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SELECTBIO Conferences Extracellular Vesicles 2022: Biology, Disease & Medicine

Extracellular Vesicles 2022: Biology, Disease & Medicine Agenda

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Extracellular Vesicles 2022: Biology, Disease & Medicine | 3D-Models for Drug Testing: Organoids & Tissue Chips 2022 | 

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Tuesday, 13 September 2022


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


Lucia LanguinoConference Chair

Welcome and Introduction by Conference Co-Chairperson and an Overview of the Conference
Lucia Languino, Professor of Cancer Biology, Thomas Jefferson University, United States of America


Michael GranerConference Chair

Welcome and Introduction by Conference Co-Chairperson and an Overview of the Conference
Michael Graner, Professor, Dept of Neurosurgery, University of Colorado Anschutz School of Medicine, United States of America

Session Title: Conference Opening Session -- EVs in Biology and Medicine: Impact on Disease Classes


exRNA Analysis In Biofluids to Differentiate Diseases and Injuries of the Nervous System
Kendall Van Keuren-Jensen, Professor and Deputy Director, Translational Genomics Research Institute, United States of America

Our laboratory characterizes the exRNA cell-free and EV cargo in biofluids for the information they can provide about changes associated with disease, injury processes and progression.


Plasma Extracellular Vesicles For Cardiovascular 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. We use plasma extracellular vesicle (EV) protein content of vesicles from plasma sub-fractions on plasma of stroke and peripheral artery disease(PAD) patients, patients after carotid atherectomy (CEA) and patients suspected for chronic coronary syndrome (CCS). Using 25 ul of plasma, we developed an automated 96-well based protocol using sequential precipitation. Using samples of the AtheroExpress, the largest ongoing CEA biobank we try to early detect the risk of a second Major Adverse Cardiovascular Event (MACE: myocardial infarction, stroke or cardiovascular death) in PAD and CEA patients. Identification of such high risk patients is very important for possible (expensive) add-on pharmaceutical therapy or the decision to operate or not. Sequential precipitation for EV isolation is also used for the diagnosis of CCS.


Using Neuronal Enriched Extracellular Vesicles To Determine Brain Health
Lynn Pulliam, Professor of Laboratory Medicine and Medicine, University of California San Francisco, United States of America

Circulating plasma EVs can be captured by a neuronal surface protein (L1CAM) and the cargo interrogated to determine the health of neurons in real time. We have used protein arrays to look at neuronal EV cargo in people with HIV or post-COVID cognitive impairment.


Mid-Morning Coffee, Tea and Networking in the Exhibit Hall

Session Title: Technologies and Tools for Studying EVs

Session Chairperson: Professor Michael Graner, University of Colorado


Next generation nanoFACS with High Resolution Imaging and Custom 50um Nozzle
Terry Morgan, Professor, Oregon Health and Science University, United States of America

Nanoscale flow cytometry is an emerging technology designed to image, count, and isolate cell- and size-specific extracellular vesicles (EVs). More efficient approaches to nanoscale flow sorting (nanoFACS) are required for sufficient EV yield per volume for validation studies of sorting specificity and more precise characterization EV protein and nucleic acid contents.


Digital Flow Cytometry For Single-EV Analysis
Daniel Chiu, A. Bruce Montgomery Professor of Chemistry, University of Washington, United States of America

We have developed a multi-parametric high-throughput flow-based method for the analysis of individual extracelluar vesicles (EVs), which are highly heterogeneous and comprise a diverse set of surface protein markers as well as intra-vesicular cargoes. Yet, current approaches to the study of EVs lack the necessary sensitivity and precision to fully characterize and understand the make-up and the distribution of various EV subpopulations that may be present. Digital flow cytometry provides single-fluorophore sensitivity to enable phenotyping single EVs with unprecedented precision and sensitivity.


Networking Lunch in the Exhibit Hall with Exhibitors and Conference Sponsors


SpectradyneTrends in EV Characterization Technologies
Jean-Luc Fraikin, CEO, Spectradyne

Methods for characterizing EVs are evolving, and new technologies are being developed that deliver size, concentration, and fluorescent phenotyping, each to a varying degree of success.  Learn about how some of these new and cutting-edge technologies work, their strengths and weaknesses, and where we at Spectradyne see the technology landscape moving next.


