Cambridge Healthtech Institute’s 6th Annual

Enabling Technologies for Circulating Biomarkers

Improving Assays for Clinical Use

7-8 May 2019



Technologies to detect and analyze circulating cell-free DNA, CTCs, and other circulating biomarkers are continuing to develop and advance, but proper validation must occur before they can be used for routine clinical use. Cambridge Healthtech Institute’s 6th Annual Enabling Technologies for Circulating Biomarkers conference will examine the latest advances in such technologies and their path to clinical use. This year’s program will also focus on pre-analytical challenges: sample prep, detection, extraction, isolation, and characterisation. Special attention will be paid to other types of circulating biomarkers and sample types, and the lessons we can learn for use in cfDNA technologies.

Final Agenda

Recommended Short Course* Monday 6 May | 13:30 - 17:00

SC2: Biomarkers in Liquid Biopsy: CTCs, ctDNA and Exosomes

Lorena Diéguez, PhD, Group Leader, Department of Life Sciences, Nano4Health Unit, Medical Devices Research Group, International Iberian Nanotechnology Laboratory, Portugal

Roberto Piñeiro Cid, PhD, Cancer Modeling Lab, Instituto de Investigación Sanitaria de Santiago de Compostela- Roche-Chus Joint Unit, Spain

Biomarkers for early disease detection, therapeutic efficacy monitoring and outcome prediction are the key to precision medicine. Liquid Biopsy studies disease biomarkers in body fluids and can be paramount for precision medicine in cancer. The analysis of biomarkers in peripheral blood improves cancer diagnosis and treatment success. This course will give you a comprehensive overview and update on the established biomarkers, available technologies and clinical applications of liquid biopsy.

*Separate registration required.

TUESDAY 7 MAY

08:00 Registration and Morning Coffee

IMPLEMENTATION OF CIRCULATING BIOMARKER TECHNOLOGIES

08:55 Organizer’s Opening Remarks

Kaitlin Kelleher, Conference Producer, Cambridge Healthtech Institute

09:00 Chairperson's Remarks

Jörg Tost, PhD, Director, Laboratory for Epigenetics & Environment, Centre National de Recherche en Génomique Humaine (CNRGH), CEA – Institut de Biologie Francois Jacob, France

09:05 Realising Value from Liquid Biopsy: Assuring Analytical Validity, Clinical Utility, and Cost-Effectiveness

Messenger_MichaelMichael Messenger, PhD, Head of Personalised Medicine and Health, Leeds Centre for Personalised Medicine and Health, University of Leeds, United Kingdom

Careful consideration of the evidence requirements of decision makers early on in the test evaluation pipeline can reduce the time and cost of research and improve the likelihood of a successful reimbursement outcome. This presentation will provide an overview of efficient approaches and protocols for evaluating liquid biopsies for rapid adoption and reimbursement, using examples from real case studies. Furthermore, it will highlight various frameworks used for quality appraising research evidence on analytical performance, clinical validity, clinical utility and cost-effectiveness, that are used by reviewers and Health Technology Assessment schemes to look at the risk of bias, inapplicability and irreproducibility.

09:35 Paving the Way to Use Liquid Biopsies in the Real World – IMI CANCER-IDs Learnings and Outlook

Thomas Schlange, PhD, Senior Biomarker Scientist, Global Biomarker Research, Bayer AG, Germany

The Innovative Medicines Initiative project CANCER-ID set out in 2015 to establish criteria for evaluating technologies in the liquid biopsy field. At the core of the 5-year program are harmonized protocols for clinical use of CTCs, ctDNAs and miRNAs, standards for benchmarking technologies and implementing these technologies in clinical studies. Learnings from CANCER-ID will be presented and an outlook of further joint stakeholder activities will be given.

VolitionRx 10:05 Nu.Q™-Complete: Comprehensive Nucleosome Profiling

Eccleston_MarkMark Eccleston, Business Development Director, Volition, Belgium

Volition has established the concept of epigenetic profiling of cell free, circulating nucleosomes using simple Nu.Q™ immunoassays. The Nucleosomics® platform is being used to develop blood tests for early detection of a range of cancers. We now present a broader analytical approach, employing immuno-capture, mass spectrometry, immunoassay and sequencing of constituent DNA to allow comprehensive analysis of circulating nucleosomes for serum or plasma. Clinical applications will be discussed.

 

 

10:35 Grand Opening Coffee Break in the Exhibit Hall with Poster Viewing

 

PRE-ANALYTICAL AND ANALYTICAL CONSIDERATIONS FOR CIRCULATING BIOMARKERS

11:15 Achieving Measurement Traceability for Molecular Diagnostics in Compliance with the IVDR

Devonshire_AlisonAlison Devonshire, PhD, Science Leader, Molecular and Cell Biology, LGC, United Kingdom

The new In Vitro Diagnostic Device Regulation (EU 2017/746) states that calibrators and control materials used in diagnostic tests “shall be assured through suitable reference measurement procedures and/or suitable reference materials of a higher metrological order;” however, there are a scarcity of reference materials and reference measurement procedures for the nucleic acid and cellular analytes commonly measured by liquid biopsy tests. This talk will discuss a framework for developing a reference system for circulating biomarkers and CTCs, and recent progress in developing digital PCR as a reference measurement procedure for gene quantification.

