Evidence surrounding the clinical relevance of Circulating Tumour Cells continues to grow. Despite the data, this diagnostic tool has yet to become standard practice in cancer care. At Cambridge Healthtech Institute’s Inaugural Circulating Tumour Cells, leading researchers will demonstrate how they are paving the way to bring liquid biopsy to the patient. Emphasis will be placed on the needs of clinicians, techniques to stratify and monitor, and key applications with the highest probability of success. Tools and techniques for detection, characterization, and isolation will be examined in depth. Finally, CTCs and ctDNA will be evaluated and compared for their use in the clinical management of cancer.

Following Circulating Tumour Cells (April 5-6) will be a dedicated Circulating Cell-Free DNA (April 6-7) conference to continue examining challenges and solutions for circulating biomarkers. Overall, these two events will provide attendees with concrete ideas to ensure success in the clinic.

TUESDAY, 5 APRIL

CTC DETECTION, CHARACTERIZATION, AND ISOLATION

8:00 Registration and Morning Coffee

9:00 Chairperson’s Remarks

Klaus Pantel, M.D., Professor and Founding Director, Institute of Tumor Biology, University Medical Center Hamburg-Eppendorf, Germany

9:35 Detection, Characterization, and ex vivo Expansion of Viable Circulating Tumour Cells

Catherine Alix-Panabières, Ph.D., Director & Associate Professor, Laboratory of Rare Human Circulating Cells, Cell and Tissue Biopathology of Tumors, University Medical Center of Montpellier, France

Circulating tumor cells (CTCs) in blood are promising new biomarkers potentially useful for prognostic prediction and monitoring of therapies in patients with solid tumors. We reported for the first time (1) the establishment of a permanent cell line from CTCs of one colon cancer patient & (2) that PDL1 is heterogeneously expressed on CTCs from metastatic breast cancer patients. CTC research opens a new avenue for understanding the biology of metastasis in cancer patients.

10:05 CTCs Heterogeneity: A Molecular Profiling Workflow with Single-Cell Resolution

Francesca Fontana, Manager, Biology Research & Development, Silicon Biosystems

Despite the several methods for Circulating Tumor Cells (CTCs) enrichment currently available, the CTCs genetic heterogeneity, its relation with tumor stage and dynamics, as well as its clinical significance, still need to be fully investigated. The digital isolation of single pure CTCs enables their genetic characterization within a patient sample, between different patients, or longitudinally across the course of treatment for a given patient, as well as between cells obtained from different enrichment technologies.

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

11:15 Quantification of Malignant Cells in CSF with Cellsearch VERIDEX Improves Diagnosis and Management of Carcinomatous Meningitis

Gilbert C. Faure, Ph.D., PU PH Consultant, Laboratoire d’Immunologie, Université Lorraine & CHU Nancy (Nancytomique), France

Diagnostic methods of leptomeningeal metastases (LM) in Cerebro-Spinal fluid (CSF), lack both specificity and sensitivity. We adapted the Veridex CellSearch® technology to detect Tumour Cells (CSFTCs) in CSF from breast, lung and melanoma cancer patients with LM. This method, established on a limited volume of CSF prove to be of great interest in diagnosis and follow-up of cancer patients with LM opening new fields for characterization of cells crossing the blood-brain-barrier and evaluation of efficiency of systemic or intrathecal therapies.

11:45 A Functionalized Medical Device for CTC Isolation in vivo and Single CTC Molecular Analysis

Shukun Chen, Research Assistant, Institute of Cell Biology, Histology & Embryology, Medical University of Graz, Austria

Using a in vivo isolation technique we currently immunocytochemically characterize CTCs in high-risk prostate cancer patients. However, CTC analysis needs to go beyond mere immunophenotyping which is why we test a new device allowing the recovery of isolated CTCs for the purpose of molecular analysis. Our in vitro study shows that cells captured by the new device, recovered and forwarded to whole genome amplification, array-CGH analysis and next generation sequencing at the single-cell level present with high-quality data, suggesting potential clinical application for personalized medicine.

