Many technologies have been developed for the application of circulating tumor cells, including techniques for isolation, enumeration, characterization, propagation and genomic profiling. However, as yet there is only one approved technology for clinical use. This year’s Sixth Annual Circulating Tumor Cells conference will delve into new insights in the field of circulating tumor cell research and clinical application by investigating key challenges for advancement into the clinic. The visionary faculty will look at comparisons of technical progress and advancement for a range of circulating biomarkers including cell-free DNA, exosomes and extracellular vesicles and illuminate strategies for moving them into clinical use.
Scientific Advisory Board
Stefanie Jeffrey, M.D., John and Marva Warnock Professor, Surgery; Chief, Surgical Oncology Research, Stanford University School of Medicine
Stuart S. Martin, Ph.D., Associate Professor, Physiology, Greenebaum NCI Cancer Center, University of Maryland School of Medicine
Klaus Pantel, M.D., Professor and Founding Director, Institute of Tumor Biology, University, Medical Center Hamburg-Eppendorf
Steven A. Soper, Ph.D., Professor, Biomedical Engineering & Chemistry; Associate Editor, Analyst; Director, Center for BioModular Multiscale Systems, University of North Carolina
Monday, March 7
10:30 am Conference Program Registration Open
11:50 Chairperson’s Opening Remarks
Klaus Pantel, M.D., Professor and Founding Director, Institute of Tumor Biology, University Medical Center Hamburg-Eppendorf
12:00 pm Liquid Biopsy: Expectations and Required Steps to Bring It into Clinical Practice
Klaus Pantel, M.D., Professor and Founding Director, Institute of Tumor Biology, University Medical Center Hamburg-Eppendorf
Circulating tumor cells (CTCs), nucleic acids (ctDNA, cfmiRNA) and exosomes in the blood of cancer patients have received increasing attention as new diagnostic tool enabling “liquid biopsies”. The perspective to avoid invasive tissue biopsies and obtain similar or even more information by a “simple” blood test has enormous implications in cancer diagnostics. Here the expectations and future steps required to bring liquid biopsies into clinical practice will be discussed.
12:30 Circulating Tumor Cells as a Biomarker of Treatment Efficacy and Prediction in Castration-Resistant Prostate Cancer
Howard I. Scher, M.D., Chief, Genitourinary Oncology Service; Member and Attending Physician, Department of Medicine, Memorial Sloan Kettering Cancer Center; Professor, Medicine, Weill Cornell Medical College
Pre and post-therapy circulating tumor number has been shown to be prognostic for survival. Use in clinical practice and in drug development as a measure of treatment efficacy has been more limited. Analyses of the strength of association of the enumeration biomarker with survival will be discussed along with a recent analysis showing that a CTC enumeration biomarker in combination with LDH was shown to meet the Prentice Criteria for individual-patient level surrogacy. Use of a predictive biomarker to guide treatment selection will also be discussed.
1:00 Session Break
1:15 Luncheon Presentation I: Size-Based Isolation of
Circulating Tumor Cells Associated to Droplet Digital PCR
Allow Prediction of KRAS Mutations in Patients with
Colorectal Cancer before Tumor Surgery
Jérôme A. Denis, M.D., Ph.D., Sorbonne Universités
1:45 Luncheon Presentation II: Comprehensive
Ultra-Sensitive CTC Analysis in the Management of Cancer
Veena Singh, M.D.,Senior Vice President & Senior Medical Director, Biocept, Inc.
Circulating tumor cells (CTCs) isolated solely by EPCAM capture and their CK positive, CD45 negative profile miss CK negative non-EPCAM expressing CTCs. These maybe cancer stem cells or those with epithelial mesenchymal transition, and possibly more clinically relevant. Hence we developed a multi-antibody approach for characterization of all CTC phenotypes.
2:15 Session Break
2:30 Chairperson’s Remarks
Stefanie Jeffrey, M.D., John and Marva Warnock Professor, Surgery; Chief, Surgical Oncology Research, Stanford University School of Medicine
2:40 FEATURED PRESENTATION: Circulating Epithelial Cells as Biopsies of Subclinical Cancer: Novel Technologies for Molecular Analysis
Andrew D. Rhim, M.D., Assistant Professor, Internal Medicine, Division of Gastroenterology, University of Michigan
Our studies in genetic mouse models have shown that dysplastic epithelial cells are shed into the blood stream long before the formation of clinically detectable tumors. Here, we will summarize our efforts in utilizing circulating epithelial cells as a sampling of clinically undetectable dysplastic lesions that may represent the earliest forms of cancer. This discussion will include recent work in the use of novel microfluidic platforms to achieve ultra-sensitive genomic analysis of captured cells by any platform.
