08:00 | Conference Registration and Materials Pickup |
| Session Title: Latest Trends in Exosome Research |
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| Session Chair: Dr. Sasha Vlassov, Senior Staff Scientist, Life Technologies |
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09:00 | | Keynote Presentation Microvesicle (exosome) RNA as Biomarkers for Disease Johan Skog, Chief Scientific Officer, Exosome Diagnostics Inc, United States of America
Microvesicles (exosomes) are released from all cells and can be isolated from biofluids, including serum, plasma, urine, and CSF. These exosomes carry RNA (mRNA, microRNA, and other small RNAs) and protein from the donor cell that can be easily isolated even after long-term storage in freezers. The use of exosomal RNA has sparked a tremendous interest in the field of biomarkers, where access to the genetic status of disease in a non-invasive way is highly desirable. To develop a clinical test using microvesicle RNA, isolation of exosomes in a robust way that is easy, scalable, reproducible and allows for high-throughput processing in a clinical setting is critical. The RNA needs to be of high quality with markers that allow quality assessment of clinical samples. The use of exosomes is a great platform that enables longitudinal monitoring of a variety of mutations and transcript levels in tumors using qPCR, NGS and digital PCR. |
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10:00 | Coffee Break and Networking in the Exhibit Hall |
11:00 | Plasma Vesicles Content as a New Sample Type for Clinical Diagnosis and Prognosis of Myocardial Infarction and Stroke Dominique PV de Kleijn, Professor Experimental Vascular Surgery, Professor Netherlands Heart Institute, University Medical Center Utrecht, The Netherlands, Netherlands
Extracellular vesicles, including microvesicles, microparticles and exosomes are abundant in plasma. All cells secrete extracellular vesicles with release being very rapid after innate immunity activation of inflammatory cells. These vesicles are important in cell-to-cell communication in a variety of processes including coagulation, antigen presentation and tissue damage. Plasma extracellular vesicles can be easily isolated from frozen plasma or serum and contain protein, miRNA and RNA depending on source and stimulus. Isolating these vesicles from the blood and studying their content may provide rapid information on the pathological status of tissues. We showed that plasma vesicles content can be used for the diagnosis of Acute Coronary Syndrome in an emergency department cohort of 471 patient with chest pain. Next to this, we showed that vesicle content in a cohort of 1060 patients can be used for the prognosis of secondary cardiovascular events on top of existing risk factors and are expecting to have a CE certified kit available this year. We anticipate that plasma extracellular microvesicles in the near future will emerge as an important source for diagnostic and prognostic markers for cardiovascular disease and will help us in understanding the underlying biology.
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11:45 | Patients’ Blood miRNA Profiling as a New Tool for Prognosis in Multiple Myeloma Flavia Pichiorri, Assistant Professor, The Ohio State University, United States of America
Multiple Myeloma (MM) is an incurable cancer where prognosis is determined by the combination of the International Staging System (ISS) and fluorescent in-situ-hybridization (FISH) analysis of MM plasma cells (MM-PCs). However, the outcome predictions based on the current criteria are not precise and patients with similar characteristics have different outcome, thereby supporting the need for novel and more accurate prognosticators. microRNAs are short- non-coding-RNAs whose expression levels change in the transition from normal PCs to MM-PCs. miRNAs are master regulators of gene expression and have been found altered in all cancers. Their identification in body fluids suggests their possible use as novel biomarkers in MM patients. A comprehensive profile of circulating miRNAs for disease prognostication has not been reported. We profiled more than 654 different miRNAs using nCounter technology (nanoString, Seattle, USA) on samples from 54 newly diagnosed MM patients enrolled on the randomized phase 3 study of Velcade Melphalan Prednisone Thalidomide versus Velcade Melphalan Prednisone (NCT#01063179). Stem loop real-time PCR (qRT-PCR) was performed on an additional 234 MM patients enrolled in the same trial. We found that 3 miRNAs [miR-16 (P=0.0019), -19b (P=0.043) and miR-25(P=0.0012)] were associated with overall survival (OS) of myeloma patients. By incorporating these indices into the ISS score, we generated a novel miRNA-integrated score that is highly predictive of survival duration (P<0.00001) and substantially improved the currently used prognostic scores. These observation are perfectly aligned with our previously published data in which these miRNAs, are tightly regulated in MM cells and are shown to regulate their survival. We can then conclude that circulating microRNAs can be considered a new class of non-invasive prognosticators in MM.
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12:15 | Lunch and Networking in Exhibit Hall |
12:45 | Technology Spotlight: New High-throughput Biomarker Discovery Platform for Exosome Isolation and poly(A)+ mRNA Analysis Masato Mitsuhashi, Chief Scientific Officer, Hitachi Chemical Research Center, Inc.