12:15 CytoQuest™ CR – Circulating Rare Cells Positive Enrichment and Retrieval System

Michaela Adamcova, Research & Diagnostic Products Division, Circulating Tumor Cells Detection, BioVendor - Laboratorni medicina a.s., Czech Republic

Barbora Dvorakova, Research & Diagnostic Products Division, Circulating Tumor Cells Detection, BioVendor - Laboratorni medicina a.s., Czech Republic

CytoQuest™ CR is a non-invasive microfluidic system for capture, enumeration, isolation and retrieval of circulating rare cells (CRCs). A challenge for market adoption of CRCs is the efficient and reproducible identification, single cell isolation, and retrieval of highly pure and viable CRCs.

12:30 Enjoy Lunch on Your Own

EXISTING TECHNOLOGIES AND UNMET NEEDS

14:15 Chairperson’s Remarks

Evi Lianidou, Ph.D., Professor, Analytical Chemistry – Clinical Chemistry, Analysis of Circulating Tumor Cells (ACTC) Lab, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Greece

14:20 Development, Validation, and Clinical Applications of Molecular Assays for the Molecular Characterization of CTCs

Evi Lianidou, Ph.D., Professor, Analytical Chemistry – Clinical Chemistry, Analysis of Circulating Tumor Cells (ACTC) Lab, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Greece

This lecture will be mainly focused on the analytical systems for CTC molecular characterization and its clinical applications in many types of solid cancer. We will also discuss the potential of the molecular characterization of CTC as a liquid biopsy in individualized therapy. This field has a tremendous potential towards the development of molecular assays with potential utility as companion diagnostics, in disease monitoring, or even for early cancer detection.

14:50 3D Microdevice for the in vivo Trapping of Cancer-Associated Circulating Cells

Aline Cerf, Ph.D., CNRS Researcher, NanoBioSystems, LAAS-CNRS, France

We introduce a unique intravascular 3D micro-system for the selective capture of cancer-associated circulating cells directly from the bloodstream. Our methodology is intended to overcome sampling and selection biases of current circulating tumor cell (CTC) detection systems by placing the microdevice in vivo, and by performing CTC trapping based on their physical traits only. Using a fluidic platform reproducing in vivo conditions, we succeeded in capturing PC3 human prostate cancer cells from whole blood in just a few minutes, demonstrating our device’s capability to capture CTCs in conditions close to those found in vivo.

15:20 Role of Epithelial-to-Mesenchymal Transition (EMT) on Circulating Tumor Cell Generation and Metastasis in Prostate Cancer

Alison Allan, Ph.D., Senior Oncology Scientist and Associate Professor, Oncology and Anatomy & Cell Biology, London Regional Cancer Program and Western University, Canada

The U.S. Food and Drug Administration (FDA)-approved CellSearch® system is the current gold standard for CTC enumeration. However, using the CellSearch® approximately 35% of metastatic prostate cancer patients have undetectable CTCs, which may result from the epithelial-to-mesenchymal transition (EMT) and subsequent loss of necessary CTC detection markers. We have developed two pre-clinical assays for assessing human CTCs in xenograft mouse models of metastasis; one that is comparable to the EpCAM-based CellSearch®

15:50 Refreshment Break in the Exhibit Hall with Poster Viewing

CTCs IN PATIENT STRATIFICATION AND THERAPY PREDICTION

16:30 Circulating Tumor Cells (CTC) and Pathological Complete Response (pCR) as Independent Prognostic Factors in Inflammatory Breast Cancer (IBC)

Jean-Yves Pierga, M.D., Ph.D., Circulating Cancer Biomarkers Lab, SiRIC, Translational Research and Medical Oncology, Institut Curie and University Paris Descartes, France

This talk will describe the largest prospective trial in non-metastatic IBC evaluating CTC detection. We observed a high CTC detection rate of 39%, with a strong and independent prognostic value for DFS and OS. Combination of pCR after neoadjuvant treatment, with CTC at baseline, isolates a subgroup of IBC with excellent survival. CTC count should be part of IBC stratification in prospective trials.

17:00 Expansion of Breast Circulating Cancer Cells Predicts Response to Anti-Cancer Therapy

Prashant Kumar, Ph.D., Faculty Scientist, Institute of Bioinformatics, India

This talk will present a new technique that provides an opportunity to analyse CTC clonal heterogeneity and adapt therapeutic modalities in refractory breast cancer patients which may help determine the efficacy of selected therapeutic regimes.