IZON SciencePaving the Way to Automation and Standardization for Scalable Isolation of EVs
Jared Lynch, Director of Business Development, IZON Science

As the potential for EV-based diagnostics and therapeutics continues to grow, so too does the need for scalable isolation and precise characterization. In this presentation, I will present how technologies and services offered by Izon Science fill this gap. I will introduce Izon’s newly released Gen 2 qEV SEC isolation product line as well as our recent work on scaling up EV isolation and purification using larger sized qEV columns with automated flow systems. Finally, you will be taken through an overview of Izon platforms and offerings, to gain a clear understanding of how Izon technologies can fit into a larger workflow.


Particle Metrix GmbH and CEO, Particle Metrix Inc., USABiomarker Co-localization Measurements (C-NTA) Using the New Particle Metrix ZetaView x30 Family
Sven Rudolf Kreutel, Chief Executive Officer, Particle Metrix GmbH and CEO, Particle Metrix Inc., USA

During the last decades, Nanoparticle Tracking Analysis (NTA) has emerged as a vital and fast characterization technology for biological nanoparticles like Extracellular Vesicles, Exosomes and Viruses. While classic NTA scatter operation feeds back particle size and total concentration, the fluorescence detection capability enables the user to gain specific biochemical information. Determination of biomarker co-localization however, is a challenging task for any analytical instrument. A new laser generation ensuring perfect overlap of the illumination volumes of individual channels paired with very fast switching times between fluorescence channels lay the foundation of co-localization nanoparticle tracking analysis (C-NTA) introduced recently by Particle Metrix. For the first time, we report results of co-localization measurements of reference material as well as real biological samples based on NTA technology on the new ZetaView® PMX-230 TWIN.


NanoFCM Co., LtdNanoAnalyzer: Combining Flow Cytometry and Particle Analysis to Meet the Challenges of EV Analysis
Dimitri Aubert, Managing Director, NanoFCM Co., Ltd


Mid-Afternoon Coffee and Tea Break and Networking in the Exhibit Hall

Session Title: Conference Joint-Plenary Session


Nancy AllbrittonPlenary Presentation

Miniaturized Colon-on-Chip for Probiotic and Drug Screening
Nancy Allbritton, Frank and Julie Jungers Dean of the College of Engineering and Professor of Bioengineering, University of Washington in Seattle, United States of America

Organ-on-chips are miniaturized devices that arrange living cells to simulate functional subunits of tissues and organs. These microdevices provide exquisite control of tissue microenvironment for the investigation of organ-level physiology and disease. Planar models enable high throughput screening with primary human intestinal epithelial cells- both stem/proliferative cells and differentiated zones. Compound screening for stimulation or inhibition of hormone secretion by enteroendocrine cells is of high value for therapeutic development given the role that hormones and neurotransmitters such as glucagon-like peptide 1 and serotonin play in regulating human feeding behavior and metabolism. These simple models can also be adapted to produce a thick, functional mucus layer with or without an oxygen gradient to create an anaerobic luminal surface for culture of colonic flora. Such devices are high of value in evaluating the impact of probiotics- a growing therapeutic area for the treatment of human disease. These planar systems can be modified to produce “flat crypts” with a stem/proliferative zone and differentiated cell region for the study of stem cell proliferation, lineage allocation and migration. Fully 3D polarized epithelium possess an array of crypt-like structures replicating the intestinal architecture. Imposition of chemical gradients across the crypt long axis yields a polarized epithelium with a cell migration from a stem-cell niche into a differentiated cell zone. This in vitro human colon crypt array replicates the architecture, luminal accessibility, tissue polarity, mucus layer, cell types and cellular responses of in vivo intestinal crypts. These bioanalytical systems provide both high throughput as well as low throughput/content rich platforms for assay of microbiome-behavior, drug-delivery, and other assays with a living human intestinal epithelium.


Daniel ChiuPlenary Presentation

New Fluorescent Reagents to Enable Highly Multiplexed Single-Cell Measurements and Biological Assays
Daniel Chiu, A. Bruce Montgomery Professor of Chemistry, University of Washington, United States of America

Fluorescence based techniques have become an indispensible tool kit in both basic cellular studies and in vitro diagnostics. However, the intrinsic limitations of conventional dyes, such as short Stoke’s shift and low absorptivity, have posed difficulties for advancing highly multiplexed assays. We have developed a new class of fluorescent probes called Pdots, and this talk will highlight their development to enable high multiplex single-cell analysis and biological assays.


