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SELECTBIO Conferences Liquid Biopsies and Minimally-Invasive Diagnostics 2016

Liquid Biopsies and Minimally-Invasive Diagnostics 2016 Agenda

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Thursday, 29 September 2016


Conference Registration, Conference Materials Pick-Up, Morning Coffee and Breakfast Pastries in the Exhibit Hall

Session Title: Emerging Trends in Liquid Biopsies for Cancer


Walter KochKeynote Presentation

Liquid Biopsy Tests Move Towards Routine Patient Care and Management
Walter Koch, Vice President, Roche Molecular Systems, United States of America

Over the past decade several tissue-based companion diagnostic assays have gained FDA approval in conjunction with targeted cancer therapies, resulting in improved treatment options for many cancer patients.  Despite various international guidelines, not all patients are tested, due in part to limited tissue availability, or health status limits on acquiring a re-biopsy sample.  This has spurred development of so-called liquid biopsy tests which use body fluids such as plasma to ascertain tumor mutational status in circulating cell free DNA originating from tumors, or circulating tumor cells.  On June 1, 2016 the FDA-approved the cobas® EGFR Mutation Test v2 real-time PCR test for the qualitative detection of defined mutations of the epidermal growth factor receptor (EGFR) gene in non-small cell lung cancer (NSCLC) patients. For the first time the test can be used with both formalin-fixed paraffin-embedded tumor tissue (FFPET) as well as circulating-free tumor DNA (cfDNA) from plasma derived from EDTA anti-coagulated peripheral whole blood.  The test is indicated as a companion diagnostic to aid in selecting NSCLC patients for treatment with the targeted therapies including TARCEVA®. This milestone heralds the first of many to come that will provide additional opportunities for cancer patients to be evaluated with blood samples for treatment with targeted therapies.  Analogous to HIV viral load testing, further clinical development will likely extend the use of such blood tests to allow monitoring of initial drug response as well as disease progression and/or development of specific resistance mutations.  Collectively these advances promise to provide important clinical information not easily obtained by repeat tissue biopsy approaches.


Minetta LiuKeynote Presentation

Promise of Circulating Tumor Cell and Cell-Free DNA Assays in the Oncology Clinic
Minetta Liu, Associate Professor and Chair, Oncology Research, Mayo Clinic, United States of America


Michael HellerKeynote Presentation

Cell Free DNA, RNA and Exosome Liquid Biopsy Cancer Diagnostics
Michael Heller, Professor, Dept Bioengineering, University of California-San Diego, United States of America

Circulating cell free (ccf) DNA, RNA and exosomes are now considered important biomarkers for liquid biopsy cancer diagnostics, and also hold great promise for early cancer detection. Nevertheless, the isolation of these biomarkers from patient samples requires relatively complex, time consuming and expensive procedures which greatly limits their practical use for most cancer diagnostic applications. New AC dielectrophoretic (DEP) microarray/chip devices now allow 15-20-minute isolation of cancer related ccf-DNA, RNA and exosome biomarkers from 20-50ul of blood, plasma or serum. After isolation of the biomarkers, specific fluorescent dyes can be used first to simultaneously detect the different biomarker levels directly on the chip (in-situ). In a subsequent step, immunofluorescent analysis can be carried out to identify specific protein biomarkers on the exosomes. Finally, the ccf-DNA and RNA (mRNAs and miRNAs release from the exosomes) can be eluted from the DEP chip, and PCR and sequencing analysis carried out to identify the cancer-related point mutations and other polymorphisms, as well as to further verify the tissue origin of the biomarkers. In the case of our Chronic Lymphocytic Leukemia clinical studies, final PCR and DNA sequencing results for the CLL related ccf-DNA isolated by DEP were found to be exactly comparable to two much more complex and time consuming “gold standard” procedures. In the case of glioblastoma exosomes isolated from plasma, exosome-specific surface and interior proteins CD63 and TSG101 could be detected by immunofluorescence, and mutated EGFRvlll mRNA was detected by RT-PCR. Finally, the exosomal related protein biomarker Glypican-1 could be isolated from pancreatic cancer patient plasma samples by DEP and detected on-chip by immunofluorescence. Thus, DEP represents a powerful new minimally invasive technology for cancer diagnostics that is particularly well suited for the rapid isolation of cell free nucleic acid and exosomes.


