Circulating Tumor Cells enable monitoring and diagnosis of disease, however, proof of clinical validity and utility holds this diagnostic tool back from routine use. There is still a need for more predictive and prognostic tools, a clearer understanding of the biology of these cells, and universal standards. Cambridge Healthtech Institute’s Inaugural Circulating Tumor Cells conference will address current challenges and new directions as well as provide solutions and steps forward. Particular focus will be given to addressing tumor heterogeneity and determining relevant cells for treatment decisions. Experts will also discuss the viability of early disease detection along with the need for increased sensitivity and tools to correlate mutations with disease.
TUESDAY, AUGUST 23
7:30 am Main Conference Registration & Morning Coffee
8:30 Chairperson’s Opening Remarks
Joshua M. Lang, M.D., MS, Assistant Professor, Medicine, Carbone Cancer Center, University of Wisconsin
8:40 Predictive and Pharmacodynamic Biomarkers for Prostate Cancer Clinical Trials
Joshua M. Lang, M.D., MS, Assistant Professor, Medicine, Carbone Cancer Center, University of Wisconsin
New technology has been developed for integrated capture and molecular analysis of circulating tumor cells, focused on the androgen receptor signaling pathway. This presentation will discuss technology development, analytic validation and clinical trial results for these new biomarkers in patients with prostate cancer.
9:10 The Role of CTCs in the Development of Predictive Molecular Biomarkers
Tilak K. Sundaresan, Instructor, Medicine, Massachusetts General Hospital Cancer Center
Individual tumor biopsies may provide an incomplete window into the heterogenous nature of acquired drug resistance. It has been suggested that blood-based methods, including the genetic analysis of circulating tumor cells and circulating tumor DNA, may provide a more comprehensive survey of resistance mechanisms to help guide the selection of therapy. Ultimately, however, the clinical utility of blood-based analyses will be determined by their ability to predict clinical responses to drugs. More studies evaluating this premise will be necessary for the field of blood-based diagnostics genetics to fully mature.
9:40 Microfluidic Approaches to CTC Phenotypic Profiling
Shana Kelley, Ph.D., Professor, Biochemistry, University of Toronto
Circulating tumor cells can be highly heterogeneous, and resolving their phenotypic properties is very important to understand their role in cancer progression. We are developing high-throughput microfluidic approaches that can report on CTC phenotypes at the single-cell level. These tools allow dynamic CTC phenotypes to be tracked in patients and animal models of cancer.
10:10 Coffee Break in the Exhibit Hall with Poster Viewing
10:55 Chairperson’s Remarks
Laura Elnitski, Ph.D., Principal Investigator, Translational and Functional Genomics Branch, National Human Genome Research Institute
11:00 Robust Detection of DNA Hypermethylation of ZNF154 as a Pan-Cancer Locus with in silico Modeling for Blood-Based Diagnostic Development
Laura Elnitski, Ph.D., Principal Investigator, Translational and Functional Genomics Branch, National Human Genome Research Institute
Aberrant DNA methylation in cancer represents a potential biomarker for screening and diagnostics. We identified hypermethylation at the ZNF154 CpG island in 15 solid epithelial tumor types from 13 different organs. We further assessed the magnitude and pattern of differential methylation across colon, lung, breast, stomach, and endometrial tumors using next generation bisulfite amplicon sequencing. Our findings strongly support this epigenetic signature as a relevant biomarker for circulating tumor DNA.
11:30 The Analytical Validation of a Blood-Based Biomarker Predictive of Drug Sensitivity for Patients with Prostate Cancer
Daniel Danila, M.D., Genitourinary Oncology Service, Medicine, Memorial Sloan-Kettering Cancer Center
Novel biomarkers hold the promise of stratifying treatments based on the presence of putative targets. Given the effort and cost to develop new biomarkers, it is essential to abide to strict performance metrics when qualifying a promising assay in the context of use.
12:00 pm Development of Reference Materials for the Measurements of Circulating Cell-Free Tumor DNA
Hua-Jun He, Ph.D., Research Biologist, Biosystems and Biomaterials Division, National Institute of Standards and Technology
NIST, in collaboration with the Early Detection Research Network, is developing reference materials for ctDNA. The synthesized plasmid containing 7 gene alteration in cancer was spiked into a background of wild-type genomic DNA at different concentrations. The reference materials were quantified by digital PCR, and will be evaluated by NGS. The ctDNA reference materials will improve the reliability and confidence of the measurement of cell-free DNA biomarkers that show great promise for early cancer detection.