17:30 Clinical Significance of Bone Marrow DTC Detection in Cancer Patients

Nikolay Tupitsyn, M.D., Ph.D., Professor, Oncoimmunology, Haematopoiesis Immunology Lab, Federal State Budgetary Institute, N.N.Blokhin Cancer Research Center, Russia

CTC analysis in cancer patients is now proved to be of great clinical significance. However it cannot fully replace study of bone marrow (BM) DTC. Namely in the bone marrow tumor cells can survive in dormant state for years and decades, then giving rise to distant incurable metastases. We provide data on both detection and clinical significance of DTC in more than 200 patients (breast cancer, ovarian cancer, gastric cancer). While BM is studied excluding hemodilution with the use of modern methods allowing investigation of at least 20 x 106 cells by modern methods, the data received is of great prognostic significance allowing monitoring of treatment protocols efficacy.

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

19:00 Close of Day One

WEDNESDAY, 6 APRIL

CTCs IN PATIENT STRATIFICATION AND THERAPY PREDICTION (Cont.)

8:00 Registration and Morning Coffee

8:40 Chairperson’s Remarks

Julie Lang, M.D., FACS, Associate Professor, Surgery, University of Southern California, United States

8:45 Gene Expression Profiling of Circulating Tumor Cells in Non-Metastatic Breast Cancer

Julie Lang, M.D., FACS, Associate Professor, Surgery, University of Southern California, United States

We hypothesized that transcriptional profiling of CTCs with RNA Seq prior to therapy may predict for pathologic complete response to neoadjuvant chemotherapy in Stage II-III breast cancer. RNA Seq of rare CTCs is feasible in Stage II-III breast cancer and shows evidence of oncogenes and tumor suppressor genes. RNA Seq of CTCs may be performed without background subtraction of leukocytes using our approach. Our preliminary analysis suggests that transcriptional profiling of CTCs may predict for a pathologic complete response to neoadjuvant chemotherapy.

9:15 Molecular Characterization and Genomic Sequencing of Circulating Tumor Cells in Breast Cancer

Aditya Bardia, M.D., Attending Physician, Medical Oncology, Massachusetts General Hospital Cancer Center, United States

Circulating tumor cells (CTCs) can serve as potential “liquid biopsies” offering a potential relatively non-invasive tool for monitoring of breast cancer. We have demonstrated that CTCs can potentially be utilized to monitor response to targeted therapies, to better understand tumor biology, and for the identification of novel actionable targets in breast cancer.

9:45 How Clinical Biobanks Can Support Standardized CTC Assessment to Improve Individualized Medicine: A CTC Guide to Design and Report Trials

Jens K. Habermann, M.D., Ph.D., Professor & Head, Section of Translational Surgical Oncology and Biobanking; Scientific Director, Surgical Center for Translational Oncology-Lübeck (SCTO-L), University of Lübeck & University Medical Center Schleswig-Holstein (UKSH), Germany

Despite current interstudy heterogeneity, current data indicate that CTC detection is of clinical relevance, e.g., as a surrogate prognostic marker in colorectal cancer treatment. This talk will propose a standardized CTC guideline (CTC Guide) to prospectively design and report studies/trials in a harmonized form to overcome interstudy heterogeneity. This will be crucial before implementing CTC detection into clinical consensus guidelines. Hereby, hospital integrated biobanks can play a pivotal role by building the bridge between basic/translational research and clinical routine.

10:15 Coffee Break in the Exhibit Hall with Poster Viewing

Moderator: Aditya Bardia, M.D., Attending Physician, Medical Oncology, Massachusetts General Hospital Cancer Center, United States

• What data/information is needed?
• What are the needs of clinicians?
• How should earlier stage patients be handled?
• How can the process be standardized?

Panelists: Ellen Heitzer, Ph.D., Assistant Professor, Institute of Human Genetics, Medical University Graz, Austria

Evi Lianidou, Ph.D., Professor, Analytical Chemistry – Clinical Chemistry, Analysis of Circulating Tumor Cells (ACTC) Lab, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Greece

Nikolay Tupitsyn, Oncoimmunology, Haematopoiesis Immunology Lab., Federal State Budgetary Institute, N.N.Blokhin Cancer Research Center, Russia

11:45 Close of Conference