3:10 Disease Associated Immune Cells as Novel Biomarkers for Liquid Biopsy
Lidia C. Sambucetti, Ph.D., Senior Program Director, Cancer Research Technologies, Biosciences Division, SRI International
Liquid biopsies enable the collection of information on a cancer patient’s disease from a blood test based on enumeration of circulating tumor cells (CTCs) and characterization of their protein or genetic biomarkers. Using our ultra-high throughput non-enrichment rare cell detection platform, FASTcellTM, we now extend the use of liquid biopsies to the detection of rare biomarkers on immune cells for detection of features of the tumor microenvironment and for certain infectious diseases.
3:40 Platelet Cloaking, Cancer Cells and CTCs: Lessons for the Metastatic Cascade
John O’Leary, M.D., Ph.D., Chair, Pathology, Trinity College Dublin; Director, Pathology, Coombe Women and Infants University Hospital; Consultant, Pathologist, St. James’s Hospital; Principal Investigator, Biomedical Diagnostics Institute (BDI)
The talk will critically examine the biological pathways involved in these interactions and will describe novel immune surveillance inhibition mechanisms adopted by platelet cloaked cancer cells directly inhibiting natural killer [NK] cell function, including direct receptor-ligand perturbation and immune decoy defense mechanisms.
4:10 Recovery of Single CTCs Using DEPArray™ for Precise Downstream Molecular Profiling and NGS Applications
Farideh Bischoff, Ph.D., Executive Director, Scientific Affairs & Head of Clinical Development, Silicon Biosystems
DEPArray™ allows recovery of pure and intact target (rare) cells. Given heterogeneity among CTCs , molecular analysis of individual and intact cells is critical to ensure that low level genomic alterations are observed. A workflow for DEPArray-based recovery of individual CTCs is now established.
4:25 Going Beyond Averages and Enumeration – How Epic’s
‘No Cell Left Behind’ Platform is Driving Novel Insights in Cancer
Murali Prahalad, Ph.D., President & CEO, Epic Sciences
Cancer is a disease characterized by incredible cellular heterogeneity often driven by multiple clonal species possessing unique proteomic and genomic profiles. This session will highlight how Epic Sciences’ unique enrichment-free approach enables new insights into cancer that surpass simple enumeration or the genomic averaging that results from the sequencing of tissue biopsy or cfDNA.
4:40 Refreshment Break and Transition to Plenary Session
5:00 Plenary Keynote Session
6:00 Grand Opening Reception in the Exhibit Hall with Poster Viewing
7:30 Close of Day
Tuesday, March 8
7:00 am Registration Open and Morning Coffee
8:00 Plenary Keynote Session
9:00 Refreshment Break in the Exhibit Hall with Poster Viewing
10:05 Chairperson’s Remarks
Klaus Pantel, M.D., Professor and Founding Director, Institute of Tumor Biology, University Medical Center Hamburg-Eppendorf
10:15 Detection, Characterization and ex vivo Expansion of Viable Circulating Tumor Cells
Catherine Alix-Panabières, Ph.D., Maître de Conférence - Praticien Hospitalier, Associate Professor, Director, Laboratory of Rare Human Circulating Cells, Institute of University Clinical Research (IURC), University Medical Centre of Montpellier, University of Montpellier
Circulating Tumor Cells (CTCs) in blood are promising new biomarkers potentially useful for prognostic prediction and monitoring of therapies in patients with solid tumors including colon cancer. However, an in-depth investigation of CTCs is hampered by the very low number of these cells, especially in the blood of colorectal cancer patients. Here we describe the establishment of cell cultures and a permanent cell line from CTCs of one colon cancer patient.
