relevant information.

14:00

Stressing Out the Neighbors: Stressed Exosomes (“Sexosomes”?) Passage Stress Phenotypes to Recipient Cells (a Brief Proteomic Analysis)
Michael Graner, Associate Professor, Dept of Neurosurgery, University of Colorado Anschutz School of Medicine, United States of America

Cancer cells undergo a number of stresses, many of them self-inflicted, but often do not appear to suffer the consequences of those stresses. In some cases, the stress responses may actually prove beneficial to the tumor cells, providing them with potent resilience to their less-than-hospitable environments. One consistent tumor stress is the Unfolded Protein Response (UPR), an endoplasmic reticulum-based stress-management system with sensors, transducers, and effectors that result in a transcriptional and translational landscape rearrangement leading to resolution of the stress, or cellular apoptosis. However, tumors may incorporate the UPR into their stress portfolio to survive or even thrive amidst their environmental insults. We propose that exosomes from stressed cells (stressed exosomes, or “sexosomes”) are able to induce stress response phenotypes in recipient, unstressed cells, thus enabling stress responses without having to experience the actual stress. Our analysis in this report goes to the molecular level, monitoring proteome changes in glioma cells when those cells are exposed to exosomes released from UPR-stressed cells. We find high overlap in the proteomes of stressed cells and unstressed cells that receive “sexosomes”, suggesting that tumors may unify their overall stress responses despite their inherent heterogeneity. The implications for general tumor biology, and in particular, therapeutic resistance, are highlighted.

14:30

Microfluidic Phenotyping of Circulating Exosomes for Cancer Diagnosis
Yong Zeng, Assistant Professor, University of Kansas, United States of America

15:00

EGFR Signaling Regulates Extracellular Vesicle microRNA Contents
Alain Charest, Associate Professor, Dept of Neurosurgery, Tufts University Medical Center, United States of America

The focus of the research here is on leveraging clinically relevant genetically engineered mouse models of primary malignant brain cancer to study central aspects of gliomagenesis and molecular responses to therapeutic interventions. Recent work from the laboratory involves studies on the control exerted by EGFR signaling on extracellular RNA composition in Glioblastoma Multiforme.

15:30

Smartphone-based Optofluidic Exosome Diagnostic for Concussion Recovery
Jina Ko, Researcher, University of Pennsylvania, United States of America

We developed a smartphone-based optofluidic platform to measure brain-derived exosomes. Sample-to-answer on our chip is <1 hour and the key innovation is an optofluidic device that can detect enzyme amplified exosome biomarkers, and is read out using a smartphone camera. Using this approach, we profiled GluR2+ exosomes to detect and monitor concussion.

15:45

Short Coffee Break

16:00

Microfluidics for Exosome Analyses
Hakho Lee, Associate Professor, Director of the Biomedical Engineering Program, Massachusetts General Hospital / Harvard Medical School, United States of America

16:30

Nanoplasmonic Exosome (nPLEX) Analysis
Hyungsoon Im, Assistant Professor, Center for Systems Biology, Mass General Hospital (MGH)/Harvard Medical School, United States of America

This presentation will review an exosome analysis technology named nPLEX (nano-plasmonic exosome). The sensor is based on transmission surface plasmon resonance (SPR) through periodic nanohole arrays. Target-specific exosome binding to the array causes SPR signal changes, which enables sensitive and fast detection of exosomes. We applied the first generation nPLEX system to detect exosomes collected from ovarian cancer patients.

17:00

Potential Role of Exosomes in the Development of Malignant Mesothelioma
Phillip Munson, Researcher, University of Vermont, United States of America

Preliminary insights on the characterization of exosomes, and their cargo, from normal and asbestos exposed epithelial cells, as well as studies on the effects, and in vivo localization, of these exosomes on possible target mesothelial cells and tissues.

17:30

Mechanism and Application of ARRDC1-mediated Microvesicles (ARMMs)
Quan Lu, Associate Professor, Harvard School of Public Health, United States of America

I will discuss the molecular mechanism and physiological functions of ARMMs. I will also present data on how ARMMs may be used for therapeutic delivery of bioactive large molecules.

18:00

Close of Day 2 of the Conference


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