We have developed a unique 96-well filterplate which captures exosomes and microvesicles (EMV) from various biological fluids, such as plasma, urine, saliva, cell culture media, etc. An additional adaptor will accommodate large volumes of culture media and urine (~10-30 mL). After centrifugation for only 5 min at 2,000 x g, EMV can be trapped on the membrane. For mRNA analysis, EMV is lysed on the filterplate, and the resultant lysates are transferred to a 96-well oligo(dT)-immobilized microplate for poly(A)+ mRNA isolation followed by cDNA synthesis and PCR. The results are comparable to those of standard ultracentrifugation method. In urine, glomeruli-, proximal tubules-, distal tubules-, and collecting ducts-specific mRNAs can be quantified, and are currently used for the analysis of kidney function in areas such as diabetic nephropathy and kidney transplantation. In plasma, various cell/tissue-specific mRNAs can be quantified, including hematopoietic cells in bone marrow, endothelial cells, neuronal cells in brain, skeletal muscle cells, fat cells, etc. The identification and quantification of tissue/cell specific mRNAs in EMV are applicable to various biomarker discovery projects. |
13:15 | Poster Viewing and Networking Session in Exhibit Hall |
| Session Title: LDTs, CLIA, and Opportunities in the Biomarker Interrogation Space |
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| Session Chair: Hannah Mamuszka, Vice President of Business Development, Exosome Diagnostics |
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14:15 | Companion Diagnostics Development at the Siemens Clinical Laboratory Sunil Pandit, Senior Manager, Siemens Healthcare Diagnostics, United States of America
Siemens Clinical Laboratory (SCL) is an accredited CLIA laboratory within Siemens Healthcare Diagnostics Inc., a global diagnostics company. SCL provides external access to laboratory developed tests (LDTs) and cutting-edge diagnostic technologies, pharmaceutical clinical trial testing services, and a unique menu of clinical tests. Utilizing LDT assays as prototypes for globally distributed In Vitro Diagnostic (IVD) products allows efficient development of new technologies and validated assays for companion diagnostics applications and clinical utility studies. In this presentation, the speaker will describe experience gained in developing companion diagnostics assays at Siemens. |
14:45 | Cancer Biomarker Discovery: The Case for Pharma OutSourcing from a Dx Perspective Premal Shah, Director, Genomic Health Inc, United States of America
Cancer therapeutic development is hard. Really hard. The successes have been few and far between. Many companies are looking to identify and incorporate biomarkers as part of their development plan. However, while companies often fixate on simple, “kittable” markers, the more powerful markers of the future—driven by leveraging NGS capabilities—will likely be complex. Identifying these markers requires niche expertise that can be provided only by companies with deep experience in this area. It is for this reason, combined with the costs that make outsourcing cancer biomarker discovery a necessity for today’s pharma companies. |
15:15 | Coffee Break and Networking in the Exhibit Hall |
15:45 | Extracorporeal Elimination of Circulating Exosomes as a Therapeutic Adjunct to Address Infectious Disease and Cancer James Joyce, Chairman & CEO, Aethlon Medical, Inc., United States of America
The Aethlon Hemopurifier® is a first-in-class device that targets the rapid elimination of infectious disease and cancer glycopathogens from circulation. In cancer, the Hemopurifier has been demonstrated to capture exosomes underlying lymphoma, melanoma, ovarian, and breast cancer. These microvesicular particles trigger apoptosis of immune cells and have been reported to facilitate tumor growth, metastasis, and the development of drug resistance.
In design, the Hemopurifier consists of the affinity lectin Galanthus nivalis agglutinin (GNA) immobilized in the outer-capillary space of plasma membrane technologies. The resulting mechanism provides selective target elimination as GNA binds high mannose signatures abundant on the surface of exosomes and viral glycoproteins. Studies of hepatitis c (HCV) infected patients receiving a three-treatment Hemopurifier protocol combined with interferon-based standard of care resulted in undetectable HCV in as little as seven days in hard-to-treat genotype-1 patients. The studies also documented the ability of the Hemopurifier to capture as many as 300 billion HCV copies during a single six- hour treatment. The FDA recently approved an IDE that allows for the initiation of US feasibility studies. |
16:15 | | Keynote Presentation Noninvasive Cancer Personal Genomics Charles Cantor, Chief Scientific Officer, Sequenom Inc, United States of America
In clinical situations where there is significant programmed cell death like cancer or pregnancy, DNA and RNA molecules appear in the circulation derived from cells that have died. In pregnancy, mature diagnostic tests are now available. In cancer, things are the situation is at a much more preliminary stage. To utilize DNA from the apoptotic cells it must differ in some way from the considerable background of host DNA. These differences can be copy numbers, sequence variations or epigenetic variations. In this talk I will contrast the situations in the two indications and chart possible future developments. |
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17:15 | Exosome Biogenesis: Role of Exosomal Cargo in the Production and Secretion of Exosomes Suraj Bhat, Professor, University Of California Los Angeles, United States of America
The molecular information content of exosomes is fundamental to their function. Exosomes, carry cell-type specific cargo targeted for extracellular destinations. While a constitutive synthesis of exosomes is conceivable as part of the cellular basis of tissue homeostasis we assume that the production of exosomes containing cell type-specific gene products may not be random but dictated by tissue physiology and cellular function; exosomes may be assembled and secreted in response to instructions received from the neighboring cells or from distant tissues. Thus an efficient way of coordinating the process of exosome production and secretion, which is temporally precise and functionally relevant, is to have the cell-type specific cargo a role in their biogenesis. We have used the human retinal pigment epithelium (RPE) as the functioning paradigm. RPE straddles the blood/retina barrier and physically and physiologically supports retinal photoreceptor function. Working with aB-crystallin (aB), a small heat shock and cell-type specific exosomal protein associated with a number of neuro-degenerations including Alzheimer’s, Parkinson’s disease, Multiple sclerosis and AMD (age-related macular degeneration), we demonstrate that this protein is essential for exosome biogenesis in the human RPE cells in culture. aB is found only in some cell types (for example most epithelial cells contain aB while most fibroblastic cell lines do not). Its presence in exosomes, therefore, allows us to address the question if the exosomal cargo has a role in exosome biogenesis? Understanding cell-type specific control of exosome biogenesis may allow us engineer exosomal content biologically. We will discuss our recent data obtained from these investigations. |
17:45 | Reception and Roundtable Discussions in Exhibition Hall |
19:15 | Close of Day One of Summit. |
19:15 | Exosome Night: Dinner: Premium Beers, California Wines, and Dinner for All Registered Exosome Track Delegates. Enjoy an Awesome Evening and Network with your Fellow Delegates. Sponsored by Exosome Diagnostics, Inc. |