Dominique PV de KleijnKeynote Presentation

Diagnosis of Ischemic Heart Disease Using Plasma Extracellular Vesicle Sub-fractions
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. Ischemic heart disease (IHD) comprises 3 entities: stable coronary artery disease (SCAD), unstable angina (UA) and myocardial infarction (MI). Because IHD is associated with an increased risk of adverse clinical events such as heart failure and death, early recognition of IHD is of utmost importance. However, to diagnose IHD is challenging, as many patients present with atypical symptoms. It is known that women have a different symptom sensation than men. Troponins are the main diagnostic tool for detection of MI. Blood biomarkers for SCAD (typically causing stable angina) and UA, however, are not available. These diagnoses frequently require hospital visits/admissions for time-consuming and costly (non)invasive tests. Using 4 plasma fractions, we show that the plasma extracellular vesicle content in these 4 fractions can be used as an accurate source for early diagnosis of SCAD and UA.


Coffee Break and Networking in the Exhibit Hall: Visit Exhibitors and Poster Viewing


NGS Libraries From Cell-free DNA Containing Molecular Tags Prepared with ThruPLEX® Technology Improve Ability to Detect Rare Alleles
Karl Hecker, Vice President of Product Development, Rubicon Genomics, Inc., United States of America

Next Generation Sequencing (NGS) of cell-free DNA (cfDNA) enables non-invasive access to genetic information from liquid biopsies. Key to identifying rare genetic mutations is improved sequencing accuracy and the ability to distinguish between biological and PCR duplicates. Rubicon Genomics’ ThruPLEX Plasma-seq was developed specifically for cell-free DNA.


Combining CTC and ctDNA Platforms from the Same Clinical Samples to Expand Liquid Biopsy Biomarker Content
Lyle Arnold, Chief Scientific Officer, Biocept, United States of America

Liquid biopsies offer the opportunity to interrogate a number of different target sample types, including ctDNA and CTCs. At Biocept both ctDNA and CTCs are used for identifying medically actionable biomarkers. The combination of these technologies has enabled the clinical validation of an array of biomarkers from single blood samples. These include mutational analysis for EGFR, KRAS, and BRAF, as well as FISH and protein analysis for HER2, FGFR1, MET, ALK, ROS1, ER, PR, AR and PDL-1 across a range of cancer types.  In addition, CTCs can be isolated from the microchannel and interrogated using other means, including NGS.


Covaris, Inc.Technology Spotlight:
truXTRAC cfDNA: The High Throughput, High Yield Adaptive Focused Acoustics™ (AFA)-Mediated Circulating Cell-Free DNA Extraction and Purification System
Hamid Khoja, Principal Scientist, Covaris, Inc.

The presentation will provide data illustrating the effects of an active extraction method of ccfDNA from plasma, and comparison data with the currently-used protocols.


Networking Lunch in the Exhibit Hall: Visit Exhibitors and Poster Viewing

Session Title: Analyte Classes for Liquid Biopsy Development -- Cell-free DNA (cfDNA), Circulating Tumor Cells (CTCs) and Extracellular Vesicles (EVs)


Longitudinal Whole Exome Investigation of Mutational Heterogeneity in Single CTCs in a Patient with Triple-Negative Breast Cancer
Eric Kaldjian, Chief Medical Officer, RareCyte, United States of America

We performed whole exome sequencing of single CTCs from a patient with metastatic triple-negative breast cancer to investigate the evolution of genetic heterogeneity during therapy.  CTCs were identified using the AccuCyte-CyteFinder system (RareCyte) and individually retrieved from microscope slides using the integrated CytePicker function.  Single cell whole genome amplication was performed followed by whole exome sequencing.   Computational biology tools were employed to analyze genomic DNA sequence from multiple CTCs, white blood cells and ctDNA from various time points. Genomic sequence alterations were observed to evolve over the course of therapy in individual CTCs.  These alterations appear to be generally consistent within CTC at a given time point.  The number of predicted deleterious and cancer driver mutations per CTC were observed to increase in frequency after effective treatments, suggesting that the number of such alterations may be associated with resistance to therapy.


