Conference Opens and Welcoming Speech by Associate Dean of National Taiwan University
Tsai-Kun Li, Associate Dean & Professor, National Taiwan University College of Medicine; Chief Executive Officer, Center for Genomic Medicine, National Taiwan University, Taiwan

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

Plasma Extracellular Vesicle Content for Diagnosis and Prognosis of 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. Atherosclerosis is the underlying syndrome for CVD and atherosclerotic plaques are detectable already in teenagers. Therefore for cardiovascular disease, we have to determine who is at high risk for a cardiovascular event in a high-risk background of the atherosclerotic syndrome without the possibility of taking a plaque biopsy. The risk for Cardiovascular events is region-dependent so tailored plasma-based biomarkers are essential to guide prevention and treatment of billions. Collecting plasma extracellular vesicles is like taking a liquid biopsy from the pathological tissue that can be used for diagnosis and prognosis of the disease. We show that plasma extracellular vesicle protein content especially when collected from different plasma subfractions can be used as an accurate source for diagnosis and prognosis of cardiovascular disease.

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, United States of America

The NIH Extracellular RNA Communication Program: exRNA for Therapy and Biomarker Development
Danilo Tagle, Director, Office of Special Initiatives, National Center for Advancing Translational Sciences at the NIH (NCATS), United States of America

Extracellular RNA (exRNA) can act as an endocrine signal to alter the phenotypes of target cells, and represents a novel paradigm in intercellular signaling. RNAseq analyses have identified a diverse population of exRNA that have been linked to regulation of the epigenome which could have profound implications for a wide range of physiologic and pathologic processes. The NIH Extracellular RNA Communication program supports research in exRNA-based biomarker and therapy development, as well as understanding fundamental principles of their biogenesis, distribution, uptake and function.

NanoVelcro-Embedded Microchips for Detection and Characterization of Circulating Tumor Cells
Hsian-Rong Tseng, Professor, Crump Institute for Molecular Imaging, California NanoSystems Institute, University of California-Los Angeles, United States of America

Inspired by the nanoscale interactions observed in the tissue microenvironment, our research team at UCLA pioneered a unique concept of “NanoVelcro” cell-affinity substrates, in which CTC capture agent-coated nanostructured substrates were utilized to immobilize CTCs with high efficiency. The working mechanism of NanoVelcro cell-affinity substrates mimics that of VelcroTM – when the two fabric strips of a Velcro fastener are pressed together, tangling between the hairy surfaces on two strips leads to strong binding.  Through continuous evolution, 3 generations of NanoVelcro CTC Chips have been established to achieve different clinical utilities. My presentation will summarize the continuous evolution of NanoVelcro CTC Assays from the emerge of the original idea all the way to their applications in cancer research and diagnosis. We envision that NanoVelcro CTC Assays will lead the way for powerful and cost-efficient diagnostic platforms for researchers to better understand underlying disease mechanisms and for physicians to monitor real-time disease progression.

Microfluidics for Liquid Biopsy & Personalized Medicine
Chwee Teck Lim, NUS Society Chair Professor, Department of Biomedical Engineering, Institute for Health Innovation & Technology (iHealthtech), Mechanobiology Institute, National University of Singapore, Singapore

Capturing of circulating Tumor Cells (CTCs) from the bloodstream of patients (a.k.a. liquid biopsy) may lead to better cancer detection and personalized medicine.  Using our developed microfluidic biochips, we not only showed successful detection and retrieval of CTCs, but also demonstrated how molecular analysis of each single CTC can enable personalized treatment by the detection of any actionable or druggable mutation.

Circulating Tumor Cells: The Promise and Reality of the Liquid Biopsy
Minetta Liu, Associate Professor and Chair, Oncology Research, Mayo Clinic, United States of America

The presentation objectives are as follows:  To discuss the potential impact of CTC molecular characterization in clinical practice. To review CTC isolation platforms in the context of demonstrating clinical utility as a liquid biopsy.

ExoScreen as a Novel Liquid Biopsy Platform for Detecting Cancer-Specific Exosomes
Takahiro Ochiya, Professor, Department of Molecular and Cellular Medicine, Tokyo Medical University, Japan

A highly sensitive and rapid analytical technique for profiling circulating EVs/exosomes directly from blood samples of patients has also been established that uses two types of antibodies to capture two specific antigens residing on EVs and can be detected by photosensitizer beads. Using this system, higher signal detecting CD147 and CD9 double-positive EVs were observed in serum from colorectal cancer (CRC) patients compared with that in serum from healthy donors. A part of sera from CRC patients including stage I and II were positive for CD147, although both CEA and CA19-9 positive serum could not be found in those patients. On the basis of these results, this new ExoScreen technology may enable the effective diagnosis of cancer.