10:45 Detection and Molecular Profiling of Single CTCs and a Patient Derived CTC Line
Tobias M. Gorges, Ph.D., Senior Scientist, Department of Tumor Biology, Center of Experimental Medicine, University Medical Center Hamburg-Eppendorf
Detection and molecular characterization of circulating tumor cells (CTCs) is challenging. We compared different approaches for CTC enrichment. A reliable protocol for the molecular characterization of single cells - targeting up to 84 transcripts per single cell - could be established. We also could establish and characterize a patient derived CTC line. Our findings help to understand the complexity of tumor biology.
11:15 A Direct, Chip-Based Approach to Analysis of Cell-Free Nucleic Acids
Shana Kelley, Ph.D., Professor, Biochemistry, University of Toronto
The analysis of cell-free nucleic acids (cfNA) can reveal the mutational spectrum of a tumor without the need for invasive sampling of tissue; however, this requires the differentiation of nucleic acids that originate from healthy cells from the mutated sequences shed by tumor cells. Here we report an electrochemical clamp assay that directly detects mutated sequences in patient serum. This is the first successful detection of cfNA without the need for enzymatic amplification, a step that normally requires extensive sample processing.
11:45 Isolation and ex vivo Expansion of CTCs for Molecular Diagnosis and Drug Testing
Sunitha Nagrath, Ph.D., Assistant Professor, Chemical Engineering, University of Michigan
Currently, tumor genotyping is performed from tumor biopsies or re-biopsies. CTCs may provide a non-invasive alternative to traditional biopsy to identify the key genetic signatures and resistance mechanisms. There is currently no way to predict which of the many early phase compounds that target resistance will lead to further response. Here we present a strategy to culture CTCs ex vivo reproducibly and allow us to perform ex vivo drug testing.
12:15 pm Session Break
12:25 Luncheon Presentation I: Gene Expression Profiling
of Circulating Tumor Cells in Breast Cancer
Julie E. Lang, M.D., FACS, Associate Professor, Breast and Soft Tissue Surgery,
Norris Comprehensive Cancer Center, University of Southern California
CTCs are prognostic in all stages of breast cancer, yet few studies have examined their role as predictive biomarkers. Affinity based and microfluidic chip-based technologies may be used for sequencing the RNA of CTCs. The current state and what is necessary to move CTCs from a research grade biomarker to a clinical grade biomarker will be discussed.
12:55 Luncheon Presentation II: An Automated, Sensitive Microfluidic Device for Capturing and Characterizing CTCs from Whole Blood Samples
Yixin Wang, Ph.D., CSO, 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.
1:25 Refreshment Break in the Exhibit Hall with Poster Viewing
2:00 Chairperson’s Remarks
Steven A. Soper, Ph.D., Professor, Biomedical Engineering & Chemistry; Associate Editor, Analyst; Director, Center for BioModular Multiscale Systems, University of North Carolina
2:10 Circulating Tumor Cells and Mouse Models of Serous Ovarian Cancer
Victoria Bae-Jump, M.D., Ph.D., Associate Professor, Division of Gynecologic Oncology, University of North Carolina
Ovarian cancer (OC) screening is considered the “holy grail” of gynecologic oncology, given that attempts using CA-125 and other tumor markers as well as ultrasound imaging have largely been unsuccessful in screening for this disease. Our novel circulating tumor cell assay employs a dual selection strategy and is being explored using a genetically engineered mouse model (K18-gT121+/-;p53fl/fl;Brca1fl/fl) of OC, providing access to well-defined early and late stage disease blood samples.
2:40 Orthotopic Mouse Models of Cancer and GFP Labeling for the Study of Circulating Tumor Cells
Robert M. Hoffman, Ph.D., President, AntiCancer, Inc.; Professor, Surgery, University of California, San Diego
We have previously determined that orthotopic mouse models of cancer produce viable circulating tumor cells (CTCs) in contrast to ectopic models. We have also demonstrated that green fluorescent protein (GFP) labels CTCs for further analysis such as metastatic potential or chemosensitivity testing. Patient CTCs can also be selectively labeled with GFP ex vivo for analysis.
3:10 Early Pancreatic Markers, Biomarkers for Exosome Isolation
Raghu Kalluri, M.D., Ph.D., Professor & Chair, Cancer Biology, University of Texas MD Anderson Cancer Center
This talk will review our work with exosomes and how we’ve used mass spectrometry analyses to identify a cell surface proteoglycan, glypican-1 (GPC1), specifically enriched on cancer-cell derived exosomes. We’ll discuss how we monitored and isolated these by using flow cytometry from the serum of patients and mice with cancer. We will discuss how GPC1(+) crExos may serve as a potential non-invasive diagnostic and screening tool to detect early stages of pancreatic cancer to facilitate possible curative surgical therapy.