Weian ZhaoKeynote Presentation

Digital Quantification of Biomarkers in Liquid Biopsies Using the Integrated Comprehensive Droplet Digital Detection (IC 3D) System
Weian Zhao, Associate Professor, Department of Biomedical Engineering, University of California-Irvine, United States of America

We will present a new platform technology that could detect rare blood biomarkers including infections, CTCs, miRNA and exosomes with extremely high sensitivity, specificity, throughput and minimal sample processing.


MMI Microscope-based Single Cell IsolationTechnology Spotlight:
Advances in Microscopy-Based Single Cell Isolation
Chris Wetzel, Director of Sales and Marketing, MMI Microscope-based Single Cell Isolation

The isolation of single cells can often present challenges depending on the properties of the cells and the surrounding medium. This presentation deals with state-of-the-art techniques how to isolate cells from tissue (Laser Micro Dissection) and liquid suspensions (automated Micro Capillary) including live cells and adherent cells.


AmirAli TalasazKeynote Presentation

Circulating Tumor DNA in Advanced-Stage Cancer Patients – Somatic Genomic Landscape and its Clinical Utility
AmirAli Talasaz, Co-Founder, President & COO, Guardant Health, United States of America

Next-generation sequencing (NGS) of circulating tumor DNA (ctDNA) enables non-invasive profiling of solid tumor cancers. Liquid biopsy studies to date have been limited to modest-size cohorts and case studies. Somatic genomic profiles of over 15,000 patients with advanced-stage clinical cancer were determined by a highly accurate, deep-coverage ctDNA NGS test targeting 70 genes (Guardant360). Frequencies of somatic ctDNA alterations per gene were compared to those previously described in tissue sequencing projects (e.g., TCGA). Accuracy of ctDNA sequencing (PPV) was assessed by comparing with matched tissue tests. Different classes of clinical outcome benefits have been observed by liquid biopsy by detecting actionable mutations in cases with tissue QNS or actionable mutations emerging at time of progression or under-genotyped tumors.


Coffee Break and Networking in the Exhibit Hall: Visit Exhibitors and Poster Viewing


Circulating Vesicles at the Nanoscale
Shivani Sharma, Associate Director, California NanoSystems Institute, California NanoSystems Institute, UCLA, United States of America

The role of nanotechnology in understanding the nanoscale structure, mechanics and biomolecular characteristics of circulating extracellular vesicles (EVs) will be discussed.


Steve SoperKeynote Presentation

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


Chamindie PunyadeeraKeynote Presentation

Could Precision Medicine be Tailor-Made for Metastatic Head and Neck Cancers
Chamindie Punyadeera, Associate Professor, Institute of Health and Biomedical Innovation, Queensland University of Technology, Australia


Lidong QinKeynote Presentation

Profile CTC Stiffness by Mechanical Separation Chips to Better Predict Breast Cancer Metastasis
Lidong Qin, Professor and CPRIT Scholar, Houston Methodist Research Institute, United States of America

Detection of Circulating Tumor Cells (CTCs) has emerged as a promising minimally invasive prognostic tool to stage cancer metastasis; however, the identification and characterization of CTCs require extremely sensitive and specialized analytical methods, as CTCs are rare, heterogeneous, and associated with a complexity of blood sample matrix. To combat the current CTC detection limitations, we plan to incorporate the filtration method based on a size exclusion capability of microfluidics chips with cocktails of biomarker-specific antibodies to stain and image CTCs. By observing the cell distribution on the chip based on CTCs’ capability in crossing micro-barrier arrays, we can profile the captured CTCs’ stiffness and correlate it to the cancer metastasis potential. We expect to provide a better cancer prognosis tool than counting cells.