12:30 Enrichment of Circulating Tumor Cells Using a Novel Microfluidic System
Robert Zeillinger, Ph.D., Assoc Professor, Molecular Oncology Group, Medical University of Vienna
The Parsortix microfluidic technology is capable of providing a CTC sample of high purity, which is pre-requisite for applying qPCR to study gene expression levels in CTCs or ddPCR to detect tumor-specific mutations. Based on the enrichment by size and rigidity, the technology is flexible both in regards to the input sample volume and critical step size. We have shown that the efficient reduction of contaminating leukocytes allows the detection of CTC-related transcripts using qPCR at both high sensitivity and specificity.
1:00 Luncheon Presentation: An Automated, Sensitive Microfluidic Device for Capturing and Characterizing CTCs from Whole Blood Samples
Yixin Wang, Ph.D., CSO, Celsee Diagnostics
Current technologies for isolating and analyzing circulating tumor cells (CTCs) are hindered by sub-optimal sensitivity and specificity of devices or assays as well as lack of the capability to characterize CTCs with clinical biomarkers. 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 immunohistochemistry, DNA FISH, RNA FISH, PCR and NGS.
1:30 Refreshment and Cookie Break in the Exhibit Hall with Poster Viewing
2:00 Chairperson’s Remarks
Catherine Alix-Panabières, Ph.D., Director, Laboratory of Rare Human, Circulating Cells (LCCRH), Cellular and Tissular Biopathology of Cancers, University Medical Center of Montpellier
2:05 Establishment and Characterization of a Cell Line from Human Circulating Colon Cancer Cells
Catherine Alix-Panabières, Ph.D., Director, Laboratory of Rare Human Circulating Cells (LCCRH), Cellular and Tissular Biopathology of Cancers, University Medical Center of Montpellier
For the first time, the establishment of a permanent cell line from CTCs of one colon cancer patient has been possible (CTC-MCC-41). In culture for three years, these cells have been characterized at the genome, transcriptome, proteome and secretome levels. CTC-MCC-41 cells resemble characteristics of the original tumor cells in the colon cancer patient and display a stable phenotype characterized by an intermediate epithelial/mesenchymal phenotype, stem-cell like properties and an osteomimetic signature indicating a bone marrow origin.
2:35 Heterogeneity and Dynamics of Circulating Tumor Cells
Anders Carlsson, Ph.D., David and Dana Dornsife, College of Letters, Arts and Sciences, University of Southern California
The HD-SCA assay is capable of identifying and characterizing rare events, like CTCs, in blood and bone marrow aspirates of cancer patients, using an enrichment-free, “no cells left behind” imaging based approach. Subsequent next generation single cell sequencing of candidate cells has been applied longitudinally in several cancer diseases, revealing the dynamics of tumor cell clonality at different disease stages and treatments.
3:05 Dissecting CTC Phenotypes in Mechanisms of Breast Cancer Dormancy
Dario Marchetti, Ph.D., Director, Center of the Biomarker Research Program, The Methodist Hospital Research Institute
In this talk, we will discuss the isolation and characterization of CTC subsets from peripheral blood of patients diagnosed with or without breast cancer brain metastasis (BCBM). We identified proliferative and invasive properties of 3D CTC tumorspheres distinctive upon uPAR/int β1 combinatorial expression. The molecular characterization of uPAR/int β1 CTC subsets will enhance abilities to prospectively identify patients who may be at high risk of developing BCBM.
3:35 Overcoming the Challenges of Heterogeneous Liquid Biopsies through Precise Image-Based Single-Cell Sorting
Farideh Bischoff, Ph.D.; Executive Director, Scientific Affairs; Head, Clinical Development Lab, Menarini Silicon Biosystems
Heterogeneity has always limited the effectiveness of molecular analysis for solid tumors, including circulating tumor cells that are derived from these heterogeneous solid tumors. DEPArrayTM is an innovative technology capable of sorting and isolating 100% pure single or pooled cells through a digitally controlled Dielectrophoretic field using a semiconductor chip. A complete workflow for recovery of single or pools of pure circulating tumor cells from enriched blood has been established and will be presented. Furthermore, studies demonstrating utility of this workflow for downstream analysis using NGS will be reviewed.
