Sunday, 26 October 201409:30 | Conference Registration and Coffee | |
Session Title: Introduction to the ISEV2014 Educational Event |
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Conference Chairman: Jan Lötvall, M.D., Ph.D. |
| | 10:00 | Welcome to the ISEV2014 Education Event by the President of ISEV
Jan Lötvall, Professor, University of Gothenburg; Founding President of ISEV; Editor-in-Chief, Journal of Extracellular Vesicles, Sweden
The President of ISEV will provide an introduction to the International Society for Extracellular Vesicles (ISEV) and will describe the goals and objectives of ISEV.
- Introduction to the ISEV Academic Society
- Topics and Areas to be Addressed during the ISEV2014 Educational Event
- How do EVs Synergize with Liquid Biopsies
| 10:15 | Introduction to ISEV2015 Washington DC
Kenneth Witwer, Associate Professor, Johns Hopkins University School of Medicine, United States of America
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Session Title: Isolation of Extracellular Vesicles (EVs) |
| | 10:30 | Isolation and Purification of EV and EV Subsets
Kenneth Witwer, Associate Professor, Johns Hopkins University School of Medicine, United States of America
Isolation and purification of extracellular vesicles and related particles, including viruses, has traditionally been performed by stepped centrifugation methods. Indeed, ultracentrifugation techniques remain standard in the field. However, the growing interest in EV as biomarkers and in therapeutics has been accompanied by a rapid proliferation of kit-based methods. Additionally, diverse combinations of approaches have been published in recent years. In this discussion, we will compare and contrast common methods for EV isolation and purification. The need to tailor the chosen approach to characteristics of the isolation medium and the ultimate research questions will be addressed. Finally, as an example, we will follow the crucial steps and variables in a representative workflow: from collection of a sample through EV purification and initial characterization. | 11:15 | Pitfalls and Prospects for Single EV Analysis using Flow Cytometry
John Nolan, CEO, Cellarcus Biosciences, Inc., United States of America
| 12:00 | Lunch, Networking and Poster Viewing | |
Session Title: Biomarkers in Extracellular Vesicles (EVs) |
| | 13:00 |  | Keynote Presentation The Diversity of Extracellular Vesicles and Their RNA Cargo
Jan Lötvall, Professor, University of Gothenburg; Founding President of ISEV; Editor-in-Chief, Journal of Extracellular Vesicles, Sweden
Most cells have the capacity to release different types of extracellular vesicles, which typically have an intact cell membrane which protects the cytoplasmic vesicular cargo. These extracellular vesicles include exosomes, microvesicles as well as apoptotic bodies, where the exosomes are the truly nano-sized vesicles, with a diameter range of 40-100 nm. In 2007, we were first to show that exosomes released from mast cells contain both microRNA and mRNA, which subsequently can be transferred from one cell to another, and mediate functionality. Importantly, the RNA cargo in exosomes are not directly related to the exosome-producing cell RNA, suggesting that specific mechanisms are involved in packaging RNA into exosomes. Subsequently, it has been shown that exosomes in all human body fluids investigated do contain RNA, including serum, plasma, saliva, semen and breast milk. Importantly, the RNA content in exosomes changes when the cells undergo for example oxidative stress. The capacity of exosomes to shuttle RNA between cells, has of course increased the complexity by which cells communicate with each other. Thus, this level of communication is significantly different from the classical communication pathways, including protein-receptor interaction as well as neural transmission. Importantly, different extracellular vesicles contain different RNA species, as well documented in different studies using different models. Therefore, cells may mediate multiple RNA-mediated signals by shuttling also other extracellular vesicles than exosomes to other cells. This presentation will primarily discuss the different RNA cargo in subsets of extracellular vesicles, including apoptotic bodies, microvesicles and exosomes, but also in relation to subgroups of vesicles within the exosomes population, and relate the studies to putatively clinically important effects of the vesicles. |
| 13:45 | EVs as Cancer Biomarkers
Dolores Di Vizio, Professor, Cedars Sinai Medical Center, United States of America
Our group and others have demonstrated that cancer cells release oncogenic cargo in extracellular vesicles (EVs). Transit of EVs through tissue spaces can alter the tumor microenvironment and elicit behavioral responses by cells exposed to them, such as cell motility, invasiveness and metastatic propensity. These discoveries point 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. Several approaches toward development of a “liquid biopsy” for cancer have been attempted. It has been known for years that both DNA and RNA can be detected in the circulation, and that circulating, cell-free (cf) DNA is more abundant in cancer patients than in controls, and can be an indicator of resistance to therapeutic regimens. Circulating DNA is also present in EVs derived from normal and tumor cells. The possibility that EVs preserve the stability of extracellular nucleic acid in the bloodstream is stimulating efforts to use EV fraction(s) in blood as a non-invasive source of personalized markers of disease aggressiveness, and as a means of following cancer progression and regression in real time. Our team recently reported that silencing of the gene encoding the cytoskeletal regulator Diaphanous related formin-3, DIAPH3, in tumor cells results in a transition to a rapid migratory phenotype characterized by dynamic membrane perturbations and increased metastatic potential. We also discovered that DIAPH3 silencing results in the export of large (1-10 µm diameter) bioactive EVs (large oncosomes) that originate from the shedding of bulky membrane protrusions from the plasma membrane. We have demonstrated that the abundance of large oncosomes in the circulation and in tissues correlate with advanced disease in mouse models and human subjects.