Hsian-Rong TsengKeynote Presentation

Clinical Applications of NanoVelcro Rare-Cell Assays 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

Circulating tumor cell (CTC) is regarded as a liquid biopsy of tumor, allowing non-invasive, repetitive, and systemic sampling of disease. Although detecting and enumerating CTCs is of prognostic significance in metastatic cancer, it is conceivable that performing molecular and functional characterization on CTCs will reveal unprecedented insight into the pathogenic mechanisms driving lethal disease. Nanomaterial-embedded cancer diagnostic platforms, i.e., NanoVelcro CTC Assays represent a unique rare-cell sorting method that enables detection isolation, and characterization of CTCs in peripheral blood, providing an opportunity to noninvasively monitor disease progression in individual cancer patients. Over the past decade, a series of NanoVelcro CTC Assays has been demonstrated for exploring the full potential of CTCs as a clinical biomarker, including CTC enumeration, phenotyping, genotyping and expression profiling. In this presentation, Dr. Tseng will briefly introduce the development of three generations of NanoVelcro CTC Assays, and highlight the clinical applications of each generation for various types of solid cancers, including prostate cancer, pancreatic cancer, lung cancer, kidney cancer, liver cancer, and melanoma.


Cocktail Reception for All Conference Attendees: Enjoy Beer, Wine, Appetizers and Network with Fellow Delegates, Speakers, Exhibitors Pool-Side


Close of Day 1 of the Conference. Continue Networking in Downtown San Diego (Trolleys to the City are Available Right Behind the Conference Venue).

Friday, 30 September 2016


Morning Coffee, Breakfast Pastries and Networking in the Exhibit Hall


LATE-BREAKING PRESENTATION: A Platform For Characterization of Single Exosomes
David Freedman, Co-Founder & CEO, nanoView Diagnostics Inc., United States of America

Nanoview Diagnostics has developed a label-free visible-light microarray imaging technique that allows enumeration and sizing of individual nanovesicles captured on the sensor. The NVDX technology enables multiplexed (10-100’s) phenotyping of exosomes in a one-step assay direct-from-sample and can work with samples volumes as small as 5 µl. The NVDX exosome visualization technology is a direct-from-sample high-throughput technique than can improve standardization of exosome preparations and facilitate translation of exosome based liquid biopsies.


Fred KramerKeynote Presentation

SuperSelective Primers for Multiplex Real-time PCR Assays that assess the Abundance of Rare Mutations Associated with Cancer
Fred Kramer, Professor of Microbiology, Biochemistry & Molecular Genetics, New Jersey Medical School Rutgers University, United States of America

Real-time multiplex PCR assays are potentially the most rapid, most sensitive, and least expensive way to assess the abundance of mutant DNA fragments present in liquid biopsies; provided that a way is found to selectively amplify rare mutant fragments without amplifying abundant related wild-type fragments; and provided that the amplicons generated from different mutants that occur in the same or an adjacent codon are prevented from forming heteroduplexes that interfere with exponential amplification, obscuring the threshold values of the rarer mutants.  “SuperSelective” PCR primers, due to their unique design, are extraordinarily specific, able to selectively initiate the synthesis of amplicons on ten mutant DNA fragments in the presence of 1,000,000 wild-type DNA fragments, even though the only difference between the mutant and the wild-type is a single-nucleotide polymorphism.  Each SuperSelective primer that is specific for a particular mutation possesses a unique 5' tag sequence that is incorporated into the resulting amplicons and detected in real-time by differently colored molecular beacon probes.  Each SuperSelective primer specific for a particular mutation also possesses a unique “bridge” sequence that assures that each primer only copies its intended amplicon, and that creates a single-stranded bubble in heteroduplexes that enables the rarest amplicons to be independently exponentially amplified.  And finally, the inclusion of primers for a wild-type reference gene fragment, enables the abundance of each type of mutant DNA fragment to be assessed (without measuring the amount of DNA in the sample) by determining the difference between its threshold value and the threshold value of the reference gene.