3:40 Liquid Biopsy Using Exosome RNA and ctDNA, Realizing the “Holy Grail” of Personalized Medicine
Johan Skog, Ph.D., CSO, Research & Development, Exosome Diagnostics
Recently, ctDNA has shown promise for cancer mutation detection. ExoDx platform of exoRNA + ctDNA addresses the limitations of ctDNA alone. ExoRNA is abundant from bio-fluids and yields mutations, splice variants, and fusions.
4:10 St. Patrick’s Day Celebration in the Exhibit Hall with Poster Viewing
5:00 Breakout Discussions in the Exhibit Hall
These interactive discussion groups are open to all attendees, speakers, sponsors, & exhibitors. Participants choose a specific breakout discussion group to join. Each group has a moderator to ensure focused discussions around key issues within the topic. This format allows participants to meet potential collaborators, share examples from their work, vet ideas with peers, and be part of a group problem-solving endeavor. The discussions provide an informal exchange of ideas and are not meant to be a corporate or specific product discussion.
Patient-Derived Orthotopic Xenografts
Robert M. Hoffman, Ph.D., President, AntiCancer, Inc.; Professor, Surgery, University of California, San Diego
- Patient-derived orthotopic xenografts (PDOX) models
- Better mimic of metastasis than subcutaneous xenografts
- Orthotopic tumor mouse models can replicate patient tumor behavior
- Subcutaneous tumor mouse models do not replicate patient tumor behavior
- Patient-derived orthotopic xenograft models have important potential for individualized cancer therapy
Challenges and Opportunities for Therapeutically Targeting Cells in Blood
Michael R. King, Ph.D., Daljit S. and Elaine Sarkaria Professor of Biomedical Engineering, Meinig School of Biomedical Engineering, Cornell University
- Unique aspects of the vascular microenvironment (shear stress, blood cells and plasma)
- The “best” and most relevant animal models of metastatic cancer
- Clinical translation and next steps
6:00 Close of Day
Wednesday, March 9
7:00 am Registration Open
7:00 Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee
8:00 Plenary Keynote Session Panel
10:00 Refreshment Break and Poster Competition Winner Announced in the Exhibit Hall
10:50 Chairperson’s Remarks
Steven A. Soper, Ph.D., Professor, Biomedical Engineering & Chemistry; Associate Editor, Analyst; Director, Center for BioModular Multiscale Systems, University of North Carolina
11:00 KEYNOTE PRESENTATION: New Tools for the Isolation of Circulating Markers: Integrated Microfluidic Systems for the Efficient Isolation of CTCs, Cell-Free DNA and Exosomes
Steven A. Soper, Ph.D., Professor, Biomedical Engineering & Chemistry; Associate Editor,
Analyst; Director, Center for BioModular Multiscale Systems, University of North Carolina
Liquid biopsies are generating interest within the biomedical community due to
the simplicity for securing important markers to realize precision medicine. These
circulating markers consist of whole cells such as CTCs, molecules such as cell-free
DNA and nanovesicles such as exosomes. We are developing a microfluidic system
that can process whole blood and select all three of these markers from a single
blood sample.
11:30 Why Do Cancer Cells Move?
Daniel Irimia, M.D., Ph.D., Assistant Professor, Division of Surgery, Science & Bioengineering, Massachusetts General Hospital and Harvard Medical School; Associate Director, BioMEMS Resource Center
When cancer cells leave the tumor, they set in motion a cascade of events that ends with the formation of distant metastases and the death of 90% of cancer patients. However, despite their importance, our understanding of the conditions that trigger cancer cell migration is very limited. In this presentation, I will discuss the design of novel microfluidic tools which revealed the unexpected ability of cancer cells to navigate microscopic mazes along the shortest path and helped identify novel mechanisms for the cancer cell migration.