11:45 Obtaining Every Droplet of Information out of Each Liquid Biopsy Sample

Diéguez_LorenaLorena Diéguez, PhD, Group Leader, Department of Life Sciences, Nano4Health Unit, Medical Devices Research Group, International Iberian Nanotechnology Laboratory, Portugal

Why choose between CTCs, ctDNA or exosomes when you can have it all? What if the information provided by the different biomarkers is not redundant, but complementary? We use microfluidics and nanotechnology to extract the most of each sample. Phenotypic analysis of single CTCs and mutation analysis is done using microdroplets and Surface Enhanced Raman Spectroscopy.

12:15 Fundamentals for the Automatic Classification of Quantitative PCR Amplification Curves: A Biostatistical Approach

Roediger_StefanStefan Rödiger, PhD, Group Leader, Institute of Biotechnology, Brandenburg University of Technology Cottbus – Senftenberg, Germany

Quantitative polymerase chain reaction (qPCR) is a widely used bioanalytical method in human diagnostics. Until now, classifications (e.g., positive or negative reaction) are performed manually or by fixed threshold values. This classification is error-prone and based on the operator’s experience. We developed a scientific software, PCRedux, which calculates predictors (features) of amplification curves automatically. The predictors can be used for the automatic analysis of large data sets for machine learning applications.

Biorad 12:45 Urological Cancer: Taking Liquid Biopsy to the Clinic

Guldberg_PerPer Guldberg, Professor, Danish Cancer Society, Denmark

It has been known for decades that cancers of the genitourinary system shed cells into the urine. This paper discusses factors that should considered when developing clinically useful urine-based tests, including 1) diagnostic coverage of DNA biomarkers, 2) DNA detection methods, and 3) sample collection. Addressing these factors have led to the development of a urine-based test for bladder cancer, which has a very high sensitivity and therefore may replace the current golden standard, cystoscopy.

13:15 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

13:45 Session Break

PRE-ANALYTICAL AND ANALYTICAL CONSIDERATIONS FOR CIRCULATING BIOMARKERS (CONT.)

14:15 Chairperson's Remarks

Michael Messenger, PhD, Head of Personalised Medicine and Health, Leeds Centre for Personalised Medicine and Health, University of Leeds, United Kingdom

14:20 Analytical and Biological Variation of Analyses of Circulating Tumor DNA

Fredslund_Andersen_RikkeRikke Fredslund Andersen, PhD, Molecular Biologist, Department of Clinical Biochemistry, Vejle Hospital, Denmark

Serial analyses of circulating tumor DNA in cancer patients are being investigated for assessing treatment response or failure. It is important to determine if values have significantly increased or decreased compared to previous values. We have performed extensive validation of mutation and methylation analyses in cfDNA from colorectal cancer patients to determine analytical and biological variation. From these values the significant minimum change can be calculated.

14:50 Optimizing Liquid Biopsies for Clinical Use

Niels Pallisgaard, PhD, Molecular Biologist, Pathology, Roskilde University, Denmark

Sample tubes, sample size and handling, when to use pre-amplification, assay sensitivity and assay optimization as well as proper controls for sample and assay quality for liquid biopsies in a clinical setting will be discussed in this presentation.

TECHNOLOGIES FOR CELL-FREE DNA

15:20 KEYNOTE PRESENTATION: Novel Digital PCR and Mutation Enrichment Technologies for the Analysis of Clinically Relevant DNA Alterations in Liquid Biopsies

G. Mike Makrigiorgos, PhD, Professor, Dana Farber Cancer Institute and Harvard Medical School, United States

With the increasing interest in treatment assessment using liquid biopsy and circulating DNA, sensitive and multiplexed detection of tumor-derived alterations in blood are desirable. We provide novel forms of digital PCR, as well as mutation enrichment-based real time PCR methods that (a) enable several orders of magnitude improvement of detecting mutations or microsatellite instability than currently possible; (b) are highly multiplex-able; (c) reduce cost of analysis. Application in circulating DNA from clinical cancer samples will be presented.

15:50 Analysis of Mutations and Methylated Molecules in Cell-Free DNA Using Enhanced-ice-COLD-PCR Enrichment Combined with Next-Generation Sequencing

Tost_JorgJörg Tost, PhD, Director, Laboratory for Epigenetics & Environment, Centre National de Recherche en Génomique Humaine (CNRGH), CEA – Institut de Biologie Francois Jacob, France

Circulating cell-free DNA has great potential for non-invasive diagnostics, prediction and monitoring of treatment response. We have previously developed Enhanced-ice-COLD-PCR for the detection and sequence-based identification of mutations in mutation hotspots as well as methylated molecules. While the initial developed workflow used Pyrosequencing for a short time-to-results, we have now moved the assays to an NGS platform improving further signal/noise ratio, correlation with ddPCR results and providing detailed information on enriched molecules.