Session Title: Liquid Biopsies in Disease Classes: Cancer and Looking Beyond


Liquid Biopsies in the Management of Cancer
James Han, Director, Bioinformatics, Genomic Health, United States of America

Liquid biopsies can be used to monitor tumor dynamics including recurrence or to profile individual genetic and genomic markers that are informative of treatment options.  Together, these complementary approaches provide precision solutions to help manage disease along the patient cancer journey.  These also call for different development, analytical and clinical validation strategies, as well as demonstration of clinical utility.  As these assays become more routine and move toward standard of care, it will be important to provide accurate and relevant test performance information.  I will discuss the role of standards to ensure quality and allow appropriate interpretation of test results.


Extracellular RNAs to Monitor the CNS: An Examination of Several Biofluids and Isolation Strategies
Kendall Van Keuren-Jensen, Professor and Deputy Director, Translational Genomics Research Institute, United States of America

We explore several different biofluids across several different diseases and injuries of the central nervous system (CNS).


Extracellular RNAs as Biomarkers for Placental Function and Dysfunction
Louise Laurent, Associate Professor, University of California-San Diego, United States of America

Placental dysfunction is responsible for a large fraction of the morbidity and mortality seen in pregnancy, and includes preeclampsia and fetal growth restriction. Although the clinical manifestations of these complications arise in the second half of pregnancy, they stem from abnormal placental development during the first trimester. Therefore, we hypothesize that there are biomarkers that can be evaluated early in pregnancy that will predict later onset of clinical signs and symptoms. Building upon the knowledge that placental RNAs can be detected in the serum of pregnant women, we have used next-generation sequencing of extracellular RNA to identify candidate miRNAs for the prediction of placental dysfunction. Here, we will discuss the techniques used for generation and analysis of the data, and present results supporting the discovery and validation of these biomarkers.


Coffee Break and Networking in the Exhibit Hall: Visit Exhibitors and Poster Viewing


John SninskyMarica GrskovicKeynote Presentation

Cell-Free DNA: Potential Pan-Organ Marker for Organ Transplantation
John Sninsky, Chief Scientific Officer, CareDx
Marica Grskovic, , CareDx, United States of America

Patients who have undergone organ transplants require frequent monitoring to evaluate the organ’s graft status and to modulate a complex regimen of immunosuppressive medications to address short and long term clinical needs. Sub-optimal dosing and the threat of allograft rejection must be balanced with excessive dosing and increased risks of infections and cancer. A significant unmet medical need exists for clinical diagnostic tools to permit surveillance management of transplant patients and improve the long-term outcomes of immunosuppressive therapy. Cell-free DNA (cfDNA) has been described as a biomarker for prenatal testing, cancer, and organ transplantation, each of which presents different clinical and technological challenges. cfDNA circulating in the plasma of transplant patients represents a mixture of residual nucleosomal-protected genomic regions released from dying cells of allograft (“transgenome”) and recipient cfDNA. The genomes of the organ donor and allograft recipient are distinguishable by next generation sequencing. Longitudinal samples from heart and kidney transplant patients showed significantly higher donor-derived cfDNA levels prior to biopsy-confirmed rejection which were reduced following adjustments to immunosuppressive therapy. The cfDNA compartment also harbors information about specific infectious diseases and somatic mutations associated with cancer development.  The role of Big Data in tracking organ transplantation outcomes will be discussed.


Monitoring Rejection and Infection through Analyses of Circulating Cell-free (cf) DNA
Iwijn De Vlaminck, Robert N. Noyce Assistant Professor in Life Science and Technology Meinig School of Biomedical Engineering, Cornell University, United States of America

This talk will cover applications of cell-free DNA in the diagnosis of rejection in solid-organ transplantation, as well as applications of cell-free DNA monitoring in the broad, hypothesis-free monitoring of infection.


Therapeutic and Diagnostic Targeting of Exosomes in Cancer and Infectious Disease
James Joyce, Chairman & CEO, Aethlon Medical, Inc., United States of America

Discussion of lectin-affinity techniques to eliminate disease-promoting exosomes from circulation and the isolation of exosomal biomarkers from bodily fluids.