Danilo TaglePlenary Presentation

Developing Technologies for Single Vesicle Isolation and for Regenerative Medicine
Danilo Tagle, Director, Office of Special Initiatives, National Center for Advancing Translational Sciences at the NIH (NCATS), United States of America

Extracellular vesicles (EVs) are lipid membranous vesicles released from almost all cell types, and they provide a tremendous opportunity as sources of novel biomarkers from liquid biopsies, as well as agents for tissue repair and wound healing in regenerative medicine. EVs carry complex molecular cargoes, such as proteins, RNAs [e.g., mRNA and noncoding RNAs (microRNA, transfer RNA, circular RNA and long noncoding RNA)], and DNA fragments; these cargoes are delivered to recipient cells and serve as a cell-to-cell communication system. The molecular contents of EVs largely reflect the cell of origin and thus show cell-type specificity. Exosomes are endogenous nanoparticles that constitute a fraction of extracellular vesicles that are secreted by all cell types into the extracellular environment, and play an important role in intercellular signallng.  Exosomes are being utilized in a variety of biomedical applications, including targeted drug delivery, gene therapy, diagnosis, and tissue regeneration. Despite significant efforts made in this relatively new field of research, progress has been held back by challenges such as inefficient separation methods, difficulties in characterization, and lack of specific biomarkers. This presentation will elaborate on the NIH support for exosome isolation, as well as its potential use in regenerative medicine.


Luminex Technology Spotlight Presentation on EV Technologies


Networking Reception with Beer and Wine in the Exhibit Hall: Network with Colleagues and Engage with Exhibitors and Conference Sponsors


An Evening with Luminex -- Visit their Labs in Downtown Seattle, View Product Demonstrations, Enjoy Food & Drinks and Engage

Wednesday, 14 September 2022


Morning Coffee and Pastries in the Exhibit Hall

Session Title: EVs in Biology, Disease and Medicine

Session Chairperson: Professor Lucia Languino, Thomas Jefferson University


ILIAS Biologics Inc.Exosome Engineering for Active Targeting and Intracellular Delivery of Therapeutic Proteins: Realization from Concept
Chulhee Choi, CEO, ILIAS Biologics Inc.

As extracellular vesicles that play an active role in intercellular communication by transferring cellular materials to recipient cells, exosome offer great potential as a natural therapeutic drug delivery vehicle. Currently, both academia and industry try to develop exosome platform-based therapeutics for disease management, some of which are already in clinical trials. An optogenetically engineered exosome system (EXPLOR®) that we previously developed was implemented for loading therapeutic cargo into exosomes which can deliver therapeutic cargos into target cells in free form. We are studying the clinical potential of ‘Exo-target®’, therapeutic exosomes with EXPLOR technology, in multiple disease areas including inflammatory diseases. NF-kB has been well accepted as master regulator for inflammation. The introduction of IkB into target cells can inhibit inflammatory responses by restraining nuclear translocation of NF-kB. Therefore, we have developed Exo-target loaded with dominant active form of IkB (Exo-srIkB). We found that Exo-srIkB treatment attenuates both local and systemic inflammation and animal mortality associated inflammatory disease including sepsis, ischemic reperfusion induced acute kidney injury and preterm birth in our preclinical studies. Especially, maternally injected Exo-srIkB could cross-over placenta barrier to deliver therapeutic cargos to fetal side, which resulted in prolonged pregnancy by more than 24 hours and additional advantages. Altogether, these results suggest therapeutic value of Exo-target for various disease by delivering API intracellularly to the target cells.


Cancer-derived Extracellular Vesicles as Early Disease Biomarkers
Andreas Möller, Associate Professor, Group Leader and Head, Tumour Microenvironment Laboratory, QIMR Berghofer Medical Research Institute, Australia

Late diagnosis of cancer is a key cause of the high cancer-associated mortality. However, a most cancer progression occurs ‘silently’, associated with no or diffuse symptoms. A non-invasive test, for example using blood, to determine that an individual has an early stage cancer would provide curative therapeutic options and presents an important unmet clinical needs. In this presentation, I will summarize the research approaches in the area of cancer-derived EVs undertaken by my group. I will discuss novel data on how liquid biopsies based on small amounts of blood can determine the presence of early stage cancer and how this can be potentially applied clinically. In detail, we derived a cancer-specific multi-protein EV marker capable of detecting the presence of 8 cancer types in individuals, including those patients with stage I. Confirming the platform independence of the biomarker, we transferred the test to a SERS-based microfluidics platform, and demonstrate the capacity to distinguish patients with benign and malignant conditions. Overall, our biomarker is capable of detecting early stage cancer in the blood of individuals and has various potential applications in reducing cancer-associated mortality.