16:20 Refreshment Break in the Exhibit Hall with Poster Viewing

17:00 Breakout Discussions View Details

18:00 Welcome Reception in the Exhibit Hall with Poster Viewing

19:00 Close of Day

WEDNESDAY 8 MAY

08:00 Registration and Morning Coffee

SINGLE CELL ANALYSIS AND CELL-BASED DIAGNOSTICS

09:00 Chairperson's Remarks

An Hendrix, PhD, Assistant Professor, Laboratory of Experimental Cancer Research, Ghent University, The Netherlands

09:05 Biological and Technical Aspects of Single-Molecule Analysis in Liquid Biopsies

Stahlberg_AndersAnders Ståhlberg, Associate Professor, Sahlgrenska Cancer Center, Clinical Pathology and Genetics, University of Gothenburg, Sahlgrenska University Hospital, Sweden

Early detection of individual tumor cells and molecules is essential in cancer diagnostics. We have developed several approaches to analyze individual cells and molecules and applied them to various applications within cancer and beyond. Here, we present our experience of using ultrasensitive analysis from both a clinical and technical point of view. Data from different tumor entities will be shown.

09:35 Inertial Microfluidics for High-Throughput Isolation of Circulating Trophoblastic Fetal Cells from Maternal Blood

Thierry_BenjaminBenjamin Thierry, PhD, Professor, Bioengineering, Future Industries Institute, University of South Australia, Australia

Inertial microfluidics has been successfully applied for the enrichment of circulating tumour cells, but its application to the isolation of circulating trophoblastic fetal cells presents additional challenges. We have developed an integrated combining inertial microfluidic and single cell manipulation towards the development of a semi-automated cell-based NIPT compatible with downstream genomic testing.

10:05 Standardized Analysis of Extracellular Vesicles in Liquid Biopsies: From Research to Clinical Applications

Hendrix_AnAn Hendrix, PhD, Assistant Professor, Laboratory of Experimental Cancer Research, Ghent University, The Netherlands

The identification of extracellular vesicle (EV)-associated biomarkers is challenging owing to the complexity of liquid biopsies. We 1) performed quality control studies to identify the impact of (pre-) analytical variables on biomarker identification, 2) developed reference materials to ensure standardized EV measurements, and 3) created EV-TRACK to stimulate researchers to put experimental guidelines into practice. This combined expertise boosted the identification of bacterial EV in the systemic circulation of patients with intestinal barrier dysfunction.

10:35 Multiplex Digital PCR for the Diagnostic of Pilocytic Astrocytoma and Glioneuronal Tumors

Appay_RomainRomain Appay Human Pathologist, Pathological and Neuropathological Anatomy, APHM, Marseille, France

By using Crystal Digital PCR, a multiplex panel was developed for the molecular characterization of pediatric CNS tumors by detecting BRAF; FGFR1 and PIK3C mutations. This permits to get results in 1 day, with high sensitivity and high specificity. This assay can be performed by using DNA with no need of RNA for fusion transcript detection. Thus, dPCR enables laboratories to get access to affordable & fast diagnostic tools.

10:50 Sponsored Presentation (Opportunity Available)

11:05 Coffee Break in the Exhibit Hall with Poster Viewing


PLENARY SESSION

11:35 Moderator’s Remarks

Charlotte Ryckman, Associate, Covington & Burling LLP, Belgium



11:45 Precision Diagnostics in Oncology: Expanding Roles of Liquid Biopsies

Nitzan Rosenfeld, PhD, Senior Group Leader, Cancer Research UK Cambridge Institute, University of Cambridge; CSO, Inivata Ltd., United Kingdom

Effective clinical management relies on accurate diagnostic information, which requires effective techniques and the right samples. Next generation sequencing can provide a wealth of information, but implementing innovative technologies into clinical routine can be a challenge. We’ll examine how analysis of cell-free DNA can provide an opportunity to re-examine many of the current clinical decision points, and a test case for adoption of new diagnostic tools.

12:15 Legal and Regulatory Developments in Precision Medicine and Diagnostic Devices

Erik Vollebregt, Partner, Axon Lawyers, The Netherlands



 

  • What changes will be brought about by the IVDR?
  • What is the impact of the GDPR in the field of precision medicine and diagnostic devices?
  • What are the practical implications of implementation of new European regulations?
  • What are the consequences of the interplay of the IVDR and the GDPR?


12:45
PANEL DISCUSSION: Challenges and Opportunities in European Diagnostic Investments

 

Moderator:

Philippe Peltier, Partner, Kurma Partners, France






Panelists:

Florian Kainzinger, PhD, Managing Partner, Founder, Think.Health Ventures, Germany





Makinen_SeppoSeppo Mäkinen, Partner, Pathena Investments


  • What is different in Europe versus other markets (e.g., US and Israel). How do different European markets compare?
  • What has changed in the landscape of European investments over the past few years? What can be improved?
  • The role of regulators and governments
  • How can start-ups stand out and get attention in the current landscape?

13:30 Close of Enabling Technologies for Circulating Biomarkers