Networking Lunch in the Exhibit Hall: Visit Exhibitors and Poster Viewing


Clark ChenKeynote Presentation

CSF microRNA Profile as a Liquid Biopsy Platform to Assess Glioblastoma Tumor Burden
Clark Chen, Co-Director, University of California-San Diego, United States of America

Glioblastoma is the most common form of primary brain neoplasm and remains one of the deadliest of human cancers.  Clinical care for glioblastoma patients is largely limited by the absence of biomarker for assessment of tumor burden or therapeutic response. The discovery that glioblastoma cells secrete extra-cellular vesicles (EVs) containing tumor specific genetic material has opened a new platform for biomarker development.  These EVs transgress anatomic compartments and can be detected in clinical bio-fluids. The EVs shelter the tumor-specific genetic material from the extracellular environment that is replete with RNAses and preserve the integrity of these materials. Importantly, the genetic materials within EVs appear highly enriched for RNAs in the size range of microRNAs (miRNAs). The talk will focus on using cerebrospinal fluid (CSF) derived exosomal miRNA signature as a liquid biopsy platform for assessing glioblastoma tumor burden. Issues pertaining to vesicle quantitation, normalization, optimal specimen storage condition, and influence of microenvironment will be discussed.

Session Title: Emerging Technology Trends and Themes in the Liquid Biopsy Field


Celsee DiagnosticsTechnology Spotlight:
Automated, Sensitive Microfluidic Device for CTC Capturing and Characterization
Grant Howes, Vice President, Commercial Operations , Celsee Diagnostics

The Celsee Diagnostics’ novel microfluidic technology captures and characterizes CTCs from metastatic cancer patients’ whole blood samples based on size and deformability. The design enables downstream characterization of CTCs, including IHC, FISH, PCR and NGS.

This presentation will overview the technology, demonstrate the 85% capture efficiency and discuss the use of the Celsee PREP100 to enrich and retrieve CTCs for downstream applications. The Celsee PREP100 device uses a microfluidic chip, which has approximately 56,400 individual cell capturing wells. Each well ensures that smaller blood cells, such as red blood cells and most leukocytes, escape while larger CTCs are captured.

Downstream analysis of the captured CTCs using immuno-staining, FISH assays, RTPCR and NGS will be highlighted.


Coffee Break in Exhibit Hall


Protein Profiling of Tumour-derived Exosomes for Plasma Biomarker Discovery in Non-Small Cell Lung Cancer
Lingzhi Wang, Senior Research Scientist, Ctr. for Translational Medicine, Adjunct Professor, National University of Singapore, Singapore

Lung cancers are often diagnosed at advanced stages with concomitant poor prognosis, making it the leading cause of cancer mortality worldwide. Although low-dose CT have shown to be useful in detecting small abnormalities in the lung and associated with a 20% reduction in lung cancer-specific mortality, they have also been found to have high false positive rate of detection (96.4%) in a recent U.S.A. National screening trial. It is thus important to identify novel minimally-invasive lung cancer biomarkers. Circulating exosomes are important resources for cancer biomarker discovery because exosomes can reflect the cell’s RNA and protein contents from which they are secreted.


Y. Peng LohKeynote Presentation

Carboxypeptidase E: A Biomarker for Detection of Cancer 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), along with its N-terminal truncated derivative, could serve as a powerful prognostic biomarker for predicting recurrence and survival. Over-expression 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.


Nucleic Acid Mass Spectrometry in Minimally-Invasive Sensitive Diagnostics
Anders Nygren, Senior Director, Research and Development, Agena Biosciences, United States of America


Raghu KalluriKeynote Presentation

Exploiting the Biology of Exosomes for the Diagnosis and Treatment of Pancreatic Cancer
Raghu Kalluri, Professor and Chairman, Department of Cancer Biology, University of Texas MD Anderson Cancer Center, United States of America


Sehyun ShinKeynote Presentation

Novel Multiplexing and Precision Sensing Approaches for Liquid Biopsy
Sehyun Shin, Professor & Director, Nano-Biofluignostic Engineering Research Center, Korea University and Anam/Guro Hospital of Korea University, Korea South