Use of Circulating Extracellular Vesicle Protein Biomarkers to Detect Disease Earlier
Jean Lewis, Associate Director, Research, Biological Dynamics, United States of America

Circulating extracellular vesicles (EVs) contain a wealth of biomarkers and are key to numerous emerging liquid biopsy approaches; information from these EV biomarkers can be harnessed for diagnosis of many different diseases.  For example, EVs released from tumors may carry biomarkers signaling the presence and type of cancer; EVs bearing markers of neurological damage can be found in circulating blood, and in the case of infectious disease, pathogen-related markers can be released on EVs from infected blood cells.  While there are multiple innovative, emerging technologies designed to measure EV biomarker levels, many rely on multi-step procedures for prior isolation of the EVs from plasma or serum or are very low throughput.  To advance EV biomarker analysis towards true point-of-care diagnostics, we have developed a lab-on-a-chip nanoparticle isolation platform (Verita™), designed to both isolate and immobilize extracellular vesicles directly from unprocessed blood fractions onto an AC Electrokinetics (ACE) microelectrode array.  The EV-associated biomarkers can be either eluted for further analysis or quantified directly on-chip, where the entire process takes less than an hour, showing the potential for point-of-care applications.  The Verita™ platform was used to screen clinical serum or plasma samples and demonstrated feasibility for detecting various disease types.  For example, performance of an assay using ACE-purified plasma EVs from stages I-II pancreatic cancer showed a sensitivity of 83% at 99% specificity.  EVs quantified directly on-chip can be probed with biomarkers related to early cancer or Alzheimer’s disease, and can be used to detect TB infection with an AUC of 1.00.


Mid-Morning Coffee, Tea and Networking in the Exhibit Hall


Extracellular Vesicles in Cancer Progression: Pro- or Anti- Tumorigenic?
Lucia Languino, Professor of Cancer Biology, Thomas Jefferson University, United States of America


Convective Transport and Binding of Extracellular Vesicles Establish Biologically Relevant Spatial Gradients
Steven C. George, Professor and Chair, Department of Biomedical Engineering, University of California-Davis, United States of America

Extracellular vesicles (EVs) are small (50-150 nm diameter) composite particles secreted by cells and comprised of a lipid-based membrane surrounding an aqueous core.  The membrane and core can each incorporate a wide range of molecules (e.g., proteins, nucleic acids) that can impact cellular function; thus, EVs can impact in vivo biology, but have also generated significant excitement for their potential theranostic (therapeutic and diagnostic) applications in cancer. How EVs are transported (convection, diffusion, and binding) within the extracellular space is poorly understood.  We hypothesized that EVs are transported predominantly by convection through the interstitium and could establish a spatial gradient via binding to laminin through integrins alph3beta1 and alpha6beta1 expressed on their surface. Our early experimental studies demonstrate EV binding to a laminin-rich extracellular matrix (ECM) increases as the malignant potential of the cellular source increases (MCF10A, MCF10DCIS, MCF10CA1). Binding of the EV to the ECM generates an observable spatial gradient, which impacts the migration of M2-like differentiated macrophages.  The magnitude of the gradient is partially abrogated by blocking antibodies to alpha3beta1 and alpha6beta1 integrin subunits. Examination of EV interstitial transport will enhance our understanding of the dynamic tumor microenvironment and could present new targets for early-stage disease.


Elisa WoodhouseKeynote Presentation

NCI-Supported Extracellular Vesicle Research: Perspectives and Opportunities
Elisa Woodhouse, Program Director, Tumor Biology and Microenvironment Branch, NIH/NCI, United States of America


Networking Lunch, Meet Exhibitors and Engage with Colleagues


Anti-Metastatic Activity of PRR851, A Specific Inhibitor of the Nuclear Transport of Extracellular Vesicles
Aurelio Lorico, Professor of Pathology, Touro University Nevada School of Medicine, United States of America

