Exosomes and Liquid Biopsies Asia 2017 Agenda

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

Morning Coffee and Tea Break: Network with Your Peers and Poster Viewing

Cell-Free DNA Analysis of NSCLC Patients: A Paradigm for Clinical Use of Liquid Biopsies
Partha Das, Global Medical Affairs Lead, Genomics and Oncology, Roche Molecular Systems, United States of America

Networking Lunch and Meet Exhibitors and View Posters

From Seeing to Believing: Visualization and Tracking of Extracellular Vesicles
Charles Lai, Assistant Research Fellow, Institute of Atomic and Molecular Sciences, Academia Sinica, Taiwan

Extracellular vesicles (EVs), including exosomes and microvesicles, are nanosized membrane- bound vesicles released in abundance by cancer cells. These EVs are capable of delivering a select subset of proteins and RNAs designed to promote tumor growth and metastases. However, the exact time, location (i.e. spatiotemporal properties) and cargo delivery capacity of EVs remained unknown due to a lack of available methods to detect them. Here we engineered bioluminescent and fluorescent reporter systems which allow EV visualization in real-time both in vitro and in vivo. The multimodal bioluminescent reporter labels EVs with a membrane-bound variant of Gaussia luciferase fused to a biotin acceptor peptide, thereby enabling whole-animal multimodal imaging of nanoscale EVs, as well as pharmacokinetics analysis of EVs. We next developed the bifunctional fluorescent EV/EV-RNA reporters to study multiple EV populations with subcellular image resolution. By multiplexing the fluorescent and bioluminescent EV reporters, we further elucidated the rapid dynamics of tumor EV uptake and translation of EV-delivered cargo mRNAs in cancer cells. These novel molecular imaging platforms revealed that EV mediates a dynamic and multidirectional communication between cells with delivery of functional mRNA, thereby manipulating its recipient cells at neighboring and distal sites.

Panel Discussion: The Various Circulating Biomarker Classes and Their Utility for Liquid Biopsy Development: Circulating Cell-Free DNA (cfDNA), Circulating Tumor Cells (CTCs), and Circulating Extracellular Vesicles (EVs, Exosomes)

Close of Day 1 of the Conference Formal Presentations and Networking Cocktail Reception Begins with Drinks and Appetizers Sponsored by Taiwan Joint Commission of Precision Medicine

Morning Coffee and Breakfast Pastries and Networking

Exosome Engineering for Protein Delivery Via Optogenetic Approach
Chulhee Choi, Professor and Chair, BioMedical Imaging Center, Korea Advanced Institute of Science and Technology (KAIST), President, ILIAS Biologics Incorporated, Korea South

In this presentation, an opto-genetically engineered exosome system, named ‘exosomes for protein loading via optically reversible protein–protein interaction” (EXPLOR) that can deliver soluble proteins into the cytosol via controlled, reversible protein–protein interactions (PPI) will be introduced. By integrating a reversible PPI module controlled by blue light with the endogenous process of exosome biogenesis, cargo proteins can be loaded into newly generated exosomes. Treatment with protein-loaded EXPLORs was shown to significantly increase intracellular levels of cargo proteins and their function in recipient cells in both a time- and dose-dependent manner. Previously, it has been shown to delivery mCherry, Cre enzyme, Bax, and Super repressor I?B proteins as functional proteins in the target cells and in vivo. In this presentation, the results for follow-up studies will also be discussed.

Mid Morning Coffee and Tea Break

Networking Lunch

Carboxypeptidase E: A Cancer Biomarker in Circulating Exosomes
Y. Peng Loh, Chief and Senior Investigator, Section on Cellular Neurobiology, Eunice Kennedy Shriver National Institute of Child health and Human Development, National Institutes of Health (NIH), United States of America