| 14:30 | Exosomic microRNAs as Orchestrators of the Biology of the Tumor Microenvironment
Muller Fabbri, Assistant Professor, University of Southern California, United States of America
MicroRNAs (miRNAs) are small non-coding RNAs involved in gene expression regulation. MiRNAs are dys-regulated in almost all types of human cancers, compared to the non tumoral adjacent tissue counterpart and affect all aspects of the biology of the malignancy. Recently, it has been shown that miRNAs are also secreted by cancer cells within exosomes and can be functionally transferred from one cell to another. This original observation has led to the identification of new, unexpected mechanisms of action for exosomic miRNAs. This presentation will focus on these new functions of exosomic miRNAs, showing how cancer cells affect the surrounding tumor microenvironment by secreting exosomic miRNAs. The discovery of this exosomic-miRNA-mediated cross-talk provides the rationale for new anti-cancer approaches. | 15:15 | Afternoon Coffee Break | 15:45 | Characterization of Glioma Exosomes
Bob Carter, Professor and Chief of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, United States of America
Glioma Associated Exosomes may be found in cerebrospinal fluid and blood. Recent studies have suggested that exosomes may be able to serve as biomarkers for glial neoplasms. This presentation will highlight current data, challenges and opportunities related to the use of exosomal biomarkers in clinical practice. | 16:30 | Considerations in Designing an ExRNA Biomarker Discovery Study
Louise Laurent, Associate Professor, University of California-San Diego, United States of America
Extracellular RNAs associated with exosomes and other extracellular particles are potential diagnostic and prognostic biomarkers. Here, we will discuss technical variables that should be considered when designing an ExRNA biomarker discovery study, including processes for biofluid collection, isolation and quantification of ExRNAs, and ExRNA analysis methods. | 17:15 | Functional Roles of Tumor-derived Extracellular Vesicles
Michael Freeman, Vice Chair of Research, Director, Cancer Biology and Therapeutics, Cedars-Sinai Medical Center, United States of America
Extracellular vesicles (EVs) include distinct subtypes of lipid-enclosed particles. They are involved in intercellular communication in many physiological and pathological conditions, including cancer. Tumor-derived EVs contain functional protein and nucleic acid cargo, which may induce tumor-promoting effects in nearby tumor cells. They can also modulate different types of cells in the tumor microenvironment and prepare other organ types for metastasis. EV populations are heterogeneous in composition, size and functional roles. This complexity is likely in part a reflection of the dynamic nature of gene expression and signal transduction that manifests in cancer and other pathologic states. | |
Session Title: Exosomes from Bacterial Species and EVs as Delivery Agents |
| | 18:00 | Bio-inspired Exosome-Mimetic Technology for Targeted Drug Delivery and Adjuvant-free Vaccine
Yong Song Gho, Professor, Pohang University of Science And Technology (POSTECH), Korea South
Communication between cells and environment is an essential process in living organisms. The secretion of extracellular vesicles is a universal cellular process occurring from simple organisms to complex multicellular organisms, including humans. Throughout evolution, both prokaryotic and eukaryotic cells have adapted to manipulate extracellular vesicles for intercellular communication via outer membrane vesicles in the case of Gram-negative bacteria and ectosomes or exosomes in eukaryotic cells. Recent progress in this area has revealed that extracellular vesicles play multiple roles in intercellular and interspecies communication, suggesting that extracellular vesicles are NanoCosmos, i.e., extracellular organelles that play diverse roles in intercellular communication. This presentation focuses on the our recent progress in novel exosome-mimetic technology for targeted delivery of chemotherapeutics and siRNA as well as for adjuvant-free, non-toxic vaccine delivery system against bacterial infection. | 18:45 | Extracellular Vesicle Round-Table Discussions with Beer
Jan Lötvall, Professor, University of Gothenburg; Founding President of ISEV; Editor-in-Chief, Journal of Extracellular Vesicles, Sweden
All Speakers and Conference Delegates are Invited to the Round-Table Discussions at the Conclusion of the Conference Presentations.
These Round-Table Discussions will discuss the most-pressing topics in the extracellular vesicles (EV) space as they exist today:
- Challenges in Isolation Methodologies
- Advantages and Disadvantages of Various Isolation/Purification Approaches for EVs from Various Sample Classes
- Interrogation of EV Biomarker Cargo [RNA, Proteins, DNA]
- Engineering of EVs
- EVs as Delivery Vehicles
- EVs in Regenerative Medicine & Cellular Therapy
| 19:30 | Close of Event |
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