The metastatic process is responsible for the vast majority of cancer deaths, yet most therapeutic efforts have focused on targeting and interrupting tumor growth rather than impairing metastasis. Identifying and testing new pathways to disrupt the process of metastasis is the key to developing new therapeutic strategies that improve cancer survival. We have identified and described a novel cellular pathway that allows extracellular vesicles (EVs) derived from cancer cells to transfer oncogenic cargoes into the nuclear compartment of mesenchymal stromal cells and transform their gene expression. This pathway relies on the translocation of Rab7+ late endosomes, which contain endocytosed EVs, into the nucleoplasmic reticulum (NR) of recipient stromal cells. The interaction between the two organelles; late endosomes and the nucleus, is mediated by a novel tripartite protein complex (named VOR) formed by VAMP-associated protein A (VAP-A) in the outer nuclear membrane, cytoplasmic oxysterol-binding protein -related protein-3 (ORP3) and endosomal Rab7 GTPase. Silencing VAP-A or ORP3 in recipient cells prevents late endosome entry into the NR, thereby blocking EV-mediated intercellular communication. We have designed and synthesized new chemical entities (NCEs) that inhibit VOR complex formation and nuclear transfer of EV cargoes. We have identified a lead NCE (PRR851), that reduced the incidence of metastasis in a syngeneic mouse model of malignant melanoma without apparent off-target toxicity. We will discuss the structure-activity relationship of PRR851 and its analogs and new data in experimental metastasis systems.


Glioblastoma Extracellular Vesicle Specific Peptides Inhibit EV-induced Neuronal Cytotoxicity
Xiaoli Yu, Assistant Professor, Department of Neurosurgery, University of Colorado Anschutz Medical Campus, United States of America

Glioblastoma (GBM) is the most aggressive and lethal form of brain tumors. Tumor cells release extracellular vesicles (EVs) which have been shown to play a critical role in cellular communication in the tumor micro-environment. We discovered that GBM plasma EVs were smaller in size, had no relationship between size and concentration, but showed highly significant correlation between EV concentration and plasma protein concentration. Importantly, GBM EVs purified from both plasma and tumor cell line produced IgG-mediated, complement dependent cytotoxicity in neurons. We identified high affinity phage peptides for GBM EVs by screening phage-displayed random peptide libraries. Significantly, we showed that GBM EV peptides inhibit EV-induced complement-dependent cytotoxicity in neurons in dose- and time-dependent manner. Phage peptide technologies can be used for isolation and characterization of EVs derived from brain tumors. EV specific peptides can be explored for  biomarkers and mechanisms  of brain tumor EV-induced neuronal death.


Lung Transplant Immunology Through the Exosome Lens
Billie Hwang, Transplant Immunologist, University of Washington, United States of America

Lung transplantation improves survival and quality-of-life for patients with end-stage lung disease. Clinical risk factors post-transplant including primary graft dysfunction (PGD) and acute and chronic lung allograft dysfunction (ALAD and CLAD) negatively impact the long-term benefits of transplantation and continues to be an area of research that has not been fully understood. Exosome immunology is a novel area of research that directly implicates extracellular vesicles (30-150nm) in playing a major role in a wide variety of diseases and immune responses. Despite the explosion of exosome studies in the last 5 years, their role in solid organ transplantation has been limited. In our studies, we aim to understand the role of exosomes in the donor:host immune responses and as prognostic biomarkers in clinical outcomes post-transplant including the development of acute and chronic rejection. In addition, we aim to develop a potentially novel paradigm that redefines lung transplant immunology through an exosome lens.


Mid-Afternoon Coffee and Tea Break and Networking in the Exhibit Hall


Differential Effects of APOE Genotype on Cerebrospinal Fluid Extracellular Vesicle miRNAs in Females vs. Males with Alzheimer's Disease
Julie Saugstad, Professor, Department of Anesthesiology & Perioperative Medicine, Oregon Health & Science University, United States of America

Alzheimer’s disease (AD) is the most common form of dementia and the sixth-leading cause of death in the United States. We identified and validated miRNA biomarkers for Alzheimer's disease (AD) in cerebrospinal fluid (CSF) from living donors, then showed that five of the AD miRNAs trend down in expression from controls to mild cognitive impairment to AD. Our recent studies revealed that two biological risk factors for AD, sex and APOE e4 genotype, differentially effect CSF EV miRNA levels: APOE e4 within each sex altered distinct miRNAs, and predicted gene targets and pathways of the altered miRNAs are known to contribute to neurodegeneration in AD. We are currently establishing the physical features and molecular cargo of AD CSF EVs within the context of APOE genotype and sex, with the goal to identify novel gene targets of CSF EV miRNAs that are relevant to AD pathophysiology.

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