4:05 Refreshment Break in the Exhibit Hall with Poster Viewing
4:50 PANEL DISCUSSION: Circulating Biomarkers for Early Detection: Promise and Reality
Moderator:Stuart S. Martin, Ph.D., Associate Professor, Physiology, Greenebaum Cancer Center, University of Maryland School of Medicine
Panelists:
Klaus Pantel, M.D., Professor and Founding Director, Institute of Tumor Biology, University Medical Center Hamburg-Eppendorf, University of Hamburg
Catherine Alix-Panabières, Ph.D., Director, Laboratory of Rare Human, Circulating Cells (LCCRH), Cellular and Tissular Biopathology of Cancers, University Medical Center of Montpellier
Joshua M. Lang, M.D., MS, Assistant Professor, Medicine, Carbone Cancer Center, University of Wisconsin
Laura Elnitski, Ph.D., Principal Investigator, Translational and Functional Genomics Branch, National Human Genome Research Institute
- Challenges in getting to early detection?
- Detecting low levels of disease
- Cutting down on false positive testing?
- Correlating results with disease
5:50 Wine & Cheese Pairing Welcome Reception in the Exhibit Hall with Poster Viewing
6:00 Dinner Short Course Registration
6:15 Close of Day
WEDNESDAY, AUGUST 24
7:15 am Registration
7:30 Problem-Solving Breakout Discussions with Continental Breakfast
Understanding CTC Heterogeneity
Anders Carlsson, Ph.D., David and Dana Dornsife, College of Letters, Arts and Sciences, University of Southern California
- What do we mean with heterogeneity? Variation around one vs. several 'mean values' of a given property?
- What are the important variables where heterogeneity exist in CTCs?
- Is heterogeneity a cell population 'property' or a mix of cell populations'?
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8:25 Chairperson’s Remarks
Andrew D. Rhim, M.D., Assistant Professor, Internal Medicine, Division of, Gastroenterology, University of Michigan
8:30 Novel Methods for Eliminating Wild-Type Alleles from Liquid Biopsies
G. Mike Makrigiorgos, Ph.D., Professor, Radiation Oncology, Dana-Farber and Harvard Medical School
We present Nuclease-assisted Mutation Enrichment with Overlapping Probes (NaME-PrO), a simple and powerful technology for eliminating wild-type DNA sequences from CTCs and circulating DNA, in order to focus on clinically relevant DNA alterations. Furthermore, we show that NaME-PrO combines seamlessly with existing technologies such as COLD-PCR, digital PCR, high resolution melting or sequencing.
9:00 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.
9:30 Isolation and Characterization of Biologically Relevant Circulating Tumors Cells (CTCs): The Importance of the CTC Microenvironment
John J. O’Leary, M.D., Ph.D., MSc, MA, FRCPath, FFPathRCPI, 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)
Circulating tumor cells (CTCs) offer a unique biological window on cancer cell metastasis. Sets of unique biological interactions visited upon the CTC produce a pro-survival and pro-invasion phenotype. CTCs demonstrate different physical form states and different biological transitions during the metastatic trafficking process. This talk addresses issues central to CTC physical state analysis, developing CTC molecular classifiers of biological relevance and critically evaluates the microenvironment of CTCs from primary to liquid phase and pre-metastatic niches, in order to fully define those CTCs capable of forming metastases.
10:00 Liquid Biopsy in Clinical Practice: Implementation of the OncoBEAM RAS Platform in the Management of mCRC Patients
Vishal Sikri, Vice President, Commercial Operations, Sysmex Inostics
Measurement of ctDNA is being widely studied across different cancer types. The use of ctDNA assays will continue moving forward since they allow for high sensitivity while still maintaining specificity. The OncoBEAMTM platform has been published across cancer types because of its high sensitivity compared to other technologies. The OncoBEAMTM RAS IVD assay was launched in 2016 for therapy selection and resistance detection applications highlighting the current and future value of ctDNA testing in oncology.
10:30 Coffee Break in the Exhibit Hall with Poster Viewing
12:40 pm Luncheon Presentation: Sensitive Assays to Detect Mutant ESR1 in Plasma Circulating Tumor DNA from Breast Cancer Patient Samples
Gaorav Gupta, Ph.D., Assistant Professor, University of North Carolina at Chapel Hill
Mutant estrogen receptor (ESR1) emerged as a mechanism for metastatic breast cancer (MBC) endocrine therapy resistance. ESR1 mutations are rarely identified in untreated primary breast cancers, but are present in ≤ 30% of patients with ER+ MBC. The most common mutations are in the ESR1 ligand-binding domain and could confer resistance. We describe a multiplexed dPCR ctDNA assay, detected allelic burden changes with longitudinal sampling, and developed a ThunderBolts assay to screen for additional mutations.
1:20 Close of Circulating Tumor Cells
1:25 Ice Cream and Cookie Break in the Exhibit Hall with Poster Viewing