Tumor recurrence and metastasis are the major causes of death in cancer patients. Biomarkers that can predict tumor recurrence in cancer patients who are in the early pathology stage and able to receive curative resection will greatly improve survival. To facilitate early cancer diagnosis and as a companion diagnostic to follow the efficacy of the therapy, such biomarkers should be detected not only in resected tumors, but also in serum to provide a non-invasive assay. Accumulating evidence suggest that carboxypeptidase E (CPE), could serve as a biomarker for predicting recurrence and survival in certain cancers, although lots remains to be studied to check on its validity and reproducibility. Overexpression of carboxypeptidase E (CPE) mRNA is common in many different human cancer types including metastatic cervical cancer, renal (clear cell) carcinoma, Ewing sarcoma, glioblastoma and various types of astrocytomas and oligodendrogliomas [1]. Indeed, CPE mRNA overexpression in resected tumors was significantly correlated with poor prognosis in early-stage HCC [2] and cervical [3] cancer. Since tumor cells release exosomes which contain mRNAs and proteins that mirror their parent tumor cells, and are readily accessible in nearly all biological fluids, we have developed a method to measure CPE mRNA in exosomes. CPE mRNA was elevated in exosomes isolated from cell culture media of liver, prostate, ovarian, glioblastoma and colorectal cancer cell lines with high compared to low metastatic potential. There was no correlation between the size and numbers of secreted exosomes and the metastatic potential of the cancer cells. Our preliminary studies indicate that CPE mRNA is present and elevated in serum-derived exosomes of many of the patients with different types of cancers including cervical, ovarian and breast cancer, compared to normal controls. Thus circulating exosomal CPE mRNA is a potentially useful biomarker for diagnosis of cancer and early detection of recurrence post-surgery, although much more work lies ahead to develop this as a valid test.

Single-use Biomanufacturing of Extracellular Vesicle Reference Materials
William Whitford, Strategic Solutions Leader, GE Healthcare, United States of America

Interest in microvesicles, exosomes and oncosomes is growing. Applications include 1) vectors of research or therapeutic cargo, 2) agents of intercellular communication from stems cells to terminally differentiated tissue  to the entire microbiome and 3) support of clinical diagnostics.  There are ongoing efforts to standardize clinically applied vesicle assays and therapeutic cargo vehicles.  Reference materials, controls, and performance standards need to be defined for quality assurance in such applications.  Certified reference materials (such as from the NIST or ATCC) and secondary materials may need to be generated for CLEA regulated diagnostic or therapeutic activities.  Sponsors often have their choice of cell platforms, production formats and culture modes for vesicle product and process development. However, commercial success can be dependent upon the discovery of scalable technologies that can produce very large amounts of sufficiently pure vesicles in a robust, compliant and cost-effective manner in a cGMP environment.  In biopharmaceuticals, single-use processing  is demonstrating heightened process efficiency and flexibility, reduced contamination risks, lower initial facility investment costs and operating costs, educed classifications and utilities.  Continuous biomanufacturing additionally promises a reduction in product microheterogeneity;  construction costs and schedule extent; manufacturing suite area and  classification.  The value of single-use implemented continuous biomanufacturing with chemically defined animal product-free materials will be reviewed.

Analysis of Single Cells Released by a Detector Designed for in vivo Catching of Circulating Tumor Cells
Peter Sedlmayr, Associate Professor, Medical University of Graz, Austria, Austria

We evaluated the Catch-and-Release Detector, an anti-EPCAM-coated wire designed for in-vivo catching of circulating tumor cells and subsequent in vitro release. DNA quality keeps stable as shown by array-CHG and NGS, indicating applicability for single CTC genomic analysis.

Circulating microRNA as Prognostic Marker in Myelodysplastic Syndromes
Zhuang Zuo, Associate Professor, Department of Hematopathology, The University of Texas MD Anderson Cancer Center, United States of America

Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal bone marrow hematopoietic stem cell disorders characterized by ineffective hematopoiesis and peripheral cytopenias. Molecular markers, however, are not integrated into currently used prognostic systems. The goal of this study is to identify plasma microRNAs useful for classification and risk stratification in MDS. We applied a novel, high-throughput digital quantification technology (NanoString) to profile microRNA expression in plasma samples of MDS patients. We correlated these results with overall survival and identified a 7-microRNA signature as an independent predictor of survival. These results validate the utility of circulating microRNA levels as noninvasive biomarkers that can inform the management of MDS patients. Our findings also shed light on interactions of gene regulation pathways likely involved in the pathogenesis of MDS.

Close of Day 2 of the Main Conference

Start of Training Course on Microfluidics/Lab-on-a-Chip Technologies and Liquid Biopsies [Separate Registration Required]