Every year, there are about 899,000 new cases and 260,000 mortalities, comprising 6 percent of all cancer deaths globally. Cancer detection is a critical approach for preventing cancer deaths because cases caught early are often more treatable. The development of non-invasive methods to detect and monitor tumors remains a major challenge in oncology. Liquid biopsy such as circulating tumor cells (CTCs), exosomes and circulating tumor DNA (ctDNA) has the potential to provide information about cancers without invasive biopsy. It has been rapidly developing since it yields minimum invasiveness, easy repetitive sampling, and whole mutation representation. These circulating biomarkers can be easily obtained from biofluids such as saliva and blood. Although PCR and NGS (Next Generation Sequencing) based ctDNA detection methods have been introduced, there are still unmet needs to detect ctDNA for clinical applications. We presented a clampSPR (closed-loop amplification PCR-based Surface Plasmon Resonance) sensometry for detecting ctDNAs with multiplexing assay; detect biomolecules with label-free analysis via changes in the refractive index on the sensing film of the SPR sensor in real time. The proposed clampSPR system is similar to real time PCR. After a sample is loaded to the system, it is circulated through a closed-loop channel with passing three different temperature zones by an innovative microfluidic pumping system. Target genes in a sample are amplified as two folds per circulation. Furthermore, encoded hydrogel microparticles were added for multiplexing. This system is fast as 30 minutes for 40 cycles. In addition, it is portable, minimize the sample volume with low cost. Conclusively, our novel approaches can apply to clinical diagnosis of cancer with several advantages such as high sensitivity and multiplexing.


Dolores Di VizioKeynote Presentation

Large Oncosomes: New Frontiers for Cell-to-Cell Communication in Cancer
Dolores Di Vizio, Professor, Cedars Sinai Medical Center, United States of America

Extracellular vesicles (EVs) are important mediators of intercellular mechanisms as they can shuttle from one cell to another a reservoir of functional molecules (bioactive proteins, nucleic acids and lipids). EVs are highly heterogeneous and differ by size, composition and function. As a common characteristic, they all are surrounded by a lipid bilayer and can act in the proximity of the cell or at distance. Given the abundance of cancer-derived molecules that can be found in each particle, EVs are being recognized as appealing biomarkers of diagnosis and prognosis. Because these molecules are functional, EV profiling has also the potential to identify therapeutic targets in the personalized medicine effort. Our team recently reported that highly metastatic cells export 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 correlates with advanced disease in mouse models and human subjects. We show that DNA and RNA-Sequencing can identify tumor specific alterations in large oncosomes circulating in mice and patient plasma. Large-scale profile analyses demonstrate that large oncosomes represent a novel population of EVs enriched in tumor-derived molecules. Large oncosomes are thus valuable candidates for new biomarker profiles to be developed using tissue- and blood-based assays in combination.


Role of Exosomic microRNAs in the Biology of the Tumor Microenvironment
Muller Fabbri, Assistant Professor, University of Southern California, United States of America

Exosomes and other Extracellular Vesicles are gaining increasing interest for their role in shaping the biology of the tumor microenvironment. Recently, we showed that microRNAs contained in exosomes released by cancer cells are able to bind to Toll-like Receptors in surrounding Tumor-Associated Macrophages, leading to increased cancer growth and resistance to chemotherapy. This novel mechanism of action of microRNAs reveals a fascinating new aspect of the biology of the Tumor Microenvironment and identified exosomic microRNAs as potential new molecular target for anti-cancer therapy.


Isolation of Sub-populations of Prostate Cancer Cells Using Inertial Microfluidics
Ian Papautsky, Richard and Loan Hill Professor of Bioengineering, Co-Director, NSF Center for Advanced Design & Manufacturing of Integrated Microfluidics, University of Illinois at Chicago, United States of America

This work successfully demonstrates the use of an inertial microfluidic device as a laboratory tool for sorting sub-populations of circulating tumor cells (CTCs), and shows that the effects of fluidic shear on cell viability, functionality and “stemness” are negligible.


Close of Conference.

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