12:00 pm Extracellular Vesicles as Couriers of Cancer Information
Hakho Lee, Ph.D., Assistant Professor, Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School
Tumor cells release a variety of extracellular vesicles (EVs) which carry information in the form of protein, DNA and RNA. For example, microRNAs can be transferred via EVs to normal cells, thereby changing their phenotype in support of tumor progression. These vesicles and other extracellular RNA vehicles also carry a representation of the transcriptome of the tumor cells which can be found in biofluids and reports on the genetic drivers and status of the cancer.
12:30 Session Break
12:40 Luncheon Presentation: Telomerase-directed tagging of Circulating Tumor Cells
Jay F. Dorsey, M.D., Ph.D., Assistant Professor, Department of Radiation Oncology, University of Pennsylvania
In this presentation we will discuss the association of “telomerase-dependent CTC findings” with treatment response in a variety of cancer patient populations including malignant glioma and non-small cell lung cancer (NSCLC) along with an analysis of molecular driver characterization in CTCs.
1:10 Refreshment Break in the Exhibit Hall and Last Chance for Poster Viewing
1:50 Chairperson’s Remarks
Hsian-Rong Tseng, Ph.D., Professor, Molecular & Medical Pharmacology; Member, JCCC Cancer Molecular Imaging; Faculty, Crump Institute for Molecular Imaging, University of California, Los Angeles
2:00 The NCI SBIR/STTR Program: Accelerating the Commercialization of New CTC Technologies
Xing-Jian Lou, Ph.D, Program Director, SBIR Development Center, National Cancer Institute
The federal SBIR/STTR program is one of the largest sources of early-stage technology financing in the U.S. Under this program, the National Cancer Institute (NCI) spends $120 million annually to support R&D and commercialization of novel technologies for cancer prevention, diagnostics and treatment, including CTC technologies. In this presentation, Dr. Xing-Jian Lou will discuss efforts to help NCI-funded small businesses overcome regulatory hurdles in the translation of new technologies.
2:15 PANEL DISCUSSION
Steven A. Soper, Ph.D., Professor, Biomedical Engineering & Chemistry; Associate Editor, Analyst; Director, Center for BioModular Multiscale Systems, University of North Carolina
Murali Prahalad, Ph.D., President and CEO, Epic Sciences
Farideh Bischoff, Ph.D., Executive Director, Scientific Affairs, Silicon Biosystems, Inc.
Veena Singh, M.D., Senior Vice President & Senior Medical Director, Biocept, Inc.
3:30 Analyzing Liquid Biopsy Samples with Next-Generation Sequencing in Clinical Research
4:00 Session Break
4:10 Chairperson’s Remarks
Stuart S. Martin, Ph.D., Associate Professor, Physiology, Greenebaum NCI Cancer Center, University of Maryland School of Medicine
4:15 Small Cell Lung Cancer Circulating Tumor Cell-Derived Explants – A New Preclinical Platform for Examining Drug Response
Kristopher Frese, Ph.D., Associate Scientist, Cancer Research UK, Manchester Institute, The University Of Manchester
Due in part to the paucity of tissue samples available for preclinical studies, the past 30 years of research have not provided any new therapeutic options for small cell lung cancer (SCLC). We have recently demonstrated that SCLC circulating tumor cells (CTCs) are tumorigenic in mice and the resulting patient CTC-derived explants (CDX) mirror patients’ disease both in vivo and in vitro. Studies examining the therapeutic efficacy of novel targeted agents are currently underway.
4:45 Antibody-Free Capture of Tumor Cells from Blood and Novel CTC-Targeting Therapeutics
Michael R. King, Ph.D., Daljit S. and Elaine Sarkaria Professor of Biomedical Engineering, Meinig School of Biomedical Engineering, Cornell University
Our laboratory uses surfactant-nanotube complexes to enhance E-selectin-mediated capture and isolation of tumor cells without the use of capture antibodies. Functionalization with sodium dodecanoate surfactant induces a switch to firm cellular adhesion of tumor cells with a simultaneous de-adhesion of blood leukocytes. This system has proved valuable in testing new liposome-based therapeutics designed to target tumor cells in blood for the prevention of metastasis.
5:15 POSTER 99: Microfluidic Isolation of Circulating Tumor Cells from Different Venous Sources in Early Lung Cancer
Vasudha Murlidhar, Ph.D., Candidate, Nagrath Lab, Department of Chemical Engineering, University of Michigan, Ann Arbor
5:45 Close of Conference Program