EFIRM Liquid Biopsy (eLB)
David Wong, Felix and Mildred Yip Endowed Chair in Dentistry; Director for UCLA Center for Oral/Head & Neck Oncology Research, University of California-Los Angeles, United States of America

The advent of personalized medicine employing molecular targeted therapies has markedly changed the treatment of cancer in the past 10 years. Although tumor tissue biopsy-based genotyping is the current clinical practice for guiding clinical management, biopsy procedures can result in significant morbidity, limiting sampling to static snapshots which are further limited in scope by the inherent sampling bias of the analysis itself. To overcome these issues, technologies are needed for rapid, cost-effective, and noninvasive identification of biomarkers at various time points during the course of disease. Liquid biopsy is a rapidly emerging field to address this unmet clinical need as diagnostics based on cell-free circulating tumor DNA (ctDNA) can be a surrogate for the entire tumor genome. The use of ctDNA via liquid biopsy will facilitate analysis of tumor genomics that is urgently needed for molecular targeted therapy. Currently, most targeted approaches are based on PCR and/or next generation sequencing (NGS) for liquid biopsy applications with performance concordance in the 60-80% range with biopsy-based genotyping. We have developed a liquid biopsy technology “Electric Field Induced Release and Measurement (EFIRM)- Liquid Biopsy (eLB)” provides the most accurate detection that can assist clinical treatment decisions for the most common subtype of lung cancer, non-small cell lung cancer (NSCLC), with tyrosine kinase inhibitors (TKI) that can extend the disease progress free survival period of these patients. eLB detects actionable EGFR mutations in NSCLC patients with 100% concordance with biopsy-based genotyping and outperforms current technologies (digital droplet PCR [ddPCR] and NGS) for liquid biopsy. eLB is minimally/ non-invasive, rapid, inexpensive and self-contained permitting the detection of the most common EGFR gene mutations that are treatable with TKI such as Gefitinib or Erlotinib to effectively extend the progression free survival of lung cancer p

Rare Cell Diagnostics
Daniel Chiu, A. Bruce Montgomery Professor of Chemistry, University of Washington, United States of America

This presentation will describe advances we have made with the eDAR platform for the isolation of rare cells, with emphasis on employing eDAR for the analysis of circulating tumor cells.

Limitations and Challenges in MicroGenomics -- What we can Learn from Single-Cell and Exosome Expression Profiling
Michael Pfaffl, Professor, Technical University of Munich, Germany

So far quantitative real-time RT-PCR is the ‘Gold standard’ in molecular and clinical diagnostics, due to its high sensitivity, good reproducibility, and the wide dynamic quantification range. It is the most sensitive method for the quantification of mRNA/microRNA expression levels, in particular for extreme low abundant transcripts in tissue samples or liquid biopsies with limited RNA concentrations, e.g. single-cells, microvesicles or exosomes. Today the identification of novel and valid ‘biomarker signatures’ are in the focus. Therefore RNA Sequencing is increasingly used in experimental and clinical samples, to get the ‘holistic view’ on the mRNA and small RNA transcriptome.

Multiplexed Detection of Circulating Tumor DNA using Microfluidic Surface Plasmon Resonance (SPR) Sensormetry
Sehyun Shin, Professor & Director, Nano-Biofluignostic Engineering Research Center, Korea University and Anam/Guro Hospital of Korea University, Korea South

Circulating tumor DNA (ctDNA) has been demonstrated as the most promising biomarker for non-invasive assessment of cancer as well as the most accurate predictor of cancer treatment responses. However, detection of ctDNA in blood has been faced with conventional technical limits and called for innovative technologies and methods.
Here, we present a stable and selective assay for multiplexing detection of epidermal growth factor receptor (EGFR) mutation of most common mutations in plasma using DNA-DNA hybridization and selective linker prove with a lab-made surface plasmon resonance (SPR) sensometry. We prepared wild type and mutant templates of E746-A750 deletion in exon 19. Linker DNAs coated on a sensor surface of SPR capture different quantities of two different DNA types. Due to characteristics of SPR, the whole process was monitored in real-time and completed within an hour. Quantification data of SPR was compared with result of peptide nucleic acid (PNA) – locked nucleic acid (LNA) clamping PCR method for EGFR mutation detection. This study as a proof of concept can be further expanded into multiplexing detection of major and known ctDNAs, which could provide a solution for clinical unmet needs in cancer treatment and early detection.

The Amoeboid Tumor Phenotype and Mechanisms of Tumor Migration and Conditioning of the Tumor Microenvironment
Michael Freeman, Vice Chair of Research, Director, Cancer Biology and Therapeutics, Cedars-Sinai Medical Center, United States of America

Closing Keynote: Multiplex Detection of Extremely Rare Mutant Sequences Associated with Cancer
Fred Kramer, Professor of Microbiology, Biochemistry & Molecular Genetics, New Jersey Medical School Rutgers University, United States of America

Extraordinarily specific “SuperSelective” PCR primers enable the simultaneous quantitation of extremely rare mutations that occur in different cells, even though they are located within the same or adjacent codons.  Consequently, multiplex assays for the early detection of mutations associated with cancer should enable therapy to be guided by the results of periodic “liquid biopsies” that analyze DNA fragments present in plasma samples.